MACROCYCLIC GLUCAGON-LIKE PEPTIDE 1 RECEPTOR AGONISTS

AGONISTES MACROCYCLIQUES DU RECEPTEUR DU PEPTIDE 1 DE TYPE GLUCAGON

English Abstract

In an embodiment, the present invention provides a compound of the formula: or a pharmaceutically acceptable salt thereof, and methods of using this compound for treating type II diabetes mellitus.

French Abstract

Dans un mode de réalisation, la présente invention concerne un composé de formule : ou un sel pharmaceutiquement acceptable de celui-ci, et des méthodes d'utilisation de ce composé pour le traitement du diabète sucré de type II.

Claims

-338-
Claims
1. A compound of the formula:
A
r R4
Y7
s___ Z3
B Y3 I õ2 N i _R5
I Y 1 y4 2 z Z , y6
Y5-
wherein -A- is -CRaRbCRaRbCRbRb0-, -OCRbRbCRaRbCRaRb-, -OCRbRbCRbRb0-,
-CRaRbCRbRbOCRbRb-, -CRbRbOCRbRbCRaRb-, -CRbRbOCRbRb-,
-CRaRbCRbRb0- or -OCRbRbCRaRb-;
W at each occurrence is independentlyfl, halo, Ci-C2alkyl, OH or Ci-C3a1koxy;
Rb at each occurrence is independently H, halo or Ci-C2alkyl;
a
_1
2 A a X6 -5x5 x-----
.)( b ----x b
is or , wherein a is the point of
attachment to linker A;
b is the point of attachment of linker B;
X1, X2, X3 and X4 are independently N, CH or CR1, wherein no more than two of
X1, X2,
X3 and X4 are N and no more than two of X1, X2, X3 and X4 are CR1;
X5 is N, CH or CR1a, X6, X7 and X8 are independently N, CH or CR1, wherein no
more
than two of X5, X6, X7 and X8 are N and no more than two of X5, X6, X7 and X8
are CR"
or CR1;
R.1 at each occurrence is independently CN; halo; Cl-C3alkyl optionally
substituted with
01-1; Ci-C3haloalkyl; Ci-C3alkoxy; C3-05cycloalkyl; -SO2Ci-C3alkyl;
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o Rh
Re y9 /
9 Re )cx9
Re'
N 9 X9 N
.\N '")(' X
Re or
wherein each X9 is independently CH
or N and no more than one X9 in the ring is N, each Re is independently
selected from: H,
Ci-C3haloalkyl, halo, C3-05cycloalkyl and C1-C3alkyl optionally substituted
with OH, Rh
is H, Ci-C3haloalkyl, halo, C3-05cycloalkyl, OH, -NRcRd or Ci-C3alkyl
optionally
substituted with OH;
5- or 6- membered heteroaryl or phenyl wherein the heteroaryl or phenyl is
optionally
substituted with one or two substituents independently selected from: Ci-
C3a1koxy, C3-
C5cycloalkyl, -C112-C3-05cycloalkyl, -S02C1-C3alkyl, C4-05heterocyclyl, -CH2-
C4-
05heterocyclyl, halo, Ci-C3haloalkyl, Ci-C3haloalkoxy, CN, -CONRcRd, -NRcR" or
Ci-
C3alkyl optionally substituted with OH;
Rla is CN; halo; Ci-C3alkyl optionally substituted with OH; Ci-C3haloalkyl; or
Ci-
C3a1koxy;
-B- is -CH20-, -OCH2- or -CH2NH-;
Y1 , y2 and Y7 are independently N, CH or CR2, wherein no more than one of Y1,
Y2 and
y7 is N and no more than two of y1, y2 and y7 is CR2;
y3, y4, Y5 and Y6 are independently N, CH or CR2, wherein no more than two of
Y3, Y4,
Y5 and Y6 are N and no more than two of Y3, Y4, Y5 and Y6 are CR2;
R2 at each occurrence is independently halo or methyl;
Z1, Z2 and Z3 are independently N, CH or CR3, wherein no more than two of Z1,
Z2 and
Z3 are N and no more than two ofzi, Z2 and Z3 are CR3;
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R3 at each occurrence is independently halo; C1-C4alkyl; -0C4-C6cycloalkyl
optionally
substituted with C1-C2alkoxy, OH, Ci-C3alkyl or C1-C3haloalkyl; -0C4-
C6heterocycly1
optionally substituted with C1-C2a1koxy, OH, C1-C3alkyl or C1-C3haloalkyl; or
Ci-
C4a1koxy optionally substituted with one or two substituents selected from: C1-
C2alkoxy,
OH, -NRfRg, -CONRcRd, CN, halo or 5- or 6- membered heteroaryl optionally
substituted
with C1-C3alkyl;
N _______________________________________ A
0
R4 is orNN
=
o
__________________________________________ 9\ /0
N-S
N-- H
R5 is -CO2H, Or 5
1 0 Rc and Rd are each independently H or C1-C3alkyl;
Rf is H or C1-C3alkyl; and
Rg is H, C1-C3alkyl, C1-C3haloalkyl, C3-05cycloalkyl, C(0)C1-C3alkyl, or C1-
C3a1ky1C3-
Cscycloalkyl;
or a pharmaceutically acceptable salt thereof.
2. The compound according to claim 1, wherein the compound
is of the formula:
A
Y7 N 3
,Y3 I y2 VR5
1 2
y4 ys
Y5'
or a pharmaceutically acceptable salt thereof.
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3. The compound according to claim 1 or claim 2, wherein
2 X1 a
I 3 I
x
b
is , or a pharrnaceutically acceptable salt
thereof.
4. The compound according to claim 3, wherein X1, X3 and X4 are CH, X2 is
CR1
H3CO2S
,
I-1 N
N
and R1 is CN, Cl, F, CF3,
O
N'
õNõ,
?C, = or , or a pharmaceutically acceptable
salt thereof.
5. The compound according to claim 3, wherein X1 is N; X2 is C(CF3); and X3
and
X4 are CH, or a pharmaceutically acceptable salt thereof.
6. The compound according to claim 3, wherein X1, X3 and X4 are CH; and X2
is N,
or a pharmaceutically acceptable salt thereof.
7. The compound according to claim 3, wherein V and X4 are CH; X2 is
C(CF3);
and X3 is N, or a pharmaceutically acceptable salt thereof.
8. The compound according to claim 3, wherein X' and X3 are CH; X2 is
C(CN); and
X4 is N, or a pharmaceutically acceptable salt thereof.
9. The compound according to claim 3, wherein X1 and X3 are CH; X2 and X4
are
CR1; and each R1 is independently selected from F, CI and CN, or a
pharmaceutically
acceptable salt thereof.
10. The compound according to claim 1 or claim 2, wherein
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a
Xs -X5
is , or a pharmaceutically acceptable salt
thereof
11. The compound according to claim 10, wherein X5, X7 and X8 are CH and X6
is
C(CN), or a pharmaceutically acceptable salt thereof.
12. The compound according to claim 10, wherein X5 is N; X6 is C(CN); and
X7 and
X8 are CH, or a pharmaceutically acceptable salt thereof
13. The compound according to any one of claims 1 to 12, wherein -A- is -
CH2CH2CH20-, -CH2OCH2-, -CH2CH2OCH2-, CH2OCH2CH2- Or -CF2CH2OCH2-, Or a
pharmaceutically acceptable salt thereof.
14. The compound according to any one of claims 1 to 13, wherein -B- is -
CH20-, or
a pharmaceutically acceptable salt thereof
15. The compound according to any one of claims 1 to 14, wherein Z1 is CH
or CR3;
and R3 is F, -OCH3, -OCH2CH2OCH3, OCH2CH2OH or OCH2CH2N(CH3)2, or a
pharmaceutically acceptable salt thereof.
16. The compound according to any one of claims 1 to 15, wherein R5 is -
CO2H, or a
pharmaceutically acceptable salt thereof.
17. A pharmaceutical composition comprising a compound, or a
pharmaceutically
acceptable salt thereof, according to any one of claims 1 to 16 and at least
one
pharmaceutically acceptable carrier, diluent, or excipient.
18. A compound, or a pharmaceutically acceptable salt thereof, according to
any one
of clainls 1 to 16 for use in therapy.
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19. A compound, or a pharmaceutically acceptable salt thereof, according to
any one
of claims 1 to 16 for use in the treatment of type H diabetes mellitus.
20. A compound, or a pharmaceutically acceptable salt thereof, according to
any one
of claims 1 to 16 for use in lowering blood glucose levels.
21. A compound, or a pharmaceutically acceptable salt thereof, according to
any one
of clairns 1 to 16 for use in the treatment of hyperglycemia.
22. A compound, or a pharmaceutically acceptable salt thereof, for use
according to
any one of claims 18 to 21, wherein the compound is administered orally.
23. Use of a compound, or a pharmaceutically acceptable salt thereof,
according to
any one of claims 1 to 16 in the manufacture of a rnedicament for the
treatment of type II
diabetes mellitus.
24. Use of a compound, or a pharmaceutically acceptable salt thereof,
according to
any one of claims 1 to 16 in the manufacture of a rnedicament to lower blood
glucose
levels.
25. Use of a compound, or a pharmaceutically acceptable salt thereof,
according to
any one of claims 1 to 16 in the manufacture of a rnedicament for the
treatment of
hyperglycemia.
26. The use of any one of claims 23-25, wherein the medicament is for oral
administration.
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Description

WO 2022/246019
PCT/US2022/029958
-1-
MACROCYCLIC GLUCAGON-LIKE PEPTIDE 1 RECEPTOR AGONISTS
This invention relates to glucagon-like peptide-1 receptor agonists and
therapeutic
uses of the compounds to treat type II diabetes mellitus.
Glucagon-like peptide-1 (GLP-1) is a member of the incretin family of peptide
hormones secreted by intestinal enteroendocrine L-cells. GLP-1 induces the
release of
insulin from beta cells in a glucose dependent manner. However, GLP-1 is
rapidly
metabolized so that only a small percentage of the GLP-1 can be utilized to
induce insulin
secretion. To offset this, GLP-1 receptor (GLP-1R) agonists have been
developed to
enhance insulin secretion as a treatment for type II diabetes mellitus.
The majority of GLP-1R agonists that have been approved to treat type II
diabetes
mellitus are injectable agents. Patients often prefer orally administered
drugs because of
the drawbacks associated with injection such as inconvenience, pain, and the
potential for
injection site irritation.
W02018/109607 discloses certain benzimidazole derivatives, which are described
as GLP-1R agonists. Further GLP-1 agonist compounds are disclosed in
W02019/239371, W02019/239319, W02020/103815, W02020/207474,
W02020/263695, W02021/018023, W02021/081207, W02021/096284,
W02021/096304, W02021/112538, W02021/154796, W02021/160127,
W02021/187886, W02021/197464, CN113480534, W02021/219019, W02021/244645,
W02021/249492, CN113801136, W02021/254470, W02021/259309, W02022/007979,
W02022/031994, W02022/028572, W02022/040600, W02022/042691,
W02022/068772, W02022/078407, W02022/078380 and W02022/078152.
However, there is a need for alternative GLP-1R agonists. In particular, there
is a
need for GLP-1R agonists which can be administered orally. There is especially
a need
for potent GLP-1R agonists which have a favorable toxicology profile and/or a
pharmacokinetic profile which supports once daily dosing.
Accordingly, the present invention provides a compound of the formula:
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-2-
A
ZLZ24
Ni& y2
Z3\\ R5
'Y
)1(1 y6
Formula IX
wherein -A- is -CRaRbCRaRbCRbRb0-, -OCRbRbCRaRbCRaRb-, -OCRbRbCRbRb0-,
-CRaRbCRbRbOCRbRb-, -CRbRbOCRbRbCRaRb-, -CRbRbOCRbRb-,
-CRaRbCRbRb0-, or -OCRbRbCRaRb-;
Ra at each occurrence is independently H, halo, CI-C2alkyl, OH or Ci-GRalkoxy,
Rb at each occurrence is independently H, halo or CI-C2alkyl;
a
a X6 X5
I 7 J,L
=-= 4 X - 8
b X b
11 is or
, wherein a is the point of attachment to linker A,
b is the point of attachment of linker B,
XI, X2, X' and X4 are independently N, CH or CR1, wherein no more than two of
X1, X2,
X' and X4 are N and no more than two of X1, X2, X' and X4 are CR1,
X5 is N, CH or CR1a, X6, X' and X11 are independently N, CH or CR1, wherein no
more
than two of X5, X6, X7 and X' are N and no more than two of X5, X6, X7 and X'
are CRla
or CR1;
R1 at each occurrence is independently CN; halo; Cl-C3alkyl optionally
substituted with
OH, Cl-C3haloalkyl, Cl-C3alkoxy, C3-05cycloalkyl, -S02Ci-C3alkyl;
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-3-
0 Rh
9
Re Re
9
Oy.X/x9 x9 px9
X N
N =
Re"' x9'2:-- /1-X
Re or wherein each X9 is
independently
CH or N and no more than one X9 in the ring is N, each RC is independently
selected
from: H, Ci-C3haloalkyl, halo, C3-05cycloalkyl and Ci-C3alkyl optionally
substituted
with OH, R1' is H, C1-C3haloalkyl, halo, C3-05cycloa1kyl, OH, -NItcRd or Ci-
C3alkyl
optionally substituted with OH;
5- or 6- membered heteroaryl or phenyl wherein the heteroaryl or phenyl is
optionally
substituted with one or two substituents independently selected from: CI-
C3alkoxy, C3-
05cycloalkyl, -CH2-C3-05cycloalkyl, -S02C1-C3alkyl, C4-05heterocyclyl, -CH2-C4-
05heterocyclyl, halo, C1-C3haloalkyl, C1-C3haloalkoxy, CN, -CONRcle, -Nit'Rd
or Ci-
C3alkyl optionally substituted with OH;
Tea is CN; halo; Ci-C3alkyl optionally substituted with OH; Ci-C3haloalkyl; or
Cl-
C3alkoxy;
-B- is -CH20-, -0C112- or -CH2NH-;
y1, Y2 and Y7 are independently N, CH or CR2, wherein no more than one of Y1,
Y2 and
Y7 is N and no more than two of Y1, Y2 and Y7 is CR2;
Y3, Y4, Y5 and Y6 are independently N, CH or CR2, wherein no more than two of
Y3, Y4,
Y5 and Y6 are N and no more than two of Y3, Y4, Y5 and Y6 are CR2;
R2 at each occurrence is independently halo or methyl;
Z1, Z2 and Z3 are independently N, CH or CR3, wherein no more than two of Z1,
Z2 and
Z3 are N and no more than two of Z1, Z2 and Z3 are CR3;
R3 at each occurrence is independently halo; Cl-C4alkyl; -0C4-C6cycloalkyl
optionally
substituted with Cl-C2alkoxy, OH, Ci-C3alkyl or C1-C3haloalkyl; -0C4-
C6heterocycly1
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-4-
optionally substituted with CI-C2alkoxy, OH, CI-C3alky1 or CI-C3haloalkyl; or
CI-
Cztalkoxy optionally substituted with one or two substituents selected from:
CI-C2alkoxy,
OH, -N-RR, -CONWRd, CN, halo or 5- or 6- membered heteroaryl optionally
substituted
with C1-C3alkyl;
,
0
R4 is )70 4N
or =
0
r
N.
N¨S'
N¨N H
R5 is -CO2H, or
RC and Rd are each independently H or C1-C3alkyl;
Rf. is H or Ci-C3alkyl; and
Rg is H, Ci-C3alkyl, Ci-C3haloa1kyl, C3-05cycloalkyl, C(0)Ci-C3alkyl, or C1-
C3alky1C3-
05cycloalkyl;
or a pharmaceutically acceptable salt thereof.
Formula IX includes all individual enantiomers, and mixtures thereof, as well
as
racemates.
In an embodiment, there is provided a compound of the formula:
=
B )(3 fq¨V3 R5
1:-Y2
116
ZI=Z2 I 4
Y Y
-=:y5"
Formula V
or a pharmaceutically acceptable salt thereof.
In an embodiment, there is provided a compound of the formula:
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¨ 5 ¨
II A
3
B.';.; NY 3 I 6 Y1 . Y2 ¨_
¨/ VR5
Z1
\
.I-== =Z2 I 4
Y Y
,
Formula Va
or a pharmaceutically acceptable salt thereof.
In an embodiment, there is provided a compound of the formula:
_\
0 A
y>?''',,',,,i, = '''''..µssr Nr .- ¨ - IN- ''N' sj\ \N
By.) \ (3. A 1 %v2
N i = )_ R
-
li Y ZI=Z2 ,s1(4
,
Formula VI
or a pharmaceutically acceptable salt thereof.
In an embodiment of Formula V, Va and VI, Y4, Y5, Y6 and Y7 are all CH.
1
õ __a
x--
110 õ31 JL
In an embodiment is A':'..X4 b . In one embodiment
is
x2,__x1 a
X b 10 .. ; X1 , X3 and X4 are CH; and X2 is CR'. In an
alternate embodiment,
jc3,.x 4 1
, b
is ; X1 is N; X2 is CR1; and X3 and X4 are CH. In a
further alternate
x2,..xl a
II _ _31
x I
embodiment, is
4----11 ; X1, X3 and X4 are CH; and X2 is N. In a further
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-6-
õ1
xg....õ...Aa
0 alternate embodiment, is )1e 1
; X1 and X4 are CH; X2 is CR1; and X3 is
õi
)(2_,....:Aa
N. In a further alternate embodiment, 0is 13 1
X - 4.-"---
----X b
; X1 and X3 are CH; X2 is
).(2,X1 a
)(3,XjL 4 b
-----
CRi; and X4 is N. In a further alternate embodiment, 0 is
; X1 and X3
are CH; and X2 and X4 are CR1.
In an embodiment, only one of X1, X2, X3 and X4 is N.
a
X6
- X5
In an alternate embodiment, is X---X8 b . In
one embodiment,
a
X6 j)(5
1 7
X - 8
0 is --x b ; X5, X7 and X8 are CH; and X6 is CR1. man
alternate
a
X6 )(5
II embodiment, A is s1,7 JI....,
- 8 b
----X 5
; X- is N; X6 is CR1; and X7 and Xg are CH.
In an embodiment, only one of X', X6, X' and X' is N.
In an embodiment, R1 is CN; halo; Ci-C3a1kyl optionally substituted with OH;
Cl-
C3haloalkyl;
0
Re ReN), Rh
0 0,......S
....--...õ,.,,N,::
R e or wherein each Re is
independently selected from: H, Ci-C3alkyl or halo, and Rh is H or halo;
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-7-
5- or 6- membered heteroaryl or phenyl wherein the heteroaryl or phenyl is
optionally
substituted with one substituent selected from: C1-C3alkoxy, C3 -05
cycloalkyl, -S 02C1-
C3 alkyl, C4-05heterocyclyl, -CH2-C4-05heterocyclyl, halo, -CONR'Rd, -NR'Rd or
Ci-
C3alkyl optionally substituted with OH, wherein RC and Rd are each
independently H or
CI-C3alkyl.
/r4 N
/ ---------
H N ¨N
In an embodiment, R1 is CN, halo, CF3, -CH2OH, , ,
(0..___.,
N----- /7-i
1 0 ,, N N
N 1 ,
N / -----
/ ---- \
---';1"-- -N
H3CO2S 1%1---
1 , \
1 N-----', / /
,
,
0 N 0
/ ---- ')--- HO, HN
N- , N'
F
1 , N --- Nr----1 N , y---..õ
N '., IAN
, , , , ' , ,
0 F
N,_
- - --- - - . ' - . N 0
0 N / I s I -- õ. I = I , -------
-)
--\ -----.'----'4 H2N---------,N,,/
/ or ' .,-- . Preferably, Rl is CN,
,
0
N N
/ ----- H3co2s / ---z--- -/ ---
:--___-õ,
H N N . N
..,
Cl, F, CF3,
, , , , ' ,,--
F
F
) -:- : " . = . , ,- - : - 0
- - - = ..'--- ,
N
...,:_,.._N_,/
=
or ' ."-- . In an embodiment, RI is CN; halo; C1-C3alkyl optionally
Re
0
Re, N ,'"
substituted with OH; C1-C3haloalkyl; ' wherein each Re is
independently
selected from: H or CI-C3alkyl; 5- or 6- membered heteroaryl or phenyl wherein
the
heteroaryl or phenyl is optionally substituted with one substituent selected
from: C1 ¨
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-8-
C3alkoxy, C3-05cycloalkyl, -S02Ci-C3alkyl, -CH2-C4-05heterocycly1, -CONR'Rd or
CI-
C3alkyl optionally substituted with OH, wherein R' and Rd are each
independently H or
N. N
/ ----- / z"------.
HN -N
CI -C3alkyl. In an embodiment, R1 is CN, halo, CF3, -CH2OH,
N1 ------
, H3CO2S N N//-----I ,
N / ----,
/ ------ \
0--'---T;;4- -N N------/,
N I
I \N' / /
' ,/
'C? 0
0 N 0
, HO HN N I ,
N .,N,,_ ,
, ,
,' / ,
or
I, HNiNz's----- -N/ -----:---
NI4
Preferably, R1 is CN, Cl, F, CF3,
H3co2s 0
,
,,,N
or = . In a further embodiment, is R1
is CN, halo, Cl-
C3alkyl or Cl-C3haloa1kyl. In an embodiment, R1 is CN, halo or CF3. In an
embodiment,
R1 is CN or halo.
)(2...,x1 .. a
410 1(3 4
-----Xj-
b
In one embodiment, is ),, ; X1, X3 and X4 are CH; X2 is CR1;
and R1 is CN; halo; Cl-C3alkyl optionally substituted with OH; C1-C3haloalkyl;
0
Re Re 11 Rh
0 ''N" -'= 0,....,,..X
Re or wherein each Re is
independently
,
selected from: H, C,-C3alkyl or halo, and Rh is H or halo;
5- or 6- membered heteroaryl or phenyl wherein the heteroaryl or phenyl is
optionally
substituted with one substituent selected from: Ci-C3alkoxy, C3-05cycloalkyl, -
SO/C1-
C3 alkyl, C4-05heterocyclyl, -CH2-C4-05heterocyclyl, halo, -CONItcRd, -NR'Rd,
or Ci-
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-9-
C3alkyl optionally substituted with OH, wherein R' and Rd are each
independently H or
i----- / --------
.---
HN N
CI-C3 alkyl. Preferably, R1 is CN, halo, CF3, -CH2OH,
NI //------
1 , H3CO2S N N I ,
Ni ------:-
/ ----- \
1
0--'''';'4-
N I ,
/ /
,
,
0
0 N 0
/ y------ HO HN
N ,
-%'' )----- /--:-_-_-. F
1 = N---- N Ns
14-=,,,õ:, .N HN
0 F
N,....
1%1' \N 0
CO __________________ N/
/ or ' =,-- . More
preferably, It' is
71
H3co2s o
/N=------- y
HN , -N = N
7
CN, Cl, F, CF3, , ' , ,'
F
F
)--------.zz- 0
`-,---", *---.----),. ,
N
= or -
x2..... xl a
lb In one embodiment, is
; Xl, X3 and X4 are CH; X2 is CR1;
and R1 is CN; halo; Cl-C3alkyl optionally substituted with OH; C1-C3haloalkyl;
Re
0
'
wherein each RC is independently selected from: H or Cl-C3alkyl; 5- or
6- membered heteroaryl or phenyl wherein the heteroaryl or phenyl is
optionally
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-10-
substituted with one substituent selected from: CI-C3alkoxy, C3-05cycloalkyl, -
SO2Ci-
C3alkyl, -CH9-C4-05heterocyclyl, -CONRcle or CI-C 3 alkyl optionally
substituted with
OH, wherein RC and Rd are each independently H or CI-C3alkyl. Preferably, R1
is CN,
NI':
/ ----s-= _.--- / ----, /1µ1,_ ..___
H N -N N 1 0
1
halo, CF3, -CH2OH, , , =,
H3co2s N N -----=
,0
N
/ ---- \ I = -- ."--y- / "---
-N = N I ,
,
, \ _-- ,= ____,--
,
/ / N
,
,' , ' ,
0
0
H N
, I ,
, N ...,.,4,H
,
,or ,' . More
preferably, R1 is
H3co2s 0
N
N
N a
/ ----
, H N ,
____;24õ
CN, Cl, F, CF3, , , : ,or = .
In a
further embodiment, X1, X3 and X4 are CH; X2 is CR1; and R1 is CN. In a
further
embodiment, X1, X3 and X4 are CH; X2 is CR1; and R1 is Cl. In a further
embodiment, X1,
X3 and X4 are CH; X2 is CR1; and R1 is CF3.
a
0 ,31 A-
In an alternate embodiment, is X4 b ; X' is N; X2 is
CR1; X3 and
X4 are CH; and R1 is CF3.
x2...,X1 a
,31 JL
In an alternate embodiment, 4110 is X4 b ; X1 and X4 are CH; X2 is
CR1; X3 is N; and R1 is CF3.
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1
a
)1(3 jL
0 .X11 b
In an alternate embodiment, is ; X1 and X3 are CH; X2 is
CR1; X4 is N; and R1 is CN.
1
,,____a
0 In an alternate embodiment, is )1(3
4 )( JL
-- b
; X1 and X3 are CH; X2 and
X4 are CR1; and each It1 is independently selected from halo and CN.
Preferably, each R1
is independently selected from F, Cl and CN.
a
X - x5
III In an alternate embodiment, is 1 7 k
; X5, X7 and X8 are CH; X6
is CR1; and ft1 is CN or Cl. Particularly, R1 is CN.
a
x5
41110 In an alternate embodiment, is _17 JI.,
A - 8
--x b 5
; X is N; X6 is CR1; X7 and
X8 are CH; and It' is CN.
In an embodiment, -A- is -CH2CH2CH20-, -OCH2CH20-, -CH2CH2OCH2-,
-CH2OCH2CH2-, -CH2OCH2-, -CH2CH20-, -OCH2CH2- or -CF2CH2OCH2-. In a
particular embodiment, -A- is -CH2CH2CH20-, -CH2OCH2-, -CH2CH2OCH2-,
CH2OCH2CH2- or -CF2CH2OCH2-.
In an embodiment, -A- is -CHRaCHRaCHRb0-, -CHRbOCHRb- or
-OCHRbCHRb0-. In a further embodiment, -A- is -CHRaCHRaCHRb0-. In a particular
embodiment, -A- is -CHRaCHR1CHRb0- and each Ra and Rb is H. In an alternate
embodiment, -A- is -CHRbOCHRb-. In a particular embodiment, -A- is -CHRbOCHRb-
and each Rb is H. In an alternate embodiment, -A- is -OCHRbCHRb0-. In a
particular
embodiment, -A- is -OCHRbCHRb0- and each Rb is H. In a particular embodiment, -
A- is
-CH2CH2CH20-, -OCH2CH20-, -CH2CH2OCH2-, -CH2OCH2-, or -CH2CH20-;
preferably, A- is -CH2CH2CH20- or -CH2OCH2-.
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In an embodiment, -B- is -CELO- or -CMNH-. In an alternate embodiment, B is
-CH90- or -0C112-. In a further embodiment, -B- is -CH90-.
In an embodiment, Y3 is N or CH. In an embodiment, Y3 is N. In an alternate
embodiment, Y3 is CH.
In an embodiment, Y4 is CH.
In an embodiment, Y5 is CH.
In an embodiment, Y6 is CH or CR2.
In an embodiment, Y3 is N; and Y4, Y5 are CH; and Y6 is CH or CR2. In a
further
embodiment, Y3 is N; and Y4, Y5 are CH; and Y6 is CH. In yet a further
embodiment, Y3
is N; and Y4, Y5 are CH; and Y6 is CR2, preferably R2 is F. In an alternate
embodiment,
Y3, Y4, Y5 and Y6 are all CH. In a further alternate embodiment, Y3 and Y6 are
N; and Y4
and Y5 are CH.
In an embodiment, Y1 is CH or CR2.
In an embodiment, Y2 is CH.
In an embodiment, Y7 is CH.
In an embodiment, R2 is F or methyl.
In an embodiment, Y1, Y2 and Y7 are all CH. In an alternate embodiment, Y1 is
CR2; Y2 is CH; Y7 is CH; and R2is F. In a further alternate embodiment, Y1 is
CR2; Y2 is
CH; Y7 is CH; and R2is methyl.
In an embodiment, Y1 and Y2 are both CH. In an alternate embodiment, Y1 is
CR2,
y2 is CH and R2is F. In a further alternate embodiment, Y1 is CR2, Y2 is CH
and R2is
methyl.
In an embodiment, Y4, Y5, Y6 and Y7 are all CH.
In an embodiment, Z1 is CH or CR3.
In an embodiment, Z2 is CH.
In an embodiment, Z3 is CH. In an alternate embodiment, Z3 is N.
In a particular embodiment, Z2 and Z3 are both CH.
In an embodiment, R3 is halo; -0C4-Coheterocycly1 optionally substituted with
Ci-
C3alkyl; or Cl-C4alkoxy optionally substituted with one or two sub stituents
selected from:
Cl-C2alkoxy, OH, -NRfRg, -CONWRd or 5- or 6- membered heteroaryl optionally
substituted with Cl-C 3alkyl; wherein RC and Rd are each independently H or Cl-
C3alkyl,
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is H or CI-C3alkyl, and Rg is H, CI-C3alkyl or CI-C3haloalkyl. In a particular
embodiment, it3 is halo or CI-C4alkoxy optionally substituted with one or two
substituents selected from: Ci-C2alkoxy, OH, or -NRfRg, wherein Rf and Rg are
both CT-T3;
preferably, R3 is F, -OCH3, -OCH2CH2OCH3, OCH2CH2OH or OCH2CH2N(CH3)2.
In an embodiment, le is halo, Ci-Caalkoxy or -C1-C3alkoxyC1-C2alkoxy;
preferably, le is F, -OCH3 or -OCH2CH2OCH3. In an alternate embodiment, R3 is
halo,
CI-C2alkyl or methoxy.
In an embodiment, Z1 is CR3 and It1 is halo; -0C4-C6heterocycly1 optionally
substituted with C1-C3alkyl; or C1-C4alkoxy optionally substituted with one or
two
substituents selected from: C1-C2alkoxy, OH, -NRfRg, -CONWRd or 5- or 6-
membered
heteroaryl optionally substituted with C1-C3alkyl; wherein RC and Rd are each
independently H or Ci-C3alky1, Rf is H or Ci-C3a1kyl, and Rg is H, Ci-C3alky1
or Ci-
C3haloalkyl. In a further embodiment, Z1 is CR3 and R3 is halo or Ci-C4alkoxy
optionally
substituted with one or two substituents selected from: Ci-C2alkoxy, OH, or -
NRfRg,
wherein Rf and Rg are both CH3; preferably, R3 is F, -OCH3, -OCH2CH2OCH3,
OCH2CH2OH or OCH2CH2N(CH3)2. In one embodiment, Z1 is CR3 and R3 is F. In an
alternate embodiment, Z1 is CH. In a further alternate embodiment, Z1 is CR3
and R3 is
methoxy.
N.
' N--
1,1
In an embodiment, R5 is -CO2H. In an alternate embodiment, R5 is H
In an embodiment, there is provided a compound of the formula:
A
rR4
\ 1stB Y3 N R5
Formula VIII
wherein -A- is -CH2CH2CH20-, -OCH2CH20-, -CH2CH2OCH2-, -CH2OCH7CH2-,
-CH2OCH2-, -CH2CH20-, -OCH2CH2- or -CF2CH2OCH2-;
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Xa
X6 X5
8
b 'X b
1111 is or
, wherein a is the point of attachment to linker
A; b is the point of attachment of linker B;
X1, X2, X3 and X4 are independently N, CH or CR1, wherein no more than one of
X1, X2,
X3 and X4 is N and no more than two of X1, X2, X3 and X4 are CR1;
X5 is N or CH, X6, X7 and X8 are independently N, CH or CR1, wherein no more
than one
of X5, X6, X7 and X8 is N and no more than one of X6, X7 and X8 is CR1;
R1 at each occurrence is independently CN; halo; Cl-C3alkyl optionally
substituted with
OH; C1-C3haloalkyl;
0
Re Re N Rh
0 0
Re, N N
R e or wherein each Re is
independently selected from: H, C1-C3alky1 or halo, and Rh is H or halo;
5- or 6- membered heteroaryl or phenyl wherein the heteroaryl or phenyl is
optionally
substituted with one substituent selected from: Cl-C3alkoxy, C3-05cycloalkyl, -
S02Ci-
C3alkyl, C4-05heterocyclyl, -CH2-C4-05heterocyclyl, halo, -CONR`Rd, -NR`Rd or
Ci-
C3alkyl optionally substituted with OH;
-B- is -CH20-, or -CH/NH-;
Y1 is CH or CR2;
Y3 is N or CH;
Y5 is N, CH or CR2;
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R2 at each occurrence is independently halo or methyl;
Z1 is CH or CR3;
Z3 is N or CH;
It3 is halo, -0C4-C6heterocycly1 optionally substituted with C1-C3alkyl, or C1-
C4alkoxy
optionally substituted with one or two substituents selected from: C1-
C2alkoxy, OH,
-NRfRg, -CONWRd or 5- or 6- membered heteroaryl optionally substituted with C1-
C3alkyl;
_cp\N N
'. 0
R4 is , or =
0
NN (?\
N¨S
N¨N
H
R5 is -CO2H, or
RC and Rd are each independently H or Ci-C3alkyl;
Rf. is H or C1-C3alky1; and
Rg is H, Cl-GRalkyl or Ci-C3haloalkyl,
or a pharmaceutically acceptable salt thereof.
In one embodiment of Formula VIII, -A- is -CH2CH2CH20-, -CH2OCH2-,
-CH2CH2OCH2-, CH9OCH2CH9- or -CF9CH2OCH2-.
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,,,õ1
x2.
ID In one embodiment of Formula VIII, is )1(3 1
; X1, X3 and X4 are
CH; and X2 is CR1. In an alternate embodiment of Formula VIII, 4110is
i
)(2,....,..x,õ, a
)1(3_ 4'bx
; X' is N; X2 is CR' ; and X' and X4 are CH. In a further alternate
x2,....xl a
1111 embodiment of Formula VIII ---)( , is 13 jL
X --- 4
b
; X1, X3 and X4 are CH; and X2 is
1
x2,.......x,,....,, a
0 I
---)( b
N. In a further alternate embodiment of Formula VIII, is X31 4-'--- ;
X1 and
X4 are CH; X2 is CR1; and X3 is N. In a further alternate embodiment of
Formula VIII,
1
x2_,,,.......X .._...õ.., a
0
is ; X1 and X3 are CH; X2 is CR1; and X4 is N.
In a further alternate
x2,..x a
411/)(1.3 IL
.-. )e b
embodiment of Formula VIII, is ; X1 and X3 are CH; and X2
and
X4 are CR1.
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-17-
a
X
6 -1x
-
II 1 7
----^ =-=
In one embodiment of Formula VIII,
is X - µ)3 h ; x5, X7 and X8 are
411, CH; and X6 is CR1. In an alternate embodiment of Formula VIII, is
a
X6
- x
X,- X LI 8
-
; X5 is N; X6 is CR1; and X7 and X8 are CH.
N
/ ----'-----
HN
In one embodiment of Formula VIII, R1 is CN, halo, CF,. -CH2OH,
N--%-.-
I 31 ' H C 28
N---..õ
N zN______ 0 / ---
--
¨N
N N 1
1 ,
\N ¨
5
,' ,
, , , ,
0
---;-
N Ni , 1 , ,,.-(:),,--, N
y-..-.,--. HO
N-----'' '
, I
I i N __.-----'
,
0 F
HN --,4'-''-- )------ 7-=----. )-----
1 o
I1 , N \ --- N, N,
I Ni/, ` N __-1V>1 rsiv, Hisl/
0
`-.N''.-= , 0, N
, 0 __ NJ/ ------ 1
0 H 2 N
.." or
, ,
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F
-.'".----- ________________________________ , N/ ---z---
HN/ ----
Preferably, R1 is CN, Cl, F, CF3, , ,
F
F
H3CO2S 0
, .y-'s--,,. 0 ,./"-------------._ `,..."---.
, N N
;
= or ¨ ..
yi
X2.-=" --..---a
11111 In one embodiment of Formula VIII, is
X b
; Xl, X3 and X4 are
N
/ --z------ /N
HN ______________________________________________________________ N
,,,
CH; X2 is CR1; and R1 is CN, halo, CF3, -CH2OH, ,
(0._.Th
N%------', .------
1 , H3CO2S N.-...._
N / ---- N
N I ,
/ ------ 0 ¨N
1 1 ,
, ' \Nr. --,7.' /
''
,,
, '
.0 0
0 N-...._ 0
/ ''''''-'.= HO HN
I ,
, N __Th ,, ..., ,._N.,-.,i), , ,
. '
, ,
,
,
7 7 7 7
-.7 )------- iizz=--------_ F
I, N \--- N, N,
N ---..4 .,,õ,...Nef.: : ,,
I ' ,
0 F
N
----- ,
CO __ N/ N'%
,._,_ I
\ ----- ------;---'4.. H2N1' ==<..õ. _NJ
,r or ' ,,-- .
Preferably, R1 is CN,
,
H3co2s 0
y--"k--,,,
/ sz----.--- / ----- f----..._----,
HN ¨N , N,
Cl, F, CF3, ,, ===
7 1
F
F
)... 0
N,
,'-`-
= or
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X1 is N;
-19-
0
b ; In an alternate embodiment of Formula VIII, is )(
X2 is CRI; X3 and X4 are CH; and R1 is CF3.
x?.........-Xla
411, ,31 I
X b
In an alternate embodiment of Formula VIII, is - X' X3
and X4 are CH, and X2 is N.
1
410
X b
In an alternate embodiment of Formula VIII, is ; XI and X4
are CH; X2 is CRI; X3 is N; and RI is CF3.
0
X b
In an alternate embodiment of Formula VIII, is ; XI and X3
are CH; X2 is CRI; X4 is N; and RI is CN.
x2Xa
X b
In an alternate embodiment of Formula VIII,=is ; X1 and X3
are CH; X2 and X4 are CR' ; and each R1 is independently selected from halo
and CN.
Preferably, each RI is independently selected from F, Cl and CN.
a
X6 x5
40 I 7 X
X - 8
b
5
In an alternate embodiment of Formula VIII, is ; X , X7
and X8 are CH; X6 is CR1; and RI is CN or Cl. Particularly, R1 is CN.
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a
6
X - x5
0 17 11
In an alternate embodiment of Formula VIII, is X ' 8
---X
b ; X5 is N;
X6 is CRI; X7 and Xg are CH; and RI is CN.
In one embodiment of Formula VIII, -B- is -CH20-.
In one embodiment of Formula VIII, Y6 is CH or CR2.
In one embodiment of Formula VIII, R2 is F or methyl.
In one embodiment of Formula VIII, Y3 is N and Y6 is CH or CR2. In a further
embodiment of Formula VIII, Y3 is N and Y6 is CH. In yet a further embodiment,
Y3 is N
and Y6 is CR2, preferably R2 is F. In an alternate embodiment, Y3 and Y6 are
both CH. In
a further alternate embodiment, Y3 and Y6 are both N.
In one embodiment of Formula VIII, Z3 is CH.
In one embodiment of Formula VIII, R3 is halo or C1-C4alkoxy optionally
substituted with one or two substituents selected from: C1-C2alkoxy, OH, or -
NRfitg,
wherein Rf and Rg are both CH3; preferably, R3 is F, -OCH3, -OCH2CWOCH3,
OCII2CII20II or OCII2CII2N(CII3)2.
In one embodiment of Formula VIII, le is or .
Certain compounds of Formula VIII have the following features:
i. -A- is -CH2CH7CH20-, -CH2OCH2-, -CH2CH2OCH2-, CH2OCH2CH2- or
-CF2CH2OCH2-
õ1
1 3 1
---)( b
ii. II is wherein Xl, X3 and X4 are CH, X2 is CRI, and R1 is CN,
H3co2s 0
N-, N y,'__,
/ ----- / .:------.- /-
----.
HN -N N I
,
Cl, F, CF3, /
, , ,
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NN F
0
or
; or wherein X1 is N, X2 is CR1, X3 and X4 are CH, and
R1 is CF,; or wherein X1, X3 and X4 are CH, and X2 is N; or wherein X1 and X4
are CH, X2 is CR1, X3 is N and R1 is CF,; or wherein X1 and X3 are CH, X2 is
CR1,
X4 is N and R1 is CN; or wherein X1 and X3 are CH, X2 and X4 are CR1, and each
R1 is independently selected from F, Cl and CN;
a
X6 --x5
I 7
or is
X8wherein X5, X7 and X8 are CH, X6 is CR1, and R1 is
CN or Cl; or wherein X5 is N, X6 is CR1, X7 and X8 are CH, and R1 is CN;
-B- is -CH20-;
iv. Y1 is CH or CR2, and R2 is F or methyl;
v. Y6 is CH or CR2, and R2 is F;
vi. Z3 is CH;
vii. Z1 is CH or CR3, and R3 is F, -OCH3, -OCH2CH2OCH3, OCH2CH2OH or
OCH2CH2N(CH3)2; and
,F0
viii. K4 is or
In an embodiment, there is provided a compound of the formula:
0 y3 I
1
N = R5
0
Formula VII
or a pharmaceutically acceptable salt thereof.
In a preferred embodiment, there is provided a compound of the formula:
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0 Y3 I
N =
R5
0
Formula Vila
or a pharmaceutically acceptable salt thereof.
In one embodiment of Formula VII and Vila, X2 is CR1 and R1 is CN. In an
alternate embodiment of Formula VII and Vila, X2 is CR1 and R1 is CF3.
In one embodiment of Formula VII and Vila, Y3 is N. In an alternate
embodiment,
Y3 is CH.
In one embodiment of Formula VII and Vila, Y1 is CH. In an alternate
embodiment, Y1 is CR2 and R2 is methyl.
In one embodiment of Formula VII and Vila, R5 is -CO2H. In an alternate
N-N
embodiment, R5 is
In an embodiment, there is provided a compound of the formula:
4
A
73
B )(3 I Y2 NI R5
I Z1Z2
Formula IV
wherein -A- is -CHRaCHRaCHRb 0 -OCHRbCHRaCHRa-, - OCHRb CHRb 0 -,
-CHRaCHRb 0 CHRb - C HRb 0 CHRb CHRa-, -CHRbOCHRb-,
- CHRaCHRb 0 -, or -OCHRbCHRa-;
Ra at each occurrence is independently H, halo, Ci-C2a1kyl or OH;
Rb at each occurrence is independently H, halo or Cl-C2alkyl;
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=
,31 1 _13 11
b b
,s or
, wherein a is the point of attachment to linker
A; b is the point of attachment of linker B;
X1, X2, X3 and X4 are independently N, CH or CR1, wherein no more than two of
X1, X2,
X3 and X4 are N and no more than two of X1, X2, X3 and X4 are CR1;
R1 is CN; halo; Cl-C3alkyl optionally substituted with OH; C1-C3haloalkyl; Cl-
C3alkoxy;
C3-05cycloalkyl; -S02CI-C3alky1;
0
Re Re N 0
Re,N
or Re wherein each Re is independently selected from: H,
CI-C3alkyl optionally substituted with OH, C1-C3haloalkyl, halo and C3-
05cycloalkyl;
5- or 6- membered heteroaryl or phenyl wherein the heteroaryl or phenyl is
optionally
substituted with one or two substituents independently selected from: CI-
C3alkyl
optionally substituted with OH, CI-C3alkoxy, C3-05cycloalkyl, -CH2-C3-
05cycloalkyl,
-S020-C3alkyl, -CH2-C4-05heterocyclyl, halo, C1-C3haloalkyl, Cl-C3haloalkoxy,
CN or
-CONWRd wherein RC and Rd are each independently H or C1-C3alkyl;
-B- is -CH20-, -OCH2- or -CH2NH-;
Y1 and Y2 are independently N, CH or CR2, wherein only one of Y1 and Y2 can be
N;
Y3 is N or CH;
R2 is halo or methyl;
Z1 is N, CH or CR3;
Z2 is CH or CR3;
Z3 is N, CH or CR3;
R3 is halo, Cl-C4alkyl, Cl-C4alkoxy or -C1-C3alkoxyCi-C2alkoxy;
s. 0
R4 i s , or ;and
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N.
N-N
R5 is -CO2H or H
or a pharmaceutically acceptable salt thereof.
In an embodiment, there is provided a compound of Formula IV wherein -A- is
-CHRaCHRaCHRb0-, -OCHRbCHRaCHRa-, -OCHRbCHRb0-, -CHRaCHRbOCHRb-,
-CHRbOCHRbCHRa-, -CHRbOCHRb-, -CHRaCHRb0-, or -OCHRbCHRa-;
Ra at each occurrence is independently H, halo, CI-C2alkyl or OH;
Rb at each occurrence is independently H, halo or Ci-C2alkyl;
X1a x1
,31 I 3
X 4
b
111111 is or
, wherein a is the point of attachment to linker
A; b is the point of attachment of linker B;
Xi, X2, X3 and X4 are independently N, CH or CR1, wherein no more than two of
X1, X2,
X3 and X4 can be N and no more than two of X1, X2, X3 and X4 can be CR1;
R1 is CN, halo, C1-C3alkyl or Ci-C3haloalkyl;
-B- is -CH20-, -OCH2- or -CH2N1-1-;
Y1 and Y2 are independently N, CH or CR2, wherein only one of Y1 and Y2 can be
N;
Y3 is N or CH;
R2 is halo or methyl;
Z1 is N, CH or CR3;
Z2 is CH or CR3;
Z3 is N, CH or CR3;
R3 is halo, Ci-Cialkyl, Ci-C4alkoxy or -Ci-C3alkoxyCi-C2alkoxy;
0 N
0
R4 is or ,and
N¨N
R5 is -CO2H or H
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or a pharmaceutically acceptable salt thereof.
In an alternate embodiment, there is provided a compound of the formula:
3
N \ 02H
1
Formula I
wherein
-A- is -CHRaCHRaCHRb0-, -OCHRbCHRaCHRa-, -OCHRbCHRb0-,
-CHRaCHRbOCHRb-, -CHRbOCHRbCHRa-, -CHRbOCHRb-,
-CHR1CHRb0-, or -OCHRbCHRa-;
Ra at each occurrence is independently H, halo, Ci-C2alkyl or OH;
Rb at each occurrence is independently H, halo or C1-C2alkyl;
Xl, X2, X3 and X4 are independently N, CH or CR1, wherein only one of Xl, X2,
X3 and
X4 can be N and no more than two of Xl, X2, X3 and X4 can be CR1;
R' is CN or halo;
-B- is -CH20- or -OCH2-;
Y1 and Y2 are independently N, CH or CR2, wherein only one of Y1 and Y2 can be
N;
R2 is halo or methyl;
Z1 is N, CH or CR3;
Z2 is CH or CR3;
Z3 is N, CH or CR3; and
R3 is halo, Cl-C2alkyl or methoxy;
or a pharmaceutically acceptable salt thereof.
In an embodiment, there is provided a compound of the formula:
õ ......
Xx4
13 1 0
1Y2 1¨C3 0
2 H
1
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Formula Ia
or a pharmaceutically acceptable salt thereof.
In an embodiment, there is provided a compound of the formula:
1
RI 4,
N
=,,,,..--
Formula II
or a pharmaceutically acceptable salt thereof.
In a preferred embodiment, there is provided a compound of the formula:
1
x2..._x.....,.....,,...,----____0
(/..)
-X4 .-- N
0 N \ i_y-, N ' --CO2H
..-
Formula Ha
or a pharmaceutically acceptable salt thereof.
In an embodiment, there is provided a compound of the formula:
x7--..`s.õ--"-..----------
r-C:07
L.5,Th
,-
N-......!% \>002H
.._
I 1
---
Formula III
or a pharmaceutically acceptable salt thereof.
In a preferred embodiment, there is provided a compound of the formula:
X2---....---- '-'..---'-------- 0 '-''.?
/"... 0
N
\
----' ---. 1
I Y \ 1
Z ¨
-..,...c>
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Formula Ina
or a pharmaceutically acceptable salt thereof.
In an embodiment, -A- is -CHR1CHRaCHRb0-. In a further embodiment, each Ra
and Rb is H.
In an embodiment, X', X' and X' are CH and X2 is CR'. In one embodiment, R1 is
CN. In an alternate embodiment, R1 is Cl.
In an embodiment, -B- is -CH20-.
In one embodiment, Yl and Y2 are both CH. In an alternate embodiment, Yl is
CR2, Y2 is CH and R2 is F.
In an embodiment, Z2 and Z3 are both CH.
In an embodiment, Z1 is CH or CR3. In one embodiment, Z1 is CH. In an
alternate
embodiment, Z1 is CR3 and R3 is F.
In an embodiment, there is provided a compound selected from:
NC NC
N
N 1
N /11, CO2H
CH
,.., F
I __, F F
)
y
CI NC
.---.%----...'','------'-0 0
.---=_,---,1 N T1IIN
I I
N . CO2H
CO2H 0 N
-..
--.
I
.
;
,
NC NC
0 r¨Cc? o i-----
4-Ci
N N
0 IsL N /11, CO2H
.-
0
'
\
;
NC NC
'`--------"-`-------------0 0
I /----Qo r-Q
=,-,...,..,--.1 N N
1 1
0 N N = CO2H HN N N . CO2H
N._ ,-
I I
....-= '. F
F
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NC CN
0
r"--Q
9.)
N 0
I
O N N * CHO2
, -.. N
I I
--- 0 N
0 N III
\ . I
CO2H
;
NC
0 r-Q)
N t
C N
0
N r-4,,:N
0 N N = 1 -
'11
I
N-N
O N., N /II CO2H
I/ Me0 H
,
I
,
NC
0 o-------""--0 0 0
,---Q r---4?
N N
NC
1Z51I 1 /
O N N . CO2H 0 N
Ai, CO2H
1 ,..
1 õ...... F
.
;
,
1-0
I 1., F3C
0
NC /----
Q
0 N
N I
I 0 N N 41 cu2-
H
OJJ N . CO2H I
;-
0
O
\ .
\
,
;
NC 411 0 Br
0
N
I
-- , ../
0 N N CO2H
I CO2H , -...
'... F I
0
\ .
.
,
,
N 0 NC
411 Nr---. ) 0
N
C
f"----
O N N Ali, CO2H
0 N N = CO2H
.
I I
;
.
,
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CN F
r---4-;-;
CO2H
N CO2 H I
0 N
NI *
..--- 0
\ I \
.
,
;
N 0
0 N 7
NC _ j1 I
C
r¨ 0
N N
I i 1
0 N N = CO2H 0 N N .
CO2H
-- ,
I I
,.. --.
¨o . 0
,
?
0
/
;
7 0
H 0-0 F3C N 0 I __
I r-10? .,
=....,,-,i N I ,,
i N
0 N N 41 CO2H
0 N I
N . co
I ; .
I
'..
, 0
\
.
,
N --- , 0 0 ¨\
I r-----Q pi
1 0
002H N
I 0 0 N N .
I
CO2H
\ . ..-'
.
/
/
0 H
., `..., N
I
N ..=
r N \)
0
---4?)
1 1
0 N CO2N 1c) r 1 0 __ \
I,.._ N ,,
;
.
/
NN' \
/ N-N
0 0
1----4O? i - =
= "" Q
N N
0 N 1 0 N 1
CH
CO2H
I N 410
I
;
,
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9, ,0 N
S - I
0 0
N
N 0 N I
i . N
CO2H
I
0 IsL_ N 410, ,
CO2H I N *
---
,., =
;
C?' 0
N ,--
1 N
0 r-O.7 0 N
N , -..
N 410 CH
1 I
0 N ---
C 02 H ;
I
..-- .
1
o
,..
N
0 rQ H
N
N
I
0 Isl,. N 0 CO2H 0 N
JZIX
, I
N . 002H
.
;
=
HO
_-N
0 0
N N
I I
I
CO2H
I N.
CH
---
NC NC 0
0 r--,
0 r----Q
N N
1 N-N I
N AI
0 N. N = / II 0 N
002H
N-N
H .
0
4........-______ N NC
N
N
1 1
0
0 N N = CO2H 0 N
--, N .
I
I F ,.- F
OH
Me0 ¨0
.
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a 0 CN
r"--Q .)==.0
N
I
1
CO2H
N r---1
F 0 NI,, N =N
0 L' I
O N
41, , ==== N= co2H
--
I
---
0-
/0
,
CI CN
0 0 0
N N
t / 1
0 Nõ. N . CO2H 0 N N /11, CO2H
F
0
0
0
0 /
.
/
,
;
F F
NC 0 NC 0
0 Nr-CCI Nr-4;1
i i
0
0 N N = =
CO2H 0 N N
, '..
I I
OH
,--
F 0 0
\__ \
0- .
0
'
/ ;
F
NC 0 0 F
Nr"----Q
N , I N
r--Q
0 1µ1,., NI /0, 0 I L
0 H OH
..--
0 N N N , ---
is
I 0
0
\ . 0
,
0
/
.
,
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NC NC
010 0
Nr-Q 0 0
Nr-Q3
/ 0
AO
I OH I
OH
0 0
F
E N H
,------% F'----' .
;
CI 0 0 CI . 0
1----4(:, r----Q
N N
I / 0
0 .
0 N N =
,
0 H I
---
0 H
0 0
OH 71 -
,
.
,
N
CI 0
0 N
f-----Q I I
I
I / 0
0 11, /
OH N
0 I 0
F OH
NH 0
/
,
/0
;
N ,..
-,.. NC 0 0
r-CC?
N N
I 0 1
, N . CO2H
0
.- F
0 F 0
tINI,.. 0
;
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N ,, NC
.-,
10110 0
NreCi 0
0
, 0 0 N N /11,
0 Nõ, N 41,
0 H I ,..
OH
.,-
o
F .
' 0
N¨
/
;
N"-NJ NC 0
. Nr--
N 1 0
0 N N .fr
OH
0 N
1
--, 0
\ 0
; /
.
,
N
1 I FO 0
141/ o Ns1
*
N
N
0 N 1
N
I iv 0
N ¨
,-, OH
0 F 0
H
N =-=-=N 0 \ /
)-----'2
;
0
/
;
v--- 0\ o
C14 0
'Th
N N..õ-,
N
I OH I / 0
H
0 Nõ N AB,
I 0 I 0
,- F .. F
/¨ 0 /¨ 0
/ /
F N ,
-... o
0 1-----4;)
0 N
N
0 N 1:1 . 0
P F 0 N I
N 0 002N
-,
I N-S=0 I
-. 40 H\< ,- F o
\
o¨ ;
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o.1,...
LI
N
Isl . 0
I ,.. OH; I OH
..---
;
-/-=----i- 0. ...1
N '
01 abr.
0 r-4-0-1
WI 0
reXci
N N
1 0
0
0 N,... N = 0 N(IX N 410,
-.,
I OH 1 ..-
0 H
/ ;
1
O N
0- N "--
,,,N
N 0
0 N N 111, OH
0 N
Nr-K?
I 0 i ' NI = 0
..--- F
OH.
/ ;
0 N -
H 2N \ /
I
0
0 ,¨
rO) 0
N N
0 N N
i \ ,
i ' 1=1 4 0
N S 0
OH.
0 0
`.
.'N
0
0 rO)
0 N
IV 10, 0
0 N .
Ni/111 0 I
----
--- OH.
11 ; and
,
or a pharmaceutically acceptable salt thereof.
In linker A, the left hand terminal group as written is attached to the X ring
and
the right hand terminal group is attached to the Yl, Y2 and Y7 containing
ring. For
example, in the group -CRaRbCRaRbCRbRb0- the oxygen is attached to the Yl, Y2
and Y7
containing ring. In linker B, the left hand terminal group is attached to the
X ring and the
right hand terminal group is attached the Y' containing ring.
The term "halogen" or "halo" refers to fluorine, chlorine, bromine, or iodine.
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The term "Ci-Cnalkyl" refers to a straight, or branched chain saturated
hydrocarbon containing 1 to n carbon atoms. Examples of a Ci-C4alkyl group
include, but
are not limited to, methyl, ethyl, propyl, butyl, and tert-butyl. Examples of
a Ci-C3alkyl
group include, but are not limited to, methyl, ethyl and propyl A C1-C2alkyl
group is
methyl or ethyl.
The term "Ci-Cnhaloalkyl" refers to a Ci-Cnalkyl group, as defined herein,
which
is substituted with one, or more halogen. Examples of C1-C3haloalkyl groups
include, but
are not limited to, trifluoromethyl, difluoromethyl and pentafluoroethyl.
The term "Ci-Cnalkoxy" refers to a straight, or branched chain saturated
hydrocarbon containing 1 to n carbon atoms containing a terminal "0" in the
chain, i.e.,
-0(alkyl). Examples of Ci-C4alkoxy groups include, but are not limited to,
methoxy,
ethoxy, propoxy and butoxy.
The term "Ci-Cnhaloalkoxy" refers to a Ci-Cnalkoxy group, as defined herein,
which is substituted with one, or more halogen. Examples of C1-C3haloalkoxy
groups
include, but are not limited to, trifluoromethoxy, difluoromethoxy and
pentafluoroethoxy.
The term "C3-05cycloalkyl" refers to a monocyclic saturated carbon ring
containing between 3 and 5 carbon atoms. Specifically, it refers to
cyclopropyl,
cyclobutyl or cyclopentyl.
The term "C4-C6cycloalkyl" refers to a monocyclic saturated carbon ring
containing between 4 and 6 carbon atoms. Specifically, it refers to
cyclobutyl,
cyclopentyl or cyclohexyl.
The term -heteroaryl" refers to a monocyclic aromatic ring containing one or
more heteroatoms, preferably selected from: N, S and 0. Examples of 5-membered
heteroaryls include, but are not limited to, pyrazole, triazole and thiazole.
Examples of 6-
membered heteroaryls include, but are not limited to, pyridine and pyridazine.
The term "C4-C6heterocycly1" refers to a 4, 5 or 6 membered monocyclic
saturated ring containing one or more heteroatoms, for example, pyrrolidine.
The term "C4-05heterocycly1" refers to a 4 or 5 membered monocyclic saturated
ring containing one or more heteroatoms, for example, oxetane.
Formula IX encompasses Formulae I, Ia, Ib, II, Ha, IIb, III, Ma, IIIb, IV, V,
Va,
Vb, VI, VII, Vila and VIIb and reference to Formula IX below, for example in
the
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methods of treatment and therapeutic uses, is also to be read as a reference
to each and all
of these sub-formulae.
In another embodiment, there is provided a pharmaceutically acceptable
composition comprising a compound of Formula IX, or a pharmaceutically
acceptable
salt thereof, and at least one of a pharmaceutically acceptable carrier,
diluent or excipient.
In a preferred embodiment, the pharmaceutically acceptable composition is
formulated
for oral administration.
In another embodiment, there is provided a method of treating a patient for
type II
diabetes mellitus, the method comprises administering to the patient in need
of treatment
a pharmaceutically acceptable composition comprising an effective amount of a
compound of Formula IX, or a pharmaceutically acceptable salt thereof, and at
least one
of a pharmaceutically acceptable carrier, diluent or excipient. In one
embodiment, the
pharmaceutically acceptable composition is formulated for oral administration.
Preferably, the patient is a human.
In another embodiment, there is provided a method of treating a patient for
type II
diabetes mellitus, the method comprises administering to the patient in need
of treatment
an effective amount of a compound of Formula IX, or a pharmaceutically
acceptable salt
thereof. In a preferred embodiment, the patient is a human.
In another embodiment, there is provided a method of lowering blood glucose
levels in a patient, the method comprises administering to the patient in need
of treatment
an effective amount of a compound of Formula IX, or a pharmaceutically
acceptable salt
thereof. In a preferred embodiment, the patient is a human.
In another embodiment, there is provided a method of treating hyperglycemia in
a
patient, the method comprises administering to the patient in need of
treatment an
effective amount of a compound of Formula IX, or a pharmaceutically acceptable
salt
thereof. In a preferred embodiment, the patient is a human.
In another embodiment, there is provided a method of treating obesity in a
mammal, the method comprises administering to the patient in need of treatment
an
effective amount of a compound of Formula IX, or a pharmaceutically acceptable
salt
thereof. In a preferred embodiment, the patient is a human.
In another embodiment, there is provided a method of treating nonalcoholic
steatohepatitis (NASH) in a patient, the method comprises administering to the
patient in
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need of treatment an effective amount of a compound of Formula IX, or a
pharmaceutically acceptable salt thereof. In a preferred embodiment, the
patient is a
human
In an embodiment, there is provided a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for use in therapy.
In another embodiment, there is provided a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for use in the treatment of type II
diabetes
mellitus.
In another embodiment, there is provided a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for use in lowering blood glucose
levels.
In another embodiment, there is also provided a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for use in treating hyperglycemia.
In another embodiment, there is provided a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for use in treating obesity.
In another embodiment, there is also provided a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for use in treating NASH.
In an embodiment, there is provided the use of a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for the manufacture of a medicament
for the
treatment of type II diabetes mellitus.
In an embodiment, there is provided the use of a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for the manufacture of a medicament
for
lowering blood glucose levels.
In an embodiment, there is provided the use of a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for the manufacture of a medicament
for the
treatment of hyperglycemia.
In an embodiment, there is provided the use of a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for the manufacture of a medicament
for the
treatment of obesity.
In an embodiment, there is provided the use of a compound of Formula IX, or a
pharmaceutically acceptable salt thereof, for the manufacture of a medicament
for the
treatment of NASH.
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The compounds of Formula IX may be used in simultaneous, separate, or
sequential combination with one or more therapeutic agents. Examples of
additional
therapeutic agents include, but are not limited to, metformin,
thiazolidinediones,
sulfonylureas, dipeptidyl peptidase 4 inhibitors, sodium glucose co-
transporters, and
ketohexokinase inhibitors
In a preferred embodiment, the compound of Formula IX is administered orally.
In a preferred embodiment, the compound of Formula IX is administered once
daily. In
another preferred embodiment, the therapeutic use is in a human.
The term "pharmaceutically acceptable salt" as used herein refers a salt of a
compound of the invention considered to be acceptable for clinical and/or
veterinary use.
Examples of pharmaceutically acceptable salts and common methodologies for
preparing
them can be found in "Handbook of Pharmaceutical Salts: Properties, Selection
and Use"
P. Stahl, et at., 2nd Revised Edition, Wiley-VCH, 2011 and S.M. Berge, et at.,
"Pharmaceutical Salts", Journal of Pharmaceutical Sciences, 1977, 66(1), 1-19.
The term "effective amount" refers to the amount or dose of a compound of
Formula IX, or a pharmaceutically acceptable salt thereof, which, upon single
or multiple
dose administration to the patient, provides the desired effect in the patient
under
diagnosis or treatment. The attending physician, as one skilled in the art,
can readily
determine an effective amount by the use of conventional techniques and by
observing
results obtained under analogous circumstances. Factors considered in the
determination
of an effective amount or dose of a compound include: whether the compound or
its salt
will be administered; the co-administration of other agents, if used; the
size, age, and
general health of the patient; the degree of involvement or the severity of
the disorder; the
response of the individual patient; the mode of administration; the
bioavailability
characteristics of the preparation administered; the dose regimen selected;
and other
relevant circumstances. The compounds of the present invention are effective
at a dosage
per day that falls within the range of about 0.01 to about 15 mg/kg of body
weight.
As used herein, the terms "treating", "to treat", or "treatment", refers to
lowering,
reducing, or reversing the progression or severity of an existing symptom,
disorder, or
condition, such as hyperglycemia, which can include increasing insulin
secretion.
As used herein, the term "patient" includes mammals. The patient is preferably
human.
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The compounds of Formula IX can be formulated as pharmaceutical compositions
administered by any route which makes the compound bioavailable. Preferably,
such
compositions are for oral administration. Preferably the pharmaceutical
compositions are
formulated as a tablet, capsule, or a solution. The tablet, capsule, or
solution can include
a compound of Formula IX in an amount effective for treating a patient in need
of
treatment. Such pharmaceutical compositions and processes for preparing same
are well
known in the art (See, e.g., "Remington: The Science and Practice of
Pharmacy", A.
Adej are Editor, 23 Ed., 2020, Elsevier Science).
The compounds of Formula IX and the pharmaceutically acceptable salts thereof
are useful in the therapeutic uses of the invention, with certain
configurations being
preferred.
Compounds of the present invention include:
je_ 41 0
¨x 3
BN1y2 N \)¨CO2H
Z1=Z2
Formula Ia,
X2<X
x_l 3 4 I
3
B
iy2
\)¨CO2H
1 _2
Z=L
Formula lb.
,3
0 N \
Formula Ha,
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X2 - X1
0
N
I Z3
0 i,y2 N
Z1=- Z2
Formula IIb,
Lm _______________________________________________________
0 N I CO2H
N
Z
Formula Ina,
0 N I N---? )¨CO2H
Y1 Z
Formula Mb,
A
,3
B Y3 I R5
4
Z.1Z2
1 6
Y Y
Formula Va,
A
3
Y2 NI \¨R5
T' IT -Y Z1=Z2
Y4 Y6
y5'
Formula Vb,
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/"... 0
0 y3
R5
Formula VIIa,
X2
0 y3 I NI
Rs
0
Formula VIIb,
or pharmaceutically acceptable salts thereof.
Although the present invention contemplates all individual enantiomers,
mixtures
thereof, and racemates, compounds of Formula Ia, ha, Ma, Va and VIIa and
pharmaceutically acceptable salts thereof, are particularly preferred.
Individual enantiomers may be separated or resolved by one of ordinary skill
in
the art at any convenient point in the synthesis of compounds of the
invention, by
methods such as selective crystallization techniques, chiral chromatography
(See for
example, J. Jacques, el al., "Enantiotners, Racemates, and Resolutions", John
Wiley and
Sons, Inc., 1981, and E.L. Eliel and S.H. Wilen," Stereochemistry of Organic
Compounds", Wiley-Interscience, 1994), or supercritical fluid chromatography
(SFC)
(See for example, T. A. Berger; "Supercritical Fluid Chromatography Primer,"
Agilent
Technologies, July 2015).
A pharmaceutically acceptable salt of the compounds of the invention can be
formed, for example, by reaction of a compound of Formula IX and an
appropriate
pharmaceutically acceptable base in a suitable solvent under standard
conditions well
known in the art (See, for example, Bastin, RI, et al; Org. Process. Res.
Dev., 4, 427-
435, 2000 and Berge, S.M., et al.; I Pharm. Sc., 66, 1-19, 1977).
Certain abbreviations used herein are defined according to Daub G.H., et al.,
"The
Use of Acronyms in Organic Chemistry" Aldrichimica Acta, 1984, 17(1), 6-23.
Certain
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abbreviations are defined as follows: "ACN" refers to acetonitrile; "Boc"
refers to tert-
butyloxycarbonyl; "cAMP" refers to cyclic adenosine-3',5'-monophosphate; "DCM"
refers to di chloromethane or methylene chloride; "DEAD" refers to diethyl
azodicarboxylate, "DIAD" refers to diisopropyl azodicarboxylate; "DIPEA"
refers to
N,N-diisopropylethylamine; "DMF" refers to N,N-dimethylformamide; "DMSO"
refers
to dimethyl sulfoxide; "EC50" refers to the concentration of an agent which
produces 50%
response of the target activity compared to a predefined positive control
compound
(absolute EGO; "EDC" refers to N-(3-dimethylaminopropy1)-N-ethylcarbodiimide
hydrochloride; "ES/MS" refers to electrospray mass spectrometry; "Et0Ac"
refers to
ethyl acetate; "HATU" refers to 1-[bis(dimethylamino)methylene]-1H-1,2,3-
triazolo[4,5-
b]pyridinium 3-oxid hexafluorophosphate; "HEK" refers to human embryonic
kidney;
"HEPES" refers to 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; "h"
refers to
hours or hour, respectively; IrrciF(CF3)ppyi2(dtbpy))PF6 refers to [4,4'--
Bis(1,1-
Clifflethylethyl)-2,2'-bipyridine-N1,N1 lbis[3 ,5 -difitioro-245 -
(trifluoromethi,71)-2-
pyridinyi-Niphenyl-CliriditliTI(III) hexafluorophosphate; "KOAc" refers to
potassium
acetate; "Me0H" refers to methanol or methyl alcohol; "min" refers to minute
or
minutes; "MTBE" refers to methyl tert-butyl ether; "Pd(dppf)C12" refers to [1,
l'-
bis(diphenylphosphino)ferrocene]dichloropalladium(II); PdC12(dtbpf) refers to
[1õ1`--
Bis(di-tert-butylphosphino)ferrocene]dichloropalladi Utn(II); "RT" refers to
room
temperature; "SNAr" refers to nucleophilic aromatic substitution; "TBAF"
refers to
tetrabutylammonium fluoride; "TBS" refers to tert-butyl dimethylsilyl; "TEA"
refers
triethylamine; "TFA" refers to trifluoroacetic acid; "THF" refers to
tetrahydrofuran;
"TMAD" refers to tetramethyl azodicarboxamide; and "TMSCN" refers to
trimethylsilyl
cyanide.
The compounds of the present invention may be prepared by a variety of
procedures, some of which are illustrated in the Preparations and Examples
below. The
specific synthetic steps for each of the routes described may be combined in
different
ways, to prepare compounds of the invention, or salts thereof. The product of
each step
below can be recovered by conventional methods, including extraction,
evaporation,
precipitation, chromatography, filtration, trituration, and crystallization.
The reagents and
starting materials are readily available to one of ordinary skill in the art.
Individual
isomers, enantiomers, and diastereomers may be separated or resolved at any
convenient
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point in the synthesis, by methods such as, selective crystallization
techniques or chiral
chromatography (See for example, I Jacques, et al .,"Enantiomers, 1?acemates,
and
Resolutions", John Wiley and Sons, Inc., 1981, and E.L. Eliel and S.H. Wilen,"
Stereochemistry of Organic Compounds", Wiley-Interscience, 1994). Without
limiting
the scope of the invention, the following preparations, and examples are
provided to
further illustrate the invention.
Scheme 1
yl_ y2
1 yl_ y2
2 Yi¨Y2
Y1=Y2 Br
Br4q-CO2R Step Br--CO2R Br 44 P H Step 3
Y 7 Step Br-74
0 H 0 0
4 /o
Step 4
/eo2R Yl¨Y2 7C co2R
vl=v2 CO2R Y1=Y2 CN
/ 7 / Br-c74 .1E- Br -Y74 0
Step 6 7 5
8 OH OH
6 /0 Step
5/0
or Step 7
0 Yi-Y2 GO2R
>C0'13-c7-
R = methyl, ethyl
OH V, Y2, and Y7 are as defined in Formula IX
9
Scheme 1 shows the synthesis of intermediates 7, 8, and 9 which are used in
the
preparation of compounds of the present invention. In Step 1, the hydroxyl
group of
intermediate 1 is methylated with iodomethane and a carbonate base at elevated
temperature to give methoxy intermediate 2. In Step 2, the ester group of
intermediate 2
is then reduced to alcohol intermediate 3 using NaBH4. The alcohol of
intermediate 3 is
converted to bromide intermediate 4 in Step 3 using PBr3, which is then
reacted with
TMSCN and TBAF in Step 4 to give intermediate 5. The cyano group of
intermediate 5
is treated with sulfuric acid in an alcohol solution at elevated temperature
in Step 5 to
give ester intermediate 6, then the methoxy group is demethylated with BBr3 in
Step 6 to
give intermediate 7. Optionally, intermediate 7 can be converted to a boronate
in Step 7
using KOAc, and Pd(dppf)C12, and either bis(pinacolato)diboron or
bis(neopentyl
glycoloato)diboron at elevated temperature to give boronate 8 or 9,
respectively.
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Scheme 2
F--,-" Z3 Br
- <R4
CO2Me
02N----Z
,o r., /
e
., - ,-
,:3S
iStep 1 4 23 1 Step 5a
,R 1,14
r--- -
FZ3sCNstep 2a F--....-Z3C 2Me., IR4 Step 3a I FINF..-2, 3S
2Mtep 4a H2 CO2Me 1 S
i
O2 NZ 02NZ H2N 02N Z =-_,1
H2
11 Step 2b 12 13 14 15
\R4:-.-"*"-N H2
13
4 H 3 CN NIA
N-11.
R..1.,,Z. Step 3b 4 H H 3 1
R.,,.._,N I NiN Step 4b
R4 N , N)µ1
I i H -a"
H2N -,Zi i
H
021sr.Z12 ,..
02 N z
Step 2c 16 17 18
N-It N-N,
F 3 I )1 Step 3c rI , 1 )4 Step 4c 4 H 3
Nil
1 H ______ .-
N -2- R,..N Z
PG R--NI H2
02 Zi' PG 'Step 5c
02N z 13 02 N-"'Nz
19 20
N-1,
21 i z-ii, N
Z1, Z2, Z3, and R4 are as defined in Formula IX R4 L---
......--
PG = protecting group e.g. SEM (trimethylsilylethoxymethyl) PG
H2N--"-Lµz
22
Scheme 2 shows the synthesis of intermediates 15, 18, 22, and 23 which are
also
used in the preparation of compounds of the present invention. Bromide 10 is
converted
to nitrile 11 in Step 1 using zinc cyanide and a palladium catalyst at
elevated temperature.
Nitrile intermediate 11 is converted to ester intermediate 12 in Step 2a using
thionyl
chloride in an alcohol solution at elevated temperature, then the fluorine is
displaced in a
SNAr reaction in Step 3a with amine 13 and a carbonate base at elevated
temperature to
give intermediate 14. The nitro group is then reduced in Step 4a under an
atmosphere of
hydrogen using Lindlar catalyst (5% Pd) in methanol to give intermediate 15.
Intermediate 15 can be reacted with 2-chloroacetyl chloride using a tertiary
amine base to
give 2-chloromethylimidazole intermediate 23.
To access tetrazole intermediates, intermediate 11 undergoes SNAr reaction
with
amine 13 in Step 2b using an amine base to give intermediate 16, which then is
converted
to tetrazole intermediate 17 using tributyltin azide at elevated temperature
in Step 3b.
The nitro group is then reduced in Step 4b under hydrogen pressure (4 bar)
using a
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palladium on carbon catalyst to give intermediate 18. Alternatively,
intermediate 11 is
reacted with tributyltin azide at elevated temperature in Step 2c to give
tetrazole
intermediate 19, which is then protected in Step 3c on the tetrazole nitrogen
with a group
such as SEM (trimethylsilylethoxymethyl) giving intermediate 20. The fluorine
is
displaced in a SNAr reaction in Step 4c with amine 13 and a tertiary amine
base to give
intermediate 21, then reduction of the nitro group in Step 5c using iron in
acetic acid at
elevated temperature gives protected tetrazole intermediate 22.
Scheme 3
1 ,,i
x2,xx )(2/-"C 02 Et x2---ir,CO2Et
Step la 16 Step 2a
16 HO Y3 W
ic6 e .'
====-= \
-"N X X 'X'4 I -11'.. ...'X4 +
OH OH OH
Y ),=5.
24 25 26 27
i'Step 3a Step lb
x2 xl (:)PG
x2,X1
CO2Et
j6 õ.- 28
)I<3
ii --.
,:=-=-.----
"--x ---X
O 0 Y3-___,r
H,W
29
i Step 2b
14- 16
27
i Step 4a
P, G
T2 xl,,.. .,;,, OH ,X1
3...tep 3b stei.¶ 30.1b
)1(6)(2 H
0 Y3 ....,,µ/V
0 Y3,,..W
30 I; 1 6
Ny5iY 31 32
i: .-y4:y5 jr6
-y5-=
I Step 3c
)(21rXG HO YL;A/ x2,Xx
3(3
+ )7: -µ11,6 Step lc )1(6 Step 2c I
Br 0 Y3,...õW
33 27 34 ): 35 Iir Ls
i -6
G= Br, I; W = Cl, Br
X', X2, X3, X4, Y3, Y4, Y5, and Y6 are as defined in Formula IX;
PG = protecting group e.g. TBS (tert-butyldimethylsily1)
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Scheme 3 shows three routes for synthesizing intermediate 32 which is used in
the
preparation of compounds of the present invention. In Step la in the first
route, halide
intermediate 24 undergoes a Heck coupling with ethyl acryl ate using palladium
acetate
and a carbonate base at elevated temperature to give intermediate 25, which
then
undergoes olefin reduction under hydrogen (40 psi) in Step 2a to give
intermediate 26. In
Step 3a, the alcohol group of intermediate 26 is converted to the bromide
using PBr3, then
reacted with intermediate 27 and Ag2CO3 at elevated temperature to give
intermediate 29.
Reduction of the ester group with LiBH4 in Step 4a gives intermediate 32.
In the second route, intermediate 33 (which can be prepared from intermediate
24
using PBr3) is first reacted in Step lc with intermediate 27 using Ag2CO3 at
elevated
temperature to give intermediate 34, which then undergoes a Negishi coupling
in Step 2c
with bromo-[3-[tert-butyl(dimethyl)silynoxypropyl]zine and a palladium
catalyst at
elevated temperature to give intermediate 35. Deprotection using TBAF in Step
3c then
gives intermediate 32.
In the third route, intermediate 24 undergoes Sonogashira coupling with tert-
butyildimethyl(2-propynyloxy)silane using a palladium catalyst and tertiary
amine base in
Step lb to give intermediate 28, which then undergoes a Mitsunobu reaction
with
intermediate 27 to give intermediate 30. Deprotection using TBAF in Step 3b
and then
hydrogenation of the alkync using platinum oxide under an atmosphere of
hydrogen in
Step 4b gives Intermediate 32.
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Scheme 4
-- --.
x2¨ 113 113
-X1/Stel3 1 I 113 Step 2 x- 47.) Step 3
X 4'
X'X4.--;Th
N N N
X,x4';') 0 0 0 0 0
0
Br
36
37 38 39
1 1 Step 4
-=/-...'0H 1
11
X3 ' Step 5 F Y3 W v2 X
"-- WOH
+ 113
HN ,/, 1(y,5N(6 X,x4-5Th
42 14 116
NH2
W = Cl, Br;
-xl, -x2, -x3, -x4, y3, y-4, y5, and Y6 are as defined in Formula IX
wherein at least one of Y3 and Y4 is N
Scheme 4 shows the synthesis of intermediate 42 which is used in the
preparation
of compounds of the present invention. In Step 1, bromide intermediate 36 is
reacted
with potassium phthalimide at elevated temperature to give intermediate 37. In
Step 2, a
Sonogashira coupling with propargyl alcohol gives alkyne intermediate 38. In
Step 3, the
alkyne of intermediate 38 is reduced with a rhodium catalyst at elevated
temperature
under hydrogen pressure (90 psi) to give intermediate 39. The phthalimide
group is
reacted with hydrazine at elevated temperature to give amine 40 in Step 4,
which then
undergoes a SNAr reaction with intermediate 41 using DIPEA at elevated
temperature in
Step 5 to give intermediate 42.
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Scheme 5
1 2 x1
X2--- === H Step 1 x------. c:C-----orBs Step 2 x"-- ---- ---OTBS
H 0`II3 W
fa 16---X4
0 0 0 H
43 44 45 27
I Step 3
õ1
X.------0 H 2 X1
113 Step 4 v -sir-' ===cirBs
..e ____________________________________________________
je
X, x4'
o,...)r3 w --)(4-
47 r
yty:16 46
T
yty5:16
W = CI, Br;
Xl, X2, X', X4, Y', Y4, Y5, Y6, and Y7 are as defined in Formula IX
Scheme 5 shows the synthesis of intermediate 47 which is used in the
preparation
of compounds of the present invention. In Step 1, intermediate 43 is reacted
with (2-
bromoethoxy)-tert-butyldimethylsilane using a carbonate base at elevated
temperature
The aldehyde of intermediate 44 is reduced in Step 2 using sodium borohydride
to give
alcohol 45, which then undergoes a Mitsunobu reaction with intermediate 27 in
Step 3 to
give intermediate 46. Removal of the tert-butyldimethylsilyl group in Step 4
with TBAF
gives intermediate 47.
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Scheme 6
1 1 1
11
--...x "
CO2Me Step la )(2--xõ,õ¨ c3,x4...i,,c02me OTBS Step 2a
OTBS
0 H 50
48 49
Step 3a HO Y.3_,W
õi I6
-
X-11--0 H le'9'f
3t3 27
---x
0 Y3w
32
H 0 Y3,......,W Step 3b
1 F Y3.,W
i/ 2 X1 3 27 Steop
r 113
lb
41 ic3x2 -.' x----
Step 2b
OPG
0 H -----xl -----x"),
24 0 Y3....õW
0 r...,W
I
34 T4 1,,6
35
....4........y5.,y6
G= Br, I; W = Cl, Br;
Xl, X2, X3, X4, Yl, Y2, Y3, Y4, Y5, and Y6 are as defined in Formula IX
PG = protecting group e.g. TBS (tert-butyldimethylsily1)
Intermediate 32, as described in Scheme 3, can be prepared by the alternative
routes shown in Scheme 6. In Step la, intermediate 48 undergoes a Negishi
coupling
with bromo-[34tert-butyl(dimethypsilyfloxypropyl]zinc and a palladium catalyst
at
elevated temperature to give intermediate 49, which is then reduced to alcohol
intermediate 50 in Step 2a using lithium aluminum hydride. In Step 3a,
intermediate 50
is reacted with intermediate 27 under Mitsunobu conditions to give
intermediate 32.
Alternatively, intermediate 24 is reacted with either intermediate 27 under
Mitsunobu conditions, or with potassium tert-butoxide and aryl fluoride
intermediate 41
to give intermediate 34 in Step lb. Steps 2b and 3b are as described in Scheme
3
(Negishi coupling with bromo-[3-[tert-butyl(dimethyl)silyl]oxypropyl]zinc,
followed by
deprotection to give intermediate 32)
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Scheme 7
x21--X
0 H
)1(6
CE1 EY W
3(6
0' 51
Y1=Y2 CO2R Y Y1=Y2 CO2R
A=
AA ________________________________________________________ Br
Nc.
Y
OH 7 OH
9
Step la Step lb
co2R
CO2R
Y2
0 yBr
I y4-7, 113
jC3
E V3 W
E Y3 W 53
Ys-
52
Step 2a /tep 2b
t
X--"X y71cC 02 R
E Y
54 '11; 6
Step 3
y71T.0O2H
Y31 ly2
1: 16 '
R = methyl, ethyl; D = -CH2-, -0-, or is absent; E = -0-, -NH-;
W = Br, Cl
Xi, X2, X3, X4, Y1, Y2, Y3, Y4, Y5, Y6, and Y7 are as defined in
Formula IX
Scheme 7 shows two routes for synthesizing intermediate 55, which is used in
the
5 preparation of compounds of the present invention. In the first
route, intermediate 51
undergoes a Mitsunobu reaction with intermediate 8 or 9 to give intermediate
52 in Step
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la. In Step 2a, intermediate 52 then undergoes an intramolecular cross-
coupling with a
palladium catalyst to form macrocyclic intermediate 54. In the second route,
intermediate 51 undergoes a Mitsunobu reaction with intermediate 7 to give
intermediate
53 in Step lb, then palladium-catalyzed intramolecular Stille coupling at
elevated
temperature in Step 2b gives intermediate 54. The ester group of intermediate
54 is
hydrolyzed in Step 3 using either aqueous LiOH or a guanidine base in
ACN/water to
give intermediate 55.
Scheme 8
Br OH OTBS ,OTBS
x%)(5 Step 1
)(6--c. 11 Step 2 ,6 Step 3
x. 7 e.J
--, co2Er
se-õ,,,--)---c02_,...Et .ri
ie---xer)--'CO2 Et
56 57 58 59 OH
+
9._.c..õõ.õ.0 H OTBS Step 4 HO Y3 W
X Step 5 yty '53
x8,11,t7x8,..1
27
0,Y3 W 0,,Y3 w
r 61 60
44,6
5.
ylry5)
Step 6
+
x2 j----\-0
r_,_os 2,4,_/1 Y2 co2R 0 Y7 1533)---
8 Step 7 4
V
x
y72g,...1)
0 H itcej 02R
CO2R
0,,ycty
Oy.Y3W I 4 il6
63
62 Y4Z.z.y06
1 Step 8
41-----\.--0
R = methyl, ethyl, W = Cl, Br x x5
x8.._
xs, x6, x7, xs, -,17-1, y-2, y-3, -y-4, y-5, y-6, and Y7 are as defmed y7
`=== 02H
in Formula IX 0,,...V y
0,2
64
Scheme 8 shows the synthesis of intermediate 64 which is used in the
preparation
of compounds of the present invention. In Step 1, intermediate 56 undergoes a
reductive
coupling with 3-bromo-1-propanol at elevated temperature to give intermediate
57, which
then is protected with a TBS group in Step 2 to give intermediate 58.
Reduction of the
ester with lithium borohydride then gives alcohol intermediate 59 in Step 3,
which then
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undergoes a Mitsunobu reaction with intermediate 27 in Step 4 to give
intermediate 60.
Removal of the TBS protecting group with TBAF gives intermediate 61 in Step 5,
which
then undergoes a Mitsutnobu reaction with intermediate 8 in Step 6 to give
intermediate
62. In Step 7, intermediate 62 is cyclized using a palladium catalyst and
potassium
phosphate to give intermediate 63, which is then hydrolyzed in Step 8 using a
using either
aqueous LiOH or a guanidine base in ACN/water to give acid intermediate 64.
Scheme 9
I I
I
Ho2cy-L Step 1a HO Y2 y2 vi
i7 11+ I ,2,x1
'- X2------.Br Step 2
y y
[-, ii -.... 11
7 .
y,ry
1 x----x4---co2m.
----x , ¨02Me ---r--'
Br Br
Br
65 66 67
68
i 1
2-Xi CO2M e x2_,......xco2Me x2õ:__X
OH
_31 I 13 I
)1(3-X4-1Th Step lb A =-= 4
I. --x -Th Step 2b - .4c
x----x
0,.,(3w
Br HO Y3 W 71 Li kli,6
69 Y, 4 70 44 µ1,,6
-..-y5-
Step 3a
...-y5- 27
I
Step 3 Br----'-(LY2
17 I 1
72 Yy-- Y 4,
co2Et I Br
I
1 1 Y2 ,1
.,...__0/-4-: ¨ 1
Step 4a x2,---"-,------02
7 i.1 , Step 5 )1(3 4 I V Y I I
=--- ..,_ ,,e3s, 4 7 1 1
=)( 4 ---X Br
's----X T
0,Y5 Br 0,3 W HO Y3 W OH
Br
,1(6
la
74 y,,..y,:, 4õ,y54 .16
75 Y
w 73
27
Step 6
xi
i
Tyr
)1(3 =)C)
----::-X
Ste 7
0 y3õ..1):: 1,1y2 CO2Et p "-:---x
0 Y3y4yi-Y
-.. y
76 ' &" 77 -=)(5-
W = Cl, Br
xi, x2, x3, x4, yl, y2, y3, y4, Y5, Y6, and Y7 are as defined in Formula IX
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Scheme 9 shows the synthesis of intermediate 77 which is used in the
preparation
of compounds of the present invention, and two synthetic routes to the common
intermediate 74 are shown.
In the first route to intermediate 74, in Step la, acid intermediate 65 is
reduced
using borane-dimethylsulfide complex to give alcohol intermediate 66, which
then
undergoes a reaction with sodium hydride and bromide intermediate 67 in Step
2a to give
intermediate 68. In Step 3a, ester reduction with lithium borohydride gives
intermediate
73, which then undergoes a Mitsunobu reaction with intermediate 27 to give
intermediate
74 in Step 4a.
In the second route to intermediate 74, in Step lb, alkyl bromide intermediate
69
is reacted with intermediate 27 using silver carbonate at elevated temperature
to give
intermediate 70. In Step 2b, the ester of intermediate 70 is reduced with
lithium
borohydride to give alcohol 71, which is then reacted with alkyl bromide 72
using
potassium tert-butoxide to give intermediate 74 in Step 3b.
In Step 5, intermediate 74 is coupled with ethyl diazoacetate using a
palladium
catalyst at elevated temperature to give intermediate 75. In Step 6, an
intramolecular
Stille coupling with a palladium catalyst at elevated temperature gives
intermediate 76.
Alternatively, Step 6 is accomplished by one-pot coupling with bis(neopentyl
glycolato)diboron using a palladium catalyst and potassium pivalatc and
intramolccular
cross-coupling to give cyclic intermediate 76. Intermediate 76 is then
hydrolyzed using
either aqueous LiOH or a guanidine base in ACN/water in Step 7 to give acid
intermediate 77.
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Scheme 10
Step 1 a x2X Step 2a
113 -3" 113
X..,x4-*.-"C 02R
0 H 80
Y3
78 79 F W,)(
V6
41 Step 3a
14'%-y5=--
1
1
0 Y3 X
Step o Y3 w
34
81
Step 4
o H x2,-X 0
=it
-"X
-"X
Step 5
83 82
NI/4.,y5j6
G = Cl, Br, I; W = Cl, Br
R = methyl, ethyl;
XI, X2, X3, X4, Y3, Y4, Y5, and Y6 are as defined in Formula IX
Scheme 10 shows the preparation of intermediate 83, which is used in the
preparation of compounds of the present invention. Two routes to common
intermediate
81 are shown. In the first route, intermediate 78 is coupled with 2-[(E)-2-
ethoxyviny1]-
4,4,5,5-tetramethy1-1,3,2-dioxaborolane using a palladium catalyst and
carbonate base at
elevated temperature in Step la to give intermediate 79. In Step 2a the ester
is treated
with a reducing agent such as diisobutylaluminum hydride to give alcohol 80,
then
reacted with aryl fluoride 41 using a strong organic base such as potassium
tert-butoxide
to give intermediate 81. In the second route, intermediate 34 (see Scheme 3)
is first
coupled with (E)-1-ethoxyethene-2-boronic acid pinacol ester using a palladium
catalyst
and an inorganic base at elevated temperature to give intermediate 81 in Step
lb.
In Step 4, intermediate 81 is treated with HC1 in an organic solvent to give
aldehyde 82, which is then reduced with NaBH4 in Step 3 to give alcohol
intermediate 83
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Alternatively, intermediate 81 can be converted to intermediate 83 in one step
using
mercuric acetate and NaBH4.
Scheme 11
yl y2 Y1=Y2 Step 1 Y1=Y2 c 2R
Step 2 CO2R
x x
84 85 86 Br
X = Br, I; R = methyl, ethyl;
Y1, Y2, and Y7 are as defined in Formula IX
Scheme 11 shows the preparation of intermediate 86, which is used in the
preparation of compounds of the present invention. In Step 1, aryl iodide 84
undergoes
Negishi coupling with (2-ethoxy-2-oxoethyl)zinc(II) bromide and a palladium
catalyst at
elevated temperature. In Step 2, intermediate 85 undergoes photochemical
bromination
with N-bromosuccinimide in a flow reactor to give bromide intermediate 86.
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Scheme 12
I Yji
y X
---)( 3 ,,,,
yi y2 ck-rr ' , ..--
" Step lc
X¨y7.¨i ),,y511/6
Step lb 88
87 Br
1
1'6--- O2R )(4 Step la CY2
___________________________________________________ 30. i I...õ,:-.,yi
I i
o Y3 w yl.y2 CO2 R
Y A
--)(
86 0 Y3,,,w
:rrzyr 89
I Step 2a
Step 2b
X47' Y7 C 2 R
0,1Y3 .vy2
5,
91 ''')LNI(6 Y '.
'Ilr''''''"."""=s...õ.4....... 1 I
Step 2c
I
)1(6 iD..... Step 3 ----x
x2--
16
X)(4%-"--1 y7CO2H
0Y3...seAyiy2
92 44 1,6
X = Br, I; W = Cl, Br
R = methyl, ethyl;
XI, X2, X3, X4, Yl-, Y2, Y3, Y4, Y5, Y6, and Y7 are as defined in Formula IX
Scheme 12 shows multiple routes to the preparation of intermediate 92, which
is
used in the preparation of compounds of the present invention In Step la,
intermediate
5 83 is reacted with intermediate 86 using 2,6-di-tert-butylpyridine
and silver
triflluoromethanesulfonate to give intermediate 89. Alternatively,
intermediate 83 can be
first reacted in Step lb with alkyl bromide 87 under similar conditions to
Step la to give
intermediate 88, which then undergoes Negishi coupling in Step lc with (2-
ethoxy-2-
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oxoethyl))zinc bromide and a palladium catalyst at elevated temperature to
give
intermediate 89. In Step 2a, intermediate 89 undergoes a palladium-catalyzed
intramolecular Stille coupling at elevated temperature to give intermediate 91
Alternatively, in Step 2b, the bromide 89 is converted to the boronic ester
intermediate 90
by cross-coupling with bis(pinacolato)diboron, Pd(dppf)C12 and potassium
acetate at
elevated temperature, which then in Step 2c is cyclized via intramolecular
cross-coupling
with a palladium catalyst to form macrocyclic intermediate 91 (Steps 2b and 2c
can be
performed as a single reaction step). Finally in Step 3 the ester is
hydrolyzed using either
aqueous LiOH or a guanidine base in ACN/water to give acid intermediate 92.
Scheme 13
Br .-0 H
Br X6--"Lx5 X 5
X6---Lx5 Step 1 -
Step 2 117
+ Ho,.e.,,Y3 W
_)...
X
Yrty9.-1
OH 95
i
93 27
1 Y2 CO2 R
Step 3
Br4µle.' _, / -
7 /
96
Br
Br
xtc()(YB-o
1 Step 4 x9----cc)
Ni
...4_
2X
602R
117 x cj....1
602RY
f ya
yft...y5:
98
97z.,....y5.2
Step 5 /
x6 Co
1 x5 q \ x5
X
Xx a. Step 6 N.,
A1
CO2H
y7 "*===== C 02R -,..-
03,w)Lyi,
99 14
Y,z.y5..-V6 100
R ¨ methyl, ethyl; W ¨ Cl, Br;
X5, X6, X7, X8, Y1, Y2, Y3, Y4, Y5, Y6, and Y7 are as defined in Formula IX
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Scheme 13 shows the preparation of intermediate 100, which is used in the
preparation of compounds of the present invention. Intermediate 93 is first
coupled to
intermediate 27 using Mitsunobu conditions to give intermediate 94 in Step 1
In Step 2,
intermediate 94 is coupled with bromoethanol using nickel and iridium
catalysts and
irradiating the reaction under blue light (456 nm) to give intermediate 95.
Alternatively,
intermediate 95 is prepared in a manner analogous to the synthetic routes
shown in the
preparation of intermediate 83 in Scheme 10, starting with intermediate 93 in
place of
intermediate 78. In Step 3, intermediate 95 is reacted with intermediate 96
using 2,6-di-
tert-butylpyridine and silver triflluoromethanesulfonate to give intermediate
97. In Step
4, bromide intermediate 97 is converted to the boronic ester intermediate 98
by cross-
coupling with bis(pinacolato)diboron, Pd(dppf)C12and potassium acetate at
elevated
temperature. In Step 5, intermediate 98 then undergoes an intramolecular cross-
coupling
with a palladium catalyst to form macrocyclic intermediate 99. Alternatively,
intermediate 97 is converted in one step to intermediate 99 via intramolecular
Stille
coupling using hexamethylditin and a palladium catalyst at elevated
temperature.
Intermediate 99 is then hydrolyzed using either aqueous LiOH or a guanidine
base in
ACN/water in Step 6 to give acid intermediate 100.
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Scheme 14
Eto2c Eto2c Eto2c
----," ¨711 1-Y7)-LBr 7 Br Br
Step 1 Y2---y1 Step 2 Y2`-y1 Step 3
-' 7
I
I J( 7j(Br
101 102 Et0
103 HO
104
õi 1 1
X.. 16
x2..---X)H Step 4 x2,X,, 0 H Step 6
113 - Step 5 113
-111"" ".X47 X--.'-X4
X---"X4' 1
0 OH H 0,Y3 W 0.,.."3 W
43
I
y 105
106
27
1
x2-Xx; CO2Et
1 Y2
'-X Y7 CO2Et.4 Step 7
----f
108
14:y2:1 ji,6 . X4-5Th Br
0.......Y3 W
1 1 107 Step 8
1=-Y---
x2-XICI)
ip
X Y7 CO2H
0 ryNyi,
....-V.4:ylsr_y
109
W = Cl, Br; PG = protecting group e.g. SEM (trimethylsilylethoxymethyl)
xl, x2, x3, x4, yl, y2, y3, y4, y5, y6, and Y7 are as defined in Formula IX
Scheme 14 shows the preparation of intermediate 109, which is used to prepare
compounds of the present invention. In Step 1, intermediate 101 undergoes
radical
bromination with N-bromosuccinimide at elevated temperature, then reacted with
trimethylsilyl cyanide and TBAF, then treated with sulfuric acid in aqueous
Et0H at
elevated temperature to give ester intermediate 102. In Step 2, intermediate
102 is
coupled with (E)-1-ethoxyethene-2-boronic acid pinacol ester using a palladium
catalyst
and a carbonate base to give intermediate 103, which is then converted to
alcohol 104 in
Step 3 using mercuric acetate and sodium borohydride. Separately, aldehyde
intermediate
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43 is first protected e.g. with a SEM group and then reduced with sodium
borohydride in
Step 4 to give intermediate 105. In Step 5, intermediate 105 undergoes a
Mitsunobu
reaction with intermediate 27 to give intermediate 106. Intermediates 104 and
106 are
coupled via Mitsunobu reaction to give intermediate 107, which then undergoes
intramolecular cyclization in Step 7 in a manner analogous to Step 2a (one-pot
Stille
coupling) or Steps 2b and 2c (borylation followed by Pd-catalyzed cross-
coupling) in
Scheme 12 to give intermediate 108. Ester hydrolysis using either aqueous LiOH
or a
guanidine base in ACN/water in Step 8 then gives intermediate 109.
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Scheme 15
HO Y.3 ..,,W
0
Step la le
----x
Br -=-=-=-al. 3 Step 2a
'`...,...,,,44.,
1441 110 '-4-. 3_6
) (
*2-- G X----)()--'0 H
1
____Step lb x2-- X3
0 Y3,,,W 112
34 ----x
o Y3...,w
'T-I -----
,y5_,Y6
111 1;1:- 1.6
Y 1 Step 3
Et020 1
x2,x
Br
HO I: 1.
104
Step 4
113
002Et
,xl
xi
x2--
step
0 Y3,yi, 0 H .41 Step
--x) L _.3 y6
115
ovY1.--RW 114
..y.. -,-..Y"
G= Br, I; W = Cl, Br;
Xl, X2, X3, X4, Yl, Y2, Y3, Y4, Y5, Y6, and Y7 are as defined in Formula IX
Scheme 15 shows the preparation of intermediate 115, which is used to prepare
compounds of the present invention. In Step la, alkyl bromide intermediate 144
is reacted
with intermediate 27 using silver carbonate at elevated temperature, followed
by ester
reduction using Red-Al in Step 2a to give intermediate 112. Alternatively,
intermediate
34 is carbonylated in Step lb using potassium formate and a palladium catalyst
to give
intermediate 111, followed by reduction of the aldehyde in Step 2b using NaBH4
to give
intermediate 112. In Step 3, the alcohol of intermediate 112 is converted to
the alkyl
bromide using CBr4 and triphenylphosphine to give intermediate 113. In Step 4,
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intermediates 113 and 104 are reacted with silver trifluoromethanesulfonate to
give
intermediate 114. Intermediate 114 then undergoes intramolecular cyclization
in Step 5
in a manner analogous to Step 2a (one-pot Stille coupling) in Scheme 12, or in
Steps 5
and 6 in a manner similar to Steps 2b and 2c (borylation followed by Pd-
catalyzed cross-
coupling) in Scheme 12. The resulting ester is hydrolyzed in Step 7 using
either aqueous
LiOH or a guanidine base in ACN/water to give intermediate 115.
Scheme 16
1 FE F F
Step 1 .2 y, 1
Step 2 x2,-
CO2M e
CO2Me
X3 X3 -3w 11
X
--X --- 118
Br
116 117
Step 3 1H0 Y.3W
I Y2 1 1
F F 27
F F
...'-' ff' Step 5 X2lX1 OH Ste 4 X2 X
I P 7., ___________
X3
r----y Br .it_1
CO2Me
-`X
Br 0 Y.3..,W
124 0
Y3,,,IN
120 Ti 1 6
-y5.;----Y
119 ii:y0(6
\--- I
Y2 r
I Eto2c i
'`---' . Br I 7
F F 0 Step 6 1 F F ry
Br ----
122
121
Step
.iie---
F F
8
)ez..... 4
y7
0
X -Thy
0 Ne3 r Y2 OH
123 Tt.sy.5._.3y6
W = Cl, Br;
X', X2, V, X', Y', Y2, Y3, Y", Y5, Y6, and Y7 are as defined in Formula IX
Scheme 16 shows the preparation of intermediate 123, which is used in the
preparation of compounds of the present invention. In Step 1, intermediate 116
is reacted
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with methyl bromodifluoroacetate and copper to give intermediate 117, which is
then
brominated photochemically in Step 2 using N-bromosuccinimide in a flow
reactor to
give alkyl bromide intermediate 118. In Step 3, intermediate 118 is reacted
with
intermediate 27 using a phosphate base at elevated temperature to give
intermediate 119,
which then undergoes LiBH4 reduction in Step 4 to give alcohol intermediate
120. In
Step 5, intermediate 120 is treated with NaH and reacted with intermediate 124
to give
intermediate 121. In Step 6, intermediate 121 undergoes Negishi coupling with
(2-
ethoxy-2-oxo-ethyl)zinc bromide and a palladium catalyst at elevated
temperature to give
intermediate 122. Intermediate 122 then undergoes intramolecular cyclization
in Step 7
in a manner analogous to Step 2a (one-pot Stille coupling) or Steps 2b and 2c
(borylation
followed by Pd-catalyzed cross-coupling) in Scheme 12. Ester hydrolysis using
either
aqueous LiOH or a guanidine base in ACN/water in Step 8 then gives
intermediate 123.
Scheme 17
x2,-X 0 H yi y2 1
+ Br
/ CI )1(6 Step 1
y4"Br
0 125 0 Y3_,AA/
83 Br
1-*
y16
126
I
Step 2
j
128
Step 3
B (6 CI
X y7K Y7 =-
-)C
1
127 Ti y6
W= Cl, Br;
XI, X2, X3, X4, Y1, Y2, Y3, Y4, Y5, Y6, and Y7 are as defined in Formula IX
Scheme 17 shows the preparation of boronate intermediate 128, which is used in
the preparation of compounds of the present invention. In Step 1, intermediate
83 is
reacted with alkyl bromide intermediate 125 using silver
trifluoromethanesulfonate to
give intermediate 126, which then undergoes intramolecular cyclization in Step
2 in a
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manner analogous to Step 2a (one-pot Stille coupling) or Steps 2b and 2c
(borylation
followed by Pd-catalyzed cross-coupling) in Scheme 12. Finally, in Step 3
intermediate
127 is coupled with bis(pinacolato)diboron using a palladium catalyst and
potassium
acetate at elevated temperature to give boronate intermediate 128.
Scheme 18
CO
)
rR4
y ,-, .-µ B....Y3 __ Y2 N / \ t'icii ,(1=-
L z` N-S1:13
B.,,,3Vc,yr R4 I 4
I 4 3 Ncy5,
IX" H
\ __ <
N_Th,,ZCO2Me
129 ciN---C-z1-12 t Step 4a
Step la 23
11110 A 1¨R4
OF r-R4 Step 3a 13
Y7 ..2 I___%-/ 2
,,ir .kyty N = 1 co2Ei
_cio2Kie Y4 5. IX'
Y 1
143 \tep 2a
4
41111 A r-R
H N 3
Z 411 A H mr-R4
Y.\7\'''rCO2H H2N¨,¨ Y
1¨0O2Me
13,...,....y3Vc y2 0 , 1z2L-0O2Me
13,sY3Vcyty _
1:
S I 4
Step le 132
04
130 r s
Hz3
Step lb H2N /1 __
PG
18 or 22
0 A H H N7-414
y7 ...N>..-.....rrN? ---z3 ___*. 0 A R
Ni>.....r.....r.Nr- 43
1 IV -N--
Bõ,..Y3,A ty2 0 Step 2b B
sz2"--(' jl c -,t.
ty
yl 4 Y N-N otep 3 u I 4 Y
H
131 PG
X. A, B, Y', Y2, Y3, Y4, V, Y6, Y7, R4, Z', Z2, and Z3
are as defined in Formula IX
PG = H or protecting group e.g. SEM (trimethvlsilyloxymethyl)
Scheme 18 shows the preparation of compounds of the present invention via a
number of different routes from either intermediate 129 (a general formula
which
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encompasses intermediate 128) or intermediate 130 (a general formula which
encompasses intermediates 55, 64, 77, 92, 100, 109, 115 and 123).
To prepare acid compounds of Formula IX', intermediate 129 is coupled with
chloromethylimidazole intermediate 23 in Step la using a palladium catalyst
and a
phosphate base at elevated temperature to give intermediate 143, then in Step
3a the ester
is hydrolyzed using either aqueous LiOH or a guanidine base in ACN/water at
elevated
temperature to give acid of Formula IX'. Alternatively, acid intermediate 130
is coupled
with intermediate 15 in Step lc with an amide coupling reagent such as EDC or
HATU to
give intermediate 132. Intermediate 132 is then cyclized in Step 2a using
acetic acid at
elevated temperature to give intermediate 143, then the ester is hydrolyzed as
described in
Step 3a.
Compounds of the Formula IX" are prepared in Step 4a by coupling an acid of
the
Formula IX' with cyclopropylmethanesulfonamide using EDC and 4-
dimethylaminopyridine.
Compounds of the Formula IX¨ are prepared by coupling intermediate 130 with
intermediate 18 (without a tetrazole nitrogen protecting group such as SEM) or
22 (with a
tetrazole nitrogen protecting group) using HATU in Step lb to give
intermediate 131,
then in Step 2b cyclization with acetic acid at elevated temperature (then if
needed, Step
3b ¨ tctrazole deprotection e.g. using TBAF to remove a SEM group) gives the
tctrazolc
compound of Formula IX¨.
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Scheme 19
R4
,
,.... I 3
HN2.......0O2Me
Isi2,N
02
F Step 1 c
137 HO-R
136
3 R4
F OZ C 2Me Step la F 3
.õ, HNI Z CO2Me
--- -"--T-::1 CO2Me H2N---"R4 ,,õ 3
13
02NIN.Nr-2- __1.,, I _Ii.
N
X --ry
02N
02N
1330H Ho_R Step 2 138
o'sR 135 0
136.....õ...,/.. 'IR
Step lb
Step 3
F Z3 CO2Me I
-- JR4
1
r _3
02NX))
F
134 0 HN CO2Me
Y7 '',- CO2H H2N;.y.:-1.1
139
BY3.cyr,Y2 0-R
I 4
13,40
II A rR4 Steps 4, 5,and 6
Y7 i_____
Z3
13,..,.,3)c y2 N /
¨CO2H
1 Y
1 4 = 6 l'' ¨ Z
q
pc!!
R
X, A, B, Y1, Y2, Y3, Y4, Y5, Y6, Y7, R4, Z2, and Z3 are as defined in Formula
IX
-0-R is a subset of R3 as defined in Formula IX
Scheme 19 shows the preparation of compounds of the present invention of
Formula IX". Intermediate 133 undergoes a Mitsunobu reaction in Step la with
alcohol
intermediate 136 to give 135, which then undergoes a SNAr reaction in Step 2
with amine
13 and a tertiary amine base at elevated temperature to give intermediate 138.
Alternatively, difluoro aryl intermediate 134 first undergoes SNAr reaction in
Step lb
with alcohol 136, which is first treated with NaH and then reacted with
intermediate 134
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at elevated temperature to give intermediate 135. A second alternative gives
intermediate
138 in Step lc by SNAr reaction with intermediate 137 and 136 in a manner
similar to
Step lb. The nitro group of intermediate 138 is reduced e.g. with hydrogen gas
and
palladium on carbon to give aniline intermediate 139 in Step 3. Then, in a
manner
analogous to Steps lc, 2a, and 3a in Scheme 18, compounds of Formula IX" are
prepared in three steps from intermediates 139 and 130. If the "-O-R" group
depicted in
Scheme 19 bears a protecting group, e.g. a Boc group on a nitrogen or tea-
butyldimethylsilyl group on an oxygen, that protecting group can be removed as
a last
step (e.g. using TFA to remove a Boc group or TBAF to remove a tert-
butyldimethylsilyl
group).
Scheme 20
RO
1
RO
rR4
I s'X4 y7
3
14 116 Y
14 116 Y
140 \ 142
D-B(OR)2
/D-W
D,r¨X1
X 4
z3
14 =Z
141
W= Cl, Br, or I
X1, X2, X3, X4, Y1, Y2, Y3, Y4, Y5, Y6, Y7, Z1, Z2, Z3, R4, R5, A, and B are
as defined in
Formula IX
D = 5- or 6-membered optionally substituted aryl or heteroaryl group
R = H or alkyl groups joined to form a cyclic boronic ester
Scheme 20 shows the preparation of compounds of the present invention from
halide intermediate 140. Intermediate 140 undergoes a cross-coupling reaction
(e.g.
Suzuki) with an optionally substituted 5- or 6-membered aryl or heteroaryl
boronic acid
or boronic ester using a palladium catalyst and an inorganic base at elevated
temperature
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to give 141. Alternatively, 140 can be converted to boronate 142, for example
using
tetrahydroxydiboron and a palladium catalyst at elevated temperature giving
142 as a
boronic acid, which then undergoes a cross-couping reaction (e.g. Suzuki) with
an
optionally substituted 5- or 6-membered aryl or heteroaryl halide to give 141.
These
steps can be carried out on either protected or unprotected versions of R5,
for example an
ester can serve as a protected functional group which can be hydrolyzed to
give R5 = -
CO2H.
Preparations and Examples
LC-ES/MS is performed on an AGILENT' HP1200 liquid chromatography
system. Electrospray mass spectrometry measurements (acquired in positive
and/or
negative mode) are performed on a Mass Selective Detector quadrupole mass
spectrometer interfaced to an HPLC which may or may not have an ELSD. LC-ES/MS
conditions (low pH): column: PHENOMENEX GEMINI NX C18 2.0 x 50 mm 3.0
!Am, 110 A; gradient: 5-95% B in 1.5 min, then 95% B for 0.5 min column
temperature:
50 C +/-10 "V; flow rate: 1.2 mL/min; 1 IAL injection volume; Solvent A:
deionized
water with 0.1% HCOOH; Solvent B: ACN with 0.1% formic acid; wavelength 200-
400
nm and 212-216 nm. If the HPLC is equipped with an ELSD the settings are 45 C
evaporator temperature, 40 C nebulizer temperature, and 1.6 SLM gas flow
rate.
Alternate LC-MS conditions (high pH): column: Waters xBridge C18 column 2.1
x50
mm, 3.5 ilnl; gradient: 5-95% B in 1.5 min, then 95% B for 0.50 min; column
temperature: 50 C +/-10 C; flow rate: 1.2 mL/min; 14 injection volume;
Solvent
A: 10 mM NH4HCO3 pH 9; Solvent B: ACN ; wavelength: 200-400 nm and 212-216nm;
if had ELSD: 45 C evaporator temp, 40 C nebulizer temp, and 1.60 SLM gas flow
rate.
Preparation 1
Methyl 4-bromo-5-fluoro-2-methoxy-benzoate
0
CO2Me
Br
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To a mixture of methyl 4-bromo-5-fluoro-2-hydroxy-benzoate (10.0 g, 40.1
mmol) and potassium carbonate (13.8 g, 100 mmol) in ACN (200 mL) add
iodomethane
(5.0 mL, 80.2 mmol). Stir the reaction at 60 C for 15 h. Dilute the reaction
mixture with
water (150 mL) and extract with DCM (3 x 60 mL). Wash the combined organic
layers
with water (50 mL). Dry the organic phase over sodium sulfate, filter, and
concentrate
under reduced pressure to give 10.3 g of the title compound (98%), which is
used in crude
form in Preparation 2. 1H-NMIR (400 MHz, CDC13) 6 7.60 (d, J = 9 Hz, 1H), 7.15
(d, J =
5 Hz, 1H), 3.90 (s, 6H).
Preparation 2
(4-B romo-5-fluoro-2-methoxy -phenyl)methanol
o
OH
Br
To a solution of methyl 4-bromo-5-fluoro-2-methoxy-benzoate (14 g, 53.2 mmol)
and Me0H (30 mL) in THF (300 mL) add sodium borohydride (10.7 g, 272 mmol).
Stir
the reaction at 50 C for 4 h. Concentrate the reaction mixture under reduced
pressure.
Dissolve the residue in Et0Ac (300 mL) and wash with brine (2 x 100 mL). Dry
the
organic phase over sodium sulfate, filter, and concentrate under reduced
pressure. Purify
the residue via silica gel chromatography using a gradient of 0 to 20% Et0Ac
in hexanes
to give 11.5 g of the title compound (92%). 1H-NMR (400 MHz, CDC13) 6 7.14 (d,
J = 8
Hz, 1H), 7.0 (d, J= 6 Hz, 1H), 4.63 (s, 2H), 3.85 (s, 3H).
Preparation 3
1 -B rom o-4-(b romom ethyl)-2-fluoro -5 -methoxy -b enzene
Br
Br
To a solution of (4-bromo-5-fluoro-2-methoxy-phenyl)methanol (11.5 g, 48.9
mmol) in DCM (ZOO mL) add phosphorus tribromide (5.6 mL, 59 mmol). Stir the
reaction
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at RT for 1 h. Quench the reaction with addition of ice water (50 mL) and
basify to pH 7
with saturated aqueous sodium bicarbonate solution. Wash the organic layer
with water
(100 mL). Dry the organic phase over sodium sulfate, filter, and concentrate
under
reduced pressure to give 12.5 g of the title compound (86%), which is used in
crude form
in Preparation 4. 11-1-NMIR (400 MHz, CDC13) 6 7.13 (d, = 8 Hz, 1H), 7.03 (d,
= 6 Hz,
1H), 4.46 (s, 2H), 3.89 (s, 3H).
Preparation 4
2-(4-Bromo-5-fluoro-2-methoxy-phenyl)acetonitrile
o
CN
Br
To a solution of 1-bromo-4-(bromomethyl)-2-fluoro-5-methoxy-benzene (12.5 g,
42.0 mmol) and TMSCN (6.8 mL, 50.5 mmol) in ACN (250 mL) add TBAF solution
(1.0
M in THF, 50 mL, 50 mmol). Stir the reaction at RT for 4 h. Concentrate the
reaction
under reduced pressure. Dissolve the residue in Et0Ac (200 mL) and wash with
saturated
aqueous sodium chloride (2 x 50 mL). Dry the organic phase over sodium
sulfate, filter,
and concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of 0 to 10% Et0Ac in petroleum ether to give 8.0 g of the
title compound
(78%). 11-1-NMIR (400 MHz, CDC13) 6 7.19 (d, J 8 Hz, 1H), 7.04 (d, J = 5 Hz,
1H), 3.86
(s, 3H), 3.64 (s, 2H).
Preparation 5
Ethyl 2-(4-bromo-5-fluoro-2-methoxy-phenyl)acetate
gip CO2Et
Br
To a solution of 2-(4-bromo-5-fluoro-2-methoxy-phenyl)acetonitrile (8.0 g,
32.8
mmol) in ethanol (100 mL) add concentrated sulfuric acid (25 mL). Stir the
reaction at 80
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C for 18 h. Neutralize the reaction to pH 7 with saturated aqueous sodium
bicarbonate
solution. Extract the reaction mixture with DCM (2>< 100 mL). Dry the organic
phase
over sodium sulfate, filter, and concentrate under reduced pressure to give
9.3 g of the
title compound (97%), which is used in crude form in Preparation 6. 1H-NMR
(400 MHz,
CDC13) 6 7.02-6.99 (m, 2H), 4.16 (q, = 7 Hz, 2H), 3_80 (s, 3H), 3.56 (s, 2H),
1.26 (t,
= 7 Hz, 3H).
Preparation 6
Ethyl 2-(4-bromo-5-fluoro-2-hydroxy-phenyl)acetate
OH
(1110 CO2 Et
Br
Cool a solution of ethyl 2-(4-bromo-5-fluoro-2-methoxy-phenyl)acetate (5.0 g,
17.2 mmol) in DCM (100 mL) to -78 C. Add boron tribromide (8.0 mL, 84.8 mmol)
and
stir the reaction at RT for 2 h. Cool the reaction mixture to 0 C and quench
the reaction
with ice water (40 mL). Basify the solution to pH 7 with saturated aqueous
sodium
bicarbonate solution. Wash the organic layer with water (20 mL). Dry the
organic phase
over sodium sulfate, filter, and concentrate under reduced pressure to give
4.0 g of the
title compound (84%), which is used in crude form in Preparations 10, 54 and
60. 1H-
NMR (400 MHz, CDC13) 6 7.69 (s, 1H), 7.14 (d, J = 6 Hz, 1H), 6.90 (d, J= 4 Hz,
1H),
4.22 (q, J= 7 Hz, 2H), 3.61 (s, 2H), 1.31 (t, J= 8 Hz, 3 H).
Preparation 7
Methyl 2-(4-bromo-2-hydroxy-5-methyl-phenyl)acetate
OH
0
Br 0
Prepare the title compound essentially as described in Preparation 6 using
methyl
2-(4-bromo-2-methoxy-5-methyl-phenyl)acetate. ES-MS nilz 259 and 261 (M+H).
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Preparation 8
Methyl 2-[2-hydroxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)phenyliacetate
OH
Mill CO2 M e
B
To a mixture of methyl 2-(4-bromo-2-hydroxy-phenyl)acetate (1.30 g, 5.30
mmol), bis(pinacolato)diboron (1.98 g, 7.72 mmol), KOAc (2.23 g, 22.5 mmol),
and
Pd(dppf)C12 (420 mg, 0.57 mmol) add 1,4-dioxane (24 mL). Stir the reaction
mixture
under a nitrogen atmosphere at 80 C for 60 h. Filter the mixture through a
Celite pad,
and rinse with Et0Ac. Concentrate the filtrate under reduced pressure.
Dissolve the
residue in DCM, adsorb onto silica, and purify via silica gel flash
chromatography using a
gradient of 0 to 55% Et0Ac in hexanes to give 748 mg of the title compound
(48%). ES-
MS nilz 293 (M+H).
Preparation 9
Methyl 2-[2-hydroxy-5-methy1-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
yl)plienyl [acetate
OH
0
0
Prepare the title compound essentially as described in Preparation 8 using
methyl
2-(4-bromo-2-hydroxy-5-methyl-phenyl)acetate. Purify via silica gel flash
chromatography using a gradient of 5 to 80% Et0Ac in hexanes. ES-MS m/z 304 (M-
H).
Preparation 10
Ethyl 2-[4-(5,5-dimethy1-1,3,2-dioxaborinan-2-y1)-5-fluoro-2-
hydroxyphenyl]acetate
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= H
CO2 Et
F
Purge a flask containing ethyl 2-(4-bromo-5-fluoro-2-hydroxy-phenyl)acetate
(2.43 g, 8.16 mmol), bis(neopentyl glycoloato)diboron (2.82 g, 12.2 mmol) and
KOAc
(2.04 g, 20.4 mmol) with nitrogen. Add anhydrous 1,4-dioxane (33 mL) and
sparge with
nitrogen while stirring for 5 min. Add
dichlorobis(tricyclohexylphosphine)palladium(II)
(0.31 g, 0.41 mmol) and sparge with nitrogen while stirring for 5 min. Stir at
90 C for 6
h, then wash down solids using 1,4-dioxane (15 mL) and stir overnight at RT.
Add
diatomaceous earth and dilute with MTBE (0.1 L). Stir for 30 min, filter
through a pad of
Celite and rinse with MTBE (0.1 L). Concentrate the filtrate under reduced
pressure at
50 C. Dissolve the residue in toluene (0.1 L) and concentrate again at 50 C.
Purify the
residue by eluting through a pad of silica gel using a 1:1 mixture of Et0Ac
and heptane.
Concentrate fractions containing the title compound to a final volume of 30 mL
and stir
the resulting slurry for 1 h at ambient temperature. Collect the solid by
filtration, wash
with heptane (0.1 L) and dry under reduced pressure at 50 C for 19 h to
afford 1.77 g of
the title compound (64%) as a pale orange solid. ES-MS rn/z 243 (M+H for
boronic acid).
'1-1-NMR (400 MHz, CDC13) 6 7.28 (s, 1H), 7.27 (d, J = 5.2 Hz, 1H), 6.81 (d, J
= 9.2 Hz,
1H), 4.22 (q, J = 7.2 Hz, 2H), 3.80 (s, 4H), 3.65 (s, 2H), 1.30 (t, J = 7.2
Hz, 3H), 1.05 (s,
6H).
Preparation 11
Ethyl 2-15-fluoro-2-hydroxy-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
yl)phenyllacetate
= H
CO2Et
0
01 F
Prepare the title compound essentially as described in Preparation 10 using
bis(pinacolato)diboron, stirring the mixture at 90 C for 2 h, then 100 C for
18 h. Purify
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the title compound via silica gel flash chromatography using a gradient of 0
to 50%
Et0Ac in cyclohexane, then re-purify via silica gel flash chromatography using
a gradient
of 0 to 40% EtOAc in cyclohexane. ES-MS nilz 325 (M+H).
Preparation 12
(5-Bromo-4-fluoro-2-iodophenyl)methanol
HO
Br
To a solution of 5-bromo-4-fluoro-2-iodo-benzoic acid (6.3 g, 18.2 mmol) in
THF
(55 mL) add borane dimethylsulfide complex (2 M in THF, 27 mL, 54 mmol). Stir
the
reaction mixture at RT for 21 h. Concentrate the reaction mixture and dissolve
the residue
in Et0Ac. Wash the solution with saturated aqueous ammonium chloride. Dry the
organic
phase over magnesium sulfate, filter, and concentrate under reduced pressure.
Purify the
residue via silica gel flash chromatography using a gradient of 10 to 30%
Et0Ac in
heptane to give 5.1 g of the title compound (85%). ES-MS ni/z 313 and 315 (M-
H20).
Preparation 13
Methyl 3,5-difluoro-4-nitro-benzoate
02N 00 CO2Me
Cool a solution of thionyl chloride (37 mL, 74 mmol) in Me0H (110 mL) to -10
C and add 3,5-difluoro-4-nitro-benzonitrile (2.8 g, 15 mmol). Stir at RT for 3
h then
gradually increase the temperature to 65 C over 2 h. Filter the mixture and
concentrate
under reduced pressure. Dissolve the residue in Et0Ac (150 mL). Wash with
saturated
aqueous sodium bicarbonate solution (50 mL) and brine (50 mL). Dry the organic
phase
over sodium sulfate, filter, and concentrate under reduced pressure. Purify
the residue via
silica gel chromatography using 10% Et0Ac in petroleum ether to give 2.24 g of
the title
compound (66%). 11-1-NM_R (400 MHz, CDC13) 6 7.78 (d, 211), 4.0 (s, 3H).
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Preparation 14
Methyl 3-fluoro-4-nitro-5-[[(2S)-oxetan-2-ylm ethyl ]aminoThenzoate
/'""
H N 0
02N CO2Me
Stir a mixture of [(2S)-oxetan-2-yl]methanamine (545 mg, 6.13 mmol, CAS
2091328-57-1), methyl 3,5-difluoro-4-nitro-benzoate (1.4 g, 6.1 mmol), and
potassium
carbonate (1.7 g, 12 mmol) in ACN (14 mL) at 70 C for 16 h. Dilute the
reaction mixture
with water (14 mL) and extract with Et0Ac (3 14 mL). Wash the combined organic
layers with brine (14 mL). Dry the organic phase over sodium sulfate, filter,
and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of 0 to 30% Et0Ac in petroleum ether to give 1.68 g of the
title
compound (79%). ES-MS m/z 285 (M+H).
Preparation I 5
Methyl (S)-3-methoxy-4-nitro-5-((oxetan-2-ylmethyl)amino)benzoate
H N 0
02N CO2Me
0
Prepare the title compound essentially as described in Preparation 14 using
methyl
3-fluoro-5-methoxy-4-nitro-benzoate. Purify the residue via silica gel
chromatography
using a gradient of 5 to 30% Et0Ac in DCM to give the title compound. ES-MS
nilz 296
(M+H).
Preparation 16
Methyl 4-amino-3 -fluoro-5 -[[(2 S)-oxetan-2-ylm ethyl ]aminoThenzoate
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/"¨</N1
HN 0
H 2N CO2Me
To a solution of methyl 3-fluoro-4-nitro-5-[[(2S)-oxetan-2-
ylmethyl]aminoThenzoate (1.68 g, 4.84 mmol) in Me0H (17 mL) add Lindlar
catalyst
containing 5% palladium (600 mg, 0.28 mmol). Stir at RT for 16 h under an
atmosphere
of hydrogen gas. Filter the reaction mixture and concentrate the filtrate
under reduced
pressure to give 1.4 g of the title compound (100%), which is used in crude
form in
Preparation 86 and 91. ES-MS m/z 255 (M+H).
Preparation 17
Methyl 4-amino-3-methoxy-5-[[(25)-oxetan-2-yl]methylaminoThenzoate
HN
H 2 N = CO2Me
0
Prepare the title compound essentially as described in Preparation 16 using
methyl
(S)-3-methoxy-4-nitro-5-((oxetan-2-ylmethyl)amino)benzoate. The title compound
is
used in crude form in Preparations 89, 93, 99, and 100. ES-MS miz 267 (M+H).
Preparation 18
Methyl 3-(((1-ethy1-1H-imidazol-5-y1)methyl)amino)-4-nitrobenzoate
HN Nsi.>
02N LN
"11 CO2Me
To a solution of methyl 3-fluoro-4-nitrobenzoate (300 mg, 1.5 mmol) and TEA
(1.1 mL, 8.1 mmol) in THF (6 mL) and DMF (3 mL), add (1-ethyl-1H-imidazol-5-
yl)methanamine dihydrochloride (339 mg, 1.6 mmol). Stir at 35 C for 2 h, then
50 C for
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16 h. Dilute the crude reaction mixture with water and extract with Et0Ac (3 x
15 mL).
Wash the combined organic layers with water and saturated aqueous NaC1, dry
the
organic phase over magnesium sulfate, filter, and concentrate under reduced
pressure.
Purify the residue via silica gel flash chromatography using a gradient of 0
to 100%
Et0Ac in heptane followed by 5% Me0H in DCM to give 395 mg of the title
compound
(88%). ES-MS nilz 305 (M+H).
Preparation 19
Methyl 4-amino-3-(((1-ethy1-1H-imidazol-5-y1)methyl)amino)benzoate
H
H2N N
14111 10 CO2Me
Stir a solution of iron (193 mg, 3.5 mmol), ammonium chloride (10 mg, 0.19
mmol), and acetic acid (46 mg, 0.77 mmol) in water (3 mL) at 50 C for 15 min.
Add a
solution of methyl 3 -(((1-ethy1-1H-i mi dazol -5 -yl)m ethyl)amino)-4-
nitrobenzoate (117
mg, 0.38 mmol) in DMF (1 mL). Stir at 50 C for 15 min. Quench the reaction
with
aqueous sodium carbonate solution to pH 8 and filter through Celite . Wash the
residue
with water (2 x 20 mL) and back extract the aqueous layer with Et0Ac (2 x 20
mL). Dry
the combined organic phase over magnesium sulfate, filter, and concentrate
under
reduced pressure to give 106 mg of the title compound (100%), which is used in
crude
form in Preparation 95. ES-MS nilz 275 (M+H).
Preparation 20
3-Fluoro-5-methoxy-4-nitrobenzonitrile
02N 401
CN
To a mixture of 5-bromo-1-fluoro-3-methoxy-2-nitrobenzene (700 mg, 2.7 mmol),
zinc cyanide (226 mg, 1.9 mmol), and tetrakis(triphyenylphosphine)palladium(0)
(317
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mg, 0.27 mmol), add DMF (17.8 mL). Stir at 100 C for 1.5 h. Dilute the crude
reaction
mixture with water and extract with Et0Ac. Dry the organic phase over
magnesium
sulfate, filter, and concentrate under reduced pressure. Purify the residue
via silica gel
flash chromatography using a gradient of 0 to 15% Et0Ac in heptane to give 479
mg of
the title compound (89%). ES-MS m/z 197 (M+H).
Preparation 21
(S)-3-Methoxy-4-nitro-5-((oxetan-2-ylmethyl)amino)benzonitrile
H
02N io
CN
Add TEA (1.07 mL, 7.7 mmol) and (S)-oxetan-2-ylmethanamine (235 mg, 2.56
mmoL) to a solution of 3-fluoro-5-methoxy-4-nitrobenzonitrile (457 mg, 2.3
mmol) in
DMF (7 mL). Stir at 35 C overnight. Dilute the crude reaction mixture with
water and
extract with Et0Ac. Dry the organic phase over magnesium sulfate, filter, and
concentrate under reduced pressure. Purify the residue via silica gel flash
chromatography
using a gradient of 0 to 30% Et0Ac in heptane to give 471 mg of the title
compound
(77%). ES-MS m/z 264 (M+H).
Preparation 22
(S)-3-Methoxy-2-nitro-N-(oxetan-2-ylmethyl)-5-(1H-tetrazol-5-y1)aniline
02
H
Add tributyltin azide (1.96 mL, 7.0 mmol) to a solution of (S)-3-methoxy-4-
nitro-
5-((oxetan-2-ylmethyl)amino)benzonitrile (217 mg, 0.82 mmol) in toluene (9.3
mL). Heat
to 150 C in a microwave with stirring for 2.5 h. Filter the crude reaction
mixture through
a plug of 10% w/w KF in silica. Concentrate and triturate the residue with DCM
to give
194 mg of the title compound (62%). ES-MS m/z 307 (M+H).
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Preparation 23
(S)-3-Methoxy-N1-(oxetan-2-ylmethyl)-5-(1H-tetrazol-5-y1)benzene-1,2-diamine
H 2N
11"
Add palladium on carbon (20 mg, 0.009 mmol) to a mixture of (S)-3-methoxy-2-
nitro-N-(oxetan-2-ylmethyl)-5-(1H-tetrazol-5-ypaniline (160 mg, 0.42 mmol) and
Me0H
(3 mL). Stir for 8 h at RT under 4 bar of hydrogen pressure. Filter the crude
reaction
mixture through Celite and concentrate to give 120 mg of the title compound
(52%),
which is used in crude form in Preparation 96. ES-MS nilz 277 (M+H).
Preparation 24
4- [(6-11rom o-2-pyri dyl )oxym ethyl ] -3-i odo-benzonitrile
NC 100 I
0 N Br
To a mixture of 4-(bromomethyl)-3-iodo-benzonitrile (2.88 g, 8.93 mmol), 6-
bromopyridin-2-ol (1.10 g, 6.30 mmol), and silver carbonate (5.1 g, 18.0 mmol)
add 1,4-
dioxane (50 mL). Stir the reaction mixture at 60 C for 15 h. Dilute the
reaction with
Et0Ac (50 mL) and filter through celite. Wash the filtrate with water (2 >< 50
mL) and
saturated aqueous sodium chloride (50 mL). Dry the organic phase over
magnesium
sulfate, filter, and concentrate under reduced pressure. Purify the residue
via silica gel
flash chromatography eluting with a gradient of 5 to 30% Et0Ac in hexanes to
2.8 g of
the title compound (76%) ES-MS m/z 415 and 417 (M+H)
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Preparation 25
Ethyl 4-cyano-3-(3-hydroxypropyl)benzoate
CN
=0 H
CO2 Et
Stir a mixture of nickel (II) bromide (167 mg, 0.76 mmol) and 4,4'-di-tert-
butyl-
2,2'-bipyridine (210 mg, 0.77) in anhydrous 1,4-dioxane (40 mL) while bubbling
with
nitrogen for 15 min at RT. Add ethyl 3-bromo-4-cyanobenzoate (2 g, 7.71 mmol),
3-
bromo- 1-propanol (1.7 mL, 18 mmol) and cobalt (II) phthalocyanine (441 mg,
0.77
mmol). Stir the mixture at RT for 5 min while bubbling with nitrogen. Add
tetrakis(dimethylamino)ethylene (2.5 mL, 11 mmol) and continue stirring the
mixture at
RT for 5 min while bubbling with nitrogen. Seal the vessel and stir the
mixture at 85 C
overnight Cool the mixture to RT, filter through Celite and concentrate under
reduced
pressure. Purify the residue via silica gel flash chromatography using a
gradient of 20 to
50% Et0Ac in cyclohexanes to give the title compound (863 mg, 46%) as a green
solid.
ES-MS m/z 251 (M+NH4-').
Preparation 26
Ethyl 343-[tert-butyl(dimethyl)silyl]oxypropy1]-4-cyano-benzoate
Si'=
CO2 Et
Add tert-butyldimethylchlorosilane (615 mg, 3.96 mmol) and imidazole (298 mg,
433 mmol) to a solution of ethyl 4-cyano-3-(3-hydroxypropyl)benzoate (863 mg,
359
mmol) in DCM (15 mL) at RT. Stir the mixture for 1 h and concentrate under
reduced
pressure. Purify the residue via silica gel flash chromatography using a
gradient of 0% to
50% Et0Ac in cyclohexanes to provide the title compound (1.24 g, 94%) as a
colorless
oil. ES-MS nilz 348 (M+H).
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Preparation 27
243-[tert-Butyl(dimethypsilyfloxypropyl]-4-(hydroxymethyl)benzonitrile
CN
CY Sil<
0 H
Add lithium borohydride in TFIF (2.0 M, 3.9 mL, 7.8 mmol) to a solution of
ethyl
343-[tert-butyl(dimethyl)silyl]oxypropyl]-4-cyano-benzoate (1.24 g, 3.39 mmol)
in
anhydrous TI-IF (9 mL) at 0 C under nitrogen atmosphere. Remove the cooling
bath
after 5 min and stir at RT overnight. Concentrate to remove most of the
reaction solvent
and add citric acid (5%) carefully at 0 C. Extract the aqueous layer with
DCM, combine
organic layers, dry over anhydrous sodium sulfate, filter and concentrate
under reduced
pressure. Purify the residue via silica gel flash chromatography using a
gradient of 10% to
50% Et0Ac in cyclohexanes to provide the title compound (935 mg, 90%) as a
colorless
waxy solid. ES-MS nilz 306 (M+H).
Preparation 28
4-[(6-Bromo-2-pyridyl)oxymethy1]-343-[tert-
butyl(dimethyl)silyl]oxypropyl]benzonitrile
NC Si
\
0 N Br
To a mixture of 4-[(6-bromo-2-pyridyl)oxymethy1]-3-iodo-benzonitrile (2.6 g,
6.3
mmol) and tetrakis(triphenylphosphine)palladium(0) (0.27 g, 0.23 mmol) in 1,4-
dioxane
(50 mL) add bromo[3-ttert-butyl(dimethyl)silylloxypropyl]zinc (0.50 M in THF,
25 mL,
12.5 mmol). Stir at 60 C for 1 h. Dilute the reaction mixture with Et0Ac (100
mL), then
wash with saturated aqueous ammonium chloride solution (100 mL) and brine (100
mL).
Dry the organic phase over sodium sulfate, filter, and concentrate under
reduced pressure.
Purify the residue via silica gel flash chromatography eluting with a gradient
of 5 to 50%
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Et0Ac in hexanes to give 1.07 g of the title compound (37%). ES-MS nilz 461
and 463
(M+H).
Preparation 29
4-[(6-Bromo-2 -pyri dyl)oxymethyl] -3 -(3 -hydroxypropyl)benzonitrile
NC
0 H
O. N Br
To a mixture of 4-[(6-bromo-2-pyridyl)oxymethy1]-3-[3-[tert-
butyhdimethyl)silyl]oxypropyl]benzonitrile (1.32 g, 2.86 mmol) in THE (50 mL)
add
TBAF solution (1.0 M in THE, 2.9 mL, 2.9 mmol). Stir the reaction mixture at
RT for 1
h. Dilute with Et0Ac (50 mL) and wash with saturated aqueous ammonium chloride
solution (50 mL) and brine (50 mL). Dry the organic phase over sodium sulfate,
filter,
and concentrate under reduced pressure. Purify the residue via silica gel
flash
chromatography eluting with a gradient of 5 to 75% Et0Ac in hexanes to give
0.90 g of
the title compound (92%). ES-MS nilz 347 and 349 (M+H).
Preparation 30
4-[(6-Bromo-2-pyridyl)oxymethy11-243-[tert-
butyl(dimethypsilylioxypropylibenzonitrile
CN \
Br
0
j[N
Add DIAD (915 tiL, 4.5 mmol) slowly to a solution of triphenylphosphine (1.21
g,
4.61 mmol) in anhydrous THE (15 mL) at 0 C under nitrogen atmosphere. Stir
the
mixture for 30 min and then add a solution of 243-[tert-
butyl(dimethyl)silyl]oxypropy1]-
4-(hydroxymethyl)benzonitrile (925 mg, 3.03 mmol) in anhydrous THE (6 mL) and
2-
bromo-6-hydroxypyridine (610 mg, 3.33 mmol). Remove the cooling bath, stir at
RT for
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2h, and concentrate the mixture under reduced pressure. Purify the residue via
silica gel
flash chromatography using a gradient of 0 to 30% of Et0Ac in cyclohexanes to
provide
1.12g of the title compound (78%). ES-MS m/z, 461 and 463 (M+H).
Preparation 31
4-[(6-bromo-2-pyridyl)oxymethyl]-2-(3 -hydroxypropyl)b enzonitrile
CN
0 H
0
-
"
Br
Add slowly TBAF solution (1.0 M in THE, 2.7 mL, 2.7 mmol) to a solution of 4-
[(6-bromo-2-pyridyl)oxymethy1]-243-[tert-
butyl(dimethyl)silyl]oxypropylbenzonitrile
(1.12 g, 2.43 mmol) in THE (24 mL) at RT. Stir the mixture for lh. Concentrate
the
reaction mixture and dilute the residue with MTBE and water. Separate the
organic layer
and wash with water and saturated aqueous NaCl, dry over anhydrous Na2SO4,
then filter
and concentrate under reduced pressure. Purify the residue via silica gel
flash
chromatography using a gradient of 25 to 50% of Et0Ac in cyclohexanes to
provide the
title compound (790 mg, 89%) as a white solid. ES-MS in/z 347 and 349 (M+H).
Preparation 32
3-(2-((tert-Butyldimethylsilyl)oxy)ethoxy)-4-formylbenzonitrile
\
NC 0
,0
To a solution of 4-formy1-3-hydroxybenzonitrile (3.0g, 18.8 mmol) in DMF (56
mL), add sodium iodide (1.4 g, 9.3 mmol), potassium carbonate (3.8 g, 38
mmol), and (2-
bromoethoxy)-tert-butyldimethylsilane (6.1 mL, 28 mmol). Stir the mixture at
70 "V for
24 h. Dilute the crude reaction mixture with water and Et0Ac and extract the
aqueous
layer with Et0Ac. Dry the organic phase over magnesium sulfate, filter, and
concentrate
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under reduced pressure. Purify the residue via silica gel flash chromatography
using a
gradient of 0 to 30% Et0Ac in heptane to give 5.68 g of the title compound
(99%). ES-
MS nilz 306 (M+H).
Preparation 33
3-(2-((tert-Butyldimethylsilyl)oxy)ethoxy)-4-(hydroxymethyl)benzonitrile
\ /
NC oC3(Si,i.
0 H
Cool a solution of 3-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-4-
formylbenzonitrile
(450 mg, 1.47 mmol) in Me0H (4.6 mL) to 0 C. Add sodium borohydride (112 mg,
2.96
mmol) and stir the mixture at 0 "V for 15 min. Warm the mixture to RT and stir
for 1 h.
Dilute the mixture with water and adjust to pH 7 with 1 M aqueous HC1
solution, then
extract with Et0Ac. Dry the organic phase over magnesium sulfate, filter, and
concentrate under reduced pressure. Pass the residue through a silica gel pad
to give 350
mg of the title compound (77%). 11-I-NMR (400 MHz, CDC13) 6 7.32 (d, J = 7.7
Hz, 1H),
7.15 (d, J= 7.7 Hz, 1H), 7.02 (d, J= 1.0 Hz, 1H), 4.60 (s, 2H), 4.0 (m, 2H),
3.8 (m, 2H),
3.03 (s, 1H), 0.81 (s, 9H), 0.0 (s, 6H).
Preparation 34
4-(((6-Bromopyridin-2-ypoxy)methyl)-3-(2-((tert-
butyldimethylsilyl)oxy)ethoxy)benzonitrile
\ /
NC C)"0-SiK
=
0 N Br
To a solution of 3-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-4-
(hydroxymethyl)benzonitrile (350 mg, 1.14 mmol), 6-bromopyridin-2-ol (1.4 g,
8.0
mmol), and triphenylphosphine (2.35 g, 8.96 mmol) in THF (50 mL) add DIAD
(1.76
mL, 8.94 mmol). Stir the mixture at 50 C for 4 h and then concentrate the
crude reaction
mixture. Dilute the residue with Et0Ac, then wash with water (3x) and brine.
Dry the
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organic phase over magnesium sulfate, filter, and concentrate under reduced
pressure.
Purify the residue via silica gel flash chromatography using a gradient of 0
to 20% Et0Ac
in heptane to give 247 mg of the title compound (47%). ES-MS m/z 463 and 465
(M+H).
Preparation 35
4-4(6-Bromopyridin-2-yl)oxy)methyl)-3-(2-hydroxyethoxy)benzonitrile
NC
o H
0 N Br
Prepare the title compound essentially as described in Preparation 31 using 4-
(((6-
bromopyridin-2-yl)oxy)methyl)-3-(2-((tert-
butyldimethylsily1)oxy)ethoxy)benzonitrile,
stirring the reaction at RT for 2 h. Purify the title compound via silica gel
flash
chromatography using a gradient of 0 to 20% Et0Ac in DCM. ES-MS m/z 349 and
351
(M+H).
Preparation 36
Ethyl (E)-3-[5-chloro-2-(hydroxymethyl)phenyl]prop-2-enoate
0
CI
0 Et
0 H
To a mixture of (2-bromo-4-chloro-phenyl)methanol (17.5 g, 79.0 mmol),
tetrabutylammonium chloride (26.1 g, 93.9 mmol), potassium carbonate (16.7 g,
121
mmol), and palladium acetate (1.47 g, 6.55 mmol) in DMF (400 mL) add ethyl
acrylate
(10.3 mL, 94.8 mmol). Stir the reaction mixture under a nitrogen atmosphere at
90 C for
6 h. Filter the reaction mixture through a Celite pad, then rinse with Et0Ac
(200 mL).
Dilute the filtrate with EtOAc (200 mL) and wash with water (800 mL). Back-
extract the
aqueous layer with Ft0Ac (250 mL). Dry the combined organic phase over
magnesium
sulfate, filter, and concentrate under reduced pressure. Dissolve the residue
in DCM,
adsorb onto silica, and purify via silica gel chromatography using a gradient
of 15 to 40%
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Et0Ac in hexanes to give 6.14 g of the title compound (32%). ES-MS nilz 258
(M+NE-14)_
Preparation 37
Ethyl 345-chloro-2-(hydroxymethyl)phenyl]propanoate
0
CI
0 Et
0 H
To a mixture of sulfided platinum on carbon (5%, 220 mg, 0.056 mmol) in Et0Ac
(20 mL) add ethyl (E)-3-[5-chloro-2-(hydroxymethyl)phenyl]prop-2-enoate (2.21
g, 9.18
mmol) in Et0Ac (30 mL). Shake in a Parr shaker at RT at 40 psi of hydrogen gas
pressure for 1 h. Filter through a pad of Celite and concentrate the filtrate
under reduced
pressure to provide 1_91 g of the title compound (86%), which is used in crude
form in
Preparation 38. ES-MS nilz 225 (M-H20).
Preparation 38
Ethyl 3-[2-(bromomethyl)-5-chloro-phenyl]propanoate
0
CI
0 Et
Br
To a mixture of ethyl 3[5-chloro-2-(hydroxymethyl)phenyl]propanoate (1.90 g,
7.80 mmol) in diethyl ether (40 mL) dropwise add phosphorus tribromide (0.80
mL, 8.5
mmol). Stir under a nitrogen atmosphere at RT for 1 h. Quench with slow,
dropwise
addition of saturated aqueous sodium bicarbonate solution (5 mT,). Separate
the layers
and extract the aqueous layer with diethyl ether (5 mL). Dry the combined
organic phase
over magnesium sulfate, filter, and concentrate under reduced pressure to
provide 2.40 g
of the title compound (100%), which is used in crude form in Preparation 45.
ES-MS miz
322 and 324 (M-Fl\THI ).
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Preparation 39
4-[(1,3-Dioxoisoindolin-2-yl)methyl]-3-iodo-benzonitrile
NC
0 0
To a solution of 4-(bromomethyl)-3-iodobenzonitrile (10 g, 30,13 mmol) in DMF
(100mL), add potassium phthalimide (6,14 g, 33,14 mmol). Heat the reaction
mixture at
80 C for 2 h and then stir at RT for 16 h. Remove solvent and triturate the
solid residue
in 500 mL of water for 30 min. Filter the white solid, wash with water and dry
the solid
under vacuum at 45 C for 20 h, to provide the title compound as a white solid
(11.5 g,
88%). ES-MS nilz 405 (M-F0H-).
Preparation 40
4-1(1,3 -Dioxoi soindolin-2-yl)methy11-3 -(3 -hydroxyprop -1 -
ynyl)benzonitrile
0 N H
C
0 0
To a suspension of 4-[(1,3-dioxoisoindolin-2-ypmethyl]-3-iodo-benzonitrile
(5.0
g, 11.7 mmol) in THE (50 mL) and TEA (50 mL), add
bis(triphenylphosphine)palladium
dichloride (0.33 g, 0.47 mmol), cuprous iodide (0.18 g, 0.94 mmol) and
propargyl alcohol
(2.05 mL, 35.2 mmol). Heat reaction mixture at 40 C for 3 h. Cool to RT,
dilute with
Et0Ac (50 mL) and water (50 mL) and filter the mixture through Celite .
Separate
phases and extract aqueous phase with more Et0Ac (2 x 50 mL). Combine
organics, dry
over magnesium sulfate, filter, and concentrate under reduced pressure. Purify
the residue
via silica gel chromatography using a gradient of 30% of Et0Ac in cyclohexane
to 100%
Et0Ac to give 3.2g (79%) of the title compound as a cream solid. ES-MS m/z 315
(M-H).
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Preparation 41
4-[(1,3-Dioxoisoindolin-2-yl)methyl]-3-(3-hydroxypropyl)benzonitrile
N C
0 H
0 0
In a 250 mL Buchi Mini cl ave reactor, to a suspension of 4-[(1,3-
dioxoisoindolin-2-yl)methy1]-3-(3-hydroxyprop-1-ynyl)benzonitrile (3 g, 9.48
mmol) in
Me0H (60 mL), add 1, l'-bis(di-i-propylphosphino)ferrocene(1,5-
cyclooctadiene)rhodium(I) tetrafluoroborate (0.34 g, 0.47 mmol). Charge
reactor with 90
psi of hydrogen and heat at 50 C for 2 h. Cool reaction mixture to RT,
evaporate solvents
and purify the residue via silica plug using Et0Ac as solvent, to provide 2.4g
(75%) of
the title compound as a light cream solid. ES-MS m/z 321 (M+H).
Preparation 42
4-(Aminomethyl)-3-(3-hydroxypropyl)benzonitrile
N C
0 H
N H 2
To a suspension of 4-[(1,3-dioxoisoindolin-2-yl)methyl]-3-(3-
hydroxypropyl)benzonitrile (2.4 g, 7.49 mmol) in Me0H (45 mL), add hydrazine
monohydrate (1.90 mL, 37.6 mmol) and stir at 60 C for 20 h. Cool reaction
mixture to RT
and filter the solid. Evaporate filtrate liquids, dilute the residue with
water (30 mL) and
extract with DCM/Me0H 9:1 (3 x 20 mL). Combine organics, dry over magnesium
sulfate, filter, and concentrate under reduce pressure to give the title
compound as a
reddish oil (880 mg, 49%). ES-MS nvz 191 (M+H).
Preparation 43
Methyl 2-(((5-bromo-4-fluoro-2-iodobenzyl)oxy)methyl)-4-cyanobenzoate
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NC
0110 0
02Me ISM
Br
To a solution of (5-bromo-4-fluoro-2-iodophenyl)methanol (1.5 g, 4.5 mmol) in
THF (24 mL) at 0 C add sodium hydride (60% in mineral oil, 362 mg, 9.1 mmol).
Stir at
0 C for 30 min. Add methyl 2-(bromomethyl)-4-cyano-benzoate (2.3 g, 9.1 mmol)
and
stir at RT for 1 h. Dilute the reaction with Et0Ac and wash the organic layer
with water
and saturated aqueous NaCl. Back extract the aqueous layer with Et0Ac. Dry the
organic
phase over magnesium sulfate, filter, and concentrate under reduced pressure.
Purify the
residue via silica gel flash chromatography using DCM to give 2.5 g of the
title
compound (67%) which is 61% pure, and used without further purification in
Preparation
44. ES-MS nilz 504 and 506 (M+H).
Preparation 44
3-(((5-Bromo-4-fluoro-2-iodobenzyl)oxy)methyl)-4-(hydroxymethyl)benzonitrile
NC,
0
1101
0 H Br
Prepare the title compound essentially as described in Preparation 27 using
methyl
2-(((5-bromo-4-fluoro-2-iodobenzyl)oxy)methyl)-4-cyanobenzoate (Preparation
43) and a
10:1 mixture of THF:Me0H as reaction solvent. After stirring the reaction at
RT for 20
h, add an additional portion of lithium borohydride (0.5 equivalents) and stir
at RT for 2
h. Dilute the reaction mixture with Et0Ac and wash the organic layer with
water and
saturated aqueous NaC1 Dry the organic phase over magnesium sulfate, filter,
and
concentrate under reduced pressure. Purify the residue via silica gel flash
chromatography
using a gradient of 0 to 50% Et0Ac in heptane to give the title compound. ES-
MS nilz
476 and 478 (M+H).
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Preparation 45
Ethyl 342-[(6-bromo-2-pyridyl)oxymethyl]-5-chloro-phenyl]propanoate
CI CO2Et
0 N Br
To a mixture of ethyl 3[2-(bromomethyl)-5-chloro-phenyl]propanoate (2.40 g,
7.85 mmol), 6-bromopyridin-2-ol (1.98 g, 11.4 mmol), and silver carbonate
(4.34 g, 15.7
mmol) add 1,4-dioxane (40 mL). Stir the reaction mixture at 40 C for 15 h.
Filter through
a pad of Celite and concentrate the filtrate under reduced pressure. Dissolve
the residue
in DCM, adsorb onto silica, and purify via silica gel chromatography using a
gradient of 0
to 40% Et0Ac in hexanes to give 1.16 g of the title compound (37%). ES-MS in/z
398,
400 and 402 (M+H).
Preparation 46
342-1(6-Bromo-2-pyridyl)oxymethy1]-5-chloro-phenyl]propan-1-ol
CI
0 H
0 N Br
To a mixture of ethyl 342-[(6-bromo-2-pyridyl)oxymethy1]-5-chloro-
phenyl]propanoate (1.16 g, 2.90 mmol) in TI-IF (12 mL) dropwise add lithium
borohydri de (2.0 M in THF, 3.2 mL, 6.4 mmol). Stir the reaction mixture at RT
for 6 h.
Add an additional portion of lithium borohydride (2.0 M in THF, 2.0 mL, 4.0
mmol). Stir
the reaction mixture at RT for an additional 17 h. Quench the reaction with
dropwi se
addition of water. Dilute the mixture with Et0Ac (50 mL) and wash with water
(40 mL).
Back extract the aqueous layer with Et0Ac (25 mL). Dry the combined organic
phase
over magnesium sulfate, filter, and concentrate under reduced pressure.
Dissolve the
residue in DCM, adsorb onto silica, and purify via silica gel flash
chromatography eluting
with a gradient of 0 to 55% Et0Ac in hexanes to give 461 mg of the title
compound
(35%). ES-MS nilz 356, 358 and 360 (M+H).
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Preparation 47
4-[[(6-Bromo-2-pyridyl)amino]methy1]-3-(3-hydroxypropyl)benzonitrile
NC
0 H
HN N Br
To a solution of 4-(aminomethyl)-3-(3-hydroxypropyl)benzonitrile (800 mg, 3.36
mmol) in DMSO (16 mL), add 2-bromo-6-fluoropyridine (610 mg, 3.36 mmol) and
DIPEA (1.17 mL, 6.72 mmol) and stir at 100 C for 18 h. Cool reaction mixture
to RT,
dilute with water (40 mL), and extract with Et0Ac (3 x 20 mL). Combine
organics, dry
over magnesium sulfate, filter, and concentrate under reduce pressure. Purify
the residue
via silica gel chromatography using a gradient of 30% of Et0Ac in cyclohexanes
to 100%
Et0Ac to provide the title compound as a thick brown oil (400 mg, 33%). ES-MS
111/1Z 346
and 348 (M+H).
Preparation 48
4-[(3-Bromophenoxy)m ethy1]-3-iodo-benzonitrile
NC I
0 Br
To a mixture of 4-(bromomethyl)-3-iodo-benzonitrile (2.0 g, 6.2 mmol), 3-
bromophenol (1.10 g, 6.4 mmol), and potassium carbonate (2.6 g, 19.0 mmol) add
1,4-
dioxane (20 mL). Stir the reaction mixture at RT for 15 h. Dilute the reaction
with Et0Ac
(100 mL) and filter through Celite . Wash the filtrate with water (2>< 50 mL)
and
saturated aqueous NaC1 (50 mL) Dry the organic phase over sodium sulfate,
filter, and
concentrate under reduced pressure. Purify the residue via silica gel flash
chromatography
eluting with a gradient of 0 to 30% Et0Ac in hexanes to afford 2.4 g of the
title
compound (93%). 1E1 NMR (400.13 MHz, CDC13) 6 8.16 (d,
1.5 Hz, 1H), 7.71 (dd, J-
1.5, 8.0 Hz, 1H), 7.64 (d, J= 8.0 Hz, 1H), 7.23-7.17 (m, 3H), 6.92 (dt, J=
7.5, 2.1 Hz,
1H), 5.05 (s, 2H).
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Preparation 49
44(3-Bromophenoxy)methy1]-343-[tert-
butyl(dimethyl)silyl]oxypropylibenzonitrile
CN
0 k -s,
11101 Br
Prepare the title compound essentially as described in Preparation 28 using 4-
[(3-
bromophenoxy)methy1]-3-iodo-benzonitrile. ES-MS nilz 460 and 462 (M+H).
Preparation 50
4-[(3-Bromophenoxy)methy1]-3-(3-hydroxypropyl)benzonitrile
CN
0 H
=
Br
Prepare the title compound essentially as described in Preparation 29 using 4-
[(3-
bromophenoxy)methy1]-343-[tert-butyl(dimethypsilyl]oxypropyl]benzonitrile. ES-
MS
in/z 344 and 346 (M-H).
Preparation 51
2-Bromo-6-[[2-iodo-4-(trifluoromethyl)phenyl]methoxy]pyridine
F3C
0 N Br
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To a mixture of [2-iodo-4-(trifluoromethyl)phenyl]methanol (2.0 g, 6.6 mmol),
2-
bromo-6-fluoro-pyridine (1.2 g, 6.6 mmol), and 1,4-dioxane (25 mL) add
potassium tert-
butoxide (0.98 g, 8.6 mmol). Stir the reaction mixture at 50 C for 2 h.
Dilute the reaction
with Et0Ac (100 mL) and filter through Celite Wash the filtrate with water (2
x 50 mL)
and saturated aqueous sodium chloride (50 mL). Dry the organic phase over
sodium
sulfate, filter, and concentrate under reduced pressure. Purify the residue
via silica gel
flash chromatography eluting with a gradient of 10 to 50% Et0Ac in hexanes to
2.0 g of
the title compound (66%). ES-MS nilz 458 and 460 (M+H).
Preparation 52
342-[(6-Bromo-2-pyridypoxymethy1]-5-(trifluoromethyl)phenyl]propoxy-tert-butyl-
dimethyl-silane
F3C Si
\
0 N Br
Prepare the title compound essentially as described in Preparation 28 using 2-
bromo-64[2-iodo-4-(trifluoromethyl)phenyl]methoxy]pyridine. Purify the title
compound
via silica gel flash chromatography eluting with a gradient of 0 to 10% Et0Ac
in hexanes.
Use directly in Preparation 53 without further characterization.
Preparation 53
342-[(6-Bromo-2-pyridyl)oxymethyl]-5-(trifluoromethyl)phenyl]propan-1-ol
F3C 0 H
0 N Br
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Prepare the title compound essentially as described in Preparation 29 using
342-
[(6-bromo-2-pyridyl)oxymethy1]-5-(trifluoromethyl)phenyl]propoxy-tert-butyl-
dimethyl-
sil ane. ES-MS in/z 390 and 392 (M+H).
Preparation 54
Ethyl 244-bromo-24342-[(6-bromo-2-pyridyl)oxymethyl]-5-cyano-phenyl]propoxy]-5-
fluoro-phenyl]acetate
CO2Et
NC
0 1411 Br
0 Br
To a mixture of 4-[(6-bromo-2-pyri dyl)oxym ethyl] -3-(3-
hydroxypropyl)benzonitrile (550 mg, 1.58 mmol) and ethyl 2-(4-bromo-5-fluoro-2-
hydroxy-phenyl)acetate (0.40 g, 1.4 mmol) in THF (10 mL) add tri-n-
butylphosphine
(0.90 mL, 4.0 mmol). Dropwise add a solution of DEAD (40% solution in toluene,
1.1
mL, 2.8 mmol) in DCM (1.1 mL). Stir the reaction mixture at RT for 15 h.
Quench the
reaction with Me0H (5 mL) and concentrate under reduced pressure. Purify the
residue
via silica gel flash chromatography eluting with a gradient of 5 to 40% Et0Ac
in hexanes
to give 1.1 g of the title compound (92%). ES-MS nilz 605, 607, and 609 (M+H).
Preparation 55
Methyl 2-[2-[3-[2-[(6-bromo-2-pyridyl)oxymethy1]-5-chloro-phenyl]propoxy]-4-
(4,4,5,5-
tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate
CO2Me
CI
0 1E?-17t.
0 Ny Br
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To a mixture of 3-[2-[(6-bromo-2-pyridyl)oxymethy11-5-chloro-phenyllpropan-1-
01 (420 mg, 1.18 mmol), methyl 2-[2-hydroxy-4-(4,4,5,5-tetramethy1-1,3,2-
dioxaborolan-
2-yl)phenyl]acetate (422 mg, 1 45 mmol), and triphenylphosphine (460 mg, 1.75
mmol)
in TETT (8 mL) dropwise add DIAD (0.35 mL, 1.80 mmol). Stir the reaction
mixture at
RT for 1.5 h. Adsorb the mixture onto silica and purify via silica gel flash
chromatography eluting with a gradient of 0 to 30% Et0Ac in hexanes to give
306 mg of
the title compound (41%). ES-MS nilz 630 and 632 (M+H).
Preparation 56
Methyl 2-[2-[3-[2-[[(6-bromopyridin-2-yl)oxy]methy1]-5-cyanophenyl]propoxy]-4-
(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate
CO2Me
NC
0 I. Bt
0 N Br
To a mixture of 4-[(6-bromo-2-pyridyl)oxymethy1]-3-(3-
hydroxypropyl)benzonitrile (14.5 g, 41.9 mmol) and methyl 2-[2-hydroxy-4-
(4,4,5,5-
tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate (18.7 g, 63.9 mmol) in THE
(250 mL)
add tri-n-butylphosphine (21 mL, 84 mmol). Cool the solution in an ice bath
and
dropwise add DIAD (17 mL, 86 mmol). Stir the reaction mixture at 45 C for 14
hand
then concentrate under reduced pressure. Suspend the residue in Et0Ac (100 mL)
and
collect the solid by filtration. Wash the solid with Et0Ac (3 >< 25 mL) to
give 19.7 g of
the title compound (76%). ES-MS nilz 621, 623 (M+H).
Preparation 57
Ethyl 2424342-[[(6-bromo-2-pyridyl)aminolmethy11-5-cyano-phenyl]propoxy]-4-
(5,5-
dimethyl-1,3,2-dioxaborinan-2-y1)-5-fluoro-phenyl]acetate
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CO2Et
NC 0
0
=
HN N Br
To a solution of 4-[[(6-bromo-2-pyridyl)amino]methy1]-3-(3-
hydroxypropyl)benzonitrile (380 mg, L04 mmol) and ethyl 244-(5,5-dimethy1-
1,3,2-
dioxaborinan-2-y1)-5-fluoro-2-hydroxyphenyl]acetate (0.38 g, 1.15 mmol) in THF
(5.2
mL) at 4 C, add triphenylphosphine (0.30g, 1.15 mmol) and di-tertbutyl
azodicarboxylate
(0.27 g, 1.15 mmol). Stir at RT for 22 h, concentrate the reaction mixture and
purify the
residue via silica gel chromatography using a gradient of 10 to 70% Et0Ac in
cyclohexanes to get the title compound as a pale brown solid (500 mg, 60%). ES-
MS nilz
570 and 572 (M+H for boronic acid).
Preparation 58
Methyl 2424342-[(6-bromo-2-pyridyl)oxymethyl]-5-cyano-phenyl]propoxy]-5-methyl-
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]acetate
Me02C
NC 0
s'o _________________________________________________________
0 N Br
Prepare the title compound essentially as described in Preparation 55 using 4-
[(6-
bromo-2-pyridyl)oxymethy1]-3-(3-hydroxypropyl)benzonitrile and methyl 2-[2-
hydroxy-
5-methy1-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)phenyliacetate. Use
tri-n-
butylphosphine instead of triphenylphosphine, DEAD instead of DIAD, and use
dioxane
as solvent instead of THF. Purify via silica gel flash chromatography using a
gradient of
80-100% DCM in hexanes ES-MS nilz 635 and 637 (M+H)
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Preparation 59
Ethyl 2424345-[(6-bromo-2-pyridyl)oxymethyl]-2-cyano-phenyl]propoxy]-5-fluoro-
4-
(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate
CO Et
CN
0 110 B-76
N
Br
Add slowly a solution of di-tertbutyl azodicarboxylate (390 mg, 1.7 mmol) in
anhydrous '11414 (2 mL) to a solution of triphenylphosphine (435mg, 1.66mmo1),
44(6-
bromo-2-pyridyl)oxymethy1]-2-(3-hydroxypropyl)benzonitrile (400 mg, 1.09 mmol)
and
ethyl 245-fluoro-2-hydroxy-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
yl)phenyllacetate (480 mg, purity 83%, 1.23 mmol) in anhydrous THF (8 mL) at
RT. Stir
the mixture at RT for 1 h. Add triphenylphosphine (170mg, 0.65mmo1), and after
5min
add slowly a solution of di-tertbutyl azodicarboxylate (157 mg, 0.65 mmol) in
anhydrous
THF (2 m1). Concentrate the reaction mixture and purify the residue via silica
gel flash
chromatography using a gradient of 10 to 50 % Et0Ac in cyclohexanes to give
the title
compound as a brown waxy solid (689 mg, purity 90%, 86%). ES-MS m/z 653 and
655
(M+H).
Preparation 60
Ethyl 2-(4-bromo-2-(2-(2-(((6-bromopyridin-2-yl)oxy)methyl)-5-
cyanophenoxy)ethoxy)-
5-fluorophenyl)acetate
CO2Et
NC
ei 0,0 el Br
0 N Br
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To a solution of 4-(((6-bromopyridin-2-yl)oxy)methyl)-3-(2-
hydroxyethoxy)benzonitrile (640 mg, 1.83 mmol), ethyl 2-(4-bromo-5-fluoro-2-
hydroxy-
phenyl)acetate (506 mg, 1.83 mmoL), and TMAD (671 mg, 3.70 mmol) in TI-IF (9
mL)
add tri-N-butylphosphine (0.91 mL, 3.65 mmol) Stir at 35 C for 2 h. Quench
the
reaction with Me0H and concentrate the crude mixture. Purify the residue via
silica gel
flash chromatography using a gradient of 0 to 30% Et0Ac in heptane to give 876
mg of
the title compound (79%). ES-MS nilz 607, 609, and 611 (M+H).
Preparation 61
3#(5-Bromo-4-fluoro-2-iodobenzyl)oxy)methyl)-4-(((6-bromopyridin-2-
ypoxy)methyl)benzonitrile
NC
0
0 Br
Ny
Br
Prepare the title compound essentially as described in Preparation 34 using 3-
(((5-
bromo-4-fluoro-2-iodobenzyl)oxy)methyl)-4-(hydroxymethyl)benzonitrile. Purify
the title
compound via silica gel flash chromatography using a gradient of 0 to 10%
Et0Ac in
heptane. ES-MS nvz 631, 633, and 635 (M+H).
Preparation 62
Ethyl 2-(4-bromo-2-(((2-(((6-bromopyridin-2-yl)oxy)methyl)-5-
cyanobenzyl)oxy)methyl)-5-fluorophenypacetate
CO2 Ft
NC
* 0
Br
Nyl
Br
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To a solution of 3-(((5-bromo-4-fluoro-2-iodobenzyl)oxy)methy1)-4-(((6-
bromopyridin-2-yl)oxy)methyl)benzonitrile (2.69 mg, 4.3 mmol) in ACN (75 mL),
add
bis(triphenylphosphine)palladium (II) dichloride (305 mg, 0.43 mmol), TEA
(1.48 mL,
10.6 mmol), and formic acid (0.24 mL, 6.4 mmol). Stir the reaction at 70 C
and add a
solution of ethyl diazoacetate (2 M in DCM, 8.5 mL, 17 mmol) in ACN (25 mL)
over 10
min. Stir at 70 C for 2 h. Add second portions (half the amounts of initial
addition) of all
reagents and heat for a further 1.5 h. Dilute the reaction mixture with Et0Ac
and wash the
organic layer with water. Dry the organic phase over magnesium sulfate,
filter, and
concentrate under reduced pressure. Purify the residue via silica gel flash
chromatography
using a gradient of 0 to 10% Et0Ac in heptane to give 898 mg of the title
compound
(36%). ES-MS nilz 591, 593, and 595 (M+H).
Preparation 63
Methyl 24243-12-[(3-bromophenoxy)methy1]-5-cyano-phenyl]propoxy]-5-methy1-4-
(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate
Me02C
NC 4111
0
0 ____________________________________________________________
0 io Br
Prepare the title compound essentially as described in Preparation 54 using 4-
[(3-
bromophenoxy)methy1]-3-(3-hydroxypropyl)benzonitrile and methyl 2-[2-hydroxy-5-
methy1-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)phenyl]acetate, using
1,4-dioxane
as reaction solvent and adding DEAD as a 40% solution in toluene. Purify the
title
compound via silica gel flash chromatography eluting with a gradient of 80 to
100%
DCM in hexanes. ES-MS nilz 651 and 653 (M+NH4+).
Preparation 64
Methyl 24213-12-[(6-bromo-2-pyridyl)oxymethy1]-5-
(trifluoromethyl)phenyl]propoxy1-
5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-y1)phenyl]acetate
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Me02C
F3C 0 I. y
0
O. N Br
Prepare the title compound essentially as described in Preparation 54 using
342-
[(6-bromo-2-pyridyl)oxymethy1]-5-(trifluoromethyl)phenyl]propan-1-ol and
methyl 2-[2-
hydroxy-5-methy1-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
y1)phenyl]acetate, adding
DEAD to the reaction as a 40% solution in toluene. Purify the title compound
via silica
gel flash chromatography eluting with a gradient of 85 to 100 % DCM in
hexanes. ES-
MS 117//Z 678 and 680 (M+H).
Preparation 65
Ethyl 2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
NC
0
CO2Et
0 N
To a solution of ethyl 244-bromo-24342-[(6-bromo-2-pyridyl)oxymethyl]-5-
cyano-phenyl]propoxy]-5-fluoro-phenyl]acetate (1.1 g, 0.92 mmol) in 1,4-
dioxane (40
mL) add hexamethylditin (0.71 g, 2.2 mmol). Add
tetrakis(triphenylphosphine)palladium(0) (0.20 g, 0.20 mmol). Stir the
reaction mixture at
90 C for 60 h. Concentrate under reduced pressure and purify the residue via
silica gel
flash chromatography eluting with a gradient of 5 to 45% Et0Ac in hexanes to
give 303
mg of the title compound (47%). ES-MS m/z 447 (M+H).
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Preparation 66
Methyl 2-(54-chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetate
CI
0
CO2Me
0 N
I
To a mixture of methyl 2-[2-[342-[(6-bromo-2-pyridy1)oxymethy1]-5-chloro-
phenyl]propoxy]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]acetate
(306 mg,
0.49 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-
biphenyl)(2'-
amino-1,1'-biphenyl-2-y1) palladium(II) (XPhos Gen 2, 28.9 mg, 0.036 mmol),
and
potassium phosphate (420 mg, 1.94 mmol) add THF (48 mL) and water (5.4 mL).
Stir the
reaction mixture under a nitrogen atmosphere at 40 C for 15 h. Concentrate
the mixture
under reduced pressure. Dissolve the residue in DCM, adsorb onto Celite , and
purify via
silica gel flash chromatography eluting with a gradient of 0 to 40% Et0Ac in
hexanes to
give 152 mg of the title compound (74%). ES-MS in/ 424 and 426 (M+H).
Preparation 67
Methyl 2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetate
NC 0
CO2Me
0
I
To a mixture of methyl 2424342-[[(6-bromopyridin-2-yl)oxy]methyl]-5-
cyanophenyllpropoxy1-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
yl)phenyllacetate
(19.7 g, 31.7 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-
biphenyl)(2'-amino-1,1'-biphenyl-2-y1) palladium(II) (XPhos Gen 2, 1.3 g, 1.6
mmol),
and potassium phosphate (28.5 g, 132 mmol) add THF (500 mL) and water (52 mL).
Stir
the reaction mixture under a nitrogen atmosphere at 45 C for 2 h 15 min.
Dilute the
reaction with Et0Ac (200 mL) and wash with half-saturated brine (400 mL). Dry
the
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organic phase over MgSO4, filter, and concentrate under reduced pressure.
Suspend the
residue in Et0Ac (100 mL) and collect the solid by filtration. Wash the solid
with Et0Ac
(3 x 30 mL) to give 10.7 g of the title compound (82%). ES-MS nilz 415 (M+H)
Preparation 68
Ethyl 2-(54-cyano-16-fluoro-9-oxa-3-aza-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
NC 0
CO2 Et
H N N
To a solution of ethyl 24243-[2-[[(6-bromo-2-pyridyl)amino]methyl]-5-cyano-
phenyl]propoxy]-4-(5,5-dimethy1-1,3,2-dioxaborinan-2-y1)-5-fluoro-
phenyl]acetate (460
mg, 0.57 mmol) in 1,4-dioxane (11.5 mL), add PdC12(dtbpf) (77 mg, 0.11 mmol)
and 1M
aqueous potassium phosphate solution (1.73 mL, 1.73 mmol). Stir at 70 C for 2
h, cool to
RT and dilute the reaction mixture with saturated ammonium chloride solution
(15 mL)
and Et0Ac ( I 0 mL). Separate phases, extract aqueous phase with Et0Ac (2>< 5
mL).
Combine organics, dry over magnesium sulfate, filter, and concentrate under
reduced
pressure. Purify the residue via silica gel chromatography using 30% of Et0Ac
in
cyclohexanes as eluent system to provide the title compound (125 mg, 49%) as a
light-
yellow solid. ES-MS nilz 446 (M+H).
Preparation 69
Methyl 2-(54-cyano-16-methyl -3,9-di oxa-2(2,6)-pyri dina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
NC
0
CO2Me
0 N
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Prepare the title compound essentially as described in Preparation 66 using
methyl
2424342-[(6-bromo-2-pyridyl)oxymethy1]-5-cyano-phenyl]propoxy]-5-methyl-4-
(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenydacetate. Purify via silica
gel flash
chromatography using a gradient of 0 to 20% Et0Ac in DCM. ES-MS in/z 429
(M+H).
Preparation 70
Ethyl 2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)acetate
CN
0
CO2Et
0 N
To a solution of ethyl 2424345-[(6-bromo-2-pyridyl)oxymethyl]-2-cyano-
phenyl]propoxy]-5-fluoro-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
yl)phenyllacetate
(650 mg, 90% purity, 0.89 mmol) in 1,4-dioxane (30 mL), add PdC12(dtbpf) (119
mg,
0.18 mmol) and 1M aqueous potassium phosphate solution (2.7 mL, 2.7 mmol).
Stir the
mixture at 50 C for 15 min under nitrogen atmosphere, then cool to RT and
dilute with
DCM. Dry over anhydrous Na2SO4, filter, and concentrate under reduced
pressure.
Purify the residue via silica gel flash chromatography using a gradient of 10
to 30%
Et0Ac in cyclohexanes to provide the title compound (150 mg, 38%) as a very
pale
brown solid. ES-MS m/z 447 (M+H).
Preparation 71
Ethyl 2-(54-cyano-16-fluoro-3,6,9-trioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
NC
CO2Et
0 N.,
F
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To a solution of ethyl 2-(4-bromo-2-(2-(2-(((6-bromopyridin-2-ypoxy)methyl)-5-
cyanophenoxy)ethoxy)-5-fluorophenypacetate (400 mg, 0.66 mmol), KOAc (0.2 g,
2.0
mmol), bis(pinacolato)diboron (187 mg, 0.72 mmol) in 1,4-dioxane (2.2 mL) add
Pd(dppf)C12-DCM complex (27 mg, 0.032 mmol). Stir the mixture at 85 C for 1
h, then
dilute the with water and extract three times with Et0Ac. Dry the organic
phase over
magnesium sulfate, filter, and concentrate under reduced pressure. To the
residue, add
THF (66 mL), potassium phosphate tribasic (0.6 g, 3.0 mmol), water (7.3 mL),
and a
premade solution of palladium (II) chloride (6.0 mg, 0.033 mmol) and 2-
dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (32 mg, 0.066 mmol) in THF
(1
mL). Stir at 45 C for 16 h. Dilute the crude mixture with water and extract
three times
with Et0Ac (3x). Dry the organic phase over magnesium sulfate, filter, and
concentrate
under reduced pressure. Purify the residue via silica gel flash chromatography
using a
gradient of 0 to 30% Et0Ac in heptane to give 94 mg of the title compound
(32%). ES-
MS nilz 449 (M+H).
Preparation 72
Ethyl 2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-ypacetate
NC
010 0
CO2 Et
0 N
F
Prepare the title compound essentially as described in Preparation 65 using
ethyl
2-(4-bromo-2-(((2-(((6-bromopyridin-2-yl)oxy)methyl)-5-cyanobenzyl)oxy)methyl)-
5-
fluorophenyl)acetate, using 1.1 eq of hexamethylditin and with Pd(dppf)C12-DCM
as
catalyst, and stirring the reaction at 100 "V for 3.5 h. Dilute the crude
reaction mixture
with water and extract with Et0Ac. Dry the organic phase over magnesium
sulfate, filter,
and concentrate under reduced pressure. Purify the residue via silica gel
flash
chromatography using a gradient of 0 to 80% Et0Ac in heptane to give the title
compound. ES-MS m/z 433 (M+H).
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Preparation 73
Methyl 2-(54-cyano-16-methy1-3,9-dioxa-1,2(1,3),5(1,2)-
tribenzenacyclononaphane-14-
ypacetate
NC
0
CO2Me
0
Prepare the title compound essentially as described in Preparation 66 using
methyl
2421312-[(3-bromophenoxy)methyl]-5-cyano-phenyl]propoxy]-5-methyl-4-(4,4,5,5-
tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate, stirring the reaction
mixture at 40 C
for 1 h. Purify the mixture via silica gel flash chromatography eluting with a
gradient of 0
to 20% Et0Ac in DCM to afford the title compound. ES-MS nilz 428 (M+H).
Preparation 74
Methyl 2-(16-methy1-54-(trifluoromethyl)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetate
F3C
0
CO2Me
0 N
Prepare the title compound essentially as described in Preparation 66 using
methyl
2-1213-12-1(6-bromo-2-pyridyl)oxymethy1]-5-(trifluoromethyl)phenyl]propoxy]-5-
methy1-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)phenyl]acetate, stirring
the
reaction at 40 C for 1 h. Purify the title compound via silica gel flash
chromatography
eluting with a gradient of 0 to 20% Et0Ac in DCM. ES-MS nilz 472 (M+H).
Preparation 75
2-(54-Cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetic acid
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NC
0
CO2H
0 N
To a mixture of ethyl 2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetate (290 mg, 0.649 mmol) in
ACN:water (5 mL: 0.5 mL) add 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-
a]pyrimidine
(0.30 g, 2.0 mmol). Stir the reaction mixture at RT for 15 h. Adjust the pH of
the reaction
mixture to pH 7 with 1.0 M aqueous citric acid solution and concentrate under
reduced
pressure to remove volatiles. Dilute the residue with Et0Ac (100 mL) and wash
with
water (50 mL) and saturated aqueous NaCl (50 mL). Dry the organic phase over
sodium
sulfate, filter, and concentrate under reduced pressure to give 238 mg of the
title
compound (88%), which is used in crude form in Preparations 85 and 86. ES-MS
m/z 419
(M+H).
Preparation 76
2-(54-Chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-
14-
yl)a.ceti c acid
CI
, 0
CO2H
0 fµL.
To a mixture of methyl 2-(54-chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetate (152 mg, 0.36 mmol) in ACN (3.6 mL) and
THE'
(3 mL) add an aqueous solution of lithium hydroxide (1.0 M, 1.1 mL, 1.1 mmol).
Stir the
mixture at 40 C for 2 h. Quench the reaction with aqueous citric acid
solution (1.0 M, 2.2
nil-) and then dilute with Et0Ac. Remove the aqueous layer and extract with
Et0Ac (2 x
3 mL). Dry the combined organic phase over magnesium sulfate, filter, and
concentrate
under reduced pressure to give 150 mg of the title compound (100%), which is
used in
crude form in Preparation 87. ES-MS m/z 410 (M+H).
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Preparation 77
2-(54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
NC
, 0
CO2 H
0
Prepare the title compound essentially as described in Preparation 75 starting
with
methyl 2-(54 cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)acetate. After the reaction is complete, cool to RT and neutralize with an
aqueous
solution of citric acid. Filter the resulting precipitate and dry the
resulting filter cake
under vacuum to give the title compound (100%). ES-MS ne/z 401 (M+H)
Preparation 78
2-(54-Cyano-16-fluoro-9-oxa-3-aza-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid
NC
, 0
CO2H
H N N
,
1
To a suspension of ethyl 2-(54-cyano-16-fluoro-9-oxa-3-aza-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetate (122 mg, 027 mmol) in ACN
(3
mL), THE (1 mL) and water (1 mL), add 1,5,7-triazabicyclo[4.4.0]dec-5-ene (116
mg,
0.81 mmol). Stir at 45 C for 2 h, cool to RT and quench with 1M citric acid
solution (2
mL). Extract with Et0Ac (3 >< 3 mL). Combine organics, wash with water and
brine, dry
over magnesium sulfate, filter, and concentrate under reduce pressure, to give
a waxy pale
yellow solid (115 mg, 100%) ES-MS ni/z 418 (M+H).
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Preparation 79
2-(54-Cyano-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetic acid
N C
Th1II
0
CO2 H
0 N
I
Prepare the title compound essentially as described in Preparation 75 using
methyl
2-(54-cyano-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate. ES-MS m/z 415 (M+H).
Preparation 80
2-(54-Cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-
ypacetic acid
CN
0
CO2H
N
Prepare the title compound essentially as described in Preparation 78 using
ethyl
2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-
yl)acetate. ES-MS nilz 419 (M+H).
Preparation 81
2-(54-Cyano-16-fluoro-3,6,9-trioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid
NC 010
CO2H
0
F
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Prepare the title compound essentially as described in Preparation 75 using
ethyl
2-(54-cyano-1b-fluoro-3,6,9-trioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate, stirring the reaction at 45 C for 3 h.
Quench the
reaction with formic acid to pH 7 and dilute with water. Extract three times
with Et0Ac
and then three times with 3:1 chloroform:isopropanol. Dry the organic phase
over
magnesium sulfate, filter, and concentrate under reduced pressure. Purify the
residue via
silica gel flash chromatography using a gradient of 10 to 80% Et0Ac in DCM to
give the
title compound. ES-MS nilz 421 (M+H).
Preparation 82
2-(54-Cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-
14-yl)acetic acid
NC
0
CO2H
0 N
,
Prepare the title compound essentially as described in Preparation 75 using
ethyl
2-(54-cya.no-16-fluoro-3,7-di oxa.-2(2,6)-pyri di na-1(1,3),5(1 ,2)-di ben
zenacycl ooctaphan e-
14-y1), stirring the reaction at 45 C for 1 h. Quench the reaction to pH 6-7
with formic
acid and extract with Et0Ac, followed by 3:1 chloroform:2-propanol. Dry the
organic
phase over magnesium sulfate, filter, and concentrate under reduced pressure
to give the
title compound (95%), which is used in crude form in Preparation 98. ES-MS
ni,/z 405
(M+H).
Preparation 83
2-(54-Cyano-16-methyl-3,9-dioxa-1,2(1,3),5(1,2)-tribenzenacyclononaphane-14-
yl)acetic
acid
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NC
0
CO2H
0
Prepare the title compound essentially as described in Preparation 75 using
methyl
2-(54-cyano-16-methy1-3,9-dioxa-1,2(1,3),5(1,2)-tribenzenacyclononaphane-14-
ypacetate,
stirring the reaction mixture at 45 C for 1 h. Use the title compound in
crude form in
Preparation 99. ES-MS nilz 412 (M-H).
Preparation 84
2-(16-Methy1-54-(trifluoromethyl)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-y1)acetic acid
F3C 0
CO2H
0 N
-
Prepare the title compound essentially as described in Preparation 75 using
methyl
2-(16-m ethy1-54-(tri fluorom ethyl )-3 , 9 -di oxa-2(2,6)-pyridi na-1 (1,3),5
(1, 2)-
dibenzenacyclononaphane-14-yl)acetate, stirring the reaction at 50 C for 1 h.
Use the
title compound in crude form in Preparation 100. ES-MS 111/Z 458 (M+H).
Preparation 85
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3-((oxetan-2-ylmethypamino)benzoate
ço
NC
H 0
0 N 0
, CO2Me
F
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To a mixture of 2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid, (crude, 0.170 g, 0.361 mmol) and
methyl 4-
amino-3-[[(2S)-oxetan-2-yl]m ethyl aminoThenzoate (prepared essentially as
described in
WO 2020/263695; 100 mg, 0.423 mmol) in pyridine (3 mL) add EDC (125 mg, 0.639
mmol). Stir the reaction mixture at RT for 15 h. Quench the reaction with
saturated
aqueous ammonium chloride to pH 6. Dilute the mixture with water (5 mL) and
extract
with Et0Ac (3 x 10 mL). Wash the combined organics with saturated aqueous
sodium
chloride (10 mL). Dry the organic phase over sodium sulfate, filter, and
concentrate under
reduced pressure. Purify the residue via silica gel chromatography eluting
with a gradient
of 0 to 50% Et0Ac in petroleum ether to give 225 mg of the title compound
(71%). ES-
MS m/z 637 (M+H).
Preparation 86
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-fluoro-5-((oxetan-2-
ylmethyl)amino)benzoate
NC
0 H
0 N 0 10
F CO2 M e
F
To a mixture of 2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid (238 mg, 0.568 mmol) and methyl 4-
amino-
3-fluoro-5-[[(2S)-oxetan-2-ylmethyl]aminoThenzoate (0.20 g, 0.79 mmol) in DMF
(5 mL)
add DIPEA (0.15 mL, 0.86 mmol). Add HATU (0.20 g, 0.53 mmol). Stir at RT for 4
h.
Dilute the reaction with Et0Ac (100 mL) and wash with water (50 mL) and
saturated
aqueous NaC1 (50 mL). Dry the organic phase over sodium sulfate, filter, and
concentrate
under reduced pressure to give 0.46 g of the title compound (99%), which is
used in crude
form in Preparation 102. ES-MS m/z 655 (M+H).
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Preparation 87
Methyl (S)-4-(2-(54-chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononap hane-14-yl)acetami do)-3-((oxetan-2-ylm ethyl )ami
no)benzoate
CO
CI H
0N2> 0 'SI
, CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic
acid and methyl 4-amino-341(2S)-oxetan-2-yl]methylaminoThenzoate (prepared
essentially as described in WO 2020/263695), stirring the reaction for 16 h
before
workup. The title compound is used in crude form in Preparation 104. ES-MS
111/Z 628
(M+H).
Preparation 88
Methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3-((oxetan-2-ylmethypamino)benzoate
CO
NC
H
0
0I5J>0 410
CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
and methyl 4-amino-3-[[(2S)-oxetan-2-yl]methylaminoThenzoate (prepared
essentially as
described in WO 2020/263695). Obtain the crude title compound and use without
purification in Preparation 105. ES-MS tn/z 619 (M+H)
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Preparation 89
Methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate
CO
N C 0 H N
,
0 N 0 el
0 CO2 M e
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
and methyl 4-amino-3-methoxy-5-[[(2S)-oxetan-2-yl]methylaminoThenzoate
(Preparation
17). Purify the residue via silica gel chromatography using a gradient of 5 to
80% Et0Ac
in DCM to give the title compound. ES-MS m/z 649 (M+H).
Preparation 90
Methyl (S)-5-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-6-((oxetan-2-ylmethypamino)picolinate
CO
N C
0 H
H N
0 N 0
CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
(0.3g, 0.75 mmol) and methyl (S)-5-amino-6-((oxetan-2-
ylmethyl)amino)picolinate. The
title compound is used in the next step (Preparation 107) without
purification. ES-
MS nilz 620 (M+H)
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Preparation 91
Methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-fluoro-5-((oxetan-2-
ylmethyl)amino)benzoate
CO
NC 0
H N
0 N 0 10
CO2 M e
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
and methyl 4-amino-3-fluoro-5-[[(2S)-oxetan-2-ylmethyl]aminoThenzoate. The
title
compound is used in the next step (Preparation 108) without purification. ES-
MS nilz 637 (M+H)
Preparation 92
Methyl (S)-4-(2-(54-cyano-16-fluoro-9-oxa-3-aza-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3 -((oxetan-2-ylmethyl)amino)benzoate
CO
N C 0 NH
HN.IL1J0
, C 02 M e
To a solution of 2-(54-cyano-16-fluoro-9-oxa-3-aza-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid (115 mg, 0.27 mmol) in DIV1F (3.0
mL), add
methyl 4-amino-3-[[(2S)-oxetan-2-yl]methylaminoThenzoate (prepared essentially
as
described in WO 2020/263695) (75 mg, 0.32 mmol), HATU (160 mg, 0.42 mmol) and
DIPEA (0.15 mL, 0.86 mmol). Stir at RT for 2 h, dilute with water (10 mL) and
extract
with Et0Ac (4 x 5 mL). Combine organics, wash with water and brine, dry over
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magnesium sulfate, filter, and concentrate under vacuum to give the title
compound as a
brown solid (160 mg, 79%) ES-MS rn/z 636 (M+H).
Preparation 93
Methyl (S)-4-(2-(54-cyano-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate
ço
1
0 N 0 10
0 CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)acetic acid and methyl 4-amino-3-methoxy-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate (Preparation 17). Use the title compound as a crude
product in
the next step (Preparation 103) without purification. ES-MS m/z 663 (M+H).
Preparation 94
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-ypacetamido)-3-((oxetan-2-ylmethypamino)benzoate
CN
0 N H
0 N 0 10
\ CO2 Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-16-fluoro-3 , 9-dioxa-2(2, 6)-pyridina-1,5 (1,3 )-dib enzenacy
clononaphane-14-
yl)aceti c acid and methyl 4-amino-3-1-1(2S)-oxetan-2-yllmethylaminoThenzoate
(prepared
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essentially as described in WO 2020/263695). The title compound is used in its
crude
form without purification in Preparation 110. ES-MS m/z 637 (M+H).
Preparation 95
Methyl 4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-4(1-ethy1-1H-imidazol-5-
y1)methyl)amino)benzoate
NC
0 H N
0 N 0 10
CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
(100 mg, 0.25 mmol) and methyl 4-amino-3-(((1-ethy1-1H-imidazol-5-
y1)methyl)amino)benzoate (Preparation 19). Stir the reaction at RT for 18 h,
then dilute
with water and Et0Ac, then extract the aqueous layer four times with Et0Ac.
Dry the
organic phase over magnesium sulfate, filter, and concentrate under reduced
pressure to
give the title compound which is used in crude form in Preparation 111. ES-MS
nilz 657
(M+H).
Preparation 96
(S)-2-(54-Cyano-3 ,9-dioxa-2(2,6)-pyridina-1(1,3),5 (1,2)-
dibenzenacyclononaphane-14-
y1)-N-(2-methoxy-6-((oxetan-2-ylmethyl)amino)-4-(1H-tetrazol-5-
y1)phenyl)acetamide
0
H N
0 N 0 N
, 0 sN
H N¨Nr
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
and (S)-3-methoxy-N1-(oxetan-2-ylmethyl)-5-(1H-tetrazol-5-yl)benzene-1,2-
diamine.
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Stir at RT for 67 h. Dilute the mixture with water and Et0Ac and extract the
aqueous
layer with Et0Ac. Dry the organic phase over magnesium sulfate, filter, and
concentrate
under reduced pressure. Purify the residue via silica gel flash chromatography
using a
gradient of 0 to 100% Et0Ac in heptane followed by 0 to 10% Me0H in DCM. ES-MS
nilz 659 (M+H).
Preparation 97
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,6,9-trioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-((oxetan-2-ylmethyl)amino)benzoate
ço
NC 0
H
0 N 0 410
CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-16-fluoro-3 ,6, 9-trioxa-2(2,6)-pyridina-1(1,3),5 (1,2)-dib
enzenacyclononaphane-14-
yl)acetic acid and methyl 4-amino-3-[[(2S)-oxetan-2-yl]methylaminoThenzoate
(prepared
essentially as described in WO 2020/263695), stirring the reaction at RT for
24 h. Dilute
the crude reaction with water and extract three times with Et0Ac. Dry the
organic phase
over magnesium sulfate, filter, and concentrate under reduced pressure to
provide the title
compound, which is used in crude form in Preparation 112. ES-MS in/z 639
(M+H).
Preparation 98
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetamido)-3-((oxetan-2-ylmethyl)amino)benzoate
NC
ei 0
H N
0
0 N 0 el
, CO2 Me
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Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-
yl)aceti c acid and methyl 4-amino-3-[[(2S)-oxetan-2-yl]methylaminoThenzoate
(prepared
essentially as described in WO 2020/263695), stirring the reaction at RT for 2
h. Dilute
reaction with water and Et0Ac and extract the aqueous layer with Et0Ac. Dry
the
organic phase over magnesium sulfate, filter, and concentrate under reduced
pressure to
give the title compound, which is used in crude form in Preparation 113. ES-MS
m/z 623
(M+H).
Preparation 99
Methyl (S)-4-(2-(54-cyano-16-methy1-3,9-dioxa-1,2(1,3),5(1,2)-
tribenzenacyclononaphane-14-yl)acetamido)-3 -methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate
NC
H
0
0 0 1011
0 CO2Me
Prepare the title compound essentially as described in Preparation 85 using
2454-
cy ano-16-methy1-3,9-dioxa-1,2(1,3),5(1,2)-tribenzenacyclononaphane-14-
yl)acetic acid
(Preparation 83) and methyl 4-amino-3-methoxy-5-[[(25)-oxetan-2-
yl]methylaminoThenzoate (Preparation 17). Use the title compound without
purification
in Preparation 114. ES-MS m/z 662 (M+H).
Preparation 100
Methyl (S)-3-methoxy-4-(2-(16-methy1-54-(trifluoromethyl)-3,9-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetamido)-5-((oxetan-2-
ylmethyl)amino)benzoate
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CO
F3C
0 H
0 N 0 I.
0 CO2Me
Prepare the title compound essentially as described in Preparation 85 using 2-
(16-
methy1-54-(trifluoromethyl)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetic acid (Preparation 84) and methyl 4-amino-3-
methoxy-5-[[(2S)-oxetan-2-yl]methylamino]benzoate (Preparation 17). Use the
title
compound as a crude product without purification in Preparation 115. ES-MS
171/1Z 706
(M+H).
Preparation 101
Methyl (S)-2-((54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
NC
0 r 0
0 N N = CO2Me
I F
Stir a solution of methyl (S)-4-(2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetamido)-3-((oxetan-2-
ylmethyl)amino)benzoate (220 mg, 0.295 mmol) in acetic acid (3.0 mL) at 80 C
for 2 h.
Filter the reaction mixture and concentrate under reduced pressure to give 200
mg of the
title compound (87%), which is used in crude form in Example 1. ES-MS nilz 619
(M+H).
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Preparation 102
Methyl (S)-2-((54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacycl on onaphane-14-yl)m ethyl)-4-fluoro-1 -(oxetan-2-y1 m ethyl)-1 H-
benzo[d]imidazole-6-carboxylate
NC
0 r 0
0 N N = CO2Me
I F
Stir methyl (S)-4-(2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-fluoro-5-((oxetan-2-
ylmethyl)amino)benzoate (0.46 g, 0.56 mmol) in acetic acid (5.0 mL) at 55 C
for 15 h.
Concentrate the reaction mixture and dissolve the residue in Et0Ac (100 mL).
Wash the
organic phase with saturated aqueous sodium bicarbonate solution and brine.
Dry the
organic phase over sodium sulfate, filter, and concentrate under reduced
pressure. Purify
the residue via silica gel chromatography using a gradient of 5 to 60% Et0Ac
in hexanes
to give 0.24 g of the title compound (67%). ES-MS m/z 637 (M+H).
Preparation 103
Methyl (S)-2-((54-cyano-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
NC
, 0
r. 0
0 N N 4
CO2Me
,
0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate (Preparation 93) and 1:1 DCM : acetic acid. Purify via
silica
gel flash chromatography using a gradient of 5 to 60% Et0Ac in DCM. ES-MS m/z
645
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(M+H).
Preparation 104
Methyl (S)-2-454-chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
CI
/
0 N N CO2Me
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3-((oxetan-2-ylmethyl)amino)benzoate,
stirring the reaction mixture at 55 C for 3.5 h and then 65 C for 2 h.
Concentrate the
mixture under reduced pressure and azeotrope with ACN. Dissolve the residue in
DCM,
adsorb onto Celite , and purify via silica gel chromatography eluting with a
gradient of 0
to 100% Et0Ac in hexanes. ES-MS nvz 610 (M+H).
Preparation 105
Methyl (S)-2-((54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacycl ononaphane-14-y1 )methyl)-1-(oxetan -2-ylm ethyl )-1H-benzo[d] i
mi dazol e-6-
carboxylate
NC
0
0 N
N CO2Me
,
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3-((oxetan-2-ylmethyl)amino)benzoate
(Preparation 88) in 1:1 dichloroethane : acetic acid. Purify the residue via
silica gel
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chromatography using a gradient of 5 to 60% Et0Ac in DCM to give the title
compound.
ES-MS m/z 601 (M+H).
Preparation 106
Methyl (S)-2-054-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
NC
I
0 N N CO2 Me
0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate in 1:1 dichloroethane : acetic acid. Purify the
residue via silica
gel chromatography using a gradient of 5 to 60% Et0Ac in DCM to give the title
compound. ES-MS m/z 631 (M+H).
Preparation 107
Methyl (S)-2-054-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-3-(oxetan-2-ylmethyl)-3H-imidazo[4,5-
13]pyridine-5-carboxylate
0
0 N N CO2Me
,
I
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-5-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetami do)-6-((oxetan-2-ylmethypami no)pi col i
nate in
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1:1 dichloroethane : acetic acid, increasing the reaction time to 48 h. Purify
the residue
via silica gel chromatography using a gradient of 5 to 60% Et0Ac in DCM to
give the
title compound. ES-MS nilz 602 (M+H).
Preparation 108
Methyl (S)-2-054-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
NC
0
I /
0 N =
CO2Me
I
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3-fluoro-5-((oxetan-2-
ylmethyl)amino)benzoatc (0.38 g, 0.48 mmol) in 1:1 dichlorocthanc and acctic
acid.
Purify the residue via silica gel chromatography using a gradient of 5 to 60%
Et0Ac in
DCM to give 0.24 g of the title compound (67%). ES-MS nilz 619 (M+H).
Preparation 109
Methyl (S)-2-((54-cyano-16-fluoro-9-oxa-3-aza-2(2,6)-pyri di na-1(1,3 ),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
NC
0 C.) . 0
I
HN NI N CO2Me
F
Heat a solution of methyl (S)-4-(2-(54-cyano-16-fluoro-9-oxa-3-aza-2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetamido)-3-((oxetan-2-
ylmethyl)amino)benzoate (160 mg, 0.20 mmol) in 1,2-dichloroethane (1.5 mL) and
acetic
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acid (1.25 mL) at 50 "V for 6 h. Cool reaction mixture to RT, concentrate
solvents under
reduce pressure, and purify the residue via silica gel chromatography using
using a
gradient of 10 to 50% Et0Ac in DCM to provide 70 mg (53%) of the title
compound as a
white solid. ES-MS iniz 618 (M-4-1).
Preparation 110
Methyl (S)-24(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
CN
0
0 N
N
002 Me
Prepare the title compound essentially as described in Preparation 109 using
methyl (S)-4-(2-(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)acetamido)-3-((oxetan-2-ylmethyl)amino)benzoate,
stirring the reaction mixture at 60 C for 5.5 h. Cool the mixture to RT and
concentrate
under reduced pressure. Purify the residue via silica gel chromatography using
using a
gradient of 10 to 30% Et0Ac in DCM. ES-MS nilz 619 (M H).
Preparation 111
Methyl 2-454-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)methyl)-141-ethyl-1H-imidazol -5 -yl)methyl)- 1H-b enzo[d]imidazole-6-
carboxylate
NC
0 N
0 N N 10, C 02 Me
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Prepare the title compound essentially as described in Preparation 101 using
methyl 4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-y1 )acetam i do)-3 -(((1 -ethyl -1H-imi dazol -5-yl)methyl)ami no)benzoate
(Preparation 95),
stirring the reaction at 65 C for 5 h then 80 C for 17 h. Concentrate the
solution and
azeotrope with ACN. Purify the residue via silica gel chromatography using a
gradient of
0 to 100% Et0Ac in heptane then a gradient of 0 to 10% Me0H in DCM. ES-MS m/z
639
(M+H).
Preparation 112
Methyl (S)-2-((54-cyano-16-fluoro-3,6,9-trioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
NC
=
0 N N CO2 Me
F
Prepare the title compound essentially as described in Preparation 101 using
methyl (S)-4-(2-(54-cyan o-16-fluoro-3,6,9-tri oxa-2(2,6)-pyri din a -
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-((oxetan-2-ylmethyl)amino)benzoate,
stirring the reaction at 65 C for 3 h. Concentrate the reaction and azeotrope
with ACN.
Purify the residue via silica gel flash chromatography using a gradient of 0
to 100%
Et0Ac in heptane then a gradient of 0 to 2% Me0H in DCM to give the title
compound.
ES-MS m/z 621 (M+H).
Preparation 113
Methyl (S)-2-((54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[dlimidazole-6-
carboxylate
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NC
0
f". 0
0 N CO2Me
F
Prepare the title compound essentially as described in Preparation 101 using
methyl (S)-4-(2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetamido)-3-((oxetan-2-ylmethypamino)benzoate,
stirring the reaction at 65 C for 1.5 h. Concentrate the solution and
azeotrope with ACN,
then purify the residue via silica gel flash chromatography using a gradient
of 0 to 60%
Et0Ac in heptane to give the title compound. ES-MS nilz 605 (M+H).
Preparation 114
Methyl (S)-2-454-cyano-16-methy1-3,9-dioxa-1,2(1,3),5(1,2)-
tribenzenacyclononaphane-
14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylate
0
I-
NC
0
7
0 N CO2Me
0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-16-methy1-3,9-dioxa-1,2(1,3),5(1,2)-
tribenzenacyclononaphane-14-yl)acetamido)-3 -methoxy -5 -((oxetan-2-
ylm ethyl)amino)b enzoate (Preparation 99) in 1:1 dichloroethane:acetic acid.
ES-
MS nilz 645 (M+H).
Preparation 115
Methyl (S)-4-m eth oxy -2-((16-m ethyl -54-(tri fluorom ethyl )-3,9-di oxa-
2(2,6)-
pyridina- 1(1,3), 5( 1,2)-dibenzenacyclononaphane-1 4-yl)methyl)-1 -(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carb oxylate
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F3C
= C.-7
N CO2 Me
0 N
0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-3 -m ethoxy-4-(2-(16-m ethyl -54-(tri fluoromethyl)-3 ,9-di oxa-
2(2, 6)-pyri di na-
1(1,3), 5(1,2)-dib enzenacycl on on aphan e-14-ypacetam do)-5-((oxetan-2-
ylmethyl)amino)benzoate (Preparation 100) in 1:1 dichloroethane:acetic acid.
Purify the
title compound via silica gel chromatography using a gradient of 80 to 100%
DCM in
hexane. ES-MS 111/7Z 688 (M+H).
Preparation 116
Methyl 2-(4-bromo-2-methylphenyl)acetate
0
0
Add thionyl chloride (2.5 mL, 34.3 mmol) dropwise over 15 min to a 4 C
solution of 2-(4-bromo-2-methylphenyl)acetic acid (5 g, 20.74 mmol) in Me0H
(42 mL).
Stir the reaction for 3 h then evaporate solvents under reduced pressure. To
the residue
add water (50 mL) and saturated aqueous NaHCO3 to bring the solution to pH = 7-
8, then
extract with Et0Ac (3 x 30 mL). Combine the organics, wash with water and
saturated
aqueous NaCl, dry over MgSO4, then filter and concentrate under reduced
pressure to
give the title compound as an orange oil (5.06g, 100%). The title compound
does not
ionize by LCMS and is used in Preparation 117 without further
characterization.
Preparation 117
Methyl- 4-bromo-2-(bromomethyl)phenyl acetate
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Br
02 Me
Transfer a solution of methyl 2-(4-bromo-2-inethylphenyl)acetate (4.03 g, 15.7
mmol) and N-bromosuccinimide (2.66 g, 14.9 mmol) in ACN (85 mL) through a
photochemical flow reactor equipped with 4 x 370 nm lamps and 4 x 440 nm
lamps,
(reactor size = 52mL, residence time = 1.3mL/min, 40 C). Collect the solution
over 2 h,
evaporate the solvent, then add water (20 mL) and MTBE (20 mL) to the residue.
Separate the layers and extract aqueous phase with MTBE (2 x 20 mL). Combine
the
organics, wash with 20% aqueous NaHS03 solution, water, and saturated aqueous
NaC1,
dry over MgSO4, filter and concentrate under reduced pressure. Purify the
residue via
silica gel chromatography using a gradient of 10 to 40% DCM in cyclohexanes to
give
the title compound as a white waxy solid (5.1 g, 85% purity, 75% yield). ES-MS
m/z
338/340/342 (M+NI-14-').
Preparation 118
Methyl 3-fluoro-5-methoxy-4-nitro-benzoate
0
02 N
To a solution of methyl 3,5-difluoro-4-nitro-benzoate (0.3g, 1.38mmo1) in Me0H
(4 mL) add a solution of sodium methylate (25% by mass in Me0H, 0.33 mL, 1.44
mmol), and heat the reaction at 65 C for 2.5 h. Cool the reaction mixture to
RT, then add
water and extract with Et0Ac (3 x 5 mL). Combine the organics, wash with
saturated
aqueous NaCl, dry over MgSO4, then filter and concentrate in vacuo . Purify
the residue
via silica gel chromatography using a gradient of Et0Ac in heptane (0 to 10%)
to give
245 mg (76%) of the title compound as a yellow oil ES-MS m/z 230 (M-41).
Preparation 119
Ethyl 3-fluoro-5-methoxy -4-nitro-benzoate
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F alb CO2Et
02N
o
Add sulfuric acid (2 mL, 3.1 g, 31 mmol) to a solution of 3-fluoro-5-methoxy-4-
nitro-benzoic acid (0.678, 3.1 mmol) in Et0H (10 mL) and heat the mixture to
80 C for
1 h. Quench the reaction mixture with saturated aqueous NaHCO3 and extract
with
DCM. Dry the organic layer over MgSO4, filter, and concentrate to give the
title
compound (0.73 g, 96%) which is used without further purification in
Preparation 121.
ES-MS ni/z 244 (M+H).
Preparation 120
Methyl 3-fluoro-5-(2-methoxyethoxy)-4-nitro-benzoate
0
¨ 9
o o
Suspend sodium hydride (92 mg, 2.30 mmol) in THF (10 ml), then add 2-
methoxyethanol (0.18 mL, 2.31 mmol) and stir at RT for 30 min. Next, add
methyl 3,5-
difluoro-4-nitro-benzoate (0.5g, 2.30 mmol) and stir the mixture at 60 C for
16 h. Dilute
the reaction with water (100 mL) and extract with Et0Ac (3 x 50 mL). Dry the
organics
over Na2SO4, filter, and concentrate. Purify by silica gel chromatography
using a
gradient of 0 to 20% Et0Ac in heptane to give the title compound (225 mg, 40%)
as a
yellow oil. ES-MS nilz 274 (M+H).
Preparation 121
Ethyl 3-methoxy-4-nitro-5-(oxazol-2-ylmethylamino)benzoate
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(-1;
HN 0.2.
02N
Prepare the title compound essentially as described in Preparation 14 using
ethyl
3-fluoro-5-methoxy-4-nitro-benzoate (0.30 g, 1.0 mmol) and 1-(1,3-oxazol-2-
yl)methanamine hydrochloride (0.20 g, 1.0 mmol). Purify the title compound via
silica
gel chromatography using a gradient of 0 to 50% Et0Ac in heptane. ES-MS nilz
322
(M+H).
Preparation 122
Ethyl-4-amino-3-methoxy-5-(oxazol-2-ylmethylamino)benzoate
(1\1;
HN CO2Et
H2N
0
Suspend iron powder (0.33g, 5.8 mmol) and NH4C1 (0.015 g, 0.28 mmol) in water
(4.4 mL) and add acetic acid (0.07 mL, 1.18 mmol). Stir at 50 C for 15 min
then add a
solution of ethyl 3-methoxy-4-nitro-5-(oxazol-2-ylmethylamino)benzoate (0.165
g, 0.514
mmol) in DMF (1.45 mL). Stir the mixture at 50 C for 20 min, cool to RT,
filter the
mixture through Celite and rinse with Et0Ac (100 mL). Wash the organics with
saturated NaHCO3 solution (100 mL) and dry over MgSO4. Filter and concentrate
to
afford the title compound (0.11 g, 74%) as a yellow oil, which is used without
purification in Preparation 169. ES-MS itz/z 292 (M I II).
Preparation 123
Methyl 3-(2-methoxyethoxy)-4-nitro-5-[[(2S)-oxetan-2-y1]-methylamino]benzoate
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H
02N rah,
ry RP- CO2 Me
0
Prepare the title compound essentially as described in Preparation 14 using
methyl
3-fluoro-5-(2-methoxyethoxy)-4-nitro-benzoate and [(2S)-oxetan-2-
yl]methanamine.
Purify the title compound via silica gel chromatography using a gradient of 0
to
30% Et0Ac in heptane. ES-MS in/1z 341 (M+H).
Preparation 124
Methyl 4-amino-3-(2-methoxyethoxy)-5-[[(25)-oxetan-2-yl]methylamino]benzoate
H
H2N 401
0 CO2Me
0
Prepare the title compound essentially as described in Preparation 122 using
methyl 3-(2-methoxyethoxy)-4-nitro-5-(oxetan-2-yl-methylamino)benzoate. The
product
is used without further purification in Preparation 170. ES-MS in/z 311 (M+H).
Preparation 125
4-[(6-Bromo-2-pyridyl)oxymethy1]-3-RE)-2-ethoxyvinylThenzonitrile
NC
0 N Br
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To a solution of 4-[(6-bromo-2-pyridypoxymethy11-3-iodo-benzonitrile (6 g,
14.45 mmol) in 1,4 dioxane (100 mL) under nitrogen bubbling, add Cs2CO3 (9.5
g, 29
mmol), tetrakis(triphenylphosphine)palladium(0) (835 mg, 0.72 mmol) and (E)-1-
ethoxyethene-2-boronic acid pinacol ester (4.2 mL, 18.8 mmol). Heat reaction
mixture
under nitrogen at 90 C for 1 day, then add more
tetrakis(triphenylphosphine)palladium(0) (835 mg, 0.72 mmol) and (E)-1-
ethoxyethene-
2-boronic acid pinacol ester (1 mL, 4.48 mmol). Continue heating the mixture
at 90 C
for 2 days. Cool the mixture, add water (100 mL) and extract with Et0Ac (3 60
mL).
Combine organics, dry over MgSO4, filter, and concentrate under reduce
pressure. Purify
the residue via silica gel chromatography using a gradient of 0 to 10% Et0Ac
in
cyclohexanes to give 3.6 g (60% yield) of the title compound as a pale yellow
solid. ES-
MS nilz. 359/361 (M+H).
Preparation 126
4-[(6-Bromo-2-pyridypoxymethy1]-3-(2-oxoethyl)benzonitrile
NC 0C)
0 N Br
To a solution of 4-1(6-bromo-2-pyridypoxymethy11-3-1(E)-2-
ethoxyvinyl]benzonitrile (3.6 g, 8.5 mmol) in THF (54 mL) add 4M HC1 in 1,4-
dioxane
(21 mL, 85 mmol). Stir the mixture for 20 h at RT. Concentrate the mixture,
add water
(50 mL) and 2M aqueous sodium carbonate until pH = 8, and then extract with
Et0Ac (3
40 mL). Combine the organics, dry over MgSO4., filter, and concentrate under
reduce
pressure to afford the title compound as a pale orange solid (3.9 g, 97%). ES-
MS /viz
331/333 (M+H).
Preparation 127
4-[(6-Bromo-2-pyridyl)oxymethyl]-3-(2-hydroxyethyl)benzonitrile
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NC OH
0 N Br
To a solution of 4-[(6-bromo-2-pyridypoxymethy11-3-(2-oxoethyl)benzonitrile
(3.9 g, 8.24 mmol) in Me0H (60 mL), add sodium borohydride (550 mg, 14.53
mmoL) in
batches. Stir the mixture for 1 h at RT, evaporate solvent, add DCM (30 mL)
and 1M
NaOH solution (10 mL) and stir for 10 min. Separate the phases and extract the
aqueous
phase with more DCM (2 5 mL). Combine the organics, wash with water and
saturated
aqueous NaCl, dry over MgSO4, filter and concentrate under reduced pressure.
Purify the
residue via silica gel chromatography using a gradient of 10 to 40% Et0Ac in
cyclohexanes to give the title compound as a white waxy solid (1.71 g, 60%).
ES-MS nilz
333/335 (M+H).
Preparation 128
2-bromo-4-(hydroxymethyl)benzonitrile
= H
NC alt
Br
Add lithium borohydride in THF (7.7 mL, 15.4 mmol, 2.0 mol/L) to a solution of
ethyl 3-bromo-4-cyanobenzoate (2 g, 7.71 mmol) in anhydrous THF (20 mL) at 0
C
under nitrogen atmosphere. Allow mixture to reach RT and stir overnight.
Remove most
of THE and add citric acid (5% aqueous) carefully at 0 C. Extract the aqueous
layer with
Et0Ac, combine the organic layers, wash with water and saturated aqueous NaC1,
dry
over anhydrous Na2SO4, filter and concentrate under reduced pressure. Purify
the residue
via silica gel chromatography using a gradient of 30 to 100% Et0Ac in
cyclohexanes to
provide the title compound (1.56 g, 92% purity, 88%) as a white solid. 1H NMR
(400
MHz, DMSO) 6 7.91 (d, J= 7.9 Hz, 1H), 7.80 (s, 1H), 7.51 (d, J = 7.5 Hz, 1H),
5.57 (t,
= 5.8 Hz, 1H), 4.59 (d, J = 5.9 Hz, 2H).
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Preparation 129
2-Bromo-4{(6-chloro-2-pyridypoxymethylibenzonitrile
CN
Br
N C
Prepare the title compound essentially as described in Preparation 30 using 2-
5 bromo-4-(hydroxymethyl)benzonitrile and 6-chloropyridin-2-ol Purify the
title
compound via silica gel chromatography using a gradient of 10 to 30% Et0Ac in
cyclohexanes. ES-MS m/z 323, 325, 327 (M+H).
Preparation 130
10 4-[(6-Chloro-2-pyridyl)oxymethyl]-2-(2-hydroxyethyl)benzonitrile
CN
OH
10 N C
Charge a vial with nickel(II) chloride ethylene glycol dimethyl ether complex
(34
mg, 0.15 mmol) and 4,41-di-tert-butyl-2,2'-bipyridine (48 mg, 0.17 mmol).
Purge the vial
with nitrogen and add anhydrous 1,2-dimethoxyethane (3 mL). Stir mixture for
15min.
Charge another vial with Na2CO3 (335 mg, 3.13 mmol), 2-bromo-4-[(6-chloro-2-
pyridyl)oxymethyl]benzonitrile (502 mg, 1.55 mmol) and
(Ir[dF(CF3)ppy]2(dtbpy))PFo
(18 mg, 0.016 mmol). Purge the vial with nitrogen and add anhydrous 1,2-
dimethoxyethane (12 mL), 2-bromoethanol (1.1 mL, 15 mmol),
tris(trimethylsilyl)silane
(740 uL, 2.33 mmol) and the previously prepared Ni catalyst. Bubble the
mixture with
nitrogen for 5 min and irradiate it in EvoluChemTm photoredox box with Kessil
LED light
456 nm with fan overnight. Filter off the solid, wash it with DCM and
concentrate the
filtrate. Purify the residue via silica gel chromatography using a gradient of
0 to 10% of
Et0Ac in DCM as eluent system to provide the title compound (190 mg, 42%) as a
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yellow waxy solid. ES-MS in/z 289, 291 (M+H).
Preparation 131
2-Bromo-6-[(4-fluoro-2-iodo-phenyl)meth oxy]pyridine
11011
B r N 0
To a mixture of (4-fluoro-2-iodo-phenyl)methanol (2.0 g, 7.9 mmol), 2-bromo-6-
fluoro-pyridine (1.4 g, 7.9 mmol), and 1,4-dioxane (25 mL) add potassium tert-
butoxide
(1.20 g, 10.0 mmol). Stir the reaction mixture at 60 C for 16 h. Dilute the
reaction with
Et0Ac (100 mL) and filter through Celite . Wash the filtrate with water (2 50
mL) and
saturated aqueous sodium chloride (50 mL). Dry the organic phase over Na2SO4,
filter,
and concentrate under reduced pressure. Purify the residue via silica gel
flash
chromatography eluting with a gradient of 5 to 50% DCM in hexanes to give 2.16
g of the
title compound (67%). ES-MS in/z 408 and 410 (M+H).
Preparation 132
3 42-[(6-Bromo-2-pyridyl)oxymethyl]-5-fluoro-phenyl]propoxy-tert-butyl-
dimethyl-
silane
0 k
-s,
/\
N
Br
Prepare the title compound essentially as described in Preparation 28 using 2-
bromo-6-1(4-fluoro-2-iodo-phenyl)methoxybyridine. Purify the title compound
via silica
gel flash chromatography eluting with a gradient of 0 to 10% Et0Ac in hexanes
to give
an oil that does not ionize by ES-MS, which is used directly in Preparation
133 without
further identification.
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Preparation 133
342-[(6-Bromo-2-pyridyl)oxymethyl]-5-fluoro-phenyl]propan-1-01
0 H
I
Br
Prepare the title compound essentially as described in Preparation 29 using 3-
12-
[(6-bromo-2-pyri dyl)oxym ethyl ]-5-fluoro-phenyl ]propoxy-tert-butyl -dim
ethyl -silane
(Preparation 132). ES-MS tn/z 340 and 342 (M+H).
Preparation 134
Methyl 243-[tert-butyhdimethyl)silyl]oxypropy1]-6-(trifluoromethyppyridine-3-
carboxylate
C F3
02 Me
Prepare the title compound essentially as described in Preparation 28 using
methyl
2-bromo-6-(trifluoromethyl)pyridine-3-carboxylate. Purify the title compound
via silica
gel chromatography using a gradient of 0 to 10% Et0Ac in DCM ES-MS nilz 378
(M+H).
Preparation 135
[2-[3-[tert-Butyl(dimethyl)silyl]oxypropy1]-6-(trifluoromethyl)-3-
pyridyl]methanol
cF3
/ \
H 0
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Cool a mixture of methyl 243-[tert-butyl(dimethypsilyl]oxypropy11-6-
(trifluoromethyl) pyridine-3-carboxylate (2.0 g, 5.3 mmol) add THF (40 mL) to -
10 C in
an ice/salt bath. To this mixture add lithium aluminum hydride (0.20 g, 5.3
mmol) and
stir with cooling for 1 h. Quench the reaction by dropwise addition of water
(1 mL) then
dilute with Et0Ac (50 mL). Filter the resulting mixture through Celite4' and
rinse with
Et0Ac (100 mL). Wash the filtrate with water (100 mL) and saturated aqueous
NaCl
(100 mL) then dry over Na2SO4. Filter and concentrate to give the title
compound (1.66
g, 84%) as a tan oil, which is used in Preparation 136 without further
purification. ES-
MS nilz 350 (M+H).
Preparation 136
313-1(6-Bromo-2-pyridyl)oxymethy11-6-(trifluoromethyl)-2-pyridyl]propoxy-tert-
butyl-
dimethyl-silane
CF3
N
o
Br
Prepare the title compound essentially as described in Preparation 51 using
[243-
[tert-butyl(dimethyl)silyl]oxypropyl]-6-(trifluoromethyl)-3-pyridylimethanol
and 2-
bromo-6-fluoro-pyridine, stirring the reaction at 60 C for 16 h. Purify the
title
compound via silica gel flash chromatography eluting with a gradient of 5 to
50% DCM
in hexanes. ES-MS nilz 506 and 508 (M+H).
Preparation 137
343-[(6-Bromo-2-pyridyl)oxymethyl]-6-(trifluoromethyl)-2-pyridyl]propan-1-ol
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C F3
0 H
Br
Prepare the title compound essentially as described in Preparation 29 using 3 -
13-
[(6-bromo-2-pyridyl)oxymethy1]-6-(trifluoromethyl)-2-pyridylipropoxy-tert-
butyl-
dimethyl-silane. Purify the title compound via silica gel flash chromatography
eluting
with a gradient of 5 to 50% Et0Ac in hexanes. ES-MS m/z 390 and 392 (M+H).
Preparation 138
(3-Iodo-4-pyridyl)methanol
LJoH
Cool a mixture of methyl 3-iodopyridine-4-carboxylate (5.0 g, 19 mmol) in THE
(40 mL) and Me0H (10 mL) to -10 C using an ice/salt bath, then add sodium
borohydride (1.52 g, 40.2 mmol) and stir with cooling for 1 h. Quench the
reaction by
dropwise addition of water (1 mL) then dilute with Et0Ac (50 mL). Filter the
resulting
mixture through Celite and rinse w/ Et0Ac (100 mL). Wash the filtrate with
water (100
mL) and saturated aqueous NaCl (100 mL) then dry over Na2SO4. Purify the
residue via
silica gel chromatography using a gradient of 5 to 50% (1:4 Me0H : Et0Ac) in
DCM to
give the title compound (1.63 g, 36%) as a tan solid. ES-MS I/1/z 236 (M+H).
Preparation 139
2-Bromo-6-[(3-iodo-4-pyridyl)methoxy]pyridine
Br N 0
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Prepare the title compound essentially as described in Preparation 51 using (3-
iodo-4-pyridyl)methanol and 2-bromo-6-fluoro-pyridine, stirring the reaction
at 60 C for
16 h. Purify the title compound via silica gel chromatography using a gradient
of 5 to
50% Et0Ac in DCM. ES-MS ni/z 390 and 392 (M+H).
Preparation 140
344-[(6-Bromo-2-pyridyl)oxymethyl]-3-pyridyl]propoxy-tert-butyl-dimethyl-
silane
0,
I
Br
Prepare the title compound essentially as described in Preparation 28 using 2-
bromo-6-[(3-iodo-4-pyridyl)methoxy]pyridine. Purify the title compound via
silica gel
chromatography using a gradient of 0 to 80% Et0Ac in hexanes, and use it in
Preparation
141 without further characterization.
Preparation 141
3-[4-[(6-Bromo-2-pyridypoxymethy1]-3-pyridyl]propan-1-ol
OH
'PN Br
Prepare the title compound essentially as described in Preparation 29 using
344-
[(6-bromo-2-pyridyl)oxymethy11-3-pyridyl]propoxy-tert-butyl-dimethyl-silane
(Preparation 140). Purify the title compound via silica gel chromatography
eluting with a
gradient of 5 to 75% (1:4 Me0H : Et0Ac) in DCM. ES-MS in,/z 322 and 324 (M+H).
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Preparation 142
Methyl 244-bromo-24242-[(6-bromo-2-pyridyl)oxymethyl]-5-cyano-
phenyl]ethoxymethyl]phenydacetate
CO2 Me
NC 0 11101
Br
0 N Br
To a solution of 4-[(6-bromo-2-pyridypoxymethy1]-3-(2-
hydroxyethyl)benzonitrile (1 g, 2.85 mmol) and methyl- 4-bromo-2-
(bromomethyl)phenyl acetate (1.65 g, 4.10 mmol) in DCM (15 mL) at 4 C add 2,6-
di-
tert-butylpyridinc (0.93 mL, 4.24 mmol) and silver triflluoromethanesulfonatc
(1.10g,
4.24 mmol). Stir the mixture for 1 h at low temperature, then at RT. After 5
h, add more
silver trifluoromethanesulfonate (220 mg, 0.85 mmol). After 20 h, filter the
reaction
mixture through Celite , rinsing with DCM. Evaporate the filtrate liquids and
purify by
silica gel chromatography using a gradient of 10 to 100% DCM/cyclohexanes to
give the
title compound as a white solid (570 mg, 75% purity, 26% yield). ES-MS nilz
573/575/577 (M+H).
Preparation 143
Methyl 2424242-[(6-bromo-2-pyridyl)oxymethyl]-5-cyano-phenyl]ethoxymethy1]-4-
(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)phenyl]acetate
CO Me
NC 0 lel
0 Br
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To a solution of methyl 244-bromo-24242-[(6-bromo-2-pyridy1)oxymethy11-5-
cyano-phenyl]ethoxymethyl]phenyl]acetate (630 mg, 0.83 mmol, 75% purity) in
anhydrous 1,4-dioxane (8.2 mL) under nitrogen, add bis(pinacolato)diboron (260
mg, 1
mmol), and KOAc (202 mg, 2.01 mmol). After 5 min, add Pd(dppf)C12 DCM complex
(40 mg, 0.048 mmol) and heat reaction mixture at 80 C. After 3 h, cool the
reaction
mixture to RT, then add water (10 mL) and Et0Ac (10 mL). Separate the layers
and
extract the aqueous phase with Et0Ac (2 5 mL). Combine the organics, wash with
water and saturated aqueous NaC1, dry over MgSO4, filter and concentrate under
reduced
pressure to afford the title compound as a brown oil (850 mg, 60% purity),
which is used
in Preparation 150 without further purification. ES-MS m/z 621/623 (M+H).
Preparation 144
Methyl 2-[4-bromo-2-[2-[5-1(6-chloro-2-pyridyl)oxymethy11-2-cyano-
phenyl]ethoxymethyl]phenyllacetate
C 02 M e
CN
Br
10 C
N0
Prepare the title compound essentially as described in Preparation 142 using
446-
chloro-2-pyridyl)oxymethyl]-2-(2-hydroxyethyl)benzonitrile, with the addition
of
activated 3 A molecular sieves to the reaction mixture. Purify the title
compound via
silica gel chromatography using a gradient of 50% to 100% DCM in cyclohexanes.
ES-
MS m/z 529, 531, 533 (M+H).
Preparation 145
Methyl 2-[2-[2-[546-chloro-2-pyridyl)oxymethyl]-2-cyano-phenyllethoxymethy11-4-
(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]acetate
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CO2 Me
CN
0 0
4111 13176
0
CI
Prepare the title compound essentially as described in Preparation 143 using
methyl 244-bromo-24245-[(6-chloro-2-pyridyl)oxymethy1]-2-cyano-
phenyl]ethoxymethyliphenyl]acetate. Upon completion of the reaction, cool to
RI, add
saturated NaHCO3 and Et0Ac and filter the mixture through Celite'. Separate
aqueous
layer and wash organic layer with water and saturated aqueous NaCl, dry over
anhydrous
Na2SO4, filter and remove solvent. Purify the residue via silica gel
chromatography using
a gradient of 0 to 2% Et0Ac in DCM provide the title compound as a colorless
waxy
solid. ES-MS nilz 577 and 579 (M+H).
Preparation 146
Methyl 2-[24342-[(6-bromo-2-pyridyl)oxymethy1]-5-fluoro-phenyl]propoxy]-5-
methyl-
4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate
CO2Me
0
0 Br
Prepare the title compound essentially as described in Preparation 54 using
342-
[(6-bromo-2-pyridyl)oxymethy1]-5-fluoro-phenyl]propan-1-ol and methyl 242-
hydroxy-
5-m ethyl-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborol an-2-yl)phenyl]acetate, and
using a 40%
solution of DEAD in toluene. Purify the title compound via silica gel flash
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chromatography eluting with a gradient of 85 to 100 `)/0 DCM in hexanes. ES-MS
in/z 628
and 630 (M+H).
Preparation 147
Methyl 2-[243-[3-[(6-bromo-2-pyridypoxymethy1]-6-(trifluoromethyl)-2-
pyridyl]propoxy]-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
y1)phenyl]acetate
CO2Me
0 41111
F3 C N
0 N Br
Prepare the title compound essentially as described in Preparation 55 using 3-
13-
[(6-bromo-2-pyridyl)oxymethy1]-6-(trifluoromethyl)-2-pyridylipropan-1-ol and
methyl 2-
[2-hydroxy-5-methy1-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
y1)phenyl]acetate in
1,4-dioxane as solvent. Stir the reaction mixture at RT for 15 h, then quench
the reaction
with Me0H and concentrate under reduced pressure. Purify the title compound
via silica
gel flash chromatography eluting with a gradient of 85 to 100 % DCM in
hexanes. ES-
MS nilz 678 and 680 (M+H).
Preparation 148
Methyl 24243 44-[(6-bromo -2-pyri dypoxymethyl] -3 -pyridyl]propoxy] -5-methyl
-4-
(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenyliacetate
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CO2Me
14c)õ.
N-'""
LL
0 N Br
Prepare the title compound essentially as described in Preparation 55 using
344-
[(6-bromo-2-pyridyl)oxymethy1]-3-pyridyl]propan-1-ol and methyl 2-[2-hydroxy-5-
methy1-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)phenyl]acetate in 1,4-
dioxane as
solvent and stirring the reaction at RT for 15 h. Quench the reaction with
Me0H and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of 85 to 100 % DCM in hexanes to give the title compound. ES-
MS nilz
611 and 613 (M+H).
Preparation 149
Methyl 2-(54-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)-3,9-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetate
CO2 Me
0 N
I
Add bis(pinacolato)diboron (3.93 g, 15.2 mmol) and KOAc (3.04 g, 30.4 mmol)
to a slurry of methyl 2-(51-chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate (4.33 g, 10.1 mmol) in 1,4-dioxane (0.2
L).
Sparge the mixture with nitrogen for 5 min and then add [chloro(2-
dicyclohexylphosphino-2',4',6'-tri-i-propyl -1,1'-biphenyl)(2' -amino-1, 1 '-
biphenyl-2-y1)
palladium(II)] (0.25 g, 0.31 mmol). Stir the mixture at 85 C for 2.5 h under
a positive
pressure of nitrogen, then cool and concentrate under reduced pressure to
remove most of
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the volatiles. Partition the residue between DCM (0.15 L) and water (0.15 L),
separate
the phases and extract the aqueous with further DCM (50 mL). Wash the combined
organic phases with 2 M aqueous K2CO3 (50 mL) then brine (50 mL), then dry
over
MgSO4 and filter. Concentrate the filtrate to 30 mL volume, add Me0H (0.2 L)
and then
concentrate to 60 mL volume. Stir the mixture at ambient temperature for 3 h,
collect the
solid by filtration and wash with Me0H (30 mL). Dry the filter cake under
reduced
pressure at 50 C for 13 h to afford 4.95 g of the title compound (94%) as a
grey solid.
ES-MS in/z 515 and 516 (M+H).
Preparation 150
Methyl 2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-y1) acetate
NC 0
CO2 M e
0 N
I
Prepare the title compound essentially as described in Preparation 68 using
methyl
2424242- [(6-brom o-2-pyri dyl )oxym ethyl] -5-cyan o-phenyl ethoxym ethyl]
tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate (Preparation 143). ES-MS
415
(M+H).
Preparation 151
Ethyl 2-(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-ypacetate
NC
Yi
CO2 Et
0 N
I F
To a round bottom flask under nitrogen atmosphere add 4-[(6-bromo-2-
pyridyl)oxymethy1]-3-(2-hydroxyethyl)benzonitrile (400 mg, 1.20 mmol),
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triphenylphosphine (473 mg 1.80 mmol), and a solution of ethyl 2-[5-fluoro-2-
hydroxy-4-
(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenydacetate (563 mg, 1.44 mmol)
in
anhydrous THF (10 mL). Stir the mixture until solids dissolve and cool down in
an ice
bath. Add to the mixture a solution of di-tert-butyl azodicarboxylate (423 mg,
1.80 mmol)
in THF (1.6 mL). Remove the ice bath and leave the reaction at RT for 2 h. Add
to the
reaction THF (26 mL) and aqueous potassium phosphate (1 M, 7.2 mL) and stir
the
mixture for 5 min. Add Pd(dtbpf)C12 (80 mg, 0.12 mmol) to the reaction, flush
with
nitrogen several times and heat the reaction to 80 C for 3 h. Cool down the
reaction to
RT, dilute with Et0Ac and add Celite.
Stir the mixture for 10 min, filter the mixture through a pad of Celite, and
wash
the Celite pad with Et0Ac. Dry the filtrate over MgSO4, filter, and
concentrate the
filtrate under reduced pressure. Purify the residue via silica gel
chromatography using a
gradient from 0 to 100% of Et0Ac in cyclohexane to give the title product as a
white
solid (150 mg, 28.9%). ES-MS m/z 433 (M+H).
Preparation 152
Methyl 2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-dibenzenacyclononaphane-
14-
yl)acetate
CN
0
CO2Me
0 N
I
Bubble nitrogen for 5 min into a mixture of methyl 2-[2-[2-[5-[(6-chloro-2-
pyri dyl)oxym ethyl ] -2-cy ano-phenyl ethoxym ethyl ] -444,4,5,5 -tetram
ethyl -1,3,2-
dioxaborolan-2-yl)phenyl]acetate (132 mg, 020 mmol, 88 mass%), THF (5 mL) and
aqueous potassium phosphate tribasic (1.0 M, 1 mL, 1.0 mmol). Add XPhos
Pd(crotyl)C1
(Pd-170 catalyst, CAS number 1798782-02-1, 6 mg, 0.009 mmol) and stir the
mixture at
50 C for 50min. Cool the reaction mixture to RT and add water and Et0Ac.
Separate
aqueous layer and wash organic layer with water and saturated aqueous NaCl,
dry over
anhydrous Na2SO4, filter and concentrate. Purify the residue via silica gel
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chromatography using DCM to provide the title compound (51 mg, 61%) as a white
solid.
ES-MS in/z 415 (M+H).
Preparation 153
Methyl 2-(54-fluoro-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
F
CO2Me
0 N
,
Prepare the title compound essentially as described in Preparation 66 using
methyl
2421312-[(6-bromo-2-pyridyl)oxymethy1]-5-fluoro-phenyl]propoxy]-5-methyl-4-
(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenydacetate and stirring the
reaction
mixture under a nitrogen atmosphere at 40 C for 1 h. Purify the title
compound via silica
gel flash chromatography eluting with a gradient of 0 to 20% Et0Ac in DCM. ES-
MS
111/27422 (M+H).
Preparation 154
Methyl 2-(16-methy1-56-(trifluoromethyl)-3,9-dioxa-2(2,6),5(3,2)-dipyridina-
1(1,3)-benzenacyclononaphane-14-y1)acetate
I
..--
CO2Me
I
-,..
Prepare the title compound essentially as described in Preparation 66 using
methyl
2424343-[(6-bromo-2-pyridyl)oxymethy1]-6-(trifluoromethyl)-2-pyridyl]propoxy]-
5-
m ethyl -444,4, 5, 5-tetram ethyl -1,3,2-di oxaborol an -2-y1 )ph enyl ]
acetate, stirring the
reaction at 40 C for 1 h. Purify the title compound via silica gel
chromatography using a
gradient of 0 to 20% Et0Ac in DCM. ES-MS nilz 473 (M+H).
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Preparation 155
Methyl 2-(16-methyl-3,9-dioxa-2(2,6),5(4,3)-dipyridina-1(1,3)-
benzenacyclononaphane-
14-yl)acetate
CO2Me
0 N
Prepare the title compound essentially as described in Preparation 66 using
methyl
2424344-[(6-bromo-2-pyridyl)oxymethy1]-3-pyridyl]propoxy]-5-methyl-4-(4,4,5,5-
tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]acetate and stirring the reaction
at 40 C for 1
h. Purify the title compound via silica gel chromatography using a gradient of
5 to 35%
Et0Ac in DCM. ES-MS rn/z 405 (M-111)
Preparation 156
2-(54-Bromo-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
Br
CO2H
0
Add Me0H (20 mL) and water (5 mL) to a mixture of methyl 245444,4,5,5-
tetramethy1-1,3,2-dioxaborolan-2-y1)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate (445 g, 855 mmol) and cupric bromide
(573 g,
25.7 mmol). Stir the mixture at 80 C for 30 h. Cool the mixture to ambient
temperature,
add 30% aqueous NH4OH and dilute to a final volume of approx. 0.5 L with
water. Stir
the mixture for 0.5 h, collect the solid by filtration and wash with 10%
aqueous NH4OH
(0.1 L) then water (0.1 L). Add THF (0.14 L), Me0H (70 mL) and 1 M aqueous
LiOH
(35 mL) to the damp solid and stir at 60 "V for 3.5 h. Add 1 M aqueous KH2PO4
(0.1 L)
to the mixture and then dilute to a final volume of approx. 1 L with water.
Leave the
mixture to cool naturally with stirring for 1 h, collect the solid by
filtration and wash with
1:4 water:Me0H (0.2 L) and water (0.1 L). Dry the filter cake under reduced
pressure at
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50 C for 16 h to afford 3.66 g of the title compound (92%) as an off-white
solid. ES-MS
rn/z 454 and 456 (M+H).
Preparation 157
2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
y1)
acetic acid
NC 0
CO2 H
0
Prepare the title compound essentially as described in Preparation 78 using
methyl
2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
y1)
acetate. ES-MS in/z 401 (M+H).
Preparation 158
2-(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-
14-yl)acetic acid
NC
CO2H
0 N
,
-õ
Prepare the title compound essentially as described in Preparation 78 using
ethyl
2-(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-
14-yl)acetate. ES-MS nilz 418 (M+H).
Preparation 159
2-(54-Cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-dibenzenacyclononaphane-14-
yl)acetic
acid
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-1 5 0 -
CN
0
CO2 H
0 N
Prepare the title compound essentially as described in Preparation 78 using
methyl
2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-dibenzenacyclononaphane-14-
yl)acetate.
ES-MS m/z 401 (M+H).
Preparation 160
2-(16-Methy1-54-(fluoro)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid
CO2 H
0
Prepare the title compound essentially as described in Preparation 75 using
methyl
2-(16-methy1-54-(fluoro)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate, stirring the reaction at 50 C for 1 h.
The title
compound is used without purification in Preparation 168. ES-MS nilz 408
(M+H).
Preparation 161
Methyl 2-(16-methy1-56-(trifluoromethy1)-3,9-dioxa-2(2,6),5(3,2)-dipyridina-
1(1,3)-
benzenacyclononaphane-14-yl)aceti c acid
F3C
CO2 H
0 N
,
Prepare the title compound essentially as described in Preparation 75 using
methyl
2-(16-methy1-56-(trifluoromethyl)-3,9-dioxa-2(2,6),5(3,2)-dipyridina-1(1,3)-
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benzenacyclononaphane-14-yl)acetate, stirring the reaction at 50 C for 1 h.
The title
compound is used without further purification in Preparation 171. ES-MS m/z
459
(M+H).
Preparation 162
2-(16-Methy1-3,9-dioxa-2(2,6),5(4,3)-dipyridina-1(1,3)-benzenacyclononaphane-
14-
yl)acetic acid
CO2H
0 N
Prepare the title compound essentially as described in Preparation 75 using
methyl
2-(16-methyl -3 ,9-di oxa-2(2,6),5 (4,3)-dipyri di na-1 (1,3)-benzenacycl on
onaphane-14-
ypacetate and stirring the reaction at 50 C for 1 h. Use the title compound
without
further purification in Preparation 172. ES-MS m/z 391 (M+H).
Preparation 163
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate
ço
N C
H N
lei 0
0 NI 0? Mit
CO2 Me
F
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-
yl)acetic acid and methyl 4-amino-3-methoxy-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate stirring the reaction for 16 h. The title compound is
used in the
next step (Preparation 173) without purification. ES-MS nilz 653 (M+H).
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Preparation 164
Methyl (S)-4-(2-(54-bromo-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetami do)-3-((oxetan-2-ylm ethyl )ami
no)benzoate
Br
H N
0 N 0
CO2 Me
Add DMF (33 mL) and pyridine (6 mL) to a mixture of 2-(54-bromo-3,9-dioxa-
2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetic acid (3.10
g, 6.69
mmol) and methyl 4-amino-3-[[(25)-oxetan-2-ylimethylaminoThenzoate (prepared
essentially as described in WO 2020/263695; 1.75 g, 7.41 mmol) and stir for 30
min.
Add 2,4,6-tripropy1-1,3,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxide (1.68
mol/L in
Et0Ac, 10 mL, 16.8 mmol) and stir the mixture for 50 min. Dilute the reaction
mixture
to a final volume of 0.2 L with water and stir for 20 min_ Collect the solid
by filtration
and wash with water (0.1 L). Dry the filter cake under reduced pressure at 50
C for 24 h
to give 4.64 g of the title compound (99%) as a pale pink solid. ES-MS nilz
672 and 674
(M+H).
Preparation 165
Methyl (S)-4-(2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3-((oxetan-2-ylmethypamino)benzoate
ço
NC 0 H
0 N 0 410
CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-y1)
acetic
acid and methyl 4-amino-341(2S)-oxetan-2-yl]methylaminoThenzoate (prepared
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essentially as described in WO 2020/263695). The title compound is used in the
next step
(Preparation 175) without purification. ES-MS nilz 619 (M+H).
Preparation 166
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetamido)-3-((oxetan-2-ylmethyl)amino)benzoate
C:0
NCN H
0 N 0 el
CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-
yl)acetic acid and methyl 4-amino-3-[[(2S)-oxetan-2-yl]methylaminoThenzoate
(prepared
essentially as described in WO 2020/263695). The title compound is used in
crude form
in Preparation 176. ES-MS in/z 623 (M+H).
Preparation 167
Methyl (S)-4-(2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-
14-y1 )acetami do)-3 -((oxetan -2-ylm ethyl )ami no)benzoate
CN 0
0 CO2 Me
H
0 N
0
1
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-dibenzenacyclononaphane-14-yl)acetic
acid and
methyl 4-amino-3-[[(2S)-oxetan-2-yl]methylamino]benzoate (prepared essentially
as
described in WO 2020/263695). The title compound is used in crude form in
Preparation
177. ES-MS nilz 619 (M+H).
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Preparation 168
Methyl (S)-3-methoxy-4-(2-(16-methy1-54-(fluoro)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacycl ononaphane-14-yl)acetami do)-5-((oxetan-2-
ylmethyl)amino)benzoate
ço
0 N 0?
CO2Me
Prepare the title compound essentially as described in Preparation 86 using 2-
(16-
methy1-54-(fluoro)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)acetic acid (Preparation 160) and methyl 4-amino-3-methoxy-5-[[(2S)-oxetan-
2-
yl]methylaminoThenzoate. The title compound is used without purification in
Preparation
178. ES-MS mtz 656 (M+H).
Preparation 169
Ethyl 4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetamido)-3-methoxy-5-((oxazol-2-ylmethyl)amino)benzoate
NC
H
0 N 0 0 411
CO2Et
Prepare the title compound essentially as described in Preparation 85 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
and ethy1-4-amino-3-methoxy-5-(oxazol-2-ylmethylamino)benzoate (Preparation
122).
The title compound is used without purification in Preparation 179. ES-MS m/z
674
(M+H).
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Preparation 170
Methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate
N C
NH
0 N 0 ? 1.1
, CO2 Me
0
Prepare the title compound essentially as described in Preparation 85 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
and methyl 4-amino-3-(2-methoxyethoxy)-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate
(Preparation 124). The title compound is used without further purification in
Preparation
180. ES-MS 111/2 693 (M+H)
Preparation 171
Methyl (S)-3-methoxy-4-(2-(16-methy1-56-(trifluoromethyl)-3,9-dioxa-
2(2,6),5(3,2)-
dipyridina-1(1,3)-benzenacyclononaphane-14-yl)acetamido)-5-((oxetan-2-
ylmethyl)amino)benzoate
ço
F3 CNL
H
0 N 0? 1101
CO2Me
Prepare the title compound essentially as described in Preparation 86 using
methyl
2-(16-methy1-56-(trifluoromethyl)-3,9-dioxa-2(2,6),5(3,2)-dipyridina-1(1,3)-
benzenacyclononaphane-14-y1)acetic acid (Preparation 161) and methyl 4-amino-3
-
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methoxy-5-[[(2S)-oxetan-2-ydmethylaminobenzoate. Use the title compound in
Preparation 183 without further purification_ ES-MS m/z 707 (M+H).
Preparation 172
Methyl (S)-3-methoxy-4-(2-(16-methy1-3,9-dioxa-2(2,6),5(4,3)-dipyridina-1(1,3)-
benzenacyclononaphane-14-yl)acetamido)-5-((oxetan-2-ylmethyl)amino)benzoate
Co
0 N 0?
, CO2Me
Prepare the title compound essentially as described in Preparation 86 using
2416-
methy1-3,9-dioxa-2(2,6),5(4,3)-dipyridina-1(1,3)-benzenacyclononaphane-14-
yl)acetic
acid (Preparation 162) and methyl 4-amino-3-methoxy-5-[[(2S)-oxetan-2-
yl]methylamino]benzoate. Use the title compound in the next step (Preparation
184)
without further purification. ES-MS m/z 639 (M+H).
Preparation 173
Methyl (S)-24(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
NC
40 0
/ 0
1
OJJ N
,
N = CO2Me
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate (from Preparation 163), heating the reaction at 65 C
for 9h.
Cool the mixture to RT and evaporate the solvent under reduced pressure,
adding ACN to
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help remove the acetic acid. Purify the residue via silica gel chromatography
using a
gradient of 0 to 40% Et0Ac in DCM and then 10% Me0H in DCM to give the title
compound as a pale orange solid. ES-MS nv/z 635 (M+H).
Preparation 174
Methyl (S)-24(54-bromo-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
0
0 N N
CO2Me
-
Prepare the title compound essentially as described in Preparation 102 using
methyl (5)-4-(2-(54-bromo-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3-((oxetan-2-ylmethyl)amino)benzoate
and
using 1:1 acetic acid:2-chlorotoluene as solvent. Stir the reaction at 60 C
for 32 h under a
positive pressure of nitrogen. Concentrate the reaction mixture under reduced
pressure.
Dissolve the residue in DCM, concentrate onto diatomaceous earth and purify by
silica
gel chromatography using a gradient of 0-50% Et0Ac in DCM to give the title
compound
as a white solid. ES-MS miz 654 and 656 (M+H).
Preparation 175
Methyl (S)-2-((54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
NC 0
0 N 1=1 CO2 Me
I
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Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-((oxetan-2-ylmethyl)amino)benzoate
in 1:1
1,2-dichloroethane : acetic acid as solvent, heating the reaction at 50 C for
5 h. Cool
reaction mixture to RT, concentrate solvents under reduce pressure, and purify
the residue
via silica gel chromatography using using a gradient of 10 to 50% Et0Ac in DCM
to give
the title compound as a white solid. ES-MS m/z 601 (M+H).
Preparation 176
Methyl (S)-24(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
NC
r 0
0 N CO2Me
,
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetamido)-3-((oxetan-2-ylmethyl)amino)benzoate
in 1:1
1,2-dichloroethane : acetic acid as solvent, heating to 52 C for 4 h. Cool
down the
reaction to RT and remove the solvent under reduced pressure. Purify the
residue via
silica gel chromatography using a gradient from 0 to 100% of Et0Ac in DCM to
give the
title compound as a yellow solid. ES-MS m/z 605 (M+H).
Preparation 177
Methyl (S)-24(54-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-
14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate
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CN
0
0 N
N
CO2 Me
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-
14-yl)acetamido)-3-((oxetan-2-ylmethyl)amino)benzoate in 1:1 1,2-di
chloroethane : acetic
acid as solvent, heating the reaction at 60 C for 5 h. Cool reaction mixture
to RT,
concentrate solvents under reduce pressure, and purify the residue via silica
gel
chromatography using a gradient of 25 to 50% Et0Ac in DCM to give the title
compound
as a white solid. ES-MS ni/z 601 (M+H).
Preparation 178
Methyl (S)-4-methoxy-24(16-methy1-54-(fluoro)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzord]imidazole-6-
carboxylate
r
0 N N 100 002 Me
0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-3-methoxy-4-(2-(16-methy1-54-(fluoro)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetamido)-5-((oxetan-2-
ylmethyl)amino)benzoate in 1:1 dichloroethane : acetic acid. Purify the title
compound
via silica gel chromatography using a gradient of 80 to 100% DCM in Hexane. ES-
MS in/z 638 (M+H).
Preparation 179
Ethyl 2-((54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
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yl)methyl)-4-methoxy-1-(oxazol-2-ylmethyl)-1H-benzo[dlimidazole-6-carboxylate
Nc)NC
ON1J N CO2Et
¨0
Prepare the title compound essentially as described in Preparation 101 using
ethyl
4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-
14-
yl)acetamido)-3-methoxy-5-((oxazol-2-ylmethyl)amino)benzoate in acetic acid.
Concentrate the reaction mixture and precipitate the title compound from
heptane, which
is used in Example 23 without further purification. ES-MS nilz 656 (M+H).
Preparation 180
Methyl (S)-2-054-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate
I /
0 N N = CO2Me
0
0
Prepare the title compound essentially as described in Preparation 101 using
methyl (S)-4-(2-(54-cyano-3,9-di oxa-2(2,6)-pyri dina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate in acetic acid. Purify the title compound via silica
gel
chromatography using a gradient of 0 to 80% Et0Ac in heptane. ES-MS nilz 675
(M+H).
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Preparation 181
Methyl (S)-2-((54-formy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzen acycl ononaphane-14-yl)methyl)-1-(oxetan -2-ylm ethyl )-1H-benzo[d] i
mi dazol e-6-
carboxylate
is" 0
0 N N = CO2 Me
1
Charge a Schlenk tube with bis(acetonitrile)dichloropalladium(II) (11 mg,
0.042
mmol) and butyldi-l-adamantylphosphine (48 mg, 0.13 mmol). Purge the tube with
nitrogen (3 x vacuum/nitrogen cycles), and add 4-methylmorpholine (3 mL, 27.24
mmol).
Purge the tube again with nitrogen while stirring (5 x vacuum/nitrogen
cycles). Close
tube and stir at ambient temperature for lh. Charge a glass pressure vessel
with methyl
(S)-2454-bromo-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-
14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (290 mg,
0.42
mmol), and 4-methylmorpholinc (9 mL, 81.73 mmol, 100 mass%). Cap with a septum
and bubble the mixture with nitrogen while stirring. After 30 min, transfer
the catalyst
suspension to the pressure vessel and purge it three times with synthesis gas
to 80 psi,
then refill to 80 psi with synthesis gas. Stir and heat the mixture overnight
at 105 C. Cool
the reaction mixture to RT and remove the solvent. Partition the residue
between DCM
(20 mL) and 2 M aqueous K2CO3 (20 mL). Separate the organic layer and extract
the
aqueous layer with DCM (10 mL). Combine the organic layers, wash with
saturated
aqueous NaCl (10 mL), filter and concentrate to afford 320 mg orange residue.
Purify the
residue via silica gel chromatography using a gradient of 0 to 50% Et0Ac in
DCM to
give the title compound (250 mg, 89%) as a white solid. ES-MS m/z 604 (M H).
Preparation 182
(S)-2454-Formy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
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0' .0
r 0
0 1=1 N = CO2H
I
Add 1 M aqueous LiOH (1.25 mL, 1.25 mmol) to a stirred suspension of methyl
(S)-2-454-formy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzordlimidazole-6-carboxylate (0.25 g,
0.37
mmol, 90 mass%) in THF (5 mL), and Me0H (2.5 mL). Seal the reaction vessel and
stir
at 60 C for 2 h. Quench reaction with 1 M aqueous K2HPO4 (5 mL), dilute to a
volume
of 60 mL with water and stir the mixture overnight at ambient temperature.
Adjust the
reaction pH to 4 by the addition of 5% aqueous citric acid, dilute with
saturated aqueous
NaC1 (50 mL), and extract with DCM (50 mL) then three times with 1:4
isopropanol :
DCM (50 mL, 25 mL, 25 mL). Combine and concentrate the organic extracts under
reduced pressure at 50 C. Dissolve the residue in 1:1 DCM Me0H, concentrate
onto
diatomaceous earth and then purify by silica gel chromatography using a
gradient of 0 to
20% Me0H in DCM. Concentrate the appropriate fractions under reduced pressure
at 50
C to afford a white residue, then stir the residue in Et0Ac (5 mL) for 0.5 h.
Collect the
solid by filtration and wash with Et0Ac (5 mL). Dry the solid at 45 C under
reduced
pressure for 21 h to afford the title compound (125 mg, 51%) as a white solid.
ES-MS
nilz 590 (M+H).
Preparation 183
Methyl (S)-4-methoxy-2-((16-methy1-56-(trifluoromethyl)-3,9-dioxa-
2(2,6),5(3,2)-
dipyridina-1(1,3)-benzenacyclononaphane-14-y1)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
0 N N CO2 Me
0
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Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-3-methoxy-4-(2-(16-methy1-56-(trifluoromethyl)-3,9-dioxa-
2(2,6),5(3,2)-
dipyri di n a-1(1,3)-benzen acy cl ononaphane-14-yl)acetami do)-5-((oxetan-2-
ylmethyl)amino)benzoate in 1:1 DCM : acetic acid as solvent. Purify the title
compound
via silica gel chromatography using a gradient of 80 to 100% DCM in hexanes_
ES-
MS ililZ 689 (M+H).
Preparation 184
Methyl (S)-4-methoxy-2-416-methy1-3,9-dioxa-2(2,6),5(4,3)-dipyridina-1(1,3)-
benzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-
6-
carboxylate
0 N
N CO2M e
C
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-3-methoxy-4-(2-(16-methy1-3,9-dioxa-2(2,6),5(4,3)-dipyridina-1(1,3)-
benzenacyclononaphane-14-yl)acetamido)-5-((oxetan-2-ylmethyl)amino)benzoate
(Preparation 172) in 1:1 dichloroethane : acetic acid as solvent. Purify the
title compound
via silica gel chromatography using a gradient of 80 to 100% Et0Ac in DCM. ES-
MS m/z 621 (M+H).
Preparation 185
5-(3-Fluoro-4-nitro-pheny1)-1H-tetrazole
H
02N
To a solution of 3-fluoro-4-nitro-benzonitrile (470 mg, 2.8 mmol) and TMSCN
(4.5 mL, 33 mmol) in toluene (9 mL) add tributyltin azide (2 mL, 7 mmol), then
heat in a
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microwave reactor at 150 'V for 2 h. Quench the reaction with saturated
aqueous
NaHCO3 and extract with Et0Ac. Dry the organic phase over MgSO4, filter, and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a mixture of DCM/lVle01T/formic acid (9:1:0.1) to give 586 mg of the
title
compound (99%). ES-MS m/z 210 (M+H).
Preparation 186
24[5-(3-Fluoro-4-nitro-phenyl)tetrazol-1-yl]methoxy]ethyl-trimethyl-silane
NN
N/
02N
To a solution of 5-(3-fluoro-4-nitro-phenyl)-1H-tetrazole (860 mg, 4.1 mmol)
in
THF (12 mL) add sodium hydride (60% in mineral oil, 180 mg, 4.5 mmol) at 0 C.
Add
2-(chloromethoxy)ethyl-trimethyl-silane (0.79 mL, 4.5 mmol) to the mixture and
stir at
RT for 16 h. Quench the reaction with water and extract with Et0Ac. Dry the
organic
phase over MgSO4, filter, and concentrate under reduced pressure. Purify the
residue via
silica gel chromatography eluting with heptane : Et0Ac (8:2) to give 240 mg of
the title
compound (17%). ES-MS miz 377 (M+H).
Preparation 187
Fumaric acid;[(2S)-oxetan-2-yl]methanamine
gLN H2
H 0 0 H
Mix [(2S)-oxetan-2-yl]methanamine (3.6 wt% in Et0H, 1500 g, 620 mmol) and
fumaric acid (72 g, 620 mmol) at 25 C under nitrogen for 36 h. Filter off the
solid and
dry the solid under reduced pressure to give the title compound (65 g, 52%) as
a white
solid. 1H NMR (400.21 MHz, Me0H-d4) 6 6.72 (s, 2H), 5.02 (ddd, J= 14.8, 7.0,
3.7 Hz,
1H), 4.77-4.70 (m, 1H), 4.61 (dt, J= 9.0, 6.1 Hz, 1H), 3.27 (dd, J= 7.1, 13.4
Hz, 1H), 3.16
(dd, 1= 3.7, 13.4 Hz, 1H), 2.87-2.81 (m, 1H), 2.60-2.54 (m, 111).
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Preparation 188
2-Nitro-/V-[[(2R)-oxetan-2-ylimethy1]-5-[1-(2-
trimethylsilylethoxymethyl)tetrazol-5-
yl]aniline
1 ,N
HN N'
02N
¨Si-
Stir a solution of fumaric acid;[(25)-oxetan-2-yl]methanamine (160 mg, 0.79
mmol) and TEA (0.39 mL, 2.8 mmol) in /V,N-dimethylacetamide (2 mL) at RT for 1
h.
Add 2-[[5-(3-fluoro-4-nitro-phenyl)tetrazol-1-yl]methoxy]ethyl-trimethyl-
silane (240 mg,
0.7 mmol) and stir the mixture at 35 C for 16 h. Quench the reaction with
water and
extract with Et0Ac. Dry the organic phase over MgSO4, filter, and concentrate
under
reduced pressure. Purify the residue via silica gel chromatography eluting
with heptane :
Et0Ac (1:1) to give 200 mg of the title compound (70%). ES-MS nilz 429 (M-I-
Na).
Preparation 189
N2-[[(2R)-Ox etan-2 -yl]m ethyl] -4-[1 -(2-trimethyl silyl ethoxym
ethyptetrazol -5-
yl]benzene-1,2-diamine
,N
H N N
\-0
H 2 N
¨Si¨
/
Vigorously stir a mixture of iron (100 mg, 2 mmol), ammonium chloride (7 mg,
0.1 mmol) and acetic acid (30 !IL, 0.5 mmol) in water (3 mL) at 50 C for 15
min. Add 2-
nitro-N-[[(2R)-oxetan-2-ylimethyl] -5 -[1-(2-trimethyl silyl
ethoxymethyl)tetrazol-5-
yl]aniline (100 mg, 0.2 mmol) in DMF (1 mL) and stir the mixture at 50 C for 1
h. Filter
the reaction mixture through Celite pad, then quench with water and extract
with Et0Ac.
Dry the organic phase over MgSO4, filter, and concentrate under reduced
pressure to give
98 mg of the title compound (99%). ES-MS nilz 377 (M+H).
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Preparation 190
(S)-2-(54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
y1)-N-(2-((oxetan-2-ylm ethyl)amino)-4-(1 -((2-(tri methyl silyl)ethoxy)m
ethyl)- 1H-
tetrazol-5-yl)phenyl)acetamide
NC H N
0
N¨N
0 0
I (0
Si
To a solution of 2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid (100 mg, 0.25 mmol), N2-[[(2R)-
oxetan-2-
yl]methy1]-441-(2-trimethylsilylethoxymethyptetrazol-5-yl]benzene-1,2-diamine
(98 mg,
0.26 mmol) and TEA (104 [EL, 0.75 mmol) in DMF (2 mL), add HATU (142 mg, 0.37
mmol). Stir the mixture at RT for 1 h. Quench the reaction with water and
extract with
Et0Ac. Dry the organic phase over MgSO4, filter, and concentrate under reduced
pressure to give 190 mg of the title compound (99%). LC-MS retention time =
2.17 min.
Preparation 191
(S)-14-((1 -(Oxetan-2-ylmethyl)-6-(1-((2-(trimethyl silypethoxy)methyl)-1H-
tetrazol-5 -y1)-
1H-benzo[d]imidazol-2-yl)methyl)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-54-carbonitrile
=-C-10
N¨NNk
N
\ = N'
Si
0 ¨
/
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Prepare the title compound essentially as described in Preparation 102 using
(S)-
2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
y1)-N-
(2-((oxetan-2-ylm ethyl )ami no)-4-(1 -((2-(tri methyl silypethoxy)methyl)-1H-
tetrazol -5-
yl)phenyl)acetamide in 1:1 1,2-dichloroethane : acetic acid as solvent,
heating the
reaction at 85 C for 16 h. Concentrate the mixture under reduced pressure.
Purify the
residue via silica gel chromatography using a gradient of 0 to 100% Et0Ac in
heptane to
give the title compound. ES-MS m/z 741 (M+H).
Preparation 192
1-Bromo-4-(bromomethyl)-2-fluoro-5-iodo-benzene
/101 Br
Br
Add N-bromosuccinimide (26.84 g, 150.8 mmol) to a solution of 4-bromo-5-
fluoro-2-iodotoluene (25 g, 75.4 mmol) in chloroform (30 mL); then, add 2,2'-
azobi s(2-
methylpropionitrile) (1.26 g, 7.54 mmol) and heat the reaction at 80 C for 5
h. Cool to
RT, add saturated solution of NaHCO3 (300 mL) and extract with DCM (200 mL).
Combine the organics, dry over MgSO4, filter, and concentrate under reduced
pressure.
Purify the residue via silica gel chromatography using heptane as eluent to
give 16.04 g
(54% yield) of the title compound. 1H NMR (400.13 MHz, CDC13) 6 8.05 (d, J=
6.8 Hz,
1H), 7.29 (d, J= 9.00, 1H), 4.51 (s, 2H).
Preparation 193
2-(4-Bromo-5-fluoro-2-iodo-phenyl)acetonitrile
0110 CN
To a solution of 1-bromo-4-(bromomethyl)-2-fluoro-5-iodo-benzene (16.04 g,
40.74 mmol) and TMSCN (7.24 mL, 53 mmol) in ACN (110 mL), add slowly TBAF (1
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M in THF, 53 mL, 53 mmol). Heat the reaction at 40 C for 3 h. Evaporate the
solvents
under reduced pressure, dissolve the residue in Et0Ac (150 mL) and wash the
organics
with saturated aqueous NaCl (3 > 50 mL). Dry the organics over MgSO4, filter,
and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of Et0Ac in heptane from 0 to 15% to afford the title
compound as an
orange oil (10.34 g, 69%). ES-MS ni,/z 340/342 (M+H).
Preparation 194
Ethyl 2-(4-bromo-5-fluoro-2-iodo-phenyl)acetate
SI CO2 Et
Br
To a solution of 2-(4-brc-uno-5-fluoro-2-iodo-phenyl)acetonitri1e (10.34 g, 30
43
mmol) in 8 M Et0H in water (92 mL), add sulfuric acid (24 mL) at RT. Heat the
reaction
mixture at 80 "V for 18 h. Cool the mixture to RT, basify the reaction by
addition of
saturated aqueous NaHCO3 up to pH >7, and extract with DCM ( 3 50 mL). Combine
the organics, wash with water and saturated aqueous NaC1, dry over MgSO4,
filter, and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of Et0Ac in heptane from 0 to 10% to give 8.88 g (75%) of the
title
compound as a white solid. ES-MS nilz 387/389 (M+H).
Preparation 195
Ethyl 2-14-bromo-2-12-ethoxyviny1]-5-fluoro-phenyl]acetate
CO2 Et
Br
Add tetrakis(triphenylphosphine)palladium(0) (1.3 g, 1.1 mmol), Cs2CO3 (7.4 g,
23 mmol) and 2-(2-ethoxyviny1)-4,4,5,5-tetramethy1-1,3,2-dioxaborolane (3.3
mL, 15
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mmol) to a solution of ethyl 2-(4-bromo-5-fluoro-2-iodo-phenypacetate (4.37g,
11.31
mmol) in 1,4-dioxane (80 mL) under nitrogen. Heat the mixture at 90 C for 5
h. Dilute
the mixture with water (100 mL) and extract with Et0Ac (100 mL). Combine the
organics, dry over MgSO4, filter, and concentrate under reduced pressure.
Purify the
residue via silica gel chromatography using a gradient of Et0Ac in heptane
from 0 to
10% to give 2.51g (67%) of the title compound as a yellow oil. ES-MS m,/z
331/333
(M+H).
Preparation 196
Ethyl 2[4-bromo-5-fluoro-2-(2-hydroxyethyl)phenyl]acetate
OH
CO2Et
Br
To a solution of ethyl 2-[4-bromo-2-[2-ethoxyviny1]-5-fluoro-phenyl]acetate
(2.51g, 7.59 mmol) in THE (45 mL) at 0 C, add mercuric acetate (6.3 g, 19
mmol) and
stir for 2 h at 0 C. Meanwhile, add sodium borohydride (520 mg, 13.75 mmol)
to a
solution of K2CO3 (60 g) in water (56 mL), and add this mixture to the
previous reaction
with the starting material. Stir the reaction at RT for 40 min, then dilute
with water (50
mL) and extract with Et0Ac (3 x 50 mL). Combine the organics, dry over MgSO4,
filter,
and concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of Et0Ac in heptane from 0 to 25% to afford the title
compound as a
colorless oil (1.31 g, 40% yield). ES-MS m/z 305/307 (M+H).
Preparation 197
4-F ormy1-3-hydroxy-b enzonitrile
NC 0 H
4111
oI
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To a solution of 4-cyano-2-methoxybenzaldehyde (13 g, 79.86 mmol) in DCM
(480 mL), add boron tribromide (100 g, 399.16 mmol) at -10 C in batches. Stir
the
reaction at RT for 3 days, cool to 0 C, and slowly add water (21 mL). Dilute
the reaction
with water (100 mL) and extract with DCM (3 x 100 mL). Combine the organics,
dry
over MgSO4, filter, and concentrate under reduced pressure. Purify the residue
via silica
gel chromatography using a gradient of 0 to 20% Et0Ac in heptane to give 7.61g
(65%)
of the title compound. ES-MS nilz 148 (M+H).
Preparation 198
4-Formy1-3-(2-trimethylsilylethoxymethoxy)benzonitrile
NC OO-
Add DIPEA (9.5 mL, 54 mmol) and 2-(trimethylsilyl)ethoxymethyl chloride (5.3
mL, 30 mmol) to a solution of 4-formy1-3-hydroxy-benzonitrile (4 g, 27.18
mmol) in
DCM (68 mL) and diethyl ether (30 mL). Stir the reaction mixture for 3 h at
RT. Dilute
the reaction with saturated aqueous NH4C1 and extract with DCM (3 x 50 mL).
Combine
the organics, wash with water and saturated aqueous NaCl, dry over MgSO4,
filter, and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of 0 to 20% Et0Ac in heptane to afford the title compound
(5.67 g,
75%). ES-MS nilz 278 (M+H).
Preparation 199
4-(Hydroxymethyl)-3-(2-trimethylsilylethoxymethoxy)benzonitrile
NC
0 H
To a solution of 4-formy1-3-(2-trimethylsilylethoxymethoxy)benzonitrile (5.67
g,
20.4 mmol) in THF (30mL) and Me0H (30 mL) at 0 C, add sodium borohydride (1.6
g,
42 mmol) in batches. Stir the reaction mixture for 1 h, then add water (50 mL)
and extract
with Et0Ac (3 x 50 mL). Combine the organics, wash with water and saturated
aqueous
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NaCl, dry over MgSO4, filter, and concentrate under reduced pressure to give
5.55g
(97%) of the title compound. ES-MS 111/Z 280 (M+H).
Preparation 200
4-[(6-Bromo-2-pyridyl)oxymethy1]-3-(2-trimethylsilylethoxymethoxy)benzonitrile
NC 0 0
0 N Br
Add sodium hydride (60% in mineral oil, 500 mg, 12.5 mmol) to a solution of 4-
(hydroxymethyl)-3-(2-trimethylsilylethoxymethoxy)benzonitrile (2.65 g, 9.48
mmol) in
THF (60 mL) at RT. Stir the mixture for 30 min, then add 2-bromo-6-
fluoropyridine
(1.7g, 9.5 mmol) and heat the reaction at 60 C for 3 h. Cool the reaction to
ambient
temperature, dilute with water (50 mL) and extract with Et0Ac (3 > 50 mL).
Combine the
organics, wash with water and saturated aqueous NaCl, dry over MgSO4, filter,
and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of 0 to 10% Et0Ac in heptane to give 3.18 g (77%) of the
title
compound. 11-1 NMR (400.13Milz, CDC13) 6 7.57 (d, J= 7.6 Hz, 1H), 7.46 (m,
2H), 7.32
(d, ./-= 8 Hz, 1H), 7.11 (d, ,J= 7.2 Hz, 1H), 6.77 (d, J= 8 Hz, 1H), 5.44 (s,
2H), 5.31 (s,
2H); 3.78 (t, .1= 8.4 Hz, 2H), 0.98 (t, .1= 8.4Hz, 2H). 0.02 (s, 9H).
Preparation 201
4-[(6-13romo-2-pyridyl)oxymethy1]-3 -hydroxy-benzonitril e
NC 011 OH
0 N Br
Add carbon tetrabromide (364 mg, 1.1 mmol) to a solution of 4-[(6-bromo-2-
pyridyl)oxymethy11-3-(2-trimethylsilylethoxymethoxy)benzonitrile (3.18 g,
7.31mmol) in
2-propanol (75 mL). Heat the reaction mixture at 80 C for 10 h, then
concentrate
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solvents under reduced pressure and purify the residue via silica gel
chromatography
using a gradient of 0 to 20% Et0Ac in heptane to afford the title compound as
a white
solid (1.72 g, 57%). ES-MS nilz 305/307 (M+1-1).
Preparation 202
Ethyl 244-bromo-24242-[(6-bromo-2-pyridyl)oxymethy1]-5-cyano-phenoxy]ethyl]-5-
fluoro-phenyl]acetate
Br
JCLCO2Et
NC 0
41)
r0 N B
To a solution of ethyl 2-[4-bromo-5-fluoro-2-(2-hydroxyethyl)phenyl]acetate
(519
mg, 1.70 mmol), 4-[(6-bromo-2-pyridyl)oxymethy1]-3-hydroxy-benzonitrile (500
mg,
1.64 mmol) and triphenylphosphine (645 mg, 2.46 mmol) in THE (17 mL), add at 0
C
DEAD (40% in toluene, 0.97 mL, 2.46 mmol) diluted in THF (1 mL). Stir the
reaction
mixture at RT overnight. After 14 h, add more DEAD (40% in toluene, 0.53 mL,
1.36
mmol) diluted in THE (1 mL) at 0 C. After 20 h at RT, dilute the reaction
mixture with
water (25 mL) and extract with Et0Ac (3 x 10 mL). Combine the organics,
filter, and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of 10 to 30% Et0Ac in heptane to give 444 mg (44%) of the
title
compound as a white solid. ES-MS nilz 591/593/595 (M+H).
Preparation 203
Ethyl 2-(54-cyano-16-fluoro-3,6-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-ypacetate
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N C 0
=
CO2 Et
0
F
In two different batches, bubble nitrogen through a solution of ethyl 2-[4-
bromo-
2-[2-[2-[(6-bromo-2-pyri dyl )oxym ethyl ] -5 -cy ano-ph enoxy] ethy1]-5-
fluoro-phenyl ] acetate
(299 mg, 0.50 mmol) in 1,4-dioxane (10 mL) then add hexamethylditin (0.16 mL,
0.76
mmol) and Pd(dppf)C12 DCM complex (100 mg, 0.12 mmol). Heat the reaction
mixture
batches at 100 C for 3h. Cool both batches to RT and combine them. Dilute
with water
and extract three times with Et0Ac. Combine the organics, wash with water and
saturated
aqueous NaCl, dry over MgSO4, filter, and concentrate under reduced pressure.
Purify
the residue via silica gel chromatography using a gradient of 10 to 20% Et0Ac
in heptane
to afford the title compound as a white solid (90 mg, 41%). ES-MS m/z 433
(M+H).
Preparation 204
2-(54-Cyano-16-fluoro-3,6-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-
14-yl)acetic acid
=
N C 0
02 H
0
F
Prepare the title compound essentially as described in Preparation 75 using
ethyl
2-(54-cyano-16-fluoro-3,6-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooetaphane-
14-ypacetate, heating the reaction at 45 'V for 3 h. Add formic acid to the
mixture until
pH = 5-6, dilute with water, and extract three times with 3:1 DCM :
isopropanol.
Combine the organics, wash with water and saturated aqueous NaCl, dry over
MgSO4,
filter, and concentrate under reduce pressure to afford the title compound as
a white solid.
ES-MS m/z 405 (M+H).
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Preparation 205
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,6-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetamido)-3-((oxetan-2-ylmethypamino)benzoate
NC 0N H
0 N 0 411
CO2 Me
1 F
To a solution of 2-(54-cyano-16-fluoro-3,6-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetic acid (82 mg, 0.20 mmol) and methyl 4-
amino-3-
[[(2S)-oxetan-2-yl]methylamino]benzoate (48 mg, 0.20 mmol) in DMIT (2 mL), add
DIPEA ( 0.10 mL, 0.58 mmol) and HATU (115 mg, 0.30 mmol). After 24 h stirring
at
RT, add more DIPEA (0_055 mL, 0,31 mmol) and HATU (60 mg, 0.15 mmol). After 30
h, add water and Et0Ac, and extract the mixture three times with Et0Ac.
Combine the
organics, wash with water and saturated aqueous NaCl, dry over MgSO4, filter,
and
concentrate under reduced pressure to give the title compound (200 mg, >100%)
which is
used without further purification in Preparation 206. ES-MS m/z 623 (M+H).
Preparation 206
Methyl (S)-24(54-cyano-16-fluoro-3,6-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzord]imidazole-6-
carboxylate
NC 0
1011 NI"'
0 NCO2Me
F
Add acetic acid (6 mL) to methyl (S)-4-(2-(54-cyano-16-fluoro-3,6-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclooctaphane-14-ypacetamido)-3-((oxetan-2-
ylmethypamino)benzoate (Preparation 205, 200mg, 0.32 mmol), and stir at 65 C
for 2 h.
Cool the mixture to RT, evaporate the solvent under reduced pressure and
purify the
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residue via silica gel chromatography using a gradient of 0 to 20% EtOAc in
DCM to
give 92 mg (47%) of the title compound. ES-MS nilz 605 (M+H).
Preparation 207
Ethyl 3-amino-5-bromo-pyridine-2-carboxylate
BrnN H2
N CO2Et
Slowly add sulfuric acid (52 mL, 927 mmol) to a solution of 3-amino-5-
bromopyridine-2-carboxylic acid (15 g, 65.66 mmol) in 8M ethanol in water (197
mL).
Heat the reaction at 80 C for 18 h. Cool the mixture to RT, then slowly add
NaOH (2 M
aqueous) until pH = 8, and extract with Et0Ac (3 x 100 mL). Combine the
organics, dry
over MgSO4, filter, and concentrate under reduced pressure to give the title
compound as
a pale-yellow solid (13.93 g, 86%). ES-MS m/z 245/247 (M+H).
Preparation 208
(3 -Amino-5 -bromo -2-pyri dyl)methanol
BrYN H2
OH
To a solution of ethyl 3-amino-5-bromo-pyridine-2-carboxylate (13.93 g, 56.84
mmol) in THF (230 mL) and Me0H (23 mL) at 0 C, add portion wise lithium
borohydride (3.75 g, 172 mmol). Stir the reaction at RT for 1 h. Add saturated
solution of
NaHCO3 and extract with Et0Ac (5 x 100 mL). Combine the organics, dry over
magnesium sulfate, filter, and concentrate under reduced pressure. Add ACN to
the
residue and filter the resulting slurry, washing the solid with ACN, then dry
the solid
under vacuum to afford 9.87 g (81%) of the title compound as a beige solid. ES-
MS miz
203/205 (M+H).
Preparation 209
(5-Bromo-3-iodo-2-pyridyl)methanol
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BrI
OH
Add 4-methylbenzenesulfonic acid, hydrate (27.81 g, 146.2 mmol) to a
suspension
of (3-amino-5-bromo-2-pyridyl)methanol (9.87 g, 48.63 mmol) in ACN (170 mL).
Stir
the mixture for 10 min, then cool to 0 C. Add sodium nitrite (6.72 g, 97.4
mmol) in water
(20 mL) to the mixture, then add potassium iodide (20.48 g, 123.4 mmol) in
water (20
mL). Stir the reaction at 0 C for 10 min, then 2 h at RT. Add a saturated
aqueous solution
of NaHCO3 to the mixture and extract with Et0Ac (3 x 100 mL). Combine the
organics,
wash with 5% aqueous sodium bisulfite, water, and saturated aqueous NaCl, dry
over
MgSO4, then filter and concentrate under reduced pressure. Triturate the
resulting solid
with ACN, then filter and dry the solid under vacuum to give the title
compound as a
brownish solid (10.31 g, 61%). ES-MS m/z 314/316 (M+H).
Preparation 210
[5-Brom o-3-[3-[tert-butyl (di methyl) si 1 yl] oxyprop-1-yny1]-2-pyridyl
]methanol
\Sr<
0' \
Br
,
0 H
Add bis(triphenylphosphine)palladium (II) dichloride (2.64 g, 3.73 mmol),
cuprous iodide (0.71 g, 3.76 mmol) and TEA (31 mL, 225 mmol) to a solution of
(5-
bromo-3-iodo-2-pyridyl)methanol (11.78 g, 37.53 mmol) under nitrogen at RT.
Add tert-
butyildimethyl(2-propynyloxy)silane (10 mL, 48 mmol) then heat the reaction at
40 C
for 20 h. Cool the reaction to RT, add water and brine, then extract the
mixture three
times with Et0Ac. Combine the organics, dry over MgSO4, filter, and
concentrate under
reduced pressure. Purify the residue via silica gel chromatography using a
gradient of 0 to
20% Et0Ac in heptane to give 9.03 g (67%) of the title compound as a brown
oil. ES-MS
m/z 356/358 (M+H).
Preparation 211
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5-[3-[tert-Butyl(dimethypsilylloxyprop-1-ynyl]-6-(hydroxymethyppyridine-3-
carbonitrile
\ NC ,,,
(_, Si ---
-,/ -- \
I
-..
N
0 H
Add zinc cyanide (2.09 g, 17.85 mmol) to a solution of 15-bromo-343-[tert-
butyl(dimethyl)silylioxyprop-1-yny1]-2-pyridyl]methanol (9.03 g, 25.26 mmol)
in DME
(180 mL) under nitrogen, then add tetrakis(triphenylphosphine)palladium(0)
(2.93 g, 2.54
mmol). Heat the reaction at 100 C for 2 h. Cool the mixture to RT, add water
and
saturated aqueous NaCl and extract the mixture three times with Et0Ac. Combine
the
organics, dry over MgSO4, filter, and concentrate under reduced pressure.
Purify the
residue via silica gel chromatography using a gradient of 0 to 25% Et0Ac in
heptane to
afford 4.6 g (60%) of the title compound as a brown oil. ES-MS m/z 303 (M+H).
Preparation 212
6-[(6-Bromo-2-pyridyl)oxymethyl]-543-[tert-butyl(dimethyl)silyl]oxyprop-1-
ynyl ]pyrl di ne-3-carbonitril e
\
NC ,,.1 .--/ -- \
I
-..
N
0 N Br
fj..-
Prepare the title compound essentially as described in Preparation 34 using 5-
[3-
[tert-butyl (dim ethyl )sily1 ]oxyprop-1-ynyl] -6-(hydroxym ethyl )pyri di ne-
3 -carboni trile.
Upon completion, cool the reaction to RT, evaporate the solvent under reduced
pressure
and purify the residue via silica gel chromatography using a gradient of 0 to
15% Et0Ac
in heptane to give the title compound as a pale yellow solid. ES-MS in/z
458/460 (M+H).
Preparation 213
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6-1(6-Bromo-2-pyridyl)oxymethy11-5-(3-hydroxyprop-1-ynyl)pyridine-3-
carbonitrile
0 H
0 N Br
Add 1M TBAF in THE (16.1 mL, 16.1 mmol) to a solution of 6-[(6-bromo-2-
pyridyl)oxymethy1]-543-[tert-butyl(dimethyl)silyl]oxyprop-1-ynyl]pyridine-3-
carbonitrile (6.158, 13.42 mmol) in THF (40 mL). Stir the mixture at RT for 7
h. Add
saturated aqueous solution of NaHCO3 and extract the mixture three times with
Et0Ac.
Combine the organics, wash with water and saturated aqueous NaCl, dry over
MgSO4,
filter, and concentrate under reduced pressure. Purify the residue via silica
gel
chromatography using a gradient of 0 to 50% Et0Ac in heptane to afford 3.04 g
(66%) of
the title compound as a colorless oil ES-MS m/z 344/346 (M+H)
Preparation 214
6-[(6-Bromo-2-pyridypoxymethy1]-5-(3-hydroxypropyl)pyridine-3-carbonitrile
NC
0 H
0 N Br
Add platinum oxide (0.06 g, 0.26 mmol) to a solution of 6-[(6-bromo-2-
pyridyl)oxymethy11-5-(3-hydroxyprop-1-ynyl)pyridine-3-carbonitrile (0.65 g,
1.88 mmol)
in Me0H (18 mL) and two drops of acetic acid. Charge the reaction vessel with
an
atmosphere of hydrogen (15 psi) and stir the mixture at RT for 5 h. Filter the
reaction
through a pad of Celite washing with Me0H and Et0Ac. Evaporate the filtrate
under
vacuum and purify the residue via silica gel chromatography using a gradient
of 0 to 30%
Et0Ac in heptane to give the title compound (0.33 g, 51%). ES-MS nvz 348/350
(M+H).
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Preparation 215
Ethyl 2-[4-bromo-2-[342-[(6-bromo-2-pyridypoxymethy1]-5-cyano-3-
pyridyl]propoxy]-
5-fluoro-phenyl]acetate
0
NC
0
I Br
0
Prepare the title compound essentially as described in Preparation 60 using 6-
[(6-
bromo-2-pyridyl)oxymethy1]-5-(3-hydroxypropyl)pyridine-3-carbonitrile (1.01 g,
2.9
mmol), eliminating the Me0H quench step, giving the title compound as a white
solid.
ES-MS m/z 606/608 (M+H).
Preparation 216
Ethyl 2-(55-cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-dipyridina-1(1,3)-
benzenacyclononaphane-14-yl)acetate
NC
I 0
0
F
To a mixture of ethyl 244-bromo-243-12-[(6-bromo-2-pyridyl)oxymethyl]-5-
cyano-3-pyridyl]propoxy]-5-fluoro-phenyl]acetate (0.75 g, 1.23 mmol), cesium
fluoride
(203 mg, 1.33 mmol), and hexamethylditin (321 mg, 0.98 mmol) in 1,4-dioxane
(55 mL)
under nitrogen, add chloro(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-
bipheny1)[2-
(2'-amino-1,1'-bipheny1)]palladium(II) (XPhos Pd G2, 75.2 mg, 0.094 mmol).
Heat the
reaction mixture at 100 C for 18 h. Cool the mixture to RT, filter through a
pad of
Celiteg, wash with Et0Ac (2 x 100 mL) and Me0H (2 x 100 mL). Evaporate the
filtrate
and purify the residue via silica gel chromatography using a gradient of 0 to
50% Et0Ac
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in heptane to afford 0.27 g (49%) of the title compound as a light-yellow
solid. ES-MS
rn/z 448 (M+H).
Preparation 217
2-(55-Cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-dipyridina-1(1,3)-
benzenacyclononaphane-
14-yl)acetic acid
0
0 H
0 N 0
I F
To a mixture of ethyl 2-(55-cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-dipyridina-
1(1,3)-benzenacyclononaphane-14-yl)acetate (0.12 g, 0.27 mmol) in ACN (3.2
mL), THF
(0.8 mL) and water (0.5 mL), add 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-
a]pyrimidine
(0.075 g, 0.54 mmol). Heat the suspension at 45 C for 2 h. Cool the mixture
to RT, add
formic acid until pH = 4, and extract with Et0Ac (3 x 5 mL). Combine the
organics, wash
with water and saturated aqueous NaCl, dry over MgSO4, filter, and concentrate
under
reduced pressure to give the title compound as a white solid (0.11 g, 98%). ES-
MS in/z
420 (M+H).
Preparation 218
Methyl (S)-4-(2-(55-cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-dipyridina-1(1,3)-
benzenacyclononaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate
ço
NC H
0
N
0 N 0 0
0
F
To a solution of 2-(55-cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-dipyridina-
1(1,3)-
benzenacyclononaphane-14-yl)acetic acid (0.11 g, 0.26 mmol) and methyl 4-amino-
3-
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methoxy-54[(2S)-oxetan-2-yl]methylaminoThenzoate (0.077 g, 0.29 mmol) in DME
(1.3
mL), add DIPEA (0.13 mL, 0.74 mmol) and HATU (0.16 g, 0.40 mmol). Stir the
reaction
at RT for 16 h. Add saturated aqueous NaHCO3 solution and extract with Et0Ac
(2 x 40
mL). Combine the organics, dry over MgSO4, filter, and concentrate under
reduced
pressure to give the title compound as a beige solid (0.22g), which is used in
Preparation
219 without further purification. ES-MS in/z 668 (M+H).
Preparation 219
Methyl (S)-2-((55-cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-dipyridina-1(1,3)-
benzenacyclononaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
NC
I /"*. 0
0
0 N
(-)
F
M e0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(55-cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-dipyri di na-
1(1,3)-
benzenacyclononaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate (Preparation 218) heating the reaction at 100 C for 2
h. Upon
completion, cool the reaction to RT, add ACN and evaporate. Repeat this
operation three
times to ensure acetic acid removal. Purify the residue via silica gel
chromatography
using a gradient of 0 to 100% Et0Ac in heptane to give the title compound as a
light
brown solid. ES-MS m/z 650 (M+H).
Preparation 220
4-[(6-Chloro-2-pyridyl)oxymethyl]-3-iodo-benzonitrile
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N
I I
1401
0
N
CI
Stir a mixture of 4-(bromomethyl)-3-iodo-benzonitrile (20.0 g, 62.1 mmol), 6-
chloropyridin-2-ol (8.45 g, 65.2 mmol) and silver carbonate (17.1 g, 62.0
mmol) in 1,4-
dioxane (400 mL) for 20 h at 70 C. Add more 6-chloropyridin-2-ol (1.61 g,
12.4 mmol)
and silver carbonate (3.5 g, 13 mmol) and stir the mixture for 5 h at 70 C.
Cool the
mixture to RT and filter through a silica gel plug using DCM as eluent to
yield the title
compound as a pale yellow solid (24.6 g, 107%). ES-MS nilz 371.0/373.0 (M+H).
Preparation 221
4[(6-Chloro-2-pyridypoxymethyl]-3-formyl-benzonitrile
I I
0
0
I
CI
To a mixture of 4[(6-chloro-2-pyridyl)oxymethyl]-3-iodo-benzonitrile (15.0 g,
40.5 mmol), 1,4-diazabicyclo[2.2.2]octane (460 mg, 4.06 mmol) and potassium
formate
(6.90 g, 81.2 mmol) in DMF (180 mL) add tert-butyl isocyanide (5.52 mL, 48.6
mmol),
methanesulfonato(tri-t-butylphosphino)(21-methylamino-1,11-bipheny1-2-
yl)palladium(II)
[P(t-Bu)3 Pd G4, 775 mg, 1.29 mmol] and tri-tert-butylphosphonium
tetrafluoroborate
(360 mg, 1.22 mmol). Stir the mixture for 23 h at 75 C. Cool the mixture to
RT and filter
through a silica gel plug using DMF as eluent. Cool the filtrate to 0 C, add
1N HC1 (120
mL) and stir for 15 min at 0 C. Dilute the reaction mixture with water (200
mL) and stir
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for 30 min at RT. Filter the resulting solid to yield the title compound as
pale green solid
(5.4 g, 77% purity, 37% yield). ES-MS tv/z 273.0/275.0 (M+H)
Preparation 222
4-[(6-Chloro-2-pyridyl)oxymethy1]-3-(hydroxymethyl)benzonitrile
I I
OH
0
N
_I I
Cool a solution of 4-[(6-chloro-2-pyridypoxymethy1]-3-formyl-benzonitrile (1 0
g, 3_67 mmol) in Me0H (20 mL) to at 0 C and add sodium borohydride (290 mg,
726
mmol) portionwise. Stir the mixture for 30 min at RT then cool to 0 C. Add to
the
mixture: water (25 mL), then 5% aqueous citric acid solution up to pH = 5,
then more
water (50 mL). Stir the reaction mixture for 30 min at RT. Filter the
resulting solid to
yield the title compound as white solid (901 mg, 89%). ES-MS nilz 275.0/277.0
(M+H).
Preparation 223
3-(Bromomethyl)-4-[(6-chloro-2-pyridypoxymethyl]benzonitrile
I I
=Br
0
N
CI
Cool a solution of 4-[(6-chloro-2-pyridyl)oxymethyl]-3-
(hydroxymethyl)benzonitrile (870 mg, 3.17 mmol) and triphenylphosphine (932
mg, 3.52
mmol) in DCM (20 mL) at 0 C. Add carbon tetrabromide (920 mg, 2.77 mmol) and
stir
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the reaction mixture for 30 min at RT. Filter the reaction mixture through a
silica plug
using DCM as eluent to yield the compound as a pale brown solid (1.25 g; 116%
yield).
ES-MS m/z 337.0/339.0/341.0 (M+H).
Preparation 224
Ethyl 244-bromo-2424[2-[(6-chloro-2-pyridyl)oxymethyl]-5-cyano-
phenyl]methoxy]ethyl]-5-fluoro-phenyl]acetate
0
OEt
0
Br
0
_N
Stir a solution of ethyl 2[4-bromo-5-fluoro-2-(2-hydroxyethyl)phenyl]acetate
(750 mg, 2 46 mmol), 3-(brc-)momethyl)-4-[(6-chloro-2-
pyridyl)oxymethyl]benzonitri1e
(1.24 g, 3.67 mmol) and 2,6-di-tert-butylpyridine (2.3 mL, 9.9 mmol) in DCM
(13.0 mL)
at RT. Add silver trifluoromethanesulfonate (2.60 g, 10.0 mmol) portionwise
and stir the
reaction mixture at RT for 2 h. Filter the mixture and wash the solid with DCM
(50 mL).
Concentrate the filtrate solvents under reduced pressure. Purify the residue
via silica gel
chromatography using a gradient of 0 to 100% Et0Ac in DCM to give the title
compound
as a yellow solid (480 mg, 34%). ES-MS m/z 561/563 (M+H).
Preparation 225
Ethyl 242-124[2-[(6-chloro-2-pyridyl)oxymethy11-5-cyano-phenyllmethoxy]ethy11-
5-
fluoro-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate
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0
0 Et
0
p-o
GI
0
To a mixture of ethyl 244-bromo-2424[2-[(6-chloro-2-pyridyl)oxymethyl]-5-
cyano-phenyl]methoxy]ethyl]-5-fluoro-phenyl]acetate (150 mg, 0.27 mmol),
bis(pinacolato)diboron (60 mg, 0.23 mmol,) and KOAc (54 mg, 0.54 mmol,) in 1,4-
dioxane (2.5 mL) under nitrogen bubbling, add
dichlorobis(tricyclohexylphosphine)palladium(II) (30 mg, 0.04 mmol) and stir
reaction
mixture at 90 C for 30 min. Cool the reaction mixture to RT, concentrate
under reduced
pressure and purify the residue via filtration through a plug of silica gel
using a gradient
of 0 to 100% Et0Ac in DCM as eluent to give the title compound as a pale white
solid
(101 mg, 65%). ES-MS m/z 609/611(M+H).
Preparation 226
Ethyl 2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
NC
lei 0
0
0 N
F
To a mixture of ethyl 2-[2-[2-[[2-[(6-chloro-2-pyridyl)oxymethy1]-5-cyano-
phenyllmethoxy]ethyl]-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
y1)phenyl]acetate (160 mg, 0.26 mmol) in THF (9.0 mL) with nitrogen bubbling
through
the mixture, add 1N potassium phosphate tribasic in water (1.30 mL, 1.30 mmol)
and
chloro(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-bipheny1)[2-(2'-
amino-1, l'-
biphenyl)]palladium(II) (XPhos Pd G2, 21 mg, 0.03 mmol) and stir the reaction
for 30
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min at 70 C. Cool the reaction mixture to RT, then add MTBE (25 mL) and water
(25
mL). Separate the phases and extract the aqueous phase with MTBE (3 20 mL).
Combine organics, wash with water and saturated aqueous NaCl, dry over sodium
sulfate,
filter and concentrate under reduced pressure. Purify the residue by silica
gel
chromatography using a gradient of 0 to 100% Et0Ac in DCM as eluent to give
the title
compound (45 mg, 34%) as a white solid. ES-MS nilz 447.0 (M+H).
Preparation 227
2-(54-Cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetic acid
NC
410 0
0
0 N OH
F
To a mixture of ethyl 2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetate (91 mg, 0.24 mmol) in ACN
(2.7
mL), THF (0.90 mL) and water (0.90 mL) under nitrogen bubbling, add 1,5,7-
triazabicyclo[4.4.0]dec-5-ene (96 mg, 0.67 mmol) portionwise and stir the
reaction for 1 h
at 45 C. Cool the reaction mixture to RT, add water (5 mL), 5% aqueous citric
acid
solution up to pH = 5 and stir the mixture for 15 min at RT. Filter the
resulting solid to
yield the title compound as white solid (72 mg, 85%). ES-MS nilz 419.0 (M+H).
Preparation 228
Methyl (S)-4-(2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-2-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate
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<,s0
N C
H
0 N 0 0
F 0 0
To a solution of 2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid (70 mg, 0.167 mmol) in DMF (2.5 mL),
add
methyl 4-amino-3-methoxy-5-[[(28)-oxetan-2-yl]methylamino]benzoate (50 mg,
0.19
mmol), HATU (96 mg, 0.25 mmol) and DIPEA (0.1 mL, 0.60 mmol,). Stir the
mixture at
RT for 2 h, then add water (10 mL) and Et0Ac (10 mL). Separate the phases and
extract
the aqueous phase with Et0Ac (3 x 10 mL). Combine the organics, wash with 2 M
aqueous Na2CO3, water, and saturated aqueous NaCl, dry over Na2SO4, filter and
concentrate under reduced pressure to give the title compound as a brown solid
(193 mg,
48% purity, 83% yield), which is used without further purification in
Preparation 229.
ES-MS in/z 667.2 (M+H).
Preparation 229
Methyl (S)-2-((54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
NC lei
I / 0
0
0 -
F ¨0
Heat a solution of methyl (S)-4-(2-(54-cyano-16-fluoro-3,7-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetamido)-2-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate (Preparation 228, 190 mg, 0.28 mmol) in 1,2-
dichloroethane
(3.0 mL) and acetic acid (1.5 mL) at 60 C for 6 h. Cool the reaction mixture
to RT,
concentrate the solvents under reduced pressure, and purify the residue via
silica gel
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chromatography using a gradient of 0 to 100% EtOAc in DCM to give the title
compound
(58 mg, 31%) as a white solid_ ES-MS nilz 649.2/650.2 (M+H).
Preparation 230
2-Bromo-4-chloro-6-fluoro-benzaldehyde
CI B r
F 0
To a solution of 1,2-dibromo-5-chloro-3-fluorobenzene (50 g, 170 mmol) in
heptane (130 mL) and THF (210 mL) at -45 C add dropwise isopropylmagnesium
chloride (2 M solution in TI-1F, 94 mL, 188 mmol) keeping the internal
reaction
temperature between -40 C and -45 C. Stir at -40 C for 30 min, then add
dropwise
DMF (66 mL, 853 mmol) and stir at -20 C for 1 h. Warm the reaction mixture up
to 0 C,
add 1 N HC1 until pH = 7 and extract with Et0Ac (3 A 300 mL). Combine the
organics,
wash with water and saturated aqueous NaCl, dry over Na2SO4, filter, and
concentrate
under reduced pressure. Purify the residue via silica gel chromatography using
a gradient
of 0 to 12% Et0Ac in petroleum ether to give the title compound (22.70 g, 52%)
as a
yellow solid. ES-MS m/z 238 (M+H).
Preparation 231
(2-Bromo-4-chloro-6-fluoro-phenyl)methanol
CI Br
F OH
Add sodium borohydride (5.16 g, 134 mmol) to a solution of 2-bromo-4-chloro-6-
fluoro-benzaldehyde (22.70 g, 89.88 mmol) in Me0H (240 mL) at 0 C. Stir at RT
for 2 h.
Cool to 0 C and add 1N HC1 until pH = 7, concentrate most of the solvent and
extract the
mixture with Et0Ac (3 x 150 mL). Combine the organics, wash with water and
saturated
aqueous NaCl, dry over Na2SO4, filter, and concentrate under reduced pressure
to afford
the title compound as an orange solid (22.3 g, 88%). ES-MS m/z 263 (M+Na).
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Preparation 232
1-Brom o-2-(b rom om ethyl)-5-chloro-3 -fluoro-benzene
CI,::
F Br
To a solution of (2-bromo-4-chloro-6-fluoro-phenyl)methanol (22.3 g, 79.2
mmol)
in DCM (220 mL) at 0 C, add dropwise phosphorous tribromide (7.51 mL, 79.2
mmol).
Bring the reaction mixture to RT and stir for 2 h. Concentrate solvents under
reduced
pressure and purify the residue via silica gel chromatography using a gradient
of 0 to 2%
Et0Ac in petroleum ether to give the tile compound (26.83 g, 95%) as a
colorless oil. 1H-
N1V1IR (400 MHz, DMSO-d6) 6 7.75 (t, J= 2 Hz, 1H), 7.63 (dd, J= 9.5, 2 Hz,
1H), 4.70
(d, J= 2 Hz, 2H).
Preparation 233
2-[(2-Bromo-4-chloro-6-fluoro-phenyl)methoxy]-6-chloro-pyridine
CI uoi Br
F 0 CI
To a solution of 1-bromo-2-(bromomethyl)-5-chloro-3-fluoro-benzene (34.1 g,
107 mmol) and 2-chloro-6-hydroxypyridine (57 g, 431 mmol) in ACN (1000 mL),
add
silver carbonate (180 g, 653 mmol). Stir the reaction mixture at 40 C for 36
h. Cool the
mixture to RT, then filter and concentrate the filtrate under reduced
pressure. Purify the
residue via silica gel chromatography using a gradient of 0 to 5% Et0Ac in
petroleum
ether to afford the title compound as a white solid (22.8 g, 52%). ES-MS tn/z
350/352
(M+H).
Preparation 234
2-Chloro-6-[[4-chloro-2-[(E)-2-ethoxyviny1]-6-fluoro-phenyl]methoxy]pyridine
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CI
F 0 N, CI
To a mixture of 242-bromo-4-chloro-6-fluoro-phenypmethoxy]-6-chloro-
pyridine (20.8 g, 50.4 mmol) and cesium carbonate (33 g, 101 mmol) in 1,4-
dioxane (200
mL), add 2-[(E)-2-ethoxyviny1]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (11.8
mL, 55.4
mmol) under nitrogen, then add tetrakis(triphenylphosphine)palladium(0) (6.13
g, 5.0
mmol). Heat the reaction at 90 C for 12 h, then add more 2-[(E)-2-
ethoxyviny1]-4,4,5,5-
tetramethy1-1,3,2-dioxaborolane (5.4 mL, 25 mmol) and
tetrakis(triphenylphosphine)palladium(0) (3.1 g, 2.5 mmol). Stir the mixture
at 90 C for
4 h more. Cool the mixture to RT and concentrate under reduced pressure, then
add water
(150 mL) and extract with Et0Ac (3 x 150 mL). Combine the organics, dry over
Na2SO4,
filter, and evaporate under reduced pressure. Purify the residue via silica
gel
chromatography using a gradient of 0 to 20% DCM in petroleum ether to give the
title
compound (13.71 g, 70%) as a white solid. ES-MS nilz 342 (M+H).
Preparation 235
2-[5-Chloro-2-[(6-chloro-2-pyridyl)oxymethyl]-3-fluoro-phenyflethanol
CI OH
IF ONC
To a solution of 2-chloro-6-[[4-chloro-2-[(E)-2-ethoxyviny1]-6-fluoro-
phenyl]methoxy]pyridine (12.7 g, 35.3 mmol) in THE' (280 mL) and water (280
mL) at 0
C, add mercuric acetate (37.53 g, 117.8 mmol) and stir at 0 C for 3 h. Add
50% aqueous
K2CO3 (190 mL) and sodium borohydride (6 g, 158.59 mmol) to the mixture at 0
C, then
stir at 0 C for 3 h. Add water (200mL) to the mixture and extract with Et0Ac
(3 >< 500
mL). Combine the organics, dry over Na2SO4, filter, and concentrate under
reduced
pressure. Purify the residue via silica gel chromatography using a gradient of
() to 25%
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Et0Ac in petroleum ether to give the title compound as a colorless oil (9.46
g, 78%). ES-
MS m/z 316 (M+H).
Preparation 236
1-Bromo-5-(bromomethyl)-2-fluoro-4-iodobenzene
Br
Br 11.I
Prepare a solution of 5-bromo-4-fluoro-2-iodotoluene (50.2 g, 156 mmol) and N-
bromosuccinimi de (29.2 g, 164 mmol) in ACN (0.8 L). Pump through a
photochemical
reactor consisting of a coiled PFA reaction tube (1/8" o.d., 52 mL volume)
maintained at
40 C and surrounded by an array of four Kessil PR160-370 nm (40 W) and four
Evoluchem 450 nin (30 W) lamps at a flowrale of between 2 mL/min and 3 mL/min.
Once complete, pump ACN (60 mL) through the reactor at the same rate. Stir the
reactor
output and add 20% aqueous sodium bisulfite (0.2 L) and then water to a final
volume of
2 L. Stir the resulting slurry for 30 min at ambient temperature. Collect the
solid by
filtration and wash with water (0.5 L). Dissolve the filter cake in a mixture
of Et0Ac (0.1
L) and heptane (0.4 L) and wash the organic layer with 50 mL portions of
water, saturated
aqueous NaHCO3 and saturated aqueous NaC1, then dry over MgSO4 and filter.
Concentrate the filtrate under reduced pressure at 50 C to afford 35.19 g of
the title
compound (53%, 93% purity) as a cream solid. 1H-NMR (400 MHz, CDC13) 6 7.65
(d, J
= 6.8 Hz, 1H), 7.60 (J= 7.6 Hz, 1H), 4.51 (s, 2H). 19F t1HI-NMIR (386.5 MHz,
CDC13) -
105.55 (s).
Preparation 237
24[242-[(5-Bromo-4-fluoro-2-iodo-phenyl)methoxy]ethyl]-4-chloro-6-fluoro-
phenyl]methoxy]-6-chloro-pyridine
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IF
CI 0
Br
F 0 N CI
Add silver trifluoromethanesulfonate (8.30 g, 32 mmol) to a solution of 2-15-
chloro-2-1(6-chloro-2-pyridyl)oxymethy1]-3-fluoro-phenyllethanol (5.5 g, 16.0
mmol), 1-
bromo-5-(bromomethyl)-2-fluoro-4-iodobenzene (10.85 g, 24.80 mmol) and 2,6-
ditertbuty1-4-methylpyridine (5 g, 24 mmol) in DCM (30 mL). Stir the reaction
at RT for
5 h. Filter the reaction mixture through Celite , dilute with water (100 mL)
and extract
with Et0Ac (3 x 100 mL). Combine the organics, dry over Na2SO4, filter, and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of 0 to 6% Et0Ac in petroleum ether to give the title
compound as a
colorless oil (8.51 g, 76%). ES-MS miz 628/630 (MHH).
Preparation 238
Ethyl 2-[4-bromo-2-[2-[5-chloro-2-[(6-chloro-2-pyridyl)oxymethy1]-3-fluoro-
phenyllethoxymethyl]-5-fluoro-phenyl]acetate
CO2 Et
CI 0
Br
F 0JCI
To a mixture of 241242-[(5-bromo-4-fluoro-2-iodo-phenyl)methoxy]ethyl]-4-
chloro-6-fluoro-phenyl]methoxy]-6-chloro-pyridine (1.49 g, 1.94 mmol) and
chloro[(4,5-
bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2'-amino-1,1'-
biphenyMpalladium(II)
(Xantphos-Pd-G2, 0.2 g, 0.2 mmol) in THE (7 mL) under nitrogen, add (2-ethoxy-
2-
oxoethyl))zinc bromide (0.5 M in THF, 8 mL, 4 mmol). Heat the mixture at 65 C
for 2 h
in a microwave reactor. Cool the mixture to RT, add saturated aqueous NH4C1
solution
(30 mL), then dilute with water (30 mL) and extract with Et0Ac (3 x 50 mL).
Combine
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the organics, dry over Na2SO4, filter, and concentrate under reduced pressure.
Purify the
residue via silica gel chromatography using a gradient of 0 to 15% Et0Ac in
petroleum
ether to afford 0.57 g (45%) of the title compound as a colorless oil. ES-MS
m/z 588/590
(M+H).
Preparation 239
Ethyl 2-(54-chloro-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
CI 0
CO2Et
F 0 N
To a solution of ethyl 2-[4-bromo-2-[2-[5-chloro-2-[(6-chloro-2-
pyridyl)oxymethy1]-3-fluoro-phenyl]ethoxymethyl]-5-fluoro-phenyl]acetate (1.43
g, 2.19
mmol), potassium 2,2-dimethylpropanoate (0.78 g, 5.49 mmol) and
bis(pinacolato)diboron (0.8 g, 3 mmol) in TT-1F (85 mL) under nitrogen, add
chloro(2-
dicyclohexylphosphino-2',4',6'-triisopropy1-1, 1 '-bipheny1)[2-(2'-amino-1, 1'-
biphenyl)]palladium(II) (XPhos Pd G2, 176 mg, 0.21 mmol). Heat the reaction at
55 C
for 4 h. Add potassium phosphate tribasic (1.42 g, 6.57 mmol) in water (6.57
mL) to the
reaction and heat at 55 C for 2 h. Cool the mixture to RT, add water (30 mL),
and extract
with Et0Ac (3 >< 100 mL). Combine the organics, dry over Na2SO4, filter, and
concentrate under reduced pressure. Purify the residue via silica gel
chromatography
using a gradient of 0 to 15% Et0Ac in petroleum ether to give the title
compound as a
beige solid (308 mg, 27%). ES-MS nilz 474 (M+H).
Preparation 240
2-(54-Chloro-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid
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CI 0
CO2H
F 0
Prepare the title compound essentially as described in Preparation 227 using
ethyl
2-(54-chloro-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate. ES-MS m/z 446 (M+1-1).
Preparation 241
Methyl (S)-4-(2-(54-chloro-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate
CI 0
HN
F 0 N 0
CO2Me
F
0
To a solution of 2-(54-chloro-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetic acid (245 mg, 0.54 mmol)
and
HATIJ (356 mg, 0.91 mmol) in DMF (5.4 mL) under nitrogen add methyl 4-amino-3-
(2-
methoxyethoxy)-5-[[(2S)-oxetan-2-yl]methylamino]benzoate (242 mg, 0.70 mmol)
and
DIPEA (0.28 mL, 1.62 mmol). Stir the mixture at RT for 2 h, then add water (10
mL) and
extract with Et0Ac (3 x 25 mL). Combine the organics, wash with water and
saturated
aqueous NaCl, dry over Na2SO4, filter, and concentrate under reduced pressure
to afford
the title compound as a white solid (594 mg, 34% purity), which is used
without further
purification in Preparation 242. ES-MS nilz 738 (M+H).
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Preparation 242
Methyl (S)-24(54-chloro-16,56-ditluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-442-methoxyethoxy)-1-(oxetan-2-ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate
CI 0
ps. 0
F 0 N 441, CO2Me
1 F
0
0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2454-ehloro-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-342-methoxyethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate in 1:1 1,2-dichloroethane : acetic acid as solvent,
heating the
reaction at 60 'V for 6 h. Cool the reaction mixture to RT, evaporate solvents
under
reduced pressure, adding Et0Ac/toluene (1:1) to help removal of acetic acid in
the
concentration. Purify the residue via silica gel chromatography using a
gradient of 0 to
5% Me0II in DCM to give the title compound as a light-yellow solid (64%
purity). ES-
MS nilz 720 (M+H).
Preparation 243
2-Bromo-6-(bromomethyl)nicotinonitrile
LBr
CN
Br
I
Transfer a solution of 2-bromo-6-methylnicotinonitrile (23 g, 113.2 mmol) and
N-
bromosuccinimide (30.8 g, 170 mmol) in ACN (560 mL) through a photochemical
flow
reactor equipped with a 440-460 nM, 200W lamp, (reactor size = 15 m, 15 mL,
flow rate
¨ 1 mL/min, 25 C). Evaporate the reaction solvent and partition the residue
between
water and DCM. Separate the organic layer, wash with saturated aqueous NaCl,
dry over
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anhydrous Na2SO4, filter and remove solvent. Dissolve the residue in THE (400
mL), add
diethyl phosphite (8.63 mL, 65.8 mmol) and DIPEA (17.8 mL, 99.0 mmol) under N2
at 0
C for 0.5 h. Warm the reaction mixture to RT and stir overnight to give a
black solution.
Partition the reaction mixture between water and Et0Ac. Separate the organic
layer,
wash with saturated aqueous NaCl, dry over anhydrous Na2SO4, filter and
concentrate
under reduced pressure. Purify the residue by silica gel chromatography using
a gradient
of 0 to 25% Et0Ac in petroleum ether to provide the title compound (28.45 g,
84%) as a
white solid. ES-MS nilz 275, 277, 279 (M+H).
Preparation 244
2-Bromo-6-[(6-chloro-2-pyridyl)oxymethyl]pyridine-3-carbonitrile
Br CI
Add silver carbonate (10.5 g, 37.3 mmol) to a solution of 2-bromo-6-
(bromomethyl)nicotinonitrile (1.84 g, 6.33 mmol) and 2-chloro-6-
hydroxypyridine (3.35
g, 25.3 mmol) in ACN (150 mL) at RT. Stir the mixture at 60 C for 48 h.
Filter off the
solid and concentrate filtrate under reduced pressure. Purify the residue by
silica gel
chromatography using a gradient of 0 to 23% Et0Ac in petroleum ether to
provide the
title compound (957 mg, 91 wt% pure, 42%) as a white solid. ES-MS iniz 324,
326, 328
(M+H).
Preparation 245
6-[(6-Chloro-2-pyridyl)oxymethy1]-2-[(E)-2-ethoxyvinyl]pyridine-3-carbonitrile
0)
--ON
NC / N
\ 0 CI
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Add 2-1(E)-2-ethoxyviny1]-4,4,5,5-tetramethy1-1,3,2-dioxaborolane (13.5 mL,
63.4 mmol) and 1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
DCM
complex (4.7 g, 5.65 mmol) to a mixture of 2-bromo-6-[(6-chloro-2-
pyridyl)oxymethyl]pyridine-3-carbonitrile (20.4 g, 56.6 mmol, 91 wt% pure) and
potassium phosphate tribasic (24.5 g, 113 mmol) in 1,4-dioxane (200 mL) and
water (60
mL). Purge the mixture with nitrogen and stir at 90 C for 4 h. Cool the
mixture to RT,
dilute with water (250mL) and extract with Et0Ac (250 mL 4). Combine the
organic
layers, dry over anhydrous Na2SO4, filter and remove solvent. Purify the
residue by silica
gel chromatography using a gradient of 0 to 20% Et0Ac in petroleum ether to
provide the
title compound (15.7g, 83 wt% purity, 73%) as a yellow solid. ES-MS m/z 316,
318
(M+H).
Preparation 246
6-[(6-Chloro-2-pyridyl)oxymethyl]-2-(2-hydroxyethyl)pyridine-3-carbonitrile
HO
NC z N
NcI
Prepare the title compound essentially as described in Preparation 235 using 6-
1(6-
chloro-2-pyridyl)oxymethy1]-2-1(E)-2-ethoxyvinyl]pyridine-3-carbonitrile. Upon
completion of the reaction, filter off a solid and wash with Et0Ac. From the
filtrate,
separate the organic layer and extract the aqueous layer with Et0Ac three
times. Combine
the organic layers, dry over anhydrous Na2SO4, then filter and concentrate
under reduced
pressure. Purify the residue by silica gel chromatography using a gradient of
0 to 45%
Et0Ac in petroleum ether to provide title the compound as a light-yellow
solid. ES-MS
nilz 290, 292 (M+H)
Preparation 247
Methyl 244-bromo-24246-[(6-chloro-2-pyridyl)oxymethy1]-3-cyano-2-
pyridyliethoxymethyliphenyl]acetate
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Br
0
0
N
0
Prepare the title compound essentially as described in Preparation 224 using
64(6-
chloro-2-pyridyl)oxymethy1]-2-(2-hydroxyethyl)pyridine-3-carbonitrile and
methyl 244-
bromo-2-(bromomethyl)phenyl]acetate, stirring the reaction at 40 'V overnight.
Upon
completion of the reaction, remove ACN under reduced pressure, dilute the
residue with
water and extract with Et0Ac. Dry the organics over anhydrous Na2SO4, filter
and
concentrate. Purify the residue by preparative HPLC [column: Phenomenex Luna
C18
250>< 50 mm, 10 um; mobile phase: 40 to 85% ACN in aqueous formic acid
(0.225%)] to
provide the title compound as light yellow waxy solid. ES-MS iii/z 530, 532,
534 (M+H).
Preparation 248
Methyl 2-(55-cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-benzenacyclononaphane-
14-
yl)acetate
CN
0
I
CO2Me
0 N
Prepare the title compound essentially as described in Preparation 216 using
methyl 244-bromo-24246-[(6-chloro-2-pyridyl)oxymethyl]-3-cyano-2-
pyridyllethoxymethyllphenyllacetate as starting material and (2-
dicyclohexylphosphino-
2',4',6'-triisopropy1-1, 1 '-biphenyl) [242' -amino-1,1' -
biphenyl)]palladium(II)
methanesulfonate (XPhos Pd G3) as catalyst, heating the reaction to 110 C
overnight.
When the reaction is complete, concentrate the reaction mixture, add water,
and extract
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three times with Et0Ac. Dry the combined organics over Na2SO4, filter, and
concentrate.
Purify the residue by HPLC [column: Welch Xtimate C18 150 x 40 mm, 10 vim;
mobile
phase: 40 to 80% ACN in aqueous formic acid (0.225%)] to provide the title
compound
as a white solid ES-MS iv/z 416 (M+H).
Preparation 249
2-(55-Cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-benzenacyclononaphane-14-
yl)acetic
acid
CN
I
CO2H
0 N
, -..
I
---
Prepare the title compound essentially as described in Preparation 78 using
methyl
2-(55-cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-benzenacyclononaphane-14-
yl)acetate.
When the reaction is complete, remove the organic solvents and water and
aqueous citric
acid (1 M) to bring the pH to 5-6. Filter the resulting solid and wash with
water to provide
the title compound as a white solid. ES-MS m/z 402 (M+H).
Preparation 250
Methyl (S)-4-(2-(55-cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-
benzenacyclononaphane-
14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-ylmethyl)amino)benzoate
CN
..,),.......-...,.....õ.0
I .. HN =c--A
H --TN
N
0 411 ib--'
0 N
CO2Me
I
..--
r)
0
...-
Prepare the title compound essentially as described in Preparation 86 using 2-
(55-
cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-benzenacyclononaphane-14-yl)acetic
acid and
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methyl 4-amino-3-(2-methoxyethoxy)-54[(2S)-oxetan-2-yllmethylamino]benzoate,
stirring the reaction at RT overnight. Dilute the reaction mixture with water
and extract
three times with Et0Ac. Combine organic layers, wash with water and saturated
aqueous
NaCl, dry over Na2SO4, filter, and concentrate under vacuum to give the title
compound
as an orange waxy solid which is 54 wt% pure. ES-MS m/z 694 (M+H).
Preparation 251
Methyl (S)-2-((55-cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-
benzenacyclononaphane-
14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylate
CN
I rsi
0
N
I
0 N
N CO2Me
I 411
...-
_/-0
0
/
Prepare the title compound essentially as described in Preparation 109 using
methyl (S)-4-(2-(55-cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-
benzenacyclononaphane-
14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-ylmethyl)amino)benzoate (54
wt%
pure), stirring under nitrogen atmosphere at 60 C for 6 h. Purify by silica
gel
chromatography using a gradient of 0 to 6% Me0H in DCM providing the title
compound
as an orange waxy solid which is 63 wt% pure. ES-MS ni/z 676 (M+H).
Preparation 252
Methyl 2-chloro-54(6-chloropyridin-2-yl)oxy)methyl)benzoate
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CI 0
0
CI
To a solution of methyl 5-(bromomethyl)-2-chlorobenzoate (25 g, 95 mmol) in
1,4-dioxane (600 mL), add 6-chloropyridin-2-ol (14.2 g, 110 mmol) and silver
carbonate
(53.2g. 193 mmol). Stir the mixture at 60 C for 23 h. Filter the reaction
suspension
through Celite and rinse with Et0Ac. Concentrate the filtrate under reduced
pressure to
give 29.4 g of the title compound (99%), which is used without further
purification in
Preparation 253. ES-MS rrilz 312, 314 (M+H).
Preparation 253
(2-Chloro-5-(((6-chloropyridin-2-yl)oxy)methyl)phenyl)methanol
CI
OOH
0
CI
Cool a solution of methyl 2-chloro-5-(((6-chloropyridin-2-
yl)oxy)methyl)benzoate
(from Preparation 252, 22 g, 71 mmol) in THF (200 mL) to 0 C, then add Red-Al
(60
wt% in toluene, 30 mL, 92 mL) dropwise. Stir the mixture at 0 C for 10 min
then quench
the reaction with Et0Ac (10 mL). Stir the mixture at RT for 2 h, then dilute
the reaction
with water (200 mL) and Et0Ac (200 mL). Extract the aqueous layer with Et0Ac
(2 x
100 mL). Dry the combined organic phases over MgSO4, filter, and concentrate
under
reduced pressure to give 20.7 g of the title compound (100%), which is used
without
further purification in Preparation 254. ES-MS nilz 284, 286 (M+H).
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Preparation 254
2-((3-(Bromomethyl)-4-chlorobenzyl)oxy)-6-chloropyridine
CI
=Br
0
CI
Cool a solution of (2-chloro-5-(((6-chloropyridin-2-
yl)oxy)methyl)phenyl)methanol (from Preparation 253, 1.5 g, 5.3 mmoL) and
triphenylphosphine (2.0 g, 7.5 mmoL) in DCM (35 mL) to 0 'C. Add carbon
tetrabromide
(1.9 g, 5.7 mmoL), stir the reaction mixture at 0 C for 10 min then at RT for
30 min.
Filter the reaction solution through a pad of silica gel and rinse with DCM.
Concentrate
the filtrate under reduced pressure to give 1.8 g of the title compound
(100%), which is
used without further purification in Preparation 255. ES-MS ni/z 345/347/349
(M+1-1).
Preparation 255
Ethyl 2-(4-bromo-2-(2-((2-chloro-5-(((6-chloropyridin-2-
yl)oxy)methyl)benzyl)oxy)ethyl)-5-fluorophenyl)acetate
CO2Et
CI
=0 Br
0
CI
Prepare the title compound essentially as described in Preparation 224 using 2-
((3-
(bromomethyl)-4-chlorobenzyl)oxy)-6-chloropyridine (from Preparation 254) and
ethyl
2[4-bromo-5-fluoro-2-(2-hydroxyethyl)phenyl]acetate, stirring the reaction at
RT for 1 h
15 min. Filter and concentrate the reaction mixture under reduced pressure,
then purify
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via silica gel chromatography using a gradient of 0 to 20% Et0Ac in hexanes to
give the
title compound. ES-MS rn/z 570/572/574 (M+H).
Preparation 256
Ethyl 2-(2-(2-((2-chloro-5-(((6-chloropyridin-2-yl)oxy)methyl)benzypoxy)ethyl)-
5-
fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-y1)phenyl)acetate
CO2Et
CI
xcI
0
0 B176
0
0
To a mixture of ethyl 2-(4-bromo-2-(2-((2-chloro-5-(((6-chloropyridin-2-
yl)oxy)methyl)benzyl)oxy)ethyl)-5-fluorophenyl)acetate (855 mg, 1.5 mmoL),
bis(pinacolato)diboron (480 mg, 1.87 mmoL), KOAc (450 mg, 4.5 mmoL), and
dichlorobis(tricyclohexylphosphine)palladium (II) (225 mg, 0.30 mmoL) add 1,4-
dioxane
(15 mL). Stir the mixture at 90 C for 5 h, then add Pd(dppf)C12 (125 mg, 0.17
mmoL)
and stir the mixture at 90 C for 15 h. Filter the crude mixture through a pad
of silica gel
and rinse with Et0Ac. Concentrate the filtrate under reduced pressure to give
the title
compound, which is used without further purification in Preparation 257. ES-MS
nilz 536
(M+H for boronic acid).
Preparation 257
Ethyl 2-(54-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)acetate
$0
0 0
I F
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To a solution of ethyl 2-(2-(24(2-chloro-5-(((6-chloropyridin-2-
yl)oxy)methyl)benzyl)oxy)ethyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-
y1)phenyl)acetate (from Preparation 256, 925 mg, 1.5 mmoL) and XPhos Pd G2
(145 mg,
0.18 mmoL) in THF (50 mL), add a solution of potassium phosphate (1.6 g, 7.4
mmoL)
in water (5 mL), Stir the reaction mixture at 60 C for 1.5 h. Dilute the
crude reaction
mixture with Et0Ac (50 mL) and 1:1 water : saturated aqueous NaCl (50 mL), and
extract the aqueous layer with Et0Ac (50 mL). Dry the combined organic phases
over
MgSO4, filter, and concentrate under reduced pressure. Purify the residue via
silica gel
chromatography using a gradient of 0 to 40% Et0Ac in hexanes to give 140 mg of
the
title compound (20%). ES-MS m/z 456 (M+H).
Preparation 258
2-(54-Chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-
yl)acetic acid
0
0 15 H
0 N 0
F
Prepare the title compound essentially as described in Preparation 75 using
ethyl
2454-chi oro-16-fluoro-3,7-di oxa-2(2,6)-pyri di n a-1,5(1,3)-di ben zen acycl
on on aph an e-14-
yl)acetate, using 3:3:1 ACN : 1,4-dioxane : water as solvent and heating the
reaction at 50
C for 1 h 20 min. Dilute the reaction with water and quench with 1 M aqueous
citric acid
solution. Collect the precipitated material by filtration and rinse with water
to give the
title compound. ES-MS nilz 428 (M+H).
Preparation 259
Methyl (S)-4-(2-(54-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-ypacetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
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ylmethyl)amino)benzoate
CI
4101 0
H N c 3
0
0 N 41111 0 Si
0 CO2 Me
I F
rj
0
Prepare the title compound essentially as described in Preparation 86 using
2454-
chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-dibenzenacyclononaphane-14-
yl)acetic acid and methyl 4-amino-3-(2-methoxyethoxy)-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate, stirring the reaction at RT for 17 h. Dilute the
reaction with
Et0Ac and water, wash the organic layer with water and back-extract the
aqueous layer
twice with Et0Ac. Dry the combined organic phases over MgSO4, filter, and
concentrate
under reduced pressure to provide the title compound, which is used without
further
purification in Preparation 260. ES-MS m/z 720 (MAT).
Preparation 260
Methyl (S)-2-((54-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate
CI
40 0
0 N Nil .0 CO2Me
I F
0
0
Prepare the title compound essentially as described in Preparation 109 using
methyl (S)-4-(2-(54-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
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ylmethyl)amino)benzoate (from Preparation 259). Purify via silica gel
chromatography
using a gradient of 0 to 100% Et0Ac in hexanes to give the title compound_ ES-
MS nilz
702 (M+H).
Preparation 261
4-[(6-Bromo-5-fluoro-2-pyridyl)oxymethyl]-3-iod o-benzonitrile
NC
10111
0 N Br
I
Prepare two batches on equal scale as follows: add Na2CO3 to a solution of 4-
(bromomethyl)-3-iodo-benzonitrile (22.4 g, 62.6 mmol) in acetone (340 mL) and
water
(340 mL). Stir both batches at 80 C overnight, then combine the two batches.
Concentrate the mixture to remove the acetone, then filter off the solid and
wash with
water. Dry the solid under vacuum, then stir it for 30 min in DCM (70 mL).
Filter the
solid, rinse with DCM, and dry under vacuum to give 4-(hydroxymethyl)-3-iodo-
benzonitrile (23.6 g, 72%) as a white solid. -114-NMR (400 MHz, DMSO-d6) 6
8.28 (d, J=
1.6 Hz, 1H), 7.88 (dd, J= 8.0, 1.6 Hz, 1H), 7.62 (d, J= 8.0 Hz, 1H), 5.71 (t,
J= 5.6 Hz,
1H), 4.44 (d, J= 5.2 Hz, 2H).
To a solution of 4-(hydroxymethyl)-3-iodo-benzonitrile (11.5 g, 43.5 mmol), 2-
bromo-3,6-difluoro-pyridine (7.08 g, 35.8 mmol) in 1,4-dioxane (80 mL) at 0 C
under
nitrogen, add potassium tert-butoxide (1 M in THF, 43 mL, 43 mmol). Stir the
reaction at
0 C for 1 h, then stir the reaction for 7 h at RT. Dilute the reaction with a
saturated
aqueous solution of NH4C1 (50 mL), then add water (100 mL) and extract with
Et0Ac
(250 mL 3). Combine the organic layers, wash with saturated aqueous NaCl (60
mL),
dry over Na2SO4, filter and concentrate under reduced pressure. Purify the
residue by
silica gel chromatography over using a gradient of 0 to 45% DCM in petroleum
ether to
give the title compound as a white solid (14.28g, 88%). ES-MS m/z 432 (M+H).
Preparation 262
4-[(6-Bromo-5-fluoro-2-pyridyl)oxymethy1]-3-[(E)-2-ethoxyvinyllbenzonitrile
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NC
0 N Br
I
Prepare the title compound essentially as described in Preparation 245 using 4-
[(6-
bromo-5-fluoro-2-pyridyl)oxymethyl]-3-iodo-benzonitrile as starting material
and K2CO3
as the base, heating the reaction at 90 C for 1.5 h. Upon completion,
concentrate the
reaction mixture under reduced pressure, then add water and extract with three
times with
Et0Ac. Combine the organic layers, wash with saturated aqueous NaCl, dry over
Na2SO4,
filter and concentrate under reduced pressure. Purify by silica gel
chromatography using a
gradient of 0 to 60% Et0Ac in petroleum ether to give the title compound as a
white
solid. ES-MS ni/z 377 (M+H).
Preparation 263
4-[(6-Bromo-5-fluoro-2-pyridyl)oxymethyl]-3-(2-oxoethyl)benzonitrile
NC
0 N Br
I
Prepare the title compound essentially as described in Preparation 126 using 4-
[(6-
bromo-5-fluoro-2-pyridyl)oxymethy1]-3-[(E)-2-ethoxyvinyl]benzonitrile,
stirring the
reaction at RT for 20 h. Upon completion, dilute the reaction with water and
extract three
times with Et0Ac. Combine the organic layers, wash with saturated aqueous
NaCl, dry
over Na2SO4, filter and concentrate under reduced pressure to give the title
compound as
a light-yellow solid which is used in Preparation 264 without further
purification. ES-MS
nilz 349, 351 (M+H).
Preparation 264
4-1(6-Bromo-5-fluoro-2-pyridyl)oxymethy11-3-(2-hydroxyethyl)benzonitrile
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NC OH
ONBr
To solution of 4-[(6-bromo-5-fluoro-2-pyridyl)oxymethy1]-3-(2-
oxoethyl)benzonitrile (from Preparation 263, 8.54 g, 22.0 mmol) in Me0H (80
mL) at 0
C, add sodium borohydride (3.59 g, 93.9 mmol) and stir the reaction at RT for
4 h.
Quench the reaction with saturated aqueous solution of NH4C1 and stir for 20
min at RT.
Concentrate the reaction mixture under reduced pressure to remove the solvent.
Dilute the
residue with water (50 mL) and extract with Et0Ac (200 mL < 3). Combine the
organic
layers, wash with saturated aqueous NaC1 (60 mL), dry over Na2SO4, filter and
concentrate under reduced pressure. Purify the residue by silica gel
chromatography using
a gradient of 0 to 30% Et0Ac in petroleum ether to give the title compound as
a yellow
oil (7.1g, 87%). ES-MS nilz 351/353 (M+H).
Preparation 265
Methyl 2-(4-bromo-24(2-(((6-bromo-5-fluoropyridin-2-yl)oxy)methyl)-5-
cyanophenethoxy)methyl)phenyl)acetate
0 0
NC 0
Br
rONB
Prepare the title compound essentially as described in Preparation 224 using
44(6-
bromo-5-fluoro-2-pyridyl)oxymethy11-3-(2-hydroxyethyl)benzonitrile and methyl
2-14-
bromo-2-(bromomethyl)phenyl]acetate as starting materials, using 2,6-di-tert-
buty1-4-
methylpyridine in place of 2,6-di-tert-butylpyridine, and stirring the
reaction at RT
overnight. Upon completion, concentrate the reaction mixture, then add water
and extract
three times with Et0Ac. Combine the organic layers, wash with saturated
aqueous NaC1,
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dry over Na2SO4, filter and concentrate. Purify by silica gel chromatography
using a
gradient of 0 to 100% DCM in petroleum ether to give the title compound as a
pale-
yellow oil. ES-MS m/z 592 (WIT).
Preparation 266
Methyl 2-(54-cyano-2-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
N C 0
0
0 N 0
Prepare the title compound essentially as described in Preparation 216 using
methyl 2-(4-bromo-242-(((6-bromo-5-fluoropyridin-2-yl)oxy)methyl)-5-
cyanophenethoxy)methyl)phenyl)acetate as starting material, using (2-
Dicyclohexylphosphino-2',4',6'-triisopropy1-1,1' -bipheny1)[2-(2'-amino-1, 11-
biphenyl)]palladium(II) methanesulfonate (XPhos Pd G3) as catalyst, and
protecting the
reaction mixture from light during heating. Upon completion, concentrate the
reaction
mixture under reduced pressure to remove the solvent. Dilute the residue with
water and
extract three times with Et0Ac. Combine the organic layers, wash with
saturated aqueous
NaCl, dry over Na2SO4, filter and concentrate under reduced pressure. Purify
by silica gel
chromatography using a gradient of 0 to 33% Et0Ac in petroleum ether to give
the title
compound as a brown solid. ES-MS m/z 433 (M+H).
Preparation 267
2-(54-Cyano-23-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetic acid
N C 0
0 H
0 N 0
,
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Prepare the title compound essentially as described in Preparation 78 using
methyl
2-(54-cyano-23-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetate. Upon completion, quench the reaction with 1 M aqueous citric
acid solution
until pH = 4.5. Filter the crashed solid, wash it with water, collect it and
dry it under
reduced pressure to give the titled compound as a white solid, which is used
in
Preparation 268 without further purification. ES-MS nilz 419 (M+H).
Preparation 268
Methyl (S)-4-(2-(54-cyano-23-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate
/""'
N C 0 H N 0
O¨
N
0
0 N 0 0
0
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-23-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)acetic acid (Preparation 267) and methyl 4-amino-3-(2-methoxyethoxy)-5-
[[(2S)-
oxetan-2-ylimethylamino]benzoate, stirring the reaction at RT overnight. Upon
completion, dilute the reaction with water and extract three times with Et0Ac.
Combine
the organic layers, wash with saturated aqueous NaCl, dry over Na2SO4, filter
and
concentrate under reduced pressure to give the title compound as a pale-brown
oil which
is used in Preparation 269 without further purification. ES-MS nilz 711 (M+H).
Preparation 269
Methyl (S)-2-((54-cyano-23-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate
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NC 0
0 N N 0¨
,
0
0
0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-23-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate (Preparation 268) and 1:1 1,2-dichloroethane : acetic
acid,
heating the reaction at 55 C for 5 h. Upon completion of the reaction, remove
the solvent
under reduced pressure, then add 1:1 Et0Ac : toluene to the residue and
concentrate in
vacuo. Purify by silica gel chromatography using a gradient of 0 to 6% Me0H in
DCM to
give the title compound as an orange oil. ES-MS in/z 693 (M+H).
Preparation 270
Methyl 2-(5-cyano-2-methyl-pheny1)-2,2-difluoro-acetate
N
0 0
Stir 3-i odo-4-methylbenzonitril e (5 g, 19.96 mmol) and copper (12 g, 179.4
mmol) in TEIF (30 mL) and DMSO (80 mL). To this slurry, add methyl
bromodifluoroacetate (6 mL, 51.9 mmol) and continue to stir this mixture at 30
C under
nitrogen for 18 h After this time, add 100 mL of saturated aqueous NaHCO3
solution
followed by 100 mL of Et0Ac. Filter this mixture and wash the solids with
Et0Ac (3 X
50 mL). The filtrate is then separated, and the organic layer washed with
saturated
aqueous NH4C1 solution. Dry the organics over MgSO4, filter, and concentrate.
Purify
this residue via silica gel chromatography (0-25% Et0Ac in hexanes) to give
the product
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as a clear crystalline solid (3.3 g, 73%). 111 NMR (DMSO-d6) 6 2.41 (s, 3H),
3.33 (s,
3H), 7.61 (d, J= 8.0 Hz, 1H), 7.99 (d, J= 7.9 Hz, 1H), 8.04 (d, J= 1.2 Hz,
1H).
Preparation 271
Methyl 2-[2-(bromomethyl)-5-cyano-pheny1]-2,2-difluoro-acetate
Br
N
0 0
Dissolve methyl 2-(5-cyano-2-methyl-pheny1)-2,2-difluoro-acetate (4.4 g, 20
mmol) and N-bromosuccinimide (4 g, 22.47 mmol) in ACN (100 mL). Subject this
solution to a 4100K white lightbulb via flow conditions (1.0 mL/min; 72 feet
of 1/8"
outer diameter reaction tubing, wrapped around a beaker; maintained at 30 C)
twice.
After this processing, concentrate the reaction liquid to dryness and then
purify this
residue via silica gel chromatography (0-10% Et0Ac in hexanes) to give the
product as a
clear, thick oil (3.7 g, 62%). 1H NMR (DMSO-d6) 5 3.41 (s, 3H), 3.89 (s, 2H),
7.61 (d,
J= 7.9 Hz, 1H), 7.99 (d, 8.0 Hz, 1H), 8.03 (s, 1H).
Preparation 272
242-[(6-Bromo-2-pyridyl)oxymethy1]-5-cyano-pheny1]-2,2-difluoro-acetic acid
CN
0
0 H
OF F
Br
Dissolve methyl 242-(bromomethyl)-5-cyano-pheny1]-2,2-difluoro-acetate (6.2 g,
20 mmol) and 2-bromo-6-hydroxypyridine (4.5 g, 2 5 mmol) in DMSO (50 mL). Add
potassium phosphate tribasic (6.6 g, 30 mmol) to this solution and heat to 60
C for 2 h.
After this time, quench the reaction with 1N HC1 (to pH ¨6) and extract with
Et0Ac. Dry
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the combined organics over MgSO4, filter, and concentrate to give the product
as a thick
brown oil (7.8 g, 100%). ES-MS nvz (79Br/81Br) 382.8/384.8 [M+H].
Preparation 273
Methyl 242-[(6-bromo-2-pyridyl)oxymethy1]-5-cyano-pheny1]-2,2-difluoro-acetate
CN
0
CY-
OF F
N
Br
Stir 242-[(6-bromo-2-pyridyl)oxymethyl]-5-cyano-phenyl]-2,2-difluoro-acetic
acid (7.8 g, 20 mmol) in Me0H (100 mL). Add concentrated sulfuric acid (0.1
mL, 2
mmol) and heat to reflux for 30 h. Concentrate the reaction mixture to dryness
and then
purify the residue via silica gel chromatography (0-100% Et0Ac in hexanes) to
give the
product as a white crystalline solid (8 g, 99%). ES-MS tn/z (79Br/81Br)
396.8/398.8
Preparation 274
4-[(6-Bromo-2-pyridyl)oxymethy1]-3-(1,1-difluoro-2-hydroxy-ethyl)benzonitrile
CN
OH
OF F
I
Br
Dissolve methyl 242-[(6-bromo-2-pyridyl)oxymethyl]-5-cyano-phenyl]-2,2-
difluoro-acetate (8 g, 20.14 mmol) in THF (100 mL). To this solution add
lithium
borohydride (0.88 g, 40.4 mmol) and stir at ambient temperature under nitrogen
for 2 h.
After this time, quench the reaction with saturated NH4C1 solution, and
extract with
Et0Ac. Dry the organics over MgS01, filter, and concentrate. Purify this
residue via
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silica gel chromatography (0-50% Et0Ac in hexanes) to give the product as a
thick, clear
oil (4.7 g, 63%). ES-MS nilz (79Br/81Br) 368.8/370.8 [M+Hr.
Preparation 275
342-[(5-Bromo-2-iodo-phenyemethoxy]-1,1-difluoro-ethyl]-446-bromo-2-
pyridyl)oxymethyl]benzonitrile
CN
rp
yx
F F 4111
Br
Br
Dissolve 4-[(6-bromo-2-pyridyl)oxymethy1]-3-(1,1-difluoro-2-hydroxy-
ethypbenzonitrile (4.5 g, 12 mmol) in THE (60 mL) and DMF (10 mL). To this
solution
add sodium hydride (0.6 g, 15 mmol; 60% mass in mineral oil) and stir at
ambient
temperature under nitrogen for 5 min. Then add 4-bromo-2-(chloromethyl)-1-
iodobenzene (4.8 g, 14 mmol) and continue stirring at ambient temperature
under
nitrogen for 18 h. After this time, quench the reaction with saturated aqueous
NH4C1
solution, and extract with Et0Ac. Dry the organics over MgSO4, filter, and
concentrate
Purify this residue via silica gel chromatography (0-30% Et0Ac in hexanes) to
give the
product as a thick clear oil (3.9 g, 48%) ES-MS rn/z (79Br/81Br) 663.0/665.0
[M+H]
Preparation 276
Ethyl 2[4-bromo-2[[242-[(6-bromo-2-pyridypoxym ethy1]-5-cyano-pheny1]-2,2-
difluoro-ethoxy]methyl]phenyl]acetate
CN
0
0
OF F
Br
I
Br
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Dissolve 342-[(5-bromo-2-iodo-phenyl)methoxy1-1,1-difluoro-ethy11-4-[(6-
bromo-2-pyridyl)oxymethyl]benzonitrile (3.9 g, 5.9 mmol) and [(4,5-
bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2'-amino-1,11-bi phenyl)]pal 1
adium (II)
methanesulfonate (XantPhos Pd G3, 0.6 g, 0.6 mmol) in Tiff' (30 mL). To this
solution
add (2-ethoxy-2-oxo-ethyl)zinc bromide (0.5 M in ether) (18 mL, 9.0 mmol) and
heat this
mixture to 60 C under nitrogen for 18 h. After this time, cool the reaction
to ambient
temperature, quench the reaction with saturated aqueous NaHCO3 solution, and
extract
with Et0Ac. Dry the organics over MgSO4, filter, and concentrate. Purify this
residue
via silica gel chromatography (0-30% Et0Ac in hexanes) to give the product as
a thick,
light brown oil (1.6 g, 44%). ES-MS m/z (79Br/"Br) 623.2/625.2 [M-F1-1]+.
Preparation 277
Ethyl 2-(54-cyano-6,6-difluro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzeneacyclonaphane-14-yl)acetate
F F
NC 0
0
I
Dissolve ethyl 2-[4-bromo-2-[[242-[(6-bromo-2-pyridyl)oxymethy1]-5-cyano-
pheny1]-2,2-difluoro-ethoxy]methyl]phenyl]acetate (1.6 g, 2.9 mmol) in 1,4-
dioxane (30
mL). To this solution add KOAc (0.64 g, 6.39 mmol) and bis(pinacolato)diboron
(0.8 g,
3.09 mmol) and bubble nitrogen through this for 10 min. After this time add
1,1'-
bis(diphenylphosphino)ferrocene-palladium(II)dichloride DCM complex (0.11 g,
0.132
mmol) and heat this mixture to 80 C under nitrogen for 18 h. After this time,
cool the
reaction to ambient temperature, dilute with saturated aqueous NaCl solution,
and extract
with Et0Ac. Dry the organics over MgSO4, filter, and concentrate. Dissolve
this residue
in 1,4-dioxane (50 mL) and water (3 mL) and add potassium phosphate tribasic
(1.4 g, 6.5
mmol) followed by chloro(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-
bipheny1)[2-
(2'-amino-1 ,11-biphenyl)]palladium(II) (XPhos Pd G2, 0.1 g, 0.125 mmol). Heat
this
mixture to 60 C for 2 h. After this time, cool the reaction to ambient
temperature,
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quench the reaction with saturated aqueous NH4C1 solution, and extract with
Et0Ac. Dry
the organics over MgSO4, filter, and concentrate. Purify this residue via
silica gel
chromatography (0-50% Et0Ac in hexanes) to give the product as a white solid
(249 mg,
21%). ES-MS (m/z) 465.2 (M+H).
Preparation 278
Methyl (S)-24(54-cyano-6,6-difluoro-3.8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclonaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxatan-2-ylmethyl)-
1H-
benzokilimidazole-6-carboxylate
F F
=C
NCIIr
0 )
1 0
0
I 0¨
0
0¨
Dissolve ethyl 2-(54-cyano-6,6-difluro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzeneacyclonaphane-14-yl)acetate (249 mg, 0.54 mmol) in ACN (5 mL), TI-IF
(1.8
mL) and water (1.8 mL). To this solution add 1,5,7-triazabicyclo[4.4.0]dec-5-
ene (0.23 g,
1.62 mmol) and stir this mixture at ambient temperature for 2 h. After this
time, quench
the reaction with saturated aqueous NH4C1 solution and extract with Et0Ac. Dry
the
organics over MgSO4, filter, and concentrate. Dissolve this residue in DMF (2
mL) and
add methyl 4-amino-3-(2-methoxyethoxy)-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate
(0.055 g, 0.18 mmol), DMF (0.09 mL, 0.5 mmol) and HATU (0.09 g, 0.24 mmol) and
stir
at ambient temperature for 18 h. After this time, quench the reaction with
saturated
aqueous NH4C1 solution, and extract with Et0Ac. Dry the organics over MgSO4,
filter,
and concentrate. Dissolve this residue in 1,2-dichloroethane (1 mL), add
acetic acid (1
mL), and heat this mixture to 50 C for 18 h. Concentrate the mixture to
dryness and
purify the residue via silica gel chromatography (0-100% Et0Ac in hexanes) to
give the
title compound as an off-white solid (61 mg, 52.5%). ES-MS (m/z) 711.4 (M+H).
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Preparation 279
4[(2-Chloropyrimidin-4-yl)oxymethyd-3-iodo-benzonitrile
NC
0 N CI
IN
To a solution of 2-chloropyrimidin-4-ol (8.50 g, 65.1 mmol) in DMF (150 mL),
add Cs2CO3 (42.5 g, 130 mmol), and 4-(bromomethyl)-3-iodo-benzonitrile (21.04
g,
65.35 mmol). Stir the reaction mixture at ambient temperature for 16 h. Pour
the crude
reaction into water and collect the precipitate via filtration. Dissolve the
solid material
into DCM and wash twice with water. Dry the organic phase over MgSO4, filter,
and
concentrate under reduced pressure to give the title compound (20.3 g, 84%) as
a light
orange solid, which is used in Preparation 280 without further purification.
ES/MS m/z
372 (M+H).
Preparation 280
4[(2-Chloropyrimidin-4-yl)oxymethyl]-3-[(E)-2-ethoxyvinyl]benzonitrile
NC
0 N CI
N
To a solution of 4-[(2-chloropyrimidin-4-yl)oxymethyl]-3-iodo-benzonitrile
(10.1
g, 27.2 mmol) in THF (150 mL), add tribasic potassium phosphate (40 mL, 80
mmol, 2 M
solution in water), 2-[(E)-2-ethoxyviny1]-4,4,5,5-tetramethy1-1,3,2-
dioxaborolane (7.5
mL, 35 mmol), and bis(triphenylphosphine)palladium(II) dichloride (953 mg,
1.36
mmol). Sparge the solution with nitrogen for 15 min, then stir at 55 C for 4
h. Dilute the
reaction mixture with water and extract with Et0Ac. Dry the organic phase with
MgSO4,
filter, and concentrate under reduced pressure. Dissolve the residue in Et0Ac
and wash
with water to remove pinacol. Dry the organic phase with MgSO4, filter, and
concentrate
under reduced pressure. Purify the residue via silica gel chromatography using
a gradient
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of 0 to 30% Et0Ac in hexanes to give the title compound (5.97 g, 70%). ES/MS
in/z
(35C1/37C1) 315/317 [M+Hr.
Preparation 281
4-[(2-Chloropyrimidin-4-yl)oxymethyl]-3-(2-hydroxyethyl)benzonitrile
NC
11101
0 N CI
I 'r
To a solution of 4-[(2-chloropyrimidin-4-yl)oxymethyl]-3-[(E)-2-
ethoxyvinyl]benzonitrile (5.75 g, 18.2 mmol) in THF (85 mL), add hydrochloric
acid (46
mL, 184 mmol, 4M solution in dioxane). Stir at ambient temperature for 2.5 h.
Concentrate the reaction under reduced pressure, then dilute the residue with
DCM.
Adjust the mixture to pH = 8 using saturated aqueous NaHCO3 solution, then
extract with
DCM. Dry the organic phase with MgSO4, filter, and concentrate under reduced
pressure.
Dissolve the residue in Me0H (100 mL), cool to 0 C in an ice bath, then
gradually add
sodium borohydride (1.28 g, 33.7 mmol). Stir the mixture at 0 C for 30 min.
Quench the
reaction mixture with 1 M aqueous NaOH, then further dilute the solution with
water and
DCM, and extract with DCM. Dry the organic phase over MgSO4, filter, and
concentrate
under reduced pressure. Purify the residue via silica gel chromatography using
a gradient
of 0 to 50% Et0Ac in hexanes give the title compound (2.86 g, 44%). ES/MS m,/z
(35C1/37C1) 289/291 [M-FFI]t
Preparation 282
Methyl 244-bromo-24242-[(2-chloropyrimidin-4-yl)oxymethyl]-5-cyano-
phenyljethoxymethyliphenyljacetate
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CO2Me
NC 0 14111
Br
N C
Cool a solution of 4-[(2-chloropyrimidin-4-yl)oxymethy1]-3-(2-
hydroxyethyl)benzonitrile (200 mg, 0.69 mmol) in anhydrous DCM (4.6 mL) to 0
C in
an ice bath, add methyl 2[4-bromo-2-(bromomethyl)phenyl]acetate (556 mg, 1.73
5 mmol), then add 2,6-di-tert-butylpyridine (0.31 mL, 1.38 mmol) and silver
trifluoromethanesulfonate (354 mg, 1.38 mmol). Stir the mixture for 1 h at 0
C, then
allow to stir for 16 h at ambient temperature. Filter the reaction mixture
through a pad of
diatomaceous earth and rinse the pad with DCM. Concentrate the filtrate under
reduced
pressure and purify the residue via silica gel chromatography using a gradient
of 0 to 40%
10 Et0Ac in hexanes to give the title compound (128 mg, 35%). ES/MS in/z
(79Br/51Br)
529/531 [M+H]t
Preparation 283
Methyl 2424242-[(2-chloropyrimidin-4-yl)oxymethyl]-5-cyano-
phenyl]ethoxymethy11-
4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-yl)phenyl]acetate
CO2Me
NC 0 el 0
B-176
0
ONCI
I
To a solution of methyl 2-[4-bromo-2-[2-[2-[(2-chloropyrimidin-4-yl)oxymethy1]-
5-cyano-phenyflethoxymethyl]phenyl]acetate (566 mg, 0.715 mmol) in 1,4-dioxane
(7.5
mL), add bis(pinacolato)diboron (222 mg, 0.86 mmol), KOAc (177 mg, 1.80 mmol),
and
1,1' -bis(diphenylphosphino)ferrocene palladium(II) dichloride dichloromethane
complex
(30 mg, 0.036 mmol). Sparge the solution with nitrogen for 10 min, then heat
to 80 'C.
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Stir for 19 h. Cool the reaction to ambient temperature and filter through a
plug of silica
gel, washing the silica gel plug with DCM. Concentrate the filtrate under
reduced
pressure to obtain the title compound, which is used in Preparation 284
without further
purification. ES/MS nilz (35C1/37C1) 496/498 [M+H] (as boronic acid).
Preparation 284
Methyl 2-(54-cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetate
NC 0
CO2Me
N
N
To a solution of methyl 2-[24242-[(2-chloropyrimidin-4-yl)oxymethyl]-5-cyano-
phenyl]ethoxymethyl]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-
yl)phenyl]acetate
(Preparation 283, 130 mg, 0.225 mmol) in THF (5.6 mL), add tribasic potassium
phosphate (0.68 mL, 0.68 mmol, 1 M aqueous solution). Sparge the solution with
nitrogen for 10 min, then add chloro(2-dicyclohexylphosphino-2',4',6'-
triisopropy1-1,1'-
bipheny1)[2-(2'-amino-1,1'-biphenyl)]pa1ladium(H) (XPhos Pd G2, 8.9 mg, 0.011
mmol)
and resume sparging for a further 5 min. Heat the solution at 50 C for 6 h.
Cool the
reaction to ambient temperature, then add water and extract with Et0Ac. Dry
the
combined organic phases over MgSO4, filter, and concentrate under reduced
pressure.
Purify the crude residue via silica gel chromatography using a 0 to 40%
gradient of
Et0Ac in hexanes to give the title compound (21 mg, 22%). ES/MS nilz 416
(M+H).
Preparation 285
2-(54-Cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-dibenzenacyclononaphane-
14-
ypacetic acid
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NC
CO2H
0 N
Prepare the title compound essentially as described in Preparation 217 using
methyl 2-(54-cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-ypacetate, stirring the reaction at ambient temperature for 2 h. Upon
completion,
quench the reaction with 5% aqueous citric acid bringing the pH to 4, then
dilute mixture
with Et0Ac. Separate the layers and extract the aqueous layer with Et0Ac. Wash
the
combined organic phases with saturated aqueous NaCl solution. Dry the organic
phase
over MgSO4, filter, and concentrate under vacuum. Triturate the resulting
solid with
water to give the title compound. ES/MS in/z 402 (M+H).
Preparation 286
Methyl (S)-4-(2-(54-cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate
NC 0
H
0 N 0 lel
0 CO2 Me
N
Prepare the title compound essentially as described in Preparation 164 using 2-
(54-
cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic
acid and methyl 4-amino-3-methoxy-5-[[(2S)-oxetan-2-yl]methylamino]benzoate as
starting materials, adding 2,4,6-tripropy1-1,3,5,2,4,6-trioxatriphosphorinane-
2,4,6-trioxide
as a 50% solution in DMI'm and heating the solution to 42 C. for 24 h. Cool
reaction to
ambient temperature and then add water. Extract the aqueous layer with Et0Ac,
then
wash the organics with saturated aqueous NaHCO3 solution. Dry the organics
over
MgSO4, filter, and concentrate under reduced pressure to give the title
compound which
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is used in Preparation 287 without further purification (23.6 mg, 83%). ES/MS
in/z 650
(M+H),
Preparation 287
Methyl (S)-24.54-cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yOmethyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
0
,,===0
t 0
I / 0
0 N
0-
0
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-methoxy-5-((oxetan-2-
ylmethyl)amino)benzoate (Preparation 286) and using a 5:1 mixture of 1,2-
dichloroethane
: acetic acid as solvent, heating the reaction to 53 C for 22 h. Upon
completion, cool the
reaction mixture to ambient temperature, dilute with DCM, and concentrate
under
reduced pressure. Dissolve the residue in Et0Ac and concentrate under reduced
pressure
twice. Purify the residue via silica gel chromatography using a gradient of 0
to 5% Me0H
in DCM to give the title compound. ES/MS 111/Z 632 (M+H).
Preparation 288
Ethyl 4-[(E)-2-ethoxyviny1]-6-(trifluoromethyppyridine-3-carboxylate
F
NR, I
0
To a solution of ethyl 4-chloro-6-(trifluoromethyl)nicotinate (15 g, 57.4
mmol)
and Cs2CO3 (37 g, 112.4 mmol) in 1,4-dioxane (130 mL) and water (45 mL), add 2-
[(E)-
2-ethoxyviny1]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (13.4 mL, 62.9 mmol),
then add
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tetrakis(triphenylphosphine)palladium(0) (7 g, 5.75 mmol). Sparge the solution
with
nitrogen, then heat the mixture to 90 C and stir for 16 h. Concentrate the
reaction
mixture under reduced pressure, then dilute with water. Extract aqueous layer
with
Et0Ac, dry the combined organics over Na2SO4, filter, and concentrate under
reduced
pressure. Purify the residue silica gel chromatography eluting with a gradient
of 0 to 23%
Et0Ac in petroleum ether to give the title compound (14.5 g, 84%). ES/MS m/z
290
(MII-1).
Preparation 289
[4-[(E)-2-Ethoxyviny1]-6-(trifluoromethyl)-3-pyridyl]methanol
I
N
0 H
To a solution of ethyl 4-[(E)-2-ethoxyviny1]-6-(trifluoromethyl)pyridine-3-
carboxylate (14.5 g, 48.1 mmol) in THF (100 mL) at -78 C under an inert
atmosphere,
add diisobutylaluminum hydride (150 mL, 150 mmol, 1M solution in toluene).
Remove
the reaction vessel from the cooling bath and allow to stir at ambient
temperature for 5 h.
Quench the reaction by adding a saturated aqueous solution of potassium sodium
tartrate
(200 mL) at 0 C, then add DCM (100 mL). Extract the mixture with DCM, then
wash the
combined organic phases with saturated aqueous NaCl solution. Dry the organic
phase
over MgSO4, filter, and concentrate under reduced pressure. Purify the residue
via silica
gel chromatography eluting with a gradient of 0 to 30% Et0Ac in petroleum
ether to give
the title compound (11 g, 90%). ES/MS m/z 247 (Mt).
Preparation 290
5-[(6-Bromo-2-pyridyl)oxymethyl]-4-[(E)-2-ethoxyviny1]-2-
(trifluoromethyppyridine
F
N I
0 rN B
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To a solution of [44(E)-2-ethoxyviny11-6-(trifiuoromethy1)-3-pyridyllmethanol
(10.5 g, 40.4 mmol) and 2-bromo-6-fluoroyridine (5.4 mL, 51 mmol) in 1,4-
dioxane (100
mL), add potassium tert-butoxide (52 mL, 52 mmol, 1M solution in TT-IF) at 0
C. Stir the
reaction at 0 C for 30 min, then at ambient temperature for 2 h. Quench the
reaction with
saturated aqueous NH4C1 solution (100 mL), then extract with Et0Ac. Dry the
organics
over MgSO4, filter, and concentrate under reduced pressure. Purify the residue
via silica
gel chromatography using a gradient of 0 to 30% Et0Ac in petroleum ether to
give the
title compound (17 g, 96%). ES/MS m/z (79Br/81Br) 402/404 [M-hfi].
Preparation 291
245-[(6-Bromo-2-pyridyl)oxymethy1]-2-(trifluoromethyl)-4-pyridyl]ethanol
F0 H
N I
0 N Br
To a solution of 5-[(6-bromo-2-pyridypoxymethy1]-4-[(E)-2-ethoxyvinyl]-2-
(trifluoromethyppyridine (16.5 g, 39.3 mmol) in THF (300 mL) and water (500
mL) add
mercuric acetate (14.45 g, 44.89 mmol) at 0 C. Stir the mixture for 30 min at
0 C, then
add 50% aqueous K2CO3 solution (210 mL) and sodium borohydride (6.31 g, 165
mmol).
Stir the mixture at 0 C for 2.5 h. Filter the reaction mixture, washing the
filter with
Et0Ac. Transfer the filtrate to a separatory funnel and extract with Et0Ac.
Dry the
combined organic phases with Na2SO4, filter, and concentrate under reduced
pressure.
Purify the residue via silica gel chromatography using a gradient of 0 to 25%
Et0Ac in
petroleum ether to give the title compound (12.1 g, 81%). ES/MS rn/z
(79Br/81Br) 376/378
[M+Hr
Preparation 292
442-[(5-Bromo-2-chloro-phenyl)methoxy]ethyl]-54(6-bromo-2-pyridyl)oxymethyl]-2-
(trifluoromethyl)pyridine
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CI
B r
0 rN B
To a solution 245-[(6-bromo-2-pyridyl)oxymethy1]-2-(trifluoromethyl)-4-
pyridyllethanol (5.46 g, 14.3 mmol), 4-bromo-1-chloro-2-(chloromethyl)benzene
(5.19 g,
21.4 mmol), and 2,6-di-tert-butyl-4-methylpyridine (6 g, 28.64 mmol) in 1,2-
dichloroethane (100 mL), add silver trifluoromethanesulfonate (12 g, 46.24
mmol) at
ambient temperature under an inert atmosphere. Heat the reaction to 70 C and
stir for 18
h. Concentrate the crude reaction under reduced pressure, then dilute the
resultant residue
with water. Extract from the aqueous with Et0Ac, then wash the combined
organic
phases with saturated aqueous NaCl solution. Dry the organic phase over
Na2SO4, filter,
and concentrate under reduced pressure. Purify the residue via silica gel
chromatography
eluting with a gradient of 0 to 20% Et0Ac in petroleum ether to give the title
compound
(3.3 g, 37%). ES/MS nilz 580 (M+H).
Preparation 293
14-Chloro-56-(trifluoromethyl)-3,8-dioxa-2(2,6),5(3,4)-dipyridina-1(1,3)-
benzenacyclononaphane
N I C I
0 N
I
Prepare the title compound essentially as described in Preparation 216 using 4-
[2-
[(5-bromo-2-chloro-phenyl)methoxy]ethy1]-5-[(6-bromo-2-pyridypoxymethyl]-2-
(trifluoromethyppyridine as starting material and using (2-
dicyclohexylphosphino-2',4',6'-
triisopropy1-1,1'-bipheny1)[2-(2'-amino-1,11-biphenyl)]palladium(II)
methanesulfonate
(XPhos Pd G3) as catalyst, heating the reaction at 110 C for 15 h.
Concentrate the crude
reaction under reduced pressure, then dilute the residue with water. Extract
from the
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aqueous with Et0Ac, then wash the combined organic phases with saturated
aqueous
sodium chloride solution. Dry the organic phase over Na2SO4, filter, and
concentrate
under reduced pressure. Purify the residue via reverse phase HPLC
chromatography using
a gradient of 30-100% ACN in aqueous formic acid to give the title compound
(403 mg,
14%) as a white solid. ES/MS m/z 421 (M+H).
Preparation 294
14-(4,4,5,5-Tetramethy1-1,3,2-dioxaborolan-2-y1)-56-(trifluoromethyl)-3,8-
dioxa-
2(2,6),5(3,4)-dipyridina-1(1,3)-benzenacyclononaphane
Fo
9
N., I
0 N
I
To a solution of 14-chloro-56-(trifluoromethyl)-3,8-dioxa-2(2,6),5(3,4)-
dipyridina-
1(1,3)-benzenacyclononaphane (275 mg, 0.62 mmol), KOAc (155 mg, 1.56 mmol),
and
bis(pinacolato)diboron (321 mg, 1.24 mmol) in 1,4-dioxane (6.5 mL), add
chloro(2-
dicyclohexylphosphino-2',41,6'-triisopropy1-1,11-bipheny1)[2-(2'-amino-1,1'-
biphenyMpalladium(II) (XPhos Pd (12, 50 mg, 0.06 mmol). Sparge the solution
with
nitrogen, then heat to 80 C and stir for 18 h. Concentrate the reaction
mixture under
reduced pressure, then dilute the residue with water. Extract from the aqueous
with
Et0Ac, then wash the combined organic phases with saturated aqueous NaC1
solution.
Dry the organic phase over Na2SO4, filter, and concentrate under reduced
pressure. Purify
the residue via silica gel chromatography eluting with a gradient of 0 to 30%
Et0Ac in
petroleum ether to give the title compound (274 mg, 70%). ES/MS m/z 513 (M+H).
Preparation 295
Methyl 2-(chloromethyl)-7-(2-methoxyethoxy)-3-[[(28)-oxetan-2-
yl]methylThenzimidazole-5-carboxylate
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r". 0
CI-Mr N
O¨
N 10,
0
0
0
To a solution of methyl 4-amino-3-(2-methoxyethoxy)-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate (310 mg, 0.93 mmol) in Et0H (6 mL), add 2-chloro-1,1,1-
trimethoxyethane (710 mg, 4.50 mmol) at ambient temperature. Heat the mixture
to 90 C
for 2 h, then concentrate the reaction under reduced pressure. Purify the
residue via silica
gel chromatography eluting with a gradient of 0 to 5% Me0H in DCM to give the
title
compound (360 mg, 94%). ES/MS m/z 369 (M+H).
Preparation 296
Methyl (S)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-2-((56-(trifluoromethyl)-
3,8-
dioxa-2(2,6),5(3,4)-dipyridina-1(1,3)-benzenacyclononaphane-14-yOmethyl)-1H-
benzokilimidazole-6-carboxylate
N., I
0--
0 N N
0
0
0
To a solution of 14-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)-56-
(trifluoromethyl)-3,8-dioxa-2(2,6),5(3,4)-dipyridina-1(1,3)-
benzenacyclononaphane (160
mg, 0.25 mmol, 81 wt%), methyl 2-(chloromethyl)-7-(2-methoxyethoxy)-3-[[(28)-
oxetan-
2-yl]methyl]benzimidazole-5-carboxylate (256 mg, 0.62 mmol, 89 mass %), and
tribasic
potassium phosphate (172 mg, 0.79 mmol) in 2-methyltetrahydrofuran (1.6 mL)
and
water (0.32 mL), add chloro(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1 -
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bipheny1)12-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos Pd G2, 26 mg, 0.03
mmol).
Sparge the solution with nitrogen, then heat the reaction to 80 C for 18 h.
Concentrate
the reaction mixture under reduced pressure, then dilute the residue with
water. Extract
the mixture with Et0Ac, then wash the combined organic phases with saturated
aqueous
NaCl solution. Dry the organic phase over Na2SO4, filter, and concentrate
under reduced
pressure. Purify the residue via silica gel chromatography eluting with a
gradient of 0 to
60% Et0Ac in petroleum ether to obtain the title compound (100 mg, 53%). ES/MS
m/z
719 (M+H).
Preparation 297
Ethyl 2-(2-methyloxazol-4-yl)acetate
0
)\-0
Add ethyl 4-chloro-3-oxo-butanoate (40 g, 243 mmol) to a solution of acetamide
(15 g, 254 mmol) in toluene (80 mL) at RT. Heat to 130 C and stir for 12 h.
Filter the
reaction mixture and concentrate the filtrate under reduced pressure. Purify
the residue
via silica gel chromatography eluting with a gradient of 10 to 15% Et0Ac in
petroleum
ether to give the title compound as a yellow oil with 30% purity (19 g, 13%).
ES/MS nilz
170 (M+H)
Preparation 298
2-(2-Methyloxazol-4-ypethanol
OH
re\-=
)-0
Add calcium chloride (26 g, 234 mmol) to a solution of ethyl 2-(2-methyloxazol-
4-yl)acetate (20 g, 35.5 mmol) in Et0H (400 mL). Stir the mixture for 20 min,
cool to 0
C, and then add sodium borohydride (10 g, 265 mmol). Stir the reaction mixture
at RT
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for 12 h and then adjust the pH to 7 with 1 M aqueous HC1. Extract the mixture
with
DCM (3 x 200 mL), dry the combined organics over Na9SO4, filter, and
concentrate.
Purify the reside via silica gel chromatography eluting with a gradient of 50
to 100%
Et0Ac in petroleum ether, followed by a gradient of 0 to 15% Me0H in DCM to
give the
title compound as a yellow oil with 70% purity (5 g, 78%). ES/MS m/z 128
(M+H).
Preparation 299
Methyl 3-fluoro-5-hydroxy-4-nitro-benzoate
02N
0
HO
o
At -40 C, add boron tribromide (2 M solution in DCM, 65.4 mL, 131 mmol)
dropwise to a solution of methyl 3-fluoro-5-methoxy-4-nitro-benzoate (30 g,
131 mmol)
in DCM (500 mL). Stir the mixture at -40 C for 30 min and then at ambient
temperature
for 16 h. Pour the mixture into ice water (2 L) and extract with DCM (3 x 800
mL).
Wash the combined organic layers with saturated aqueous NaCl solution (2 x 300
mL),
dry over Na2SO4, filter, and concentrate. Purify the residue via silica gel
chromatography
eluting with a gradient of 0 to 30% Et0Ac in petroleum ether to give the title
compound
as a yellow solid (16.5 g, 59%). 'HNIVIR (CDC13) 6 10.31 (s, 1H), 7.63 (s,
1H), 7.41 (d,
1H, J = 11), 3.98 (s, 3H). '9F NMR (CDC13) 6 -113.37 (s).
Preparation 300
Methyl 3-fluoro-542-(2-methyloxazol-4-ypethoxy]-4-nitro-benzoate
02 N
0
0
N A's=
)¨ 0
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-230-
At 0 C, add diispropylazodicarboxylate (5.6 g, 28 mmol) to a solution of
methyl
3-fluoro-5-hydroxy-4-nitro-benzoate (3 g, 13.9 mmol) and 2-(2-methyloxazol-4-
ypethanol (3 g, 16.5 mmol) in THF (30 mL). Allow the mixture to warm to
ambient
temperature and stir for 12 h. Dilute the mixture with water (30 mL) and
extract with
Et0Ac (3 x 50 mL). Wash the combined organics with saturated aqueous NaCl (100
mL), dry over Na2SO4, filter, and concentrate. Purify the residue via silica
gel
chromatography eluting with a gradient of 0 to 50% Et0Ac in petroleum ether to
give the
title compound as a light yellow solid with 17% purity (6 g, 23%). ES/MS nilz
325
(M+H).
Preparation 301
Methyl 3-[2-(2-methyloxazol-4-ypethoxy]-4-nitro-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate
HN"'"'"-r-1
02N
0
Add (S)-oxetan-2-ylmethanamine (15 g, 6.29 mmol) and TEA (1 mL, 7.17 mmol)
to a solution of methyl 3-fluoro-542-(2-methyloxazol-4-ypethoxy1-4-nitro-
benzoate (6 g,
3.15 mmol) in DMSO (150 mL). Stir the mixture at 80 C for 12 h, cool to
ambient
temperature, dilute with water (300 mL), and extract with Et0Ac (3 < 400 mL).
Wash
the combined organic layers with saturated aqueous NaCl solution (3 x 500 mL),
dry over
Na2SO4, filter, and concentrate. Purify the residue via silica gel
chromatography eluting
with a gradient of 0 to 60% Et0Ac in petroleum ether to afford the title
compound as a
yellow oil with 24% purity (2 g, 39%). ES/MS nilz 392 (M+H).
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Preparation 302
Methyl 4-amino-342-(2-methyloxazol-4-yl)ethoxy]-5-[[(2S)-oxetan-2-
yl]m ethyl amino]benzoate
H2N
0
N
)-0
Add palladium on carbon (1 g, lOwt% Pd) to a solution of methyl 3-[2-(2-
methyloxazol-4-yl)ethoxy]-4-nitro-5-[[(2S)-oxetan-2-yl]methylamino]benzoate (2
g, 1.23
mmol) in Et0Ac (150 mL). Purge the reaction vessel with hydrogen gas and
evacuate
three times. Attach a balloon of hydrogen gas to the vessel and stir for 2 h.
Filter the
mixture and concentrate under reduced pressure. Purify the residue by silica
gel
chromatography eluting with a gradient of 0 to 100% Et0Ac in petroleum ether,
and then
purify further by preparative HPLC [column: Phenomenex C18 75 40 mm, 3 u.m;
mobile phase: 18 to 48% ACN in aqueous NHIHCO3 (10 mM)] to give the title
compound as a white solid (88 mg, 19%). ES/MS m/z 362 (M+H).
Preparation 303
Methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-(2-methyloxazol-4-yl)ethoxy)-5-
((oxetan-2-ylmethyl)amino)benzoate
NC
4111/ 0
0
0 ,N HO
1411 o
0
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-232-
Prepare the title compound essentially as described in Preparation 86 using
methyl
4-amino-342-(2-methyloxazol-4-ypethoxy]-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate
(80 mg, 0.215 mmol) and 2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(l,2)-
dibenzenacyclononaphane-14-ypacetic acid, and the product is used without
further
purification in Preparation 304. ES/MS m/z 744 (M+H).
Preparation 304
Methyl (S)-2-054-eyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yOmethyl)-4-(242-methyloxazol-4-ypethoxy)-1-(oxetan-
2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylate
NC
4
0 .. .0 10
NT'
0
0 N N
0-
0
NO
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacy cl ononaphane-14-ypacetamido)-3-(2-(2-methyloxazol-4-yl)ethoxy)-5-
((oxetan-2-ylmethyl)amino)benzoate (Preparation 303) and using a 1:1 mixture
of 1,2-
dichloroethane : acetic acid. Upon completion, cool the reaction to ambient
temperature
and remove volatiles under vacuum. Add 1:1 Et0Ac/toluene and concentrate three
times
to remove residual acetic acid. Purify the residue via silica gel
chromatography eluting
with a gradient of 0 to 6% Me0H in DCM to give the title compound as an orange
solid.
ES/MS miz 726 (M+H).
Preparation 305
tert-Butyl N-(2-hydroxyethyl)-N-(2,2,2-trifluoroethyl)carbamate
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-23 3 -
0
HO¨/
FK
F F
A mixture of 2-bromoethanol (5 g, 38.8 mmol) and 2,2,2-trifluoroethylamine
(9.66 g, 97.0 mmol) in ethanol (50 mL) was heated at 60 'V for 3 h in a sealed
tube in a
microwave reactor, then the reaction mixture was concentrated. To the residue
was added
TEA (5.2 mL, 3.7 g, 37 mmol) and di-tert-butyl pyrocarbonate (5.6 mL, 5.3 g,
24 mmol)
and the mixture was stirred overnight at 55 'V under N2. The reaction mixture
was
partitioned between Et0Ac and water, and the organic phase was concentrated
under
reduced pressure. The residue was purified by silica gel chromatography using
a gradient
of 0 to 38% Et0Ac in petroleum ether to give the title compound (2.5 g, 50%)
as a yellow
oil. 1H NMR (400 MHz, DMSO) 6 4.76 (m, 1H), 4.06 (q, J= 9.5 Hz, 2H), 3.67 (m,
2H),
3.30 (q, J= 6 Hz, 2H), 1.41 (s, 9H).
Preparation 306
Methyl 312-[tert-butoxycarbony1(2,2,2-trifluoroethyl)amino]ethoxy]-5-fluoro-4-
nitro-
benzoate
0
0
02N 0
ONO
F>r)
To a solution of methyl 3-fluoro-5-hydroxy-4-nitro-benzoate (1.5 g, 6.6 mmol),
tert-butyl N-(2-hydroxyethyl)-N-(2,2,2-trifluoroethyl)carbamate (2.1 g, 7.8
mmol) and
triphenylphosphine (3.5 g, 13 mmol) in THF (10 mL), add DIAD (2.8 mL, 13 mmol)
at 0
C. After the addition, stir the mixture at 20 C for 18 h. Dilute mixture with
water (50
mL) and extract with Et0Ac (3 x 50 mL). Dry the combined organic layers over
Na2SO4,
filter, and concentrate under reduced pressure. Purify by silica gel
chromatography
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eluting with a gradient of 0 to 20% Et0Ac/petroleum ether to give the title
compound as a
yellow oil (3.2 g, 93%). ES/MS miz 340.9 (M-Boc+H).
Preparation 307
Methyl 3-[2-[tert-butoxycarbony1(2,2,2-trifluoroethyl)amino]ethoxy]-4-nitro-5-
[[(2S)-
oxetan-2-yl]methylamino]benzoate
0
d.. rq'
40)
02N 0
FF
F >1)
Prepare the title compound essentially as described in Preparation 301 using
methyl 3-[2-[tert-butoxycarbony1(2,2,2-trifluoroethyl)amino]ethoxy]-5-fluoro-4-
nitro-
benzoate and fumaric acid;(S)-oxetane-2-ylmethanamine, heating the reaction at
100 C
for 3 h in a microwave reactor. ES/MS nilz 508.1 (M+H).
Preparation 308
Methyl 4-amino-342-[tert-butoxy carbony1(2,2,2-trifluoroethyl)amino]ellioxy]-5-
[[(2S)-
oxetan-2-yl]methylamino]benzoate
d., 0
H2N 0
N
EF
To a solution of methyl 342-[tert-butoxycarbony1(2,2,2-
trifluoroethyl)amino]ethoxy]-4-nitro-5-[[(2S)-oxetan-2-yl]methylamino]benzoate
(1.1 g,
1.8 mmol) in Me0H (10 mL), add Pd/C (810 mg, 10 mass%). Stir the mixture under
hydrogen (15 psi) for 2 h at ambient temperature. Add diatomaceous earth,
filter, and
concentrate the filtrate under reduced pressure. Purify the residue by silica
gel
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-23 5 -
chromatography eluting with a gradient of 0 to 9% Me0H in DCM to give the
title
compound (940 mg, >99%). ES/MS iv/z 478.1 (M+H).
Preparation 309
Methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-342-[tert-butoxycarbonyl(2,2,2-
trifluoroethypamino]ethoxy]- 5-[[(2S)-oxetan-2-yl]methylamino]benzoate
NC
14111 0
0
HNN--A
0 NJ}U HN 00
,
0 1411
0
F
N0
0<
Prepare the title compound essentially as described in Preparation 86 using
2454-
cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetic acid
and methyl 4-amino-342-[tert-butoxycarbony1(2,2,2-trifluoroethypamino]ethoxy]-
5-
[[(2S)-oxetan-2-yl]methylamino]benzoate. ES/MS rtilz 860.2 (M+H).
Preparation 310
Methyl (S)-4-(2-((tert-butoxycarbonyl)(2,2,2-trifluoroethyl)amino)ethoxy)-2-
((54-cyano-
3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-
1-
(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carb oxylate
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-23 6-
NC
1111 0
d""Q
0
, 0
,
¨
/ 0
F
Prepare the title compound essentially as described in Preparation 102 using
methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-342-[tert-butoxycarbony1(2,2,2-
trifluoroethyl)amino]ethoxy]- 5-[[(2S)-oxetan-2-yl]methylamino]benzoate and a
mixture
of 1:1 DCM : acetic acid as solvent, heating the reaction at 55 C for 18 h.
Upon
completion, cool the reaction to ambient temperature, dilute with toluene, and
concentrate
under reduced pressure. Purify the residue by silica gel chromatography
eluting with a
gradient of 0 to 14% Me0H in DCM to give the title compound as a brown oil.
ES/MS
nilz 842.2 (M+H).
Preparation 311
(S)-4-(2-((tert-Butoxycarbonyl)(2,2,2-trifluoroethypamino)ethoxy)-2-454-cyano-
3,9-
dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-
(oxetan-2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
NC
0 ,==
Ni"
0
0 N N
OH
0
F
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-237-
Prepare the title compound essentially as described in Example 5 using methyl
(S)-4-(2-((tert-butoxycarbonyl)(2,2,2-trifluoroethyl)amino)ethoxy)-2-((54-
cyano-3,9-
di oxa-2(2,6)-pyri dina-1(1,3),5(1,2)-dibenzenacycl ononaphane-14-yl)methyl)-1-
(oxetan-2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylate 1,5,7-triazabicyclo[4.4.0]dec-5-
ene in a
mixture of 3:1:1 ACN THF : water, heating the reaction at 55 C for 2 h. Upon
completion, cool the reaction to ambient temperature and quench with 1 M
aqueous
formic acid to give a precipitate. Filter and collect the precipitate to give
the title
compound, which is used without further purification in Example 54. ES/MS nilz
828.2
(M+H).
Preparation 312
2-Chloro-6-[(4-chloro-2-iodo-phenypmethoxy]pyridine
CI
1411 0 N CI
Prepare a solution of 4-chloro-2-iodo-1-methyl -benzene (40 g, 158 mmol) and N-
bromosuccinimide (27.3 g, 153 mmol) in ACN (500 mL). Pump through a
photochemical
reactor consisting of a coiled PFA reaction tube (1/8" outer diameter, 53 mL
volume)
around a 4100K white lightbulb (42W, 150W equivalent), with a flowrate of 1.3
mL/min
and maintaining the system temperature between 30 and 40 C. Once complete,
pump
ACN (100 mL) through the reactor at the same rate. Drip the output into a
flask
containing a suspension of 6-chloropyridin-2-ol (21.8 g, 168 mmol) and K2CO3
(44.1 g,
319 mmol) in ACN (400 mL). After full addition of the brominated material,
stir at RT
for 1 h. Filter the crude reaction mixture and concentrate under reduced
pressure. Purify
the residue via silica gel chromatography using 0 to 10% Et0Ac in hexanes to
give 45.8 g
of the title compound (76%). ES-MS in/z 380 and 382 (M+H).
Preparation 313
2-Chloro-6-[[4-chloro-2-[2-ethoxyvinyl]phenyl]methoxy]pyridine
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CI
N C
To a solution of 2-chloro-6-[(4-chloro-2-iodo-phenyl)methoxy]pyridine (5.0 g,
13
mmol) in THF (70 mL) under N2 add aqueous potassium phosphate (2 M, 150 mL,
300
mmol) and 1-ethoxyethene-2-boronic acid pinacol ester (1/1.3 isomeric mixture,
3.81 mL,
5 17.1 mmol) and stir the mixture for 5 min. Add
bis(triphenylphosphine)palladium(II)
dichloride (470 mg, 0.663 mmol) and heat the mixture to 65 C for 21 h. Cool
the
mixture to RI and add water (100 mL) and Et0Ac (100 mL). Stir the mixture for
5 min at
RT, separate the organic layer and extract the aqueous layer with Et0Ac (2><
100 mL).
Combined the organic layers and wash with water (100 mL), saturated aqueous
NaC1
10 (100 mL). Concentrate the organics and purify the residue by filtration
through a plug of
silica gel using DCM eluant to give the title compound as a brown solid (4.37
g, 100%).
ES/MS nilz 324 (M+H).
Preparation 314
245-Chloro-246-chloro-2-pyridyl)oxymethyl]phenyl]acetaldehyde
CI
'0
0
6,1
CI
To a solution of 2-chloro-64[4-chloro-242-ethoxyvinyl]phenyl]methoxy]pyridine
(54.2 g, 161.1 mmol) in acetone (525 mL) and water (175 mL) add hydrochloric
acid (35
mL, 407 mmol, 36.5% in water) and heat to 65 C for one hour. Cool the
reaction to RI
and dilute with water (250 mL) and MTBE (250 mL). Separate the organic phase
and
extract the aqueous with MTBE (3 x 250 mL). Wash the combined organic layers
with
2M aqueous Na2CO3 (250 mL), water (250 mL), saturated aqueous NaCl (250 mT,),
dry
over Na2SO4 and remove the solvents in vacuo to give 44.5 g of the title
compound (93%)
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which is used in Preparation 315 without further purification. ES-MS in/z
296.0, 298.0
(M+H).
Preparation 315
2-[5-Chloro-2-[(6-chloro-2-pyridypoxymethyl]phenyl]ethanol
CI
0 H
0
C I
To a solution of 245-chloro-2-[(6-chloro-2-
pyridyl)oxymethyl]phenyl]acetaldehyde (Preparation 314, 44.5 g, 150.3 mmol) in
Et0H
(400 mL) at 5 C add sodium borohydride (11.85 g, 300.7 mmol) portionwise and
stir at
RI for one hour. Cool the reaction to 0 C and quench with water (250 mL) and
adjust
the pH to 6 with 5% aqueous citric acid. Add MTBE (400 mL), separate the
organic
phase and extract the aqueous phase with MTBE (3 >< 250 mL). Combine the
organic
layers and wash with water (250 mL), saturated aqueous NaCl (250 mL), dry over
Na2SO4 and the remove the solvents in vacuo. Purify the crude material by
silica gel
chromatography using a gradient of 20 to 40% Et0Ac in cyclohexane to give
31.95 g of
the title compound (71%) which crystallizes upon standing ES-MS nilz 2980,
300.0
(M+H).
Preparation 316
24[242-[(5-Bromo-2-iodo-phenyl)methoxy]ethyl]-4-chloro-phenyl]methoxy]-6-
chloro-
pyridine
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C,
Br
0
o
CI
To a solution of 245-chloro-2-[(6-chloro-2-pyridyl)oxymethyl]phenyl]ethanol
(31.80 g, 106.6 mmol) in THF (360 mL) at 0 C add sodium hydride (60 wt% in
mineral
oil, 8.14 g, 203 mmol) and stir for 20 min. To this add 4-bromo-2-
(bromomethyl)-1-iodo-
benzene (49.4 g, 131 mmol) portionwise and allow the mixture to warm to RT and
stirred
for 18 h. To this add more sodium hydride (60 wt /0 in mineral oil, 1.25g,
31.3 mmol)
followed by more 4-bromo-2-(bromomethyl)-1-iodo-benzene (4.9 g, 13 mmol) and
stir
the mixture at RT for another 5 h. Cool the reaction to 0 C and quench with
5% aqueous
citric acid (250 mL) and add MTBE (200 mL). Separate the organic phase and
extract the
aqueous with MTBE (3 > 250 mL). Combine the organic layers and wash with water
(200
mL), saturated aqueous NaCl (200 mL), dry over Na2SO4 and remove the solvents
in
vacuo. Purify the residue by silica gel chromatography using a gradient of 40
to 100%
DCM in cyclohexane to give 45.05 g of the title compound (71%) as a yellow
oil. ES-
MS nilz 593.8 (M+H).
Preparation 317
Ethyl 2-[4-bromo-2-[2-[5-chloro-2-[(6-chloro-2-
pyridyl)oxymethyl]phenyl]ethoxymethyl]phenyl]acetate
CI 0
0
Br
o
"- CI
Stir a solution of 24[242-[(5-bromo-2-iodo-phenyOmethoxy]ethyl]-4-chloro-
phenyl]methoxy]-6-chloro-pyridine (13.11 g, 22.1 mmol), and chloro[(4,5-
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bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2'-amino-1,1'-
biphenyl)Thalladium(II)]
(XantPhos Pd G2, 1100g. 1.107 mmol) in THF (50 mL) at RT under N2. To this
mixture add bromo-(2-ethoxy-2-oxo-ethyl)zinc (0.4 M in THF, 103 mL, 40 mmol)
dropwise and heat the mixture to 65 C for 3 h. Cool the reaction mixture to 0
C and
quench with 5% aqueous citric acid (200 mL) and add MTBE (200 mL). Separate
the
organic phase and extract the aqueous phase with MTBE (3 100 mL). Wash the
combined organic layers with water (200 mL), saturated aqueous NaC1 (200 mL),
dry
over Na2SO4 and remove the solvents under vacuum. Purify the residue by silica
gel
chromatography using a gradient of 20 to 100% DCM in cyclohexane to give 9.15
g of
the title compound (75%) as a dark brown oil. ES-MS nilz 552.0, 554.0 (1VI+H).
Preparation 318
Ethyl 2-12-1245-chloro-2-[(6-chloro-2-pyridyl)oxymethyl]phenyllethoxymethyl]-4-
(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)phenyl]acetate
CI 0
0
0
N 0'6'0
cI
Bubble N2 gas through a solution of ethyl 2-14-bromo-2-1245-chloro-2-[(6-
chloro-
2-pyridypoxymethyl]phenyl]ethoxymethyl]phenyl]acetate (7.50g, 13.6 mmol),
bis(pinacolato)diboron (4.39 g, 16.9 mmol), and KOAc (4.1 g, 41 mmol) in
dioxane (270
mL) for 15 min. To this mixture add
dichlorobis(tricyclohexylphosphine)palladium (II)
(1.02 g, 1.35 mmol) and heat to 90 C for 20 h. To this mixture add additional
bis(pinacolato)diboron (0.439 g, 1.69 mmol),
dichlorobis(tricyclohexylphosphine)palladium (II) (0.10 g, 0.13 mmol), and
KOAc
(0.410g, 4.1 mmol). Heat the mixture to 90 C for 4 h. Cool the mixture to RI
and filter
through a plug of silica gel eluting with DCM to give 9.11 g of the title
compound as a
crude dark brown oil (112%), which is used in Preparation 319 without further
purification. ES-MS m/z 600.2, 602.2 (M+H).
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Preparation 319
Ethyl 2-(54-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)acetate
CI 0
0,
0 N 0
,
Bubble N2 gas through a solution of ethyl 2-[2-[2-[5-chloro-2-[(6-chloro-2-
pyridyl)oxymethyl]phenyl]ethoxymethy1]-4-(4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-
yl)phenyl]acetate (9.10g, 15.2 mmol) and aqueous potassium phosphate tribasic
(1 M, 91
mL, 91 mmol) in THF (610 mL) for 15 min. To this mixture add [chloro(2-
dicyclohexylphosphino-2',41,6'-tri-i-propyl -1,11-biphenyl)(21-amino-1,11-
bipheny1-2-y1)
palladium(II)] (XPhos-Pd-G2, 1.22 g, 1.52 mmol) and heat to 65 C for one
hour. Cool
the reaction to RT, dilute with water (300 mL) and extract with MTBE (3 x 250
mL).
Wash the combined organics with water (250 mL), saturated aqueous NaCl (250
mL), dry
over MgSO4, filter, and concentrate in vacuo. Purify the residue by silica gel
chromatography using a gradient of 80 to 100% DCM in cyclohexane to give 1.6 g
of the
title compound (24.1%) as a white solid. ES-MS nilz 438.2 (M+H).
Preparation 320
2-(54-Chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-
14-
yl)acetic acid
CI 0
0 H
0 N 0
To a solution of ethyl 2-(54-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate (900 mg, 2.06 mmol) in ACN (5 mL), 1,4-
dioxane (5 mL), and water (5 mL) add 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-
a]pyrimidine (500 mg, 3.52 mmol) and heat to 50 "V for 3 h. Cool the reaction
to RT,
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quench with 1N HC1 (2 mL), dilute with water and extract with Et0Ac (2 x 250
mL).
Combine the organics, wash with saturated aqueous NaC1, dry over MgSO4, filter
and
concentrate in vacuo to give 835 mg of the title compound (99%) as a light
yellow solid
ES-MS tr//z 410.9 (M+H).
Preparation 321
Methyl 342-[tert-butyl(dimethyl)silyl]oxyethoxy]-5-fluoro-4-nitro-benzoate
0
0
02N
0
I
To a 0 C solution of methyl 3-fluoro-5-hydroxy-4-nitro-benzoate (2 g, 9.3
mmol), 2-((tert-butyldimethylsilyl)oxy)ethanol (2.6 g, 14 mmol) and
triphenylphenylphosphine (3.5 g, 13 mmol) in THF (30 mL), add DIAD (3.8 g, 19
mmol).
Allow to warm to 20 C and stir for 12 h. Dilute with water (50 mL) and
extract with
Et0Ac (3 x 80 mL). Wash the combined organic layers with saturated aqueous
sodium
chloride solution (100 mL), dry over Na2SO4, filter, and concentrate under
reduced
pressure. Purify the residue by silica gel chromatography eluting with 0-20%
Et0Ac/petroleum ether to give the title compound as colorless oil (4 g, 92%).
1H NM_R
(CDC13) 6 7.59 (s, 1H), 7.51 (d, J = 24 Hz, 1H), 4.25 (t, J= 5 Hz, 2H), 3.98
(t, J = 5 Hz,
2H), 3.97 (s, 3H), 0.88 (s, 9H), 0.07 (s, 6H).
Preparation 322
Methyl 3-(2-hydroxyethoxy)-4-nitro-5-[[(2S)-oxetan-2-yl]methylamino]benzoate
0
H
02N
H
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To a solution of methyl 342-[tert-butyl(dimethyl)silyl]oxyethoxy1-5-fluoro-4-
nitro-benzoate (2 g, 4.32 mmol) in DMSO (200 mL), add TEA (2.5 mL, 18 mmol)
and
(S)-oxetane-2-ylmethanamine (21 g, 8.7 mmol). Stir at 80 C for 12 h. Dilute
with water
(200 mL) and extract with Et0Ac (3 x 300 mL). Wash the combined organic layers
with
saturated aqueous sodium chloride solution (400 mL), dry over Na2SO4, filter,
and
concentrate in vacuo. Purify the residue by silica gel chromatography eluting
with 0-50%
Et0Ac/petroleum ether to give the title compound (320 mg, 18%). 1H NMR (CDC13)
6
7.16 (s, 1H), 6.93 (s, 1H), 6.39 (s, 1H), 5.11 (m, 1H), 4.72 (m, 1H), 4.59 (m,
1H), 4.24 (t,
j = 4 Hz, 2H), 3.97 ¨ 3.93 (m, 5H), 3.50 (t, j = 4 Hz, 2H), 2.73 (m, 1H), 2.58
(m, 1H).
Preparation 323
Methyl 3-12-[tert-butyl(dimethypsilylioxyethoxy]-4-nitro-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate
0
HN
O2N
Si
I
To a solution of methyl 3-(2-hydroxyethoxy)-4-nitro-5-[[(2S)-oxetan-2-
yl]methylamino]benzoate (320 mg, 0.78 mmol) in DCM (10 mL), add tert-
butyldimethylchlorosilane (200 mg, 1.29 mmol) and imidazole (130 mg, 1.90
mmol). Stir
the mixture at 20 C for 2 h. Dilute with water (20 mL) and extract with DCM
(3 x 30
mL). Wash the combined organic layers with saturated aqueous sodium chloride
solution
(50 mL), dry over Na2SO4, filter, and concentrate under reduced pressure.
Purify the
residue by silica gel chromatography eluting with 0-20% Et0Ac/petroleum ether
to give
the title compound (400 mg, 93%). ES/MS (m/z): 441.1 (M+H).
Preparation 324
Methyl 4-amino-342-[tert-butyl(dimethyl)silylioxyethoxy]-5-[[(2S)-oxetan-2-
yl]m ethyl aminoThenzoate
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0
H N C:o"
H2N
o
I
To a solution of methyl 342-[tert-butyl(dimethypsilyl]oxyethoxyl-4-nitro-5-
[[(2S)-oxetan-2-ylimethylamino]benzoate (670 mg, 1.22 mmol) in Et0Ac (100 mL),
add
Pd/C (300 mg, 10 mass%). Purge with hydrogen gas three times and stir at 20 C
for one
hour under a balloon filled with hydrogen. Filter the mixture and concentrate
the filtrate
under reduced pressure. Purify the residue by silica gel chromatography
eluting with 0-
10% Me0H/DCM to give the title compound (475 mg, 95%). ES/MS (m/z): 411.4
(M+H).
Preparation 325
Methyl (S)-342-[tert-butyl(dimethyl)silyl]oxyethoxy]-4-(2-(54-chloro-3,8-dioxa-
2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetamido)-5-((oxetan-2-
ylmethypamino)benzoate
CI leiII
0
H H
0 0 Oil 0
0
0
0 I
' Si
To a solution of 2-(54-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetic acid (80 mg, 0.20 mmol) and methyl 4-amino-
3-
[2-[tert-butyl(dimethyl)silyl]oxyethoxy]-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate (87
mg, 0.21 mmol) in DMF (3 mL), add DMA (0.07 mL, 0.4 mmol) followed by HATU
(124 mg, 0.32 mmol). Stir the mixture at 20 C for 2 h, then dilute with water
(20 mL) to
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form a solid. Filter off the solid and dry under vacuum to give the title
compound (132
mg, 84%). ES/MS rn/z (35C1/37C1) 802.4/804.4 [M+11].
Preparation 326
Methyl (S)-442-[tert-butyl(dimethypsilyl]oxyethoxy]-2-454-chloro-3,8-dioxa-
2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate
CI
/ 0
0 N
0¨
I
0
/
0-Si
Heat a solution of methyl (S)-342-[tert-butyl(dimethyl)silyl]oxyethoxy]-4-(2-
(54-
chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)acetamido)-5-((oxetan-2-ylmethyl)amino)benzoate (132 mg, 0.165 mmol) in
acetic
acid (3 mL) to 80 C for 1.5 h, then cool to ambient temperature and
concentrate under
reduced pressure. Purify the residue by silica gel chromatography eluting with
a gradient
of 50 to 100% Et0Ac/toluene followed by 2 to 5% Me0H/Et0Ac to give the title
compound (62 mg, 48%). The product elutes at 1.8 min by LC-ES/MS but does not
ionize. Conditions for LCMS analysis: 2 50 Xbridge C18 3.5 litm at 50 C, 5-
95%
ACN in 10 mM aqueous NH4HCO3 (pH 9) over 1.5 minutes, then 95% ACN for 0.5
min.
Preparation 327
Methyl 3-[2-(dimethylamino)ethoxy]-5-fluoro-4-nitro-benzoate
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F
02N 0
0 0,
N,
To a solution of methyl 3-fluoro-5-hydroxy-4-nitro-benzoate (2 g, 9.30 mmol)
in
THF (20 mL), add N,N-dimethylethanolamine (994 mg, 11.15 mmol),
triphenylphosphine
(2.92 g, 1 L 15 mmol) and DIAD (2.25 g, 1 L15 mmol). Stir reaction at ambient
temperature for 18 h. Filter the reaction mixture and concentrate the filtrate
liquids under
reduced pressure. Purify the residue via silica gel flash chromatography
eluting with a
gradient of 0 to 10% Me0H in DCM to give the title compound as a yellow oil
(690 mg,
26%). ES/MS in/z 287 (M+H).
Preparation 328
Methyl 342-(dimethylamino)ethoxy1-4-nitro-5-[[(2S)-oxetan-2-
ylimethylamino]benzoate
02N)0
0 0,
N,
To a solution of methyl 3-12-(dimethylamino)ethoxy1-5-fluoro-4-nitro-benzoate
(690 mg, 2.41 mmol) in DMSO (8 mL), add (25)-oxetan-2-yl-methanamine (420 mg,
4.8
mmol) and TEA (1.46 g, 14.4 mmol). Stir the reaction mixture at 80 C for 18
h. Cool the
mixture to ambient temperature, dilute with water (50 mL) and extract with
Et0Ac (2 X
50 mL). Combine the organics, wash with water and saturated aqueous NaCl
solution (2 X
50 mL), dry over Na2SO4, filter and evaporate under reduced pressure. Purify
the residue
via silica gel flash chromatography eluting with a gradient of 0 to 20% Me0H
in DCM to
give the title compound as a colorless oil (1.03 g, 72%). ES/MS in/z 354
(M+H).
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Preparation 329
Methyl 4-amino-342-(dimethylamino)ethoxy]-54[(2S)-oxetan-2-
yl]m ethyl aminoThenzoate
H
H 2 N 0
0 0,
N-
To a solution of methyl 342-(dimethylamino)ethoxy]-4-nitro-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate (1.03 g, 1.75 mmol) in Et0Ac (100 mL), add 10% Pd/C
(200
mg, 0.18 mmol). Stir the mixture under hydrogen gas (14.5 psi) at ambient
temperature
for 2 h, then filter through diatomaceous earth and concentrate the filtrate
under reduced
pressure. Purify the residue by preparative HPLC to give the title compound as
a white
solid (102 mg, 17%). [column:Welch Xtimate C18 75 x 40mm, 3 Jim; Mobile phase
A:
I-190 with NH3H2O+NH4HCO3; Mobile phase B: ACN; Gradient: B from 22% to 52%).
ES/MS m/z 324 (M+H).
Preparation 330
Methyl (S)-4-(2-(54-chloro-3 ,8-dioxa-2(2,6)-pyridina-1(1,3), 5(1,2)-
dib enzenacycl ononaphane-14-yl)acetamido)-3 -(2 -(dimethylamino)ethoxy)-5-
((oxetan-2-
ylmethyl)amino)benzoate
ço
CI 0 NH
ON)0 lel 0
0
I
0
To a solution of 2-(54-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid (75 mg, 0.18 mmol) and methyl 4-
amino-3-
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[2-(dimethylamino)ethoxy]-5-11(2S)-oxetan-2-yllmethylaminoThenzoate (60 mg,
0.18
mmol) in DMF (2 mL), add HATU (115 mg, 0.3 mmol) and DIPEA (0.10 mL, 0.5
mmol). Stir the mixture at ambient temperature for 17 h. Quench the reaction
with water
and filter the white solid that precipitates. Concentrate the filtrate liquids
and extract with
Et0Ac. Combine the organic layers, dry the organics over MgSO4, filter, and
concentrate
to give a solid residue. Combine the first white solid with the residue to
give the title
compound as a white solid (173 mg, 132%). ES/MS m/z 715 (M+H).
Preparation 331
Methyl (S)-24(54-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-(dimethylamino)ethoxy)-1-(oxetan-2-
ylmethyl)-1H-benzokflimidazole-6-carboxylate
CI
/ 0
0 Isk., N
I 0
0
N ¨
/
Heat a mixture of methyl (S)-4-(2-(54-chloro-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetamido)-3-(2-
(dimethylamino)ethoxy)-
5-((oxetan-2-ylmethyl)amino)benzoate (130 mg; 0.18 mmol) in acetic acid (2 mL)
and
1,2-dichloroethane (1.5 mL) at 55 C for 21 h. Cool the mixture to ambient
temperature
and concentrate under reduced pressure. Purify the residue via silica gel
flash
chromatography eluting with a gradient of 0 to 10% Me0H in DCM to give the
title
compound as a white solid (89 mg, 70%). ES/MS nilz 697 (M+H).
Preparation 332
Methyl 342-ttert-butoxycarbonyl(methypamino]ethoxy]-5-fluoro-4-nitro-benzoate
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00
0 0,
0
To a solution of methyl 3-fluoro-5-hydroxy-4-nitro-benzoate (2 g, 9.30 mmol)
and
tert-butyl-(2-hydroxyethyl)methyl carbamate (3.43 g, 18.6 mmol) in THF (20
mL), add
triphenylphosphine (3.7 g, 14 mmol) and DIAD (2.82 g, 13.9 mmol). Stir
reaction at
ambient temperature for 12 h. Concentrate the mixture under reduced pressure
and purify
the residue via silica gel chromatography eluting with a gradient of 0 to 20%
Et0Ac in
petroleum ether to give the title compound as a yellow oil (2.3 g, 66%). ES/MS
(m/z): 373
(M+1-1).
Preparation 333
Methyl 342-[tert-butoxycarbonyl(methyl)amino]ethoxy]-4-nitro-5-[[(2S)-oxetan-2-
yl]methylamino]benzoate
.... <-"\
HN sO"'
02N 0
0 0,
0
To a solution of methyl 342-[tert-butoxyearbonyl(methyl)amino]ethoxy]-5-
fluoro-4-nitro-benzoate (1 g, 2.68 mmol) in DMSO (60 mL), add a 3.6% w/w
solution of
(2S)-oxetan-2-yl-methanmine in Et0H (13.0g, 5.4 mmol) and TEA (1.6 g, 16
mmol). Stir
the reaction mixture at 80 C for 12 h. Cool the mixture to ambient
temperature, dilute
with water (100 mL) and extract with Et0Ac (3 x 100 mL). Combine the organics,
wash
with water and saturated aqueous NaCl solution (3 x 100 mL), dry over Na2SO4,
filter,
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and concentrate under reduced pressure. Purify the residue via silica gel
chromatography
eluting with a gradient of 0 to 20% Et0Ac in petroleum ether to give the title
compound
as a yellow oil (750 mg, 62%). ES/MS (m/z): 440 (M+1-1-Boc).
Preparation 334
Methyl 4-amino-3-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]-5-[[(2S)-oxetan-
2-
yl]methylamino]benzoate
ço
N H
H2N
0
0
0
N 0
y
To a solution of methyl 3-[2-[tert-butoxycarb onyl(methyl)amino]ethoxy]-4-
nitro-
5-11(2S)-oxetan-2-yl]methylaminolbenzoate (700 mg, 1.59 mmol) in Et0Ac (50
mL), add
10% Pd/C (200 mg, 0.18 mmol). Stir the mixture under hydrogen gas (15 psi) at
ambient
temperature for 2 h. Filter the reaction mixture through diatomaceous earth
and
concentrate the filtrate under reduced pressure to give the title compound
(536 mg, 82%).
ES/MS (m/z): 410 (M+1-1)
Preparation 335
Methyl (S)-3-(2-((tert-butoxycarbonyl)(methypamino)ethoxy)-4-(2-(54-chloro-3,8-
dioxa-
2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-y1)acetamido)-5-
((oxetan-2-
ylmethyl)amino)benzoate
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CI 0
0 N 0 lel 0
0
N 0
y
0
To a solution of 2-(54-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetic acid (80 mg, 0.19 mmol) and methyl 4-amino-
3-
[2-[tert-b utoxycarbonyl(methyl)amino]ethoxy]-5-[[(2S)-oxetan-2-
yl]methylaminoThenzoate (82 mg, 0.19 mmol) in DMF (2 mL), add HATU (130 mg,
0.33
mmol) and DIEA (0.11 mL, 0.62 mmol). Stir the mixture at ambient temperature
for 15 h,
then quench with water and extract with Et0Ac. Dry the organics over MgSO4,
filter, and
concentrate under reduced pressure to give the title compound as a solid (155
mg, 80%),
which is used in Preparation 336 without further purification. ES/MS en /z):
801 (MI--I).
Preparation 336
Methyl (S)-4-(2-((tert-butoxycarbonyl)(methyl)amino)ethoxy)-24(54-chloro-3,8-
dioxa-
2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-
2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylate
CI 0
C.)
= 0
0 N
I 0 ¨
0
0
N
/ 0 _____________________________________________________________
Heat a mixture of methyl (S)-3-(2-((tert-butoxycarbonyl)(methyl)amino)ethoxy)-
4-(2-(54-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
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-25 3 -
yl)acetamido)-5-((oxetan-2-ylmethyl)amino)benzoate (Preparation 335, 155 mg,
0.19
mmol) in acetic acid (1.5 mL) and 1,2-dichloroethane (1.5 mL) at 55 C for 18
h. Cool
the reaction to ambient temperature and concentrate under reduced pressure.
Purify the
residue via silica gel chromatography eluting with a gradient of 0 to 10% Me0H
in DCM
to give the title compound as a white solid (150 mg, 99%). ES/MS (m/z): 783
(M+H).
Preparation 337
(S)-4-(2-(((eri-Butoxycarbonyl)(methypamino)ethoxy)-2-054-chloro-3,8-dioxa-
2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid
CI
Ni"-=
/ 0
0 N N =
,
0 H
0
0
/
To a nitrogen degassed mixture of methyl (S)-4-(2-((tert-
butoxycarbonyl)(methyl)amino)ethoxy)-2-((54-chloro-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate (150 mg, 0.19 mmol) in ACN (2 mL), 1,4-dioxane
(2
mL), and water (0.4 mL), add 1,5,7-triazabicyclo[4.4.0]dec-5-ene (86 mg, 0.60
mmol)
and heat the mixture to 55 C for 36 h. Cool the reaction mixture to ambient
temperature,
concentrate under reduced pressure, and purify the residue via reverse-phase
chromatography using a gradient of 10 to 90% ACN in water (0.1% formic acid
added to
both solvents) to give the title compound as a white solid (64 mg, 43%). ES/MS
(m/z):
769 (M+H).
Preparation 338
Ethyl 2-(4-bromo-5-fluoro-2-methyl-phenyl)acetate
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Me
0,-
0
Br
To solution of 1-bromo-2-fluoro-4-iodo-5-methyl-benzene (11 g, 34 mmol) and
chloro[(4,5-bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2'-amino-1,1'-
biphenyl)Thalladium(II) (XantPhos-Pd-G2, 3.5 mg, 3.4 mmol) in THE (45 mL), add
(2-
ethoxy-2-oxoethyl)zinc(II) bromide (0.50 M in THE, 110 mL, 60 mmol) at RT and
stir
the mixture at 65 C overnight under N2. Quench the reaction by addition of IN
HC1 (100
mL), then extract the mixture with Et0Ac (3 x 100 mL). Combine the organic
layers and
wash with aqueous K2CO3 (100 mL), dry over Na2SO4 and concentrate in vacuo.
Purify
the residue by silica gel chromatography using a gradient of 0 to 20% Et0Ac in
petroleum ether to give the title compound (9.73 g, 98%) as a pale-yellow oil.
ES/MS
m/z 274.8 (M+H)
Preparation 339
Ethyl 2-[4-bromo-2-(bromomethyl)-5-fluoro-phenyl]acetate
Br
0,
Br 0
Prepare a solution of ethyl 2-(4-bromo-5-fluoro-2-methyl-phenyl)acetate (5.9
g,
21 mmol) and N-bromosuccinimide (3.6 g, 20 mmol) in ACN (110 mL). Pump the
solution through a photochemical reactor consisting of a coiled PFA reaction
tube (1/8"
outer diameter, 53 mL volume) around a 4100K white lightbulb (42W, 150W
equivalent),
with a flowratc of 1.3 mL/min and maintaining the system temperature between
30 and
40 C. Once complete, pump ACN (100 mL) through the reactor at the same rate.
Dilute
the output mixture with water (500 mL) and heptane (200 mL) Extract the
aqueous layer
with heptane (200 mL) and wash the combined organic layers with saturated
aqueous
sodium thiosulfate solution (2 x 100 mL) Dry the organic layer over MgSO4,
filter, and
concentrate under vacuum. Purify the residue via silica gel chromatography
using a
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gradient of 0 to 10% Et0Ac in hexanes to give 7.0 g of the title compound (92%
yield,
90% pure). 1H-NMR (400 MHz, CDC13) 6 7.59 (d, .1 = 7 Hz, 1H), 7.09 (d, .1 = 9
Hz, 1H),
4.50 (s, 2H), 4.20 (q, J= 7.5 Hz, 2H), 3.75 (s, 2H), 1.29 (t, 1= 7.5 Hz, 3H).
Preparation 340
Ethyl 244-bromo-242454(6-chloro-2-pyridyl)oxymethyl]-2-cyano-
phenyllethoxymethyl]-5-fluoro-phenyl]acetate
Br
=
I I
0
0
0
CI
Charge a reaction vessel with 4-[(6-chloro-2-pyridyl)oxymethyl]-2-(2-
hydroxyethyl)benzonitrile (700 mg, 2.4 mm ol), ethyl 244-bromo-2-(bromomethyl)-
5-
fluoro-phenyllacetate (1.3 g, 3.2 mmol, 88 mass%), anhydrous DCM (20 mL) and
2,6-di-
tert-butylpyridine (835 ittL, 3.61 mmol). Cool the mixture to 0 C under
nitrogen
atmosphere and add silver trifluoromethanesulfonate (878 mg, 3.38 mmol). Allow
the
mixture to reach RT and stir overnight. Filter the reaction mixture through
Celite ,
concentrate the filtrate and purify the residue by silica gel chromatography
using a
gradient of 20 to 100% DCM in cyclohexane to give the title compound (369 mg,
67 wt%
pure, 18%) as a colorless waxy solid. ES-MS m/z 561, 563, 565 (M-F11).
Preparation 341
Ethyl 2-[2-[2-[5-[(6-chloro-2-pyridyl)oxymethy1]-2-cyano-phenyflethoxymethyl]-
5-
fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]acetate
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0
0
0
0
1
CI
Charge a reaction vessel with ethyl 2-[4-bromo-2-[2-[5-[(6-chloro-2-
pyridyl)oxymethy1]-2-cyano-phenyllethoxymethyl]-5-fluoro-phenyllacetate (340
mg,
0.41 mmol, 67 wt% pure), KOAc (140 mg, 1.40 mmol), bis(pinacolato)diboron (190
mg,
0.73 mmol) and anhydrous 1,4-dioxane (4 mL). Bubble the mixture with nitrogen,
add
dichlorobis(tricyclohexylphosphine)palladium(II) (61 mg, 0.08 lmmol) and stir
at 90 C
in a preheated bath for 5 h. Cool the reaction mixture to RT, add saturated
aqueous
NaHCO3 and Et0Ac and filter the mixture through Celite'. Separate aqueous
layer and
wash the organic layer with water and saturated aqueous NaCl, dry over Na2SO4,
filter
and concentrate. Purify the residue via silica gel chromatography using a
gradient of 0 to
100% Et0Ac in DCM to give the title compound (275 mg, 75 wt% pure, 84%) as a
yellow waxy solid. ES-MS nilz 609 and 611 (M+H).
Preparation 342
Ethyl 2-(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)acetate
I I
0
4111
0 N 0
F
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Charge a reaction vessel with ethyl 2424245-[(6-chloro-2-pyridyl)oxymethy1]-2-
cyano-phenyl]ethoxymethyl]-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-
2-
yl)phenyl]acetate (190 mg, 0.23 mmol, 75 wt% pure) and TI-IF (7.8 mL). Bubble
the
mixture with nitrogen, then add potassium phosphate tribasic (1.0 M in water,
1.2 mL, 1.2
mmol) and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-
bipheny1)[2-(2'-
amino-1,1'-biphenyl)]palladium(H) (XPhos Pd G2, 20 mg, 0.025 mmol). Stir the
mixture
at 70 'V in a preheated bath for 30 min. Cool the reaction mixture to RT and
add water
and Et0Ac. Separate the aqueous layer and wash the organic layer with water
and
saturated aqueous NaCl, dry over Na2SO4, filter and remove solvent. Purify the
residue
via silica gel chromatography using a gradient of 0 to 5% Et0Ac in DCM to
provide the
title compound (49 mg, 90 wt% pure, 42%) as a pale brown solid. ES-MS m/z 447
(M+H).
Preparation 343
2 -(54-Cyano-16-fluoro-3 , 8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-
yl)acetic acid
I
0
0 H
0 N 0
F
Add 1,5,7-triazabicyclo[4.4.0]dec-5-ene (62 mg, 0.44 mmol) to a suspension of
ethyl 2-(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)acetate (72 mg, 0.145 mmol, 90 mass%) in a
mixture of
ACN (1.5 mL), THF (500 1..tL) and water (500 L). Purge the mixture with N2
and stir at
50 C for 15 min. Remove the organic solvents with a nitrogen stream. Add
Et0Ac and
aqueous citric acid (5%). Separate the aqueous layer and wash the organic
layer with
water and saturated aqueous NaC1, dry over Na2SO4, filter and remove the
solvent to
provide the title compound (65mg, 97%, 91 mass%) as a white solid. ES-MS nilz
419
(M+H).
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Preparation 344
Methyl (S)-2-((54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl )methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylm
ethyl )-
1H-benzo[d]imidazole-6-carboxylate
I I
r-o
L-1
0 N N 0
0 ¨
0
0
To a solution of 2-(54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)acetic acid (65 mg, 0.14 mmol, 91 mass%) and
methyl 4-
amino-3-(2-methoxyethoxy)-5-[[(2S)-oxetan-2-yl]methylamino]benzoate (45 mg,
0.145
mmol) in anhydrous DMF (1.4 mL) under nitrogen atmosphere, add HATU (70 mg,
0.18
mmol) and DIPEA (74 [iL, 0.42 mmol). Stir the mixture at RT for 3 h and dilute
with
water and Et0Ac. Separate the aqueous layer and wash the organic layer with
water and
saturated aqueous NaC1, dry over Na2SO4, filter and remove solvent. Dissolve
the residue
in 1,2-dichloroethane (0.9 mL) and acetic acid (0.9 mL) and heat the mixture
at 60 C
under N2 for 8 h. Cool the reaction mixture to RT, concentrate solvents under
reduced
pressure, dry under vacuum at 35-40 C and purify the residue via silica gel
chromatography using a gradient of 25 to 100% Et0Ac in DCM to provide the
title
compound (50 mg, 49%) as a white solid. ES-MS m/z 693 (MA-1).
Preparation 345
Methyl 3-fluoro-5-((1-methylpyrrolidin-3-yl)oxy)-4-nitrobenzoate
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F
0
02N
OMe
0
Stir a solution of methyl 3-fluoro-5-hydroxy-4-nitrobenzoate (2.0 g, 9.3
mmol), 1-
methylpyrrolidin-3-ol (1_13 g, 11_2 mmol), triphenylphosphine (2.93 g, 11.2
mmol), and
DIAD (2.26 g, 11.2 mmol) in THF (80 mL) at RT for 12 h. Filter and concentrate
the
reaction mixture and purify the residue via silica gel chromatography using a
gradient of
0 to 10% Me0H in DCM to give 1.03 g of the title compound (37%). ES-MS m/z 299
(M+H).
Preparation 346
Methyl 3-((1-methylpyrrolidin-3-yl)oxy)-4-nitro-5-((((S)-oxetan-2-
yl)methyl)amino)benzoate
%=== 0
H N
0
02N
0¨
0
Stir a solution of methyl 3-fluoro-5-(1-methylpyrrolidin-3-yl)oxy)-4-
nitrobenzoate
(1.03 g, 3.45 mmol), [(2S)-oxetan-2-yl]methanamine ( 20 g, 8.3 mmol), and TEA
(2.1 g,
21 mmol) in DMSO (150 mL) at 80 C for 12 h. Dilute the reaction with water
(50 mL)
and extract with Et0Ac (2 x 200 mL). Wash the combined organic layers with
saturated
aqueous NaC1 (5 x 40 mL). Dry the organic layer over MgSO4, filter, and
concentrate.
Purify the residue via silica gel chromatography using a gradient of 0 to 20%
Me0H in
DCM followed by purification via reversed phase chromatography using a
gradient of 30
to 60% MeCN in aqueous ammonium hydroxide (0.05%) to give 520 mg of the title
compound (41%) as a yellow oil. ES-MS m/z 366 (M+H).
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Preparation 347
Methyl 4-amino-3-((1-methylpyrrolidin-3-yl)oxy)-5-((((S)-oxetan-2-
yl)methyl)amino)benzoate
....
/ 0
HN
0
H2N
0-
0
tN,
Stir a solution of methyl 3-((1-methylpyrrolidin-3-yl)oxy)-4-nitro-5-((((S)-
oxetan-
2-yl)methyl)amino)benzoate (500 mg, 1.37 mmol) and palladium on carbon (10%,
200
mg, 0.19 mmol) in Et0Ac (100 mL) at RT under an atmosphere of hydrogen gas for
2 h.
Filter and concentrate the reaction mixture to give 358 mg of the title
compound (78%) as
a light yellow solid. ES-MS iniz 336 (M+H).
Preparation 348
Methyl 4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-34(1-methylpyrrolidin-3-yl)oxy)-5-
((((S)-
oxetan-2-y1)methyl)amino)benzoate
0
0 N 0 N 110 0
0
I ON.
To a solution of 2-(54-cy ano-3,9-dioxa-2(2,6)-py iidina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid (75 mg, 0.18 mmol) and HATU (100 mg,
0.26 mmol) in DMF (1.8 mL), add methyl 4-amino-3-((1-methylpyrrolidin-3-
yl)oxy)-5-
((((S)-oxetan-2-yl)methyl)amino)benzoate (67 mg, 0.20 mmol) and DIPEA (0.08
mL,
0.45 mmol). Stir at 20 'V for 2 h. Dilute the mixture with water (10 mL) and
extract with
Et0Ac (2 x 10 mL). Wash the combined organic layers with saturated aqueous
NaC1 (10
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mL), dry over Na9SO4, filter, and concentrate to give 425 mg of the title
compound,
which is used without further purification in Preparation 349. ES-MS in/z 718
(M+H).
Preparation 349
Methyl 2-454-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)methyl)-4-((1-methylpyrrolidin-3-y1)oxy)-1-(((S)-oxetan-2-y1)methyl)-1H-
benzo[d]imidazole-6-carboxylate
NC
NI 0
0 N
0-
0
Stir a solution of methyl 4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-3-((1-methylpyrrolidin-3-yl)oxy)-5-
((((S)-
oxetan-2-yl)methyl)amino)benzoate (Preparation 348, 425 mg, 0.528 mmol) in 1,2-
dichloroethane (4 mL) and acetic acid (4 mL) at 55 C for 12 h. Dilute with
1:1
toluene:ACN (10 mL) and concentrate the solution. Purify the residue via
silica gel
chromatography using a gradient of 0 to 10% Me0H in DCM to give 100 mg of the
title
compound (27%) as a white solid. ES-MS nilz 700 (M+H).
Preparation 350
Ethyl 2-(4-bromo-24(2-4(6-bromo-5-fluoropyridin-2-yl)oxy)methyl)-5-
cyanophenethoxy)methyl)-5-fluorophenyl)acetate
0
NC 0
Br
0 rN B
F
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To a solution of 4-[(6-bromo-5-fluoro-2-pyridyl)oxymethy11-3-(2-
hydroxyethyl)benzonitrile (4 g, 10.59 mmol), ethyl 244-bromo-2-(bromomethyl)-5-
fluoro-phenyl]acetate (8.3 g, 21 mmol) and 2,6-di-tert-butyl-4-methylpyri dine
(4.45 g,
21.2 mmol) in DCM (60 mL) add silver trifluoromethanesulfonate (8.3 g, 32
mmol) at 20
C. Stir the reaction at RT overnight, then concentrate the reaction mixture
under reduced
pressure to remove the solvent. Dilute the residue with water (100 mL) and
extract with
Et0Ac (100 mL x 2). Combine the organic layers, wash with saturated aqueous
NaCl (60
mL), dry over Na2SO4, filter and concentrate under reduced pressure. Purify
the residue
by silica gel chromatography using a gradient of 0% to 100% DCM in petroleum
ether to
give the title compound as a pale-yellow solid (2.15g, 25%) ES-MS miz 625
(M+H).
Preparation 351
Ethyl 2-(54-cyano-16,23-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetate
NC 0
0 N 0
To a mixture of ethyl 2-(4-bromo-2-02-4(6-bromo-5-fluoropyridin-2-
yl)oxy)methyl)-5-cyanophenethoxy)methyl)-5-fluorophenyl)acetate (2.15 g, 2.63
mmol)
and cesium fluoride (795 mg, 5.18 mmol,) in 1,4-dioxane (230 mL) under
nitrogen at RT,
add (2-di cyclohexylphosphino-2',4',6' -trii sopropyl- 1, l' -biphenyl) [2 -
(2'-amino-1, 1'-
biphenyl)]palladium(II) methanesulfonate (XPhos Pd G3, 450 mg, 0.521 mmol) and
hexamethylditin (1.34 g, 4.05 mmol). Protect the reaction from light heat the
reaction to
110 C and stir for 15 h, then concentrate the reaction mixture under reduced
pressure to
remove the solvent. Dilute the residue with water (100 mL) and extract with
Et0Ac (100
mL >< 3). Combine the organic layers, wash with saturated aqueous NaCl (60
mL), dry
over Na2SO4, filter and concentrate under reduced pressure. Purify the crude
residue by
preparative HPLC [column: Xtimate C18 150 x 40 mm, 10 m; mobile phase:
gradient of
50 to 90% ACN in water (with N1-13H20+NH4HCO3)] to give the title compound as
a
white solid. (360.1 mg, 29%) ES-MS nilz 465 (M+H).
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Preparation 352
2-(54-Cyano-16,23-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetic acid
N C 0
0 H
0 N 0
I F
Purge gently with nitrogen a solution of ethyl 2-(54-cyano-16,23-difluoro-3,8-
dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetate (345
mg,
0.74 mmol) in THF (2.47 mL), water (2.47 mL) and ACN (7.41 mL) and add 1,5,7-
triazabicyclo[4.4.0]dec-5-ene (320 mg, 2.25 mmol). Seal the reaction vessel
and purge
with nitrogen. Heat the reaction mixture to 45 C and stir at this temperature
for 2 h then
concentrate the reaction under reduced pressure to give the title compound as
a white
solid (385 mg, 100%) which is used in Preparation 353 without further
purification. ES-
MS nilz 437 (M+H).
Preparation 353
Methyl (S)-4-(2-(54-cyano-16,23-difluoro-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate
/1-0
N C 0 H N
O¨
N
0
0 N 0 0
I F
0
To a solution of 2-(54-cyano-16,23-difluoro-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetic acid (Preparation 352, 250
mg, 0.48
mmol) and HATU (378 mg, 0.97 mmol, ) in DMF (6 mL), add methyl 4-amino-3-(2-
methoxyethoxy)-5-[[(2S)-oxetan-2-yl]methylamino]benzoate (260 mg, 0.75 mmol),
then
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add DIPEA (300 pL, 1.72 mmol) to the mixture and stir at 20 C under N2
overnight.
Dilute the reaction with water (20 mL) and extract with Et0Ac (20 mL x 2).
Combine the
organic layers, wash with saturated aqueous NaC1 (20 mL), dry over Na2SO4,
filter and
concentrate under reduced pressure to give the titled compound as pale-yellow
solid (1.2
g, 100%) which is used in Preparation 354 without further purification ES-MS
m/z 729
(M+H).
Preparation 354
Methyl (S)-24(54-cyano-16,23-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate
NC 0
/"µ 0
N = 0¨
,
0
F
0
0
To a solution of methyl (S)-4-(2-(54-cyano-16,23-difluoro-3,8-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetamido)-3 -(2-
methoxyethoxy)-
5-((oxetan-2-ylmethyl)amino)benzoate (Preparation 353, 1.2 g, 0.49 mmol) in
1,2-
dichloroethane (5 mL) at RT under N2 add acetic acid (5 mL). Heat the reaction
at 55 C
and stir overnight. Dilute the reaction with toluene/ACN 1:1 (5 mL), remove
the solvent
under reduced pressure and purify the residue by silica gel chromatography
using a
gradient of 0 to 5% Me0H in DCM to give the title product as a white solid
(270 mg,
35%). ES-MS m/z 711 (M+H)
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Preparation 355
Methyl (S)-2-(chloromethyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
0
O¨
F
Cool a solution of methyl 4-amino-3-fluoro-5-[[(2S)-oxetan-2-
ylmethyl]aminoThenzoate (2.0 g, 7.9 mmol) and TEA (1.1 mL, 7.9 mmol) in 1,2-
dichloroethane (22 mL) to 0 C and add 2-chloroacetyl chloride (0.63 mL, 7.9
mL). Stir
the mixture at RT for 2 h. Dilute the reaction mixture with DCM and wash with
water.
Dry the organic layer over MgSO4, filter, and concentrate. Dissolve the
resulting residue
in acetic acid (40 mL) and stir at 70 C for 2 h. Concentrate the crude
reaction and purify
the residue via silica gel chromatography using a gradient of 0 to 4% Me0H in
DCM to
give 1.76 g of the title compound (72%) as a yellow oil. ES-MS m/z 313 (M-41).
Preparation 356
3-(2-((5-Bromo-2-chlorobenzyl)oxy)ethyl)-4-4(6-bromopyridin-2-
yl)oxy)methyl)benzonitrile
CI
NC 0
Br
0 N Br
Cool a solution of 4-[(6-bromo-2-pyridyl)oxymethy11-3-(2-
hydroxyethyl)benzonitrile (2.3 g, 6.8 mmol), 4-brom o-2-(bromomethyl)-1-chl
oro-
benzene (2.7 g, 9.5 mmol), and 2,6-di-tert-butylpyridine (2.2 mL, 9.7 mmol) in
DCM (36
mL) to 0 C. Add silver trifluoromethanesulfonate (3.0 g, 11.8 mmol) to the
reaction
mixture and stir at 0 C for 15 min before stirring at RT for 18 h. Add
additional silver
trifluoromethanesulfonate (0.53 g, 2.1 mmol) to the reaction mixture and stir
at RT for 22
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h. Filter the crude reaction mixture through Celitec), concentrate, and purify
the residue
via silica gel chromatography using a gradient of 0 to 100% DCM in heptane to
give 2.3 g
of the title compound (62%) as a colorless sticky solid. ES-MS nilz
535/537/539 (M+H).
Preparation 357
14-Chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-54-
carbonitrile
NC 0
CI
0 N
I
Prepare the title compound essentially as described in Preparation 216 using 3-
(2-
((5-bromo-2-chl orobenzyl)oxy)ethyl)-4-(((6 -bromopyri di n-2-yl)oxy)methyl
)benzonitrile,
purify the title compound via silica gel chromatography using a gradient of 0
to 25%
Et0Ac in heptane to give the title compound as beige solid. ES-MS m/z 377
(M+H).
Preparation 358
14-(4,4,5,5-Tetramethy1-1,3,2-dioxaborolan-2-y1)-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-54-carbonitrile
NC 0
B-0
0 N
I
Stir a solution of 14-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-54-carbonitrile (170 mg, 0.45 mmol), (S)-1,2-
propanediol (0.2
mL, 3 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (XPhos, 44
mg, 0.09
mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1, 1 '-bipheny1)[2-
(2'-amino-
1,1'-biphenyl)]palladium(II) (XPhos Pd Gen 2, 35 mg, 0.045 mmol),
bis(pinacolato)diboron (0.34 g, 1.3 mmol), and KOAc (0.13 g, 1.3 mmol), in 2-
methyltetrahydrofuran (14 mL) at 80 C; for 3 h. Filter and concentrate the
reaction
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solution. Dissolve the residue in Et0Ac and wash with water. Dry the organic
layer over
MgSO4, filter, and concentrate to give 210 mg of the title compound (99%),
which is used
in Preparation 359 without further purification. ES-MS nilz 469 (M-41).
Preparation 359
Methyl (S)-2-054-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
N
0
1 0
0 N
1 0¨
/
Stir a solution of 14-(4,4,5,5-tetram ethyl -1,3,2-di oxaborol an-2-y1)-3,8-di
oxa-
2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-54-carbonitrile (0.12 g,
0.26
mmol), methyl (S)-2-(chloromethyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate (0.12 g, 0.38 mmol), 2-dicyclohexylphosphino-
2',4',6'-
triisopropylbiphenyl (XPhos, 25 mg, 0.05 mmol), chloro(2-dicyclohexylphosphino-
2',4',6'-triisopropy1-1, 1'-bipheny1)[2-(21-amino-1,1'-biphenyl)]palladium(II)
(XPhos Pd
Gen 2 20 mg, 0.025 mmol), and potassium phosphate tribasic (67 mg, 0.31 mmoL)
in 2-
methyltetrahydrofuran (3 mL) and water (0.3 mL) at 90 C for 7 h. Filter the
reaction
solution and dilute with Et0Ac, wash with water, dry the organic layer over
MgSO4,
filter, and concentrate. Purify the residue via silica gel chromatography
using a gradient
of 0 to 50% Et0Ac in DCM to give 20 mg of the title compound (10%) as a yellow
solid.
ES-MS nilz 619 (M-F1-1).
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Preparation 360
Methyl (S)-3-(2-(dimethylamino)-2-oxoethoxy)-4-nitro-5-((oxetan-2-
ylmethyl)amino)benzoate
0
HN
02N
0
Add sodium hydride in mineral oil (60 wt%, 211 mg, 5.28 mmol) to a solution of
2-hydroxy-N,N-dimethylacetamide (730 mg, 708 mmol) in TUE (10 mL) at 0 C,
then
remove the cooling bath and stir the mixture for 1 h. Re-cool the mixture to 0
C and add
methyl 3-fluoro-4-nitro-5-[[(25)-oxetan-2-ylmethyl]amino]benzoate (1 g, 3.52
mmol).
Remove the cooling bath and stir the mixture for 48 h. Quench the reaction
with
saturated NH4C1 solution (30 mL) and extract with Et0Ac (3 x 50 mL). Separate
the
organic layer, dry over Na2SO4, filter and concentrate. Purify the residue by
silica gel
chromatography using a gradient of 0 to 60% Et0Ac in petroleum ether to afford
the title
compound as a yellow oil (800 mg, 61%). ES-MS nilz 368 (M+H).
Preparation 361
Methyl (S)-4-amino-3-(2-(dimethylamino)-2-oxoethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate
0
0
HN
H 2 N
0
N0
Purge a vessel containing methyl (S)-3-(2-(dimethylamino)-2-oxoethoxy)-4-nitro-
5-((oxetan-2-ylmethyl)amino)benzoate (800 mg, 2.13 mmol), Pd/C (200 mg, 10
wt%),
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and Et0Ac (60 mL) with hydrogen gas three times. Stir the mixture under one
atmosphere of H2 for 4 h at ambient temperature. Filter the mixture and
concentrate
under reduced pressure. Purify the product by silica gel chromatography using
a gradient
of 0 to 100% Et0Ac in petroleum ether to afford the title compound as a yellow
oil
(636.4 mg, 88%). ES-MS m/z 338 (M+H).
Preparation 362
Methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-(dimethylamino)-2-oxoethoxy)-5-
((oxetan-2-ylmethyl)amino)benzoate
N H N 000
N
0¨
0 N 0 c! 0
,
N¨
/
At ambient temperature, add 1-propanephosphonic anhydride (50 wt% solution in
DCM, 225 FL, 0.378 mmol) to a solution of 2-(54-cyano-3,9-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetic acid (75 mg, 0.187 mmol)
and
methyl (S)-4-amino-3-(2-(dimethylamino)-2-oxoethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate (69 mg, 0.20 mmol) in DMF (1 mL) and pyridine (150 uL,
1.85
mmol). Stir for 20 h and then add additional 1-propanephosphonic anhydride
(225 uL,
0.378 mmol). After stirring another 23 h, quench the reaction by adding water
(2 mL).
Stir the mixture for 10 min, filter and collect the solid, then dry the solid
at 60 C under
vacuum for 2 h to give the title compound as a beige solid (75.1 mg, 56%). ES-
MS m/z
720 (M+H).
Preparation 363
Methyl (S)-2-054-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-(dimethylamino)-2-oxoethoxy)-1-
(oxetan-
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2-ylmethyl)-1H-benzoldlimidazole-6-carboxylate
NC
0
N
0
0 N N
0
0
/ 0
Stir a solution of methyl (S)-4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)acetamido)-3-(2-(dimethylamino)-2-
oxoethoxy)-5-((oxetan-2-ylmethyl)amino)benzoate (75 mg, 0.104 mmol) in acetic
acid (3
mL) at 60 C for 20 h. Remove the acetic acid under vacuum and purify the
residue via
silica gel chromatography using a gradient of 0 to 20% isopropanol in
chloroform to give
70 mg of the title compound (70% purity, 72%). ES-MS nilz 703 (M+H).
Preparation 364
Methyl 343-ethylimidazol-4-yemethylamino]-5-methoxy-4-nitro-benzoate
/7--N
0
HN
0
02N
o
Add TEA (2.2 mL, 16 mmol) to a stirred solution of methyl 3-fluoro-5-methoxy-
4-nitro-benzoate (900 mg, 3.93 mmol) and (1-ethyl-1H-imidazol-5-yl)methylamine
dihydrochloride (900 mg, 4.32 mmol) in DMF (20 mL). Stir the mixture at 60 C
for 16
h. Allow the mixture to cool to ambient temperature and pour into saturated
aqueous
NH4C1 (100 mL). Extract the mixture with DCM (3 x 100 mL) and wash with
saturated
aqueous NaCl (3 x 50 mL). Dry the organics over Na2SO4, filter, and
concentrate under
reduced pressure. Purify the residue by flash chromatography using a gradient
of 0 to
10% Me0H in Et0Ac to give the title compound (800 mg, 61%) as a yellow oil. ES-
MS
nilz 335 (M+H).
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Preparation 365
Methyl 4-amino-3-(((1-ethyl-1H-imidazol-5-yl)methyl)amino)-5-methoxybenzoate
0
H 2N
0
Add iron (801 mg, 14.34 mmol) and NE4C1 (768 mg, 14.4 mmol) to a solution of
methyl 3-1(3-ethylimidazo1-4-yl)methylamino]-5-methoxy-4-nitro-benzoate (960
mg,
2.87 mmol) in Me0H (30 mL) and water (10 mL) Stir at 80 C for 2 h, cool to
ambient
temperature, then filter over a pad of Celite and wash the pad with Me0H (30
mL).
Concentrate the filtrate under reduced pressure to and purify by flash
chromatography
using a gradient of 0 to 15% Me0H in Et0Ac. Re-purify by silica gel
chromatography
using a gradient of 0 to 10% Me0H in Et0Ac to give the title compound as a
brown solid
(504 mg, 55%). ES-MS nilz 305 (M+H).
Preparation 366
Methyl 4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-ypacetamido)-34((1-ethyl-1H-imidazol-5-
yl)methyl)amino)-5-methoxybenzoate
N
H N
0
0
0 N 0
, 0
At ambient temperature, add 1-propanephosphonic anhydride (50 wt% solution in
DMF, 170 pL, 0.28 mmol) to a solution of 2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetic acid (76 mg, 0.1898 mmol)
and
methyl 4-amino-3-(((1-ethyl-1H-imidazol-5-yl)methyl)amino)-5-methoxybenzoate
(63
mg, 0.207 mmol) in DMF (600 pL) and pyridine (400 pL, 5 mmol). Stir the
reaction at
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RT for 72 h, then dilute with Et0Ac (100 mL), wash the organics with saturated
NaHCO3
(100 mL), filter over Celite, and extract the filtrate three times with
additional Et0Ac.
Combined the organics, wash twice with saturated aqueous NaC1, dry over MgSO4,
filter,
and concentrate under reduced pressure to afford the title product (136 mg)
which is used
in Preparation 367 without further purification ES-MS m/z 688 (M+H).
Preparation 367
Methyl 24(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)methyl)-141-ethyl4H-imidazol-5-y1)methyl)-4-methoxy-1H-benzo[d]imidazole-
6-
carboxylate
NNj
0
I / 0
0 N
,
0-
0
Prepare the title compound essentially as described in Preparation 102 using
methyl 4-(2-(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)acetamido)-3-(((1-ethyl-1H-imidazol-5-yl)methyl)amino)-5-methoxybenzoate
(Preparation 366) in a mixture of 1:1 acetic acid: 1,2-dichloroethane as
solvent, heating
the reaction at 70 C for 16 h. Cool the mixture to RT and concentrate under
vacuum.
Add 1:1 Et0Ac/toluene and concentrate to remove residual acetic acid (twice).
Dry the
resulting yellow solid under vacuum for 2 h. Purify the solid by silica gel
chromatography
using a gradient of 90 to 100% Et0Ac in DCM, followed by Me0H, to give the
title
compound as an off-white solid which is used in Example 63 without further
purification.
ES-MS m/z 670 (M+H).
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Preparation 368
Methyl (S)-4-(2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)acetamido)-3-(2-methoxyethoxy)-5-((oxetan-2-
ylmethyl)amino)benzoate
NC 0 H
0 N 0 o 0
0
To a solution of 2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-y1) acetic acid (44 mg, 0.103 mmol) in DMF (1.2
mL), add
methyl 4-amino-3-(2-methoxyethoxy)-5-[[(2S)-oxetan-2-yl]methylamino]benzoate
(32
mg, 0.103 mmol), HATU (59 mg, 0.155 mmol) and DIPEA (0.055 mL, 0.32 mmol).
Stir
the mixture at RT for 2 h, then dilute with water (10 mL) and extract with
Et0Ac (4x 5
mL). Combine the organics, wash with water and saturated aqueous NaCl, dry
over
MgSO4, filter, and concentrate under vacuum to give the title compound as a
light brown
solid (80 mg, 80% purity, 88% yield). ES-MS nilz 693 (M+H).
Preparation 369
Methyl (S)-2-054-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate
NC 0
/ 0
0 N
I 0-
0
0
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Heat a solution of methyl (S)-4-(2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-ypacetamido)-3-(2-methoxyethoxy)-5-
((oxetan-2-ylmethyl)amino)benzoate (80 mg, 80% purity, 0.092 mmol) in 1,2-
dichloroethane (0.7 mL) and acetic acid (0.7 mL) at 58 C for 6 h. Cool
reaction mixture
to RT, concentrate under reduce pressure, and purify the residue via silica
gel
chromatography using using a gradient of 30 to 100% Et0Ac in DCM to provide 25
mg
(40%) of the title compound as a white solid. ES-MS m/z 675 (M+H).
Preparation 370
Methyl (S)-24(54-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-
14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylate
I I
0
0 N 0
0--
0
0
To a solution of 2-(54-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)acetic acid (60 mg, 0.15 mmol) and methyl 4-
amino-3-
(2-methoxyethoxy)-5-[[(2S)-oxetan-2-yl]methylaminoThenzoate (47 mg, 0.15) in
anhydrous DMF (1.5 mL) under nitrogen atmosphere, add HATU (74 mg, 0.19 mmol)
and DIPEA (0.08 mL, 0.45 mmol). Stir the mixture at RT for 2.5 h, then add
water and
Et0Ac. Separate the aqueous layer and wash the organic layer with water and
saturated
aqueous NaCl, dry over Na2SO4, filter and concentrate. Heat a solution of the
residue in
1,2-dichloroethane (0.9 mL) and acetic acid (0.9 mL) under nitrogen atmosphere
at 60 'V
for 8 h. Cool reaction mixture to RT, concentrate the solvents under reduced
pressure, dry
under vacuum at 35-40 C and purify the residue via silica gel chromatography
using a
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gradient of 25 to 100% of Et0Ac in DCM as eluent system to provide the title
compound
(72 mg, 71%) as a white solid_ ES-MS nilz 675 (M+H).
Preparation 371
(S)-(1446-(Methoxycarbony1)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazol-2-
y1)methyl)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-54-
y1)boronic acid
0 H
HOBi
' 0
0 N 0
0
Charge a reaction vessel with methyl (S)-2454-bromo-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate (500 mg, 0.72 mmol), anhydrous THF (5 mL) and
Me0H (10 mL). Bubble the mixture with N2 for 10 min, add anhydrous ethylene
glycol
(610 ttL, 10.9 mmol) and DEPEA (315 ulõ 1.82 mmol). Bubble the mixture with N2
for 5
min, add tetrahydroxydiboron (139 mg, 1.47 mmol), tricyclohexylphosphine (5
mg, 0.018
mmol) and [(tricyclohexylphosphine)-2-(2'-aminobiphenyl)]palladium(H)
methanesulfonate [P(Cy3) Pd G3, 26 mg, 0.039 mmol), seal the vessel and stir
at 50 C in
a preheated bath for 2.5h. Concentrate the reaction mixture, then add
saturated aqueous
NaHCO3 and stir for 5 min. Filter off the solid, then wash the solid with
water, ACN and
Me0H to provide the title compound (500 mg, 90 wt% pure, 100%) as a gray
solid. ES-
MS nilz 620 (M+H).
Preparation 372
Methyl (S)-2-((54-(4-fluoro-1H-imidazol-1-y1)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate
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N
=
0
0 N N
,
0
Charge a reaction vessel with (S)-(14-46-(methoxycarbony1)-1-(oxetan-2-
ylmethyl)-1H-
benzord]imidazol-2-yl)methyl)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-54-y1)boronic acid (21 mg, 0.03 mmol, 90 wt% pure), 4-
fluoro-1H-imidazole (14 mg, 0.15 mmol), copper(II) acetate (5.8 mg, 0.032
mmol),
Me0H (245 1..iL) and pyridine (6 [IL, 0.07 mmol). Seal the reaction vessel and
stir the
suspension at 60 C for 10 h. Add Et0Ac and aqueous ammonia (28%), separate
the
organic layer and wash with aqueous ammonia (28%), water and saturated aqueous
NaCl,
dry over Na2SO4, filter and concentrate. Purify the residue via silica gel
chromatography
using a gradient of 20 to 100% of Et0Ac in DCM as eluent system to provide the
title
compound (8 mg, 10%) as a white solid. ES-MS m/z 660 (M-41).
Preparation 373
Methyl 2-(bromomethyl)-5-chlorobenzoate
0
0
Br
CI
Prepare the title compound essentially as described in Preparation 236 using
methyl 5-chloro-2-methylbenzoate. Stir the reactor output with water and
aqueous
sodium bisulfite, separate the layers, then extract the aqueous phase twice
with heptane.
Combine the organics, wash with water (3 A), saturated aqueous NaHCO3 and then
saturated aqueous NaCl. Dry the organics over MgSO4, filter and concentrate to
give the
title compound (67.99 g, 87%) as a yellow oil. 1H NMR_ (400.21 MHz, CDC13) 6
7.98 (d,
J= 2.2 Hz, 1H), 7.49 (dd, J= 2.3, 8.3 Hz, 1H), 7.43 (d, J= 8.3 Hz, 1H), 4.94
(s, 2H), 3.97
(s, 3H).
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Preparation 374
Methyl 5-chloro-2-[(6-chloro-2-pyridyl)oxymethylibenzoate
0
CI
0
0 N CI
Add a solution of methyl 2-(bromomethyl)-5-chloro-benzoate (20.0 g, 67.5 mmol)
in toluene (200 mL) to a mixture of 6-chloropyridin-2-ol (10.9 g, 84.1 mmol)
and silver
carbonate (14.9 g, 54.0 mmol). Heat the mixture under N2 to 65 C for 48 h,
protecting
the reaction vessel from light using aluminum foil and adding additional
toluene (100
mL). Add DCM (200 mL) to the reaction and filter through a pad of Celiteg,
rinse the
pad with DCM (100 mL). Concentrate the filtrate to a volume of 100 mL, filter
off a first
crop of solid material. Wash the solid with 1:1 toluene/heptane (50 mL) and
heptane (2 X
50 mL). Add 100 mL heptane to the filtrate and then filter off a second crop
of solid
material, wash as before with 1:1 toluene/heptane (50 mL) and heptane (2 x 50
mL).
Concentrate the filtrate and slurry the residue in heptane (200 mL) at 50 C
for 30 min,
then stir at ambient temperature overnight. Filter off a third crop of solid
and wash the
solid with heptane (2 x 50 mL). Combine the first, second, and third crops of
solid
materials and dry under reduced pressure at 50 C for 5.5 h to give the title
compound
(18.85 g, 89%) as a white solid. ES/MS nilz 312, 314 (M+H).
Preparation 375
[5-Chloro-2-[(6-chloro-2-pyridyl)oxymethyl]phenyl]methanol
CI
Si OH
0 N CI
To a mixture of methyl 5-chloro-2-[(6-chloro-2-pyridyl)oxymethylibenzoate
(14.78 g, 46.87 mmol) in TI-IF (75 mL) under N2 add LiBH4 (2 M in THF, 35 mL,
70
mmol). Stir the reaction mixture for 5 min at ambient temperature, then add
Me0H (2.9
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mL, 72 mmol) portionwise over 1 h. Add Et0Ac (5 mL), water (10 mL), HC1 (1 M
aqueous, 100 mL), and MTBE (300 mL) to the mixture and separate the layers.
Separate
the layers, wash the organics with water (50 mL), aqueous K2CO3 (2 M, 50 mL),
and
saturated aqueous NaC1 (50 mL) Dry the organics over MgSO4, filter and
concentrate
the filtrate to give the title compound (13.54 g, 97%) as a waxy solid. ES/MS
m/z 284,
286 (M+H).
Preparation 376
24[245-Bromo-4-fluoro-2-iodo-phenyl)methoxymethyl]-4-chloro-phenyl]methoxy]-6-
chloro-pyridine
Br
14111
CI 0
0 N CI
Add THF (7.5 mL) to a mixture of [5-chloro-2-[(6-chloro-2-
pyridyl)oxymethyl]phenyl]methanol (0.50 g, 1.7 mmol) and 1-bromo-5-
(bromomethyl)-
2-fluoro-4-iodobenzene (0.89 g, 2.1 mmol) under N2, then add potassium tert-
butoxide (1
M solution in tert-butanol, 2.2 mL, 2.2 mmol) portionwi se. Stir the mixture
at RT for 30
min, then add water (30 mL) and stir the mixture at RT overnight, resulting in
a mixture
with a sticky lower phase. Decant off the supernatant, add water and decant
off the water.
Dissolve the remainder in Me0H (55 mL) with heating at 60 C and add SiliaMetS
Triamine (1 g) and continue heating at 60 C for 3.5 h. Filter the reaction
while hot
through a pad of Celite , rinse the pad with hot Me0H (15 mL), and concentrate
the
filtrate. Dissolve the residue in MTBE (20 mL), filter, and concentrate.
Purify the reside
by silica gel chromatography using a gradient of 0 to 20% Et0Ac in cyclohexane
to give
the title compound (0.75 g, 68%) as a colorless oil. ES/MS rn/z 595, 597,
599(M+H).
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Preparation 377
Ethyl 2-[4-bromo-2-[[5-chloro-2-[(6-chloro-2-
pyridyl)oxymethyl]phenyl]methoxymethy11-5-fluoro-phenyl]acetate
Br 0
CI
0
0 N CI
Under N2, add bromo-(2-ethoxy-2-oxo-ethyl)zinc (0.4 M in THF, 3.4 mL, 1.4
mmol) to a mixture of 24[245-bromo-4-fluoro-2-iodo-pheny1)methoxymethy1]-4-
chloro-phenyl]methoxy]-6-chloro-pyridine (0.58 g, 0.91 mmol) and chloro[(4,5-
bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2'-amino-1,1'-
biphenyl)]palladium(II)
(Pd-179, Xantphos Pd G2, 45 mg, 0.045 mmol) in THF (1 mL). Heat the reaction
mixture to 60 C for 10 h. Partition the reaction mixture between water (15
mL), aqueous
citric acid (5%, 5 mL), and MTBE. Wash the organic phase with 5 mL portions of
water,
aqueous K2CO3 (2 M), and saturated aqueous NaCl. Concentrate the organics onto
Celite and purify by silica gel chromatography using a gradient of 5 to 40%
Et0Ac in
cyclohexane to give the title compound (311 mg, 58%) as a colorless oil. 1H
NMR
(400.13 MHz, CDC13) 6 7.56-7.52 (m, 2H), 7.46-7.44(m, 2H), 7.32 (dd, J= 2.2,
8.1 Hz,
1H), 7.07 (d, J= 9.0 Hz, 1H), 6.94 (d, J= 6.8 Hz, 1H), 6.66 (d, J= 7.6 Hz,
1H), 5.36 (s,
2H), 4.66 (s, 2H), 4.57 (s, 2H), 4.11 (q, J= 7.1 Hz, 2H), 3.68 (s, 2H), 1.22
(t, J= 7.1 Hz,
3H).
Preparation 378
Ethyl 2-(54-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yDacetate
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CI =0,-
0 N 0
F
Sparge N2 for 10 min into a mixture of ethyl 2-[4-bromo-21[5-chloro-2-[(6-
chloro-2-pyridyl)oxymethyl]phenyl]methoxymethy1]-5-fluoro-phenyl]acetate (0.59
g, 1.0
mmol), bis(neopentyl glycolato)diboron (0.28 g, 1.2 mmol) and potassium
pivalate (0.36
g, 2.5 mmol) in anhydrous THE' (40 mL), then add chloro(2-
dicyclohexylphosphino-
2',4',6'-tri-i-propy1-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-y1)
palladium(II) (X-Phos-Pd-
G2, 42 mg, 0.052 mmol). Heat the reaction mixture at 45 C for 1.5 h, then 55
C for 1 h
then add more bis(neopentyl glycolato)diboron (46 mg, 0.20 mmol) and continue
heating
at 55 C for 45 min. Add potassium phosphate tribasic (1.0 M solution in
water, 3 mL,
3.0 mmol) and continue heating at 55 C for 2 h. Partition the reaction
mixture between
aqueous K9CO3 (2 M, 25 mL) and DCM (100 mL) and separate the layers. Extract
the
aqueous layer with DCM (25 mL), combine the organics and filter through Ce
Concentrate the filtrate and purify the residue by silica gel chromatography
using DCM.
Triturate the product with a mixture of DCM (5 mL) and heptane (20 mL) and dry
the
solid under vacuum at 40 C to give the title compound (144 mg, 32%) as a
white solid.
ES/MS raiz 442, 444 (M+H).
Preparation 379
2-(54-Chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-
14-yOacetic acid
CI 010 0
0 H
0 N 0
F
Add LiOH (1M aqueous, 2.1 mL, 2.1 mmol) to a suspension of ethyl 2-(54-chloro-
16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclooctaphane-14-
yl)acetate
(227 mg, 0.51 mmol) in a mixture of THF (7 mL) and Me0H (3.4 mL). Heat the
mixture
at 60 "V for 1 h. Concentrate the reaction mixture and add aqueous citric acid
(5%).
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Filter off the solid, wash it with water and dry it under vacuum at 40 C to
obtain the title
compound (246 mg, 90 mass%, 100%) as a white solid. ES-MS rn/z 414, 416 (M+H).
Preparation 380
Methyl (S)-2-054-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate
CI
4111 0 =
NI?
0 N N 0
0
F
0¨/¨O
To a solution of 2-(54-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)acetic acid (246 mg, 0.535 mmol, 90 mass%) and
methyl
4-amino-3-(2-methoxyethoxy)-5-[[(2S)-oxetan-2-yl]methylaminoThenzoate (183 mg,
0.59
mmol) in anhydrous DMF (6 mL) under nitrogen atmosphere, add pyridine (492 uL,
6.08
mmol) and 2,4,6-tripropy1-1,3,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxide
(1.68 M in
EtoAc, 800 uL, 1.34 mmol). Stir the mixture at RT for 30 min, then add water.
Filter off
the solid, wash with water, and dry at 40 C overnight. Heat a suspension of
the solid in
1,2-dichloroethane (6.4 mL) and acetic acid (6.4 mL) under N2 atmosphere at 60
C
overnight. Cool the reaction mixture, dilute with Et0Ac and water and filter
off the solid.
Separate the organic layer, dry over Na2SO4, filter and concentrate the
organics under
reduce pressure. Purify the residue via silica gel chromatography using a
gradient of 0 to
50% Et0Ac in DCM to provide the title compound (291 mg, 75%) as a white solid.
ES-
MS m/z 688, 690 (M+H).
Example 1
(S)-2-((54-Cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
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N C
0 NN CO2 H
F
Stir a mixture of methyl (S)-2454-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-
benzord]imidazole-6-carboxylate, (200 mg, 0.257 mmol) and 1,5,7-
triazabicyclo[4.4.0]dec-5-ene (300 mg, 2.11 mmol) in ACN:water (4.0 mL:1.0 mL)
at 60
C for 5 h. Adjust the mixture to pH 6 with 1.0 M aqueous hydrochloric acid
solution.
Purify the reaction mixture in its entirety via C18 reversed phase
chromatography eluting
with a gradient of 40 to 70% ACN in 0.225% aqueous formic acid to give 33 mg
of the
title compound (21%). ES-MS m/z 605 (M+H).
Example 2
(S)-24(54-Cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid
N C . , 0
0 N CO2 H
I F
To a solution of methyl (S)-2454-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-4-fluoro-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate (0.24 g, 0.38 mmol) in ACN:water (5.0 mL:
1.0 mL)
add 1,3,4,6,7,8-hexahydro-2h-pyrimido[1,2-a]pyrimidine (50 mg, 0.40 mmol).
Stir at RT
for 15 h then at 60 C for 4 h. Concentrate the reaction mixture to half volume
and
neutralize to pH 7 with 1 M aqueous citric acid solution. Dilute the mixture
with water
(100 mL) and extract with DCM (3 50 mL). Wash the combined organic layers with
saturated aqueous sodium chloride (50 mL). Dry the organic phase over sodium
sulfate,
filter, and concentrate under reduced pressure. Purify the residue via silica
gel
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chromatography using a gradient of 5 to 100% solvent B in solvent A, where
solvent B is
20% Me0H in Et0Ac and solvent B is DCM. Further purify the product via C18
reversed phase chromatography using a gradient of 25 to 40% ACN in 10 mM
aqueous
ammonium bicarbonate containing 5% Me0H to give 40 mg of the title compound
(17%). ES-MS nilz 623 (M+H).
Example 3
(S)-24(54-Chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
CI 0
0 N N = CO2 H
To a mixture of methyl (S)-2454-chloro-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate (45 mg, 0.074 mmol) in ACN:THF:Me0H (0.80 mL:0.50 mL:0.50 mL) add
aqueous lithium hydroxide solution (1.0 M, 0.75 mL). Stir the mixture at 40 C
for 6 h
and 55 C for 30 min. Adsorb the mixture onto Celite and purify via C18
reversed phase
chromatography eluting with a gradient of 0 to 100% ACN in 10 mM aqueous
ammonium bicarbonate with 5% Me0H to give 22 mg of the title compound (49%).
ES-
MS rn/z 596 (M+H).
Example 4
(S)-2-((54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
NC
0
0 N N = CO21-1
I
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Stir a mixture of methyl (S)-24(54-cyano-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate, (240 mg, 0.40 mmol) and 1,5,7-triazabicyclo[4.4.0]dec-5-ene (170
mg, 1.20
mmol) in 1,4-dioxane : ACN: water (5:5:1, 11 mL) at 60 C for 3h, 25 C for 16
h, then
50 C for 72 h. Concentrate the mixture to one-quarter volume and purify via
C18
reversed phase chromatography eluting with a gradient for 10 to 80% ACN in 10
mM
aqueous ammonium bicarbonate containing 5% Me0H to give 160 mg of the title
compound (68%). ES-MS m/z 587 (M+H).
Example 5
(S)-2-((54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yOmethyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic
acid
NC
Cs?
O)1)) N CO2H
0
Stir a mixture of methyl (S)-24(54-cyano-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate, (230 mg, 0.28 mmol) and 1,5,7-
triazabicyclo[4.4.0]dec-5-ene (170 mg, 1.20 mmol) in 1,4-dioxane : ACN : water
(5 : 5 :
1, 11 mL) at 50 C for 16 h, 65 C for 4 h, then 50 C for 72 h. Concentrate
the mixture
to one-quarter volume and purify via C18 reversed-phase chromatography eluting
with a
gradient of 10 to 80% ACN in 10 mM aqueous ammonium bicarbonate containing 5%
Me0H to give the title compound (170 mg, 72%). ES-MS ni/z 617 (M+H).
Example 6
(S)-2-((54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-3-(oxetan-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylic acid
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NC
=
0
I N
0 N N )- CO2 H
Prepare the title compound essentially as described in Example 5 using methyl
(S)-2-((54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-3-(oxetan-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylate. ES-
MS 111/ Z 588 (M+H).
Example 7
(S)-2-454-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic
acid
NC
0
i""= 0
0 N = CO2H
I
Prepare the title compound essentially as described in Example 5 using methyl
(S)- 2 -((54-cyano-3 ,9-di oxa-2(2,6)-pyri di na-1 (1, 3 ), 5 (1 , 2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.
ES-
MS iniz 605 (M+H).
Example 8
(S)-2-054-Cyano-16-fluoro-9-oxa-3-aza-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1 -(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
NC
0 ". 0
I /
H NI1.1J N CO 2H
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Prepare the title compound essentially as described in Example 5 using methyl
(S)-2-((54-cyano-16-fluoro-9-oxa-3-aza-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-1 4-y1 )methyl)-1 -(oxetan -2-ylm ethyl)-11-1-
benzo[d]imidazol e-6-
carboxylate in ACN : TUT' : water (1: 1 : 0.4). Heat the mixture at 50 C for
4 h, cool to
RT and quench with 1M citric acid solution. Extract the mixture three times
with Et0Ac.
Combine the organics, wash with water and brine, dry over magnesium sulfate,
filter, and
concentrate under reduced pressure. Purify the residue by C18 reverse phase
chromatography using a gradient of 35 to 70% ACN in 20 mM aqueous ammonium
bicarbonate to give the title compound as a white solid. ES-MS m/z 604 (M+H).
Example 9
(S)-2454-Cyano-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid
NC
0 t 0
iThF
0 N N CO2H
0
Prepare the title compound essentially as described in Example 5 using methyl
(S)-2454-cyano-16-methy1-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dib enzenacy cl ononaphane- 1 4-yl)methyl) -4-methoxy - 1 -(oxetan-2-ylm
ethyl)- 1H-
b enzo[d]imidazole-6-carb oxylate. ES-MS nilz 631 (M+H).
Example 10
(S)-2-((54-Cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
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CN
0 CiO
0 N
N =
CO2H
Prepare the title compound essentially as described in Example 5 using methyl
(S)-24(54-cyano-16-fluoro-3,9-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-
14-yl)m ethyl )-1-(oxetan-2-ylmethyl)-1H-benzo[d]imi dazole-6-carboxyl ate.
Heat the
mixture at 60 C under nitrogen atmosphere for 2 h, then cool to RT and quench
with
citric acid (5% aqueous). Filter the solid, then wash with water and ACN to
give the title
compound as a white solid. ES-MS rn/z 605 (M+H).
Example 11
2-054-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)methyl)-1-((1-ethyl-1H-imidazol-5-y1)methyl)-1H-benzo[d]imidazole-6-
carboxylic
acid
NC
0
0 N N /11, CO2H
I
To a solution of methyl 2-454-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-((l-ethyl-1H-imidazol-5-yOmethyl)-1H-
benzo[d]imidazole-6-carboxylate (39.7 mg, 0.06 mmol) in ACN (0.75 mL), THF
(0.19
mL), and water (0.12 mL) add 1,3,4,6,7,8-hexahydro-2h-pyrimido[1,2-
a]pyrimidine (35
mg, 0.25 mmol). Stir the mixture at 45 C for 3 h. Add additional 1,3,4,6,7,8-
hexahydro-
2h-pyrimido[1,2-a]pyrimidine (7.5 mg, 0.05 mmol) and stir the reaction at 50
C for 1 h.
Quench the reaction to pH 6-7 with formic acid and extract with Et0Ac. Dilute
with
water and extract with Et0Ac. Dry the organic phase over magnesium sulfate,
filter, and
concentrate under reduced pressure. Purify the residue by C18 reversed phase
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chromatography using a gradient of 28 to 64% (1:1 ACN:Me0H) in (65 mM aqueous
ammonium acetate:ACN 90:10 solution) to give 11.8 mg of the title compound
(30%).
ES-MS rn/z 625 (M+H).
Example 12
(S)-14-((4-Methoxy-1-(oxetan-2-ylmethyl)-6-(1H-tetrazol-5-y1)- 1H-
benzo[d]imidazol-2-
yl)methyl)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-54-
carbonitrile
NC
0 / 0
0 N Ni 411, /NI
I N-N
Me0
Stir ( S)-2-(54-cyano-3, 9-di oxa-2(2,6)-pyri di na-1(1,3), 5(1,2)-
dibenzenacyclononaphane-14-y1)-N-(2-methoxy-6-((oxetan-2-ylmethyl)amino)-4-(1H-
tetrazol-5-yl)phenypacetamide (56 mg, 0.05 mmol) in acetic acid (1.0 mL) at 65
C for
12 h. Concentrate the solution and azeotrope with ACN. Purify the residue by
C18
reversed phase chromatography using a gradient of 41 to 83% 1:1 ACN:Me0H in 25
mM
aqueous ammonium carbonate solution to give 9.4 mg of the title compound
(28%). ES-
MS 117/Z 641 (M+H).
Example 13
(S)-2-((54-Cyano-16-fluoro-3,6,9-trioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
NC t 0
0 N CO2H
F
Prepare the title compound essentially as described in Example 1 using methyl
(S)-2-((54-cyano-16-fluoro-3,6,9-trioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
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dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzokilimidazole-6-
carboxylate, stirring the reaction at 45 C for 2 h. When complete, quench the
reaction
with formic acid to pH 7 and dilute the crude mixture with water. Extract the
mixture
three times with Et0Ac. Dry the organic phase over magnesium sulfate, filter,
and
concentrate under reduced pressure. Purify the residue via silica gel flash
chromatography
using a gradient of 0 to 40% 9:1 DCM:Me0H containing 1% formic acid in DCM to
give
the title compound. ES-MS iii/z 607 (M+H).
Example 14
(S)-2-((54-Cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
NC
0
OJtrJN = CO2H
F
Prepare the title compound essentially as described for Example 2 using methyl
(S)-2-((54-cyano-16-fluoro-3,7-di oxa-2(2,6)-pyri di n a -1 (1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate, stirring the reaction at 45 C for 19 h, then add 1,4-dioxane and
stir at 45 C
for 23 h. Quench the reaction to pH 6-7 with formic acid and extract with
Et0Ac,
followed by 3:1 chloroform:2-propanol. Dry the organic phase over magnesium
sulfate,
filter, and concentrate under reduced pressure. Purify the residue by C18
reversed phase
chromatography using a gradient of 30 to 73% 1:1 ACN:Me0H in 25 mM aqueous
ammonium carbonate solution to give the title compound. ES-MS m/z 591 (M+H).
Example 15
(S)-2-454-Cyano-16-methy1-3,9-dioxa-1,2(1,3),5(1,2)-tribenzenacyclononaphane-
14-
yOmethyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic
acid
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0
I¨ I
NC =.õ
N
0 N CO2H
0
Prepare the title compound essentially as described in Example 4 using methyl
(S)-24(54-cyano16-methy1-3,9-dioxa-1,2(1,3),5(1,2)-tribenzenacyclononaphane-14-
yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate.
Upon completion, concentrate the reaction mixture to one quarter volume,
neutralize with
citric acid solution and purify via C18 reversed phase chromatography eluting
with a
gradient for 10 to 80% ACN in 10 mM aqueous ammonium bicarbonate containing
5% Me0H to give the title compound. ES-MS ni/z 630 (M+H).
Example 16
(S)-4-Methoxy-24(16-methy1-54-(trifluoromethyl)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid
F3C
0 N N CO2 H
0
Prepare the title compound essentially as described in Example 1 using methyl
(S)-4-methoxy-2-016-methy1-54-(trifluoromethyl)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-
benzord]imidazole-6-carboxylate, using dioxane:ACN:water (6:6:1) as solvent,
and
stirring the reaction at 50 C for 2h. When complete, neutralize with citric
acid and
concentrate the reaction mixture. Dilute the residue with Et0Ac and wash with
water and
saturated aqueous NaCl. Dry the organic phase over sodium sulfate, filter, and
concentrate. Purify the residue via silica gel chromatography using a gradient
of 10 to
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80% (20% Me0H in Et0Ac) in DCM to give the title compound. ES-MS m/z 674
(M+H)_
Example 17
(S)-2-054-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-y1)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid
NC
011 0
0 N N ip
Prepare the title compound essentially as described in Example 2 using methyl
(S)-2-((54-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate. Stir the reaction at 45 'V for 4 h. Cool the
mixture to
RT, filter, and evaporate the filtrate. Purify the residue by reversed phase
chromatography
using a gradient of 30 to 73% solvent B in solvent A (solvent A = [65mM NH40Ac
+
ACN (90: I0)]; solvent B = ACN]) to give the title compound as a white solid.
ES-MS m/z
621 (M+H).
Example 18
(5)-24(54-Bromo-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
B r
0 N N CO2 H
Prepare the title compound essentially as described in Example 3 using methyl
(S)-2-454-bromo-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate and using
2:1
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THF:Me0H as solvent. Stir at 65 'V for 1.5 hand add 1 M aqueous KH2PO4. Dilute
the
reaction 2.5-fold with water and leave to cool with stirring for 45 min.
Collect the solid
by filtration and wash with 1:3 MeOH:water followed by water. Dry the filter
cake under
reduced pressure at 50 C for 20 h to afford the title compound as a white
solid. ES-MS
m/z 640 and 642 (M+H).
Example 19
(S)-2-((54-Cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
NC 0
=
r" 0
0 CO2H
I
Prepare the title compound essentially as described in Example 1 using methyl
(S)-24(54-
cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)methyl)-
1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate in 3:1:1 ACN : THF :
water.
Heat the mixture at 55 C for 3 h, cool to RT and quench with 5% aqueous
citric acid up
to = 4-5 to precipitate a white solid. Filter the solid, wash with water (3
times) and
ACN, and dry under vacuum at 45 C overnight to give the title compound as a
white
solid. ES-MS m/z 587 (M+H).
Example 20
(S)-2-((54-Cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
NC
f" 0
0 NN CO2H
I F
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Prepare the title compound essentially as described in Example 1 using methyl
(S)-2-((54-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacycl ooctaphane-14-yl)m ethyl )-1-(oxetan-2-ylm ethyl)-1H-benzo[d]i
mi dazol e-6-
carboxylate in 3:1:1 ACN : 1,4-dioxane : water as solvent. Heat the reaction
to 60 C for
3 h, then cool to RT and quench with a solution of citric acid (5% in water).
Dilute with
Et0Ac, separate the phases and extract the aqueous phase twice with Et0Ac.
Combine
the organic phases, wash with water and saturated aqueous NaCl, dry over
Na2SO4, filter
and concentrate under reduced pressure. Purify the residue by SFC [column:
Chiralpak
20x250mm, 5 m; isocratic mobile phase: 35 % CO2 in (Me0H + 0.5%
dimethylethylamine) at 100 bar, flow rate 65 mL/min] to give the title
compound as a
white solid. ES-MS m/z 591 (M+H).
Example 21
(S)-2-((54-Cyano-3, 8-dioxa-2(2,6)-pyridina-1,5(1,3)-dibenzenacyclononaphane-
14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
C N
0 )
0 N
N ip
co,H
Prepare the title compound essentially as described in Example 1 using methyl
(S)-2454-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (90 mg,
0.14
mmol) in 3:1:1 ACN : 1,4-dioxane : water as solvent. Heat the reaction to 60 C
for lh,
cool to RT and quench with aqueous citric acid (5%). Filter the solid and wash
with
water and then ACN to provide the title compound as a white solid. ES-MS m/z
587
(M+H).
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Example 22
(S)-4-Methoxy-2-((16-methyl-54-(fluoro)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacycl ononaphane-14-yl)methyl)-1-(oxetan -2-ylm ethyl)-1H-benzo[d]imi
dazol e-6-
carboxylic acid
f"' 0
0 N N c02H
--- 0
Prepare the title compound essentially as described in Example 4 using methyl
(S)-4-methoxy-2-016-methy1-54-(fluoro)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate, stirring the reaction at 50 C for 2h. Concentrate the reaction,
neutralize
with aqueous citric acid and purify via C18 reversed phase chromatography
using a
gradient of 10 to 80% ACN in 10 mM aqueous NH4HCO3 containing 5% Me0H. ES-
MS 111/Z 624 (M+H).
Example 23
24(54-Cy ano-3 ,9-dioxa-2(2,6)-pyridina-1(1,3),5 (1,2)-dibenzenacyclononaphane-
14-
yl)methyl)-4-methoxy-1-(oxazol-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic
acid
r&Nic-0
NC
0 N 10 CO2H
¨0
Prepare the title compound essentially as described in Example 1 using ethyl 2-
((54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)methyl)-4-methoxy-1-(oxazol-2-ylmethyl)-1H-benzokliimidazole-6-carboxylate,
stirring the reaction at 65 C for 72 h. Concentrate the mixture to one-
quarter volume and
adjust to pH = 5 using formic acid. Collect the resulting precipitate and
purify via C18
reversed phase chromatography eluting with a gradient for 10 to 80% ACN in 10
mM
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aqueous NH4HCO3 containing 5% Me0H to give the title compound as a colorless
solid.
ES-MS in/z 628 (M+H).
Example 24
(S)-2-((54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylic acid
NC
0 NN CO2H
0
0
Prepare the title compound essentially as described in Example 1 using methyl
(S)-2454-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-
14-
yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylate using 5:5:1 1,4-dioxane : ACN : water as solvent and stirring the
reaction at
45 C for 16 h. Concentrate the mixture to one-quarter volume and adjust to pH
= 5 using
formic acid. Dilute the mixture with water and extract the organics with
chloroform/isopropanol (3:1). Dry the organics over MgSO4, filterer and
concentrate.
Purify the title compound via flash chromatography eluting with a gradient of
0 to 40%
(10% formic acid in Me0H) in DCM. ES-MS in/z 661 (M+H).
Example 25
(S)-2454-(Hydroxymethyl)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacycl ononaphane-14-yl)methyl)-1-(oxetan -2-ylm ethyl )-1H-benzo[d] i
mi dazol e-6-
carboxylic acid
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HO --' , =0
f". 0
0 N 4.4 CO2H
Add sodium borohydride (5.3 mg, 0.14 mmol) to a slurry of (S)-2-((54-formy1-
3,9-
dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-
(oxetan-2-
ylmethyl)-1H-benzordlimidazole-6-carboxylic acid (30 mg, 0.051 mmol) in a
mixture of
Me0H (460 L) and TELF (1 mL) at 0 C. Stir mixture for 5 min and then warm up
to RT.
Stir reaction for 20 min and then remove volatiles with a nitrogen stream at
RT. Add
aqueous citric acid (5%), stir for 5 min, filter the solid and wash with water
and Me0H.
Purify the solid via silica gel chromatography using 10% of Me0H in DCM to
provide
the title compound (8 mg, 25%) as a white solid. ES-MS m/z 592 (MAT).
Example 26
(S)-4-Methoxy-2-((16-methy1-56-(trifluoromethyl)-3,9-dioxa-2(2,6),5(3,2)-
dipyridina-
1(1,3)-benzenacyclononaphane-14-y1)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid;1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-
a]pyrimidine
7
0 N N CO2 H
N
0
Prepare the title compound essentially as described in Example 4 using methyl
(S)-4-methoxy-2-016-methy1-56-(trifluoromethyl)-3,9-dioxa-2(2,6),5(3,2)-
dipyridina-
1(1,3)-benzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate, stirring the reaction at 45 C for 16 h.
Neutralize the
reaction with aqueous citric acid, concentrate the mixture, and purify via C18
reversed
phase chromatography using a gradient of 10 to 80% ACN in 10 mM aqueous
NH4HCO3
containing 5% Me0H. ES-MS m/z 675 (M+H).
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Example 27
(S)-4-Methoxy-2416-methyl-3,9-dioxa-2(2,6),5(4,3)-dipyridina-1(1,3)-
benzenacycl ononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-11-1-benzo[d]im
idazol e-6-
carboxylic acid
is"' 0
0 N OP CO2 H
I
0
Prepare the title compound essentially as described in Example 4, using methyl
(S)-4-methoxy-2-016-methyl-3,9-dioxa-2(2,6),5(4,3)-dipyridina-1(1,3)-
benzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-
6-
carboxylate, stirring the reaction at 50 C for 2h. Concentrate the reaction,
neutralize
with aqueous citric acid and purify via C18 reversed phase chromatography
using a
gradient of 10 to 80% ACN in 10 mM aqueous NH4HCO3 containing 5% Me0H.
Redissolve the purified product in DCM and neutralize with aqueous citric
acid. Wash
the organics with water and saturated aqueous NaCl. Dry over Na2SO4, filter
and
concentrate to give the title compound. ES-MS m/z 607 (M+H).
Example 28
(5)-1-(Oxetan-2-ylmethyl)-2-454-(1-(oxetan-3-ylmethyl)-1H-pyrazol-4-y1)-3,9-
dioxa-
2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-y1)methyl)-1H-
benzo[d]imidazole-6-carboxylic acid
\jµ
0 N N CO2 H
-
Add DMF (0.43 mL) and tripotassium phosphate (1 M aqueous solution, 0.13 mL,
0.13 mmol) to a vessel containing (S)-2-054-bromo-3,9-dioxa-2(2,6)-pyridina-
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1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid (30 mg, 0_0436 mmol), 1-(oxetan-3-
ylmethyl)-4-
(tetramethy1-1,3,2-dioxaborolan-2-y1)-11-/-pyrazole (19 mg, 0.0698 mmol) and
1,1'-bis(di-
tert-butylphosphino)ferrocene palladium dichloride (1.4 mg, 0.0021 mmol).
Purge the
vessel with nitrogen, seal the vessel, and stir the mixture at 60 C for 2 h.
Cool to
ambient temperature, and then purify the mixture directly via C18 reversed
phase
chromatography using ACN/10 mM aqueous Na4HCO3 as eluent.
Combine with material from a second analogous reaction, suspend the resulting
solid 1:1 DCM : Et0Ac, and partially concentrate under reduced pressure to
remove
DCM. Stir the suspension for 10 min at ambient temperature, then collect the
solid by
filtration and wash with Et0Ac. Dry under reduced pressure at 50 C for 16 h
to afford
28 mg of the title compound (average 41% for each of two reactions) as a white
solid.
ES-MS m/z 698.
Example 29
(S)-2454-(6-Methoxypyridin-3 -y1)-3 ,9-dioxa -2(2,6)-pyridina-1(1,3), 5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzordlimidazole-6-
carboxylic acid
0
NI
0 N NI = CO2H
Add (S)-2-((54-bromo-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid (30 mg, 0.0436 mmol), 2-methoxy-5-(4,4,5,5-tetramethy1-1,3,2-
dioxaborol an-2-yl)pyri dine (22 mg, 0.094 mmol) and 1,1'-bis(di-tert-
butylphosphino)ferrocene palladium dichloride (3 mg, 0.0045 mmol) into a glass
tube
with stir bar. Purge the tube with nitrogen and add tripotassium phosphate (1
M aqueous
solution, 0.13 mL, 0.13 mmol) and DMF (0.5 mL). Stir the mixture at 60 C for
16 h.
Add further 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (3
mg, 0.0045
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mmol) to the reaction and heat for 3 h at 60 C, then for 16 h at 90 C. Cool
the mixture
to ambient temperature, and then purify the reaction mixture directly using
C18 reversed
phase chromatography using ACN/10 mM aqueous NI-14HCO3 as eluent to give the
title
compound (4.7 mg, 14%) as a solid. ES-MS in/z 669.
The following examples are prepared essentially as described in Example 29
using
the appropriate boronic acid or boronate ester.
Example Name Structure
Characterization
30 (S)-2454-(1H-Pyrazol-4- N
H
ES-MS rn/z 628
y1)-3,9-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
=
14-yl)methyl)-1-(oxetan-2-
/N
ylmethyl)-1H-
benzo[d]imidazole-6- 40 0
carboxylic acid
N--
N-,
02 H
31 (S)-24(54-( 1-Methyl - 1H- N
ES-MS nilz 642
pyrazol-5-y1)-3,9-dioxa-
2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)methyl)-1-(oxetan-2-
/
ylmethyl)-1H- 40 0
benzo[d]imidazole-6-
carboxylic acid
N-,
02 H
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32 (S)-2454-(2-Methy1-2H-
xrµL-N1
ES-MS m/z 643
1,2,3-triazol-4-y1)-3,9-
¨N
dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
0
dibenzenacyclononaphane-
14-yl)methyl)-1-(oxetan-2-
ylmethyl)-1H- 1100 0
benzo[d]imidazole-6-
carboxylic acid N--
N--,
c5
02H
33 (S)-2454-(4- õ..;0
ES-MS m/z 716
S-
(Methylsulfonyl)pheny1)- sO
3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-
0
14-yOmethyl)-1-(oxetan-2-
ylmethyl)-1H- /N1
benzo[d]imidazole-6-
0
carboxylic acid
N--
Cb
02H
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34 (5)-245442- ¨0
ES-MS m/z 669
Methoxypyridin-4-y1)-3,9-
\ /
dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane- 0
14-yl)methyl)-1-(oxetan-2-
\ /
ylmethyl)-1H-
benzo[d]imidazole-6- 4.0 0
carboxylic acid
N__--
N,
Cb
02H
35 (S)-2-454-(1-Cyclopropyl- NJ
ES-MS m/z 668
õ4
1H-pyrazol-4-y1)-3,9-
dioxa-2(2,6)-pyridina-
=
1(1,3),5(1,2)-
0
dibenzenacyclononaphane-
14-yl)methyl)-1-(oxetan-2- \ /N
ylmethyl)-1H-
0
benzo[d]imidazole-6-
carboxylic acid =N.
02H
02H
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36 (S)-2454-(1-Methy1-6- \ 0
ES-MS m/z 669
oxo-1,6-dihydropyridin-3-
\ /
y1)-3,9-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane- 0
14-yl)methyl)-1-(oxetan-2-
ylmethyl)-1H-
benzo[d]imidazole-6- 110, 0
carboxylic acid
N__--
02 H
37 (5)-1-(Oxetan-2-y1methy1)- N_
ES-MS m/z 639
2-454-(pyridin-3-y1)-3,9- \ /
dioxa-2(2,6)-pyridina-
=
1(1,3),5(1,2)- 0
dibenzenacyclononaphane- ¨
14-yl)methyl)-1H- \ /
benzo[d]imidazole-6- 410 0
carboxylic acid
N__--
02 H
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38 (5)-245444-
ES-MS m/z 695
HN
(Methylcarbamoyl)pheny1)- 0
3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)methyl)-1-(oxetan-2- 0
ylmethyl)-1H-
/
benzo[d]imidazole-6-
carboxylic acid 1104 0
1.1
02H
39 (S)-2454-(4- HO
ES-MS m/z 668
(Hydroxymethyl)pheny1)-
3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)methyl)-1-(oxetan-2-
ylmethyl)-1H- /N
benzo[d]imidazole-6-
carboxylic acid 0
cb
02H
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76 (S)-1-(Oxetan-2-ylmethyl)-2- 0
ES/MS m/z 684
((54-(1-(oxetan-3-y1)-1H-
(M+H)
N'
pyrazol-4-y1)-3,9-dioxa-
N
\ /
2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacy clononaphane-1 4-
=
yOmethyl)-1H- 0
benzo[d]imidazole-6-
carboxylic acid \
= 0
HO 0
77 (S)-2454-(4-
ES/MS m/z 709
¨N
(Dimethylcarbamoyl)pheny1)- 0 (1\4+1-1)
3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacy clononaphane-1 4-
yl)methyl)- 1 -(oxetan-2-
0
ylmethyl)-1H-
benzo[d]imidazole-6- /N
carboxylic acid
11
N--
N--
opi
H 0 0
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78 (S)-24(54-(2-Aminopyridin- H2N ES/MS
m/z 654
, N
4-y1)-3,9-dioxa-2(2,6)- / \ (M+H)
pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-
0
ylmethyl)-1H-
/N
benzo[d]imidazole-6-
carboxylic acid
0
N¨
.
HO 0
79 (S)-2-((54-(1-Methy1-2-oxo-
, N/ ES/MS
m/z 669
1,2-dihydropyridin-4-y1)-3,9- 0 (1\4+1-
1)
dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14- 0
yl)methyl)- 1 -(oxetan-2-
ylmethyl)-1 /N
benzo[d]imidazole-6- = 0
carboxylic acid
N.¨
N,
H 0 0
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80 (5)-1-(Oxetan-2-ylmethyl)-2- 0 ES/MS
m/z 655
((54-(6-oxo-1,6- (M+H)
\ /
dihydropyridin-3-y1)-3,9-
dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
0
dibenzenacyclononaphane-14-
yl)methyl)-1H- \ IN
benzo[d]imidazole-6- = 0
carboxylic acid
N¨
N---
0
H 0 0
Example 40
(5)-2454-(1-Methy1-1H-pyrazol-4-y1)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
=
0 N
,
410 CO2 H
N
Prepare the title compound essentially as described in Example 29 using 1-
m ethyl -444,4, 5, 5-tetram ethyl -1,3,2-di oxaborol an -2-y1 )pyrazol e,
stirring the reaction at
60 C for 2 h and omitting the second addition of catalyst. Load the crude
reaction
mixture onto hydrophobic lipophilic balance (HLB) resin and elute with 10 mM
aqueous
NH4HCO3 buffer then with 1:1 DCM : Me0H. Concentrate fractions containing the
title
compound and then further purify using C18 reversed phase chromatography using
a
gradient of ACN in 10 mM aqueous NH4HCO3. ES-MS In/z 642.
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Example 41
(S)-144(1-(Oxetan-2-ylmethyl)-6-(1H-tetrazol-5-y1)-1H-benzo[d]imidazol-2-
y1)methyl)-
3,9-di oxa-2(2,6)-pyri di na-1(1,3),5(1,2)-dibenzenacycl ononaphane-54-
carbonitrile
N--"N
To a solution of (S)-1441-(oxetan-2-ylmethyl)-6-(1-((2-
(trimethyl silyl)ethoxy)methyl)-1H-tetrazol -5 -y1)-1H-benzo[d]imidazol-2-
yl)methyl)-3, 9-
dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-54-carbonitrile
(100 mg,
0.1 mmol) in THF (2 mL) and add TBAF (1 M in THF, 0.3 mL, 0.3 mmol). Stir the
mixture at 60 C for 16 h. Quench the reaction with water and dilute with
Et0Ac Dry the
organic phase over Mg SO4, filter, and concentrate under reduced pressure.
Suspend the
residue in an aqueous ammonia solution and stir at 50 C for 6 h. Concentrate
under
reduced pressure and purify the solid by C18 reversed phase chromatography
using a
gradient of 30 to 70% ACN in aqueous ammonium acetate solution to give 93 mg
of the
title compound (10%). ES-MS m/z 611 (M+H).
Example 42
(S)-24(54-Cyano-16-fluoro-3,6-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
NC 0
0 N F N = CO2H
To a solution of methyl (S)-24(54-cyano-16-fluoro-3,6-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclooctaphane-14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate (92 mg, 0.15 mmol) in ACN (8.4 mL) and water
(4.8
mL), add 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2-a]pyrimidine (87 mg, 0.61
mmol). Stir
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at 45 C overnight. Cool the mixture to RT, dilute with water and extract
three times with
3:1 DCM: isopropanol. Combine the organics, wash with water and saturated
aqueous
NaC1, dry over MgSO4, filter, and concentrate under reduced pressure. Purify
the residue
via silica gel chromatography using 30% Et0Ac in DCM and then a gradient of 0
to 5%
(10:1 Me0H : formic acid) in DCM. Repurify by reverse phase chromatography
using 41
to 83% [1:1 ACN : MeOH] in 0.1% aqueous formic acid (pH3) to give the title
compound
(12 mg, 13%) as a white solid. ES-MS m/z 591 (M+H).
Example 43
(S)-24(55-Cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-dipyridina-1(1,3)-
benzenacyclononaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid diformate salt
NC
, 0
0
0 0 N N 4100 CO2H
H F
Me
0
(0 H
To a suspension of methyl (S)-2-((55-cyano-16-fluoro-3,9-dioxa-2(2,6),5(2,3)-
dipyridina-1(1,3)-benzenacyclononaphane-14-yl)methyl)-4-methoxy-1-(oxetan-2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (66 mg, 0.10 mmol) in ACN (1.2
mL),
THF (0.3 mL) and water (0.2 mL), add 1,3,4,6,7,8-hexahydro-2H-pyrimido [1,2-
a]pyrimidine (0.032 g, 0.23 mmol). Stir the suspension at 45 C for 6 h. Cool
the mixture
to RT, add formic acid until pH = 4, and extract with Et0Ac (3 > 5 mL).
Combine the
organics, dry over MgSO4, filter, and concentrate under reduce pressure.
Purify the
residue via silica gel chromatography using a gradient of 0 to 50% solvent B
in solvent A,
wherein solvent B = (DCM / Me0H / formic acid 9:0.9:0.1) and solvent A = DCM
to
give the title compound as a beige solid (15 mg, 24%). ES-MS 112/Z 636 (M+H-
formate
salt).
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Example 44
(S)-24(54-Cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-y1 )methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid
NC
0 /"µ 0
/ 0
0 Ist., N
I F 0 H
¨0
To a suspension of methyl (S)-2454-cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-4-methoxy-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate (57 mg, 0.09 mmol) in degassed ACN (0.9
mL),
1,4-dioxane (0.3 mL) and water (0.3 mL), add 1,5,7-triazabicyclo[4.4.0]dec-5-
ene (38
mg, 0.27 mmol). Heat the reaction mixture at 60 C for 2 h, cool to RT, add 5%
aqueous
citric acid solution up to pH = 5, then add water (2.0 mL) and stir the
mixture at RT for
min. Filter the resulting solid, wash with water (5 mL) and dry under vacuum
at 45 C
overnight. Suspend the solid in Me0H (1.0 mL) and stir the mixture for 15 min
at RT.
Filter the resulting solid, wash with Me0H (0.5 mL), Et0Ac (1.5 mL) and dry
under
15 vacuum at 45 C overnight to give the title compound as a pale
brown solid (19 mg,
34%). ES-MS nilz 635.2/636.2 (M+H).
Example 45
(S)-2454-Chloro-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid
c 0
=C)
F 0 N N CO2H
F 0
0¨
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To a solution of methyl (S)-24(54-chloro-16,56-difluoro-3,8-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-4-(2-methoxyethoxy)-1-
(oxetan-2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (245 mg, 64% purity, 0.21 mmol)
in
ACN (2.4 mL), water (0.8 mL) and THF (0.8 mL) at 55 C, add 1,5,7-
triazabicyclo
[4.4.0]dec-5-ene (210 mg, 1_47 mmol). Stir the mixture at 55 C for 2 h, cool
to RT, add
5% aqueous citric acid until pH = 4-5, then filter the white solid which
precipitates.
Dissolve the solid in ACN/Me0H and purify by preparative HPLC [column: Welch
Xtimate C18 150 x 30 mm x 5 pm; mobile phase: 30 to 70% ACN in aqueous formic
acid (0.225%)] to afford the title compound as a white solid (31 mg, 20%). ES-
MS nilz
706 (M+H).
Example 46
(S)-2-((55-Cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-benzenacyclononaphane-14-
yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylic acid
CN
N
N
CO2H
N 110
0
Prepare the title compound essentially as described in Example 5 using methyl
(S)-2-((55-cyano-3,8-dioxa-2,5(2,6)-dipyridina-1(1,3)-benzenacyclononaphane-14-
yl)m ethyl )-4-(2-m eth oxyethoxy)-1-(oxetan -2-ylm ethyl )-1H-benzo[d]i m
dazol e-6-
carboxylate (63 wt% pure) in a mixture of ACN: THF : water (3:1:1), heating
the
reaction at 55 'V under nitrogen atmosphere for 2h. Cool the reaction mixture
to RT and
quench with aqueous citric acid (5%), then filter the solid and wash with
water. Purify by
preparative HPLC [column: Welch Xtimate C18 150 >< 30 mm, 5 p.m; mobile phase:
25 to
65% ACN in aqueous formic acid (0.225%)] to give title compound as a white
solid. ES-
MS in/z 662 (M+H).
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Example 47
(S)-2-054-Chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid
CI
0
OJyiJ N CO2 H
I F
0
0
Prepare the title compound essentially as described in Example 2 using methyl
(S)-2-((54-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-
14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylate in 10:5:3 ACN : 1,4-dioxane : water as solvent, stirring the
reaction mixture
at 55 C for 6 h 30 min. Concentrate the crude reaction mixture onto Centel')
and purify
by C18 reversed phase chromatography using a gradient of 10 to 73% ACN in
aqueous
NH4HCO3 (10 mM plus 5% Me0H) to give the title compound. ES-MS nilz 688 (M+H).
Example 48
(S)-2451-Cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-
14-y1 )methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]i midazole-
6-
carboxylic acid
C N
010 0
=
= 0
0 INt., N CO2 H
I F
0
0
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To a mixture of (S)-24(54-chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid (15 mg, 0.022 mmoL), potassium
ferrocyanide
trihydrate (14.5 mg, 0.039 mmoL), XPhos Pd(crotyl)C1 (Pd-170, 5.5 mg, 0.008
mmoL),
and KOAc (5.6 mg, 0.056 mmoL) add 1,4-dioxane (1.0 mL) and water (0.4 mL).
Stir the
mixture at 90 C for 4 h. Concentrate the reaction mixture onto Celitec' and
purify by C18
reversed phase chromatography using a gradient of 10 to 73% ACN in aqueous
NH4HCO3 (10 mM plus 5% Me0H) to give 8.2 mg of the title compound (55%). ES-MS
nilz 679 (M+H).
Example 49
(S)-2-454-Cyano-23-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid
NC 0
0 N...õN CO2H
I
F 0
0
Prepare the title compound essentially as described in Example 45 using methyl
(S)-24(54-cyano-23-fluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate. Purify by preparative HPLC [column: Welch
Xtimate C18 150 >< 25 mm, 5 lam; mobile phase: 25 to 70% ACN in aqueous TFA
(0.1%)]. Remove the organic solvent under reduced pressure and lyophilize the
residual
aqueous solution to get the titled product as a white solid. ES-MS nilz 679
(M+H).
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Example 50
(S)-24(54-cyano-6,6-difiuoro-3.8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacycl onaph ane-14-yl)m ethyl)-4-(2 -methoxyethoxy)-1 -(oxatan -2-ylm
ethyl)-1 H-
benzo[d]imidazole-6-carboxylic acid
F F
NCIIr
0
=
0
0 N N
0 H
0
0 ¨
Dissolve methyl (S)-2454-cyano-6,6-difluoro-3.8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclonaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-
(oxatan-2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (61 mg, 0.086 mmol) in ACN (1
mL),
1,4-di oxane (0.3 mL), and water (0.3 mL). To this solution add 1,5,7-
triazabicyclo[4.4.0]clec-5-ene (0.04 g, 0.28 mmol) and stir this mixture at
ambient
temperature for 18 h. After this time, quench the reaction with 1N HCl (to pH
5), and
extract with Et0Ac. Dry the organics over MgSO4, filter, and concentrate.
Purify this
material with reverse phase HPLC [column: Phenomenex Kinetex EVO C18 100 x 30
mm, 5 pm; mobile phase: 23 to 58% ACN in aqueous NH4HCO3 (10 mM plus 5%
Me0H)] to give the title compound as a white solid (14.5 mg, 24.1%). ES-MS
(in/z)
697.4 (M+H).
Example 51
(S)-2-((54-Cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-
dibenzenacyclononaphane-
14-yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylic acid
NC 0
is 0
I / 0
0 N N
IN 0 H
0
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Prepare the title compound essentially as described in Example 2 using methyl
(S)-24(54-cyano-3,8-dioxa-2(2,4)-pyrimidina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-methoxy-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxyl ate
as
starting material, using 5:2:1 ACN : 1,4-dioxane : water as solvent, heating
the reaction to
40 C for 21 h. Upon completion, cool the reaction to ambient temperature and
quench
with 5% aqueous citric acid bringing the pH to 4. Filter the resulting
precipitate and wash
the solid with water. Dry the collected solid under reduced pressure to give
the title
compound. ES/MS m/z 618 (M+H).
Example 52
(S)-4-(2-Methoxyethoxy)-1-(oxetan-2-ylmethyl)-2-((56-(trifluoromethyl)-3,8-
dioxa-
2(2,6),5(3,4)-dipyridina-1(1,3)-benzenacyclononaphane-14-y1)methyl)-1H-
benzo[cilimidazole-6-carboxylic acid
F
N I
0 H
0 N N =
0
0
0
Prepare the title compound essentially as described in Example 2 using methyl
(S)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-24(56-(trifluoromethyl)-3,8-
dioxa-
2(2,6),5(3,4)-dipyridina-1(1,3)-benzenacyclononaphane-14-y1)methyl)-1H-
benzordlimidazole-6-carboxylate and 2.8:1:1 ACN : THF : water as solvent,
heating the
reaction at 45 C for 2 h. Quench the reaction with 1 M aqueous citric acid
bringing the
pH to 4.5. Filter the resultant colorless solid and dry under vacuum. Purify
via reverse
phase chromatography on a C18 column using a gradient of 42% to 75% ACN in
aqueous
formic acid (0.225%) to give the title compound. ES/MS nilz 705 (M+H).
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Example 53
(S)-2-((54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)m ethyl )-4-(2-(2-m ethyl oxazol -4-yl)ethoxy)-1-(oxetan-2-ylmethyl)-11-1-
benzo[d]imidazole-6-carboxylic acid
NC
001 0
r"
, 0
0 N NNe,
f
,
0 H
0
N
Prepare the title compound essentially as described in Example 5 using methyl
(S)-24(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-(2-(2-methyloxazol-4-ypethoxy)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate and using mixture of 3:1:1 ACN : THY : water
as
solvent, stirring the reaction at 35 C for 6 h. Upon completion, concentrate
the mixture
and dissolve the residue in a minimum of DMSO. Filter the DMSO solution and
purify
the filtrate by preparative HPLC [column: Welch Xtimate C18 150 x 30 mm, 5
p.m;
mobile phase: gradient of 10 to 45% ACN in aqueous NH4HCO3 (10mM) to give the
title
compound as a white solid. ES/MS miz 712 (M+H).
Example 54
(S)-2-((54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-4-(2-((2,2,2-trifluoroethyl)amino)ethoxy)-1H-
benzo[d]imidazole-6-carboxylic acid
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NC
0
r' o
, 0
,
OH
0
F
N H
To a solution of (S)-4-(2-((tert-butoxycarbonyl)(2,2,2-
trifluoroethyl)amino)ethoxy)-2-
((54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
(Preparation
311, 75 mg, 0.058 mmol) in toluene (7 mL), add silica gel (750 mg), and heat
the mixture
to 120 C for 18 h. Cool the mixture to ambient temperature, filter, and
concentrate in
vacuo. Purify the residue via preparative HPLC [column: Xtimate C18 100 x 30
mm, 10
um; mobile phase: gradient of 35 to 65% ACN in aqueous formic acid (0.2%)] to
give the
title compound as a colorless solid (2.5 mg, 5.7%). ES/MS in/z 728.6 (MAI).
Example 55
(S)-442-Hydroxyethoxy]-2-454-chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
CI 0
0
N
I OH
0
0 H
To a solution of methyl (S)-442-[tert-butyl(dimethypsilylioxyethoxy]-2454-
chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-
yOmethyl)-
1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (62 mg, 0.079 mmol)
in THE
(5 mL) and Me0H (2 mL), add LiOH (10 mg, 0.42 mmol) dissolved in water (2 mL)
and
heat to 45 'V for 1.5 h. Add more LiOH (12 mg, 0.050 mmol) dissolved in water
(1 mL)
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and heat for another hour at 45 'V, cool to ambient temperature, and
concentrate under
reduced pressure. Suspend the crude material in water (20 mL) and adjust the
pH to 5
with 1N HC1. Filter the solid, collect, and dry under reduced pressure. Purify
via C18
reversed phase chromatography eluting with 30-60% ACN in 10mM aqueous ammonium
bicarbonate containing 5% Me0H to give the title compound (7.5 mg, 14%).
ES/MS (nilz): 656.4 (M+H).
Example 56
(S)-2-((54-Chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-(2-(dimethylamino)ethoxy)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
CI 0
NI""=
0
0 N N
0 H
0
N¨
/
To a nitrogen-spargedmixture of methyl (S)-2-054-chloro-3,8-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-4-(2-
(dimethylamino)ethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylate (89
mg, 0.12 mmol) in ACN (2 mL), 1,4-dioxane (1 mL) and water (0.22 mL), add
1,5,7-
triazabicyclo[4.4.0]dec-5-ene (60 mg, 0.42 mmol). Heat reaction mixture at 60
C for 16
h. Cool the mixture to ambient temperature, concentrate under reduced
pressure, and
purify the residue via reverse-phase chromatography using a gradient of 10 to
80% ACN
in water (0.1% formic acid added to both the ACN and water) to give the title
compound
as a colorless solid (42 mg, 48%). ES/MS m/z 683 (M+H).
Example 57
(S)-2-((54-Chloro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-(2-(methylamino)ethoxy)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
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-3 1 8-
C I 0
().
0
0 N
I 0 H
0
N H
To (S)-4-(2-((tert-butoxycarbonyl)(methyl)amino)ethoxy)-2-((54-chloro-3,8-
dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-
(oxetan-2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (12.3 mg, 0.016 mmol) in DCM
(2
mL), add TFA (0.1 mL, 1 mmol). Stir the mixture at ambient temperature for 15
min,
then concentrate the reaction under reduced pressure and purify residue by
reverse-phase
chromatography using a gradient of 10 to 90% ACN in water (0.1% formic acid
added to
both solvents) to give the title compound (2.2 mg, 21%). ES/MS (in/z). 669
(M+H)
Example 58
(S)-2454-Cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
dibenzenacyclononaphane-
14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylic acid
I I
0 CJO
0 N 0
0 H
0
0
Add 1,5,7-triazabicyclo[4.4.0]dec-5-ene (29 mg, 0.20mmol) to a degassed
suspension of methyl (S)-2-454-cyano-16-fluoro-3,8-dioxa-2(2,6)-pyridina-
1,5(1,3)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzordlimidazole-6-carboxylate (50 mg, 0.069 mmol) in a mixture of ACN
(1.4
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-3 1 9-
mL), 1,4-dioxane (0.5 mL) and water (0.5 mL). Heat the mixture at 60 C under
N2 for
1.5 h, cool to RT and quench with aqueous citric acid (5%). Filter the solid
and wash
with water and then ACN to provide the title compound (34 mg, 70%) as a
colorless
solid. ES-MS nilz 679 (M+H).
Example 59
24(54-Cy ano-3 ,9-dioxa-2(2,6)-pyridina-1 (1,3),5 (1,2)-
dibenzenacyclononaphane- 14-
yl)methyl)-4-((1-methylpyrrolidin-3-y1)oxy)-1-(((S)-oxetan-2-yl)methyl)-1H-
benzo[d]imidazole-6-carboxylic acid
0
0 N
,
0 H
0
t\N
Stir a solution of methyl 24(54-cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-((1-methylpyrrolidin-3-yl)oxy)-14(S)-
oxetan-2-yl)methyl)-1H-benzo[d]imidazole-6-carboxylate (90 mg, 0.12 mmol) and
1,5,7-
triazabicyclo[4.4.0]dec-5-ene (55 mg, 0.39 mmol) in a nitrogen-sparged mixture
of 1,4-
dioxane (0.4 mL), water (0.4 mL), and ACN (0.4 mL) at 25 C for 6 h. Adjust
the mixture
to pH 7 with formic acid and concentrate. Purify the residue via reversed
phase
chromatography using a gradient of 6 to 46% MeCN in aqueous ammonium hydroxide
(0.04%) plus NH4HCO3 (10 mM) to give 30.5 mg of the title compound (35%). ES-
MS
nilz 686 (M+H).
Example 60
(S)-2-454-Cyano-16,23-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid
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NC 0
f 0
0 N N CO2 H
,
F
0
0
To a degassed solution of methyl (S)-2-054-cyano-16,23-difluoro-3,8-dioxa-
2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-4-(2-
methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (250
mg,
0.16 mmol) in THF (1.17 mL), water (1.17 mL) and ACN (3.51 mL) add 1,5,7-
triazabicyclo[4.4.0]dec-5-ene (150 mg, 1.06 mmol). Seal the reaction vessel
and purge
with nitrogen. Heat the reaction mixture to 45 C and stir at this temperature
for 2 h.
Quench the reaction with 1 M aqueous citric acid until pH = 4.5, then filter
the colorless
solid and dry under reduced pressure. Purify the solid by preparative HPLC
(column:
Welch Xtimate C18 150 25 mm, 5 um), mobile phase: gradient of 25 to 60% ACN in
aqueous formic acid) to give the title compound as a colorless solid (53.9 mg,
49%) ES-
MS nilz 697 (M+H).
Example 61
(S)-2-((54-cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-benzord]imidazole-6-carboxylic
acid
N
0
t". 0
NI 0
0 N
0 H
Stir a solution of methyl (S)-2-454-cyano-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylate (20 mg, 0.03 mmol) and 1,3,4,6,7,8-hexahydro-
2H-
pyrimido[1,2-A]pyrimidine (20 mg, 0.1 mmol) in ACN (6 mL) and water (4 mL) at
45 C
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for 7 h. Adjust to pH 6 with formic acid and extract with 3:1
chloroform:isopropanol. Dry
the organic layer over magnesium sulfate, filter, and concentrate. Purify the
residue via
silica gel chromatography using a gradient of 0 to 10% (Me0H + 10% formic
acid) in
DCM to give 4 mg of the title compound (20%). ES-MS nilz 605 (M+H).
Example 62
(S)-2-((54-Cyano-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-(2-(dimethylamino)-2-oxoethoxy)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid
NC
N
1 10 0
0 N =
0 H
0
ON¨
/
Stir a solution of methyl (S)-2-054-cyano-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-(dimethylamino)-2-oxoethoxy)-1-
(oxetan-
2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (50 mg, 0.0356 mmol) with
trimethyltin hydroxide (34 mg, 0.178630 mmol) in 1,2-dichloroethane (1 mL) at
80 C for
16 h. Increase the temperature to 90 C for 7 h, and then to 100 C for 72 h.
Add
additional trimethyltin hydroxide (34 mg, 0.179 mmol) and heat to 105 C for 18
h.
Allow the reaction mixture too cool to ambient temperature and concentrate
under
vacuum. Add 15% aqueous citric acid (1 mL) to form a gum that is filtered
slowly over a
fritted funnel. Wash the gum with water (2 mL) and dry at 60 C in a vacuum
oven.
Purify the crude product by preparative HPLC [column: Phenomenex Kinetex EVO
C18
250 x 30 mm, 5 um; mobile phase: gradient of 0 to 100% ACN in aqueous N1-141-
1CO3 (10
mM + 5% Me0H)] to give the title compound as a colorless solid (12.5 mg, 51%).
ES-
MS nilz 688 (M+H).
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Example 63
24(54-Cy ano-3 ,9-dioxa-2(2,6)-pyridina-1(1,3),5 (1,2)-dib
enzenacyclononaphane-14-
yl)m ethyl)-1-((l-ethyl-1H-imi dazol -5 -yl)m ethyl)-4-methoxy-1H-
benzo[d]imidazole-6-
carboxylic acid
NN
,
0
0 N =, 0 H
0
Use nitrogen to sparge a solution of 1,5,7-triazabicyclo[4.4.0]dec-5-ene (28
mg,
0.197 mmol) in a mixture of ACN (0.4 mL), 1,4-dioxane (0.4 mL), and water
(0.15 mL)
for 10 min. Add the oxygen-free solution to a reaction vessel methyl 24(54-
cyano-3,9-
dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-1-((1-
ethyl-
1H-imidazol-5-yl)methyl)-4-methoxy-1H-benzo[d]imidazole-6-carboxylate (44 mg,
0.065 mmol). Stir the mixture at ambient temperature for 24 h. Partition the
reaction
mixture between Et0Ac and 0.1 M aqueous HC1. Separate the organic layer and
wash
with saturated aqueous NaC1, dry over MgSO4, filter, and concentrate under
reduced
pressure. Purify by the residue by preparative 1-IPLC [column: Phenomenex
Kinetex
EVO C18 100 x 30 mm, 5 Rm; mobile phase: gradient of 14 to 48% ACN in aqueous
formic acid (0.1%)] to afford the title compound (8.2 mg, 19%). ES-MS
171/27655 (M-4-1).
Example 64
(S)-2-054-Cyano-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-
yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylic acid
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NC 0
0
0 N N
0 H
0
0
To a suspension of methyl (S)-2-054-cyano-3,8-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-4-(2-methoxyethoxy)-1-
(oxetan-2-
ylmethyl)-1H-benzokl]imidazole-6-carboxylate (23 mg, 0.033 mmol) in degassed
ACN
(0.5 mL), THF (0.2 mL) and water (0.2 mL), add 1,5,7-triazabicyclo[4.4.0]dec-5-
ene (15
mg, 0.11 mmol). Heat the mixture to 55 C for 3 h, then cool to RT and add 5%
aqueous
citric acid to bring pH = 4-5. Filter the resulting solid, wash the solid with
water (3 times)
and ACN, then dry under vacuum at 45 C overnight to give the title compound
as a beige
solid (15 mg, 63%). ES-MS m/z 661 (M+H).
Example 65
(S)-2454-Cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-dibenzenacyclononaphane-14-
yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylic acid
I I
0
1411
N
0 N
NI 0
'OH
0
0
Add 1,5,7-triazabicyclo[4.4.0]dec-5-ene (26 mg, 0.18mmo1) to a degassed
suspension of methyl (S)-24(54-cyano-3,8-dioxa-2(2,6)-pyridina-1,5(1,3)-
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dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylate (45 mg, 0.067 mmol) in a mixture of ACN
(1.4
mL), 1,4-dioxane (0.5 mL) and water (0.5 mL). Heat the mixture at 60 C under
N2 for
1.5 h, cool to RT and quench with aqueous citric acid (5%). Filter the solid
and wash
with water and then ACN to provide the title compound (26mg, 59%) as a white
solid_
ES-MS m,/z 661 (M-41).
Example 66
(S)-2-((54-(4-Fluoro-1H-imidazol -1 -y1)-3 ,9-dioxa-2(2,6)-pyridina-1(1,3),
5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
0
0
0 NL, N =
I OH
Add aqueous LiOH (1M, 120 uL, 0.12 mmol) to a suspension of methyl (S)-2-
((54-(4-fluoro-1H-imidazol-1-y1)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-y1)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylate (30 mg, 0.045 mmol) in a mixture of Me0H (300 uL) and THF (600 pL)
and
stir at 60 C for 12 h. Concentrate the reaction mixture and add aqueous
citric acid (5%).
Filter off the solid, wash with water, ACN and Me0H to provide the title
compound (11
mg, 47%) as a pale brown solid. ES-MS m/z 646 (M-J-1).
Example 67
(S)-2-((54-Chl oro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacycl ooctaphan e-14-yl)m ethyl )-4-(2-meth oxy ethoxy)-1-(ox etan-2-
y1 methyl )-1H-
benzo[d]imidazole-6-carboxylic acid
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CI
0 õ
0 N N =
I F 0
Add aqueous LiOH (1 M, 1.6 mL, 1.6 mmol) to a suspension of methyl (S)-24(54-
chloro-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclooctaphane-14-
yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-
carboxylate (285 mg, 0.39 mmol) in a mixture of THF (8.5 mL) and Me0H (4.3
mL).
Heat the mixture at 60 C for 3 h. Concentrate the reaction mixture, add
aqueous citric
acid (5%), then filter off the solid and wash it with water and ACN. Dry the
solid under
vacuum at 40 C to obtain the title compound (285 mg, 100%) as a white solid.
ES-MS
m/z 674, 676 (M+H).
Example 68
(S)-2-((54-Cyano-16-fluoro-3,7-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacycl ooctaphane-14-yOmethyl)-4-(2-methoxy ethoxy)-1-(oxetan-2-
ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid
N
0 õ
0 N N =
,
0
I F
0
Charge a reaction vessel with (S)-2-454-chloro-16-fluoro-3,7-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclooctaphane-14-yl)methyl)-4-(2-
methoxyethoxy)-1-
(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (61 mg, 0.087
mmol),
tetrapotassium hexacyanoferrate trihydrate (57 mg, 0.135 mmol), KOAc (19 mg,
0.19
mmol) and chloro(crotyl)(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-
biphenyl)
palladium(II) (Pd-170, XPhos Pd(crotyl)C1, 6.2 mg, 0.009 mmol]. Purge the
reaction
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mixture with N2 and add DMF (1.7 mL) and water (0.9 mL). Purge the mixture
again
with N2 for 5 min, seal the vessel and stir at 90 C overnight. Cool the
mixture to RI, add
aqueous citric acid (5%), filter off the solid and wash with water and ACN.
Purify the
solid by preparative HPLC [column: )(Bridge C18 19 x 150 mm, 5 um, mobile
phase:
gradient of 30 to 50% ACN in aqueous NH4HCO3 (20 mM, pH9) to provide the title
compound (9 mg, 16%) as a white solid. ES-MS m,/z 665 (M+H).
Example 69
(S)-N-((Cyclopropylmethyl)sulfony1)-2-((54-fluoro-16-methyl-3,9-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-4-methoxy-1-
(oxetan-2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxamide
I / 0
0 N N
1 N¨S=0
0 H
To a solution of (S)-4-methoxy-24(16-methy1-54-(fluoro)-3,9-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid (100 mg, 0.16 mmol) in DCM (2 mL) add 4-
dimethylaminopyridine (20 mg, 0.16 mmol) cyclopropylmethanesulfonamide (35 mg,
0.26 mmol), TEA (0.1 mL, 0.7 mmol) and purge the mixture gently with N2 for 10
min.
Add EDC (50 mg, 0.26 mmol) to the reaction mixture, stir at RT overnight, then
heat the
reaction at 40 C for 4 h. Quench the reaction with aqueous citric acid (5%)
and dilute
with DCM. Separate the phases and the extract the aqueous layer twice with
DCM.
Combined the organic phases, wash them with saturated aqueous NaC1, dry over
Na2SO4,
filter off the solid by filtration and remove the solvent under reduced
pressure. Purify the
residue by preparative HPLC [column: Xbridge C18 150 x 19mm, 5 um, mobile
phase:
gradient of 50 to 80% ACN in aqueous NH4CO3 (20 mM, pH = 9)] to give the
titled
product as a white solid (28 mg, 23.6%) ES-MS m/z 741(M+H).
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Example 70
(S)-2-((54-Cyano-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid
N
0
F 0 N N = CO2 H
,
I F
0
0¨
Prepare the title compound essentially as described in Example 68 using (S)-2-
454-chloro-16,56-difluoro-3,8-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-4-(2-methoxyethoxy)-1-(oxetan-2-
ylmethyl)-
1H-benzo[d]imidazole-6-carboxylic acid as starting material, using a 2.5:1
mixture of
1,4-dioxane : water as solvent, heating the reaction to 90 C for 4 h. Purify
by preparative
HPLC [column: C18 100 x 30 mm, 5 pm; mobile phase: gradient of 15 to 85% ACN
in
aqueous formic acid (0.225%)] to afford the title compound as a white solid.
ES-MS m/z
697 (M+H).
Example 71
(S)-2-((54-(3-Fluoro-4-oxopyridin-1(4H)-y1)-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
0
FN 0
r 0
0
0 H
Charge a reaction vessel with (S)-24(54-bromo-3,9-dioxa-2(2,6)-pyridina-
1(1,3),5(1,2)-dibenzenacycl on onaphane-14-yl)m ethyl)-1 -(oxetan-2-ylm ethyl)-
1II-
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benzo[dlimidazole-6-carboxylic acid (51 mg, 0.079 mmol), 3-fluoropyridin-4-ol
(14 mg,
0.12 mmol), copper(I) iodide (1.6 mg, 0.008 mmol) and K2CO3 (22 mg, 0.156
mmol).
Purge the reaction vessel with N2, then add anhydrous DMSO (80 FL) and 2,2,6,6-
tetramethy1-3,5-heptanedione (7 uL, 0.033 mmol). Stir the reaction mixture at
120 C for
9 h. Cool the mixture to RT, add aqueous citric acid (5%), then filter off the
solid and
wash it with water. Purify by preparative HPLC [column: )(Bridge C18 19 150
mm, 5
um, mobile phase: gradient of 30 to 60% ACN in aqueous NH4HCO3 (20 mM, pH9)]
to
provide the title compound (10 mg, 18%) as a white solid. ES-MS nil z 673
(M+H).
Example 72
(S)-2-054-(4-Methy1-1H-imidazol -1 -y1)-3,9-dioxa-2(2,6)-pyri dina-
1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
= C')
1 0
0 N N
0 H
Add anhydrous DMSO (0.06 mL) to a mixture of (S)-2454-bromo-3,9-dioxa-
2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-y1)methyl)-1-(oxetan-
2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (20 mg, 0.031 mmol), 4-
methylimidazole (10 mg, 0.119 mmol), tripotassium phosphate (13 mg, 0.059
mmol),
N1,N2-bis(furan-2-ylmethyl)oxalamide (3 mg, 0.012 mmol) and copper(1) oxide (5
mg,
0.034 mmol). Purge the reaction vessel with argon, seal the vessel and heat to
120 C
with stirring for 19 h. Combine the reaction mixture in DCM (20 mL) with a
second
mixture prepared under similar conditions with bis(tetrabutylammonium
iodide)copper(I)
iodide (7 mg, 0.006 mmol) in place of copper(I) oxide. Add water (5 mL), 2-
propanol (5
mL), aqueous citric acid (5%, 5 mL) and saturated aqueous NaCl (20 mL), then
shake the
mixture and separate the phases. Extract the aqueous phase using 4:1 DCM: 2-
propanol
(50 mL in two portions) Adjust the aqueous pH to 3 using aqueous tripotassium
phosphate and extract again using 4:1 DCM : 2-propanol (15 mL). Combine the
organic
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extracts and concentrate under reduced pressure. Purify the residue by reverse-
phase
chromatography using a gradient of 30 to 60% ACN in aqueous NH4HCO3 (10 mM, pH
=
9) to give the title compound (6 mg, 15%) as a solid. ES-MS nilz 642 (M+H).
Example 73
(S)-2-454-(1H-Imidazol-1-y1)-3,9-dioxa-2(2,6)-pyridina-1(1,3),5(1,2)-
dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-
carboxylic acid
= C''.7
I"' 0
0 N N = 0
OH
Add anhydrous DMSO (0.20 mL) to a mixture of (S)-24(54-bromo-3,9-dioxa-
2(2,6)-pyridina-1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1-(oxetan-
2-
ylmethyl)-1H-benzoklimidazole-6-carboxylic acid (30 mg, 0.046 mmol), imidazole
(13
mg, 0.189 mmol), tripotassium phosphate (20 mg, 0.091 mmol), N1,N2-bis(furan-2-
ylmethyl)oxalamide (5 mg, 0.02 mmol) and copper(I) oxide (3 mg, 0.02 mmol).
Purge
the vessel with argon, seal the vessel and then heat to 120 C with stirring
for 20 h.
Transfer the reaction mixture to a Me0H-washed strong anion exchange resin
cartridge
(Isolute SAX) and elute sequentially with 7:3 Me0H/water, Me0H, DCM, and 1:1
DCM : Me0H with 3% acetic acid. Combine fractions containing the title
compound and
concentrate under reduced pressure. Dissolve the residue in DMSO and then
purify the
mixture by reverse-phase chromatography using a gradient of 30 to 60% ACN in
aqueous
NH4HCO3 (10 mM, pH = 9). Concentrate the appropriate fractions under reduced
pressure. Triturate the solid residue in 1:1 DCM : Et0Ac (4 mL) and partially
concentrate the mixture to remove DCM. Centrifuge the suspension, remove the
supernatant liquid and then dry the residue at 50 C under reduced pressure
for 24 h to
afford 9.4 mg of the title compound (29% yield) as a white solid. ES-MS iniz
628 (M+H).
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Example 74
(S)-1-(Oxetan-2-ylmethyl)-2-((54-(4-oxopyridin-1(4H)-y1)-3,9-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yl)methyl)-1H-benzo[d]imidazole-6-
carboxylic acid
0
0 -0
" 0
0 N N 0
0 H
Add anhydrous DMSO (80 L) and 2,2,6,6-tetramethy1-3,5-heptanedione (7 [IL,
0.033 mmol) to a nitrogen-purged mixture of (S)-2454-bromo-3,9-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclononaphane-14-yOmethyl)-1-(oxetan-2-ylmethyl)-1H-
benzo[d]imidazole-6-carboxylic acid (50 mg, 0.078 mmol), 4-hydroxypyridine (10
mg,
0.102 mmol), copper(I) iodide (1.6 mg, 0008 mmol) and K2CO3 (22 mg, 0.156
mmol).
Seal the vessel and stir at 120 C for 16 h. Cool the mixture to RT, add
aqueous citric acid
(5%), filter off the solid and wash it with water. Purify the solid by
preparative HPLC
[column: XBridge C18 19 x 150 mm, 5 Mm; mobile phase: gradient of 25 to 55%
ACN in
aqueous NH4HCO3 (20 mM, pH = 9)], then wash the solid product with Me0H and
ACN
to give the title compound (11 mg, 18%) as a colorless solid. ES-MS nilz 655
(M+H).
Example 75
(S)-24(16-Fluoro-54-(1-methyl-6-oxo-1,6-dihydropyridin-3-y1)-3,7-dioxa-2(2,6)-
pyridina-
1(1,3),5(1,2)-dibenzenacyclooctaphane-14-yOmethyl)-4-(2-methoxyethoxy)- 1-
(oxetan-2-
ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid
0
N
0 H
0 N
0
F / __ 0
0 ______________________________________________________ /
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Charge a reaction vessel with (S)-24(54-chloro-16-fluoro-3,7-dioxa-2(2,6)-
pyridina-1(1,3),5(1,2)-dibenzenacyclooctaphane-14-yl)methyl)-4-(2-
methoxyethoxy)-1-
(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (61 mg, 0.087
mmol), 1-
methy1-5-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-y1)pyridin-2(1H)-one (43
mg, 0.174
mmol) and chloro(crotyl)(2-dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-
biphenyl)
palladium(II) (Pd-170, XPhos Pd(crotyl)C1, 6.2 mg, 0.009 mmol). Purge the
vessel with
N2, add DMF (860 iLtL) and aqueous potassium phosphate tribasic (1 M, 260 uL,
0.26
mmol), and stir the mixture at 90 C for 2.5 h. Cool the mixture to RT, add
aqueous citric
acid (5%), filter off the solid and wash it with water and ACN. Purify the
solid by
preparative HPLC [column: XBridge C18, 19 x 150 mm, 5 !dm; mobile phase:
gradient of
30 to 60% ACN in NH4HCO3 (20 mM, pH = 9)] to provide the title compound (9.5
mg,
14%) as a pale brown solid. ES-MS nilz 747 (M+H).
Biological Assays
Human GLP-1 Receptor 11EK293 Cell cAMP Assay
GLP-1 Receptor functional activity is determined using cAMP formation in an
HEK293 clonal cell line expressing human GLP-1R (NCBI accession number
NP 002053) at an expression density of 581 194 (n=6) and 1041 12 (n=5) fmol/mg
protein (determined using [1251]GLp_ 1(7-36)NH2 homologous competition binding
analysis). The hGLP-1R receptor expressing cells are treated with compound (20
point
concentration-response curve in DMSO, 2.75-fold Labcyte Echo direct dilution,
384 well
plate Corning Cat# 3570) in DMEM (Gibco Cat# 31053) supplemented with 1X
GlutaMAXTm (Gibco Cat# 35050), 0.1% bovine casein (Sigma C4765-10ML), 250 tiM
IBMX (3-Isobuty1-1-methylxanthine, Acros Cat # 228420010) and 20 mM HEPES
(Gibco
Cat # 15630) in a 20 1tL assay volume (final DMSO concentration is 0.5%).
After a 30
min incubation at 37 'V, the resulting increase in intracellular cAMP is
quantitatively
determined using the CisBio cAMP Dynamic 2 HTRF Assay Kit (62A1VI4PEJ).
Briefly,
cAlVIP levels within the cell are detected by adding the cAMP-d2 conjugate in
cell lysis
buffer (10 L) followed by the antibody anti-cAMP-Eu3+-Cryptate, also in cell
lysis
buffer (10 p..L). The resulting competitive assay is incubated for at least 60
min at RT,
then detected using a PerkinElmer Envision' instrument with excitation at 320
nm and
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emission at 665 nm and 620 nm. Envision units (emission at 665nm/620nm*10,000)
are
inversely proportional to the amount of cAMP present and are converted to nM
cAMP per
well using a cAMP standard curve. The amount of cAMP generated (nM) in each
well is
converted to a percent of the maximal response observed with human GLP-1(7-
36)NH2.
A relative EC50 value and percent top (Emax) are derived by non-linear
regression analysis
using the percent maximal response vs. the concentration of compound added,
fitted to a
four-parameter logistic equation. The EC50 and Emax data when the compounds of
Examples 1 to 80 are tested in the cAMP assay described above using HEK293
cells
expressing 581 and 104 fmol/mg GLP-1R are shown in Tables 1 and 2,
respectively.
These data indicate that the compounds of Examples 1 to 80 are agonists of the
human
GLP-1 receptor.
Table 1. 11EK293 cell line with 581 fmol/mg expression density of GLP-1R,
intracellular cAMP response, relative EC50 and Emax
Example EC50 nM (SEM, n) Emax % (SEM, n)
1 19.8 (1.6, n = 11) 95 (2, n = 11)
2 8.05 (1.29, n = 8) 98 (3, n = 8)
3 44.1 (3.8, n = 7) 102 (4, n = 7)
4 90.6 (10, n = 7) 99.2 (4.28, n = 7)
5 6.58 (1.44, n = 7) 109 (4.36, n = 7)
6 166 (13.2, n = 8) 97.2 (3.39, n = 8)
7 20.1 (1.59, n = 6) 104 (2.31, n = 6)
8 168 (13.8, n = 6) 90.5 (2.66, n = 6)
9 1.8 (0.162, n= 8) 118 (3.15, n= 8)
10 35.3 (5.55, n = 6) 107 (3.33, n = 6)
11 35 (5.55, n = 6) 92 (3.95, n = 6)
12 20.2 (1.7, n = 8) 81.8 (2.72, n = 8)
13 120 (13.8, n = 6) 88.5 (14.6, n = 7)
14 72.3 (13.7, n = 6) 109 (9.07, n = 6)
6.74 (1.66, n = 7) 105 (3.52, n = 7)
16 1.95 (0.192, n = 7) 125 (5.59, n = 7)
17 10.5 (1.7, n = 4) 121 (4.32, n = 4)
18 50.3 (4.89, n = 7) 115 (1.06, n = 7)
19 55.4 (4.79, n = 8) 103 (2.54, n = 8)
58 (6.74, n = 6) 105 (2.78, n = 6)
21 35.1 (3.83, n = 4) 111 (2.5, n = 4)
22 2.86 (0.448, n = 7) 112 (3.81, n = 7)
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23 69.3 (14.2, n = 9) 106 (4.01, n = 9)
24 2.34 (0.205, n = 5) 116 (1.38, n = 5)
25 317 (n = 1) 98.3 (n = 1)
26 1.66 (0.187, n = 7) 116(5.67, n = 7)
27 50.7 (10.9, n = 5) 107 (4.75, n = 5)
28 98.4 (21.7, n = 5) 104 (3.43, n = 5)
29 262 (26.8, n = 4) 92.4 (5.22, n = 4)
30 35.6 (5.32, n = 5) 98.1 (3.78, n = 5)
31 70.1 (11.7, n = 4) 104 (10.4, n = 4)
32 178 (6.11, n = 3) 98.9(491, n= 3)
33 40.8 (6.25, n = 4) 112 (3.19, n = 4)
34 167 (28.5, n = 4) 110 (2.24, n = 4)
35 95.8 (10.8, n = 3) 112(3.26, n = 3)
36 44 (1.34, n = 4) 108 (7.43, n = 4)
37 179 (28.8, n = 4) 103 (3.15, n = 4)
38 300 (n = 1) 90.1 (n = 1)
39 244 (34.7, n = 4) 99 (1.85, n = 4)
40 42.4 (5.82, n - 4) 110 (6.38, n - 4)
41 184 (11.1, n = 4) 89.1 (3.96, n = 4)
42 187 (22.5, n = 3) 103 (8.87, n = 3)
43 48.6 (3.59, n = 5) 86.9 (3.5, n = 5)
44 4.14 (0.98, n = 5) 112 (6.25, n = 5)
45 2.4 (0.625, n = 4) 122 (3.53, n = 4)
46 7.23 (0.755, n =3) 107 (8.58, n = 3)
47 25.1 (1.86, n = 3) 110(4.79, n = 3)
48 11.2 (1.88, n = 3) 108 (3.83, n = 3)
49 2.33 (0.412, n = 4) 112 (3.83, n = 4)
50 38.1 (4.02, n = 4) 115 (4.45, n = 4)
51 41 (2.51, n = 4) 96.7 (6.62, n = 4)
52 58.4 (16.1, n = 3) 108 (6.21, n = 3)
53 77.8 (13.9, n = 4) 104 (11.3, n = 4)
54 440 (77.8, n = 3) 49.5 (2.94, n = 3)
55 8.94 (1.11, n = 5) 113 (3.06, n = 5)
56 55.8 (13.2, n = 4) 85 (6.71, n = 4)
57 440 (98.7, n = 4) 76.2 (1.11, n = 4)
58 0.633 (0.0868, n = 4) 109 (7.7, n = 4)
59 262 (29.5, n =4) 90.9 (2.1, n = 4)
60 0.938 (0.191, n = 5) 109 (4.24, n = 5)
61 18.1 (3.3, n = 4) 110 (7.36, n = 4)
62 285 (46.7, n = 5) 62.2 (3.62, n = 5)
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63 29.9 (19.1, n = 3) 101 (6.22, n = 3)
64 2.13 (0.188, n = 4) 110 (3.43, n = 4)
65 1.86 (0.365, n = 4) 110 (7.21, n = 4)
66 13.4 (n = 1) 118 (n = 1)
67 2.13 (0.342, n = 4) 117 (6.32, n = 4)
68 6.82 (n = 1) 115 (n = 1)
69 59.8 (3.77, n = 6) 71.6 (2.93, n = 6)
70 3.11 (n = 1) 92.3 (n = 1)
71 138 (9.94, n = 4) 68.4 (8.91, n = 4)
72 92.5 (11.8, n = 6) 106 (4.95, n = 6)
73 21.3 (3.51, n = 3) 106 (4.39, n = 3)
74 155 (n = 1) 92.1 (n = 1)
75 6.33 (1.29, n = 4) 93.9 (3.91, n = 4)
76 102 (33.1, n = 2) 99.9 (1.36, n = 2)
77 193 (18.9, n = 3) 107 (7.26, n = 3)
78 95.8 (10.1, n = 4) 99.8 (4.21, n = 4)
79 369 (20, n = 3) 34.7 (5.13, n = 3)
80 130 (25.3, n - 3) 84.1 (3.1, n - 3)
Table 2. HEK293 cell line with 104 fmol/mg expression density of GLP-1R,
intracellular cAMP response, relative ECso and Emax
Example ECso nM (SEM, Emax (SEM, n)
1 59.7 (9.9, i - 7) 71(3, n - 7)
2 17.5 (3.7, n = 5) 72 (2, n = 5)
3 132 (19, n = 7) 84 (4, n = 7)
4 384 (87.1, n = 7) 78.7 (1.84, n = 7)
5 22.3 (5.33, n = 7) 81.6 (2.84, n = 7)
6 1020 (200, n = 8) 89.1 (2.35, n = 8)
7 96.8 (19.6, n = 6) 92.9 (1.68, n = 6)
8 815 (133, n = 6) 81.5 (3.83, n = 6)
9 5.44 (0.474, n = 8) 83.1 (1.67, n = 8)
147 (22, n = 6) 88.9 (4.9, n = 6)
11 194 (15.5, n = 6) 81 (1.79, n = 6)
12 66.3 (3.11, n = 8) 46.4 (3.52, n = 8)
13 542 (54, n = 6) 74.7 (12.7, n = 7)
14 228 (29, n = 6) 82.5 (2.49, n = 6)
26.4 (3.64, n = 7) 82.8 (2.7, n = 7)
16 4.02 (0.333, n = 5) 86.7 (2.6, n = 5)
17 38.4 (5.56, n = 4) 84.6 (6.33, n = 4)
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18 169 (17.1, n = 7) 94.9 (4.24, n = 7)
19 197 (26.1, n = 8) 83 (2.25, n = 8)
20 226 (33.6, n = 5) 89.9 (3.24, n = 5)
21 133 (25.9, n = 4) 95.6 (3.62, n = 4)
22 10.6 (1.75, n = 7) 80.9 (2.76, n = 7)
23 344 (68.7, n = 9) 83.7 (3.25, n = 9)
24 6.6 (1.03, n = 4) 89.6 (3.8, n = 4)
25 703 (n = 1) 62.1 (n = 1)
26 5.64 (0.655, n = 7) 91.3 (3.5, n = 7)
27 165 (25.4, n = 5) 82.5 (5.93, n = 5)
28 287 (33.3, n= 5) 74.9 (4.14, n = 5)
29 562 (66.1, n = 4) 59.2 (6.36, n = 4)
30 118 (17.3, n = 5) 77.1 (4.75, n = 5)
31 251 (33.7, n = 4) 88.8 (2.84, n = 4)
32 530 (20.8, n = 3) 70.9 (3.4, n = 3)
33 111 (26.1, n = 4) 85 (2.39, n = 4)
34 616 (78.2, n = 4) 82.3 (4.5, n = 4)
35 385 (8.61, n = 3) 88.1 (2.92, n = 3)
36 149 (12.1, n = 4) 80 (3.2, n = 4)
37 506 (57.1, n = 4) 79.7 (3.52, n = 4)
38 613 (n = 1) 56.9 (n = 1)
39 762 (84.7, n = 4) 76.6 (5.29, n = 4)
40 154 (24.3, n = 4) 85.9 (4.29, n = 4)
41 713 (103, n = 4) 60.3 (4.83, n = 4)
42 539 (35.5, n = 3) 66.4 (4.88, n = 3)
43 194 (29.9, n = 5) 72 (5.72, n = 5)
44 16 4 (5.58, n = 5) 85.5 (3.02, n = 5)
45 7.57 (1.86, n = 4) 87.9 (3.62, n = 4)
46 24.6 (5.48, n = 3) 91.7 (2.68, n = 3)
47 116(15.4, n = 3) 108 (2.18, n = 3)
48 31.5 (2.81, n = 3) 86.8 (6.89, n = 3)
49 8.39 (1.12, n = 4) 85.5 (4.12, n = 4)
50 128 (18.8, n = 4) 82.8 (5.92, n = 4)
51 152 (17.9, n = 4) 77.1 (8.25, n = 4)
52 153 (27.5, n = 3) 84.3 (1.76, n = 3)
53 293 (62.7, n = 4) 58 (7.97, n = 4)
54 654 (62.6, n = 3) 18.3 (5, n = 3)
55 22.8 (2.26, n = 5) 84.8 (5.59, n = 5)
56 131 (27.6, n = 4) 41.7 (3.43, n = 4)
57 1030 (212, n = 4) 42.1 (2.24, n = 4)
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58 1.62 (0.195, n = 4) 90 (6.63, n = 4)
59 515 (38.2, n = 4) 48.4 (6.45, n = 4)
60 2.99 (0.356, n = 5) 87.9 (4.63, n = 5)
61 79.1 (10.5, n = 4) 93.2 (9.3, n = 4)
62 737 (41.9, n = 5) 32.9 (3.9, n = 5)
63 112 (72.1, n = 3) 81.1 (4.02, n = 3)
64 5.53 (0.884, n = 4) 87.3 (2.6, n = 4)
65 4.62 (0.69, n = 4) 98.2 (2.37, n = 4)
66 54.5 (n = 1) 82.5 (n = 1)
67 6.88 (0.996, n = 4) 91.6 (2.82, n = 4)
68 8.72 (n = 1) 98.5 (n = 1)
69 103 (14.3, n = 5) 26.6 (3.01, n = 5)
70 12.6 (n = 1) 95 (n = 1)
71 598 (61.9, n = 4) 52.1 (9.13, n = 4)
72 348 (33.8, n = 6) 90.4 (4.48, n = 6)
73 68.5 (2.21, n = 3) 84.1 (3.4, n = 3)
74 533 (n = 1) 73.3 (n = 1)
75 17.4 (3.1, n = 4) 71.6 (8.14, n = 4)
76 359 (113, n = 2) 78.2 (7.88, n = 2)
77 724 (42.4, n = 3) 83 (5.01, n = 3)
78 248 (12.6, n = 4) 74.1 (3.39, n = 4)
79 648 (47.3, n = 3) 9.53 (2.66, n = 3)
80 303 (26, n = 3) 56 (5.21, n = 3)
EC50, nM = geometric mean followed by SEM (delta method) with the number of
observations in
parentheses.
Ema, % = arithmetic mean followed by SEM with the number of observations in
parentheses for the
percent of maximal response to GLP-1(7-36)NH2 at hGLP-1R
GLP-1R CHO Cell P-Arrestin Recruitment Assay
Activated G-protein coupled receptors can interact with the P-arrestin family
of
signaling proteins. The potency of compounds for GLP-1R induced arrestin
recruitment is
determined using the PathHunter Enzyme Fragment Complementation approach
substantially as described (von Degenfeld et cii., FASEB J., 2007 (14):3819-26
and
Hamdouchi et al., J. Med Chem., 2016 59(24):10891-10916). CHO-Kl cells
expressing
Pro-Link-tagged Human GLP-1R and enzyme-acceptor-tagged 13-arrestin-2 may be
obtained from DiscoveRx and prepared as assay-ready frozen cells. Test
compounds are
solubilized in DMSO and serial dilutions are performed using the Echo acoustic
dispenser
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(LabCyte). Assay media is the PathHunter Cell Assay Buffer (DiscoveRx)
containing
0.1% w/v hydrolyzed Casein (Sigma). 100 nL of test compound solution is
dispensed into
pL of assay media in a 384 well plate and then 10 pL of cells in assay media
are added
to give 5000 cells per well. Plates are incubated for 90 min in a 37 C/5% CO2
incubator
5 and 10 pL of PathHunter detection reagent is added (DiscoveRx) and plates
are incubated
at RT for 60 min. Luminescence signal is measured. Compound concentration-
response
curves fit to a four-parameter logistic model to calculate potency as an EC50
and percent
top (Emax). Data normalization to % stimulation is performed using DMSO and
GLP-1(7-
36) as minimum and maximum controls (Campbell et al, Assay Guidance Manual
2017).
10 The potency of a sample compound to stimulate GLP-1R induced p-arrestin
recruitment
is reported in Table 3.
Table 3. hGLP1R induced P-Arrestin-2 recruitment relative EC50 and Emax
Example EC50 nM (SEM, n) Emax % (SEM, n)
1 213 (36.7, n = 9) 42.9 (1.85, n = 9)
2 133 (6.65, n = 7) 52.7 (1.78, n = 7)
3 320 (29.4, n = 7) 34 (2.44, n = 7)
12 >49500 (n = 1) 9.47 (1.44, n = 7)
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Representative Drawing