INTERMEDIATES FOR THE PREPARATION OF ANALOGS OF HALICHONDRIN B

INTERMEDIAIRES POUR LA PREPARATION D'ANALOGUES D'HALICHONDRINE B

English Abstract

The present invention provides macrocyclic compounds, synthesis of the same
and intermediates
thereto. Such compounds, and compositions thereof, are useful for treating or
preventing
proliferative disorders Formula (F-4).

French Abstract

La présente invention a trait à des composés macrocycliques, la synthèse de ceux-ci et leurs intermédiaires. De tels composés, et leurs compositions, servent à traiter ou éviter les troubles prolifératifs de formule F4.

Claims

THE EMBODIMENTS OF THE INVENTION FOR WHICH AN EXCLUSIVE PROPERTY OR
PRIVILEGE IS CLAIMED AR B DEFINED AS FOLLOWS:
1. A compound of formula A, B, or C:
Image
wherein:
~ designates a single or double bond;
n is 1, 2, or 3;
W is CH-A or C(O);
A is a C1-6 aliphatic group, wherein A is optionally substituted with one or
more Q1 groups;
each Q1 is independently selected from the group consisting of cyano, halo,
azido, oxo, OR, SR,
SO2R, OSO2R, N(R)2, NR(CO)R, NR(CO)(CO)R, NR(CO)N(R)2, NR(CO)OR, (CO)OR,
O(CO)R, (CO)N(R)2, O(CO)N(R)2, and OPG1, wherein:
optionally two Q1 are on A and are taken together to form a 3-8 membered
saturated,
partially unsaturated, or aryl ring having 0-4 heteroatoms independently
selected from
the group consisting of nitrogen, oxygen, and sulfur;
each R is hydrogen or an optionally substituted group selected from the group
consisting of C1-6
aliphatic, a 5-10 membered saturated, partially unsaturated or aryl
carbocyclic ring, and a 4-
membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms

independently selected from the group consisting of nitrogen, oxygen, and
sulfur, wherein:
when Q1 is independently selected from the group consisting of N(R)2,
NR(CO)N(R)2,
(CO)N(R)2, and O(CO)N(R)2, two R groups on the same nitrogen atom are
optionally
taken together with said nitrogen atom to form a 3-8 membered saturated,
partially
unsaturated, or aryl ring having 1-4 heteroatoms independently selected from
the
group consisting of nitrogen, oxygen, and sulfur; and
wherein X B; is O, and R B is TMS; X B is H and OH, and R B is TMS; or X B is
H and OAc, and R B
is Ac; and R1 is ¨CH2CO2R2, wherein R2 is alkyl,
92

wherein PG1, PG5, and PG6 are hydroxyl protecting groups, which taken with the
oxygen
atom to which each is bound, are independently selected from the group
consisting of an ester, a
silyl ether, an alkyl ether, an arylalkyl ether, and an alkoxyalkyl ether, or
PG5 and PG6 are taken
together to form a cyclic diol protecting group, which taken together with the
oxygen atoms to
which it is bound, is selected from the group consisting of a cyclic acetal, a
cyclic ketal, a
silylene derivative, a cyclic carbonate, and a cyclic boronate.
2. The compound of claim 1 having the formula:
Image
3. The compound of claim 2 having the formula:
Image
4. The compound of claim 1 having the formula:
Image
5. The compound of claim 1 having the formula:
Image
93


6. The compound of claim 1 having the formula:
Image
7. The compound of claim 1 having the formula:
Image
8. The compound of claim 1, wherein the compound is of formula C, and R2 is
methyl.
94

Description

CA 02822994 2013-08-01
INTERMEDIATES FOR THE PREPARATION OF ANALOGS OF HALICHONDRIN B
[0001]
TECHNICAL FIELD OF INVENTION
[0002] The present invention relates to compounds useful as intermediates
in the
synthesis of pharmaceutically active macrolide compounds.
BACKGROUND OF THE INVENTION
[0003] The invention relates to pharmaceutically active macrolides,
synthesis thereof and
intermediates thereto. Halichondrin B is a potent anticancer agent originally
isolated from
the marine sponge Halichondria okadai, and subsequently found in Axinella sp.,
Phakellia
carteri, and Lissondendryx sp. A total synthesis of Halichondrin B was
published in 1992
(Aicher, T. D. et al., J. Am_ Chem. Soc. 114:3162-3164). Halichondrin B has
demonstrated
in vitro inhibition of tubulin polymerization, microtubule assembly, betas-
tubulin
crosslinking, GTP and vinblastine binding to tubulin, and tubulin-dependent
GTP hydrolysis
and has shown in vitro and in vivo anti-cancer properties. Accordingly, there
is a need to
develop synthetic methods for preparing analogs of Halichondrin B useful as
anti-cancer
agents.
SUMMARY OF THE INVENTION
[0004] As described herein, the present invention provides methods for
preparing analogs
of Halichondrin B having pharmaceutical activity, such as anticancer or
antimitotic (mitosis-
blocking) activity. These compounds include a compound of formula B-1939:
1

CA 02822994 2013-08-01
Me0,
HO
ss= = 0 0
0
=0
"==
I 0
B-1939.
[0005] These compounds are useful for treating cancer and other
proliferative disorders
including, but not limited to, melanoma, fibrosarcoma, leukemia, colon
carcinoma, ovarian
carcinoma, breast carcinoma, osteosarcoma, prostate carcinoma, and lung
carcinoma. The
present invention also provides synthetic intermediates useful for preparing
said analogs of
Halichondrin B.
2

CA 02822994 2013-08-01
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION
[0006] The methods and intermediates of the present invention are useful
for preparing
various analogs of Halichondrin B as described in, e.g. United States Patent
6,365,759 and
United States Patent 6,469,182.
These Halichondrin B analogs are prepared generally by the assembly of three
fragments F-1,
F-2, and F-3, as shown by Scheme I below:
Scheme I
R1 SO2Ph
______________________________________________________________________ . B-
1939
R2
(NOPG2 F-1
OPG1 , X
LG '
OPG4 _________________________________________________
0
LG2
OPG3 R3
0
OPG4r..nrs4
F_3
2a

CA 02822994 2013-08-01
I. Fragment F-1
[0007] According to one embodiment, the present invention provides a
compound F-1:
R1 so2Rh
R2
0
rNoRG2
oRG1
F-1
wherein:
each of PG' and PG2 is independently hydrogen or a suitable hydroxyl
protecting group;
RI is R or OR;
R2 is CHO or ¨CH=CH2; and
each R is independently hydrogen, Ci_4 haloaliphatic, benzyl, or CM aliphatic,
provided that
when RI is OMe then PG' and PG2 do not form an acetonide group.
[0008] In certain embodiments, RI is OR. In other embodiments, RI is OR
wherein R is
hydrogen, methyl, or benzyl.
[0009] In certain embodiments, PG' and PG2 are hydrogen. In other embodiments,
one
of PG' and PG2 is hydrogen.
[0010] Suitable hydroxyl protecting groups are well known in the art and
include those
described in detail in Protecting Groups in Organic Synthesis, T. W. Greene
and P. G. M.
Wuts, 3rd edition, John Wiley & Sons, 1999.
In certain embodiments, each of PG' and PG2, taken with the oxygen atom to
which it is
bound, is independently selected from esters, ethers, silyl ethers, alkyl
ethers, arylallcyl ethers,
and alkoxyalkyl ethers. Examples of such esters include formates, acetates,
carbonates, and
gillfonates. Specific examples include formate, benzoyl formate,
chloroacetate,
trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-
chlorophenoxyacetate, 3-
phenylpropionate, 4-oxopentanoate, 4,4-(ethylenedithio)pentanoate, pivaloate
(trimethylacetyl), crotonate, 4-methoxy-crotonate, benzoate, p-benzylbenzoate,
2,4,6-
trimethylbenzoate, or carbonates such as methyl, 9-fluorenylmethyl, ethyl,
2,2,2-
trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl,
allyl, and p-nitrobenzyl.
Examples of such silyl ethers include trimethylsilyl, triethylsilyl, t-
butyldimethylsilyl, t-
butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl ethers. Alkyl
ethers include
methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, ally',
and
3

CA 02822994 2013-08-01
allyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers include acetals
such as
methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl, benzyloxymethyl,
beta-
(trimethylsilyl)ethoxymethyl, and tetrahydropyranyl ethers. Examples of
arylalkyl ethers
include benzyl, p-methoxybenzyl (IVIPM), 3,4-dimethoxybenzyl, o-nitrobenzyl, p-

nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2- and 4-
picolyl.
[0011] In certain embodiments, one or both of the PG1 and PG2 moieties of F-
1 are silyl
ethers or arylallcyl ethers. In yet other embodiments, one or both of the PG'
and PG2
moieties of F-1 are t-butyldimethylsilyl or benzoyl. In still other
embodiments, both of the
PG' and PG2 moieties of F-1 are t-butyldimethylsilyl.
[0012] According to an alternate embodiment, PG' and PG2 are taken together,
with the
oxygen atoms to which they are bound, to form a diol protecting group, such as
a cyclic
acetal or ketal. Such groups include methylene, ethylidene, benzylidene,
isopropylidene,
cyclohexylidene, and cyclopentylidene, a silylene derivative such as di-t-
butylsilylene and a
1,1,3,3-tetraisopropyldisiloxanylidene derivative, a cyclic carbonate, and a
cyclic boronate.
Methods of adding and removing such hydroxyl protecting groups, and additional
protecting
groups, are well-known in the art and available, for example, in P. J.
Kocienski, Protecting
Groups, Thieme, 1994, and in T. W. Greene and P. G. M. Wuts, Protective Groups
in
Organic Synthesis, 3`d edition, John Wiley & Sons, 1999. According to another
embodiment, PG' and PG2 are taken together to form an acetonide group.
[0013] According to one embodiment, R2 is CHO.
[0014] According to another embodiment, R2 is --CH=CH2-
[0015] In certain embodiments, the present invention provides a compound of
formula F-
1 having the stereochemistry depicted in compound F-1':
RI so2Ph
õ _________________________________ (
opG,
F-1'
wherein each variable is as defined above and described in classes and
subclasses above and
herein.
[0016] In certain embodiments, the following compounds F-la and F-lb are
provided:
4

CA 02822994 2013-08-01
Me0, SO2Ph HO., SO2Ph
ss,4o
r0TBS
OTBS OH
F-la F-1 b
wherein "TBS" refers to t-butyldimethylsilyl.
[0017] Details of the syntheses of F-la and F-lb are set forth in the
Examples infra.
2. Fragment F-2
[0018] According to another embodiment, the present invention provides a
compound
F-2:
LG1 X
)\/\
=
0
F-2
wherein:
each -- is independently a single or double bond, provided that both groups
are not
simultaneously a double bond;
LGI is a suitable leaving group;
X is halogen or ¨0SOARY);
RY is C1-6 aliphatic or a 5-7 membered saturated, partially unsaturated, or
fully unsaturated
ring, wherein RY is optionally substituted with up to 3 groups selected from
halogen, R,
NO2, CN, OR, SR, or N(R)2;
each R is independently hydrogen, C14 haloaliphatic, or Ci4 aliphatic; and
PG3 is a suitable hydroxyl protecting group.
[0019] As used herein, a suitable leaving group is a chemical moiety that
is readily
displaced by a desired incoming chemical moiety. Suitable leaving groups are
well known
in the art, e.g., see, "Advanced Organic Chemistry," Jerry March, 4th Ed., pp.
351-357, John
Wiley and Sons, N.Y. (1992). Such leaving groups include, but are not limited
to, halogen,
alkoxy, sulphonyloxy, optionally substituted alkylsulphonyloxy, optionally
substituted
alkenylsulfonyloxy, optionally substituted arylsu I fonyloxy, and diazonium
moieties.

CA 02822994 2013-08-01
Examples of suitable leaving groups include chloro, iodo, bromo, fluoro,
methanesulfonyloxy (mesyloxy), tosyloxy, triflate, nitro-phenylsulfonyloxy
(nosyloxy), and
bromo-phenylsulfonyloxy (brosyloxy). In certain embodiments, the LG1 moiety of
F-2 is
sulphonyloxy, optionally substituted alkylsulphonyloxy, optionally substituted

alkenylsulfonyloxy, or optionally substituted arylsulfonyloxy. In other
embodiments, the
LG1 moiety of F-2 is optionally substituted alkylsulphonyloxy. In yet other
embodiments,
the LGI moiety of F-2 is mesyloxy or tosyloxy.
[0020] In certain embodiments, the X moiety of F-2 is halogen. In other
embodiments,
the X moiety of F-2 is sulphonyloxy, optionally substituted alkylsulphonyloxy,
optionally
substituted alkenylsulfonyloxy, or optionally substituted arylsulfonyloxy. In
still other
embodiments, the X moiety of F-2 is triflate.
[0021] In certain embodiments, the PG3 moiety of F-2, taken with the oxygen
atom to
which it is bound, is a silyl ether. In other embodiments, the PG3 moiety of F-
2, taken with
the oxygen atom to which it is bound, is an ester group_ According to one
aspect of the
present invention, the PG3 moiety of F-2 is t-butyldimethylsilyl. According to
another
aspect of the present invention, the PG3 moiety of F-2 is pivaloyl or benzoyl.
[0022] In certain embodiments, the present invention provides a compound of
formula F-
2 having the stereochemistry depicted in formula F-2':
LG, X
0
F-2'
wherein each variable is as defined above and described in classes and
subclasses above and
herein.
[0023] In certain embodiments, a compound F-2a or F-2b is provided:
Ms0 Ts0
rC; 0
F-2a F-2b
6

CA 02822994 2013-08-01
wherein "Ms0" refers to mesylate, "Tf0" refers to triflate, "OPv" refers to
pivaloate, "OBz"
refers to benzoate, and "Ts0" refers to tosylate.
[0024] In other embodiments, the present invention provides a compound of
formula F-
2b wherein said compound is crystalline. According to another embodiment, a
compound of
formula F-2b is provided wherein said compound is crystallized from an alkane
solvent. In
certain embodiments, crystalline F-2b is provided wherein said compound is
crystallized
from pentane or heptane. In other embodiments, crystalline F-2b is provided
wherein said
compound is crystallized at about 0 C.
[0025] Compounds of formula F-2 are prepared generally from intermediates F-
2d and
F-2e as shown in Scheme A below.
Scheme A
Alk OPG5
N
R PG50 PG50 0
0
F-2d Alk a"--""-Alk F-2
OH
LG4 -o OPG3
OPG3
OPG
F-2e
[0026] Accordingly, another aspect of the present invention provides a
compound of
formula F-2d:
Alk OPG5
0,N
R'
I 0
F-2d
wherein:
R' is ¨CH=CH2 or -C(0)H;
Alk is a CIA straight or branched aliphatic group; and
PG5 is a suitable hydroxyl protecting group.
[0027] Suitable hydroxyl protecting group PG5 is as described and defined
for the PG3
moiety of compound F-2, supra. In certain embodiments, PG5, taken with the
oxygen atom
to which it is bound, is a silyl ether. In other embodiments, PG5 is t-
butyldimethylsilyl.
[0028] According to one embodiment, the Alk moiety of compound F-2d is methyl.
7

CA 02822994 2013-08-01
[0029] In certain embodiments, a compound of formula F-2d' is provided:
Alk OPG5
,N
0 R'
I 0
F-2d'
[0030] Yet another aspect of the present invention provides a compound of
formula F-2e:
R"
Br OPG3
F-2e
wherein:
R" is OH, 0PG3, or LG4;
LG4 is a suitable leaving group; and
each PG3 is independently a suitable hydroxyl protecting group, provided that
R" is other
than OMs when PG3 is t-butyldiphenylsilyl.
[0031] One of ordinary skill in the art would recognize that the R" moiety
of compound
F-2e may be transformed from OH to a protected hydroxyl group, OPG3, or,
alternatively,
directly to LG4. Such transformations are known to one skilled in the art and
include,
among others, those described herein. In certain embodiments, R" is OH or LG4.
The LG4
leaving group of formula F-2e is as described and defined for the LG1 moiety
of compound
F-2, supra. In certain embodiments, LG4 is tosyloxy or mesyloxy.
[0032] The PG3 moiety of compound F-2e is as defined and described for the PG3
moiety
of compound F-2, supra. In certain embodiments, PG3, taken with the oxygen
atom to
which it is bound, is a silyl ether. In other embodiments, PG3 is t-
butyldiphenylsilyl.
[0033] Still another aspect of the present invention provides a compound F-
2f:
0
(31!/
PG-5 0 '===
AIk
OPG3
F-2f
wherein Alk, PG3 and PG3 are as defined generally and in classes and
subclasses described
above and herein. Compounds of formula F-2f are used to prepare compounds of
formula
F-2 by methods described herein and those known in the art.
8

CA 02822994 2013-08-01
[0034] Details of the synthesis of F-2a are set forth in the Examples
infra.
[0035] Alternatively, compounds of formula F-2 are prepared from D-quinic
acid as
shown by Scheme II below. Details of the preparation of compounds of formula F-
2 are set
forth in the Examples infra.
9

CA 02822994 2013-08-01
Scheme II
TMS
HO2C 1. Cyclohexanone1) Fi
OH H+ , X\\___ )=,..-
2. TMSCI ______________________ RO'' )0 acid cat. RO'V i
õ
:
3. DIBAL
OH
4. Ac20 2, X=0, R=H 6a, R= Ac, R1 =
CH2CO2R2
1, D-quinic acid 3, X=0, R=TMS R2= alkyl
4, X=1-1, OH, R=TMS
6b, R= Ac, R1 =I-I
5, X=H, OAc, R=Ac
from 6a H
Na0Me 0 õH H
1
, HCI
,,,,....40?5:H . BrCOC(Me)20Ac
2 Na0Me
1.9-BBN
from 6b 4 ,,c,
Ro2c
Me0H 2. NaBH4, Me0H
ROC 0
2. [0] 7, R= alkyl b----b 8, R= alkyl ,
OH 3. TBDPSCI,
NEt3
3. HWE or Wittig
4. Na0Me
H
H .H 1. 03; NaBH4
LDA
R ------.- TBDPSO 0 14 = ,'OH 2. Nal(/'
0 ,=, 3. Ph3PCHCO2Me
9, R=CO2R1 "0 12
R1= alkyf or H
10, R=CH2OH
11, R=CH2OTBDPS
H
0 1.
Reduction
fl_O 2. TrCI,
NEt3
TBDPSO X
H2, Pd/C
0
OH TBDPSO 0"----*\----0O2Me 3. TBAF, THF
OH
13, X=OH, CH2OH
16
14, X=0
15, X=CHCO2Me
0
0 1. Zn, Et0H
1. NIS, PPh3
. .
R'''-'-'"'"c-r-----)--N'
\ 2. LDA; Mel
2. KCN, Et0H/H20 ''R' OTr
OR' OR"
17, R=TBDPS, R'=R"=H 20, R=R'=I
18, R=TBDPS, R'=H, R"=Tr 21, R=CN, R'=I
19, R=R'=H, R"=Tr
Y X
0
0 r OR
õ. '
I. HN(OMe)Me TBSO OMe 1. MeMgBr (1"---1:"Me
AlMe3 Me
2. LDA, NPTf ==)- OR,
2. TBSCI
10 0Tr 3' HCI' IPA
22, R=H 24 25, X=0,Y=Me, R=TBS, R'=Tr
26, X=CH2, Y=0Tf, R=TBS, R'=Tr
23, R=Me 27, X=CH2, Y=0-11, R=R'=H
+
[0036] Yet another method for preparing compounds of formula F-2 from D-
quinic acid
provides an alternative route from intermediate 12 to intermediate 17 as shown
in Scheme
III below.

CA 02822994 2013-08-01
Scheme III
0 epoxide 0 õH oxidation
Rx 0 114 ','OH formation Rx
W."0H
_
12
0 -NHNH2
0
Rx 0 11110 0 so
_
-
CH2Cl2, AcOH CHO
NaBH4 EtMgBr or BuLi OH
¨ /
Et0H CH20
Rx 0 Rx 0
OH OH
H2, Pd/C
Rx
Et0H
OH ON
17
[0037] Scheme III above shows an alternate method for preparing
intermediate 17 from
intermediate 12 via Eschenmoser-Tanabe Fragmentation, wherein each Rx is
independently
OPW or CN wherein PG' is a suitable hydroxyl protecting group as described
herein.
Intermediate 17 is then used to prepare compounds of formula F-2 according to
Scheme II
above.
[0038] Still another method for preparing compounds of formula F-2 from D-
quinic acid
provides an alternative route from intermediate 9 to intermediate 17 as shown
in Scheme IV
below.
11

CA 02822994 2013-08-01
Scheme IV
H
H H 0 ,H
0.H LDA ,s
,
Deprotection¨ HO2C
PGYO2C
0 VP- ______________________ HO2C 0 Imio_ 0 4.,.OH
""b '''d
9
H H
0
sõH oxidation
epoxide HO2C HO2C
0 Nio "'OH _____________________________ _ 0 IWO 0
formation -
_ =
0' b's
H H
110 NHNH2 0 --'-',_.---0
reduction ----
I \
--
00 HO2Cr:-----)(1 ¨
¨ CHO
CH2Cl2, AcOH 'OH
H
Hr",..--0 OH
EtMgBr or BuLi
Hydroxyl protection 0
¨
CH20 PGx00
OH'
OH
H
H2, Pd/C
Et0H PGx00--------1¨\--\
OH OH
17
wherein PG Y is a suitable carboxyl protecting group, as described herein, and
each PG" is
independently a suitable hydroxyl protecting group as described herein.
[0039] Yet
another method for preparing intermediates useful for preparing compounds
of formula F-2 from D-quinic acid is shown in Scheme V below.
12

CA 02822994 2013-08-01
Scheme V
/=,, 0 , 0
Me02C, LAH
HOCCV:i1:?),
'OPG6 THF 0 C ''OPG6
,
A-1 A-2
OPG5 bPG5
KCN
LG 5 " '0 PG 6 0. = "OPG6
Et0H, H20
0PG5 80 *C OPG5
A-3 A-4
/'"
O.,, 0
1. Deprotection 0
2. recrystallization NC
ER-817664 OH
[0040] In Scheme V above, intermediate 7 (from Scheme II), wherein R is a
methyl ester,
is used to prepare ER-817664 as a crystalline intermediate. As depicted in
Scheme V above,
each PG5 and PG6 is independently a suitable hydroxyl protecting group. In
certain
embodiments, PG5 and PG6 are taken together to form a cyclic diol protecting
group. In
other embodiments, PG5 and PG6 are taken together to form a cyclohexylidene
protecting
group. As depicted in Scheme V above, LG5 is a suitable leaving group. Such
suitable
leaving groups are well known in the art and include those described herein.
In certain
embodiments, LG5 is mesyloxy or tosyloxy.
[0041] According to another embodiment, the present invention provides a
compound of
formula A:
0
w,r7- via 0,G6
0
OPG5
A
wherein:
designates a single or double bond;
n is 1, 2, or 3;
each of PG5 and PG6 is independently a suitable hydroxyl protecting group;
W is CH-A or C(0);
13

CA 02822994 2013-08-01
A is a C,6 aliphatic group, wherein A is optionally substituted with one or
more Q1 groups;
each Q1 is independently selected from cyano, halo, azido, oxo, OR, SR, SO2R,
OSO2R,
N(R)2, NR(CO)R, NR(C0)(CO)R, NR(CO)N(R)2, NR(C0)0R, (C0)0R, 0(CO)R,
(CO)N(R)2, 0(CO)N(R)2, or OPG1, wherein PG' is a suitable hydroxyl protecting
group,
and wherein:
two Q1 on A are optionally taken together to form a 3-8 membered saturated,
partially
unsaturated, or aryl ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur; and
each R is independently selected from hydrogen or an optionally substituted
group selected
from Ci_6 aliphatic, a 5-10 membered saturated, partially unsaturated or aryl
carbocyclic
ring, or a 4-10 membered saturated, partially unsaturated or aryl ring having
0-4
heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein:
two R groups on the same nitrogen atom are optionally taken together with said

nitrogen atom to form a 3-8 membered saturated, partially unsaturated, or aryl
ring
having 1-4 heteroatoms independently selected from nitrogen, oxygen, or
sulfur.
[0042] In certain embodiments, the present invention provides a compound of
formula A
having the stereochemistry as depicted in formula A':
0
,OPG6
0' s ______________________________
'OPG5
A'
wherein each variable is as defined above and described in classes and
subclasses above and
herein.
[0043] In certain embodiments, the present invention provides a compound of
formula A'
wherein W is C(0) and said compound is of formula A'-1:
0
r
..,0pG6
0
0,
,0PG5
A'-1
14

CA 02822994 2013-08-01
wherein each variable is as defined above and described in classes and
subclasses above and
herein_
[0044] As defined generally above, the A group of formulae A and A' is a
C1_6 aliphatic
group, wherein A is optionally substituted with Q1. In certain embodiments,
the A group of
formulae A and A' is a C2_5 aliphatic group, wherein A is substituted with one
or more Q1
groups.
[0045] As defined generally above, each Q1 group of formulae A and A' is
independently
selected from cyano, halo, azido, oxo, OR, SR, SO2R, OSO2R, N(R)2, NR(CO)R,
NR(C0)(CO)R, NR(CO)N(R)2, NR(C0)0R, (C0)0R, 0(CO)R, (CO)N(R)2, 0(CO)N(R)2,
or OPGI, wherein PG' is a suitable hydroxyl protecting group. In certain
embodiments, each
Q1 group of formulae A and A' is independently selected from cyano, halo,
azido, oxo,
N(R)2, OR, SR, SO2R, or OSO2R. In other embodiments, each Q1 group of formulae
A and
A' is independently selected from cyano, halo, azido, oxo, OR, SR, SO2R,
OSO2R, N(R)2,
NR(CO)R, NR(CO)R, and 0(CO)N(R)2. In still other embodiments, exemplary Q1
groups
include NH(C0)(C0)-(heterocyclic radical or heteroaryl), 0S02-(aryl or
substituted aryl),
0(CO)NH-(aryl or substituted aryl), aminoalkyl, hydroxyalkyl, NH(C0)(C0)-(aryl
or
substituted aryl), N-1-1(C0)(alkyl)(heteroaryl or heterocyclic radical),
0(substituted or
unsubstituted alkyl)(substituted or unsubstituted aryl), and
NH(C0)(alkyl)(aryl or
substituted aryl).
[0046] In certain embodiments, the A group of formulae A and A' has one of
the
following characteristics:
(1) A has at least one substituent selected from hydroxyl, amino, azido, halo,
and oxo;
(2) A is a C1.6 alkyl group having at least one substituent selected from
hydroxyl,
amino, and azido;
(3) A has at least two substituents independently selected from hydroxyl,
amino, and
azido;
(4) A has at least two substituents independently selected from hydroxyl and
amino;
(5) A has at least one hydroxyl substituent and at least one amino
substituent;
(6) A has at least two hydroxyl substituents;
(7) A is a C2_4 aliphatic group that is substituted;
(8) A is a C3 aliphatic group that is substituted;

CA 02822994 2013-08-01
(9) A has an (S)-hydroxyl alpha to the carbon atom linking A to the ring
containing G
or an (R)-hydroxyl; and
(10) A is a Ci_6 saturated aliphatic group having at least one substituent
selected from
hydroxyl and cyano.
[0047] The term "(S)-hydroxyl" means that the configuration of the carbon
atom having
the hydroxyl group is (S). Embodiments of the invention also include compounds
wherein
A is substituted at least once on each carbon atom: (1) alpha and gamma, (2)
beta and
gamma, or (3) alpha and beta to the carbon atom to which A is attached. Each
of the alpha,
beta, and gamma carbon atoms are independently in the (R) or (S)
configuration. In certain
embodiments, the invention provides said compound wherein A is substituted at
least once
on each carbon atom alpha and beta to the carbon atom to which A is attached.
[0048] Exemplary A groups of formulae A and A' include 2,3-dihydroxypropyl, 2-
hydroxyethyl, 3-hydroxy-4-perfluorobutyl, 2,4,5-trihydroxypentyl, 3-amino-2-
hydroxypropyl, 1,2-dihydroxyethyl, 2,3-dihyroxy-4-perflurobutyl, 3-cyano-2-
hydroxypropyl, 2-amino-l-hydroxy ethyl, 3-azido-2-hydroxypropyl, 3,3-difluoro-
2,4-
dihydroxybutyl, 2,4-dihydroxybutyl, 2-hydroxy-2-(p-fluoropheny1)-ethyl, -
CH2(C0)(substituted or unsubstituted aryl), -CH2(C0)(allcyl or substituted
alkyl, such as
haloalkyl or hydroxyalkyl) and 3,3-difluoro-2-hydroxypent-4-enyl.
[0049] In certain embodiments, the A group of either of formulae A and A'
is 3-amino-2-
hydroxypropyl.
[0050] According to one aspect, the present invention provides a compound
of either of
formulae A and A', wherein Q1 is OPG1, wherein PG' is a suitable hydroxyl
protecting
group. Suitable hydroxyl protecting groups are well known in the art and
include those
described in detail in Protecting Groups in Organic Synthesis, T. W. Greene
and P. G. M.
Wuts, 31-d edition, John Wiley & Sons, 1999.
In certain embodiments, the PG' moiety of either of formulae A and A', taken
with the
oxygen atom to which it is bound, is selected from esters, ethers, silyl
ethers, alkyl ethers,
arylalkyl ethers, and alkoxyalkyl ethers. Examples of such esters include
formates, acetates,
carbonates, and sulfonates. Specific examples include formate, benzoyl
formate,
chloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-
chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate, 4,4-
(ethylenedithio)pentanoate,
pivaloate (trimethylacetyl), crotonate, 4-methoxy-crotonate, benzoate, p-
benzylbenzoate,
16

CA 02822994 2013-08-01
2,4,6-trimethylbenzoate, carbonates such as methyl, 9-fluorenylmethyl, ethyl,
2,2,2-
trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl,
allyl, and p-nitrobenzyl.
Examples of such sit)/ ethers include trimethylsilyl, triethylsilyl, t-
butyldimethylsilyl, t-
butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl ethers. Alkyl
ethers include
methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, ally!,
and
allyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers include acetals
such as
methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl, benzyloxymethyl,
beta-
(trimethylsilypethoxymethyl, and tetrahydropyranyl ethers. Examples of
arylalkyl ethers
include benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, o-nitrobenzyl, p-
nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2- and 4-
picolyl.
[0051] In certain embodiments, the PG1 moiety of either of formulae A and
A', taken
with the oxygen atom to which it is bound, is a silyl ether or arylalkyl
ether. In yet other
embodiments, the PG1 moiety of either of formulae A and A' is t-
butyldimethylsilyl or
benzoyl. In still other embodiments, the PG1 moiety of either of formulae A
and A' is t-
butyldimethylsily1("TBS").
[0052] As defined generally above, two Q1 on A are optionally taken
together to form a
3-8 membered saturated, partially unsaturated, or aryl ring having 0-4
heteroatoms
independently selected from nitrogen, oxygen, or sulfur. In certain
embodiments, two Q1 on
A are taken together to form an epoxide ring.
[0053] In certain embodiments, the PG5 and PG6 groups of formula A and A'
are
independently selected from those suitable protecting groups described above
for the PG1
group of formula A and A'. In other embodiments, the PG5 and PG6 groups of
formula A
and A' are taken together to form a cyclic diol protecting group. Such diol
protecting groups
are well known in the art and include those described by Greene and include
cyclohexylidene and benzylidene diol protecting groups.
[0054] In certain embodiments, the present invention provides a method for
preparing
compounds of formula 11-2 according to Schemes V-a, V-b, and V-c below:
17

CA 02822994 2013-08-01
Scheme V-a
HO2C
H0"10OH 0 lh .1-1 0 _IA
/. 0 0.--
cyc_oexanone TMSCI
õ ...0 .-0
_ OH

H2SO4 imid. THE TMSCYs \'C )0
OH 0 '0
D-quinic acid ER-108423 ER-812929
o ,c.,,,
0 ,H Ac0 TMS
H
HO--..y5 Me02C,,
DIBALH Ac20, AcOH õ ' ...0
Aca _______________________________________
Tolune TMSOssµ )0 BF30Et2,
TFAA
."0 CH3CN, it
ER-812930 ER-812933
H 0
Me02C, õ,,(y fli
Na0Me, Me0H Me02C
Ac0A1 i AO THF
-"0-0ER-812934 ER-812935
H H
0 0
õH MsCI,Et3N KCN, E10H
LAH, THE
' HO::.""0 Ms0 0
õ, 10
0 oti
ER-816961 ER-818937
H H
0 0
,.H
HCI, Me0H
. NC-'-'j411:)."0 NC 'OH
recrystallization
--0-10-OH
ER-818950 ER-817664
crystalline single isomer
[0055] In other embodiments, the present invention provides crystalline ER-
817664.
18

CA 02822994 2013-08-01
Scheme V-b
H 0 H
0 0
H
Br )ç0
õH
,.= ,
DBU
NC 0 1111=."OH ________ NC 0 4."0Ac
toluene
CI-13CN, H20
ER-817664 OH
ER-818635 Br
H
H 1.03, Me0H /\--0
0 õsH 2. NaBH4, õ.H (Me0)2POCH2CO2Me
NC 0 vo".0Ac 3. K2CO3
OH LiCI, i-P
4. HC!; Na104 0
0 2NEt
ER818638
ER-818636 crystalline
major anomer
H
H
./.--0
H2/Pd/C T120, NEt3
NC0"----1--\--\ CO2Me NCCY---)--.\---0O2Me
'-.'
ER-818640 .0H ER-818641 0H
H
H
Na! LiBH4 or NaBH4
NC 0 CO2Me _____________ NC 0 CO2Me ____
.0Tf Acetone 1 THF
ER-818649 ER-818642
H
.-"----0 OH
Zn/Et0H ______________ Of---- /
0---'".---"',-;3_2
Ne0'----)---"\ __ \OH
I 7
ER-818643 ER-818644
TBDPSCI Of---- /0TBDPS
0----./"j.
ER-813012
19

CA 02822994 2013-08-01
Scheme V-c
--_
o--- OTBDPS
O----' OTBDPS
0--"\----; / LDA, Mel
-
ER-813012 ER-817704
1. NH(OMe)Me HCI i i OTBS OTBDPS
AlMe3 / MeMgCI
Me0
2 TBSCI 0
ER-806728
= OTBS OTBDPS f. OTBS OTBDPS
/ KHMDS /
;1-13_2
0 Tf2NPh OTI
ER-806727 ER-806729
= OH OH
HCI = = --i3--' ____ /
;70)-.2
_
IPA OTf
ER-806730
[0056] Using ER-817664 as a crystalline intermediate, Scheme VI-a shows a
general
method for using this compound in the preparation of intermediates useful for
preparing
compounds of formula F-2.

CA 02822994 2013-08-01
Scheme VI-a
o /",, 0
BrCOC(CH3)20Ac DBU
NCCIµµµ
"'OH '"OAc
ACN, 0 C
0H Br
DMP, H+
1 03, Me0H
_________________________________ ¨ NC \--
NCO'sµ ."0Ac 2 NaBH4 OH
OH
NIS, PPh3
NC-0"µ
OH
0
Zn, Et0H
80 C
ER-812835
[0057] The triol intermediate depicted in Scheme VI-a above is used in an
alternate
method for preparing intermediates useful for preparing compounds of formula F-
2, as
shown in Scheme VI-b below.
21

CA 02822994 2013-08-01
Scheme VI-b
CN ji pH Na104, THE CN Ph3PCHCO2Me
___________ CHO
0OH pH 7 buffer THF, 50 C
(:)H
OH
CN H2
CN
Pd/C
v-j¨

0O2Me Et0Ac 0 CO2Me
OH OH
N-iodosuccinimide CN 0 NaBH4
PPh3, THE 0 CO2Me Et0H, H20
0
Zn
0 Et0H "=-=5 OH
OH
1
C.14 C.26 lactone
[0058] In Scheme VI-b, shown above, the triol intermediate is treated with
periodate to
form the aldehyde. This compound is homologated with methyl Wittig reagent,
and the
resulting olefin reduced, to form the ester compound. The remaining free
hydroxyl group is
treated with N-iodosuccinimide to form the iodo intermediate and the ester
reduced with
sodium borohydride to form the hydroxyl compound depicted above. One of
ordinary skill
in the art will recognize that the resulting iodo compound corresponds to
compound 21
depicted in Scheme II, supra, wherein compound 21 has a protecting group at
the hydroxyl
position. The final treatment with zinc affords the lactone depicted above.
One of ordinary
skill in the art will recognize that the resulting lactone compound
corresponds to compound
22 depicted in Scheme II, supra, wherein compound 22 has a protecting group at
the
hydroxyl position.
[0059] Yet another alternate method for preparing intermediates useful for
preparing
compounds of formula F-2 from D-quinic acid provides an alternative route from

intermediate 2, of Scheme II as shown in Scheme VII below.
22

CA 02822994 2013-08-01
Scheme VII
HO'
,
+ CIBr Pyridine, 0 C
B1 A0.

' 2. DBU, 80 C
'sl 0 0 __ PPh3MeCN
(a) (b)
ER-811510 ER-812771
0 n
0 = =-= ,H
õH
<zy H
_______________________________ 0 0
0 (c) Pd/C, H2 "'0
Ph3P "0
ER-812772 ER-812829
[0060] In Scheme VII above, intermediate 2 (from Scheme II) is used to
prepare ER-
812829 in a stereoselective manner. It will be appreciated that other
protecting groups are
useful for protecting the diol of ER-812829. Such groups are known to one of
ordinary skill
in the art and include cyclohexylidene and benzylidene diol protecting groups.
First, at step
(a), the hydroxyl group of ER-811510 is treated with 2-bromo acetylchloride to
form ER-
812771. The bromo intermediate is treated with triphenylphosphine to form a
Wittig reagent
in situ in a manner substantially similar to that described by Murphy, et al,
Tetrahedron
Letters, 40, (1999) 3455-3456. This Wittig reagent then forms the lactone ER-
812772. At
step (d), stereoselective hydrogenation of the double bond affords ER-812829.
[0061] The present invention also provides a method for preparing
intermediates useful
for preparing compounds of formula F-2 from D-quinic acid, from intermediate
ER-812829
depicted in Scheme VII above, as shown in Scheme VII-a below.
23

CA 02822994 2013-08-01
Scheme VII-a
HO-7'0 TsCI Ts0----/''' Cs)
HO"'
.0
---0"------ -'o---- --"0------
ER-820100 ER-812897 ER-812880
7 0 7 OH
_
=_
1 r6 TBSO H .,,,-.,,,,,õ-= TBSO 7
0
Nat E
- HO"' . =
"0 HO .
Cr L* Co, TMSCI
b-"--- Mn '
ER-812881 ER-820098
..õ
Dess-Martin
TBSO.,. Et3SiH TBS0.0õ.0
periodane: HO
- ."0 BF3-0Et2 7
b---- -b------
ER-812884 ER-812909
/OFI
V
ER-806730
3. Fragment F-3
[0062] According to yet another embodiment, the present invention provides
a compound
F-3:
oPG4
0 ---
LG2
R3
o oPG4
oPG4
F-3
wherein:
each PG4 is an independently selected suitable hydroxyl protecting group;
R3 is CHO or C(0)0R4;
R4 is a suitable carboxyl protecting group; and
LG2 is a suitable leaving group.
24

CA 02822994 2013-08-01
[0063] Suitable carboxylate protecting groups are well known in the art and
are described
= in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P.
G. M. Wuts, 3rd
edition, John Wiley & Sons, 1999. In certain embodiments, the R4 group of F-3
is an
optionally substituted Ci_6 aliphatic group or an optionally substituted aryl
group. Examples
of suitable R4 groups include methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, benzyl, and
phenyl wherein each group is optionally substituted.
[0064] As described above, suitable leaving groups are well known in the
art, e.g., see
"Advanced Organic Chemistry," Jerry March, 4th Ed., pp. 351-357, John Wiley
and Sons,
N.Y. (1992). Such leaving groups include, but are not limited to, halogen,
alkoxy,
sulphonyloxy, optionally substituted allcylsulphonyloxy, optionally
substituted
alkenylsulfonyloxy, optionally substituted arylsulfonyloxy, silyl, and
diazonium moieties.
Examples of suitable leaving groups include chloro, iodo, bromo, fluor ,
methanesulfonyloxy (mesyloxy), tosyloxy, triflate, nitro-phenylsulfonyloxy
(nosyloxy), and
bromo-phenylsulfonyloxy (brosyloxy). In certain embodiments, the LG2 moiety of
F-3 is
iodo.
[0065] According to an alternate embodiment, the suitable leaving group may be

generated in situ within the reaction medium. For example, LG2 in a compound
of formula
F-3 may be generated in situ from a precursor of that compound of formula F-3
wherein said
precursor contains a group readily replaced by LG2 in situ. In a specific
illustration of such
a replacement, said precursor of a compound of formula F-3 contains a group
(for example,
a trimethylsilyl group) which is replaced in situ by LG2, such as an iodo
group. The source
of the iodo group may be, e.g., N-iodosuccinimide. Such an in situ generation
of a suitable
leaving group is well known in the art, e.g., see Id.
[0066] As described above, suitable hydroxyl protecting groups are well
known in the art
and include those described in detail in Protecting Groups in Organic
Synthesis, T. W.
Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999.
In certain embodiments, each PG4, taken with the oxygen atom to which it is
bound, is
independently selected from esters, ethers, silyl ethers, alkyl ethers,
arylalkyl ethers, and
alkoxyalkyl ethers. Examples of such esters include formates, acetates,
carbonates, and
sulfonates. Specific examples include formate, benzoyl formate, chloroacetate,

trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-
chlorophenoxyacetate, 3-
phenylpropionate, 4-oxopentanoate, 4,4-(ethylenedithio)pentanoate, pivaloate
(trimethylacetyl), crotonate, 4-methoxy-crotonate, benzoate, p-benzylbenzoate,

CA 02822994 2013-08-01
2,4,6-trimethylbenzoate, carbonates such as methyl, 9-fluorenylmethyl, ethyl,
2,2,2-
trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl,
allyl, and p-nitrobenzyl.
Examples of such silyl ethers include trimethylsilyl, triethylsilyl, t-
butyldimethylsilyl, t-
butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl ethers. Alkyl
ethers include
methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, ally!,
and
allyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers include acetals
such as
methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl, benzyloxymethyl,
beta-
(trimethylsilypethoxymethyl, and tetrahydropyranyl ethers. Examples of
arylallcyl ethers
include benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, o-nitrobenzyl, p-
nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2- and 4-
picolyl.
[0067] In certain embodiments, one, two, or three of the PG4 moieties of F-
3, taken with
the oxygen atom(s) to which they are bound, are silyl ethers or arylallcyl
ethers. In yet other
embodiments, one, two, or three of the PG4 moieties of F-3 are t-
butyldimethylsilyl or
benzyl. In still other embodiments, all three of the PG4 moieties of F-3 are t-

butyldimethylsilyl.
[0068] According to another embodiment, a compound of formula F-3 is provided
wherein said compound has the stereochemistry as depicted in formula F-3':
oPG4
H
LG2
4
OPG
= oPG4
F-3'
wherein each variable is as defined above and described in classes and
subclasses above and
herein.
[0069] In certain embodiments, a compound F-3a is provided:
OTBS
oL7
Me02C0--
17-1
OTBS
F-3a
wherein "TBS" refers to t-butyldimethylsilyl.
[00701 Details of the synthesis of F-3a are set forth in the Examples
infra.
26

CA 02822994 2013-08-01
4. Assembly of F-1, F-2, and F-3 to Prepare Compound I
[00711 Coupling of the fragments F-1 and F-2 is accomplished, in general,
as set forth in
Scheme VIII below.
Scheme VIII
SO2Ph
PG20 R. _________________________________________ ("SO2Ph
µ.µ40)=R2
, F-1 CrC12, NiCl2 c
0PG- HO'rj
OPG1 THF, Et3N
LG1
=
LG1 X
O
NHMs PG3
ER-807363
0 F-4
F-2
R,
PG20
1-SO2Ph ("SO2Ph
PG20
PG1OL),,,
0
0
=,õ
I 0I 0
F-5a F-5
[00721 Scheme VIII above shows a general method for preparing intermediate
F-5a from
fragments F-1 and F-2. First, fragments F-1 and F-2 are coupled using methods
substantially similar to that described by Kishi, et al., Org Lett 4:25 p
4431(2002) to afford
intermediate F-4. This coupling is performed in the presence of the chiral
oxazole (ER-
807363) or, alternatively, in the absence of ER-807363. However, the coupling
reaction of
F-1 and F-2 proceeds with higher selectivity when performed in the presence of
ER-807363.
Intramolecular Williamson ether formation of F-4, by treating F-4 with
potassium
hexamethyldisilazide, then furnishes tetrahydropyran F-5 as a mixture of
stereoisomers. The
stereoisomers are then separated to afford F-5a. The details of these steps
are set forth in the
Examples infra.
=
27

CA 02822994 2013-08-01
[0073] According to another embodiment, the present invention provides a
compound
F-4:
R1
pG20 CO2Ph
PG10
0
HO
LG1
OPG
F-4
wherein PG', PG2, PG3, LGI, and RI are as defined in general and in subclasses
above and
herein.
[0074] In certain embodiments, the present invention provides a compound of
formula F-
4 wherein said compound has the stereochemistry depicted in formula F-4':
R,
PG20 SO2Ph
ci
PG10,õ=
0
HO'r
LG1
0
ye.OPG3
F-4'
wherein PG', PG2, PG3, LGI, and RI are as defined in general and in subclasses
above and
herein.
[0075] The present invention also provides a compound F-4a:
28

CA 02822994 2013-08-01
Me0,SO2Ph
TBSO
HO
TBSO,,),õsc
0
OMs
F-4a
wherein "Ms0" refers to mesylate, "TBS" refers to t-butyldimethylsilyl, and
"OPv" refers to
pivaloate.
[0076] Details of the synthesis of F-4a are set forth in the Examples
infra.
[0077] According to yet another embodiment, the present invention provides
a compound
F-5:
R1
pG20 õCSO2Ph
PG10
0
0
)0PG3
F-5
wherein each PG'. PG2, PG3, and R1 is as defined in general and in subclasses
above and
herein.
[0078] In certain embodiments, the present invention provides a compound of
formula F-
having the stereochemistry as depicted in formula F-5' or F-5a:
G20
R,
,
( R
____________________ --- PG20 -SO2Ph 1---802Ph
P ci
PG1O
PG10õ,õsco)=.õ
0
0
IIooPG3
F-5' F-5a
29

CA 02822994 2013-08-01
wherein each PG', PG2, PG3, and RI is as defined in general and in subclasses
above and
herein.
[0079] The PG3 group of intermediate F-5a is removed and the resulting
hydroxyl
compound F-6 is then coupled with a compound F-3', wherein R3 is CHO, to form
F-7 as
depicted in Scheme IX below.
Scheme IX
1
PG20 R _____________ ¨SO2Ph
- so2ph
PG2o
s 0
OH 0
F-6 I o 0 H
y"OH C/-
1G40E-10PG4
OPG4 \\\\
oOH Li
G2
OPG4
H 0 y LG2
0 F-7
OPG4
171
OPG4
F-3' so2P6
R,
PG20
0 0
mõ, 0
1
R,
PG20
OPG4
H OPG4
= CO 0,0
I 0
OPG4 LG2
PG .0
H OPG4
F-8
=
Lo2
F-9
[00801 Scheme IX above shows a general method for preparing an
intermediate F-9 from
F-3' and F-6. First, the sulfone intermediate F-6 is treated with n-butyl
lithium then with
the aldehyde F-3'. The resulting diol intermediate F-7 is then oxidized with
Dess-Martin
reagent to form the ketone-aldehyde intermediate F-8 which is then treated
with SmI2 to
afford intermediate F-9. The details of these steps are set forth in the
Examples infra.

CA 02822994 2013-08-01
Scheme X
R! RI,
PG2o pG2o
PG10sõ=Co 0 0 0 0
0 0 0 0
1-1,0PG4
PG-0 PG40
.fõ H OPG4 OPG4
OH
F-9a F-10
OH _____
PG20
=Co =.õ: 0 0 PG10), =Co 0 0
ss,
sõ
o 0
OH H'
0
OPG4 HO 0 PG40
sss, 0 ss=k../"=,, OPG4
I 0 I,,,, 0
õ,.
0 0
F-12 F-11
[0081] Scheme X above sets forth a general method for preparing the
Halichondrin B
analogs of the present invention from F-9a (LG2 is iodo). First, an
intramolecular coupling
is achieved, by conditions substantially similar to those described at Scheme
V above, to
form hydroxyl compound F-10. In an alternate method, the intramolecular
coupling is
performed in the presence of the chiral oxazole ligand, described herein. The
addition of the
chiral oxazole ligand imparts a higher yield and greater efficiency for the
reaction. The
details of this reaction are set forth in the Examples below. Compound F-10 is
then oxidized
to form F-11. The hydroxyl protecting groups of F-11 are removed by
appropriate means to
afford F-12. One of ordinary skill in the art would recognize that the methods
appropriate to
achieve removal of the protecting groups of compound F-11 depend upon the
actual
protecting groups used and include those described by Greene. For example,
when each of
the hydroxyl protecting groups of F-11 is a TBS group, such removal may be
achieved by
treatment with optionally buffered tetrabutylammonium fluoride. The details of
these steps
are set forth in the Examples infra.
[00821 Intermediate F-12 is useful for preparing various analogs of
Halichondrin B as
described in, e.g. United States Patent 6,365,759 and United States Patent
6,469,182.
31

CA 02822994 2013-08-01
EXAMPLES
[00831 Using the preparation of Halichondrin B analog B-1939 to exemplify,
the
following Examples describe the synthesis of Halichondrin B analogs using the
methods and
compounds of the present invention.
MeQ.,
HO
H 2N s,
0 ""'¨ =
0
s
=I Q O.
C),,
1,õ 0
B-1939
[00841 One of ordinary skill in the art would recognize that many analogs
of
Halichondrin B are prepared by the methods and from the compounds of the
present
invention including, but not limited to, those analogs of Halichondrin B
described in United
States Patents 6,214,865 and 6,365,759.
Accordingly, it will be appreciated that the synthetic methods described
below, by way of
example, do not limit the scope of the invention which is defined by the
appended claims.
Example 1
Preparation of F-la:
HO H
0 H 01\
= 0 cat. H2SO4 0
0 OH _____________
0
0 = - Acetone, ACN, 500HOH C
HO H
ER-806044 ER-806045
[00851 In an appropriately sized vessel, D-glucurono-6,3-lactone (1 wt., 1
eq.) was
combined with ACN (3 vol.) and acetone (9 vol.). Catalytic conc. sulfuric acid
was added
and the system held at reflux for 3 hours. The system was checked for
dissolution of
D-glucurono-6,3-lactone. The reaction was cooled to 25 C and stirred for 15
hours. Solid
sodium bicarbonate (0.5 wts) was added and the reaction stirred for 3
additional hours.
Solids were removed by filtration and the organics were partially concentrated
and
azeotroped with additional ACN (2 wts). ER-806045 was taken into the next
reaction
without isolation.
32

CA 02822994 2013-08-01
El oi\
0 0
SO2C12, Pyne,
0 ' 0
F10 H ACN, 80% CI 0
ER-806045 ER-806410
[0086] Crude ER-806045 (1 wt, 1 eq.) was dissolved in ACN (6.5 vol.) at -20
C.
Pyridine (1.5 vol., 4.0 eq.) was added and S02C12 (0.38 vol., 1.02 eq.) was
added slowly,
while keeping the internal temperature below 5 'C. The reaction was quenched
by inverse
addition into cool water (28 vol.) with an ACN rinse (0.5 vol.), keeping the
internal
temperature below 10 'C. The white solid, ER-806410 (0.87 wt., 79% of
theoretical) was
isolated by filtration with a heptane rinse (2 vol.) and drying.
0 H 0 j
0
H2, Pd (carbon)
CI H THF, 74%
ER-806410 ER-806047
[0087] An appropriately sized vessel was charged with ER-806410 (1 wt, 1
eq.) and THY
(10 vol.) and then cooled to 10 C. Wet palladium on carbon (5%, 0.5 wts) was
added and
the heterogeneous solution stirred for ten minutes. The reaction was buffered
with pyridine
(0.44 wts, 1.3 eq.) and placed under a hydrogen atmosphere for 3 hours. The
reaction was
filtered and the solids rinsed with water (2 vol.) and Et0Ac (10 vol.). The
resulting solution
was acidified with 1N HC1 (2.1 vol.), mixed well and the resulting layers were
separated.
The organic layer was sequentially washed with aqueous sodium bicarbonate (5
vol.) and
water (5 vol.). The organics were concentrated under reduced pressure and the
resulting
product recrystallized from IPA (3.4 vol.) and further cropped by the addition
of heptane
(3.4 vol.) at 15 'C. ER-806047 was isolated as a white solid (67% yield).
0 0,,
DIBAL, Toluene __ ,( (
)-0
-40 C 80% 0
ER-806047 ER-806048
[0088] An appropriately sized vessel was charged with ER-806047 (1 wt, 1
eq.) and
toluene (8 vol.) and then cooled to -40 C. A 17 wt% solution of DIBAL in
toluene (4.6 wts,
1.1 eq.) was added, keeping the internal temperature below -35 C. After
assaying the
reaction, excess reagent was quenched by the addition of acetone (.15 wts, 0.5
eq.), keeping
the temperature below 10 C. The reaction was diluted with Et0Ac (7 vol.) and
15%
aqueous citric acid (8 wts) below 10 C, and stirred at 20 C until a clear
solution was
obtained. The
33

CA 02822994 2013-08-01
layers were separated and the aqueous layer back extracted twice with Et0Ac (2
x 10 vol.).
The combined organics were washed sequentially with aqueous sodium bicarbonate
(5 vol.)
and brine (5 vol.) and then dried with magnesium sulfate (0.2 wts). After
filtration, the
organic layers were partially concentrated at reduced pressure, and azeotroped
with toluene
(4 vol.). The products were stored as a THF solution for use in the next
reaction.
ic.1\ TMSCH2MgCl. HR
o)--O THF, 35 C, 1 5h TMSC OH
-S-- 0
0
ER-806048 ER-807114
[0089] An appropriately sized vessel was charged with a 20 wt% ether
solution of
TMSCH2MgC1 (2.04 wts, 3.0 eq.) and chilled below 5 'C. A THE (7 vol.) solution
of ER-
806048 (1 wt, 1 eq.) was added to the reaction vessel keeping the internal
temperature below
15 C. The reaction was warmed to 35 C for 1.5 hours. The reaction was
cooled, diluted
with toluene (7 vol.) and quenched with AcOH (3 vol.) below 20 C. The
reaction was
further diluted with 10% aqueous ammonium chloride (6 wts), mixed well, and
the layers
were separated. The organic layer was washed sequentially with aqueous sodium
bicarbonate (5 vol.) and brine (5 vol.). After drying over magnesium sulfate
(0.2 wts) and
filtration, the solution was concentrated under reduced pressure and ER-807114
isolated as a
concentrated solution in toluene (90% yield).
HR HO
e01\
OH ICHMDS
0
õ
" 0 THF, rt 0
ER-807114 ER-806049
[0090] An appropriately sized vessel was charged sequentially with ER-
807114 (1 wt, 1
eq.) and THF (20 vol.), and the solution was cooled below 5 C. A 15 wt%
solution of
KIIMDS in toluene (9.16 wts, 2.0 eq.) was added. The reaction was quenched
with 10%
aqueous ammonium chloride (5 vol.). The layers were separated and the organic
layer
washed sequentially with ammonium chloride (5 vol.), 2N HCI (8.5 vol.),
aqueous sodium
bicarbonate (5 vol.), and water (5 vol.). The organics were transferred to
concentration
vessels using Et0Ac, and concentrated to a viscous oil (90% yield). The
material was
recrystallized from toluene (4 vol.) and heptane (4 vol.) at 35 C with
additional cropping at
lower temperatures and heptane (2 x 4 vol.) at 15 and 10 C (94% yield).
34

CA 02822994 2013-08-01
HO, 0 Bn0
BnBr, KOtBu,
TILE', 70% two steps
ER-806049 ER-806050
[0091] An appropriately sized vessel was charged with KOtBu (0.67 wts, 1.2
eq.) and
TI-IF (7.7 vol.), and cooled to an internal temperature of -20 C. A solution
of ER-806049 (1
wt, 1 eq.) in THF (2.3 vol.) was added keeping the internal temperature below -
7 C. Neat
BnBr was added, maintaining -7 C as the maximum temperature. The reaction was
stirred at
-20 C for 2 hours and 10 hours at 10 C. The reaction was quenched with 10%
aqueous
NH4C1 (4 wts), diluted with toluene (4 vol.), and mixed well. The layers were
separated and
the organic layer washed with 10% brine (4 wts) and dried over MgSO4 (0.15
wts). ER-
806050 was isolated as a tBuOH solution (2.5 vol.) after concentration at
reduced pressure
(95% yield).
0
BnO, 01\ (DHQ)2AQN, BnO,
K20s04 2H20, HO
0

HO 0
s'.."--
K3Fe(CN)6, K2CO3 ) ER-806051
ER-806050 tBuOH:H20 3:1 ER-806052
[0092] An appropriately sized vessel was charged sequentially with
K3Fe(CN)6 (3.5 wt.,
3.4 eq.), K2CO3 (1.5 wt., 3.4 eq.), (DHQ)2AQN (0.0134 wt., 0.005 eq.), water
(18 vol.), t-
BuOH (13 vol.), and ER-806050 in tBuOH (1 wt, 1 eq. in 5 vol.). The
heterogeneous
mixture was cooled to an internal temperature of 0 C, and K20s04-2H20 (0.0029
wt., 0.22
mole%) was added. After 36 hours at 0 C, the reaction was quenched with
Na2S203 (3.5 eq.,
1.7 wt.) and the flask allowed to warm to ambient temperature overnight. After
15 hours, the
mixture was transferred to a workup vessel and diluted with toluene (15 vol.)
and water (4
vol.). The biphasic mixture was vigorously stirred and separated. The organic
layer was
washed with brine (10 vol.), and concentrated and solvent exchanged to afford
a crude
mixture of diols ER-806051 and ER-806052 as a 10% toluene solution (92%
yield).
BnO, 0
BzCI, N-methylmorpholine,
HO Bz0
DP, PhCH; BzO
C3:1 ER-806051 C 3:1 ER-806053
ER-806052 ER-806054

CA 02822994 2013-08-01
[0093] The toluene solution (10.1 wt%, 9.9 wts) of ER-806051/52 (1 wt, 1
eq.) was
further diluted with additional toluene (3 wts). N-Methylmorpholine (0.94 wts,
3.0 eq.) and
DMAP (0.075 wts, 0.2 eq.) were added to the toluene solution and the resulting
mixture was
cooled below 15 C. Benzoyl chloride was added keeping the intemal temperature
below 25
C. The reaction was then stirred for 12 hours at 75 C. The reaction was
cooled to 15 C and
the temperature kept below 25 C during the 1N 1-ICI (5 vol.) quench. Layers
were mixed
well and separated. The organic layer was sequentially washed with brine (3
wts), aqueous
sodium bicarbonate (3 wts), and brine (3 wts). The organic layer was dried
(MgSO4, 0.25
wts), treated with activated carbon (0.1 wts), and filtered (Celite , 0.3 wts)
with toluene (1
wt). The products were partially concentrated under reduced pressure,
azeotroped with
toluene (3 wts). Bisbenzoate ER-806053/54 was isolated in 95% yield as a
toluene solution
(5 vol.).
OH
0
TiC13(0iPr), Ally1TMS,
3:IrL toluene, 30 'V
OBz
OBz ER-806053 OBz ER-806055
ER-806054
[0094] Under an inert atmosphere, a 20 wt% solution of TiC14 (6.42 wts, 3.6
eq.) in
toluene was cooled to 15 'C. Keeping the internal temperature below 30 C,
Ti(OiPr)4 (0.64
wts, 1.2 eq.) was added, and the resulting solution stirred for 15 minutes.
A11y1TMS (1.03
wts, 4.8 eq.) was premixed with ER-806053/54 (1 wt, 1 eq.), available as a 22
wt% solution
in toluene from the previous step (4.55 wts, 1 eq.), and added to the freshly
generated
Ti(OiPr)C13. The internal temperature during the addition was kept below 30 C.
The
reaction was stirred between 20-30 C for 2 hours. The reaction was cooled to -
5 C and
quenched with IN HC1 (6 vol.), keeping the internal temperature below 30 C.
After mixing
well, the layers were separated and the organic layer sequentially washed with
IN HC1 (3
vol.) and brine (2 x 3 vol.). The organic layer was stirred with MgSO4 (0.3
wts) and
activated carbon (0.15 wts) and filtered through a Celite plug (0.2 wts),
rinsing with
toluene (1 vol.). The product, as a 3:1 mixture at C-34 was isolated after
concentration in
83% yield. Recrystallization from IPA/n-heptane afforded ER-806055 with >99.5%
d.e. at
C-34 (71% yield).
36

CA 02822994 2013-08-01
OH 0
Bn0õ,
µ...
s' ¨ DMSO, TCAA, Et3N
o 0
Toluene, <-10 C
OBz OBz
ER-806055 ER-806058
[0095] At room temperature, an appropriately sized vessel was charged with
alcohol
ER-806055 (1 wt., 1.0 eq.), toluene (7 vol.), DMSO (0.31 wt., 2.0 eq.) and
Et3N (0.78 wt.,
4.0 eq.). The resulting solution was cooled to -19 C. TCAA (0.84 wt., 1.4
eq.) was added
drop-wise, keeping the internal temperature below -10 C. The reaction was
stirred for an
additional 10 minutes. The reaction was diluted with IPA (0.5 vol.) and
quenched with IN
HCI (5 vol.), keeping the internal temperature below 10 C. The layers were
separated and
the organic layer sequentially washed with aqueous NaHCO3 (5 wt.) and water (3
vol.). The
organic layer was partially concentrated at reduced pressure (100% crude
yield) and further
azeotroped with additional toluene (4 vol.). The resulting ketone (ER-806058)
was dissolved
in a final 4 vol. of toluene, checked for water content and used as is in the
next reaction.
Pho2s
0 ,j 10-11:1
ER-107446, LH1VIDS
ToluenefTHF,
C to RT c)'''OBz
OBz ER-806058 OBz ER-806059
[0096] A solution of ER-107446 (1 wt, 1.5 eq.) in THF (2.7 vol.) was cooled
to 10 C
and treated with 25.5 wt% LHMDS in THF (5.2 wt., 1.4 eq.), keeping the
internal
temperature below 15 'C. In a second vessel, the toluene solution of crude ER-
806058 (21.9
wt%, 5.4 vol.) was cooled to 10 'C. The contents of vessel one were
transferred into the
substrate containing solution, keeping the internal temperature below 20 'C.
The reaction
was stirred for 30 minutes then quenched by adding 1M HCI (6.5 vol.), keeping
the internal
temperature below 20 'C. The layers were separated and the organic layer
washed four times
with 1:1 Me0H/water (4 x 5 vol.) and then with aqueous bicarbonate (5 vol.),
and brine
solutions (2 x 5 vol.). The product was dried over MgSO4 (0.52 wt.), filtered
(rinsing with
0.7 vol. of toluene), and concentrated to a heavy oil at reduced pressure.
37

CA 02822994 2013-08-01
PhO2S PhO2S
TMSI, CH3CN/toluene HO
\¨ 60 C
,=== 0
"OBz "OBz
OBz OBz
ER-806059 ER-806060
100971 ER-806059 was dissolved in 1:1 toluene/CH3CN (5 vol.) at room
temperature.
Filtered TMS1 (1.23 wt., 4 eq.) was added, keeping the initial temperature
below 40 C. The
reaction was heated to 60 C for 2 hours. The reaction was cooled to -15 and
quenched with
25% aqueous ammonium hydroxide below 30 C. The reaction contents were stirred
overnight and the layers separated. The organic layer was charged with
additional toluene (5
vol.) and water (2 vol.). The layers were mixed well and separated. The
organic layer was
then washed sequentially with 10% aqueous sodium sulfite (5 vol.), IN HC1 (5
vol.), 5%
aqueous sodium bicarbonate (5 vol.), and brine (5 vol.). The organic layer was
dried over
MgSO4 (0.2 wt.), filtered, partially concentrated and used in the next
reaction as a 50%
solution in toluene.
PhO2S
so2Ph
HO., 4' HO.,.
Bu4NCI, NaBH(OAc)3
DMETToluene, 85 C
r)'''OBz
OBz OBz
ER-806060 ER-806061
[0098] In an appropriately sized vessel, NaBH(OAc)3 (1.19 wt, 3.15 eq.),
Bu4NC1
(1.04.wt, 2.1 eq.), DME (8.2 vol.), and toluene (4 vol.) at 65 C, were
combined and stirred at
room temperature. The mixture was heated to 75 C for one hour. ER-806060 (1
wt., 1 eq.)
as a 50% wt solution in toluene was added at 75 C and rinsed in with
additional toluene
(0.3 vol.). The reaction temperature was raised to 85 C and the reaction
stirred for 2-4
hours. The reaction was cooled to < 10 C and quenched with water (3.2 vol.)
keeping the
internal temperature below 20 C. The layers were mixed well and separated.
The organic
layer was sequentially washed with aqueous sodium bicarbonate (2 x 5 vol.) and
water (2 x 5
vol.). The organic layer was concentrated and solvent exchanged to afford a 40
wt% solution
of ER-806061 in Me0H.
38

CA 02822994 2013-08-01
SO2Ph
HOõ,
1. K2CO3, Me0H, 50 C HO
2. recrystallization, n-BuOH
57% five step yield
"oH
OBz OH
ER-806061 ER-806064
[0099] A 40 wt% solution of ER-806061 (1 wt., 1.0 eq.) in Me0H was
dissolved in
additional methanol (1.6 vol.). Potassium carbonate (0.24 wts, 1.0 eq.) was
added and the
reaction temperature was raised to 50 C for one hour. The reaction was cooled
to 15 C,
and quenched with 1N FICI (3.5 vol., 2 eq.) with the internal temperature
below 30 C. The
reaction was diluted with water (3.9 vol.) and toluene (3 vol.). The layers
were separated and
the aqueous layer back extracted with toluene (1.5 vol.). The aqueous phase
was charged
with sodium bicarbonate (0.3 wts) and sodium chloride (0.6 wts), and back
extracted with
nBuOH (3 vol.). The three organic phases were combined and concentrated to
dryness to
afford crude trio! ER-806064 and inorganic salts. The product was dissolved in
7:1
toluene/nBuOH at 80 C, hot filtered, and recrystallized by cooling and
stirring overnight.
ER-806064 (F-1b) was isolated in a 57%, five step overall yield after
filtration and a toluene
rinse. FAB(+)-MS m/z 357 (M + I-1). Melting point 96 .2 C.
so,Ph SO2Ph
HOõ,
(CH3)2C(OMe)2, H2SO4,
Acetone, 25 C
OH
ER-806064 I ER-806126
[00100] Purified triol ER-806064 (1 wt., 1 eq.) was dispersed in acetone (2
vol.), diluted
with 2,2-dimethoxypropane (1 vol.), and treated with conc. sulfuric acid
(0.0086 wts, 0.03
eq.) at 25 C. The reaction was stirred until homogenous. The reaction was
diluted with
toluene (5 vol.) and quenched by the addition to 5% K2CO3 (2 vol.). The layers
were mixed
well and separated. The organic layer was washed with 10% brine, dried with
Na2SO4 (0.5
wt.). The solution was filtered (toluene rinse) and concentrated at reduced
pressure to afford
ER-806126 as a yellow oil. The material was used as is in the next stage.
39

CA 02822994 2013-08-01
SO2Ph SO2Ph
HOõ, Me0,
NaOtBu, Mel
THF, 15-25 C
µ- 0 \
(1..0 (0
ER-806126
ER-806068
[001011 Solid NaOtBu (0.34 wt., 1.4 eq.) was dissolved in THF (2.7 vol.)
and DMF (0.3
vol.), and then cooled below 10 C. A solution of ER-806126 (1 wt., 1 eq.) in
TI-IF (2.5 vol.)
was added to the NaOtBu solution with a THF rinse (0.5 vol.), keeping the
internal
temperature below 15 C. After a 30 minute stir, methyl iodide (0.204 vol., 1.3
eq.) was
added keeping the temperature below 15 C (exothermic). The reaction was warmed
to 25 C
and the reaction quenched with water (5 vol.) and diluted with toluene (7
vol.). The layers
were mixed well and separated. The organic layer was washed twice with brine
(2x5 vol.),
dried over Na2SO4 (0.5 wt.), filtered, and concentrated under reduced
pressure.
so2Ph SO2Ph
2M HCI, Me0H,
"S ____________________________________ p
¨ ss' 0
25 'C
(
OH
ER-806068 ER-806063
[001021 ER-806068 (1 wt., 1 eq.) was dissolved in 1 vol. of Me0H. Water
(1.5 vol.) and
2 N HCI (1.25 vol., 1 eq.) were added and the reaction stirred at 25 C. The
reaction was
quenched by inverse addition to 2M NaOH (1.34 vol.) at 10 C. The reaction was
diluted
with isopropyl acetate (5 vol.), the layers were mixed well and separated. The
aqueous layer
was back extracted with 5 vol. of isopropyl acetate and the combined organic
layers were
dried over MgSO4 (0.5 wt.), filtered, and concentrated at reduced pressure to
afford crude
diol ER-806063.
so2Ph SO2Ph
Me0õµ Me0õ,
TBSCI, Imidazole
DMF
so 0
ER806063 d'''OTBSER806065
OH C18H2606S OTBS C301-15406SS12
Mol. Wt.: 370.46 Mot. Wt.: 598.98
[001031 To a 25 C solution of crude ER-806063 (1 wt., 1.0 eq.) in DMF (4
vol.), was
charged imidazole (0.62 wt., 3.4 eq.), followed by TBSC1 (1.02 wt., 2.53 eq.)
with the
' 40

CA 02822994 2013-08-01
internal temperature below 30 C. The reaction was stirred at 25 C. The
reaction was diluted
with MTBE (10 vol.) and washed with I-120 (4 vol.). The organic layer was
washed
sequentially with 1M 1-IC1 (3 vol.), water (3 vol,), aqueous sodium
bicarbonate (3 vol.), and
brine (3 vol.), The organic layer was dried over MgSO4 (0.5 wt.), filtered
with one volume
MTBE rinse, and concentrated at reduced pressure and solvent exchanged for
heptane (4
vol.).
Me0,( __________ SO2Ph IVIe0, (---S02Ph
õ
1.03, Heptane -50 C
2. Warm to 5 C
3_ Lindlar cat, H2 (g)
r-OTBS r-OTBS
OTBS ER-806065 RT, 2.5 hours
OTBS ER-806067
[001041 ER-806065 (1 wt., 1 eq.) was dissolved in heptane, isooctane, or
IPA (10 vol.)
The solution was cooled below -60 C ( 10 C). Ozone was bubbled through the
solution at
low temperature until the solution retained a blue color. Nitrogen was purged
through the
solution for 15-30 minutes, and the reaction warmed to 5 C while the nitrogen
flush was
continued. 7-15 wt. % Lindlar Catalyst (5% Pd on CaCO3 poisoned with Pb, 0.1
wt.) was
added. The reactor head was purged several times with nitrogen, evacuated, and
placed
under 1 atmosphere H2 (g). The reaction was then warmed to room temperature
(20-25 C).
The reaction was stirred for 2.5 hours. The resulting heterogeneous solution
was filtered
through Celite (1.0 wt.) with an MTBE (2 vol.) rinse. The solution was
concentrated to
dryness, isolating 1.0 wt. of crude ER-806067. The crude isolate was
recrystallized from
heptane or isooctane to afford ER-806067 (F-1a) as a white crystalline solid
in a 68% five
step yield. FAB(+)-MS m/z 601 (M + H). Melting point 64.5 C.
Example 2
Preparation of F-2a:
Br
OH
Ambedyst ç00H Br
O
Br H
H20, 5 C Sn, HBr, H20
ER-806909
35 C
[00105] A reactor was charged with pre-rinsed Amberlyst 15 (0.05 wt.) and
water (4.63
vol.) and cooled to an internal temperature of 0-5 C. The reactor was charged
with 2,3-
dihydrofuran (1 wt., 1 eq.) and stirred for 1.5 hours maintaining internal
temperature around
41

CA 02822994 2013-08-01
C. A second reactor was charged with water (4.63 vol.) and heated to an
internal
temperature of 35 C. The same reactor was charged with tin powder (2.2 wt.,
1.3 eq.),
distilled 2,3-dibromopropene (3.71 wt., 1.3 eq.), and 48% hydrobromic acid
(0.002 vol.),
respectively. After observation of reaction initiation indicated by a
temperature spike to 36-
38 C, the second reactor was charged with 2,3-dibromopropene portion-wise (9 x
0.37 wt.)
while maintaining the internal temperature below 45 C. After complete
addition, the
contents of the second reactor were stirred at an internal temperature of 35 C
for an
additional 60 minutes. The filtered contents of the first reactor were charged
into the second
reactor at a rate such that internal temperature did not exceed 45 C. After
complete addition,
the heat source was removed and the second reactor was charged with Celite
545 (2.0 wt.)
and the resulting mixture stirred for 30 minutes. The heterogeneous mixture
was filtered
through a Celite 545 pad (2.0 wt.) and the cake washed with additional water
(5 vol.). All
filtrates were combined into a reactor and charged with concentrated
hydrochloric acid (1.5
vol.) until the cloudy solution becomes clear. With vigorous stirring, the
reactor was charged
with sodium chloride (3.6 wt.) and the layers allowed to partition. The
organic layer was
separated and set aside. The aqueous layer was extracted with n-butanol (20
vol.). The
aqueous layer was drained and the reactor charged with the organics from the
first separation.
The organics were washed with concentrated sodium bicarbonate (24 vol.),
followed by a
back extraction of the aqueous layer with n-butanol (20 vol.). All organics
were combined
and concentrated in vacuo. The concentrate was dissolved in MTBE (10 vol.),
filtered and
the filtrate was concentrated to two vol. With stirring, the concentrate was
cooled to an
internal temperature of 0 C and then n-heptane (4 vol.) was added. The
heterogenous mixture
was stirred for 2 hours at an internal temperature of 0 C and the desired
product was isolated
via filtration and dried under vacuum to yield ER-806909 (1.34 wt., 0.45 eq.)
as a white
powder.
OH TBDPSCI, imidazole OH
Br
OH Br OTBDPS
DMF, 0-15 C
ER-806909 ER-806545
[00106] A reactor was charged with imidazole (0.65 wt., 2 eq.), ER-806909
(1 wt., 1
eq.), and anhydrous DMF (4.04 vol.). With stirring, the reactor was cooled to
an internal
temperature of 0 C and then with teri-butylchlorodiphenylsilane (1.25 wt.,
0.95 eq.) at a rate
such that internal temperature did not exceed 15 C. While maintaining the
internal
42

CA 02822994 2013-08-01
temperature < 15 C, the reaction was stirred for an additional 1 hour. The
reactor was
charged with water (3.2 vol.) and n-heptane (6.4 vol.). The mixture was
stirred for 5-15
minutes and the layers allowed to separate. The organic layer was separated
and set aside.
The aqueous layer was extracted with n-heptane (3.2 vol.). All organics were
combined and
washed with brine (3.2 vol.) and concentrated in vacuo to a constant weight to
yield ER-
806545 (2.13 wt., 0.95 eq.) as a yellow oil. The product was utilized in the
next stage
without further purification.
SMB Separation
_OH
Br Br
OTBDPS ___________________________________ OTBDPS
Br
ER-806545 ER-808373 ER-806721
[00107] The enantiomers of ER-806545 were separated via Simulated Moving
Bed
(SMB) chromatography to yield ER-808373 (0.55 wt., 0.55 eq.) and ER-806721
(0.45 wt.,
0.45 eq.) as yellow oils. The SMB chromatography protocol used to separate the

enantiomers of ER-806545 is as follows.
Columns and media: Chiracel OD 20 gm 30 mm x 150 mm (12 columns)
Solvent system: 96:4 (vol/vol) heptane:tert-butanol (mobile phase)
Simulated Moving Bed
Chromatography apparatus: Knauer SMB System CSEP C912
Isotherms (Langmurian, determined by frontal analysis):
Undesired isomer: Qi* __ 2.8768 x Ci/(1+0.02327 x Ci)
Desired isomer: Qi=4.5320 x Ci/(1+0.0034595 x Ci) -
*Where Qi =solid phase concentration (in g/L) and Ci= liquid phase
concentration
(g/L).
Column porosity: 0.658
Temperature: 27 'V
Flow rates, etc. calculated by simulation on EuroChrom 2000 for Windows, SMB
Guide
ver.1.2, Wissenschaftliche Geratebau Dr.-Ing. Herbert Knauer GmbH, D-14163
Berlin;
authors, H. Kniep and A. Seidel-Morgenstern:
=
Feed concentration: 36 g/L (ER-806545)
Feed flow rate (Pump 1): 15 mL/min
Eluent flow rate (Pump 2): 76.4 mL/min
Zone IV (Pump 3) flow rate: 107 mL/min
Zone If (Pump 4) flow rate: 134.3 mL/min (actual flow rate = 143.5 mL/min)
Zone I flow rate: 183.4 mL/min
Zone III flow rate: 1493 mL/min
Raffinate* flow rate: 42.3 mL/min *Raffinate = weakly bound isomer
Extract flow rate: 49.1 mL.min *Extract strongly bound isomer
43

CA 02822994 2013-08-01
Tact time
(port switching time): 0.8864 min (53.18 sec, actual tact time = 54 sec)
[00108] The enantiomers of ER-806545 were separated using the above
protocol in the
following manner. For an 11 hour run, 10 L of 36 g/L ER-806545 in mobile phase
was
pumped (Silog model Chemtech) through a 142 mm diameter 0.45 i.tm pore size
nylon filter
(Cole-Parmer # 2916-48) and into the Feed tank. The Eluant tank was filled
with 36L mobile
phase that has been filtered through an in-line 45 mm diameter 1 pill glass
fiber filter
(Whatman GFC), additional vol. were added throughout the run. The internal
temperature of
the SMB apparatus was adjusted to 27 C.
[00109] For initial startup, the Feed and Eluant inlets were both connected
to the Eluant
tank. The Feed and Eluant pumps were primed and purged with mobile phase
solvent. The
SMB apparatus column switching was initiated, the pumps were turned on and the
flow rates
were gradually increased to full speed while maintaining absolute flow rate
differences
between each pump. Once full speed was achieved the Raffinate and Extract flow
rates were
measured and adjustments to pump flow rates were made to correct for
deviations in pump
specifications. The Feed pump (Pump 1) was reduced to 0 mL/min, the inlet
reconnected to
the Feed tank, the pump primed with Feed solution and then the flow rate
gradually
increased back to full operating speed. The Raffinate and Extract outlets were
collected into
separate tanks and samples of each were acquired every 2 hours. The samples
were
monitored for chiral purity by analytical HPLC using the HPLC method set forth
below.
Adjustments to the flow rates of Pumps 2, 3 and 4 as well as to the tact time
were made to
afford the desired outlet purities.
[00110] At the end of the run, the Feed pump was once again reduced to zero
flow rate
and connected to the Eluant tank. The Feed pump was brought back to full speed
and the
system was allowed to wash for 20 minutes. The Raffinate and Extract outlets
were
maintained for 10 minutes (10 tacts) during the wash period and, for the
remainder of the
wash, the outlets were collected into a separate tank. The column wash was
eventually
concentrated and added to the Feed on subsequent runs.
[00111] The collected Extract (ER-806721) at the end of each run was pooled
with
material collected from the same starting material lot and the final pooled
lot was analyzed
again for chiral purity by the analytical HPLC method described in Table 1
below. The
same procedure was applied to the collected Raffinate (ER-808373).
44

CA 02822994 2013-08-01
Table 1. HPLC Analysis of ER-806721 chiral purity:
Column: Chiracel OD 10 urn 250 x 4.6 mm, DAICEL Chemical Industries,
Ltd., cat. no. 14025
Flow rate: 0.8 mL/min
Temp. ( C): 27 preferred over 35
Inj. Vol.: 10 uL usually, sample in Solvent A, 5 mg/mL
Instrument: Waters Alliance W2690 with UV W2487 (also with Advanced Laser
Polarimeter)
Mobile Phase Constituents: (PDR-Chiral, Inc.)
A 99:1 Heptane 2-Propanol
Gradient Table: (%) Gradient
Time (min)
0 100 0 0 0 isocratic
Run time 30 min
Detection: Absorbance at 254 nm UV
PNBA, DEAD, PPh3,
Toluene, 0 C BrOPNB LION, THE, H20
r911
OTBDPS _____________________________ OTBDPS
Br Br
ER-808373 ER-806721
[00112] A reactor was charged with triphenylphosphine (0.7 wt., 1.2 eq.), p-
nitrobenzoic
acid (0.45 wt., 1.2 eq.), ER-808373 (1 wt., 1 eq.), and anhydrous toluene (8
vol.). The
reaction was cooled to internal temperature of 0 C and DEAD (1.17 wt., 1.2
eq.) was
slowly added at a rate such that the internal temperature did not exceed 7 C.
n-Heptane (3.3
vol.) was added and the mixture cooled to an internal temperature 10 C then
stirred for 30-
40 minutes. The resulting precipitate was removed by filtration. The filter-
cake was washed
with n-heptane (3.3 vol.), TBME (0.55 vol.), n-heptane (1.1 Vol.), and MTBE
(0.55 vol.),
respectively. All filtrates were combined and concentrated in vacuo. The crude
concentrate
was dissolved in THF (8 vol.) then water (0.8 vol.) and lithium hydroxide
dihydrate (0.18
wt., 2 eq.) were added. The mixture was stirred at ambient temperature then n-
heptane (3.3
vol.) was added and stirred for 5 minutes. Water (2.2 vol.) and n-heptane (3.3
vol.) were
added, the biphasic mixture was stirred for 5 minutes, and the layers were
allowed to
partition. The

CA 02822994 2013-08-01
aqueous layer was separated and back extracted with n-heptane as necessary.
The organic
layers were combined and concentrated in vacuo. The crude product was purified
via Si02
column chromatography to yield ER-806721 (0.74-0.85 wt., 0.74-0.85 eq.) as a
light yellow
oil.
OH TsCI, DMAP, CH2Cl2. OTs
it, 48 hr
O
Br TBDPS BrOTPS
ER-806721
ER-807204
[00113] A reactor was charged with ER-806721 (1 wt., 1 eq.) and anhydrous
dichloromethane (4.2 vol.). The reaction was cooled to an internal temperature
of 0-5 C,
then triethylamine (0.34 wt., 1.5 eq.), p-toluenesulfonyl chloride (0.51 wt.,
1.2 eq.), and 4-
(dimethylamino)-pyridine (0.001 wt., 0.25 eq.) were added. The resulting
mixture was
stirred at ambient temperature for 48 hours then water (1.8 vol.) and
dichloromethane ( 1.8
vol.) were added. After sufficient mixing, the organics were separated and
concentrated.
The concentrate was dissolved in MTBE (1.8 vol.) and washed with brine (1.8
vol.). The
organic layer was separated and set aside. The aqueous layer was back
extracted with
MTBE (1.8 vol.) then all organics were combined and concentrated in vacuo. The
crude oil
was filtered through a plug of Si02 (70-230 mesh, 1 wt.) eluting with MTBE (7
vol.) and the
filtrates were concentrated in vacuo. The concentrate was dissolved in IPA (5
vol.) and
water (0.25 vol.) was added. The resulting mixture was cooled to an internal
temperature of
15 C and then seeded with ER-807204. After seeding, the mixture was cooled to
an
internal temperature of 0 C and stirred for 4-5 hours. The suspension
filtered, the filter cake
was washed with cold IPA (1 vol.), and the cake dried in vacuo to a constant
weight to yield
ER-807204 (1.05 wt., 0.78 eq.) as a white powder. IR (thin film, cm-1) X 2597,
1633, 1363,
1177, 907, 729. LRMS link 602 (M + H).
1) diethylmalonate, 21% Na0Et in Et0H, 65 C 0
0õ 0
2) MgC12, DMF, .125 C
ER-806906 ER-805552
[00114] A reactor was charged with 21% sodium ethoxide in ethanol (2.97
wt., 0.9 eq.).
The solution was heated to an internal temperature of 65 C then diethyl
malonate (3.24 wt.,
2 eq.) was added at a rate such that the internal temperature did not exceed
70 C. The
mixture was stirred for 30 minutes and then ER-806906 (1 wt., 1 eq.) was added
over 3-5
hours. Upon complete addition, the reaction was stirred for 60 minutes and
then cooled to an
46

CA 02822994 2013-08-01
internal temperature of 50 C. Concentrated hydrochloric acid (0.84 wt., 1.05
eq.) was added
at a rate such that internal temperature did not exceed 65 C. The DMF (3 vol.)
and ethanol
were removed via distillation then a solution of magnesium chloride
hexahydrate (0.21 wt.,
0.1 eq.) in distilled water (0.25 vol., 1.4 eq.) was added. The resulting
mixture was heated to
an internal temperature of 135 C while removing the distillate. The mixture
was heated at
reflux then cooled to room temperature and brine (12 vol.) and TBME (16 vol.)
were added.
The organic layer was separated and washed with water (1.3 vol.) and brine
(1.2 vol.) then
concentrated in vacuo. The product was purified via distillation to yield ER-
805552 (0.95-
1.09 wt., 0.71 eq.).
0 0
0 0
Methyl iodide, LiHMDS in toluene,
ER-805552 ER-806724
THF, -78 C
[00115] A reactor was charged with LHMDS 1.0 M in toluene (6.61 wt., 1.04
eq.) and
cooled to an internal temperature of -75 'C. ER-805552 (1 wt., 1 eq.) was
dissolved in
anhydrous THE and added to the reactor at a rate such that internal
temperature did not
exceed -70 C. Upon complete addition, the resulting mixture was stirred for 30
minutes. A
second reactor was charged with anhydrous THF (2.5 vol.) and methyl iodide
(1.27 wt., 1.25
eq.) and cooled to an internal temperature of -75 C. The solution of ER-805552
in THE was
added into the methyl iodide solution at a rate such that internal temperature
did not exceed
¨65 C. Upon complete addition, the reaction was stirred at an internal
temperature of -78 C
for 30 minutes. The reaction was inverse quenched a solution of! N
hydrochloric acid (10
vol.) and MTBE (8 vol.) with vigorous stirring_ After complete addition, the
aqueous layer
was separated and discarded. The organic layer was washed with brine solution
(3 vol.) and
concentrated in vacuo and the product purified via distillation to afford ER-
806724 (0.75
wt.) as a ¨6 /1 mixture of diastereomers.
0 o
1) dimethylhydroxylamine, AlMe3 (2M in toluene) 0
CH2Cl2, 0 C
OTBS
2) TBS-CI, imidazole, DMF, it
ER-806724 ER-806753
C9H1402 C17H35NO3Si
Mot_ Wt.: 15421 Mol. Wt: 329.55
6.5 / 1 anti / syn
47

CA 02822994 2013-08-01
[001161 A reactor was charged with N,0-dimethylhydroxylamine 1-IC1 (1.05
wt., 1.5 eq.)
and anhydrous CI-12C12 (8.1 vol.) and cooled to an internal temperature of 0
C. 2 M
trimethylaluminum in toluene (3.93 wt., 1.5 eq.) was added at a rate such that
internal
temperature did not exceed 5 C. The reaction was stirred for an additional 10
minutes and
ER-806724 was added at a rate such that internal temperature did not exceed 5
C. The
reaction was diluted with CH2C12 (15 vol.) then inverse quenched into 1.3 M
sodium tartrate
(20 vol.) at an internal temperature of 0 C at a rate such that the internal
temperature did not
exceed 10 C. After complete addition, the layers were partitioned and the
aqueous layer
was separated and set aside. The organics were washed with water (1 vol.),
dried over
sodium sulfate (1 wt.), filtered and concentrated in vacuo until minimal
methylene chloride
was being removed. To the concentrate was added anhydrous DIVEF (6.3 vol.),
imidazole
(0.64 wt, 1.5 eq.) and t-butyldimethylsilyl chloride (0.94 wt., 0.97 eq.),
respectively. Water
(5 vol.) and MTBE (10 vol.) were added and the resulting mixture stirred then
the layers
were allowed to partition. The aqueous layer was separated and discarded. The
organic
layer was washed with water (5 vol.) and the layers separated. 1 N sodium
hydroxide (2.5
vol.) and methanol (2.5 vol.) were added and the resulting mixture stirred.
The aqueous
layer was separated and the organic layer washed with brine (2.5 vol.) then
concentrated in
vacuo to afford ER-806753 (1.94 wt., 0.91 eq.) as a brown oil.
0 0
i) 0s04, NMO, CH2Cl2
OTBS OTBS
ii) Na104, Na104, THF
ER-806753 phosphate buffer pH=7 ER-806754
[00117] A reactor was charged with ER-806753 (1 wt., 1 eq.), CH2C12 (5
vol.) and
NMO-50% in water (0.8 wt., 1.1 eq.). The mixture was cooled to an internal
temperature of
C and then 0.197 M 0s04 in toluene (0.06 vol., 0.004 eq.) was added. Sodium
sulfite
(0.1 wt., 0.25 eq.) and water (0.85 vol.) were added and the reaction stirred
for 1 hour. The
mixture was diluted with brine (0.85 vol.) and the organics were concentrated
in vacuo to
approximately 1/3 vol. A second reactor was charged with sodium periodate (1.3
wt., 2 eq.)
followed by THF (2.5 vol.). The mixture was diluted with p1-1 = 7 phosphate
buffer (3.0
vol.) and cooled to an internal temperature of 20 C. The concentrated diol was
added at a
rate such that the internal temperature did not exceed 30 C. After complete
addition, the
resulting mixture was stirred at room temperature. Water (1.25 vol.), MTBE (7
vol.) and
brine solution
48

CA 02822994 2013-08-01
(1.25 vol.) were added and the layers separated. The organics were washed a
second time
with a mixture of brine solution (1 vol.) and saturated sodium bicarbonate (1
vol.). Finally,
the organics were stirred over a mixture of brine (1 vol.) and 10% (w/v)
sodium thiosulfate
solution (1 vol.) for 1 hour then concentrated in vacuo. The crude material
was purified via
Si02 column chromatography to yield ER-806754 (0.93 wt., 0.93 eq.) as a yellow
oil. IR
(thin film, cm-1) 2953, 2856, 1725, 1664, .1463, 1254, 1092, 833. LRMS m/z 332
(M + H).
I Me OTBS
o
0 OP
0 Me
0"N
NHMs TBSO o
I 0
ER-806754
ER-806629 Me
(3.1 eq)
,õOH
(R)-Ligand OTs
OTBDPS
1. CrCl2 (3.1 eq),
TEA (3.1eq),
OTBDPS =
Br NiC12( m eq),
THF (30 mUg),
25 C, 3 h
ER-807204 2. H2N(CH2)2t4H2
TBSO
Me
IPA
"
OTBDPS
Si02
17 14
25 c
ER-807524
[00118] A reactor was charged with ER-806629 (1.53 wt., 3.1 eq.) and TI-IF
(10.5 vol.)
and the solution was degassed with nitrogen sparge for 60 minutes. A second
inerted reactor
was charged with ER-807204 (1 wt., 1.0 eq.), ER-806754 (0.66 wt., 1.2 eq.) and
TI-1F (2.7
vol.) and this solution was degassed with argon sparge for 45 minutes. The
reactor
containing ER-806629 was charged with CrCl2 (0_63 wt., 3.1 eq.) and followed
by Et3N
(0.52 wt., 3.1 eq.). The dark green suspension was stirred at an internal
temperature of 30 to
35 C for 1 hour, cooled to 0 to5 C and then NiCl2 (0.1 eq.) was added. The
first reactor
was charged with the contents of the second reactor slowly over 0.5 hours and
the reaction
allowed to warm to rt. The reaction was cooled to an internal temperature of 0
C then
ethylenediamine (1.0 wt., 10 eq.) was added over 30 minutes and the reaction
stirred at an
internal temperature of 25 C for at least 30 minutes. To the reaction was
added water (4
vol.), TBME (10 vol.) and n-heptane (1 vol.) and the resulting mixture stirred
for 15 minutes
and the phases allowed to separate (-30 min). The aqueous phase was separated
and back
extracted with TBME (-7.5
49

CA 02822994 2013-08-01
vol.). The organic layers were combined and washed with water (5 vol.), brine
(3 vol.), and
concentrated in vacuo to minimum volume. To the crude mixture was added IPA
(10 vol.)
and Si02 (1 wt.) and the resulting mixture stirred at an internal temperature
of 25 C for up to
4 days. The slurry was filtered and the filter cake washed with IPA (2 X 1
volume). To the
filtrate was added n-heptane (6.6 vol.) and the mixture was concentrated in
vacuo until a
suspension fointed. The mixture was filtered and the cake washed with n-
heptane then the
mixture was concentrated in vacuo. The crude product was purified via Si02
column
chromatography to yield ER-807524 (0.54 wt., 0.48 eq.), as a clear yellow oil.
IR (thin film,
cm-1) X 2934, 1668, 1471, 1108, 833. LRMS m/z 704 (M + Na).
o'
TBSO 0 N TBSO ' 0,...õ---
"-
-
MeMgCI
_____________________________________ ).- 0
0 THF, -20 C ''',5 t-----7-''OTBDPS
OTBDPS
ER-807524 ER-807525
[00119] A reactor was charged with ER-807524 (1 wt., 1 eq.) and anhydrous
THE (1.25
vol.). The mixture was cooled to an internal temperature of -20 C and 3 M
methyl
magnesium chloride (0.59 vol., 1.2 eq.) was added at a rate such that the
internal
temperature did not exceed 0 C. Upon complete addition, the mixture was warmed
to an
internal temperature of 0 C over 2 hours. The reaction mixture was inverse
quenched into
semi-saturated ammonium chloride (2.62 vol.) and the resulting mixture diluted
with TBME
(2 vol.) with vigorous mixing. The aqueous layer was discarded and the
organics washed
with brine (2 vol.) then concentrated in vacuo. The crude product was purified
via Si02
column chromatography to yield ER-807525 (0.79-0.82 wt., 0.85-0.88 eq.) as
yellow oil.
0
TfO,
TBSO i) KHMDS, -78 C TBSO
THF /c/-',-
________________________________________ )=
0 0
2
1
OTBDPS ii) TfNPh, -78 to -20 C "-"-5 ---"---OTBDPS
ER-807525 ER-807526
[00120] A reactor was charged with ER-807525 (1 wt., 1 eq.), N-
phenylbistrifluommethanesulfonamide (0.59 wt., 1.1 eq.), and anhydrous THF
(4.1 vol.) and
the mixture cooled to an internal temperature of -75 C. 0.5 M KHMDS in toluene
(2.75 wt.,

CA 02822994 2013-08-01
1 eq.) was added at a rate such that internal temperature did not exceed -60 C
then the
reaction was warmed to-20 C over 2 hours. The reaction was quenched with semi-
saturated
NI-WI (2.4 vol.) at a rate such that internal temperature did not exceed 0 C.
The mixture was
warmed to an internal temperature of 20 C and n-heptane (2.4 vol.) was added.
The mixture
was stirred and the aqueous layer was separated and discarded. The organic
layer was
washed three times with saturated sodium bicarbonate (2.3 vol. each) then
concentrated in
vacuo to yield ER-807526 (1.2-1.4 wt., 1.0-1.2 eq.). The material was utilized
in the next
stage without further purification.
TfO,
TBSO HOTf0\.
HCI in IPA, Me0H
OTBDPS 0
loOH
ER-807526 ER-807527
[00121] A reactor was charged with ER-807526 (1 wt., 1 eq.) and anhydrous
methanol (3
vol.) at 20 C. 1.25M HCI in IPA (4 vol., 5 eq.) was added and the mixture
stirred for 3
hours. Solid NaHCO3 (0.42 wt., 5 eq.) was added portion-wise with stirring
until the pH of
the reaction mixture reached 6-7. The reaction mixture was filtered with
methanol (3 x 2
vol.) washes. All filtrates were concentrated in vacuo and then purified via
Si02 column
chromatography to yield ER-807527 (0.43 wt., 0.79-0.85 eq.).
OH
HPLC
_____________________________________ )1r
Separation
I 0H nOH
ER-807527 ER-806730
[00122] The diastereomeric mixture of ER-807527 was separated by
preparative HPLC
chromatography and the desired fractions concentrated to yield ER-806730 (0.56
wt., 0.56
eq.) as a clear yellow oil. The preparative HPLC chromatography protocol used
to isolate
ER-806730 is as follows.
Column and Media: Kromasil spherical silica 60 A, 10 pill particle size
packed to
7.7 cm (diam.) x 30 cm (length) in a 7.7 cm x 60 cm Varian
Dynamax Rampak column.
HPLC Packing Station: Varian (Rainin) Dynamax Rampak 41/77 mm Column Packing
Station
51

CA 02822994 2013-08-01
HPLC Pumps: Varian (Rainin) SD-1 Titanium Pump Heads
Primary HPLC Detector: Waters R403 Refractive Index detector
Secondary HPLC Detector: Varian (Rainin) UV-1 detector with preparative flow
cell
Chromatography Control
and Acquisition Software:Varian (Rainin) Dynamax DA version 1.4.6
Chromatography Data
Processing Software: Varian (Rainin) Dynamax R version 1.4.3
Mobile Phase: 28.5 : 63.7 : 7.85 (vol/vol) n-heptane: methyl tert-
butyl ether:
2-propanol
Flow Rate: 140 mL/min
Column Temperature: Ambient (25 C)
Detection: Refractive Index, negative polarity at 16 X attenuation
and UV
at 215 nm.
Mobile Phase Gradient: Isocratic
Run Time: 40 minutes
Injection Volume: 10 ml of 0.8 g/mL of ER-807527
[00123] The above protocol was used to separate the diastereomers of ER-
807527 in the
following manner. Each lot of ER-807527 was first diluted to 0.1 g/m1 in the
mobile phase
and filtered under vacuum through a 47 mm, 1 gm pore size, glass fiber filter
(Whatman
GFC). The filtrate was then concentrated under vacuum on a rotary evaporator.
Flow on the
SD-1 HPLC pump A (primed and purged with mobile phase) was initiated and the
flow rate
gradually increased to 140 mL/minute. The system was washed until the UV and
RI
detectors achieved a stable baseline. The RI detector reference flow cell was
flushed with
fresh mobile phase.
[00124] Chromatography of 8 g injections of ER-807527 was accomplished by
diluting
the current lot of ER-807527 to a concentration of 0.8 g/mL in the mobile
phase. Injecting
mL aliquots of the dissolved material and collecting the eluant corresponding
to the ER-
806730 peak approximately beginning at the peak apex approximately 24 minutes
and
continuing to 35 minutes. Subsequent injections and fraction collection were
continued until
the starting material is exhausted.
[00125] The fractions corresponding to ER-806730 were pooled and
concentrated under
vacuum on a rotary evaporator. The diastereomeric purity and area-% purity
area were
assessed using the HPLC analytical method described in Table 2.
52

CA 02822994 2013-08-01
Table 2. HPLC Analysis of Diastereomeric Purity of ER-806730:
Column: Kromasil Slica 250 x 4.6 mm, 5 tun , MetaChem cat. no. 0475-250X046
Flow rate: 1 mL/min
Temp. ( C): 27
Inj. Vol.: 10 uL, sample in Solvent A, 2.5 mg/mL
Instrument: Waters Alliance W2690 with UV W2487
Mobile Phase Constituents:
A 30:67:3 n-Heptane:Methyl tert-Butyl Ether:2-Propanol
2-Propanol
Gradient Table: (oh) Gradient
Time (min) A
0 100 0 0 0 isocratic
22 100 0 0 0 isocratic
26 90 10 0 0 linear
32 90 10 0 0 isocratic
Run time 32 min with 18 min re-equilibration time at initial conditions
Detection: Absorbance at 205 nm UV
Tf0 Tf.0
HO HO s---
)1\/".õ PvCI, collidine, DMAP
0 0
rOH
DCM, 0 C .0Pv
ER-806730 ER-806732
[00126] A reactor was charged with ER-806730 (1 wt., 1 eq.) and anhydrous
dichloromethane (4.8 vol.) and cooled to an internal temperature of 0 C. 2,4,6-
Collidine
(1.16 wt., 4 eq.) and DMAP (0.03 wt., 0.1 eq.) were added and the resulting
mixture stirred
for 15 minutes and then trimethylacetyl chloride (0.3 wt., 1.05 eq.) was added
at a rate such
that internal temperature did not exceed 10 C. Water (3 vol.) was added and
the mixture
stirred for 15 minutes. TBME (10 vol.) was added and the mixture stirred for
an additional
minutes. The organic layer was washed with IN HC1 (10 vol.) washing until a
negative
result for 2,4,6-collidine is obtained then with water (5 vol.), saturated
sodium bicarbonate
(5 vol.), and saturated brine (5 vol.), respectively. The organic layer was
concentrated in
vacuo and the concentrate purified via Si02 column chromatography to yield ER-
806732
(1.02 wt., 0.85 eq.) as a yellow oil.
53

CA 02822994 2013-08-01
Tf0 TIO
HO Ms0
MsCI, EI3N, THF
_______________________________________ ). 0
OPv
'OPv
ER-806732 ER-805973
[00127] A reactor was charged with ER-806732 (1 wt., 1 eq.) and anhydrous
THF (2.35
- vol.) and cooled to an internal temperature of 0 C. Triethylamine (0.22
wt., 1.1 eq.) was
added followed by methanesulfonyl chloride (0.24 wt., 1.05 eq.) at a rate such
that internal
temperature did not exceed 10 C. The reaction was stirred at an internal
temperature of 0 C
then n-heptane (3.4 vol.) was added with vigorous stirring and the layers were
allowed to
partition. The organics were washed with saturated brine (3.4 vol.), dried
over saturated
sodium sulfate (2 wt.), filtered and the cake washed with n-heptane until a
negative result for
ER-805973 (F-2a) was obtained. The filtrates were concentrated in vacuo to
obtain ER-
805973 (1.12 wt., 0.97 eq.). The crude ER-805973 (F-2a) was used in the next
stage
without further purification. IR (thin film, cm-1) X 2961, 1725, 1413, 1208,
926. LRMS m/z
579 (M + H).
Example 3
Alternate Preparation of ER-806730:
HO2C 0 .H
OH 1. Cyclohexanone, H2SO4
-.0
2. NaHCO3, Toluene HO"'
OH
quinic acid 1
[00128] Quinic acid (lwt), cyclohexanone (2.11 eq, 1.08 wt), and conc.
sulfuric acid
(0.011 eq, 0.0056 wt) were added to a reactor. The reaction mixture was heated
to 160 C
and water was removed by azeotropic distillation (azeotrope begins at 100 C).
The reaction
was cooled to 90 C to 100 C and sodium bicarbonate (0.0096 wt) and toluene
(3.6 wt)
were added. The reaction was cooled to ambient temperature over 4-6 hours and
the
resulting precipitate was filtered, washed with toluene (2 x 0.9 wt), and
dried to provide 1
(0.97 wt) as a white powder.
54

CA 02822994 2013-08-01
H
1. TMSCI, imid. THF
0
-.0 2. DIBAL-H RO's\il
HO's\-1 3. a. AcOH, H20
"0
b. Et3N, DMAP
Ac20
1 C. recrystallized 2a, X=0, R=TMS
2b, X=H, OH, R=TMS
2c, X=F1, OAc, R=Ac
[00129] Compound 1(1 eq, 1 wt) and imidazole (2.5 eq, 0.80wt) were
combined, purged
with N2, and supended in anhydrous THF (10 v). TMSC1 (1.2 eq, 0.61 wt) was
added at a
rate that maintained the temperature below 30 C. The reaction was cooled to
ambient
temperature, heptane (10 v) was added and the resulting suspension filtered.
The filter cake
was washed with 1:1 hpt/THF (10 v) the filtrate solvent was exchanged with
toluene by
atmospheric distillation to provide a solution of 2a (calcd. at 1.34 wt) in
toluene (-5 wt).
[00130] The solution of 2a was cooled to ¨78 C and DIBAL-H (L5 M in
toluene, 1.2
eq, 2.1 wt) was added at a rate that maintained the temperature below ¨65 C.
The excess
DIBAL-H was quenched with Me0H (0.3 eq, 0.034 wt) and the solution warmed to 0
C.
The solution was transferred to a solution of 30% wt/wt aqueous Rochelle salt
(10 wt) and
sodium bicarbonate (1 wt) at a rate that maintained the temperature below 25
C. The
mixture was stirred vigorously to obtain a biphasic solution. The layers were
separated and
the aqueous layer was extracted with MTBE (5v). The combined organic layers
were
washed with water (2.5 wt) and then saturated brine (2.5 wt). The organics
were
concentrated and solvent exchanged with THF to provide a solution of 2b
(calcd. at 0.98 wt)
in THF (5 v).
[00131] The solution of 2b was cooled to 5 C and acetic acid (2.9 eq, 0.51
v) was added.
Water (LO eq, 0.055 v) was added and the solution stirred at 0 C to 5 'C. Up
to two
additional aliquots of acetic acid and one aliquot of water were added as
needed to facilitate
deprotection of the say] group. Et3N (12 eq, 3.6 wt) and DMAP (0.05 eq., 0.02
wt) were
added at a rate that maintained the temperature below 20 C. Acetic anhydride
(6 eq, 2.0 wt)
was added and the reaction stirred at rt. The reaction was cooled to 5 C and
added to
saturated aqueous sodium bicarbonate (10 v) at a rate that maintained the
temperature below
30 C. The resulting mixture was allowed to stir for 3-4 hours and the layers
allowed to
separate. The aqueous layer was extracted with MTBE (5 v) and the combined
organics
were washed with water (5 v). The extracts were solvent exchanged with IPA by
distillation
to provide a solution of 2c in IPA (3 v). The solution was cooled to 5 C and
the resulting

CA 02822994 2013-08-01
crystals were filtered. The mother liquor was concentrated and a second crop
obtained after
recrystallization to provide 2c (0.87 wt) as a white crystalline solid.
H
TMS Me02C 0
0 , õH
AcO5
MeO2CJ.
-0 AcO"'
AcOv
BF3-0Et2, ACN c0-10
TFAA
2c 3
[00132] 2c (1 wt) was dissolved in acetonitrile (6 v) and methyl 3-
trimethylsilylpent-4-
eneoate (3.0 eq, 1.86 wt) was added followed by TFAA (0.2 eq, 0.083 v). BF3-
0Et2 (1.0 eq,
0.42 v) was then added to the solution at a rate that maintained the
temperature below 25 C.
The reaction was added to saturated aqueous sodium bicarbonate (10 v) and the
resulting
mixture stirred for 15 minutes. The mixture was extracted with heptane (10 v),
followed by
MTBE (5 v) and the combined extracts were concentrated to provide 3 (calcd. at
0.72 wt) as
an orange oil.
Me02C 0
Na0Me, Me0H Me02C
AcO"s 0
4
3
[00133] A solution of 3 (1 wt) in THF (9 v) was treated with sodium
methoxide (25%
wt/wt in methanol (1.5 eq, 2.2 wt)) at a rate that maintains the temperature
below 25 C. The
reaction was quenched by addition to 1 N HC1 (10 v). The organic layer was
separated and
the aqueous was extracted with MTBE (10 v). The combined organics were washed
with
water (2.5 v), saturated sodium bicarbonate (2.5 v), and water (2.5 v). The
solution was
concentrated to provide a solution of 4 (calcd. at 0.88 wt) in THF (2.5 v).
The solution was
used directly in the next step.
0 0
õH 1N HCI os
Me02C Me02C
0 111111V,0 Me0H, THF 0
bH
4 5
[00134] Methanol (5 v) was added to the solution of 4 (1 wt) in
tetrahydrofuran (2.5 v).
IN HCI (0.75 eq, 2 v) was added and the reaction was warmed to 60-80 C. The
reaction
was cooled to rt and added to saturated aqueous bicarbonate. The mixture was
extracted
with
56

CA 02822994 2013-08-01
DCM (3 x 2.5 v) and the combined DCM extracts were solvent exchanged with
Et0Ac to
provide a solution of 5 in Et0Ac (3 y). 1-leptane (2 v) was added to induce
crystallization
and the resulting suspension cooled to 0 C. The solids were collected by
filtration and the
filter cake washed with cold Et0Actheptane (1:1 v/v) and dried to provide 5
(0.55 wt) as a
white powder.
H
0 1) 2-acetoxy-2-
methylpropanyi bromide 0
H20, ACN
Me02CMe02C
oIWO "'OH 0
2) Na0Me, Me0H
OH 6
[001351 Compound 5 (1 wt) was dissolved in ACN (10 v) then 2-acetoxy-2-
methylpropanyl bromide (4.0 eq, 2.2 wt) and water (1 eq, 0.067 wt) were added
consecutively. The resulting mixture was stirred at ambient temperature then
cooled to 5-10
C. NaOMe (25% wt/wt in Me0H, 8 eq, 6.2 wt) was added and the reaction allowed
to
warm to ambient temperature. The reaction was quenched by the addition of
saturated
sodium bicarbonate (10 v) and extracted with MTBE (2 x 10 v). The solvent was
exchanged
with methanol by atmospheric distillation to provide a solution of 6 (calcd at
0.91 wt) in
methanol (10 v).
0
0 NaBH4, Me0H, 55 C
Me02C 4- NO 0 *V
.=.b
7
6
[001361 A solution of 6 (1 wt) in methanol (10 v) was heated to,55 C.
Sodium
borohydride (5 eq, 0.68 wt) was added in 6 portions and the reaction cooled to
5 C and
quenched with 1 N HC1 (10 v). Brine (5 v) was added and the reaction extracted
with
Et0Ac (2 x 10 v). The extracts were combined and concentrated to provide 7 as
a tan
residue.
0 õH
TBDPSCI, imid
HO 0 4-
TBDPSO 0
..,b
8
7
[001371 Compound 7 (1 wt) was dissolved in CH2C12 (10 v) then DMAP (0.1 eq,
0.054
wt), Et3N (3.0 eq, 1.85 v), and 1BDPSCI (1.2 eq, 1.38 v) were added at ambient

temperature. Sodium bicarbonate (10 v) was added and the organic layer
separated. The
57

CA 02822994 2013-08-01
aqueous layer was extracted again with CH2Cl2 (10 v), the organic extracts
combined and
concentrated to provide 8 (calculated at 1.8 wt) as a colorless oil.
0
0 sõH
LDA, THE, 50 C
TBDPSO 0 11 TBDPSO 0 .,,OH
9
8
[00138] LDA (1.5 M in cyclohexane, 4 eq, 6 v) was added to a solution of 8
(1 wt) in
THF (10 v) at ambient temperature. The solution was warmed to 50 C then
quenched with
1 N HC1 (5 v) and extracted with MTBE (10 v). The extracts were concentrated
to provide 9
(0.9 wt) as an oil.
OH
0 H = OH
,
1) 03, CH2Cl2, IVIe0H
TBDPSO 0 Vt. '"OH0
TBDPSO
2) NaBH4 OH
9 10
[00139] A solution of 9 (1 wt) in CH2C12 (5 v) in Me0H (5 v) was cooled to
¨60 C and
treated with 03 keeping temperature below ¨50 C. The reaction was purged with
N2,
NaBH4 (0.5 eq, 0.04 wt) was added and the mixture warmed to 0 C. Additional
NaBH4 (1
eq, 0.08 wt) was added in portions and the reaction allowed to warm to ambient
temperature.
After 3 hours, the mixture was quenched with IN HCI,(10 v), CH2Cl2 (5 v) was
added and
the layers were allowed to partition. The aqueous layer was re-extracted with
CH2Cl2 (10 v)
and the organic extracts were combined and concentrated to provide 10 (0.97
wt) as a
colorless oil.
OH
H Na104, THE. H2O ,
TBDPS00 TBDPSO 0
OH 0
11
[00140] Compound 10 was dissolved in THF (10 v) and phosphate buffer (pH=7,
5 v)
was added. NaI04 (2 eq, 0.854 wt) was added and the reaction warmed to ambient

temperature. Water (5 v) and MTBE (10 v) were added and the resulting mixture
was stirred
vigorously for 10 minutes. The organic layer was separated and washed with 10%
wt/v
aqueous sodium thiosulfate (5 v), water (5 v), and brine (5 v) then dried by
azeotropic
distillation with THF (-200 ppm water) to provide a solution of 11 (calcd. at
0.93 wt) in
THF (10 v). This solution was used directly in next step.
58

CA 02822994 2013-08-01
0
PPh3CH2CO2Me
TBDPS00 THF 65 TBDPS00" \OH CO2Me
0 C
11 12
[00141] (Carbomethoxymethylene)triphenylphosphorane (1 wt) was added to the
solution of 11(1 wt) in THF (10 v) and heated to 65 C. Heptane (40 v) was
added and the
resulting mixture stirred for 30 minutes. The resulting precipitate was
filtered and the
filtrate concentrated to a total 10 v. Si02 (5 wt) was added and the
suspension filtered over a
pad of Si02 eluting with MTBE (20-40 v). The solvent was exchanged with Me0H
to
provide a solution of 12 (calcd. at 0.95 wt) in Me0H (10 v) which was used
directly in the
next step.
H2 Pd Me0H
_________________________________________ TBDPS00.-/\---\¨H CO2Me
OH O
12 13
[00142] A solution of 12 (1 wt) in Me0H (10 v) was added to 10% wt/wt Pd(C)
(0.23 eq,
0.37 wt) and treated with H2. The suspension was filtered while rinsing the
filter cake with
THF (10 v). The solvent was exchanged with THF to provide a solution of 13
(calcd. at 0.95
wt) in THF (10 v) which was used directly in the next step.
LAH, THF, 0 C
TBDPS00-- OH TBDPSOO V)----\¨0O2Me
OH \OH
13 14
[00143] The solution of 13 (1 wt) in THF (10 v) was cooled to 0-5 C and
LAH (1 M
(THF), 0.78 eq, 1.5 v) was added at a rate that maintained the temperature
below 10 C.
Water (1.7 eq, 0.06 v) was then added at a rate that maintained the
temperature below 10 C.
NaOH (10% wt/wt in water, 0.16 eq, 0.06 v) was added followed by water (4.98
eq, 0.17 v)
at a rate that maintained the temperature below 10 C and the resulting mixture
stirred
vigorously while warming to ambient temperature. The suspension was filtered
and the
filter cake rinsed with THF (5 v). The filtrate was partially concentrated to
provide 14
(calcd. at 0.9 wt) in THF (10 v) which was used directly in next step.
59

CA 02822994 2013-08-01
TrCI, THF, imid
TBDPSO
TBDPS00 \
OH OH OH OTr
14 15
[00144] The solution of 14 (1 wt) in THF (10 v) was cooled to 0 C then
imidazole and
TrC1 (1.5 eq, 0.59 wt) were added. Saturated aqueous NaHCO3 (5 v) was added
and the
mixture extracted with heptane (10 v). The extract was washed with brine (10
v) and
concentrated to provide 15 (1.35 wt).
TBAF, THF
TBDPS00HOO
OHOTr OH OTr
15 16
[00145] Compound 15 (1 wt) was dissolved in THF (10 v) and treated with
TBAF (1M,
1.2 eq, 1.6 v). The reaction mixture was concentrated to 2 v then heptane (5
v) and Si02 (5
wt) were added. The resulting suspension was filtered and eluted with heptane
(5 v)
followed by THF (10 v). The THF eluent was collected to provide a solution of
16 (calcd at
0.61 wt) in THF (10 v) which was used directly in the next step.
NIS, PPh3, py, THF
HOO
OH OTr OTr
16 17
[00146] PPh3 (5 eq, 2.3 wt), pyridine (10 eq 1 vol), and NIS (3.0 eq, 1.1
wt) were added
to the solution of 16 (1 wt) in THF (10 v). 20 % wt/wt aqueous citric acid (10
eq, 14 wt) was
then added and the resulting mixture allowed to stir for 10 minutes. The
reaction was
diluted with heptane (10 v) and the aqueous layer separated. The organic layer
was washed
with water (5 v), 10 % wt/v aqueous sodium thiosulfate (5 v), water (5 v) and
brine (5 v).
The solvent was exchanged with Et0H and concentrated to 5 v. Water (10 v) was
added and
the resulting precipitate was collected by filtration to obtain 17 (0.65 wt)
as a white solid.
KCN, Et0H, H20
0NCO
OTr 80 C OTr
17 18
[00147] Compound 17 (1 wt) and KCN (6 eq, 0.54 wt) were suspended in EtOFI
(5 v)
and water (10 v) and the resulting suspension heated to 80 C. The reaction
was diluted with

CA 02822994 2013-08-01
water (5 v) and Et0Ac (10 v) and mixed for 10 minutes. The aqueous layer was
removed
and the organic layer washed with water (5 v) and brine (5 v). The solvent was
exchanged
with Et0H to provide 18 (0.75 wt) in Et0H (10 v) which was used directly in
next step.
Zn, Et0H, 80 C
0
18 \OTr
n \OTr
19
[001481 Zn (37 eq, 3.9 wt) was added to the solution of 18 (1 wt) in Et0H
(10 v) and the
mixture heated to 75-80 C. The reaction was partially concentrated to 2-3 v,
cooled to
ambient temperature, and partitioned between MTBE (10 v) and water 5 (v). The
aqueous
layer was removed and the organic layer washed with saturated bicarbonate (5
v), water (5
v), and brine (5 v), then dried by THF azeotropic distillation to ¨200 ppm
water to provide
19 (0.81 wt) in THF (10 v). The resulting solution was used directly in next
step.
0 0
0
1. LDA, THF, -65 C
o 2. Mel
OTr
19 20
[001491 LDA (1.0 M in THF, 1.2 eq, 2.4 v) was added to the solution of 19(1
wt) in
THF (10 v) at ¨78 C. The resulting mixture was stirred for 10 minutes then
the enolate
solution was added to a solution of Mel (1.5 eq, 0.19 v) in THF (5 v) at ¨78
C. The
reaction was inverse quenched into saturated sodium bicarbonate (10 v) and
extracted with
MTBE (15 v). The extract was washed with brine (5 v), concentrated, then
purified by
chromatography to provide 20 (0.86 wt).
1. dimethylhydroxylamine HCI
TBSO
õõõ
AlMe3, CH2Cl2, 0 C
0
TBSCI, imid. DMF, it 0
OTr
OTr
20 21
[001501 A1Me3 (2 M in toluene, 1.5 eq, 1.5 v) was added to a suspension of
dimethylhydroxylamine hydrogen chloride (1 wt) in CH2C12 (2.5 v) at 0 C. A
solution of
20 (lwt) in CH2C12 (5 v) was added at a rate that maintained the reaction
temperature below
61

CA 02822994 2013-08-01
C. The reaction mixture was then added to aqueous sodium tartrate (L3 M, 20 v)
keeping
the temperature below 10 C. The layers were allowed to partition, were
separated, and the
organic layer was dried with Na2SO4 (5 wt). The resulting suspension was
filtered and the
filtrate concentrated. The residue was dissolved in DMF (2 v) then imidazole
(0.19 wt) and
TBSC1 (0.29 wt) were added. The reaction was diluted with water (5 v) and MTBE
(10 v)
and allowed to stir for 10 minutes. The aqueous layer was removed and the
organic layer
washed with water (5 v). The extract was added to a solution of aqueous NaOH
(IN, 0.78 v)
and Me0H (0.7 v). The reaction was allowed to stir then the aqueous layer was
removed
and the organic layer was washed with brine (2.5 v) then concentrated to
provide 21(1.2
wt).
o
TBp-ON TBISCJ,y
MeMgCI, THF, -20 C
nNOTr
; 21 22
[00151] Methyl magnesium chloride (3.0 M, 59 wt, 1.2 eq) was added to a
solution of 21
(1 wt) in anhydrous THF (1.11 wt, 1.25 v) at a rate that maintained the
reaction temperature
below 0 C. After stirring at 0 C, the reaction was reverse quenched into
saturated
ammonium chloride (2.5 v) and water (2.3 v). The resulting mixture was diluted
with
MTBE (10 v) and stirred vigorously. The aqueous layer was separated and the
organic layer
washed with brine (2.5 v) and concentrated to provide 22 (0.84 wt).
TB,Syty TB,70 OTf."
KHMDS, Tf2NPh, THF
0 -78 to -20 C
OTr
YNNOTr
1 22 23
[00152] Compound 22 (1 wt) was dissolved in THF (4 v) and cooled to -78 C.
KHMDS
(1.5 M in toluene, 1.01 eq, 2.78 wt,) was added while maintaining the
temperature below ¨
60 C. A solution of Tf2NPh (0.62 wt, 1.1 eq) in THF (1.5 v) was added and the
reaction
warmed to -20 C. Saturated ammonium chloride (2.5 v), water (2.5 v), and n-
heptane (2.5
v) were added and the mixture warmed to ambient temperature. The layers were
allowed to
partition and the aqueous layer removed. The organic extract was washed with
saturated
aqueous sodium bicarbonate (3 x 2.5 v) and brine (2.5 v) then concentrated in
vacuo to
provide 23 (1.1 wt).
62

CA 02822994 2013-08-01
-
TBSO OTf OH 0Tf
HCI, IPA, Me0H
0 , 0
n\
OTr OH
23 24
ER-806730
[00153] Compound 23 was dissolved in Me0H (2.5 v) and cooled to 15 C. HC1
(5N in
IPA, 1.30 eq, L18 wt) was added and the resulting solution allowed to warm to
25 C. The
reaction was cooled to 0 C and sodium bicarbonate (3 eq, 0.33 wt) was added.
The reaction
was stirred for 15 minutes and the resulting precipitate removed by
filtration. The filter cake
was washed with ACS grade methanol (1 v) and filtrates were combined and
concentrated.
The crude concentrate was purified by chromatography to provide ER-806730 (24)
(0.5 wt).
Example 4a
[00154] Example 4a provides an alternate method of preparing compounds of
formula A,
an intermediate to F 2, using the general scheme set forth at Scheme V above.
This method
uses ER-812935 as an intermediate as prepared according to Example 3 (compound
4),
above.
0 0
LAH
Me02C,õ0
O 0
THF, 0 C
-0 --CYO
ER-812935 ER-817633
* 9:1 C.23 diastereomers * 9:1 C.23 diastereomers
[00155] ER-812935 (lwt) was dissolved in THF (10 v) and cooled to 0 C. LAH
(1.0 M
in THF, 0.70 eq, 2.0 v) was added keeping the temperature below 5 C. While
stirring
vigorously, excess reagent was quenched with water (0.078 v) keeping the
temperature
below 5 C. While maintaining the vigorous stirring, NaOH (15% wt/wt in water
(0.078 v))
was added followed by water (0.18 v). After adding Celite (2 wt), the
suspension was
filtered and the cake rinsed with THF (5 v). The solution of ER-817633 (0.92
wt, calcd.
based on 100% conversion) was concentrated to 5 v and used directly in the
next stage.
63

CA 02822994 2013-08-01
0 0
MSCI, Et3N
MSOOs" '',0
THF
ER-817633 ER-818937
9:1 C.23 diastereomers *9:1 C.23 diastereomers
[00156] The previously prepared solution of ER-817633 (1 wt in 5 v THF) was
diluted
with THE (5 v), cooled to 5 C and Et3N (3 eq., 0.94 wt) was added. MsC1 (1.05
eq, 0.25 v)
was added at a rate that maintained the temperature below 10 C. The reaction
was
quenched by addition of water (5 wt). Heptane (8 v) was added and the mixture
allowed to
partition. The aqueous phase was separated and extracted with MTBE (2 v). The
combined
organic extracts were washed with saturated sodium bicarbonate (5 v) and water
(1.9 v) The
organic layer was concentrated and solvent exchanged with Et0H to prepare a
solution of
ER-818937 (1.23 wt calcd. based on 100% conversion) in Et0H (1 v) which was
used
directly in the next stage.
KCN
NC->-
Ms0 0"
0.'0--" Et0H, F120 0
0
ER-818937 0-10 70-80 C ER-818950-
' 9:1 C.23 diastereomers 9:1 C.23
diastereomers
[00157] The previously prepared solution of ER-818937 (1 wt in Et0H (0.8 v)
is diluted
with Et0H (190 proof, 9 v). KCN (3 eq., 0.41 wt) was added and the suspension
was heated
to 70-80 C. The reaction was cooled to ambient temperature and water (10 v)
was added
followed by MTBE (10 v). The layers were separated and the aqueous extracted
with
MTBE (5 v). The combined organics are washed with water (2 v) and saturated
brine (4 wt).
The extracts were concentrated and used directly in the next stage.
0
1. AcOH, 1N HCI 0
NC "----==*'-*.>, 0 "0-.40
. 2. recrystalli NC
zation CI 0 '''OH
,
0
OH
ER-818950 ER-817664
9:1 C.23 diastereomers
single diastereomer
[00158] ER-818950 was dissolved in acetic acid (5 v) and hydrogen chloride
(1.0 M, 1
eq, 3 v) was added and the reaction was stirred at ambient temperature. The
reaction was
cooled to 0 C and NaOH (50% wt/wt, 30eq, 7 wt) was added at a rate that
maintained the
temperature below 10 C.
64

CA 02822994 2013-08-01
The solution was extracted with heptane (2 x 10 vol). The aqueous phase was
saturated with
NaCI and extracted with ACN (2 x 10 v). The combined ACN extracts were
concentrated
and solvent exchanged with Et0Ac by atmospheric distillation to provide a
solution of ER-
817664 in Et0Ac (3 v). Salts were filtered from the hot solution which was
then cooled to 0
C. The suspension was filtered to provide ER-817664 as a white crystalline
solid.
Example 4b
[00159] Example 4b provides an alternate method of preparing compounds of
formula F-
2 using the general scheme set forth at Schemes Vb and Vc above. This method
uses ER-
817664 as an intermediate as prepared according to Example 4a, above.
c) \
BrCOC(CH3)20Ac
NCO"s
OH ACN, 0 C 0Ac
-OH Br
ER-817664 C15H20BrN04
Exact Mass: 357.0576
C13H19N04 Mol. Wt.: 358.2276
Exact Mass: 253.1314
Mol. Wt.: 253.2943
[00160] ER-817664 (1 wt) was dissolved in ACN (10 v), the suspension was
cooled to
0 C and 2-acetoxy-2-methylpropanyl bromide (4.0 eq, 2.4 v) was added followed
by the
addition of H20 (1.0 eq., 0.07 v). The resulting mixture was stirred at 0 C
for 2 hours.
NaHCO3 (sat. aqueous, 8.0 eq. 40 v) was added slowly at 0 C. The resulting
mixture was
stirred at room temperature for 30 minutes prior to extraction with MTBE (2 x
20 v). The
organic layer was washed with brine (5 v) and concentrated to give the product
as colorless
oil.
-*X
NC 0 (31'sµ 'OAc Toluen
Q DBU
e NC 0
0 '0Ac
Br
CisFli9N04
C151-120BrN04
Exact Mass: 357_0576 Exact Mass:
277.1314
M
Mol. Wt.: 358.2276 ol. Wt.: 277.3157
[00161] The starting bromide (I wt), depicted immediately above, was
dissolved in
toluene (10 v). DBU (1.8 eq., 0.73 v) was added and the mixture was heated at
80 C. The
mixture was cooled to room temperature, diluted with MTBE (20 v), and washed
with water

CA 02822994 2013-08-01
(5 v) and then brine (5 v). The organic layer was then concentrated to give
the product as
an off-white powder.
/."'==.. 0
1. 03, CH2C12/Me0H
NC 0 NC
..0Ac 2. NaBH, OH
0
ER-818636 ER-818638
3. K2CO3
4. HCI, Na104
[00162] The starting olefin compound (1 wt), depicted immediately above,
was dissolved
in CH2Cl2 (5 v) and Me0H (5 v), and cooled to between ¨40 'V to -45 C. The
solution was
then treated with 03. Excess 03 was removed by N2 purge and the solution was
warmed to ¨
15 C. NaBH4 (1.0 eq, 0.18 wt) was added and the mixture was warmed to 0 C.
K2CO3
(1.3 eq.) was added and the suspension stirred at rt. The reaction was
neutralized with IN
HC1 (-4 eq, ¨20 v) at 0 C and the solution was extracted with MTBE(10 v) to
remove
lypophilics. The aqueous layer was concentrated to remove CH2Cl2 and Me0H. THF
(4 v)
was added followed by NaI04 (2 eq, 2 wt). The reaction was extracted with MTBE
(10 v)
and n-BuOH (10 v). The combined organic extracts were concentrated and the
resulting
powder was triturated with Et0Ac. After filtration the lactol was isolated as
a pale yellow
powder.
(Me0)2POCH2CO2Me
NCO OH
LiCI, i-Pr2NEt NC OH CO2Me
ER-818638 ER-818640
[00163] ER-818638 (lwt) and LiCI (2.0eq, 0.35 wt) was stirred in ACN (8.7
v). Hunig's
base (1.5 eq) was added at 25 C. 1 N HCl (5 v) was added and the mixture was
extracted
with MTBE (10 v). The organics were concentrated to provide ER-818640 which
was used
as is in the next step.
H2, Pd(C)
Me0H
OH OH
C15H21N05 C15H23N05
Exact Mass: 295.142 Exact Mass:
297.1576
Mob. Wt.: 295.3309 Mob. Wt.:
297.3468
[00164] The starting a-olefin ester compound (1 wt), depicted immediately
above, was
dissolved in Me0H ( I 0 v) and added to 10 wt% of Pd(C) (0.09 eq,-0.33 wt)
under N2. The
66

CA 02822994 2013-08-01
suspension was then stirred under H2. The suspension was filtered through a
Celite pad (20
wt), rinsing the filter cake with Me0I-1 (20 v). The filtrate was concentrated
and purified by
flash chromatography to give product as colorless oil (94.3% yield).
NCO
NIS, Ph3P, py
OH 02 Me
C15H23N05 C15H221N04
Exact Mass: 297.1576 Exact Mass: 407.0594
Mol. VVI: 297.3468 Mol. Wt: 407.2439
[00165] Pyridine (10 eq.), Ph3P (7 eq.) and NIS (4 eq.) were added to the
solution of the
ester (1 wt) in THF (15 v) separately. The reaction mixture was stirred at
ambient
temperature. Aqueous citric acid (20 wt%, 10 eq) was added and the mixture
diluted with
TBME (30 v). The aqueous layer was separated and the organic layer washed with
water (5
v), aqueous Na2S203 (10% wt/v, (5 v), water (5 v) and brine (5 v). The organic
layer was
concentrated and purified by flash chromatography to give product as colorless
oil.
NaBH4,
_____________________________________ -
Me0H, 55 C OH
Ci5H221N04
Ci4H221NO3
Exact Mass: 407.0594 Exact Mass: 379.0644
Mol. VVt.: 407.2439 Mol. Wt.: 379.2339
[00166] The starting iodide (1 wt) was dissolved in Me0H (30 v) and heated
to 55 C.
NaBH4 (47 eq.) was added in 6 portions at 55 C over 80 minutes. The reaction
was cooled
to 0 C and quenched with 1N HC1 (30 v). After stirring 5 minutes, the mixture
was diluted
with brine (30 v) and extracted with DCM (50 v x 2). The organic layer was
dried over
Na2SO4 and concentrated. The crude product was used directly in the next step.
0
)21
NCO
Zn, Et0H, 80 C
0
OH OH
Ci4H2204
C1 H221NO3 Exact Mass: 254.1518
Exact Mass: 379.0644 Mol. Wt.: 254.3221
Mol. Wt.: 379.2339
67

CA 02822994 2013-08-01
[001671 The starting alcohol (1 wt), depicted immediately above, was
dissolved in Et0H
(70 v) and Zn (165 eq.) was added. The suspension was refluxed at 75-80 C.
The reaction
mixture was cooled to ambient temperature and IN FICI (70 v) was added. The
mixture was
extracted with DCM (3 x 100 v), the organic layer washed with brine and
concentrated.
TBDPSCI, DMAP, Et3N
0 , 0
_________________ \\OH CH2Cl2
OTBDPS
Ci4H2204
C30H4004St
Exact Mass: 254.1518 Exact Mass: 492_2696
Mol. Wt.: 254.3221 Mol. Wt.: 492.7217
[00168] The starting lactone, as depicted immediately above, was dissolved
in DCM (50
v), Et3N (5.0 eq.), DMAP (0.3 eq.) and TBDPSC1 (1.5 eq.) were added separately
at ambient
temperature under N2, and the resulting solution was stirred at ambient
temperature for 2-3
hours. Upon the completion of the reaction, the mixture was diluted with 1BME
(100 v),
washed with sat. aq. NaHCO3 solution (10 v), H20 (10 v) and brine (10 v). The
organic
layer was concentrated and purified by flash chromatography to give the
product as colorless
oil.
Example 4c
[001691 Example 4c provides another alternate method of preparing compounds
of
formula F-2 using the general scheme set forth at Scheme VII above. This
method uses ER-
811510 as an intermediate as prepared according to Example 3, above where
acetone is used
instead of cyclohexanone.
0
0 0
12)
"0 ______________________________________________ Os'
ER-811510 ER-812771
[001701 ER-811510 (I wt, 1 eq) was dissolved in methylene chloride (6.3 v)
and cooled
to -5 C. Pyridine (0.41 vol, 1.1 eq) was added followed by bromoacetyl bromide
(0.44 vol,
1.1 eq) while keeping the temperature below 0 C. The reaction was stirred at 1
hour and
warmed to room temperature. Water (8 vol) was added and the layers separated.
The
organic layer was washed sequentially with aqueous copper sulfate pentahydrate
(1.0 M, 10
68

CA 02822994 2013-08-01
vol), water (8 vol), and brine (10 vol) then dried over magnesium sulfate,
filtered and
concentrated in vacuo to afford ER-812771 as a tan solid.
0
0 0
"0 '0
ER-812771 ER-812772
[00171] ER-812771 (lwt, 1 eq) was dissolved in acetonitrile (6 v) and
triphenylphosphine was added and the reaction heated at 50 C for 45 minutes.
The reaction
was cooled to -10 C then 1,8-diazabicyclo[5.4.0jundec-7-ene (DBU, 0.35 vol,
0.8 eq) was
added. The reaction was stirred for 15 minutes, heated to 80 C for 45 minutes
then cooled to
ambient temperature. Ammonium chloride (saturated aqueous, 10 vol) was added
and the
aqueous layer extracted with ethyl acetate (3 x 10 v). The combined organic
layers were
dried over magnesium sulfate and concentrated in vacuo. The crude product was
purified by
chromatography to afford ER-812772 as a white solid.
0 " 0
0
ER-812772 ER-812829
[00172] ER-812772 (1 wt, 1 eq) was dissolved in ethyl acetate (8 v). 10%
Palladium on
carbon (0.05 wt, 0.01 eq) was added, the reaction purged with nitrogen then
stirred under
hydrogen atmosphere for 2 hours. The catalyst was removed by filtration
through Celite
with ethyl acetate washed. The combined filtrates were concentrated in vacuo
to afford ER-
812829 as a white solid.
69

CA 02822994 2013-08-01
Example 5
Preparation of F-3a:
H
H OH
OzOtOH

cyclohexanone 5(0,
HO OH p-TSA, toluene
ER-805813 ER-805715
[00173] D-Gulonolactone (1 wt., 1 eq.), cyclohexanone (2 to 3 eq.), toluene
(6 vol.), and
p-toluenesulfonic acid (0.021 wt., 0.02 eq.) were charged to the reaction
vessel. Reaction
mixture was heated to reflux with stirring. Upon azeotropic removal of water
the reaction
was complete. The reaction mixture was cooled to 85 to 90 C and agitation was
increased.
Heptane (5.2 vol.) was added over 20-30 minutes with stirring. Cooled to 65-70
C and
stirred for 30 minutes at 65-70 C. The solid product was filtered at 65-70 C,
maintaining
mother liquor temp > 35 C. Re-filtered at 35-40 C and maintained the mother
liquor at
ambient temperature for 30 minutes. Re-filtered the mother liquor. The filter
cake was
washed two times with heptane (2 x 1.7 vol.) then dried to afford ER-8057I5.
Yield 84%
(1.6 wt.).
[00174] In an alternate method for preparing ER-805715, D-gulonolactone (1
wt),
cyclohexanone (1.32 wt, 2.4 eq), p-Ts0H-monohydrate (0.02 wt, 0.02 eq) and
toluene (12
vol) were refluxed together for 19 hours, while azeotropically removing water.
The mixture
was washed with 5% aqueous NaHCO3 (4 vol) followed by saturated aqueous NaCl
(2 vol x
2, pH=7). The organic phase was concentrated by distillation (ca. 4.5 vol of
toluene
remaining) and cooled to 100 C before heptane (10 vol) was added, maintaining
internal
temperature > 80 C. The mixture was heated to reflux for at least 1 hour
before it was
cooled to and aged at 85 C for 3 hours, at 80 C for 3 hrs and then cooled to
40 C in 12 hrs.
The product was collected by filtration and the cake washed with heptane (2
vol). The filter
cake was dried by airflow to afford ER-805715 (1.48 wt) in 78% of yield.

CA 02822994 2013-08-01
H OQ H
0
DIBALEI 5<-)
toiuenefITIF
ER-805715 ER-805814
[00175] ER-805715 (1 wt., 1 eq.) was charged to reaction vessel and
dissolved in
anhydrous THF (3.34 vol.) and anhydrous toluene (2.5 vol.). The mixture was
cooled to ¨15
to -10 C. DIBALH (1.5M in toluene, 2.4 vol., 1.2 eq.) was added over 1 hour
and the
mixture stirred for 15-30 minutes at ¨15 to ¨10 C. The reaction was inverse
quenched into
KNa-Tartrate solution (1 wt. KNa Tartrate in 2.9 wt. water) at 10 C and the
resulting =
mixture allowed to warm to room temperature and stir for 4 hours. The mixture
was filtered
then the layers separated and extracted with MTBE (2 vol.). The organic layers
were
combined and the solvents removed in vacuo to afford ER-805814. Yield 100%,
(1.02 Wt.).
H PP Me0 HO p---0
HO 0
Orx,0 KOtBu, THF,
Ph3P CH20Me
ER-805814 ER-805815
[00176] ER 805814 (1 wt.) was dissolved in anhydrous THF (3.3 vol.) and
treated with
(methoxymethyl)triphenylphosphonium chloride (2.11 wt., 2.1 eq.). The reaction
mixture
was heated to 28-32 C then a solution of KOtBu (0.66 wt., 2 eq.) in anhydrous
THF (2.64
vol.) was added over 100-140 minutes, maintaining reaction temperature 30-35
C. After 5
hours, the reaction was cooled to 20-25 C, MTBE (5.11 vol.) was added and the
mixture
stirred. Brine (3 wt.) and water (3 wt.) were added (exothermic at start of
addition,
controlled by bath @ 20-25 C). Organic layer was separated and treated with a
solution of
maleic anhydride (0.27 wt.) in MTBE/THF (1/1 v/v, 1.78 vol.). NaOH solution
(0.088 wt. in
2.5 vol. water) was added slowly to the reaction mixture. The organic layer
was
concentrated to give crude ER 805815 (0.985 wt.). The residue was triturated
three times
with MTBE/heptane (1/4 v/v, 6.6 vol.). The extract was filtered through S102
(3 wt), eluting
with MTBE/heptane (1/2 v/v, 45 vol.). The filtrate was concentrated to give ER-
805815
(0.88 wt., 81% yield).
71

CA 02822994 2013-08-01
[00177] In an alternate method for preparing ER-8058I5, a solution of t-
BuOK (0.989
wt, 3 eq) in THF (4 wt) was added to a suspension of
(methoxymethyl)triphenylphosphonium chloride (3.12 wt, 3.1 eq) in THF (1.78
wt),
maintaining the reaction temperature between 0-10 C. The addition vessel was
rinsed with
THF (2 x 0.7 wt). A solution of ER-805814 (1 wt, 1 eq) in THF (1.42 wt) was
added to the
reaction, maintaining 0-10 'C. The addition vessel was rinsed with THY' (2 x
0.7 wt). The
mixture was stirred at 20-30 C overnight and 30-35 C for 3 hours. The
reaction was
cooled below 30 C and diluted with MTBE (3.7 wt) followed by 10 wt% aqueous
NaC1(4
wt) solution. The mixture was stirred for 30 minutes and the layers were
separated. Maleic
anhydride (0.63 wt, 2.2 eq) was added and the mixture stirred at room
temperature for 30
minutes. Water (6 wt) and a solution of NaOH (48 wt%, 0.64 wt, 2.6 eq) was
added
dropwise, maintaining the reaction below 15 C. After stirring below 15 C,
the lower layer
was separated. Water (6 wt) was added followed by a solution of NaOH (48 wt%,
0.64 wt,
2.6 eq), keeping the mixture below 15 C during the addition. After stirring
below 15 C,
the lower layer was separated. The organic layer was washed three times with a
15 wt%
aqueous NaCI solution (3 x 4 wt). The organic layer was concentrated in vacuo.
The
residue was diluted with MTBE (1 wt) and concentrated in vacuo. The residue
was diluted
dropwise with IPE (3 wt) at 40-50 C over 30 minutes. The suspension was
stirred for 1
hour at 40-50 C and slowly cooled to 0-10 C and stirred for 1 hour. The
solids were
filtered and the cake washed with IPE (2 wt). The filtrate and washings were
concentrated
in vacuo. The residue was treated with Me0H (2.37 wt) and water (0.4 wt) and
extracted
with heptane (2.74 wt). The lower layer was extracted 9 times with heptane
(2.05 wt). The
extracted solutions were combined and concentrated in vacuo to give ER-805815
(1.07 wt,
98.6%).
[00178] In an alternative method for workup of ER-805815, the crude organic
layer that
is produced following brine wash and concentration is treated with MTBE (2.86
wt) and
celite (0.5 wt). After stirring for 2.5 h, heptane (1.46 wt) was added over 2
hrs and the
mixture stirred overnight. The precipitate was filtered. The filter cake was
washed with
MTBE/Heptane (1:1) (5 wt). The filtrate was concentrated in vacuo until the
volume was
decreased to about 3 volume. The residue was dissolved in Me0H (2 wts) and H20
(6 wts).
The mixture was extracted with heptane/MTBE (5:1) (3*6 wts). The organic layer
was
separated and concentrated to provide ER-805815 which was used as is for the
following
step.
72

CA 02822994 2013-08-01
Me0 HO co-0 H 9C1)

0s04, NMO, acetone-
water
On
ER-805815 ER-805816
[00179] ER-805815 (1 wt) was dissolved in acetone (2.4 vol) and water (0.4
vol).
N-Methylmorpholine N-oxide (0.62 wt, 2 eq) was added and the mixture cooled to
0-5 'C.
0s04 (0.15M in water, 0.065 vol) was added and the reaction was maintained at
0-5 C.
The reaction mixture was stirred at 0-5 C for 12 hours. Water (0.2 vol) was
added over 1
hour at 0-2 C. The mixture was stirred for one hour at 0-5 C. The product was
filtered and
the solids washed twice with pre-cooled (0-5 C) acetone/water (1/1, v/v, 2 x
0.7 vol). The
product was dried to afford ER-805816 (0.526 wt, 52% yield, residual Os <17
ppm).
[00180] In an alternate method for preparing ER-805816, a solution of ER-
805815 (1 wt,
1 eq) in acetone (4 wt) was charged into a four-necked flask, then water (0.5
wt) was added
at ambient temperature. To the mixture was added anhydrous N-methylmorpholine-
N-oxide
(0.38 wt, 1.2 eq). Potassium osmate dihydrate (0.003 wt, 0.003 eq) was added
portion-wise
at 25 to 35 'V while cooling with water. The mixture was kept at this
temperature for 4
hours. A solution of sodium thiosulfate 0.075 wt, 0.49 eq) in water (0.5 wt)
was added at
ambient temperature, then the mixture was stirred for 0.5 hour. The mixture
was cooled to 0-
C and stirred for 2 hours. The resulting precipitate was collected and the wet
cake was
washed with methanol (0.6 wt) and water (1.5 wt) to obtain the crude product
(1.25 wt). The
crude product sample was dried (0.611 wt). The crude ER-805816 (1.25 wt) was
added to
water (3.05 wt) and stirred for 2 hours at about 25 C. The precipitate was
filtered and
washed with water (1.53 wt) to afford the crude wet cake (1.05 wt). The crude
product
sample was dried and sampled, ICP Os = 37 ppm. The crude ER-805816 (1.05 wt)
was
added to water (2.81 wt) and stirred for 2 hours at about 25 C. The
precipitate was filtered
and washed with water (1.4 wt) and methanol (0A5 wt) to afford the crude ER-
805816
(0.736 wt). The wet cake was dried crude product (0.56 wt, ICP (Os) = 28 ppm).
ER.-
805816 (0.56 wt) was dissolved in acetone (1.76 wt) at 45 to 55 C. To the
solution was
added active carbon (0.027 wt) and stirred at same temperature for 0.5 hour.
The mixture
was filtered and the cake was
73

CA 02822994 2013-08-01
washed with hot acetone (0.214 wt). The filtrate was kept at 45 to 50 C and
water (0.83 wt)
was added over 10 minutes and temperature was kept at 40 to 50 C during water
addition.
The mixture was cooled to 0 to 5 C and stirred for 1.5 hours. The white
precipitate was
filtered and washed with a solution of acetone (0.17 wt) and water (0.22 wt)
then dried to
give ER-805816 (0.508 wt, 0.49 eq, KF 5.0%, ICP (Os) 9.6 ppm).
H-9-C) H Ac
E OAc
HOo Ac20, AcOH, ZnCl2 AGO
ER-805816 ER-805819
[00181] ER-805816 (1 wt) was slurried in acetic acid (0.89 vol, 5.8 eq) and
acetic
anhydride (3.57 wt, 13 eq). Anhydrous ZnCl2 (0.2 wt, 0.54 eq) was added.
Reaction mixture
was stirred for 24 hours 18-22 C. Reaction was quenched into ice (5 wt) and
water (5 vol).
Et0Ac (10 vol) was added with stirring and the aqueous layer is separated. The
aqueous
layer was back extracted with Et0Ac (10 vol). The combined organic layers were
washed
sequentially with brine (10 vol), 5% aqueous Na0Ac (6 vol), and brine (6 vol).
The organic
layer was concentrated. The crude concentrate was dissolved in 25% Et0Ac/hex
(4 vol) and
filtered through Si02. The pad was washed with 25% Et0Ac/hex (2 x 12 vol) and
further
25% Et0Ac/hex (48 vol). The organic layer was concentrated to give ER-805819
(1 wt,
81%).
[00182] In an alternate method for preparing ER-805819, zinc chloride (0.2
wt, 0.54 eq),
acetic anhydride (2.75 wt, 10 eq), and acetic acid (1 wt, 6 eq) were combined.
The mixture
was cooled to 15-20 C. ER-805816 (1 wt, 1 eq) was added, maintaining the
internal
temperature at 15 to 30 C. The mixture was then stirred at 35-40 C for 6
hours. The
reaction mixture was cooled below 25 'C. Methanol (3.2 wt, 4 vol) was added
drop-wise
maintaining reaction temperature below 25 C. Heptane (2.7 wt, 4 vol) was
added. Water
was added (4 wt, 4 vol) maintaining reaction temperature below 25 C. The
mixture was
stirred for 15 minutes, and then the phases were separated. The lower layer
was washed
twice with heptane (2.7 wt, 4 vol) and the heptane layers were discarded. The
lower layer
was extracted twice with toluene (6.1 wt, 8 vol). The combined toluene layers
were washed
74

CA 02822994 2013-08-01
twice with 17 wt % potassium bicarbonate aqueous solution (0.82 wt KHCO3 in
3.98 wt
water, 4.36 vol), twice with water (4 wt), and concentrated. Methanol (3.95
wt, 5 vol) was
added at 25-30 C and the mixture stirred for 10 minutes. Water (0.3 wt) was
added at 25-30
C. The mixture was cooled to 0 C and seeded. The mixture was stirred at 0 C
for 1 hour.
Water (0.7 wt) was added drop-wise over 1 hour. Water (4 wt) was added drop-
wise over 1
hour. The resulting precipitated solids were filtered, and the filter cake
washed twice with a 0
C methanol (1.03 wt) and water (0.7 wt) solution. The cake was dried to afford
ER-805819
(0.99wt, 0.84 eq).
H
OAc OAc
AcO O: OAc

Me02C,-,õµ,,C1 OAc
AcO"C-Cµ0 Me0 C
2 Ac0 0
TMS on
BF3.0Et, CH3CN
ER-805819 ER-805821
[00183] ER-805819 (1 wt) was dissolved in anhydrous acetonitrile (15 vol)
and treated
with methyl 3-trimethylsilylpent-4-eneoate (0.93 vol, 2 eq). The reaction
mixture was
cooled to 0 -5 C and BF3.0Et2 (0.54 vol, 1.95 eq) was added over 5 minutes,
maintaining
reaction temperature between 0-5 C. Reaction mixture was stirred 0-5 C for
12 hours.
Reaction was quenched into saturated sodium bicarbonate (20 vol) with vigorous
stirring.
Extracted twice with Et0Ac (2 x 8 vol). The combined organics were washed with
brine (12
vol) and concentrated to give ER-805821 (1 wt, 88% yield, use as is).
[00184] In an alternate method for preparing ER-805821, ER 805819 (1 wt, 1
eq) and
methyl 3-trimethylsilylpent-4-eneoate (0.93 vol, 2 eq) were dissolved in
anhydrous
acetonitrile (5.46 wt, 7 vol). The reaction mixture was cooled to 0-5 C and
BF3-0Et2 (0.54
vol, 1.95 eq) was added over 5 minutes, while maintaining reaction temperature
between 0-5
'C. The reaction mixture was stirred at 0-5 C for 20 hours then heptane (5.47
wt, 8 vol)
was added at 0-5 C. The phases were separated and the lower layer treated
with heptane
(5.47 wt, 8 vol) at 0-5 C. The reaction was quenched by dropwise addition of
7.4%
potassium bicarbonate aqueous solution (0.64 wt KHCO3 and 8 wt water), while
maintaining
the reaction temperature at 0-15 C. Toluene (8.65 wt, 10 vol) was added and
the mixture
stirred for 30 minutes. The lower layer was separated and the upper organic
layer washed
twice with water (10 vol) and concentrated to afford ER-805821 as a crude oil
(1.05 wt,
0.935 eq).

CA 02822994 2013-08-01
OAc OH
OAc
Base,
Ac00 Solvent Me02C,_,o
ob H
ER-805821 ER-805822
[00185] ER-805821 (1 wt) was dissolved in anhydrous THF (8.4 vol) and
anhydrous
Me0Ac (2 vol). Triton B(OH) (3.6 vol) was added over 2 minutes, reaction
maintained 17-
23 C. Reaction was stirred for 1.5 hour. Reaction mixture was filtered. The
filtrate was
concentrated and passed through a pad of Si02 (5 wt, Et0Ae, 20 vol). The
filtrate was
washed with brine (2.2 vol) and evaporated to give ER-805822 (0.54 wt, 72%
yield).
[00186] In an alternate method for preparing ER-805822, ER-805821 (1 wt, 1
eq, 11.18
g, 21.81 mmol) was dissolved in anhydrous MTBE (4.4 wt, 6 vol.) and cooled to
0-5 C.
Na0Me (28wt% in Me0H, 0.564 wt, 1.5 eq) was added to the mixture over 1 hour
at 0-5 C
and stirred for 3 hour at same temperature range. The reaction was quenched by
addition of
acetic acid (0.188 wt, 1.6 eq.), maintaining 0-5 C during addition. The
mixture was stirred
overnight then treated with a 5 wt % aqueous solution of KHCO3 (3 wt) and
ethyl acetate
(3.6 wt, 4 vol) at 0 -5 C, and then stirred for 15 minutes. After phase
separation, the lower
layer was extracted with ethyl acetate (3.6 wt, 4 vol) twice. The combined
organic layer was
concentrated. To the residue was added acetone (2 wt, 2.5 vol) and IPE (2 wt,
2.7 vol) and
stirred overnight at 0-5 C. The mixture was filtered through Celite (0.25
wt) and washed
with acetone (2 wt). The filtrate was concentrated to afford the crude oil
(0.55 wt). To the
residue was added acetone (0.2 wt, 0.25 vol.) and IPE (0.54 wt, 0.75 vol.) and
stirred for 1
hour at 40-50 C. The solution was seeded with ER-805822 at room temperature
and stirred
overnight at room temperature. To the suspension was added IPE (1.27 wt, 1.75
vol.) over 2
hours at room temperature. After stirring for 5 hours at room temperature, the
precipitate
was collected by filtration, and the cake washed with acetone/IPE (1/10) (2
vol.). The
obtained cake was dried in a tray-type chamber at 30-40 C overnight to afford
the desired
product ER-805822 (0.286 wt, 0.38 eq) in 38.0 % yield from ER-805819.
[00187] In an alternative method for workup of crude ER-805822, the residue
following
concentration of the final Et0Ac solution was dissolved in IPA (2 wt) and the
solution was
heated to 50 C. Heptane (5 wts) was added and the mixture was cooled to 20 C
and
seeded.
76

CA 02822994 2013-08-01
The mixture was stirred at 20 C overnight. Heptane (10 wts) was added and the
mixture was
cooled to -5 C in 30 min and stirred at -5 C for 5 hrs. The mixture was
filtered and the filter
cake was washed with heptane (2 wts). The filter cake was dried with air flow
under vacuum
to provide ER-805822 (60%).
OOH
OH 0
C
02 0
0 Na104, solvent, Me 0
Oo water
Methyl Ester Diol Aldehyde
ER-805822 ER-804697
[00188] ER 805822 (1 wt) was dissolved in ethyl acetate or another
appropriate solvent
(5 vol) and water (5 vol). NaI04 (0.58 wt, 1.05 eq) is added portionwise over
30 min to 1
hour, maintaining reaction temperature 0-10 C. Reaction is stirred for up to
2 hours. The
reaction mixture was treated with NaC1 (1 wt) and stirred for 30 min at 0 to
10 C. The
reaction mixture was filtered and the cake is rinsed with with ethyl acetate
(2 vol). The
phases were separated and the lower layer extracted with Et0Ac (5 vol) three
times. The
combined organic layer was washed with 20% aqueous NaC1 (5 wt). The organic
layer was
concentrated to give ER 804697 (1 wt). The residue was dissolved in toluene (2
vol) and
the solution concentrated. The residue was dissolved in acetonitrile (7 vol)
and used for the
next step.
0 Ei OH
H H
C)CTMS
IVIe0 2 0 2
Me0C
NiCl2, CrCl2
0-t-) Br.0O
TMS
Aldehyde Vinyl Silane
ER-804697 ER-804698
[001891 NiCl2 (0.025 wt) and CrC12 (2.5 wt) were charged to reaction vessel
under inert
atmosphere. Anhydrous dichloromethane (5 vol) was charged. Stirring was
initiated and the
mixture was cooled to 0-3 'C. Anhydrous DMSO (6.7 vol) was added with vigorous
stirring
over 45 minutes, maintaining temperature below 20 C. ER-804697 (1 wt) was
dissolved in
anhydrous dichloromethane (1 vol) and charged to the reaction vessel. The
resulting
mixture
77

CA 02822994 2013-08-01
was warmed to 25 C and 1-bromo-2-trimethylsilylethylene (2.58 wt) was added
neat over
20 minutes. The reaction temperature was maintained below 45 C. The reaction
was stirred
for 30 minutes at 25-35 C following complete addition. Methanol (5 vol) was
added and the
mixture was stirred for 10 minutes. MTBE (33 vol) was charged and the slurry
transferred
into IN HCI (25 vol) and water (10 vol). The mixture was stirred for 5
minutes. The
aqueous layer was back extracted with MTBE (10 vol) and the combined organics
washed
sequentially with 0.2N HCI (17 vol), twice with 1% NaCI solution (2 x 17 vol),
and brine (13
vol). The organic layer was concentrated and purified (Si02, 25 wt, 10 column
vol
Et0Ac/Hex 1/3.5 v/v) to give ER-804698 (0.53 wt, 61%).
[00190] In an alternate method, this reaction was performed in the presence
of the chiral
ligand ER-807363 in a manner substantially similar to that described for the
preparation of
ER-118047, infra.
[00191] In an alternate method for preparing ER-804698, DMSO (7 vol.) and
MeCN (7
vol) were degassed and cooled to 0-10 C. The solution was treated portionwise
with CrC12
(10 eq, 3.47 wt) and NiC12 (0.1 eq, 0.037 wt) such that the internal
temperature did not
exceed 20 C. A solution of ER-804697 (1 wt, 1 eq) in MeCN (7 vol) and 1-bromo-
2-
trimethylsilylethylene (5 eq, 2.5 wt) were added dropwise at 0-10 C, not
allowing the
internal temperature to exceed 15 'C. The reaction mixture was stirred at 5 -
15 C overnight.
To the mixture was added methanol (5.5 wt), water (7 wts), and MTBE (5.2 wts).
The
reaction was stirred for I hour and the lower layer was separated (layer 1).
To the upper
layer was added a premixed solution of NaC1 (1.5 wts) and water (13.5 wts).
The mixture
was stirred for 1 hour and the lower layer was separated (layer 2). To the
upper layer was
added heptane (4.8 wts), methanol (2.8 wts), and a premixed solution of NaC1
(1.5 wts) and
water (13.5 wts). The mixture was stirred for I hour and the lower layer was
separated
(layer 3). The upper layer was drained and saved (organic 1). The reactor was
charged with
layer 1, methanol (2.8 wts), and MTBE (2.8 wts). The mixture was stirred
overnight. The
lower layer was separated and discarded. The upper layer was treated with
layer 2. The
mixture was stirred for 1 hour and the lower layer was separated and
discarded. The upper
layer was treated with layer 3 and heptane (4.8 wts). The mixture was stirred
for 1 hour and
the lower layer was separated and discarded. The upper layer was drained and
saved
(organic 2). The reactor was charged with layer 3, MTBE (0.8 wts), and heptanc
(2.7 wts).
The mixture was stirred for 1 hour and the lower layers was separated and
discarded. The
upper layer was combined with organic 1 and organic 2. The combined organics
were
78

CA 02822994 2013-08-01
filtered and concentrated at reduced pressure to afford the crude ER-804698
which was
purified by chromatography (Si02, 25 wt, 10 column vol Et0Ac/Hex 1/3.5 v/v )
to give ER-
804698 (0.67 wt, 57% yield).
[00192] In an alternate method of preparation of ER-804698, the crude
material is taken
directly to the next step without purification.
OH OH
H H H 0Hi
TMS TMS
Me02C
AcOH, water
OO OH
ER-804698 ER-807023
[00193] ER 804698 (1 wt, 1 eq) was treated with AcOH (4.2 wts) and water
(4.2 wts).
The mixture was heated to 90-97 C for 100 min. The mixture was cooled to
below 15 C
then washed with heptane (2 x 2.7 wts) twice below 15 C. After phase
separation, a mixture
of 20wt% aqueous KFIC03 solution (7.7 wts, 35 eq) and MTBE (5.95 wts) was
added
dropwise to the lower layer such that temperature does not exceed 15 C. After
phase
separation the upper layer was washed successively with 5wt% aqueous KHCO3
solution
(0.2 wts), and twice with 5wt% aqueous NaCI solution (2 x 0.2 wts). The
organic layer was
concentrated under reduced pressure and MTBE (1.49 wts) was added. The mixture
was
heated to 55 C and stirred until dissolved. Heptane (1.00 wts) was added to
the solution and
the solution was cooled to 40-45 C. Additional heptane (4.47 wts) was added
to the
solution and the solution was cooled to 5-15 C and then stirred overnight.
The crystals
were filtered and rinsed with heptane to provide ER-807023 (0.58 wts, 71%
yield).
OH OTBS
H H
H 0Hi
TMS TBSOTf, 2,6-lutidine, TMS
0
MTBE Me02C - OH
111
OHOTBS
OTBS
ER-807023 ER-804699
[001941 ER-807023 (1 wt, 1 eq) and MTBE (7.43 wts) were charged to a
reactor under a
nitrogen atmosphere. To the reaction was added 2,6-lutidine (2.15 wts, 7.5
eq). To the
mixture was added dropwise TBSOTf (2.47 wts, 3.5 eq) at 0 C. The reaction
mixture was
stirred for 30 min at 0-10 C, then warmed to 23 C over 1 hr and held at 23
C for 16 hrs.
79

CA 02822994 2013-08-01
Me0H (0.21 wts, 2.5 eq.) and water (14.8 wts) were added dropwise sequentially
to the
reaction mixture, maintaining temperature below 30 C. After phase separation,
the upper
layer was washed with IN aqueous hydrochloric acid (16.2 ms), 5% NaCI aq.
(14.8 wts),
5% NaHCO3 aq. (14.8 wts), 5% NaCI aq. (14.8 wts), and 5% NaCI aq. (14.8 wts),
respectively. The upper organic layer was concentrated by distillation under
reduced
pressure to afford the crude ER-804699. Me0H (7.91 wts) was added and the
mixture was
heated to 50 C for 30 min. The mixture was cooled to 0 C over 5 h, and then
stirred
overnight at 0 C. The solid was filtered, and the cake was washed with cold
Me0H (4 wts)
and dried to yield ER 804699 (1.42 wts, 74% yield).
OTBS OTBS
H H 0
TMS ______________________________________________________________ I
NIS, toluene, ACN Me02C
I:1 OTBS 0 ----1(µOTBS
OTBS OTBS
ER-804699 ER-803895
[00195] Into a reactor under a nitrogen atmosphere was charged a solution
of ER-804699
(1 wt, 1 eq) in toluene (2.60 wts). Acetonitrile (4.72 wts) was added. TBSC1
(0.011 wts,
0.05 eq) was added. The reaction mixture was warmed to 30 C and the NIS was
added
(1.25 wts, 4 eq). The reaction mixture was stirred at 22 hrs at 30 C. The
reaction was cooled
to 25 C and the mixture of aqueous sodium thiosulfate and sodium bicarbonate
(10.35 wts)
were added over 10 minutes keeping the internal temperature below 30 C. The
reaction
was stirred for 30 minutes at 25 C. The aqueous layer was separated. The
upper layer was
washed twice with 10% NaCI (aq) (2 x 9.9 wts). The organic layer was
concentrated under
reduced pressure to give crude ER-803895 that was purified using silica gel
chromatography
to provide ER-803895 (0.96 wt, 89.5% yield).

CA 02822994 2013-08-01
Example 6
Assembly of F-la, F-2a, and F-3a and Preparation of B-1939:
A. Preparation of (R) or (S) N-12-(4-Isopropyl-4,5-dihydro-oxazol-2-y1)-6-
methyl-
phenyl] -methanesulfonamide:
0
COOH (CI3C0)2C0
INNH
2 ?
THF 00 to 25 C, 18h N4C)
Me
Me
ER-807244 ER-807245
[00196] A pre-dried glass lined reactor was charged with triphosgene (1
wt., 1 eq.) and
anhydrous THF (2 vol.) and was cooled to an internal temperature of -10 C. A
second pre-
dried glass lined was charged with ER-807244 (1.27 wt., 2.5 eq.) and anhydrous
THE (3
vol.) then cooled to an internal temperature of -10 C. The contents of the
first reactor were
transferred into the second reactor at a rate such that internal temperature
did not exceed 15
C. After complete addition, the reaction was stirred at an internal
temperature of 0 C for 1
hour and then gradually warmed to 25 C. A sparge of nitrogen was used for 18
hours to
scrub away excess phosgene with trapping of the off-gases through a 2 N NaOH
solution.
MTBE (3 vol.) was added and the solvent removed by distillation under N2 purge
at 40 to
46 C, adding more MTBE as heeded. Upon complete removal of the phosgene, the
mixture
was cooled to an internal temperature of 5 to 10 C and the solution filtered
with MTBE (3
vol.) washes to yield ER-807245 (1.12 wt., 0.97 eq.) as a white crystalline
solid.
0
1) DMF, 90 C, 2h
?
Li el Me
_____________________________________________ )0--
NO
HO,- 2) Li0H.H20, 60 C, 2h HO 0 NH2
Me 65% to 75%
ER-807245 D-valinol or L-valinol ER-806628 (D-valinol)
ER-808056 (L-valinol)
[00197] Into a pre-dried and inerted reactor 1, was added ER-807245 (1 wt.,
1 eq.) and
anhydrous DMF (4 vol.). With stirring, the mixture was heated to an internal
temperature of
95 C. D or L-Valinol (1.05 eq., 0.61 wt.) was dissolved in anhydrous (DMF 1.3
vol.) in
reactor 2 with heating to an internal temperature of 90 C. The contents of
reactor 2 were
transferred into reactor 1 at internal temperature 90 C. CO2 evolution was be
observed and
the reaction was vented with a N2 bleed. The reaction solution was stirred at
90 C for 3
81

CA 02822994 2013-08-01
hours and then cooled to an internal temperature of 65 C. Then, an aqueous
slurry of
lithium hydroxide (0.47 wt., 2 eq.) in water (2 vol.) was added to reactor 1
and the
suspension stirred at an internal temperature of 65 C for 1 hour. The reactor
was charged
with water (5 vol.) cooled to an internal temperature of ¨5 C over 3 hours.
The mixture
was stirred for 8 hours at internal temperature ¨5 C and the desired product
collected by
filtration with water (2 X 4 vol.) washes followed by n-heptane (2 X 3 vol.).
The product
was dried under vacuum and N2 flow at 35 C for 24 hours or until KF 250 ppm
to yield
ER-806628 or ER-808056 (0.80 wt., 0.60 eq.) as a crystalline solid.
MsCI, Pyr, DMAP
NN
110
_________________________________________ VP-
Me 0 C to 25 C me
HO -7- 0 NH2 0 NHMs
ER-806628 (R) ER-806629 (R)
ER-808056 (S) ER-807363 (S)
[00198] A pre-dried and inerted reactor under nitrogen was charged with ER-
806628 or
ER-808056 (1 wt., 1 eq.), pyridine (3 wt., 11.4 eq.) and DMAP (0_03 wt., 0.05
eq.). The
reaction was cooled to an internal temperature of-10 C then methanesulfonyl
chloride (1.46
wt., 3 eq.) was added at a rate such that internal temperature was below 15 C.
Upon
complete addition, the reaction was stirred at an internal temperature of 0-15
C for 1 hour
and then slowly warmed to 25 C over 2 hours. MTBE (2.6 vol.), was added
followed by
process water (2 vol.) at a rate such that the internal temperature did not
exceed 35 C. The
biphasic mixture was titrated with 6N hydrochloric acid, (-1.9 vol.) portion-
wise until the
pH of the aqueous layer= ¨3 to 5_ If pH went under 3, 30% (w/w) aqueous
solution of
Na2CO3 was added to back titrate to the desired pH. The phases were allowed to
partition
and the aqueous phase separated. All organics were combined with water (0.7
vol.) and the
aqueous phase discarded. The MTBE was distilled to a level of-2 vol. at
atmosphere
pressure to constant bp 55 C and KF < 500 ppm. Additional MFBE was added if
necessary.
The solution was cooled to an internal temperature of 5-10 C with seeding
when necessary
to induce crystallization. n-Heptane (0.5 vol.) was added and the mixture
stirred for 18
hours at 5 C. ER-806629 or ER-807363 was collected by filtration with n-
heptane (2 X 3
vol.) washes. A second crop of crystals was obtained by concentration of the
filtrates to V2
volume and cooling to 0 C. The filter cake was dried under N2 for 18 hours.
The crude
82

CA 02822994 2013-08-01
weighted ER-806629 was charged into a pre-dried reactor and MTBE (3 vol.) was
added.
The resulting mixture was heated to an internal temperature of 45-50 C for 45
minutes and
then slowly cooled to 5 C over 3 hours, with seeding when necessary. n-Heptane
(0.5 vol.)
was added and the mixture stirred for 18 hours at an internal temperature of 5
C. The solid
product was collected via filtration and n-heptane (2 X 3 vol.) washes then
dried under
vacuum at 35 C for 24 hours to yield ER-806629 or ER-807363 (1.7 wt., 0.57
eq.) as a
crystalline solid.
B. Assembly of F- I a and F-2a and Intrarnolecular Ether Formation:
TBSO
TBSO __________________________________________________ S02Ph
TBSO,..,K,õsc TBSOõssc
0 0 0
CrC12, NiC12
ER-806067
HO
THE Et3 N
Ms0
Ms0
= \
N

NHMs 0
0
""; NTOPv
___________ ER-805973
ER-807363 ER-808227
[00199] An appropriately sized reactor 1 was charged with ER-807363 (1.82
wt, 3.55 eq)
and the atmosphere was exchanged for nitrogen. Anhydrous THE (15 vol) was
added. In
reactor 2, ER-806067 (F-la, 1.14 wt, 1.1 eq) and ER-805973 (F-2a, 1 wt, 1 eq)
were
combined and dissolved in anhydrous THF (6.3 vol). With stirring, both
reactors were
sparged with nitrogen for 30-45 minutes. Under an inert atmosphere, reactor 2
was charged
with CrC12 (0.75 wt, 3.55 eq) and then heated to an internal temperature of 30
C. Reactor 2
was charged with triethylamine (0.62 wt, 3.55 eq) at a rate such that internal
temperature did
not exceed 45 C. After complete addition, an internal temperature of 30 C
was maintained
for 1 hour. After 1 hour, reactor 2 was cooled to 0 C and charged in an inert
fashion with
NiC12 (0.02 wt, 0.1 eq), followed by the contents of reactor 1 and the
reaction was warmed
to rt. Reactor 2 was cooled to an internal temperature of 0 C and then
ethylenediamine (1.2
vol, 10 eq) was added at a rate such that the internal temperature did not
exceed 10 C.
Note: An exotherm was observed. The reaction was stirred for 1 hour, and then
water (8
vol) and n-heptane (20 vol) were added and the biphasic mixture stirred for 4
minutes and
the layers allowed to partition. The organic layer was separated and the
aqueous layer back
83

CA 02822994 2013-08-01
extracted with MTBE (20 vol). The combined organic layers were concentrated in
vacuo to
a crude oil followed by an azeotrope with anhydrous THF (2 x 10.5 vol). The
crude product
was dissolved in anhydrous THF (4.5 vol) and then stored at ¨20 C until
utilization in the
next stage.
so2ph
TBSO (----SO2Ph
TBSO TBSO __
TBSOõ,_,J..õ. soTBSOc soc
1-SO2Ph
= 0 = 0

=,
KHMDS Chromatography
Ms0 0
THF, -14*C õo
.=5 "5
ER-808227 ER-806746 ER-
804027
100200] The ER-808227 / THF solution from the previous step was analyzed via
KF
analysis. If K.F< 1000 ppm, then proceeded. If KF > 1000 ppm, azeotroped in
vacuo with
anhydrous THF (4.1 vol.). Repeated azeotrope until specification was met. The
final
solution meeting specifications contained the dissolved crude ER-808227 in
anhydrous THF
(4.1 vol.). Once the specification was met, an appropriately sized inerted
reactor was
charged with anhydrous THF (106 vol.) and the ER-808227 / THE solution from
the
previous step. The reactor was cooled to an internal temperature of -15 to -20
C, then 0.5
M KHMDS in toluene (9.1 wt., 3.0 eq.) was added at a rate such that internal
temperature
did not exceed ¨12 C. Approximately 4.5 eq. KHMDS was necessary to drive the
reaction
to completion. The reaction was reverse quenched into semi-saturated ammonium
chloride
(40 vol.) at an internal temperature of 0 'C. n-Heptane (80 vol.) was added,
stirred for 2-5
minutes, and then allowed to partition. The organic layer was separated, the
aqueous layer
was back extracted with M1BE (70 vol.), then the organic layers were combined
and
washed with saturated sodium chloride solution (70 vol.). The organic layer
was separated
and concentrated in vacuo. To the crude concentrate was added n-heptane (60
vol.). Note:
ER-807363 precipitated out of solution. The resulting suspension was filtered
and the solids
washed with n-heptane (20 vol.). The filtrate was concentrated in vacuo to
afford crude ER-
806746 (-4 wt.) as a brown oil: Note: When additional ER-807363 precipitated
out of
solution, the filtration process was repeated. The crude ER-806746 was
purified via Si02
column chromatography to yield ER-804027 (1.16 wt., 0.55 eq.) as a clear
yellowish oil.
The chromatography was performed as follows: the column was first flushed with
sufficient
MTBE to remove water then flushed with heptane to remove the MTBE. The ER-
806746
84

CA 02822994 2013-08-01
was loaded onto the column as a solution in heptane then eluted from the
column with
heptane/MTBE (5:1) then heptane/M 1BE (4:1) with the fractions monitored at
230 nm by
UV detector.
Me0, Me0,
TBSO ,202Ph TBSO SO2Ph
0 0
0 DIBALH (1 M in CH2Cl2), -78 C
o
CH2O12
oy,OPv I"5or ,OH
ER-804027 ER-804028
[00201] A reactor was charged with ER-804027 (1 wt, 1 eq) and anhydrous
dichloromethane (7.6 vol). The reactor was cooled to an internal temperature
of -78 C and
then I M DIBALH in dichloromethane (3.0 wt, 2.25 eq) was added at a rate such
that
internal temperature did not exceed ¨60 C. Methanol (0.1 vol) was added at a
rate such that
internal temperature did not exceed ¨60 C. Note: hydrogen gas evolved and was
diluted
with a stream of nitrogen. Upon complete addition, the mixture was warmed to
ambient
temperature and then 1 N hydrochloric acid (10.6 vol) and MTBE (25 vol) were
added. The
mixture was stirred for 20 minutes and the layers allowed to partition. The
organic layer
was separated and the aqueous layer back extracted layer with MTBE (15.3 vol).
The
organic layers were combined and washed with water (3 vol), saturated sodium
bicarbonate
(3 vol), and saturated sodium chloride (3 vol), respectively, then
concentrated in vacuo. The
crude concentrate was purified via Si02 column chromatography to yield ER-
804028 (0.84
wt, 0.93 eq) as a white foam.
C. Incorporation of F-3a and Transformations to B-1939:
OTBS
0 11:,1
OTBS
DIBAL-H,
Me0 0 = OTBS Toluene, -75 C
0 OTBS
OTBS
OTBS
ER-803895 ER-803896
[00202] ER-803895 (F-3a) was dissolved in anhydrous toluene (14 wt.) and
cooled to
<-75 C under an argon atmosphere. DIBALH (1.5M in toluene, 0.95 wt., 1.3 eq.)
was

CA 02822994 2013-08-01
added at a rate to maintain the internal reaction temperature <-70 C. The
resulting
mixture was stirred for 30 minutes then quenched with anhydrous methanol (0.13
wt., 3.2
eq.), maintaining the internal reaction temperature <-65 C. The reaction
mixture was
allowed to warm to ¨10 C and transferred with an MTBE rinse (3.74 wt.) to a
workup
vessel containing IN HCI (10.2 wt.). The mixture was stirred for 30 minutes
and the
aqueous layer is drained. The organic phase was washed sequentially with IN 1-
ICI (10.2
wt.), water (10 wt.), saturated aqueous sodium bicarbonate (10 wt.), and brine
(10 wt.) then
concentrated under reduced pressure. The concentrate was purified via silica
gel
chromatography to afford ER-803896 (0.96 wt., 93% yield). The product is
stored at ¨20 C
under argon.
Me0,
TBSO '--rSO2Ph
o OTBS
---- I THF, 0
: OTBS ER-803896
0 I:1
OTBS
ER-804028 ER-803896
SO2Ph
MeO,
TBSO
OH 0 = 0
TBSO OTBS
OTBS
to' ==õ,
0
OH
ER-804029
[00203] At 0 C, a solution of azeotropically dried sulfone ER-804028 (1.0
wt., 1 eq.) in
anhydrous tetrahydrofuran (5 vol., 4.45 wt.) was treated with n-butyl lithium
(1.6M in
hexanes, 1.02 wt., 1.5 vol., 2.05 eq.) such that the internal temperature did
not exceed 5 C.
The mixture was stirred at internal temperature 0 to 5 C for 10 minutes then
cooled to <-75
C. Azeotropically dried aldehyde ER-803896 (1.07wt., 1.23 eq.) was dissolved
in
anhydrous hexanes (3.53 wt., 5_35 vol.) then cooled to <-75 'C. The aldehyde
solution was
added to the ER-804028 anion by cannula such that internal temperature < -65
C. The
mixture was stirred for 45 minutes at internal temperature -78 C then
quenched by the
addition of saturated ammonium chloride (5 vol.), methyl tert-butyl ether (10
vol.), and
water (5 vol.). The aqueous layer was discarded and the organic layer
concentrated under
reduced pressure. The crude material was purified via C-18 reverse phase
chromatography
to afford ER-804029 (84%, 1.57 wt.).
86

CA 02822994 2013-08-01
SO2Ph SO2Ph
_ 0,
TBSO TBSO
0 ,õ0 "=0
TBSO..(0-..

" OH - ' 0
TBSO
OTBS Dess-Main TBSO ATBS
(1)"
TBSO CH2Cl2, rt, 1h TBSO
I
ER-804029 ER-804030
[00204] Sulfone-diol ER-
804029 (1 wt., 1 eq.) was dissolved in wet dichloromethane
(7.4 vol., 0.04 wt% water) and placed in a 20-25 C water bath. Dess-Martin
Reagent (0.67
wt., 2.5 eq.) was added in one portion. The reaction mixture was quenched with
saturated
sodium bicarbonate (10 vol) and 10 wt% aqueous sodium sulfite (10 vol.) and
stirred for 30
minutes. The mixture was diluted with saturated sodium chloride (10 vol) and
extracted
with MTBE (25 vol). The aqueous layer was discarded and the organic layer
concentrated
and purified by silica gel chromatography to afford ER-804030 (0.9 wt., 90%).
The
material was stored under inert gas atmosphere at -20 C.
SO2Ph a
= Q TBSO
TBSOTBSO TBSO,_)c õ 0 0

OTBs SmI2, Me0H, TBSO OTBS
TBSO TBSO
TBSO THF, -78 C
I 0 I
ER-118049
ER-804030
[00205] To a pre-dried
reactor under inert atmosphere was charged samarium diiodide
solution (2.5 eq.) and the solution cooled to internal temperature <-70 'C. ER-
804030 (1
wt.) wa dissolved in anhydrous methanol (4.1 wt.) and anhydrous THF (2.3 wt.)
and then
cooled to <-70 C. ER-804030 was added to the cold samarium solution at a rate
such that
the internal temperature did not exceed ¨70 C. The reaction was quenched with
potassium
carbonate / Rochelle's Salts / water (1/10/100; w/w/v, 15 vol.) and MTBE (5
vol.) such that
internal temperature did not exceed -65 C. Upon complete addition of the
workup solution,
the reaction was warmed to room temperature and the mixture transferred to a
separatory
vessel using the workup solution (20 vol. rinse) and MTBE (20 vol. rinse). The
aqueous
layer was discarded, the organic layer evaporated, and the residue purified
via silica gel
87

CA 02822994 2013-08-01
chromatography to afford ER-118049 (0.77 wt., 85%). The product was stored at
¨20 C
under inert atmosphere.
Me0, Me0,,
TBSO
TBSO TBSO
, 0 0 S TBSO,K, ,.= , 0
0
õ NiCl2, CrCl2, Et3N
()-Ligand ER 807363 0 = 0
OTBS ___________________________________________________________________
OTBS
TBSO
TBSO
OTBS Acetonitrile, THF,
)
30-35 nC TBSO
0
1 0
OH
Keto Aldehyde ER-119049 Allylic
Alcohol ER-118047/048
[00206] A pre-dried reactor was charged with (S)-ligand ER-807363 (2.05 wt)
and the
atmosphere was exchanged for nitrogen. The CrCl2 (0.85 wt, 10 eq) was added in
one
portion followed by anhydrous acetonitrile (21.5 wt) and the mixture was
warmed and
maintained between 30 C to 35 C. Triethylamine (0.7 wt, 0.96 vol, 10 eq) was
added in
one portion and the mixture stirred for one hour. The NiC12(0.09 wt, 1 eq) was
added in one
portion, followed by the keto-aldehyde ER-118049 in anhydrous THF (2.43 wt,
2.73 vol)
over 30 minutes. The heat was removed then heptane (20.5 wt, 30 vol) and
Celite (1.5 wt)
were added. The mixture was stirred for 5 minutes and filtered over a pad of
Celite (15 wt)
and the Celite pad rinsed with heptane (7.3 vol) and acetonitrile (5 vol).
The filtrate was
transferred to a separatory funnel and the lower layer removed. The combined
heptane
layers were washed with acetonitrile (maximum 47.2 wt, maximum 60 vol) as
necessary.
The heptane layer was evaporated under reduced pressure and the product
purified by silica
gel chromatography to afford ER-118047/048 (0.64 wt, 70%).
Me0, Me0
TBSO " TBSO
-, 0 0 0 0
1-1"µ Dess-Martin 0
Hs'
OTBS ____________________________________
TBSO TBSO OTBS
CH2012, RI
TBSO TBSO
0 0
OH 0
ER-118047/048 ER-118046
[002071 Allyl alcohol ER-118047/048 was dissolved in dichloromethane (0.04
wt%
water, 9 vol) and the reactor was placed in a water bath (20 C) and the
solution was treated
with Dess-Martin Reagent (0.48 wt, 1.5 eq). The reaction mixture was treated
with saturated
aqueous sodium bicarbonate (9 vol) and 10 wt% aqueous sodium sulfite (9 vol)
then stirred
88

CA 02822994 2013-08-01
for 20 minutes and transferred to a separatory funnel with DCM (10 vol). The
aqueous
layer was discarded, and the organic layer evaporated to a residue. The crude
material was
purified by flash chromatography (prepped with 3 CV (1:I (WV) DCM/heptane, the

material was loaded with 1:1 DCM/heptane then eluted with 10/10/1
heptane/DCM/IVITBE).
The product-containing fractions were concentrated and stored under inert
atmosphere at ¨
20 C.
Me0_ Me0
TBSO TBSO
TBSOõ.}..õ, , 0 0 H TBSOõ..õ,-c.s., 0
0 0 0
OTBS
O
TBSO TBS TCAA,DIVISO, TBSO
TBSO
Etpl Toluene -15 to 0 G TBSO
0
0
OH 0
ER-118047/048 ER-118046
[00208] Alternatively, the oxidation of ER-118047/48 to form the di-ketone
ER-118046
was accomplished as follows. A flask was charged with ER-118047/48 (1 wt, 1.0
eq) and
toluene (10 vol) and DMSO (0.15 wts, 2.5 eq) were added at room temperature.
Et3N (0.31
wts, 4.0 eq) was added and the solution was cooled to -15 C. TCAA (0.33 wts,
1.4 eq) was
added neat and the reaction warmed to 0 C. then stirred for 10 minutes at 0
C. The
reaction was stirred for additional 10 minutes then was quenched with IPA
(0.15 vol). The
reaction was stirred at 0 C for 10 minutes. IN HCI (5 vol) was added over 2
minutes, and =
the reaction was warmed to room temperature and diluted with MTBE (5 vol). Two
clear
layers formed and the aqueous layer was removed and discarded. The organic
layer was
washed with 5 vol of 5% bicarbonate (aqueous), concentrated to a heavy yellow
oil on a
rotary evaporator and purified by silica gel chromatography (91% isolated
yield).
Me0,
TBSO.
TBSO HO
0 Frs 0 0
0 0 ' 0 "-

OTBS TBAF, THF OH
TBSO HO
TBSO 0
ssõ
0 0
ER 118046 ER 118064
[00209] Into an appropriately sized reaction vessel (vessel A) was charged
imidazole
hydrochloride (0.39 wt, 5 eq) followed by 1 M TBAF in THF (7.6 vol, 10 eq) at
ambient
temperature. The resulting mixture was stirred until it is homogenous (15-30
minutes). Into
a second reaction vessel (vessel B) was charged ER-118046 (1 wt, 1 eq) and THF
(33 vol).
89

CA 02 82 2 9 94 2013-08-01
The contents of vessel B were placed under an inert atmosphere and stirred
until ER-118046
was fully dissolved. The contents of flask A (TBAF/Imidazole) were charged as
a single
portion into flask B (ER-118046/1'HF). After 3-4 days, the reaction solution
was loaded
onto a column and purified by silica gel chromatography.
Me0, Me0,
PPTS, DCM
HO OH Q
0--;=%:;-
õ,,
ER-076349
ER-118064
[002101 The dried ER-118064 (F-12 wherein RI is Me0) residue was dissolved
in
anhydrous dichloromethane (28 vol) under a nitrogen atmosphere and treated
with PPTS
(1.0 wt, 5.2 eq) in one portion. After 30-90 minutes, the reaction mixture was
directly
loaded atop an appropriate column and purified by silica gel chromatography.
The desired
fractions of ER-076349 were concentrated in vacuo . The material resulting
from the
concentration of all pure fractions was azeotroped twice from toluene (20
vol), affording
ER-076349 as a crunchy colorless solid/foam (0.44 wt, 0.79 eq after correction
for residual
toluene).
Me0, Me0,
1. Ts20, Collidine,
Pyridine (cat) " 0
u
0
CH2Cl2, -20 C
,ssoa,
'A)
2. NH4OH, IPA RT0.
0
ER-076349 ER-086526
[002111 In a clean dry reaction vessel (flask C) ER-076349 (1 wt, 1 eq) was
dissolved in
anhydrous toluene (20 vol) and concentrated to dryness under reduced pressure.
The
substrate was re-dissolved in anhydrous toluene (20 vol) and concentrated to
dryness. The
substrate was dissolved in DCM (5 vol), and the solution placed under an argon
atmosphere.
Collidine (0.66 wts, 4.0 eq) was added as a single portion. Pyridine, as a
solution in DCM
(Flask B), was added as a single portion (5 mole %). The resulting mixture in
flask C was
cooled to an internal temperature of ¨20 to -25 C. A DCM solution of Ts20 was
added
drop-wise keeping the internal temperature below ¨16 C (1.02 eq). The
reaction was stirred
at ¨20 to -25 C for 80 minutes then warmed to 0 C over 20 minutes and
stirred for an
additional 20 minutes. The reaction was quenched with water (2 vol). The bath
was

CA 02822994 2013-08-01
removed, and the reaction allowed to warm to room temperature (15-20 C) and
stirred (20
minutes). The reaction was rinsed to a larger vessel using the IPA (100 vol)
and aqueous
ammonium hydroxide (100 vol) was added to the reaction. The reaction was
stirred at room
temperature for 15-36 hours, monitoring for the disappearance of the tosylate
(ER-082892)
and epoxide (ER-809681) which formed in situ. The reaction was concentrated to
dryness or
near dryness at reduced pressure. The resulting material was diluted with DCM
(25-40 vol)
and washed pH 10 buffer (NaHCO3/Na2CO3 (aq), 10 vol). The aqueous phase was
back
extracted with 25 vol of DCM and the combined organic layers were concentrated
to
dryness. The resulting free amine was purified by silica gel chromatography
using a buffered
ACN/water mobile phase. The pooled fractions were concentrated at reduced
pressure to
remove ACN. The resulting aqueous layer was diluted with DCM (40 vol) and with
30 vol
of a pH 10 buffered stock solution (NaHCO3/Na2CO3). The layers were mixed well
and
separated. The aqueous phase was back extracted with 25 vol of DCM and the
combined
organic layers were concentrated to dryness. The resulting free amine was
polish filtered as a
solution in 3:1 DCM/pentane and concentrated to dryness (0.80 wts) to afford B-
1939.
[00212] While we have described a number of embodiments of this invention,
it is
apparent that our basic examples may be altered to provide other embodiments
that utilize
the compounds and methods of this invention. Therefore, it will be appreciated
that the
scope of this invention is to be defined by the appended claims rather than by
the specific
embodiments that have been represented by way of example.
91