Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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HEPATITIS C VIRUS INHIBITORS
FIELD OF THE DISCLOSURE
The present disclosure is generally directed to antiviral compounds, and more
specifically directed to compounds which can inhibit the function of the NS5A
protein encoded by Hepatitis C virus (HCV), compositions comprising such
compounds, and methods for inhibiting the function of the NS5A protein.
BACKGROUND OF THE DISCLOSURE
HCV is a major human pathogen, infecting an estimated 170 million persons
worldwide - roughly five times the number infected by human immunodeficiency
virus type 1. A substantial fraction of these HCV infected individuals develop
serious progressive liver disease, including cirrhosis and hepatocellular
carcinoma.
The current standard of care for HCV, which employs a combination of
pegylated-interferon and ribavirin, has a non-optimal success rate in
achieving
sustained viral response and causes numerous side effects. Thus, there is a
clear and
long-felt need to develop effective therapies to address this undermet medical
need.
HCV is a positive-stranded RNA virus. Based on a comparison of the
deduced amino acid sequence and the extensive similarity in the 5'
untranslated
region, HCV has been classified as a separate genus in the Flaviviridae
family. All
members of the Flaviviridae family have enveloped virions that contain a
positive
stranded RNA genome encoding all known virus-specific proteins via translation
of a
single, uninterrupted, open reading frame.
Considerable heterogeneity is found within the nucleotide and encoded amino
acid sequence throughout the HCV genome due to the high error rate of the
encoded
RNA dependent RNA polymerase which lacks a proof-reading capability. At least
six major genotypes have been characterized, and more than 50 subtypes have
been
described with distribution worldwide. The clinical significance of the
genetic
heterogeneity of HCV has demonstrated a propensity for mutations to arise
during
monotherapy treatment, thus additional treatment options for use are desired.
The
possible modulator effect of genotypes on pathogenesis and therapy remains
elusive.
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The single strand HCV RNA genome is approximately 9500 nucleotides in
length and has a single open reading frame (ORF) encoding a single large
polyprotein of about 3000 amino acids. In infected cells, this polyprotein is
cleaved
at multiple sites by cellular and viral proteases to produce the structural
and non-
structural (NS) proteins. In the case of HCV, the generation of mature non-
structural
proteins (NS2, NS3, NS4A, NS4B, NS5A, and NS5B) is effected by two viral
proteases. The first one is believed to be a metalloprotease and cleaves at
the NS2-
NS3 junction; the second one is a serine protease contained within the N-
terminal
region of NS3 (also referred to herein as NS3 protease) and mediates all the
subsequent cleavages downstream of NS3, both in cis, at the NS3-NS4A cleavage
site, and in trans, for the remaining NS4A-NS4B, NS4B-NS5A, NS5A-NS5B sites.
The NS4A protein appears to serve multiple functions by both acting as a
cofactor for
the NS3 protease and assisting in the membrane localization of NS3 and other
viral
replicase components. The formation of a NS3-NS4A complex is necessary for
proper protease activity resulting in increased proteolytic efficiency of the
cleavage
events. The NS3 protein also exhibits nucleoside triphosphatase and RNA
helicase
activities. NS5B (also referred to herein as HCV polymerase) is a RNA-
dependent
RNA polymerase that is involved in the replication of HCV with other HCV
proteins,
including NS5A, in a replicase complex.
Compounds useful for treating HCV-infected patients are desired which
selectively inhibit HCV viral replication. In particular, compounds which are
effective to inhibit the function of the NS5A protein are desired. The HCV
NS5A
protein is described, for example, in the following references: S. L. Tan, et
al.,
Virology, 284:1-12 (2001); K.-J. Park, et al., J. Biol. Chem., 30711-30718
(2003);
T. L. Tellinghuisen, et al., Nature, 435, 374 (2005); R. A. Love, et al., J.
Virol, 83,
4395 (2009); N. Appel, et al., J. Biol. Chem., 281, 9833 (2006); L. Huang, J.
Biol.
Chem., 280, 36417 (2005); C. Rice, et al., W02006093867.
The present disclosure provides compounds which selectively inhibit HCV
viral replication, as characterized by Formula (I):
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Rq Rv R"' WI'
N...-,....1)--N Rw'
D'1 ,) (. il c2
" HN .-4.1- NH "
z zi
(I),
or a pharmaceutically acceptable salt thereof, wherein:
L is selected from -0-, -CH2CH2 , CH-CH, CC, 0CH2-, -CH20-,
,
e.
I ¨i-
--- ¨I z / A
-CH20CH2-,-% , and
, , ,
X is hydrogen (H) or halogen and Z is hydrogen; or
X and Z, together with the carbon atoms to which they are attached, form a
five- to eight-membered aromatic or non-aromatic fused ring optionally
containing
one or two heteroatoms independently selected from nitrogen, oxygen, and
sulfur;
wherein the five- to eight-membered ring is optionally substituted with one,
two, or
three substitutents independently selected from alkoxy, alkoxyalkyl,
alkoxycarbonyl,
alkyl, alkylsulfonyl, aryl, arylalkyl, arylsulfonyl, carboxy, formyl, halo,
haloalkoxy,
haloalkyl, hydroxy, hydroxyalkyl, -NRaRb, (NRaRb)alkyl, (NRaRb)carbonyl, oxo,
and
spirocycle;
X' is hydrogen (H) or halogen and Z' is hydrogen; or
X' and Z', together with the carbon atoms to which they are attached, form a
five- to eight-membered aromatic or non-aromatic fused ring optionally
containing
one or two heteroatoms independently selected from nitrogen, oxygen, and
sulfur;
wherein the five- to eight-membered ring is optionally substituted with one,
two, or
three substitutents independently selected from alkoxy, alkoxyalkyl,
alkoxycarbonyl,
alkyl, alkylsulfonyl, aryl, arylalkyl, arylsulfonyl, carboxy, formyl, halo,
haloalkoxy,
haloalkyl, hydroxy, hydroxyalkyl, -NRaRb, (NRaRb)alkyl, (NRaRb)carbonyl, oxo,
and
spirocycle;
Y and Y' are each independently -CH2-, -CH2CH2-, or -CH20-, wherein the
-CH20- is drawn such that the oxygen atom is bound to the carbon atom
substituted
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with Rv and Rq or R"' and WI;
RP is hydrogen or Ci to C4 alkyl;
Rq is hydrogen, alkyl, or halo; or
RP and Rq, together with the carbon atoms to which they are attached, form a
cycloalkyl ring;
Rv is selected from hydrogen, alkyl, halo, and hydroxy; or
Rv and Rq, together with the carbon atom to which they are attached, form an
ethylenyl group or a cycloalkyl ring;
RP' is hydrogen or Ci to C4 alkyl;
Rq' is hydrogen, alkyl, or halo; or
RP' and Rq', together with the carbon atoms to which they are attached, form a
cycloalkyl ring;
R'' are independently selected from hydrogen, alkyl, halo, and hydroxy; or
R"' and WI, together with the carbon atom to which they are attached, form an
ethylenyl group or a cycloalkyl ring;
Rw and Rw' are independently selected from hydrogen and alkyl;
R1 is hydrogen or
R2 is hydrogen or
Rx and RY are independently selected from cycloalkyl, heteroaryl,
heterocyclyl, alkoxy, and alkyl, said alkyl being substituted by one or more
substituents independently selected from aryl, alkenyl, cycloalkyl,
heterocyclyl,
heteroaryl, -0R3, -C(0)0R4, -NRaRb, and -C(0)NReRd,
wherein any said aryl and heteroaryl may optionally be substituted with one
or more substituents independently selected from alkenyl, alkyl, haloalkyl,
arylalkyl,
heterocyclyl, heterocyclylalkyl, halogen, cyano, nitro, -C(0)0R4, -0R5, -
NRaRb,
(NRaRb)alkyl, and (Me0)(HO)P(0)0-, and
wherein any said cycloalkyl and heterocyclyl may optionally be fused onto an
aromatic ring and may optionally be substituted with one or more substituents
independently selected from alkyl, hydroxyl, halogen, aryl, -NRaRb, oxo, and
-C(0)0R4;
R3 is hydrogen, alkyl, or arylalkyl;
R4 is alkyl or arylalkyl;
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R5 is hydrogen, alkyl, or arylalkyl;
Ra and Rb are independently selected from hydrogen, alkyl, cycloalkyl,
arylalkyl, heteroaryl, -C(0)R6, -C(0)0R7, ¨C(0)NReRd, and (NReRd)alkyl, or
alternatively, Ra and Rb, together with the nitrogen atom to which they are
attached,
form a five- or six-membered ring or bridged bicyclic ring structure, wherein
said
five- or six-membered ring or bridged bicyclic ring structure optionally may
contain
one or two additional heteroatoms independently selected from nitrogen,
oxygen, and
sulfur and may contain one, two, or three substituents independently selected
from Ci
to C6 alkyl, C1 to C4 haloalkyl, aryl, hydroxyl, Ci to C6 alkoxy, Ci to C4
haloalkoxy,
and halogen;
R6 is alkyl;
R7 is alkyl, arylalkyl, cycloalkyl, or haloalkyl; and
Re and Rd are independently selected from hydrogen, alkyl, arylalkyl, and
cycloalkyl.
In a first embodiment of the first aspect the present disclosure provides a
compound of Formula (I) further characterized by Formula (Ia):
N Rv
4 ,.........-- WI
N \ N
N H I L H N
RP R1 /. R2 RP
Z ZI
(Ia),
or a pharmaceutically acceptable salt or a tautomer thereof, wherein:
X is hydrogen or chloro (Cl) and Z is hydrogen; or
X and Z, together with the carbon atoms to which they are attached, form a
six-membered aromatic or non-aromatic fused ring;
X' is hydrogen or chloro (Cl) and Z' is hydrogen; or
X' and Z', together with the carbon atoms to which they are attached, form a
six-membered aromatic or non-aromatic fused ring;
Y is -CH2-, -CH2CH2-, or -CH20-, wherein the -CH20- is drawn such that the
oxygen atom is bound to the carbon atom substituted with Rv and Rq;
RP is hydrogen or C1 to C4 alkyl;
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Rq is hydrogen, alkyl, or halo; or
RP and Rq, together with the carbon atoms to which they are attached, form a
cycloalkyll ring; and
Rv is selected from hydrogen, alkyl, halo, and hydroxy; or
Rv and Rq, together with the carbon atom to which they are attached, form an
ethylenyl group or a cycloalkyl ring.
In a second embodiment of the first aspect the present disclosure provides a
compound of Formula (I) further characterized by Formula (Ib):
X X'
N N
N N
(Ib),
or a pharmaceutically acceptable salt or a tautomer thereof
In a third embodiment of the first aspect the present disclosure provides a
compound of Formula (I) further characterized by Formula (Ic):
X X'
N N
N N
H H3 ¨L H N
3C C
/ / R2
IR 1
(Ic),
or a pharmaceutically acceptable salt or a tautomer thereof
In a fourth embodiment of the first aspect the present disclosure provides a
compound of Formula (I) further characterized by Formula (Id):
X X'
N N
N N
(Id),
or a pharmaceutically acceptable salt or a tautomer thereof
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In a fifth embodiment of the first aspect the present disclosure provides a
compound of Formula (Ia), or a pharmaceutically acceptable salt thereof,
wherein:
R1 is _C(0)R';
R2 is ¨C(0)R";
Rx and RY are independently alkyl substituted by at least one -NRaRb,
characterized by Formula (A):
R Rb
a, N.
R8---A ________________________________ )3-
R9
(A),
wherein:
m is 0 or 1;
R8 is hydrogen or alkyl;
R9 is selected from hydrogen, cycloalkyl, aryl, heteroaryl, heterocyclyl, and
alkyl optionally substituted with a substituent selected from aryl, alkenyl,
cycloalkyl,
heterocyclyl, heteroaryl, heterobicyclyl, -0R3, -C(0)0R4, -NRaRb, and -
C(0)NReRd,
wherein any said aryl and heteroaryl may optionally be substituted with one or
more
substituents independently selected from alkyl, haloalkyl, arylalkyl,
heterocyclyl,
heterocyclylalkyl, halogen, cyano, nitro, -C(0)0R4, -0R5, -NRaRb,
(NRaRb)alkyl,
and (Me0)(HO)P(0)0-, and
wherein any said cycloalkyl and heterocyclyl may optionally be fused onto an
aromatic ring and may optionally be substituted with one or more substituents
independently selected from alkyl, hydroxyl, halogen, aryl, -NRaRb, oxo, and
-C(0)0R4; and
R3, R4, R5, Ra, Rb, Re, and Rd are defined as in Formula (I).
In a sixth embodiment of the first aspect the present disclosure provides a
compound of Formula (Ia) or a pharmaceutically acceptable salt thereof,
wherein
m is 0;
R8 is hydrogen or Ci to C4 alkyl;
R9 is selected from hydrogen, Ci to C6 alkyl optionally substituted with ¨
OR12, C3 to C6 cycloalkyl, allyl, -CH2C(0)NReRd, (NReRd)alkyl,
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(R10)n (R10)n
(Rio)n (r,10
?4,
N> j SN g\N
\ 10
(R10) cryT (R )n (R10)n
L,N
N I ) I tkirss,3
S
(R10 n (R10)n (R10 n
4Vr'l< /4-
0 N
(R10)n
iol=10\
(Rio)n krx )n
ol LK" N
; !.
\
R11
,and
(Rlo)n N
wherein j is 0 or 1;
k is 1, 2, or 3;
n is 0 or an integer selected from 1 through 4;
each R1 is independently hydrogen, Ci to C4 alkyl, Ci to C4 haloalkyl,
halogen, nitro, -0Bn, or (Me0)(OH)P(0)0-;
R11 is hydrogen, Ci to C4 alkyl, or benzyl;
R12 is hydrogen, Ci to C4 alkyl, or benzyl;
Ra is hydrogen or Ci to C4 alkyl;
Rb is C1 to C4 alkyl, C3 to C6 cycloalkyl, benzyl, 3-pyridyl, pyrimidin-5-yl,
acetyl, -C(0)0R7, or -C(0)NReRd;
R7 is Ci to C4 alkyl or Ci to C4 haloalkyl;
Re is hydrogen or C1 to C4 alkyl; and
Rd is hydrogen, Ci to C4 alkyl, or C3 to C6 cycloalkyl.
In a seventh embodiment of the first aspect the present disclosure provides a
compound of Formula (Ia) or a pharmaceutically acceptable salt thereof,
wherein
m is 0;
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R8 is hydrogen;
R9 is phenyl optionally substituted with one up to five substituents
independently selected from Ci to C6 alkyl, Ci to C4 haloalkyl, halogen, C1 to
C6
alkoxy, hydroxyl, cyano, and nitro; and
NRaRb is a heterocyclyl or heterobicyclyl group selected from:
(R13), (R13),
-EN\_ /1_/-\
N¨R14 1-N 0
,
,
,
1
N
41(w (R13)nx9....,,
N
C
47.¨(R13)n N
R14
,and ,
wherein n is 0, 1, or 2;
each R13 is independently selected from Ci to C6 alkyl, phenyl,
trifluoromethyl, halogen, hydroxyl, methoxy, and oxo; and
R14 is C1
to C6 alkyl, phenyl, benzyl, or -C(0)0R15 group, wherein R15 is Ci
to C4 alkyl, phenyl, or benzyl.
In an eighth embodiment of the first aspect the present disclosure provides a
compound of Formula (Ia) or a pharmaceutically acceptable salt thereof,
wherein
m is 1;
R8 is hydrogen;
R9 is Ci to C6 alkyl, arylalkyl, or heteroarylalkyl;
Ra is hydrogen; and
Rb is ¨C(0)0R7, wherein R7 is Ci to C6 alkyl.
In a ninth embodiment of the first aspect the present disclosure provides a
compound of Formula (Ia) or a pharmaceutically acceptable salt thereof,
wherein
R1 is _C(0)R';
R2 is ¨C(0)R";
Rx and RY are heteroaryl or heterocyclyl independently selected from:
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(R13)n (R13)n (R13)n \(R13
1
) \ , I
4I 24.= 4n
n 1
Cµ-
N)
1 1
(R13) vivs, ,
n. =^-
"\-...._N R 13
N '"1N \ 1 )n
R14
/ / /
I --1- in IN (R13)n T
S 114 R14 R14-NN/ / / /
I I (R13 )n
., v
(R13)
I (R13)n I N¨r?zi
,\:--_, F\ __:--M
R ,N, ...j..) S N)
N N
U N
, , , ,
(R13)n R13
(R13)fl
ciN-R14 (III R14
,
I
\ ) Ty
N 1
, and
-'*----.../ -14 R14
, R , ,
wherein n is 0 or an integer selected from 1 through 4;
each R13 is independently selected from hydrogen, Ci to C6 alkyl, Ci to C4
haloalkyl, phenyl, benzyl, Ci to C6 alkoxy, Ci to C4 haloalkoxy, heterocyclyl,
halogen, NIZeRd, hydroxyl, cyano, and oxo, where Re and Rd are independently
hydrogen or Ci to C4 alkyl; and
R14 is hydrogen (H), C1 to C6 alkyl, benzyl, or ¨C(0)0R4, wherein R4 is C1 to
C6 alkyl.
In a tenth embodiment of the first aspect the present disclosure provides a
compound of Formula (Ia) or a pharmaceutically acceptable salt thereof,
wherein
R1 is _C(0)R';
R2 is ¨C(0)R";
Rx and RY are cycloalkyl independently selected from:
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NRaRb
(R13) NRaRb,
and
wherein
j is 0, 1, 2, or 3;
k is 0, 1, or 2;
n is 0 or an integer selected from 1 through 4;
each R13 is independently selected from hydrogen, Ci to C6 alkyl, Ci to C4
haloalkyl, Ci to C6 alkoxy, halogen, hydroxyl, cyano, and nitro; and
Ra and Rb are each independently hydrogen, Ci to C6 alkyl, or -C(0)0R7,
wherein R7 is C1 to C6 alkyl.
In an eleventh embodiment of the first aspect the present disclosure provides
a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof,
wherein
R1 is _C(0)R';
R2 is ¨C(0)R";
Rx and RY are independently arylalkyl, wherein aryl part of said arylalkyl may
optionally be substituted with (NRaRb)alkyl; and
Ra and Rb are independently hydrogen, Ci to C6 alkyl, or benzyl, or
alternatively, Ra and Rb, together with the nitrogen atom to which they are
attached,
form a five-or six-membered ring selected from =Fd
, and , wherein R15 is hydrogen, Ci to C6 alkyl, or
benzyl.
In a twelfth embodiment of the first aspect the present disclosure provides a
compound of Formula (Ia) or a pharmaceutically acceptable salt thereof,
wherein
R1 and R2 are the same and are selected from the group consisting of:
tei
0
H 0 H C)11
HN0 OyN ssss, oy Ntsss,, ()yN
NH C NH2
0 0 r)i-2 )1-4 __ 0
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H j?
0
y
0
y555-s 0
0 0 N H
0
0)'LNH
0 411 0
NN3/yLs.se
OH
Y
HN 0 Os n
'22 ......õ.,
P
Ph----/ 0 , Me0- \
OH 0 0
,and
,
Ck /
\¨NH 0
0
. 0
*Pak0 NH
IIW riif
/
wherein a squiggle bond (¨ ) in the structure indicates that a stereogenic
center to
which the bond is attached can take either (R)- or (S)- configuration so long
as
chemical bonding principles are not violated.
In a thirteenth embodiment of the first aspect the present disclosure provides
a
compound of Formula (I), or a pharmaceutically acceptable salt thereof,
wherein
R1 is _C(0)R';
10R2 =
is ¨C(0)R'; and
Rx and RY are both t-butoxy.
In a fourteenth embodiment of the first aspect the present disclosure provides
a compound of Formula (I), or a pharmaceutically acceptable salt thereof,
wherein
R1 and R2 are both hydrogen.
In a second aspect the present disclosure provides a compound of Formula
(II):
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Rq\._ Rcr
RP¨------- ------¨RP'
N
R1 FIN,I,, ...1,NEI
R2
1
x x,
(I),
or a pharmaceutically acceptable salt thereof, wherein:
L is selected from ¨0¨, ¨CH2CH2 , CH¨CH, CC, OCH2¨, -CH20¨,
e.
A
1-..õ..) .-- ,... A..
-CH2OCH2¨, , and ,
X and X' are independently hydrogen (H) or halogen;
RP is hydrogen or C1 to C4 alkyl, and Rq is hydrogen, or alternatively, RP and
Rq, together with the carbon atoms to which they are attached, form a
cyclopropyl
ring;
RP' is hydrogen or C1 to C4 alkyl, and Rq' is hydrogen, or alternatively, RP'
and Rq', together with the carbon atoms to which they are attached, form a
cyclopropyl ring;
R1 is hydrogen or
R2 is hydrogen or
Rx and RY are independently selected from cycloalkyl, heteroaryl,
heterocyclyl, alkoxy, and alkyl, said alkyl being substituted by one or more
substituents independently selected from aryl, alkenyl, cycloalkyl,
heterocyclyl,
heteroaryl, -0R3, -C(0)0R4, -NRaRb, and -C(0)NReRd,
wherein any said aryl and heteroaryl may optionally be substituted with one
or more substituents independently selected from alkyl, haloalkyl, arylalkyl,
heterocyclyl, heterocyclylalkyl, halogen, cyano, nitro, -C(0)0R4, -0R5, -
NRaRb,
(NRaRb)alkyl, and (Me0)(HO)P(0)0-, and
wherein any said cycloalkyl and heterocyclyl may optionally be fused onto an
aromatic ring and may optionally be substituted with one or more substituents
independently selected from alkyl, hydroxyl, halogen, aryl, -NRaRb, oxo, and
-C(0)0R4;
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R3 is hydrogen, alkyl, or arylalkyl;
R4 is alkyl or arylalkyl;
R5 is hydrogen, alkyl, or arylalkyl;
Ra and Rb are independently selected from hydrogen, alkyl, cycloalkyl,
arylalkyl, heteroaryl, -C(0)R6, -C(0)0R7, ¨C(0)NReRd, and (NReRd)alkyl, or
alternatively, Ra and Rb, together with the nitrogen atom to which they are
attached,
form a five- or six-membered ring or bridged bicyclic ring structure, wherein
said
five- or six-membered ring or bridged bicyclic ring structure optionally may
contain
one or two additional heteroatoms independently selected from nitrogen,
oxygen, and
sulfur and may contain one, two, or three substituents independently selected
from Ci
to C6 alkyl, C1 to C4 haloalkyl, aryl, hydroxyl, Ci to C6 alkoxy, Ci to C4
haloalkoxy,
and halogen;
R6 is alkyl;
R7 is alkyl, arylalkyl, or haloalkyl; and
Re and Rd are independently selected from hydrogen, alkyl, arylalkyl, and
cycloalkyl.
In a third aspect the present disclosure provides a composition comprising a
compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, wherein Formula (I) is defined according
to any
of the embodiments described above in the first aspect of the present
disclosure.
In a first embodiment of the third aspect the composition further comprises at
least one additional compound having anti-HCV activity.
In a second embodiment of the third aspect at least one of the additional
compounds is an interferon or a ribavirin.
In a third embodiment of the third aspect the interferon is selected from
interferon alpha 2B, pegylated interferon alpha, consensus interferon,
interferon
alpha 2A, and lymphoblastiod interferon tau.
In a fourth embodiment of the third aspect the present disclosure provides a
composition comprising a compound of Formula (I), or a pharmaceutically
acceptable salt thereof, a pharmaceutically acceptable carrier, and at least
one
additional compound having anti-HCV activity, wherein at least one of the
additional
compounds is selected from interleukin 2, interleukin 6, interleukin 12, a
compound
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that enhances the development of a type 1 helper T cell response, interfering
RNA,
anti-sense RNA, Imiqimod, ribavirin, an inosine 5'-monophospate dehydrogenase
inhibitor, amantadine, and rimantadine.
In a fifth embodiment of the third aspect the present disclosure provides a
composition comprising a compound of Formula (I), or a pharmaceutically
acceptable salt thereof, a pharmaceutically acceptable carrier, and at least
one
additional compound having anti-HCV activity, wherein at least one of the
additional
compounds is effective to inhibit the function of a target selected from HCV
metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B
protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, and IMPDH
for the treatment of an HCV infection.
In a fourth aspect the present disclosure provides a method of treating an
HCV infection in a patient, comprising administering to the patient a
therapeutically
effective amount of a compound of Formula (I), or a pharmaceutically
acceptable salt
thereof, wherein Formula (I) is defined according to any of the embodiments
described above in the first aspect of the present disclosure.
In a first embodiment of the fourth aspect the method further comprises
administering at least one additional compound having anti-HCV activity prior
to,
after or simultaneously with the compound of Formula (I), or a
pharmaceutically
acceptable salt thereof
In a second embodiment of the fourth aspect at least one of the additional
compounds is an interferon or a ribavirin.
In a third embodiment of the fourth aspect the interferon is selected from
interferon alpha 2B, pegylated interferon alpha, consensus interferon,
interferon
alpha 2A, and lymphoblastiod interferon tau.
In a fourth embodiment of the fourth aspect the present disclosure provides a
method of treating an HCV infection in a patient, comprising administering to
the
patient a therapeutically effective amount of a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, and at least one additional compound
having anti-HCV activity prior to, after or simultaneously with the compound
of
Formula (I), or a pharmaceutically acceptable salt thereof, wherein at least
one of the
additional compounds is selected from interleukin 2, interleukin 6,
interleukin 12, a
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compound that enhances the development of a type 1 helper T cell response,
interfering RNA, anti-sense RNA, Imiqimod, ribavirin, an inosine 5'-
monophospate
dehydrogenase inhibitor, amantadine, and rimantadine.
In a fifth embodiment of the fourth aspect the present disclosure provides a
method of treating an HCV infection in a patient, comprising administering to
the
patient a therapeutically effective amount of a compound of Formula (I), or a
pharmaceutically acceptable salt thereof, and at least one additional compound
having anti-HCV activity prior to, after or simultaneously with the compound
of
Formula (I), or a pharmaceutically acceptable salt thereof, wherein at least
one of the
additional compounds is effective to inhibit the function of a target selected
from
HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV
NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, and
IMPDH for the treatment of an HCV infection.
The compounds of the present disclosure can be effective to inhibit the
function of the HCV NS5A protein. In particular, the compounds of the present
disclosure can be effective to inhibit the HCV lb genotype or multiple
genotypes of
HCV. Therefore, this disclosure also encompasses: (1) compositions comprising
a
compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier; and (2) a method of treating an HCV
infection in
a patient, comprising administering to the patient a therapeutically effective
amount
of a compound of Formula (I), or a pharmaceutically acceptable salt thereof
Other aspects of the present disclosure may include suitable combinations of
embodiments disclosed herein.
Yet other aspects and embodiments may be found in the description provided
herein.
The description of the present disclosure herein should be construed in
congruity with the laws and principals of chemical bonding. In some instances
it
may be necessary to remove a hydrogen atom in order to accommodate a
substituent
at any given location.
Certain features of the structure of Formula (I) are further illustrated
below:
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Rq Rv yRv,...'ccl' .
RP7 Y1.--
RP
N
RA 'CrN\ N-..,õ(1¨N Rw
1 7
R'l NH R-
,..._,
I I
Z'
Pyrrolidinyl-imidazole
Pyrrolidinyl-imidazole
(Left) Linker (Right)
(I).
In Formula (I), as depicted above, the "pyrrolidinyl-imidazole" moiety on the
left side of the "linker" is independent from the "pyrrolidinyl-imidazole"
moiety on
the right side of the linker group in respect to, e.g., (1) tautomer form of
imidazole
ring, (2) absolute configuration of the stereogenic centers on the pyrrolidine
ring, and
(3) substituents on the pyrrolidine nitrogen, i.e., R1 and R2 are independent
from each
other, although in some circumstances they are preferably the same.
It should be understood that the depiction of a pyrrolidine moiety on the
"left"
side or on the "right" side is for illustration purpose only, which does not
in any way
limit the scope of the disclosure.
In the linker group of Formula (I), the linkage between "L" and the two
benzene rings encompasses all the following combinations:
;sss = or 'zza,, *
I. 1'
2 4 2' 2
L L ss's
(para-para), (para-meta),
;cps'
:?.?õ =
..,01 3 L 3' 0 1sr's ' ..3 2 4 L2' I.
3 1.
2 2'
(meta-meta), 'I' (para-ortho),
0 2 3' I. 10 2 2' 10 L .rsa 1 L 1.
2'
I (meta-ortho), and I . (ortho-ortho),
wherein the "para-para," "para-meta," and "meta-meta" linkages are preferred.
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k k
Likewise, in Formula (I), when L is a phenylene (--- ) group, it can
link to the adjacent two benzene rings by the following manners:
2 3
1/\2 1.4 /-
(para), 1_(meta), and W
(ortho),
wherein the "para" and "meta" arrangements are preferred, and the "para"
arrangement is the more preferred.
In Formula (I), when L is a vinylene (-CH=CH-) group, it can take either
trans- or cis- configuration, as depicted below:
k_
k 1
(cis-), (trans-).
NA
In Formula (I), when L is a cyclopropylene (.,... A) group, the two
benzene substituents can be either trans- or cis- to each other, forming one
of the
following four configurations:
A
00"1\410õ 5 _i_
- e....,,o. .,õ..,.........õ, 1 1 _..- 1_ 1
Ji -1-
...., ...
(cis-), (cis-),
71¨
.
-...
(trans-),(trans-).
1.....,s,
In a pyrrolidine ring of a pyrrolidinyl-imidazole moiety, the stereogenic
carbon center to which the imidazole group is attached can take either (R)- or
(S)-
configuration as depicted below:
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Ry
Rq N
Ricv
RP Rw
Rw
(R-) (S-)
When a cyclopropyl ring is fused onto a pyrrolidine ring of a pyrrolidinyl-
imidazole moiety, i.e., when (RP, Rq) together is ¨CH2-, the CH2 group of the
fused
cyclopropyl ring can take either a- or 13- position relative to the
pyrrolidine ring, as
depicted below:
N--,
N-11
_2) <
R1 (PA R1
Thus, this disclosure is intended to cover all possible stereoisomers even
when a single stereoisomer, or no stereochemistry, is described in a
structure.
In Formula (I), the linkage between a benzene ring of the linker group and an
imidazole ring of a pyrrolidinyl-imidazole moiety can take place in either the
C-4 or
the C-5 position (see below) of the imidazole ring. As a person of ordinary
skill in
the art would understand, due to tautomerization of the imidazole ring, a
bonding of a
benzene ring to the C-4 position may be equivalent to a bonding of the benzene
ring
to the C-5 position, as shown in the following equation:
R11 R1
R 3
NI 3 N-Th 4
tautomerization HN
______________________________ r r
N 5.sss
The sample principle also applies to substituent X or X'.
Thus, this disclosure is intended to cover all possible tautomers even when a
structure depicts only one of them.
In this disclosure, a floating bond (e.g., ) or a floating substituent
(e.g.,
-R13) on a structure indicates that the bond or substituent can attach to any
available
position of the structure by removal of a hydrogen from the available
position. It
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should be understood that in a bicyclic or polycyclic ring structure, unless
specifically defined otherwise, the position of a floating bond or a floating
substituent
does not limit the position of such bond or substituent to a specific ring.
Thus, the
following two substituents should be construed to be equivalent:
(R13),,
(R13)
/--0 and 0 .
It should be understood that the compounds encompassed by the present
disclosure are those that are suitably stable for use as pharmaceutical agent.
It is intended that the definition of any substituent or variable at a
particular
location in a molecule be independent of its definitions elsewhere in that
molecule.
For example, for substituent (R10)õ, when n is 2, each of the two R1 groups
may be
the same or different.
All patents, patent applications, and literature references cited in the
specification are herein incorporated by reference in their entirety. In the
case of
inconsistencies, the present disclosure, including definitions, will prevail.
DEFINITIONS
Definitions have been provided above for each of the groups defined. In
addition, the following definitions shall be used.
As used herein, the singular forms "a", "an", and "the" include plural
reference unless the context clearly dictates otherwise.
Unless stated otherwise, all aryl, cycloalkyl, heteroaryl, and heterocyclyl
groups of the present disclosure may be substituted as described in each of
their
respective definitions. For example, the aryl part of an arylalkyl group may
be
substituted as described in the definition of the term "aryl."
The term "acetyl," as used herein, refers to ¨C(0)CH3.
The term "alkenyl," as used herein, refers to a monovalent, straight or
branched hydrocarbon chain having one or more, preferably one to two, double
bonds therein. The double bond of an alkenyl group can be unconjugated or
conjugated to another unsaturated group. Suitable alkenyl groups include, but
are not
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limited to, C2 to C10 alkenyl groups, such as vinyl, allyl, butenyl, pentenyl,
hexenyl,
butadienyl, pentadienyl, hexadienyl, 2-ethylhexenyl, 2-propy1-2-butenyl, 4-(2-
methy1-3-butene)-pentenyl. An alkenyl group can be unsubstituted or
substituted
with one or two suitable substituents.
The term "alkoxy," as used herein, refers to an alkyl group attached to the
parent molecular moiety through an oxygen atom. Representative examples of
alkoxy group include, but are not limited to, methoxy (CH30-), ethoxy (CH3CH20-
),
and t-butoxy ((CH3)3C0-).
The term "alkoxyalkyl," as used herein, refers to an alkyl group substituted
with one, two, or three alkoxy groups.
The term "alkoxycarbonyl," as used herein, refers to an alkoxy group attached
to the parent molecular moiety through a carbonyl group.
The term "alkyl," as used herein, refers to a group derived from a straight or
branched chain saturated hydrocarbon by removal of a hydrogen from one of the
saturated carbons. The alkyl group preferably contains from one to ten carbon
atoms.
Representative examples of alkyl group include, but are not limited to,
methyl, ethyl, isopropyl, and tert-butyl.
The term "alkylcarbonyl," as used herein, refers to an alkyl group attached to
the parent molecular moiety through a carbonyl group. Representative examples
of
alkylcarbonyl group include, but are not limited to, acetyl (-C(0)CH3),
propanoyl
(-C(0)CH2CH3), n-butyryl (-C(0)CH2CH2CH3), and 2,2-dimethylpropanoyl or
pivaloyl (-C(0)C(CH3)3).
The term "alkylsulfonyl," as used herein, refers to an alkyl group attached to
the parent molecular moiety through a sulfonyl group.
The term "allyl," as used herein, refers to the ¨CH2CH=CH2 group.
The term "aryl," as used herein, refers to a group derived from an aromatic
carbocycle by removal of a hydrogen atom from an aromatic ring. The aryl group
can be monocyclic, bicyclic or polycyclic, wherein in bicyclic or polycyclic
aryl
group, the aromatic carbocycle can be fused onto another four- to six-membered
aromatic or non-aromatic carbocycle. Representative examples of aryl groups
include, but are not limited to, phenyl, indanyl, indenyl, naphthyl, and
1,2,3,4-
tetrahydronaphth-5-yl.
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The term "arylalkyl," as used herein, refers to an alkyl group substituted
with one,
two, or three aryl groups, wherein aryl part of the arylalkyl group may
optionally be
substituted by one to five substituents independently selected from Ci to C6
alkyl, Ci
to C4 haloalkyl, Ci to C6 alkoxy, halogen, cyano, and nitro groups.
Represented
examples of arylalkyl include, but are not limited to, benzyl, 2-phenyl-1-
ethyl
(PhCH2CH2-), (naphth-l-yl)methyl, and (naphth-2-yl)methyl.
The term "arylsulfonyl," as used herein, refers to an aryl group attached to
the
parent molecular moiety through a sulfonyl group.
The term "benzyl," as used herein, refers to a methyl group on which one of
the hydrogen atoms is replaced by a phenyl group, wherein said phenyl group
may
optionally be substituted by one to five substituents independently selected
from
methyl, trifluoromethyl (-CF3), methoxy (-0CH3), halogen, and nitro (-NO2).
Representative examples of benzyl group include, but are not limited to,
PhCH2-, 4-Me0-C6H4CH2-, and 2,4,6-tri-methyl-C6H4CH2-.
The term "bridged bicyclic ring," as used herein, refers to a ring structure
comprising a bridgehead between two of the ring members, wherein the ring and
the
bridgehead optionally may independently comprise one or more, preferably one
to
two, heteroatoms independently selected from nitrogen, oxygen, and sulfur.
Illustrated examples of a bridged bicyclic ring structure include, but are not
limited
to:
I
siV".
I
vz=uv N (()......\.õN
1
N
, ,and RN .
The terms "Cap" and "cap," as used herein, refer to the group which is placed
on the nitrogen atom of the pyrrolidine ring in the compounds of formula (I).
It
should be understood that "Cap" or "cap" can also refer to the reagent which
is a
precursor to the final "cap" in compounds of formula (I) and is used as one of
the
starting materials in the reaction to append a group on the pyrrolidine
nitrogen that
results in the final product, a compound which contains the functionalized
pyrrolidine
that will be present in the compound of formula (I).
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The term "carbonyl," as used herein, refers to -C(0)-.
The term "carboxyl," or "carboxy," as used herein, refers to -CO2H.
The term "cyano," as used herein, refers to -CN.
The term "cycloalkyl," as used herein, refers to a group derived from a
saturated carbocycle, having preferably three to eight carbon atoms, by
removal of a
hydrogen atom from the saturated carbocycle, wherein the saturated carbocycle
can
optionally be fused onto one or two other aromatic or nonaromatic carbocycles.
Representative examples of cycloalkyl groups include, but are not limited to,
cyclopropyl, cyclopentyl, cyclohexyl, and 1,2,3,4-tetrahydronaphth-1-yl.
The term "formyl," as used herein, refers to -CHO.
The terms "halo" and "halogen," as used herein, refer to F, Cl, Br, or I.
The term "haloalkoxy," as used herein, refers to a haloalkyl group attached to
the
parent molecular moiety through an oxygen atom.
The term "haloalkyl," as used herein, refers to an alkyl group substituted by
at
least one halogen atom. The haloalkyl group can be an alkyl group of which all
hydrogen atoms are substituted by halogens. Representative examples of
haloalkyl
include, but are not limited to, trifluoromethyl (CF3-), 1-chloroethyl
(C1CH2CF127),
and 2,2,2-trifluoroethyl (CF3CH2-).
The term "heteroaryl," as used herein, refers to group derived from a
monocyclic, bicyclic, or polycyclic compound comprising at least one aromatic
ring
comprising one or more, preferably one to three, heteroatoms independently
selected
from nitrogen, oxygen, and sulfur, by removal of a hydrogen atom from an
aromatic
ring thereof As is well known to those skilled in the art, heteroaryl rings
have less
aromatic character than their all-carbon counterparts. Thus, for the purposes
of the
disclosure, a heteroaryl group need only have some degree of aromatic
character.
Illustrative examples of heteroaryl groups include, but are not limited to,
pyridyl,
pyridazinyl, pyrimidyl, pyrazyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl,
(1,2,3,)-
and (1,2,4)-triazolyl, pyrazinyl, pyrimidinyl, tetrazolyl, furyl, thienyl,
isoxazolyl,
thiazolyl, isoxazolyl, oxazolyl, indolyl, quinolinyl, isoquinolinyl,
benzisoxazolyl,
benzothiazolyl, benzothienyl, and pyrrolopyridinyl.
The term "heteroarylalkyl," as used herein, refers to an alkyl group
substituted with one, two, or three heteroaryl groups.
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The term "heterobicyclyl," as used herein, refers to a ring structure
comprising two fused or bridged rings that include carbon and one or more,
preferably one to three, heteroatoms independently selected from nitrogen,
oxygen,
and sulfur. The heterobicyclic ring structure is a subset of heterocyclic ring
and can
be saturated or unsaturated. Examples of heterobicyclic ring structures
include, but
are not limited to, tropane, quinuclidine, and 7-azabicyclo[2.2.1]heptane.
The term "heterocyclyl," as used herein, refers to a group derived from a
monocyclic, bicyclic, or polycyclic compound comprising at least one
nonaromatic
ring comprising one or more, preferably one to three, heteroatoms
independently
selected from nitrogen, oxygen, and sulfur, by removal of a hydrogen atom from
the
nonaromatic ring. The heterocyclyl group encompasses the heterobicyclyl group.
The heterocyclyl groups of the present disclosure can be attached to the
parent
molecular moiety through a carbon atom or a nitrogen atom in the group.
Examples
of heterocyclyl groups include, but are not limited to, morpholinyl,
oxazolidinyl,
piperazinyl, piperidinyl, pyrrolidinyl, tetrahydrofuryl, thiomorpholinyl, and
indolinyl.
The term "heterocyclylalkyl," as used herein, refers to an alkyl group
substituted with one, two, or three heterocyclyl groups.
The terms "hydroxy" or "hydroxyl," as used herein, refer to -OH.
The term "hydroxyalkyl," as used herein, refers to an alkyl group substituted
with one, two, or three hydroxy groups.
The term "nitro," as used herein, refers to -NO2.
The term "-NRaRb," as used herein, refers to two groups, Ra and Rb, which are
attached to the parent molecular moiety through a nitrogen atom, or
alternatively Ra
and Rb, together with the nitrogen atom to which they are attached, form a 5-
or 6-
membered ring or a fused- or bridged-bicyclic ring structure optionally
containing
one, two, or three additional heteroatom independently selected from nitrogen,
oxygen, and sulfur. The term "-NReRd" is defined similarly.
The term "(NRaRb)alkyl," as used herein, refers to an alkyl group substituted
with one, two, or three ¨NRaRb groups. The term "(NReRd)alkyl" is defined
similarly.
The term "oxo," as used herein, refers to =0.
The term "sulfonyl," as used herein, refers to -SO2-.
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The term "trialkylsilyl," as used herein, refers to -SiR3, wherein each R is
C1
to C4 alkyl or phenyl. The three R groups may be the same or different.
Representative examples of "trialkylsily1" include, but are not limited to,
trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl
(TBS
or TBDMS), and triisopropylsilyl (TIPS).
Asymmetric centers exist in the compounds of the present disclosure. These
centers are designated by the symbols "R" or "S", depending on the
configuration of
substituents around the chiral carbon atom. It should be understood that the
disclosure encompasses all stereochemical isomeric forms, or mixtures thereof,
which possess the ability to inhibit NS5A. Individual stereoisomers of
compounds
can be prepared synthetically from commercially available starting materials
which
contain chiral centers or by preparation of mixtures of enantiomeric products
followed by separation such as conversion to a mixture of diastereomers
followed by
separation or recrystallization, chromatographic techniques, or direct
separation of
enantiomers on chiral chromatographic columns. Starting compounds of
particular
stereochemistry are either commercially available or can be made and resolved
by
techniques known in the art.
Certain compounds of the present disclosure may also exist in different stable
conformational forms which may be separable. Torsional asymmetry due to
restricted rotation about an asymmetric single bond, for example because of
steric
hindrance or ring strain, may permit separation of different conformers. The
present
disclosure includes each conformational isomer of these compounds and mixtures
thereof
The term "compounds of the present disclosure", and equivalent expressions,
are meant to embrace compounds of Formula (I), and pharmaceutically acceptable
enantiomers, diastereomers, and salts thereof Similarly, references to
intermediates
are meant to embrace their salts where the context so permits.
The present disclosure is intended to include all isotopes of atoms occurring
in the present compounds. Isotopes include those atoms having the same atomic
number but different mass numbers. By way of general example and without
limitation, isotopes of hydrogen include deuterium and tritium. Isotopes of
carbon
include 13C and 14C. Isotopically-labeled compounds of the invention can
generally
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be prepared by conventional techniques known to those skilled in the art or by
processes analogous to those described herein, using an appropriate
isotopically-
labeled reagent in place of the non-labeled reagent otherwise employed. Such
compounds may have a variety of potential uses, for example as standards and
reagents in determining biological activity. In the case of stable isotopes,
such
compounds may have the potential to favorably modify biological,
pharmacological,
or pharmacokinetic properties.
The compounds of the present disclosure can exist as pharmaceutically
acceptable salts. The term "pharmaceutically acceptable salt," as used herein,
represents salts or zwitterionic forms of the compounds of the present
disclosure
which are water or oil-soluble or dispersible, which are, within the scope of
sound
medical judgment, suitable for use in contact with the tissues of patients
without
excessive toxicity, irritation, allergic response, or other problem or
complication
commensurate with a reasonable benefit/risk ratio, and are effective for their
intended
use. The salts can be prepared during the final isolation and purification of
the
compounds or separately by reacting a suitable nitrogen atom with a suitable
acid.
Representative acid addition salts include acetate, adipate, alginate,
citrate, aspartate,
benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate;
digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate,
fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate,
lactate, maleate, mesitylenesulfonate, methanesulfonate, naphthylenesulfonate,
nicotinate, 2-naphthalenesulfonate, oxalate, palmoate, pectinate, persulfate,
3-
phenylproprionate, picrate, pivalate, propionate, succinate, tartrate,
trichloroacetate,
trifluoroacetate, phosphate, glutamate, bicarbonate, para-toluenesulfonate,
and
undecanoate. Examples of acids which can be employed to form pharmaceutically
acceptable addition salts include inorganic acids such as hydrochloric,
hydrobromic,
sulfuric, and phosphoric, and organic acids such as oxalic, maleic, succinic,
and
citric.
Basic addition salts can be prepared during the final isolation and
purification
of the compounds by reacting a carboxy group with a suitable base such as the
hydroxide, carbonate, or bicarbonate of a metal cation or with ammonia or an
organic
primary, secondary, or tertiary amine. The cations of pharmaceutically
acceptable
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salts include lithium, sodium, potassium, calcium, magnesium, and aluminum, as
well as nontoxic quaternary amine cations such as ammonium,
tetramethylammonium, tetraethylammonium, methylamine, dimethylamine,
trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine,
pyridine,
N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine,
dicyclohexylamine,
procaine, dibenzylamine, N,N-dibenzylphenethylamine, and N,N'-
dibenzylethylenediamine. Other representative organic amines useful for the
formation of base addition salts include ethylenediamine, ethanolamine,
diethanolamine, piperidine, and piperazine.
When it is possible that, for use in therapy, therapeutically effective
amounts
of a compound of Formula (I), as well as pharmaceutically acceptable salts
thereof,
may be administered as the raw chemical, it is possible to present the active
ingredient as a pharmaceutical composition. Accordingly, the disclosure
further
provides pharmaceutical compositions, which include therapeutically effective
amounts of compounds of Formula (I) or pharmaceutically acceptable salts
thereof,
and one or more, preferably one to three, pharmaceutically acceptable
carriers,
diluents, or excipients. The term "therapeutically effective amount," as used
herein,
refers to the total amount of each active component that is sufficient to show
a
meaningful patient benefit, e.g., a sustained reduction in viral load. When
applied to
an individual active ingredient, administered alone, the term refers to that
ingredient
alone. When applied to a combination, the term refers to combined amounts of
the
active ingredients that result in the therapeutic effect, whether administered
in
combination, serially, or simultaneously. The compounds of Formula (I) and
pharmaceutically acceptable salts thereof, are as described above. The
carrier(s),
diluent(s), or excipient(s) must be acceptable in the sense of being
compatible with
the other ingredients of the formulation and not deleterious to the recipient
thereof
In accordance with another aspect of the present disclosure there is also
provided a
process for the preparation of a pharmaceutical formulation including admixing
a
compound of Formula (I), or a pharmaceutically acceptable salt thereof, with
one or
more, preferably one to three, pharmaceutically acceptable carriers, diluents,
or
excipients. The term "pharmaceutically acceptable," as used herein, refers to
those
compounds, materials, compositions, and/or dosage forms which are, within the
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scope of sound medical judgment, suitable for use in contact with the tissues
of
patients without excessive toxicity, irritation, allergic response, or other
problem or
complication commensurate with a reasonable benefit/risk ratio, and are
effective for
their intended use.
Pharmaceutical formulations may be presented in unit dose forms containing
a predetermined amount of active ingredient per unit dose. Dosage levels of
between
about 0.01 and about 250 milligram per kilogram ("mg/kg") body weight per day,
preferably between about 0.05 and about 100 mg/kg body weight per day of the
compounds of the present disclosure are typical in a monotherapy for the
prevention
and treatment of HCV mediated disease. Typically, the pharmaceutical
compositions
of this disclosure will be administered from about 1 to about 5 times per day
or
alternatively, as a continuous infusion. Such administration can be used as a
chronic
or acute therapy. The amount of active ingredient that may be combined with
the
carrier materials to produce a single dosage form will vary depending on the
condition being treated, the severity of the condition, the time of
administration, the
route of administration, the rate of excretion of the compound employed, the
duration
of treatment, and the age, gender, weight, and condition of the patient.
Preferred unit
dosage formulations are those containing a daily dose or sub-dose, as herein
above
recited, or an appropriate fraction thereof, of an active ingredient.
Generally,
treatment is initiated with small dosages substantially less than the optimum
dose of
the compound. Thereafter, the dosage is increased by small increments until
the
optimum effect under the circumstances is reached. In general, the compound is
most desirably administered at a concentration level that will generally
afford
antivirally effective results without causing any harmful or deleterious side
effects.
When the compositions of this disclosure comprise a combination of a
compound of the present disclosure and one or more additional therapeutic or
prophylactic agent, both the compound and the additional agent are usually
present at
dosage levels of between about 10 to 150%, and more preferably between about
10
and 80% of the dosage normally administered in a monotherapy regimen.
Pharmaceutical formulations may be adapted for administration by any
appropriate route, for example by the oral (including buccal or sublingual),
rectal,
nasal, topical (including buccal, sublingual, or transdermal), vaginal, or
parenteral
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(including subcutaneous, intracutaneous, intramuscular, intra-articular,
intrasynoyial,
intrasternal, intrathecal, intralesional, intravenous, or intradermal
injections or
infusions) route. Such formulations may be prepared by any method known in the
art
of pharmacy, for example by bringing into association the active ingredient
with the
carrier(s) or excipient(s). Oral administration or administration by injection
are
preferred.
Pharmaceutical formulations adapted for oral administration may be
presented as discrete units such as capsules or tablets; powders or granules;
solutions
or suspensions in aqueous or non-aqueous liquids; edible foams or whips; or
oil-in-
water liquid emulsions or water-in-oil emulsions.
For instance, for oral administration in the form of a tablet or capsule, the
active drug component can be combined with an oral, non-toxic pharmaceutically
acceptable inert carrier such as ethanol, glycerol, water, and the like.
Powders are
prepared by comminuting the compound to a suitable fine size and mixing with a
similarly comminuted pharmaceutical carrier such as an edible carbohydrate,
as, for
example, starch or mannitol. Flavoring, preservative, dispersing, and coloring
agent
can also be present.
Capsules are made by preparing a powder mixture, as described above, and
filling formed gelatin sheaths. Glidants and lubricants such as colloidal
silica, talc,
magnesium stearate, calcium stearate, or solid polyethylene glycol can be
added to
the powder mixture before the filling operation. A disintegrating or
solubilizing
agent such as agar-agar, calcium carbonate, or sodium carbonate can also be
added to
improve the availability of the medicament when the capsule is ingested.
Moreover, when desired or necessary, suitable binders, lubricants,
disintegrating agents, and coloring agents can also be incorporated into the
mixture.
Suitable binders include starch, gelatin, natural sugars such as glucose or
beta-
lactose, corn sweeteners, natural and synthetic gums such as acacia,
tragacanth or
sodium alginate, carboxymethylcellulose, polyethylene glycol, and the like.
Lubricants used in these dosage forms include sodium oleate, sodium chloride,
and
the like. Disintegrators include, without limitation, starch, methyl
cellulose, agar,
betonite, xanthan gum, and the like. Tablets are formulated, for example, by
preparing a powder mixture, granulating or slugging, adding a lubricant and
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disintegrant, and pressing into tablets. A powder mixture is prepared by
mixing the
compound, suitable comminuted, with a diluent or base as described above, and
optionally, with a binder such as carboxymethylcellulose, an aliginate,
gelating, or
polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption
accelerator
such as a quaternary salt and/or and absorption agent such as betonite,
kaolin, or
dicalcium phosphate. The powder mixture can be granulated by wetting with a
binder such as syrup, starch paste, acadia mucilage, or solutions of
cellulosic or
polymeric materials and forcing through a screen. As an alternative to
granulating,
the powder mixture can be run through the tablet machine and the result is
imperfectly formed slugs broken into granules. The granules can be lubricated
to
prevent sticking to the tablet forming dies by means of the addition of
stearic acid, a
stearate salt, talc, or mineral oil. The lubricated mixture is then compressed
into
tablets. The compounds of the present disclosure can also be combined with a
free
flowing inert carrier and compressed into tablets directly without going
through the
granulating or slugging steps. A clear or opaque protective coating consisting
of a
sealing coat of shellac, a coating of sugar or polymeric material, and a
polish coating
of wax can be provided. Dyestuffs can be added to these coatings to
distinguish
different unit dosages.
Oral fluids such as solution, syrups, and elixirs can be prepared in dosage
unit
form so that a given quantity contains a predetermined amount of the compound.
Syrups can be prepared by dissolving the compound in a suitably flavored
aqueous
solution, while elixirs are prepared through the use of a non-toxic vehicle.
Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and
polyoxyethylene sorbitol ethers, preservatives, flavor additive such as
peppermint oil
or natural sweeteners, or saccharin or other artificial sweeteners, and the
like can also
be added.
Where appropriate, dosage unit formulations for oral administration can be
microencapsulated. The formulation can also be prepared to prolong or sustain
the
release as for example by coating or embedding particulate material in
polymers,
wax, or the like.
The compounds of Formula (I), and pharmaceutically acceptable salts thereof,
can also be administered in the form of liposome delivery systems, such as
small
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unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles.
Liposomes can be formed from a variety of phopholipids, such as cholesterol,
stearylamine, or phophatidylcholines.
The compounds of Formula (I) and pharmaceutically acceptable salts thereof
may also be delivered by the use of monoclonal antibodies as individual
carriers to
which the compound molecules are coupled. The compounds may also be coupled
with soluble polymers as targetable drug carriers. Such polymers can include
polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidephenol,
polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine substituted
with palitoyl residues. Furthermore, the compounds may be coupled to a class
of
biodegradable polymers useful in achieving controlled release of a drug, for
example,
polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid,
polyorthoesters,
polyacetals, polydihydropyrans, polycyanoacrylates, and cross-linked or
amphipathic
block copolymers of hydrogels.
Pharmaceutical formulations adapted for transdermal administration may be
presented as discrete patches intended to remain in intimate contact with the
epidermis of the recipient for a prolonged period of time. For example, the
active
ingredient may be delivered from the patch by iontophoresis as generally
described in
Pharm. Res., 3(6):318 (1986).
Pharmaceutical formulations adapted for topical administration may be
formulated as ointments, creams, suspensions, lotions, powders, solutions,
pastes,
gels, sprays, aerosols, or oils.
For treatments of the eye or other external tissues, for example mouth and
skin, the formulations are preferably applied as a topical ointment or cream.
When
formulated in an ointment, the active ingredient may be employed with either a
paraffinic or a water-miscible ointment base. Alternatively, the active
ingredient
may be formulated in a cream with an oil-in-water cream base or a water-in oil
base.
Pharmaceutical formulations adapted for topical administrations to the eye
include
eye drops wherein the active ingredient is dissolved or suspended in a
suitable
carrier, especially an aqueous solvent.
Pharmaceutical formulations adapted for topical administration in the mouth
include lozenges, pastilles, and mouth washes.
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Pharmaceutical formulations adapted for rectal administration may be
presented as suppositories or as enemas.
Pharmaceutical formulations adapted for nasal administration wherein the
carrier is a solid include a course powder having a particle size for example
in the
range 20 to 500 microns which is administered in the manner in which snuff is
taken,
i.e., by rapid inhalation through the nasal passage from a container of the
powder
held close up to the nose. Suitable formulations wherein the carrier is a
liquid, for
administration as a nasal spray or nasal drops, include aqueous or oil
solutions of the
active ingredient.
Pharmaceutical formulations adapted for administration by inhalation include
fine particle dusts or mists, which may be generated by means of various types
of
metered, dose pressurized aerosols, nebulizers, or insufflators.
Pharmaceutical formulations adapted for vaginal administration may be
presented as pessaries, tampons, creams, gels, pastes, foams, or spray
formulations.
Pharmaceutical formulations adapted for parenteral administration include
aqueous and non-aqueous sterile injection solutions which may contain anti-
oxidants,
buffers, bacteriostats, and soutes which render the formulation isotonic with
the
blood of the intended recipient; and aqueous and non-aqueous sterile
suspensions
which may include suspending agents and thickening agents. The formulations
may
be presented in unit-dose or multi-dose containers, for example sealed
ampoules and
vials, and may be stored in a freeze-dried (lyophilized) condition requiring
only the
addition of the sterile liquid carrier, for example water for injections,
immediately
prior to use. Extemporaneous injection solutions and suspensions may be
prepared
from sterile powders, granules, and tablets.
It should be understood that in addition to the ingredients particularly
mentioned above, the formulations may include other agents conventional in the
art
having regard to the type of formulation in question, for example those
suitable for
oral administration may include flavoring agents.
The term "patient" includes both human and other mammals.
The term "treating" refers to: (i) preventing a disease, disorder or condition
from occurring in a patient that may be predisposed to the disease, disorder,
and/or
condition but has not yet been diagnosed as having it; (ii) inhibiting the
disease,
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disorder, or condition, i.e., arresting its development; and (iii) relieving
the disease,
disorder, or condition, i.e., causing regression of the disease, disorder,
and/or
condition.
The compounds of the present disclosure can also be administered with a
cyclosporin, for example, cyclosporin A. Cyclosporin A has been shown to be
active
against HCV in clinical trials (Hepatology, 38:1282 (2003); Biochem. Biophys.
Res.
Commun., 313:42 (2004); J. Gastroenterol., 38:567 (2003)).
Table 1 below lists some illustrative examples of compounds that can be
administered with the compounds of this disclosure. The compounds of the
disclosure can be administered with other anti-HCV activity compounds in
combination therapy, either jointly or separately, or by combining the
compounds
into a composition.
Table 1
Brand Name Physiological Class Type of Inhibitor or Source
Company
Target
NIM811 Cyclophilin inhibitors Novartis
Debiopharm
Debio-025
Zadaxin Immunomodulator SciClone
Suvus Methylene blue Bioenvision
Actilon (CPG10101) TLR9 agonist Coley
Batabulin (T67) Anticancer f3-Tubulin inhibitor Tularik Inc.,
South
San Francisco, CA
ISIS 14803 Antiviral Antisense ISIS Pharmaceuticals
Inc, Carlsbad, CA /
Elan Pharmaceuticals
Inc., New York, NY
Summetrel Antiviral Antiviral Endo Pharmaceuticals
Holdings Inc., Chadds
Ford, PA
GS-9132 (ACH-806) Antiviral HCV inhibitor Achillion / Gilead
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Brand Name Physiological Class Type of Inhibitor or Source
Company
Target
Pyrazolopyrimidine Antiviral HCV inhibitors Arrow Therapeutics
compounds and salts Ltd.
From
WO 2005/047288
May 26, 2005
Levovirin Antiviral IMPDH inhibitor Ribapharm Inc., Costa
Mesa, CA
Merimepodib Antiviral IMPDH inhibitor Vertex
(VX-497) Pharmaceuticals Inc,
Cambridge, MA
XTL-6865 Antiviral Monoclonal antibody XTL
(XTL-002) Biopharmaceuticals
Ltd., Rehovot, Israel
Telaprevir Antiviral N53 serine protease Vertex
(VX-950, inhibitor Pharmaceuticals Inc,
LY-570310) Cambridge, MA / Eli
Lilly and Co., Inc.,
Indianapolis, IN
HCV-796 Antiviral NS5B replicase Wyeth / Viropharma
inhibitor
NM-283 Antiviral NS5B replicase Idenix / Novartis
inhibitor
GL-59728 Antiviral NS5B replicase Gene Labs / Novartis
inhibitor
GL-60667 Antiviral NS5B replicase Gene Labs / Novartis
inhibitor
2'C MeA Antiviral NS5B replicase Gilead
inhibitor
PSI 6130 Antiviral NS5B replicase Roche
inhibitor
R1626 Antiviral NS5B replicase Roche
inhibitor
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Brand Name Physiological Class Type of Inhibitor or Source
Company
Target
2'C Methyl Antiviral NS5B replicase Merck
adenosine inhibitor
JTK-003 Antiviral RdRp inhibitor Japan Tobacco Inc.,
Tokyo, Japan
Leyovirin Antiviral Ribavirin ICN Pharmaceuticals,
Costa Mesa, CA
Ribavirin Antiviral Ribavirin Schering-Plough
Corporation,
Kenilworth, NJ
Viramidine Antiviral Ribavirin prodrug Ribapharm Inc.,
Costa
Mesa, CA
Heptazyme Antiviral Ribozyme Ribozyme
Pharmaceuticals Inc.,
Boulder, CO
BILN-2061 Antiviral Serine protease Boehringer Ingelheim
inhibitor Pharma KG,
Ingelheim, Germany
SCH 503034 Antiviral Serine protease Schering-Plough
inhibitor
Zadazim Immune modulator Immune modulator SciClone
Pharmaceuticals Inc.,
San Mateo, CA
Ceplene Immunomodulator Immune modulator Maxim
Pharmaceuticals Inc.,
San Diego, CA
CELLCEPTO Immunosuppressant HCV IgG F. Hoffmann-La
immunosuppressant Roche LTD, Basel,
Switzerland
Ciyacir Immunosuppressant HCV IgG Nabi
immunosuppressant Biopharmaceuticals
Inc., Boca Raton, FL
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Brand Name Physiological Class Type of Inhibitor or Source
Company
Target
Albuferon - a Interferon Albumin IFN-a2b Human Genome
Sciences Inc.,
Rockville, MD
Infergen A Interferon IFN alfacon-1 InterMune
Pharmaceuticals Inc.,
Brisbane, CA
Omega IFN Interferon IFN-o) Intarcia Therapeutics
IFN-f3 and EMZ701 Interferon IFN-f3 and EMZ701 Transition
Therapeutics Inc.,
Ontario, Canada
REBIFO Interferon IFN- '31 a Serono, Geneva,
Switzerland
Roferon A Interferon IFN-a2a F. Hoffmann-La
Roche LTD, Basel,
Switzerland
Intron A Interferon IFN-a2b Schering-Plough
Corporation,
Kenilworth, NJ
Intron A and Zadaxin Interferon IFN-a2b/a1-thymosin RegeneRx Biopharma.
Inc., Bethesda, MD /
SciClone
Pharmaceuticals Inc,
San Mateo, CA
Rebetron Interferon IFN-a2b / ribavirin Schering-Plough
Corporation,
Kenilworth, NJ
Actimmune Interferon INF-y InterMune Inc.,
Brisbane, CA
Interferon-f3 Interferon Interferon-f3-1a Serono
Multiferon Interferon Long lasting IFN Viragen / Valentis
WelHeron Interferon Lymphoblastoid IFN- GlaxoSmithKline plc,
anl Uxbridge, UK
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Brand Name Physiological Class Type of Inhibitor or Source
Company
Target
Omniferon Interferon natural IFN-a Viragen Inc.,
Plantation, FL
Pegasys Interferon PEGylated IFN-a2a F. Hoffmann-La
Roche LTD, Basel,
Switzerland
Pegasys and Ceplene Interferon PEGylated IFN- Maxim
a2a/immune Pharmaceuticals Inc.,
modulator San Diego, CA
Pegasys and Interferon PEGylated IFN- F. Hoffmann-La
Ribavirin a2a/ribavirin Roche LTD, Basel,
Switzerland
PEG-Intron Interferon PEGylated IFN-a2b Schering-Plough
Corporation,
Kenilworth, NJ
PEG-Intron / Interferon PEGylated IFN- Schering-Plough
Ribavirin a2b/ribavirin Corporation,
Kenilworth, NJ
IP-501 Liver protection Antifibrotic Indeyus
Pharmaceuticals Inc.,
Lexington, MA
IDN-6556 Liver protection Caspase inhibitor Idun
Pharmaceuticals
Inc., San Diego, CA
ITMN-191 Antiviral Serine protease InterMune
(R-7227) inhibitor Pharmaceuticals Inc.,
Brisbane, CA
GL-59728 Antiviral NS5B replicase Genelabs
inhibitor
ANA-971 Antiviral TLR-7 agonist Anadys
Boceprevir Antiviral Serine protease Schering-Plough
inhibitor
TMS-435 Antiviral Serine protease Tibotec BVBA,
inhibitor Mechelen, Belgium
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Brand Name Physiological Class Type of Inhibitor or Source
Company
Target
BI-201335 Antiviral Serine protease Boelu-inger Ingelheim
inhibitor Pharma KG,
Ingelheim, Germany
MK-7009 Antiviral Serine protease Merck
inhibitor
PF-00868554 Antiviral Replicase inhibitor Pfizer
ANA598 Antiviral Non-Nucleoside Anadys
NS5B polymerase Pharmaceuticals, Inc.,
inhibitor San Diego, CA, USA
IDX375 Antiviral Non-Nucleoside Idenix
replicase inhibitor Pharmaceuticals,
Cambridge, MA, USA
BILB 1941 Antiviral NS5B polymerase Boelu-inger Ingelheim
inhibitor Canada Ltd R&D,
Laval, QC, Canada
PSI-7851 Antiviral Nucleoside Pharmasset,
polymerase inhibitor Princeton, NJ, USA
VCH-759 Antiviral NS5B polymerase ViroChem Pharma
inhibitor
VCH-916 Antiviral NS5B polymerase ViroChem Pharma
inhibitor
GS-9190 Antiviral NS5B polymerase Gilead
inhibitor
Peg-interferon lamda Antiviral Interferon ZymoGenetics /
Bristol-Myers Squibb
The compounds of the present disclosure may also be used as laboratory
reagents. Compounds may be instrumental in providing research tools for
designing
of viral replication assays, validation of animal assay systems and structural
biology
studies to further enhance knowledge of the HCV disease mechanisms. Further,
the
compounds of the present disclosure are useful in establishing or determining
the
binding site of other antiviral compounds, for example, by competitive
inhibition.
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The compounds of this disclosure may also be used to treat or prevent viral
contamination of materials and therefore reduce the risk of viral infection of
laboratory or medical personnel or patients who come in contact with such
materials,
e.g., blood, tissue, surgical instruments and garments, laboratory instruments
and
garments, and blood collection or transfusion apparatuses and materials.
This disclosure is intended to encompass compounds having Formula (I)
when prepared by synthetic processes or by metabolic processes including those
occurring in the human or animal body (in vivo) or processes occurring in
vitro.
The abbreviations used in the present application, including particularly in
the
illustrative examples which follow, are well-known to those skilled in the
art. Some
of the abbreviations used are as follows: TFA for trifluoroacetic acid; min or
min. or
mins for minutes; MeCN or ACN for acetonitrile; LDA for lithium
diisopropylamide;
DMSO for dimethylsulfoxide; h or hr or hrs for hours; Boc or BOC for tert-
butoxycarbonyl; HATU for 0-(7-azabenzotriazol-1-y1)-N,N,N',N'-
tetramethyluronium hexafluorophosphate; RT or Rt or rt for retention time or
room
temperature (context will dictate); Me for methyl; DMF for N,N-
dimethylformamide;
Pd(Ph3P)4 for tetrakistriphenyl phosphine palladium; Me0H for methanol; Me0D
for
CD40D; TEA for triethylamine; Ph for phenyl; TBDPS for tert-
butyldiphenylsilyl;
Et3N or TEA for triethylamine; DMAP for N,N-dimethylaminopyridine; Et0Ac for
ethyl acetate; TBAF for tetrabutylammonium fluoride; THF for tetrahydrofuran;
DIEA or DIPEA or iPr2NEt for diisopropylethylamine; NCS for N-
chlorosuccinimide; NBS for N-bromosuccinimide; DCM for dichloromethane; SEM
for 2-(trimethylsilyl)ethoxymethyl; DCE for 1,2-dichloroethane; EDCI for 1-
ethy1-3-
(3-dimethylaminopropyl)carbodiimide; DBU for 1,8-diazabicyclo[5.4.0]undec-7-
ene;
Pd(t-Bu3P)2 for palladium bis(tributylphosphine); HMDS for
hexamethyldisilazide;
TMSCHN2 for trimethylsilyldiazomethane; H-D-Ser-OBz1 for D-serine benzyl
ester;
i-PrOH for isopropanol; LiHMDS for lithium hexamethyldisilazide; DIBAL or
DIBALH for diisobutylaluminum hydride; TBDMS for tert-butyldimethylsilyl; CBz
for carbobenzyloxy; Bn for benzyl; DEAD for diethyl azodicarboxylate; mCPBA
for
meta-chloroperoxybenzoic acid; TMSCN for trimethylsilyl cyanide; dpppe for 1,5-
Bis(diphenylphosphino) pentane; TMEDA for tetramethylethylenediamine; OAc for
acetate; DMA for N,N-dimethylacetamide; and d for days.
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The present disclosure will now be described in connection with certain
embodiments which are not intended to limit its scope. On the contrary, the
present
disclosure covers all alternatives, modifications, and equivalents as can be
included
within the scope of the claims. Thus, the following examples, which include
specific
embodiments, will illustrate one practice of the present disclosure, it being
understood that the examples are for the purposes of illustration of certain
embodiments and are presented to provide what is believed to be the most
useful and
readily understood description of its procedures and conceptual aspects.
Starting materials can be obtained from commercial sources or prepared by well-
established literature methods known to those of ordinary skill in the art.
EXAMPLES
Unless noted otherwise, purity assessments were conducted on Shimadzu LC
system, and retention time (Rt) determination and low resolution mass analysis
were
conducted on a Shimadzu LC system coupled with Waters MICROMASSO ZQ MS
system. It should be noted that retention times may vary slightly between
machines.
Condition 1
Column = PHENOMENEXO, C18, 3.0 X 50 mm, 10 lam
Start %B = 0
Final %B = 100
Gradient time = 2 min
Stop time = 3 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1 % TFA in 10% methanol/90% water
Solvent B = 0.1 % TFA in 90% methanol/10% water
Condition 2
Column = XTERRAO, C18, 3.0 x 50 mm, S7
Start %B = 0
Final %B = 100
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Gradient time = 2 min
Stop time = 3 min
Flow Rate =5 mL/min
Wavelength = 220 nm
Solvent A = 0.2 % H3PO4 in 10% methanol/90% water
Solvent B = 0.2 % H3PO4 in 90% methanol/10% water
Condition 3
Column = PHENOMENEXO, C18, 3.0 X 50 mm, 10 p.m
Start %B = 0
Final %B = 100
Gradient time = 3 min
Stop time = 4 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1 % TFA in 10% methanol/90% water
Solvent B = 0.1 % TFA in 90% methanol/10% water
Condition 4
Column = XTERRAO, C18, 3.0 x 50 mm, S7
Start %B =0
Final %B = 100
Gradient time = 10 min
Stop time = 11 min
Flow Rate = 5 mL/min
Wavelength = 220 nm
Solvent A = 0.2 % H3PO4 in 10% methanol/90% water
Solvent B = 0.2 % H3PO4 in 90% methanol/10% water
Condition 5
Column = PHENOMENEXO, C18, 3.0 X 50 mm, 10 p.m
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Start %B =0
Final %B = 100
Gradient time = 4 min
Stop time = 5 min
Flow Rate = 4 mL/min
Wavelength = 220 nm
Solvent A = 0.1 % TFA in 10% methanol/90% water
Solvent B = 0.1 % TFA in 90% methanol/10% water
Condition 6
Column = Phenomenex-Luna, C18, 4.6 X 50 mm, S10
Start %B =0
Final %B = 100
Gradient time = 3 min
Stop time = 4 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1 % TFA in 10% methanol/90% water
Solvent B = 0.1 % TFA in 90% methanol/10% water
Condition 7
Column = Phenomenex-Luna, C18, 3.0X 50 mm, S10
Start %B =0
Final %B = 100
Gradient time = 2 min
Stop time = 3 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1 % TFA in 10% methanol/90% water
Solvent B = 0.1 % TFA in 90% methanol/10% water
Condition 9
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Column = Waters Sunfire, C18, 4.6X150 mm, 3.5 [im
Start %B= 10
Final %B = 50
Gradient time = 20 min
Stop time = 25 min
Flow Rate = 1 mL/min
Wavelength = 220 & 254 nm
Slovent A = 0.1% TFA in 5% CH3CN/95%H20
Solvent B = 0.1% TFA in 95% CH3CN/5% H20
Condition 9a
Same as Condition 9 with the exception that Stop time = 35 min
Condition 9a.1
Column = Waters Sunfire, C18, 4.6X150 mm, 3.5 [im
Start %B = 30
Final %B = 100
Gradient time = 20 min
Stop time = 25 min
Flow Rate = 1 mL/min
Wavelength = 220 & 254 nm
Slovent A = 0.1% TFA in 5% CH3CN/95%H20
Solvent B = 0.1% TFA in 95% CH3CN/5% H20
Condition 10
Column = Waters Xbridge phenyl, C18, 4.6X150 mm, 3 [im
Start %B= 10
Final %B = 50
Gradient time = 20 min
Stop time = 25 min
Flow Rate = 1 mL/min
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Wavelength = 220 & 254 nm
Slovent A = 0.1% TFA in 5% CH3CN/95%H20
Solvent B = 0.1% TFA in 95% CH3CN/5% H20
Condition 10a
Same as Condition 10 with the exception that Stop time = 35 min
Condition 10a.1
Column = Waters Xbridge phenyl, C18, 4.6X150 mm, 3 p.m
Start %B = 40
Final %B = 100
Gradient time = 20 min
Stop time = 25 min
Flow Rate = 1 mL/min
Wavelength = 220 & 254 nm
Slovent A = 0.1% TFA in 5% CH3CN/95%H20
Solvent B = 0.1% TFA in 95% CH3CN/5% H20
Condition 10b
Column = Sunfire, C18, 3.0 X 150 mm, 3.5 p.m
Start %B= 10
Final %B = 40
Gradient time = 15 min
Stop time = 18 min
Flow Rate = 1 mL/min
Wavelength 1 = 220 nm
Wavelength 2 = 254 nm
Solvent A = 0.1 % TFA in 5% MeCN/95% water
Solvent B = 0.1 % TFA in 95% MeCN/5% water
Condition 10c
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Column = Xbridge Phenyl, C18, 3.0 X 150 mm, 3.5 pm
Start %B= 10
Final %B = 40
Gradient time = 15 min
Stop time = 18 min
Flow Rate = 1 mL/min
Wavelength 1 = 220 nm
Wavelength 2 = 254 nm
Solvent A = 0.1 % TFA in 5% MeCN/95% water
Solvent B = 0.1 % TFA in 95% MeCN/5% water
Condition 10d
Column = PHENOMENEXO-Luna, C18, 2.0 X 50 mm, 3 pm
Start %B =0
Final %B = 100
Gradient time = 4 min
Stop time = 5 min
Flow Rate = 0.8 mL/min
Wavelength = 220 nm
Solvent A = 0.1 % TFA in 10% methanol/90% water
Solvent B = 0.1 % TFA in 90% methanol/10% water
Oven temp. = 40 C
Condition 10e (Agilent 1200 series LC system)
Column = Xbridge phenyl, C18, 4.6X150 mm, 3.5 pm
Solvent A = Buffer: CH3CN (95:5)
Solvent B = Buffer: CH3CN (5:95)
Buffer = 0.05% TFA in H20 (pH 2.5, adjusted with dilute ammonia)
Start %B= 10
Final %B = 100
Gradient time = 12 min
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Isocratic time = 3 min
Stop time = 23 min
Flow Rate = 1 mL/min
Wavelength = 220 & 254 nm
Condition 10e.1 (for a homogeneity index assessment on a Agilent 1200 series
LC
system)
Column = Sunfire C18 ((4.6x150) mm, 3.5 p.m)
Slovent A = Buffer: CH3CN (95:5)
Solvent B = Buffer: CH3CN (5:95)
Buffer = 0.05% TFA in H20 (pH 2.5, adjusted with dilute ammonia)
Start %B =10
Final %B = 100
Gradient time = 12 min
Isocratic time = 3 min
Stop time = 23 min
Flow Rate = 1 mL/min
Wavelength = 220 & 254 nm
Condition 10e.2 (for a homogeneity index assessment on a Agilent 1200 series
LC
system)
Column = Sunfire C18 ((4.6x150) mm, 3.5 p.m)
Slovent A = Buffer: CH3CN (95:5)
Solvent B = Buffer: CH3CN (5:95)
Buffer = 0.05% TFA in H20 (pH 2.5, adjusted with dilute ammonia)
Start %B =0
Final %B =50
Gradient time-1 = 15 min
Final %B = 100
Gradient time-2 = 3 min
Isocratic time = 5 min
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Stop time = 28 min
Flow Rate = 1 mL/min
Wavelength = 220 & 254 nm
Condition 10e.3 (for a homogeneity index assessment on a Agilent 1200 series
LC
system)
Column = Xbridge phenyl ((4.6x150) mm, 3.5 p.m)
Solvent A = 10 mM NH4HCO3 in H20 (pH 9.5, adjusted with dilute ammonia)
Solvent B = Me0H
Start %B =10
Final %B = 100
Gradient time = 12 min
Isocratic time = 8 min
Stop time = 23 min
Flow Rate = 1 mL/min
Wavelength = 220 & 254 nm
Condition 10e.4 (for a homogeneity index assessment on a Agilent 1200 series
LC
system)
Column = Xbridge phenyl ((4.6x150) mm, 3.5 p.m)
Solvent A = Buffer: CH3CN (95:5)
Solvent B = Buffer: CH3CN (5:95)
Buffer = 0.05% TFA in H20 (pH 2.5, adjusted with dilute ammonia)
Start %B =10
Final %B = 100
Gradient time = 25 min
Isocratic time = 5 min
Stop time = 36 min
Flow Rate = 1 mL/min
Wavelength = 220 & 254 nm
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Condition 10e.5 (LC-MS analysis on a Agilent LC-1200 series coupled with 6140
single quad. mass spectrometer, ESI +ve mode, MS range: 100-2000)
Column = Zorbax SB C18 ((4.6x50) mm, 5 pm)
Slovent A = Me0H (10%) + 0.1% TFA in H20 (90%)
Solvent B = Me0H (90%) + 0.1% TFA in H20 (10%)
Start %B =0
Final %B = 100
Gradient time = 2 min
Stop time = 3 min
Flow Rate = 5 mL/min
Wavelength = 220 nm
Condition 10e.6 (LC-MS analysis on a Agilent LC-1200 series coupled with 6140
single quad. mass spectrometer, ESI +ve mode & -ye mode, MS range: 100-2000)
Column = Purosphergstar RP-18 ((4.0x55) mm, 3 m)
Slovent A = ACN (10%) +20 mM NH40Ac in H20 (90%)
Solvent B = ACN (90%) +20 mM NH40Ac in H20 (10%)
Start %B =0
Final %B = 100
Gradient time = 2.0 min
Isocratic time = 0.5 min
Stop time = 3 min
Flow Rate = 2.5 mL/min
Wavelength = 220 nm
Condition 10e.7 (LC-MS analysis on a Agilent LC-1200 series coupled with 6330
ion trap mass spectrometer, ESI +ve mode & -ye mode, MS range: 100-2000)
Column = Ascentis Express C18 (2.1X50 mm, 2.7 m)
Slovent A = CH3CN (2%) + 10 mM NH4COOH in H20 (98%)
Solvent B = CH3CN (98%) + 10 mM NH4COOH in H20 (2%)
Start %B =0
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Final %B = 100
Gradient time = 1.5 min
Isocratic time = 1.7 min
Stop time = 4 min
Flow Rate = 1.0 mL/min
Wavelength = 220 nm
Condition 10f (Agilent LC-1200 series coupled with 6140 single quad. mass
spectrometer, ESI +ve mode)
Column = Zorbax SB, C18, 4.6X50 mm, 5 pm
Slovent A = Me0H (10%) + 0.1% TFA in H20 (90%)
Solvent B = Me0H (90%) + 0.1% TFA in H20 (10%)
Start %B =0
Final %B = 100
Gradient time = 3 min
Isocratic time = 1 min
Stop time = 5 min
Flow Rate =5 mL/min
Wavelength = 220 nm
Condition lOg (Agilent LC-1200 series coupled with 6140 single quad. mass
spectrometer, ESI +ve mode)
Column = Ascentis Express, C-8, 2.1X5 mm, 2.7 p.m
Slovent A = CH3CN (2%) + 10 mM NH4COOH in H20 (98%)
Solvent B = CH3CN (98%) + 10 mM NH4COOH in H20 (2%)
Start %B =0
Final %B = 100
Gradient time = 1.5 min
Isocratic time = 1.7 min
Stop time = 4 min
Flow Rate = 1 mL/min
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Wavelength = 220 nm
Condition 10h
Column = PHENOMENEXO-Luna, C18, 4.6 X 30 mm, S10
Start %B = 0
Final %B = 100
Gradient time = 3 min
Stop time = 4 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1 % TFA in 10% methanol/90% water
Solvent B = 0.1 % TFA in 90% methanol/10% water
Condition 11
Column = PHENOMENEXO-Luna, C18, 50X2 mm, 3[im
Start %B =0
Final %B = 100
Gradient time = 4 min
Stop time = 5 min
Flow Rate = 0.8 mL/min
Wavelength = 220 nm
Solvent A = 5% methanol/95% water: 10 mM ammonium acetate
Solvent B = 95% methanol/5% water: 10 mM ammonium acetate
Oven temperature = 40 C
Condition 12 (Waters Acquity HPLC with Waters PDA UV-Vis detection and
Waters SQ MS-ESCI probe)
Column = Waters Acquity BEH, C18, 150 X 2.1 mm ID, 1.7 [im (at 35 C)
Mobile phase A = 0.05% TFA in water
Mobile phase B = 0.05% TFA in acetonitrile
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Solvent system: Hold 10%B: 0-1min; 10-98%B: 1-32min; Hold 98%B: 32-35 min;
98-10%B: 35-35.3 min; hold 10%B: 35.3-40 min
Flow rate = 0.35 ml/min
UV detection = 335 nm
EXAMPLES
Example OL-1
0 I* ri
/ 0 N \
--N )¨NH N
0
--N
Example OL-1, step a
0
CI CI
0 0
4,4'-Oxybis-(benzoic acid) dimethyl ester (1.5 g, 5.24 mmol) was added to a
solution of chloroiodomethane (3.05 mL, 41.92 mmol) in tetrahydrofuran (50 mL)
and the resulting solution was cooled to -78 C. A 1.8M solution of LDA (29
mL, 52
mmol) in tetrahydrofuran was added dropwise and the resulting brown suspension
was stirred at -78 C for 15 min. A solution of glacial acetic acid in
tetrahydrofuran
(10 mL in 50 mL) was then added slowly and the brown mixture was stirred at -
78 C
for 10 min. before allowing it to reach room temperature. The mixture was
taken up
in ethyl acetate and brine (1:1, 50 mL) and the organic layer was then
separated,
washed with a sat. solution of sodium bicarbonate and brine, dried (Mg504),
filtered,
and concentrated in vacuo. The residue was submitted to flash chromatography
(silica gel; 15 % ethyl acetate/hexanes) to provide OL-la as a pale brown
solid (0.6
g). 1H NMR (500 MHz, DMSO-d6) 6 ppm 5.17 (s, 4 H), 7.22 (d, J=8.85 Hz, 4 H),
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8.06 (d, J=8.85 Hz, 4 H). LC (Cond. 2): RT = 1.53 minutes, 97% homogeneity
index; LCMS: Anal. Calcd. For (M-H)+ C16H11C1203: 321.01; found: 321.30.
Example OL-1, step b
0 0 0
H2N NH2
0 0
2HCI
A mixture of OL-la (0.6 g, 1.85 mmol) and sodium diformylamide (0.42 g,
4.45 mmol) in acetonitrile (20 mL) was heated to reflux for 4 h. The solvent
was the
removed under reduced pressure and the remaining residue was redissolved in a
5%
HC1 solution in ethanol (30 mL) and heated to reflux temperature for 2 h. The
mixture was cooled in an ice-water bath and the resulting precipitate was
filtered,
washed with ethanol and ether and dried in vacuo. The recovered pale brow
solid
was used without further purification (0.66 g). 1H NMR (500 MHz, DMSO-d6) 6
ppm
4.54 (s, 4 H), 7.25 (d, J=8.85 Hz, 4 H), 8.11 (d, J=8.55 Hz, 4 H), 8.52 (br.
s, 6 H).
LC (Cond. 1): RT = 1.24 min; LRMS: Anal. Calcd. For (M+H)+ C16H17N203 285.12;
found: 285.19.
Example OL-1, step c
0 0 0
0 0
0)HN 0 H
. \
N---/
0
'Boo Boc'
N,N-Diisopropylethylamine (1.13 mL, 6.47 mmol) was added dropwise, over
15 minutes, to a heterogeneous mixture of N-Boc-L-proline (0.8 g, 3.73 mmol),
HATU (1.48 g, 3.88 mmol), OL-lb (0.66 g, 1.85 mmol) and dimethylformamide (15
mL), and stirred at ambient condition for 4 h. Most of the volatile components
were
removed in vacuo, and the resulting residue was partitioned between ethyl
acetate (30
mL) and water (20 mL). The organic layer was washed with water (20 mL) and
brine, dried (MgSO4), filtered, and concentrated in vacuo. A silica gel mesh
was
prepared from the residue and submitted to flash chromatography (silica gel;
65-85 %
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ethyl acetate/ hexanes) to provide OL-lc as a pale brown solid (0.7 g). 1H NMR
(500
MHz, DMSO-d6) 6 ppm 1.39/1.34 (rotomers, s, 18 H), 1.70 - 1.91 (m, 6 H), 2.00 -
2.25 (m, J=8.55 Hz, 2 H), 3.22 - 3.33 (m, 2 H), 3.32 - 3.47 (m, J=10.68, 10.68
Hz, 2
H), 4.08 -4.26 (m, 2 H), 4.56 (ddd, J=36.93, 18.16, 5.34 Hz, 4 H), 7.19 (d,
J=8.85
Hz, 4 H), 8.06 (d, J=7.93 Hz, 4 H), 8.11 -8.24 (m, J=5.49 Hz, 2 H). LC (Cond.
1):
RT = 2.55 min; LRMS: Anal. Calcd. for (M+H)+C36H47N409 679.33; found: 679.44.
Example OL-1, step d
N0 0 0
C)---_. 1 \N\>0
Ni HN NH N
i
Boc Boc
A mixture of OL-lc (0.7 g, 1.025 mmol) and ammonium acetate (0.79 g, 10
mmol) in xylenes (5 mL) was heated in a sealed tube at 140 C for 1.2 hours
under
microwave radiation. The volatile component was removed in vacuo, and the
residue
was partitioned carefully between ethyl acetate and water, whereby enough
saturated
sodium bicarbonate solution was added so as to make the pH of the aqueous
phase
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Example OL-1, step e
40 0 40
fl <N ---- i 1 N,C
N HN NH N
H 4 HCI H
To a solution of OL-ld (0.13 g, 0.2 mmol) in 30 mL dichloromethane was
added 1 mL of a 4.0M solution of HC1 in dioxane. The reaction was stirred for
2
hours at room temperature and concentrated under reduced pressure. The
resulting
residue was redissolved in a minimum amount of methanol and the desired
product
was triturated with ether, filtered and dried in vacuo. A pale tan solid
(0.11g) was
recovered and used without further purification. 1H NMR (500 MHz, DMSO-d6) 6
ppm 1.92 -2.07 (m, 2 H), 2.12 - 2.25 (m, 2 H), 2.37 -2.47 (m, 4 H), 3.30 -
3.47 (m, 4
Example OL-1
HATU (52 mg, 0.137 mmol) was added to a mixture of OL-le (35 mg, 0.06
mmol), diisopropylethylamine (58 L, 0.33 mmol) and Cap-1 (22 mg, 0.12 mmol)
in
dimethylformamide (3 mL), and the resulting mixture was stirred at ambient for
3 h.
The volatile component was removed in vacuo, and the residue was purified by a
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Examples OL-2 and OL-3
0
N
N\RA¨NH N y
0.\R
Examples OL-2 and OL-3 were prepared as TFA salts by substituting the
respective acids for Cap-1 using the same method described for Example OL-1.
Example 0 RT (LC-Cond.); MS data
R)LsS
Acid
OL-2 0 RT = 2.92 min. LC (Cond. 2); LCMS: Anal. Calcd. For
Phj=L
5S (M+H)+ C42H41N605 709.31; Found: 709.41, HRMS:
1-12-8
Anal. Calcd. for: C42H41N605 709.3 138; Found:
Mandelic acid 709.3147 (M+H)+
OL-3 0 RT = 3.65 min). LC (Cond. 2); LCMS: Anal. Calcd.
for
Ph j=L
5S (M+H) C46F147N807 823.35; Found: 823.66, HRMS:
Fli\Ty Anal. Calcd. for (M+H)+ C46H47N807 823.3568; Found:
0 823.3574
Cap-4
5
Example OL-4
0
\ 0 H
=
N
NN
\ 111\1 o
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Example OL-4, step a
0
I 0 0 0
0
I
0
0
Methyl 3-bromobenzoate (5 g, 23.2 mmol), methyl 4-hydroxybenzoate (5.3 g,
34.87 mmol), cesium carbonate (15.12 g, 46.4 mmol), copper (I) iodide (0.44 g,
2.32
mmol) and N,N-dimethyl hydrochloride (0.97 g, 6.96 mmol) were combined in
dioxane (100 ml) in a sealed tube and the resulting mixture was heated to 90
C for
h. The solvent was removed under reduced pressure and the residue was taken up
in ethyl acetate/water (1:1, 200 mL). The organic layer was washed with
saturated
aq. sodium carbonate, water and brine, dried (MgSO4), filtered and
concentrated in
10 vacuo. The resulting material was purified by flash chromatography
(silica gel, 5%
ethyl acetate/hexanes) to provide OL-4a as a clear oil that solidified upon
standing
(5.8 g). 1H NMR (500 MHz, DMSO-d6) 6 ppm 3.83 (s, 6 H), 7.10 (d, J=8.55 Hz, 2
H), 7.43 (dd, J=8.09, 2.59 Hz, 1 H), 7.56 (s, 1 H), 7.61 (t, J=7.93 Hz, 1 H),
7.81 (d,
J=7.63 Hz, 1 H), 7.98 (d, J=8.55 Hz, 2 H). LC (Cond. 2): RT = 1.53 minutes;
15 LCMS: Anal. Calcd. for (M+1)+ Ci6H1505: 287.01; found: 287.23.
Example OL-4, step b
0
0 0 0 CI
CI
0
OL-4b was prepared from OL-4a, according to the same procedure used for
the preparation of OL-la. 1H NMR (500 MHz, DMSO-d6) 6 ppm 5.14 (s, 2 H), 5.20
(s, 2 H), 7.13 (d, J=8.85 Hz, 2 H), 7.43 - 7.49 (m, 1 H), 7.64 (t, J=8.09 Hz,
1 H), 7.66
- 7.68 (m, 1 H), 7.85 (d, J=7.93 Hz, 1 H), 8.02 (d, J=8.85 Hz, 2 H). LC (Cond.
1):
RT = 1.51 min; LRMS: Anal. Calcd. for (M+H)+ C16H13C1203 323.02; found:
323.07.
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Example OL-4, step c
0
0 0 s NH2
H2N
0
2 HCI
OL-4c was prepared from Example OL-4b, according to the same procedure
used for the preparation of Example OL-lb. 1H NMR (500 MHz, DMSO-d6) 6 ppm.
4.52 (s, 2 H), 4.56 (s, 2 H), 7.16 (d, J=8.85 Hz, 2 H), 7.51 (dd, J=8.24, 2.44
Hz, 1 H),
7.68 (t, J=7.93 Hz, 1 H), 7.71 - 7.75 (m, 1 H), 7.91 (d, J=7.63 Hz, 1 H), 8.07
(d,
J=8.85 Hz, 2 H), 8.60 (s, 6 H). LC (Cond. 2): Anal. Calcd. for C16H17N203
(M+H)+
285.12; found: 285.15.
Example OL-4d
Boc,
0 Fl Nr1.1....)
0 410 0 0 N
0
N
CH 0
sBoc
Example OL-4d was prepared from Example OL-4c, according to the same
procedure used for the preparation of Example OL-lc. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.38/1.33 (rotomers, s, 18 H), 1.69 - 1.91 (m, 6 H), 1.95 -2.19 (m,
2 H),
3.20 - 3.31 (m, 2 H), 3.32 - 3.47 (m, 2 H), 4.08 - 4.23 (m, 2 H), 4.43 -4.68
(m, 4 H),
7.10 (d, J=8.55 Hz, 2 H), 7.42 (dd, J=8.09, 1.98 Hz, 1 H), 7.57 - 7.70 (m, 2
H), 7.86
(d, J=7.02 Hz, 1 H), 8.03 (d, J=7.93 Hz, 2 H), 8.08 - 8.24 (m, 2 H). LC (Cond.
1):
RT = 2.53 min; LRMS: Anal. Calcd. for C36H47N409 (M+H)+ 679.33; found: 679.42.
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Example OL-4e
Boc
NH N
I-----C
N 0 0 0
N
C)---- 1
NI HN
Boc
Example OL-4e was prepared from Example OL-4d, according to the same
procedure used for the preparation of Example OL-1d. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 1.14/1.17 /1.38/1.40(rotomers, s, 18 H), 1.71 - 2.06 (m, 6 H), 2.09 -
2.28
(m, J=25.79, 13.89 Hz, 2 H), 3.31 -3.40 (m, 2 H), 3.51 (s, 2 H), 4.65 -4.89
(m, 2 H),
6.80 (t, J=8.39 Hz, 1 H), 6.91 - 7.10 (m, 2 H), 7.28 - 7.55 (m, 5 H), 7.58 -
7.80 (m, 2
H), 11.66- 12.21 (m, 2 H). LC (Cond. 1): RT = 2.98 min; LRMS: Anal. Calcd. for
C36H45N605 (M+H)+ 641.34; found: 641.39.
Example OL-4f
NH HN
I ?----(3
N 0 0 0
N
C)---- i
N HN
H 4 HCI
Example OL-4f was prepared from Example OL-4e, according to the same
procedure used for the preparation of Example OL-le. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 1.94 - 2.06 (m, 2 H), 2.09 - 2.23 (m, 2 H), 2.28 - 2.38 (m, 1 H),
2.37 - 2.47
(m, 3 H), 3.27 - 3.50 (m, 4 H), 4.90 (br. s, 1 H), 4.99 (br. s, 1 H), 7.00 (d,
J=7.93 Hz,
1 H), 7.13 (d, J=8.55 Hz, 2 H), 7.49 (t, J=7.93 Hz, 1 H), 7.62 (s, 1 H), 7.69
(d, J=7.63
Hz, 1 H), 7.92 (d, J=8.55 Hz, 2 H), 7.99 (s, 2 H), 9.58 (br. s, 1 H), 9.84
(br. s, 1 H),
10.25 (br. d, J=22.28 Hz, 2 H). Note: the signal of the imidazole NH was too
broad to
assign a chemical shift. LC (Cond. 1): RT = 1.72 min; LRMS: Anal. Calcd. for
C26H29N60 (M+H)+ 441.24; found: 441.29.
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Example OL-4
Example OL-4 was prepared from Example OL-4f according to the same
procedure used for the preparation of Example OL-1. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.83 - 1.92 (m, 4 H), 1.95 -2.09 (m, 8 H), 2.11 -2.23 (m, 4 H), 2.75
-2.88
(m, 2 H), 2.92 (s, 1 H), 2.96 - 3.12 (m, 2 H), 3.97 (t, J=8.39 Hz, 2 H), 5.14
(dd,
J=17.70, 7.02 Hz, 2 H), 5.41 (s, 2 H), 6.91 -7.11 (m, J=49.75 Hz, 2 H), 7.14
(d,
J=7.93 Hz, 2 H), 7.44 - 7.68 (m, 12 H), 7.79 (d, J=8.54 Hz, 2 H), 7.84 - 8.02
(m,
J=9.16 Hz, 2 H), 10.21 (s, 1 H). Note: the signal of the imidazole NH was too
broad
to assign a chemical shift. LC (Cond. 1): RT = 1.78 min; LRMS: Anal. Calcd.
for
C46H51N803 (M+H)+ 763.41; found: 763.56. HRMS: Anal. Calcd. for C46H51N803
(M+H)+ 763.4084; found: 763.4067.
Examples OL-5 and OL-6
0 I N'---<ss
(40 H
0\
RA )--NH
Examples OL-5 to OL-6 were prepared as TFA salts by substituting the
respective acids for Cap-1 according to the same method described for Example
OL-
4.
Example 0 RT (LC-Cond.); MS data
R)L3S
Acid
OL-5 0 RT = 1.78 min. LC (Cond. 2); LCMS: Anal. Calcd.
Ph ..AsS for: C42H41N605 (M+H)+ 709.31; Found: 709.42
Ha- HRMS: Anal. Calcd. for: C42H41N605(M+H)+
Mandelic acid 709.3138; Found: 709.3130
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Example 0 RT (LC-Cond.); MS data
R)LsS
Acid
OL-6 0 RT = 1.96 min. LC (Cond. 2); LCMS: Anal. Calcd.
Ph
c.S' for: C46H471\1807(M+H)+ 823.36; Found: 823.51,
HNy(:) HRMS: Anal. Calcd. for: C46H471\1807(M+H)+
0 823.3568; Found: 823.3588
Cap-4
Example OL-7
0
N \ IF\11 N =
104 yce2irr---
H N 0
N---
Example OL-7a
HO
0 = 0
0
OH
To a solution of 4-(chloromethyl)benzoic acid (8.53 g, 50 mmol) and sodium
hydroxide (10 g, 0.25 mol) in water (40 mL) was added a solution of 4-
hydroxybenzoic acid (6.9 g, 50 mmol) and sodium hydroxide (6 g, 0.15 mol) in
water
(50 mL). The resulting mixture was heated to reflux temperature for 15 h and,
after
cooling to room temperature, it was filtered and acidified with a solution of
sulfuric
acid in water (1:1, 200 mL). The resulting precipitate was filtered off,
washed with
water and dried in vacuo. The off-white solid (9.3 g) was used without further
purification. 1H NMR (500 MHz, DMSO-d6) 6 ppm 5.27 (s, 1 H), 7.10 (d, J=8.85
Hz, 2 H), 7.56 (d, J=8.24 Hz, 2 H), 7.89 (d, J=8.85 Hz, 2 H), 7.96 (d, J=8.24
Hz, 2
H), 12.36 (br. s, 2 H). LC (Cond. 2), Note: The molecule did not ionize well
in the
LC/MS system and therefore an exact mass was not obtained.
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Example OL-7b
/
0
0 411 0 . 0
0
/
(Diazomethyl)trimethylsilane (15 mL, 30 mmol, 2 M in hexanes) was added
dropwise to a suspension of Example OL-7a (2.04 g, 7.5 mmol) in methanol (30
mL)
at 5 C (ice-water bath). When the addition was complete, the cooling bath was
removed and the mixture was stirred at ambient overnight. The solvent was
removed
under reduced pressure and the remaining residue was taken up in ethyl
acetate,
washed with water, sodium bicarbonate and brine, dried (MgSO4), filtered and
concentrated in vacuo. The resulting material was purified by flash
chromatography
(silica gel, 10% ethyl acetate/hexanes) to provide Example OL-7b as white
solid (1.5
g). 1H NMR (500 MHz, DMSO-d6) 6 ppm 3.80 (s, 3 H), 3.85 (s, 3 H), 5.29 (s, 2
H),
7.13 (d, J=8.85 Hz, 2 H), 7.59 (d, J=8.24 Hz, 2 H), 7.91 (d, J=9.16 Hz, 2 H),
7.98 (d,
J=8.24 Hz, 2 H). LC (Cond. 1): RT = 2.56 min; LRMS: Anal. Calcd. for C17H1605
(M+H)+ 301.11; found: 301.22.
Example OL-7c
001
. 0 . 0
CI
Example OL-7c was prepared from Example OL-7b, according to the same
procedure used for the preparation of Example OL-la. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 5.09 (s, 2 H), 5.18 (s, 2 H), 5.34 (s, 2 H), 7.15 (d, J=8.85 Hz, 2
H), 7.62 (d,
J=8.55 Hz, 2 H), 7.95 (d, J=9.16 Hz, 2 H), 8.00 (d, J=8.55 Hz, 2 H). LC (Cond.
1):
RT = 2.36 min; LRMS: Anal. Calcd. for C17H15C1203 (M+H)+ 337.04 found: 337.09.
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Example OL-7d
NH2
0 # 0 * 0
H2N
Example OL-7d was prepared from Example OL-7c, according to the same
procedure used for the preparation of Example OL-lb. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 4.49 (s, 2 H), 4.57 (s, 2 H), 5.38 (s, 2 H), 7.19 (d, J=8.85 Hz, 2
H), 7.66 (d,
J=8.24 Hz, 2 H), 8.00 (d, J=8.85 Hz, 2 H), 8.05 (d, J=8.24 Hz, 2 H), 8.45 (s,
6 H).
LC (Cond. 1): RT = 1.26 min; LRMS: Anal. Calcd. for C17H19N1203 (M+H)+ 299.14
found: 299.19.
Example OL-7e
Boc,
0 H .r9
N
140:1 0
0
C10 0
N
NLH 0
Boc
Example OL-7e was prepared from Example OL-7d, according to the same
procedure used for the preparation of Example OL-lc. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.30 - 1.47 (m, 18 H), 1.65 - 1.89 (m, 6 H), 2.01 - 2.20 (m, 2 H),
3.24 -
3.34 (m, 2 H), 3.34 - 3.43 (m, 2 H), 4.09 - 4.25 (m, 2 H), 4.44 - 4.68 (m, 4
H), 5.33 (s,
2 H), 7.13 (d, J=8.85 Hz, 2 H), 7.60 (d, J=8.24 Hz, 2 H), 7.99 (dd, J=20.29,
8.09 Hz,
4 H), 8.04 - 8.22 (m, J=31.74 Hz, 2 H). LC (Cond. 2): RT = 1.53 minutes, 97%
homogeneity index; LCMS: Anal. Calcd. for C37H49N409(M+1)+ 693.35; found:
693.32.
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Example OL-7f
2.,------
Boc HN /
\\I NHNBoc
Example OL-7f was prepared from Example OL-7e, according to the same
procedure used for the preparation of Example OL-1d. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 1.06 - 1.52 (m, 18 H), 1.69 -2.08 (m, 6 H), 2.08 - 2.30 (m, 2 H),
3.33 -
3.43 (m, 2 H), 3.52 (s, 2 H), 4.75 (s, 1 H), 4.83 (s, 1 H), 5.06 (s, 2 H),
6.92 - 7.10 (m,
J=7.63 Hz, 2 H), 7.20 - 7.33 (m, 1 H), 7.40 (d, J=7.63 Hz, 2 H), 7.46 (d,
J=8.55 Hz, 1
H), 7.64 (d, J=7.63 Hz, 2 H), 7.75 (d, J=7.93 Hz, 2 H), 11.47 - 12.18 (m, 2
H). LC
(Cond. 1): RT = 2.94 min; LRMS: Anal. Calcd. for C37H47N603 (M+H)+ 655.36
found: 655.39.
Example OL-7g
nH
\ NH
Example OL-7g was prepared from Example OL-7fe, according to the same
procedure used for the preparation of Example OL-le. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.93 -2.07 (m, 2 H), 2.13 -2.26 (m, 2 H), 2.40 -2.48 (m, 3 H), 3.31 -
3.52
(m, 5 H), 4.91 - 5.09 (m, 2 H), 5.21 (s, 2 H), 7.17 (d, J=8.85 Hz, 2 H), 7.57
(d, J=8.24
Hz, 2 H), 7.87 (d, J=8.85 Hz, 2 H), 7.93 (d, J=8.24 Hz, 2 H), 8.03 (d, J=16.79
Hz, 2
H), 9.78 (s, 1 H), 9.95 (s, 1 H), 10.31 (s, 2 H). Note: the signal of the
imidazole NH
was too broad to assign a chemical shift. LC (Cond. 2): RT = 0.62 minutes;
LCMS:
Anal. Calcd. for C27H31N60 (M+1)+ 455.26; found: 455.34.
Example OL-7
Example OL-7 was prepared from Example OL-7g according to the same
procedure used for the preparation of Example OL-1. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.90 (d, J=2.44 Hz, 2 H), 2.04 (d, J=3.66 Hz, 4 H), 2.21 (s, 2 H),
2.36 (d,
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J=1.83 Hz, 2 H), 2.42 - 2.46 (m, 4 H), 2.83 (br. s,4 H), 2.99 - 3.11 (m, 2 H),
3.98 (d,
J=7.02 Hz, 2 H), 5.18 (s, 4 H), 5.42 (d, J=8.55 Hz, 2 H), 7.08 - 7.28 (m, 3
H), 7.50 -
7.67 (m, 11 H), 7.73 (d, J=7.32 Hz, 2 H), 7.80 (d, J=7.93 Hz, 2 H), 7.85 -
7.98 (m,
J=10.38 Hz, 1 H). Note: the signal of the imidazole NH was too broad to assign
a
chemical shift. LC (Cond. 3): RT = 2.94 min; LRMS: Anal. Calcd. for C47H52N803
(M+H)+ 777.42 found: 777.51; HRMS: Anal. Calcd. for C47H53N803 (M+H)+
777.4241 found: 777.4265.
Example OL-8
H
,e
R \ = 0 ___________
NN
H 0
Example OL-8 was prepared as TFA salt by substituting the respective acid
for Cap-1 according to the same method described for Example OL-7.
Example 0 RT (LC-Cond.); MS data
R)Ls5
Acid
OL-8 0 RT = 3.70 min. LC (Cond. 4); LCMS: Anal. Calcd. for
Ph j=L
C47H49N807 (M+H) 837.37, Found. 837.54, HRMS.
HRy Anal. Calcd. for: C47F149N807 (M+H)+ 837.3724;
Found:
0 837.3715
Cap-4
Example OL-9
'N/
0 N IF\11 =
110
N 0
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Example OL-9a
¨0
/ \
¨ 0
¨
Palladium on carbon (10%, 300 mg) was added to a solution of dimethyl 4,4'-
(ethene-1,2-diy1)dibenzoate (2g, 6.76 mmol) in methanol (100 mL). The
suspension
was purged with N2, placed under 1 atm of H2 (balloon) and stirred at ambient
overnight. The mixture was then filtered through a plug of diatomaceous earth
(CELITEED) and the solvent was concentrated under reduced pressure. A white
solid
was recovered (1.42 g) and used without further purification. 1H NMR (500 MHz,
CDC13) 6 ppm 2.98 (s, 4 H), 3.90 (s, 6 H), 7.18 (d, J=7.32 Hz, 4 H), 7.93 (d,
J=7.32
Hz, 4 H). LC (Cond. 1): RT = 2.65 min; LRMS: Anal. Calcd. for C18H1904 (M+H)+
299.13 found: 299.21.
Example OL-9b
CI
.
0
1, 0 CI
Example OL-9b was prepared from Example OL-9a, according to the same
procedure used for the preparation of Example OL-la. 1H NMR (500 MHz, CDC13)
6 ppm 3.02 (s, 4 H), 4.67 (s, 4 H), 7.25 (d, J=8.24 Hz, 4 H), 7.86 (d, J=8.24
Hz, 4 H).
LC (Cond. 2): RT = 0.62 min; LCMS: Anal. Calcd. for C18H17C1202 (M+1)+ 335.06;
found: 335.12.
Example OL-9c
H2N
.
0 . 0
NH2
2 HCI
Example OL-9c was prepared from Example OL-9b, according to the same
procedure used for the preparation of Example OL-lb. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 3.03 (s, 4 H), 4.51 (s, 4 H), 7.43 (d, J=8.55 Hz, 4 H), 7.92 (d,
J=8.24 Hz, 4
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H), 8.50 (s, 6 H). LC (Cond. 3): RT = 1.90 min; LRMS: Anal. Calcd. for
C18H21N202 (M+H)+ 297.16 found: 297.24.
Example OL-9d
Boo\
0 Fi.r0
N
01 0
0
0
C7N
N*LH 0
Boc
Example OL-9d was prepared from Example OL-9c, according to the same
procedure used for the preparation of Example OL-1c. 1H NMR (500 MHz, CDC13)
6 ppm 1.38 - 1.56 (m, 18 H), 1.83 -2.00 (m, 4 H), 2.08 -2.38 (m, 4 H), 3.01
(s, 4 H),
3.32 - 3.65 (m, 4 H), 4.28 (br. s, 1 H), 4.39 (br. s, 1 H), 4.63 - 4.71 (m, 2
H), 4.72 -
4.84 (m, J=4.58 Hz, 2 H), 7.01 (s, 1 H), 7.18 - 7.33 (m, 4 H), 7.47 (s, 1 H),
7.87 (d,
J=7.93 Hz, 4 H). LC (Cond. 1): RT = 2.56 min; LRMS: Anal. Calcd. for
C38H51N408 (M+H)+ 691.37, found: 691.48.
Example OL-9e
n
c)Nr.õ-..N / ___________________ \ lik
Boo HN / _
Example OL-9e was prepared from Example OL-9d, according to the same
procedure used for the preparation of Example OL-1d. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.15/1.39(rotomers, s, 18 H), 1.75 - 1.92 (m, 3 H), 1.96 (s, 3 H),
2.09 -
2.27 (m, J=32.04 Hz, 2 H), 2.81 -2.90 (m, 4 H), 3.31 -3.40 (m, 2 H), 3.52 (s,
2 H),
4.66 -4.88 (m, J=35.40 Hz, 2 H), 7.14 (d, J=7.02 Hz, 4 H), 7.21 (s, 1 H), 7.31
- 7.41
(m, 1 H), 7.51 (d, J=6.41 Hz, 1 H), 7.61 (d, J=7.93 Hz, 3 H), 11.62- 12.08 (m,
2 H).
LC (Cond. 1): RT = 2.20 min; LRMS: Anal. Calcd. for C38H49N604 (M+H)+ 653.38,
found: 653.47.
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Example OL-9f
=
HN
H
Example OL-9f was prepared from Example OL-9e, according to the same
procedure used for the preparation of Example OL-le. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.92 - 2.05 (m, 2 H), 2.11 - 2.25 (m, 2 H), 2.40 -2.47 (m, 4 H),
2.89 - 3.03
(m, 4 H), 3.32 - 3.39 (m, 2 H), 3.40 - 3.49 (m, 2 H), 5.02 (t, J=7.78 Hz, 2
H), 7.32 (d,
J=8.24 Hz, 4 H), 7.80 (d, J=8.24 Hz, 4 H), 9.91 (s, 2 H), 10.32 (s, 2 H).
Note: the
signal of the imidazole NH was too broad to assign a chemical shift. LC (Cond.
5):
RT = 2.52 min; LRMS: Anal. Calcd. for C28F133N6 (M+H)+ 453.28, found: 453.31.
Example OL-9
Example OL-9 was prepared from Example OL-9f, according to the same
procedure used for the preparation of Example OL-1. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 1.82 - 1.95 (m, 2 H), 1.97 -2.10 (m, 4 H), 2.15 -2.27 (m, J=4.58 Hz,
2 H),
2.28 - 2.45 (m, 4 H), 2.72 - 2.88 (m, 4 H), 2.94 (s, 5 H), 2.98 - 3.07 (m, 2
H), 3.97 (t,
J=9.77 Hz, 2 H), 5.17 (d, J=6.10 Hz, 2 H), 5.41 (s, 2 H), 7.12 (s, 1 H), 7.38
(d, J=7.32
Hz, 4 H), 7.51 - 7.62 (m, 9 H), 7.69 (d, J=7.93 Hz, 4 H), 7.93 (s, 2 H). Note:
the
signal of the imidazole NH was too broad to assign a chemical shift. LC (Cond.
5):
RT = 2.87 min; LRMS: Anal. Calcd. for C48H55N802 (M+H)+ 775.44 found: 775.51;
HRMS: Anal. Calcd. for C48H55N802 (M+H)+ 775.4448 found: 775.4454.
Examples OL-10 to OL-13
H N
0
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Examples OL-10 to OL-13 were prepared as TFA salts by substituting the
respective acids for Cap-1 according to the same method described for Example
OL-
9.
Example 0 RT (LC-Cond.); MS data
R)LsS
Acid
OL- 10 0 RT = 2.64 min. LC (Cond. 4); LCMS: Anal. Calcd. for:
Ph .)..LsS C44H45N604 (M+H)+ 721.35; Found: 721.42, HRMS:
Ho7. Anal. Calcd. for C44H45N604 (M+H)+ 721.3502;
Found:
Mandelic acid 721.3518
OL-11 0 RT = 2.88 min. LC (Cond. 4); LCMS: Anal. Calcd. for:
Ph .).L.c.0 C48H5iN806 (M+H)+ 835.39; Found: 835.49 HRMS:
FliCiy Anal. Calcd. for: C48H50N806(M+H)+ 835.3932;
Found:
0 835.3939
Cap-4
OL- 12 0 RT = 4.20 min. LC (Cond. 4); LCMS: Anal. Calcd. for:
Ph sss'- C501-157N1004 (M+H)+ 861.45; Found: 861.50,
HRMS:
HNr0 Anal. Calcd. for: C501457N1004(M+H)+861.4564;
NH Found: 861.4562
Cap-46
OL-13 0 RT = 5.11 min. LC (Cond. 4); LCMS: Anal. Calcd. for:
Ph,C56H64N1004 (M+H)+ 941.52; Found: 941.71, HRMS:
HNr0 Anal. Calcd. for: C56H651\11004 (M+H)+ 941.5190;
NH
Found: 941.5162
Cap-48
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Example OL-14
N N
n 10 , õ
N hi
0
-N¨
/ ¨N*
\
Example OL-14a
0 0
Me0 0 0 OMe
0
Triethyl silane (5.84 mL, 36.55 mmol) was added dropwise to a solution of
methyl 4-formylbenzoate (5 g, 30.46 mmol) and bismuth tribromide (0.273 g,
0.61
mmol) in acetonitrile (75 mL) and the opaque solution was stirred at ambient
for 15
min. Volatiles were removed under reduced pressure and the remaining residue
was
taken up in ethyl acetate. The organic layer was then washed with water and
brine,
dried (MgSO4), filtered and concentrated in vacuo. The recovered solid was
then
submitted to flash chromatography (silica gel; 10-20% ethyl acetate/hexanes)
to
provide OL-14a (3.05 g). 1H NMR (500 MHz, CDC13) 6 ppm 3.84 (s, 6 H), 4.65 (s,
4
H), 7.51 (d, J=8.24 Hz, 4 H), 7.95 (d, J=8.24 Hz, 4 H). LC (Cond. 2): RT =
1.68 min;
LCMS: Anal. Calcd. for C18I-11905 (M+H)+ 315.12; found: 315.26.
Example OL-14b
0 0
CI el 0 110 CI
Example OL-14b was prepared from Example OL-14a, according to the same
procedure used for the preparation of Example OL-la. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 4.67 (s, 4 H), 5.19 (s, 4 H), 7.55 (d, J=8.24 Hz, 4 H), 7.98 (d,
J=8.24 Hz, 4
H). LC (Cond. 5): RT = 3.85 min; LRMS: Anal. Calcd. for C18H17C1203 (M+H)+
351.05, found: 351.06.
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Example OL-14c
0 0
NH: el 0 NH2
2 HCI
Example OL-14c was prepared from Example OL-14b, according to the same
procedure used for the preparation of Example OL-lb. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 4.55 (s, 4 H), 4.69 (s, 4 H), 7.57 (d, J=8.24 Hz, 4 H), 8.02 (d,
J=8.55 Hz, 4
H), 8.53 (s, 6 H). LC (Cond. 1): RT = 1.29 min; LRMS: Anal. Calcd. for
C18H21N203 (M+H)+ 313.15, found: 313.21.
Example OL-14d
0 0
c-jr H H
I
Boc 0 I 0 I. 0 Boc
Example OL-14d was prepared from Example OL-14c, according to the same
procedure used for the preparation of Example OL-lc. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.34/1.40 (rotomers, s, 18 H), 1.70 - 1.92 (m, 6 H), 1.99 - 2.20 (m,
2 H),
3.24 - 3.33 (m, 2 H), 3.35 - 3.44 (m, 2 H), 4.11 -4.25 (m, 2 H), 4.50 - 4.65
(m, 4 H),
4.66 (s, 4 H), 7.53 (d, J=8.24 Hz, 4 H), 7.99 (d, J=7.93 Hz, 4 H), 8.09 - 8.23
(m, 2 H).
LC (Cond. 1): RT = 2.49 min; LRMS: Anal. Calcd. for C38H51N409 (M+H)+ 707.36
found: 707.50.
Example OL-14e
HN NH
N I
N N
Boc 0 Boc
Example OL-14e was prepared from Example OL-14d, according to the same
procedure used for the preparation of Example OL-1d. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.14/ 1.39 (rotomers, s, 18 H), 1.76 - 2.06 (m, 6 H), 2.08 -2.30 (m,
2 H),
3.31 - 3.41 (m, 2 H), 3.52 (s, 2 H), 4.49 (s, 4 H), 4.70 -4.78 (m, 1 H), 4.79 -
4.90 (m,
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1 H), 7.29/7.35 (rotomers, d, J=7.93 Hz, 4 H), 7.40 - 7.54 (m, 2 H), 7.71/761
(rotomers, d, J=7.93 Hz, 4 H), 11.72 - 12.19 (m, 2 H). LC (Cond. 1): RT = 2.14
min;
LRMS: Anal. Calcd. for C38H49N605 (M+H)+669.38, found: 669.53.
Example OL-14f
HN NH
NH N SO . N HN
0
Example OL-14f was prepared from Example OL-14e, according to the same
procedure used for the preparation of Example OL-le. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.94 - 2.06 (m, 2 H), 2.11 -2.25 (m, 2 H), 2.38 - 2.48 (m, 4 H),
3.32 - 3.50
(m, 4 H), 4.60 (s, 4 H), 4.92 - 5.06 (m, 2 H), 7.48 (d, J=7.93 Hz, 4 H), 7.89
(d, J=7.93
Hz, 4 H), 8.04 (s, 2 H), 9.78 (s, 2 H), 10.28 (s, 2 H). Note: the signal of
the
imidazole NH was too broad to assign a chemical shift. LC (Cond. 1): RT =1.69
min; LRMS: Anal. Calcd. for C28H33N60 (M+H)+ 469.27 found: 469.34.
Example OL-14
Example OL-14 was prepared from Example OL-14f, according to the same
procedure used for the preparation of Example OL-1. 1H NMR (500 MHz, DMSO-
d6) 6 ppm 1.82 -2.11 (m, 6 H), 2.18 (d, J=6.10 Hz, 2 H), 2.31 -2.47 (m, 4 H),
2.98 -
3.10 (m, 2 H), 3.92 - 4.07 (m, 2 H), 4.59 (s, 4 H), 5.10 - 5.22 (m, 2 H), 5.42
(s, 2 H),
7.07 - 7.13 (m, 2 H), 7.40 - 7.51 (m, 4 H), 7.54 - 7.67 (m, 10 H), 7.75 (d,
J=7.32 Hz,
4 H). Note: the signal of the imidazole NH was too broad to assign a chemical
shift.
The signals for the Me groups of the cap fell underneath the solvent peak. LC
(Cond.
4): RT = 1.83 min; LRMS: Anal. Calcd. for C48H55N803 (M+H)+ 791.44; found:
791.60. HRMS: Anal. Calcd. for C48H55N803 (M+H)+ 791.4397; found: 791.4406.
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Examples OL-15 to OL-19
N N
0
Examples OL-15 to OL-19 were prepared as TFA salts by substituting the
respective acids for Cap-1 according to the same method described for Example
OL-
14.
Example 0 RT (LC-Cond.); MS data
R)L3S
Acid
OL- 15 0 RT = 2.86 min. LC (Cond. 4); LCMS: Anal. Calcd.
Ph .).L.5.5' for: C44H45N605 (M+H)+ 737.34; Found: 737.38,
HO HRMS: HRMS: Anal. Calcd. for: C44H45N605 (M+H)+
Mandelic acid 737.3451; Found: 737.3452
OL- 16 0 RT = 3.45 min. LC (Cond. 4); LCMS: Anal. Calcd.
for:
Ph C48H51N807 (M+H)+ 851.39; Found: 851.40, HRMS:
FIR yc) Anal. Calcd. for: C48H51N807 (M+H)+ 851.3881;
0 Found: 851.3910
Cap-4
OL-17 0 RT = 1.16 min. LC (Cond. 2); LCMS: Anal. Calcd.
for:
Ph As'- C48H53N1005 (M+H)+ 849.42; Found: 849.61, HRMS:
HN0 Anal. Calcd. for: C481-153N1005 (M+H)+ 849.4200;
NH
Found: 849.4182
Cap-45a
OL-18 0 RT = 1.24 min. LC (Cond. 2); LCMS: Anal. Calcd.
Ph/for: C501-156N1005 (M+H)+ 877.45; Found: 877.70,
HN0 HRMS: Anal. Calcd. for: C50H571\11005 (M+H)+
NH 877.4513; Found: 877.4506
Cap-46
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Example 0 RT (LC-Cond.); MS data
Acid
OL-19 0 RT = 1.43 min. LC (Cond. 2); LCMS: Anal. Calcd.
Ph .Ass! for: C56H651\11005 (M+H)+ 957.51; Found: 957.84,
HN 0
HRMS: Anal. Calcd. for: C56H651\11005 (M+H)+
NH 957.5139; Found: 957.5142
Cap-48
Example OL-20
¨1\1
C 0
0 H
/
Example OL-20a
0
0
0 0
0
Sodium hydride (0.76 g, 18.95 mmol, 60% wt) was slowly added to a solution
of methyl 4-(hydroxymethyl)benzoate (3 g, 18.05 mmol) in dimethylformamide (25
mL). The resulting dark blue solution was stirred at ambient for 15 min and
methyl
3-(bromomethyl)benzoate (3.76g, 16.41 mmol) was added in one portion. The
resulting yellow solution was stirred at ambient for 1 h and then the
volatiles were
removed under reduced pressure. The residue was taken up in ethyl acetate and
washed with water and brine, dried (Mg504), filtered and concentrated in
vacuo.
The residue was submitted to flash chromatography (silica gel; 20 % ethyl
acetate/hexanes) to provide Example OL-20a as a clear oil that solidified upon
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standing (2.49 g). 1H NMR (300 MHz, CDC13) 6 ppm 3.91 (d, J=1.83 Hz, 6 H),
4.61
(d, J=2.56 Hz, 4 H), 7.39 - 7.49 (m, 3 H), 7.54 - 7.61 (m, 1 H), 7.93 - 8.00
(m, 1 H),
7.99 - 8.07 (m, 3 H). LC (Cond. 1): RT =2.56 min; LRMS: Anal. Calcd. for
C18H1905 (M+H)+ 315.12 found: 315.22.
Example OL-20b
0
CI
el 0 110
CI
0
Example OL-20b was prepared from Example OL-20a, according to the same
procedure used for the preparation of Example OL-la. 1H NMR (500 MHz, CDC13)
6 ppm 4.65 (d, J=4.58 Hz, 4 H), 4.69 (d, J=3.36 Hz, 4 H), 7.47 - 7.54 (m,
J=7.63,
7.63 Hz, 3 H), 7.62 (d, J=7.63 Hz, 1 H), 7.89 (d, J=7.93 Hz, 1 H), 7.93 - 8.00
(m, 3
H). LC (Cond. 1): RT =2.34 min; LRMS: Anal. Calcd. for C18H17C1203 (M+H)+
351.05 found: 351.12.
Example OL-20c
0
H2N
101 0 0
NH2
2 HCI 0
Example OL-20c was prepared from Example OL-20b, according to the same
procedure used for the preparation of Example OL-lb. 1H NMR (300 MHz, DMSO-
d6) 6 ppm 4.57 (d, J=2.93 Hz, 4 H), 4.68 (d, J=5.85 Hz, 4 H), 7.52 - 7.67 (m,
J=8.60,
8.60 Hz, 3 H), 7.73 (d, J=7.68 Hz, 1 H), 7.90 - 8.10 (m, 4 H), 8.53 (s, 6 H).
LC
(Cond. 1): RT =1.32 min; LRMS: Anal. Calcd. for C18H21N203 (M+H)+ 313.15
found: 313.24.
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Example OL-20d
0
9rH
0 Boc
Bo c 0 0 N NI
0
Example OL-20d was prepared from Example OL-20c, according to the same
procedure used for the preparation of Example OL-1c. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 1.39/1.34 (rotomers, s, 18 H), 1.69 - 1.95 (m, 6 H), 2.00 -2.18 (m,
J=23.80
Hz, 2 H), 3.23 -3.33 (m, 2 H), 3.34 - 3.43 (m, 2 H), 4.11 -4.23 (m, 2 H), 4.50
-4.63
(m, 4 H), 4.65 (d, J=6.71 Hz, 4 H), 7.48 - 7.56 (m, 3 H), 7.66 (d, J=7.63 Hz,
1 H),
7.89 - 8.02 (m, 4 H), 8.06 - 8.25 (m, 2 H). LC (Cond. 1): RT =2.53 min; LRMS:
Anal. Calcd. for C38H51N409 (M+H)+ 707.36 found: 707.44.
Example OL-20e
BOG H
N
0-4N el 0
N
I ___________________________________________________ J
N N
H Boo/
Example OL-20e was prepared from Example OL-20d, according to the same
procedure used for the preparation of Example OL-1d. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 1.14/1.39 (rotomers, s, 18 H), 1.76 -2.07 (m, 6 H), 2.08 -2.29 (m, 2
H),
3.31 - 3.40 (m, 2 H), 3.52 (br. s, 2 H), 4.44 - 4.59 (m, J=8.55 Hz, 4 H), 4.75
(br. s, 1
H), 4.83 (br. s, 1 H), 7.08 - 7.23 (m, 2 H), 7.30 (d, J=7.63 Hz, 2 H), 7.34 -
7.50 (m, 2
H), 7.53 -7.67 (m, 1 H), 7.68 - 7.79 (m, 3 H), 11.70- 12.22 (m, 2 H). LC
(Cond. 1):
RT =2.19 min; LRMS: Anal. Calcd. for C38H48N605 (M+H)+ 669.39, found: 669.40.
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Example OL-20f
HH
N N
------ I
L- N ei
0 11101
I > _________________________________________________ \
N\ .,,--.3
N N
H H
Example OL-20f was prepared from Example OL-20e, according to the same
procedure used for the preparation of Example OL-le. 1H NMR (500 MHz, DMS0-
D6) 6 ppm 1.94 -2.08 (m, 2 H), 2.12 - 2.24 (m, 2 H), 2.39 -2.48 (m, J=5.19 Hz,
4
H), 3.27 - 3.49 (m, 4 H), 4.61 (d, J=4.88 Hz, 4 H), 4.94 - 5.09 (m, J=7.93 Hz,
2 H),
7.38 (d, J=7.32 Hz, 1 H), 7.45 - 7.53 (m, 3 H), 7.83 (d, J=7.63 Hz, 1 H), 7.90
(d,
J=7.02 Hz, 3 H), 8.07 (s, 2 H), 9.86 (br. s, 2 H), 10.33 (br. s, 2 H). Note:
the signal
of the imidazole NH was too broad to assign a chemical shift. LC (Cond. 1): RT
=
1.75 min; LRMS: Anal. Calcd. for C28H32N60 (M+H)+ 469.27, found: 469.23.
Example OL-20
Example OL-20 was prepared from Example OL-20f, according to the same
procedure used for the preparation of Example OL-1. 1H NMR (500 MHz, DMS0-
d6) 6 ppm 1.84 - 1.95 (m, J=5.19 Hz, 2 H), 1.95 -2.09 (m, J=8.24 Hz, 4 H),
2.15 -
2.28 (m, 2 H), 3.02 (q, J=7.73 Hz, 2 H), 3.93 -4.05 (m, 2 H), 4.61 (br. s, 4
H), 5.12 -
5.22 (m, 2 H), 5.42 (br.s, 2 H), 7.12 (br. s, 1 H), 7.39 (s, 1 H), 7.45 - 7.51
(m, J=7.32
Hz, 3 H), 7.53 - 7.64 (m, 9 H), 7.66 - 7.82 (m, 4 H), 7.96 (s, 2 H). Note: the
signal of
the imidazole NH was too broad to assign a chemical shift. The signals for the
Me
groups of the cap fell underneath the solvent peak. LC (Cond. 4): RT = 1.93
min;
LRMS: Anal. Calcd. for C48H55N803 (M+H)+ 791.44; found: 791.57. HRMS: Anal.
Calcd. for C48H55N803 (M+H)+. 791.4397; found: 791.4373.
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Examples OL-21 to OL-25
\r0
N HN
N
0 N
N)
NH
0\ R
Examples OL-21 to OL-25 were prepared as TFA salts by substituting the
respective acids for Cap-1 according to the same method described for Example
OL-
20.
Example 0 RT (LC-Cond.); MS data
R)L3S
Acid
OL-2 1 0 RT = 2.97 min. LC (Cond. 4); LCMS: Anal. Calcd.
for:
Ph :).LsS C44H45N605 (M+H)+ 737.34; Found: 737.46 HRMS:
1-12-8 Anal. Calcd. for: C44H45N605 (M+H)+ 737.3451;
Found:
Mandelic acid 737.3459
OL-22 0 RT = 3.59 min. LC (Cond. 4); LCMS: Anal. Calcd.
for:
Ph j=L
sS C48H5iN807 (M+H) 85 1.3 8, Found. 851.50, HRMS.
F-1-11y Anal. Calcd. for: C481-151N807 (M+H)+ 851.3881;
Found:
0 851.3893
Cap-4
OL-23 0 RT = 1.15 min. LC (Cond. 2); LCMS: Anal. Calcd.
for:
Ph/C48H52N1005 (M+H)+ 849.42; Found: 849.46, HRMS:
HN0 Anal. Calcd. for: C48H53N1005 (M+H)+ 849.4200;
Found:
NH
849.4185
Cap-45a
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Example 0 RT (LC-Cond.); MS data
R)LsS'
Acid
OL-34 0 RT = 1.24 min. LC (Cond. 2); LCMS: Anal. Calcd.
for:
Ph sss'- C501-156N1005 (M+H)+ 877.45 Found: 877.69,
HRMS:
HN
Anal. Calcd. for: C50F1571\11005(M+H)+ 877.4513; Found:
r NH 877.4510
Cap-46
OL-25 o RT = 4.89 min. LC (Cond. 4); LCMS: Anal. Calcd.
for:
Ph ss'= C56H64N1005(M+H)+ 957.51; Found: 957.82, HRMS:
HN
Anal. Calcd. for: C56H651\11005 (M+H)+ 957.5139; Found:
aNH 957.5150
Cap-48
Example D-1
= H 0
N
0
N
N\ =
N H
CNI: 0
0 2 T F A
0)L IF1
=
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Example D-1, step a
Br
0 Boc
NN
H
0
N,N-Diisopropylethylamine (18 mL, 103.3 mmol) was added dropwise, over
15 minutes, to a heterogeneous mixture of N-Boc-L-proline (7.14 g, 33.17
mmol),
HATU (13.32 g, 35.04 mmol), the HC1 salt of 2-amino-1-(4-bromophenyl)ethanone
(8.13 g, 32.44 mmol), and DMF (105 mL), and stirred at ambient condition for
55
minutes. Most of the volatile component was removed in vacuo, and the
resulting
residue was partitioned between ethyl acetate (300 mL) and water (200 mL). The
organic layer was washed with water (200 mL) and brine, dried (MgSO4),
filtered,
and concentrated in vacuo. A silica gel mesh was prepared from the residue and
submitted to flash chromatography (silica gel; 50-60 % ethyl acetate/hexanes)
to
provide D-la as a white solid (12.8 g). 1H NMR (500 MHz, DMSO-d6) 6 ppm
1.40/1.34 (two app br s, 9 H), 1.90-1.70 (m, 3 H), 2.18-2.20 (m, 1 H), 3.30-
3.23 (m, 1
H), 3.43-3.35 (m, 1 H), 4.22-4.12 (m, 1 H), 4.53 (dd, J=18.1, 5.6, 1 H), 4.61
(dd,
J=18.3, 5.7, 1 H), 7.75 (br d, J=8.6, 2 H), 7.92 (br d, J=8.0, 2 H), 8.25-8.14
(m, 1H).
LC (Cond. 7): RT = 1.70 min; LRMS: Anal. Calcd. for (M+Na)+ C18H23BrN2Na04:
433.07 found 433.09.
Example D-1, step b
H n
Br
\ Boc
A mixture of D- 1 a (12.8 g, 31.12 mmol) and NH40Ac (12.0 g, 155.7 mmol)
in xylenes (155 mL) was heated in a sealed tube at 140 C for 2 hours. The
volatile
component was removed in vacuo, and the residue was partitioned carefully
between
ethyl acetate and water, whereby enough saturated NaHCO3 solution was added so
as
to make the pH of the aqueous phase slightly basic after the shaking of the
biphasic
system. The layers were separated, and the aqueous layer was extracted with
additional ethyl acetate. The combined organic phase was washed with brine,
dried
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(MgSO4), filtered, and concentrated in vacuo. The resulting material was
recrystallized from ethyl acetate/hexanes to provide two crops of imidazole D-
lb as a
light-yellow, dense solid, weighing 5.85 g. The mother liquor was concentrated
in
vacuo and submitted to a flash chromatography (silica gel; 30% ethyl
acetate/hexanes) to provide an additional 2.23 g of Example D-lb. 1H NMR (500
MHz, DMSO-d6) 6 ppm 1.40/1.15 (app br s, 9 H), 2.30-1.75 (m, 4 H), 3.36 (m, 1
H),
3.52 (app br s, 1 H), 4.86-4.70 (m, 1 H), 7.72-7.46/7.28 (m, 5 H),
12.17/11.92/11.86
(m, 1 H). LC (Cond. 7): RT = 1.71 min; LRMS: Anal. Calcd. for [M+H]+
C18H23BrN302: 392.10; found 391.96; HRMS: Anal. Calcd. for [M+H]+
C18H23BrN302: 392.0974; found 392.0959.
The optical purity of the two samples of Example D-lb was assessed
according to the chiral HPLC conditions noted below (ee > 99% for the combined
crops; ee = 96.7% for the sample from flash chromatography):
Column: Chiralpak AD, 10 um, 4.6 x 50 mm
Solvent: 2% ethanol/heptane (isocratic)
Flow rate: 1 mL/min
Wavelength: either 220 or 254 nm
Relative retention time: 2.83 minutes (R) , 5.34 minutes (S).
Example D-1, step c
--\...¨ H
0 7
'..'.--'.
µ I 3 . NIi N
-2- d \ N Boc
Pd(Ph3P)4 (469 mg, 0.41 mmol) was added to a pressure tube containing a
mixture of D-lb (4.01 g, 10.22 mmol), bis(pinacolato)diboron (5.42 g, 21.35
mmol),
potassium acetate (2.57 g, 26.21 mmol) and 1,4-dioxane (80 mL). The pressure
tube
was purged with nitrogen, capped and heated in an oil bath at 80 C for 16.5
hours.
The reaction mixture was filtered and the filtrate was concentrated in vacuo.
The
crude residue was partitioned carefully between CH2C12 (150 mL) and an aqueous
medium (50 mL water and 10 mL saturated NaHCO3 solution). The aqueous layer
was extracted with CH2C12, and the combined organic phase was dried (Mg504),
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filtered, and concentrated in vacuo. The resulting material was purified with
flash
chromatography (sample was loaded with eluting solvent; 20-35% ethyl
acetate/CH2C12) to provide Example D-lc, contaminated with pinacol, as an off-
white dense solid; the relative mole ratio of Example D-lc to pinacol was
about 10:1
(H NMR). The sample weighed 3.925 g after ¨2.5 days exposure to high vacuum.
1H NMR (500 MHz, DMSO-d6) 6 ppm 1.45-1.10 (m, 21H), 2.27-1.77 (m, 4H), 3.36
(m, 1H), 3.52 (app br s, 1H), 4.86-4.70 (m, 1H), 7.79-7.50/ 7.34-7.27 (m, 5H),
12.22/11.94/11.87 (m, 1H). LC (Cond. 7): RT = 1.64 min; LRMS: Anal. Calcd. for
[M+H]+ C24H35BN304: 440.27; found 440.23.
Example D-1, step d
>INO
,-,
us..;11.44)
HN \ 41 = N,_
= \ NH
N
elNr0
0(...õ..
Tetrakistriphenylphosphine palladium (17.5 mg, 0.015 mmol) was added in
one portion to a stirred suspension of D-lc (320 mg, 0.85 mmol), 1,4-
diiodobenzene
(100 mg, 0.30 mmol) and sodium bicarbonate (180 mg, 2.18 mmol) in
dimethoxyethane (2.4 mL) and water (7 mL) and heated to 90 C for 2 h. The
reaction was diluted with ethyl acetate and washed with saturated sodium
bicarbonate. The organic layer was dried over sodium sulfate, filtered and
concentrated in vacuo. The filtrate was taken up in methanol, filtered through
a
nylon syringe filter and then purified by preparative HPLC to provide D-ld as
a
white solid (101.1 mg, 20% yield). 1H NMR (500 MHz, DMSO-d6) 6 ppm 1.17 (br s,
9 H), 1.41 (br s, 9 H), 1.80 - 2.06 (m, 4 H), 2.13 -2.32 (m, 4 H), 3.35 - 3.43
(m, 2 H),
3.56 (br. s., 2 H), 4.80 (br s, 2 H), 7.54 (d, J=10.07 Hz, 2 H), 7.73 (d,
J=7.63 Hz, 4
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H), 7.79 (s, 4 H), 7.83 (d, J=8.24 Hz, 4 H). LC (Cond. 6): RT = 2.55 min;
HRMS:
Anal. Calcd. for (M+H)+C42H49N604 701.3815; found: 701.3790.
Example D-1, step e
_17,44)
= \1\1*---N H
D-le was prepared from D-1d, according to the same procedure used for the
preparation of OL-le except that methanol (1 mL) was used instead of
dichloromethane. This gave D-le as a tan solid and as a hydrochloride salt
(101.5
mg, 96 % yield). 1H NMR (500 MHz, DMSO-d6) 6 ppm 1.93 - 2.30 (m, 9 H), 2.32 -
2.44 (m, 3 H), 4.69 - 4.86 (m, 2 H), 7.68 - 7.87 (m, 12 H), 7.90 (d, J= 8.24
Hz, 2 H),
9.48 - 9.75 (m, 2 H). LC (Cond. 6): RT = 1.99 min; HRMS: Anal. Calcd. for
(M+H)+ C32H33N6 501.2767; found: 501.2753.
Example D-1
Example D-1 (TFA salt) was prepared from D-le, according to the same
procedure used for the preparation of Example OL-1. This gave Example D-1 as a
tan solid (37.8 mg, 54 % yield). 1H NMR (500 MHz, DMSO-d6) 6 ppm 2.05 (d,
J=8.55 Hz, 8 H), 3.49 - 3.57 (m, 10 H), 3.94 (br s, 2 H), 5.52 (d, J=7.63 Hz,
2 H),
7.31 - 7.47 (m, 10 H), 7.72 (d, J=7.02 Hz, 2 H), 7.87 - 8.01 (m, 12 H). LC
(Cond. 6):
RT = 2.41 min; HRMS: Anal. Calcd. for (M+H)+ C52H51N806 883.3932; found:
883.3947.
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Example D-2
410NN
HN =
NN,
Example D-2 (TFA salt) was prepared from pyrrolidine D-le, according to
the same procedure used for the preparation of Example OL-1. This gave Example
D-2 as a light yellow solid (15.4 mg, 38 % yield). 1H NMR (500 MHz, DMSO-d6) 6
ppm 2.50-1.70 (m, 14 H), 2.98-2.62 (m, 6 H), 4.05-2.98 (m, 4 H), 5.79-5.12 (m,
4 H),
8.20-7.10 (m, 24 H), 10.25 (br s, 2 H). Note: The signal of the imidazole NH
was too
broad to assign a chemical shift. LC (Cond. 6): RT = 2.08 min; HRMS: Anal.
Calcd.
For (M+H)+ C52H55N802823.4448; found: 823.4467.
Example D-3
o 0,
1-N 0
\\``
H
--05)LHN\
Example D-3, step a
Br &Ai
Br
A solution of bromine (683 L, 13.33 mmol) in acetic acid (7 mL) was added
dropwise to a cold (10 C) solution of 6-bromo-3,4-dihydronaphthalen-1(21])-
one
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(purchased from J& W PharmLab, LLC) (3.00 g, 13.33 mmol) and 48% hydrogen
bromide (20 [IL, 13.33 mmol) in acetic acid (120 mL). The mixture was allowed
to
warm up to rt after the addition was complete and allowed to stir at rt for 1
h before it
was diluted with dichloromethane and washed with water (3x), saturated sodium
bicarbonate solution, dried over anhydrous sodium sulfate and concentrated.
There
was isolated D-3a (4.19 g, 97% yield) as a colorless oil which solidified on
standing
under high vacuum to a white solid. This material was used without further
purification. 1FINMR (500 MHz, DMSO-d6) 6 ppm 7.85 (1 H, d, J=8.5 Hz), 7.71 (1
H, s), 7.62 (1 H, dd, J=8.5, 1.8 Hz), 5.06 (1 H, dd, J=6.1, 3.7 Hz), 2.93 -
3.15 (2 H,
m), 2.55 - 2.64 (1 H, m), 2.32 - 2.42 (1 H, m). RT = 2.67 min (Cond. 6);
LC/MS:
Anal. Calcd. for [M+H]+ C10H979Br20: 304.90; found: 304.91.
Examples D-3b1 and D-3b2 (step b)
D-3b1 and D-3b2 were prepared from D-3a and the appropriate Boc-protected
prolines according to the same procedure used for the preparation of Example
M3,
step g.
RT = 3.00 and 3.19
min (Cond. 3);
D-3b1 LC/MS: Anal.
I.Dc
Br u Oer _
Calcd. for [M+Na]+
o)CO
0 79
C20H25 BrN05:
460.08; found:
460.10.
RT = 2.99 and 3.17
u I.Dc
Br 40, _ min (Cond. 3),
D-3b2 0 H LCMS: Calcd for
0
H C21H2579BrN05
[M+Na]+ 472.07;
found: 472.10
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Examples D-3c1 and D-3c2 (step c)
D-3c1 and D-3c2 were prepared from D-3b1 and D-3b2 respectively
according to the same procedure used for the preparation of Example M3, step
h.
1H NMR (500 MHz, Me0D, imidazole
N-H missing) 6 ppm 7.32 - 7.43 (3 H, m),
4.86 -4.97 (1 H, m), 3.63 - 3.74 (1 H, m),
3.47 - 3.57 (1 H, m), 2.98 - 3.07 (2 H,
Br 4040
m), 2.83 (2 H, d, J=8.2 Hz), 2.25 - 2.45
D-3c1 (1 H, m), 2.00 - 2.11 (2 H, m), 1.93 _
Boc-N 1.99 (1 H, m), 1.25 - 1.48 (9 H, 2s).
RT =
From D-3b1 2.24 min (Cond. 3); LC/MS: Anal.
Calcd. for [M+H]+ C201-12579BrN302:
418.13; found: 418.10. HRMS: Anal.
Calcd. for [M+H]+ C211-12579BrN302:
430.1125; found 430.1124.
RT = 2.28 min (Cond. 3); LC/MS: Anal.
Br 00Calcd. for [M+H]+ C21H2579BrN302:
D-3c2 HN 430.11; found: 430.16.
noc-N
HRMS: Anal. Calcd. for [M+H]+
-n NT rµ A
l.21112579I 1GJ; found
\L
From D-3b2
430.1123.
Br 400
Boc-N RT = 2.06 min (Cond. 3); LC/MS: Anal.
D-3c3
Calcd for C21H2381BrN302 (M+H)+
Prepared from D-3c2
430.10; found: 429.98.
according to the same
procedure used for the
preparation of D-3g1
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Example D-3d1 (step d)
Si
BOG-N3
Ethynyltrimethylsilane (0.59 mL, 4.25 mmol) was added to a solution of D-lb
(1.5 g, 3.82 mmol), triphenylphosphine (0.20 g, 0.77 mmol), diethylamine (4.25
mL,
40.70 mmol), copper (I) iodide (40 mg, 0.21 mmol) and trans-dichloro(bis-
triphenylphosphine)palladium (II) (149 mg, 0.21 mmol) in dry DMF (1.4 mL) at
rt in
a microwave vessel. The vessel was capped and irradiated for 25 min at 120 C.
Two identical 1.5 g reactions were run in tandem. The reaction mixtures were
diluted with ether and ethyl acetate, combined and shaken with 0.1N HC1. After
standing for 20 min, the suspension was suction-filtered and the pad was
washed with
ether and ethyl acetate. The organic phase was then separated, washed with
brine,
dried over sodium sulfate, and concentrated. There was isolated the crude
product
(4.2 g) as a brownish-red foam which was taken up in dichloromethane and added
directly to a Thompson 110 g silica gel column. Gradient elution of the
residue with
20% ethyl acetate in dichloromethane to 100% ethyl acetate furnished D-3d1
(2.8 g,
40% yield) as a yellow solid after evaporation of the eluant which was taken
forward
directly. RT = 2.37 min (Cond. 3); LC/MS: Anal. Calcd for C23H32N302Si [M+H]+
410.23; found: 410.12.
D-3d2 to D-3d4 were prepared according to the procedures described for D-
3d1.
sI RT = 2.34 min (Cond.
D-3d2 N 3);
LC/MS: Anal. Calcd
for C24H32N302Si
[M+H]+ 422.22; found:
421.85.
From M3, step .\h4
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I
Si
RT = 2.43 min (Cond.
D-3d3 SS,N3);
LC/MS: Anal. Calcd
for C26H34N302Si
[M+H]+ 448.24; found:
448.82.
From D-3c2
Si RT = 2.51 min (Cond.
00 3);
LC/MS: Anal. Calcd
D-3d4
for C26H32N302Si
(M+H)+ 446.23; found:
446.05.
From D-3c3
Example D-3e1 (step e)
i3
Potassium carbonate (91 mg, 0.66 mOmc-oN1))was added in one portion to a
stirred
solution of D-3d1 (2.7 g, 6.6 mmol) in Me0H (60 mL) at rt. The mixture was
stirred
for 1 h before it was concentrated down in vacuo. The residue was taken up in
dichloromethane and added directly to a 130 g Thompson silica gel column.
Gradient elution of the residue with 15% ethyl acetate in hexanes to 100%
ethyl
acetate furnished D-3e1 (2.04 g, 87% yield) as a yellow foam after evaporation
of the
eluant. A small amount (approx. 20 mg) of the product was then subjected to
preparative HPLC to afford a purer sample of D-3e1 as an off-white solid. 1H
NMR
(500 MHz, Me0D, imidazole N-H missing) 6 ppm 7.68 (2 H, d, J=7.9 Hz), 7.46 (2
H, d, J=8.2 Hz), 7.34 - 7.40 (1 H, m), 4.89 (1 H, m), 3.64 - 3.73 (1 H, m),
3.49 (1 H,
m), 3.49(1 H, s), 3.31 - 3.34 (1 H, m), 2.25 - 2.44 (1 H, m), 1.99 - 2.11 (3
H, m), 1.25
and 1.47 (9 H, 2s). RT = 1.63 min (Cond. 3), LC/MS: Anal. Calcd for C20H24N302
(M+H)+ 338.19; found: 338.12.
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D-3e2 to D-3e5 were prepared according to the procedures described for
alkyne D-3e 1.
RT = 1.65 mm (Cond. 3);
D-3e2 HN/ LC/MS: Anal. Calcd for
Boc-N\4
C21H24N302 [M+1-1]+
350.19; found: 350.10.
From D-3d2
010 RT =
1.81 mm (Cond. 3);
D-3e3 LC/MS: Anal. Calcd for
Boc-Nx)
C23H26N302 [M+1-1]+
376.20; found: 376.20.
From D-3d3
RT = 1.88 mm (Cond. 3);
D-3e4 LC/MS: Anal. Calcd for
Boc-Ni)
- C23H24N302 (M+H)+
374.19; found: 374.04.
From D-3d4
N
Wo "Nµ\
N_kbõ
0
RT = 1.75 mm (Cond. 3);
D-3e5 ¨c) N
LC/MS: Anal. Calcd for
51 using the procedures 431.21; found: 431.09.
outlined for D-3h1 and
Example OL-1
Example D-3f1 (step f)
/1\1,Boc
\\µ'
,
Boc
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Tetrakis(triphenylphosphine)palladium (83 mg, 0.072 mmol) was added in
one portion to a stirred, argon-degassed mixture of Example D-3c1 (300 mg,
0.717
mmol), D-3e1 (315 mg, 0.932 mmol), triethylamine (0.40 mL, 2.87 mmol) and
copper(I) iodide (13.7 mg, 0.072 mmol) in anhydrous DMF (6 mL) at rt in a
thick-
walled, screw-top vial. The mixture was stirred at rt for 16 h and at 40 C
for 16 h
before additional CuI (10 mg), TEA (0.4 mL) and Pd(PPft3)4 catalyst (40 mg)
were
added since the reaction was not complete after 32 h as judged by LCMS. The
mixture was stirred further at 60 C for 10 h before it was cooled to rt,
diluted with
ethyl acetate, THF and water and suction-filtered. The organic phase of the
filtrate
was separated, washed with saturated sodium bicarbonate solution and brine
prior to
drying over anhydrous sodium sulfate and evaporation. The residue was taken up
in
dichloromethane and added directly to a Thompson 80 g silica gel column.
Gradient
elution of the residue with 30% ethyl acetate in hexanes to 100% ethyl acetate
followed by 0% methanol in ethyl acetate to 20% methanol in ethyl acetate
furnished
D-3f1 (366.7 mg, 68% yield) as a golden brown foam after evaporation of the
eluant.
A small amount (approx. 20 mg) of product was then subjected to preparative
HPLC
to afford a purer sample of Example D-3f1 as a tan solid. 1H NMR (500 MHz,
Me0D, imidazole N-H's missing) 6 ppm 7.73 (2 H, d, J=8.2 Hz), 7.47 - 7.57 (4
H,
m), 7.41 (2 H, br s), 4.96 - 5.09 (1 H, m), 4.89 - 4.96 (1 H, m), 3.69 (2 H,
br s), 3.54
(2 H, br s), 3.04 - 3.13 (2 H, m), 2.83 - 2.94 (2 H, m), 2.30 -2.50 (2 H, m),
1.94 -
2.17 (6 H, m), 1.27 - 1.49 (18 H, 2s). RT = 2.06 min (Cond. 3); LC/MS: Anal.
Calcd
for C40H47N604 (M+H)+ 675.37; found: 675.26.
D-3f2 to D-3f4 were prepared according to the procedure described for D-3f1.
RT = 2.12 min
Boc
H (Cond. 3); LC/MS:
Anal. Calcd for
D-3f2
Boc'
C42H45N604 [M+H]+
697.35; found:
D-3e2 and D-3c3
697.25.
D-3f3 RT = 2.14 min
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I
,Boc (Cond. 3); LC/MS: TC r
¨ / N Anal. Calcd for
H
v , r
4449,1-NT 6vn
4 j+
Boc'
725.38; found:
From D-3e3 and D-3c2 725.25.
Boc
H
LCMS: 2.18 min
¨ N
\ \\
M (Cond. 3); LC/MS:
D-3f4
Anal. Calcd for
C45H48N705 (M+H)+
778.37; found:
From D-3c3 and D-3e5
778.22.
Example D-3g1 (step g)
,Boc
N
Nr11)--
N = ¨ N \
H N
Boc'
Activated manganese dioxide (2.0 g, 23.12 mmol) was added in one portion
to a stirred solution of D-3f1 (260 mg, 0.39 mmol) in dry dichloromethane (5
mL).
The suspension was stirred at rt for 6 h before additional activated manganese
dioxide (1.0 g) was added. The suspension was stirred further for 14 h at rt
before it
suction-filtered through a pad of Celite with Me0H and the pad was washed with
additional Me0H several times. The filtrate was then concentrated in vacuo to
yield
D-3g1 (225.1 mg, 82% yield) as a yellow solid. A small amount (approx. 20 mg)
of
the product was then subjected to preparative HPLC to afford a purer sample of
D-
3g1 as a light tan solid. 1H NMR (500 MHz, Me0D, imidazole N-H's missing) 6
ppm 8.43 (1 H, d, J=8.5 Hz), 8.19 (1 H, br s), 7.68 - 7.78 (5 H, m), 7.62 (2
H, d,
J=8.2 Hz), 7.54 (1 H, s), 4.90 - 5.27 (2 H, m), 3.76 - 3.85 (1 H, m), 3.66 -
3.74 (1 H,
m), 3.51 - 3.66 (2 H, m), 2.35 -2.60 (2 H, m), 1.94 - 2.21 (6 H, m), 1.49 (6
H, br s),
1.27 (6 H, br s), 1.14 (6 H, br s). RT = 2.12 min (Cond. 3); LC/MS: Anal.
Calcd for
C40H45N604 (M+H)+ 673.35; found: 673.30.
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D-3g2 was prepared according to the procedure described for D-3g1.
.Boc LCMS:
2.27 min
N (Cond. 3); LC/MS:
N=`"
D-3g2 H Anal.
Calcd for
Boc
C44H45N604 (M+H)+
From D-3f3 721.35; found:
721.25.
Example D-3h1 (step h)
HND
D-3h1 to D-3h6 were prepared from D-3f1, D-3g1, D-3f2, D-3f3, D-3g2 and
D-3 f4, respectively, according to the same procedure used for the preparation
of OL-
le except that methanol (1 mL) was used instead of dichloromethane. This gave
D-
3h1 to D-3h6 as hydrochloride salts (or TFA salts when purified further with
preparative HPLC) upon concentration of the solvent(s) in vacuo.
RT = 1.44 min (Cond. 3);
D-3h1
µµ,NcENi
LC/MS: Anal. Calcd for
C30F131N6 [M+H]+ 475.26;
found: 475.16.
From D-3f1
1H NMR (500 MHz, Me0D,
imidazole N-H's missing) 6
ppm 8.46 (1 H, d, J=8.5
D-3h2
Hz), 8.21 (1 H, s), 7.84 (2
\\µµ IN\ // ¨1/0
N µ`µ
H H, d,
J=8.2 Hz), 7.70 - 7.80
From D-3g1 (4 H,
m), 7.64 (2 H, d, J=8.5
Hz), 5.15 (1 H, s), 4.97 (1
H, s), 3.62 - 3.70 (1 H, m),
3.56 (3 H, s), 2.65 - 2.75 (1
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H, m), 2.55 - 2.64 (1 H, m),
2.17 - 2.53 (6 H, series of
m). RT = 1.61 min (Cond.
3); LC/MS: Anal. Calcd for
C301-129N6 (M+H)+ 473.25;
found: 473.13.
'NHH RT = 1.62 min (Cond. 3);
D-3h3
¨ N LC/MS: Anal. Calcd for
N = ¨ = = LC/MS: Anal.
(M+H)+ 497.25;
From D-3f2 found: 497.13.
RT = 1.62 min (Cond. 3);
NH
D-3h4'H LC/MS: Anal. Calcd for
\µµ ¨ N
N ¨ C34H33N6 [M+H]+ 525.28;
found: 525.13.
From D-3f3
'NH
H LCMS: 1.78 min (Cond. 3);
D-3h5 ¨ N
N_N LC/MS: Anal. Calcd for
HN-.4ek C34H29N6 (M+H)+ 521.25;
From D-3g2 found: 521.13.
NH
LCMS: 1.95 min (Cond. 3);
D-3h6 LC/MS: Anal. Calcd for
H
0
C41H40N703 (M+H)+
678.32; found: 678.45.
From D-3f4
Example D-3 to D-11 (final step)
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N 0
[ H ____________________________
CM1121
N _______________________________ - = = N'-'1,=\)
0,1\1
H
Examples D-3 to D-11 were prepared from D-3h1 to D-3h6 and the
appropriate acids according to the same procedure used for the preparation of
Example OL-1. This gave Examples D-3 to D-11 as TFA salts after HPLC
purification.
RT = 1.80 min (Cond. 3);
LC/MS: Anal. Calcd for
C44H53N806 [M+1-1]+
789.41; found: 789.27. 1H
NMR (500 MHz, Me0D,
imidazole N-H's missing) 6
ppm 7.86 - 7.91 (1 H, m),
7.73 - 7.80 (2 H, m), 7.64
7.71 (2 H, m), 7.50 - 7.59
r-N 0
D-3
(3 H, m), 5.59 - 5.65 and
\""Y3 -
N = - = ' N),=1.) 5.16 - 5.31 (2 H, 2m), 4.21
(7).1\1
- 4.29 (2 H, m), 4.06 - 4.16
N
H (2 H, m), 3.81 - 3.93 (2 H,
From D-3h1 and Cap 51 m), 3.67 (6 H, s), 3.15 -
3.24 (2 H, m), 2.95 - 3.07
(2 H, m), 2.51 -2.65 (2 H,
m), 2.25 - 2.37 (2 H, m),
2.14 -2.23 (4 H, m), 1.99 -
2.11 (2 H, m), 0.09 -0.95
(12 H, m), 0.97 - 1.05 (2
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H, m).
0111 11\11-1
RT = 1.88 min (Cond. 3);
D-4
N LC/MS: Anal. Calcd for
/Nj \ C50H49N806 [M+1-1]+
H N
o 0 857.38; found: 857.31.
¨o.Lri
From D-3h1 and Cap 4
4111NH
RT = 1.94 mm (Cond. 3);
D-5
\\, LC/MS: Anal. Calcd for
N C50H47N806 [M+1-1]+
H N
o 0 855.36; found: 855.28.
From D-3h2 and Cap 4
0 0
),INH
RT = 1.88 mm (Cond. 3);
cy
D-6 LC/MS: Anal. Calcd for
N_N
Ns¨> C44H51N806 [M+1-1]+
787.40; found: 787.30.
From D-3h2 and Cap 51
0 0 ===.,
NLO
RT = 1.93 mm (Cond. 3);
D-7 \\"Y2)I, ¨ \¨/ N LC/MS: Anal. Calcd for
N ' ¨ ' 1\13µ`\\
C46H51N806 [M+1-1]+
811.40; found: 811.26.
From D-3h3 and Cap 51
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= NH
Nr_., 2 RT = 2.01 min (Cond. 3);
D-8 \\" 1 N, // % /¨--/ .,1\1 ,\ =--N LC/MS: Anal. Calcd for
= N----/..k C52H47N806
[M+1-1]+
o o
---oLN . 879.36; found: 879.25.
H
From D-3h3 and Cap 4
ZNH
RT = 1.93 mm (Cond. 3);
D-9 = 2
LC/MS: Anal. Calcd for
N / c48H55N806 [M+1-1]+
N
o 839.43; found: 839.26.
5-- ..---f
¨o
From D-3h4 and Cap 51
0,,,o,
)..õZNH
N 0 RT = 2.07 mm (Cond. 3);
D-10 \µ'il5 ¨ / N LC/MS: Anal. Calcd for
N-- \
H 14,
C48H51N806 [M+1-1]+
o N__)
5-- .----\/ 835.40; found: 835.26.
---o N
H
From D-3h5 and Cap 51
00.,,,2/H
N 0 RT = 1.99 mm (Cond. 3);
D-11 Yid ¨ / N LC/MS: Anal. Calcd for
N-- \
H N__>lik
O C50H53N807 [M+1-1]+
877.41; found: 877.63.
¨o
From D-3h6 and Cap 170
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Example M1
Me02CHN
N--,./-"N ¨ 0 --:-.-
..---- NHCO2Me
Example Ml, Step a
n
Boc )_(¨)__<.,11-1\1%
¨
Pd(Ph3P)4 (0.078 g, 0.067 mmol) was added to a DMF (5 mL) solution of (5)-
tert-butyl 2-(5-(4-bromopheny1)-1H-imidazol-2-y1)pyrrolidine-1-carboxylate(D-
lb)
(0.675g, 1.721 mmol) and 1,2-bis(trimethylstannyl)ethyne (0.3134 g, 0.891
mmol) in
a pressure tube, the mixture was flushed with nitrogen for 1 min and then
heated at
90 C for 15 h. The volatile component was removed in vacuo and the residue
was
directly submitted to a BIOTAGEO purification (110 g; Et0Ac) to afford alkyne
Mla as a yellowish orange foam containing unidentified impurity and residual
solvent (430 mg). 1H NMR (DMSO, 6 = 2.50 ppm, 400 MHz): 12.23/11.98/11.91
(three br s, 2H), 7.80-7.35 (m, 10H), 4.84-4.76 (m, 2H), 3.54 (m, 2H), 3.39-
3.33 (m,
2H), 2.28-1.78 (m, 8H), 1.40 (br s, 7.54H), 1.16 (br s, 10.46H). LC/MS: Anal.
Calcd.
for [M+H]+ C381445N604: 649.35; found 649.27.
Example Ml, Step b
-7----)
U
HC1/dioxanes (4N; 8 mL, 32.0 mmol), CH2C12 (1 mL) and Me0H (1.0 mL)
were added to bis-carbamate Mla (0.427 g, 0.658 mmol), and the heterogeneous
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mixture was stirred for 5 hr. The volatile component was removed in vacuo, and
the
residue was exposed to high vacuum to afford Mlb (.4HC1) as an off-white solid
(452 mg), which was submitted to the next step as such. 1H NMR analysis
indicates
that the sample may contain residual dioxane (-1 mol equiv). 1H NMR (DMSO, 6 =
2.50 ppm, 400 MHz): 10.29 (br s, 2H), 9.73 (br s, 2H), 8.11 (s, 2H), 7.96 (d,
J = 8.3,
4H), 7.67 (d, J = 8.6, 4H), 4.97 (br m, 2H), 3.47-3.31 (m, 4H), 2.50-2.36 ('m'
partially overlapped with solvent signal, 4H), 2.23-2.14 (m, 2H), 2.07-1.95
(m, 2H).
LC/MS: Anal. Calcd. for [M+H]+ C28F129N6: 449.25; found 49.23.
Example M1
HATU (0.069 g, 0.181 mmol) was added to the DMF solution of pyrrolodine
Mlb/4HC1 (70.5 mg, 0.103 mmol), (S)-2-(methoxycarbonylamino)-3-methylbutanoic
acid (36.3 mg, 0.207 mmol) and DIEA (0.1 mL, 0.573 mmol) and stirred at room
temperature for 70 min. The volatile component was removed in vacuo and the
residue was dissolved in Me0H and submitted to a reverse phase HPLC
purification
(Me0H/water/TFA) to afford the TFA salt of Example M1 as an off-white foam
(68.9 mg). LC (Cond. 9 and 10): >95% homogeneity index. LC/MS (Cond. 3): Rt =
1.78 min. 1H NMR (DMSO, 6 = 2.50 ppm, 400 MHz): 8.10 (br s, 2H), 7.88-7.83 (m,
4H), 7.71 (d, J = 8.3, 3.91H), 7.34 (d, J = 8.5, 2H; NHCO2), 6.92 (app br s,
0.09H),
5.52 (br m, 0.17H), 5.12 (app t, 1.93H), 4.11 (app t, 2H), 3.89-3.77 (m, 4H),
3.54 (s,
5.52H), 3.33 (s, 0.48H), 2.41-2.33 (m, 2H), 2.21-1.93 (m, 8H), 0.89 (app t,
0.91H),
0.83/0.79 (two overlapping d, J = 6.8/6.8, 11.09 H). LC/MS: Anal. Calcd. for
[M+H]+ C42H51N806: 763.39; found 763.33.
Example M2 to M2.1
Example M2 to M2.1 were prepared as TFA salts from pyrrolidine Mlb and
appropriate acids according to the procedure described for the preparation of
Example M1 with a modified purification protocol noted in the table below.
i--
NH R
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Example R Analytical data
M2,IS HCO2Me
µPh Purified using two different reverse phase
HPLC conditions (Column: Phenomenex Luna,
0 N
30x100 mm, S10; Me0H/water/TFA) and
(Waters Sunfire, 30x100 mm, S5;
CH3CN/water/TFA). LC (Cond. 9 and 10):
>95% homogeneity index. LC/MS (Cond. 3):
Rt = 2.04 min. LC/MS: Anal. Calcd. for
[M+H]+ C48H47N806: 831.36; found 841.41.
M2.1 (0, Purified using two different reverse phase
HPLC conditions (Column: Xbridge, 19x100
JS, r mm, S5; Me0H/water/TFA) and (Waters
0 NHCO2Me
Sunfire, 30x100 mm, S5; CH3CN/water/TFA).
LC (Cond. 9 and 10): 95% homogeneity index.
LC/MS (Cond. 10d): Rt = 2.82 min. LC/MS:
Anal. Calcd. for [M+H]+ C46H55N808: 847.41;
found 847.25.
Example M3
Me02CHN
\ NH
0 %
4---4 NHCO2Me
Example M3, Step a
TBDPS¨C)x\µ,.
0
N
H
To a solution of (S)-5-(hydroxymethyl)pyrrolidin-2-one (10 g, 87 mmol) in
CH2C12 (50 mL) was added tert-butylchlorodiphenylsilane (25.6 g, 93 mmol),
Et3N
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(12.1 mL, 87 mmol) and DMAP (1.06 g, 8.7 mmol). The mixture was stirred at
room
temperature until the starting pyrrolidinone was completely consumed, and then
it
was diluted with CH2C12 (50 mL) and washed with water (50 mL). The organic
layer
was dried (Na2SO4), filtered, and evaporated in vacuo, and the crude material
was
submitted to flash chromatography (silica gel; 30 to 100% of Et0Ac/hexanes) to
afford ether M3a as a colorless oil (22.7 g, 74% yield). 1H-NMR (400 MHz, DMSO-
d6, 6 = 2.5 ppm) 7.69 (br s, 1H), 7.64-7.61 (m, 4H), 7.50-7.42 (m, 6H), 3.67-
3.62 (m,
1H), 3.58-3.51 (m, 2H), 2.24-2.04 (m, 3H), 1.87-1.81 (m, 1H), 1.00 (s, 9H).
LC/MS
(M+H)+ = 354.58.
Example M3, Step b
TBDPS' \\\,= o
N
I
Boc
Di-tert-butyl dicarbonate (38.5 g, 177 mmol) was added in portions as a solid
over 10 min to a CH2C12 (200 mL) solution of silyl ether M3a (31.2 g, 88.3
mmol),
Example M3, Step c
TBDPS' \\\,=
N
I
Boc
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A three-necked flask equipped with a thermometer and a nitrogen inlet was
charged with carbamate M3b (10.05 g, 22.16 mmol) and toluene (36 mL), and
lowered into -55 C cooling bath. When the internal temperature of the mixture
reached -50 C, lithium triethylborohydride (23 mL of 1.0 M/THF, 23.00 mmol)
was
added dropwise over 30 min and the mixture stirred for 35 min while
maintaining the
internal temperature between -50 C and -45 C. Hunig's base (16.5 mL, 94
mmol)
was added dropwise over 10 min. Then, DMAP (34 mg, 0.278 mmol) was added in
one batch, followed by the addition of trifluoroacetic anhydride (3.6 mL, 25.5
mmol)
over 15 min, while maintaining the internal temperature between -50 C and -45
C.
Example M3, Step d
TBDPS"- \\µµ.C7
N
I
Boc
M3d-1: trans-isomer
M3d-2: cis-isomer
Diethylzinc (19 mL of -1.1 M in toluene, 20.9 mmol) was added dropwise
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was opened to the air and quenched by the slow addition of 50% saturated
NaHCO3
solution (40 mL), and then removed from the cooling bath and stirred at
ambient
temperature for 20 min. It was filtered through a filter paper and the white
cake was
washed with 50 mL of toluene. The organic phase of the filtrate was separated
and
washed with water (40 mL, 2x), dried (MgSO4) and concentrated in vacuo. The
crude material was purified using a BIOTAGEO system (350 g silica gel; sample
was
loaded with 7% Et0Ac/hexanes; eluted with 7-20% Et0Ac/hexanes) to afford a
mixture of methanopyrrolidines M3d-1 and M3d-2 as a colorless viscous oil
(3.69 g,
90.7%). [Note: the exact cis/trans-isomer ratio was not determined at this
stage].
LC/MS (Cond. 7): Rt = 2.39 min 1H-NMR (400 MHz, DMSO-d6, 6 = 2.5 ppm) 7.62-
7.60 (m, 4H), 7.49-7.40 (m, 6H), 3.77/3.67 (overlapping br s, 3H), 3.11-3.07
(m, 1H),
2.23 (app br s, 1H), 2.05-2.00 (m, 1H), 1.56-1.50 (m, 1H), 1.33 (very broad s,
9H),
1.00 (s, 9H), 0.80 (m, 1H), 0.30 (m, 1H). LC/MS: [M+Na]+ = 474.14.
Example M3, Step e
HOµ Cy-,\\%.
N
I
Boc
M3e-1: trans-isomer
M3e-2: cis-isomer
TBAF (7.27 mL of 1.0 M in THF, 7.27 mmol) was added dropwise over 5
min to a THF (30 mL) solution of silyl ether M3d-1/-2 (3.13 g, 6.93 mmol) and
the
mixture stirred at ambient temperature for 4.75 h. After the addition of
saturated
NH4C1 solution (5 mL), most of the volatile material was removed in vacuo and
the
residue partitioned between CH2C12 (70 mL) and 50% saturated NH4C1 solution
(30
mL). The aqueous phase was extracted with CH2C12 (30 mL), and the combined
organic phase was dried (Mg504), filtered, concentrated in vacuo and then
exposed
to high vacuum overnight. The crude material was purified using a BIOTAGEO
(silica gel; 40-50% Et0Ac/hexanes) to afford a mixture of alcohols M3e-1 and
M3e-
2, contaminated with traces of a lower Rf spot, as a colorless oil (1.39 g,
¨94% yield).
[Note: the exact cis/trans isomer ratio was not determined at this stage.] 1H-
NMR
(400 MHz, DMSO-d6, 6 = 2.5 ppm) 4.70 (t, J = 5.7, 1H), 3.62-3.56 (m, 1H), 3.49-
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3.44 (m, 1H), 3.33-3.27 (m, 1H), 3.08-3.04 (m, 1H), 2.07 (br m, 1H), 1.93-1.87
(m,
1H), 1.51-1.44 (m, 1H), 1.40 (s, 9H), 0.76-0.71 (m, 1H), 0.26 (m, 1H). LC/MS
(M+Na)+ = 236.20.
Example M3, Step f
HO C7
0 I
Boc
M3f-1: trans isomer
M3f-2: cis isomer
A semi-solution of NaI04 (6.46 g, 30.2 mmol) in H20 (31 mL) was added to a
solution of alcohol M3e-1/-2 (2.15 g, 10.08 mmol) in CH3CN (20 mL) and CC14
(20
mL). RuC13 (0.044 g, 0.212 mmol) was added immediately and the heterogeneous
reaction mixture was stirred vigorously for 75 min. The reaction mixture was
diluted
with H20 (60 mL) and extracted with CH2C12 (50 mL, 3x). The combined organic
phase was treated with 1 mL Me0H, allowed to stand for about 5 min, and then
filtered through a pad of diatomaceous earth (CELITEO). The CELITEO was
washed with CH2C12 (50 mL), and the filtrate was concentrated in vacuo to
afford a
light charcoal-colored solid. 1H-NMR analysis of this crude material indicated
a
1.00:0.04:0.18 mole ratio of trans acid M3f-1:cis acid M3f-2:presumed side
product,
tert-butyl 3-oxo-2-azabicyclo[3.1.0]hexane-2-carboxylate. The crude material
was
dissolved in Et0Ac (-10 mL) with heating and allowed to stand at ambient
temperature with seeding. About 15 min into the cooling phase, a rapid crystal
formation was observed. About 1 h later, hexanes (-6 mL) was added and the
mixture refrigerated overnight (it did not appear that additional material
precipitated
out). The mixture was filtered and washed with ice/water-cooled hexanes/Et0Ac
(2:1 ratio; 20 mL) and dried under high vacuum to afford the first crop of
acid M3f-1
(off-white crystals, 1.222 g). The mother liquor was concentrated in vacuo,
and the
residue dissolved in ¨3 mL of Et0Ac with heating, allowed to stand at ambient
temperature for 1 h, and then 3 mL hexanes was added and stored in a
refrigerator for
¨15 h. A second crop of acid M3f-1 was retrieved similarly (grey crystals,
0.133 g),
for a combined yield of 59%. Acid M3f-1: Rt = 1.48 min under the following
HPLC
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conditions: Solvent gradient from 100% A : 0% B to 0% A: 100% B (A = 0.1% TFA
in 1:9 Me0H/H20; B = 0.1% TFA in 9:1 Me0H/H20) over 3 min; detection @220
nm; PHENOMENEXO-Luna 3.0X50 mm S10 column. MP (dec.) for the first crop =
147.5-149.5 C. 1H-NMR (400 MHz, DMSO-d6, 6 = 2.5 ppm) 12.46 (s, 1H), 3.88
(app br s, 1H), 3.27 (app br s, 1H; overlapped with water signal), 2.28 (br m,
1H),
2.07 (app br s, 1H), 1.56 (app s, 1H), 1.40/1.34 (two overlapped s, 9H), 0.71
(m, 1H),
0.45 (m, 1H). 13C-NMR (100.6 MHz, DMSO-d6, 6 = 39.21 ppm) 172.96, 172.60,
154.45, 153.68, 78.74, 59.88, 59.58, 36.91, 31.97, 31.17, 27.77, 27.52, 14.86,
14.53,
13.69. LC/MS [M+Na]+ = 250.22. Anal. Calcd. for C11H12N04: C, 58.13; H, 7.54;
N, 6.16. Found (for first crop): C, 58.24; H, 7.84; N, 6.07. Optical rotation
(10
mg/mL in CHC13): [a]r) = -216 and -212 for the first and second crop,
respectively.
Example M3, Step g
C),) ,--.3>
Br 0 .
0
/
Boc
DIEA (1.3 mL, 7.44 mmol) was added dropwise over 2 min to a semi-
heterogeneous mixture of acid M3f-1 (1.697 g, 7.47 mmol) and 2-bromo-1-(4-
bromophenyl)ethanone (2.01 g, 7.23 mmol) in CH3CN (30 mL) and stirred at room
temperature for 8 hr. The volatile component was removed in vacuo and the
residue
was taken up in CH2C12 (100 mL), washed with water (30 mL), dried (Mg504) and
concentrated in vacuo to afford ketoester M3g as an off-white viscous semi-
foamy oil
(3.076 g). 1H-NMR (400 MHz, DMSO-d6, 6 = 2.5 ppm): 7.92 (d, J = 8.3, 2H), 7.78
(d, J = 8.5, 2H), 5.61-5.42 (m, 2H), 4.16 (m, 1H), 3.34 ('m' partially
overlapped with
water signal, 1H), 2.40 (m, 2H), 1.63 (m, 1H), 1.41/1.35 (two overlapped 's',
9H),
0.74 (m, 1H), 0.53 (m, 1H). LC/MS: Anal. Calcd. for [M+Na]+ C19H2279BrNNa05:
446.06; found 446.06.
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Example M3, Step h
H7
Br 4. N-...7.¨N
\ Boo
A 350 ml pressure tube was charged with keto-ester M3g (3.07g, 7.24 mmol),
ammonium acetate (5.48 g, 71.1 mmol) and Xylene (70 mL), capped and heated
with
an oil bath (140 C) for 4.5 hr. The reaction mixture was allowed to cool to
room
temperature and the volatile component was removed in vacuo. CH2C12 (100 mL)
and
50% saturated NaHCO3 solution (30 mL) were added to the residue, vigorously
stirred until gas evolution ceased, and the phases were separated. The organic
layer
was dried (Mg504), concentrated in vacuo, and purified with a BIOTAGEO (240 g
silica gel; sample was loaded with CH2C12; 40-80% Et0Ac/hexanes) to afford
imidazole M3h as a dark yellow foam (2.40 g). 1H-NMR (400 MHz, DMSO-d6, 6 =
2.5 ppm): 12.21 (s, 0.11H), 1.93 (s, 0.89H), 7.69 (d, J = 8.8, 1.8H), 7.62-
7.55 (m,
0.4H), 7.53 (br d, J = 2, 0.87H), 7.49 (d, J = 8.5, 1.8H), 7.29 (br d, J =
1.6, 0.13H),
4.59 (m, 1H), 3.41 (m, 1H), 2.37-2.17 (br m, 2H), 1.62 (m, 1H), 1.21 (very
broad 's',
9H), 0.75 (m, 1H), 0.54 (m, 1H). LC/MS: Anal. Calcd. for [M+H]+ C16I-
12381BrN302:
406.10; found, 406.14.
Example M3, Step i
zt---7
Pyrrolidine M3i (4HC1) was prepared from bromide M3h according to the
procedure described for the synthesis of pyrrolidine M lb (.4HC1) from bromide
D-
lb. 1H-NMR (400 MHz, DMSO-d6, 6 = 2.5 ppm): 10.51 (app br s, 4H), 8.08 (s,
2H),
7.92 (d, J= 7.8, 4H), 7.66 (d, J = 8.5, 4H), 4.78 (m, 2H), 3.42 (m, 2H), 2.65
(m, 2H),
¨2.53 ('m' partially overlapped with solvent signal, 2H), 1.94 (m, 2H), 1.10
(m, 2H),
0.86 (m, 2H). LC/MS: Anal. Calcd. for [M+H]+ C30H29N6: 473.25; found 473.21.
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Example M3 (and Examples M4-M7)
Example M3, along with its analogs Examples M4-M7 highlighted in the
table below, were prepared as TFA salts from pyrrolidine M3i (.4HC1) by
employing
the procedure described for the synthesis of Example M1 and appropriate acids.
In
the case of Example M7 an equimolar mixture of (S)-2-(methoxycarbonylamino)-3-
methylbutanoic acid and (S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-
yl)acetic acid was employed for the coupling step and the resultant
statistical mixture
of products was separated by the HPLC technique described for Example Ml.
Example M3: LC (Cond. 9 and 10): >95% homogeneity index. LC/MS (Cond. 3): Rt
= 1.89 min. LC/MS: Anal. Calcd. for [M+H]+ C44H511\1806: 787.39; found 787.40.
H C-57
R--o NO__ _____________________ ) (_¨)__ CV'. 0 IR NI\
__________________________________ ¨ \ ___ / \ N ---
A163 H
Example
IR--0 IR--o Rt (Cond. 3); % homogeneity
index (Cond. 9 and 10); LC/MS
data
M4 o/ o/ 1.74 min; >95%; LC/MS: Anal.
Calcd. for [M+I-1]+ C42H47N806:
FINHN
0
2....,..e 759.36; found 759.35
(.... \ \J=Pri
M5 o/ o/ 1.98 min; >95%; LC/MS: Anal.
HN/0
HNO Calcd. for [M+H]+ C46H51N806:
: 0
811.39; found 811.37
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Example Rt (Cond. 3); % homogeneity
index (Cond. 9 and 10); LC/MS
Jsoss
data
M6 o/ o/ 1.67 min; >95%; LC/MS: Anal.
HNO
HNO Calcd. for [M+H]+ C48H551\1808:
0
871.41; found 871.20
rPri "Pi
0 0
M7 o/ o/ 1.78 min; >95%; LC/MS: Anal.
HNO
HNO Calcd. for [M+H]P C46H53N807:
829.40; found 829.53
rrrj Prrj
0
Examples M8-M9
Me02CHN 0
HN
\ NH
N
Example M8 (R = H) CI ¨NHCO2Me
Example M9 (R = CI)
NCS (0.0195 g, 0.143 mmol) was added to a DMF (2 mL) solution of the free
base form of Example M3 (obtained from the TFA salt via a standard MCX free-
basing protocol; 0.109 g, 0.139 mmol), and stirred at room temperature for 16
hr and
at 50 C for 25 hr. Most of the solvent was removed in vacuo, and the residue
was
dissolved in Me0H and submitted to a reverse phase HPLC purification
(Me0H/water/TFA) to afford the TFA salt of Example M8 (50 mg) and Example M9
(17.5 mg).
Example Rt (Cond. 3); % homogeneity index (Cond. 9a and 10a);
LC/MS data
M8 2.42 min; >95%; LC/MS: Anal. Calcd. for [M+H]+ C44H50C11\1806:
821.35; found 821.31
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Example Rt (Cond. 3); % homogeneity index (Cond. 9a and 10a);
LC/MS data
M9 2.95 min; >95%; LC/MS: Anal. Calcd. for [M+H]+ C44H49C121\1806:
855.32; found 855.24
Example M9.1-M9.2
0
Me02CHN
u
h,¨NH n =
Example M9.1 (R= H) CI --NHCO2Me
Example M9.2 (R = CI)
NCS (0.021 g, 0.158 mmol) was added to a DMF (1.5 mL) solution of the
free base form of Example M6 (obtained from the TFA salt via a standard MCX
free-
basing protocol; 0.1059 g, 0.122 mmol), and stirred at 50 C for 24 hr. The
reaction
mixture was diluted with Me0H (2.5 mL) and submitted to a reverse phase HPLC
purification condition (XTERRA, 30X100 mm, S5; Me0H/water/TFA). The
resultant sample was repurified with a different reverse phase HPLC condition
(Waters-Sunfire, 30X100 mm, S5; acetonitrile/water/TFA) to afford Example M9.1
(38.8 mg) and Example M9.2 (32.6 mg) as TFA salts.
Example Rt (Cond. 3); % homogeneity index (Cond. 9a and 10a);
LC/MS data
M9.1 2.26 min; >95%; LC/MS: Anal. Calcd. for [M+H]+ C48F154C1N808:
905.38; found 905.44
M9.2 2.78 min; >95%; LC/MS: Anal. Calcd. for [M+H]+ C44H53C121\1808:
939.34; found 939.40
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Example M9.3
¨0 H
NH0 \ II
H 0--
Example M9.3, step a
41, A =
To a cooled (0 C) CH2C12 (22 mL) was added diethylzinc (1M in hexanes,
60.5 mL, 60.5 mmol) under nitrogen followed by the dropwise addition of
trifluoroacetic acid (5.1 mL, 66.2 mmol) in 10 mL CH2C12 over 15 min. The
reaction was stirred for 15 min, and then diiodomethane (5.4 mL, 66.9 mmol) in
10
mL CH2C12 was added dropwise to the reaction. The reaction was continued to
stir
at 0 C for 1 hr, and then (E)-1,2-diphenylethene (2 g, 11.10 mmol) in 10 mL
CH2C12 was added. The mixture was removed from the cold bath and stirred at
¨25 C under nitrogen for 20 h. The reaction was quenched with 0.1N HC1 (50
mL),
the layers were separated, and the aqueous layer was extracted with hexanes (2
x 100
mL). The combined organic phase was washed with saturated NaHCO3 (aq) (50
mL) and brine (50 mL), dried over MgSO4, filtered, and concentrated in vacuo.
The
residue was submitted to a silica gel flash chromatography (hexanes) to afford
biphenyl M9.3a as a colorless oil (1.6 g). 1H NMR (400 MHz, CDC13) 6 ppm 7.34-
7.31 (m, 4H), 7.24-7.17 (m, 6H), 2.22-2.18 (m, 2H), 1.51-1.47 (m, 2H).
Example M9.3, step b
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0 Br 0 Br
+
0 lei
Br Br
M9.3b1 M9.3b2
To a solution of biphenyl M9.3a (3.3 g, 16.99 mmol) in 1,2-dimethoxyethane
(197 mL) and water (82 mL) was added NBS (12.09 g, 67.9 mmol). The reaction
flask was covered with aluminum foil and stirred at room temperature for 51
hr. The
reaction was partitioned between diethyl ether and water, and the organic
layer was
washed with brine, dried over MgSO4, filtered, and concentrated in vacuo. The
residue was submitted to a flash chromatography (sample was loaded with
chloroform; eluted with hexanes) to afford a stereoisomeric mixture of
dibrominated
product as a white solid (3.1 g). The stereoisomeric mixture was separated by
chiral
SFC (Chiral pak AD-H column, 30X250 mm, 5 pm; 80% CO2-20% Et0H; 35 C;
150 bar; 70 mL/min for 20 min; 220 nm) to isolate two enantiomers: M9.3b1
(white
solid, 1.1 g); 1H NMR (400 MHz, CDC13) 6 ppm 7.43 (d, J = 8.6 Hz, 4H), 7.02
(d, J
= 8.6 Hz, 4H), 2.12-2.09 (m, 2H), 1.47-1.43 (m, 2H); OR: +361.79, 3.15 mg in 1
mL
CHC13, 2, = 589 nm, 50 mm cell. M9.3b2 (white solid, 1.2 g); 1H NMR (400 MHz,
CDC13) 6 ppm 7.43 (d, J = 8.6 Hz, 4H), 7.02 (d, J = 8.6 Hz, 4H), 2.12-2.09 (m,
2H),
1.47-1.43 (m, 2H); OR: -376.70, 3.03 mg in 1 mL CHC13, 2, = 589 nm, 50 mm
cell.
Example M9.3, Step c
_ 0
0 ¨
To a solution of dibromide M9.3b1 (0.8806 g, 2.501 mmol) and tributy1(1-
ethoxyvinyl)stannane (2.71 g, 7.50 mmol) in 1,4-dioxane (17 mL) was added
dichlorobis(triphenylphosphine)-palladium(II) (0.105 g, 0.150 mmol). The
reaction
was thoroughly flushed with nitrogen, sealed, and heated at 80 C for 16 h.
The
reaction was removed from the heat, 1N HC1 (aq) (17 mL) was added, and the
mixture was stirred for 4 h. The mixture was diluted with water (20 mL) and
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extracted with Et0Ac (3 x 50 mL). The combined organic layer was dried over
MgSO4, filtered, and concentrated in vacuo. The residue was purified with a
flash
chromatography (35% ethyl acetate/hexanes), and the retrieved sample was
triturated
with hexanes (3 x 50 mL) to afford ketone M9.3c as an off-white solid (412.5
mg).
1H NMR (400 MHz, CDC13) 6 ppm 7.92 (d, J = 8.5 Hz, 4H), 7.23 (d, J = 8.6 Hz,
4H),
2.61 (s, 6H), 2.32-2.28 (m, 2H), 1.67-1.63 (m, 2H). LC/MS: Anal. Calcd. for
[M+H]+ C19H1902: 279.14; found 279.13.
Example M9.3, Step d
To a solution of ketone M9.3c (0.4072 g, 1.463 mmol) in THF (7 mL) was
added phenyltrimethylammonium tribromide (1.10 g, 2.93 mmol), and the reaction
mixture was stirred at ¨25 C for 15 h. The volatile component was removed in
vacuo, and the residue was partitioned between water (25 mL) and CH2C12 (100
mL).
The organic layer was dried over Mg504, filtered, and concentrated in vacuo to
afford dibromide M9.3d, which was used without further purification. LC/MS:
Anal.
Calcd. for [M+H]+ C19H1279Br202: 434.96; found 434.98.
Example M9.3, Step e
_ 0
0 \ / Boc oo 0 µ1\1
0 ¨
o)% _________________________________________________ c::'s<
To a solution of dibromide M9.3d (-1.463 mmol) and acid M3f1 (0.698 g,
3.07 mmol) in acetonitrile (10 mL) was added N,N-diisopropylethylamine (0.537
mL, 3.07 mmol), and the reaction was stirred at ¨25 C for 5h. The volatile
component was removed in vacuo, and the residue was taken up in chloroform (4
mL) and loaded onto a silica gel column and eluted with 8% ethyl
acetate/methylene
chloride over 1296 mL solvents to afford diketoester M9.3e as a light yellow
foam
containing unidentified impurities (0.723 g). 1H NMR (400 MHz, DMSO-d6) 6 ppm
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7.90 (d, J = 8.0 Hz, 4H), 7.37 (d, J = 8.3 Hz, 4H), 5.59-5.40 (m, 3H), 4.24-
4.09 (m,
2H), 3.37-3.25 (m, 2H), 2.47-2.36 (m, 5H), 1.87-1.80 (m, 2H), 1.72-1.68 (m,
2H),
1.57-1.49 (m, 2H), 1.41 (s, 7H), 1.35 (s, 11H), 0.80-0.67 (m, 2H), 0.58-0.48
(m, 2H).
LC/MS: Anal. Calcd. for [M+H]+ C41F148 N2 Na Om: 751.32; found 751.55.
Example M9.3, Step f
______________________________________________ H7
¨ N N
II '
Boc
Boc NII \ / \ ""--r N
A mixture of diketoester M9.3e (0.723 g, 0.992 mmol) and ammonium
acetate (1.529 g, 19.84 mmol) in xylene (10 mL) in a sealed reaction vessel
was
heated at 140 C for 2.5 h. After the reaction was allowed to cool to ambient
condition, the volatile component was removed in vacuo. The residue was taken
up
in 20% Me0H/CHC13 (50 mL) and treated with saturated NaHCO3 (aq) (20 mL),
stirred and the layers were separated. The aqueous layer was extracted further
with
20% Me0H/CHC13 (2 x 50 mL), and the combined organic phase was dried over
MgSO4, filtered, and concentrated in vacuo. The residue was taken up in CHC13
(4
mL), loaded onto a silica gel column and eluted with 45% ehtyl
acetate/methylene
chloride over 1296 mL to afford imidazole M9.3f as an orange solid (261.7 mg).
1H
NMR (400 MHz, DMSO-d6) 6 ppm 12.08 (s, 0.5H), 11.81 (s, 1.5H), 7.64 (d, J =
8.3
Hz, 3H), 7.55 (d, J = 8.3 Hz, 1H), 7.41 (d, J = 1.7 Hz, 1.7H), 7.19-1.17 (m,
1.3H),
7.11 (d, J = 8.3 Hz, 3H), 4.59 (app br s, 2H), 3.41 (app br s, 2H), 2.37-2.14
(m, 6H),
1.68-1.57 (m, 2H), 1.48-1.40 (m, 2H), 1.40-0.95 (br s, 18H), 0.79-0.69 (m,
2H), 0.59-
0.48 (m, 2H). LC/MS: Anal. Calcd. for [M+H]+ C41H49 N6 0 4: 689.38; found
689.43.
Example M9.3, Step g
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H
N \ / \
H )1,.._N
45 H
A solution of carbamate M9.3f (0.2391 g, 0.347 mmol) in 25% TFA/CH2C12
(1.7 mL) was stirred at ¨25 C for lh. The volatile component was removed in
vacuo to afford the TFA salt of pyrrolidine M9.3g as a tan foam (328 mg). 1H
NMR
(400 MHz, DMSO-d6) 6 ppm 10.01 (br s, 2H), 7.70-7.68 (m, 6H), 7.20 (d, J = 8.6
Hz,
4H), 4.63-4.58 (m, 2H), 3.37-3.33 (m, 2H), 2.51-2.42 (m, 6H), 2.24-2.20 (m,
2H),
1.93-1.86 (m, 2H), 1.51-1.47 (m, 2H), 1.16-1.09 (m, 2H), 0.084-0.78 (m, 2H).
LC/MS: Anal. Calcd. for [M+H]+ C31H33 N6: 489.28; found 489.26.
Example M9.3
To a solution of pyrrolidine M9.3g/TFA salt (0.0834 g, 0.088 mmol), (S)-2-
(methoxycarbonylamino)-3-methylbutanoic acid (0.034 g, 0.194 mmol), and N,N-
diisopropylethylamine (0.123 mL, 0.706 mmol) in DMF (1.5 mL) was added HATU
(0.070 g, 0.185 mmol), and the mixture was stirred at 25 C for 4h. The
reaction
mixture was diluted with Me0H (2.5 mL) and submitted to a reverse phase HPLC
purification (XTERRA, 30X100 mm, S5; Me0H/H20/TFA) to afford Example M9.3
as an off-white foam (58 mg). 1H NMR (400 MHz, DMSO-d6) 6 ppm 14.90-14.20
(bs, 3H), 8.00 (br s, 2H), 7.68 (d, J = 8.3 Hz, 4H), 7.34 (d, J = 8.6 Hz, 4H),
7.25 (d, J
= 8.6 Hz, 2H), 5.00-4.96 (m, 2H), 4.42-4.39 (m, 2H), 3.79-3.69 (m, 2H), 3.54
(s, 6H),
2.38-2.31 (m, 5H), 2.16-2.07 (m, 2H), 1.95-1.88 (m, 2H), 1.61-1.58 (m, 2H),
0.97-
0.90 (m, 8H), 0.80 (d, J = 6.5 Hz, 8H). Rt = 1.96 min (Cond. 3); >95%
homogeneity
index (Cond. 9 and 10); LC/MS: Anal. Calcd. for [M+H]+ C45H55 N8 06: 803.42;
found 803.43.
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Example M9.4
¨0 H
¨ N-TCN - IT__.)
0.--Ns.____fH0 \ \ /
Example M9.4 was prepared from pyrrolidine M9.3g/TFA salt and (S)-2-
(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid according to
the
same procedure used for the preparation of Example M9.3. 1H NMR (400 MHz,
DMSO-d6) 6 ppm 15-14 (bs, 6H), 7.98 (br s, 2H), 7.66 (d, J = 8.3 Hz, 4H), 7.34-
7.32
(m, 5H), 4.98-4.94 (m, 2H), 4.50-4.46 (m, 2H), 3.84-3.78 (m, 6H), 3.54 (s,
6H), 3.32-
3.18 (m, 4H), 2.39-2.29 (m, 4H), 2.11-1.97 (m, 2H), 1.97-1.88 (m, 2H), 1.60-
1.57 (m,
2H), 1.51-1.26 (m, 8H), 0.99-0.87 (m, 2H), 0.84-0.72 (m, 2H). Rt = 1.77 min
(Cond.
3); >95% homogeneity index (Cond. 9 and 10); LC/MS: Anal. Calcd. for [M+H]+
C49H59N808: 887.45; found 887.50.
Examples M9.5 and M9.6
Cf----\ 0
.44,.. H 0---
Examples M9.5 and M9.6 were prepared as TFA salts starting from
dibromide M9.3b2 according to the procedures described for the preparation of
Examples M9.3 and M9.4 from the corresponding stereoisomer dibromide M9.3b1.
Example R Rt (Cond. 3); homogeneity index (Cond. 9
and 10); LC/MS data
M9.51.98 min; >95%; LC/MS: Anal. Calcd. for
\)-6)
[M+H]+ C45H55N806: 803.42; found 803.43
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M9.6 1.77 mm; >95%; LC/MS: Anal. Calcd. for
[M+H]+ C49H59N808: 887.45; found 887.60
0
Example M9.7
'0 H
N / N
HN---f0
H 0,
Example M9.7 (TFA salt) was prepared from dibromide M9.3d and acid
M12.2c according to the procedures described for the preparation of Examples
M9.3.
1H NMR (400 MHz, DMSO-d6) 6 ppm 14.90-14.42 (br s, 4H), 8.02 (br s, 2H), 7.70
(d, J = 8.5 Hz, 4H), 7.36 (d, J = 8.6 Hz, 4H), 7.25 (d, J = 8.3 Hz, 2H), 5.07
(dd, J =
10.7 Hz/7.1 Hz, 2H), 4.18-4.13 (app t, 2H), 4.12-4.08 (app t, 2H), 3.55 (s,
6H), 3.42-
3.37 (app t, 2H), 2.49-2.39 (m, 2H), 2.37-2.31 (app t, 2H), 2.02-1.93 (m, 2H),
1.87-
1.78 (app q, 2H), 1.62-1.59 (app t, 2H), 1.14 (d, J = 6.3 Hz, 6H), 0.82 (d, J
= 6.5 Hz,
6H), 0.77 (d, J = 6.8 Hz, 6H). Rt = 3.12 min (Cond. 10d); >95% homogeneity
index
(Cond. 9 and 10); LC/MS: Anal. Calcd. for [M+H]+ C45H59 N8 06: 807.46; found
807.45.
Example M9.8
0
HN--0
H
0,
Example M9.7 (TFA salt) was prepared from dibromide M9.3d and acid
M12.2c according to the procedures described for the preparation of Examples
M9.4.
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Rt = 3.01 min (Cond. 10d); >95% homogeneity index (Cond. 9 and 10); LC/MS:
Anal. Calcd. for [M+H]+ C49H63N808: 891.48; found 891.52.
Example M10
H
¨ N .ss.---N
Me02CHN J.D--(\ / )¨( --CIII\I ......5.7-1CO2Me
N-......ss N 0
Me' ,.c...._ H
Example M10, Step a
H3C.4N"\--"ISP----
I 0
Boc
The title compound, containing a diastereomeric impurity, was prepared in 5
steps from (S)-methyl 5 ¨oxopyrrolidine-2-carboxylate by employing the
procedure
described J. Med. Chem., 49:3520-3535 (2006) for the synthesis of its ethyl
ester
analog. 1H NMR (CDC13, 400 MHz): 4.35 (m, 0.5H), 4.25 (m, 0.5H), 4.05 (m,
0.5H), 3.90 (m, 0.5H), 3.73 (s, 3H), 2.20 (m, 1H), 2.00 (m, 2H), 1.65 (m, 1H),
1.50/1.40 (two overlapping br s, 9H), 1.31 (d, J = 6.0, 3H).
Example M10, Step b
OH
H3C'''4Nr"-""c(
1 0
Boc
A solution of lithium hydroxide (0.23g, 9.62 mmol) in water (5 mL) was
added dropwise to a solution of ester MlOa (1.8g, 7.4 mmol) in ethanol (10
mL), and
stirred at room temperature for 17 hr. Most of the solvent was evaporated, and
the
residue was diluted with water, 1 N HC1 was added dropwise to bring it to pH
3. It
was extracted with ethyl acetate (20 mL, 4x), dried (Na2504) and evaporated in
vacuo to afford a colorless oil, which yielded crystals when dissolved in
Et0Ac/hexanes solvent system and allowed to stand at room temperature. The
white
solid was filtrated and dried in vacuo (1.42g). 1H NMR (CDC13, 400 MHz, d =
7.24
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ppm): 4.35 (m, 1H), 3.95 (m, 1H), 2.35 (m, 1H), 2.05 (m, 2H), 1.70 (m, 1H),
1.50 (br
s, 9H), 1.25 (d, J = 7.1, 3H).
Example M10, Step c
0
H3C=1"N"\-""e 0 Br
1 0
Boc
To a solution of acid MlOb (2.16 g, 9.42 mmol) and 2-bromo-1-(4-
bromophenyl)ethanone (2.62 g, 9.42 mmol) in acetonitrile (50 mL) was added
slowly
diisopropylethylamine (1.645 mL, 9.42 mmol), and the reaction mixture was
stirred
at room temperature for 5 hr. Solvent was removed in vacuo, and the residue
was
partitioned between ethyl acetate and water (1:1, 100 mL). The organic layer
was
washed with sat. NaHCO3, dried (Na2504) and concentrated in vacuo to afford
ketoester MlOc as white solid (3.9g), which was used in the next step without
further
purification. 1H NMR (400 MHz, DMSO-d6, 6 = 2.5 ppm): 7.92 (d, J = 8.3, 2H),
7.78 (d, J = 8.5, 2H), 5.6-5.4 (m, 2H), 4.35 (m, 1H), 3.85 (m, 1H), 2.25 (m,
1H), 2.05
(m, 2H), 1.60 (m, 1H), 1.5/1.4 (two overlapping br s, 9H), 1.18 (d, J = 6.6,
3H).
LC/MS: Anal. Calcd. for [M+Na]+ C19H2481BrNaN05: 450.07; found: 450.00.
Example M10, Step d
c?:\iloc \ . Br
N
H
H3C
To a solution of ketoester MlOc (3.9 g, 9.15 mmol) in xylene (60 mL) in a
500 mL pressure tube, ammonium acetate (7.05 g, 91 mmol) was added. The
reaction vessel was sealed and heated at 140 C for 5 hr. The solvent was
removed in
vacuo, and the residue was partitioned between CH2C12 (100 mL) and water (100
mL). The organic layer was washed (Sat. NaHCO3), dried (Na2504), and
evaporated
in vacuo. The resulting crude material was purified with flash chromatograph
(30-
100% Et0Ac/Hexane) to afford bromide MlOd as a brown foam (3.0 g, yield 81%).
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1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 11.77 (s, 1H), 7.70 (d, J = 8.5, 2H),
7.52 (br s, 1H), 7.50 (d, J = 8.5, 2H), 4.80 (m, 1H), 3.85 (m, 1H), 2.10 (m,
3H), 2.70
(m, 1H), 1.5/1.3 (overlapping br s, 9H), 1.20 (m, 3H). LC/MS: Anal. Calcd. for
[M+H]+ C19H2579BrN302: 406.11; found: 406.18.
Example M10, Step e
H ----.11Me
N-......,` N
MeI'< H
Pyrrolidine M10e (HC1 salt) was prepared bromide MlOd according to the
procedure described for the synthesis of pyn-olidine Mlb (HC1 salt) from
bromide
Dlb.
Example M10
Example M10 (TFA salt) was prepared from bromide pyrrolidine M10e (HC1
salt) according to the procedure described for the synthesis of Example M1
from
pyn-oldine M3i (HC1 salt). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 8.05 (s,
2H), 7.85 (d, J = 8.1, 4H), 7.69 (d, J = 7.6, 4H), 7.56 (d, J = 8.0, 2H), 5.5
(m, 0.4H),
5.0 (m, 1.6H), 4.75 (1.6H), 4.10 (m, 0.4H), 3.95 (m, 2H), 3.50 (s, 6H), 2.50-
2.30 (m,
4H), 2.10 (m, 2H), 1.85 (m, 4H), 1.46 (d, J = 6.6, 5H), 1.20 (d, J = 6.8, 1H),
0.9-0.9
(m, 7.2H), 0.74 (d, J = 6.8, 4.8H). LC (Cond. 9 and 10): >95% homogeneity
index.
LC/MS (Cond. 3): Rt = 2.03 min. LC/MS: Anal. Calcd. for [M+H]+ C44H551\1806:
791.42; found: 791.39.
Examples M11-M12
The TFA salts Examples M11-M12 were prepared according to the procedure
described for Example M10 and by employing appropriate acids.
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H
a¨R
0
Example 0 Rt (Cond. 3); % homogeneity index (Cond. 9 and
R
10); LC/MS data
Mll o/ 1.89 min.; >98%; LC/MS: Anal. Calcd. for [M+H]+
C42H51N806: 763.39; found 763.40.
HN; 0
M12 o/ 1.82 min.; >98%; LC/MS: Anal. Calcd. for [M+H]+
/(:) C481-159N808: 875.45; found 875.42.
HN. 0
.r=Prj
0
Example M12.1
H
\)_( _______________________________________________ X_<_1_ 11µ NHCO2Me
Me02CHN N
N,.=\ N 0
0
To a solution of pyrrolidine M10e/HC1 salt (119 mg, 0.191 mmol), (S)-2-
(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid (49.0 mg, 0.226
mmol) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (40.9 mg, 0.233
mmol) in DMF (5 mL) was added DIEA (0.200 mL, 1.147 mmol) and HATU (148
mg, 0.390 mmol), and the mixture was stirred at room temperature for 1 hr.
Solvent
was removed in vacuo and the residue was dissolved in methanol and purified
with a
reverse phase HPLC (Me0H/TFA/Water) to separate the statistical mixture of
products, one of which was Example M12.1 (TFA salt; light yellow foam; 55 mg).
1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 8.09-7.82 (m, 6H), 7.68-7.55 (m, 6H),
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5.00 (m, 2H), 4.64 (app t, 2H), 4.21-3.75 (m, 6H), 3.53 (m, 6H), 3.17 (m, 2H),
2.33-
2.21 (m, 6H), 1.85 (m, 3H), 1.62-1.17 (overlay of 'X and `m', J for 'd' = 6.6
Hz,
9H), 0.88 (m, 3.6H), 0.74 (d, J = 6.8 Hz, 2.4H). LC (Cond. 9 and 10): >95%
homogeneity index. LC/MS (Cond. 10h): Rt = 1.92 min. LC/MS: Anal. Calcd. for
[M+H]+ C46H57N807: 833.44; found: 833.40.
Example M12.2
Me02CHN 0 H
N
H 1\1HCO2Me
Example M12.2, Step a
s.
0 0 s4 0 ==
z z z NO
0 Boc
0 Boc 0 Boc
M12.2a-1 M12.2a-2 M12.2a-3
The above three esters were prepared from (S)-1-tert-butyl 2-methyl 5-
oxopyrrolidine-1,2-dicarboxylate according to the procedure described in
Tetrahedon
Letters, 2003, 3203-3205.
Example M12.2, Step b
0
z
0 Boc
Borane-methyl sulfide complex (5.44 mL, 10.88 mmol) was added to a
solution of ester M12.2a-2 (1.4 g, 5.44 mmol) in THF (25 mL), and the reaction
mixture was heated at 40 C for 7 hr. The volatile component was removed in
vacuo
and the residue was partitioned between Et0Ac and water (50 mL each). The
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aqueous layer was extracted with Et0Ac (30 mL), and the combined organic phase
was dried with Na2SO4, and concentrated in vacuo. The resultant colorless oil
was
purified with a flash chromatography (0-50% Et0Ac/Hexane) to afford ester
M12.2b
as a colorless oil (0.77 g). 1H NMR (CDC13, 6 = 7.24 ppm, 400 MHz): 4.29-4.18
(m,
1H), 3.78-3.66 (m, 4H), 2.99 (app t, J = 10.1, 1H), 2.43-2.97 (m, 1H), 2.43-
2.37 (m,
1H), 2.30-2.18 (m, 1H), 1.60-1.52 (m, 1H), 1.47/1.42 (two 's', 9H), 1.08-1.05
(m,
3H).
Example M12.2, Step c
.s,
õ õs' )
Y
0 Boc
To a solution of ester M12.2b (1.69 g, 6.95 mmol) in ethanol (10 mL) was
added solution of LiOH (0.250 g, 10.42 mmol) in water (5.00 mL), and the
reaction
mixture was stirred at room temperature for 5 hr. The organic solvent was
evaporated in vacuo and the residue was diluted with water (10 mL) and washed
with
ether (10 mL). It was chilled in ice-water bath, and acidified to a pH range
of ¨2
with 1N HC1. It was then extracted with Et0Ac (20 mL, 3x). The organic layer
was
dried with Na2504 and concentrated in vacuo to afford acid M12.2c as a
colorless oil,
which became a white solid upon extended exposure to high vacuum (1.38g). 1H
NMR (CDC13, 6 = 7.24 ppm, 400 MHz): 4.39-4.22 (m, 1H), 3.80-3.69 (m, 0.91H),
3.59-3.35 (m, 0.18H), 3.03-2.89 (m, 0.91H), 2.51-2.22 (m, 2H), 1.98-1.91 (m,
0.71H), 1.68-1.60 (0.29H), 1.50/1.44 (two 's', 9H), 1.09 (app m, 3H).
Example M12.2, step d
....¨_....e 0 .
Br
--N, 0
Boc
To a solution of M12.2c (1.38 g, 6.02 mmol) and 2-bromo-1-(4-
bromophenyl)ethanone (1.673 g, 6.02 mmol) in acetonitrile (35 mL) was added
DIEA (1.051 mL, 6.02 mmol). It was stirred at room temperature for 5 hrs. The
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solvent was evaporated in vacuo and water (50 mL) and Et0Ac (70 mL) were
added,
organic layer was separated and washed by sat. NaHCO3 (30 mL), dried with
Na2SO4, evaporated in vacuo to give crude M12.2d as a red oil (2.71 g), which
was
used in the next step without further purification. 1H NMR (DMSO-d6, 6 = 2.5
ppm,
400 MHz): 7.91 (m, 2H), 7.78 (d, J = 8.5 Hz, 2H), 5.60-4.90 (m, 2H), 4.29 (app
t,
1H), 3.61 (m, 1H), 2.85 (m, 1H), 2.35-1.80 (m, 2H), 1.65 (m, 1H), 1.40-1.32
(two s,
9H), 1.02 (d, J = 6.5 Hz, 3H). LC/MS: Anal. Calcd. for [M+Na]+
C19H2481BrNNa05:
450.07; found: 450.11.
Example M12.2, step e
Br . /Hi):
Boc
To a pressure tube containing a solution of ketoester M12.2d (2.57 g, 6.03
mmol) in xylene (50 mL was added ammonium acetate (4.65 g, 60.3 mmol). The
vessel was capped and heated at 140 C for 5 hrs. The volatile component was
removed in vacuo and the residue was partitioned between DCM (50 mL) and water
(40 mL). The organic layer was dried with Na2SO4, and concentrated in vacuo.
The
resulting crude mixture was purified with flash chromato graph (30-100%
Et0Ac/Hexane) to afford imidazole M12.2.e as a brown solid (1.24g). 1H NMR
(DMSO-d6, 6 = 2.5 ppm, 400 MHz): 7.69 (d, J = 8.5 Hz, 2H), 7.58-7.48 (m, 3H),
4.70 (m, 1H), 3.65 (m, 1H), 3.02(m, 1H), 2.37 (m, 1H), 2.22 (m, 1H), 1.74-1.54
(m,
1H), 1.37-1.08 (two s, 9H), 1.03 (d, J = 6.3 Hz, 3H). LC/MS: Anal. Calcd. for
[M+H]+ C19H2579BrN302: 406.11; found: 406.18.
Example M12.2, step f
f
H s.---)
\ ,N = ----N
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Pyrrolidine M12.2f (HC1 salt) was prepared bromide M12.2e according to the
procedure described for the synthesis of pyrrolidine Mlb (HC1 salt) from
bromide
Dlb.
Example M12.2
To a mixture of pyrrolidine M12.2f /HC1 salt (200 mg, 0.321 mmol) and (S)-
2-(methoxycarbonylamino)-3-methylbutanoic acid (124 mg, 0.707 mmol) in DMF (5
mL) were added DIEA (0.337 mL, 1.928 mmol) and HATU (250 mg, 0.659 mmol),
and the mixture was stirred at room temperature for 2 hrs. The volatile
component
was removed in vacuo and the residue was purified with a reverse phase HPLC
(Phenomenex-Luna 30X100mm, S10 Axia, Me0H/TFA/Water). The resultant
sample was repurified with a different reverse phase HPLC (Water-Sunfire
30X100mm S5, ACN/TFA/Water) to afford the TFA salt of Example M12.2 as a
light yellow foam (77.1 mg). 1H NMR (Methanol-d4, 6 = 3.29 ppm, 400 MHz): 7.90
(s, 2H), 7.74 (d, J = 8.6 Hz, 4H), 7.67 (d, J = 8.5 Hz, 4H), 5.18 (m, 2H),
4.30 (app t,
2H), 4.18 (d, J= 7.3 Hz, 2H), 3.63 (s, 6H), 3.38 (m, 2H), 2.63 (m, 2H), 2.51
(m, 2H),
1.98 (m, 2H), 1.81 (m, 2H), 1.21 (d, J = 6.3 Hz, 6H), 0.83-0.90 (m, 12H). LC
(Cond.
9 and 10): >95% homogeneity index. LC/MS (Cond. 10h): Rt = 2.01 min. LC/MS:
Anal. Calcd. for [M+H]+ C44H55 N806: 791.42; found: 791.46.
Example M12.3
Me02CHNI r,
\x_sa
0 H 1\1HCO2Me
Example M12.3 (TFA salt) was prepared from pyrrolidine M12.2f /HC1 and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid according
to
the procedure described for the preparation of Example M12.2. RT: LC (Cond. 9
and
10): >95% homogeneity index. LC/MS (Cond. 10h): Rt = 1.89 min. LC/MS: Anal.
Calcd. for [M+H]+ C48H59N808: 875.45; found 875.42.
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Example M12.4-M12.5
CI H
Me02CHN. n
N ===---1\1
)13;
1LN ¨ 0
H
NHCO2Me
Example M12.4 (R = H)
Example M12.5 (R = Cl)
To a solution of Example M12.2 (free-based using MCX column and 2N
NH3/Me0H) (80 mg, 0.101 mmol) in DMF (5 mL) was added NCS (17.56 mg, 0.131
mmol), and heated at 50 C for 3 hrs. Additional NCS (5 mg, 0.037 mmol) was
added to the mixture and heating was continued for 5 more hours. The volatile
component was removed in vacuo and the residue was dissoled in Me0H and
submitted to a reverse phase HPLC condition (Me0H/TFA/Water) to afford Example
M12.4/TFA salt (light yellow foam, 24 mg) and Example M12.5/TFA salt (light
yellow foam, 28 mg). Example M12.4/TFA salt: 1H NMR (DMSO-d6, 6 = 2.5 ppm,
400 MHz): 8.17(s, 1H), 7.83-7.73 (m, 6H), 7.66 (d, J = 8.6 Hz, 2H), 7.30 (d, J
= 8.3
Hz, 1H), 7.22 (d, J = 8.3 Hz, 1H), 5.05 (m, 1H), 4.85 (m, 1H), 4.16-4.02 (m,
4H),
3.52 (s, 6H), 3.37 (m, 1H), 3.23 (m, 1H), 2.54-2.24 (m, 4H), 1.87-1.63 (m,
4H), 1.13-
1.08 (m, 6H), 0.85-0.74 (m, 12H). LC (Cond. 9a.1 and 10a.1): >95% homogeneity
index. LC/MS (Cond. 11): Rt = 4.26 min. LC/MS: Anal. Calcd. for [M+H]+
C44H54C1N808: 825.39; found: 825.50. Example M12.5/TFA salt: (DMSO-d6, 6 =
2.5 ppm, 400 MHz): 7.79 (d, J = 8.4 Hz, 4H), 7.66 (d, J = 8.4 Hz, 4H), 7.22
(d, J =
8.3 Hz, 2H), 4.85 (m, 2H), 4.13 (app t, 2H), 4.04 (app t, 2H), 3.53 (s, 6H),
3.24 (app
t, 2H), 2.39 (m, 2H), 2.26 (m, 2H), 1.90 (m, 2H), 1.66 (m, 2H), 1.09 (d, J =
6.5 Hz,
6H), 0.85 (d, J = 6.8 Hz, 6H), 0.80 (d, J = 6.5 Hz, 6H). LC (Cond. 9a and
10a.1):
>95% homogeneity index. LC/MS (Cond. 11): Rt = 4.44 min. LC/MS: Anal. Calcd.
for [M+H]+ C44H53C12N806: 859.35; found: 859.30.
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Example M12.6-M12.7
o/
H/0
0 CI \41
N 0 =
0 H
Example M12.6 (R=H) 0
Example M12.7 (R=CI)
Example 12.6 - 12.7 were prepared as TFA salts from Example M12.3
according to the procedure described for the preparation of Example 12.4 -
12.5.
Example RT (Cond. 12); % homogeneity index (Cond. 12); MS data
13.68 min.; >95%; LC/MS: Anal. Calcd. for [M+H]+ C48H58C1N808:
M12.6 909.41; found 909.8
17.12 min; >95%; LC/MS: Anal. Calcd. for [M+H]+
Example M12.8
Me02CHN 0
H NHCO2Me
Example M12.8, step a
Me02CHN 0 H
N
H
To a mixture of pyrrolidine M12.2f/HC1 salt (120 mg, 0.193 mmol) and (S)-2-
(methoxycarbonylamino)-3-methylbutanoic acid (35.1 mg, 0.200 mmol) in DMF (2
mL), were added DIEA (0.168 mL, 0.964 mmol) followed by HATU (70.4 mg, 0.185
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mmol), and the mixture was stirred at room temperature for 1 hr. The volatile
component was removed in vacuo and the residue was dissoled in Me0H and
submitted to a reverse phase HPLC condition (Me0H/TFA/Water) to isolate the
TFA
salt of pyrrolidine M12.8a as yellow solid (36mg). The sample was free based
(MCX
column; Me0H wash; 2 N NH3 /Me0H elution) to afford a yellow solid (22.3 mg).
LC/MS: Anal. Calcd. for [M+H]+ C37H44N703: 634.35; found: 634.35.
Example M12.8
To a solution of pyrrolidine M12.8a (22.3 mg, 0.035 mmol) and (S)-2-
(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid (12.99 mg,
0.060
mmol) in DMF (2 mL) were added DIEA (0.018 mL, 0.106 mmol) and HATU (13.65
mg, 0.036 mmol), and the mixture was stirred at room temperature for 2 hrs.
The
volatile component was removed in vacuo and the residue was dissoled in Me0H
and
submitted to a reverse phase HPLC condition (Column: Phenomenex-Luna
30X100mm, S10 Axia; Me0H/TFA/Water). The resulting sample was repurified
with a second reverse phase HPLC (Column: Water-Sunfire 30X100mm S5;
ACN/TFA/Water) to afford the TFA salt of Example M12.8 as a light yellow foam
(16 mg). 1H NMR (Methanol-d4, 6 = 3.29 ppm, 400 MHz): 7.92 (d, J = 5.0 Hz,
2H),
7.77 (d, J = 8.5 Hz, 4H), 7.72-7.69 (m, 4H), 5.21 (m, 2H), 4.39-4.22 (m, 4H),
3.92
(m, 2H), 3.67 (s, 6H), 3.47-3.28 (m, 4H), 2.68 (m, 2H), 2.54 (m, 2H), 2.06-
1.85 (m,
4H), 1.25 (d, J = 6.5 Hz, 6H), 0.93 (d, J = 6.7 Hz, 3H), 0.91 (d, J = 7.3 Hz,
6H). LC
(Cond. 9 and 10): >95% homogeneity index. LC/MS (Cond. 10d): Rt = 3.10 min.
LC/MS: Anal. Calcd. for [M+H]+ C46H57 N807: 833.44; found: 833.47.
Example M12.9
0/
H
N 0 N.._....)........
H 0 icai
=Sµ. C:1.
0
/
Example M12.9, Step a
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0 Boc
To a solution of mixture of M12.2a-1 and M12.2a-2 (4.75g, 18.46 mmol) was
added Superhydride (19.20 mL, 19.20 mmol) dropwise at -50 C in a
dryice/acetone
bath for 10 min. Hunig's base (13.58 mL, 78 mmol) was added, and stirred for
10
min and DMAP (0.122 g, 0.997 mmol) was added as a solid, stirred for 15 min,
and
trifluoroacetic anhydride (2.98 mL, 21.08 mmol) was added dropwise over 15
mins
Then the dryice/acetone bath was removed, and the reaction mixture was stirred
for 4
hr while allowing it to thaw to room temperature. The reaction mixture was
washed
with water (50 mL), sat. NaC1 (30 mL), and concentrated in vacuo. The
resulting
crude material was purified with flash chromatography (8-60% Et0Ac/Hexane) to
afford ester M12.9a as a yellow oil (2.85 g). 1H NMR (CDC13, 400 MHz): 6.36
(s,
0.5H), 6.25 (s, 0.5H), 4.70-4.57 (m, 1H), 3.78 (s, 3H), 2.96 (m, 1H), 2.54 (m,
1H),
1.70 (s, 3H), 1.50 (s, 4.5H), 1.44 (s, 4.5H).
Example M12.9, Step b
,O
)rs.
0
0 I
I Boc
Boc
M12.9b-1 M12.9b-2
Diethylzinc (1.1 M in toluene, 59.1 mL, 65.0 mmol) was added dropwise over
min to a cooled (-23 C) toluene (60 mL) solution of ester M12.9a (5.23 g,
21.68
mmol), and stirred for 10 min. Chloroiodomethane (9.44 mL, 130 mmol) was added
20 dropwise over 10 min, and the reaction mixture was stirred at -21 C for
16 hr. Sat.
NaHCO3 (60 mL) was added to the reaction mixture, the cooling bath was
removed,
and the mixture was stirred for 10 min. It was then filtered, and the filter
cake was
washed with toluene (50 mL). The filterate was partitioned, and the organic
layer
was dried with Na2504, and concentrated in vacuo. The resulting crude material
was
purified with flash chromatography (2-10% Et0Ac/Hexane) to afford ester ester
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M12.9b-1 (first elute; colorless oil; 2.88 g) and ester M12.9b-2 (second
elute;
colorless oil; 1.01g). Relative stereochemical assignment was made based on
NOE
studies. Ester M12.9b-1: 1H NMR (CDC13, 400 MHz): 4.65-4.52 (m, 1H), 3.72 (s,
3H), 3.28-3.17 (m, 1H), 2.44-2.32 (m, 1H), 2.16-2.10 (m, 1H), 1.51-1.42 (two
s, 9H),
1.24 (s, 3H), 1.07 (m, 1H), 0.69-0.60 (m, 1H).
Example M12.9, Step c
'1(
r N
0 I
Boc
To a solution of M12.9b-1 (2.88 g, 11.28 mmol) in Ethanol (20 mL) was
added a solution of LiOH (0.324 g, 13.54 mmol) in water (10.00 mL), and the
mixture was stirred at room temperature for 6 hr. Most of the volatile
component
was removed in vacuo, and the residue was partitioned between water (20 mL)
and
ether (20 mL). The aqueous layer was chilled in an ice-water bath, acidified
with a
1N HC1 to a pH region of 2, and extracted with Et0Ac (30 mL, 4x). The combined
organic phase was dried with Na2504, evaporated in vacuo to give acid M12.9c
as a
sticky solid (2.55g). 1H NMR (CDC13, 400 MHz): 4.64 (m, 1H), 3.25 (appt s,
1H),
2.70-2.40 (m, 1H), 2.14 (m, 1H), 1.54-1.44 (m, 9H), 1.27 (s, 3H), 1.10-0.80
(m, 1H),
0.67 (m, 1H).
Example M12.9, Step d
--, 0 ao.
Br
1::::0.......µ0
11 0
Boc
To a solution of acid M12.9c (2.05 g, 8.50 mmol) in Acetonitrile (50 mL) was
added 2-bromo-1-(4-bromophenyl)ethanone (2.361 g, 8.50 mmol) followed by DIEA
(1.484 mL, 8.50 mmol), and the reaction mixture was stirred at room
temperature for
16 hr. Most of the volatile component was removed in vacuo, and the residue
was
partitioned between Et0Ac (50 mL) and water (50 mL). The organic layer was
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washed with sat. NaHCO3 (30 mL) and sat. NaC1 (20 mL), dried with Na2SO4, and
evaporated in vacuo to afford ketoester M12.9d as white foam (3.5g). LC/MS:
Anal.
Calcd. for [M+Na]+ C201-12481BrNNa05: 462.07; found: 461.91.
Example M12.9, Step e
N \ . Br
N
H
AL-- IV
sBoc
To a mixture of ketoester M12.9e (3.5 g, 7.99 mmol) and xylene (80 mL) in a
pressure tube was added ammonium acetate (6.16 g, 80 mmol), and the reaction
mixture was capped and heated at 140 C for 4.5 hr. The volatile component was
removed in vacuo and the residue was partitioned between DCM (70 mL) and water
(70 mL). The organic layer was washed with sat. NaHCO3 (30 mL), and
concentrated in vacuo. The resulting crude material was purified with flash
chromatograph (40-100% Et0Ac/Hexane) to afford imidazole M12.9e as a brown
solid (2.8 g). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 7.68 (m, 2H), 7.57-7.49
(m, 3H), 5.08 (m, 1H), 3.20 (m, 1H), 2.45-2.09 (M, 2H), 1.69-1.52 (m, 1H),
1.42-
1.16 (m, 12H), 0.62 (m, 1H). LC/MS: Anal. Calcd. for [M+H]+ C20H2581BrN302:
420.11; found: 420.02.
Example M12.9, Step f
Boc,N
H
N
sBoc
To a pressure tube containing a solution of bromide M12.9e (0.354 g, 1.007
mmol) and 1,2-bis(trimethylstannyl)ethyne (0.354 g, 1.007 mmol) in DMF (15 mL)
was added Pd (Ph3P)4 (0.070 g, 0.060 mmol), and the reaction mixture was
degassed
for 10 min and the reaction vessel was capped and heated at 90 C for 14 hr.
Most of
the volatile component was removed in vacuo, and the residue was partitioned
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between DCM (60 mL) and water (40 mL). The organic layer was dried with
Na2SO4, and evaporated in vacuo. The resulting crude material was purified
with
flash chromatograph (40-100% Et0Ac/hexanes) to afford alkyne M12.9f as a red
solid (0.3 g). LC/MS: Anal. Calcd. for [M+H]+ C42H49N604: 701.38; found:
701.43.
Example M12.9, Step g
1-IN4/
H _____________________________
4N HC1 in dioxane (3.90 mL, 128 mmol) was added to carbamate M12.9f (0.3
g, 0.428 mmol), and the mixture was stirred at room temperature for 5 hr. The
volatile component was removed in vacuo and the residue was dried under high
vacuum overnight to afford the HC1 salt of pyrrolidine M12.9g as a yellow
solid
(0.27 g). LC/MS: Anal. Calcd. for [M+H]+ C32H33N6: 501.28; found: 501.22.
Example M12.9
To a mixture of M12.9g/HC1 salt (60 mg, 0.093 mmol) and (S)-2-
(methoxycarbonylamino)-3-methylbutanoic acid (35.8 mg, 0.204 mmol) in DMF (2
mL) were added DIEA (0.097 mL, 0.557 mmol) and HATU (72.7 mg, 0.191 mmol),
and the reaction mixture was stirred at room temperature for 2 hr. The
volatile
component was removed in vacuo and the residue was dissoled in Me0H and
submitted to a reverse phase HPLC purification (Phenomenex-Luna 30X100mm, S10
Axia, Me0H/TFA/Water). The resulting sample was repurified with a second
reverse
phase HPLC (Water-Sunfire 30X150mm OBD, ACN/TFA/Water) to afford the TFA
salt of Example M12.9 as light yellow foam (37 mg). LC/MS (Cond. 10d): Rt =
3.08
min. LC (Cond. 9 and 10): >95% homogeneity index. LC/MS: Anal. Calcd. for
[M+H]+ C46H55N806: 815.42; found: 815.46.
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Example M13
'
Me02CHN _
H 7
_
1
¨ -NHCO2Me
Example M13, Step a
Br isBoR
N>___\JD=:s,,(
\ \
N
'SEM
(SEM regiochemistry was not determined)
To a solution of bromide M3h (1.0 g, 2.47 mmol) in DMF (25 mL) was added
sodium hydride (60%; 0.109 g, 2.72 mmol), and the reaction mixture was stirred
at
room temperature for 20 min. Then (2-(chloromethoxy)ethyl)trimethylsilane
(0.482
mL, 2.72 mmol) was added dropwise, and the reaction was stirred for 21h. The
volatile component was removed in vacuo, and the residue was partitioned
between
ethyl acetate (25 mL) and water (25 mL). The aqueous layer was extracted with
ethyl acetate (2 x 25 mL), and the combined organic phase was washed with
brine
(25 mL), dried over Mg504, filtered, and concentrated in vacuo. The residue
was
taken up in CHC13 (2 mL) and loaded on a Thomson's silica gel cartridge
eluting
with 25% ethyl acetate/hexanes to afford bromide M13a of unknown regio-
chemical
make up as light yellow foam (1.171 g). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400
MHz): 7.71 (s, 1H), 7.69 (d, J = 8.6, 2H), 7.52 (d, J = 8.6, 2H), 5.62-5.31
(br s, 1H),
5.26 (d, J = 10.8, 1H), 4.84-4.66 (app br s, 1H), 3.57-3.36 (app br s, 1H),
3.50 (t, J =
8.1, 2H), 2.44-2.20 (app br s, 2H), 1.77-1.60 (app br s, 1H), 1.52-1.23 (br s,
4H),
1.23-0.96 (br s, 5H), 0.92-0.79 (m, 2H), 0.78-0.69 (m, 1H), 0.64¨ 0.55 (app br
s,
1H), 0.00 (s, 9H). LC/MS: Anal. Calcd. for [M+H]+ C25H3779BrN303Si: 534.18;
found: 533.99.
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Example M13, Step b
/ 0 Boo,
N>3::
,C
\ \
N
'SEM
(SEM regiochemistry was not determined)
To a solution of bromide M13a (0.5542 g, 1.037 mmol) and triethylamine
(0.434 mL, 3.11 mmol) in 2-propanol (2.000 mL) and water (1 mL) in a sealed
reaction vessel was added potassium vinyltrifluoroborate (0.181 g, 1.348 mmol)
followed by 1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (0.085 g, 0.104 mmol). The reaction mixture was
flushed
with nitrogen, capped and heated at 100 C for 17h. The volatile component was
removed in vacuo, and the residue was partitioned between ethyl acetate (25
mL) and
water (25 mL), and the aqueous phase was extracted with ethyl acetate (2 x 25
mL).
The combined organic phase was dried over Mg504, filtered, and concentrated in
vacuo. The residue was taken up in CHC13(4 mL) and loaded onto a Thomson's
silica gel cartridge eluting with 25% ethyl acetate/hexanes to afford alkene
M13b as a
light yellow viscous oil (345.4 mg). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz):
7.71 (d, J = 8.3, 2H), 7.67 (s, 1H), 7.45 (d, J = 8.3, 2H), 6.72 (dd, J =
17.6,11.1, 1H),
5.8 (dd, J = 0.8, 17.6, 1H), 5.60-5.29 (app br s, 1H), 5.26 (d, J = 10.8, 1H),
5.22 (d, J
= 11.8, 1H), 4.83-4.67 (app br s, 1H), 3.59 ¨ 3.35 (app br s, 1H), 3.51 (t, J
= 8.1, 2H),
2.41-2.23 (app br s, 2H), 1.76-1.59 (app br s, 1H), 1.51-1.22 (app br s, 4H),
1.22-
0.97 (app br s, 5H), 0.93-0.80 (m, 2H), 0.80-0.71 (m, 1H), 0.65-0.56 (app br
s, 1H),
0.00 (s, 9H). LC/MS: Anal. Calcd. for [M+H]+ C27H40N303Si: 482.28; found:
482.24.
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Example M13, Step c
SEM, Boo,
N
ss. \ /
N . 110 \N N
'Boo \SEM
(SEM regiochemistry was not determined)
To a solution of alkene M13b (0.748 g, 1.552 mmol) in CH2C12 (2 mL) was
added Grubbs 2nd Generation Catalyst (0.132 g, 0.155 mmol), and stirred at
room
temperature under nitrogen for 18 hr. The reaction was loaded onto a Thomson's
silica gel cartridge eluting with 30% ethyl acetate/hexanes to afford M13c as
a tan
foam (0.50 g). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 7.74 (d, J = 8.4, 4H),
7.68 (s, 2H), 7.58 (d, J = 8.3, 4H), 7.22 (s, 2H), 5.72-5.34 (bs, 3H), 5.34-
5.21 (m,
2H), 4.94-4.63 (app br s, 3H), 3.52 (t, J = 7.8, 4H), 2.45-2.20 (app br s,
4H), 1.88-
1.57 (app br s, 2H), 1.57-1.26 (br s, 8H), 1.26-0.98 (br s, 10H), 0.93-0.80
(m, 4H),
0.80-0.69 (m, 2H), 0.69 -0.51 (app bs, 2H), 0.00 (s, 18H).
Example M13, Step d
HN \
FJr_11..D1 '"
,,N = / 110 \N NH
ii- NH
4N HC1/Dioxane (13 mL) was carefully added to a flask containing M13c
(0.4799 g, 0.513 mmol) that was cooled with an ice/water bath. A mixture of
water
(4 mL) and 12N HC1 (2 mL) was added to the above mixture and the cooling bath
was removed and stirring of the reaction mixture continued for 24h. Me0H (2
mL)
was added to the reaction, and stirring was continued for 17h. All solvents
were
removed in vacuo to afford pyrrolidine M13d (.4HC1) as a yellow/tan solid (317
mg).
1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 10.78-9.87 (very br s, ¨3H), 8.01 (s,
2H), 7.87 (d, J = 8.4, 4H), 7.71 (d, J = 8.6, 4H), 7.35 (s, 2H), 4.76 (app t,
J = 8.8,
2H), 3.43 (app t, J = 5.1, 2H), 2.64 (m, 2H), 2.55-2.52 (m, 2H), 1.95 (m, 2H),
1.16-
1.05 (m, 2H), 0.86 (m, 2H). LC/MS: Anal. Calcd. for [M+H]+ C30H31N6: 475.26;
found: 475.26.
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Example M13
The TFA salt of Example M13 was prepared from pyrrolidine M13d (.4HC1)
and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid according to the
procedure described for the preparation of Example Ml. 1H NMR (DMSO-d6, 6 =
2.5 ppm, 400 MHz): 8.14-7.95 (br s, 2H), 7.80-7.74 (m, 8H), 7.40 (s, 2H), 7.25
(d, J
= 8.3, 2H), 5.0 (app t, J = 7.3, 2H), 4.41 (app t, J = 7.6, 2H), 3.81-3.67 (m,
2H), 3.54
(s, 6H), 2.57-2.46 (overlapped with DMSO-d6) (m, 2H), 2.44-2.31 (m, 2H), 2.19-
2.05
(m, 2H), 1.98-1.87 (m, 2H), 1.02-0.89 (m, 7.5H), 0.89-0.70 (m, 8.5H). LC
(Cond. 3):
Rt = 1.92 min; LC/MS: Anal. Calcd. for [M+H]+ C44H53N806: 789.41; found:
789.46.
Examples M14-M15
Example M14 (TFA salt) was prepared from pyrrolidine M13d and
appropriate acid by employing the procedure described for the synthesis of
Example
Ml. In the case of Example M15 an equimolar mixture of (S)-2-
(methoxycarbonylamino)-3-methylbutanoic acid and (S)-2-(methoxycarbonylamino)-
2-(tetrahydro-2H-pyran-4-yl)acetic acid was employed for the coupling step and
the
resultant statistical mixture of products was separated by the HPLC technique
described for Example Ml.
R2
HN \
/ N
= = \
sRi
Example R1 R2 Rt (Cond. 3); % homogeneity
index (Cond. 9 and 10);
LC/MS data
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Example R1 R2 Rt (Cond. 3); % homogeneity
index (Cond. 9 and 10);
LC/MS data
M14 o/
o/ 1.72 min.; >98%; LC/MS: Anal.
HN/0
HNO Calcd. for [M+H]+ C48H521\1808:
0
873.43; found 873.48
rj
M151.82 min.; >98%; LC/MS: Anal.
0/ 0/
HN/0
HNO Calcd. for [M+H]+ C46H55N802:
,.......(.....if,, 831.42; found 831.47
Example Ni
F
t......_.F
Me02CHN 0
\)_(
¨ -NHCO2Me
FY
F
Example Ni, Step a
0
0 NHBoc
Br
To a suspension of 2-amino-1-(4-bromophenyl)ethanone, HC1 (4.0 g, 15.97
mmol) in DCM (50.0 mL) was added sodium bicarbonate (4.02 g, 47.9 mmol). Then
Boc-anhydride (3.89 mL, 16.77 mmol) was added to the solution and the reaction
mixture was warmed tort and stirred for 18 hrs. Then DIEA (3 mL, 17.18 mmol)
was added and the reaction mixture was stirred at rt for 2 hrs, LCMS showed
that the
reaction was complete. The reaction mixture was diluted with Et0Ac and water,
the
organic phase was washed with 5% citric acid, water and sat. NaC1, dried over
anhydrous Na2504, filtered and concentrated to yield bromide N1A (5.0 g) as a
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yellow solid, which was used in the next step without further purification. 1H
NMR
(400 MHz, Me0D) ppm 7.89 (2 H, m), 7.68 (2 H, m, J=8.53 Hz), 4.52 (2 H, s),
1.38 -
1.51 (9 H, m). LC/MS (Cond. 10d): Rt = 3.56 min. LC/MS: Anal. Calcd. For
[M+Na]+ C13H16BrNaNO3: 336.03; found: 335.97.
Example N1, Step b
0 _____________________________________ ¨ 0
¨
\
BocHN /\ / NHBoc
To a solution of carbamate Nla (2.0 g, 6.37 mmol) in DMF (5 mL) was added
1,2-bis(trimethylstannyl)ethyne (1.119 g, 3.18 mmol). The reaction mixture was
degassed, tetrakis(triphenylphosphine)palladium(0) (0.184 g, 0.159 mmol) was
added, and the mixture was heated at 90 C for 4 hrs. The crude reaction
mixture
was charged to a 90 g silica gel cartridge which was eluted with a 20 min
gradient of
0-60% Et0Ac in hexane. Alkyne Nib (0.83 g) was collected as a yellow solid.
LC/MS (Cond. 10d): Rt = 4.1 min. LC/MS: Anal. Calcd. For [M+Na]+
C28H32NaN206: 515.23; found: 515.10.
Example Ni, Step c
0 _____________________________________ _ 0
/ \ ¨
\ /
H2N NH2
To a solution of alkyne Nib (1.13 g, 2.294 mmol) in 1,4-dioxane (5 mL) was
added 4 M HC1 in dioxane (4 mL, 16.00 mmol). The reaction mixture was stirred
at
rt for 2 hrs. The reaction mixture was concentrated to dryness to yield a
yellow solid.
The solid was washed with hexane and Et0Ac, then dried to yield aminoketone
Nlc,
2 HC1 (0.508 g). 1H NMR (400 MHz, Me0D) ppm 8.10 (4 H, d, J=8.53 Hz), 7.78 (4
H, d, J=8.53 Hz), 4.64 (4 H, s). LC/MS (Cond. 10d): Rt = 1.94 min. LC/MS:
Anal.
Calcd. For [M+H]+ C18H17N202: 293.12; found: 293.07.
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Example Ni, Step d
0 0 / _ \ m
FF"\--"1(
HN NH .=1
ON0
0
To a mixture of (S)-1-(tert-butoxycarbony1)-4,4-difluoropyrrolidine-2-
carboxylic acid (0.25 g, 0.995 mmol), aminoketone Nlc, 2 HC1 (0.182 g, 0.498
mmol), HATU (0.189 g, 0.498 mmol) in DMF (2 mL) was added DIEA (0.521 mL,
2.99 mmol). The reaction mixture was stirred at rt for 3 hrs. The crude
mixture was
charged to an 80 g silica gel cartridge which was eluted with a 20 min
gradient of 0-
100% Et0Ac in hexane. ketoamide Nld (0.12 g) was collected as a yellow solid.
LC/MS (Cond. 10d): Rt = 4.08 min. LC/MS: Anal. Calcd. For [M+H]+
C38H43F4N408 : 759.29; found: 759.03.
Example Ni, Step e
Boc,
____________________________________________ HN
)¨/)¨c\N
N ¨
'Bac
In a sealed tube, a mixture of ketoamide Nld (0.12 g, 0.158 mmol) and
ammonium acetate (0.122 g, 1.582 mmol) in xylene (2 mL) was heated at 140 C
for
4 hrs. The reaction mixture was partitioned between Et0Ac and water, the
organic
layer was washed with sat. NaHCO3 and sat. NaC1, dried over anhydrous Na2504,
filtered and concentrated. The residue was dissolved in a small amount of
methylene
chloride and charged to a 40 g silica gel cartridge which was eluted with a 20
min
gradient of 0-100% Et0Ac in hexane. Imidazole Nle (0.054 g) was collected as a
yellow solid. LC/MS (Cond. 10d): Rt = 3.3 min. LC/MS: Anal. Calcd. For [M+H]+
C38H41F4N604: 721.3; found: 721.10.
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Example Ni, Step f
Hp<F
HN
\N
N ¨
H
To a solution of imidazole Nle (0.054 g, 0.075 mmol) in 1,4-Dioxane was
added 4 M HC1 in dioxane (0.5 mL, 2.0 mmol) plus a few drops of Me0H. The
reaction mixture was stirred at rt for 4 hrs, and concentrated to yield
pyrrolidine Nlf,
4 HC1 (0.055 g) as a pale yellow solid. LC/MS (Cond. 10d): Rt = 2.94 min.
LC/MS:
Anal. Calcd. For [M+H]+ C28H25F4N6: 520.2; found: 521.10.
Example Ni
To a mixture of (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (0.014
g, 0.079 mmol), pyrrolidine Nlf, 4 HC1 (0.025 g, 0.038 mmol), HATU (0.030 g,
0.079 mmol) in DMF (2 mL) was added DIEA (0.039 mL, 0.225 mmol). The
reaction mixture was stirred at rt for 18 hrs, diluted with Me0H, filtered and
purified
by reverse phase HPLC to yield TFA salt of Example Ni (0.026 g) as a white
solid.
LC (Cond. 10b and 10c): >95% homogeneity index. LC/MS (Cond. 10d): Rt = 3.11
min. LC/MS: Anal. Calcd. For [M+H]+ C42H47F4N806: 835.35; found: 835.16.
Example N2
0
H 7
0 NH 0
0 H
Example N2: (S, S) 0,
F F Example N3: (S, R)
Example N4: (R, R)
Pyrrolidine Nlf was coupled with 2-(methoxycarbonylamino)-2-(tetrahydro-
2H-pyran-4-yl)acetic acid by employing the procedure described for the
synthesis of
Example Ni. The resultant three diastereomers (TFA salts) were separated by
employing the following condition: Column = Waters-Sunfire 30 X 100 mm S5;
Start
%B = 0; Final %B = 32; Gradient time = 25 min; Stop time = 25 min; Flow Rate =
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40 mL/min; Wavelength = 220 nm; Solvent A = 0.1 % TFA in 10% MeCN/90%
water; Solvent B = 0.1 % TFA in 90% MeCN/10% water.
Example Analytical conditions
N2 LC (Cond. 10b and 10c): >95% homogeneity index.
LC/MS (Cond. 10d): Rt = 2.90 min. LC/MS: Anal.
Calcd. For [M+I-1]+ C461-151F4N808: 919.37; found:
919.41.
N3 LC (Cond. 10b and 10c): >95% homogeneity index.
LC/MS (Cond. 10d): Rt = 2.96 min. LC/MS: Anal.
Calcd. For [M+I-1]+ C461-151F4N808: 919.37; found:
919.41.
N4 LC (Cond. 10b and 10c): >95% homogeneity index.
LC/MS (Cond. 10d): Rt = 3.00 min. LC/MS: Anal.
Calcd. For [M+I-1]+ C461-151F4N808: 919.37; found:
919.41.
Example N54510
Example N5-NM (TFA salt) were prepared starting from aminoketone Nlc
and appropriate starting materials, obtained from commercial sources, by
employing
the procedures described for the synthesis of Example Ni.
R N
rr\IM /)--- /X-.....ki
H
Example R Analytical conditions
N5 LC (Cond. 10b and
10c): >95%
homogeneity index. LC/MS (Cond.
0 NI(0
0 10d): Rt = 3.05 min. LC/MS: Anal.
Calcd. For [M+I-1]+ C42H51N808:
795.38; found: 795.23.
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N6 LC (Cond. 10b and 10c): >95%
homogeneity index. LC/MS (Cond.
0 Ny0 10d): Rt = 3.23 min. LC/MS: Anal.
0
Calcd. For [M+I-1]+ C44H55N806:
791.42; found: 791.54.
N7 pH LC (Cond. 10b and 10c): 93%
..
n homogeneity index. LC/MS (Cond.
10d): Rt = 3.06 min. LC/MS: Anal.
O 7,..- ,o
H"---t Calcd. For [M+1-1]+ C42H51N808:
0-..._
795.38; found: 795.4.
N7.1
1LC (Cond. 10b and 10c): 97%
homogeneity index. LC/MS (Cond.
10d): Rt = 3.056 min. LC/MS: Anal.
Of.- 0
HN ---f
Calcd. For [M+I-1]+ C44H51N806:
0--
787.39; found: 787.4.
N7.2 OH LC (Cond. 10b and 10c): 98%
;--- homogeneity index. LC/MS (Cond.
10d): Rt = 2.878 min. LC/MS: Anal.
O 7,..- ,p
H"---t Calcd. For [M+1-1]+ C42H51N808:
0-..._
795.38; found: 795.41.
N7.3 LC (Cond. 10b and 10c): > 95%
41.,7N
H homogeneity index. LC/MS (Cond.
0 Ny 10d): Rt = 3.04 min. LC/MS: Anal.
0
Calcd. For [M+I-1]+ C48F159N808:
0
875.45; found: 875.02.
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Example N7.8 & N7.9
0
'
______________________________________________ H *n
0 _ N N
NIT\ /)(
1%1N' 0 -- 0
Example N7.8: (S, S)
Example N7.9: (S, R)
When the commercially available (S)-1-(tert-butoxycarbony1)-2,5-dihydro-
1H-pyrrole-2-carboxylic acid was used in place of (S)-1-(tert-butoxycarbony1)-
4,4-
difluoropyrrolidine-2-carboxylic acid in the procedure described for the
synthesis
Example Ni, two diastereomers (Example N7.8 & N7.9) were isolated from the
final
step. The two diastereomers (TFA salts) were separated by employing the
following
condition: Column = Waters-Sunfire 30 X 100 mm S5; Start %B = 0; Final %B =
25;
Gradient time = 30 min; Stop time = 30 min; Flow Rate = 40 mL/min; Wavelength
=
220 nm; Solvent A = 0.1 % TFA in 10% MeCN/90% water; Solvent B = 0.1 % TFA
in 90% MeCN/10% water. Example N7.8: 1H NMR (400 MHz, Me0D) ppm 7.94 (2
H, s), 7.74 - 7.81 (4 H, m), 7.67 - 7.73 (4 H, m), 6.33 - 6.42 (2 H, m), 5.98 -
6.09 (4
H, m), 4.85 (2 H, d, J=1.76 Hz), 4.63 - 4.76 (2 H, m), 4.14 (2 H, d, J=8.03
Hz), 3.66
(6 H, s), 1.95 - 2.05 (2 H, m), 0.93 (6 H, d, J=6.53 Hz), 0.87 (6 H, d, J=6.78
Hz). LC
(Cond. 10d): Rt = 2.933 min; LC/MS: Anal. Calcd. for [M+H]+ C42H46N806:
758.35; found: 759.41. Example N7.9: 1H NMR (400 MHz, Me0D) ppm 7.92 (2 H,
d, J=14.81 Hz), 7.75 - 7.83 (4 H, m), 7.69 (4 H, dd, J=8.66, 2.13 Hz), 6.37 (2
H, td,
J=6.65, 2.26 Hz), 6.03 - 6.09 (1 H, m), 5.98 - 6.03 (3 H, m), 4.85 (2 H, d,
J=1.76 Hz),
4.72 (1 H, d, J=5.02 Hz), 4.59 -4.68 (1 H, m), 4.15 (2 H, dd, J=8.03, 4.77
Hz), 3.64 -
3.69(3 H, m), 3.55 - 3.62 (3 H, m), 1.95 - 2.11(2 H, m), 1.03(6 H, d, J=6.53
Hz),
0.93 (3 H, d, J=6.78 Hz), 0.87 (3 H, d, J=6.78 Hz). LC (Cond. 10d): Rt = 2.920
min;
LC/MS: Anal. Calcd. for [M+H]+ C42H46N806: 758.35; found: 759.34.
Example N8
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_______________________________________________ H nF
0
Me0
H
Example N8, step a
¨ N N
H
Pyrrolidine N8a (HC1 salt) was prepared staring from aminoketone Nlc and
(1S,3S,5S)-2-(tert-butoxycarbony1)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid
(for
its preparative method, see W02004/052850) according to the procedures
described
for the preparation of pyrrolidine Nlf. 1H NMR (Me0D, 6 = 3.33 ppm, 400 MHz):
7.98 (s, 2H), 7.87 (d, J = 8.4, 4H), 7.69 (d, J = 8.4, 4H), 5.59 (dd, J =
10.2, 5.4, 2H),
3.61-3.56 (m, 2H), 3.05-2.98 (m, 2H), 2.67 (dd, J = 14.2, 5.4, 2H), 2.20-2.14
(m, 2H),
1.28-1.22 (m, 2H), 1.19-1.13 (m, 2H). LC/MS: Anal. Calcd. for [M+H]+ C301-
129N6:
473.24; found 473.2.
Example N8
Example N8 was prepared from pyrrolidine N8a according to the procedure
described for the preparation of Example Ni with the exception that the
purification
was conducted under a modified reverse phase HPLC condition (ACN/water/
NH40Ac). Example N8 was retrieved as a pale-yellow solid. LC (Cond. 10e): >99%
homogeneity index. LC/MS (Cond. 100: Rt = 2.21 min. 1H NMR (DMSO, 6 = 2.50
ppm, 400 MHz): 12.33 (s, 0.28H), 11.86 (s, 1.72H), 7.74 (d, J = 8.4, 3.42H),
7.65 (d,
J= 8.4, 0.58H), 7.55 (d, J= 8.4, 0.58H), 7.51 (s, 2H), 7.49 (d, J= 8.4,
3..42H), 7.31
(d, J = 8.8, 2H), 5.39 (app d, 0.15H), 3.34 (dd, J = 10.8, 2.4, 3.85H), 4.29
(app t, 2H),
3.90 (br m, 2H), 3.55 (s, 6H), 2.64-2.56 (m, 2H), 2.17 (app d, 2H), 2.08-1.98
(m, 2H),
1.79-1.74 (m, 4H), 0.92-0.83 (m, 14H). LC/MS: Anal. Calcd. for [M+H]+
C44H51N806: 787.39; found 787.8.
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Example N9
0
nF
_______________________________________________ N N H OMe
N o N N
Me0 H
I H
0
Compound N9 (TFA salt) was prepared from pyrrolidine N8a and (S)-2-
(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid according to
the
procedure described for the preparation of Example Ni with the exception that
ACN/water/TFA solvent system was employed for the purification step. LC (Cond.
10e): >97% homogeneity index. LC/MS (Cond. 10g): Rt = 1.82 min. LC/MS: Anal.
Calcd. for [M+H]+ C48H55N808: 871.41; found 871.6.
Example NiO
_______________________________________________ H nMe
Me0c)[sli N5>--( 01
Me H
Example NiO, Step a
MeOya
N 0
0
A solution of (S)-5-oxopyrrolidine-2-carboxylic acid (20 g, 154 mmol) in
Me0H (140 mL) was cooled to 0 C. Then thionyl chloride (11.2 mL, 154 mmol)
was added drop wise and stirred for 30 minutes. The reaction mixture was
brought to
RT and stirred for 3 h. Then the reaction mixture was concentrated under
reduced
pressure, the resulting residue was dissolved in Et0Ac (200 mL) and the
organic
layer was washed with NaHCO3 solution. The aqueous phase was extracted with
Et0Ac (2 x 100 mL), and the combined organic layer was washed with brine,
dried
over Na2504 and concentrated in vacuo to obtain crude ester N10a (8.0 g),
which was
used as such in the next step. 1H NMR (DMSO-d6, 6 = 2.50 ppm, 400 MHz): 6 7.99
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(s, 1H), 4.21-4.17 (m, 1H), 3.67 (s, 3H), 2.39-2.29 (m, 1H), 2.17-2.09 (m,
2H), 2.02-
1.95 (m, 1H).
Example N10, Step b
MeOyN-3----
N 0
Boc
0
A solution of ester N10a (6.5 g, 45.4 mmol) and DMAP (0.277 g, 2.2 mmol)
in toluene (32 mL) was cooled to 0 C. Then (Boc)20 (10.69 mL, 46.6 mmol) was
added drop wise, and the mixture was stirred for 15 minutes at 0 C and at RT
for 12
h. The reaction mixture was quenched with NaHCO3 solution and stirred for 45
minutes. The separated organic layer was washed with water and followed by
brine,
dried over Na2504 and concentrated in vacuo. The resulting crude material was
purified by flash chromatography (Silica gel, Et0Ac: petroleum ether, 40:60)
to
afford carbamate NlOb (7 g). 1H NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 4.62
(dd,
J = 9.4, 3.0, 1H), 3.78 (s, 3H), 2.68-2.59 (m, 1H), 2.53-2.45 (m, 1H), 2.37-
2.27 (m,
1H), 2.06-1.98 (m, 1H), 1.48 (s, 9H).
Example N10, Step c
Me0y:
N OMe
Boc
0
A solution of carbamate 10b (7 g, 28.8 mmol) in THF (25 mL) was cooled to
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ppm, 400 MHz): 6 5.30-5.13 (m, 1H), 4.38-4.23 (m, 1H), 3.74/3.71/3.70 (s, 3H),
3.43/3.40/3.39/3.35 (s, 3H), 2.41-2.26 (m, 1H), 2.17-1.87 (m, 3H),
1.48/1.42/1.40 (s,
9H).
Example N10, Step d
MeOyTN2'µWle
Boc
0
A solution of CuBr:Me25 (10.15 g, 49.0 mmol) in Et20 (130 mL) was cooled
to -40 C, MeMgBr (3 M in Et20, 16.45 mL, 49.0 mmol) was added drop wise, and
the reaction mixture was stirred for 45 minutes. Then the reaction mixture was
cooled to -78 C and BF3:Et20 (6.2 mL, 49.0 mmol) was added. After being
stirred
for 30 minutes, crude ether N10c (3.2 g, 12.0 mmol) in Et20 (30 mL) was added
drop
wise, and it was stirred while allowing it to thaw from -78 C to RT over 3 h,
and
stirring was continued for 1 h at RT. The reaction mixture was quenched with
saturated NH4C1/ aq.NH3 solution (1:1, 50 mL) and stirred for 1 h at RT. The
organic layer was separated and the aqueous layer was extracted with Et20 (3 x
100
mL). The combined organic phase was washed with NaHCO3 solution, water, brine,
dried over Na2504 and concentrated in vacuo. The crude was submitted to a
chiral
purification (Column = Kromasil-(R,R) Whelk-01, 30 X 250 mm, 5 n; Isocratic (n-
hexane:Et0H, 95:5); Stop time = 30 min; Flow Rate = 20 mL/min; Wavelength =
220 nm) to afford the dominant stereoisomer, trans diastereomer N10d (850 mg).
1H
NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 4.33/4.26 (dd, J = 9.0, 1.6, 1H), 4.18-
4.04
(m, 1H), 3.71/3.70 (s, 3H), 2.30-2.05 (m, 2H), 1.94-1.88 (m, 1H), 1.57-1.50
(m, 1H),
1.46/1.39 (s, 9H), 1.19/1.15 (d, J = 6.4, 3H).
Example N10, Step e
HO(N2µ Me
Boc
0
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A solution of ester N10d (700 mg, 2.88 mmol) in Et0H (4 mL) was cooled to
0 C, and LiOH (103 mg, 4.32 mmol) in water (2 mL) was added, and the mixture
was stirred for overnight. The reaction mixture was then concentrated under
reduced
pressure, the residue was dissolved in water and extracted with Et20 (20 mL).
The
aqueous phase was acidified to pH 4 with 1 N HC1 and extracted with Et0Ac (100
mL). The organic layer was washed with brine, dried over Na2SO4 and
concentrated
in vacuo to afford acid N10e (630 mg). 1H NMR (CDC13, 6 = 7.26 ppm, 400 MHz):
6
4.39 (d, J = 6.8, 0.7H), 4.28 (d, J = 8.8, 0.3H), 4.18-4.09 (m, 0.3H), 3.98-
3.93 (m,
0.7H), 2.38-2.29 (m, 1H), 2.17-2.08 (m, 2H), 1.62-1.54 (m, 1H), 1.49/1.41 (s,
9H),
1.22-1.17 (m, 3H). LC/MS: Anal. Calcd. for [M-HI C11H18N04: 228.13; found
228.2.
Example N10, Step f
H_
R N\ / (¨)___s_i--r- N,
\ , \ N R
me U H
N10f.1 (R = Boc)
N10f.2 (R = H; 4HCI salt)
Bis-carbamate N10f.1 was prepared from acid N10.e according to the
procedure described for the synthesis of bis-carbamate Nle. To a solution of
bis-
carbamate N10f.1 (320 mg, 0.47 mmol) in Me0H (6 mL) was added HC1/dioxane
(4N, 20 mL) at 0 C and the mixture was stirred for 2 h while allowing it to
thaw to
room temperature. The volatile component was removed in vacuo, and the residue
was co-evaporated with anhydrous DCM (3 x 5 mL) and dried under high vacuum to
afford pyrrolidine N10f.2 (4HC1, 300 mg) as a pale yellow solid, which was
submitted to the next step as such. 1H NMR (DMSO-d6, 6 = 2.50 ppm, 400 MHz): 6
9.76 (br s, 2H), 9.49 (br s, 2H), 7.90 (d, J = 8.4, 4H), 7.61 (d, J = 8.4,
4H), 4.97-4.91
(m, 2H), 3.94-3.81(m, 2H), 2.56-2.42 (obscured, 2H), 2.40-2.30 (m, 4H), 1.73-
1.68
(m, 2H), 1.39 (d, J = 6.4, 6H). LC/MS: Anal. Calcd. for [M+H]+ C3oH33N6:
477.27;
found 477.2.
Example N10
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HATU (106 mg, 0.280 mmol) was added to the DMF (5 mL) solution of
pyrrolidine N10E2 (84 mg, 0.136 mmol), (S)-2-(methoxycarbonylamino)-3-
methylbutanoic acid (50 mg, 0.286 mmol) and DIEA (0.1 mL, 0.546 mmol) and
stirred at room temperature for 2 h. The volatile component was removed in
vacuo
and the residue was dissolved in Et0Ac (50 mL). The organic layer was washed
with saturated NaHCO3 solution, saturated NH4C1 and the aqueous phase was
extracted with Et0Ac (25 mL). The combined extracts were washed with water,
brine, dried over Na2SO4 and concentrated in vacuo. The crude was submitted to
reverse phase HPLC purification (ACN/water/NH40Ac) to afford Example NiO as a
free base (25 mg). LC (Condition 10e and 10e.1): >97% homogeneity index.
LC/MS (Condition 10e.5):Rt= 1.63 min. 1H NMR (DMSO-d6, 6 = 2.50 ppm, 400
MHz): 6 12.21/12.08/11.83 (s, 2H), 7.78-7.74 (m, 3H), 7.67-7.47 (m, 6H), 6.98-
6.94
(m, 1H), 5.18 (d, J = 8.0, 0.2H), 5.04 (d, J = 8.8, 1.8H), 4.37-4.34 (m, 2H),
4.25-4.21
(m, 1.4H), 4.0-3.95 (m, 0.6H), 3.55 (s, 4H), 3.35-3.32 (obscured, 2H), 2.10-
1.49 (m,
10H), 1.35 (d, J = 6.4, 4H), 1.17 (d, J = 6.4, 2H), 0.95 (d, J = 6.8, 4H),
0.91 (d, J =
6.4, 2H), 0.82/0.77 (d, J = 6.8, 6H). LC/MS: Anal. Calcd. for [M+H]+
C44H55N806:
791.42; found 791.4.
Example N11
0 (03
1._..._n0 ____________________________________ H
\)__(¨/x_NI
Me0 H N ¨.711"
0
Example N11 (TFA salt) was prepared from pyrrolidine if (4HC1 salt) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid according
to
the procedure described for Example N10 with the exception that ACN/water/TFA
was used for the HPLC purification step. LC (Condition 10e and 10e.1): >98%
homogeneity index. LC/MS (Condition 10e.5):Rt= 1.44 min. LC/MS: Anal. Calcd.
for [M+H]+ C48H59N808: 875.44; found 875.4.
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Example N11.1 &N11.2
Me0 ¨
¨ N N H OMe
H N
.s..,......
* HN
Example N11.1 (S, S)
Example N11.2 (S, R) N io->L11:1
Example 11.1, Step a
"
H
141 N ¨ N
(4HCI)
Pyrrolidine 11.1a (4HC1 salt) was prepared from (S)-5-(tert-butoxycarbony1)-
5-azaspiro[2.4]heptane-6-carboxylic acid (see Patent application #
US2009/0068140
for its preparation) according to the procedure described for pyrrolidine
Nlf.2. 1H
NMR (DMSO-d6, 6 = 2.50 ppm, 400 MHz): 6 10.26 (br s, 2H), 9.50 (br s, 2H),
7.94
(s, 2H), 7.91 (d, J = 8.4, 4H), 7.62 (d, J = 8.4, 4H), 5.09-4.99 (m, 2H), 3.45-
3.38 (m,
2H), 3.19-3.16 (m, 2H), 2.57-2.48 (obscured, 2H), 2.25 (d, J = 7.6, 1.1H),
2.22 (d, J =
7.2, 0.9H), 0.86-0.69 (m, 8H). LC/MS: Anal. Calcd. for [M+H]+C32H33N6: 501.27;
found 501.2.
Example N11.1 &N11.2
Example N11.1 (TFA salt) was prepared from pyrrolidine N11.1a (4HC1 salt)
and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid according to the
procedure described for Example N10. LC (Condition 10e and 10e.1): >95%
homogeneity index. LC/MS (Condition 10e.5):Rt= 1.61 min. 1H NMR (DMSO-d6, 6
= 2.50 ppm, 400 MHz): 6 8.06 (br s, 2H), 7.84 (d, J = 8.4, 4H), 7.62 (app d,
4H), 7.4
(d, J = 7.6, 2H), 5.25 (t, J = 7.6, 2H), 4.03 (app t, 2H), 3.86 (d, J = 9.4,
2H), 3.74 (d, J
= 9.4, 2H), 3.55 (s, 6H), 2.23 (d, J = 7.6, 4H), 2.02-1.98 (m, 2H), 0.94-0.89
(m, 2H),
0.84 (d, J = 6.8, 6H), 0.81 (d, J = 6.4, 6H), 0.68-0.62 (m, 6H). LC/MS: Anal.
Calcd.
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for [M+H]+ C46H55N806: 815.42; found 815.4. The diastereomer Example N11.2
(TFA salt; eluted second from column; 1H NMR analysis suggested non-symmetric
molecule) was also isolated. LC (Condition 10e): >92% homogeneity index. LC/MS
(Condition 10e.5):Rt= 1.65 min. LC/MS: Anal. Calcd. for [M+H]+ C46H55N806:
815.42; found 815.4.
Example N12
0
H
N N NH .10Me
Me0 H \ \ N 0 0
H
0
Example N12 (TFA salt) was prepared from Pyrrolidine 11.1a (4HC1 salt) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid according
to
the procedure described for Example N10. LC (Condition 10e and 10e.1): >99%
homogeneity index. LC/MS (Condition 10e.5):Rt= 1.51 min. LC/MS: Anal. Calcd.
for [M+H]+ C50H59N808: 899.44; found 899.4.
Example N13
Me
0 N \ /NI N OMe
N \)¨ __________ H
MeO) ( OCO
H
Me
Example N13, Step a
Me
MeOrN
N 0
Boc
0
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A solution of ester NlOb (5 g, 20.57 mmol) in THF (50 mL) was cooled to -
78 C, and LiHMDS (1M in THF, 20.57 mL, 20.57 mmol) was added drop wise and
the mixture was stirred for 45 minutes at same temperature. Then Mel (3.18 mL,
51.44 mmol) was added drop wise and stirred for 2 h at same temperature, and
stirring was continued under inert atmosphere while allowing the bath
temperature to
thaw to RT over 12 h The reaction mixture was quenched with AcOH (2.35 mL,
41.15 mmol)) and water (8 mL). The volatile component was removed in vacuo and
the residue was dissolved in Et0Ac (250 mL). The organic layer was separated
and
washed with water, and the aqueous phase was back extracted with Et0Ac (100
mL).
The combined organic extract was washed with brine, dried over Na2SO4 and
concentrated in vacuo. The resulting crude material was purified by flash
chromatography (Silica gel; Et0Ac: petroleum ether, 30:70) to afford the
methylated
product N13a (1.45 g,). 1H NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 4.58 (d, J =
9.6,
1H), 3.78 (s, 3H), 2.73-2.66 (m, 1H), 2.31-2.26 (m, 1H), 1.98-1.89 (m, 1H),
1.51 (s,
9H), 1.22 (d, J = 6.8, 3H).
Example N13, Step b
Me
MeOy'N':4-1
Boc
0
To a solution of ester N13a (1 g, 3.89 mmol) in THF (18 mL) was added
BH3:Me25 (3.89 mL, 7.78 mmol) drop wise under nitrogen atmosphere, and the
reaction mixture was heated at 40 C for 7 h. The volatile component was
removed
in vacuo and the residue was dissolved in Et0Ac (100 mL) and was washed with
water and the aqueous phase was back extracted with Et0Ac (50 mL). The
combined extract was washed with brine, dried over Na2504 and concentrated in
vacuo. The crude was purified by flash chromatography (Silica gel, Et0Ac:
petroleum ether, 25:75) to afford proline N13b (410 mg). 1H NMR (CDC13, 6 =
7.26
ppm, 400 MHz): 6 4.36 (dd, J = 8.8, 2.0, 0.4H), 4.26 (dd, J = 8.8, 3.2, 0.6H),
3.75-
3.64 (m, 1H), 3.71 (s, 3H), 2.99-2.87 (m, 1H), 2.42-2.36 (m, 1H), 2.09-2.03
(m, 1H),
1.88-1.80 (m, 1H), 1.46/1.40 (s, 9H), 1.033/1.027 (d, J = 6.8, 3H).
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Example N13, Step c
Me
HON)
I Boc
0
A solution of proline N13b (400 mg, 1.64 mmol) in Et0H (6 mL) was cooled
to 0 C, and LiOH (63 mg, 2.63 mmol) in water (3 mL) was added and the mixture
was stirred at RT for 5 h. It was then concentrated under reduced pressure,
and the
residue was dissolved in water and extracted with Et20 (20 mL). The aqueous
phase
was acidified to pH 4 with 1 N HC1 and extracted with Et0Ac (100 mL). The
organic phase was washed with brine, dried over Na2504 and concentrated in
vacuo
to afford acid N13c (630 mg). 1H NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 4.31
(d, J
= 7.2, 1H), 3.47 (t, J = 8.4, 1H), 2.81 (app t, 1H), 2.53-2.44 (m, 1H), 2.38-
1.80 (m,
2H), 1.47 (s, 9H), 1.02 (d, J = 6.4, 3H). LC/MS: Anal. Calcd. for [M-H]-
Ciitli8N04: 228.13; found 227.7.
Example N13, Step d
Me
_______________________________________________ H
yH (4HCI)
Me
Pyrrolidine N13d (4HC1 salt) was prepared from acid N13c according to the
procedure described for the preparartion of pyrrolidine N10f.2. 1H NMR (Me0D,
6 =
3.34 ppm, 400 MHz): 6 7.87 (s, 2H), 7.86 (d, J = 8.0, 4H), 7.65 (d, J = 8.0,
4H), 5.21-
5.18 (m, 2H), 3.62-3.58 (m, 2H), 3.12-3.07 (m, 2H), 2.83-2.78 (m, 2H), 2.69-
2.62 (m,
2H), 2.31-2.22 (m, 2H), 1.27 (d, J = 6.4, 6H). LC/MS: Anal. Calcd. for [M+H]+
C30I-133N6: 477.27; found 477.2.
Example N13
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Example N13 (TFA salt) was prepared from pyrrolidine N13d (4HC1 salt) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid according to the procedure
described for the preparation of Example N10. LC (Condition 10e and 10e.1):
>98%
homogeneity index. LC/MS (Condition 10e.5):Rt= 1.57 min. 1H NMR (DMSO-d6, 6
= 2.50 ppm, 400 MHz): 6 8.01-7.81 (m, 6H), 7.69-7.62 (m, 4H), 7.33 (d, J =
8.4, 2H),
5.22-5.16 (m, 2H), 4.09 (app t, 2H), 3.98 (t, J = 8.4, 2H), 3.56 (s, 6H), 3.39-
3.35 (m,
2H), 2.69-2.65 (m, 2H), 2.20-2.11 (m, 2H), 2.05-1.92 (m, 4H), 1.07 (d, J= 6.4,
6H),
0.87 (d, J = 6.8, 6H), 0.83 (d, J = 6.4, 6H). LC/MS: Anal. Calcd. for [M+H]+
C44H55N806: 791.42; found 791.4.
Example N14
Me me
; >
)\--N\)¨ ______________________________________ H
Me0 --(
_.... H
Me Me
Example N14, Step a
Me
Me0
N 0
Boc
0
A solution of ester NlOb (1 g, 4.11 mmol) in THF (20 mL) was cooled to -78
C, and LiHMDS (1M in THF, 8.2 mL, 8.22 mmol) was added to it drop wise and
stirred for 15 minutes. Then Mel (0.51 mL, 8.22 mmol) was added drop wise and
stirred at -78 C for 15 minutes and at RT for 2 h. The volatile component was
removed in vacuo and the residue was dissolved in Et0Ac (100 mL), and washed
with NH4C1 solution. The aqueous phase was back extracted with Et0Ac (50 mL),
and the combined extract was washed with water, brine, dried over Na2504 and
concentrated in vacuo. The crude was purified by flash chromatography (Silica
gel;
Et0Ac: petroleum ether, 30:70) to afford dimethylated product N14a (700 mg).
1H
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NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 4.53 (dd, J = 9.2, 5.2, 1H), 3.79 (s,
3H),
2.21 (dd, J = 13.2, 9.6, 1H), 1.97-1.88 (m, 1H), 1.50 (s, 9H), 1.21 (s, 6H).
Example N14, Step b
Me
_ ---Me
Me0 OH
N
Boc
0
A solution of ester N14a (1.4 g, 5.16 mmol) in THF (25 mL) was cooled to -
78 C and Superhydride (1M in THF, 6.19 mL, 6.19 mmol) was added to it drop
wise
under the N2. The reaction mixture was stirred for 30 minutes, quenched with
saturated NaHCO3 solution and allowed to warm up to 0 C. Then 30% aqueous
H202 (3 mL) was added and stirred for 20 minutes at 0 C. The reaction mixture
was
concentrated under reduced pressure and the residue was dissolved in DCM (100
mL) and washed with water. The aqueous phase was extracted with DCM (2 x 50
mL), and the combined organic extract was washed with brine, dried over Na2504
and concentrated in vacuo to afford crude N14b (1.25 g) which was used as such
in
the next step.
Example 14, Step c
Me
Me
MeOzsi----
N
Boc
0
To a solution of crude N14b (1.25 g, 5.16 mmol) in DCM (30 mL) was added
EtSiH (0.84 mL, 5.32 mmol), the reaction mixture was cooled to -78 C and
BF3:Et20 (0.73 mL, 5.83 mmol) was added drop wise under the N2. After 30
minutes
of stirring, additional EtSiH (0.84 mL, 5.32 mmol) followed by BF3:Et20 (0.73
mL,
5.83 mmol) were added. After stirring for 2 h at -78 C, the reaction mixture
was
quenched with NaHCO3 solution and extracted with DCM (2 x 100 mL). The
combined organic extract was dried over Na2504 and concentrated in vacuo. The
resultant crude material was purified by flash chromatography (Silica gel;
Et0Ac:
petroleum ether, 25:75) to afford proline N14c (470 mg). 1H NMR (CDC13, 6 =
7.26
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ppm, 400 MHz): 6 4.34/4.27 (t, J = 8.4, 1H), 3.74/3.73 (s, 3H), 3.38-3.18 (m,
2H),
2.02 (dd, J = 12.4, 8.0, 1H), 1.78-1.70 (m, 1H), 1.47/1.42 (s, 9H), 1.13/1.06
(s, 6H).
Example N14, Step d
Me
-Me
HOy.N )
N
Boc
0
A solution of proline N14c (500 mg, 1.94 mmol) in Et0H (8 mL) was cooled
to 0 C, LiOH (93 mg, 3.89 mmol) in water (4 mL) was added, and the reaction
mixture was stirred at RT for 6 h. It was then concentrated under reduced
pressure,
and the resulting crude material was dissolved in water and extracted with
Et20 (20
mL). The aqueous phase was acidified to pH 4 with 1 N HC1 and extracted with
Et0Ac (100 mL). The organic phase was washed with brine, dried over Na2504 and
concentrated in vacuo to afford acid N14d (454 mg). 1H NMR (CDC13, 6 = 7.26
ppm,
400 MHz): 6 4.42-4.23 (m, 1H), 3.26 (d, J = 10.8, 1H), 3.10 (d, J = 10.8, 1H),
2.30-
2.20 (m, 1H), 1.96-1.82 (m, 1H), 1.50/1.45 (s, 9H), 1.11/1.07 (s, 6H).
Example N14, Step e
Me me
_... H
(4HCI)
MeA¨me
Pyrrolidine N14e (4HC1 salt) was prepared from acid N14d according to the
procedure described for the preparation of pyrrolidine N10f.2. 1H NMR (DMSO-
d6, 6
= 2.50 ppm, 400 MHz): 6 10.28 (br s, 2H), 9.50 (br s, 2H), 7.91-7.55 (m, 6H),
7.30-
7.05 (m, 4H), 5.05-4.96 (m, 2H), 3.19-3.01 (m, 4H), 2.33-2.26 (m, 4H), 1.24
(s, 6H),
1.22 (s, 6H). LC/MS: Anal. Calcd. for [M+H]+ C32H37N6: 505.3; found 505.2.
Example N14
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Example N14 (free base) was prepared from pyrrolidine N14e (4HC1 salt) and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid according to the procedure
described for Example N10 with the exception that the crude was purified with
an
HPLC using CH3CN/Water/TFA solvent system and the resultant TFA salt product
was partitioned between Et0Ac and 10% NaHCO3, and the organic phase was dried
(Na2SO4) and evaporated in vacuo. LC (Condition 10e and 10e.1): >97%
homogeneity index. LC/MS (Condition 10e.5):Rt= 1.64 min. 1H NMR (DMSO-d6, 6
= 2.50 ppm, 400 MHz): 6 12.21/11.87 (s, 2H), 7.78-7.73/7.67-7.63 (m, 4H), 7.57-
7.53 (m, 2H), 7.51-7.44 (m, 4H), 7.21 (d, J = 8.0, 2H), 4.99 (t, J = 8.6, 2H),
4.01 (t, J
= 8.0, 2H), 3.78 (d, J = 9.8, 2H), 3.54/3.52 (s, 6H), 3.45 (d, J = 9.8, 2H),
2.07-1.88
(m, 6H), 1.17 (s, 6H), 1.03 (s, 6H), 0.85 (d, J = 6.8, 6H), 0.82 (d, J = 6.8,
6H).
LC/MS: Anal. Calcd. for [M+H]+ C46H59N806: 819.45; found 819.4.
Example N15
0 Me me
)
OMe
Me0 H N1N--(¨ x ¨ \ _______________________ "--%'N
0 0
Melvie
0
Example N15 (TFA salt) was prepared from pyrrolidine N14e (4HC1 salt) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid according
to
the procedure described for Example N10 with the exception that the reverse
phase
HPLC system used ACN/Water/TFA solvent. LC (Condition 10e and 10e.1): >96%
homogeneity index. LC/MS (Condition 10e.5):Rt= 1.55 min. LC/MS: Anal. Calcd.
for [M+H]+ C50H63N808: 903.47; found 903.4.
Example N16.1, N16.2 &N16.3
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Me
H* ---:¨..--
MH e
Me0)L[sil NNH __________________ \)( 0)¨SNIn¨>Lsi
0 0
Me
Example N16.1, Step a
OTf
Me0:-----S
N
Boc
0
A solution of NaHMDS (1M in THF, 13.56 mL, 13.56 mmol) in THF (25
mL) was cooled to -78 C, and a THF (15 mL) solution of (S)-1-tert-butyl 2-
methyl
4-oxopyrrolidine-1,2-dicarboxylate (3 g, 12.33 mmol) was added drop wise.
After 15
minutes of stirring, N-phenyl-bis(trifluoromethanesulfonimide) (4.4 g, 12.33
mmol)
in THF (15 mL) was added, and the reaction mixture was stirred at -78 C for 3
h. It
was then quenched with NaHCO3 solution at -78 C, and the mixture was then
stirred
at RT. The organic layer was separated and the aqueous phase was extracted
with
Et0Ac (2 x 50 mL). The combined organic extract was washed with water and
brine,
dried over Na2504 and concentrated in vacuo. The crude was purified by flash
chromatography (Silica gel; Et0Ac: petroleum ether, 15:85) to afford triflate
N16. la
(4.3 g). 1H NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 5.75-5.74/5.71-5.69 (m, 1H),
5.08-5.06/5.02-4.99 (m, 1H), 4.40-4.25 (m, 2H), 3.77/3.76 (s, 3H), 1.48/1.43
(s, 9H).
Example N16.1, Step b
Me
Me0"--=-5
N
Boc
0
A solution of CuI (1.46 g, 7.6 mmol) in THF (30 mL) was cooled to -30 C,
MeMgBr (3 M in Et20, 16.0 mmol) was added drop wise, and the reaction mixture
was stirred for 30 minutes. It was then cooled to -78 C, and triflate N16.1a
(2.4 g,
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6.4 mmol) in THF (10 mL) was added drop wise. After being stirred for 2 h, the
reaction mixture was quenched with NH4C1 solution. The organic layer was
separated and the aqueous phase was extracted with Et0Ac (2 x 100 mL). The
combined organic phase was washed with water and brine, dried over Na2SO4 and
concentrated in vacuo. The resulting crude material was purified by flash
chromatography (Silica gel; Et0Ac: petroleum ether, 15:85) to afford N16. lb
(560
mg). 1H NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 5.37/5.33 (br s, 1H), 4.99-
4.97/4.91-4.89 (m, 1H), 4.21-4.02 (m, 2H), 3.74/3.73 (s, 3H), 1.80/1.78 (s,
3H),
1.49/1.43 (s, 9H).
Example N16.1, Step c
Me
HOy(-1
N
Boc
0
A solution N16. lb (560 mg, 2.32mmol) in Et0H (10 mL) was cooled to 0 C,
and LiOH (89 mg, 3.71 mmol) in water (5 mL) was added, and the mixture was
stirred for overnight. It was then concentrated under reduced pressure, and
the resiue
was dissolved in water and extracted with Et20 (20 mL). The aqueous phase was
acidified to pH 4 and extracted with Et0Ac (100 mL). The organic layer was
washed
with brine, dried over Na2504 and concentrated in vacuo to afford acid N16.1c
of
unknown stereochemical integrity (490 mg). 1H NMR (CDC13, 6 = 7.26 ppm, 400
MHz): 6 5.57/5.37 (br s, 1H), 5.07/4.94 (br s, 1H), 4.16-4.01 (m, 2H), 1.81(s,
3H),
1.52/1.46 (s, 9H).
Example N16.1, Step d
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Me
H
Isli1.--k>N
_(/¨..\)_c()_c....ir----
N
H (4HCI)
Me
Pyrrolidine N16. id (4HC1 salt) was prepared from acid N16. 1c according to
the procedure described for the preparation of pyrrolidine N10f.2. 1H NMR
(Me0D, 6
= 3.34 ppm, 400 MHz): 6 8.09 (s, 2H), 7.88 (d, J = 8.6, 4H), 7.33 (d, J = 8.6,
4H),
6.01 (s, 2H), 5.79 (s, 2H), 4.29 (d, J = 15.2, 2H), 4.21 (d, J = 15.2, 2H),
2.07 (s, 6H).
LC/MS: Anal. Calcd. for [M+H]+ C301-129N6: 473.24; found 473.2.
Example N16.1, N16.2 &N16.3
When pyrrolidine N16.1d (4HC1) was reacted with (S)-2-
(methoxycarbonylamino)-3-methylbutanoic acid and purified according to the
procedure described for Example N10, with the exception of using
CH3CN/Water/TFA for the HPLC purification, the following three diastereomers
were isolated. Example N16.1 (TFA salt; eluted first). LC (Condition 10e and
10e.1): >97% homogeneity index. LC/MS (Condition 10e.5):Rt= 1.54 min. LC/MS:
Anal. Calcd. for [M+H]+ C44H5iN806: 787.39; found 787.4. Example N16.2 (TFA
salt; eluted second): LC (Condition 10e and 10e.1): >97% homogeneity index.
LC/MS (Condition 10e.5):Rt= 1.56 min. 1H NMR (Me0D, 6 = 3.34 ppm, 400 MHz):
6 7.92 (s, 2H), 7.89-7.77 (m, 4H), 7.72-7.66 (m, 4H), 6.03-5.97 (m, 2H), 5.68-
5.64
(m, 2H), 4.81-4.76 (m, 2H), 4.64-4.47 (m, 2H), 4.15 (d, J = 8.4, 2H), 3.68 (s,
6H),
2.01 (s, 6H), 2.00-1.92 (m, 2H), 0.94 (d, J = 6.8, 6H), 0.88 (d, J = 6.8, 6H).
LC/MS:
Anal. Calcd. for [M+H]+ C44H5iN806: 787.39; found 787.4. Example N16.3 (TFA
salt; eluted last): LC (Condition 10e and 10e.1): >92% homogeneity index.
LC/MS
(Condition 10e.6):Rt= 1.78 min. LC/MS: Anal. Calcd. for [M+H]+ C44H51N806:
787.39; found 787.4.
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Example N17.1 &N17.2
0 (0:3 Me
H ---
Me0 )__(¨X_c___,
H
....,..----
Me 0
The following two diastereomers were isolated when pyrrolidine N16.1d
(4HC1) was reacted with (S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-
yl)acetic acid and purified according to the procedure described for Example
N10
with the exception that CH3CN/Water/TFA was used for the reverse phase HPLC
purification. Example 17.1 (TFA salt; eluted first): LC (Condition 10e and
10e.1):
>94% homogeneity index. LC/MS (Condition 10e.5):Rt= 1.46 min. LC/MS: Anal.
Calcd. for [M+H]+ C48H55N808: 871.41; found 871.2. Example 17.2 (TFA salt;
eluted second): LC (Condition 10e and 10e.1): >93% homogeneity index. LC/MS
(Condition 10e.5):R= 1.48 min. LC/MS: Anal. Calcd. for [M+H]+ C48H551\1808:
871.41; found 871.4.
Example N22
Me
-7-.7
¨\ N M.10Me
)\---N HN----
Me0 H N-...}-"N _________________ ¨ ¨ \ , µ,- NH 0
0
Me
Example N22, Step a
Me
0 .Br
Br
A solution of 1-bromo-4-iodo-2-methylbenzene (5.0 g, 16.83 mmol) in
dioxane (75 mL) was purged with N2 for 10 minutes. Then PdC12(Ph3P)2 (428 mg,
0.609 mmol) and tri-n-buty1(1-ethoxyvinyl)stanane (5.94 mL, 17.59 mmol) was
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added, and the reaction mixture was purged with N2 for 10 minutes and heated
at 80
oC for overnight. Water (25 mL) was added to the reaction mixture and cooled
to 0
oC, NBS (3 g, 17.1 mmol) was added and the mixture was stirred for 30 minutes.
The volatile component was removed under reduced pressure, and water was added
to the resulting residue and it was extracted with DCM (3 x 100 mL). The
combined
organic extract was dried over Na2SO4 and concentrated in vacuo. The crude was
purified by flash chromatography (ISCO; Et0Ac: petroleum ether, 2:98) to
afford
ketone N22a (2.6 g). 1H NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 7.84 (d, J =
0.8,
1H), 7.65-7.62 (m, 2H), 4.32 (s, 2H), 2.48 (s, 3H).
Example N22, Step b
Me
0 =lgoc J.L Br
To a stirred solution of (1R,3S,5R)-2-(tert-butoxycarbony1)-2-
azabicyclo[3.1.0]hexane-3-carboxylic acid (2.23 g, 9.89 mmol) and TEA (1.3 mL,
9.80 mmol) in ACN (30 mL) was added dibromide N22a (2.6 g, 8.99 mmol) in ACN
(45 mL), and the reaction mixture was stirred for 90 minutes at room
temperature.
The volatile component was removed in vacuo, the residue was partitioned
between
water and DCM, and the organic layer was washed with brine, dried over Na2SO4
and concentrated in vacuo. The resulting crude material was purified by flash
chromatography (ISCO; Et0Ac: petroleum ether, 40:80) to afford ketoester N22b
(1.6 g). 1H NMR (DMSO-d6, 6 = 2.50 ppm, 400 MHz): 6 7.98 (s, 1H), 7.79 (d, J =
8.2, 1H), 7.71 (d, J = 8.2, 1H), 5.59-5.42 (m, 2H), 4.18-4.12 (m, 1H), 3.40-
3.32 (m,
1H), 2.43 (s, 3H), 2.41-2.37 (m, 2H), 1.68-1.60 (m, 1H), 1.42/1.32 (s, 9H),
0.78-0.73
(m, 1H), 0.56-0.51 (m, 1H). LC/MS: Anal. Calcd. for [M+H-Boc]+ C15H17BrNO3:
338.03; found 338Ø
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Example N22, Step c
Me
HN \ =
Br
14?..c....),......z.N
At4"---
To a solution of N22b (1.6 g, 3.66 mmol) in xylenes (30 mL) was added
NH40Ac (2.82 g, 36.6 mmol) and the reaction mixture was purged with N2 for 10
minutes. The reaction mixture was heated in a pressure tube at 130 C for over
night.
The volatile component was evaporated under reduced pressure and the crude was
treated with saturated NaHCO3 solution, stirred and then extracted with DCM
(200
mL). The organic layer was washed with water and brine, and dried over Na2SO4
and concentrated in vacuo. The resulting crude material was purified by flash
chromatography (ISCO; MeOH:DCM: 0.3:99.7) to afford imidazole N22c (1.1 g). 1H
NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 10.8-10.2 (br s, 1H), 7.68 (br s, 1H),
7.52-
7.40 (m, 2H), 7.23 (br s, 1H), 4.86-4.78 (m, 1H), 3.60-3.30 (m, 1H), 3.25-3.20
(m,
1H), 2.42 (s, 3H), 2.40-2.32 (m, 1H), 1.80-1.70 (m, 1H), 1.48 (s, 9H), 0.90-
0.81 (m,
1H), 0.49-0.42 (m, 1H). LC/MS: Anal. Calcd. for [M+H]+ C201-125BrN302: 418.11;
found 420.2.
Example N22, Step d
(¨/
Me '71
HN")¨ irsl."--:-"-r-610c
N \ ¨ \ / ) µ.¨NH
,s4--- Me
A solution of imidazole N22c (940 mg, 2.25 mmol) in DMF (47 mL) was
purged with N2 for 10 minutes, Pd(Ph3P)4 (98 mg, 0.084 mmol) and bis-
trimethylstanylacetylene (395 mg, 1.12 mmol) were added, and the reaction
mixture
was purged with N2 for additional 10 minutes and heated at 90 C for over
night. The
volatile component was removed under reduced pressure and the resulting crude
material was purified by flash chromatography (ISCO; Me0H :DCM, 0.5:99.5) to
afford alkyne N22d (230 mg). 1H NMR (DMSO-d6, 6 = 2.50 ppm, 400 MHz): 6
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12.2/11.9 (br s, 2H), 7.73/7.65 (br s, 2H), 7.62 (app d, 2H), 7.55 (d, J =
2.0, 2H), 7.45
(d, J = 8.0, 2H), 4.65-4.53 (m, 2H), 3.49-3.39 (m, 2H), 2.57-2.44 (obscured,
6H),
2.38-2.19 (m, 4H), 1.68-1.59 (m, 2H), 1.38-1.15 (br s, 18H), 0.80-0.71 (m,
2H), 0.58-
0.52 (m, 2H). LC/MS: Anal. Calcd. for [M+H]+ C42H49N604: 701.37; found 701.4.
Example N22, Step e
Me -71
HNIe" _____________________ (-5 ________ ¨) __ 11--:---(--,1
\ _____________________________ /
4.----- Me
To a solution of alkyne N22d (80 mg, 0.114 mmol) in Me0H (1.5 mL) was
added HC1/dioxane (4M, 5 mL) at 0 C, and the mixture was stirred at room
temperature for 2 h. The volatile component was removed in vacuo, the residue
was
co-evaporated with anhydrous DCM (3 x 5 mL) and dried under high vacuum to
afford pyrrolidine N22e (4HC1, 80 mg) as a pale yellow solid, which was
submitted
to the next step as such. 1H NMR (Me0D, 6 = 3.34 ppm, 400 MHz): 6 8.12 (s,
2H),
7.84 (br s, 2H), 7.74-7.69 (m, 4H), 5.03-4.96 (m, 2H), 3.71-3.62 (m, 2H), 2.90-
2.67
(m, 4H), 2.65 (s, 6H), 2.21-2.14 (m, 2H), 1.12-1.05 (m, 2H), 0.93-0.84 (m,
2H).
LC/MS: Anal. Calcd. for [M+H]+ C32H33N6: 501.27; found 501.2.
Example N22
HATU (88.5 mg, 0.233 mmol) was added to a solution of pyrrolidine N22e
(80 mg, 0.114 mmol), (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (41.8
mg, 0.239 mmol) and DIEA (0.15 mL, 0.91 mmol) in DMF (5 mL). The reaction
mixture was stirred at RT for 2 h. The volatile component was removed in vacuo
and
the residue was dissolved in Et0Ac (50 mL), and washed with saturated NaHCO3
solution and saturated NH4C1, and the combined aqueous phase was back
extracted
with Et0Ac (25 mL). The combined organic extract was washed with water and
brine, dried over Na2504 and concentrated in vacuo. The resulting crude
material
was submitted to a reverse phase HPLC purification (ACN/water/TFA) to afford
Example N22 (40 mg, TFA salt) as an off white solid. LC (Condition 10e and
10e.1):
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>98% homogeneity index. LC/MS (Condition 10e.5):Rt= 1.64 min. 1H NMR
(Me0D, 6 = 3.34 ppm, 400 MHz): 6 7.86 (s, 2H), 7.69 (br s, 2H), 7.66 (d, J =
8.4,
2H), 7.58 (dd, J = 8.4, 1.4, 2H), 5.13/5.11 (d, J = 6.8, 2H), 4.56 (d, J =
6.8, 2H), 3.82-
3.79 (m, 2H), 3.68 (s, 6H), 2.70-2.65 (m, 2H), 2.63 (s, 6H), 2.51-2.43 (m,
2H), 2.20-
2.06 (m, 4H), 1.14-1.08 (m, 2H), 1.02 (d, J = 6.8, 6H), 0.93 (d, J = 6.8, 6H),
0.91-
0.85 (m, 2H). LC/MS: Anal. Calcd. for [M+H]+ C46H55N806: 815.42; found 815.4.
Example N23
0
reir
(\ / ___________________________ :e
0HN---
¨\ Nz,,...." ( 0NH N
0Me
Me0 H N....,7L-N' \¨
,4---- Me\
0
Example N23 (TFA salt) was prepared from pyrrolidine N22e (4HC1 salt) and
(S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid according
to
the procedure described for Example N22. LC (Condition 10e.1 and 10e.4): >94%
homogeneity index. LC/MS (Condition 10e.5):Rt =1.53 min. LC/MS: Anal. Calcd.
for [M+H]+ C50F159N808: 899.44; found 899.4.
Example N24
0
______________________________________________ <H7
Me()
\)_____¨__N N--r N H OMe
---
0
Example N24, Step a
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N2---------y-.--yr----N
0 Boc
To a solution of (1R,3S,5R)-2-(tert-butoxycarbony1)-2-
azabicyclo[3.1.0]hexane-3-carboxylic acid (4.52 g, 20.0 mmol) in THF (60 mL),
cooled to -25 C, TEA (2.78 mL, 20.0 mmol) followed by isobutyl chloroformate
(2.58 mL, 20.0 mmol) were added. After 30 minutes of stirring, the reaction
mixture
was warmed to -10 C and CH2N2 in Et20 (3.0 eq ) was added and stirred at the
same
temperature for 3 h and at room temperature for 12 h. The volatile components
were
evaporated under reduced pressure and the crude was purified by flash
chromatography (ISCO, Silica gel; Et0Ac: petroleum ether, 30:70) to afford
N24a
(3.9 g) as a yellow solid. 1H NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 5.50 (s,
1H),
4.01 (br s, 1H), 3.35 (br s, 1H), 2.27 (br s, 2H), 1.60-1.56 (m, 1H), 1.45 (s,
9H), 0.83-
0.80 (m, 1H), 0.44 (br s, 1H).
Example N24, Step b
f-9r
Br----- N
0 Boc
Aqueous HBr (48%, 2.0 mL) was added to a stirred solution of N24a (2.0 g,
7.96 mmol) in Et20 (20 mL) at 0 C. After 5 minutes of stirring, the reaction
mixture
was diluted with Et0Ac (100 mL) and the organic phase was washed with water
(50
mL) and brine (25 mL), dried over Na2504 and concentrated in vacuo to afford
crude
N24b (2.02 g) as a pale yellow liquid which was used as such in the next step.
1H
NMR (CDC13, 6 = 7.26 ppm, 400 MHz): 6 4.21-4.14 (m, 1H), 4.10 (s, 2H),
3.52/3.40
(m, 1H), 2.41-2.17 (m, 2H), 1.68-1.61 (m, 1H), 1.47 (s, 9H), 0.85-0.78 (m,
1H), 0.52
(br s, 1H).
Example N24, Step c
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.-- N =
Boc I \ Br
,k_...... H
To a stirred solution of 4-bromobenzimidamide (384 mg, 1.65 mmol) in
THF:H20 (10 mL: 2.5 mL), K2CO3 (456 mg, 3.30 mmol) was added followed by
N24b (500 mg, 1.65 mmol) in THF (10 mL) and stirred at RT for 24 h. The
volatile
component was removed under reduced pressure and the crude was dissolved in
Et0Ac (100 mL) and the organic phase was washed with water (50 mL) and brine
(25 mL), dried over Na2SO4 and concentrated in vacuo to afford a mixture of
the N-
alkylated precursor and targeted product. To this mixture (1.65 mmol) in
xylenes (8
mL) was added NH40Ac (1.27 g, 16.5 mmol), and the reaction mixture was
transferred into a pressure tube, purged with N2 for 10 minutes and heated at
130 C
for overnight. The volatile component was removed under reduced pressure, the
crude was treated with saturated NaHCO3 solution and extracted with Et0Ac (100
mL). The organic layer was washed with water (50 mL) and brine (25 mL), dried
over Na2SO4 and concentrated in vacuo. The resulting crude material was
purified
by flash chromatography (Silica gel; MeOH:DCM: 2:98) to afford imidazole N24c
(350 mg) as a pale yellow solid. 1H NMR (Me0D, 6 = 3.34 ppm, 400 MHz): 6 7.79-
7.76 (m, 2H), 7.63-7.61 (m, 2H), 6.96 (s, 1H), 4.76 (app t, 1H), 3.49 (app t,
1H),
2.44-2.32 (m, 2H), 1.71-1.64 (m, 1H), 1.38 (s, 9H), 0.92-0.87 (m, 1H), 0.58-
0.52 (m,
1H). LC/MS: Anal. Calcd. for [M+H]+ C19H23BrN302: 404.09; found 405Ø
Example N24, Step d
_______________________________________________ H -71
---1 ¨/)_____(¨)__<\N -.....7-- N
I Boc
Boc
4_, H
A solution of N24c (135 mg, 0.33 mmol) in DMF (2 mL) was purged with N2
for 10 minutes, Pd(Ph3P)4 (14 mg, 0.012 mmol), bis-trimethylstanylacetylene
(58 mg,
0.16 mmol) were added, and the reaction mixture was purged with N2 for
additional
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minutes and heated with a microwave at 90 C for 3 h. The reaction mixture was
treated with Et0Ac (50 mL) and washed with water (25 mL) and brine (10 mL),
dried over Na2SO4 and concentrated in vacuo. The resulting crude was purified
by
flash chromatography (ISCO, basic A1203; Et0Ac: petroleum ether, 15:85) to
afford
5 alkyne N24d (14 mg) as a pale yellow solid. 1H NMR (Me0D, 6 = 3.34 ppm,
400
MHz): 6 7.90 (d, J = 8.8, 4H), 7.63 (d, J = 8.8, 4H), 6.99 (s, 2H), 4.79 (br
s, 2H), 3.50
(br s, 2H), 2.44-2.35 (m, 4H), 1.73-1.69 (m, 2H), 1.47 (s, 18H), 0.94-0.86 (m,
2H),
0.59 (br s, 2H). LC/MS: Anal. Calcd. for [M+H]+ C40I-145N604: 673.34; found
673.4.
10 Example N24, Step e
¨ _____________________________________________ H1
¨%
,44....õ...i H
To a solution of N24d (14 mg, 0.020 mmol) in Me0H (0.5 mL) was added
HC1/dioxane (4M, 1 mL) at 0 C, and the mixture was stirred at room
temperature for
2 h. The volatile component was removed in vacuo, the residue was co-
evaporated
with anhydrous DCM (3 x 5 mL) and dried under high vacuum to afford
pyrrolidine
N24e (4HC1, 12.8 mg) as a pale yellow solid, which was submitted to the next
step as
such. 1H NMR (Me0D, 6 = 3.34 ppm, 400 MHz): 6 8.08 (d, J = 8.4, 4H), 7.88 (s,
2H), 7.86 (d, J = 8.4, 4H), 4.78 (obscured, 2H), 3.54 (app t, 2H), 2.73-2.66
(m, 4H),
2.12-2.09 (m, 2H), 1.08-1.02 (m, 2H), 0.92-0.81 (m, 2H). LC/MS: Anal. Calcd.
for
[M+H]+ C30I-129N6: 473.24; found 473.2.
Example N24
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0
C H7
Me0
0 ,r \ j/ )____(¨/) (\N-1,---)N L/OMe
1
H N
A146._ H
0
HATU (16 mg, 0.042 mmol) was added to a solution of N24e (12.8 mg, 0.020
mmol), (S)-2-(methoxycarbonylamino)-2-(tetrahydro-2H-pyran-4-yl)acetic acid (9
mg, 0.04 mmol) and DIEA (0.014 mL, 0.08 mmol) in DMF (2 mL). After stirring
for
2 h at RT, the reaction mixture was dissolved in Et0Ac (50 mL), and washed
with
saturated NH4C1 (25 mL), saturated NaHCO3 solution (25 mL), brine (10 mL),
dried
over Na2SO4 and concentrated in vacuo. The crude was submitted to a reverse
phase
HPLC purification (ACN/water/TFA) to afford Example N24 (8 mg, TFA salt) as
white solid. LC (Condition 10e.2 and 10e.8): >92% homogeneity index. LC/MS
(Condition 10e.6):Rt= 1.46 min. 1H NMR (Me0D, 6 = 3.34 ppm, 400 MHz): 6 7.94
(d, J = 8.4, 4H), 7.83 (d, J = 8.4, 4H), 7.52 (s, 2H), 5.25-5.20 (m, 2H), 4.64
(d, J =
7.6, 2H), 3.99-3.90 (m, 4H), 3.74-3.69 (m, 2H), 3.69 (s, 6H), 3.42-3.30 (m,
4H),
2.52-2.43 (m, 4H), 2.07-2.02 (m, 4H), 1.64-1.31 (m, 8H), 1.18-1.16 (m, 2H),
0.88-
0.83 (m, 2H). LC/MS: Anal. Calcd. for [M+H]+ C48H55N808: 871.41; found 871.4.
Example N25
H
'NH/ )(¨/)
Me0 H N---../ 'NI
.414._ H
Example N25 (free base) was prepared similarly from pyrrolidine N24e and
(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid according to the procedure
described for Example N24, with the notable exception that CH3CN/Water/NH40Ac
was used for the reverse phase HPLC purification step. LC (Condition 10e and
10e.9): >96% homogeneity index. LC/MS (Condition 10e.6):Rt=1.67 min. LC/MS:
Anal. Calcd. for [M+H]+ C44H51N806: 787.39; found 787.4.
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Example J1 to J1.1
0
410 V N
NH
HN¨)4
0 N '
Example J1 0
0
>\--NH 4111 V N
NH
HN
4s1H
0 N 404
Example J1.1
Example J1, step a
Br
I 0
00
DIPEA (2.8 mL, 16 mmol) was added to a slurry of acid M3f (2.00 g, 8.80
mmol) and 2-bromo-1-(3-bromophenyl)ethanone (2.22 g, 8.00 mmol) in
acetonitrile
(25 mL) and the reaction mixture was stirred overnight at rt. The reaction
mixture
was concentrated and purified with a Biotage Horizon (80g Si02, 10-25%
Et0Ac/hexanes) to yield ketoester Jla (3.37 g) as a viscous light yellow oil.
LC-MS
retention time 1.853 min; m/z 423 and 425.98 (1:1) (MH+). LC data was recorded
on a Shimadzu LC-10AS liquid chromatograph equipped with a Waters Xterra MS
7u C18 3.0x5Omm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220nM. The elution conditions employed a flow rate of 4 mL/min , a
gradient of 100% solvent A / 0% solvent B to 0% solvent A / 100% solvent B, a
gradient time of 2 min, a hold time of 1 min, and an analysis time of 3 min
where
solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate and solvent B was
5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode. 1H NMR (400 MHz, CDC13) 6
ppm 8.04 (t, J=1.8 Hz, 1H), 7.83 (d, J=7.8 Hz, 1H), 7.75 (d, J=8.0 Hz, 1H),
7.38 (t,
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J=7.9 Hz, 1H), 5.33 - 5.59 (m, 1H), 5.13 - 5.32 (m, 1H), 4.22 (br s, 1H), 3.40
- 3.63
(m, 1H), 2.52 - 2.63 (m, 1H), 2.46 (dd, J=13.1, 9.3 Hz, 1H), 1.63 - 1.73 (m,
1H), 1.47
(br s, 9H), 0.86 (br s, 1H), 0.51 (br s, 1H).
Example J1, step b
-/b.
0"--A-NH
Br
)c 0 N 7 =
A solution of ketoester Jla (3.33 g, 7.85 mmol) in xylene (75 mL) was added
to ammonium acetate (5.94 g, 77 mmol) in a pressure vessel and the reaction
mixture
was stirred under nitrogen for 5 min. The reaction vessel was sealed and then
placed
into an oil bath which had been preheated to 140 C and the reaction was held
at that
temperature for 6h. The reaction was cooled to rt, stirred overnight and
reheated at
140 C for 5 hr. Additional ammonium acetate (3.0g) was added and the reaction
was stirred at 145 C for 8 hrs, cooled to rt and concentrated under high
vacuum to a
brown oil. The oil was partitioned between DCM (¨ 200 mL) and 1/2 sat. sodium
bicarbonate (-200 mL). The organic layer was dried (MgSO4), filtered and
concentrated. The crude orange solidified foam was purified on a Biotage
Horizon
(20-50% Et0Ac/hexanes, 160 g Si02) to yield imidazole Jib (2.03 g) as a yellow
solidified foam. LC-MS retention time 2.450 min; m/z 404 and 406.06 (1:1) (MH-
0.
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph equipped with
a Phenomenex-Luna 10u C18 3.0x5Omm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220 nM. The elution conditions employed
a
flow rate of 4 mL/min , a gradient of 100% solvent A / 0% solvent B to 0%
solvent A
/ 100% solvent B, a gradient time of 3 min, a hold time of 1 min, and an
analysis time
of 4 min where solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate
and solvent B was 5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray mode. 1H NMR (400
MHz, Me0D) 6 ppm 7.89 (br s, 1H), 7.65 (d, J=6.5 Hz, 1H), 7.32 - 7.44 (m, 2H),
7.26 (t, J=7.8 Hz, 1H), 4.66 (br s, 1H), 3.52 - 3.63 (m, 1H), 2.51 (dd,
J=13.1, 8.8 Hz,
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1H), 2.25 - 2.37 (m, 1H), 1.66 - 1.75 (m, 1H), 1.29 (br s, 9H), 0.84 (ddd,
J=8.2, 6.0,
5.8 Hz, 1H), 0.56 - 0.63 (m, 1H).
Example J1, step c
40 V N 0 \I____
0"--A-NH
HN -1V----- 0
Nitrogen was bubbled through a stirred solution of bromide Jib (854 mg, 2.18
mmol) and 1,2-bis(trimethylstannyl)ethyne (403 mg, 1.15 mmol) in DMF (10 mL)
for 10 minutes. Then Pd(PPh3)4 (79 mg, 0.069 mmol) was added, nitrogen was
bubbled through the reaction for 1 min and then the reaction was heated at 90
C
under nitrogen for 17h. The reaction mixture was cooled to rt, concentrated to
a
viscous oil and purified on a Biotage Horizon (40 g Si02, 70-100%
Et0Ac/hexanes,
loaded with DCM)) to yield alkyne J lc (500 mg) as a yellow solidified foam.
LC-MS retention time 2.876 min; m/z 649.51 (MH+). LC data was recorded on a
Shimadzu LC-10AS liquid chromatograph equipped with a Phenomenex-Luna 10u
C18 3.0x5Omm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 4 mL/min , a
gradient of 100% solvent A / 0% solvent B to 0% solvent A / 100% solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time of 4 min
where
solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate and solvent B was
5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode. 1H NMR (400 MHz, Me0D) 6
ppm 7.89 (s, 2H), 7.67 - 7.74 (m, 2H), 7.35 - 7.44 (m, 6H), 4.85 - 5.04 (m,
2H), 3.64 -
3.74 (m, 2H), 3.47 - 3.56 (m, 2H), 2.29 - 2.45 (m, 2H), 1.90 - 2.13 (m, 6H),
1.48 (br
s, 6H), 1.26 (br s, 12H).
Example J1, step d
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HS- . V N
NH HN-vi)
/
N v = NH
TFA (3 mL, 38.9 mmol) was added to a stirred solution of carbamate He
(238 mg, 0.367 mmol) in DCE (7 mL) and the reaction was stirred at rt for lh.
The
reaction mixture was concentrated under vacuum to yield a TFA salt of
pyrrolidine
Jld (260 mg) as a yellow solid.
LC-MS retention time 2.505 min; m/z 449.22 (MH+). LC data was recorded
on a Shimadzu LC-10AS liquid chromatograph equipped with a Phenomenex-Luna
10u C18 3.0x5Omm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 4 mL/min , a
gradient of 100% solvent A / 0% solvent B to 0% solvent A / 100% solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time of 4 min
where
solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate and solvent B was
5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode. 1H NMR (400 MHz, Me0D) 6
ppm 7.96 - 8.00 (m, 2H), 7.79 (dt, J=6.6, 2.1 Hz, 2H), 7.65 (s, 2H), 7.41 -
7.48 (m,
4H), 4.91 (t, J=7.8 Hz, 2H), 3.44 - 3.61 (m, 4H), 2.51 -2.62 (m, 2H), 2.27 -
2.45 (m,
4H), 2.16 - 2.24 (m, 2H).
Example J1 to J1.1
HATU (61.8 mg, 0.162 mmol) was added to a stirred solution of a TFA salt of
pyrrolidine Jld (110 mg, 0.12 mmol) and (S)-2-(methoxycarbonylamino)-3-
methylbutanoic acid (28.5 mg, 0.162 mmol) in DMF (0.7 mL) and Hunig'sBase
(0.10
mL, 0.54 mmol). The reaction mixture was stirred at rt for 3 hours and then
concentrated under a stream of nitrogen. The reaction was purified in two
injections
by preparative HPLC (Me0H/water /10 mM ammonium acetate) to yield Example J1
(31.5 mg) as a light yellow solid and Example J1.1 (41.1 mg) as a a light
yellow
solid.
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Example J1: LC-MS retention time 2.605 min; m/z 763.36 (MH+). LC data
was recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 3.0x5Omm column using a SPD-10AV UV-Vis detector
at a detector wave length of 220 nM. The elution conditions employed a flow
rate of
4 mL/min, a gradient of 100% solvent A / 0% solvent B to 0% solvent A / 100%
solvent B, a gradient time of 3 min, a hold time of 1 min, and an analysis
time of 4
min where solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate and
solvent B was 5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray mode. The 1H NMR
presents as a 4 :1 mixture of rotamers. The major rotamer is: 1H NMR (400 MHz,
Me0D) 6 ppm 7.86 (s, 2H), 7.64 - 7.70 (m, 2H), 7.36 - 7.43 (m, 4H), 7.34 (s,
2H),
5.17 (dd, J=7.8, 5.3 Hz, 2H), 4.24 (d, J=7.3 Hz, 2H), 3.95 - 4.04 (m, 2H),
3.84 - 3.92
(m, 2H), 3.66 (s, 6H), 2.17 - 2.42 (m, 6H), 2.00 - 2.13 (m, 4H), 0.96 (d,
J=6.8 Hz,
6H), 0.91 (d, J=6.8 Hz, 6H).
Example J1.1: LC-MS retention time 2.620 min; m/z 606.23 (MH+). LC
data was recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 3.0x5Omm column using a SPD-10AV UV-Vis detector
at a detector wave length of 220 nM. The elution conditions employed a flow
rate of
4 mL/min , a gradient of 100% solvent A / 0% solvent B to 0% solvent A / 100%
solvent B, a gradient time of 3 min, a hold time of 1 min, and an analysis
time of 4
min where solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate and
solvent B was 5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray mode. The 1H NMR
presents as a 3 :1 mixture of rotamers. 1H NMR (400 MHz, Me0D) 6 ppm 7.94 (s,
1H), 7.87 (s, 1H), 7.74 (ddd, J=5.6, 3.4, 1.8 Hz, 1H), 7.65 - 7.70 (m, 1H),
7.51 (s,
1H), 7.36 - 7.44 (m, 6H), 7.35 (s, 1H), 5.18 (dd, J=7 .7 , 5.4 Hz, 1H), 4.68
(t, J=7.8
Hz, 1H), 4.24 (d, J=7.5 Hz, 1H), 3.95 - 4.06 (m, 1H), 3.85 - 3.92 (m, 1H),
3.66 (s,
3H), 3.35 - 3.52 (m, 2H), 2.01 - 2.50 (m, 9H), 0.96 (d, J=6.8 Hz, 3H), 0.91
(d, J=6.8
Hz, 3H).
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Example J2
0
0
----0 - N 0
NH
/ 0 HN-i)
N v ilo N ?"---
0
0
HATU (118 mg, 0.310 mmol) was added to a stirred solution of a TFA salt of
pyrrolidine Jld (93.4 mg, 0.103 mmol) and (S)-2-(methoxycarbonylamino)-2-
(tetrahydro-2H-pyran-4-yl)acetic acid (67.3 mg, 0.310 mmol) in DIPEA (0.18 mL,
1.0 mmol) and DMF (1 mL) and the reaction was stirred at rt for 2h. The
reaction
was concentrated under a stream of nitrogen, dissolved into Me0H and purified
by
preparative HPLC (Me0H/water 10 mM ammonium acetate) to yield Example J2
(18.1 mg) as a light pink solid
LC-MS retention time 2.428 min; m/z 847.39 (MH+). LC data was recorded
on a Shimadzu LC-10AS liquid chromatograph equipped with a Phenomenex-Luna
10u C18 3.0x5Omm column using a SPD-10AV UV-Vis detector at a detector wave
length of 220 nM. The elution conditions employed a flow rate of 4 mL/min , a
gradient of 100% solvent A / 0% solvent B to 0% solvent A / 100% solvent B, a
gradient time of 3 min, a hold time of 1 min, and an analysis time of 4 min
where
solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate and solvent B was
5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was determined using a
Micromass Platform for LC in electrospray mode. The 1H NMR exhibits as a 4:1
mixture of rotamers. The major rotamer is : 1H NMR (400 MHz, Me0D) 6 ppm
7.83 (s, 2H), 7.62 - 7.67 (m, 2H), 7.33 - 7.42 (m, 6H), 5.15 (dd, J=7.5, 5.5
Hz, 2H),
4.32 (d, J=8.3 Hz, 2H), 3.89 - 4.08 (m, 6H), 3.66 (s, 6H), 3.33 - 3.50 (m,
6H), 2.23 -
2.40 (m, 4H), 2.13 -2.22 (m, 2H), 1.92 -2.10 (m, 4H), 1.61 - 1.71 (m, 4H),
1.32 -
1.56 (m, 4H).
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Example J3
0 k_isi
---.0 -........?__i . V N 00_____
NH HN¨µ.) 1
ii...N._.\(
- 0
'/-
Example J3, step a
4.:z.,.....
H Br
)C 0
Bromide J3a was prepared from acid M3f according to the procedure
described for bromide Jib.
Example J3, step b
>K
" ¨
NH
HNA),,-0 O Ni 7
=of
Nitrogen was bubbled through a stirred solution of bromidie J3a (825 mg,
2.041 mmol) and 1,2-bis(trimethylstannyl)ethyne (378 mg, 1.074 mmol) in DMF
(10
mL) for 10 minutes. Then, Pd(PPh3)4 (74.5 mg, 0.064 mmol) was added to the
reaction mixture, nitrogen was bubbled through the reaction for 1 min and then
the
reaction was heated at 90 C under nitrogen for 17h. The reaction mixture was
cooled to rt, concentrated to a thick black oil, diluted with DCM and purified
on a
Biotage Horizon (40 g Si02, 60-100% Et0Ac/hexanes) to yield alkyne J3b (312
mg)
as a yellow solid. LC-MS retention time 2.800 min; m/z 671.53 (MH-). LC data
was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 3.0x5Omm column using a SPD-10AV UV-Vis detector
at a detector wave length of 220 nM. The elution conditions employed a flow
rate of
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4 mL/min , a gradient of 100% solvent A / 0% solvent B to 0% solvent A / 100%
solvent B, a gradient time of 3 min, a hold time of 1 min, and an analysis
time of 4
mM where solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate and
solvent B was 5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray mode. 1H NMR (400
MHz, Me0D) 6 ppm 7.89 (s, 2H), 7.70 (ddd, J=5.6, 3.3, 1.9 Hz, 2H), 7.38 - 7.43
(m,
6H), 4.69 (br s, 2H), 3.59 (br s, 2H), 2.53 (dd, J=13.2, 8.9 Hz, 2H), 2.28 -
2.39 (m,
2H), 1.68 - 1.77 (m, 2H), 1.29 - 1.31 (m, 18H), 0.85 (dt, J=8.5, 5.8 Hz, 2H),
0.58 -
0.64 (m, 2H).
Example J3, step c
-ih.
HS¨ 411 V N
NH
/ HN
-)1H
N7 .
,, --
/7.-
TFA (3 mL, 38.9 mmol) was added to a stirred solution of carbamate J3b
(316 mg, 0.470 mmol) in DCE (7 mL) and the reaction was stirred at rt for lh.
The
Example J3
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HATU (69.6 mg, 0.183 mmol) was added to a stirred solution of a TFA salt of
pyrrolidine J3c (56.7 mg, 0.061 mmol) and (S)-2-(methoxycarbonylamino)-3-
methylbutanoic acid (32.1 mg, 0.183 mmol) in DMF (0.7 mL) and Hunig'sBase
(0.11
mL, 0.61 mmol) and the reaction mixture was stirred at rt for 3 h. The
reaction was
concentrated under a stream of nitrogen, dissolved into Me0H and purified in
two
injections by preparative HPLC (Me0H/water 10 mM ammonium acetate) to yield
Example J3 (31.8 mg) as a light yellow solid. LC-MS retention time 2.605 min;
m/z
787.36 (MH+). LC data was recorded on a Shimadzu LC-10AS liquid
chromatograph equipped with a Phenomenex-Luna 10u C18 3.0x5Omm column using
a SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The elution
conditions employed a flow rate of 4 mL/min , a gradient of 100% solvent A /
0%
solvent B to 0% solvent A / 100% solvent B, a gradient time of 3 min, a hold
time of
1 min, and an analysis time of 4 min where solvent A was 5% Me0H / 95% H20 /
10
mM ammonium acetate and solvent B was 5% H20 / 95% Me0H / 10 mM
ammonium acetate. MS data was determined using a Micromass Platform for LC in
electrospray mode. 1H NMR (400 MHz, Me0D) 6 ppm 7.87 (s, 2H), 7.65 - 7.71 (m,
2H), 7.37 - 7.43 (m, 4H), 7.36 (s, 2H), 5.16 (dd, J=8.8, 4.8 Hz, 2H), 4.59 (d,
J=6.5
Hz, 2H), 3.67 (s, 6H), 3.63 - 3.68 (m, 2H), 2.49 - 2.58 (m, 2H), 2.37 - 2.45
(m, 2H),
2.10 -2.21 (m, 2H), 1.98 - 2.07 (m, 2H), 1.12 (ddd, J=8.7, 5.6, 5.5 Hz, 2H),
1.01 (d,
J=6.8 Hz, 6H), 0.93 (d, J=6.8 Hz, 6H), 0.78 (br s, 2H).
Example J4
0
0
ZN 0
N
= - HN - 0--
0 N 404
0
.
0
HATU (120 mg, 0.317 mmol) was added to a stirred solution of a TFA salt of
pyrrolidine J3c (98 mg, 0.106 mmol) and (S)-2-(methoxycarbonylamino)-2-
(tetrahydro-2H-pyran-4-yl)acetic acid (68.8 mg, 0.317 mmol) in DIPEA (0.18 mL,
1.1 mmol) and DMF (1 mL) and the reaction was stirred at rt for 3h. The
reaction
was concentrated under a stream of nitrogen, dissolved into Me0H and purified
in by
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preparative HPLC (Me0H/water 10 mM ammonium acetate) and then repurified by
prep HPLC (Me0H/water 0.1% TFA) to yield a TFA salt of Example J4 (18 mg) as a
white solid. LC-MS retention time 2.463 min; m/z 869.40 (MH-). LC data was
recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a
Phenomenex-Luna 10u C18 3.0x5Omm column using a SPD-10AV UV-Vis detector
at a detector wave length of 220 nM. The elution conditions employed a flow
rate of
4 mL/min, a gradient of 100% solvent A / 0% solvent B to 0% solvent A / 100%
solvent B, a gradient time of 3 min, a hold time of 1 min, and an analysis
time of 4
min where solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate and
solvent B was 5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray mode. 1H NMR (400
MHz, Me0D) 6 ppm 7.89 - 7.92 (m, 2H), 7.87 (s, 2H), 7.74 (dt, J=7.8, 1.5 Hz,
2H),
7.61 -7.66 (m, 2H), 7.54 - 7.60 (m, 2H), 5.11 (dd, J=9.0, 7.0 Hz, 2H), 4.59
(d, J=7.5
Hz, 2H), 3.91 - 3.99 (m, 4H), 3.83 (t, J=4.8 Hz, 2H), 3.67 (s, 6H), 3.33 -
3.45 (m,
4H), 2.67 (dd, J=13.7, 9.4 Hz, 2H), 2.43 - 2.52 (m, 2H), 2.02 - 2.14 (m, 4H),
1.39 -
1.63 (m, 8H), 1.03 - 1.11 (m, 2H), 0.83 - 0.90 (m, 2H).
Example J5
NH
0 0
Hs,\FL. TH.
0 ErN
Example J5, Step a
N
Nitrogen was bubbled through a stirred solution of (5)-tert-butyl 24444-
bromopheny1)-1H-imidazol-2-y1)pyrrolidine-1-carboxylate (bromide D- lb) (400
mg,
1.02 mmol), 4-ethynylphenylboronic acid (149 mg, 1.02 mmol) and sodium
bicarbonate (214 mg, 2.55 mmol) in DME (8 mL) and water (2 mL) for 15 min.
Then Pd(PPh3)4 (58.9 mg, 0.051 mmol) was added and the nitrogen bubbling was
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continued for 5 min. before the reaction vessel was sealed and heated at 90 C
overnight. The reaction was cooled, diluted with THF, washed with brine and
concentrated to dryness. The residue was purified by Biotage Horizon (60-90%
Et0Ac/hexanes) to yield carbamate J5a (85 mg) as a yellow solid. LC-MS
retention
time 2.911 min; m/z 723.64 (MH-). LC data was recorded on a Shimadzu LC-10AS
liquid chromatograph equipped with a Phenomenex-Luna 10u C18 3.0x5Omm
column using a SPD-10AV UV-Vis detector at a detector wave length of 220nM.
The elution conditions employed a flow rate of 4 mL/min , a gradient of 100%
solvent A / 0% solvent B to 0% solvent A / 100% solvent B, a gradient time of
3 min,
a hold time of 1 min, and an analysis time of 4 min where solvent A was 5%
Me0H /
95% H20 / 10 mM ammonium acetate and solvent B was 5% H20 / 95% Me0H / 10
mM ammonium acetate. MS data was determined using a Micromass Platform for
LC in electrospray mode. 1H NMR (400 MHz, Me0D) 6 ppm 7.78 (d, J=8.0 Hz,
2H), 7.72 (d, J=8.0 Hz, 2H), 7.64 - 7.69 (m, 4H), 7.58 (d, J=8.3 Hz, 2H), 7.52
(d,
J=8.3 Hz, 2H), 7.31 - 7.41 (m, 2H), 4.86 - 5.03 (m, 2H), 3.68 (br s, 2H), 3.46
-
3.57 (m, 2H), 2.37 (br s, 2H), 1.89 -2.13 (m, 6H), 1.47 (br s, 6H), 1.25 (br
s, 12H).
Example J5, Step b
H _
ss\L"z'N - -
c---
TFA (250 L, 3.24 mmol) was added to a solution of carbamate J5a in DCE
(1 mL) and the reaction was stirred at rt for 1.5 hr. The reaction was
concentrated
under a stream of nitrogen to provide a TFA salt of pyrrolidine J5b (45 mg).
LC-MS
retention time 2.928 min; m/z 525.32 (MH+). LC data was recorded on a Shimadzu
LC-10AS liquid chromatograph equipped with a Phenomenex-Luna 10u C18 3.0 x
50mm column using a SPD-10AV UV-Vis detector at a detector wave length of
220nM. The elution conditions employed a flow rate of 4 mL/min, a gradient of
100% solvent A / 0% solvent B to 0% solvent A / 100% solvent B, a gradient
time of
4 min, a hold time of 1 min, and an analysis time of 5 min where solvent A was
5%
Me0H / 95% H20 / 10 mM ammonium acetate and solvent B was 5% H20 / 95%
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Me0H / 10 mM ammonium acetate. MS data was determined using a Micromass
Platform for LC in electrospray mode.
Example J5
HATU (53 mg, 0.14 mmol) was added to a solution of a TFA salt of J5b (45
mg, 0.046 mmol) and (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (24.2
mg, 0.138 mmol) in DMF (0.7 mL) and TEA (0.038 mL, 0.28 mmol) and the reaction
was stirred for 30 min. The reaction was diluted with Me0H, filtered and
purified by
prep HPLC (Me0H/water with 10 mM ammonium acetate) to yield Example J5 (14.7
mg) as a light yellow solid. LC-MS retention time 2.728 min; m/z 839.57 (MH-0.
LC data was recorded on a Shimadzu LC-10AS liquid chromatograph equipped with
a Phenomenex-Luna 10u C18 3.0x5Omm column using a SPD-10AV UV-Vis
detector at a detector wave length of 220nM. The elution conditions employed a
flow rate of 4 mL/min , a gradient of 100% solvent A / 0% solvent B to 0%
solvent A
/ 100% solvent B, a gradient time of 4 min, a hold time of 1 min, and an
analysis time
of 5 min where solvent A was 5% Me0H / 95% H20 / 10 mM ammonium acetate
and solvent B was 5% H20 / 95% Me0H / 10 mM ammonium acetate. MS data was
determined using a Micromass Platform for LC in electrospray mode. 1H NMR (400
MHz, Me0D) 6 ppm 7.73 - 7.78 (m, 2H), 7.65 - 7.71 (m, 6H), 7.57 - 7.62 (m,
2H),
7.52 (d, J=8.5 Hz, 2H), 7.33 - 7.36 (m, 2H), 5.18 (dt, J=7.6, 5.2 Hz, 2H),
4.21 -
4.27 (m, 2H), 3.96 - 4.05 (m, 2H), 3.84 - 3.92 (m, 2H), 3.66 (s, 6H), 2.17 -
2.42 (m,
6H), 2.00 - 2.13 (m, 4H), 0.95 (dd, J=6.8, 1.3 Hz, 6H), 0.91 (dd, J=6.8, 1.0
Hz,
6H).
Synthesis of common caps
Compound analysis conditions: Purity assessment and low resolution mass
analysis were conducted on a Shimadzu LC system coupled with Waters Micromass
ZQ MS system. It should be noted that retention times may vary slightly
between
machines. Additional LC conditions applicable to the current section, unless
noted
otherwise.
Cond.-MS-WI
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Column = XTERRA 3.0 X 50 mm S7
Start %B =0
Final %B = 100
Gradient time = 2 min
Stop time = 3 min
Flow Rate =5 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Cond.-MS-W2
Column = XTERRA 3.0 X 50 mm S7
Start %B =0
Final %B = 100
Gradient time = 3 min
Stop time = 4 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Cond.-MS-W5
Column = XTERRA 3.0 X 50 mm S7
Start %B =0
Final %B =30
Gradient time = 2 min
Stop time = 3 min
Flow Rate =5 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
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Cond.-D1
Column = XTERRA C18 3.0 X 50 mm S7
Start %B =0
Final %B = 100
Gradient time = 3 min
Stop time = 4 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Cond.-D2
Column = Phenomenex-Luna 4.6 X 50 mm S10
Start %B =0
Final %B =100
Gradient time = 3 min
Stop time = 4 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Cond.-MD]
Column = XTERRA 4.6 X 50 mm S5
Start %B =0
Final %B = 100
Gradient time = 3 min
Stop time = 4 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
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Cond.-M3
Column = XTERRA C18 3.0 X 50 mm S7
Start %B =0
Final %B = 40
Gradient time = 2 min
Stop time = 3 min
Flow Rate =5 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Condition OL1
Column = Phenomenex-Luna 3.0 X 50 mm S10
Start %B =0
Final %B = 100
Gradient time = 4 min
Stop time = 5 min
Flow Rate =4 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Condition 0L2
Column = Phenomenex-Luna 50X 2 mm 3 u
Start %B = 0
Final %B = 100
Gradient time = 4 min
Stop time = 5 min
Flow Rate = 0.8 mL/min
Oven Temp = 40 C
Wavelength = 220 nm
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Solvent A = 0.1% TFA in 10% Acetonitrile/90%H20
Solvent B = 0.1% TFA in 90% Acetonitrile/10% H20
Condition I
Column = Phenomenex-Luna 3.0 X 50 mm S10
Start %B =0
Final %B = 100
Gradient time = 2 min
Stop time = 3 min
Flow Rate = 4 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Condition II
Column = Phenomenex-Luna 4.6 X 50 mm S10
Start %B =0
Final %B = 100
Gradient time = 2 min
Stop time = 3 min
Flow Rate = 5 mL/min
Wavelength = 220 nm
Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Condition III
Column = XTERRA C18 3.0 x 50mm S7
Start %B =0
Final %B = 100
Gradient time = 3 min
Stop time = 4 min
Flow Rate =4 mL/min
Wavelength = 220 nm
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Solvent A = 0.1% TFA in 10% methanol/90%H20
Solvent B = 0.1% TFA in 90% methanol/10% H20
Cap-1
So
. OH
n
A suspension of 10% Pd/C (2.0g) in methanol (10 mL) was added to a
mixture of (R)-2-phenylglycine (10g, 66.2 mmol), formaldehyde (33 mL of 37%
wt.
in water), 1N HC1 (30 mL) and methanol (30 mL), and exposed to H2 (60 psi) for
3
hours. The reaction mixture was filtered through diatomaceous earth
(Celite()), and
the filtrate was concentrated in vacuo. The resulting crude material was
recrystallized from isopropanol to provide the HC1 salt of Cap-1 as a white
needle
(4.0 g). Optical rotation: -117.1 [c=9.95 mg/mL in H20; 2=589 nm]. 1H NMR
(DMSO-d6, 6=2.5 ppm, 500 MHz): 6 7.43-7.34 (m, 5H), 4.14 (s, 1H), 2.43 (s,
6H);
LC (Cond. I): RT=0.25; LC/MS: Anal. Calcd. for [M+H]+ C10H14NO2 180.10;
found 180.17; HRMS: Anal. Calcd. for [M+H]+ C10H14NO2 180.1025; found
180.1017.
Cap-2
SO
. OH
n
r 1
NaBH3CN (6.22g, 94 mmol) was added in portions over a few minutes to a
cooled (ice/water) mixture of (R)-2-Phenylglycine (6.02 g, 39.8 mmol) and
methanol
(100 mL), and stirred for 5 minutes. Acetaldehyde (10 mL) was added dropwise
over
10 minutes and stirring was continued at the same cooled temperature for 45
minutes
and at ambient temperature for ¨6.5 hours. The reaction mixture was cooled
back
with ice-water bath, treated with water (3 mL) and then quenched with a
dropwise
addition of concentrated HC1 over ¨ 45 minutes until the pH of the mixture was
¨ 1.5
¨ 2Ø The cooling bath was removed and the stirring was continued while
adding
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concentrated HC1 in order to maintain the pH of the mixture around 1.5-2Ø
The
reaction mixture was stirred overnight, filtered to remove the white
suspension, and
the filtrate was concentrated in vacuo. The crude material was recrystallized
from
ethanol to afford the HC1 salt of Cap-2 as a shining white solid in two crops
(crop-1:
4.16 g; crop-2: 2.19 g). 1H NMR (DMSO-d6, cF2.5 ppm, 400 MHz): 10.44 (1.00, br
s, 1H), 7.66 (m, 2H), 7.51 (m, 3H), 5.30 (s, 1H), 3.15 (br m, 2H), 2.98 (br m,
2H),
1.20 (app br s, 6H). Crop-1: [a]25 -102.21 (c=0.357, H20); crop-2: [a]25 -
99.7
(c=0.357, H20). LC (Cond. I): RT=0.43 min; LC/MS: Anal. Calcd. for [M+H]+
C12H18NO2: 208.13; found 208.26.
Cap-3
so
. OH
N
1
Acetaldehyde (5.0 mL, 89.1 mmol) and a suspension of 10% Pd/C (720 mg)
in methanol/H20 (4mL/1 mL) was sequentially added to a cooled (¨ 15 C)
mixture
of (R)-2-phenylglycine (3.096g, 20.48 mmol), 1N HC1 (30 mL) and methanol (40
mL). The cooling bath was removed and the reaction mixture was stirred under a
balloon of H2 for 17 hours. An additional acetaldehyde (10 mL, 178.2 mmol) was
added and stirring continued under H2 atmosphere for 24 hours [Note: the
supply of
H2 was replenished as needed throughout the reaction]. The reaction mixture
was
filtered through diatomaceous earth (Celite()), and the filtrate was
concentrated in
vacuo. The resulting crude material was recrystallized from isopropanol to
provide
the HC1 salt of (R)-2-(ethylamino)-2-phenylacetic acid as a shining white
solid
(2.846g). 1H NMR (DMSO-d6, 6=2.5 ppm, 400 MHz): 6 14.15 (br s, 1H), 9.55 (br
s,
2H), 7.55-7.48 (m, 5H), 2.88 (br m, 1H), 2.73 (br m, 1H), 1.20 (app t, J=7.2,
3H).
LC (Cond. I): RT=0.39 min; >95 % homogeneity index; LC/MS: Anal. Calcd. for
[M+H]+ C10H14NO2: 180.10; found 180.18.
A suspension of 10% Pd/C (536 mg) in methanol/H20 (3 mL/1 mL) was
added to a mixture of (R)-2-(ethylamino)-2-phenylacetic acid/HC1 (1.492g,
6.918
mmol), formaldehyde (20 mL of 37% wt. in water), 1N HC1 (20 mL) and methanol
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(23 mL). The reaction mixture was stirred under a balloon of H2 for ¨72 hours,
where the H2 supply was replenished as needed. The reaction mixture was
filtered
through diatomaceous earth (Celite ) and the filtrate was concentrated in
vacuo. The
resulting crude material was recrystallized from isopropanol (50 mL) to
provide the
HC1 salt of Cap-3 as a white solid (985 mg). 1H NMR (DMSO-d6, 6=2.5 ppm, 400
MHz): 6 10.48 (br s, 1H), 7.59-7.51 (m, 5H), 5.26 (s, 1H), 3.08 (app br s,
2H), 2.65
(br s, 3H), 1.24 (br m, 3H). LC (Cond. I): RT=0.39 min; >95 % homogeneity
index;
LC/MS: Anal. Calcd. for [M+H]+ C11H16NO2: 194.12; found 194.18; HRMS: Anal.
Calcd. for [M+H]+ C11H16NO2: 194.1180; found 194.1181.
Cap-4
so
. OH
Hy
0
C1CO2Me (3.2 mL, 41.4 mmol) was added dropwise to a cooled (ice/water)
THF (410 mL) semi-solution of (R)-tert-butyl 2-amino-2-phenylacetate/HC1
(9.877
g, 40.52 mmol) and diisopropylethylamine (14.2 mL, 81.52 mmol) over 6 min, and
stirred at similar temperature for 5.5 hours. The volatile component was
removed in
vacuo, and the residue was partitioned between water (100 mL) and ethyl
acetate
(200 mL). The organic layer was washed with 1N HC1 (25 mL) and saturated
NaHCO3 solution (30 mL), dried (MgSO4), filtered, and concentrated in vacuo.
The
resultant colorless oil was triturated from hexanes, filtered and washed with
hexanes
(100 mL) to provide (R)-tert-butyl 2-(methoxycarbonylamino)-2-phenylacetate as
a
white solid (7.7 g). 1H NMR (DMSO-d6, 6=2.5 ppm, 400 MHz): 7.98 (d, J=8.0,
1H),
7.37-7.29 (m, 5H), 5.09 (d, J=8, 1H), 3.56 (s, 3H), 1.33 (s, 9H). LC (Cond.
I):
RT=1.53 min; ¨90 % homogeneity index; LC/MS: Anal. Calcd. for [M+Na]+
C14H19NNa04: 288.12; found 288.15.
TFA (16 mL) was added dropwise to a cooled (ice/water) CH2C12 (160 mL)
solution of the above product over 7 minutes, and the cooling bath was removed
and
the reaction mixture was stirred for 20 hours. Since the deprotection was
still not
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complete, an additional TFA (1.0 mL) was added and stirring continued for an
additional 2 hours. The volatile component was removed in vacuo, and the
resulting
oil residue was treated with diethyl ether (15 mL) and hexanes (12 mL) to
provide a
precipitate. The precipitate was filtered and washed with diethyl
ether/hexanes (-1:3
ratio; 30 mL) and dried in vacuo to provide Cap-4 as a fluffy white solid
(5.57 g).
Optical rotation: -176.9 [c=3.7 mg/mL in H20; 2=589 nm]. 1H NMR (DMSO-d6,
6=2.5 ppm, 400 MHz): 6 12.84 (br s, 1H), 7.96 (d, J=8.3, 1H), 7.41-7.29 (m,
5H),
5.14 (d, J=8.3, 1H), 3.55 (s, 3H). LC (Cond. I): RT=1.01 min; >95 %
homogeneity
index; LC/MS: Anal. Calcd. for [M+H]+ CioHi2N04 210.08; found 210.17; HRMS:
Anal. Calcd. for [M+H]+ CioHi2N04 210.0766; found 210.0756.
Cap-5
0 0
. OH
N
c ____________________________________ )
A mixture of (R)- 2-phenylglycine (1.0 g, 6.62 mmol), 1,4-dibromobutane
(1.57 g, 7.27 mmol) and Na2CO3 (2.10 g, 19.8 mmol) in ethanol (40 mL) was
heated
at 100 C for 21 hours. The reaction mixture was cooled to ambient temperature
and
filtered, and the filtrate was concentrated in vacuo. The residue was
dissolved in
ethanol and acidified with 1N HC1 to pH 3-4, and the volatile component was
removed in vacuo. The resulting crude material was purified by a reverse phase
HPLC (water/methanol/TFA) to provide the TFA salt of Cap-5 as a semi-viscous
white foam (1.0 g). 1H NMR (DMSO-d6, 6=2.5, 500 MHz) 6 10.68 (br s, 1H), 7.51
(m, 5H), 5.23 (s, 1H), 3.34 (app br s, 2H), 3.05 (app br s, 2H), 1.95 (app br
s, 4H);
RT=0.30 minutes (Cond. I); >98% homogeneity index; LC/MS: Anal. Calcd. for
[M+H]+ C12H16NO2: 206.12; found 206.25.
Cap-6
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0 0
. OH
N
)
0
The TFA salt of Cap-6 was synthesized from (R)-2-phenylglycine and 1-
bromo-2-(2-bromoethoxy)ethane by using the method of preparation of Cap-5. 1H
NMR (DMSO-d6, 6=2.5, 500 MHz) 6 12.20 (br s, 1H), 7.50 (m, 5H), 4.92 (s, 1H),
3.78 (app br s, 4H), 3.08 (app br s, 2H), 2.81 (app br s, 2H); RT=0.32 minutes
(Cond.
I); >98%; LC/MS: Anal. Calcd. for [M+H]+ C12H16NO3: 222.11; found 222.20;
HRMS: Anal. Calcd. for [M+H]+ C12H16NO3: 222.1130; found 222.1121.
Cap-7
So
OH
N Cap-7a: enantiomer-1
C) Cap-7b: enantiomer-2
N
I
A CH2C12 (200 mL) solution of p-toluenesulfonyl chloride (8.65 g, 45.4
mmol) was added dropwise to a cooled (-5 C) CH2C12 (200 mL) solution of (S)-
benzyl 2-hydroxy-2-phenylacetate (10.0 g, 41.3 mmol), triethylamine (5.75 mL,
41.3
mmol) and 4-dimethylaminopyridine (0.504 g, 4.13 mmol), while maintaining the
temperature between -5 C and 0 C. The reaction was stirred at 0 C for 9
hours,
and then stored in a freezer (-25 C) for 14 hours. It was allowed to thaw to
ambient
temperature and washed with water (200 mL), 1N HC1 (100 mL) and brine (100
mL),
dried (MgSO4), filtered, and concentrated in vacuo to provide benzyl 2-pheny1-
2-
(tosyloxy)acetate as a viscous oil which solidified upon standing (16.5 g).
The chiral
integrity of the product was not checked and that product was used for the
next step
without further purification. 1H NMR (DMSO-d6, 6=2.5, 500 MHz) 6 7.78 (d, J=
8.6, 2H), 7.43-7.29 (m, 10H), 7.20 (m, 2H), 6.12 (s, 1H), 5.16 (d, J=12.5,
1H), 5.10
(d, J=12.5, 1H), 2.39 (s, 3H). RT=3.00 (Cond. III); >90% homogeneity index;
LC/MS: Anal. Calcd. for [M+H]+ C22H20Na05S: 419.09; found 419.04.
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A THF (75 mL) solution of benzyl 2-phenyl-2-(tosyloxy)acetate (6.0 g, 15.1
mmol), 1-methylpiperazine (3.36 mL, 30.3 mmol) and N,N-diisopropylethylamine
(13.2 mL, 75.8 mmol) was heated at 65 C for 7 hours. The reaction was allowed
to
cool to ambient temperature and the volatile component was removed in vacuo.
The
residue was partitioned between ethylacetate and water, and the organic layer
was
washed with water and brine, dried (MgSO4), filtered, and concentrated in
vacuo.
The resulting crude material was purified by flash chromatography (silica gel,
ethyl
acetate) to provide benzyl 2-(4-methylpiperazin-1-y1)-2-phenylacetate as an
orangish-brown viscous oil (4.56 g). Chiral HPLC analysis (Chiralcel OD-H)
indicated that the sample is a mixture of enantiomers in a 38.2 to 58.7 ratio.
The
separation of the enantiomers were effected as follow: the product was
dissolved in
120 mL of ethanol/heptane (1:1) and injected (5 mL/injection) on chiral HPLC
column (Chiracel OJ, 5 cm ID x 50 cm L, 20 p.m) eluting with 85:15
Heptane/ethanol
at 75 mL/min, and monitored at 220 nm. Enantiomer-1 (1.474 g) and enantiomer-2
(2.2149 g) were retrieved as viscous oil. 1H NMR (CDC13, 6=7.26, 500 MHz) 7.44-
7.40 (m, 2H), 7.33-7.24 (m, 6H), 7.21-7.16 (m, 2H), 5.13 (d, J=12.5, 1H), 5.08
(d,
J=12.5, 1H), 4.02 (s, 1H), 2.65-2.38 (app br s, 8H), 2.25 (s, 3H). RT=2.10
(Cond.
III); >98% homogeneity index; LC/MS: Anal. Calcd. for [M+H]+ C201-125N202:
325.19; found 325.20.
A methanol (10 mL) solution of either enantiomer of benzyl 2-(4-
methylpiperazin-1-y1)-2-phenylacetate (1.0 g, 3.1 mmol) was added to a
suspension
of 10% Pd/C (120 mg) in methanol (5.0 mL). The reaction mixture was exposed to
a
balloon of hydrogen, under a careful monitoring, for <50 minutes. Immediately
after
the completion of the reaction, the catalyst was filtered through diatomaceous
earth
(Celite ) and the filtrate was concentrated in vacuo to provide Cap-7,
contaminated
with phenylacetic acid as a tan foam (867.6 mg; mass is above the theoretical
yield).
The product was used for the next step without further purification. 1H NMR
(DMSO-d6, 6=2.5, 500 MHz) 6 7.44-7.37 (m, 2H), 7.37-7.24 (m, 3H), 3.92 (s, 11-
1),
2.63-2.48 (app. br s, 2H), 2.48-2.32 (m, 6H), 2.19 (s, 3H); RT=0.31 (Cond.
II);
>90% homogeneity index; LC/MS: Anal. Calcd. for [M+H]+ C13H19N202: 235.14;
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found 235.15; HRMS: Anal. Calcd. for [M+H]+ C13H19N202: 235.1447; found
235.1440.
The synthesis of Cap-8 and Cap-9 was conducted according to the synthesis
of Cap-7 by using appropriate amines for the SN2 displacement step (i.e., 4-
hydroxypiperidine for Cap-8 and (S)-3-fluoropyrrolidine for Cap-9) and
modified
conditions for the separation of the respective stereoisomeric intermedites,
as
described below.
Cap-8
So
OH
N
..-- ===, 8a: enantiomer-1
8b: enantiomer-2
Y
OH
The enantiomeric separation of the intermediate benzyl 2-(4-
hydroxypiperidin-1-y1)-2-phenyl acetate was effected by employing the
following
conditions: the compound (500 mg) was dissolved in ethanol/heptane (5 mL/45
mL).
The resulting solution was injected (5 mL/injection) on a chiral HPLC column
(Chiracel OJ, 2 cm ID x 25 cm L, 10 pm) eluting with 80:20 heptane/ethanol at
10
mL/min, monitored at 220 nm, to provide 186.3 mg of enantiomer-1 and 209.1 mg
of
enantiomer-2 as light-yellow viscous oils. These benzyl ester was
hydrogenolysed
according to the preparation of Cap-7 to provide Cap-8: 1H NMR (DMSO-d6,
6=2.5,
500 MHz) 7.40 (d, J=7, 2H), 7.28-7.20 (m, 3H), 3.78 (s 1H), 3.46 (m, 1H), 2.93
(m,
1H), 2.62 (m, 1H), 2.20 (m, 2H), 1.70 (m, 2H), 1.42 (m, 2H). RT=0.28 (Cond.
II);
>98% homogeneity index; LC/MS: Anal. Calcd. for [M+H]+ C13H18NO3: 236.13;
found 236.07; HRMS: Calcd. for [M+H]+ C13H18NO3: 236.1287; found 236.1283.
Cap-9
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So
OH
N 9a: diastereomer-1
\ ____________________________ Z 9b: diastereomer-2
F
The diastereomeric separation of the intermediate benzyl 2-((S)-3-
fluoropyrrolidin-1-y1)-2-phenylacetate was effected by employing the following
conditions: the ester (220 mg) was separated on a chiral HPLC column (Chiracel
OJ-
H, 0.46 cm ID x 25 cm L, 5 pm) eluting with 95% CO2/ 5% methanol with 0.1%
TFA, at 10 bar pressure, 70 mL/min flow rate, and a temperature of 35 C. The
HPLC elute for the respective stereoisomers was concentrated, and the residue
was
dissolved in CH2C12 (20 mL) and washed with an aqueous medium (10 mL water + 1
mL saturated NaHCO3 solution). The organic phase was dried (MgSO4), filtered,
and concentrated in vacuo to provide 92.5 mg of fraction-1 and 59.6 mg of
fraction-2.
These benzyl esters were hydrogenolysed according to the preparation of Cap-7
to
prepare Caps 9a and 9b. Cap-9a (diastereomer-1; the sample is a TFA salt as a
result
of purification on a reverse phase HPLC using H20/methanol/TFA solvent): 1H
NMR
(DMSO-d6, 6=2.5, 400 MHz) 7.55-7.48 (m, 5H), 5.38 (d of m, J=53.7, 1H), 5.09
(br
s, 1H), 3.84-2.82 (br m, 4H), 2.31-2.09 (m, 2H). RT=0.42 (Cond. I); >95%
homogeneity index; LC/MS: Anal. Calcd. for [M+H]+ C12H15FN02: 224.11; found
224.14; Cap-9b (diastereomer-2): 1H NMR (DMSO-d6, 6=2.5, 400 MHz) 7.43-7.21
(m, 5H), 5.19 (d of m, J=55.9, 1H), 3.97 (s, 1H), 2.95-2.43 (m, 4H), 2.19-1.78
(m,
2H). RT=0.44 (Cond. I); LC/MS: Anal. Calcd. for [M+H]+ C12H15FN02: 224.11;
found 224.14.
Cap-10
0 ,
1
HO)N
0
To a solution of D-proline (2.0 g, 17 mmol) and formaldehyde (2.0 mL of
37% wt. in H20) in methanol (15 mL) was added a suspension of 10% Pd/C (500
mg) in methanol (5 mL). The mixture was stirred under a balloon of hydrogen
for 23
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hours. The reaction mixture was filtered through diatomaceous earth (Celite )
and
concentrated in vacuo to provide Cap-10 as an off-white solid (2.15 g). 1H NMR
(DMSO-d6, 6=2.5, 500 MHz) 3.42 (m, 1H), 3.37 (dd, J=9.4, 6.1, 1H), 2.85-2.78
(m,
1H), 2.66 (s, 3H), 2.21-2.13 (m, 1H), 1.93-1.84 (m, 2H), 1.75-1.66 (m, 1H).
RT=0.28 (Cond. II); >98% homogeneity index; LC/MS: Anal. Calcd. for [M+H]+
C6H12NO2: 130.09; found 129.96.
Cap-11
0 1
HO)1.31
F
A mixture of (2S,4R)-4-fluoropyrrolidine-2-carboxylic acid (0.50 g, 3.8
mmol), formaldehyde (0.5 mL of 37% wt. in H20), 12 N HC1 (0.25 mL) and 10%
Pd/C (50 mg) in methanol (20 mL) was stirred under a balloon of hydrogen for
19
hours. The reaction mixture was filtered through diatomaceous earth (Celite )
and
the filtrate was concentrated in vacuo. The residue was recrystallized from
isopropanol to provide the HC1 salt of Cap-11 as a white solid (337.7 mg). 1H
NMR
(DMSO-d6, 6=2.5, 500 MHz) 5.39 (d m, J=53.7, 1H), 4.30 (m, 1H), 3.90 (ddd,
J=31.5, 13.5, 4.5, 1H), 3.33 (dd, J=25.6, 13.4, 1H), 2.85 (s, 3H), 2.60-2.51
(m, 1H),
2.39-2.26 (m, 1H). RT=0.28 (Cond. II); >98% homogeneity index; LC/MS: Anal.
Calcd. for [M+H]+ C6H11FN02: 148.08; found 148.06.
Cap-12 (same as cap 52)
0
H
N J.L
0
y , OH
0 =
L-Alanine (2.0 g, 22.5 mmol) was dissolved in 10% aqueous sodium
carbonate solution (50 mL), and a THF (50 mL) solution of methyl chloroformate
(4.0 mL) was added to it. The reaction mixture was stirred under ambient
conditions
for 4.5 hours and concentrated in vacuo. The resulting white solid was
dissolved in
water and acidified with 1N HC1 to a pH ¨ 2-3. The resulting solutions was
extracted
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with ethyl acetate (3 x 100 mL), and the combined organic phase was dried
(Na2SO4), filtered, and concentrated in vacuo to provide a colorless oil (2.58
g). 500
mg of this material was purified by a reverse phase HPLC (H20/methanol/TFA) to
provide 150 mg of Cap-12 as a colorless oil. 1H NMR (DMSO-d6, 6=2.5, 500 MHz)
7.44 (d, J=7.3, 0.8H), 7.10 (br s, 0.2H), 3.97 (m, 1H), 3.53 (s, 3H), 1.25 (d,
J=7.3,
3H).
Cap-13
0
N H
A mixture of L-alanine (2.5 g, 28 mmol), formaldehyde (8.4 g, 37 wt. %), 1N
HC1 (30 mL) and 10% Pd/C (500 mg) in methanol (30 mL) was stirred under a
hydrogen atmosphere (50 psi) for 5 hours. The reaction mixture was filtered
through
diatomaceous earth (Celite ) and the filtrate was concentrated in vacuo to
provide the
HC1 salt of Cap-13 as an oil which solidified upon standing under vacuum (4.4
g; the
mass is above theoretical yield). The product was used without further
purification.
1H NMR (DMSO-d6, 6=2.5, 500 MHz) 6 12.1 (br s, 1H), 4.06 (q, J=7.4, 1H), 2.76
(s,
6H), 1.46 (d, J=7.3, 3H).
Cap-14
NH2
N
CO 1 22t-Bu , las
CO2t-Bu ¨1" CO2H
Cap-14
Step 1: A mixture of (R)-(-)-D-phenylglycine tert-butyl ester (3.00 g, 12.3
mmol), NaBH3CN (0.773 g, 12.3 mmol), KOH (0.690 g, 12.3 mmol) and acetic acid
(0.352 mL, 6.15 mmol) were stirred in methanol at 0 C. To this mixture was
added
glutaric dialdehyde (2.23 mL, 12.3 mmol) dropwise over 5 minutes. The reaction
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removed and the residue was partitioned with 10% aqueous NaOH and ethyl
acetate.
The organic phase was separated, dried (MgSO4), filtered and concentrated to
dryness to provide a clear oil. This material was purified by reverse-phase
preparative HPLC (Primesphere C-18, 30 x 100mm; CH3CN-H20-0.1% TFA) to give
the intermediate ester (2.70 g, 56%) as a clear oil. 1H NMR (400 MHz, CDC13) 6
7.53-7.44 (m, 3H), 7.40-7.37 (m, 2H), 3.87 (d, J=10.9 Hz, 1H), 3.59 (d, J=10.9
Hz,
1H), 2.99 (t, J=11.2 Hz, 1H), 2.59 (t, J=11.4 Hz, 1H), 2.07-2.02 (m, 2H), 1.82
(d,
J=1.82 Hz, 3H), 1.40 (s, 9H). LC/MS: Anal. Calcd. for C12H25NO2: 275; found:
276 (M+H)+.
Step 2: To a stirred solution of the intermediate ester (1.12g, 2.88mmol) in
dichloromethane (10 mL) was added TFA (3 mL). The reaction mixture was stirred
at ambient temperature for 4 hours and then it was concentrated to dryness to
give a
light yellow oil. The oil was purified using reverse-phase preparative HPLC
(Primesphere C-18, 30 x 100mm; CH3CN-H20-0.1% TFA). The appropriate
fractions were combined and concentrated to dryness in vacuo. The residue was
then
dissolved in a minimum amount of methanol and applied to applied to MCX LP
extraction cartridges (2 x 6 g). The cartridges were rinsed with methanol (40
mL)
and then the desired compound was eluted using 2M ammonia in methanol (50 mL).
Product-containing fractions were combined and concentrated and the residue
was
taken up in water. Lyophilization of this solution provided the title compound
(0.492
g, 78%) as a light yellow solid. 1H NMR (DMSO-d6) 6 7.50 (s, 5H), 5.13 (s,
1H),
3.09 (br s, 2H), 2.92-2.89 (m, 2H), 1.74 (m, 4H), 1.48 (br s, 2H). LC/MS:
Anal.
Calcd. for C13H12NO2: 219; found: 220 (M+H)+.
Cap-15
0 0
OH 1 401 0 Me 2 0 0 Me
Br Br 0 SI
---11%.
..-- -..
A BHO Me C
HO Me
(R)-Cap-15
Step 1: (S)-1-Phenylethyl 2-bromo-2-phenylacetate: To a mixture of a-
bromophenylacetic acid (10.75 g, 0.050 mol), (S)-(-)-1-phenylethanol (7.94 g,
0.065
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mol) and DMAP (0.61 g, 5.0 mmol) in dry dichloromethane (100 mL) was added
solid EDCI (12.46 g, 0.065 mol) all at once. The resulting solution was
stirred at
room temperature under Ar for 18 hours and then it was diluted with ethyl
acetate,
washed (H20 x 2, brine), dried (Na2SO4), filtered, and concentrated to give a
pale
yellow oil. Flash chromatography (Si02/ hexane-ethyl acetate, 4:1) of this oil
provided the title compound (11.64 g, 73%) as a white solid. 1H NMR (400 MHz,
CDC13) 6 7.53-7.17 (m, 10H), 5.95 (q, J=6.6 Hz, 0.5H), 5.94 (q, J=6.6 Hz,
0.5H),
5.41 (s, 0.5H), 5.39 (s, 0.5H), 1.58 (d, J=6.6 Hz, 1.5H), 1.51 (d, J=6.6 Hz,
1.5H).
Step 2: (S)-1-Phenylethyl (R)-2-(4-hydroxy-4-methylpiperidin-1-y1)- 2-
phenylacetate: To a solution of (S)-1-phenylethyl 2-bromo-2-phenylacetate
(0.464 g,
1.45 mmol) in THF (8 mL) was added triethylamine (0.61 mL, 4.35 mmol),
followed
by tetrabutylammonium iodide (0.215 g, 0.58 mmol). The reaction mixture was
stirred at room temperature for 5 minutes and then a solution of 4-methy1-4-
hydroxypiperidine (0.251 g, 2.18 mmol) in THF (2 mL) was added. The mixture
was
stirred for 1 hour at room temperature and then it was heated at 55-60 C (oil
bath
temperature) for 4 hours. The cooled reaction mixture was then diluted with
ethyl
acetate (30 mL), washed (H20 x2, brine), dried (MgSO4), filtered and
concentrated.
The residue was purified by silica gel chromatography (0-60% ethyl acetate-
hexane)
to provide first the (S,R)-isomer of the title compound (0.306 g, 60%) as a
white
solid and then the corresponding (S,S)-isomer (0.120 g, 23%), also as a white
solid.
(S,R)-isomer: 1H NMR (CD30D) 6 7.51-7.45 (m, 2H), 7.41-7.25 (m, 8H), 5.85 (q,
J=6.6 Hz, 1H), 4.05 (s, 1H), 2.56-2.45 (m, 2H), 2.41-2.29 (m, 2H), 1.71-1.49
(m,
4H), 1.38 (d, J=6.6 Hz, 3H), 1.18 (s, 3H). LCMS: Anal. Calcd. for C22H22NO3:
353; found: 354 (M+H)+. (S,S)-isomer: 1H NMR (CD30D) 6 7.41-7.30 (m, 5H),
7.20-7.14 (m, 3H), 7.06-7.00 (m, 2H), 5.85 (q, J=6.6 Hz, 1H), 4.06 (s, 1H),
2.70-2.60
(m, 1H), 2.51 (dt, J=6.6, 3.3 Hz, 1H), 2.44-2.31 (m, 2H), 1.75-1.65 (m, 1H),
1.65-
1.54 (m, 3H), 1.50 (d, J=6.8 Hz, 3H), 1.20 (s, 3H). LCMS: Anal. Calcd. for
C22H22NO3: 353; found: 354 (M+H)+.
Step 3: (R)-2-(4-Hydroxy-4-methylpiperidin-1-y1)-2-phenylacetic acid: To a
solution of (S)-1-phenylethyl (R)-2-(4-hydroxy-4-methylpiperidin-1-y1)-2-
phenylacetate (0.185 g, 0.52 mmol) in dichloromethane (3 mL) was added
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trifluoroacetic acid (1 mL) and the mixture was stirred at room temperature
for 2
hours. The volatiles were subsequently removed in vacuo and the residue was
purified by reverse-phase preparative HPLC (Primesphere C-18, 20 x 100mm;
CH3CN-H20-0.1% TFA) to give the title compound (as TFA salt) as a pale bluish
solid (0,128 g, 98%). LCMS: Anal. Calcd. for C14H19NO3: 249; found: 250
(M+H)+.
Cap-16
40 0 1 0
OH 0 Me 40 0
. OH
,
F FN FN
N
--- ...
Fl
A \/ B \/
(R)-Cap-1 6
Step 1: (S)-1-Phenylethyl 2-(2-fluorophenyl)acetate: A mixture of 2-
fluorophenylacetic acid (5.45 g, 35.4 mmol), (S)-1-phenylethanol (5.62 g, 46.0
mmol), EDCI (8.82 g, 46.0 mmol) and DMAP (0.561 g, 4.60 mmol) in CH2C12 (100
mL) was stirred at room temperature for 12 hours. The solvent was then
concentrated and the residue partitioned with H20-ethyl acetate. The phases
were
separated and the aqueous layer back-extracted with ethyl acetate (2x). The
combined organic phases were washed (H20, brine), dried (Na2SO4), filtered,
and
concentrated in vacuo. The residue was purified by silica gel chromatography
(Biotage/ 0-20% ethyl acetate-hexane) to provide the title compound as a
colorless
oil (8.38 g, 92%). 1H NMR (400 MHz, CD30D) 6 7.32-7.23 (m, 7H), 7.10-7.04 (m,
2), 5.85 (q, J=6.5 Hz, 1H), 3.71 (s, 2H), 1.48 (d, J=6.5 Hz, 3H).
Step 2: (R)-((S)-1-Phenylethyl) 2-(2-fluoropheny1)-2-(piperidin-1-y1)acetate:
To a solution of (S)-1-phenylethyl 2-(2-fluorophenyl)acetate (5.00 g, 19.4
mmol) in
THF (1200 mL) at 0 C was added DBU (6.19 g, 40.7 mmol) and the solution was
allowed to warm to room temperature while stirring for 30 minutes. The
solution
was then cooled to -78 C and a solution of CBr4(13.5 g, 40.7 mmol) in THF
(100
mL) was added and the mixture was allowed to warm to -10 C and stirred at
this
temperature for 2 hours. The reaction mixture was quenched with saturated aq.
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NH4C1 and the layers were separated. The aqueous layer was back-extracted with
ethyl acetate (2x) and the combined organic phases were washed (H20, brine),
dried
(Na2SO4), filtered, and concentrated in vacuo. To the residue was added
piperidine
(5.73 mL, 58.1 mmol) and the solution was stirred at room temperature for 24
hours.
The volatiles were then concentrated in vacuo and the residue was purified by
silica
gel chromatography (Biotage/ 0-30% diethyl ether-hexane) to provide a pure
mixture
of diastereomers (2:1 ratio by 1H NMR) as a yellow oil (2.07 g, 31%), along
with
unreacted starting material (2.53 g, 51%). Further chromatography of the
diastereomeric mixture (Biotage/ 0-10% diethyl ether-toluene) provided the
title
compound as a colorless oil (0.737 g, 11%). 1H NMR (400 MHz, CD30D) 6 7.52
(ddd, J=9.4, 7.6, 1.8 Hz, 1H), 7.33 ¨ 7.40 (m, 1), 7.23 ¨ 7.23 (m, 4H), 7.02 ¨
7.23 (m,
4H), 5.86 (q, J=6.6 Hz, 1H), 4.45 (s, 1H), 2.39 ¨2.45 (m, 4H), 1.52 ¨ 1.58 (m,
4H),
1.40 ¨ 1.42 (m, 1H), 1.38 (d, J=6.6 Hz, 3H). LCMS: Anal. Calcd. for C211-
124FN02:
341; found: 342 (M+H)+.
Step 3: (R)-2-(2-fluoropheny1)-2-(piperidin-1-y1)acetic acid: A mixture of
(R)-((S)-1-phenylethyl) 2-(2-fluoropheny1)-2-(piperidin-1-y1)acetate (0.737 g,
2.16
mmol) and 20% Pd(OH)2/C (0.070 g) in ethanol (30 mL) was hydrogenated at room
temperature and atmospheric pressure (H2 balloon) for 2 hours. The solution
was
then purged with Ar, filtered through diatomaceous earth (Celite()), and
concentrated
in vacuo. This provided the title compound as a colorless solid (0.503 g,
98%). 1H
NMR (400 MHz, CD30D) 6 7.65 (ddd, J=9.1, 7.6, 1.5 Hz, 1H), 7.47-7.53 (m, 1H),
7.21-7.30 (m, 2H), 3.07-3.13 (m, 4H), 1.84 (br s, 4H), 1.62 (br s, 2H). LCMS:
Anal.
Calcd. for C13H16FN02: 237; found: 238 (M+H)+.
Cap-17
40 0 Me 1 40 0 Me
2 0 0
OH
N
Br
N
..-- -...
AB
HO Ph HO Ph
(R)-Cap-17
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Step 1: (S)-1-Phenylethyl (R)-2-(4-hydroxy-4-phenylpiperidin-l-y1)- 2-
phenylacetate: To a solution of (S)-1-phenylethyl 2-bromo-2-phenylacetate
(1.50 g,
4.70 mmol) in THF (25 mL) was added triethylamine (1.31 mL, 9.42 mmol),
followed by tetrabutylammonium iodide (0.347 g, 0.94 mmol). The reaction
mixture
was stirred at room temperature for 5 minutes and then a solution of 4-pheny1-
4-
hydroxypiperidine (1.00 g, 5.64 mmol) in THF (5 mL) was added. The mixture was
stirred for 16 hours and then it was diluted with ethyl acetate (100 mL),
washed (H2O
x2, brine), dried (MgSO4), filtered and concentrated. The residue was purified
on a
silica gel column (0-60% ethyl acetate-hexane) to provide an approximately 2:1
mixture of diastereomers, as judged by 1H NMR. Separation of these isomers was
performed using supercritical fluid chromatography (Chiralcel OJ-H, 30 x
250mm;
20% ethanol in CO2 at 35 C), to give first the (R)-isomer of the title
compound
(0.534 g, 27%) as a yellow oil and then the corresponding (S)-isomer (0.271 g,
14%),
also as a yellow oil. (S,R)-isomer: 1H NMR (400 MHz, CD30D) 6 7.55-7.47 (m,
4H), 7.44-7.25 (m, 10H), 7.25-7.17 (m, 1H), 5.88 (q, J=6.6 Hz, 1H), 4.12 (s,
1H),
2.82-2.72 (m, 1H), 2.64 (dt, J=11.1, 2.5 Hz, 1H), 2.58-2.52 (m, 1H), 2.40 (dt,
J=11.1, 2.5 Hz, 1H), 2.20 (dt, J=12.1, 4.6 Hz, 1H), 2.10 (dt, J=12.1, 4.6 Hz,
1H),
1.72-1.57 (m, 2H), 1.53 (d, J=6.5 Hz, 3H). LCMS: Anal. Calcd. for C271-129NO3:
415; found: 416 (M+H)+; (S,S)-isomer: H1NMR (400 MHz, CD30D) 6 7.55-7.48
(m, 2H), 7.45-7.39 (m, 2H), 7.38-7.30 (m, 5H), 7.25-7.13 (m, 4H), 7.08-7.00
(m,
2H), 5.88 (q, J=6.6 Hz, 1H), 4.12 (s, 1H), 2.95-2.85 (m, 1H), 2.68 (dt,
J=11.1, 2.5
Hz, 1H), 2.57-2.52 (m, 1H), 2.42 (dt, J=11.1, 2.5 Hz, 1H), 2.25 (dt, J=12.1,
4.6 Hz,
1H), 2.12 (dt, J=12.1, 4.6 Hz, 1H), 1.73 (dd, J=13.6, 3.0 Hz, 1H), 1.64 (dd,
J=13.6,
3.0 Hz, 1H), 1.40 (d, J=6.6 Hz, 3H). LCMS: Anal. Calcd. for C27H29NO3: 415;
found: 416 (M+H)+.
The following esters were prepared in similar fashion:
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Intermediate-17a Diastereomer 1: 1H NMR
0y0 el (500 MHz, DMSO-d6) 6
N ppm 1.36 (d, J=6.41 Hz,
C )
N 3H) 2.23-2.51 (m, 4H)
lel0 3.35 (s, 4H) 4.25 (s, 1H)
0 0 5.05 (s, 2H) 5.82 (d,
J=6.71 Hz, 1H) 7.15-7.52
(m, 15H).
LCMS: Anal. Calcd. for:
C28H30N204 458.22;
Found: 459.44 (M+H)+.
Diastereomer 2: 1H NMR
(500 MHz, DMSO-d6) 6
ppm 1.45 (d, J=6.71 Hz,
3H) 2.27-2.44 (m, 4H)
3.39 (s, 4H) 4.23 (s, 1H)
5.06 (s, 2H) 5.83 (d,
J=6.71 Hz, 1H) 7.12 (dd,
J=6.41, 3.05 Hz, 2H)
7.19-7.27 (m, 3H) 7.27-
7.44 (m, 10H).
LCMS: Anal. Calcd. for:
C28H30N204 458.22;
Found: 459.44 (M+H)+.
Intermediate -17b H Diasteromer 1: RT=11.76
N 0
Cminutes (Cond'n II);
N
LCMS: Anal. Calcd. for:
0
01 * C20I-122N203 338.16
0
el Found: 339.39 (M+H)+;
Diastereomer 2:
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RT=10.05 minutes
(Cond'n II); LCMS: Anal.
Calcd. for: C20H22N203
338.16; Found: 339.39
(M+H)+.
Intermediate -17c I Diastereomer 1: TR=4.55
N
C ) minutes (Cond'n I);
N LCMS: Anal. Calcd. for:
lel0
*
C21H26N202 338.20
o
lei Found: 339.45 (M+H)+;
Diastereomer 2: TR=6.00
minutes (Cond'n I);
LCMS: Anal. Calcd. for:
C21H26N202 338.20
Found: 339.45 (M+H)+.
Intermediate -17d
40 Diastereomer 1: RT=7.19
minutes (Cond'n I);
LCMS: Anal. Calcd. for:
N C27H29NO2 399.22
110I * 0 Found: 400.48 (M+H)+;
0
I. Diastereomer 2: RT=9.76
minutes (Cond'n I);
LCMS: Anal. Calcd. for:
C27H29NO2 399.22
Found: 400.48 (M+H)+.
Chiral SFC Conditions for determining retention time
Condition I
Column: Chiralpak AD-H Column, 4.62x50 mm, 5pm
Solvents: 90% CO2-10% methanol with 0.1%DEA
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Temp: 35 C
Pressure: 150 bar
Flow rate: 2.0 mL/min.
UV monitored @ 220 nm
Injection: 1.0 mg/3mL methanol
Condition II
Column: Chiralcel OD-H Column, 4.62x50 mm, 51.tm
Solvents: 90% CO2-10% methanol with 0.1%DEA
Temp: 35 C
Pressure: 150 bar
Flow rate: 2.0 mL/min.
UV monitored @ 220 nm
Injection: 1.0 mg/mL methanol
Cap 17, Step 2; (R)-2-(4-Hydroxy-4-phenylpiperidin-1-y1)-2-phenylacetic
acid: To a solution of (S)-1-phenylethyl (R)-2-(4-hydroxy-4-phenylpiperidin-1-
y1)-
2-phenylacetate (0.350 g, 0.84 mmol) in dichloromethane (5 mL) was added
trifluoroacetic acid (1 mL) and the mixture was stirred at room temperature
for 2
hours. The volatiles were subsequently removed in vacuo and the residue was
purified by reverse-phase preparative HPLC (Primesphere C-18, 20 x 100mm;
CH3CN-H20-0.1% TFA) to give the title compound (as TFA salt) as a white solid
(0.230 g, 88%). LCMS: Anal. Calcd. for Ci9H2iNO3: 311.15; found: 312 (M+H)+.
The following carboxylic acids were prepared in optically pure form in a
similar
fashion:
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Cap-17a RT=2.21 (Cond'n II);
OyO el 1H NMR (500 MHz,
N DMSO-d6) 6 ppm 2.20-
( )
N 2.35 (m, 2H) 2.34-2.47
0 * OH (m, 2H) 3.37 (s, 4H)
0
3.71 (s, 1H) 5.06 (s,
2H) 7.06-7.53 (m,
10H). LCMS: Anal.
Calcd. for: C20H22N204
354.16; Found: 355.38
(M+H)+.
Cap-17b H RT=0.27 (Cond'n III);
N 0
CN: LCMS: Anal. Calcd.
for: C12H14N203
0 * 0 OH
234.10; Found: 235.22
(M+H)+.
Cap-17c I RT=0.48 (Cond'n II);
N
C ) LCMS: Anal. Calcd.
N for: C13H181\1202
0 * 0 OH
234.14; Found: 235.31
(M+H)+.
Cap-17d
01 RT=2.21 (Cond'n I);
LCMS: Anal. Calcd.
for: C19H21NO2 295.16;
N Found: 296.33 (M+H)+.
0 * OH
0
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LCMS Conditions for determining retention time
Condition I
Column: Phenomenex-Luna 4.6 X 50 mm S10
Start % B=0
Fianl % B=100
Gradient Time=4 min
Flow Rate=4 mL/min
Wavelength=220
Solvent A=10% methanol ¨ 90% H20 ¨ 0.1% TFA
Solvent B=90% methanol ¨ 10% H20 ¨ 0.1% TFA
Condition II
Column: Waters-Sunfire 4.6 X 50 mm S5
Start % B=0
Fianl % B=100
Gradient Time=2 min
Flow Rate=4 mL/min
Wavelength=220
Solvent A=10% methanol ¨ 90% H20 ¨ 0.1% TFA
Solvent B=90% methanol ¨ 10% H20 ¨ 0.1% TFA
Condition III
Column: Phenomenex 10 . 3.0 X 50 mm
Start % B=0
Fianl % B=100
Gradient Time=2 min
Flow Rate=4 mL/min
Wavelength=220
Solvent A=10% methanol ¨ 90% H20 ¨ 0.1% TFA
Solvent B=90% methanol ¨ 10% H20 ¨ 0.1% TFA
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Cap-18
X N N
rrOEt 2
________________________________ rrOEt 3
rrOH
N 0
N 0 N 0
A: X = H j C cap-18
1
B: X = Br
Step 1; (R,S)-Ethyl 2-(4-pyridy1)-2-bromoacetate: To a solution of ethyl 4-
pyridylacetate (1.00 g, 6.05 mmol) in dry THF (150 mL) at 0 C under argon was
added DBU (0.99 mL, 6.66 mmol). The reaction mixture was allowed to warm to
room temperature over 30 minutes and then it was cooled to -78 C. To this
mixture
was added CBr4 (2.21 g, 6.66 mmol) and stirring was continued at -78 C for 2
hours.
The reaction mixture was then quenched with sat. aq. NH4C1 and the phases were
separated. The organic phase was washed (brine), dried (Na2SO4), filtered, and
concentrated in vacuo. The resulting yellow oil was immediately purified by
flash
chromatography (Si02/ hexane-ethyl acetate, 1:1) to provide the title compound
(1.40
g, 95%) as a somewhat unstable yellow oil. 1H NMR (400 MHz, CDC13) 6 8.62 (dd,
J=4.6, 1.8 Hz, 2H), 7.45 (dd, J=4.6, 1.8 Hz, 2H), 5.24 (s, 1H), 4.21-4.29 (m,
2H),
1.28 (t, J=7.1 Hz, 3H). LCMS: Anal. Calcd. for C9H10BrNO2: 242, 244; found:
243, 245 (M+H)+.
Step 2; (R,S)-Ethyl 2-(4-pyridy1)-2-(N,N-dimethylamino)acetate: To a
solution of (R,S)-ethyl 2-(4-pyridy1)-2-bromoacetate (1.40 g, 8.48 mmol) in
DMF (10
mL) at room temperature was added dimethylamine (2M in THF, 8.5 mL, 17.0
mmol). After completion of the reaction (as judged by thin layer
chromatography)
the volatiles were removed in vacuo and the residue was purified by flash
chromatography (Biotage, 40+M Si02 column; 50%-100% ethyl acetate-hexane) to
provide the title compound (0.539 g, 31%) as a light yellow oil. 1H NMR (400
MHz,
CDC13) 6 8.58 (d, J=6.0 Hz, 2H), 7.36 (d, J=6.0 Hz, 2H), 4.17 (m, 2H), 3.92
(s, 1H),
2.27 (s, 6H), 1.22 (t, J=7.0 Hz). LCMS: Anal. Calcd. for C11H16N202: 208;
found:
209 (M+H)+.
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Step 3; (R,S)-2-(4-Pyridy1)-2-(N,N-dimethylamino)acetic acid: To a solution
of (R,S)-ethyl 2-(4-pyridy1)-2-(N,N-dimethylamino)acetate (0.200 g, 0.960
mmol) in
a mixture of THF-methanol-H20 (1:1:1, 6 mL) was added powdered LiOH (0.120 g,
4.99 mmol) at room temperature. The solution was stirred for 3 hours and then
it was
acidified to pH 6 using 1N HC1. The aqueous phase was washed with ethyl
acetate
and then it was lyophilized to give the dihydrochloride of the title compound
as a
yellow solid (containing LiC1). The product was used as such in subsequent
steps.
1H NMR (400 MHz, DMSO-d6) 6 8.49 (d, J=5.7 Hz, 2H), 7.34 (d, J=5.7 Hz, 2H),
3.56 (s, 1H), 2.21 (s, 6H).
The following examples were prepared in similar fashion using the method
described above;
NMe2 LCMS: Anal. Calcd. for
Cap-19 CO2H
I C9H12N202: 180; found: 181
N (M+H)+.
LCMS: no ionization. 1H NMR
(400 MHz, CD30D) 6 8.55 (d,
NMe2
J=4.3 Hz, 1H), 7.84 (app t,
Cap-20
CO2H
I J=5.3 Hz, 1H), 7.61 (d, J=7.8
N
Hz, 1H), 7.37 (app t, J=5.3 Hz,
1H), 4.35 (s, 1H), 2.60 (s, 6H).
NMe2 LCMS: Anal. Calcd. for
Cap-21 1 CO2H C9H11C1N202: 214, 216; found:
CI N 215, 217 (M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-22 0 C 02 H C10H12N204: 224; found: 225
02N (M+H)+.
Nme2
LCMS: Anal. Calcd. for
0 CO2H
Cap-23 C14H15NO2: 229; found: 230
lei (M+H)+.
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NMe2 LCMS: Anal. Calcd. for
C 0 Cap-24 F3 CO2H C11F112F3NO2: 247; found: 248
(M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-25 0 CO2H C11H12F3NO2: 247; found: 248
CF3 (M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-26 110 CO2H C10H12FN02: 197; found: 198
F (M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-27 F 40
CO2H C10H12FN02: 247; found: 248
(M+H)+.
NMe2 LCMS: Anal. Calcd. for
I. Cap-28 CI CO2H C10H12C1NO2: 213; found: 214
(M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-29 40 CO2H C10th2C1NO2: 213; found: 214
CI (M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-30 40 CO2H C10H12C1NO2: 213; found: 214
CI (M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-31 s002H C8H12N202S: 200; found: 201
--7.---N
(M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-32
C.7)'CO2H C8FI11NO2S: 185; found: 186
\ S (M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-33s3)CO2H FIH C8NO2S: 185; found: 186
-
(M+H)+.
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NMe2 LCMS: Anal. Calcd. for
Cap-34 CO2H C11H12N203: 220; found: 221
(M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-35 CO2H C12H13NO2S: 235; found: 236
(M+H)+.
NMe2 LCMS: Anal. Calcd. for
Cap-36 = CO2H C12H14N202S: 250; found: 251
(M+H)+.
Cap-37
N N
M .r0Et 1 2
e2N
OEt OH
0 Me2N Me2N
0 = HCI 0
A
cap-37
Step 1; (R, S)-Ethyl 2-(quinolin-3-y1)-2-(N,N-dimethylamino)-acetate: A
mixture of ethyl N,N-dimethylaminoacetate (0.462 g, 3.54 mmol), K3PO4 (1.90 g,
8.95 mmol), Pd(t-Bu3P)2 (0.090 g, 0.176 mmol) and toluene (10 mL) was degassed
with a stream of Ar bubbles for 15 minutes. The reaction mixture was then
heated at
100 C for 12 hours, after which it was cooled to room temperature and poured
into
H20. The mixture was extracted with ethyl acetate (2x) and the combined
organic
phases were washed (H20, brine), dried (Na2SO4), filtered, and concentrated in
vacuo. The residue was purified first by reverse-phase preparative HPLC
(Primesphere C-18, 30 x 100mm; CH3CN-H20-5 mM NH40Ac) and then by flash
chromatography (Si02/ hexane-ethyl acetate, 1:1) to provide the title compound
(0.128 g, 17%) as an orange oil. 1FINMR (400 MHz, CDC13) 6 8.90 (d, J=2.0 Hz,
1H), 8.32 (d, J=2.0 Hz, 1H), 8.03-8.01 (m, 2H), 7.77 (ddd, J=8.3, 6.8, 1.5 Hz,
1H),
7.62 (ddd, J=8.3, 6.8, 1.5 Hz, 1H), 4.35 (s, 1H), 4.13 (m, 2H), 2.22 (s, 6H),
1.15 (t,
J=7.0 Hz, 3H). LCMS: Anal. Calcd. for C15H18N202: 258; found: 259 (M+H)+.
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Step 2; (R,S) 2-(Quinolin-3-y1)-2-(N,N-dimethylamino)acetic acid: A
mixture of (R,S)-ethyl 2-(quinolin-3-y1)-2-(N,N-dimethylamino)acetate (0.122
g,
0.472 mmol) and 6M HC1 (3 mL) was heated at 100 C for 12 hours. The solvent
was removed in vacuo to provide the dihydrochloride of the title compound
(0.169 g,
>100%) as a light yellow foam. The unpurified material was used in subsequent
steps without further purification. LCMS: Anal. Calcd. for C13H14N202: 230;
found: 231 (M+H)+.
Cap-38
101 0 IS
OH 1 N 0 Me 0
0 IS 0 F
2 F N
OH
--- --,
¨1-
+
F NR1R2 10 0 Me
0 0
, 0 0
A F . OH
Fl
F F1-
.--. .....
B Cap-38
Step 1; (R)-((S)-1-phenylethyl) 2-(dimethylamino)-2-(2-fluorophenyl)acetate
and (S)-((S)-1-phenylethyl) 2-(dimethylamino)-2-(2-fluorophenyl)acetate: To a
mixture of (RS)-2-(dimethylamino)-2-(2-fluorophenyl)acetic acid (2.60 g, 13.19
mmol), DMAP (0.209 g, 1.71 mmol) and (S)-1-phenylethanol (2.09 g, 17.15 mmol)
in CH2C12 (40 mL) was added EDCI (3.29 g, 17.15 mmol) and the mixture was
allowed to stir at room temperature for 12 hours. The solvent was then removed
in
vacuo and the residue partitioned with ethyl acetate-H20. The layers were
separated,
the aqueous layer was back-extracted with ethyl acetate (2x) and the combined
organic phases were washed (H20, brine), dried (Na2SO4), filtered, and
concentrated
in vacuo. The residue was purified by silica gel chromatography (Biotage/ 0-
50%
diethyl ether-hexane). The resulting pure diastereomeric mixture was then
separated
by reverse-phase preparative HPLC (Primesphere C-18, 30 x 100mm; CH3CN-H20-
0.1% TFA) to give first (S)-1-phenethyl (R)-2-(dimethylamino)-2-(2-
fluorophenyl)acetate (0.501 g, 13%) and then (S)-1-phenethyl (S)-2-
(dimethylamino)-2-(2-fluoropheny1)-acetate (0.727 g. 18%), both as their TFA
salts.
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(S,R)-isomer: 1H NMR (400 MHz, CD30D) 6 7.65-7.70 (m, 1H), 7.55-7.60 (ddd,
J=9.4, 8.1, 1.5 Hz, 1H), 7.36-7.41 (m, 2H), 7.28-7.34 (m, 5H), 6.04 (q, J=6.5
Hz,
1H), 5.60 (s, 1H), 2.84 (s, 6H), 1.43 (d, J=6.5 Hz, 3H). LCMS: Anal. Calcd.
for
C18H20FN02: 301; found: 302 (M+H)+; (S,S)-isomer: 1H NMR (400 MHz, CD30D) 6
7.58-7.63 (m, 1H), 7.18-7.31 (m, 6H), 7.00 (dd, J=8.5, 1.5 Hz, 2H), 6.02 (q,
J=6.5
Hz, 1H), 5.60 (s, 1H), 2.88 (s, 6H), 1.54 (d, J=6.5 Hz, 3H). LCMS: Anal.
Calcd.
for C18H20FN02: 301; found: 302 (M+H)+.
Step 2; (R)-2-(dimethylamino)-2-(2-fluorophenyl)acetic acid: A mixture of
(R)-((S)-1-phenylethyl) 2-(dimethylamino)-2-(2-fluorophenyl)acetate TFA salt
(1.25
g, 3.01 mmol) and 20% Pd(OH)2/C (0.125 g) in ethanol (30 mL) was hydrogenated
at
room temperature and atmospheric pressure (H2 balloon) for 4 hours. The
solution
was then purged with Ar, filtered through diatomaceous earth (Celite()), and
concentrated in vacuo. This gave the title compound as a colorless solid
(0.503 g,
98%). 1H NMR (400 MHz, CD30D) 6 7.53-7.63 (m, 2H), 7.33-7.38 (m, 2H), 5.36
(s, 1H), 2.86 (s, 6H). LCMS: Anal. Calcd. for C10H12FN02: 197; found: 198
(M+H)+.
The S-isomer could be obtained from (S)-((S)-1-phenylethyl) 2-
(dimethylamino)-2-(2-fluorophenyl)acetate TFA salt in similar fashion.
Cap-39
CI NH2 CI N
0 0 OH ________ ,..- 40 0 OH
cap-39
A mixture of (R)-(2-chlorophenyl)glycine (0.300 g, 1.62 mmol),
formaldehyde (35% aqueous solution, 0.80 mL, 3.23 mmol) and 20% Pd(OH)2/C
(0.050 g) was hydrogenated at room temperature and atmospheric pressure (F12
balloon) for 4 hours. The solution was then purged with Ar, filtered through
diatomaceous earth (Celite ) and concentrated in vacuo. The residue was
purified by
reverse-phase preparative HPLC (Primesphere C-18, 30 x 100mm; CH3CN-H20-
0.1% TFA) to give theTFA salt of the title compound (R)-2-(dimethylamino)-2-(2-
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chlorophenyl)acetic acid as a colorless oil (0.290 g, 55%). 1H NMR (400 MHz,
CD30D) 6 7.59-7.65 (m, 2H), 7.45-7.53 (m, 2H), 5.40 (s, 1H), 2.87 (s, 6H).
LCMS:
Anal. Calcd. for C10H12C1NO2: 213; found: 214 (M+H)+.
Cap-40
I
CI NH2 CI HN 0'
0 0 OH ___ . 0 0 OH
cap-40
To an ice-cold solution of (R)-(2-chlorophenyl)glycine (1.00 g, 5.38 mmol)
and NaOH (0.862 g, 21.6 mmol) in H20 (5.5 mL) was added methyl chloroformate
(1.00 mL, 13.5 mmol) dropwise. The mixture was allowed to stir at 0 C for 1
hour
and then it was acidified by the addition of conc. HC1 (2.5 mL). The mixture
was
extracted with ethyl acetate (2x) and the combined organic phase was washed
(H20,
brine), dried (Na2SO4), filtered, and concentrated in vacuo to give the title
compound
(R)-2-(methoxycarbonylamino)-2-(2-chlorophenyl)acetic acid as a yellow-orange
foam (1.31 g, 96%). 1FINMR (400 MHz, CD30D) 6 7.39 ¨ 7.43 (m, 2H), 7.29 ¨
7.31 (m, 2H), 5.69 (s, 1H), 3.65 (s, 3H). LCMS: Anal. Calcd. for C10H10C1N04:
243; found: 244 (M+H)+.
Cap-41
(-0
CINJ
0 0
= ____________________________ OH ______ .- .
OH
cap-41
To a suspension of 2-(2-(chloromethyl)phenyl)acetic acid (2.00 g, 10.8 mmol)
in THF (20 mL) was added morpholine (1.89 g, 21.7 mmol) and the solution was
stirred at room temperature for 3 hours. The reaction mixture was then diluted
with
ethyl acetate and extracted with H20 (2x). The aqueous phase was lyophilized
and
the residue was purified by silica gel chromatography (Biotage/ 0-10% methanol-
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CH202) to give the title compound 2-(2-(Morpholinomethyl)phenyl)acetic acid as
a
colorless solid (2.22 g, 87%). 1H NMR (400 MHz, CD30D) 6 7.37-7.44 (m, 3H),
7.29-7.33 (m, 1H), 4.24 (s, 2H), 3.83 (br s, 4H), 3.68 (s, 2H), 3.14 (br s,
4H). LCMS:
Anal. Calcd. for C13H17NO3: 235; found: 236 (M+H)+.
The following examples were similarly prepared using the method described
for Cap-41:
NO LCMS: Anal. Calcd. for
Cap-42 0 C14H19NO2: 233; found:
= OH 234 (M+H)+.
0 LCMS: Anal. Calcd. for
Cap-43 0 C13H17NO2: 219; found:
110 OH 220
Me
\
Nme LCMS: Anal. Calcd. for
--
Cap-44 0 C11H15NO2: 193; found:
0 OH 194
r re
LCMS: Anal. Calcd. for
N---/
Cap-45 0 C14H20N202: 248; found:
110 OH 249 (M+H)+.
Cap-45a
40 OH 40 OH
0 _______________________________________________ . 0
NH2 HNO
= pTs0H salt r
NH
Cap-45a
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HMDS (1.85 mL, 8.77 mmol) was added to a suspension of (R)-2-amino-2-
phenylacetic acid p-toluenesulfonate (2.83 g, 8.77 mmol) in CH2C12 (10 mL) and
the
mixture was stirred at room temperature for 30 minutes. Methyl isocyanate (0.5
g,
8.77 mmol) was added in one portion stirring continued for 30 minutes. The
reaction
was quenched by addition of H20 (5 mL) and the resulting precipitate was
filtered,
washed with H20 and n-hexanes, and dried under vacuum. (R)-2-(3-methylureido)-
2-phenylacetic acid (1.5 g; 82 %).was recovered as a white solid and it was
used
without further purification. 1H NMR (500 MHz, DMSO-d6) 6 ppm 2.54 (d, J=4.88
Hz, 3H) 5.17 (d, J=7.93 Hz, 1H) 5.95 (q, J=4.48 Hz, 1H) 6.66 (d, J=7.93 Hz,
1H)
7.26-7.38 (m, 5H) 12.67 (s, 1H). LCMS: Anal. Calcd. for C10H12N203 208.08
found 209.121 (M+H)+; HPLC Phenomenex C-18 3.0 x 46 mm, 0 to 100% B over 2
minutes, 1 minute hold time, A=90% water, 10% methanol, 0.1% TFA, B=10%
water, 90% methanol, 0.1% TFA, RT=1.38 min, 90% homogeneity index.
Cap-46
40 OH (00 OH
0 , 0
NH2 HNO
i
= pTs0H salt r NH
cap-46
The desired product was prepared according to the method described for Cap-
45a. 1H NMR (500 MHz, DMSO-d6) 6 ppm 0.96 (t, J=7.17 Hz, 3H) 2.94-3.05 (m,
2H) 5.17 (d, J=7.93 Hz, 1H) 6.05 (t, J=5.19 Hz, 1H) 6.60 (d, J=7.63 Hz, 1H)
7.26-
7.38 (m, 5H) 12.68 (s, 1H). LCMS: Anal. Calcd. for C11H14N203 222.10 found
223.15 (M+H)+. HPLC XTERRA C-18 3.0 x 506 mm, 0 to 100% B over 2 minutes,
1 minute hold time, A=90% water, 10% methanol, 0.2% H3PO4, B=10% water, 90%
methanol, 0.2% H3PO4, RT=0.87 min, 90% homogeneity index.
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Cap-47
0
, 01- 1 0 01- 2
__________________________________________________ . 01 OH
NH2 HN 0
i HN 0
i
A N
N
B Cap-47
Step 1; (R)-tert-butyl 2-(3,3-dimethylureido)-2-phenylacetate: To a stirred
solution of (R)-tert-butyl-2-amino-2-phenylacetate (1.0 g, 4.10 mmol) and
Hunig's
base (1.79 mL, 10.25 mmol) in DMF (40 mL) was added dimethylcarbamoyl
chloride (0.38 mL, 4.18 mmol) dropwise over 10 minutes. After stirring at room
temperature for 3 hours, the reaction was concentrated under reduced pressure
and
the resulting residue was dissolved in ethyl acetate. The organic layer was
washed
with H20, 1N aq. HC1 and brine, dried (MgSO4), filtered and concentrated under
reduced pressure. (R)-tert-butyl 2-(3,3-dimethylureido)-2-phenylacetate was
obtained as a white solid ( 0.86 g; 75%) and used without further
purification. 1H
NMR (500 MHz, DMSO-d6) 6 ppm 1.33 (s, 9H) 2.82 (s, 6H) 5.17 (d, J=7.63 Hz, 1H)
6.55 (d, J=7.32 Hz, 1H) 7.24-7.41 (m, 5H). LCMS: Anal. Calcd. for C15H22N203
278.16 found 279.23 (M+H)+; HPLC Phenomenex LUNA C-18 4.6 x 50 mm, 0 to
100% B over 4 minutes, 1 minute hold time, A=90% water, 10% methanol, 0.1%
TFA, B=10% water, 90% methanol, 0.1% TFA, RT=2.26 min, 97% homogeneity
index.
Step 2; (R)-2-(3,3-dimethylureido)-2-phenylacetic acid: To a stirred solution
of ((R)-tert-butyl 2-(3,3-dimethylureido)-2-phenylacetate ( 0.86 g, 3.10 mmol)
in
CH2C12 (250 mL) was added TFA (15 mL) dropwise and the resulting solution was
stirred at rt for 3 hours. The desired compound was then precipitated out of
solution
with a mixture of EtOAC:Hexanes (5:20), filtered off and dried under reduced
pressure. (R)-2-(3,3-dimethylureido)-2-phenylacetic acid was isolated as a
white
solid (0.59g, 86%) and used without further purification. 1H NMR (500 MHz,
DMSO-d6) 6 ppm 2.82 (s, 6H) 5.22 (d, J=7.32 Hz, 1H) 6.58 (d, J=7.32 Hz, 1H)
7.28
(t, J=7.17 Hz, 1H) 7.33 (t, J=7.32 Hz, 2H) 7.38-7.43 (m, 2H) 12.65 (s, 1H).
LCMS:
Anal. Calcd. for C11H14N203: 222.24; found: 223.21 (M+H)+. HPLC XTERRA C-
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18 3.0 x 50 mm, 0 to 100% B over 2 minutes, 1 minute hold time, A=90% water,
10% methanol, 0.2% H3PO4, B=10% water, 90% methanol, 0.2% H3PO4, RT=0.75
min, 93% homogeneity index.
Cap-48
O' 1 1 = 01- 2 40 OH
0 , 0 _ 0
nH2 HNO Hn
A crNH cr NH
Cap-48
Step 1; (R)-tert-butyl 2-(3-cyclopentylureido)-2-phenylacetate: To a stirred
solution of (R)-2-amino-2-phenylacetic acid hydrochloride (1.0 g, 4.10 mmol)
and
Hunig's base (1.0 mL, 6.15 mmol) in DMF (15 mL) was added cyclopentyl
isocyanate (0.46 mL, 4.10 mmol) dropwise and over 10 minutes. After stirring
at
room temperature for 3 hours, the reaction was concentrated under reduced
pressure
and the resulting residue was traken up in ethyl acetate. The organic layer
was
washed with H20 and brine, dried (MgSO4), filtered, and concentrated under
reduced
pressure. (R)-tert-butyl 2-(3-cyclopentylureido)-2-phenylacetate was obtained
as an
opaque oil (1.32 g; 100 %) and used without further purification. 1H NMR (500
MHz, CD3C1-D) 6 ppm 1.50-1.57 (m, 2H) 1.58-1.66 (m, 2H) 1.87-1.97 (m, 2H) 3.89-
3.98 (m, 1H) 5.37 (s, 1H) 7.26-7.38 (m, 5H). LCMS: Anal. Calcd. for C18H26N203
318.19 found 319.21 (M+H)+; HPLC XTERRA C-18 3.0 x 50 mm, 0 to 100% B
over 4 minutes, 1 minute hold time, A=90% water, 10% methanol, 0.1% TFA,
B=10% water, 90% methanol, 0.1% TFA, RT=2.82 min, 96% homogeneity index.
Step 2; (R)-2-(3-cyclopentylureido)-2-phenylacetic acid: To a stirred solution
of (R)-tert-butyl 2-(3-cyclopentylureido)-2-phenylacetate (1.31 g, 4.10 mmol)
in
CH2C12 (25 mL) was added TFA (4 mL) and trietheylsilane (1.64 mL; 10.3 mmol)
dropwise, and the resulting solution was stirred at room temperature for 6
hours. The
volatile components were removed under reduced pressure and the crude product
was
recrystallized in ethyl acetate/pentanes to yield (R)-2-(3-cyclopentylureido)-
2-
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phenylacetic acid as a white solid (0.69 g, 64%). 1H NMR (500 MHz, DMSO-d6) 6
ppm 1.17-1.35 (m, 2H) 1.42-1.52 (m, 2H) 1.53-1.64 (m, 2H) 1.67-1.80 (m, 2H)
3.75-
3.89 (m, 1H) 5.17 (d, J=7.93 Hz, 1H) 6.12 (d, J=7.32 Hz, 1H) 6.48 (d, J=7.93
Hz,
1H) 7.24-7.40 (m, 5H) 12.73 (s, 1H). LCMS: Anal. Calcd. for C14H18N203:
262.31;
found: 263.15 (M+H)+. HPLC XTERRA C-18 3.0 x 50 mm, 0 to 100% B over 2
minutes, 1 minute hold time, A=90% water, 10% methanol, 0.2% H3PO4, B=10%
water, 90% methanol, 0.2% H3PO4, RT=1.24 mm, 100% homogeneity index.
Cap-49
is NrOH s N OH
cap-49
To a stirred solution of 2-(benzylamino)acetic acid (2.0 g, 12.1 mmol) in
formic acid (91 mL) was added formaldehyde (6.94 mL, 93.2 mmol). After five
hours at 70 C, the reaction mixture was concentrated under reduced pressure
to 20
mL and a white solid precipitated. Following filtration, the mother liquors
were
collected and further concentrated under reduced pressure providing the crude
product. Purification by reverse-phase preparative HPLC (Xterra 30 X 100 mm,
detection at 220 nm, flow rate 35 mL/min, 0 to 35% B over 8 mm; A= 90% water,
10
% methanol, 0.1% TFA, B=10% water, 90 % methanol, 0.1% TFA) provided the title
compound 2-(benzyl(methyl)-amino)acetic acid as its TFA salt (723 mg, 33%) as
a
colorless wax. 1H NMR (300 MHz, DMSO-d6) 6 ppm 2.75 (s, 3H) 4.04 (s, 2H) 4.34
(s, 2H) 7.29-7.68 (m, 5H). LCMS: Anal. Calcd. for: C10H13NO2 179.09; Found:
180.20 (M+H)+.
Cap-50
\/
HN NO OH
I 0
101 Nil 0
Cap-50
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To a stirred solution of 3-methyl-2-(methylamino)butanoic acid (0.50 g, 3.81
mmol) in water ( 30 mL) was added K2CO3 (2.63 g, 19.1 mmol) and benzyl
chloride
(1.32 g, 11.4 mmol). The reaction mixture was stirred at ambient temperature
for 18
hours. The reaction mixture was extracted with ethyl acetate (30 mL x 2) and
the
aqueous layer was concentrated under reduced pressure providing the crude
product
which was purified by reverse-phase preparative HPLC (Xterra 30 x 100mm,
detection at 220 nm, flow rate 40 mL/min, 20 to 80% B over 6 min; A= 90%
water,
10% methanol, 0.1% TFA, B=10% water, 90% methanol, 0.1% TFA) to provide 2-
(benzyl(methyl)amino)-3-methylbutanoic acid, TFA salt (126 mg, 19%) as a
colorless wax. 1H NMR (500 MHz, DMSO-d6) 6 ppm 0.98 (d, 3H) 1.07 (d, 3H)
2.33-2.48 (m, 1H) 2.54-2.78 (m, 3H) 3.69 (s, 1H) 4.24 (s, 2H) 7.29-7.65 (m,
5H).
LCMS: Anal. Calcd. for: C13H19NO2 221.14; Found: 222.28 (M+H)+.
Cap-51
I
y 0
HNõ.AOH
,......--......
Na2CO3 (1.83g, 17.2 mmol) was added to NaOH (33 mL of 1M/H20, 33
mmol) solution of L-valine (3.9 g, 33.29 mmol) and the resulting solution was
cooled
with ice-water bath. Methyl chloroformate (2.8 mL, 36.1 mmol) was added
dropwise
over 15 min, the cooling bath was removed and the reaction mixture was stirred
at
ambient temperature for 3.25 hr. The reaction mixture was washed with ether
(50
mL, 3x), and the aqueous phase was cooled with ice-water bath and acidified
with
concentrated HC1 to a pH region of 1-2, and extracted with CH2C12 (50 mL, 3x).
The
organic phase was dried (MgSO4) and evaporated in vacuo to afford Cap-51 as a
white solid (6 g). 1H NMR for the dominant rotamer (DMSO-d6, cF2.5 ppm, 500
MHz): 12.54 (s, 1H), 7.33 (d, J=8.6, 1H), 3.84 (dd, J=8.4, 6.0, 1H), 3.54 (s,
3H),
2.03 (m, 1H), 0.87 (m, 6H). HRMS: Anal. Calcd. for [M+H]+ C7H14N04: 176.0923;
found 176.0922.
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Cap 51 (alternate route)
o/ o/
H2N HNO HNio
o 0
0-tBu--"S \
0-tBu OH
DIEA (137.5 mL, 0.766 mol) was added to a suspension of (S)-tert-butyl
amino-3-methylbutanoate hydrochloride (75.0 g, 0.357 mol) in THF (900 mL), and
the
mixture was cooled to 0 C (ice/water bath). Methyl chloroformate (29.0 mL,
0.375
mol) was added dropwise over 45 min, the cooling bath was removed and the
heterogeneous mixture was stirred at ambient temperature for 3 h. The solvent
was
removed under diminished pressure and the residue partitioned between Et0Ac
and
water (1 L each). The organic layer was washed with H20 (1 L) and brine (1 L),
dried
(MgSO4), filtered and concentrated under diminished pressure. The crude
material was
passed through a plug of silica gel (1 kg), eluting with hexanes (4 L) and
15:85
Et0Ac/hexanes (4 L) to afford (S)-tert-butyl 2-(methoxycarbonylamino)-3-
methylbutanoate as a clear oil (82.0 g, 99% yield). 1H-NMR (500 MHz, DMSO-d6,
6
= 2.5 ppm) 7.34 (d, J= 8.6, 1 H), 3.77 (dd, J= 8.6, 6.1, 1 H), 3.53 (s, 3 H),
1.94 -2.05
(m, 1 H), 1.39 (s, 9 H), 0.83 - 0.92 (m, 6 H). 13C-NMR (126 MHz, DMSO-d6, 6 =
39.2
ppm) 170.92, 156.84, 80.38, 60.00, 51.34, 29.76, 27.62, 18.92, 17.95. LC/MS:
[M+Na]+ 254.17.
Trifluoroacetic acid (343 mL, 4.62 mol) and Et3SiH (142 mL, 0.887 mol) were
added sequentially to a solution of (S)-tert-butyl 2-(methoxycarbonylamino)-3-
methylbutanoate (82.0 g, 0.355 mol) in CH2C12 (675 mL), and the mixture was
stirred
at ambient temperature for 4 h. The volatile component was removed under
diminished pressure and the resultant oil triturated with petroleum ether (600
mL) to
afford a white solid, which was filtered and washed with hexanes (500 mL) and
petroleum ether (500 mL). Recrystallization from Et0Ac/petroleum ether
afforded
Cap-51 as white flaky crystals (54.8 g, 88 % yield). MP = 108.5-109.5 C. 1H
NMR
(500 MHz, DMSO-d6, 6 = 2.5 ppm) 12.52 (s, 1 H), 7.31 (d, J= 8.6, 1 H), 3.83
(dd, J=
8.6, 6.1, 1 H), 3.53 (s, 3 H), 1.94 - 2.07 (m, 1 H), 0.86 (dd, J= 8.9, 7.0, 6
H). 13C NMR
(126 MHz, DMSO-d6, 6 = 39.2 ppm) 173.30, 156.94, 59.48, 51.37, 29.52, 19.15,
17.98.
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LC/MS: [M+H]+ = 176.11. Anal. Calcd. for C7F113N04: C, 47.99; H, 7.48; N,
7.99.
Found: C, 48.17; H, 7.55; N, 7.99. Optical Rotation: [a] D = -4.16 (12.02
mg/mL;
Me0H). Optical purity: >99.5 % ee. Note: the optical purity assessment was
made on
the methyl ester derivative of Cap-51, which was prepared under a standard
TMSCHN2
(benzene/Me0H) esterification protocol. HPLC analytical conditions: column,
ChiralPak AD-H (4.6 x 250mm, 51.im); solvent, 95% heptane / 5% IPA
(isocratic); flow
rate, 1 mL/min; temperature, 35 C; UV monitored at 205 nm.
[Note: Cap 51 could also be purchased from Flamm.]
Cap-52 (Same as Cap-12)
I
y 0
HN õ,
)LOH
Cap-52 was synthesized from L-alanine according to the procedure described
for the synthesis of Cap-51. For characterization purposes, a portion of the
crude
material was purified by a reverse phase HPLC (H20/methanol/TFA) to afford Cap-
52 as a colorless viscous oil. 1H NMR (DMSO-d6, cF2.5 ppm, 500 MHz): 12.49 (br
s, 1H), 7.43 (d, J=7.3, 0.88H), 7.09 (app br s, 0.12H), 3.97 (m, 1H), 3.53 (s,
3H),
1.25 (d, J=7.3, 3H).
Cap-53 to -64 were prepared from appropriate starting materials according to
the procedure described for the synthesis of Cap-51, with noted modifications
if any.
Cap Structure Data
Cap-53a: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
(R) MHz): 6 12.51 (br s, 1H), 7.4 (d, J= 7.9,
Cap-53b: 0.9H), 7.06 (app s, 0.1H), 3.86-3.82 (m,
14 0
(S) 0y ki)-LOH 1H), 3.53 (s, 3H), 1.75-1.67 (m, 1H),
1.62-1.54 (m, 1H), 0.88 (d, J= 7.3, 3H).
0
RT = 0.77 minutes (Cond. 2); LC/MS:
Anal. Calcd. for [M+Na]+ C6K1NNaa4 :
184.06; found 184.07. HRMS Calcd. for
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[M+Na]+ C6K1NNa04: 184.0586; found
184.0592.
Cap-54a: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
(R) MHz): 6 12.48 (s, 1H), 7.58 (d, J = 7.6,
Cap-54b: 0.9H), 7.25 (app s, 0.1H), 3.52 (s, 3H),
0
(S) 0y NHIAOH 3.36-3.33 (m, 1H), 1.10-1.01 (m, 1H),
0 0.54-0.49 (m, 1H), 0.46-0.40 (m, 1H),
0.39-0.35 (m, 1H), 0.31-0.21 (m, 1H).
HRMS Calcd. for [M+H]+ C7H12N04:
174.0766; found 174.0771
Cap-55 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
MHz): 6 12.62 (s, 1H), 7.42 (d, J = 8.2,
0
O H,A 0.9H), 7.07 (app s, 0.1H), 5.80-
5.72 (m,
y . OH
1H), 5.10 (d, J = 17.1, 1H), 5.04 (d, J =
0
II 10.4, 1H), 4.01-3.96 (m, 1H), 3.53 (s,
3H), 2.47-2.42 (m, 1H), 2.35-2.29 (m,
1H).
Cap-56 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
0
O H,A MHz): 6 12.75 (s, 1H), 7.38 (d, J
= 8.3,
II , . OH
0.9H), 6.96 (app s, 0.1H), 4.20-4.16 (m,
0 -0
I 1H), 3.60-3.55 (m, 2H), 3.54 (s, 3H),
3.24
(s, 3H).
Cap-57 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
MHz): 6 12.50 (s, 1H), 8.02 (d, J = 7.7,
0.08H), 7.40 (d, J= 7.9, 0.76H), 7.19 (d,
0
O H,A J = 8.2, 0.07H), 7.07 (d, J = 6.7,
0.09H),
y . OH
4.21-4.12 (m, 0.08H), 4.06-3.97 (m,
0 )0.07H), 3.96-3.80 (m, 0.85H), 3.53 (s,
3H), 1.69-1.51 (m, 2H), 1.39-1.26 (m,
2H), 0.85 (t, J = 7.4, 3H). LC (Cond. 2):
RT = 1.39 LC/MS: Anal. Calcd. for
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[M+H]+ C7H14N04: 176.09; found
176.06.
Cap-58 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
MHz): 6 12.63 (br s, 1H), 7.35 (s,1H),
O 7.31 ( d, J = 8.2, 1H), 6.92 (s, 1H), 4.33-
O ENIõ A
y = OH 4.29 (m, 1H), 3.54 (s, 3H), 2.54(dd, J =
0 NH2 15.5, 5.4, 1H), 2.43 (dd, J = 15.6, 8.0,
O 1H). RT = 0.16 min (Cond. 2); LC/MS:
Anal. Calcd. for [M+H]+ C6H11N205:
191.07; found 191.14.
Cap-59a: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400
(R) MHz): 6 12.49 (br s, 1H), 7.40 (d, J = 7.3,
Cap-59b: 0 0.89H), 7.04 (br s, 0.11H), 4.00-3.95 (m,
H
(S) OyN).LOH 3H), 1.24 (d, J= 7.3, 3H), 1.15 (t, J =
0 7.2, 3H). HRMS: Anal. Calcd. for
[M+H]+ C6H12N04: 162.0766; found
162.0771.
Cap-60 The crude material was purified with a
reverse phase HPLC (H20/Me0H/TFA)
to afford a colorless viscous oil that
crystallized to a white solid upon
O exposure to high vacuum. 1H NMR
H.L
O N
y OH (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 6
0 12.38 (br s, 1H), 7.74 (s, 0.82H), 7.48
(s,
0.18H), 3.54/3.51 (two s, 3H), 1.30 (m,
2H), 0.98 (m, 2H). HRMS: Anal. Calcd.
for [M+H]+ C6H10N04: 160.0610; found
160.0604.
Cap-61 0 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400
H*L
O N
y OH MHz): 6 12.27 (br s, 1H), 7.40 (br s,
1H),
0
3.50 (s, 3H), 1.32 (s, 6H). HRMS: Anal.
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Calcd. for [M+H]+ C6H12N04: 162.0766;
found 162.0765.
Cap-62 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400
MHz): 6 12.74 (br s, 1H), 4.21 (d, J =
10.3, 0.6H), 4.05 (d, J = 10.0, 0.4H),
1 0
0 N I=L 3.62/3.60 (two singlets, 3H), 3.0 (s,
3H),
0 2.14-2.05 (m, 1H), 0.95 (d, J = 6.3, 3H),
0.81 (d, J = 6.6, 3H). LC/MS: Anal.
Calcd. for [M-H] C8H14N04: 188.09;
found 188.05.
Cap-63 [Note: the reaction was allowed to run for
longer than what was noted for the
general procedure.] 1H NMR (DMSO-d6,
0
H.L
0yN OH 6 = 2.5 ppm, 400 MHz): 12.21 (br s, 1H),
0 7.42 (br s, 1H), 3.50 (s, 3H), 2.02-1.85
(m, 4H), 1.66-1.58 (m, 4H). LC/MS:
Anal. Calcd. for [M+H]+ C8H14N04:
188.09; found 188.19.
Cap-64 [Note: the reaction was allowed to run for
longer than what was noted for the
0 general procedure.] 1H NMR (DMSO-d6,
HeL
0 N
y OH 6 = 2.5 ppm, 400 MHz): 12.35 (br s, 1H),
0
7.77 (s, 0.82H), 7.56/7.52 (overlapping br
s, 0.18H), 3.50 (s, 3H), 2.47-2.40 (m, 2H),
2.14-2.07 (m, 2H), 1.93-1.82 (m, 2H).
Cap-65
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0
0
kl,'' A
y OH
0
- \ OH
Methyl chloroformate (0.65 mL, 8.39 mmol) was added dropwise over 5 min
to a cooled (ice-water) mixture of Na2CO3 (0.449 g, 4.23 mmol), NaOH (8.2 mL
of
1M/H20, 8.2 mmol) and (S)-2-amino-3-hydroxy-3-methylbutanoic acid (1.04 g,
7.81
mmol). The reaction mixture was stirred for 45 min, and then the cooling bath
was
removed and stirring was continued for an additional 3.75 hr. The reaction
mixture
was washed with CH2C12, and the aqueous phase was cooled with ice-water bath
and
acidified with concentrated HC1 to a pH region of 1-2. The volatile component
was
removed in vacuo and the residue was taken up in a 2:1 mixture of Me0H/CH2C12
(15 mL) and filtered, and the filterate was rotervaped to afford Cap-65 as a
white
semi-viscous foam (1.236 g). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 6 6.94
(d, J = 8.5, 0.9 H), 6.53 (br s, 0.1H), 3.89 (d, J = 8.8, 1H), 2.94 (s, 3H),
1.15 (s, 3H),
1.13 (s, 3H).
Cap-66 and -67 were prepared from appropriate commercially available
starting materials by employing the procedure described for the synthesis of
Cap-65.
Cap-66
0
0 t\l','' A
y OH
ID
1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 6 12.58 (br s, 1H), 7.07 (d, J =
8.3, 0.13H), 6.81 (d, J= 8.8, 0.67H), 4.10-4.02 (m, 1.15H), 3.91 (dd, J= 9.1,
3.5,
0.85H), 3.56 (s, 3H), 1.09 (d, J = 6.2, 3H). [Note: only the dominant signals
of NH
were noted].
Cap-67
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0
0 EN',A
y ' OH
ID OH
1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 12.51 (br s, 1H), 7.25 (d, J =
8.4, 0.75H), 7.12 (br d, J = 0.4, 0.05H), 6.86 (br s, 0.08H), 3.95-3.85 (m,
2H), 3.54
(s, 3H), 1.08 (d, J = 6.3, 3H). [Note: only the dominant signals of NH were
noted].
Cap-68
0
0 N
0 -r0
0,6n
Methyl chloroformate (0.38 ml, 4.9 mmol) was added drop-wise to a mixture
of 1N NaOH (aq) (9.0 ml, 9.0 mmol), 1M NaHCO3 (aq) (9.0 ml, 9.0 mol), L-
aspartic
acid 13-benzyl ester (1.0 g, 4.5 mmol) and Dioxane (9 m1). The reaction
mixture was
stirred at ambient conditions for 3 hr, and then washed with Ethyl acetate (50
ml, 3x).
The aqueous layer was acidified with 12N HC1to a pH ¨ 1-2, and extracted with
ethyl acetate (3 x 50 m1). The combined organic layers were washed with brine,
dried (Na2SO4), filtered, and concentrated in vacuo to afford Cap-68 as a
light yellow
oil (1.37g; mass is above theoretical yield, and the product was used without
further
purification). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500 MHz): 6 12.88 (br s, 1H),
7.55
(d, J = 8.5, 1H), 7.40-7.32 (m, 5H), 5.13 (d, J = 12.8, 1H), 5.10 (d, J =
12.9, 1H),
4.42-4.38 (m, 1H), 3.55 (s, 3H), 2.87 (dd, J = 16.2, 5.5, 1H), 2.71 (dd, J
=16.2, 8.3,
1H). LC (Cond. 2): RT = 1.90 min; LC/MS: Anal. Calcd. For [M+H]+ C13H16N06:
282.10; found 282.12.
Cap-69a and -69b
---1 0
Cap-69a: (R)-enantiomer
Cap-69b: (S)-enantiomer
NaCNBH3 (2.416 g, 36.5 mmol) was added in batches to a chilled (-15 C)
water (17 mL)/Me0H (10 mL) solution of alanine (1.338 g, 15.0 mmol). A few
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minutes later acetaldehyde (4.0 mL, 71.3 mmol) was added drop-wise over 4 min,
the
cooling bath was removed, and the reaction mixture was stirred at ambient
condition
for 6 hr. An additional acetaldehyde (4.0 mL) was added and the reaction was
stirred
for 2 hr. Concentrated HC1 was added slowly to the reaction mixture until the
pH
reached ¨ 1.5, and the resulting mixture was heated for 1 hr at 40 C. Most of
the
volatile component was removed in vacuo and the residue was purified with a
Dowex0 50WX8-100 ion-exchange resin (column was washed with water, and the
compound was eluted with dilute NH4OH, prepared by mixing 18 ml of NH4OH and
282 ml of water) to afford Cap-69 (2.0 g) as an off-white soft hygroscopic
solid. 1H
NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 6 3.44 (q, J = 7.1, 1H), 2.99-2.90 (m,
2H), 2.89-2.80 (m, 2H), 1.23 (d, J = 7.1, 3H), 1.13 (t, J = 7.3, 6H).
Cap-70 to -74x were prepared according to the procedure described for the
synthesis of Cap-69 by employing appropriate starting materials.
Cap-70a: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400
(R) MHz): 6 3.42 (q, J = 7.1, 1H), 2.68-2.60
Cap-70b: (S) 0 (m, 4H), 1.53-1.44(m, 4H), 1.19 (d, J =
N.).LOH 7.3, 3H), 0.85 (t, J = 7.5, 6H). LC/MS:
Anal. Calcd. for [M+H]+ C9H20NO2:
174.15; found 174.13.
Cap-71a: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
(R) MHz): 6 3.18-3.14 (m, 1H), 2.84-2.77
Cap-71b: (S) ------\ 0 (m, 2H), 2.76-2.68 (m, 2H), 1.69-
1.54
\--N )*LOH (m, 2H), 1.05 (t, J = 7.2, 6H), 0.91 (t,
J
/ = 7.3, 3H). LC/MS: Anal. Calcd. for
[M+H]+ CsHisNO2: 160.13; found
160.06.
Cap-72 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400
----1 0 MHz): 6 2.77-2.66 (m, 3H), 2.39-2.31
N).LOH (m, 2H), 1.94-1.85 (m, 1H), 0.98 (t, J =
7.1, 6H), 0.91 (d, J= 6.5, 3H), 0.85 (d, J
= 6.5, 3H). LC/MS: Anal. Calcd. for
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[M+H]+ C9H20NO2: 174.15; found
174.15.
Cap-73 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
0 MHz): 6 9.5 (br s, 1H), 3.77 (dd, J =
Nj-
. OH 10.8, 4.1,1H), 3.69-3.61 ( m, 2H), 3.26
0 (s, 3H), 2.99-2.88 (m, 4H), 1.13 (t, J =
1
7.2, 6H).
Cap-74 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500
MHz): 6 7.54 (s, 1H), 6.89 (s, 1H), 3.81
----\ 0 (t, J = 6.6, k,1H), 2.82-2.71 (m, 4H),
N
OH 2.63 (dd, J = 15.6, 7.0, 1H), 2.36 (dd, J
0
= 15.4, 6.3, 1H), 1.09 (t, J = 7.2, 6H).
NH2 RT = 0.125 minutes (Cond. 2); LC/MS:
Anal. Calcd. for [M+H]+ C81-112N203:
189.12; found 189.13.
Cap-74x "Th 0
\NLOH LC/MS: Anal. Calcd. for [M+H]+
C10H22NO2: 188.17; found 188.21
Cap-75
0
NJLOH
0
1
Cap-75, step a
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----\ 0
,Bn
0
OH
NaBH3CN (1.6 g, 25.5 mmol) was added to a cooled (ice/water bath) water
(25 ml)/methanol (15 ml) solution of H-D-Ser-OBz1 HC1 (2.0 g, 8.6 mmol).
Acetaldehyde (1.5 ml, 12.5 mmol) was added drop-wise over 5 min, the cooling
bath
was removed, and the reaction mixture was stirred at ambient condition for 2
hr. The
reaction was carefully quenched with 12N HC1 and concentrated in vacuo. The
residue was dissolved in water and purified with a reverse phase HPLC
(Me0H/H20/TFA) to afford the TFA salt of (R)-benzyl 2-(diethylamino)-3-
hydroxypropanoate as a colorless viscous oil (1.9g). 1H NMR (DMSO-d6, 6 = 2.5
ppm, 500 MHz): 6 9.73 (br s, 1H), 7.52-7.36 (m, 5H), 5.32 (d, J = 12.2, 1H),
5.27 (d,
J= 12.5, 1H), 4.54-4.32 (m, 1H), 4.05-3.97 (m, 2H), 3.43-3.21 (m, 4H), 1.23
(t, J=
7.2, 6H). LC/MS (Cond. 2): RT = 1.38 min; LC/MS: Anal. Calcd. for [M+H]+
C14H22NO3: 252.16; found 252.19.
Cap-75
NaH (0.0727 g, 1.82 mmol, 60%) was added to a cooled (ice-water) THF (3.0
mL) solution of the TFA salt (R)-benzyl 2-(diethylamino)-3-hydroxypropanoate
(0.3019 g, 0.8264 mmol) prepared above, and the mixture was stirred for 15
min.
Methyl iodide (56 L, 0.90 mmol) was added and stirring was continued for 18
hr
while allowing the bath to thaw to ambient condition. The reaction was
quenched
with water and loaded onto a Me0H pre-conditioned MCX (6 g) cartridge, and
washed with methanol followed by compound elution with 2N NH3/Methanol.
Removal of the volatile component in vacuo afforded Cap-75, contaminated with
(R)-2-(diethylamino)-3-hydroxypropanoic acid, as a yellow semi-solid (100 mg).
The product was used as is without further purification.
Cap-76
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H j?
0
N
c
NaCNBH3 (1.60 g, 24.2 mmol) was added in batches to a chilled (-15 C)
water/Me0H (12 mL each) solution of (S)-4-amino-2-(tert-butoxycarbonylamino)
butanoic acid (2.17 g, 9.94 mmol). A few minutes later acetaldehyde (2.7 mL,
48.1
mmol) was added drop-wise over 2 min, the cooling bath was removed, and the
reaction mixture was stirred at ambient condition for 3.5 hr. An additional
acetaldehyde (2.7 mL, 48.1 mmol) was added and the reaction was stirred for
20.5 hr.
Most of the Me0H component was removed in vacuo, and the remaining mixture
was treated with concentrated HC1 until its pH reached ¨ 1.0 and then heated
for 2 hr
at 40 C. The volatile component was removed in vacuo, and the residue was
treated
with 4 M HC1/dioxane (20 mL) and stirred at ambient condition for 7.5 hr. The
volatile component was removed in vacuo and the residue was purified with
Dowex
0 50WX8-100 ion-exchange resin (column was washed with water and the
compound was eluted with dilute NH4OH, prepared from 18 ml of NH4OH and 282
ml of water) to afford intermediate (S)-2-amino-4-(diethylamino)butanoic acid
as an
off-white solid (1.73 g).
Methyl chloroformate (0.36 mL, 4.65 mmol) was added drop-wise over 11
min to a cooled (ice-water) mixture of Na2CO3 (0.243 g, 2.29 mmol), NaOH (4.6
mL
of 1M/H20, 4.6 mmol) and the above product (802.4 mg). The reaction mixture
was
stirred for 55 min, and then the cooling bath was removed and stirring was
continued
for an additional 5.25 hr. The reaction mixture was diluted with equal volume
of
water and washed with CH2C12 (30 mL, 2x), and the aqueous phase was cooled
with
ice-water bath and acidified with concentrated HC1 to a pH region of 2. The
volatile
component was then removed in vacuo and the crude material was free-based with
MCX resin (6.0g; column was washed with water, and sample was eluted with 2.0
M
NH3/Me0H) to afford impure Cap-76 as an off-white solid (704 mg). 1H NMR
(Me0H-d4, 6 = 3.29 ppm, 400 MHz): 6 3.99 (dd, J = 7.5, 4.7, 1H), 3.62 (s, 3H),
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3.25-3.06 (m, 6H), 2.18-2.09 (m, 1H), 2.04-1.96 (m, 1H), 1.28 (t, J= 7.3, 6H).
LC/MS: Anal. Calcd. for [M+H]+ C10H21N204: 233.15; found 233.24.
Cap-77a and -77b
So
OH
N
Cap-77a: enantiomer-1
Cap-77b: enantiomer-2
The synthesis of Cap-77 was conducted according to the procedure described
for Cap-7 by using 7-azabicyclo[2.2.1]heptane for the SN2 displacement step,
and by
effecting the enantiomeric separation of the intermediate benzyl 2-(7-
azabicyclo[2.2.1]heptan-7-y1)-2-phenylacetate using the following condition:
the
intermediate (303.7 mg) was dissolved in ethanol, and the resulting solution
was
injected on a chiral HPLC column (Chiracel AD-H column, 30 x 250 mm, 5 um)
eluting with 90% CO2-10% Et0H at 70 mL/min, and a temperature of 35 C to
provide 124.5 mg of enantiomer-1 and 133.8 mg of enantiomer-2. These benzyl
esters were hydrogenolysed according to the preparation of Cap-7 to provide
Cap-77:
1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 6 7.55 (m, 2H), 7.38-7.30 (m, 3H),
4.16 (s, 1H), 3.54 (app br s, 2H), 2.08-1.88 (m, 4 H), 1.57-1.46 (m, 4H). LC
(Cond.
1): RT = 0.67 min; LC/MS: Anal. Calcd. for [M+H]+ C14H18NO2: 232.13; found
232.18. HRMS: Anal. Calcd. for [M+H]+ C14H18NO2: 232.1338; found 232.1340.
Cap-78
SO
. OH
v N
NaCNBH3 (0.5828 g, 9.27 mmol) was added to a mixture of the HC1 salt of
(R)-2-(ethylamino)-2-phenylacetic acid (an intermediate in the synthesis of
Cap-3;
0.9923 mg, 4.60 mmol) and (1-ethoxycyclopropoxy)trimethylsilane (1.640 g, 9.40
mmol) in Me0H (10 mL), and the semi-heterogeneous mixture was heated at 50 C
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with an oil bath for 20 hr. More (1-ethoxycyclopropoxy)trimethylsilane (150
mg,
0.86 mmol) and NaCNBH3 (52 mg, 0.827 mmol) were added and the reaction
mixture was heated for an additional 3.5 hr. It was then allowed to cool to
ambient
temperature and acidified to a ¨ pH region of 2 with concentrated HC1, and the
mixture was filtered and the filtrate was rotervaped. The resulting crude
material was
taken up in i-PrOH (6 mL) and heated to effect dissolution, and the non-
dissolved
part was filtered off and the filtrate concentrated in vacuo. About 1/3 of the
resultant
crude material was purified with a reverse phase HPLC (H20/Me0H/TFA) to afford
the TFA salt of Cap-78 as a colorless viscous oil (353 mg). 1H NMR (DMSO-d6, 6
=
2.5 ppm, 400 MHz; after D20 exchange): 6 7.56-7.49 (m, 5H), 5.35 (S, 1H), 3.35
(m,
1H), 3.06 (app br s, 1H), 2.66 (m, 1H), 1.26 (t, J = 7.3, 3H), 0.92 (m, 1H),
0.83-0.44
(m, 3H). LC (Cond. 1): RT = 0.64 min; LC/MS: Anal. Calcd. for [M+H]+
C13H18NO2: 220.13; found 220.21. HRMS: Anal. Calcd. for [M+H]+ C13H18NO2:
220.1338; found 220.1343.
Cap-79
0
0
N
Co)
Ozone was bubbled through a cooled (-78 C) CH2C12 (5.0 mL) solution Cap-
55 (369 mg, 2.13 mmol) for about 50 min until the reaction mixture attained a
tint of
blue color. Me2S (10 pipet drops) was added, and the reaction mixture was
stirred
for 35 min. The -78 C bath was replaced with a -10 C bath and stirring
continued
for an additional 30 min, and then the volatile component was removed in vacuo
to
afford a colorless viscous oil.
NaBH3CN (149 mg, 2.25 mmol) was added to a Me0H (5.0 mL) solution of
the above crude material and morpholine (500 [IL, 5.72 mmol) and the mixture
was
stirred at ambient condition for 4 hr. It was cooled to ice-water temperature
and
treated with concentrated HC1 to bring its pH to ¨2.0, and then stirred for
2.5 hr. The
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volatile component was removed in vacuo, and the residue was purified with a
combination of MCX resin (Me0H wash; 2.0 N NH3/Me0H elution) and a reverse
phase HPLC (H20/Me0H/TFA) to afford Cap-79 containing unknown amount of
morpholine.
In order to consume the morpholine contaminant, the above material was
dissolved in CH2C12 (1.5 mL) and treated with Et3N (0.27 mL, 1.94 mmol)
followed
by acetic anhydride (0.10 mL, 1.06 mmol) and stirred at ambient condition for
18 hr.
THF (1.0 mL) and H20 (0.5 mL) were added and stirring continued for 1.5 hr.
The
volatile component was removed in vacuo, and the resultant residue was passed
through MCX resin (Me0H wash; 2.0 N NH3/Me0H elution) to afford impure Cap-
79 as a brown viscous oil, which was used for the next step without further
purification.
Cap-80a and -80b
0
0 N,
IIOH
Cap-80a: S/S-diastereomer
0 Cap-80b: S/R-diastereomer
0-si/ (
SOC12 (6.60 mL, 90.5 mmol) was added drop-wise over 15 min to a cooled
(ice-water) mixture of (S)-3-amino-4-(benzyloxy)-4-oxobutanoic acid (10.04g,
44.98
mmol) and Me0H (300 mL), the cooling bath was removed and the reaction mixture
was stirred at ambient condition for 29 hr. Most of the volatile component was
removed in vacuo and the residue was carefully partitioned between Et0Ac (150
mL)
and saturated NaHCO3 solution. The aqueous phase was extracted with Et0Ac (150
mL, 2x), and the combined organic phase was dried (MgSO4), filtered, and
concentrated in vacuo to afford (S)-1-benzyl 4-methyl 2-aminosuccinate as a
colorless oil (9.706g). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 6 7.40-7.32
(m,
5H), 5.11 (s, 2H), 3.72 (app t, J = 6.6, 1H), 3.55 (s, 3H), 2.68 (dd, J =
15.9, 6.3, 1H),
2.58 (dd, J = 15.9, 6.8, 1H), 1.96 (s, 2H). LC (Cond. 1): RT = 0.90 min;
LC/MS:
Anal. Calcd. for [M+H]+ C12H16N04: 238.11; found 238.22.
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Pb(NO3)2 (6.06 g, 18.3 mmol) was added over 1 min to a CH2C12 (80 mL)
solution of (S)-1-benzyl 4-methyl 2-aminosuccinate (4.50 g, 19.0 mmol), 9-
bromo-9-
pheny1-9H-fluorene (6.44 g, 20.0 mmol) and Et3N (3.0 mL, 21.5 mmol), and the
heterogeneous mixture was stirred at ambient condition for 48 hr. The mixture
was
filtered and the filtrate was treated with MgSO4 and filtered again, and the
final
filtrate was concentrated. The resulting crude material was submitted to a
Biotage
purification (350 g silica gel, CH2C12 elution) to afford (S)-1-benzyl 4-
methyl 2-(9-
pheny1-9H-fluoren-9-ylamino)succinate as highly viscous colorless oil (7.93
g). 1H
NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 6 7.82 (m, 2H), 7.39-7.13 (m, 16H), 4.71
(d, J = 12.4, 1H), 4.51 (d, J = 12.6, 1H), 3.78 (d, J = 9.1, NH), 3.50 (s,
3H), 2.99 (m,
1H), 2.50-2.41 (m, 2H, partially overlapped with solvent). LC (Cond. 1): RT =
2.16
min; LC/MS: Anal. Calcd. for [M+H]+ C31F128N04: 478.20; found 478.19.
LiHMDS (9.2 mL of 1.0 M/THF, 9.2 mmol) was added drop-wise over 10
min to a cooled (-78 C) THF (50 mL) solution of (S)-1-benzyl 4-methyl 2-(9-
phenyl-9H-fluoren-9-ylamino)succinate (3.907 g, 8.18 mmol) and stirred for -1
hr.
Mel (0.57 mL, 9.2 mmol) was added drop-wise over 8 min to the mixture, and
stirring was continued for 16.5 hr while allowing the cooling bath to thaw to
room
temperature. After quenching with saturated NH4C1 solution (5 mL), most of the
organic component was removed in vacuo and the residue was partitioned between
CH2C12 (100 mL) and water (40 mL). The organic layer was dried (MgSO4),
filtered,
and concentrated in vacuo, and the resulting crude material was purified with
a
Biotage (350 g silica gel; 25% Et0Ac/hexanes) to afford 3.65 g of a 2S/3S and
2S/3R
diastereomeric mixtures of 1-benzyl 4-methyl 3-methy1-2-(9-pheny1-9H-fluoren-9-
ylamino)succinate in -1.0:0.65 ratio (1H NMR). The stereochemistry of the
dominant isomer was not determined at this juncture, and the mixture was
submitted
to the next step without separation. Partial 1H NMR data (DMSO-d6, 6 = 2.5
ppm,
400 MHz): major diastereomer, 6 4.39 (d, J = 12.3, 1H of CH2), 3.33 (s, 3H,
overlapped with H20 signal), 3.50 (d, J = 10.9, NH), 1.13 (d, J = 7.1, 3H);
minor
diastereomer, 6 4.27 (d, J =12.3, 1H of CH2), 3.76 (d, J = 10.9, NH), 3.64 (s,
3H),
0.77 (d, J = 7.0, 3H). LC (Cond. 1): RT = 2.19 min; LC/MS: Anal. Calcd. for
[M+H]+ C32H30N04: 492.22; found 492.15.
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Diisobutylaluminum hydride (20.57 ml of 1.0 M in hexanes, 20.57 mmol)
was added drop-wise over 10 min to a cooled (-78 C) THF (120 mL) solution of
(2S)-1-benzyl 4-methyl 3-methy1-2-(9-pheny1-9H-fluoren-9-ylamino)succinate
(3.37
g, 6.86 mmol) prepared above, and stirred at -78 C for 20 hr. The reaction
mixture
was removed from the cooling bath and rapidly poured into ¨1M H3PO4/H20 (250
mL) with stirring, and the mixture was extracted with ether (100 mL, 2x). The
combined organic phase was washed with brine, dried (MgSO4), filtered and
concentrated in vacuo. A silica gel mesh of the crude material was prepared
and
submitted to chromatography (25% Et0Ac/hexanes; gravity elution) to afford
1.1g of
(2S,3S)-benzyl 4-hydroxy-3-methy1-2-(9-pheny1-9H-fluoren-9-ylamino)butanoate,
contaminated with benzyl alcohol, as a colorless viscous oil and (2S,3R)-
benzyl 4-
hydroxy-3-methy1-2-(9-pheny1-9H-fluoren-9-ylamino)butanoate containing the
(2S,3R) stereoisomer as an impurity. The later sample was resubmitted to the
same
column chromatography purification conditions to afford 750 mg of purified
material
as a white foam. [Note: the (2S, 3S) isomer elutes before the (2S,3R) isomer
under
the above condition]. (2S, 3S) isomer: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz):
7.81 (m, 2H), 7.39-7.08 (m, 16H), 4.67 (d, J = 12.3, 1H), 4.43 (d, J = 12.4,
1H), 4.21
(app t, J = 5.2, OH), 3.22 (d, J = 10.1, NH), 3.17 (m, 1H), 3.08 (m, 1H), ¨2.5
(m, 1H,
overlapped with the solvent signal), 1.58 (m, 1H), 0.88 (d, J = 6.8, 3H). LC
(Cond.
1): RT = 2.00 min; LC/MS: Anal. Calcd. for [M+H]+ C31H30NO3: 464.45; found
464.22. (2S, 3R) isomer: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz): 7.81 (d, J =
7.5, 2H), 7.39-7.10 (m, 16H), 4.63 (d, J= 12.1, 1H), 4.50 (app t, J = 4.9,
1H), 4.32 (d,
J= 12.1, 1H), 3.59-3.53 (m, 2H), 3.23 (m, 1H), 2.44 (dd, J = 9.0, 8.3, 1H),
1.70 (m,
1H), 0.57 (d, J = 6.8, 3H). LC (Cond. 1): RT = 1.92 min; LC/MS: Anal. Calcd.
for
[M+H]+ C31H30NO3: 464.45; found 464.52.
The relative stereochemical assignments of the DIBAL-reduction products
were made based on NOE studies conducted on lactone derivatives prepared from
each isomer by employing the following protocol: LiHMDS (50 uL of 1.0 M/THF,
0.05 mmol) was added to a cooled (ice-water) THF (2.0 mL) solution of (2S,3 S)-
benzyl 4-hydroxy-3-methy1-2-(9-pheny1-9H-fluoren-9-ylamino)butanoate (62.7 mg,
0.135 mmol), and the reaction mixture was stirred at similar temperature for
¨2 hr.
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The volatile component was removed in vacuo and the residue was partitioned
between CH2C12 (30 mL), water (20 mL) and saturated aqueous NH4C1 solution (1
mL). The organic layer was dried (MgSO4), filtered, and concentrated in vacuo,
and
the resulting crude material was submitted to a Biotage purification (40 g
silica gel;
10-15% Et0Ac/hexanes) to afford (3S,4S)-4-methy1-3-(9-pheny1-9H-fluoren-9-
ylamino)dihydrofuran-2(31])-one as a colorless film of solid (28.1 mg).
(2S,3R)-
benzyl 4-hydroxy-3-methy1-2-(9-pheny1-9H-fluoren-9-ylamino)butanoate was
elaborated similarly to (3S,4R)-4-methy1-3-(9-pheny1-9H-fluoren-9-
ylamino)dihydrofuran-2(31])-one. (3S,4S)-lactone isomer: 1H NMR (DMSO-d6, 6 =
2.5 ppm, 400 MHz), 7.83 (d, J = 7.5, 2H), 7.46-7.17 (m, 11H), 4.14 (app t, J =
8.3,
1H), 3.60 (d, J = 5.8, NH), 3.45 (app t, J = 9.2, 1H), ¨2.47 (m, 1H, partially
overlapped with solvent signal), 2.16 (m, 1H), 0.27 (d, J= 6.6, 3H). LC (Cond.
1):
RT = 1.98 min; LC/MS: Anal. Calcd. for [M+Na]+ C24H21NNa02: 378.15; found
378.42. (3S,4R)-lactone isomer: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz), 7.89
(d, J = 7.6, 1H), 7.85 (d, J = 7.3, 1H), 7.46-7.20 (m, 11H), 3.95 (dd, J =
9.1, 4.8,
1H), 3.76 (d, J = 8.8, 1H), 2.96 (d, J = 3.0, NH), 2.92 (dd, J = 6.8, 3, NCH),
1.55 (m,
1H), 0.97 (d, J = 7.0, 3H). LC (Cond. 1): RT = 2.03 min; LC/MS: Anal. Calcd.
for
[M+Na]+ C24H21NNa02: 378.15; found 378.49.
TBDMS-Cl (48 mg, 0.312 mmol) followed by imidazole (28.8 mg, 0.423
mmol) were added to a CH2C12 (3 ml) solution of (2S,3S)-benzyl 4-hydroxy-3-
methy1-2-(9-pheny1-9H-fluoren-9-ylamino)butanoate (119.5 mg, 0.258 mmol), and
the mixture was stirred at ambient condition for 14.25 hr. The reaction
mixture was
then diluted with CH2C12 (30 mL) and washed with water (15 mL), and the
organic
layer was dried (MgSO4), filtered, and concentrated in vacuo. The resultant
crude
material was purified with a Biotage (40 g silica gel; 5% Et0Ac/hexanes) to
afford
(2S,3S)-benzyl 4-(tert-butyldimethylsilyloxy)-3-methy1-2-(9-pheny1-9H-fluoren-
9-
ylamino)butanoate, contaminated with TBDMS based impurities, as a colorless
viscous oil (124.4 mg). (2S,3R)-benzyl 4-hydroxy-3-methy1-2-(9-pheny1-9H-
fluoren-9-ylamino)butanoate was elaborated similarly to (2S,3R)-benzyl 4-(tert-
butyldimethylsilyloxy)-3-methy1-2-(9-pheny1-9H-fluoren-9-ylamino)butanoate.
(2S,3S)-sily1 ether isomer: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz), 7.82 (d, J
=
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4.1, 1H), 7.80 (d, J = 4.0, 1H), 7.38-7.07 (m, 16 H), 4.70 (d, J = 12.4, 1H),
4.42 (d, J
= 12.3, 1H), 3.28-3.19 (m, 3H), 2.56 (dd, J = 10.1, 5.5, 1H), 1.61 (m, 1H),
0.90 (d, J
= 6.8, 3H), 0.70 (s, 9H), -0.13 (s, 3H), -0.16 (s, 3H). LC (Cond. 1, where the
run
time was extended to 4 min): RT = 3.26 min; LC/MS: Anal. Calcd. for [M+H]+
C37H44NO3Si: 578.31; found 578.40. (2S,3R)-sily1 ether isomer: 1H NMR (DMSO-
d6, 6 = 2.5 ppm, 400 MHz), 7.82 (d, J = 3.0, 1H), 7.80 (d, J = 3.1, 1H), 7.39-
7.10 (m,
16H), 4.66 (d, J = 12.4, 1H), 4.39 (d, J = 12.4, 1H), 3.61 (dd, J = 9.9, 5.6,
1H), 3.45
(d, J = 9.5, 1H), 3.41 (dd, J = 10, 6.2, 1H), 2.55 (dd, J = 9.5, 7.3, 1H),
1.74 (m, 1H),
0.77 (s, 9H), 0.61 (d, J= 7.1, 3H), -0.06 (s, 3H), -0.08 (s, 3H).
A balloon of hydrogen was attached to a mixture of (2S,3S)-benzyl 4-(tert-
butyldimethylsilyloxy)-3-methy1-2-(9-pheny1-9H-fluoren-9-ylamino)butanoate
(836
mg, 1.447 mmol) and 10% Pd/C (213 mg) in Et0Ac (16 mL) and the mixture was
stirred at room temperature for ¨ 21 hr, where the balloon was recharged with
H2 as
necessary. The reaction mixture was diluted with CH2C12 and filtered through a
pad
30 Water (1 mL) and NaOH (0.18 mL of 1.0 M/H20, 0.18 mmol) were added to
a mixture of (2S,3S)-2-amino-4-(tert-butyldimethylsilyloxy)-3-methylbutanoic
acid
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(41.9 mg, 0.169 mmol) and Na2CO3 (11.9 mg, 0.112 mmol), and sonicated for
about
1 min to effect dissolution of reactants. The mixture was then cooled with an
ice-
water bath, methyl chloroformate (0.02 mL, 0.259 mmol) was added over 30 s,
and
vigorous stirring was continued at similar temperature for 40 min and then at
ambient
temperature for 2.7 hr. The reaction mixture was diluted with water (5 mL),
cooled
with ice-water bath and treated drop-wise with 1.0 N HC1 aqueous solution (-
0.23
mL). The mixture was further diluted with water (10 mL) and extracted with
CH2C12
(15 mL, 2x). The combined organic phase was dried (MgSO4), filtered, and
concentrated in vacuo to afford Cap-80a as an off-white solid. (2S,3R)-2-amino-
4-
(tert-butyldimethylsilyloxy)-3-methylbutanoic acid was similarly elaborated to
Cap-
80b. Cap-80a: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 400 MHz), 12.57 (br s, 1H), 7.64
(d, J = 8.3, 0.3H), 7.19 (d, J = 8.8, 0.7H), 4.44 (dd, J = 8.1, 4.6, 0.3H),
4.23 (dd, J =
8.7, 4.4, 0.7H), 3.56/3.53 (two singlets, 3H), 3.48-3.40 (m, 2H), 2.22-2.10
(m, 1H),
0.85 (s, 9H), ¨0.84 (d, 0.9H, overlapped with t-Bu signal), 0.79 (d, J = 7,
2.1H),
0.02/0.01/0.00 (three overlapping singlets, 6H). LC/MS: Anal. Calcd. for
[M+Na]+
C13H27NNa05Si: 328.16; found 328.46. Cap-80b: 1H NMR (CDC13, 6 = 7.24 PPm,
400 MHz), 6.00 (br d, J = 6.8, 1H), 4.36 (dd, J = 7.1, 3.1, 1H), 3.87 (dd, J =
10.5,
3.0, 1H), 3.67 (s, 3H), 3.58 (dd, J = 10.6, 4.8, 1H), 2.35 (m, 1H), 1.03 (d, J
= 7.1,
3H), 0.90 (s, 9H), 0.08 (s, 6H). LC/MS: Anal. Calcd. for [M+Na]+
C13H22NNa05Si:
328.16; found 328.53. The crude products were utilized without further
purification.
Cap-81
0
H
N
o -"="LOH
0-7
Prepared according to the protocol described by Falb et al. Synthetic
Communications 1993, 23, 2839.
Cap-82 to Cap-85
Cap-82 to Cap-85 were synthesized from appropriate starting materials
according to the procedure described for Cap-51 or Cap-13. The samples
exhibited
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similar spectral profiles as that of their enantiomers (i.e., Cap-4, Cap-13,
Cap-51 and
Cap-52, respectively).
0
H I jj H Cji H Cji
0 N,, N.,... 0 1\14,... 0 1\14,,..
OH )'r OH OH
0 Ph 0 0
Cap-82 Cap-83 Cap-84 Cap-85
Cap-86
Me02CHN
/..._..".1 0
( \OH
OMe
To a mixture of 0-methyl-L-threonine (3.0 g, 22.55 mmol), NaOH (0.902 g,
22.55 mmol) in H20 (15 mL) was added C1CO2Me (1.74 mL, 22.55 mmol) dropwise
at 0 C. The mixture was allowed to stir for 12 h and acidified to pH 1 using
1N HC1.
The aqueous phase was extracted with Et0Ac and (2x250 mL) and 10% Me0H in
CH2C12 (250 mL) and the combined organic phases were concentrated under in
vacuo
to afford a colorless oil (4.18 g, 97%) which was of sufficient purity for use
in
subsequent steps. 1FINMR (400 MHz, CDC13) 6 4.19 (s, 1H), 3.92-3.97 (m, 1H),
3.66 (s, 3H), 1.17 (d, J = 7.7 Hz, 3H). LCMS: Anal. Calcd. for C7H13N05: 191;
found: 190(M-H).
Cap-87
Me02CHN 0
/f
HO OH
To a mixture of L-homoserine (2.0 g, 9.79 mmol), Na2CO3 (2.08 g, 19.59
mmol) in H20 (15 mL) was added C1CO2Me (0.76 mL, 9.79 mmol) dropwise at 0 C.
The mixture was allowed to stir for 48 h and acidified to pH 1 using 1N HC1.
The
aqueous phase was extracted with Et0Ac and (2X250 mL) and the combined organic
phases were concentrated in vacuo to afford a colorless solid (0.719 g, 28%)
which
was of sufficient purity for use in subsequent steps. 1FINMR (400 MHz, CDC13)
6
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4.23 (dd, J= 4.5, 9.1 Hz, 1H), 3.66 (s, 3H), 3.43-3.49 (m, 2H), 2.08 -2.14 (m,
1H),
1.82 - 1.89 (m, 1H). LCMS: Anal. Calcd. for C7H13N05: 191; found: 192 (M+H)+.
Cap-88
--- NH
0
\OH
A mixture of L-valine (1.0 g, 8.54 mmol), 3-bromopyridine (1.8 mL, 18.7
mmol), K2CO3 (2.45 g, 17.7 mmol) and Cut (169 mg, 0.887 mmol) in DMSO (10
mL) was heated at 100 C for 12h. The reaction mixture was cooled to rt, poured
into
H20 (ca. 150 mL) and washed with Et0Ac (x2). The organic layers were extracted
with a small amount of H20 and the combined aq phases were acidified to ca. pH
2
with 6N HC1. The volume was reduced to about one-third and 20g of cation
exchange resin (Strata) was added. The slurry was allowed to stand for 20 min
and
loaded onto a pad of cation exchange resin (Strata) (ca. 25g). The pad was
washed
with H20 (200 mL), Me0H (200 mL), and then NH3 (3M in Me0H, 2X200 mL).
The appropriate fractions was concentrated in vacuo and the residue (ca. 1.1
g) was
dissolved in H20, frozen and lyophyllized. The title compound was obtained as
a
foam (1.02 g, 62%). 11-INMR (400 MHz, DMSO-d6) 6 8.00 (s, br, 1H), 7.68 -7.71
(m, 1H), 7.01 (s, br, 1H), 6.88 (d, J= 7.5 Hz, 1H), 5.75 (s, br, 1H), 3.54 (s,
1H), 2.04
-2.06 (m, 1H), 0.95 (d, J= 6.0 Hz, 3H), 0.91 (d, J= 6.6 Hz, 3H). LCMS: Anal.
Calcd. for C101-114N202: 194; found: 195 (M+H)+.
Cap-89
N
j,õ
0
A mixture of L-valine (1.0 g, 8.54 mmol), 5-bromopyrimidine (4.03 g, 17.0
mmol), K2CO3 (2.40 g, 17.4 mmol) and Cut (179 mg, 0.94 mmol) in DMSO (10 mL)
was heated at 100 C for 12h. The reaction mixture was cooled to RT, poured
into
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H20 (ca. 150 mL) and washed with Et0Ac (x2). The organic layers were extracted
with a small amount of H20 and the combined aq phases were acidified to ca. pH
2
with 6N HC1. The volume was reduced to about one-third and 20g of cation
exchange resin (Strata) was added. The slurry was allowed to stand for 20 min
and
loaded onto a pad of cation exchange resin (Strata) (ca. 25g). The pad was
washed
with H20 (200 mL), Me0H (200 mL), and then NH3 (3M in Me0H, 2x200 mL).
The appropriate fractions was concentrated in vacuo and the residue (ca. 1.1
g) was
dissolved in H20, frozen and lyophyllized. The title compound was obtained as
a
foam (1.02 g, 62%). 1FINMR (400 MHz, CD30D) showed the mixture to contain
valine and the purity could not be estimated. The material was used as is in
subsequent reactions. LCMS: Anal. Calcd. for C9H13N302: 195; found: 196
(M+H)+.
Cap-90
CO2H
0 IlMe2
Cap-90 was prepared according to the method described for the preparation
of Cap-1. The crude material was used as is in subsequent steps. LCMS: Anal.
Calcd. for C11H15NO2: 193; found: 192 (M-H).
The following caps were prepared according to the method used for
preparation of cap Si unless noted otherwise:
Cap Structure LCMS
Cap-91 NHCO2Me LCMS: Anal. Calcd. for
0 CO2H
C11H13N04: 223; found:
222 (M-H).
Cap-92 NHCO2Me LCMS: Anal. Calcd. for
0 - CO2H
C11H13N04: 223; found:
222 (M-H).
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Cap-93 I LCMS: Anal. Calcd. for
0
HNõ C10H12N204: 224; found:
,
).0H 225 (M+H)+.
N
Cap-94 0 LCMS: Anal. Calcd. for
e.)/y(OH CalliN304: 213; found:
N HN 0
H II 214 (M+H)+.
0
Cap-95 0 LCMS: Anal. Calcd. for
0)*LNH 0 C13H17N04: 251; found:
01 OH 250 (M-H)-.
Cap-96 0 LCMS: Anal. Calcd. for
0).1\1H 0 C12H15N04: 237; found:
OH 236 (M-H).
101
Cap-97 0 LCMS: Anal. Calcd. for
0),I\IH0 C9H15N04: 201; found:
a'AOH 200 (M-H).
Cap-98 0 LCMS: Anal. Calcd. for
OANH 0 C9H15N04: 201; found:
arLLOH 202 (M+H)+.
Cap-99 0 11-1NMR (400 MHz,
0)L NH CD30D) 6 3.88 - 3.94 (m,
CO2 1H), 3.60, 3.61 (s, 3H),
H
2.80 (m, 1H), 2.20 (m 1H),
1.82- 1.94 (m, 3H), 1.45
- 1.71 (m, 2H).
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Cap-99a 0 11-11\IMR (400 MHz,
OA NH CD30D) 6 3.88 ¨ 3.94 (m,
0 1H), 3.60, 3.61 (s, 3H),
tO2H 2.80 (m, 1H), 2.20 (m 1H),
1.82¨ 1.94 (m, 3H), 1.45
¨ 1.71 (m, 2H).
Cap-100 0 LCMS: Anal. Calcd. for
OANH 0 C12H14N04F: 255; found:
OH 256 (M+H)+.
F el
Cap-101 0 LCMS: Anal. Calcd. for
0).LNH C11H13N04: 223; found:
CO2H 222 (M-H)-.
101
Cap-102 0 LCMS: Anal. Calcd. for
0).LNH C11H13N04: 223; found:
CO2H 222 (M-H)-
101
Cap-103 0 LCMS: Anal. Calcd. for
0)-NH C10H12N204: 224; found:
CO2H 225 (M+H)+.
N
Cap-104 HNN-0-.CO2H 11-11\IMR (400 MHz,
0 CD30D) 6 3.60 (s, 3H),
0
/ 3.50 ¨ 3.53 (m, 1H), 2.66
¨2.69 and 2.44 ¨ 2.49 (m,
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1H), 1.91 -2.01 (m, 2H),
1.62- 1.74 (m, 4H), 1.51
- 1.62 (m, 2H).
Cap-105 11-1NMR (400 MHz,
HNII,Ø1CO2H
0 CD30D) 6 3.60 (s, 3H),
0
/ 3.33 - 3.35 (m, 1H,
partially obscured by
solvent), 2.37 - 2.41 and
2.16 - 2.23 (m, 1H), 1.94
-2.01 (m, 4H), 1.43 -
1.53 (m, 2H), 1.17- 1.29
(m, 2H).
Cap-106 -\N...Ø....
CO2H 11-1NMR (400 MHz,
CD30D) 6 3.16 (q, J =
7.3 Hz, 4H), 2.38 - 2.41
Prepared from cis-4-
(m, 1H), 2.28 - 2.31 (m,
aminocyclohexane 2H), 1.79 - 1.89 (m, 2H),
carboxylic acid and 1.74 (app, ddd J = 3.5,
acetaldehyde by 12.5, 15.9 Hz, 2H), 1.46
employing a similar
(app dt J = 4.0, 12.9 Hz,
procedure described for 2H), 1.26 (t, J = 7.3 Hz,
the synthesis of Cap-2.
6H)
The crude HC1 salt was
passed through MCX
(Me0H/H20/CH2C12
wash; 2 N NH3/Me0H
elution) to afford an oil,
which was dissolved in
CH3CN/H20 and
lyophilized to afford a
tan solid.
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Cap-107 0 LCMS: Anal. Calcd. for
e.n)(OH C8H10N204S: 230; found:
s HNO
II 231 (M+H)+.
0
Cap-108 0 LCMS: Anal. Calcd. for
e_yY(OH C15H17N304: 303; found:
N HN 0
y 304 (M+H)+.
Ph-1 0
Cap-109 0 LCMS: Anal. Calcd. for
0)-NH C10H12N204: 224; found:
CO2H 225 (M+H)+.
IN
Cap-110 0 LCMS: Anal. Calcd. for
0)=NH C10H12N204: 224; found:
CO2H 225 (M+H)+.
I
N
Cap-ill 0 LCMS: Anal. Calcd. for
0)LNH C12H16N08P: 333; found:
CO2H 334 (M+H)+.
Rµ ,0
P
Me0- \OH
Cap-112 0 LCMS: Anal. Calcd. for
0)LNH C13H14N204: 262; found:
CO2H 263 (M+H)+.
4110 NNH
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Cap-113 0 LCMS: Anal. Calcd. for
0)-NH C18H19N05: 329; found:
CO2H 330 (M+H)+.
101
OBn
Cap-114 ,002Me 1141\IMR (400 MHz,
i¨N1
' CDC13) 6 4.82 ¨ 4.84 (m,
CO2H
1H), 4.00 ¨ 4.05 (m, 2H),
3.77 (s, 3H), 2.56 (s, br,
2H)
4'=(CO2H
Cap-115 1141\IMR (400 MHz,
NHCO2Me CDC13) 6 5.13 (s, br, 1H),
4.13 (s, br, 1H), 3.69 (s,
3H),2.61 (d, J = 5.0 Hz,
2H), 1.28 (d, J = 9.1 Hz,
3H).
Cap-116 1141\IMR (400 MHz,
/4`=-rCO2H CDC13) 6 5.10 (d, J= 8.6
NHCO2Me
Hz, 1H), 3.74 ¨ 3.83 (m,
1H), 3.69 (s, 3H), 2.54 ¨
2.61 (m, 2H), 1.88 (sept, J
= 7.0 Hz, 1H), 0.95 (d, J =
7.0 Hz, 6H).
Cap-117 to Cap-123
For the preparation of Cap-117 to Cap-123 the Boc amino acids were
obtained from commercially sources and were deprotected by treatment with 25%
TFA in CH2C12. After complete reaction as judged by LCMS the solvents were
removed in vacuo and the corresponding TFA salt of the amino acid was
carbamoylated with methyl chloroformate according to the procedure described
for
Cap-51.
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Cap Structure LCMS
Cap-117 0 LCMS: Anal.
Calcd.
0)"LNH 0 for
C12H15N04: 237;
OH found: 238 (M+H)+.
I.
Cap-118 0 LCMS: Anal.
Calcd.
0).LI\IH 0 for
C10H13N04S: 243;
C)OH found: 244 (M+H)+.
(IS
Cap-119 0 LCMS: Anal.
Calcd.
0).LI\IH 0 for
C10H13N04S: 243;
OH found: 244 (M+H)+.
(IS
Cap-120 0 LCMS: Anal.
Calcd.
0)*(NH 0 for
C10H13N04S: 243;
C)OH found: 244 (M+H)+.
0 \ S
Cap-121 0 11-1NMR (400
MHz,
0)-LNH CDC13) 6 4.06
¨4.16
n.....0O2H (m, 1H), 3.63 (s, 3H),
3.43 (s, 1H), 2.82 and
2.66 (s, br, 1H), 1.86 ¨
2.10 (m, 3H), 1.64 ¨
1.76 (m, 2H), 1.44 ¨
1.53 (m, 1H).
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Cap-122 0 11-1NMR profile is
0)LNH similar to that of its
enantiomer, Cap-121.
Cap-123 o,
o/ LCMS: Anal. Calcd.
>1¨NH
0 41 for C27H26N206: 474;
4.46 0 ..INH
found: 475 (M+H)+.
latir OH
Cap-124
0
(
_i___=
0 ' OH
0
The hydrochloride salt of L-threonine tert-butyl ester was carbamoylated
according to the procedure for Cap-51. The crude reaction mixture was
acidified
with 1N HC1 to pH-1 and the mixture was extracted with Et0Ac (2X50 mL). The
combined organic phases were concentrated in vacuo to give a colorless oil
which
solidified on standing. The aqueous layer was concentrated in vacuo and the
resulting mixture of product and inorganic salts was triturated with Et0Ac-
CH2C12-
Me0H (1:1:0.1) and then the organic phase concentrated in vacuo to give a
colorless
oil which was shown by LCMS to be the desired product. Both crops were
combined
to give 0.52 g of a solid. 11-1NMR (400 MHz, CD30D) 6 4.60 (m, 1H), 4.04 (d, J
=
5.0 Hz, 1H), 1.49 (d, J = 6.3 Hz, 3H). LCMS: Anal. Calcd. for C5H7N04: 145;
found: 146 (M+H)+.
Cap-125
/
--N 0
\----)-1(OH
BocHN
To a suspension of Pd(OH)2, (20%, 100 mg), aqueous formaldehyde (37% wt,
4 ml), acetic acid, (0.5 mL) in methanol (15 mL) was added (S)-4-amino-2-(tert-
butoxycarbonylamino)butanoic acid (1 g, 4.48 mmol). The reaction was purged
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several times with hydrogen and was stirred overnight with an hydrogen balloon
room temp. The reaction mixture was filtered through a pad of diatomaceous
earth
(Celite()), and the volatile component was removed in vacuo. The resulting
crude
material was used as is for the next step. LC/MS: Anal. Calcd. for C11H22N204:
246; found: 247 (M+H)+.
Cap-126
4¨NM e /r-NMe
CICO2Me, NaHCO3 Nv_j1
THF / H20 / 0 C Me02CHN CO2H
H2N CO2H
cj-25 cap-126
This procedure is a modification of that used to prepare Cap-51. To a
suspension of 3-methyl-L-histidine (0.80 g, 4.70 mmol) in THF (10mL) and H20
(10
mL) at 0 C was added NaHCO3 (0.88 g, 10.5 mmol). The resulting mixture was
treated with C1CO2Me (0.40 mL, 5.20 mmol) and the mixture allowed to stir at 0
C.
After stirring for ca. 2h LCMS showed no starting material remaining. The
reaction
was acidified to pH 2 with 6 N HC1.
The solvents were removed in vacuo and the residue was suspended in 20 mL
of 20% Me0H in CH2C12. The mixture was filtered and concentrated to give a
light
yellow foam (1.21 g,). LCMS and 1H NMR showed the material to be a 9:1 mixture
of the methyl ester and the desired product. This material was taken up in THF
(10mL) and H20 (10mL), cooled to 0 C and LiOH (249.1 mg, 10.4 mmol) was
added. After stirring ca. lh LCMS showed no ester remaining. Therefore the
mixture was acidified with 6N HC1 and the solvents removed in vacuo. LCMS and
1H NMR confirm the absence of the ester. The title compound was obtained as
its
HC1 salt contaminated with inorganic salts (1.91 g, >100%). The compound was
used as is in subsequent steps without further purification. 1HNMR (400 MHz,
CD30D) 6 8.84, (s, 1H), 7.35 (s, 1H), 4.52 (dd, J = 5.0, 9.1 Hz, 1H), 3.89 (s,
3H),
3.62 (s, 3H), 3.35 (dd, J = 4.5, 15.6 Hz, 1H, partially obscured by solvent),
3.12 (dd,
J = 9.0, 15.6 Hz, 1H).LCMS: Anal. Calcd. for C9H13N304: 227.09; found: 228.09
(M+H)+.
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Cap-127
MN MeN
CICO2Me, NaHCO3
THF / H20 / 0 C Me02CHN CO2H
H2N CO2H
cj-26 cap-127
Cap-127 was prepared according to the method for Cap-126 above starting
from (S)-2-amino-3-(1-methyl-1H-imidazol-4-y1)propanoic acid (1.11 g, 6.56
mmol),
NaHCO3 (1.21 g, 14.4 mmol) and C1CO2Me (0.56 mL, 7.28 mmol). The title
compound was obtained as its HC1 salt (1.79 g, >100%) contaminated with
inorganic
salts. LCMS and 1H NMR showed the presence of ca. 5% of the methyl ester. The
crude mixture was used as is without further purification. 1HNMR (400 MHz,
CD30D) 6 8.90 (s, 1H), 7.35 (s, 1H), 4.48 (dd, J = 5.0, 8.6 Hz, 1H), 3.89 (s,
3H),
3.62 (s, 3H), 3.35 (m, 1H), 3.08 (m, 1H); LCMS: Anal. Calcd. for C9H13N304:
227.09; found: 228 (M+H)+.
Preparation of Cap-128
Ph¨\
Cbz-CI / DMAP
BnBr / CuSO4-5H20
CH2Cl2 / iPr2NEt Nis
0 C sodium ascorbate
BocHNCO2H BocHN CO2Bn
NaN3 / DMF / H20 BocHN CO2Bn
cj-27a cj-27b 65 C / 12 h cj-28
Ph¨\
HN
1) TFA / CH2Cl2 N. H2 / Pd-C
N.
Me0H
2) CICO2Me / NaHCO3
THF-H20 Me02CHN CO2Bn Me02CHN
CO2H
cj-29 cap-128
Step 1. Preparation of (S)-benzyl 2-(tert-butoxycarbonylamino)pent-4-ynoate
(cj-
27b).
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BocH N LCO2Bn
cj-27b
To a solution of cj-27a (1.01 g, 4.74 mmol), DMAP (58 mg, 0.475 mmol) and
iPr2NEt (1.7 mL, 9.8 mmol) in CH2C12 (100 mL) at 0 C was added Cbz-C1 (0.68
mL,
4.83 mmol). The solution was allowed to stir for 4 h at 0 C, washed (1N KHSO4,
brine), dried (Na2SO4), filtered, and concentrated in vacuo. The residue was
purified
by flash column chromatography (TLC 6:1 hex:Et0Ac) to give the title compound
(1.30 g, 91%) as a colorless oil. 1FINMR (400 MHz, CDC13) 6 7.35 (s, 5H), 5.35
(d,
br, J = 8.1 Hz, 1H), 5.23 (d, J = 12.2 Hz, 1H), 5.17 (d, J = 12.2 Hz, 1H),
4.48 - 4.53
(m, 1H), 2.68 -2.81 (m, 2H), 2.00 (t, J= 2.5 Hz, 1H), 1.44 (s, 9H). LCMS:
Anal.
Step 2. Preparation of (S)-benzyl 3-(1-benzy1-1H-1,2,3-triazol-4-y1)-2-(tert-
butoxycarbonylamino)propanoate (cj-28).
Ph -
N.
31
sN
BocH N CO2Bn
cj-28
To a mixture of (S)-benzyl 2-(tert-butoxycarbonylamino)pent-4-ynoate (0.50
g, 1.65 mmol), sodium ascorbate (0.036 g, 0.18 mmol), CuSO4-5H20 (0.022 g,
0.09
mmol) and NaN3 (0.13 g, 2.1 mmol) in DMF-H20 (5 mL, 4:1) at rt was added BnBr
(0.24 mL, 2.02 mmol) and the mixture was warmed to 65 C. After 5h LCMS
indicated low conversion. A further portion of NaN3 (100 mg) was added and
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suggests the presence of DMF. The material was used as is without further
purification. 1HNMR (400 MHz, DMSO-d6) 6 7.84 (s, 1H), 7.27 - 7.32 (m, 10H),
5.54 (s, 2H), 5.07 (s, 2H), 4.25 (m, 1H), 3.16 (dd, J = 1.0, 5.3 Hz, 1H), 3.06
(dd, J =
5.3, 14.7 Hz), 2.96 (dd, J = 9.1, 14.7 Hz, 1H), 1.31 (s, 9H).
LCMS: Anal. Calcd. for C24H28N404: 436; found: 437 (M+H)+.
Step 3. Preparation of (S)-benzyl 3-(1-benzy1-1H-1,2,3-triazol-4-y1)-2-
(methoxycarbonylamino)propanoate (cj-29).
Ph-\
NI, 31
µ1\1
Me02CHN CO2Bn
cj-29
A solution of (S)-benzyl 3-(1-benzy1-1H-1,2,3-triazol-4-y1)-2-(tert-
butoxycarbonylamino)propanoate (0.52 g, 1.15 mmol) in CH2C12 was added TFA (4
mL). The mixture was allowed to stir at room temperature for 2h. The mixture
was
concentrated in vacuo to give a colorless oil which solidified on standing.
This
material was dissolved in THF-H20 and cooled to 0 C. Solid NaHCO3 (0.25 g,
3.00
mmol) was added followed by C1CO2Me (0.25 mL, 3.25 mmol). After stirring for
1.5h the mixture was acidified to pH-2 with 6N HC1 and then poured into H20-
Et0Ac. The layers were separated and the aq phase extracted 2x with Et0Ac. The
combined org layers were washed (H20, brine), dried (Na2504), filtered, and
concentrated in vacuo to give a colorless oil (505.8 mg, 111%, NMR suggested
the
presence of an unidentified impurity) which solidified while standing on the
pump.
The material was used as is without further purification. 1FINMR (400 MHz,
DMSO-
d6) 6 7.87 (s, 1H), 7.70 (d, J = 8.1 Hz, 1H), 7.27 - 7.32 (m, 10H), 5.54 (s,
2H), 5.10
(d, J = 12.7 Hz, 1H), 5.06 (d, J = 12.7 Hz, 1H), 4.32 -4.37 (m, 1H), 3.49 (s,
3H),
3.09 (dd, J= 5.6, 14.7 Hz, 1H), 2.98 (dd, J= 9.6, 14.7 Hz, 1H). LCMS: Anal.
Calcd. for C21H22N404: 394; found: 395 (M+H)+.
Step 4. Preparation of (S)-2-(methoxycarbonylamino)-3-(1H-1,2,3-triazol-4-
yl)propanoic acid (Cap-128).
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HN
N.
Me02CHN CO2H
Cap-128
(S)-benzyl 3-(1-benzy1-1H-1,2,3-triazol-4-y1)-2-
(methoxycarbonylamino)propanoate (502 mg, 1.11 mmol) was hydrogenated in the
presence of Pd-C (82 mg) in Me0H (5 mL) at atmospheric pressure for 12h. The
mixture was filtered through diatomaceous earth (Celite ) and concentrated in
vacuo.
(S)-2-(methoxycarbonylamino)-3-(1H-1,2,3-triazol-4-yl)propanoic acid was
obtained
as a colorless gum (266 mg, 111%) which was contaminated with ca. 10% of the
methyl ester. The material was used as is without further purification. 1FINMR
(400
MHz, DMSO-d6) 6 12.78 (s, br, 1H), 7.59 (s, 1H), 7.50 (d, J= 8.0 Hz, 1H), 4.19
¨
4.24 (m, 1H), 3.49 (s, 3H), 3.12 (dd, J = 4.8 Hz, 14.9 Hz, 1H), 2.96 (dd, J =
9.9, 15.0
Hz, 1H). LCMS: Anal. Calcd. for C7H10N404: 214; found: 215 (M+H)+.
Preparation of Cap-129
N / 1) H2 Pd-C / Me0H
CbzHN 0 CH3CN / 50 C CbzHN CO2H 2) CICO2Me
MeO2CHNCO2H
X
cj-30 NaHCO3 / THF-H20
cj-31 cap-129
Step 1. Preparation of (S)-2-(benzyloxycarbonylamino)-3-(1H-pyrazol-1-
yl)propanoic acid (cj-31).
Ni
CbzHNCO2H
cj-31
A suspension of (S)-benzyl 2-oxooxetan-3-ylcarbamate (0.67 g, 3.03 mmol),
and pyrazole (0.22 g, 3.29 mmol) in CH3CN (12 mL) was heated at 50 C for 24h.
The mixture was cooled to rt overnight and the solid filtered to afford (S)-2-
(benzyloxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid (330.1 mg). The
filtrate was concentrated in vacuo and then triturated with a small amount of
CH3CN
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(ca. 4 mL) to afford a second crop (43.5 mg). Total yield 370.4 mg (44%). m.p.
165.5 - 168 C. lit m.p. 168.5 - 169.5 [Vederas et al. J. Am. Chem. Soc. 1985,
107, 7105]. 11-INMR (400 MHz, CD30D) 6 7.51 (d, J = 2.0, 1H), 7.48 (s, J = 1.5
Hz,
1H), 7.24 - 7.34 (m, 5H), 6.23 m, 1H), 5.05 (d, 12.7 H, 1H), 5.03 (d, J = 12.7
Hz,
1H), 4.59 - 4.66 (m, 2H), 4.42 - 4.49 (m, 1H). LCMS: Anal. Calcd. for
C14H15N304: 289; found: 290 (M+H)+.
Step 2. Preparation of (S)-2-(methoxycarbonylamino)-3-(1H-pyrazol-1-
yl)propanoic
acid (Cap-129).
11
N /
r
MeO2CHNCO2H
cap-129
(S)-2-(benzyloxycarbonylamino)-3-(1H-pyrazol-1-yl)propanoic acid (0.20 g,
0.70 mmol) was hydrogenated in the presence of Pd-C (45 mg) in Me0H (5 mL) at
atmospheric pressure for 2h. The product appeared to be insoluble in Me0H,
therefore the reaction mixture was diluted with 5mL H20 and a few drops of 6N
HC1.
The homogeneous solution was filtered through diatomaceous earth (Celite(D),
and
the Me0H removed in vacuo. The remaining solution was frozen and lyophyllized
to
give a yellow foam (188.9 mg). This material was suspended in THF-H20 (1:1,
10mL) and then cooled to 0 C. To the cold mixture was added NaHCO3 (146.0 mg,
1.74 mmol) carefully (evolution of CO2). After gas evolution had ceased (ca.
15
min) C1CO2Me (0.06 mL, 0.78 mmol) was added dropwise. The mixture was
allowed to stir for 2h and was acidified to pH-2 with 6N HC1 and poured into
Et0Ac. The layers were separated and the aqueous phase extracted with EtOAC
(x5). The combined organic layers were washed (brine), dried (Na2SO4),
filtered,
and concentrated to give the title compound as a colorless solid (117.8 mg,
79%).
11-INMR (400 MHz, DMSO-d6) 6 13.04 (s, 1H), 7.63 (d, J = 2.6 Hz, 1H), 7.48 (d,
J =
8.1 Hz, 1H), 7.44 (d, J = 1.5 Hz, 1H), 6.19 (app t, J = 2.0 Hz, 1H), 4.47 (dd,
J = 3.0,
12.9 Hz, 1H), 4.29 -4.41 (m, 2H), 3.48 (s, 3H). LCMS: Anal. Calcd. for
C8R1N304: 213; found: 214 (M+H)+.
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Cap-130
401
AcHNCO2H
Cap-130 was prepared by acylation of commercially available (R)-
phenylglycine analgous to the procedure given in: Calmes, M.; Daunis, J.;
Jacquier,
R.; Verducci, J. Tetrahedron, 1987, 43(10), 2285.
Cap-131
O
0 0,Bn H
0
¨.-
b0
HNy0
NH2 HNy0
HOI N N
..., --..
..-- =-..
Step a: Dimethylcarbamoyl chloride (0.92 mL, 10 mmol) was added slowly to
a solution of (S)-benzyl 2-amino-3-methylbutanoate hydrochloride (2.44 g; 10
mmol)
and Hunig's base (3.67 mL, 21 mmol) in THF (50 mL). The resulting white
suspension was stirred at room temperature overnight (16 hours) and
concentrated
under reduced pressure. The residue was partitioned between ethyl acetate and
water. The organic layer was washed with brine, dried (MgSO4), filtered, and
concentrated under reduced pressure. The resulting yellow oil was purified by
flash
chromatography, eluting with ethyl acetate:hexanes (1:1). Collected fractions
were
concentrated under vacuum providing 2.35 g (85%) of clear oil. 1H NMR (300
MHz,
DMSO-d6) 6 ppm 0.84 (d, J=6.95 Hz, 3H), 0.89 (d, J=6.59 Hz, 3H), 1.98-2.15 (m,
1H), 2.80 (s, 6H), 5.01-5.09 (m, J=12.44 Hz, 1H), 5.13 (d, J=12.44 Hz, 1H),
6.22 (d,
J=8.05 Hz, 1H), 7.26-7.42 (m, 5H). LC (Cond. 1): RT = 1.76 min; MS: Anal.
Calcd. for [M+H]+ C16H22N203: 279.17; found 279.03.
Step b: To a Me0H (50 mL) solution of the intermediate prepared above
(2.35 g; 8.45 mmol) was added Pd/C (10%; 200 mg) and the resulting black
suspension was flushed with N2 (3x) and placed under 1 atm of H2. The mixture
was
stirred at room temperature overnight and filtered though a microfiber filter
to
remove the catalyst. The resulting clear solution was then concentrated under
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reduced pressure to obtain 1.43 g (89%) of Cap-131 as a white foam, which was
used
without further purification. 1H NMR (500 MHz, DMSO-d6) 6 ppm 0.87 (d, J=4.27
Hz, 3H), 0.88 (d, J=3.97 Hz, 3H), 1.93-2.11 (m, 1H), 2.80 (s, 6H), 3.90 (dd,
J=8.39,
6.87 Hz, 1H), 5.93 (d, J=8.54 Hz, 1H), 12.36 (s, 1H). LC (Cond. 1): RT = 0.33
min;
MS: Anal. Calcd. for [M+H]+ C8H17N203: 189.12; found 189.04.
Cap-132
OH
0
YLO
.-
yLo,Bn -
HN y0
NH2
HCI 1\k
Cap-132 was prepared from (S)-benzyl 2-aminopropanoate hydrochloride
according to the method described for Cap-131. 1H NMR (500 MHz, DMSO-d6) 6
ppm 1.27 (d, J=7.32 Hz, 3H), 2.80 (s, 6H), 4.06 (qt, 1H), 6.36 (d, J=7.32 Hz,
1H),
12.27 (s, 1H). LC (Cond. 1): RT = 0.15 min; MS: Anal. Calcd. for [M+H]+
C6H13N203: 161.09; found 161.00.
Cap-133
OH
0
0
NH2 HNy0
HCI 0.õ,...=\ F
Cap-133 was prepared from (S)-tert-butyl 2-amino-3-methylbutanoate
hydrochloride and 2-fluoroethyl chloroformate according to the method
described for
Cap-47. 1H NMR (500 MHz, DMSO-d6) 6 ppm 0.87 (t, J = 6.71 Hz, 6H), 1.97-2.10
(m, 1H), 3.83 (dd, J=8.39, 5.95 Hz, 1H), 4.14-4.18 (m, 1H), 4.20-4.25 (m, 1H),
4.50-
4.54 (m, 1H), 4.59-4.65 (m, 1H), 7.51 (d, J = 8.54 Hz, 1H), 12.54 (s, 1H).
Cap-134
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0 OH
OH-,.- .....................
NH2 HN 0
r
0
Cap-134 was prepared from (S)-diethyl alanine and methyl chloroformate
according to the method described for Cap-51. 1H NMR (500 MHz, DMSO-d6) 6
ppm 0.72-0.89 (m, 6H), 1.15-1.38 (m, 4H), 1.54-1.66 (m, 1H), 3.46-3.63 (m,
3H),
4.09 (dd, J= 8.85, 5.19 Hz, 1H), 7.24 (d, J= 8.85 Hz, 1H), 12.55 (s, 1H). LC
(Cond. 2): RT = 0.66 min; LC/MS: Anal. Calcd. for [M+H]+ C9H18N04: 204.12;
found 204.02.
Cap-135
0
el F
HO
N
A solution of D-2-amino-(4-fluorophenyl)acetic acid (338 mg, 2.00 mmol),
1N HC1 in diethylether (2.0 mL, 2.0 mmol) and formalin (37%, 1 mL) in methanol
(5
mL) was subjected to balloon hydrogenation over 10% palladium on carbon (60
mg)
for 16 h at 25 C. The mixture was then filtered through Celite to afford the
HC1 salt
of Cap-135 as a white foam (316 mg, 80%). 1H NMR (300 MHz, Me0H-d4) 6 7.59
(dd, J= 8.80, 5.10 Hz, 2H), 7.29 (t, J= 8.6 Hz, 2H), 5.17 (s, 1H), 3.05 (v br
s, 3H),
2.63 (v br s, 3H); Rt = 0.19 min (Cond.-MS-W5); 95% homogenity index; LRMS:
Anal. Calcd. for [M+H]+ C10H13FN02: 198.09; found: 198.10.
Cap-136
N 0
1N'40H
*
To a cooled (-50 C) suspension of 1-benzy1-1H-imidazole (1.58 g, 10.0
mmol) in anhydrous diethyl ether (50 mL) under nitrogen was added n-butyl
lithium
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(2.5 M in hexanes, 4.0 mL, 10.0 mmol) dropwise. After being stirred for 20 min
at -
50 C, dry carbon dioxide (passed through Drierite) was bubbled into the
reaction
mixture for 10 min before it was allowed to warm up to 25 C. The heavy
precipitate
which formed on addition of carbon dioxide to the reaction mixture was
filtered to
yield a hygroscopic, white solid which was taken up in water (7 mL), acidified
to pH
= 3, cooled, and induced to crystallize with scratching. Filtration of this
precipitate
gave a white solid which was suspended in methanol, treated with 1N
HC1/diethyl
ether (4 mL) and concentrated in vacuo. Lyophilization of the residue from
water (5
mL) afforded the HC1 salt of Cap-136 as a white solid (817 mg, 40%). 1H NMR
(300
MHz, DMSO-d6) 6 7.94 (d, J= 1.5 Hz, 1H), 7.71 (d, J= 1.5 Hz, 1H), 7.50-7.31
(m,
5H), 5.77 (s, 2H); Rt = 0.51 min (Cond.-MS-W5); 95% homogenity index; LRMS:
Anal. Calc. for [M+H]+ C11H12N202: 203.08; found: 203.11.
Cap-137
elN CN
co2H
Cap-137, step a
lelN CN
'0
_
A suspension of 1-chloro-3-cyanoisoquinoline (188 mg, 1.00 mmol; prepared
according to the procedure in WO 2003/ 099274) (188 mg, 1.00 mmol), cesium
fluoride (303.8 mg, 2.00 mmol), bis(tri-tert-butylphosphine)palladium
dichloride (10
mg, 0.02 mmol) and 2-(tributylstannyl)furan (378 L, 1.20 mmol) in anhydrous
dioxane (10 mL) under nitrogen was heated at 80 C for 16 h before it was
cooled to
C and treated with saturated, aqueous potassium fluoride solution with
vigorous
25 stirring for 1 h. The mixture was partitioned between ethyl acetate and
water and the
organic phase was separated, washed with brine, dried over Na2SO4, filtered
and
concentrated. Purification of the residue on silica gel (elution with 0% to
30% ethyl
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acetate/hexanes) afforded Cap-137, step a (230 mg, 105%) as a white solid
which
was carried forward directly. Rt = 1.95 min (Cond.-MS-W2); 90% homogeneity
index; LRMS: Anal. Calc. for [M+I-1]+ C14F181\120: 221.07; found: 221.12.
Cap-137
To a suspension of Cap 137, step a, (110 mg, 0.50 mmol) and sodium
periodate (438 mg, 2.05 mmol) in carbon tetrachloride (1 mL), acetonitrile (1
mL)
and water (1.5 mL) was added ruthenium trichloride hydrate (2 mg, 0.011 mmol).
The mixture was stirred at 25 C for 2 h and then partitioned between
dichloromethane and water. The aqueous layer was separated, extracted twice
more
with dichloromethane and the combined dichloromethane extracts were dried over
Na2SO4, filtered and concentrated. Trituration of the residue with hexanes
afforded
Cap-137 (55 mg, 55%) as a grayish-colored solid. Rt = 1.10 min (Cond.-MS-W2);
90% homogeneity index; LCMS: Anal. Calc. for [M+I-1]+ C11F18N202: 200.08;
found: 200.08.
Caps 138 to 158
Synthetic Strategy. Method A.
OH 0 0 0 0
40 ' DEAD
..... N M eHOF H 0 -,.. mCPBA 0 .õ.. TMSCN 0 -,.... 5N NaOH
T
,,,N DCM\
...., N.: _ TEA, ACN ,, N 85 C 40 -N
BMCL 2001,
11, 1885-1888 CN CO2N
Cap-138
'o
0
N
CO2H
Cap-138, step a
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o
0
To a stirred suspension of 5-hydroxyisoquinoline (prepared according to the
procedure in WO 2003/ 099274) (2.0 g, 13.8 mmol) and triphenylphosphine (4.3
g,
16.5 mmol) in dry tetrahydrofuran (20 mL) was added dry methanol (0.8 mL) and
diethyl azodicarboxylate (3.0 mL, 16.5 mmol) portionwise. The mixture was
stirred
at room temperature for 20 h before it was diluted with ethyl acetate and
washed with
brine, dried over Na2SO4, filtered and concentrated. The residue was
preabsorbed
onto silica gel and chromatographed (elution with 40% ethyl acetate/hexanes)
to
afford Cap-138, step a (1.00 g, 45%) as a light yellow solid. 1H NMR (CDC13,
500
MHz) 6 9.19 (s, 1H), 8.51 (d, J= 6.0 Hz, 1H), 7.99 (d, J= 6.0 Hz, 1H), 7.52-
7.50 (m,
2H), 7.00-6.99 (m, 1H), 4.01 (s, 3H); Rt= 0.66 min (Cond.-D2); 95% homogeneity
index; LCMS: Anal. Calc. for [M+1-1]+ CloHioNO: 160.08; found 160.1.
Cap-138, step b
_
el 1`1+
`o
To a stirred solution of Cap 138, step a (2.34 g, 14.7 mmol) in anhydrous
dichloromethane (50 mL) at room temperature was added meta-chloroperbenzoic
acid (77%, 3.42 g, 19.8 mmol) in one portion. After being stirred for 20 h,
powdered
potassium carbonate (2.0 g) was added and the mixture was stirred for 1 h at
room
temperature before it was filtered and concentrated in vacuo to afford Cap-
138, step
b (2.15 g, 83%) as a pale, yellow solid which was sufficiently pure to carry
forward
directly. 1H NMR (CDC13, 400 MHz) 6 8.73 (d, J= 1.5 Hz, 1H), 8.11 (dd, J= 7.3,
1.7 Hz, 1H), 8.04 (d, J=7.1 Hz, 1H), 7.52 (t, J= 8.1 Hz, 1H), 7.28 (d, J = 8.3
Hz,
1H), 6.91 (d, J= 7.8 Hz, 1H), 4.00 (s, 3H); Rt= 0.92 min, (Cond.-D1); 90%
homogenity index; LCMS: Anal. Calc. for [M+H]P C10H10NO2: 176.07; found:
176Ø
Cap-138, step c
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o
0
CN
To a stirred solution of Cap 138, step b (0.70 g, 4.00 mmol) and triethylamine
(1.1 mL, 8.00 mmol) in dry acetonitrile (20 mL) at room temperature under
nitrogen
was added trimethylsilylcyanide (1.60 mL, 12.00 mmol). The mixture was heated
at
75 C for 20 h before it was cooled to room temperature, diluted with ethyl
acetate
and washed with saturated sodium bicarbonate solution and brine prior to
drying over
Na2SO4 and solvent concentration. The residue was flash chromatographed on
silica
gel (gradient elution with 5% ethyl acetate in hexanes to 25% ethyl acetate in
hexanes) to afford Cap-138, step c (498.7 mg, 68%) as a white, crystalline
solid
along with 223 mg (30%) of additional Cap-138, step c recovered from the
filtrate.
1H NMR (CDC13, 500 MHz) 6 8.63 (d, J= 5.5 Hz, 1H), 8.26 (d, J= 5.5 Hz, 1H),
7.88 (d, J= 8.5 Hz, 1H), 7.69 (t, J= 8.0 Hz, 1H), 7.08 (d, J= 7.5 Hz, 1H),
4.04 (s,
3H); Rt= 1.75 min, (Cond.-D1); 90% homogeneity index; LCMS: Anal. Calc. for
[M+H]+ C11H9N20: 185.07; found: 185.10.
Cap-138
Cap-138, step c (0.45 g, 2.44 mmol) was treated with 5N sodium hydroxide
solution (10 mL) and the resulting suspension was heated at 85 C for 4 h,
cooled to
C, diluted with dichloromethane and acidified with 1N hydrochloric acid. The
20 organic phase was separated, washed with brine, dried over Na2SO4,
concentrated to
1/4 volume and filtered to afford Cap-138 (0.44g, 88.9%) as a yellow solid. 1H
NMR
(DMSO-d6, 400 MHz) 6 13.6 (br s, 1H), 8.56 (d, J= 6.0 Hz, 1H), 8.16 (d, J =
6.0
Hz, 1H), 8.06 (d, J= 8.8 Hz, 1H), 7.71-7.67 (m, 1H), 7.30 (d, J= 8.0 Hz, 1H),
4.02
(s, 3H); Rt = 0.70 min (Cond.-D1); 95% homogenity index; LCMS: Anal. Calc. for
25 [M+H]+ C11H10NO3: 204.07; found: 204.05.
Synthetic Strategy. Method B (derived from Tetrahedron Letters, 2001, 42,
6707).
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oI I oI
0 ______________________________ so ,.... 5N NaOH so ,
,... N Pd(0A02 .....- N
TMEDA, 150 C
CI toluene ON 002H
WO 2003/ 099274
Cap-139
o
0
N
CO2H
Cap-139, step a
o
soN
ON
To a thick-walled, screw-top vial containing an argon-degassed suspension
of 1-chloro-6-methoxyisoquinoline (1.2 g, 6.2 mmol; prepared according to the
procedure in WO 2003/ 099274), potassium cyanide (0.40 g, 6.2 mmol), 1,5-
bis(diphenylphosphino)pentane (0.27 g, 0.62 mmol) and palladium (II) acetate
(70
mg, 0.31 mmol) in anhydrous toluene (6 mL) was added N,N,N',N'-
tetramethylethylenediamine (0.29 mL, 2.48 mmol). The vial was sealed, heated
at
150 C for 22 h and then allowed to cool to 25 C. The reaction mixture was
diluted
with ethyl acetate, washed with water and brine, dried over Na2SO4, filtered
and
concentrated. The residue was purified on silica gel (gradient elution with 5%
ethyl
acetate/hexanes to 25% ethyl acetate/hexanes) to afford Cap-139, step a (669.7
mg,
59%) as a white solid. 1H NMR (CDC13, 500 MHz) 6 8.54 (d, J= 6.0 Hz, 1H), 8.22
(d, J= 9.0 Hz, 1H), 7.76 (d, J= 5.5 Hz, 1H), 7.41-7.39 (m, 1H), 7.13 (d, J=
2.0 Hz,
1H), 3.98 (s, 3H); Rt = 1.66 min (Cond.-D1); 90% homogenity index; LCMS: Anal.
Calc. for [M+H]+ C11H9N20: 185.07; found: 185.2.
Cap-139
Cap-139 was prepared from the basic hydrolysis of Cap-139, step a with 5N
NaOH according to the procedure described for Cap 138. 1H NMR (400 MHz,
DMSO-d6) 6 13.63 (v br s, 1H), 8.60 (d, J= 9.3 Hz, 1H), 8.45 (d, J= 5.6 Hz,
1H),
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7.95 (d, J= 5.9 Hz, 1H), 7.49 (d, J= 2.2 Hz, 1H), 7.44 (dd, J= 9.3, 2.5 Hz,
1H), 3.95
(s, 3H); Rt = 0.64 min (Cond.-D1); 90% homogenity index; LCMS: Anal. Calc. for
[M+H]+ C11H10NO3: 204.07; found: 204.05.
Cap-140
elN CI
CO2H
Cap-140, step a
-,0
40 '
N CI
ON
To a vigorously-stirred mixture of 1,3-dichloro-5-ethoxyisoquinoline (482
mg, 2.00 mmol; prepared according to the procedure in WO 2005/ 051410),
palladium (II) acetate (9 mg, 0.04 mmol), sodium carbonate (223 mg, 2.10 mmol)
and 1,5-bis(diphenylphosphino)pentane (35 mg, 0.08 mmol) in dry
dimethylacetamide (2 mL) at 25 C under nitrogen was added N,N,N',N'-
tetramethylethylenediamine (60 mL, 0.40 mmol). After 10 min, the mixture was
heated to 150 C, and then a stock solution of acetone cyanohydrin (prepared
from
457 [IL of acetone cyanohydrin in 4.34 mL DMA) was added in 1 mL portions over
18 h using a syringe pump. The mixture was then partitioned between ethyl
acetate
and water and the organic layer was separated, washed with brine, dried over
Na2SO4, filtered and concentrated. The residue was purified on silica gel
(gradient
elution with 10% ethyl acetate in hexanes to 40% ethyl acetate in hexanes) to
afford
Cap-140, step a (160 mg, 34%) as a yellow solid. Rt = 2.46 min (Cond.-MS-W2);
90% homogenity index; LCMS: Anal. Calc. for [M+H]+ C12H9C1N20: 233.05;
found: 233.08.
Cap-140
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Cap-140 was prepared by the acid hydrolysis of Cap-140, step a with 12N
HC1 as described in the procedure for the preparation of Cap 141, described
below.
Rt = 2.24 min (Cond.-MS-W2); 90% homogenity index; LCMS: Anal. Calc. for
[M+H]+ C12H11C1NO3: 252.04; found: 252.02.
Cap-141
elN F
CO2H
Cap-141, step a
40 '
N F
CN
Cap-141, step a was prepared from 1-bromo-3-fluoroisoquinoline (prepared
from 3-amino-1-bromoisoquinoline using the procedure outlined in J Med. Chem.
1970, 13, 613) as described in the procedure for the preparation of Cap-140,
step a
(vide supra). 1H NMR (500 MHz, CDC13) 6 8.35 (d, J= 8.5 Hz, 1H), 7.93 (d, J=
8.5 Hz, 1H), 7.83 (t, J= 7.63 Hz, 1H), 7.77-7.73 (m, 1H), 7.55 (s, 1H); Rt =
1.60 min
(Cond.-D1); 90% homogenity index; LCMS: Anal. Calc. for [M+H]+ C10H6FN2:
173.05; found: 172.99.
Cap-141
Cap-141, step a (83 mg, 0.48 mmol) was treated with 12N HC1 (3 mL) and
the resulting slurry was heated at 80 C for 16 h before it was cooled to room
temperature and diluted with water (3 mL). The mixture was stirred for 10 min
and
then filtered to afford Cap-141 (44.1 mg, 48%) as an off-white solid. The
filtrate was
diluted with dichloromethane and washed with brine, dried over Na2SO4, and
concentrated to afford additional Cap-141 (29.30 mg, 32%) which was
sufficiently
pure to be carried forward directly. 1H NMR (DMSO-d6, 500 MHz) 6 14.0 (br s,
1H), 8.59-8.57 (m, 1H), 8.10 (d, J= 8.5 Hz, 1H), 7.88-7.85 (m, 2H), 7.74-7.71
(m,
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1H); Rt= 1.33 min (Cond.-D1); 90% homogenity index; LCMS: Anal. Calc. for
[M+H]+ C10H7FN02: 192.05; found: 191.97.
Cap-142
o
c)
N
\
S 1\1
CO2H
Cap-142, step a
Br
lel A\I
ON
Cap-142, step a was prepared from 4-bromoisoquinoline N-oxide as
described in the two-step procedure for the preparation of Cap-138, steps b
and c. Rt
= 1.45 mm (Cond.-MS-W1); 90% homogenity index; LCMS: Anal. Calc. for
[M+H]+ C10H6BrN2: 232.97; found: 233.00.
Cap-142, step b
o
C)
N
S 1\1
ON
To an argon-degassed suspension of Cap-142, step a (116 mg, 0.50 mmol),
potassium phosphate tribasic (170 mg, 0.80 mmol), palladium (II) acetate (3.4
mg,
0.015 mmol) and 2-(dicyclohexylphosphino)biphenyl (11 mg, 0.03 mmol) in
anhydrous toluene (1 mL) was added morpholine (61 [IL, 0.70 mmol). The mixture
was heated at 100 C for 16 h, cooled to 25 C, filtered through diatomaceous
earth
(Celite ) and concentrated. Purification of the residue on silica gel
(gradient elution
with 10% to 70% ethyl acetate in hexanes) afforded Cap-142, step b (38 mg,
32%) as
a yellow solid which was carried forward directly. Rt = 1.26 min (Cond.-MS-
W1);
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90% homogenity index; LCMS: Anal. Calc. for [M+I-1]+ C14H14N30: 240.11; found:
240.13.
Cap-142
Cap-142 was prepared from Cap-142, step b with 5N sodium hydroxide as
described in the procedure for Cap 138. Rt = 0.72 min (Cond.-MS-W1); 90%
homogenity index; LCMS: Anal. Calc. for [M+I-1]+ C14H15N203: 259.11; found:
259.08.
Cap-143
r'o
NJ
co2H
Cap-143, step a
r'o
NJ
40 /N
Br
To a stirred solution of 3-amino-1-bromoisoquinoline (444 mg, 2.00 mmol) in
anhydrous dimethylformamide (10 mL) was added sodium hydride (60%, unwashed,
96 mg, 2.4 mmol) in one portion. The mixture was stirred at 25 C for 5 min
before
2-bromoethyl ether (90%, 250 uL, 2.00 mmol) was added. This mixture was
stirred
further at 25 C for 5 h and at 75 C for 72 h before it was cooled to 25 C,
quenched
Cap-143
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To a cold (-60 C) solution of Cap-143, step a (154 mg, 0.527 mmol) in
anhydrous tetrahydrofuran (5 mL) was added a solution of n-butyllithium in
hexanes
(2.5 M, 0.25 mL, 0.633 mmol). After 10 min, dry carbon dioxide was bubbled
into
the reaction mixture for 10 min before it was quenched with 1N HC1 and allowed
to
warm to 25 C. The mixture was then extracted with dichloromethane (3 x 30 mL)
and the combined organic extracts were concentrated in vacuo. Purification of
the
residue by reverse phase HPLC (Me0H/water/TFA) afforded Cap-143 (16 mg,
12%). Rt = 1.10 min (Cond.-MS-W1); 90% homogenity index; LCMS: Anal. Calc.
for [M+H]+ C14H15N203: 259.11; found: 259.08.
Cap-144
-.. ....-
N
CI
CO2H
Cap-144, step a
NO2
CI
40 '
N
a
1,3-Dichloroisoquinoline (2.75 g, 13.89 mmol) was added in small portions to
a cold (0 C) solution of fuming nitric acid (10 mL) and concentrated sulfuric
acid
(10 mL). The mixture was stirred at 0 C for 0.5 h before it was gradually
warmed to
C where it stirred for 16 h. The mixture was then poured into a beaker
containing
20 chopped ice and water and the resulting suspension was stirred for 1 h
at 0 C before
it was filtered to afford Cap-144, step a (2.73 g, 81%) as a yellow solid
which was
used directly. Rt = 2.01 min (Cond.-D1); 95% homogenity index; LCMS: Anal.
Calc. for [M+H]+ C9H5C12N202: 242.97; found: 242.92.
25 Cap-144, step b
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N/
SIN CI
CI
Cap-144, step a (0.30 g, 1.23 mmol) was taken up in methanol (60 mL) and
treated with platinum oxide (30 mg), and the suspension was subjected to Parr
hydrogenation at 7 psi H2 for 1.5 h before formalin (5 mL) and additional
platinum
oxide (30 mg) were added. The suspension was resubjected to Parr hydrogenation
at
45 psi H2 for 13 h before it was suction-filtered through diatomaceous earth
(Celite )
and concentrated down to 1/4 volume. Suction-filtration of the ensuing
precipitate
afforded the title compound as a yellow solid which was flash chromatographed
on
silica gel (gradient elution with 5% ethyl acetate in hexanes to 25% ethyl
acetate in
Cap-144, step c
',..N.,
00 '
N CI
CN
Cap-144, step c was prepared from Cap-144, step b according to the
procedure described for the preparation of Cap-139, step a. Rt= 2.19 min
(Cond.-
found: 232.03. HRMS: Anal. Calc. for [M+H]+ C12H11C1N3: 232.0642; found:
232.0631.
Cap-144
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Cap-144 was prepared according to the procedure described for Cap-141. Rt
= 2.36 min (Cond.-D1); 90%; LCMS: Anal. Calc. for [M+H]+ C12H12C1N202:
238.01; found: 238.09.
Caps-145 to -162
Caps-145 to 162 were prepared from the appropriate 1-chloroisoquinolines
according to the procedure described for the preparation of Cap-138 (Method A)
or
Cap-139 (Method B) unless noted otherwise as outlined below.
R( LC-
Cond.);
%
Cap # Cap Method Hydrolysis homogen
eity
index;
MS data
1.14 min
(Cond.-
MS-W1);
90%;
40 '
AI CI
LCMS:
Cap- CO2H Anal.
145 Prepared from B 12N HC1 Calc. for
[M+H]+
commercially available 1,3-
dichloroisoquinoline C10H7C1N
02:
208.02;
found:
208.00.
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1.40 min
(Cond.-
DO;
95%;
o
el " LCMS:
1\1
Cap- Anal.
co2H
146 A 5NNaOH Calc. for
Prepared from
+
commercially available 3-
[M+I-1]
hydroxyisoquinoline CiiHioN
03:
204.07;
found:
204.06.
0.87 min
(Cond.-
DO;
'a 95%;
el LCMS:
Cap- co2H Anal.
147 Prepared from B 5NNaOH Calc.
for
commercially available 1- [M+I-1]+
chloro-4- CiiHioN
hydroxyisoquinoline 03:
204.07;
found:
204.05.
o 40 0.70 min
Cap- co2H (Cond.-
148 Prepared from A 5NNaOH
DO;
commercially available 7- 95%;
hydroxyisoquinoline LCMS:
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Anal.
Calc. for
[M+H]+
CHHioN
03:
204.07;
found:
204.05.
0.70 min
(Cond.-
DO;
95%;
'o
C 40 LCMS:
Cap-
Anal.
co2H
149 A 5NNaOH Calc. for
Prepared from
[M+H]+
commercially available 5-
C1 1H10N
hydroxyisoquinoline
03:
204.07;
found:
204.05.
0.26 min
lel
. . = - - N TFA (Cond.-
DO;
o co2H
95%;
Cap- Prepared from 8-methoxy-
LCMS:
150 1-chloroisoquinoline, which A 12N HC1
Anal.
can be synthesized
Calc. for
following the procedure in [M+H]+
WO 2003/ 099274
CHHioN
03:
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204.07;
found:
204.04.
1.78 min
(Cond.-
DO;
'o
40 '
N CI 90%;
LCMS:
CO2H
Cap- Anal.
Prepared from 5-methoxy-
151 B 12N HC1 Calc. for
1,3-dichloroisoquinoline,
[M+H]+
which can be synthesized
CiiH9C1N
following the procedure in
03:
WO 2005 /051410.
238.03;
found:
238.09.
1.65 min
(Cond.-
DO;
o ei 95%;
WI ,N LCMS:
H
Cap- co2 Anal.
152 Prepared from B 12N HC1
Calc. for
commercially available 6- [M+H]+
methoxy-1,3- C11H9C1N
dichloroisoquinoline
03:
238.00;
found:
238.09.
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1.18 min
(Cond.-
Br MS-W1);
40 95%;
LCMS:
co2H
Cap- Anal.
Prepared from 4-
153 A 6N HC1 Calc. for
bromoisoquinoline, which
[M+H]+
can be synthesized
CioH7Br
following the procedure in
NO2:
WO 2003/ 062241
251.97;
found:
251.95.
0.28 min
(Cond.-
MS-W1);
40 90%;
F
co2H LCMS:
Cap- Prepared from 7-fluoro-1-
Anal.
154 chloroisoquinoline, which B 5N NaOH Calc. for
+
can be synthesized [M+H]
following the procedure in C1oH7FN
WO 2003/ 099274 02:
192.05;
found:
192.03.
0 0.59 min
ci
Cap- co2H (Cond.-
155 B 5NNaOH MS-W1);
Prepared from 1,7-
dichloroisoquinoline, which 90%;
LCMS:
can be synthesized
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following the procedure in Anal.
WO 2003/ 099274 Calc. for
[M+H]+
CioH2C1N
02:
208.02;
found:
208.00.
0.60 min
(Cond.-
MS-W1);
CI 090%;
1\1
co2H LCMS:
Cap- Prepared from 1,6- Anal.
156 dichloroisoquinoline, which B 5N NaOH
Calc. for
+
can be synthesized [M+H]
following the procedure in C10H2C1N
WO 2003/ 099274 02:
208.02;
found:
208.03.
1.49 min
ci (Cond.-
Di);
co2H 95%;
Cap-
Prepared from 1,4- LCMS:
157 B 12N HC1
dichloroisoquinoline, Anal.
which can be synthesized Calc. for
following the procedure in [M+H]+
WO 2003/ 062241 C10H12C1
NO:
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208.02;
found:
208.00.
0.69 min
(Cond.-
MS-W1);
ei
Si'l 90%;
LCMS:
co2H
Cap- Anal.
Prepared from 1,5-
/58 B 5N NaOH Calc. for
dichloroisoquinoline,
[M+I-1]+
which can be synthesized
CioH7C1N
following the procedure in
02:
WO 2003/ 099274
208.02;
found:
208.01.
0.41 min
(Cond.-
MS-W1);
F
0 i'l 90%;
LCMS:
co2H
Cap- Anal.
Prepared from 5-fluoro-1-
159 B 5N NaOH Calc. for
chloroisoquinoline,
[M+I-1]+
which can be synthesized
CioH7FN
following the procedure in
02:
WO 2003/ 099274
192.05;
found:
192.03.
Cap- F 0
1\1 B 5N NaOH 0.30 min
160 (Cond.-
co2H
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Prepared from 6-fluoro-1- MS-W1);
chloroisoquinoline, 90%;
which can be synthesized LCMS:
following the procedure in Anal.
WO 2003/ 099274 Calc. for
[M+I-1]+
CioH7FN
02:
192.05;
found:
192.03.
0.70 min
(Cond.
D1);
\N/
95%;
lel LCMS:
N CO2H
Cap- Anal.
Prepared from 4-
161Calc. for
bromoquinoline-2-
[M+I-1]+
carboxylic acid and
dimethylamine (DMSO, C12H13N2
100 C) 02:
217.10;
found:
217.06.
0.65 min
40 =
, N CO2H (Cond.-
Cap- Prepared from m-anisidine M3);
162 following the procedure 95%;
described in J. Hetero. LCMS:
Chem. 1993, 17 and Anal.
Heterocycles, 2003, 60, Calc. for
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953. [M+H]+
CHHioN
03:
204.07;
found:
203.94.
Cap-163
OH
1101 0OH
To a solution of 2-ketobutyric acid (1.0 g, 9.8 mmol) in diethylether (25 ml)
was added phenylmagnesium bromide (22 ml, 1M in THF) dropwise. The reaction
was stirred at ¨25 C under nitrogen for 17.5h. The reaction was acidified
with 1N
HC1 and the product was extracted with ethyl acetate (3 x 100 m1). The
combined
organic layer was washed with water followed by brine and dried over MgSO4.
After
Cap-164
SO
OH
---N
\
A mixture of 2-amino-2-phenylbutyric acid (1.5 g, 8.4 mmol), formaldehyde
(14 mL, 37% in water), 1N HC1 (10 mL) and 10% Pd/C (0.5 mg) in Me0H (40 mL)
was exposed to H2 at 50 psi in a Parr bottle for 42 h. The reaction was
filtered over
Celite and concentrated in vacuo, the residue was taken up in Me0H (36 mL) and
the
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product was purified with a reverse phase HPLC (Me0H/H20/TFA) to afford the
TFA salt of Cap-164 as a white solid (1.7 g). 1H NMR (DMSO-d6, 6 = 2.5 ppm,
500
MHz) 7.54-7.47 (m, 5H), 2.63 (m, 1H), 2.55 (s, 6H), 2.31 (m, 1H), 0.95 (app t,
J =
7.3 Hz, 3H).
Cap-165
0
400& OH
4111W
To a mixture of 2-amino-2-indanecarboxylic acid (258.6 mg, 1.46 mmol) and
formic acid (0.6 ml, 15.9 mmol) in 1,2-dichloroethane (7 ml) was added
formaldehyde (0.6 ml, 37% in water). The mixture was stirred at ¨25 C for 15
min
then heated at 70 C for 8h. The volatile component was removed in vacuo, and
the
residue was dissolved in DMF (14 mL) and purified by a reverse phase HPLC
(Me0H/H20/TFA) to afford the TFA salt of Cap-165 as a viscous oil (120.2 mg).
1H
NMR (DMSO-d6, 6 = 2.5 ppm, 500 MHz): 7.29-7.21 (m, 4 H), 3.61 (d, J = 17.4 Hz,
2H), 3.50 (d, J = 17.4 Hz, 2H), 2.75 (s, 6H). LC/MS: Anal. Calcd. for [M+H]+
C12H16NO2: 206.12; found: 206.07.
Cap-166a and -166b
is 0
OH Cap-166a: Diastereomer-1
Cap-166b: Diastereomer-2
Caps-166a and -166b were prepared from (1S, 45)-(+)-2-methy1-2,5-
diazabicyclo[2.2.1]heptane (2HBr) according to the method described for the
synthesis of Cap-7a and Cap-7b, with the exception that the benzyl ester
intermediate was separated using a semi-prep Chrialcel OJ column, 20 x 250 mm,
10
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p.m eluting with 85:15 heptane/ethanol mixture at 10 mL/min elution rate for
25 min.
Cap-166b: 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500 MHz): 7.45 (d, J = 7.3 Hz, 2H),
7.27-7.19 (m, 3H), 4.09 (s, 1H), 3.34 (app br s, 1H), 3.16 (app br s, 1H),
2.83 (d,
J=10.1 Hz, 1H), 2.71 (m, 2H), 2.46 (m, 1H), 2.27 (s, 3H), 1.77 (d, J = 9.8 Hz,
1H),
1.63 (d, J = 9.8 Hz, 1H). LC/MS: Anal. Calcd. for [M+H]+ C14H19N202: 247.14;
found: 247.11.
Cap-167
ON¨
OH
0
A solution of racemic Boc-1,3-dihydro-2H-isoindole carboxylic acid (1.0g,
3.8 mmol) in 20% TFA/CH2C12 was stirred at ¨25 C for 4h. All the volatile
component was removed in vacuo. A mixture of the resultant crude material,
formaldehyde (15 mL, 37% in water), 1N HC1 (10 mL) and 10% Pd/C (10 mg) in
Me0H was exposed to H2 (40 PSI) in a Parr bottle for 23 h. The reaction
mixture
was filtered over Celite and concentrated in vacuo to afford Cap-167 as a
yellow
foam (873.5 mg). 1H NMR (DMSO-d6, 6 = 2.5 ppm, 500 MHz) 7.59-7.38 (m, 4H),
5.59 (s, 1H), 4.84 (d, J = 14 Hz, 1H), 4.50 (d, J = 14.1 Hz, 1H), 3.07 (s,
3H). LC/MS:
Anal. Calcd. for [M+H]+ C10H12NO2: 178.09; found: 178.65.
Cap- 168
OH
0
NMe2
SO
Racemic Cap- 168 was prepared from racemic Boc-aminoindane-l-carboxylic
acid according to the procedure described for the preparation of Cap-167. The
crude
material was employed as such.
Cap- 169
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So
OH
N¨
I
A mixture of 2-amino-2-phenylpropanoic acid hydrochloride (5.0 g, 2.5
mmol), formaldehyde (15 ml, 37% in water), 1N HC1 (15 ml), and 10% Pd/C (1.32
g)
in Me0H (60 mL) was placed in a Parr bottle and shaken under hydrogen (55 PSI)
for 4 days. The reaction mixture was filtered through diatomaceous earth
(Celitec))
and concentrated in vacuo. The residue was taken up in Me0H and purified by
reverse phase prep-HPLC (Me0H/water/TFA) to afford the TFA salt of Cap-169 as
a
viscous semi-solid (2.1 g). 1H NMR (CDC13, 6 = 7.26 ppm, 500 MHz): 7.58-7.52
(m, 2 H), 7.39-7.33 (m, 3H), 2.86 (br s, 3H), 2.47 (br s, 3H), 1.93 (s, 3H).
LC/MS:
Anal. Calcd. for [M+H]+ C11H16NO2: 194.12; found: 194.12.
Cap-170
0 0
1?LOH
HN 0
r
0
To (S)-2-amino-2-(tetrahydro-2H-pyran-4-yl)acetic acid (505mg; 3.18mmol;
obtained from Astatech) in water (15m1) was added sodium carbonate (673mg;
6.35mmol), and the resultant mixture was cooled to 0 C and then methyl
chloroformate (0.26m1; 3.33mmol) was added dropwise over 5 minutes. The
reaction
was allowed to stir for 18 hours while allowing the bath to thaw to ambient
temperature. The reaction mixture was then partitioned between 1N HC1 and
ethyl
acetate. The organic layer was removed and the aqueous layer was further
extracted
with 2 additional portions of ethyl acetate. The combined organic layers were
washed with brine, dried over magnesium sulfate, filtered and concentrated in
vacuo
to afford Cap-170 a colorless residue. 1H NMR (500 MHz, DMSO-d6) 6 ppm 12.65
(1 H, br s), 7.44(1 H, d, J=8.24 Hz), 3.77 - 3.95 (3 H, m), 3.54(3 H, s), 3.11
- 3.26 (2
H, m), 1.82 - 1.95 (1 H, m), 1.41 - 1.55 (2 H, m), 1.21 - 1.39 (2 H, m);
LC/MS: Anal.
Calcd. for [M+H]+ C9H16N05: 218.1; found 218.1.
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Cap-171
Opy L
OLi
HN 0
f
0
A solution of methyl 2-(benzyloxycarbonylamino)-2-(oxetan-3-
ylidene)acetate (200 mg, 0.721 mmol; Il Farmaco (2001), 56, 609-613) in ethyl
acetate (7 ml) and CH2C12 (4.00 ml) was degassed by bubbling nitrogen for
10min.
Dimethyl dicarbonate (0.116 ml, 1.082 mmol) and Pd/C (20 mg, 0.019 mmol) were
then added, the reaction mixture was fitted with a hydrogen balloon and
allowed to
stir at ambient temperature overnight at which time TLC (95:5 CH2C12 / MeOH:
visulalized with stain made from lg Ce(NH4)2SO4, 6g ammonium molybdate, 6m1
sulfuric acid, and 100m1 water) indicated complete conversion. The reaction
was
filtered through celite and concentrated. The residue was purified via Biotage
(load
with dichloromethane on 25 samplet; elute on 25S column with dichloromethane
for
3CV then 0 to 5% Me0H / dichloromethane over 250m1 then hold at 5% Me0H /
dichloromethane for 250m1; 9m1 fractions). Collected fractions containing
desired
material and concentrated to 120mg (81%) of methyl 2-(methoxycarbonylamino)-2-
(oxetan-3-yl)acetate as a colorless oil. 1H NMR (500 MHz, CHLOROFORM-D) 6
ppm 3.29 - 3.40 (m, J=6.71 Hz, 1 H) 3.70 (s, 3 H) 3.74 (s, 3 H) 4.55 (t,
J=6.41 Hz, 1
H) 4.58 - 4.68 (m, 2 H) 4.67 -4.78 (m, 2 H) 5.31 (br s, 1 H). LC/MS: Anal.
Calcd.
for [M+H]+ C81-114N05: 204.2; found 204Ø
To methyl 2-(methoxycarbonylamino)-2-(oxetan-3-yl)acetate (50 mg, 0.246
mmol) in THF (2 mL) and water (0.5 mL) was added lithium hydroxide monohydrate
(10.33 mg, 0.246 mmol). The resultant solution was allowed to stir overnite at
ambient temperature. TLC (1:1 EA / Hex; Hanessian stain [1g Ce(NH4)2SO4, 6g
ammonium molybdate, 6m1 sulfuric acid, and 100m1 water]) indicated ¨10%
starting
material remaining. Added an additional 3mg LiOH and allowed to stir overnight
at
which time TLC showed no starting material remaining. Concentrated in vacuo
and
placed on high vac overnite providing 55mg lithium 2-(methoxycarbonylamino)-2-
(oxetan-3-yl)acetate as a colorless solid. 1H NMR (500 MHz, Me0D) 6 ppm 3.39 -
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3.47 (m, 1 H) 3.67 (s, 3 H) 4.28 (d, J=7.93 Hz, 1 H) 4.64 (t, J=6.26 Hz, 1 H)
4.68 (t,
J=7.02 Hz, 1 H) 4.73 (d, J=7.63 Hz, 2 H).
Cap-172
CI
OH
SI ____________________________________ /
N 0
Cap-1 72, step a
CI
o¨
z ____________________________________
sI / % 0
The following diazotization step was adapted from Barton, A.; Breukelman,
S. P.; Kaye, P. T.; Meakins, G. D.; Morgan, D. J. J. C. S. Perkin Trans I
1982,
159-164: A solution of NaNO2 (166 mg, 2.4 mmol) in water (0.6 mL) was added
slowly to a stirred, cold (0 C) solution of methyl 2-amino-5-ethy1-1,3-
thiazole-4-
carboxylate (186 mg, 1.0 mmol), CuSO4=5H20 (330 mg, 1.32 mmol), NaC1 (260 mg,
4.45 mmol) and H2SO4 (5.5 mL) in water (7.5 mL). The mixture was stirred at 0
C
for 45 min and allowed to warm up to room temperature where it stirred further
for 1
h before CuCl (118 mg) was added. This mixture was stirred further at room
temperature for 16 h before it was diluted with brine and extracted with ether
twice.
The organic layers were combined, dried over MgSO4 and concentrated to give
methyl 2-chloro-5-ethylthiazole-4-carboxylate (i.e. Cap-172, step a) (175 mg,
85%)
as an orange oil (80% pure) which was used directly in the next reaction. Rt =
1.99
min (Cond.-MD1); LC/MS: Anal. Calcd. for [M+H]+ C2H9C1NO2S: 206.01; found:
206.05.
Cap-172
To a solution of methyl 2-chloro-5-ethylthiazole-4-carboxylate (175 mg) in
THF/H20/Me0H (20 mL/ 3 mL/ 12 mL) was added LiOH (305 mg, 12.76 mmol).
The mixture was stirred at room temperature overnight before it was
concentrated
down and neutralized with 1N HC1 in ether (25 mL). The residue was extracted
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twice with ethyl acetate and the organic layers were combined, dried over
MgSO4
and evaporated to yield Cap-1 72 (60 mg, 74%) as a red solid which was used
without
further purification. 1H NMR (300 MHz, DMSO-d6) 6 ppm 13.03-13.42 (1 H, m),
3.16(2 H, q, J= 7.4 Hz), 1.23 (3 H, t, J= 7.5 Hz). Rt = 1.78 min (Cond.-MD1);
LC/MS: Anal. Calcd. for [M+H]+ C6H2C1NO2S: 191.99; found: 191.99.
Cap-1 73
SIO
N ____________________________________ µH0
Cap-1 73, step a
o¨
sI
N
The following diazotization step was adapted from Barton, A.; Breukelman,
S. P.; Kaye, P. T.; Meakins, G. D.; Morgan, D. J. J. C. S. Perkin Trans I
1982,
159-164: A solution of NaNO2 (150 mg, 2.17 mmol) in water (1.0 mL) was added
dropwise to a stirred, cold (0 C) solution of methyl 2-amino-5-ethy1-1,3-
thiazole-4-
carboxylate (186 mg, 1.0 mmol) in 50% H3P02 (3.2 mL). The mixture was stirred
at
0 C for 1 h and allowed to warm up to room temperature where it stirred
further for
2h. After recooling to 0 C, the mixture was treated slowly with a solution of
NaOH
(85 mg) in water (10 mL). The mixture was then diluted with saturated NaHCO3
solution and extracted twice with ether. The organic layers were combined,
dried
over MgSO4 and concentrated to give methyl 5-ethylthiazole-4-carboxylate (i.e.
Cap-1 73, step a) (134 mg, 78%) as an orange oil (85% pure) which was used
directly
in the next reaction. Rt = 1.58 min (Cond.-MD1); LC/MS: Anal. Calcd. for
[M+H]+
C21-110NO2S: 172.05; found: 172.05.
Cap-1 73
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To a solution of methyl 5-ethylthiazole-4-carboxylate (134 mg) in
THF/H20/Me0H (18 mL/ 2.7 mL/ 11 mL) was added LiOH (281 mg, 11.74 mmol).
The mixture was stirred at room temperature overnight before it was
concentrated
down and neutralized with 1N HC1 in ether (25 mL). The residue was extracted
twice with ethyl acetate and the organic layers were combined, dried over
MgSO4
and evaporated to yield Cap-173 (90 mg, 73%) as an orange solid which was used
without further purification. 1H NMR (300 MHz, DMSO-d6) 6 ppm 12.74-13.04 (1
H, m), 3.20 (2 H, q, J= 7.3 Hz), 1.25 (3 H, t, J= 7.5 Hz). Rt = 1.27 min
(Cond.-
MD1); LC/MS: Anal. Calcd. for [M+H]+ C6H8NO2S: 158.03; found: 158.04.
Cap-174
1
I N..roEi
o
Cap-174, step a
......õ.......,so3cF3
I
Nr
o
Triflic anhydride (5.0 g, 18.0 mmol) was added dropwise to a cold (0 C)
solution of methyl 3-hydroxypicolinate (2.5 g, 16.3 mmol) and TEA (2.5 mL,
18.0
mmol) in CH2C12 (80 mL). The mixture was stirred at 0 C for lh before it was
allowed to warm up to room temperature where it stirred for an additional 1 h.
The
mixture was then quenched with saturated NaHCO3 solution (40 mL) and the
organic
layer was separated, washed with brine, dried over MgSO4 and concentrated to
give
methyl 3-(trifluoromethylsulfonyloxy)picolinate (i.e. Cap-174, step a) (3.38
g, 73%)
as a dark brown oil (>95% pure) which was used directly without further
purification. 1H NMR (300 MHz, CDC13) 6 ppm 8.72-8.79(1 H, m), 7.71(1 H, d, J
= 1.5 Hz), 7.58-7.65 (1 H, m), 4.04 (3 H, s). Rt = 1.93 min (Cond.-MD1);
LC/MS:
Anal. Calcd. for [M+H]+ C8H2F3NO5S: 286.00; found: 286.08.
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Cap-174
To a solution of methyl 3-(trifluoromethylsulfonyloxy)picolinate (570 mg, 2.0
mmol) in DMF (20 mL) was added LiC1 (254 mg, 6.0 mmol),
tributyl(vinyl)stannane
(761 mg, 2.4 mmol) and bis(triphenylphosphine)palladium dichloride (42 mg,
0.06
mmol). The mixture was heated at 100 C overnight before a saturated solution
of
KF (20 mL) was added to the reaction mixture at room temperature. This mixture
was stirred for 4 h before it was filtered through diatomaceous earth
(Celitec)) and the
pad was washed with ethyl acetate. The aqueous phase of the filtrate was then
separated and concentrated down in vacuo. The residue was treated with 4N HC1
in
dioxanes (5 mL) and the resulting mixture was extracted with methanol,
filtered and
evaporated to afford Cap-1 74 (260 mg) as a green solid which was slightly
contaminated with inorganic salts but was used without further purification.
1H
NMR (300 MHz, DMSO-d6) 6 ppm 8.21 (1 H, d, J= 3.7 Hz), 7.81-7.90 (1 H, m),
7.09 (1 H, dd, J= 7.7, 4.8 Hz), 6.98 (1 H, dd, J= 17.9, 11.3 Hz), 5.74 (1 H,
dd, J=
17.9, 1.5 Hz), 5.20(1 H, d, J= 11.0 Hz). Rt = 0.39 min (Cond.-MD1); LC/MS:
Anal.
Calcd. for [M+H]+ C8H8NO2: 150.06; found: 150.07.
Cap-175
.r0H
N
0
Cap-1 75, step a
1
I
0
N
0
To a solution of methyl 3-(trifluoromethylsulfonyloxy)picolinate (i.e. Cap
173, step a) (570 mg, 2.0 mmol), an intermediate in the preparation of Cap-1
74, in
DMF (20 mL) was added LiC1 (254 mg, 6.0 mmol), tributyl(vinyl)stannane (761
mg,
2.4 mmol) and bis(triphenylphosphine)palladium dichloride (42 mg, 0.06 mmol).
The mixture was heated at 100 C for 4 h before the solvent was removed in
vacuo.
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The residue was taken up in acetonitrile (50 mL) and hexanes (50 mL) and the
resulting mixture was washed twice with hexanes. The acetonitrile layer was
then
separated, filtered through Celite, and evaporated. Purification of the
residue by
flash chromatography on a Horizon instrument (gradient elution with 25% ethyl
acetate in hexanes to 65% ethyl acetate in hexanes) afforded methyl 3-
vinylpicolinate
(i.e. Cap-1 75, step a) (130 mg, 40%) as a yellow oil. 1H NMR (300 MHz, CDC13)
6
ppm 8.60 (1 H, dd, J= 4.6, 1.7 Hz), 7.94 (1 H, d, J= 7.7 Hz), 7.33-7.51 (2 H,
m),
5.72 (1 H, d, J= 17.2 Hz), 5.47(1 H, d, J= 11.0 Hz), 3.99(3 H, s). Rt = 1.29
min
(Cond.-MD1); LC/MS: Anal. Calcd. for [M+H]+ C9H10NO2: 164.07; found: 164.06.
Cap-1 75, step b
0
N \
o
Palladium on carbon (10%, 25 mg) was added to a solution of methyl 3-
vinylpicolinate (120 mg, 0.74 mmol) in ethanol (10 mL). The suspension was
stirred
at room temperature under an atmosphere of hydrogen for 1 h before it was
filtered
through Celite and the pad of diatomaceous earth (Celite ) was washed with
methanol. The filtrate was concentrated down to dryness to yield methyl 3-
ethylpicolinate (i.e. Cap-1 75, step b) which was taken directly into the next
reaction.
Rt = 1.15 min (Cond.-MD1); LC/MS: Anal. Calcd. for [M+H]+ C9H12NO2: 166.09;
found: 166.09.
Cap-1 75
To a solution of methyl 3-ethylpicolinate in THF/H20/Me0H (5 mL/ 0.75
mL/ 3 mL) was added LiOH (35 mg, 1.47 mmol). The mixture was stirred at room
temperature for 2 d before additional LiOH (80 mg) was added. After an
additional
24 h at room temperature, the mixture was filtered and the solvent was removed
in
vacuo. The residue was then treated with 4N HC1 in dioxanes (5 mL) and the
resulting suspension was concentrated down to dryness to yield Cap-1 75 as a
yellow
1
solid which was used without further purification. H NMR (300 MHz, DMSO-d6) 6
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ppm 8.47 (1 H, dd, J= 4.8, 1.5 Hz), 7.82-7.89 (1 H, m), 7.53 (1 H, dd, J= 7.7,
4.8
Hz), 2.82 (2 H, q, J= 7.3 Hz), 1.17 (3 H, t, J= 7.5 Hz). Rt = 0.36 min (Cond.-
MD1);
LC/MS: Anal. Calcd. for [M+H]+ C8H10NO2: 152.07; found: 152.10.
Cap-176
HO 00
FXN 0
H
F
Cap-176, step a
0 0
....6X 0 40
NO
0 H
---0
A solution of 1,4-dioxaspiro[4.5]decan-8-one (15 g, 96 mmol) in Et0Ac (150
mL) was added to a solution of methyl 2-(benzyloxycarbonylamino)-2-
(dimethoxyphosphoryl)acetate (21.21 g, 64.0 mmol) in 1,1,3,3-
tetramethylguanidine
(10.45 mL, 83 mmol) and Et0Ac (150 mL). The resulting solution was the stirred
at
ambient temperature for 72 h and then it was diluted with Et0Ac (25 mL). The
organic layer was washed with 1N HC1 (75 mL), H20 (100 mL) and brine (100 mL),
dried (MgSO4), filtered and concentrated. The residue was purified via Biotage
(5%
to 25 % Et0Ac/Hexanes; 300g column). The combined fractions containing the
product were then concentrated under vacuum and the residue was re-
crystallized
from hexanes/Et0Ac to give white crystals that corresponded to methyl 2-
(benzyloxycarbonylamino)-2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate (6.2 g)
1H
NMR (400 MHz, CDC13-d) 6 ppm 7.30 - 7.44 (5 H, m), 6.02 (1 H, br. s.), 5.15 (2
H,
s), 3.97 (4 H, s), 3.76 (3 H, br. s.), 2.84 - 2.92 (2 H, m), 2.47 (2 H, t,
J=6.40 Hz),
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1.74 - 1.83 (4 H, m). LC (Cond. OL1): Rt = 2.89 min. LC/MS: Anal. Calcd. For
[M+Na]+ C19H23NNa06: 745.21; found: 745.47.
Cap 176, step b
0 0 0
N .LO lei
0,0XH
_.--C)
Ester Cap 176, step b was prepared from alkene Cap 176, step a according to
the method of Burk, M. J.; Gross, M. F. and Martinez J. P. (J. Am. Chem. Soc.,
1995, 117, 9375-9376 and references therein): A 500 mL high-pressure bottle
was
charged with alkene Cap 176, step a (3.5 g, 9.68 mmol) in degassed Me0H (200
mL)
under a blanket of N2. The solution was then charged with (-)-1,2-Bis((2S,5S)-
2,5-
dimethylphospholano)ethane(cyclooctadiene)rhodium (I) tetrafluoroborate (0.108
g,
0.194 mmol) and the resulting mixture was flushed with N2 (3x) and charged
with H2
(3x). The solution was shaken vigorously under 70 psi of H2 at ambient
temperature
for 72 h. The solvent was removed under reduced pressure and the remaining
residue
was taken up in Et0Ac. The brownish solution was then filtered through a plug
of
Silica Gel and eluted with Et0Ac. The solvent was concentrated under vacuum to
afford a clear oil corresponding to ester Cap 176, step b (3.4 g) .1H NMR (500
MHz,
CDC13-d) 6 ppm 7.28 - 7.43 (5 H, m), 5.32 (1 H, d, J=9.16 Hz), 5.06 - 5.16 (2
H, m),
4.37 (1 H, dd, J=9.00, 5.04 Hz), 3.92 (4 H, t, J=3.05 Hz), 3.75 (3 H, s), 1.64
- 1.92 (4
H, m), 1.37 - 1.60 (5 H, m). LC (Cond. OL1): Rt = 1.95 mm. LC/MS: Anal. Calcd.
For [M+H]+ Ci9H26N06: 364.18; found: 364.27.
Cap 176, step c
0 0 0
JOXN 0 =
H
0
Ester Cap 176, step b (4.78 g, 13.15 mmol) was dissolved in THF (15 mL)
followed by sequential addition of water (10 mL), glacial acetic acid (26.4
mL, 460
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mmol) and dichloroacetic acid (5.44 mL, 65.8 mmol). The resulting mixture was
stirred for 72 h at ambient temperature, and the reaction was quenched by slow
addition of solid Na2CO3with vigorous stirring until the release of gas was no
longer
visible. Crude product was extracted into 10% ethyl acetate-dichloromethane
and the
organic layers were combined, dried (MgSO4) filtered and concentrated. The
resulting residue was purified via Biotage (0 to 30% Et0Ac/Hex; 25 g column)
to
afford ketone Cap 176, step c (3.86g) as a clear oil. 1H NMR (400 MHz, CDC13-
d) 6
ppm 7.28 - 7.41 (5 H, m), 5.55 (1 H, d, J=8.28 Hz), 5.09 (2 H, s), 4.46 (1 H,
dd,
J=8.16, 5.14 Hz), 3.74 (3 H, s), 2.18 -2.46 (5 H, m), 1.96 - 2.06 (1 H, m),
1.90 (1 H,
ddd, J=12.99, 5.96, 2.89 Hz), 1.44 - 1.68 (2 H, m, J=12.36, 12.36, 12.36,
12.36, 4.77
Hz). LC (Cond. OL1): Rt = 1.66 min. LC/MS: Anal. Calcd. For [M+Na]+
C12H21NNa05: 342.13; found: 342.10.
Cap 176, step d
Fd
:10 0
N 0
H
F
Deoxo-Fluor (3.13 mL, 16.97 mmol) was added to a solution of ketone Cap
176, step c (2.71 g, 8.49 mmol) in CH2C12 (50 mL) followed by addition of a
catalytic ammount of Et0H (0.149 mL, 2.55 mmol). The resulting yellowish
solution was stirred at rt overnight. The reaction was quenched by addition of
sat.
aq. NaHCO3 (25 mL) and the mixture was extracted with Et0Ac (3X75 mL)). The
combined organic layers were dried (MgSO4), filtered and dried to give a
yellowish
oil. The residue was purified via Biotage chromatography (2% to 15% Et0Ac/Hex;
90g column) and a white solid corresponding to the difluoro amino acid
dilforide
Cap 176, step d (1.5 g) was recovered. 1H NMR (400 MHzõ CDC13-d) 6 ppm 7.29 -
7.46 (5 H, m), 5.34 (1 H, d, J=8.28 Hz), 5.12 (2 H, s), 4.41 (1 H, dd, J=8.66,
4.89
Hz), 3.77 (3 H, s), 2.06 -2.20 (2 H, m), 1.83 - 1.98 (1 H, m), 1.60 - 1.81 (4
H, m),
1.38 - 1.55 (2 H, m). 19F NMR (376 MHz, CDC13-d) 6 ppm -92.15 (1 F, d,
J=237.55
Hz), -102.44 (1 F, d, J=235.82 Hz). LC (Cond. OL1): Rt = 1.66 min. LC/MS:
Anal. Calcd. For [2M+Na]+ C34H42F4N2Na08: 705.28; found: 705.18.
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Cap 176, step e
0 0
NH
F----i&C2
F
Difluoride Cap 176, step d (4 g, 11.72 mmol) was dissolved in Me0H (120
mL) and charged with Pd/C (1.247 g, 1.172 mmol). The suspension was flushed
with
N2 (3x) and the reaction mixture was placed under 1 atm of H2 (balloon). The
mixture was stirred at ambient temperature for 48 h. The suspension was then
filtered though a plug of Celite and concentrated under vacuum to give an oil
that
corresponded to amino acid Cap 176, step e (2.04 g) and that was used without
further purification. 1H NMR (400 MHz, DMSO-d6) 6 ppm 3.62 (3 H, s), 3.20 (1
H,
d, J=5.77 Hz), 1.91 - 2.09 (2 H, m), 1.50 - 1.88 (7 H, m), 1.20 - 1.45 (2 H,
m). 19F
NMR (376 MHz, DMSO-d6) 6 ppm -89.39 (1 F, d, J=232.35 Hz), -100.07 (1 F, d,
J=232.35 Hz). 13C NMR (101 MHz, DMSO-d6) 6 ppm 175.51(1 C, s), 124.10 (1 C,
t, J=241.21, 238.90 Hz), 57.74 (1 C, s), 51.39 (1 C, s), 39.23 (1 C, br. s.),
32.02 -
33.83 (2 C, m), 25.36 (1 C, d, J=10.02 Hz), 23.74 (1 C, d, J=9.25 Hz). LC
(Cond.
0L2): Rt = 0.95 min. LC/MS: Anal. Calcd. For [2M+H]+ C18I-131F4N202: 415.22;
found: 415.40.
Cap 176, step f
0 00____CH
N)L0
F
F
Methyl chloroformate (1.495 mL, 19.30 mmol) was added to a solution of
amino acid Cap 176, step e (2 g, 9.65 mmol) and DIEA (6.74 mL, 38.6 mmol) in
CH2C12(100 mL). The resulting solution was stirred at rt for 3 h and volatiles
were
removed under reduced pressure. The residue was purified via Biotage (0% to
20%
Et0Ac/Hex; 90g column). A clear oil that solidified upon standing under vacuum
and corresponding to carbamate Cap-176, step f(2.22 g) was recovered. 1H NMR
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(500 MHz, CDC13-d) 6 ppm 5.27 (1 H, d, J=8.55 Hz), 4.39 (1 H, dd, J=8.85, 4.88
Hz), 3.77 (3 H, s), 3.70 (3 H, s), 2.07 - 2.20 (2 H, m), 1.84 - 1.96 (1 H, m),
1.64 -
1.82 (4 H, m), 1.39 - 1.51 (2 H, m). 19F NMR (471 MHz, CDC13-d) 6 ppm -92.55
(1
F, d, J=237.13 Hz), -102.93 (1 F, d, J=237.12 Hz). 13C NMR (126 MHz, CDC13-d)
6
ppm 171.97 (1 C, s), 156.69(1 C, s), 119.77- 125.59(1 C, m), 57.24(1 C, br.
s.),
52.48 (1 C, br. s.), 52.43 (1 C, s), 39.15 (1 C, s), 32.50 - 33.48 (2 C, m),
25.30 (1 C,
d, J=9.60 Hz), 24.03 (1 C, d, J=9.60 Hz). LC (Cond. OL1): Rt = 1.49 min.
LC/MS:
Anal. Calcd. For [M+Na]+ CiiHi7F2NNa04: 288.10; found: 288.03.
Cap-176
A solution of LiOH (0.379 g, 15.83 mmol) in Water (25 mL) was added to a
solution of carbamate Cap-176, step f (2.1 g, 7.92 mmol) in THF (75 mL) and
the
resulting mixture was stirred at ambient temperature for 4 h. THF was removed
under
vacuum and the remaining aqueous phase was acidified with 1N HC1 solution (2
mL)
and then extracted with Et0Ac (2 X 50 mL). The combined organic layers were
dried
(MgSO4), filtered and concentrated to give a white foam corresponding to Cap-
176
(1.92 g). 1H NMR (400 MHz, DMSO-d6) 6 ppm 12.73 (1 H, s), 7.50(1 H, d, J=8.78
Hz), 3.97 (1 H, dd, J=8.53, 6.02 Hz), 3.54 (3 H, s), 1.92 - 2.08 (2 H, m),
1.57 - 1.90 (5
H, m), 1.34 - 1.48 (1 H, m), 1.27 (1 H, qd, J=12.72, 3.26 Hz). 19F NMR (376
MHz,
DMSO-d6) 6 ppm -89.62 (1 F, d, J=232.35 Hz), -99.93 (1 F, d, J=232.35 Hz). LC
(Cond. 0L2): Rt = 0.76 min. LC/MS: Anal. Calcd. For [M-H]+ C10H14F2N04:
250.09; found: 250.10.
BIOLOGICAL ACTIVITY
An HCV Replicon assay was utilized in the present disclosure, and was
prepared, conducted and validated as described in commonly owned
PCT/US2006/022197 and in O'Boyle et. al. Antimicrob Agents Chemother. 2005
Apr;49(4):1346-53. Assay methods incorporating luciferase reporters have also
been
used as described (Apath.com).
HCV-neo replicon cells and replicon cells containing resistance substitutions
in the NS5A region were used to test the currently described family of
compounds.
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The compounds were determined to have differing degrees of reduced inhibitory
activity on cells containing mutations vs. the corresponding inhibitory
potency
against wild-type cells. Thus, the compounds of the present disclosure can be
effective in inhibiting the function of the HCV NS5A protein and are
understood to
be as effective in combinations as previously described in application
PCT/US2006/022197 and commonly owned WO/04014852. It should be understood
that the compounds of the present disclosure can inhibit multiple genotypes of
HCV.
Table 2 shows the EC50 (Effective 50% inhibitory concentration) values of
representative compounds of the present disclosure against the HCV lb
genotype. In
one embodiment, compounds of the present disclosure are inhibitory versus la,
lb,
2a, 2b, 3a, 4a, and 5a genotypes. EC50 values against HCV lb are as follows A
(10-
350 nM); B (1-9.9 nM); C (0.1-0.99 nM); D (0.0006-0.099 nM).
Compound lb EC50 Range Name
No. (nM)
OL-1 B (1R)-2-((2S)-2-(4-(4-(4-(2-((2S)-142R)-2-
(dimethylamino)-2-phenylacety1)-2-
pyrrolidiny1)-1H-imidazol-5-y1)phenoxy)
pheny1)-1H-imidazol-2-y1)-1-pyrrolidiny1)-
N,N-dimethyl-2-oxo-1-phenylethanamine
OL-2 9.1 B (1R)-2-((2S)-2-(4-(4-(4-(2-((2S)-142R)-2-
hydroxy-2-phenylacety1)-2-pyrrolidiny1)-1H-
imidazol-5-y1)phenoxy)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethanol
OL-3 B dimethyl (oxybis(4,1-phenylene-1H-
imidazole-4,2-diy1(2S)-2,1-
pyrrolidinediy1((1R)-2-oxo-1-phenyl-2,1-
ethanediy1)))biscarbamate
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Compound lb EC50 Range Name
No. (nM)
OL-4 0.07 D (1R)-2-((2S)-2-(4-(3-(4-(2-((2S)-142R)-2-
(dimethylamino)-2-phenylacety1)-2-
pyrrolidiny1)-1H-imidazol-4-y1)phenoxy)
pheny1)-1H-imidazol-2-y1)-1-pyrrolidiny1)-
N,N-dimethyl-2-oxo-1-phenylethanamine
OL-5 80 A (1R)-2-((2S)-2-(4-(3-(4-(2-((2S)-142R)-2-
hydroxy-2-phenylacety1)-2-pyrrolidiny1)-1H-
imidazol-4-y1)phenoxy)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethanol
OL-6 D methyl ((lR)-242S)-2-(4-(3-(4-(242S)-1-
((2R)-2-((methoxycarbonyl)amino)-2-
phenylacety1)-2-pyrrolidinyl)-1H-imidazol-
4-y1)phenoxy)phenyl)-1H-imidazol-2-y1)-1-
pyrrolidinyl)-2-oxo-1-phenylethyl)carbamate
OL-7 D (1R)-2-((2S)-2-(4-(4-((4-(2-((2S)-1-((2R)-2-
(dimethylamino)-2-phenylacety1)-2-
pyrrolidiny1)-1H-imidazol-4-y1)benzyl)oxy)
pheny1)-1H-imidazol-2-y1)-1-pyrrolidiny1)-
N,N-dimethyl-2-oxo-1-phenylethanamine
OL-8 D methyl ((lR)-2-((2S)-2-(4-(4-((4-(2-((2S)-1-
((2R)-2-((methoxycarbonyl)amino)-2-
phenylacety1)-2-pyrrolidinyl)-1H-imidazol-
4-y1)benzyl)oxy)phenyl)-1H-imidazol-2-y1)-
1-pyrrolidinyl)-2-oxo-1-
phenylethyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
OL-9 D (1R)-2-((2 S)-2-(4-(4-(2-(4-(2-((2 S)-1 -((2R)-
2-(dimethylamino)-2-phenylac ety1)-2-
pyrrolidiny1)-1H-imidazol-5-y1)phenyl)ethyl)
pheny1)-1H-imidazol-2-y1)-1-pyrrolidiny1)-
N,N-dimethyl-2-oxo-l-phenylethanamine
OL-10 D (1R,1'R)-2,2'-(1,2-ethanediylbis(4,1-
phenylene-1H-imidazole-4,2-diy1(2S)-2,1-
pyrrolidinediy1))bis(2-oxo-1-phenylethanol)
OL-11 D dimethyl (1,2-ethanediylbis(4,1-phenylene-
1H-imidazole-4,2-diy1(2S)-2,1-
pyrrolidinediy1 ((1R)-2-oxo-1-phenyl-2,1-
ethanediy1))) biscarbamate
OL-12 D N',N'"-(1,2-ethanediylbis(4,1-phenylene-1H-
imidazole-4,2-diy1(2S)-2,1-pyrrolidinediy1
((1R)-2-oxo-1-phenyl-2,1-ethanediy1)))bis(1-
ethylurea)
OL-13 D 1-cyc lopenty1-3 -((1R)-2-((2 S)-2-(4-(4-(2-(4-
(2-((2 S)-1 -((2R)-2-((cyclop entylc arb amoyl)
amino)-2-phenylacety1)-2-pyrrolidiny1)-1H-
imidazol-5-y1)phenyl)ethyl)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethyl)urea
OL-14 C (1R)-242S)-2-(4-(44(4-(242S)-142R)-2-
(dimethylamino)-2-phenylacety1)-2-
pyrrolidiny1)-1H-imidazol-5-y1)benzyl)oxy)
methyl)pheny1)-1H-imidazol-2-y1)-1-
pyrrolidiny1)-N,N-dimethyl-2-oxo-l-
phenylethanamine
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Compound lb EC50 Range Name
No. (nM)
OL-15 1.1 B (1R)-242S)-2-(4-(44(4-(242S)-142R)-2-
hydroxy-2-phenylacety1)-2-pyrrolidiny1)-1H-
imidazol-5-y1)benzyl)oxy)methyl)pheny1)-
1H-imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethanol
OL-16 D dimethyl (oxybis (methylene-4,1-phenylene-
1H-imidazole-4,2-diy1(2 S)-2,1-
pyrrolidinediyl ((1R)-2-oxo-1-phenyl-2,1-
ethanediy1))) biscarbamate
OL-17 D 1-methy1-3-((1R)-2-((2S)-2-(4-(4-(((4-(2-
((2S)-142R)-2-((methylcarbamoyl)amino)-
2-phenylacety1)-2-pyrrolidinyl)-1H-
imidazol-5-y1)benzyl)oxy)methyl)phenyl)-
1H-imidazol-2-y1)-1-pyrrolidinyl)-2-oxo-1-
phenylethyl)urea
OL-18 D 1-ethy1-3-((1R)-2-((2S)-2-(4-(4-(((4-(2-
((2S)-142R)-2-((ethylcarbamoyl)amino)-2-
phenylac ety1)-2-pyrrolidiny1)-1H-imidazol-
-yl)benzyl)oxy)methyl)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethyl)urea
OL-19 B 1-cyclopenty1-3-((lR)-2-((2S)-2-(4-(4-(((4-
(2-((2S)-1-((2R)-2-((cyclopentylcarbamoyl)
amino)-2-phenylacety1)-2-pyrrolidiny1)-1H-
imidazol-5-y1)benzyl)oxy)methyl)pheny1)-
1H-imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethyl)urea
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Compound lb EC50 Range Name
No. (nM)
OL-20 C (1R)-2-((2S)-2-(4-(34(4-(242S)-142R)-2-
(dimethylamino)-2-phenylacety1)-2-
pyrrolidiny1)-1H-imidazol-4-y1)benzyl)oxy)
methyl)pheny1)-1H-imidazol-2-y1)-1-
pyrrolidiny1)-N,N-dimethyl-2-oxo-1-
phenylethanamine
OL-21 C (1R)-242S)-2-(4-(34(4-(242S)-142R)-2-
hydroxy-2-phenylacety1)-2-pyrrolidiny1)-1H-
imidazol-4-y1)benzyl)oxy)methyl)pheny1)-
1H-imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethanol
OL-22 D (methyl ((1R)-242S)-2-(4-(34(4-(2-((2S)-
142R)-2-((methoxycarbonyl)amino)-2-
phenylacety1)-2-pyrrolidinyl)-1H-imidazol-
4-y1)benzyl)oxy)methyl)phenyl)-1H-
imidazol-2-y1)-1-pyrrolidinyl)-2-oxo-1-
phenylethyl)carbamate
OL-23 C 1-methy1-3-((1R)-2-((2S)-2-(4-(3-(((4-(2-
((2S)-1-((2R)-2-((methylcarbamoyl)amino)-
2-phenylacety1)-2-pyrrolidiny1)-1H-
imidazol-4-y1)benzyl)oxy)methyl)pheny1)-
1H-imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethyl)urea
OL-24 D 1-ethy1-341R)-2-((2S)-2-(4-(34(4-(2-
((2S)-142R)-2-((ethylcarbamoyl)amino)-2-
phenylacety1)-2-pyrrolidiny1)-1H-imidazol-
4-y1)benzyl)oxy)methyl)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethyl)urea
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Compound lb EC50 Range Name
No. (nM)
OL-25 D 1-cyclopenty1-3-((lR)-2-((2S)-2-(4-(3-(((4-
(2-((2S)-1-((2R)-2-((cyclopentylcarbamoyl)
amino)-2-phenylacety1)-2-pyrrolidiny1)-1H-
imidazol-4-y1)benzyl)oxy)methyl)pheny1)-
1H-imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethyl)urea
D-1 D dimethyl (1,1':4',1"-terpheny1-4,4"-
diylbis(1H-imidazole-4,2-diy1(2S)-2,1-
pyrrolidinediyl ((1R)-2-oxo-1-phenyl-2,1-
ethanediy1))) biscarbamate
D-2 C (1R)-242S)-2-(4-(4"-(242S)-142R)-2-
(dimethylamino)-2-phenylacety1)-2-
pyrrolidiny1)-1H-imidazol-5-y1)-1,1':4',1"-
terphenyl-4-y1)-1H-imidazol-2-y1)-1-
pyrrolidiny1)-N,N-dimethyl-2-oxo-1-
phenylethanamine
0.006 D methyl ((1S)-1-(((2S)-2-(4-(4-((2-((2S)-1-
((2S)-2-((methoxycarbonyl)amino)-3-
methylbutanoy1)-2-pyrrolidiny1)-4,5-dihydro-
D-3 1H-naphtho[1,2-d]imidazol-7-
yl)ethynyl)pheny1)-1H-imidazol-2-y1)-1-
pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
methyl ((lR)-242S)-2-(4-(44242S)-1-
((2R)-2-((methoxycarbonyl)amino)-2-
phenylacety1)-2-pyrrolidiny1)-4,5-dihydro-
D-4
1H-naphtho[1,2-d]imidazol-7-
yl)ethynyl)pheny1)-1H-imidazol-2-y1)-1-
pyrrolidiny1)-2-oxo-1-phenylethyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
D methyl ((1 R)-242S)-2-(744-(242S)-1-
((2R)-2-((methoxycarbonyl)amino)-2-
phenylacety1)-2-pyrrolidiny1)-1H-imidazol-
D-5
4-yl)phenyl)ethyny1)-1H-naphtho[1,2-
d]imidazol-2-y1)-1-pyrrolidiny1)-2-oxo-1-
phenylethyl)carbamate
D methyl ((1S)-1-(((2S)-2-(4-(4-((2-((2S)-1-
((2S)-2-((methoxycarbonyl)amino)-3-
methylbutanoy1)-2-pyrrolidiny1)-1H-
D-6 naphtho[1,2-d]imidazol-7-
yl)ethynyl)pheny1)-1H-imidazol-2-y1)-1-
pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
D methyl ((1S)-1-(((1R,3S,5R)-3-(4-(442-
((1R,3S,5R)-242S)-2-
((methoxycarbonyl)amino)-3-
D methylbutanoy1)-2-azabicyclo[3.1.0]hex-3-
-7
y1)-1H-naphtho[1,2-d]imidazol-7-
y1)ethynyl)phenyl)-1H-imidazol-2-y1)-2-
azabicyclo[3.1.0]hex-2-y1)carbonyl)-2-
methylpropyl)carbamate
D
methyl ((1R)-2-((lR,3S,5R)-3-(7-((4-(2-
((1R,3S,5R)-242R)-2-
((methoxycarbonyl)amino)-2-phenylacety1)-
D-8 0.005 2-azabicyclo[3.1.0]hex-3-y1)-1H-imidazol-4-
yl)phenyl)ethyny1)-1H-naphtho[1,2-
d]imidazol-2-y1)-2-azabicyclo[3.1.0]hex-2-
y1)-2-oxo-1-phenylethyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
D methyl ((1S)-1-(((1R,3S,5R)-3-(7-((2-
((1R,3S,5R)-242S)-2-
((methoxycarbonyl)amino)-3-
D methylbutanoy1)-2-azabicyclo[3.1.0]hex-3-
-9
y1)-4,5-dihydro-1H-naphtho[1,2-d]imidazol-
7-yl)ethyny1)-4,5-dihydro-1H-naphtho[1,2-
d]imidazol-2-y1)-2-azabicyclo[3.1.0]hex-2-
yl)carbony1)-2-methylpropyl)carbamate
D
methyl ((1S)-1-(((1R,3S,5R)-3-(7-((2-((3S)-
2-((2S)-2-((methoxycarbonyl)amino)-3-
methylbutanoy1)-2-azabicyclo[3.1.0]hex-3-
D-10 y1)-1H-naphtho[1,2-d]imidazol-7-
yl)ethyny1)-1H-naphtho[1,2-d]imidazol-2-
y1)-2-azabicyclo[3.1.0]hex-2-yl)carbony1)-2-
methylpropyl)carbamate
D
methyl ((1S)-2-((1R,3S,5R)-3-(7-((2-
((1R,3S,5R)-242S)-2-
((methoxycarbonyl)amino)-3-
methylbutanoy1)-2-azabicyclo[3.1.0]hex-3-
D-11
y1)-1H-naphtho[1,2-d]imidazol-7-
y1)ethynyl)-1H-naphtho[1,2-d]imidazol-2-
y1)-2-azabicyclo[3.1.0]hex-2-y1)-2-oxo-1-
(tetrahydro-2H-pyran-4-y1)ethyl)carbamate
M1 D methyl ((1S)-1-(((2S)-2-(4-(4-((4-(2-((2S)-
1-
((2S)-2-((methoxycarbonyl)amino)-3-
methylbutanoy1)-2-pyrrolidiny1)-1H-
imidazol-4-yl)phenyl)ethynyl)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidinyl)carbony1)-2-
methylpropyl) carbamate
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Compound lb EC50 Range Name
No. (nM)
M2 0.24 C dimethyl (1,2 -ethynediylbi s (4,1-
phenylene-
1H-imidazole-4,2-diy1(2S)-2,1-
pyrrolidinediyl ((1R)-2-oxo-1-phenyl-2,1-
ethanediy1))) biscarbamate
M2.1 D dimethyl (1,2-ethynediylbis(4,1-phenylene-
1H-imidazole-4,2-diy1(2S)-2,1-
pyrrolidinediy1((1S)-2-oxo-1-(tetrahydro-2H-
pyran-4-y1)-2,1-ethanediy1)))biscarbamate
M3 D methyl ((1S)-1-(((1R,3S,5R)-3-(4-(4-((4-(2-
((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)
amino)-3-methylbutanoy1)-2-
azabicyclo [3.1.0] hex-3-y1)-1H-imidazol-4-
yl)phenyl)ethynyl) pheny1)-1H-imidazol-2-
y1)-2-azabicyclo[3.1.0] hex-2-yl)carbony1)-2-
methylpropyl)carbamate
M4 D dimethyl (1,2 -ethynediylbi s (4,1-
phenylene-
1H-imidazole-4,2-diy1(1R,3S,5R)-2-
azabicyclo [3.1.0]hexane-3,2-diy1((2S)-1-
oxo-1,2-butanediy1)))biscarbamate
M5 C dimethyl (1,2-ethynediylbis(4,1-phenylene-
1H-imidazole-4,2-diy1(1R,3S,5R)-2-
azabicyclo [3.1.0]hexane-3,2-diy1((lS)-1-
cyclobuty1-2-oxo-2,1-
ethanediy1)))biscarbamate
M6 0.0033 D dimethyl (1,2-ethynediylbis(4,1-
phenylene-
1H-imidazole-4,2-diy1(1R,3S,5R)-2-
azabicyclo [3.1.0]hexane-3,2-diy1((1S)-2-
oxo-1-(tetrahydro-2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
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Compound lb EC50 Range Name
No. (nM)
M7 D methyl ((1S)-1-(((1R,3S,5R)-3-(4-(4-((4-(2-
((1R,3S,5R)-2-((2S)-2-((methoxycarbonyl)
amino)-2-(tetrahydro-2H-pyran-4-yl)ac ety1)-
2-azabicyclo [3.1.0]hex-3 -y1)-1H-imidazol-4-
yl)phenyl)ethynyl)pheny1)-1H-imi dazol-2-
y1)-2-azab icycl o [3. 1.0]hex-2-yl)c arbony1)-2-
methylpropyl)c arb amate
M8 D methyl ((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-
(4-((4-(241R,3S,5R)-2-((2S)-2-
((methoxycarbonyl)amino)-3-
methylbutanoy1)-2-azabicyclo [3 .1.0]hex-3 -
y1)-1H-imidazol-4-
yl)phenyl)ethynyl)pheny1)-1H-imi dazol-2-
y1)-2-azab icycl o [3. 1.0]hex-2-yl)c arbony1)-2-
methylpropyl)c arb amate
M9 D methyl ((1S)-1-(((1R,3S,5R)-3-(4-chloro-5-
(4-((4-(4-chloro-2-((1R,3S,5R)-242S)-2-
((methoxycarbonyl)amino)-3-
methylbutanoy1)-2-azabicyclo [3 .1.0]hex-3 -
y1)-1H-imidazol-5-
yl)phenyl)ethynyl)pheny1)-1H-imi dazol-2-
y1)-2-azab icycl o [3. 1.0]hex-2-yl)c arbony1)-2-
methylpropyl)c arb amate
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Compound lb EC50 Range Name
No. (nM)
D methyl ((1S)-2-((1R,3S,5R)-3-(4-(4-((4-(4-
chloro-2-((lR,3S,5R)-242S)-2-
((methoxycarbonyl)amino)-2-(tetrahydro-
M9.1
2H-pyran-4-yl)acety1)-2-
azabicyclo[3.1.0]hex-3-y1)-1H-imidazol-5-
yl)phenyl)ethynyl)pheny1)-1H-imidazol-2-
y1)-2-azabicyclo[3.1.0]hex-2-y1)-2-oxo-1-
(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
D
dimethyl (1,2-ethynediylbis(4,1-
phenylene(4-chloro-1H-imidazole-5,2-
M9.2 diy1)(1R,3S,5R)-2-azabicyclo[3.1.0]hexane-
3,2-diy1((lS)-2-oxo-1-(tetrahydro-2H-pyran-
4-y1)-2,1-ethanediy1)))biscarbamate
D methyl ((1S)-1-(((1R,3S,5R)-3-(4-(4-
((1S,2S)-2-(4-(2-((1R,3S,5R)-242S)-2-
((methoxycarbonyl)amino)-3-
M9 methylbutanoy1)-2-azabicyclo[3.1.0]hex-3-
.3
y1)-1H-imidazol-4-
y1)phenyl)cyclopropyl)phenyl)-1H-imidazol-
2-y1)-2-azabicyclo[3.1.0]hex-2-y1)carbonyl)-
2-methylpropyl)carbamate
D
dimethyl ((1S,2S)-1,2-
cyclopropanediylbis(4,1-phenylene-1H-
imidazole-4,2-diy1(1R,3S,5R)-2-
M9.4
azabicyclo[3.1.0]hexane-3,2-diy1(( 1 S)-2-
oxo-1-(tetrahydro-2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
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Compound lb EC50 Range Name
No. (nM)
D methyl ((1S)-1-(((1R,3 S,5R)-3 -(444-
((1R,2R)-2-(4-(2-((1R,3 S,5R)-2-((2S)-2-
((methoxycarbonyl)amino)-3-
M9 methylbutanoy1)-2-azabicyclo [3.1.0]hex-3 -
.5
y1)-1H-imidazol-4-
y1)phenyl)cyclopropyl)phenyl)-1H-imidazol-
2-y1)-2-azabicyclo[3.1.0]hex-2-y1)carbonyl)-
2-methylpropyl)carbamate
D
dimethyl ((1R,2R)-1,2-
cyclopropanediylbis (4,1-phenylene-1H-
imidazole-4,2-diy1(1R,3S,5R)-2-
M9.6
azabicyclo[3.1.0]hexane-3,2-diy1(( 1 S)-2-
oxo-1-(tetrahydro-2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
methyl ((1S)-1-(((2S,4S)-2-(4-(4-((lS,2S)-2-
(4-(2-((2S,4S)-142S)-2-
((methoxycarbonyl)amino)-3-
M9.7 D
methylbutanoy1)-4-methyl-2-pyrrolidiny1)-
1H-imidazol-4-
yl)phenyl)cyclopropyl)pheny1)-1H-imidazol-
2-y1)-4-methyl-l-pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
M9.8 D dimethyl ((lS,2S)-1,2-
cyclopropanediylbis(4,1-phenylene-1H-
imidazole-4,2-diy1((2S,4S)-4-methy1-2,1-
pyrrolidinediy1)((lS)-2-oxo-1-(tetrahydro-
2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
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Compound lb EC50 Range Name
No. (nM)
M10 D methyl ((1S)-1-(((2S,5S)-2-(4-(4-((4-(2-
((2S,5S)-142S)-2-
((methoxycarbonyl)amino)-3-
methylbutanoy1)-5-methyl-2-pyrrolidiny1)-
1H-imidazol-4-yl)phenyl)ethynyl)pheny1)-
1H-imidazol-2-y1)-5-methyl-1-pyrrolidinyl)
carbonyl)-2-methylpropyl)carbamate
Mll D dimethyl (1,2-ethynediylbis(4,1-phenylene-
1H-imidazole-4,2-diy1((2S,5S)-5-methy1-2,1-
pyrrolidinediy1)((2S)-1-oxo-1,2-butanediy1)))
biscarbamate
M12 D dimethyl (1,2-ethynediylbis(4,1-phenylene-
1H-imidazole-4,2-diy1((2S,5S)-5-methy1-2,1-
pyrrolidinediy1)((1S)-2-oxo-1-(tetrahydro-
2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
D
methyl ((1S)-1-(((2S,5S)-2-(4-(4-((4-(2-
((2S,5 S)-1-((2 S)-2-
((methoxycarbonyl)amino)-2-(tetrahydro-
M12.1
2H-pyran-4-yl)acety1)-5-methyl-2-
pyrrolidiny1)-1H-imidazol-4-
y1)phenyl)ethynyl)pheny1)-1H-imidazol-2-
y1)-5-methyl-1-pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
D
methyl ((1S)-1-(((2S,4S)-2-(4-(4-((4-(2-
((2S,4S)-1-((2 S)-2-
((methoxycarbonyl)amino)-3-
M12.2
methylbutanoy1)-4-methyl-2-pyrrolidiny1)-
1H-imidazol-4-yl)phenyl)ethynyl)pheny1)-
1H-imidazol-2-y1)-4-methyl-1-
pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
D
dimethyl (1,2-ethynediylbis(4,1-phenylene-
1H-imidazole-4,2-diy1((2S,4S)-4-methy1-2,1-
M12.3 0.014 pyrrolidinediy1)((lS)-2-oxo-1-(tetrahydro-
2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
D
methyl ((1S)-1-(((2S,4S)-2-(4-(4-((4-(4-
chloro-242S,4S)-142S)-2-
((methoxycarbonyl)amino)-3-
M12 methylbutanoy1)-4-methyl-2-pyrrolidiny1)-
.4
1H-imidazol-5-yl)phenyl)ethynyl)pheny1)-
1H-imidazol-2-y1)-4-methyl-1-
pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
D
methyl ((1S)-1-(((2S,4S)-2-(4-chloro-5-(4-
((4-(4-chloro-242S,4S)-142S)-2-
((methoxycarbonyl)amino)-3-
M12 methylbutanoy1)-4-methyl-2-pyrrolidiny1)-
.5
1H-imidazol-5-yl)phenyl)ethynyl)pheny1)-
1H-imidazol-2-y1)-4-methyl-1-
pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
D methyl ((1 S)-2-((2 S,4 S)-2-(4-(444-(4-
chloro-242 S,4S)-1-((2S)-2-
((methoxyc arb onyl)amino)-2-(tetrahydro-
M12.6
2H-pyran-4-yl)ac ety1)-4-methy1-2-
pyrrolidiny1)-1H-imidazol-5-
y1)phenyl)ethynyl)pheny1)-1H-imidazol-2-
y1)-4-methyl-1-pyrrolidiny1)-2-oxo-1-
(tetrahydro-2H-pyran-4-yl)ethyl)carbamate
D
dimethyl (1,2-ethynediylbis (4,1-
phenylene(4-chloro-1H-imidazole-5,2-
diy1)((2 S,4 S)-4-methy1-2,1 -
M12.7 0.061
pyrrolidinediy1)((lS)-2-oxo-1-(tetrahydro-
2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
D
methyl ((1 S)-2-((2 S,4 S)-2-(4-(4-((4-(2-
((2S,4S)-1-((2S)-2-
((methoxycarbonyl)amino)-3-
M12.8
methylbutanoy1)-4-methyl-2-pyrrolidiny1)-
1H-imidazol-4-yl)phenyl)ethynyl)pheny1)-
1H-imidazol-2-y1)-4-methyl-1-pyrrolidiny1)-
2-oxo-1-(tetrahydro-2H-pyran-4-
yl)ethyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
D methyl ((1S)-1-(((1S,3S,5S)-3-(4-(4-((4-(2-
((1S,3S,5S)-242S)-2-
((methoxycarbonyl)amino)-3-
methylbutanoy1)-5-methyl-2-
M12.9
azabicyclo[3.1.0]hex-3-y1)-1H-imidazol-4-
yl)phenyl)ethynyl)pheny1)-1H-imidazol-2-
y1)-5-methy1-2-azabicyclo[3.1.0]hex-2-
yl)carbony1)-2-methylpropyl)carbamate
M13 D methyl ((1S)-1-(((1R,3S,5R)-3-(4-(4-((E)-2-
(4-(2-((1R,3S,5R)-2-((2S)-2-
((methoxycarbonyl) amino)-3-
methylbutanoy1)-2-azabicyclo[3.1.0] hex-3-
y1)-1H-imidazol-4-y1)phenyl)vinyl) pheny1)-
1H-imidazol-2-y1)-2-azabicyclo[3.1.0] hex-
2-yl)carbony1)-2-methylpropyl)carbamate
M14 0.017 D dimethyl ((E)-1,2-ethenediylbis(4,1-
phenylene-1H-imidazole-4,2-
diy1(1R,3S,5R)-2-azabicyclo [3.1.0]hexane-
3,2-diy1((1S)-2-oxo-1-(tetrahydro-2H-pyran-
4-y1)-2,1-ethanediy1))) biscarbamate
M15 D methyl ((1S)-241R,3S,5R)-3-(4-(4-((E)-2-
(4-(2-((1R,3S,5R)-2-((2S)-2-
((methoxycarbonyl) amino)-3-
methylbutanoy1)-2-azabicyclo[3.1.0] hex-3-
y1)-1H-imidazol-4-y1)phenyl)vinyl) pheny1)-
1H-imidazol-2-y1)-2-azabicyclo[3.1.0] hex-
2-y1)-2-oxo-1-(tetrahydro-2H-pyran-4-
yl)ethyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
D methyl ((1S)-14(2S)-2-(4-(4-((4-(242S)-
4,4-difluoro-142 S)-2-
((methoxycarbonyl)amino)-3-
N1
methylbutanoy1)-2-pyrrolidiny1)-1H-
imidazol-4-yl)phenyl)ethynyl)pheny1)-1H-
imidazol-2-y1)-4,4-difluoro-1-
pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
D
dimethyl (ethyne-1,2-diylbis(4,1-phenylene-
1H-imidazole-5,2-diy1((2S)-4,4-
N2 difluoropyrrolidine-2,1-diy1)((1S)-2-oxo-1-
(tetrahydro-2H-pyran-4-yl)ethane-2,1-
diy1)))biscarbamate
A methyl ((1R)-242S)-2-(5-(444-(242S)-
4,4-difluoro-142 S)-2-
((methoxycarbonyl)amino)-2-(tetrahydro-
N3 0.25
2H-pyran-4-yl)acetyl)pyrrolidin-2-y1)-1H-
imidazol-5-yl)phenyl)ethynyl)pheny1)-1H-
imidazol-2-y1)-4,4-difluoropyrrolidin-1-y1)-
2-oxo-1-(tetrahydro-2H-pyran-4-
y1)ethyl)carbamate
A
dimethyl (ethyne-1,2-diylbis(4,1-phenylene-
1H-imidazole-5,2-diy1((2S)-4,4-
N4 0.18 difluoropyrrolidine-2,1-diy1)((1R)-2-oxo-1-
(tetrahydro-2H-pyran-4-yl)ethane-2,1-
diy1)))biscarbamate
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Compound lb EC50 Range Name
No. (nM)
D methyl ((1S)-1-(((3S)-3-(4-(4-((4-(2-((3 S)-
4-
((2S)-2-((methoxycarb onyl)amino)-3-
methylbutanoy1)-3 -morpholiny1)-1H-
N5
imidazol-4-yl)phenyl)ethynyl)pheny1)-1H-
imidazol-2-y1)-4-morpholinyl)carbony1)-2-
methylpropyl)carbamate
A methyl ((1S)-1-(((2S)-2-(4-(4-((4-(2-((2 S)-
1-
((2S)-2-((methoxycarb onyl)amino)-3-
N6
methylbutanoy1)-2-piperidiny1)-1H-imidazol-
4-yl)phenyl)ethynyl)pheny1)-1H-imidazol-2-
y1)-1-piperidinyl)carbony1)-2-
methylpropyl)carbamate
D
methyl ((1S)-1-(((2S,4S)-4-hydroxy-2-(4-(4-
((4-(2-((2S,4 S)-4-hydroxy-1-((2S)-2-
((methoxycarb onyl)amino)-3-
N7 methylbutanoy1)-2-pyrrolidiny1)-1H-
imidazol-4-yl)phenyl)ethynyl)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
methyl ((1S)-1-(((2S)-2-(4-(4-((4-(2-((2S)-1-
((2S)-2-((methoxycarbonyl)amino)-3-
methylbutanoy1)-4-methylene-2-
N7.1 pyrrolidiny1)-1H-imidazol-4-
yl)phenyl)ethynyl)pheny1)-1H-imidazol-2-
y1)-4-methylene-l-pyrrolidinyl)carb ony1)-2-
methylpropyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
methyl ((1S)-1-(((2S,4R)-4-hydroxy-2-(4-(4-
((4-(2-((2 S,4R)-4-hydroxy-1-((2S)-2-
((methoxycarbonyl)amino)-3-
N7.2 methylbutanoy1)-2-pyrrolidiny1)-1H-
imidazol-4-yl)phenyl)ethynyl)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
A methyl((lS)-1-(((lS,3S,5S)-3-(4-(4-((4-(2-
((1S,3S,5S)-2-((2S)-2-
((methoxycarbonyl)amino)-3-
N8 methylbutanoy1)-2-azabicyclo [3.1.0]hex-3 -
y1)-1H-imidazol-4-
y1)phenyl)ethynyl)phenyl)-1H-imidazol-2-
y1)-2-azabicyclo[3.1.0]hex-2-y1)carbonyl)-2-
methylpropyl)carbamate
D
dimethyl (1,2-ethynediylbis(4,1-phenylene-
1H-imidazole-4,2-diy1(1S,3S,5 S)-2-
N9 azabicyclo[3.1.0]hexane-3,2-diy1(( 1 S)-2-
oxo-1-(tetrahydro-2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
D methyl ((1S)-1-(((2S)-2-(4-(3-((3-(2-((2 S)-
1-
((2S)-2-((methoxycarbonyl)amino)-3-
J1
methylbutanoy1)-2-pyrrolidiny1)-1H-
imidazol-4-yl)phenyl)ethynyl)pheny1)-1H-
imidazol-2-y1)-1-pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
A methyl ((1 S)-2-methyl- -(((2 S)-2-(4-(3-((3
-
(2-((2S)-2-pyrrolidiny1)-1H-imidazol-4-
J1.1 338.8 yl)phenyl)ethynyl)pheny1)-1H-imidazol-2-
y1)-1-
pyrrolidinyl)c arb onyl)propyl)c arb amate
C
dimethyl (1,2-ethynediylbis(3,1-phenylene-
1H-imidazole-4,2-diy1(2 S)-2,1-
J2 0.12
pyrrolidinediy1((lS)-2-oxo-1-(tetrahydro-2H-
pyran-4-y1)-2,1-ethanediy1)))biscarbamate
D
methyl ((1S)-1-(((1R,3 S,5R)-3 -(4-(3 -((3 -(2-
(( 1 R,3 S,5R)-242 S)-2-
((methoxyc arb onyl)amino)-3-
methylbutanoy1)-2-azabicyclo [3.1.0]hex-3 -
J3
y1)-1H-imidazol-4-
y1)phenyl)ethynyl)phenyl)-1H-imidazol-2-
y1)-2-azabicyclo[3.1.0]hex-2-y1)carbonyl)-2-
methylpropyl)carbamate
D
dimethyl (1,2-ethynediylbis(3,1-phenylene-
1H-imidazole-4,2-diy1(1R,3 S,5R)-2-
J4 azabicyclo[3.1.0]hexane-3,2-diy1(( 1 S)-2-
oxo-1-(tetrahydro-2H-pyran-4-y1)-2,1-
ethanediy1)))biscarbamate
D methyl ((1 S)-1-(((2S)-2-(4-(4'-((4-(2-((2S)-
1-
((2S)-2-((methoxyc arb onyl)amino)-3-
J5 methylbutanoy1)-2-pyrrolidiny1)-1H-
imidazol-4-yl)phenyl)ethyny1)-4-
biphenyly1)-1H-imidazol-2-y1)-1-
pyrrolidinyl)carbony1)-2-
methylpropyl)carbamate
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Compound lb EC50 Range Name
No. (nM)
N7.3 D
N7.8 D
N7.9 1.10 B
N10 D
N11 D
N11.1 0.00147 D
N11.2 D
N12 D
N13 D
N14 D
N15 D
N16.1 C
N16.2 D
N16.3 B
N17.1 D
N17.2 0.13 C
N22 D
N23 D
N24 C
N25
The compounds of the present disclosure may inhibit HCV by mechanisms in
addition to or other than NS5A inhibition. In one embodiment the compounds of
the
present disclosure inhibit HCV replicon and in another embodiment the
compounds
of the present disclosure inhibit NS5A.
It will be evident to one skilled in the art that the present disclosure is
not
limited to the foregoing illustrative examples, and that it can be embodied in
other
specific forms without departing from the essential attributes thereof It is
therefore
desired that the examples be considered in all respects as illustrative and
not
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restrictive, reference being made to the appended claims, rather than to the
foregoing
examples, and all changes which come within the meaning and range of
equivalency
of the claims are therefore intended to be embraced therein.
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