U.S. patent application number 15/019599 was filed with the patent office on 2017-01-12 for c-3 novel triterpenone with c-28 reverse amide derivatives as hiv inhibitors.
This patent application is currently assigned to HETERO RESEARCH FOUNDATION. The applicant listed for this patent is HETERO RESEARCH FOUNDATION. Invention is credited to Gazula Levi David Krupadanam, Adulla Panduranga Reddy, Bandi Parthasaradhi Reddy, Kasireddy Bhaskar Reddy, Kura Rathnakar Reddy, Lanka VL Subrahmanyam.
Application Number | 20170008921 15/019599 |
Document ID | / |
Family ID | 57153504 |
Filed Date | 2017-01-12 |
United States Patent
Application |
20170008921 |
Kind Code |
A1 |
Reddy; Bandi Parthasaradhi ;
et al. |
January 12, 2017 |
C-3 NOVEL TRITERPENONE WITH C-28 REVERSE AMIDE DERIVATIVES AS HIV
INHIBITORS
Abstract
The invention relates to C-3 novel triterpenone with C-28
reverse amide derivatives, related compounds, and pharmaceutical
compositions useful for the therapeutic treatment of viral diseases
and particularly HIV mediated diseases. ##STR00001##
Inventors: |
Reddy; Bandi Parthasaradhi;
(Telangana, IN) ; Reddy; Kura Rathnakar;
(Telangana, IN) ; Krupadanam; Gazula Levi David;
(Telangana, IN) ; Reddy; Adulla Panduranga;
(Telangana, IN) ; Reddy; Kasireddy Bhaskar;
(Telangana, IN) ; Subrahmanyam; Lanka VL;
(Telangana, IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
HETERO RESEARCH FOUNDATION |
Telangana |
|
IN |
|
|
Assignee: |
HETERO RESEARCH FOUNDATION
Telangana
IN
|
Family ID: |
57153504 |
Appl. No.: |
15/019599 |
Filed: |
February 9, 2016 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 31/18 20180101;
C07J 63/008 20130101 |
International
Class: |
C07J 63/00 20060101
C07J063/00 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 9, 2015 |
IN |
623/CHE/2015 |
Claims
1. A compound of the formula (1): ##STR00167## wherein, R.sub.1 is
substituted or unsubstituted C.sub.1-C.sub.6 alkyl, ##STR00168##
(wherein R.sub.a is selected from hydrogen, substituted or
unsubstituted C.sub.1-C.sub.6 alkyl, or substituted or
unsubstituted C.sub.3-C.sub.8 cycloalkyl); R.sub.2 is selected from
hydrogen, substituted or unsubstituted C.sub.1-C.sub.6 alkyl;
R.sub.3 and R.sub.4 are independently selected from substituted or
unsubstituted C.sub.1-C.sub.6 alkyl, substituted or unsubstituted
amine, substituted or unsubstituted C.sub.3-C.sub.8 cycloalkyl and
R.sub.3 and R.sub.4 are taken together with the carbon atom to
which they are attached to form substituted or unsubstituted
C.sub.3-C.sub.8 cycloalkyl, epoxide, oxetane or azetidine; R.sub.5
and R.sub.6 are independently selected from hydrogen, substituted
or unsubstituted C.sub.1-C.sub.6 alkyl and R.sub.5 and R.sub.6 are
taken together with the carbon atom to which they are attached form
substituted or unsubstituted C.sub.3-C.sub.8 cycloalkyl or R.sub.5
and R.sub.6 together represent oxo; R.sub.7 is selected from
hydrogen, substituted or unsubstituted C.sub.1-C.sub.6 alkyl,
substituted or unsubstituted C.sub.1-C.sub.6 alkoxy, substituted or
unsubstituted amino, substituted or unsubstituted C.sub.1-C.sub.6
amino alkyl, substituted or unsubstituted C.sub.3-C.sub.8
cycloalkyl, substituted or unsubstituted C.sub.6-C.sub.12 aryl,
substituted or unsubstituted heterocyclyl, substituted or
unsubstituted heteroaryl, or --S(O).sub.2R.sub.b; wherein the
substituents are independently selected from one or more R.sub.m;
R.sub.m is selected from halo, C.sub.1-C.sub.6 alkyl, haloalkyl,
amino, --C(O)OR.sub.c, substituted or unsubstituted heterocyclyl,
substituted or unsubstituted heterocyclylalkyl, substituted or
unsubstituted heteroaryl or --S(O).sub.2R.sub.b; R.sub.b and
R.sub.c are independently selected from substituted or
unsubstituted C.sub.1-C.sub.6 alkyl or substituted or unsubstituted
C.sub.6-C.sub.12 aryl; `n` is an integer selected from 0, 1 or 2;
pharmaceutically acceptable salts, pharmaceutically acceptable
solvates, pharmaceutically acceptable hydrates, tautomers,
stereoisomers, ester prodrugs, or combination thereof.
2. The compound of claim 1, wherein R.sub.1 is ##STR00169##
3. The compound according to claim 1, which is a compound of the
formula (1A): ##STR00170## wherein, R.sub.2, R.sub.5, R.sub.6,
R.sub.7 and `n` are same as defined in claim 1.
4. The compound according to claim 1, which is a compound of the
formula (1B): ##STR00171## wherein, R.sub.2, R.sub.5, R.sub.6,
R.sub.7 and `n` are same as defined in claim 1; X is selected from
--O--, --CH.sub.2O--, --CH.sub.2N--, or (--CH.sub.2--).sub.m; `m`
is an integer selected from 1, 2, 3 or 4.
5. A compound selected from the group consisting of:
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chlorobenzamido-
)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,-
5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)-
carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(1-(4-chlorophenyl-
)
cyclopropane-1-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,1-
1a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobut-
ane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-benzamido-2-methyl
propanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,11,
11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl-
)-2,2-dimethyl cyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(3,4-dichlorobenza-
mido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3-
a,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(pyrazine-2-carboxamido)propanamido)-2-oxo-3,-
3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(6-aminonicotinami-
do)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,-
4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(5-methylpyrazine-2-carboxamido)propanamido)--
2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-1-(tert-butox-
y
carbonyl)pyrrolidine-2-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,-
5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13-
,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl-
cyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(4-ethylpiperaz-
in-1-yl)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamet-
hyl-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-(piperidin-1-yl)acetamido)propanamido)-2-o-
xo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-amino-2-methyl
propanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-
-oxo-3,3a,4, 5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(1H-benzo[d]imidaz-
ole-5-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamet-
hyl-2-oxo-3,3a,4,
5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]-
chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-(6-methylpyridin-3-yl)-1H-benzo[d]imidazol-
e-5-carboxamido)propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b-
,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-di-
methylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2,4-dimethylthiaz-
ole-5-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamet-
hyl-2-oxo-3,3a,4,
5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]-
chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-(pyrazin-2-yl)-1H-benzo[d]imidazole-5-carb-
oxamido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,-
8,8,11a-pentamethyl-3a-(2-methyl-2-(1-methyl-1H-imidazole-2-carboxamido)pr-
opanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadec-
ahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane--
1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-3a-(2-(3-iso-
propyl-1H-pyrazole-5-carboxamido)-2-methylpropanamido)-5a,5b,8,8,11a-penta-
methyl-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-morpholinobenzamido)propanamido)-2-oxo-3,3-
a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(3,
5-dimethylisoxazole-4-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b-
,8,8,11a-pentamethyl-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(4-methyl-1H-imidazol-1-yl)benzamido)propa-
namido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-c-
arboxylic acid,
(1S,3R)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chlorobenzamido-
)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,-
5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)-
carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-fluorobenzamido-
)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,-
5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-methylbenzamido)propanamido)-2-oxo-3,3a,4,-
5,5a,5b,6,7,
7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl-
)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(furan-3-carboxami-
do)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,-
4,5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(trifluoromethyl)benzamido)propanamido)-2--
oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(furan-2-carboxami-
do)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,-
4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(1-phenylcyclopentane-1-carboxamido)propanami-
do)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro--
2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carbo-
xylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(quinoline-2-carboxamido)propanamido)-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(3-methylpicolinamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-methylfuran-3-carboxamido)propanamido)-2-o-
xo-3,3a,4, 5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-morpholinonicotinamido)propanamido)-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(pyrimidine-2-carboxamido)propanamido)-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2,5-dimethylfuran-
-3-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-
-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(1,1-dioxidothi-
o
morpholino)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pen-
tamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecah-
ydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1--
carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(piperidine-4-carboxamido)propanamido)-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-2-amino-3-met-
hyl
butanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-
-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(methylsulfonyl)benzamido)propanamido)-2-o-
xo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((S)-piperidine-3-carboxamido)propanamido)-2--
oxo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(4-chlorophenyl-
)
acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2--
oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)prop-
anamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecah-
ydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1--
carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-aminothiazole-4-
-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(5-methyl-1,3,4-oxadiazol-2-yl)benzamido)p-
ropanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octade-
cahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-
-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-(1,1-dioxidothi-
o
morpholino)benzamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pen-
tamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecah-
ydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1--
carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-((1,1-dioxidoth-
io
morpholino)methyl)benzamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8-
,11a-penta methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(dimethyl-
amino)
acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentameth-
yl-2-oxo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(6-methylpicolinamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b, 6,7,
7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl-
)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(6-methylnicotinamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b, 6,7,
7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl-
)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-pivalamidopropanamido)-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,11,
11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl-
)-2,2-dimethyl cyclobutane-1-carboxylic acid, sodium
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(methylsulfonamido)propanamido)-2-oxo-3,3a,4,-
5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]-
chrysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylate,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((ethoxycarbonyl)a-
mino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3-
a,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((4-chlorophenyl)
sulfonamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(cyclohexanecarbox
amido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((pyridin-2-ylmethyl)amino)propanamido)-2-oxo-
-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((4-chlorobenzyl)a-
mino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3-
a,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(((1-(4-chlorophen-
yl)
cyclopropyl)methyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,1-
1a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobut-
ane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(5-chloropicolinam-
ido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a-
,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-3-yl)ureido)propanamido)--
2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-2-yl)ureido)propanamido)--
2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((2-(dimethylamino-
)
ethyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl--
2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylprop-
an
amido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7-
a,8,9,10,11,11a,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,-
2-dimethyl cyclobutane-1-carboxylic acid hydrochloride,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-butoxycarbo-
nyl)
amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-o-
xo-3,3a,4,5,5a,5b, 6,7,
7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl-
)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid, and
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((S)-pyrrolidine-2-carboxamido)propanamido)-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride, or pharmaceutically acceptable salts, solvates,
hydrates and prodrugs thereof.
6. A compound selected from the group consisting of:
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclobutane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclobutane-1-carboxamido)-2-oxo-
-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclohexane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclopentane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclohexane-1-carboxamido)-2-oxo-
-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclopentane-1-carboxamido)-2-ox-
o-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(4-methylbenzamido)cyclohexane-1-carboxamido)-2-oxo-3,-
3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclopropane-1-carboxamido)-2-ox-
o-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-fluorobenzamido-
)
cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(4-methylbenzamido)cyclopropane-1-carboxamido)-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-2-oxo-3a-(1-(pyrimidine-2-carboxamido)cyclobutane-1-carboxam-
ido)-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid,
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(2-morpholinonicotinamido)cyclobutane-1-carboxamido)-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid, and
(1S,3R)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid, or
pharmaceutically acceptable salts, solvates, hydrates and prodrugs
thereof.
7. A pharmaceutical composition comprising a compound according to
claim 1; and at least one pharmaceutically acceptable
excipient.
8. The pharmaceutical composition according to claim 7, wherein the
pharmaceutically acceptable excipient is a carrier or diluent.
9. A method for preventing, ameliorating or treating a viral
mediated disease, disorder or syndrome in a subject in need thereof
comprising administering to the subject a therapeutically effective
amount of a compound according to claim 1.
10. The method according to claim 9, wherein the viral mediated
disease, disorder or syndrome is HIV infection, HBV infection, HCV
infection, a retroviral infection genetically related to AIDS,
respiratory disorders (including adult respiratory distress
syndrome (ARDS)), inflammatory disease, or a combination
thereof.
11. A method of treating HIV in a subject in need thereof
comprising administering to the subject a therapeutically effective
amount of a compound according to claim 1.
12. A method for preventing, ameliorating or treating a viral
mediated disease, disorder or syndrome in a subject in need thereof
comprising administering to the subject the pharmaceutical
composition according to claim 11 comprising a therapeutically
effective amount of the compound.
13. The method according to claim 12, wherein the viral mediated
disease, disorder or syndrome is HIV infection, HBV infection, HCV
infection, a retroviral infection genetically related to AIDS,
respiratory disorders (including adult respiratory distress
syndrome (ARDS)), inflammatory disease, or a combination
thereof.
14. A pharmaceutical composition comprising a compound according to
claim 5 and at least one pharmaceutically acceptable excipient.
15. A method for preventing, ameliorating or treating a viral
mediated disease, disorder or syndrome in a subject in need thereof
comprising administering to the subject a therapeutically effective
amount of a compound according to claim 5.
16. A method of treating HIV in a subject in need thereof
comprising administering to the subject a therapeutically effective
amount of a compound according to claim 5.
17. A pharmaceutical composition comprising a compound according to
claim 6 and at least one pharmaceutically acceptable excipient.
18. A method for preventing, ameliorating or treating a viral
mediated disease, disorder or syndrome in a subject in need thereof
comprising administering to the subject a therapeutically effective
amount of a compound according to claim 6.
19. A method of treating HIV in a subject in need thereof
comprising administering to the subject a therapeutically effective
amount of a compound according to claim 6.
Description
[0001] This application claims the benefit of Indian provisional
application no 623/CHE/2015 filed on 9 Feb. 2015 which is hereby
incorporated by reference in its entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to C-3 novel triterpenone with
C-28 reverse amide derivatives and related compounds, compositions
useful for therapeutic treatment of viral diseases and particularly
HIV mediated diseases.
BACKGROUND OF THE INVENTION
[0003] The Human Immunodeficiency Virus (HIV) has now been
established as the causative agent of the Acquired Immunodeficiency
Syndrome (AIDS) for over 20 years (Science 1983, 220, 868-871; N.
Eng. J. Med. 1984, 311, 1292-1297). AIDS is characterized by the
destruction of the immune system, particularly of CD4+T-cells. HIV
is a retrovirus, and the HIV life cycle encompasses several crucial
steps, starting from the attachment of the virus to the host cell
membrane and finishing with the release of progeny virons from the
cell.
[0004] The natural compound betulinic acid, isolated from Syzygium
clavifolium and several other plant species was found to possess
anti-HIV activity. Chemical modifications were undertaken by
several research groups in an attempt to identify potent anti-HIV
agents by making semi-synthetic analogs of betulinic acid, leading
to the discovery of bevirimat as a compound with a novel mechanism
of action (J. Nat. Prod. 199457(2):243-7; J. Med. Chem. 1996,
39(5), 1016). Further studies shown that bevirimat acts by
disrupting Gag processing (Proc. Natl. Acad. Sci. USA 2003,
100(23):13555-60; Antimicrob. Agents. Chemother. 2001, 45(4),
1225-30; J. Virol. 2004, 78(2): 922-9; J. Biol. Chem. 2005,
280(51):42149-55; J. Virol. 2006, 80(12): 5716-22) and to be a
first-in-class maturation inhibitor with a potent activity against
HIV-1. Bevirimat went up to phase 2 clinical trials, in clinic
despite optimal plasma concentrations, not all patients given
bevirimat have a robust viral load reduction. It was reported that
non-respondant patients had more frequent base line Gag
polymorphisms near the capsid SP-1 cleavage site than responders.
(HIV gag polymorphism determines treatment response to bevirimat.
XVII international HIV drug resistance work shop Jun. 10-14, 2008,
Sitges, Spain).
[0005] Encouraged by these developments, medicinal chemists started
exploring betulinic acid derivatives and related compounds
intensively for their therapeutic activities. For example, WO
2014/093941 describes pharmaceutical compositions of betulin
derivatives; WO 2009/082819 describes preparation of 17-amino
lupane derivatives as anti-HIV agents; WO 2013/117137 describes
lupane triterpenoids derivatives and pharmaceutical use thereof; WO
2013/020245 describes carbonyl derivatives of betulin; WO
2009/082818 describes preparation of C21-keto lupane derivatives
for the treatment of HIV infections; WO 2011/100308 describes
preparation of betulin derivatives for treatment of HIV-1; WO
2013/090664 describes preparation of betulin derivatives for the
treatment of HIV; WO 2013/091144 describes preparation of
propenoate derivatives of betulin useful for the treatment of HIV;
WO 2013/090683 describes preparation of betulin propenoate
derivatives for the treatment of HIV.
[0006] Given the fact of the world wide epidemic level of AIDS,
there is a strong continued need for new effective drugs for
treatment of HIV infected patients, disease conditions and/or
disorders mediated by HIV by discovering new compounds with novel
structures and/or mechanism of action(s).
SUMMARY OF THE INVENTION
[0007] The present invention relates to the compounds of the
formula (1)
##STR00002##
wherein,
[0008] R.sub.1 can be substituted or unsubstituted C.sub.1-C.sub.6
alkyl,
##STR00003##
(wherein R.sub.a can be hydrogen, substituted or unsubstituted
C.sub.1-C.sub.6 alkyl, or substituted or unsubstituted
C.sub.3-C.sub.8 cycloalkyl);
[0009] R.sub.2 can be hydrogen, substituted or unsubstituted
C.sub.1-C.sub.6 alkyl, substituted or unsubstituted C.sub.2-C.sub.6
alkoxylalkoxy or substituted or unsubstituted C.sub.1-C.sub.6 amino
alkyl;
[0010] R.sub.3 and R.sub.4 can be independently selected from
substituted or unsubstituted C.sub.1-C.sub.6 alkyl, substituted or
unsubstituted amine, substituted or unsubstituted C.sub.3-C.sub.8
cycloalkyl, substituted or unsubstituted heterocyclyl or
substituted or unsubstituted heteroaryl or R.sub.3 and R.sub.4 can
be taken together with the carbon atom to which they are attached
to form substituted or unsubstituted C.sub.3-C.sub.8 cycloalkyl,
epoxide, oxetane or azetidine;
[0011] R.sub.5 and R.sub.6 can be independently selected from
hydrogen, substituted or unsubstituted C.sub.1-C.sub.6 alkyl and
R.sub.5 and R.sub.6 can be taken together with the carbon atom to
which they are attached to form substituted or unsubstituted
C.sub.3-C.sub.8 cycloalkyl or R.sub.5 and R.sub.6 can be together
represent oxo;
[0012] R.sub.7 can be hydrogen, substituted or unsubstituted
C.sub.1-C.sub.6 alkyl, substituted or unsubstituted C.sub.1-C.sub.6
alkoxy, substituted or unsubstituted amino, substituted or
unsubstituted C.sub.1-C.sub.6 amino alkyl, substituted or
unsubstituted C.sub.3-C.sub.8 cycloalkyl, substituted or
unsubstituted C.sub.6-C.sub.12 aryl, substituted or unsubstituted
heterocyclyl, substituted or unsubstituted heteroaryl, or
--S(O).sub.2R.sub.b; wherein the substituents can be independently
selected from one or more R.sub.m;
[0013] R.sub.m can be halo, C.sub.1-C.sub.6 alkyl, haloalkyl,
amino, --C(O)OR.sub.c, substituted or unsubstituted heterocyclyl,
substituted or unsubstituted heterocyclylalkyl, substituted or
unsubstituted heteroaryl or --S(O).sub.2R.sub.b;
[0014] R.sub.b and R.sub.c can be independently selected from
substituted or unsubstituted C.sub.1-C.sub.6 alkyl or substituted
or unsubstituted C.sub.6-C.sub.12 aryl;
[0015] `n` can be an integer selected from 0, 1 or 2;
[0016] pharmaceutically acceptable salts, pharmaceutically
acceptable solvates, pharmaceutically acceptable hydrates,
tautomers, stereoisomers, ester prodrugs, or combination
thereof.
[0017] It should be understood that the formula (1) structurally
encompasses all stereoisomers, including enantiomers,
diastereomers, racemates, and combinations thereof which may be
contemplated from the chemical structure of the genus described
herein.
[0018] It should be understood that the formula (1) structurally
encompasses all tautomers.
[0019] Also contemplated are prodrugs of the compounds of the
formula (1), including ester prodrugs.
[0020] According to one embodiment, there is provided a compound of
formula (1), wherein R.sub.1 is
##STR00004##
[0021] According to another embodiment, there is provided a
compound of formula (1), wherein R.sub.2 is hydrogen.
[0022] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.3 and R.sub.4 are
methyl.
[0023] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.3 and R.sub.4 together with
the carbon atom to which they are attached to form substituted or
unsubstituted C.sub.3-6 cycloalkyl.
[0024] According to yet another embodiment there is provided a
compound of formula (1), wherein the above said C.sub.3-6
cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.
[0025] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.5 and R.sub.6 are
hydrogen.
[0026] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.5 and R.sub.6 are
methyl.
[0027] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.5 and R.sub.6 together with
the carbon atom to which they are attached form C.sub.3-6
cycloalkyl.
[0028] According to yet another embodiment there is provided a
compound of formula (1), wherein the above said C.sub.3-6
cycloalkyl is cyclopropyl and cyclopentyl.
[0029] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.5 and R.sub.6 together
represent oxo.
[0030] According to yet another embodiment, there is provided a
compound of formula (1), wherein R.sub.7 is substituted phenyl;
wherein the substituents are chloro, methyl, fluoro,
trifluoromethyl, morpholine, methylsulfonyl, 4-Methyl-1H-imidazole,
oxazole, 1,3,4-Oxadiazole, thiomorpholine 1,1-dioxide or
4-methylthiomorpholine 1,1-dioxide.
[0031] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is substituted pyridine;
wherein the substituents are methyl, amino, morpholine or
chloro.
[0032] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is pyrrolidine.
[0033] According to yet another embodiment, there is provided a
compound of formula (1), wherein R.sub.7 is pyrrolidine which is
substituted with --C(O)OR.sub.c; wherein R.sub.c is tertiary
butyl.
[0034] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is pyrazine.
[0035] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is pyrazine which is
substituted with methyl.
[0036] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is furan which is
substituted with methyl.
[0037] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is piperazine which is
substituted with ethyl.
[0038] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is quinoline.
[0039] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is piperidine.
[0040] According to yet another embodiment, there is provided a
compound of formula (1), wherein R.sub.7 is thiomorpholine
1,1-dioxide.
[0041] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is
1H-benzo[d]imidazole.
[0042] According to yet another embodiment, there is provided a
compound of formula (1), wherein R.sub.7 is 1H-benzo[d]imidazole
which is substituted with methyl pyridine.
[0043] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is 1H-benzo[d]imidazole
which is substituted with pyrazine.
[0044] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is thiazole which is
substituted with methyl or amino.
[0045] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is imidazole which is
substituted with methyl.
[0046] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is pyrazole which is
substituted with isopropyl.
[0047] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is isoxazole which is
substituted with methyl.
[0048] According to yet another embodiment, there is provided a
compound of formula (1), wherein R.sub.7 is 1,3,4-Oxadiazole which
is substituted with methyl.
[0049] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is pyrimidine.
[0050] According to yet another embodiment, there is provided a
compound of formula (1), wherein R.sub.7 is
pyrazolo[1,5-a]pyrimidine.
[0051] According to yet another embodiment, there is provided a
compound of formula (1), wherein R.sub.7 is --S(O).sub.2R.sub.b;
wherein R.sub.b is methyl or 4-chloro phenyl.
[0052] According to yet another embodiment, there is provided a
compound of formula (1), wherein R.sub.7 is N,N-dimethyl amino.
[0053] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is tertiary butyl.
[0054] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is tertiary butoxy.
[0055] According to yet another embodiment there is provided a
compound of formula (1), wherein R.sub.7 is cyclohexyl.
[0056] According to yet another embodiment, there is provided a
compound of formula (1), wherein `n` is 0.
[0057] According to yet another embodiment, there is provided a
compound of formula (1), wherein `n` is 1.
[0058] According to yet another embodiment, there is provided a
compound of formula (1), wherein `n` is 2.
[0059] Accordingly, one other aspect of the present invention
provides compounds of formula (1A):
##STR00005##
wherein, [0060] R.sub.2, R.sub.5, R.sub.6, R.sub.7 and `n` are same
as defined in claim 1;
[0061] pharmaceutically acceptable salts, pharmaceutically
acceptable solvates, pharmaceutically acceptable hydrates,
tautomers, stereoisomers, ester prodrugs, or combination
thereof.
[0062] It should be understood that the formula (1A) structurally
encompasses all stereoisomers, including enantiomers,
diastereomers, racemates, and combinations thereof which may be
contemplated from the chemical structure of the genus described
herein.
[0063] It should be understood that the formula (1A) structurally
encompasses all tautomers.
[0064] Also contemplated are prodrugs of the compounds of the
formula (1A), including ester prodrugs.
[0065] According to one embodiment, there is provided a compound of
formula (1A), wherein R.sub.2 is hydrogen.
[0066] According to one embodiment, there is provided a compound of
formula (1A), wherein R.sub.5 and R.sub.6 are hydrogen.
[0067] According to one embodiment, there is provided a compound of
formula (1A), wherein R.sub.5 and R.sub.6 are methyl.
[0068] According to one embodiment, there is provided a compound of
formula (1A), wherein R.sub.5 and R.sub.6 together represent
oxo.
[0069] According to one embodiment, there is provided a compound of
formula (1A), wherein R.sub.5 and R.sub.6 together with the carbon
atom to which they are attached to form C.sub.3-6 cycloalkyl.
[0070] According to one embodiment, there is provided a compound of
formula (1A), wherein the above said C.sub.3-6 cycloalkyl is
cyclopropyl, cyclopentyl or cyclohexyl.
[0071] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is substituted phenyl;
wherein the substituents are chloro, methyl, fluoro,
trifluoromethyl, morpholine, methylsulfonyl, 4-Methyl-1H-imidazole,
oxazole, 1,3,4-Oxadiazole, thiomorpholine 1,1-dioxide or
4-methylthiomorpholine 1,1-dioxide.
[0072] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is substituted pyridine,
wherein the substituents are methyl, amino, morpholine or
chloro.
[0073] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is pyrrolidine.
[0074] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is pyrrolidine which is
substituted with --C(O)OR.sub.c; wherein R.sub.c is tertiary
butyl.
[0075] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is pyrazine.
[0076] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is pyrazine which is
substituted with methyl.
[0077] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is furan which is
substituted with methyl.
[0078] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is piperazine which is
substituted with ethyl.
[0079] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is quinoline.
[0080] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is piperidine.
[0081] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is thiomorpholine
1,1-dioxide.
[0082] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is
1H-benzo[d]imidazole.
[0083] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is 1H-benzo[d]imidazole
which is substituted with methyl pyridine or pyrazine.
[0084] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is thiazole which is
substituted with methyl or amino.
[0085] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is imidazole which is
substituted with methyl.
[0086] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is pyrazole which is
substituted with isopropyl.
[0087] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is isoxazole which is
substituted with methyl.
[0088] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is 1,3,4-oxadiazole which
is substituted with methyl.
[0089] According to yet another embodiment there is provided a
compound of formula (1A), wherein R.sub.7 is pyrimidine.
[0090] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is
pyrazolo[1,5-a]pyrimidine.
[0091] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is --S(O).sub.2R.sub.b;
wherein R.sub.b is methyl or 4-chloro phenyl.
[0092] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is N,N-dimethyl
amino.
[0093] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is tertiary butyl.
[0094] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is tertiary butoxy.
[0095] According to yet another embodiment, there is provided a
compound of formula (1A), wherein R.sub.7 is cyclohexyl.
[0096] According to yet another embodiment, there is provided a
compound of formula (1A), wherein `n` is 0.
[0097] According to yet another embodiment, there is provided a
compound of formula (1A), wherein `n` is 1.
[0098] According to yet another embodiment, there is provided a
compound of formula (1A), wherein `n` is 2.
[0099] Accordingly, one other aspect of the present invention
provides compounds of formula (1B):
##STR00006##
wherein,
[0100] R.sub.2, R.sub.5, R.sub.6, R.sub.7 and `n` are same as
defined in claim 1;
[0101] X can be selected from --O--, --CH.sub.2O--, --CH.sub.2N--,
or (--CH.sub.2--).sub.m;
[0102] `m` can be an integer selected from 1, 2, 3 or 4;
[0103] pharmaceutically acceptable salts, pharmaceutically
acceptable solvates, pharmaceutically acceptable hydrates,
tautomers, stereoisomers, ester prodrugs, or combination
thereof.
[0104] It should be understood that the formula (1B) structurally
encompasses all stereoisomers, including enantiomers,
diastereomers, racemates, and combinations thereof which may be
contemplated from the chemical structure of the genus described
herein.
[0105] It should be understood that the formula (1B) structurally
encompasses all tautomers.
[0106] Also contemplated are prodrugs of the compounds of the
formula (1B), including ester prodrugs.
[0107] According to one embodiment, there is provided a compound of
formula (1B), wherein R.sub.2 is hydrogen.
[0108] According to one embodiment, there is provided a compound of
formula (1B), wherein X is --CH.sub.2--.
[0109] According to one embodiment, there is provided a compound of
formula (1B), wherein R.sub.5 and R.sub.6 together represent
oxo.
[0110] According to another embodiment, there is provided a
compound of formula (1B), wherein R.sub.7 is phenyl which is
substituted with chloro, methyl or fluoro.
[0111] According to another embodiment, there is provided a
compound of formula (1B), wherein R.sub.7 is pyridyl which is
substituted with methyl or morpholine.
[0112] According to another embodiment, there is provided a
compound of formula (1B), wherein R.sub.7 is pyrimidine.
[0113] According to another embodiment, there is provided a
compound of formula (1B), wherein `n` is 1.
[0114] According to another embodiment, there is provided a
compound of formula (1B), wherein `m` is 1.
[0115] According to another embodiment, there is provided a
compound of formula (1B), wherein `m` is 2.
[0116] According to another embodiment, there is provided a
compound of formula (1B), wherein `m` is 3.
[0117] According to another embodiment, there is provided a
compound of formula (1B), wherein `m` is 4.
[0118] Below are the representative compounds, which are
illustrative in nature only and are not intended to limit to the
scope of the invention (Nomenclature has been generated from
ChemBioDraw Ultra 13.0 version): [0119]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chlorobe-
nzamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)-
carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound 1),
[0120]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(1-(4-chlorophenyl-
)
cyclopropane-1-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,1-
1a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobut-
ane-1-carboxylic acid (Compound 2), [0121]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-benzamido-2-methyl
propanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,11,
11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl-
)-2,2-dimethyl cyclobutane-1-carboxylic acid (Compound 3), [0122]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(3,4-dichlorobenza-
mido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3-
a,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound 4),
[0123]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(pyrazine-2-carboxamido)propanamido)-2-oxo-3,-
3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 5), [0124]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(6-aminonicotinami-
do)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,-
4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound 6),
[0125]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(5-methylpyrazine-2-carboxamido)propanamido)--
2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (Compound 7), [0126]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-1-(tert-butox-
y
carbonyl)pyrrolidine-2-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,-
5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13-
,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl-
cyclobutane-1-carboxylic acid (Compound 8), [0127]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(4-ethylpiperaz-
in-1-yl)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamet-
hyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro--
2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carb-
oxylic acid (Compound 9), [0128]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-(piperidin-1-yl)acetamido)propanamido)-2-o-
xo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 10), [0129]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-amino-2-methyl
propanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride (Compound 11), [0130]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(1H-benzo[d]imidaz-
ole-5-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamet-
hyl-2-oxo-3,3a,4,
5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]-
chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (Compound 12), [0131]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-(6-methylpyridin-3-yl)-1H-benzo[d]imidazol-
e-5-carboxamido)propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b-
,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-di-
methylcyclobutane-1-carboxylic acid (Compound 13), [0132]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2,4-dimethylthiaz-
ole-5-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamet-
hyl-2-oxo-3,3a,4,
5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]-
chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (Compound 14), [0133]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-(pyrazin-2-yl)-1H-benzo[d]imidazole-5-carb-
oxamido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (Compound 15), [0134]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(1-methyl-1H-imidazole-2-carboxamido)propanam-
ido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-
-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carb-
oxylic acid (Compound 16), [0135]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-3a-(2-(3-iso-
propyl-1H-pyrazole-5-carboxamido)-2-methylpropanamido)-5a,5b,8,8,11a-penta-
methyl-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (Compound 17), [0136]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-morpholinobenzamido)propanamido)-2-oxo-3,3-
a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 18), [0137]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(3,5-dimethylisoxa-
zole-4-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentame-
thyl-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (Compound 19), [0138]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(4-methyl-1H-imidazol-1-yl)benzamido)propa-
namido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-c-
arboxylic acid (Compound 20), [0139]
(1S,3R)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chlorobenzamido-
)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,-
5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)-
carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound 21),
[0140]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-fluorobenzamido-
)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,-
5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound
22), [0141]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b-
,8,8,11a-pentamethyl-3a-(2-methyl-2-(4-methylbenzamido)propanamido)-2-oxo--
3,3a,4,5,5a,5b,6,7,
7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl-
)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound
23), [0142]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(furan-3-ca-
rboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-ox-
o-3,3a,4,5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound
24), [0143]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b-
,8,8,11a-pentamethyl-3a-(2-methyl-2-(4-(trifluoromethyl)benzamido)propanam-
ido)-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 25), [0144]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(furan-2-carboxami-
do)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,-
4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound
26), [0145]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b-
,8,8,11a-pentamethyl-3a-(2-methyl-2-(1-phenylcyclopentane-1-carboxamido)pr-
opanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadec-
ahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane--
1-carboxylic acid (Compound 27), [0146]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(quinoline-2-carboxamido)propanamido)-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 28), [0147]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(3-methylpicolinamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 29), [0148]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-methylfuran-3-carboxamido)propanamido)-2-o-
xo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 30), [0149]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-morpholinonicotinamido)propanamido)-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 31), [0150]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(pyrimidine-2-carboxamido)propanamido)-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 32), [0151]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2,5-dimethylfuran-
-3-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-
-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 33), [0152]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(1,1-dioxidothi-
o
morpholino)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pen-
tamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecah-
ydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1--
carboxylic acid (Compound 34), [0153]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(piperidine-4-carboxamido)propanamido)-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride (Compound 35), [0154]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-2-amino-3-met-
hyl
butanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-
-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride (Compound 36), [0155]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 37), [0156]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclopropane-1-carboxamido)-2-ox-
o-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 38), [0157]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-fluorobenzamido-
)
cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 39), [0158]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(4-methylbenzamido)cyclopropane-1-carboxamido)-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 40), [0159]
(1S,3R)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 41), [0160]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclobutane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 42), [0161]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclobutane-1-carboxamido)-2-oxo-
-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 43), [0162]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-2-oxo-3a-(1-(pyrimidine-2-carboxamido)cyclobutane-1-carboxam-
ido)-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 44), [0163]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(2-morpholinonicotinamido)cyclobutane-1-carboxamido)-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 45), [0164]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclopentane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 46),
[0165]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b-
,8,8,11a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclopentane-1-carboxamid-
o)-2-oxo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 47), [0166]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclohexane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 48), [0167]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclohexane-1-carboxamido)-2-oxo-
-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 49), [0168]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(4-methylbenzamido)cyclohexane-1-carboxamido)-2-oxo-3,-
3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 50), [0169]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(methylsulfonyl)benzamido)propanamido)-2-o-
xo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 51), [0170]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((S)-piperidine-3-carboxamido)propanamido)-2--
oxo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride (Compound 52), [0171]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(4-chlorophenyl-
)
acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2--
oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 53), [0172]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)prop-
anamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecah-
ydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1--
carboxylic acid (Compound 54), [0173]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-aminothiazole-4-
-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 55), [0174]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(5-methyl-1,3,4-oxadiazol-2-yl)benzamido)p-
ropanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octade-
cahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-
-1-carboxylic acid (Compound 56), [0175]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-(1,1-dioxidothi-
o
morpholino)benzamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pen-
tamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecah-
ydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1--
carboxylic acid (Compound 57), [0176]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-((1,1-dioxidoth-
io
morpholino)methyl)benzamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8-
,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a--
octadecahydro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (Compound 58), [0177]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(dimethylamino)
acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-o-
xo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 59), [0178]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(6-methylpicolinamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 60), [0179]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(6-methylnicotinamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 61), [0180]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-pivalamidopropanamido)-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,11,
11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl-
)-2,2-dimethyl cyclobutane-1-carboxylic acid (Compound 62), [0181]
sodium
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,
11a-pentamethyl-3a-(2-methyl-2-(methylsulfonamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylate (Compound
63), [0182]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((ethoxycar-
bonyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2--
oxo-3,3a,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound
64), [0183]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((4-chlorop-
henyl)
sulfonamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentame-
thyl-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 65), [0184]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(cyclohexanecarbox
amido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 66), [0185]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((pyridin-2-ylmethyl)amino)propanamido)-2-oxo-
-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 67), [0186]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((4-chlorobenzyl)a-
mino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3-
a,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound
68), [0187]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(((1-(4-chl-
orophenyl)
cyclopropyl)methyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5-
b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,-
13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylc-
yclobutane-1-carboxylic acid (Compound 69), [0188]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(5-chloropicolinam-
ido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a-
,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)ox-
y)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid (Compound
70), [0189]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b-
,8,8,11a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-3-yl)ureido)propan-
amido)-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 71), [0190]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-2-yl)ureido)propanamido)--
2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 72), [0191]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((2-(dimethylamino-
)
ethyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl--
2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 73), [0192]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylprop-
an
amido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7-
a,8,9,10,11,11a,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,-
2-dimethyl cyclobutane-1-carboxylic acid hydrochloride (Compound
74), [0193]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-buto-
xycarbonyl)
amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(Compound 75), [0194]
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((S)-pyrrolidine-2-carboxamido)propanamido)-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride (Compound 76), or pharmaceutically acceptable salts,
solvates, including hydrates and prodrugs of compounds are also
contemplated.
[0195] The present invention also provides a pharmaceutical
composition that includes at least one compound as described herein
and at least one pharmaceutically acceptable excipient (such as a
pharmaceutically acceptable carrier or diluent). Specifically, the
pharmaceutical composition comprises a therapeutically effective
amount of at least one compound described herein. The compound(s)
present in the composition may be associated with a
pharmaceutically acceptable excipient (such as a carrier or a
diluent) or may be diluted by a carrier, or enclosed within a
carrier which may be in the form of a capsule, sachet, or other
container.
[0196] The compounds and pharmaceutical compositions described
herein are useful in the treatment of diseases, conditions and/or
disorders mediated by viral infections.
[0197] The present invention further provides a method of treating
a disease, condition and/or disorder mediated by viral infections
in a subject in need thereof by administering to the subject one or
more compounds described herein in a therapeutically effective
amount to cause that infection, specifically in the form of a
pharmaceutical composition.
[0198] Also provided herein are processes for preparing compounds
described herein.
[0199] The invention provides a method for preventing; ameliorating
or treating a HIV mediated disease, disorder or syndrome in a
subject in need thereof comprising administering to the subject a
therapeutically effective amount of a compound of the invention.
The invention further provides a method, wherein the HIV mediated
disease, disorder or syndrome is like AIDS, AIDS related complex,
or a syndrome characterized by symptoms such as persistent
generalized lymphadenopathy, fever and weight loss, or a retroviral
infection genetically related to AIDS.
[0200] Anti HIV inhibitory potential of the compounds of present
invention may be demonstrated by any one or more methodologies
known in the art, such as by using the assays described in Mossman
T, December 1983, Journal of immunological methods, 65 (1-2), 55-63
and S P C Cole, cancer chemotherapy and Pharmacology, 1986, 17,
259-263.
DETAILED DESCRIPTION OF THE INVENTION
[0201] The present invention provides C-3 novel triterpenone with
C-28 reverse amide derivatives and related compounds, which may be
used as antiviral particularly as anti-HIV compounds and processes
for the synthesis of these compounds. Pharmaceutically acceptable
salts, pharmaceutically acceptable solvates, enantiomers,
diastereomers of the derivatives, together with pharmaceutically
acceptable carriers, excipients or diluents, which can be used for
the treatment of diseases, condition and/or disorders mediated by
viral infections, are also provided.
[0202] The following definitions apply to the terms as used
herein:
[0203] The terms "halogen" or "halo" includes fluorine, chlorine,
bromine, or iodine.
[0204] The term "alkyl" refers to a straight or branched
hydrocarbon chain radical consisting solely of carbon and hydrogen
atoms, containing no unsaturation, having from one to eight carbon
atoms, and which is attached to the rest of the molecule by a
single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl
(isopropyl), n-butyl, n-pentyl, and 1,1-dimethylethyl
(t-butyl).
[0205] The term "alkoxy" refers to a straight or branched
hydrocarbon chain with oxygen radical consisting carbon and
hydrogen atoms, containing saturation or unsaturation, having from
one to eight carbon atoms, and which is attached through oxygen
atom to the rest of the molecule by a single bond, e.g., methyloxy,
ethyloxy, n-propyloxy, 1-methylethyloxy (isopropyloxy), n-butyloxy,
n-pentyloxy, and 1,1-dimethylethyloxy (t-butyloxy).
[0206] The term "alkoxylalkoxy" refers to a straight or branched
hydrocarbon chain with oxygen radical consisting carbon atom,
hydrogen atom and alkoxy groups, containing saturation or
unsaturation, having from one to eight carbon atoms, and which is
attached through oxygen atom to the rest of the molecule by a
single bond, e.g., 2-(methyloxy)ethyloxy, 2-(ethyloxy)ethyloxy,
2-(n-propyloxy)ethyloxy, and 3-(isopropyloxy)butyloxy.
[0207] The term "amine" refers to an organic compounds and
functional groups that contain a basic nitrogen atom with a lone
pair. Amines are derivatives of ammonia, wherein one or more
hydrogen atoms have been replaced by a substituent such as an alkyl
or aryl group these may respectively be called alkylamines and
arylamines; amines in which both types of substituent are attached
to one nitrogen atom may be called alkylarylamines. Important
amines include amino acids, trimethylamine, and aniline.
[0208] The term "amino acid" refers to a straight or branched
hydrocarbon chain containing an amine group, a carboxylic acid
group, and a side-chain that is specific to each amino acid and
which is attached through the nitrogen atom of the amine group to
the rest of the molecule by a single bond, e.g., alanine, valine,
isoleucine, leucine, phenylalanine, or tyrosine.
[0209] The term "amino alkyl" refers to any amino derivative of an
alkyl radical more specifically dimethylamino.
[0210] The term "cycloalkyl" denotes a non-aromatic mono or
multicyclic ring system of from 3 to about 12 carbon atoms, such as
cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Examples of
multicyclic cycloalkyl groups include, but are not limited to,
perhydronapththyl, adamantyl and norbornyl groups, bridged cyclic
groups and spirobicyclic groups, e.g., spiro (4,4) non-2-yl.
[0211] The term "aryl" refers to an aromatic radical having from 6
to 14 carbon atoms such as phenyl, naphthyl, tetrahydronapthyl,
indanyl, and biphenyl.
[0212] The term "haloalkyl" refers to alkyl group (as defined
above) is substituted with one or more halogens. A monohaloalkyl
radical, for example, may have a chlorine, bromine, iodine or
fluorine atom. Dihalo and polyhaloalkyl radicals may have two or
more of the same or different halogen atoms. Examples of haloalkyl
include, but are not limited to, chloromethyl, dichloromethyl,
trichloromethyl, dichloroethyl, dichloropropyl, fluoromethyl,
difluoromethyl, trifluoromethyl, pentafluoroethyl,
heptafluoropropyl, difluoro chloromethyl, dichloro fluoromethyl,
difluoroethyl, difluoropropyl and the like.
[0213] The terms "heterocyclyl" and "heterocyclic ring" refer to a
stable 3- to 15-membered ring radical which consists of carbon
atoms and from one to five heteroatoms selected from nitrogen,
phosphorus, oxygen and sulfur. For purposes of this invention, the
heterocyclic ring radical may be a monocyclic, bicyclic or
tricyclic ring system, which may include fused, bridged or spiro
ring systems, and the nitrogen, phosphorus, carbon, oxygen or
sulfur atoms in the heterocyclic ring radical may be optionally
oxidized to various oxidation states. In addition, the nitrogen
atom may be optionally quaternized; and the ring radical may be
partially or fully saturated (i.e., heterocyclic or heteroaryl).
Examples of such heterocyclic ring radicals include, but are not
limited to, tetrazoyl, tetrahydroisouinolyl, piperidinyl,
piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl,
2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl,
pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl,
oxazolidinyl, triazolyl, isoxazolyl, isoxasolidinyl, morpholinyl,
thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl,
isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl,
octahydroindolyl, octahydroisoindolyl, quinolyl, isoquinolyl,
decahydroisoquinolyl, benzimidazolyl, thiadiazolyl, benzothiazolyl,
benzooxazolyl, furyl, tetrahydrofurtyl, tetrahydropyranyl, thienyl,
benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide,
thiamorpholinyl sulfone, dioxaphospholanyl, oxadiazolyl and
1,4-Thiazine-1,1-dione. The heterocyclic ring radical may be
attached to the main structure at any heteroatom or carbon atom
that results in the creation of a stable structure.
[0214] The term "heterocyclylalkyl" refers to a heterocyclic ring
radical directly bonded to an alkyl group. The heterocyclylalkyl
radical may be attached to the main structure at any carbon atom in
the alkyl group that results in the creation of a stable
structure.
[0215] The term "heteroaryl" refers to an aromatic heterocyclic
ring radical. Examples of such heteroaryl include, but are not
limited to pyridyl, pyrazinyl, furanyl, quinolinyl, tetrazoyl,
triazolyl, 1,3-Diaza-1H-indenyl, thiazolyl, imidazolyl, pyrazolyl,
isoxazolyl, pyrazolo[1,5-a]pyrimidinyl, 1,3,4-Oxadiazolyl,
thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl,
indolyl, isoindolyl, indolinyl and isoindolinyl. The heteroaryl
ring radical may be attached to the main structure at any
heteroatom or carbon atom that results in the creation of a stable
structure.
[0216] "Substituted" refers to 1-3 substituents on the same
position or on different positions with the same groups or
different groups. Unless otherwise specified, the term
"substituted" as used herein refers to substitution with any one or
any combination of the following substituents: hydroxy, halogen,
carboxyl, cyano, nitro, oxo (.dbd.O), thio (.dbd.S), substituted or
unsubstituted alkyl, haloalkyl, substituted or unsubstituted
alkoxy, substituted or unsubstituted alkenyl, substituted or
unsubstituted alkynyl, substituted or unsubstituted aryl,
substituted or unsubstituted arylalkyl, substituted or
unsubstituted cycloalkyl, s substituted or unsubstituted amino,
substituted or unsubstituted heteroaryl, substituted or
unsubstituted heterocyclylalkyl ring, substituted or substituted or
unsubstituted heterocyclic ring. The substituents in the
aforementioned "substituted" groups cannot be further substituted.
For example, when the substituent on "substituted alkyl" is
"substituted aryl", the substituent on "substituted aryl" cannot be
"substituted alkenyl".
[0217] The term "prodrug" denotes a derivative of a compound, which
derivative, when administered to warm-blooded animals, e.g. humans,
is converted into the compound (drug). The enzymatic and/or
chemical hydrolytic cleavage of the compounds of the present
invention occurs in such a manner that the proven drug form (parent
carboxylic acid drug) is released, and the moiety or moieties split
off remain nontoxic or are metabolized so that nontoxic metabolic
products are produced. For example, a carboxylic acid group can be
esterified, e.g., with a methyl group or ethyl group to yield an
ester. When an ester is administered to a subject, the ester is
cleaved, enzymatically or non-enzymatically, reductively,
oxidatively, or hydrolytically, to reveal the anionic group. An
anionic group can be esterified with moieties (e.g., acyloxymethyl
esters) which are cleaved to reveal an intermediate compound which
subsequently decomposes to yield the active compound. A discussion
of the use of prodrugs is provided by T. Higuchi and W. Stella,
"Pro-drugs as Novel Delivery Systems," Vol. 14 of the A.C.S.
Symposium Series, and in Bioreversible Carriers in Drug Design, ed.
Edward B. Roche, American Pharmaceutical Association and Pergamon
Press, 1987.
[0218] The term "treating" or "treatment" of a state, disease,
disorder or condition includes: [0219] (1) preventing or delaying
the appearance of clinical symptoms of the state, disease, disorder
or condition developing in a subject that may be afflicted with or
predisposed to the state, disease, disorder or condition but does
not yet experience or display clinical or subclinical symptoms of
the state, disease, disorder or condition; [0220] (2) inhibiting
the state, disease, disorder or condition, i.e., arresting or
reducing the development of the state, disease, disorder or
condition or at least one clinical or subclinical symptom thereof;
or [0221] (3) relieving the state, disease, disorder or condition,
i.e., causing regression of the state, disease, disorder or
condition or at least one of its clinical or subclinical
symptoms.
[0222] The benefit to a subject receiving treatment is either
statistically significant or at least perceptible to the subject or
to the physician.
[0223] The term "subject" includes mammals (especially humans) and
other animals, such as domestic animals (e.g., household pets
including cats and dogs) and non-domestic animals (such as
wildlife).
[0224] A "therapeutically effective amount" means the amount of a
compound that, when administered to a subject for treating a state,
disease, disorder or condition, is sufficient to effect such
treatment. The "therapeutically effective amount" will vary
depending on the compound, the state, disease, disorder or
condition and its severity and the age, weight, physical condition
and responsiveness of the subject receiving treatment.
[0225] The compounds of the present invention may form salts.
Non-limiting examples of pharmaceutically acceptable salts forming
part of this invention include salts derived from inorganic bases
salts of organic bases salts of chiral bases, salts of natural
amino acids and salts of non-natural amino acids. Certain compounds
of the present invention are capable of existing in stereo isomeric
forms (e.g., diastereomers, enantiomers, racemates, and
combinations thereof). With respect to the overall compounds
described by the Formula (1), the present invention extends to
these stereo isomeric forms and to mixtures thereof. To the extent
prior art teaches synthesis or separation of particular
stereoisomers, the different stereo isomeric forms of the present
invention may be separated from one another by the methods known in
the art, or a given isomer may be obtained by stereospecific or
asymmetric synthesis. Tautomeric forms and mixtures of compounds
described herein are also contemplated.
[0226] Pharmaceutically acceptable solvates includes hydrates and
other solvents of crystallization (such as alcohols). The compounds
of the present invention may form solvates with low molecular
weight solvents by methods known in the art.
Pharmaceutical Compositions
[0227] The pharmaceutical compositions provided in the present
invention include at least one compound described herein and at
least one pharmaceutically acceptable excipient (such as a
pharmaceutically acceptable carrier or diluent). Specifically, the
contemplated pharmaceutical compositions include a compound(s)
described herein in an amount sufficient to treat viral infection
in a subject.
[0228] The subjects contemplated include, for example, a living
cell and a mammal, including human. The compound of the present
invention may be associated with a pharmaceutically acceptable
excipient (such as a carrier or a diluent) or be diluted by a
carrier, or enclosed within a carrier which can be in the form of a
capsule, sachet, or other container.
[0229] Examples of suitable carriers include, but are not limited
to, water, salt solutions, alcohols, polyethylene glycols, peanut
oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin,
magnesium carbonate, sugar, amylose, magnesium stearate, talc,
gelatin, agar, pectin, acacia, stearic acid, lower alkyl ethers of
cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid
monoglycerides and diglycerides, fatty acid esters, and
polyoxyethylene.
[0230] The carrier or diluent may include a sustained release
material, such as, for example, glyceryl monostearate or glyceryl
distearate, alone or mixed with a wax.
[0231] The pharmaceutical composition may also include one or more
pharmaceutically acceptable auxiliary agents, wetting agents,
emulsifying agents, suspending agents, preserving agents, salts for
influencing osmotic pressure, buffers, sweetening agents, flavoring
agents, colorants, or any combination of the foregoing. The
pharmaceutical composition of the invention may be formulated so as
to provide quick-, sustained-, or delayed-release of the active
ingredient after administration to the subject by employing
procedures known in the art.
[0232] The pharmaceutical compositions described herein may be
prepared, e.g., as described in Remington: The Science and Practice
of Pharmacy, 20.sup.th Ed., 2003 (Lippincott Williams &
Wilkins). For example, the active compound can be mixed with a
carrier, or diluted by a carrier, or enclosed within a carrier,
which may be in the form of an ampule, capsule, or sachet. When the
carrier serves as a diluent, it may be a solid, semi-solid, or
liquid material that acts as a vehicle, excipient, or medium for
the active compound.
[0233] The pharmaceutical compositions may be in conventional
forms, for example, capsules, tablets, solutions, suspensions,
injectables or products for topical application. Further, the
pharmaceutical composition of the present invention may be
formulated so as to provide desired release profile.
[0234] The route of administration may be any route which
effectively transports the active compound to the appropriate or
desired site of action. Suitable routes of administration include,
but are not limited to, oral, nasal, pulmonary, buccal, subdermal,
intradermal, transdermal, parenteral, rectal, depot, subcutaneous,
intravenous, intraurethral, intramuscular, intranasal, ophthalmic
(such as with an ophthalmic solution) or topical (such as with a
topical ointment). The oral route is specifically suitable.
[0235] Solid oral formulations include, but are not limited to,
tablets, capsules (soft or hard gelatin), dragees (containing the
active ingredient in powder or pellet form), troches and lozenges.
Tablets, dragees, or capsules having talc and/or a carbohydrate
carrier or binder or the like are particularly suitable for oral
application. Exemplary carriers for tablets, dragees, or capsules
include lactose, cornstarch, and/or potato starch. A syrup or
elixir can be used in cases where a sweetened vehicle can be
employed.
[0236] A typical tablet that may be prepared by conventional
tableting techniques.
[0237] Liquid formulations include, but are not limited to, syrups,
emulsions, soft gelatin and sterile injectable liquids, such as
aqueous or non-aqueous liquid suspensions or solutions.
[0238] For parenteral application, particularly suitable are
injectable solutions or suspensions, specifically aqueous solutions
with the active compound dissolved in polyhydroxylated castor
oil.
Methods of Screening
[0239] Antiviral HIV activity and cytotoxicity of compounds present
invention can be measured in parallel by following the methods
published in the literature.
[0240] The cytotoxic effect of compounds can be analyzed by
measuring the proliferation of cells using the
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazlium bromide (MTT)
staining. Cells (5.times.10.sup.3 cells/well) will be incubated in
96 well plates in the presence or absence of compounds. At the end
of treatment, 20 .mu.l of MTT (5 mg/ml in PBS) will be added to
each well and incubated for an additional 4 hours at 37.degree. C.
The purple-blue MTT formazan precipitate will be dissolved in a
triplex reagent containing 10% SDS, 5% isobutanol and 10 mmol/lit
HCl. The activity of mitochondria, reflecting cellular growth and
viability, will be evaluated by measuring the optical density at
570 nm on micro titer plate.
[0241] Action of compounds on replication of HIV in Sup-T1 cells
can be determined by the method published by Roda Rani et al., 2006
(Archives of Biochemistry and Biophysics, Volume 456, Issue 1, 1
Dec. 2006, Pages 79-92).
[0242] Briefly, 1.times.10.sup.6 Sup-T1 cells with 100% cell
viability will be seeded in RPMI 1640, 0.1% FBS four 12 well
plates. Increasing concentrations of Epap-1 peptides will be added
to the cells and will be infected with HIV1.sub.93 IN 101 each at
final concentration of virus equivalent to 2 ng of p24 per ml. The
infected cells will be incubated at 37.degree. C. and 5% CO.sub.2
incubator for 2 hours. After 2 hours the cells will be pelleted at
350 g for 10 minutes, supernatant will be discarded and cell will
be held with RPMI 1640 containing 10% FBS. The cells will be
resuspended in the same medium with increasing concentrations of
Epap-1 peptides and will be incubated for 96 hours. The cells will
be supplemented with peptides at every 24 hours. The supernatants
will be collected after 96 hours and analyzed using P24 antigen
capture assay kit (SAIC Fredrick). The infection in the absence of
Epap-1 will be considered to be 0% inhibition Azidothymidine (AZT)
will be taken as positive control.
[0243] Action of compound on virus entry and quantification of
virus entered can be done in terms of GFP expression by the
following the methods published J. Virol. 72, 6988 (1998) by in
Cecilia et al., and Analytical Biochemistry Volume 360, Issue 2, 15
Jan. 2007, Pages 315-317 (Dyavar S. Ravi and Debashis Mitra).
[0244] Briefly, cells will be seeded in to wells of 24 well plates
1 day prior to the experiment. The cells will be transfected with
Tat-reporter. The virus inoculum will be adjusted to 1,000-4,000
TCID 50/ml in assay medium (DMEM, 10% FCS, glutamine and
antibiotics), 50 .mu.l aliquots will be incubated with serial
dilutions of compounds (50 .mu.l) for 1 hour at 37.degree. C. The
reporter expression will be quantified at appropriate time
calculated inhibitory doses referrers to the concentration of these
agents in this preincubation mixture.
[0245] Other relevant references useful for screening antiviral HIV
activity are: Averett, D. R. 1989. Anti-HIV compound assessment by
two novel high capacity assays. J. Virol. Methods 23: 263-276;
Schwartz, O., et al. 1998; A rapid and simple colorimeric test for
the study of anti HIV agents. AIDS Res. and Human Retroviruses,
4(6):441-447; Daluge, S. M., et al. 1994.
5-Chloro-2',3'-deoxy-3'fluorouridine (935U83), a selective anti
human immunodeficiency virus agent with an improved metabolic and
toxicological profile; Antimicro. Agents and Chemotherapy,
38(7):1590-1603; H. Mitsuya and S. Border, Inhibition of the in
vitro infectivity and cytopathic effect of human T-lymphotropic
virus type lymphadenopathy-associated virus (HLTV-III/LAV) by
2,3'-dideoxynucleosides, Proc. Natl. Acad. Sci. USA, 83,
1911-15(1986); Pennington et al., Peptides 1990; Meek T. D et al.,
Inhibition of HIV-1 protease in infected T-limphocytes by synthetic
peptide analogues, Nature, 343, p 90 (1990); Weislow et al., J.
Natl. Cancer Inst. 81, 577-586, 1989; T. Mimoto et al., J. Med.
Chem., 42, 1789-1802, 1999; Uckun et al 1998, Antimicrobial Agents
and Chemotherapy 42:383; for P24 antigen assay Erice et al., 1993,
Antimicrob. Ag. Chemotherapy 37: 385-383; Koyanagi et al., Int. J.
Cancer, 36, 445-451, 1985; Balzarini et al. AIDS (1991), 5, 21-28;
Connor et al., Journal of virology, 1996, 70, 5306-5311; Popik et
al., Journal of virology, 2002, 76, 4709-4722; Harrigton et al.,
Journal of Virology Methods, 2000, 88, 111-115; Roos et al.,
Virology 2000, 273, 307-315; Fedyuk N. V. et al; Problems of
Virology 1992, (3)P135; Mosmann T, December 1983, Journal of
immunological methods, 65 (1-2), 55-63; SPC Cole, cancer
chemotherapy and Pharmacology, 1986, 17, 259-263, Antiviral methods
and protocols (Eds: D Kinchington and R. F. Schinazi) Humana Press
Inc., 2000, HIV protocols (Eds: N. L. Michael and J. H. Kim) Humana
Press Inc, 1999, DAIDS Virology manual from HIV laboratories,
Publication NIH-97-3838, 1997, 4. HIV-1 p24 antigen capture assay,
enzyme immunoassay for detection of Human immunodeficiency Virus
Type 1 (HIV-1) p24 in tissue culture media--Advanced bio science
laboratories, Inc kit procedure.
Methods of Treatment
[0246] The present invention provides compounds and pharmaceutical
formulations thereof that are useful in the treatment of diseases,
conditions and/or disorders mediated by viral infections. The
connection between therapeutic effect and antiviral is illustrated.
For example, PCT publication Nos. WO 01/07646, WO 01/65957, or WO
03/037908; US publication Nos. U.S. Pat. No. 4,598,095 or US
2002/0068757; EP publication Nos. EP 0989862 or EP 0724650;
Bioorganic & Medicinal Chemistry Letters, 16, (6), 1712-1715,
2006; and references cited therein, all of which are incorporated
herein by reference in their entirety and for the purpose
stated.
[0247] The present invention further provides a method of treating
a disease, condition and/or disorder mediated by viral infections
in a subject in need thereof by administering to the subject a
therapeutically effective amount of a compound or a pharmaceutical
composition of the present invention.
[0248] Diseases, conditions, and/or disorders that are mediated by
viral infections are believed to include, but are not limited to,
HIV infection, HBV infection, HCV infection, a retroviral infection
genetically related to HIV, AIDS, inflammatory disease, respiratory
disorders (including adult respiratory distress syndrome (ARDS),
bronchitis, chronic bronchitis, chronic obstructive pulmonary
disease, cystic fibrosis, asthma, emphysema, rhinitis and chronic
sinusitis), inflammatory bowel disease (including Crohn's disease
and ulcerative colitis), multiple sclerosis, rheumatoid arthritis,
graft rejection (in particular but not limited to kidney and lung
allografts), endometriosis, type I diabetes, renal diseases,
chronic pancreatitis, inflammatory lung conditions, chronic heart
failure and bacterial infections (in particular but not limited to
tuberculosis).
[0249] The compounds of the present invention can obtain more
advantageous effects than additive effects in the prevention or
treatment of the above diseases when using suitably in combination
with the available drugs. Also, the administration dose can be
decreased in comparison with administration of either drug alone,
or adverse effects of co administrated drugs other than antiviral
can be avoided or declined.
Methods of Preparation
[0250] The compounds described herein may be prepared by techniques
known in the art. In addition, the compounds described herein may
be prepared by following the reaction sequence as depicted in
Scheme 1. Further, in the following schemes, where specific bases,
acids, reagents, solvents, coupling agents, etc., are mentioned, it
is understood that other bases, acids, reagents, solvents, coupling
agents etc., known in the art may also be used and are therefore
included within the present invention. Variations in reaction
conditions, for example, temperature and/or duration of the
reaction, which may be used as known in the art, are also within
the scope of the present invention. All the stereoisomers of the
compounds in these schemes, unless otherwise specified, are also
encompassed within the scope of this invention.
[0251] Compounds of the present invention can be synthesized from
naturally occurring Betulinic acid or betulinal. Key intermediates
required for synthesizing analogues are either commercially
available, or can be prepared by the methods published in the
literature. For example, the key intermediates in the present
invention were prepared by modifying the procedures published in
Journal of organic chemistry 2010, 75, 1285-1288; Journal of
organic chemistry 2000, 65, 3934-3940; Tetrahedron: asymmetry 2008,
19, 302-308; or Tetrahedron. asymmetry 2003, 14, 217-223.
##STR00007## ##STR00008##
[0252] The compounds of formula 1 (wherein, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6 and R.sub.7 are same as defined above)
can be prepared as described in Scheme 1. The C-3 & C-28 di
alcohol compounds of formula (i) can be reacted with a suitable
acetate forming reagents such as anhydrides, acid halides, mixed
anhydrides or the like in the presence of bases such as
triethylamine (TEA), diisopropylethylamine (DIPEA), pyridine or the
like in the solvents such as dichloromethane (DCM), chloroform
(CHCl.sub.3), toluene, tetrahydrofuran (THF) or the like with or
without addition of catalysts such as dimethyl amino pyridine
(DMAP) or the like to give the C-3 & C-28 protected alcohol
compounds of formula (ii) (P.sub.1 and P.sub.2 are protecting
groups such as acetyl, benzyl or the like). The C-3 & C-28
protected alcohol compounds of formula (ii) with terminal double
bond can be migrated to the E-ring compounds of formula (iii) in
the presence of hydrogen bromide (HBr) in acetic acid (AcOH),
acetic acid (AcOH) and acetic anhydride (Ac.sub.2O) in solvents
like toluene, benzene, xylene or the like. The E-ring compounds of
formula (iii) can be converted to give the Enone compounds of
formula (iv) in the presence of sodium dichromate
(Na.sub.2Cr.sub.2O.sub.7), sodium acetate (NaOAc), acetic acid
(AcOH), acetic anhydride (Ac.sub.2O) in solvents like toluene,
benzene or the like. The Enone C-28 compounds of formula (iv) can
be deprotected to give the C-28 alcohol compounds of formula (v) in
the presence of potassium hydroxide (KOH) or the like in the
combination of solvents such as toluene:ethanol (EtOH) (1:1) or
with reagents like Aluminum isopropoxide (Al(i-Pro).sub.3) in
solvents like 2-propanol or the like. The C-28 alcohol compounds of
formula (v) can be converted to give the C-28 aldehyde compounds of
formula (vi) in the presence of pyridinium chlorochromate (PCC),
pyridinium dichromate (PDC), Dess-martin periodinane (DMP) or Swern
oxidation conditions in the solvents such as dichloromethane (DCM)
or the like. The C-28 aldehyde compounds of formula (vi) can be
converted to give the C-28 acid compounds of formula (vii) in the
presence of oxidising agents such as sodium chlorite (NaClO.sub.2)
or the like in the presence of a scavenger such as
2-methyl-2-butene or the like in the presence of a buffer reagent
such as sodiumdihydrogen phosphate (NaH.sub.2PO.sub.4) or the like
in the combination of solvents such as tert-butanol (t-BuOH),
tetrahydrofuran (THF) and water (H.sub.2O) or the like. The C-28
alcohol compounds of formula (v) can be converted in one pot method
to C-28 acid compounds of formula (vii) in the presence of
oxidizing agents such as 2,2,6,6-Tetramethyl-1-piperidinyloxy, free
radical, 2,2,6,6-Tetramethylpiperidine 1-oxyl (TEMPO), Sodium
hypochlorite (NaOCl) and Sodium chlorite (NaClO.sub.2) in the
presence of buffer reagent such as sodiumdihydrogen phosphate
(NaH.sub.2PO.sub.4) and the bases like NaHCO.sub.3 in combination
of solvents like Tetrahydrofuran (THF) and water (H.sub.2O) The
C-28 acid compounds of formula (vii) can be converted to the C-28
carbamate compounds of formula (viii) by using the reagents like
diphenylphosphoryl azide (DPPA) or ethylchloroformate and sodium
azide (NaN.sub.3) in the presence of bases such as triethylamine
(TEA), N,N-Diisopropylethylamine (DIPEA) in solvents such as
1,2-DCE, THF or Toluene in the presence of alcohols such as
4-methoxybenzyl alcohol (PMBOH), tert-butyl alcohol (t-BuOH) or the
like. The C-28 carbamate compounds of formula (viii) can be cleaved
in the presence of acid medium such as trifluoroacetic acid (TFA),
HCl/1,4-dioxane or the like in the solvents such as dichloromethane
(DCM) or chloroform (CHCl.sub.3) or the like to give the amine
compounds of formula (ix). The C-28 amine compounds of formula (ix)
can be reacted with the acid compounds of formula (x) in the
presence of coupling reagents such as
0-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluroniumhexafluorophosphat-
e (HATU),
O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluroniumhexafluoro
phosphate (HBTU), 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
(EDCI), 1-Hydroxybenzo triazole (HOBt) or the like in the presence
of bases such as triethylamine (TEA), N,N-Diisopropylethylamine
(DIPEA) or the like in the solvents such as 1,2-dichloroethane
(1,2-DCE), dimethylformamide (DMF) or the like to give the C-28
amide compounds of formula (xi). The C-3 protected alcohol
compounds of formula (xi) can be deprotected to give the C-3
alcohol compounds of formula (xii) in the presence of inorganic
bases such as Lithium hydroxide (LiOH), sodium hydroxide (NaOH),
potassium hydroxide (KOH) or the like in the solvents such as
methanol (MeOH):tetrahydrofuran (THF):water (H.sub.2O) (4:2:1) (or)
1,4-dioxane:water (H.sub.2O) (4:1) or the like. The C-3 alcohol
compounds of formula (xii) can be reacted with the acid compounds
of formula (xiii) to give the C-3 ester compounds of formula (xiv)
in different ways like [0253] (a) Acid and alcohol coupling in the
presence of coupling reagents such as 2,4,6-trichlorobenzyl
chloride, or the like in the presence of bases such as
triethylamine (TEA), N,N-Diisopropylethylamine (DIPEA) and
catalysts such as 4-dimethylaminopyridine (DMAP) in the solvents
such as 1,2-dichloroethane (1,2-DCE), dichloromethane (DCM) or the
like. [0254] (b) acid alcohol coupling in the presence of coupling
reagents such EDCI, HOBt in the presence of bases such as
triethylamine (TEA), N,N-Diisopropylethylamine (DIPEA) and
catalysts such as 4-dimethylaminopyridine (DMAP) in the solvents
such as dichloromethane (DCM) and N,N-dimethylformamide (DMF) or
the like.
[0255] The C-28 substituted N-protected compounds of formula (xiv)
can be deprotected in the presence of deprotecting agents such as
trifluoroacetic acid (TFA) or HCl/1,4-dioxane or the like in the
solvents such as dichloromethane (DCM) or the like to give the C-28
substituted amine compounds of formula (xv). The C-28 substituted
amine compounds of formula (xv) can be reacted with the compounds
of formula (xvi) to form C-3 ester compounds of formula (xvii) in
different ways like [0256] a) acid and amine coupling in the
presence of coupling reagents such as
0-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetra
methyluroniumhexafluorophosphate (HATU),
O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyl
uroniumhexafluorophosphate (HBTU),
1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI),
1-Hydroxybenzotriazole (HOBt) or the like in the presence of bases
such as triethylamine (TEA), N,N-Diisopropylethylamine (DIPEA) or
the like in the solvents such as 1,2-dichloroethane (1,2-DCE),
dimethylformamide (DMF) or the like. [0257] b) reductive amination
of amine and aldehyde in the presence of reducing agents such as
sodium triacetoxyborohydride (STAB) or Sodium borohydride or sodium
cyanoborohydride (NaCNBH.sub.3) or the like in the solvents such as
1,2-dichloroethane (1,2-DCE), tetrahydrofuran (THF), Methanol
(MeOH), Acetonitrile (CH.sub.3CN) or the like. [0258] c) acid
chloride and amine coupling in the presence of bases such as
triethylamine (TEA), or diisopropylethylamine (DIPEA) or the like
in the solvents such as dichloromethane (DCM) or the like. [0259]
d) amine and sodium sulphite adduct coupling in the presence of
reductive agents such as sodium cyano borohydride (NaCNBH.sub.3) or
the like in the presence of bases such as triethylamine (TEA) or
the like in the solvents such as methanol (MeOH) or the like.
[0260] e) Amine and isocyanato coupling in the presence of bases
such as triethylamine (TEA), diisopropylethylamine (DIPEA) or the
like in the solvents such as tetrahydrofuran (THF) or the like.
[0261] The ester compounds of formula (xvii) can be hydrolysed to
give the acid compounds of formula 1 in the presence of aqueous
solution of inorganic bases such as Lithium hydroxide (LiOH),
sodium hydroxide (NaOH) or potassium hydroxide (KOH) or the like in
the combination of solvents such as tetrahydrofuran (THF):methanol
(MeOH) (1:1) or the like.
[0262] The abbreviations used in the entire specification may be
summarized herein below with their particular meaning: DIPEA
(N,N-Diisopropylethylamine); .degree. C. (degree Celsius); 6
(delta); ppm (parts per million); % (percentage); DMSO-d.sub.6
(Deuterated DMSO); d (Doublet); dd (Doublet of doublet); EtOH
(Ethanol); EtOAc (Ethyl acetate); g or gr (gram); H or H.sub.2
(Hydrogen); HCl (Hydrochloric acid); h or hr. (Hours); HATU
(O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluroniumhexafluoro
phosphate); Hz (Hertz); HPLC (High-performance liquid
chromatography); mmol (Milli mol); M (Molar); ml (Millilitre); mg
(Milli gram); m (Multiplet); mm (Millimetre); MHz (Megahertz);
ESI-MS (Electron spray Ionization Mass spectra); min (Minutes); mM
(Milli molar); NaOH (Sodium hydroxide); N.sub.2 (Nitrogen); NMR
(Nuclear magnetic resonance spectroscopy); S (Singlet); TEA
(Triethyl amine); TLC (Thin Layer Chromatography); THF
(Tetrahydrofuran); tert (Tertiary), TFA/CF.sub.3COOH (Trifluoro
acetic acid); t (Triplet); IC (Inhibitory concentration), nM (Nano
molar); pH (Pouvoir hydrogen); (Boc).sub.2O (Di-tert-butyl
dicarbonate); DCM (dichloromethane); DMF (N,N-dimethyl formamide);
DMAP (4-(Dimethylamino)pyridine); eq (equivalent); Ltr or L
(Liter); CDCl.sub.3 (Deuterated chloroform); J (Coupling constant);
J.sub.AB (Coupling constant); NaH.sub.2PO.sub.4 (Sodium dihydrogen
phosphate); Na(OAc).sub.3BH.sub.3 (Sodium triacetoxyborohydride);
AcOH (Acetic acid); NaCNBH.sub.3 (Sodium cyanoborohydride); ABq (AB
quartet); Cs.sub.2CO.sub.3 (Cesium carbonate); CuI (Copper(I)
iodide); MTBE (Methyl tert-butyl ether); HBr (Hydrogen bromide);
Ac.sub.2O (Acetic anhydride); NaHCO.sub.3 (Sodium bicarbonate);
Na.sub.2SO.sub.4 (Sodium sulphate); 1,2-DCE (1,2-dichloroethane);
HBTU (O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluroniumhexafluoro
phosphate); KOH (Potassium hydroxide); MeOH (methanol); EDCI
(1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide); HOBt
(1-Hydroxybenzotriazole); brs (broad singlet); DPPA (Diphenyl
phosphoryl azide) and BINAP
(2,2'-bis(diphenylphosphino)-1,1'-binaphthyl).
EXPERIMENTAL
[0263] The present invention is further illustrated by the
following examples, which are not to be construed in any way as
imposing limitations upon the scope of this disclosure, but rather
are intended to be illustrative only. On the contrary, it is to be
clearly understood that resort may be had to various other
embodiments, modifications, and equivalents thereof which, after
reading the description herein, may suggest themselves to one of
ordinary skill in the art without departing from the spirit of the
present invention. Thus, the skilled artisan will appreciate how
the experiments and examples may be further implemented as
disclosed by variously altering the following examples,
substituents, reagents, or conditions.
INTERMEDIATES
Intermediate 1: Preparation of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00009##
[0264] Step 1: Synthesis of
((1R,3aS,5aR,5bR,7aR,9S,11aR,11bR,13aR,13bR)-9-acetoxy-5a,5b,8,8,
11a-pentamethyl-1-(prop-1-en-2-yl)icosahydro-3aH-cyclopenta[a]chrysen-3a--
yl)methyl acetate
##STR00010##
[0266] A mixture of
(1R,3aS,5aR,5bR,7aR,9S,11aR,11bR,13aR,13bR)-3a-(hydroxymethyl)-5a,5b,8,8,-
11a-pentamethyl-1-(prop-1-en-2-yl)icosahydro-1H-cyclopenta[a]chrysen-9-ol
(400 g, 0.904 mol, 1.0 eq) and acetic anhydride (3.4 Ltr) were
heated at 140.degree. C. for about 3 hours. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was cooled to 0.degree. C., solid was filtered,
washed with water (2 Ltr) and dried under vacuum to obtain the
desired product (400 g, yield: 84%) as an off-white solid. .sup.1H
NMR (300 MHz, CDCl.sub.3): .delta. ppm 4.68 (d, 1H), 4.59 (s, 1H),
4.50-4.43 (m, 1H), 4.25 (d, J=11.1 Hz, 1H), 3.85 (d, J=11.1 Hz,
1H), 2.50-2.40 (m, 1H), 2.07 (s, 3H), 2.04 (s, 3H), 2.01-1.71 (m,
4H), 1.70-1.62 (m, 4H), 1.68 (s, 3H), 1.61-1.43 (m, 4H), 1.43-1.36
(m, 4H), 1.33-1.18 (m, 3H), 1.18-1.09 (m, 1H), 1.08-0.94 (m, 3H),
1.03 (s, 3H), 0.97 (s, 3H), 0.88-0.75 (m, 10H).
Step 2. Synthesis of
((3aS,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-acetoxy-1-isopropyl-5a,5b,8,
8,11a-pentamethyl-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octade-
cahydro-3aH-cyclopenta[a]chrysen-3a-yl)methyl acetate
##STR00011##
[0268] HBr in acetic acid (800 ml, 33%), was added to a suspension
of ((1R,3aS,5aR,5bR,
7aR,9S,11aR,11bR,13aR,13bR)-9-acetoxy-5a,5b,8,8,11a-pentamethyl-1-(prop-1-
-en-2-yl) icosahydro-3aH-cyclo penta[a]chrysen-3a-yl)methyl acetate
(step 1, 400 g, 0.76 mol, 1.0 eq) in toluene (800 ml), Ac.sub.2O
(800 ml) and acetic acid (800 ml) previously heated at 105.degree.
C. The reaction mixture was stirred and heated at this temperature
for about 1.5 hours. After cooling down, sodium acetate (480 g) was
added and the resulting reaction mixture was evaporated to dryness.
The residue was taken up in CH.sub.2Cl.sub.2 (1200 ml) and the
organic phase was washed with water (2.times.500 ml), dried over
sodium sulphate, filtered and evaporated under reduced pressure.
The residue was recrystallized over 95% ethanol and
CH.sub.2Cl.sub.2 to obtain the desired product (256 g, yield: 64%)
as a white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm
4.52-4.45 (m, 1H), 4.03 (d, J=10.8 Hz, 1H), 3.98 (d, J=10.8 Hz,
1H), 3.19-3.08 (m, 1H), 2.46-2.38 (m, 1H), 2.28-2.22 (m, 2H), 2.05
(s, 3H), 2.04 (s, 3H), 2.01-1.83 (m, 2H), 1.78-1.63 (m, 6H),
1.57-1.44 (m, 3H), 1.43-1.08 (m, 8H), 1.06 (s, 3H), 1.02-0.88 (m,
12H), 0.84 (s, 3H), 0.83 (s, 3H) and 0.78 (m, 1H).
Step 3: Synthesis of
((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-acetoxy-1-isopropyl-5a,5b,8,
8,11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a--
octadecahydro-3aH-cyclopenta[a]chrysen-3a-yl)methyl acetate
##STR00012##
[0270] To a stirred solution of
((3aS,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-acetoxy-1-isopropyl-5a,5b,8,8,11a--
pentamethyl-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa
decahydro-3aH-cyclopenta[a]chrysen-3a-yl)methyl acetate (step 2,
100 g, 0.190 mol, 1.0 eq) in Toluene (1280 ml) was added sodium
acetate (88.96 g, 1.08 mol, 5.7 eq), sodium dichromate dihydrate
(67.9 g, 0.228 mol, 1.2 eq), Ac.sub.2O (414 ml) and AcOH (1700 ml)
and heated at 60.degree. C. for about 14 hours. TLC indicated
starting material was consumed and the desired product was
observed. After cooling down, the reaction mixture diluted with
water (500 ml) and extracted with ethyl acetate (1000 ml). The
organic phase was washed successively with saturated solution of
sodium carbonate (1.times.500 ml) and brine (2.times.200 ml)
solution. The organic layer was dried over sodium sulphate,
filtered and concentrated under reduced pressure. The residue was
triturated with methanol and the precipitates that formed were
collected by filtration were dried under vacuum to obtain the
desired product (81 g, yield: 79%) as a white solid. .sup.1H NMR
(300 MHz, CDCl.sub.3): .delta. ppm 4.47 (dd, J=10.2, 6.0 Hz, 1H),
4.31 (d, J=10.8 Hz, 1H), 4.03 (d, J=10.8 Hz, 1H), 3.22-3.12 (m,
1H), 2.85 (dd, J=12.3, 3.3 Hz, 1H), 2.36 (d, J=18.6 Hz, 1H), 2.03
(s, 3H), 1.97 (s, 3H), 1.93-1.88 (m, 2H), 1.88-1.62 (m, 6H),
1.61-1.28 (m, 8H), 1.27-1.22 (m, 1H), 1.21-1.12 (m, 9H), 1.09-0.97
(m, 1H), 0.91 (s, 3H), 0.90 (s, 3H), 0.83 (s, 3H), 0.82 (s, 3H) and
0.77-0.75 (m, 1H); ESI-MS: m/z 563.4 (M+Na).sup.+
Step 4: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(hydroxymethyl)-1-isopropyl-5a,5b,-
8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13-
a-octa decahydro-2H-cyclopenta[a]chrysen-9-yl acetate
##STR00013##
[0272] To a stirred solution of
((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-acetoxy-1-isopropyl-5a,5b,8,8,11a--
pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadec-
ahydro-3aH-cyclopenta[a]chrysen-3a-yl)methyl acetate (step 3, 70 g,
0.129 mol, 1.0 eq) in ethanol (2 L):toluene (2 L) was added
potassium hydroxide (8.72 g, 0.155 mol, 1.2 eq) and stirred at room
temperature for about 2 hours. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was neutralized with aqueous 1N HCl to pH adjusted to 7.0 and
evaporated to dryness. The obtained residue was taken up in water
(200 ml) and a small amount of acetone (20 ml). The precipitates
formed were collected by filtration, washed with water and dried in
vacuo to obtain the desired product (51 g, yield: 79%) as a white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 4.49 (dd,
J=10.5, 6.0 Hz, 1H), 3.73 (d, J=10.5 Hz, 1H), 3.67 (d, J=10.5 Hz,
1H), 3.25-3.14 (m, 1H), 2.78 (dd, J=12.3, 3.0 Hz, 1H), 2.43 (d,
J=18.6 Hz, 1H), 2.05 (s, 3H), 2.02-1.65 (m, 8H), 1.60-1.25 (m, 8H),
1.24-1.17 (m, 7H), 1.13 (s, 3H), 1.12-0.97 (m, 1H), 0.94 (s, 3H),
0.92 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H), 0.80 (m, 1H); ESI-MS: m/z
521.3 (M+Na).sup.+.
Step 5: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-formyl-1-isopropyl-5a,5b,8,
8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-
-octadecahydro-2H-cyclopenta[a]chrysen-9-yl acetate
##STR00014##
[0274] To a solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(hydroxymethyl)-1-isopropyl-5a,5b,-
8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,
13a-octadecahydro-2H-cyclopenta[a]chrysen-9-ylacetate (step 4, 52.0
g, 0.104 mol, 1.0 eq) in CH.sub.2Cl.sub.2 (2 L) at room temperature
was added pyridiniumchlorochromate (67.5 g, 0.313 mol, 3.0 eq) and
silicagel (100-200 mesh) (67.5 g). The reaction mixture was stirred
at room temperature for about 1 hour. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was diluted with water (50 ml) and extracted with
CH.sub.2Cl.sub.2. The combined organic layers were washed with
saturated sodium bicarbonate solution, dried over sodium sulphate
and evaporated under reduced pressure to give a crude product,
which was triturated with ethanol, solid was filtered and dried
under vacuum to obtain the desired product (41.4 g, yield: 80%) as
a white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 9.31
(s, 1H), 4.52-4.44 (m, 1H), 3.32-3.18 (m, 1H), 2.60-2.50 (m, 1H),
2.43-2.33 (m, 2H), 2.12-2.0 (m, 2H), 2.05 (s, 3H), 2.0-1.80 (m,
2H), 1.80-1.65 (m, 3H), 1.53-1.18 (m, 15H), 1.12-1.0 (m, 2H), 1.03
(s, 3H), 0.98-0.75 (m, 12H).
Step 6: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-acetoxy-1-isopropyl-5a,5b,8,
8,11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a--
octadecahydro-3aH-cyclopenta[a]chrysene-3a-carboxylic acid
##STR00015##
[0276] To an ice-cooled solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-formyl-1-isopropyl-5a,5b,8,8,11a-p-
entamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,
13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl acetate (step 5,
33.0 g, 66.465 mmol, 1.0 eq) in t-butanol (330 ml), THF (500 ml)
and 2-methyl 2-butene (60 ml) was added slowly a solution of
NaClO.sub.2 (71.86 g, 797.58 mmol, 12.0 eq) in 385 ml of water
followed by NaH.sub.2PO.sub.4 (79.75 g, 664.65 mmol, 10.0 eq) in
390 ml of water (390 ml) over 15 minutes. After stirring at
0.degree. C. for about 10 minutes, the reaction mixture was warmed
to room temperature and stirred for about 1 hour. TLC indicated
starting material was consumed and the desired product was
observed. The reaction mixture was diluted with water (100 ml) and
extracted with ethyl acetate (3.times.500 ml). The combined organic
extracts were dried over sodium sulfate, filtered and evaporated
under reduced pressure. The residue was triturated with n-hexane,
solid formed was collected by filtration and dried under vacuum to
obtain the desired product (33.1 g, yield: 97.3%) as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 4.49 (dd, J=10.2,
5.7 Hz, 1H), 3.27-3.18 (m, 1H), 2.78-2.71 (m, 1H), 2.58 (d, J=18.9
Hz, 1H), 2.50-2.43 (m, 1H), 2.19 (d, J=18.6 Hz, 1H), 2.05 (s, 3H),
2.02-1.82 (m, 3H), 1.81-1.51 (m, 5H), 1.51-1.25 (m, 6H), 1.22 (d,
J=1.5 Hz, 3H), 1.20 (d, J=1.5 Hz, 3H), 1.17-1.09 (m, 1H), 1.05 (s,
3H), 1.03-0.98 (m, 1H), 0.94 (s, 3H), 0.91 (s, 3H), 0.85 (s, 3H),
0.84 (s, 3H), 0.80 (m, 1H); ESI-MS: m/z 535.42 (M+Na).sup.+.
Step 7: Synthesis of
(3aR,5aR,5bR,7aR,9,11aR,11bR,13aS)-1-isopropyl-3a-((((4-methoxy
benzyl)oxy)carbonyl)amino)-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b-
,6,7,7a,8,9,10,
11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl
acetate
##STR00016##
[0278] To a stirred solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-acetoxy-1-isopropyl-5a,5b,8,8,11a-p-
entamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadeca-
hydro-3aH-cyclopenta[a]chrysene-3a-carboxylic acid (step 6, 33.0 g,
64.45 mmol, 1.0 eq) in 1,2-dichloroethane (500 ml) was added
triethyl amine (22.43 ml, 161.13 mmol, 2.5 eq), followed by
diphenylphosphonic azide (18.0 ml, 83.78 mmol, 1.3 eq). After 15
minutes stirring at room temperature, the solution was heated to
reflux for about 100 minutes. After which it was converted
completely to the isocyanate by TLC, p-methoxybenzyl alcohol (9.9
ml, 83.78 mmol, 1.3 eq) was added and reflux was continued for
about 4 hours. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was evaporated
under reduced pressure and the residue was purified by silicagel
column chromatography by using 0-3% methanol in dichloromethane
gradient. The fractions containing the product were combined and
concentrated under reduced pressure to give the desired product
(38.0 g, yield: 91.1%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 7.30 (d, J=8.7 Hz, 2H), 6.87 (d, J=8.7 Hz,
2H), 4.98 (s, 2H), 4.83 (s, 1H), 4.48 (dd, J=10.5, 6.0 Hz, 1H),
3.80 (s, 3H), 3.22-3.0 (m, 1H), 2.82-2.58 (m, 2H), 2.55-2.20 (m,
2H), 2.05 (s, 3H), 1.98-1.60 (m, 7H), 1.58-1.30 (m, 7H), 1.28-1.12
(m, 8H), 1.07 (s, 3H), 0.92 (m, 6H), 0.85-0.78 (m, 7H); ESI-MS: m/z
670.51 (M+Na).sup.+.
Step 8: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-amino-1-isopropyl-5a,5b,8,8,
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-o-
ctadecahydro-2H-cyclopenta[a]chrysen-9-yl acetate
##STR00017##
[0280] To a stirred solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-3a-((((4-methoxybenzyl)ox-
y)carbonyl)amino)-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl
acetate (step 7, 38.0 g, 58.73 mmol, 1.0 eq) in DCM (320 ml) at
0.degree. C. was added TFA (80 ml). The reaction mixture was
allowed to stir at room temperature for overnight. TLC indicated
starting material was consumed and the desired product was
observed. The reaction mixture was evaporated under reduced
pressure, water (100 ml) was added, cooled to 0.degree. C., pH
adjusted to 8.0 with saturated NaHCO.sub.3 solution and extracted
with DCM (3.times.600 ml). The combined organic extracts were dried
over sodium sulfate, filtered and evaporated under reduced
pressure. The residue was purified by silicagel column
chromatography by using a 0-3% methanol in dichloromethane
gradient. The fractions containing the product were combined and
concentrated under reduced pressure to obtain the desired product
(26.0 g, yield: 91.87%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 4.48 (m, 1H), 3.20-3.05 (m, 1H), 2.33 (d,
J=18.6 Hz, 1H), 2.23 (d, J=18.6 Hz, 1H), 2.05 (s, 3H), 1.98-1.72
(m, 4H), 1.72-1.50 (m, 7H), 1.46-1.26 (m, 5H), 1.26-1.0 (m, 11H),
0.93 (s, 3H), 0.91 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H), 0.80 (m,
1H).
Step 9: Synthesis of
2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid
##STR00018##
[0281] Method 1:
[0282] To a stirred solution of 2-Amino-2-methylpropanoic acid (30
g, 290.92 mmol, 1.0 eq) in 1,4-dioxane (300 ml) at 0.degree. C. was
added 2N NaOH solution (300 ml) followed by (Boc).sub.2O (95.13 g,
436.38 mmol, 1.5 eq). The reaction mixture was allowed to stir at
room temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The organic phase
was evaporated under reduced pressure, the reaction mixture was
diluted with water (50 ml), cooled to 0.degree. C., pH adjusted to
5 with 1N HCl and then extracted with DCM (3.times.300 ml). The
combined organic extracts were washed with water (300 ml), brine
(100 ml) solution, dried over Na.sub.2SO.sub.4, filtered and
evaporated under reduced pressure. The residue was stirred with
n-hexane (300 ml) at room temperature for about 30 minutes. The
obtained solid was filtered and dried under vacuum to obtain the
desired product (38.0 g, yield: 64.4%) as a white solid.
Method 2:
[0283] To a stirred solution of 2-carboxypropan-2-aminium chloride
(15.0 g, 145.63 mmol) in 1,4-dioxane (75 mL), added 2N NaOH
solution (75 mL), (Boc).sub.2O (47.62 g, 218.44 mmol) at 0.degree.
C. and stirred the reaction mixture for about 12 hours at room
temperature. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was washed with
EtOAc (200 mL) to remove the impurities, then aqueous part was
acidified with 1N HCl (pH--2-3) and extracted with CH.sub.2Cl.sub.2
(2.times.200 mL). The combined organic extracts were washed with
water, dried over Na.sub.2SO.sub.4, filtered and evaporated under
reduced pressure. The crude residue was purified by silicagel
column chromatography by using 30% EtOAc: n-Hexane as an eluent to
afford the desired product (15.0 g, yield: 50.74%) as an off white
solid. .sup.1H NMR (300 MHz, DMSO): .delta. 12.18 (s, 1H), 7.05 (s,
1H), 1.36 (s, 9H), 1.29 (s, 6H); ES Mass: 226.06 [M+Na].sup.+.
Step 10: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-butoxycarbonyl)
amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl acetate
##STR00019##
[0284] Method 1:
[0285] To a stirred solution of
2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (step 9,
4.539 g, 22.36 mmol, 1.2 eq) in 1,2-DCE (200 ml) was added HATU
(10.62 g, 27.95 mmol, 1.5 eq) followed by DIPEA (19.3 ml, 111.79
mmol, 6.0 eq). The reaction mixture was stirred at room temperature
for about 30 minutes, then (3aR,5aR,5bR,7aR,9S,11aR,11bR, 13
aS)-3a-amino-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,
11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl
acetate (step 8, 9.0 g, 18.63 mmol, 1.0 eq) was added and stirred
at same temperature for overnight. TLC indicated starting material
was consumed and the desired product was observed. The reaction
mixture was evaporated under reduced pressure, diluted with water
(90 ml) and extracted with DCM (3.times.135 ml). The combined
organic extracts were washed with 0.5N HCl (90 ml), water (90 ml)
and brine (45 ml) solution. The organic layer was dried over
Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure.
The residue was purified by silicagel column chromatography by
using 0-2% methanol in dichloromethane gradient. The fractions
containing the product were combined and concentrated under reduced
pressure to give the desired product (11.0 g, yield: 88.7%) as an
off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm
4.91 (brs, 1H), 4.49 (dd, J=10.5, 5.7 Hz, 1H), 3.20-3.08 (m, 1H),
2.86 (m, 1H), 2.68 (d, J=18.6 Hz, 1H), 2.39-2.32 (m, 1H), 2.27 (d,
J=18.6 Hz, 1H), 2.05 (s, 3H), 1.98-1.86 (m, 2H), 1.84 (m, 1H),
1.81-1.52 (m, 6H), 1.49 (s, 3H), 1.47 (s, 3H), 1.42 (s, 9H),
1.39-1.34 (m, 3H), 1.29-1.19 (m, 7H), 1.18-1.13 (m, 1H), 1.17 (s,
3H), 1.10-1.01 (m, 2H), 0.92 (s, 3H), 0.91 (s, 3H), 0.856 (s, 3H),
0.85 (s, 3H), 0.80 (m, 1H); ESI-MS: m/z 691.5 (M+Na).sup.+.
Method 2:
[0286] To a stirred solution of
2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (step 9, 7.56
g, 37.21 mmol, 1.2 eq) in DMF (150 ml) was added EDCI (9.24 g 48.3
mmol 1.5 eq) followed by DMAP (11.8 g 96.77 mmol 3 eq) at 0.degree.
C. The reaction mixture was stirred at room temperature for about
30 minutes, then (3aR,5aR,5bR,7aR,9S,11aR,11bR, 13
aS)-3a-amino-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,
11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl
acetate (step 8, 15.0 g, 31.0 mmol, 1.0 eq) was added and stirred
at room temperature for about 4 hours. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was diluted with ice water (450 ml) and white
solid was obtained. The solid was filtered, dissolved in DCM,
washed with water and brine solution. The organic layer was dried
over Na.sub.2SO.sub.4, evaporated under reduced pressure to give
the desired product (17.5 g, yield: 80%) as an off-white solid.
Step 11: Synthesis of tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-3aH-cyclopenta[a]chrysen-3a-yl)amino)-2-methyl-1-oxopropan-2-yl-
) carbamate
##STR00020##
[0288] To a stirred solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-butoxy
carbonyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethy-
l-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl acetate (step 10, 11.0 g, 16.45 mmol, 1.0 eq) in
MeOH (110 ml), THF (55 ml) and water (28 ml) at 0.degree. C. was
added NaOH (6.582 g, 164.57 mmol, 10.0 eq). The mixture was removed
from the ice bath and was stirred at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The organic phase was evaporated
under reduced pressure, the reaction mixture was diluted with water
(165 ml) and extracted with DCM (3.times.165 ml). The combined
organic extracts were washed with water (110 ml), brine solution
(50 ml), dried over sodium sulfate, filtered and evaporated under
reduced pressure. The residue was purified by silicagel column
chromatography by using 0-3% methanol in dichloromethane gradient.
The fractions containing the product were combined and concentrated
under reduced pressure to give the desired product (10.0 g, yield:
96.92%) as a yellow solid. .sup.1H NMR (300 MHz, CDCl.sub.3):
.delta. ppm 7.0 (brs, 1H), 4.85 (brs, 1H), 3.28-3.08 (m, 2H), 2.86
(m, 1H), 2.68 (d, J=18.6 Hz, 1H), 2.38-2.31 (m, 1H), 2.27 (d,
J=18.6 Hz, 1H), 1.97-1.73 (m, 4H), 1.70-1.65 (m, 3H), 1.64-1.53 (m,
3H), 1.49 (s, 3H), 1.46 (s, 3H), 1.42 (s, 9H), 1.40-1.25 (m, 5H),
1.25 (s, 3H), 1.22 (s, 3H), 1.17 (s, 3H), 1.10-1.02 (m, 1H), 0.97
(s, 3H), 0.94 (s, 3H), 0.89 (s, 3H), 0.77 (s, 3H), 0.75-0.68 (m,
1H); ESI-MS: m/z 649.5 (M+Na).sup.+.
Step 12: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-butoxy
carbonyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethy-
l-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00021##
[0289] Method 1:
[0290] To a stirred solution of tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,
11b,12,13,13a-octadecahydro-3
aH-cyclopenta[a]chrysen-3a-yl)amino)-2-methyl-1-oxo
propan-2-yl)carbamate (step 11, 10.0 g, 15.95 mmol, 1.0 eq) in DCM
(100 ml) at 0.degree. C. under nitrogen atmosphere was added
Et.sub.3N (11.12 ml, 79.75 mmol, 5.0 eq), DMAP (0.969 g, 7.975
mmol, 0.5 eq),
(1S,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (prepared as described in WO 2011/007230 A2, 6.27 g, 23.92
mmol, 1.5 eq) and 2,4,6-trichlorobenzoyl chloride (4.98 ml, 31.89
mmol, 2.0 eq). The mixture was removed from the ice bath and was
stirred at room temperature for overnight. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was evaporated under reduced pressure, diluted
with water (100 ml) and extracted with DCM (3.times.100 ml). The
combined organic extracts were washed with 0.5N HCl (100 ml), water
(100 ml), brine solution (50 ml), dried over sodium sulfate,
filtered and evaporated under reduced pressure. The residue was
purified by silicagel column chromatography by using 0-10% MeOH in
DCM gradient. The fractions containing the product were combined
and concentrated under reduced pressure to give the desired product
(10.0 g, yield: 71.9%) as an off-white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 7.38-7.32 (m, 5H), 6.96 (brs, 1H), 5.15,
5.09 (ABq, J.sub.AB=12.3 Hz, 2H), 4.84 (brs, 1H), 4.45 (dd, J=11.1,
4.5 Hz, 1H), 3.20-3.08 (m, 1H), 2.88-2.58 (m, 5H), 2.40-2.33 (m,
1H), 2.27 (d, J=18.6 Hz, 1H), 2.09-2.02 (m, 1H), 2.0-1.82 (m, 3H),
1.81-1.70 (m, 3H), 1.65-1.54 (m, 4H), 1.49 (s, 3H), 1.46 (s, 3H),
1.42 (s, 9H), 1.40-1.30 (m, 3H), 1.34 (s, 3H), 1.30-1.17 (m, 8H),
1.14 (s, 3H), 1.11-1.0 (m, 1H), 0.97 (s, 3H), 0.93 (s, 3H), 0.91
(s, 3H), 0.86 (s, 3H), 0.85 (s, 3H), 0.79 (m, 1H).
Method 2:
[0291] To a stirred solution of
(1S,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclo
butane-1-carboxylic acid (prepared as described in WO 2011/007230
A2, 0.54 g, 3.89 mmol, 1.5 eq) in DMF (20 ml) at 0.degree. C. under
nitrogen atmosphere was added EDCI (0.99 g, 5.18 mmol, 2 eq), HOBT
(0.52 g, 3.89 mmol, 1.5 eq), DMAP (0.15 g, 1.29 mmol, 0.5 eq),
added Triethylamine (1.08 ml 7.78 mmol, 3 eq) and stirred it for
about 30 minutes. Then added tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-3aH-cyclopenta[a]chrysen-3a-yl)amino)-2-methyl-1-oxopropan-2-yl-
)carbamate (step 11, 2.0 g, 2.59 mmol, 1 eq). The reaction mixture
was stirred at room temperature for about 14 hours. TLC indicated
starting material was consumed and the desired product was
observed. The reaction mixture was quenched with ice cold water
then filtered through Buchner funnel. The solid was separated, then
the solid compound was dissolved in DCM, washed with sodium
bicarbonate, water, brine solution then dried over sodium sulfate
and concentrated under reduced pressure to give a crude compound.
The crude compound was purified by flash silica column
chromatography by using 100-200 silica gel, then the product was
eluted at 2% MeOH in DCM (1.5 g, yield: 65.21%) as an off-white
solid.
Step 13: Synthesis of
1-((3aR,5aR,5bR,5bR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methyl
propanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,
11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-di methylcyclobutane-1,3-dicarboxylate
[0292] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-butoxycarbonyl)amino)-
-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5-
,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step
12, 10.0 g, 11.47 mmol, 1.0 eq) in DCM (80 ml) was added
trifluoroacetic acid (20 ml). The reaction mixture was stirred at
room temperature for about 14 hours. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was evaporated under reduced pressure, cooled to
0.degree. C., pH adjusted to 8.0 with saturated sodium bicarbonate
solution and extracted with DCM (3.times.100 ml). The combined
organic extracts were dried over sodium sulfate, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-5% methanol in
dichloromethane gradient. The fractions containing the product were
combined and concentrated under reduced pressure to give the
desired product (8.0 g, yield: 90.4%) as an off-white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.99 (brs, 1H),
7.38-7.32 (m, 5H), 5.15, 5.09 (ABq, J.sub.AB=12.3 Hz, 2H), 4.45
(dd, J=11.1, 4.5 Hz, 1H), 3.20-3.09 (m, 1H), 2.89-2.61 (m, 5H),
2.43-2.35 (m, 1H), 2.30 (d, J=18.6 Hz, 1H), 2.10-1.92 (m, 3H),
1.90-1.65 (m, 4H), 1.53-1.40 (m, 4H), 1.40-1.30 (m, 12H), 1.27-1.15
(m, 8H), 1.12 (s, 3H), 1.09-1.01 (m, 1H), 0.96 (s, 3H), 0.94 (s,
3H), 0.91 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H), 0.79 (m, 1H);
ESI-MS: m/z 771.6 (M+H).sup.+.
Intermediate 2: Preparation of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00022##
[0293] Step 1: Synthesis of
(1R,3S)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
2,4,6-trichlorobenzoic anhydride
##STR00023##
[0295] To a stirred solution of
(1R,3S)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (prepared as described in WO 2014/105926 A1, 1.0 g, 3.812
mmol, 1.0 eq) in THF (10 ml) at 0.degree. C. under nitrogen
atmosphere was added triethylamine (1.59 ml, 11.436 mmol, 3.0 eq)
followed by 2,4,6-trichlorobenzoyl chloride (0.71 ml, 4.574 mmol,
1.2 eq). The reaction mixture was allowed to stir at room
temperature for about 4 hours. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was evaporated under reduced pressure to obtain the desired product
(1.8 g) as an oil, which is used as such for next step without
further purification.
Step 2. Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9,11aR,11bR,13aS)-3a-(2-((tert-butoxy
carbonyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,
8,11a-pentamethyl-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl)
(1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00024##
[0297] To a stirred solution of tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,
11b,12,13,13a-octadecahydro-3
aH-cyclopenta[a]chrysen-3a-yl)amino)-2-methyl-1-oxo
propan-2-yl)carbamate (Intermediate 1-step 11, 1.5 g, 2.392 mmol,
1.0 eq) in Toluene (15 ml) at 0.degree. C. under nitrogen
atmosphere was added DMAP (0.730 g, 5.981 mmol, 2.5 eq) and
(1R,3S)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
2,4,6-trichloro benzoic anhydride (step 1, 1.685 g, 3.588 mmol, 1.5
eq) dissolved in toluene (15 ml). The reaction mixture was allowed
to stir at room temperature for about 30 minutes and then heated to
reflux for overnight. TLC indicated starting material was consumed
and the desired product was observed. The reaction mixture was
evaporated under reduced pressure, diluted with water (20 ml) and
extracted with DCM (3.times.30 ml). The combined organic extracts
were washed with water (20 ml), brine solution (20 ml), dried over
sodium sulfate, filtered and evaporated under reduced pressure. The
residue was purified by silicagel column chromatography by using
0-3% methanol in dichloromethane gradient. The fractions containing
the expected product were combined and concentrated under reduced
pressure to obtain the desired product (1.0 g, yield: 48.07%) as a
white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.35
(m, 5H), 5.15, 5.10 (ABq, J.sub.AB=12.3 Hz, 2H), 4.46 (dd, J=11.1,
4.5 Hz, 1H), 3.20-3.08 (m, 1H), 3.0-2.58 (m, 5H), 2.40-2.23 (m,
2H), 2.12-2.02 (m, 1H), 1.98-1.82 (m, 2H), 1.80-1.67 (m, 2H),
1.65-1.53 (m, 7H), 1.50 (s, 3H), 1.47 (s, 3H), 1.43 (s, 9H),
1.40-1.30 (m, 4H), 1.33 (s, 3H), 1.24 (d, J=6.9 Hz, 3H), 1.21 (d,
J=6.9 Hz, 3H), 1.14 (s, 3H), 1.08-1.0 (m, 1H), 0.97-0.90 (m, 9H),
0.87-0.78 (m, 7H); ESI-MS: m/z 893.54 (M+Na).sup.+.
Step 3: Synthesis of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropan
amido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,-
8,9,10,11,11a,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl) 3-benzyl
(1R,3S)-2,2-dimethyl cyclobutane-1,3-dicarboxylate
[0298] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-butoxycarbonyl)amino)-
-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5-
,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step
2, 1.0 g, 1.148 mmol, 1.0 eq) in DCM (8 ml) was added
trifluoroacetic acid (2 ml). The reaction mixture was stirred at
room temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was evaporated under reduced pressure, diluted with water (10 ml),
cooled to 0.degree. C., pH adjusted to 8.0 with saturated sodium
bicarbonate solution and extracted with DCM (3.times.50 ml). The
combined organic extracts were washed with water (30 ml), dried
over sodium sulfate, filtered and evaporated under reduced
pressure. The residue was purified by silicagel column
chromatography by using 0-5% methanol in dichloromethane gradient.
The fractions containing the expected product were combined and
concentrated under reduced pressure to give the desired product
(0.8 g, yield: 90.3%) as a white solid.
Intermediate 3: Preparation of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclo
propane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4-
,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00025##
[0299] Step 1: Synthesis of
1-((tert-butoxycarbonyl)amino)cyclopropane-1-carboxylic acid
##STR00026##
[0301] To a stirred solution of 1-aminocyclopropane-1-carboxylic
acid (30 g, 149.25 mmol, 1.0 eq) in 1,4-dioxane (300 ml) at
0.degree. C. was added 2N NaOH solution (300 ml) followed by
(Boc).sub.2O (48.80 gr, 223.88 mmol, 1.5 eq). The reaction mixture
was allowed to stir at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (50 ml),
cooled to 0.degree. C., pH adjusted to 5 with 1N HCl and then
extracted with DCM (3.times.300 ml). The combined organic extracts
were washed with water (300 ml), brine (100 ml) solution, dried
over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure. The residue was stirred with n-hexane (300 ml) at room
temperature for about 30 minutes, the obtained solid was filtered
and dried under vacuum to obtain the desired product (32.0 g,
yield: 53.60%) as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 12.26 (brs, 1H), 7.40 (s, 1H), 1.36 (s,
9H), 1.25 (d, J=12 Hz, 2H), 0.95-0.91 (d, J=12 Hz, 2H); ESI-MS: m/z
226.43 (M+Na).sup.+.
Step 2: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)
amino)cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl acetate
##STR00027##
[0303] To a stirred solution of
1-((tert-butoxycarbonyl)amino)cyclopropane-1-carboxylic acid (step
1, 12.48 g, 62.08 mmol, 1.5 eq) in DMF (150 ml) at 0.degree. C.
under nitrogen atmosphere was added EDCI (15.90 g, 82.81 mmol, 2
eq), HOBT (8.38 g, 62.11 mmol, 1.5 eq), DMAP (2.52 g, 20.70 mmol,
0.5 eq), and added Triethylamine (17.28 ml, 124.22 mmol, 3 eq) and
stirred it for about 30 minutes. Then added
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-amino-1-isopropyl-5a,5b,8,8,11a-pe-
ntamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysene-9-yl acetate
(Intermediate 1-step 8, 20.0 g, 41.40 mmol, 1 eq) and the reaction
mixture was stirred at room temperature for about 14 hours. TLC
indicated starting material was consumed and the desired product
was observed. The reaction mixture was quenched with ice cold water
then filtered through Buchner funnel then solid was separated, then
that solid compound was dissolved in DCM and washed with sodium
bicarbonate, water, brine solution then dried over sodium sulfate
and concentrated under reduced pressure to give a crude compound.
The crude compound was purified by flash silica column
chromatography (100-200 silica gel) using 1.5% MeOH in DCM as an
eluent to obtain the desired product (20.0 g, yield: 72.72%) as an
off pale yellow colour solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 7.37-7.20 (m, 1H), 7.02-6.81 (m, 1H), 4.41-4.36 (m,
1H), 4.06-3.99 (m, 1H), 3.74-3.47 (m, 1H), 3.23-3.10 (m, 1H),
2.97-2.89 (m, 1H), 2.80-2.73 (m, 1H), 2.38-2.27 (m, 2H), 2.20-2.14
(m, 2H), 1.90 (m, 3H), 1.86-1.49 (m, 8H), 1.39-1.34 (m, 9H),
1.28-1.17 (m, 3H), 1.15-1.09 (m, 9H), 0.99-0.94 (m, 3H), 0.89-0.87
(m, 6H), 0.83-0.80 (s, 9H); ESI-MS: m/z 689.43 (M+Na).sup.+.
Step 3: Synthesis of tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,
13a-octadecahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclopropyl)ca-
rbamate
##STR00028##
[0305] To a stirred solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pent-
amethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclopenta[a]chrysen-9-yl acetate (step 2, 20 g, 30.03 mmol,
1.0 eq) in MeOH (200 ml), THF (100 ml) and water (50 ml) at
0.degree. C. was added NaOH (12.01 g, 300.30 mmol, 10.0 eq). The
mixture was removed from the ice bath and was stirred at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The organic phase
was evaporated under reduced pressure, the reaction mixture was
diluted with water (180 ml) and extracted with DCM (3.times.180
ml). The combined organic extracts were washed with water (100 ml),
brine solution (50 ml), dried over sodium sulfate, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-3% methanol in
dichloromethane gradient. The fractions containing the product were
combined and concentrated under reduced pressure to give the
desired product (16.0 g, yield: 85.5%) as a white solid. .sup.1H
NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 8.32 (m, 1H), 7.20 (m,
1H), 4.32-4.31 (m, 1H), 3.72-3.67 (m, 1H), 3.12-2.75 (m, 3H),
2.38-2.09 (m, 4H), 1.85 (m, 4H), 1.65-1.23 (m, 19H), 1.13-1.08 (m,
10H), 0.95-0.83 (m, 13H), 0.66 (s, 3H); ESI-MS: m/z 647.43
(M+Na).sup.+.
Step 4: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pent-
amethyl-2-oxo-3,3a,4,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00029##
[0307] To a stirred solution of
(1S,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (prepared as described in WO 2011/007230 A2, 10.07 g, 38.46
mmol, 1.5 eq) in DMF (120 ml) at 0.degree. C. under nitrogen
atmosphere was added EDCI (9.84 g, 51.28 mmol, 2 eq), HOBT (5.19 g,
38.46 mmol, 1.5 eq), DMAP (1.56 g, 12.82 mmol, 0.5 eq) and added
Triethylamine (10.70 ml, 76.92 mmol, 3 eq) and stirred it for about
30 minutes. Then added tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclopropyl)carbamate
(step 3, 16.0 g, 25.64 mmol, 1 eq) and the reaction mixture was
stirred at room temperature for about 14 hours. TLC indicated
starting material was consumed and the desired product was
observed. The reaction mixture was quenching with ice cold water
then filtered through Buchner funnel then solid was separated, then
that solid compound was dissolved in DCM, washed with sodium
bicarbonate, water and brine solution then dried over sodium
sulfate and concentrated under reduced pressure to give a crude
compound. The crude compound was purified by flash silica column
chromatography (100-200 silica gel) using 2% MeOH in DCM gradient.
The fractions containing the product were combined and concentrated
under reduced pressure to give the desired product (18.0 g, yield:
80.89%) as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 7.36-7.34 (m, 5H), 7.32-7.20 (m, 2H), 5.14, 5.08 (ABq,
J.sub.AB=12.3 Hz, 2H), 4.35 (m, 1H), 3.74-3.69 (m, 2H), 3.12-2.84
(m, 5H), 2.38-2.14 (m, 4H), 1.96-1.90 (m, 3H), 1.73-1.1.58 (m, 8H),
1.39-1.36 (m, 14H), 1.13-1.09 (m, 14H), 0.89-0.81 (m, 16H); ESI-MS:
m/z 891.32 (M+Na).sup.+.
Step 5: Synthesis of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclopropane-1-carboxam-
ido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9-
,10,11, ha,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl) 3-benzyl
(1S,3R)-2,2-di methylcyclobutane-1,3-dicarboxylate
[0308] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)amino)-
cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,-
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopent-
a[a]chrysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(step 4, 18 g, 20.73 mmol, 1.0 eq) in DCM (160 ml) was added
trifluoroacetic acid (36 ml). The reaction mixture was stirred at
room temperature for about 3 hours. TLC indicated starting material
was consumed and the desired product was observed. The reaction
mixture was slowly poured in to cold sodium bicarbonate solution,
pH adjusted to 8.0 then filtered through celite pad and the
filtrate was extracted with DCM (3.times.200 ml). The combined
organic layer was washed with water (200 ml), dried over sodium
sulfate, and evaporated under reduced pressure to give the desired
product (13.0 g, yield: 81.76%) as an off-white solid. .sup.1H NMR
(300 MHz, DMSO-d.sub.6): .delta. ppm 7.36-7.34 (m, 5H), 7.32-7.20
(m, 2H), 5.14, 5.08 (ABq, J.sub.AB=12.3 Hz, 2H), 4.35 (m, 1H),
3.74-3.69 (m, 2H), 3.12-2.84 (m, 5H), 2.38-2.14 (m, 4H), 1.96-1.90
(m, 3H), 1.73-1.58 (m, 7H), 1.39-1.36 (m, 7H), 1.13-1.09 (m, 14H),
0.89-0.81 (m, 16H); ESI-MS: m/z 791.52 (M+Na).sup.+.
Intermediate 4: Preparation of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclo
propane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4-
,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1R,3 S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00030##
[0309] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pent-
amethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclopenta[a]chrysen-9-yl)
(1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00031##
[0311] To a stirred solution of
(1R,3S)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (prepared as described in WO 2014/105926 A1, 10.11 g, 38.46
mmol, 1.5 eq) in DMF (120 ml) at 0.degree. C. under nitrogen
atmosphere was added EDCI (14.76 g, 76.92 mmol, 2 eq), HOBT (7.78
g, 57.66 mmol, 1.5 eq), DMAP (2.3 g, 19.22 mmol, 0.5 eq) and added
Triethylamine (15.53 ml, 115.32 mmol, 3 eq) and stirred it for
about 30 minutes. Then added tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclopropyl)carbamate
(Intermediate 3-step 3, 16.0 g, 25.64 mmol, 1 eq). The reaction
mixture was stirred at room temperature for about 14 hours. TLC
indicated starting material was consumed and the desired product
was observed. The reaction mixture was quenched with ice cold water
then filtered through Buchner funnel then solid was separated, then
that solid compound was dissolved in DCM and washed with sodium
bicarbonate, water and brine solution then dried over sodium
sulfate and concentrated under reduced pressure to give a crude
compound. The crude compound was purified by flash silica column
chromatography (100-200 silica gel) using 2% MeOH in DCM gradient.
The fractions containing the product were combined and concentrated
under reduced pressure to give the desired product (18.0 g, yield:
80.89%) as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 8.32 (m, 1H), 7.20 (m, 1H), 4.32-4.31 (m, 1H),
3.72-3.67 (m, 1H), 3.12-2.75 (m, 3H), 2.38-2.09 (m, 4H), 1.85 (m,
4H), 1.65-1.23 (m, 19H), 1.13-1.08 (m, 10H), 0.95-0.83 (m, 13H),
0.66 (s, 3H); ESI-MS: m/z 647.43 (M+Na).sup.+.
Step 2: Synthesis of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclopropane-1-carboxam-
ido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9-
,10,11,
11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1R,3S)-2,2-di methylcyclobutane-1,3-dicarboxylate
[0312] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)amino)-
cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 1, 18 g,
20.73 mmol, 1.0 eq) in DCM (160 ml) was added trifluoroacetic acid
(36 ml). The reaction mixture was stirred at room temperature for
about 3 hours. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was slowly
poured in to cold sodium bicarbonate solution, pH adjusted to 8.0
then filtered through celite pad, the filtrate was extracted with
DCM (3.times.200 ml). The combined organic layer was washed with
water (200 ml), dried over sodium sulfate, and evaporated under
reduced pressure give the desired product (13.0 g, yield: 81.76%)
as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta.
ppm 7.36-7.34 (m, 5H), 7.32-7.20 (m, 2H), 5.14, 5.08 (ABq,
J.sub.AB=12.3 Hz, 2H), 4.35 (m, 1H), 3.74-3.69 (m, 2H), 3.12-2.84
(m, 5H), 2.38-2.14 (m, 4H), 1.96-1.90 (m, 3H), 1.73-1.58 (m, 7H),
1.39-1.36 (m, 7H), 1.13-1.09 (m, 14H), 0.89-0.81 (m, 16H); ESI-MS:
m/z 791.52 (M+Na).sup.+.
Intermediate 5: Preparation of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclo
butane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,-
5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00032##
[0313] Step 1: Synthesis of
1-((tert-butoxycarbonyl)amino)cyclobutane-1-carboxylic acid
##STR00033##
[0315] To a stirred solution of 1-aminocyclobutane-1-carboxylic
acid (30 g, 139.53 mmol, 1.0 eq) in 1,4-dioxane (300 ml) at
0.degree. C. was added 2N NaOH solution (300 ml) followed by
(Boc).sub.2O (45.62 g, 209.30 mmol, 1.5 eq). The reaction mixture
was allowed to stir at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (50 ml),
cooled to 0.degree. C., pH adjusted to 5 with 1N HCl and then
extracted with DCM (3.times.300 ml). The combined organic extracts
were washed with water (300 ml), brine (100 ml) solution, dried
over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure. The residue was stirred with n-hexane (300 ml) at room
temperature for about 30 minutes, the obtained solid was filtered
and dried under vacuum to give the desired product (35.0 g, yield:
62.41%) as a white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 12.19 (brs, 1H), 7.70-7.16 (m, 1H), 2.45-2.36 (m, 2H),
2.12-2.03 (m, 2H), 1.86-1.78 (m, 2H), 1.36 (brs, 9H); ESI-MS: m/z
238.13 (M+Na).sup.+.
Step 2: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)
amino)cyclobutane-1-carboxamido)-1-isopropyl-5a,5b,8,
8,11a-pentamethyl-2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl acetate
##STR00034##
[0317] To a stirred solution of
1-((tert-butoxycarbonyl)amino)cyclobutane-1-carboxylic acid (step
1, 13.35 g, 62.11 mmol, 1.5 eq) in DMF (160 ml) at 0.degree. C.
under nitrogen atmosphere was added EDCI (15.90 g, 82.81 mmol, 2
eq), HOBT (8.38 g, 62.11 mmol, 1.5 eq), DMAP (2.52 g, 20.70 mmol,
0.5 eq), and added Triethylamine (17.28 ml 124.22 mmol, 3 eq) and
stirred it for about 30 minutes. Then added
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-amino-1-isopropyl-5a,5b,8,8,11a-pe-
ntamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,
12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl acetate
(Intermediate 1-step 8, 20.0 g, 41.40 mmol, 1 eq) and the reaction
mixture was stirred at room temperature for about 14 hours. TLC
indicated starting material was consumed and the desired product
was observed. The reaction mixture was quenched with ice cold water
then filtered through Buchner funnel then solid was separated, then
that solid compound was dissolved in DCM and washed with sodium
bicarbonate, water and brine solution then dried over sodium
sulfate and concentrated under reduced pressure to give a crude
compound. The crude compound was purified by flash silica column
chromatography (100-200 silica gel) using 2% MeOH in DCM gradient.
The fractions containing the product were combined and concentrated
under reduced pressure to give the desired product (22.0 g, yield:
78.15%) as an off-pale yellow solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 7.37-7.20 (m, 1H), 7.02-6.81 (m, 1H),
4.40-4.35 (m, 1H), 4.06-3.98 (m, 1H), 3.73-3.49 (m, 1H), 3.25-3.14
(m, 1H), 2.97-2.90 (m, 1H), 2.82-2.74 (m, 1H), 2.45-2.36 (m, 2H),
2.31-2.26 (m, 2H), 2.16-2.10 (m, 2H), 1.92 (m, 3H), 1.86-1.43 (m,
8H), 1.38-1.33 (m, 9H), 1.27-1.18 (m, 3H), 1.15-1.11 (m, 9H),
0.99-0.95 (m, 3H), 0.90-0.86 (m, 6H), 0.84-0.81 (s, 9H); ESI-MS:
m/z 703.43 (M+Na).sup.+.
Step 3: Synthesis of tert-butyl
(1-(((3aR,5R,5aR,75bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,-
8,8,11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-
-octadecahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclobutyl)carbama-
te
##STR00035##
[0319] To a stirred solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclobutane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-penta-
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahyd-
ro-2H-cyclopenta[a]chrysen-9-yl acetate (step 2, 22 g, 32.35 mmol,
1.0 eq) in MeOH (180 ml), THF (90 ml) and water (50 ml) at
0.degree. C. was added NaOH (12.94 g, 323.52 mmol, 10.0 eq). The
mixture was removed from the ice bath and was stirred at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The organic phase
was evaporated under reduced pressure, the reaction mixture was
diluted with water (180 ml) and extracted with DCM (3.times.180
ml). The combined organic extracts were washed with water (100 ml),
brine solution (50 ml), dried over sodium sulfate, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-3% methanol in
dichloromethane gradient. The fractions containing the product were
combined and concentrated under reduced pressure to give the
desired product (18 g, yield: 87.20%) as a white solid. .sup.1H NMR
(300 MHz, DMSO-d.sub.6): .delta. ppm 8.35 (m, 1H), 7.22 (m, 1H),
4.34-4.30 (m, 1H), 3.75-3.68 (m, 1H), 3.14-2.77 (m, 3H), 2.52-2.43
(m, 2H), 2.40-2.11 (m, 4H), 1.89 (m, 4H), 1.69-1.33 (m, 19H),
1.15-1.11 (m, 10H), 0.99-0.85 (m, 13H), 0.68 (s, 3H); ESI-MS: m/z
661.42 (M+Na).sup.+.
Step 4: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclobutane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-penta-
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahyd-
ro-2H-cyclopenta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00036##
[0321] To a stirred solution of
(1S,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (prepared as described in WO 2011/007230 A2, 11.08 g, 42.31
mmol, 1.5 eq) in DMF (120 ml) at 0.degree. C. under nitrogen
atmosphere was added EDCI (16.25 g, 84.63 mmol, 2 eq), HOBT (5.71
g, 42.31 mmol, 1.5 eq), DMAP (1.72 g, 14.10 mmol, 0.5 eq), and
added triethylamine (11.77 ml, 84.63 mmol, 3 eq) and stirred it for
about 30 minutes. Then added tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclobutyl)carbamate
(step 3, 18 g, 28.21 mmol, 1 eq). The reaction mixture was stirred
at room temperature for about 14 hours. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was quenched with ice cold water then filtered
through Buchner funnel then solid was separated, then that solid
compound was dissolved in DCM and washed with sodium bicarbonate,
water and brine solution then dried over sodium sulfate and
concentrated under reduced pressure to give a crude compound. The
crude compound was purified by flash silica column chromatography
(100-200 silica gel) using 2% MeOH in DCM gradient. The fractions
containing the product were combined and concentrated under reduced
pressure to give the desired product (18 g, yield: 72.58%) as an
off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm
7.37-7.35 (m, 5H), 7.31-7.22 (m, 2H), 5.15, 5.09 (ABq,
J.sub.AB=12.3 Hz, 2H), 4.34-4.30 (m, 1H), 3.77-3.70 (m, 2H),
3.14-2.87 (m, 5H), 2.47-2.38 (m, 2H), 2.32-2.16 (m, 4H), 1.99-1.95
(m, 3H), 1.75-1.60 (m, 8H), 1.38-1.30 (m, 14H), 1.20-1.11 (m, 14H),
0.90-0.83 (m, 16H); ESI-MS: m/z 905.43 (M+Na).sup.+.
Step 5: Synthesis of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclobutane-1-carboxami-
do)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,-
10,11,
11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-di methylcyclobutane-1,3-dicarboxylate
[0322] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)amino)-
cyclobutane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 4, 18 g,
22.95 mmol, 1.0 eq) in DCM (160 ml) was added trifluoroacetic acid
(36 ml). The reaction mixture was stirred at room temperature for
about 3 hours. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was slowly
poured in to cold sodium bicarbonate solution, pH adjusted to 8.0
then filtered through celite pad, the filtrate was extracted with
DCM (3.times.200 ml). The combined organic layer was washed with
water (200 ml), dried over sodium sulfate, and evaporated under
reduced pressure to give the desired product (14 g, yield: 87.77%)
as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta.
ppm 7.34-7.32 (m, 5H), 7.31-7.22 (m, 2H), 5.15, 5.09 (ABq,
J.sub.AB=12.3 Hz, 2H), 4.34-4.32 (m, 1H), 3.77-3.70 (m, 2H),
3.14-2.85 (m, 5H), 2.46-2.41 (m, 2H), 2.38-2.14 (m, 4H), 1.97-1.91
(m, 3H), 1.74-1.60 (m, 7H), 1.40-1.38 (m, 7H), 1.14-1.11 (m, 14H),
0.89-0.83 (m, 16H); ESI-MS: m/z 805.32 (M+Na).sup.+.
Intermediate 6: Preparation of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclo
pentane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4-
,5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00037##
[0323] Step 1: Synthesis of
1-((tert-butoxycarbonyl)amino)cyclopentane-1-carboxylic acid
##STR00038##
[0325] To a stirred solution of 1-aminocyclopentane-1-carboxylic
acid (10 g, 77.51 mmol, 1.0 eq) in 1,4-dioxane (100 ml) at
0.degree. C. was added 2N NaOH solution (100 ml) followed by
(Boc).sub.2O (25.34 g, 116.27 mmol, 1.5 eq). The reaction mixture
was allowed to stir at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (50 ml),
cooled to 0.degree. C., pH adjusted to 5 with 1N HCl and then
extracted with DCM (3.times.300 ml). The combined organic extracts
were washed with water (300 ml), brine (100 ml) solution, dried
over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure. The residue was stirred with n-hexane (300 ml) at room
temperature for about 30 minutes, the obtained solid was filtered
and dried under vacuum to obtain the desired product (10.0 g,
yield: 56.33%) as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 12.11 (s, 1H), 7.10-6.86 (m, 1H),
1.94-1.84 (m, 4H), 1.62-1.58 (m, 4H), 1.36 (s, 9H); ESI-MS: m/z
252.02 (M+Na).sup.+.
Step 2: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)
amino)cyclopentane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-
-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl acetate
##STR00039##
[0327] To a stirred solution of
1-((tert-butoxycarbonyl)amino)cyclopentane-1-carboxylic acid (step
1, 8.5 g, 37.11 mmol, 1.5 eq) in DMF (100 ml) at 0.degree. C. under
nitrogen atmosphere was added EDCI (9.54 g, 49.68 mmol, 2 eq), HOBT
(5.03 g, 37.26 mmol, 1.5 eq), DMAP (1.51 g, 12.42 mmol, 0.5 eq),
and added Triethylamine (10.36 ml 74.53 mmol, 3 eq) and stirred it
for about 30 minutes. Then added
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-amino-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,
13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl acetate
(Intermediate 1-step 8, 12 g, 24.84 mmol, 1 eq). The reaction
mixture was stirred at room temperature for about 14 hours. TLC
indicated starting material was consumed and the desired product
was observed. The reaction mixture was quenched with ice cold water
then filtered through Buchner funnel then solid was separated, then
that solid compound was dissolved in DCM and washed with sodium
bicarbonate, water and brine solution then dried over sodium
sulfate and concentrated under reduced pressure to give a crude
compound. The crude compound was purified by flash silica column
chromatography (100-200 silica gel) using 2% MeOH in DCM gradient.
The fractions containing the product were combined and concentrated
under reduced pressure to give the desired product (10 g, yield:
58.00%) as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 7.10 (brs, 1H), 6.89 (brs, 1H), 4.42-4.36 (m, 1H),
3.74-3.66 (m, 1H), 3.12-3.08 (m, 2H), 2.83-2.79 (m, 1H), 2.35-2.26
(m, 2H), 2.11-1.81 (m, 11H), 1.56-1.47 (m, 9H), 1.35-1.23 (m, 13H),
1.14-1.07 (m, 11H), 0.90-0.80 (m, 10H), 0.76 (brs, 3H); ESI-MS: m/z
717.52 (M+Na).sup.+.
Step 3: Synthesis of tert-butyl
(1-(((3aR,5R,5aR,75bR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,
13a-octadecahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclopentyl)ca-
rbamate
##STR00040##
[0329] To a stirred solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclopentane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pent-
amethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclopenta[a]chrysen-9-yl acetate (step 2, 10 g, 14.40 mmol,
1.0 eq) in MeOH (60 ml), THF (30 ml) and water (15 ml) at 0.degree.
C. was added NaOH (5.76 g, 144.0 mmol, 10.0 eq). The mixture was
removed from the ice bath and was stirred at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The organic phase was evaporated
under reduced pressure, the reaction mixture was diluted with water
(180 ml) and extracted with DCM (3.times.180 ml). The combined
organic extracts were washed with water (100 ml), brine solution
(50 ml), dried over sodium sulfate, filtered and evaporated under
reduced pressure. The residue was purified by silicagel column
chromatography by using 0-3% methanol in dichloromethane gradient.
The fractions containing the product were combined and concentrated
under reduced pressure to give the desired product (7 g, yield:
75.26%) as a white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 7.09 (brs, 1H), 6.89 (brs, 1H), 4.31-4.29 (m, 1H),
3.17-3.15 (m, 1H), 3.11-2.98 (m, 2H), 2.82-2.79 (m, 1H), 2.35-2.27
(m, 2H), 2.11-1.81 (m, 9H), 1.57-1.47 (m, 9H), 1.35-1.23 (m, 12H),
1.19-1.06 (m, 11H), 0.94-0.82 (m, 11H), 0.66 (brs, 3H); ESI-MS: m/z
675.72 (M+Na).sup.+.
Step 4: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclopentane-1-carboxamido)-1-isopropyl-5a,5b,8,8,
a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-2H-cyclopenta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00041##
[0331] To a stirred solution of
(1S,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (prepared as described in WO 2011/007230 A2, 4.2 g, 16.03
mmol, 1.5 eq) in DMF (60 ml) at 0.degree. C. under nitrogen
atmosphere was added EDCI (4.12 g, 21.47 mmol, 2 eq), HOBT (2.17 g,
16.10 mmol, 1.5 eq), DMAP (0.65 g, 5.36 mmol, 0.5 eq), and added
Triethylamine (4.48 ml 32.20 mmol, 3 eq) and stirred it for about
30 minutes. Then added tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclopentyl)carbamate
(step 3, 7 g, 10.73 mmol, 1 eq). The reaction mixture was stirred
at room temperature for about 14 hours. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was quenched with ice cold water then filtered
through Buchner funnel then solid was separated, then that solid
compound was dissolved in DCM and washed with sodium bicarbonate,
water and brine solution then dried over sodium sulfate and
concentrated under reduced pressure to give a crude compound. The
crude compound was purified by flash silica column chromatography
(100-200 silica gel) using 2% MeOH in DCM gradient. The fractions
containing the product were combined and concentrated under reduced
pressure to give the desired product (6 g, yield: 62.41%) as an
off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm
7.37-7.32 (m, 5H), 7.10 (s, 1H), 6.89 (s, 1H), 5.14, 5.08 (ABq,
J.sub.AB=12.3 Hz, 2H), 4.38-4.32 (m, 1H), 3.61-3.59 (m, 1H),
3.12-3.07 (m, 1H), 2.97-2.80 (m, 3H), 2.38-2.31 (m, 4H), 2.22-1.88
(m, 6H), 1.81-1.77 (m, 5H), 1.69-1.57 (m, 8H), 1.35 (m, 10H),
1.26-1.23 (m, 7H), 1.14-1.03 (m, 11H), 0.89-0.78 (m, 14H); ESI-MS:
m/z 919.45 (M+Na).sup.+.
Step 5: Synthesis of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclopentane-1-carboxam-
ido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9-
,10,11, ha,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl) 3-benzyl
(1S,3R)-2,2-di methylcyclobutane-1,3-dicarboxylate
[0332] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)amino)-
cyclopentane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,-
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopent-
a[a]chrysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(step 4, 6.0 g, 6.69 mmol, 1.0 eq) in DCM (60 ml) was added
trifluoroacetic acid (12 ml). The reaction mixture was stirred at
room temperature for about 3 hours. TLC indicated starting material
was consumed and the desired product was observed. The reaction
mixture was slowly poured in to cold sodium bicarbonate solution,
pH adjusted to 8.0 then filtered through celite pad and the
filtrate was extracted with DCM (3.times.200 ml). The combined
organic layer was washed with water (200 ml), dried over sodium
sulfate, and evaporated under reduced pressure to give the desired
product (4.0 g, yield: 75.47%) as an off-white solid. .sup.1H NMR
(300 MHz, DMSO-d.sub.6): .delta. ppm 7.36-7.35 (m, 5H), 7.10 (s,
1H), 5.14, 5.08 (ABq, J.sub.AB=12.3 Hz, 2H), 4.38-4.32 (m, 1H),
3.61-3.59 (m, 1H), 3.12-3.07 (m, 1H), 2.97-2.80 (m, 3H), 2.38-2.31
(m, 4H), 2.22-1.88 (m, 6H), 1.81-1.77 (m, 5H), 1.69-1.35 (m, 11H),
1.26-1.23 (m, 7H), 1.14-1.03 (m, 11H), 0.89-0.78 (m, 14H); ESI-MS:
m/z 819.32 (M+Na).sup.+.
Intermediate 7: Preparation of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclo
hexane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,-
5,5a,5b,6,7,7a,
8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00042##
[0333] Step 1: Synthesis of
1-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid
##STR00043##
[0335] To a stirred solution of 1-aminocyclohexane-1-carboxylic
acid (10 g, 69.93 mmol, 1.0 eq) in 1,4-dioxane (300 ml) at
0.degree. C. was added 2N NaOH solution (100 ml) followed by
(Boc).sub.2O (22.86 g, 104.89 mmol, 1.5 eq). The reaction mixture
was allowed to stir at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (50 ml),
cooled to 0.degree. C., pH adjusted to 5 with 1N HCl and then
extracted with DCM (3.times.300 ml). The combined organic extracts
were washed with water (300 ml), brine (100 ml) solution, dried
over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure. The residue was stirred with n-hexane (300 ml) at room
temperature for about 30 minutes, the obtained solid was filtered
and dried under vacuum to afford the desired product (11.0 g,
yield: 64.74%) as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 12.08 (s, 1H), 6.88 (s, 1H), 1.92-1.88
(m, 2H), 1.62-1.56 (m, 2H), 1.45-1.43 (m, 4H), 1.36 (s, 9H),
1.25-1.19 (m, 2H); ESI-MS: m/z 266.32 (M+Na).sup.+.
Step 2: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)
amino)cyclohexane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2--
oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl acetate
##STR00044##
[0337] To a stirred solution of
1-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (step
1, 9.05 g, 37.26 mmol, 1.5 eq) in DMF (120 ml) at 0.degree. C.
under nitrogen atmosphere was added HBTU (18.83 g, 49.68 mmol, 2
eq), DMAP (1.51 g, 12.42 mmol, 0.5 eq), and added
Diisopropylethylamine (12.81 ml, 74.53 mmol, 3 eq) and stirred it
for about 10 minutes. Then added
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-amino-1-isopropyl-5a,5b,8,8,11a-pe-
ntamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadeca-
hydro-2H-cyclopenta[a]chrysen-9-yl acetate (Intermediate 1-step 8,
12 g, 24.84 mmol, 1 eq). The reaction mixture was stirred at room
temperature for about 14 hours. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was evaporated under reduced pressure, the reaction mixture was
diluted with water (200 ml), and extracted with DCM (3.times.200
ml). The combined organic extracts were washed with water (300 ml),
sodium bicarbonate, and brine solution, then dried over sodium
sulfate, filtered and concentrated under reduced pressure to give a
crude compound, The crude compound was purified by flash silica
column chromatography (100-200 silica gel) using 2% MeOH in DCM as
an eluent. The fractions containing the product were combined and
concentrated under reduced pressure to give the desired product (12
g, yield: 68.22%) as an off-white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 7.04 (brs, 1H), 6.53 (brs, 1H),
4.41-4.37 (m, 1H), 3.69-3.58 (m, 1H), 3.12-3.07 (m, 1H), 2.89-2.82
(m, 1H), 2.73-2.68 (m, 2H), 2.37-2.27 (m, 3H), 2.11-2.05 (m, 2H),
2.02-1.95 (m, 3H), 1.88-1.80 (m, 3H), 1.74-1.71 (m, 6H), 1.60-1.55
(m, 3H), 1.47 (s, 8H), 1.42-1.41 (m, 7H), 1.35-1.23 (m, 3H),
1.39-1.11 (m, 10H), 1.09-1.07 (m, 7H), 1.04-0.80 (m, 5H); ESI-MS:
m/z 731.44 (M+Na).sup.+.
Step 3: Synthesis of tert-butyl
(1-(((3aR,5aR,5bR,7aR,9,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,1-
1a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclohexyl)carbamate
##STR00045##
[0339] To a stirred solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclohexane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-penta-
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahyd-
ro-2H-cyclopenta[a]chrysen-9-yl acetate (step 2, 12 g, 16.94 mmol,
1.0 eq) in MeOH (120 ml), THF (60 ml) and water (30 ml) at
0.degree. C. was added NaOH (6.77 g, 169.4 mmol, 10.0 eq). The
mixture was removed from the ice bath and was stirred at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The organic phase
was evaporated under reduced pressure, the reaction mixture was
diluted with water (180 ml) and extracted with DCM (3.times.180
ml). The combined organic extracts were washed with water (100 ml),
brine solution (50 ml), dried over sodium sulfate, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-3% methanol in
dichloromethane gradient. The fractions containing the product were
combined and concentrated under reduced pressure to give the
desired product (9 g, yield: 79.78%) as a white solid. .sup.1H NMR
(300 MHz, DMSO-d.sub.6): .delta. ppm 7.04 (s, 1H), 6.53 (s, 1H),
4.31-4.30 (m, 1H), 3.11-2.99 (m, 3H), 2.86-2.82 (m, 1H), 2.32-2.28
(m, 2H), 2.11 (m, 1H), 1.89-1.85 (m, 4H), 1.65-1.56 (m, 5H), 1.45
(brs, 9H), 1.35-1.28 (brs, 12H), 1.13-0.99 (m, 14H), 0.94-0.82 (m,
9H), 0.66 (s, 3H); ESI-MS: m/z 689.50 (M+Na).sup.+.
Step 4: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxy
carbonyl)amino)cyclohexane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-penta-
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahyd-
ro-2H-cyclopenta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00046##
[0341] To a stirred solution of
(1S,3R)-3-((benzyloxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid (prepared as described in WO 2011/007230 A2, 5.31 g, 20.27
mmol, 1.5 eq) in DMF (100 ml) at 0.degree. C. under nitrogen
atmosphere was added EDCI (5.18 g, 27.02 mmol, 2 eq), HOBT (2.73 g,
20.27 mmol, 1.5 eq), DMAP (0.82 g, 6.75 mmol, 0.5 eq), and added
Triethylamine (5.63 ml, 40.54 mmol, 3 eq) and stirred it for about
30 minutes. Then added tert-butyl
(1-(((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-9-hydroxy-1-isopropyl-5a,5b,8,8,-
11a-pentamethyl-2-oxo-2,3,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-3aH-cyclopenta[a]chrysen-3a-yl)carbamoyl)cyclohexyl)carbamate
(step 3, 9.0 g, 13.51 mmol, 1 eq). The reaction mixture was stirred
at room temperature for about 14 hours. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was quenched with ice cold water then filtered
through Buchner funnel then solid was separated, then that solid
compound was dissolved in DCM and washed with sodium bicarbonate,
water and brine solution then dried over sodium sulfate and
concentrated under reduced pressure to give a crude compound. The
crude compound was purified by flash silica column chromatography
(100-200 silica gel) using 2% MeOH in DCM as an eluent. The
fractions containing the product were combined and concentrated
under reduced pressure to give the desired product (8 g, yield:
65.09%) as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 8.13 (s, 1H), 7.36-7.35 (m, 5H), 7.34 (s, 1H), 5.14,
5.08 (ABq, J.sub.AB=12.3 Hz, 2H), 4.38-4.32 (m, 1H), 3.13-3.09 (m,
1H), 3.00-2.67 (m, 4H), 2.38-2.25 (m, 5H), 1.96-1.90 (m, 5H),
1.69-1.40 (m, 21H), 1.36 (brs, 9H), 1.15-1.03 (m, 14H), 0.92-0.81
(m, 13H); ESI-MS: m/z 933.63 (M+Na).sup.+.
Step 5: Synthesis of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-aminocyclohexane-1-carboxami-
do)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,-
10,11,
11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
[0342] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-((tert-butoxycarbonyl)amino)-
cyclohexane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 4, 8.0 g,
8.79 mmol, 1.0 eq) in DCM (80 ml) was added trifluoroacetic acid
(16 ml). The reaction mixture was stirred at room temperature for
about 3 hours. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was slowly
poured in to cold sodium bicarbonate solution, pH adjusted to 8.0
then filtered through celite pad, the filtrate was extracted with
DCM (3.times.200 ml). The combined organic layer was washed with
water (200 ml), dried over sodium sulfate, and evaporated under
reduced pressure to give the desired product (6.0 g, yield: 84.26%)
as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta.
ppm 8.13 (s, 1H), 7.36-7.35 (m, 5H), 5.14, 5.08 (ABq, J.sub.AB=12.3
Hz, 2H), 4.38-4.32 (m, 1H), 3.13-3.09 (m, 1H), 3.00-2.67 (m, 4H),
2.38-2.25 (m, 5H), 1.96-1.90 (m, 5H), 1.69-1.23 (m, 23H), 1.15-1.03
(m, 14H), 0.92-0.81 (m, 13H); ESI-MS: m/z 833.55 (M+Na).sup.+.
Intermediate 8: Preparation of
1-(4-chlorophenyl)cyclopropane-1-carboxylic acid
##STR00047##
[0343] Step 1: Synthesis of
1-(4-chlorophenyl)cyclopropane-1-carbonitrile
##STR00048##
[0345] A suspension of 55% sodium hydride (25.27 g, 1052.9 mmol,
5.3 eq) and THF (200 ml) under nitrogen atmosphere was heated to
40.degree. C. and a solution of 2-(4-chlorophenyl) acetonitrile
(30.0 g, 198.67 mmol, 1.0 eq) in THF (50 ml) was added dropwise
over about 30 minutes. The mixture was stirred at 40.degree. C. for
about 30 minutes and a solution of 1,2-dibromoethane (74.3 g,
397.35 mmol, 2.0 eq) in THF (50 ml) was added dropwise over about
30 minutes. The reaction mixture was stirred at 40.degree. C. for
about 1 hour. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was cooled to
0.degree. C., quenched with ice water (250 ml) and extracted with
ethyl acetate (3.times.400 ml). The combined organic extracts were
washed with brine solution (400 ml), dried over sodium sulfate,
filtered and evaporated under reduced pressure to obtain the
desired product (30.0 g) as a semi solid, which is used as such for
next step without further purification. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 7.33 (d, J=8.7 Hz, 2H), 7.24 (d, J=8.7 Hz,
2H), 1.78-1.71 (m, 2H), 1.42-1.35 (m, 2H).
Step 2. Synthesis of 1-(4-chlorophenyl)cyclopropane-1-carboxylic
acid
##STR00049##
[0347] A stirred solution of
1-(4-chlorophenyl)cyclopropane-1-carbonitrile (step 1, 30.0 g,
169.49 mmol, 1.0 eq), sodium hydroxide (20.3 g, 508.47 mmol, 3.0
eq), diethylene glycol (120 ml) and water (35.4 ml) was refluxed
for about 18 hours. TLC indicated starting material was consumed
and the desired product was observed. The reaction mixture was
poured into water (1800 ml) and acidified to pH 4.0 with
concentrated HCl (54 ml). The generated crystals were collected by
filtration and dried under vacuum to obtain the desired product
(30.0 g, yield: 90.3%) as a pale-brown color solid. .sup.1H NMR
(300 MHz, DMSO-d.sub.6): .delta. ppm 12.40 (brs, 1H), 7.34 (m, 4H),
1.48-1.42 (m, 2H), 1.16-1.10 (m, 2H).
Intermediate 9: Preparation of 2,2,2-trifluoroacetate salt of
2-(4-ethylpiperazin-1-yl)acetic acid
##STR00050##
[0348] Step 1: Synthesis of tert-butyl
2-(4-ethylpiperazin-1-yl)acetate
##STR00051##
[0350] To a stirred solution of 1-ethylpiperazine (5.0 g, 43.78
mmol, 1.0 eq) in DCM (100 ml) at 0.degree. C. was added Et.sub.3N
(30.4 ml, 218.93 mmol, 5.0 eq) followed by tert-butyl
2-chloroacetate (9.41 ml, 65.68 mmol, 1.5 eq). The reaction mixture
was stirred at room temperature for overnight. TLC indicated
starting material was consumed and the desired product was
observed. The reaction mixture was diluted with water (150 ml) and
extracted with DCM (3.times.150 ml). The combined organic extracts
were washed with water (100 ml), brine solution (50 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure to
obtain the desired product (9.0 g, yield: 90.09%) as a solid, which
is used as such for next step without further purification. ESI-MS:
m/z 229.09 (M+H).sup.+.
Step 2: Synthesis of 2, 2,2-trifluoroacetate salt of
2-(4-ethylpiperazin-1-yl)acetic acid
[0351] To a stirred solution of tert-butyl
2-(4-ethylpiperazin-1-yl) acetate (step 1, 9.0 g, 39.414 mmol, 1.0
eq) in DCM (72 ml) at 0.degree. C. was added TFA (18 ml). The
reaction mixture was allowed to stir at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was evaporated
under reduced pressure to obtain the desired product (11.2 g) as a
solid, which is used as such for next step without further
purification. ESI-MS: m/z 173.18 (M-TFA+H).sup.+.
Intermediate 10: Preparation of 1-(carboxymethyl)piperidin-1-ium
2,2,2-trifluoroacetate
##STR00052##
[0352] Step 1: Synthesis of tert-butyl
2-(piperidin-1-yl)acetate
##STR00053##
[0354] To a stirred solution of piperidine (5.0 g, 58.71 mmol, 1.0
eq) in DCM (75 ml) at 0.degree. C. was added triethylamine (29.7 g,
293.55 mmol, 5.0 eq) followed by tert-butyl 2-chloroacetate (12.62
ml, 88.07 mmol, 1.5 eq). The reaction mixture was stirred at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was diluted with water (200 ml) and extracted with DCM (3.times.100
ml). The combined organic extracts were washed with water (100 ml),
brine solution (50 ml), dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to obtain the desired product
(10.0 g, yield: 85.47%) as a solid, which is used as such for next
step without further purification.
Step 2: Synthesis of 1-(carboxymethyl)piperidin-1-ium
2,2,2-trifluoroacetate
[0355] To a stirred solution of tert-butyl
2-(piperidin-1-yl)acetate (step 1, 10.0 g, 50.17 mmol, 1.0 eq) in
DCM (80 ml) at 0.degree. C. was added trifluoroacetic acid (20 ml).
The reaction mixture was allowed to stir at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was evaporated
under reduced pressure to obtain the desired product (12.9 g) as a
solid, which is used as such for next step without further
purification. ESI-MS: m/z 144.03 (M-TFA+H).sup.+.
Intermediate 11: Preparation of
2-(6-methylpyridin-3-yl)-1H-benzo[d]imidazole-5-carboxylic acid
##STR00054##
[0356] Step 1: Synthesis of methyl
2-(6-methylpyridin-3-yl)-1H-benzo[d]imidazole-5-carboxylate
##STR00055##
[0358] To a stirred solution of 6-methylnicotinic acid (1.23 g,
9.026 mmol, 1.5 eq) in DMF (10 ml) was added HBTU (3.47 g, 9.026
mmol, 1.5 eq), DIPEA (4.12 ml, 24.070 mmol, 4.0 eq) followed by
methyl 3,4-diaminobenzoate (1.0 g, 6.017 mmol, 1.0 eq). The
reaction mixture was stirred at room temperature for about 5 hours.
TLC indicated starting material was consumed and the desired
product was observed. The reaction mixture was diluted with water
(50 ml) and extracted with Ethyl acetate (2.times.100 ml). The
combined organic extracts were dried over Na.sub.2SO.sub.4,
filtered and evaporated under reduced pressure. The residue was
mixed with acetic acid (5 ml) and heated at 60.degree. C. for
overnight. Acetic acid was removed under reduced pressure and the
residue was treated with 10 ml of 5N sodium hydroxide solution. The
resulting solid was filtered and was washed with water and dried
under vacuum to obtain the desired product (0.804 g, yield: 50%) as
an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta.
ppm 13.4 (brs, 1H), 9.24 (d, J=1.5 Hz, 1H), 8.41 (dd, J=8.1, 2.1
Hz, 1H), 8.21 (brs, 1H), 7.87 (dd, J=8.4, 1.2 Hz, 1H), 7.71 (d,
J=8.4 Hz, 1H), 7.48 (d, J=8.1 Hz, 1H), 3.88 (s, 3H), 2.56 (s, 3H);
ESI-MS: m/z 267.93 (M+H).sup.+.
Step 2: Synthesis of
2-(6-methylpyridin-3-yl)-1H-benzo[d]imidazole-5-carboxylic acid
[0359] To a stirred solution of methyl
2-(6-methylpyridin-3-yl)-1H-benzo[d]imidazole-5-carboxylate (step
1, 0.8 g, 2.993 mmol, 1.0 eq) in MeOH (8 ml) and THF (8 ml) was
added aqueous 2.5N KOH solution (8.98 ml, 22.447 mmol, 7.5 eq). The
reaction mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure and diluted with water (15 ml), cooled to 0.degree. C., pH
adjusted to 6.0 with 1N HCl and extracted with 20% MeOH:DCM
(4.times.50 ml). The combined organic extracts were washed with
water (2.times.50 ml), dried over sodium sulfate, filtered and
evaporated under reduced pressure to obtain to obtain the desired
product (0.5 g, yield: 65.9%) as an off white solid. .sup.1H NMR
(300 MHz, DMSO-d.sub.6): .delta. ppm 13.38 (brs, 1H), 12.78 (brs,
1H), 9.24 (d, J=1.5 Hz, 1H), 8.41 (dd, J=8.1, 1.8 Hz, 1H), 8.25
(brs, 1H), 7.85 (m, 1H), 7.78-7.60 (m, 1H), 7.48 (d, J=8.1 Hz, 1H),
2.56 (s, 3H); ESI-MS: m/z 253.85 (M+H).sup.+.
Intermediate 12: Preparation of
2-(pyrazin-2-yl)-1H-benzo[d]imidazole-5-carboxylic acid
##STR00056##
[0360] Step 1: Synthesis of methyl
2-(pyrazin-2-yl)-1H-benzo[d]imidazole-5-carboxylate
##STR00057##
[0362] To a stirred solution of pyrazine-2-carboxylic acid (1.120
g, 9.026 mmol, 1.5 eq) in DMF (10 ml) was added HBTU (3.42 g, 9.026
mmol, 1.5 eq) followed by DIPEA (4.12 ml, 24.07 mmol, 4.0 eq). The
reaction mixture was stirred at room temperature for about 30
minutes, then methyl 3,4-diaminobenzoate (1.0 g, 6.017 mmol, 1.0
eq) was added and stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The reaction mixture was diluted with ice cooled
water (50 ml) and stirred at room temperature for about 15 minutes.
The precipitates formed were collected by filtration, washed with
water (50 ml) and dried under vacuum to obtain the solid. The
resulting solid was purified by silicagel column chromatography by
using 0-7% methanol in dichloromethane gradient. The fractions
containing the expected product were combined and concentrated
under reduced pressure to obtain the desired product (0.400 g,
yield: 26.3%) as an off-white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 9.52 (d, J=1.2 Hz, 1H), 8.78-8.69 (m,
2H), 8.25 (d, J=0.9 Hz, 1H), 7.79 (dd, J=8.7, 1.2 Hz, 1H), 7.67 (d,
J=8.4 Hz, 1H), 3.87 (s, 3H); ESI-MS: m/z 254.99 (M+H).sup.+.
Step 2: Synthesis of
2-(pyrazin-2-yl)-1H-benzo[d]imidazole-5-carboxylic acid
[0363] To a stirred solution of methyl
2-(pyrazin-2-yl)-1H-benzo[d]imidazole-5-carboxylate (step 1, 0.400
g, 1.573 mmol, 1.0 eq) in MeOH (4 ml) and THF (4 ml) was added
aqueous 2.5N KOH solution (4.71 ml, 11.79 mmol, 7.5 eq). The
reaction mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, diluted with water (5 ml), cooled to 0.degree. C., pH
adjusted to 6.0 with 1N HCl and extracted with 20% methanol in
dichloromethane (4.times.50 ml). The combined organic extracts were
washed with water (2.times.50 ml), dried over sodium sulfate,
filtered and evaporated under reduced pressure to obtain the
desired product (0.373 g, yield: 98%) as an off white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 13.68 (brs, 1H),
12.72 (brs, 1H), 9.53 (d, J=1.2 Hz, 1H), 8.88-8.80 (m, 2H),
8.33-8.20 (m, 1H), 7.79-7.60 (m, 2H).
Intermediate 13: Preparation of
4-(4-methyl-1H-imidazol-1-yl)benzoic acid
##STR00058##
[0364] Step 1: Synthesis of methyl
4-(4-methyl-1H-imidazol-1-yl)benzoate
##STR00059##
[0366] To a stirred solution of methyl 4-bromobenzoate (8.0 g,
37.20 mmol, 1.0 eq) in DMF (150 ml) was added CS.sub.2CO.sub.3
(24.24 g, 74.4 mmol, 2.0 eq) and 4-methyl-1H-imidazole (4.58 g,
55.8 mmol, 1.5 eq). The reaction mixture was bubbled through
nitrogen for about 20 minutes, then CuI (3.542 g, 18.6 mmol, 0.5
eq) was added and the mixture was again bubbled through nitrogen
for about 40 minutes. The reaction mixture was heated at
100.degree. C. for about 72 hours. TLC indicated starting material
was consumed and the desired product was observed. The reaction
mixture was allowed cool to room temperature and filtered through
celite. The filtrate was evaporated under reduced pressure and the
residue was purified by silicagel column chromatography by using
40% EtOAc in hexane eluent. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the desired product (2.413 g, yield: 30%) as an off-white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 8.14 (d,
J=8.7 Hz, 2H), 7.85 (s, 1H), 7.44 (d, J=8.7 Hz, 2H), 7.07 (s, 1H),
3.94 (s, 3H), 2.30 (s, 3H); ESI-MS: m/z 217.09 (M+H).sup.+.
Step 2: Synthesis of 4-(4-methyl-1H-imidazol-1-yl)benzoic acid
[0367] To a stirred solution of methyl
4-(4-methyl-1H-imidazol-1-yl)benzoate (step 1, 2.0 g, 9.248 mmol,
1.0 eq) in methanol (20 ml) was added 1N NaOH (75 ml, 73.99 mmol,
8.0 eq). The reaction mixture was stirred at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was cooled to
0.degree. C., pH adjusted to 7.0 with 1N HCl and evaporated under
reduced pressure to afford the desired product (1.5 g, yield:
80.2%) as a white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta.
ppm 9.50 (d, J=1.5 Hz, 1H), 8.27 (d, J=8.7 Hz, 2H), 7.90 (s, 1H),
7.84 (d, J=8.7 Hz, 2H), 2.47 (s, 3H); ESI-MS: m/z 203.12
(M+H).sup.+.
Intermediate 14: Preparation of 2-aminothiazole-4-carboxylic
acid
##STR00060##
[0369] To a stirred solution of ethyl 2-aminothiazole-4-carboxylate
(1.5 g, 8.71 mmol, 1.0 eq) in ethanol (15 ml) at 0.degree. C. was
added 1N NaOH solution (69.68 ml, 69.68 mmol, 8.0 eq). The reaction
mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The reaction mixture was evaporated under reduced
pressure to obtain the desired compound (3.0 g) as an off-white
solid.
Intermediate 15: Preparation of
4-(5-methyl-1,3,4-oxadiazol-2-yl)benzoic acid
##STR00061##
[0370] Step 1: Synthesis of methyl
4-(2-acetylhydrazine-1-carbonyl)benzoate
##STR00062##
[0372] To a stirred solution of 4-(methoxycarbonyl)benzoic acid
(5.0 g, 27.74 mmol, 1.0 eq) in DMF (50 ml) was added HBTU (15.7 g,
41.62 mmol, 1.5 eq), followed by triethyl amine (15.4 ml, 110.98
mmol, 4.0 eq). The reaction mixture was stirred at room temperature
for about 30 minutes then acetic hydrazide (3.0 g, 41.62 mmol, 1.5
eq) was added and stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The reaction mixture was diluted with water (500 ml)
and extracted with EtOAc (2.times.200 ml). The combined organic
extracts were dried over Na.sub.2SO.sub.4, filtered and evaporated
under reduced pressure to obtain the desired compound (5.0 g,
yield: 76%) as a colourless liquid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 10.5 (s, 1H), 9.97 (s, 1H), 8.07 (d,
J=8.1 Hz, 2H), 7.98 (d, J=8.4 Hz, 2H), 3.89 (s, 3H), 2.73 (s,
3H);
Step 2: Synthesis of methyl 4-(5-methyl-1, 3,
4-oxadiazol-2-yl)benzoate
##STR00063##
[0374] To a stirred solution of methyl
4-(2-acetylhydrazine-1-carbonyl)benzoate (step 1, 5.0 g, 21.186
mmol, 1.0 eq) in DCM (50 ml) at 0.degree. C. was added triethyl
amine (14.77 ml, 105.82 mmol, 5.0 eq) followed by
para-toluenesulphonyl chloride (6.0 g, 31.779 mmol, 1.5 eq). The
reaction mixture was allowed to stir at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was diluted with
DCM (200 ml) and washed with water (2.times.100 ml). The combined
organic extracts were dried over Na.sub.2SO.sub.4, filtered and
evaporated under reduced pressure. The crude compound (3.0 g) was
used as such for next step without further purification. .sup.1H
NMR (300 MHz, CDCl.sub.3): .delta. ppm 8.17 (d, J=8.7 Hz, 2H), 8.09
(d, J=8.7 Hz, 2H), 3.95 (s, 3H), 2.64 (s, 3H).
Step 3: Synthesis of 4-(5-methyl-1, 3, 4-oxadiazol-2-yl)benzoic
acid
[0375] To a stirred solution of methyl
4-(5-methyl-1,3,4-oxadiazol-2-yl)benzoate (step 2, 3.0 g, 13.761
mmol, 1.0 eq) in MeOH (30 ml) was added aqueous 1N NaOH solution
(110 ml, 110.097 mmol, 8.0 eq). The reaction mixture was stirred at
room temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was neutralized with 1N HCl and evaporated under reduced pressure.
To this compound acetonitrile (100 ml) was added and concentrated
under reduced pressure to obtain the crude desired product (3.0 g)
as a white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm
13.34 (brs, 1H), 8.12-8.10 (m, 4H), 2.61 (s, 3H); ESI-MS: m/z
226.87 (M+Na).sup.+.
Intermediate 16: Preparation of
4-(1,1-dioxidothiomorpholino)benzoic acid
##STR00064##
[0376] Step 1: Synthesis of Methyl
4-(1,1-dioxidothiomorpholino)benzoate
##STR00065##
[0378] To a stirred solution of methyl 4-bromobenzoate (1.5 g,
6.975 mmol, 1.0 eq) in toluene (30 ml) was added thiomorpholine
1,1-dioxide (1.13 g, 8.37 mmol, 1.2 eq) and Cs.sub.2CO.sub.3 (6.817
g, 20.92 mmol, 3.0 eq). The reaction mixture was purged with
nitrogen for about 10 minutes. Next, palladium acetate (0.015 g,
0.069 mmol, 0.01 eq) and BINAP (0.065 g, 0.104 mmol, 0.015 eq) were
added and the reaction mixture was degassed for about 30 minutes
and heated to 100.degree. C. for overnight. TLC indicated starting
material was consumed and the desired product was observed. The
reaction mixture was allowed to cool to room temperature, filtered
through celite pad and was washed with ethyl acetate (140 ml). The
organic layer was separated and was washed with saturated sodium
bicarbonate solution, dried over Na.sub.2SO.sub.4, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 90% ethyl acetate in
hexane as an eluent to obtain the desired product (1.2 g, yield:
63.8%) as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 7.82 (d, J=9.0 Hz, 2H), 7.09 (d, J=9.0 Hz, 2H), 3.92
(t, J=4.5 Hz, 4H), 3.78 (s, 3H), 3.14 (t, J=4.5 Hz, 4H); ESI-MS:
m/z 292.0 (M+Na).sup.+.
Step 2: Synthesis of 4-(1,1-dioxidothiomorpholino)benzoic acid
[0379] To a stirred solution of methyl
4-(1,1-dioxidothiomorpholino)benzoate (step 1, 1.6 g, 5.947 mmol,
1.0 eq) in MeOH (20 ml) was added aqueous 1N NaOH solution (47.5
ml, 47.5 mmol, 8.0 eq). The reaction mixture was stirred at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was neutralized with 1N HCl (47.5 ml) and evaporated under reduced
pressure. The crude solid was co-distilled with CH.sub.3CN (10 ml)
to obtain the desired product (2.0 g) as a white solid.
Intermediate 17: Preparation of
4-((1,1-dioxidothiomorpholino)methyl)benzoic acid
##STR00066##
[0380] Step 1: Synthesis of methyl
4-((1,1-dioxidothiomorpholino)methyl)benzoate
##STR00067##
[0382] To a stirred solution of methyl 4-(bromomethyl)benzoate (4.0
g, 17.46 mmol, 1.0 eq) in CH.sub.3CN (100 ml) was added
Cs.sub.2CO.sub.3 (17.0 g, 52.38 mmol, 3.0 eq). The reaction mixture
was stirred at room temperature for about 1 hour then
thiomorpholine 1,1-dioxide (2.36 g, 17.46 mmol, 1.0 eq) was added
and heated to reflux for overnight. TLC indicated starting material
was consumed and the desired product was observed. The reaction
mixture was cooled to room temperature, filtered through celite pad
and was washed with ethyl acetate (100 ml). The filtrate was washed
with water (100 ml), dried over Na.sub.2SO.sub.4, filtered and
evaporated under reduced pressure to obtain the desired product
(4.0 g) as a yellow oil. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 7.94 (d, J=8.1 Hz, 2H), 7.50 (d, J=8.4 Hz, 2H), 3.85
(s, 3H), 3.75 (s, 2H), 3.16-3.09 (m, 4H), 2.92-2.85 (m, 4H);
ESI-MS: m/z 306.03 (M+Na).sup.+.
Step 2: Synthesis of 4-((1,1-dioxidothiomorpholino)methyl)benzoic
acid
[0383] To a stirred solution of methyl
4-((1,1-dioxidothiomorpholino)methyl)benzoate (step 1, 1.0 g, 3.53
mmol, 1.0 eq) in MeOH (10 ml) and THF (10 ml) was added aqueous 1N
NaOH solution (28.2 ml, 28.2 mmol, 8.0 eq). The reaction mixture
was stirred at room temperature for overnight. TLC indicated
starting material was consumed and the desired product was
observed. The reaction mixture was neutralized with 1N HCl (28 ml)
and evaporated under reduced pressure to obtain the desired product
(1.5 g crude) as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 7.86 (d, J=8.1 Hz, 2H), 7.31 (d, J=7.8
Hz, 2H), 3.68 (s, 2H), 3.12-3.08 (m, 4H), 2.88-2.84 (m, 4H);
ESI-MS: m/z 268.06 (M-H).sup.-.
Intermediate 18: Preparation of
1-(4-chlorophenyl)cyclopropane-1-carbaldehyde
##STR00068##
[0384] Step 1: Synthesis of
(1-(4-chlorophenyl)cyclopropyl)methanol
##STR00069##
[0386] To a stirred solution of
1-(4-chlorophenyl)cyclopropane-1-carboxylic acid (Intermediate 8,
10 g, 51.02 mmol, 1.0 eq) in THF (150 ml) at 0.degree. C. under
nitrogen was added boranedimethyl sulphide (51 ml, 102.04 mmol, 2.0
eq, 2.0 M in THF). The reaction mixture was allowed to stir at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was cooled to 0.degree. C., quenched with saturated aqueous
ammonium chloride solution (50 ml) then diluted with water (100 ml)
and extracted with EtOAc (3.times.200 ml). The combined organic
extracts were washed with water (100 ml), brine solution (50 ml),
dried over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure to obtain the desired product (8.0 g) as colourless oil.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.36-7.27 (m, 4H),
3.65 (s, 2H), 0.86 (m, 4H).
Step 2: Synthesis of
1-(4-chlorophenyl)cyclopropane-1-carbaldehyde
[0387] To a stirred solution of
(1-(4-chlorophenyl)cyclopropyl)methanol (step 1, 6.0 g, 32.84 mmol,
1.0 eq) in DCM (60 ml) was added pyridinium chlorochromate (21.23
g, 98.54 mmol, 3.0 eq) and silicagel (21.23 g). The reaction
mixture was stirred at room temperature for about 1 hour. TLC
indicated starting material was consumed and the desired product
was observed. The reaction mixture was filtered through celite,
filtrate was diluted with water (150 ml) and extracted with DCM
(3.times.50 ml). The combined organic extracts were washed with
saturated sodium bicarbonate solution (2.times.30 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure to
obtain the desired product (6.0 g) as an oil, which is used as such
for next step without further purification.
Intermediate 19: Preparation of 5-isocyanato-2-methylpyridine
##STR00070##
[0389] To a stirred solution of 6-methylnicotinic acid (1.0 g,
7.299 mmol, 1.0 eq) in toluene (15 ml) was added triethylamine
(0.88 g, 8.759 mmol, 1.2 eq) and DPPA (2.4 g, 8.759 mmol, 1.2 eq).
The reaction mixture was stirred at room temperature for about 4
hours. TLC indicated starting material was consumed and the desired
product was observed. The reaction mixture was concentrated to
dryness. The residue was purified by silicagel column
chromatography by using 2% methanol in dichloromethane as an eluent
to obtain the desired product (0.75 g, yield: 77%) as an off-white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 9.09 (d,
J=1.8 Hz, 1H), 8.16 (dd, J=8.1, 2.1 Hz, 1H), 7.27 (d, J=8.1 Hz,
1H), 2.64 (s, 3H).
Intermediate 20: Preparation of 2-isocyanato-6-methylpyridine
##STR00071##
[0391] To a stirred solution of 6-methylpicolinic acid (1.5 g,
10.937 mmol, 1.0 eq) in toluene (20 ml) was added triethylamine
(1.328 g, 13.12 mmol, 1.2 eq) and diphenylphosphonic azide (3.61 g,
13.12 mmol, 1.2 eq). The reaction mixture was stirred at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was evaporated under reduced pressure to obtain the desired product
(0.6 g) as a oil, which is used as such for next step without
further purification.
Intermediate 21: Preparation of sodium
2-(dimethylamino)-1-hydroxyethane-1-sulfonate
##STR00072##
[0393] A stirred solution of 2,2-diethoxy-N,N-dimethylethan-1-amine
(15 g, 93.16 mmol. 1.0 eq), conc. hydrochloric acid (15.5 ml) and
water (7.5 ml) heated at 40.degree. C. for about 3 hours. Then a
solution of sodium pyro sulphite (15.93 g, 83.84 mmol, 0.9 eq)
dissolved in water (27 ml) was added dropwise and the mixture was
stirred for about 1 hour. Then 90 ml of ethanol was added to
reaction mixture and stirred for about 2 hours at 0.degree. C. The
suspension was filtered and washed with ethanol (20 ml), then dried
under vacuum at 40.degree. C. for about 30 minutes to afford the
desired product (15 g, yield: 84%) as a white solid.
Intermediate 22: Preparation of
4-(carboxymethyl)thiomorpholin-4-ium 1,1-dioxide
2,2,2-trifluoroacetate
##STR00073##
[0394] Step 1: Synthesis of tert-butyl
2-(1,1-dioxidothiomorpholino)acetate
##STR00074##
[0396] To a stirred solution of thiomorpholine 1,1-dioxide (3.0 g,
17.48 mmol) in DCM (30 ml) at 0.degree. C., were added TEA (17 ml,
122.37 mmol) and tert-butyl 2-chloroacetate (3.8 ml, 26.22 mmol).
The reaction mixture was stirred for overnight at room temperature.
After completion of the reaction (monitored by TLC), the reaction
mixture was diluted with DCM and washed with water, saturated
NH.sub.4Cl solution, brine and dried over Na.sub.2SO.sub.4. The
solvent was evaporated under reduced pressure and purified by
silica gel column (elution 1% MeOH in DCM) to afford the title
compound (3.0 g, yield: 68.96%) as a thick oil. H.sup.1 NMR
(DMSO-d.sub.6, 300 MHz): .delta. 3.35 (s, 2H), 3.06 (m, 8H), 1.41
(s, 9H).
Step 2: Synthesis of 4-(carboxymethyl)thiomorpholin-4-ium
1,1-dioxide 2,2,2-trifluoroacetate
[0397] To a stirred solution of tert-butyl
2-(1,1-dioxidothiomorpholino)acetate (step 1, 1.0 g, 4.01 mmol) in
DCM (10 ml), was added TFA (3 ml) and stirred for about 2 hours.
After completion of the reaction (monitored by TLC), the reaction
mixture was concentrated under reduced pressure to afford the
desired compound as a TFA salt (1.2 g, yield: 100%). Next reaction
was carried out without any further purification.
Examples
Example 1
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chlorobenzamido-
)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00075##
[0398] Step 1: Synthesis of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chlorobenzamido)-2-methylpro-
panamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7-
a,8, 9,10,11, ha,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl
(1S,3R)-2,2-dimethyl-3-(2-phenylacetoxy)cyclobutane-1-carboxylate
##STR00076##
[0400] To a stirred solution of 4-chlorobenzoic acid (0.304 g,
1.945 mmol, 1.5 eq) in DMF (10 ml) was added HBTU (0.737 g, 1.945
mmol, 1.5 eq) followed by DIPEA (1.55 ml, 9.07 mmol, 7.0 eq). The
reaction mixture was stirred at room temperature for about 30
minutes, then
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-iso
propyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,1-
1,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 1.0 g, 1.296 mmol, 1.0 eq) was added and stirred
at same temperature for overnight. TLC indicated starting material
was consumed and the desired product was observed. The reaction
mixture was diluted with water (150 ml) and extracted with ethyl
acetate (3.times.50 ml). The combined organic extracts were washed
with water (50 ml), brine solution (50 ml), dried over
Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure.
The residue was purified by silicagel column chromatography by
using 0-5% methanol in dichloromethane gradient. The fractions
containing the expected product were combined and concentrated
under reduced pressure to obtain the desired product (0.700 g,
yield: 59.3%) as a white solid. .sup.1H NMR (300 MHz, CDCl.sub.3):
.delta. ppm 7.71 (d, J=8.7 Hz, 2H), 7.41 (d, J=8.4 Hz, 2H),
7.37-7.31 (m, 5H), 7.09 (brs, 1H), 6.82 (brs, 1H), 5.14, 5.08 (ABq,
J.sub.AB=12.3 Hz, 2H), 4.43 (dd, J=11.4, 4.8 Hz, 1H), 3.21-3.10 (m,
1H), 2.85-2.72 (m, 3H), 2.70-2.57 (m, 2H), 2.37-2.23 (m, 2H),
2.08-1.72 (m, 6H), 1.68 (s, 3H), 1.66 (s, 3H), 1.62-1.37 (m, 6H),
1.34 (s, 3H), 1.32-1.28 (m, 2H), 1.28-1.16 (m, 8H), 1.16-1.06 (m,
1H), 1.03 (s, 3H), 0.95 (s, 3H), 0.93 (s, 3H), 0.86 (s, 3H), 0.84
(s, 3H), 0.83 (s, 3H), 0.82-0.78 (m, 1H); ESI-MS: m/z 931.5
(M+Na).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chloro
benzamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-o-
xo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
[0401] To a stirred solution of
(3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chlorobenz
amido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl
(1S,3R)-2,2-dimethyl-3-(2-phenylacetoxy)cyclobutane-1-carboxylate
(step 1, 0.700 g, 0.769 mmol, 1.0 eq) in MeOH (15 ml) and THF (15
ml) was added aqueous 2.5N KOH solution (2.15 ml, 5.383 mmol, 7.0
eq). The reaction mixture was stirred at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The organic phase was evaporated
under reduced pressure, the reaction mixture was diluted with water
(15 ml), cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (3.times.75 ml). The combined organic extracts
were washed with water (50 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-5% methanol in
dichloromethane gradient. The fractions containing the product were
combined and concentrated under reduced pressure to give a solid.
To this solid, acetonitrile (10 ml) was added and heated to reflux
for about 30 minutes. The mixture was cooled to 0.degree. C., solid
was filtered and was washed with n-hexane (10 ml) and dried under
vacuum to obtain the desired product (0.4 g, yield: 63.43%) as a
white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.72
(d, J=8.4 Hz, 2H), 7.42 (d, J=8.4 Hz, 2H), 7.09 (s, 1H), 6.77 (s,
1H), 4.46 (dd, J=11.1, 4.5 Hz, 1H), 3.22-3.11 (m, 1H), 2.86-2.76
(m, 3H), 2.71-2.53 (m, 2H), 2.38-2.25 (m, 2H), 2.10-2.0 (m, 1H),
1.98-1.73 (m, 5H), 1.69 (s, 3H), 1.68 (s, 3H), 1.65-1.40 (m, 6H),
1.37 (s, 3H), 1.34-1.30 (m, 2H), 1.28-1.18 (m, 8H), 1.16-0.99 (m,
1H), 1.06 (s, 3H), 1.04 (s, 3H), 0.94 (s, 3H), 0.88 (s, 3H), 0.86
(s, 3H), 0.85 (s, 3H), 0.84-0.78 (m, 1H); ESI-MS: m/z 841.5
(M+Na).sup.+; HPLC: 97.93%.
[0402] The below examples 2-21 were prepared by the procedure
similar (including reagents and reaction conditions) to the above
described in the synthesis of example-1 using with their
appropriate intermediates.
Example 2
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(1-(4-chlorophenyl-
)cyclopropane-1-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-o-
ctadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclob-
utane-1-carboxylic acid
##STR00077##
[0404] Intermediate 1 was coupled with
1-(4-chlorophenyl)cyclopropane-1-carboxylic acid (Intermediate 8)
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.59 (s, 1H), 7.37
(d, J=8.4 Hz, 2H), 7.31 (d, J=8.4 Hz, 2H), 5.46 (brs, 1H), 4.48
(dd, J=11.1, 4.5 Hz, 1H), 3.20-3.08 (m, 1H), 2.87-2.75 (m, 3H),
2.67-2.53 (m, 2H), 2.35-2.17 (m, 2H), 2.12-2.01 (m, 1H), 1.97-1.91
(m, 2H), 1.88-1.70 (m, 4H), 1.67-1.48 (m, 6H), 1.44-1.41 (m, 1H),
1.39 (s, 3H), 1.38 (s, 3H), 1.36 (s, 3H), 1.35-1.33 (m, 2H),
1.27-1.19 (m, 10H), 1.17 (s, 3H), 1.08 (s, 3H), 1.07-1.04 (m, 1H),
0.93 (s, 6H), 0.88 (s, 3H), 0.87 (s, 3H), 0.84-0.80 (m, 1H);
ESI-MS: m/z 881.3 (M+Na).sup.+; HPLC: 97.2%.
Example 3
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-benzamido-2-methyl-
propanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00078##
[0406] Intermediate 1 was coupled with benzoic acid followed by
hydrolysis gave the desired product as a white solid. .sup.1H NMR
(300 MHz, CDCl.sub.3): .delta. ppm 7.77 (d, J=7.2 Hz, 2H),
7.50-7.40 (m, 3H), 6.64 (brs, 1H), 4.46 (dd, J=11.1, 4.5 Hz, 1H),
3.22-3.11 (m, 1H), 2.88-2.53 (m, 5H), 2.39-2.24 (m, 2H), 2.10-2.0
(m, 1H), 2.0-1.85 (m, 3H), 1.81-1.72 (m, 2H), 1.69 (s, 3H), 1.67
(s, 3H), 1.65-1.42 (m, 7H), 1.37 (s, 3H), 1.36-1.30 (m, 2H),
1.27-1.21 (m, 7H), 1.20-1.15 (m, 1H), 1.07 (s, 3H), 1.02 (s, 3H),
0.94 (s, 3H), 0.86 (s, 6H), 0.84 (s, 3H), 0.82-0.78 (m, 1H);
ESI-MS: m/z 807.53 (M+Na).sup.+; HPLC: 95.3%.
Example 4
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(3,4-dichlorobenza-
mido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3-
a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta-
[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00079##
[0408] Intermediate 1 was coupled with 3,4-dichlorobenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.89 (d, J=2.1 Hz,
1H), 7.59 (dd, J=8.4, 1.8 Hz, 1H), 7.52 (d, J=8.4 Hz, 1H), 6.89
(brs, 1H), 6.85 (brs, 1H), 4.46 (dd, J=11.1, 4.5 Hz, 1H), 3.22-3.10
(m, 1H), 2.85-2.75 (m, 3H), 2.70-2.51 (m, 2H), 2.35-2.26 (m, 2H),
2.10-2.0 (m, 1H), 1.99-1.74 (m, 5H), 1.69 (s, 6H), 1.67-1.40 (m,
7H), 1.36 (s, 3H), 1.32 (m, 1H), 1.30-1.18 (m, 8H), 1.12-1.03 (m,
7H), 0.94 (s, 3H), 0.89 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H), 0.79
(m, 1H); ESI-MS: m/z 875.38 (M+Na).sup.+; HPLC: 98.06%.
Example 5
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(pyrazine-2-carboxamido)propanamido)-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00080##
[0410] Intermediate 1 was coupled with pyrazine-2-carboxylic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 9.39 (d, J=1.2 Hz,
1H), 8.80 (d, J=2.1 Hz, 1H), 8.56 (d, J=1.5 Hz, 1H), 8.19 (s, 1H),
7.46 (s, 1H), 4.45 (dd, J=11.1, 4.5 Hz, 1H), 3.21-3.10 (m, 1H),
2.87-2.53 (m, 5H), 2.38-2.25 (m, 2H), 2.11-1.80 (m, 5H), 1.80-1.62
(m, 3H), 1.70 (s, 3H), 1.68 (s, 3H), 1.60-1.42 (m, 5H), 1.41-1.28
(m, 3H), 1.37 (s, 3H), 1.28-1.12 (m, 8H), 1.07 (s, 3H), 0.94 (s,
3H), 0.93 (s, 3H), 0.90-0.75 (m, 10H); ESI-MS: m/z 809.39
(M+Na).sup.+; HPLC: 97.0%.
Example 6
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(6-aminonicotinami-
do)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00081##
[0412] Intermediate 1 was coupled with 6-aminonicotinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.15 (brs, 1H),
8.48 (d, J=1.8 Hz, 1H), 7.81 (s, 1H), 7.79 (d, J=1.8 Hz, 1H), 7.30
(s, 1H), 6.47 (brs, 2H), 6.40 (d, J=6.6 Hz, 1H), 4.38-4.32 (m, 1H),
3.13-3.03 (m, 1H), 2.85-2.73 (m, 3H), 2.43-2.26 (m, 4H), 2.07-2.03
(m, 1H), 1.92-1.80 (m, 3H), 1.78-1.64 (m, 2H), 1.62-1.43 (m, 4H),
1.39 (s, 3H), 1.38 (s, 3H), 1.35-1.17 (m, 4H), 1.26 (s, 3H),
1.15-1.10 (m, 6H), 1.07-0.94 (m, 3H), 0.91 (s, 3H), 0.88 (s, 3H),
0.85 (s, 3H), 0.82 (s, 3H), 0.81 (s, 3H), 0.80 (s, 3H), 0.84-0.77
(m, 1H); ESI-MS: m/z 823.49 (M+Na).sup.+; HPLC: 97.38%.
Example 7
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(5-methylpyrazine-2-carboxamido)propanamido)--
2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-c-
yclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxyli-
c acid
##STR00082##
[0414] Intermediate 1 was coupled with
5-methylpyrazine-2-carboxylic acid followed by hydrolysis gave the
desired product as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 9.25 (d, J=0.9 Hz, 1H), 8.402 (d, J=0.6
Hz, 1H), 8.10 (s, 1H), 7.54 (s, 1H), 4.46 (dd, J=8.4, 3.3 Hz, 1H),
3.20-3.11 (m, 1H), 2.85-2.77 (m, 3H), 2.70-2.55 (m, 2H), 2.69 (s,
3H), 2.36-2.25 (m, 2H), 2.08-1.82 (m, 4H), 1.80-1.70 (m, 2H), 1.69
(s, 3H), 1.66 (s, 3H), 1.65-1.57 (m, 2H), 1.55-1.40 (m, 4H), 1.37
(s, 3H), 1.36-1.25 (m, 3H), 1.25-1.17 (m, 7H), 1.17-1.02 (m, 1H),
1.07 (s, 3H), 0.97 (s, 3H), 0.92 (s, 3H), 0.90-0.82 (m, 9H),
0.81-0.78 (m, 1H); ESI-MS: m/z 823.49 (M+Na).sup.+; HPLC:
96.29%.
Example 8
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-1-(tert-butox-
ycarbonyl)pyrrolidine-2-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,
5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,1-
3,13a-octadeca
hydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane--
1-carboxylic acid
##STR00083##
[0416] Intermediate 1 was coupled with
(tert-butoxycarbonyl)-L-proline followed by hydrolysis gave the
desired product as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 12.13 (brs, 1H), 8.18 (s, 1H), 6.75 (s,
1H), 4.39-4.33 (m, 1H), 4.05-3.99 (m, 1H), 3.36 (m, 1H), 3.26 (m,
1H), 3.13-3.06 (m, 1H), 2.84-2.64 (m, 3H), 2.42-2.15 (m, 4H),
2.10-1.83 (m, 5H), 1.80-1.63 (m, 5H), 1.62-1.48 (m, 4H), 1.45-1.23
(m, 22H), 1.22-1.02 (m, 13H), 0.93-0.80 (m, 15H); ESI-MS: m/z
900.53 (M+Na).sup.+; HPLC: 98.3%.
Example 9
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(4-ethylpiperaz-
in-1-yl)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00084##
[0418] Intermediate 1 was coupled with 2,2,2-trifluoroacetate salt
of 2-(4-ethylpiperazin-1-yl)acetic acid (Intermediate 9) followed
by hydrolysis gave the desired product as a white solid. .sup.1H
NMR (300 MHz, CDCl.sub.3+CD.sub.3OD): .delta. ppm 7.17 (s, 1H),
4.50-4.42 (m, 1H), 3.26-3.22 (m, 2H), 3.20-3.08 (m, 10H), 3.0-2.90
(m, 2H), 2.86-2.75 (m, 2H), 2.66-2.53 (m, 2H), 2.36-2.20 (m, 2H),
2.10-1.86 (m, 3H), 1.85-1.66 (m, 2H), 1.57 (s, 3H), 1.54 (s, 3H),
1.50-1.34 (m, 16H), 1.28-1.18 (m, 11H), 1.13 (s, 3H), 1.05 (s, 3H),
1.0-0.80 (m, 10H); ESI-MS: m/z 835.6 (M+H).sup.+; HPLC: 95.03%.
Example 10
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(2-(piperidin-1-yl)acetamido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00085##
[0420] Intermediate 1 was coupled with
1-(carboxymethyl)piperidin-1-ium 2,2,2-trifluoroacetate
(Intermediate 10) followed by hydrolysis gave the desired product
as a white solid. .sup.1H NMR (300 MHz, pyridine-d5): .delta. ppm
8.33 (s, 1H), 8.30 (s, 1H), 4.73 (dd, J=11.1, 4.5 Hz, 1H),
3.25-2.98 (m, 7H), 2.73-2.56 (m, 2H), 2.40-2.35 (m, 4H), 2.25-2.08
(m, 2H), 1.93-1.75 (m, 2H), 1.85 (s, 3H), 1.78 (s, 3H), 1.75-1.62
(m, 3H), 1.57 (s, 3H), 1.53-1.36 (m, 17H), 1.35-1.22 (m, 9H),
1.21-1.05 (m, 2H), 1.05-0.88 (m, 13H), 0.84-0.76 (m, 1H); ESI-MS:
m/z 806.5 (M+H).sup.+; HPLC: 91.5%.
Example 11
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-amino-2-methylp-
ropanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-o-
xo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cycl-
openta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid hydrochloride
##STR00086##
[0421] Step 1: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-((tert-butoxyca-
rbonyl)amino)-2-methylpropanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,-
8,
8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,1-
3a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcy-
clobutane-1-carboxylic acid
##STR00087##
[0423] Intermediate 1 was coupled with
2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (Intermediate
1-step 9) followed by hydrolysis gave the desired product as an
off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm
7.18 (s, 1H), 6.39 (s, 1H), 4.87 (s, 1H), 4.47 (dd, J=11.1, 4.5 Hz,
1H), 3.22-3.12 (m, 1H), 2.96-2.74 (m, 3H), 2.67-2.48 (m, 3H), 2.20
(d, J=18.6 Hz, 1H), 2.11-1.90 (m, 4H), 1.83-1.51 (m, 9H), 1.50-1.28
(m, 23H), 1.28-1.20 (m, 8H), 1.15 (s, 3H), 1.07 (s, 3H), 0.99-0.83
(m, 16H).
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-amino-2-methylp-
ropanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-o-
xo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cycl-
openta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid hydrochloride
[0424] To a stirred solution of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-((tert-butoxy
carbonyl)amino)-2-methylpropanamido)-2-methylpropanamido)-1-isopropyl-5a,-
5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13-
,13a-octadeca
hydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane--
1-carboxylic acid (step 1, 0.600 g, 0.692 mmol, 1.0 eq) in a round
bottomed flask at 0.degree. C. was added 3N HCl in 1,4-dioxane (5
ml). The reaction mixture was allowed to stir at room temperature
for overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was evaporated
under reduced pressure, the residue was washed with n-hexane (10
ml) and dried under vacuum to obtain the solid. To this solid
compound, MTBE (10 ml) was added and heated to reflux for 30
minutes. The mixture was cooled to 0.degree. C., filtered, solid
was washed with MTBE and dried under vacuum to obtain the desired
product (0.360 g, yield: 64.75%) as a white solid. .sup.1H NMR (300
MHz, CD.sub.3OD): .delta. ppm 7.19 (s, 1H), 4.48-4.41 (m, 1H), 3.21
(m, 1H), 2.95-2.80 (m, 3H), 2.63 (d, J=18.6 Hz, 1H), 2.57-2.42 (m,
1H), 2.40-2.28 (m, 1H), 2.23-2.14 (m, 1H), 2.10-1.77 (m, 4H),
1.76-1.60 (m, 4H), 1.60-1.50 (m, 15H), 1.50-1.38 (m, 2H), 1.35 (s,
3H), 1.32-1.28 (m, 3H), 1.27-1.18 (m, 9H), 1.18-1.05 (m, 1H),
1.05-0.97 (m, 9H), 0.95-0.87 (m, 7H); ESI-MS: m/z 788.51
(M-HCl+Na).sup.+; HPLC: 93.5%, Cl.sup.- ion content by Ion
chromatography: 5.2%
Example 12
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(1H-benzo[d]imidaz-
ole-5-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00088##
[0426] Intermediate 1 was coupled with
1H-benzo[d]imidazole-5-carboxylic acid followed by hydrolysis gave
the desired product as a white solid. .sup.1H NMR (300 MHz,
CD.sub.3OD): .delta. ppm 8.29 (d, J=8.4 Hz, 1H), 8.22 (brs, 1H),
7.84 (dd, J=8.4, 1.2 Hz, 1H), 7.67 (d, J=8.4 Hz, 1H), 7.50 (brs,
1H), 4.47-4.38 (m, 1H), 3.28-3.18 (m, 1H), 3.0-2.92 (m, 1H),
2.88-2.77 (m, 2H), 2.67 (d, J=18.6 Hz, 1H), 2.53-2.42 (m, 1H),
2.38-2.28 (m, 1H), 2.21 (d, J=18.6 Hz, 1H), 2.02-1.88 (m, 4H),
1.82-1.72 (m, 1H), 1.70-1.62 (m, 2H), 1.58 (brs, 6H), 1.56-1.40 (m,
4H), 1.38-1.27 (m, 8H), 1.27-1.10 (m, 7H), 1.01 (s, 3H), 0.96 (s,
3H), 0.92 (s, 3H), 0.90 (s, 3H), 0.90-0.78 (m, 7H); ESI-MS: m/z
823.62 (M-H).sup.-; HPLC: 96.3%.
Example 13
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(2-(6-methylpyridin-3-yl)-
-1H-benzo[d]imidazole-5-carboxamido)propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,-
7a,8,9,10,11,11a,11b,12,
13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimet-
hylcyclobutane-1-carboxylic acid
##STR00089##
[0428] Intermediate 1 was coupled with
2-(6-methylpyridin-3-yl)-1H-benzo[d]imidazole-5-carboxylic acid
(Intermediate 11) followed by hydrolysis gave the desired product
as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta.
ppm 13.2 (brs, 1H), 12.1 (brs, 1H), 9.23 (d, J=1.8 Hz, 1H), 8.40
(dd, J=8.1, 2.1 Hz, 1H), 8.14 (brs, 1H), 7.78 (d, J=7.8 Hz, 1H),
7.75-7.55 (m, 1H), 7.47 (d, J=8.1 Hz, 1H), 7.34 (brs, 1H),
4.38-4.30 (m, 1H), 3.15-3.05 (m, 1H), 2.88-2.70 (m, 4H), 2.56 (s,
3H), 2.35-2.22 (m, 3H), 2.14-2.05 (m, 1H), 1.95-1.80 (m, 4H),
1.80-1.60 (m, 2H), 1.60-1.50 (m, 2H), 1.48 (s, 3H), 1.47 (s, 3H),
1.42-1.20 (m, 5H), 1.32 (s, 3H), 1.20-1.10 (m, 8H), 1.08-0.93 (m,
2H), 0.90 (s, 3H), 0.87 (s, 3H), 0.86 (s, 3H), 0.79 (s, 3H), 0.77
(s, 3H), 0.76 (s, 3H); ESI-MS: m/z 939.03 (M+Na).sup.+; HPLC:
97.12%.
Example 14
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2,4-dimethylthiaz-
ole-5-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00090##
[0430] Intermediate 1 was coupled with
2,4-dimethylthiazole-5-carboxylic acid followed by hydrolysis gave
the desired product as an off-white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 7.01 (s, 1H), 6.26 (s, 1H), 4.46 (dd,
J=11.4, 4.8 Hz, 1H), 3.19-3.10 (m, 1H), 2.88-2.75 (m, 3H), 2.67 (s,
3H), 2.64 (s, 3H), 2.63-2.56 (m, 2H), 2.40-2.26 (m, 2H), 2.12-1.82
(m, 5H), 1.82-1.50 (m, 8H), 1.65 (s, 3H), 1.64 (s, 3H), 1.49-1.30
(m, 2H), 1.37 (s, 3H), 1.28-1.18 (m, 7H), 1.07 (m, 7H), 0.94 (s,
3H), 0.89 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H), 0.99-0.78 (m, 1H);
ESI-MS: m/z 842.51 (M+Na).sup.+; HPLC: 95.6%.
Example 15
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(2-(pyrazin-2-yl)-1H-benz-
o[d]imidazole-5-carboxamido)propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,1-
0,11,11a,11b,12,13,13a-octa
decahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobut-
ane-1-carboxylic acid
##STR00091##
[0432] Intermediate 1 was coupled with
2-(pyrazin-2-yl)-1H-benzo[d]imidazole-5-carboxylic acid
(Intermediate 12) followed by hydrolysis gave the desired product
as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta.
ppm 13.65 (s, 0.5H), 13.55 (s, 0.5H), 12.1 (s, 1H), 9.52 (s, 1H),
8.85-8.79 (m, 2H), 8.42 (s, 0.5H), 8.22-8.17 (m, 1H), 8.09 (s,
0.5H), 7.86-7.76 (m, 1H), 7.58 (d, J=8.4 Hz, 1H), 4.38-4.30 (m,
1H), 3.14-3.06 (m, 1H), 2.87-2.72 (m, 3H), 2.35-2.22 (m, 3H),
2.14-2.05 (m, 2H), 1.95-1.72 (m, 4H), 1.70-1.55 (m, 3H), 1.48 (s,
3H), 1.46 (s, 3H), 1.44-1.30 (m, 5H), 1.28-1.20 (m, 3H), 1.26 (s,
3H), 1.16-1.10 (m, 6H), 1.05-0.95 (m, 1H), 0.90 (s, 3H), 0.86 (m,
6H), 0.82-0.75 (m, 10H); ESI-MS: m/z 903.5 (M+H).sup.+; HPLC:
96.6%.
Example 16
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(1-methyl-1H-imidazole-2--
carboxamido)
propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobuta-
ne-1-carboxylic acid
##STR00092##
[0434] Intermediate 1 was coupled with
1-methyl-1H-imidazole-2-carboxylic acid followed by hydrolysis gave
the desired product as an off-white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 7.61 (m, 1H), 7.04 (s, 1H), 7.0 (d, J=6.6
Hz, 1H), 4.46 (dd, J=11.1, 4.5 Hz, 1H), 4.02 (s, 3H), 3.20-3.08 (m,
1H), 2.88-2.52 (m, 5H), 2.43-2.22 (m, 2H), 2.10-2.0 (m, 1H),
1.98-1.82 (m, 4H), 1.82-1.40 (m, 14H), 1.37 (s, 3H), 1.36-1.18 (m,
9H), 1.07 (s, 3H), 1.05-1.0 (m, 1H), 0.95 (s, 3H), 0.92 (s, 3H),
0.89-0.83 (m, 9H), 0.78 (m, 1H); ESI-MS: m/z 789.7 (M+H).sup.+;
HPLC: 93.5%.
Example 17
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-3a-(2-(3-isopropyl-1H-pyrazole-5-carboxamido)-2-methylpropanamido)-
-5a,5b,8,8,
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-o-
ctadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclob-
utane-1-carboxylic acid
##STR00093##
[0436] Intermediate 1 was coupled with
3-isopropyl-1H-pyrazole-5-carboxylic acid followed by hydrolysis
gave the desired product as an off-white solid. .sup.1H NMR (300
MHz, CDCl.sub.3): .delta. ppm 7.79 (s, 1H), 7.14 (s, 1H), 6.58 (s,
1H), 4.46 (dd, J=11.1, 4.5 Hz, 1H), 3.18-3.10 (m, 1H), 3.10-2.90
(m, 1H), 2.90-2.75 (m, 3H), 2.75-2.52 (m, 2H), 2.40-2.20 (m, 2H),
2.10-2.0 (m, 1H), 2.0-1.85 (m, 4H), 1.78-1.69 (m, 2H), 1.65 (s,
3H), 1.60 (s, 3H), 1.58-1.40 (m, 6H), 1.37 (s, 3H), 1.32 (s, 3H),
1.29 (s, 3H), 1.27-1.20 (m, 9H), 1.17-1.10 (m, 1H), 1.07 (s, 3H),
0.99 (s, 3H), 0.92 (s, 3H), 0.88-0.77 (m, 10H); ESI-MS: m/z 817.7
(M+H).sup.+; HPLC: 96.5%.
Example 18
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(4-morpholinobenzamido)pr-
opanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadec-
ahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane--
1-carboxylic acid
##STR00094##
[0438] Intermediate 1 was coupled with 4-morpholinobenzoic acid
followed by hydrolysis gave the desired product as an off-white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.69 (d,
J=8.4 Hz, 2H), 6.89 (d, J=9.0 Hz, 2H), 6.43 (s, 1H), 4.46 (dd,
J=11.1, 4.5 Hz, 1H), 3.90-3.83 (m, 4H), 3.30-3.23 (m, 4H),
3.20-3.09 (m, 1H), 2.90-2.75 (m, 3H), 2.70-2.52 (m, 2H), 2.38-2.23
(m, 2H), 2.10-2.0 (m, 1H), 2.0-1.82 (m, 3H), 1.82-1.70 (m, 2H),
1.67 (s, 3H), 1.63 (s, 3H), 1.61-1.54 (m, 2H), 1.54-1.40 (m, 3H),
1.39-1.29 (m, 3H), 1.37 (s, 3H), 1.28-1.17 (m, 9H), 1.06 (s, 3H),
1.02 (s, 3H), 0.93 (s, 3H), 0.89-0.82 (m, 9H), 0.78 (m, 1H);
ESI-MS: m/z 892.76 (M+Na).sup.+; HPLC: 98.1%.
Example 19
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(3,5-dimethylisoxa-
zole-4-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00095##
[0440] Intermediate 1 was coupled with
3,5-dimethylisoxazole-4-carboxylic acid followed by hydrolysis gave
the desired product as an off-white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 6.75 (s, 1H), 6.48 (s, 1H), 4.46 (dd,
J=11.1, 4.5 Hz, 1H), 3.20-3.10 (m, 1H), 2.89-2.51 (m, 5H), 2.60 (s,
3H), 2.42 (s, 3H), 2.40-2.27 (m, 2H), 2.10-2.0 (m, 1H), 1.98-1.80
(m, 3H), 1.78-1.72 (m, 2H), 1.69 (s, 3H), 1.66 (s, 3H), 1.63-1.51
(m, 3H), 1.50-1.39 (m, 3H), 1.36 (s, 3H), 1.35-1.31 (m, 2H),
1.29-1.18 (m, 8H), 1.12 (s, 3H), 1.05 (s, 3H), 1.01 (m, 1H), 0.95
(s, 3H), 0.90 (s, 3H), 0.87 (s, 3H), 0.85 (s, 3H), 0.80 (m, 1H);
ESI-MS: m/z 804.52 (M+H).sup.+; HPLC: 97.6%.
Example 20
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(4-(4-methyl-1H-imidazol--
1-yl)benz
amido)propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b-
,12,13,13a-octadeca
hydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane--
1-carboxylic acid
##STR00096##
[0442] Intermediate 1 was coupled with
4-(4-methyl-1H-imidazol-1-yl)benzoic acid (Intermediate 13)
followed by hydrolysis gave the desired product as an off-white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.89 (d,
J=8.4 Hz, 2H), 7.43 (d, J=8.4 Hz, 2H), 7.08-6.97 (m, 2H), 4.46 (dd,
J=11.1, 4.5 Hz, 1H), 3.22-3.11 (m, 1H), 2.90-2.53 (m, 5H),
2.35-2.27 (m, 2H), 2.30 (s, 3H), 2.10-1.84 (m, 4H), 1.78-1.68 (m,
7H), 1.67-1.18 (m, 17H), 1.37 (s, 3H), 1.12-1.05 (m, 7H), 0.95 (s,
3H), 0.87 (s, 3H), 0.85 (s, 3H), 0.84 (s, 3H), 0.79 (m, 1H);
ESI-MS: m/z 865.72 (M+H).sup.+; HPLC: 96.8%.
Example 21
Preparation of
(1S,3R)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-chlorobenzamido-
)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00097##
[0444] Intermediate 2 was coupled with 4-chlorobenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.72 (d, J=8.1 Hz,
2H), 7.42 (d, J=7.8 Hz, 2H), 7.07 (s, 1H), 6.78 (s, 1H), 4.47 (dd,
J=11.4, 4.5 Hz, 1H), 3.23-3.10 (m, 1H), 2.86-2.76 (m, 3H),
2.72-2.50 (m, 2H), 2.40-2.25 (m, 2H), 2.13-2.04 (m, 1H), 2.04-1.82
(m, 4H), 1.80-1.66 (m, 7H), 1.64-1.45 (m, 6H), 1.44-1.41 (m, 1H),
1.40-1.31 (m, 2H), 1.36 (s, 3H), 1.30-1.20 (m, 7H), 1.20-1.09 (m,
1H), 1.04 (s, 6H), 0.95 (s, 3H), 0.88 (s, 3H), 0.857 (s, 3H), 0.851
(s, 3H), 0.79 (m, 1H); ESI-MS: m/z 841.52 (M+Na).sup.+; HPLC:
94.19%.
Example 22
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-fluorobenzamido-
)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00098##
[0445] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-fluorobenz
amido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl) (1R,3S)-2, 2-dimethylcyclobutane-1,3-dicarboxylate
##STR00099##
[0447] To a stirred solution of 4-fluorobenzoic acid (0.54 g, 3.89
mmol, 1.5 eq) in DMF (20 ml) at 0.degree. C. under nitrogen
atmosphere was added EDCI (0.99 g, 5.18 mmol, 2 eq), HOBT (0.52 g,
3.89 mmol, 1.5 eq), DMAP (0.15 g, 1.29 mmol, 0.5 eq) and added
Triethylamine (1.08 ml, 7.78 mmol, 3 eq) and stirred it for about
30 minutes. Then added 1-((3aR,5aR,5bR,
7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-isopropyl-5a,5b-
,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,1-
3a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl) 3-benzyl
(1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate (Intermediate 1,
2.0 g, 2.59 mmol, 1 eq). The reaction mixture was stirred at room
temperature for about 14 hours. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was quenched with ice cold water, filtered through Buchner funnel.
The solid was separated, was dissolved in DCM, washed with sodium
bicarbonate, water, brine solution then dried over sodium sulfate
and concentrated under reduced pressure to give a crude compound.
The crude compound was purified by flash silica column
chromatography (100-200 silica gel) using 2% MeOH in DCM as an
eluent gave the desired product (1.5 g, yield: 65.21%) as an
off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm
8.16 (s, 1H), 7.94-7.98 (m, 2H), 7.35 (m, 5H), 7.27-7.34 (m, 3H),
5.15, 5.10 (ABq, J.sub.AB=12.3 Hz, 2H), 4.32-4.36 (t, 1H),
3.07-3.10 (t, 1H), 2.75-2.88 (m, 4H), 2.39-2.43 (m, 1H), 2.27-2.30
(m, 2H), 2.04-2.09 (m, 1H), 1.84-1.90 (m, 3H), 1.48-1.75 (m, 6H),
1.41-1.42 (m, 6H), 1.23-1.32 (m, 9H), 1.11-1.13 (m, 5H), 0.90-1.05
(m, 3H), 0.87 (s, 3H), 0.85 (s, 3H), 0.83 (s, 3H), 0.79-0.80 (m,
9H); ESI-MS: m/z 915.54 (M+Na).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-fluorobenz
amido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,
6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen--
9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
[0448] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-fluorobenzamido)-2-methyl-
propanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(step 1, 1.5 g, 0.541 mmol, 1.0 eq) in MeOH (15 ml) and THF (15 ml)
was added aqueous 2.5 N KOH solution (3.35 ml, 8.39 mmol, 5 eq).
The reaction mixture was stirred at room temperature for overnight.
TLC indicated starting material was consumed and the desired
product was observed. The organic phase was evaporated under
reduced pressure, the reaction mixture was diluted with water (10
ml), cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (2.times.75 ml). The combined organic extracts
were washed with water (50 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-5% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the solid. The solid compound was dissolved in MTBE (10 ml)
and added Hexane (30 ml) then precipitate was formed, and filtered
through Buchner funnel. The solid was washed with MTBE:Hexane (1:3,
10 ml) and dried under vacuum to obtain the desired product (200
mg, yield: 15.38%) as a white solid. .sup.1H NMR (400 MHz,
DMSO-d6): .delta. ppm 12.14 (s, 1H), 8.16 (s, 1H), 7.94-7.98 (m,
2H), 7.27-7.34 (m, 3H), 4.32-4.36 (t, 1H), 3.07-3.10 (t, 1H),
2.75-2.88 (m, 4H), 2.39-2.43 (m, 1H), 2.27-2.30 (m, 2H), 2.04-2.09
(m, 1H), 1.84-1.90 (m, 3H), 1.48-1.75 (m, 6H), 1.41-1.42 (m, 6H),
1.23-1.32 (m, 9H), 1.11-1.13 (m, 5H), 0.90-1.05 (m, 3H), 0.87 (s,
3H), 0.85 (s, 3H), 0.83 (s, 3H), 0.79-0.80 (m, 9H); ESI-MS: m/z
803.47 (M+H).sup.+; HPLC: 92.89%.
[0449] The below examples 23-50 were prepared by the procedure
similar (including reagents and reaction conditions) to the above
described in the synthesis of example-22 using with their
appropriate intermediates.
Example 23
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(4-methylbenzamido)propan-
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahyd-
ro-2H-cyclopenta[a]chrysen-9-yl)
oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
##STR00100##
[0451] Intermediate 1 was coupled with 4-methylbenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (400 MHz, DMSO-d6): .delta. ppm 12.13 (s, 1H), 8.03 (s,
1H), 7.78 (d, J=8.0 Hz, 2H), 7.31 (s, 1H), 7.25 (d, J=8.0 Hz, 2H),
4.32-4.36 (m, 1H), 3.07-3.10 (m, 1H), 2.73-2.81 (m, 3H), 2.39-2.44
(m, 1H), 2.35 (s, 3H), 2.25-2.30 (m, 2H), 2.04-2.08 (m, 1H),
1.86-1.95 (m, 3H), 1.51-1.75 (m, 6H), 1.425 (d, J=8.0 Hz, 6H),
1.23-1.29 (m, 8H), 1.11-1.13 (m, 7H), 0.95-1.02 (m, 3H), 0.91 (s,
3H), 0.865 (d, J=8.0 Hz, 6H), 0.80-0.81 (m, 9H); ESI-MS: m/z 799.42
(M+H).sup.+; HPLC: 91.77%.
Example 24
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(furan-3-carboxami-
do)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,-
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a-
]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00101##
[0453] Intermediate 1 was coupled with furan-3-carboxylic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.15 (s, 1H),
8.215 (d, J=3 Hz, 1H), 7.83 (s, 1H), 7.715 (d, J=3 Hz, 1H), 7.35
(s, 1H), 6.835 (d, J=3 Hz, 1H), 4.33-4.38 (m, 1H), 3.07-3.11 (m,
1H), 2.73-2.83 (m, 3H), 2.28-2.43 (m, 3H), 2.03-2.10 (m, 1H),
1.86-1.90 (m, 3H), 1.40-1.76 (m, 6H), 1.34-1.38 (m, 6H), 1.23-1.30
(m, 8H), 1.11-1.19 (m, 9H), 1.01 (m, 2H), 0.95 (s, 3H), 0.87-0.91
(s, 3H), 0.81-0.85 (m, 11H); ESI-MS: m/z 797.42 (M+Na).sup.+; HPLC:
92.55%.
Example 25
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(4-(trifluoromethyl)benza-
mido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00102##
[0455] Intermediate 1 was coupled with 4-(trifluoromethyl)benzoic
acid followed by hydrolysis gave the desired product as an off
white solid. .sup.1H NMR (300 MHz, DMSO): .delta. ppm 12.13 (s,
1H), 8.39 (s, 1H), 8.10 (d, J 8.4 Hz, 2H), 7.85 (d, J 8.4 Hz, 2H),
7.39 (s, 1H), 4.31-4.36 (t, 1H), 3.07-3.11 (t, 1H), 2.72-2.82 (m,
3H), 2.27-2.31 (m, 3H), 2.05-2.11 (m, 2H), 1.76-1.89 (m, 5H),
1.56-1.63 (m, 4H), 1.43-1.44 (m, 9H), 1.26-1.30 (m, 8H), 1.11-1.13
(m, 6H), 0.98-1.03 (m, 3H), 0.85-0.90 (m, 6H), 0.76-0.80 (m, 9H);
ESI-MS: m/z 853.43 (M+H).sup.+. HPLC: 97.60%.
Example 26
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(furan-2-carboxami-
do)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,-
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a-
]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00103##
[0457] Intermediate 1 was coupled with furan-2-carboxylic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.13 (s, 1H),
7.83-7.91 (m, 2H), 7.45 (s, 1H), 7.08-7.10 (m, 1H), 6.61-6.63 (m,
1H), 4.33-4.38 (m, 1H), 3.07-3.11 (m, 1H), 2.73-2.83 (m, 3H),
2.28-2.43 (m, 3H), 2.03-2.10 (m, 1H), 1.86-1.90 (m, 3H), 1.40-1.76
(m, 6H), 1.34-1.38 (m, 6H), 1.23-1.30 (m, 8H), 1.11-1.19 (m, 9H),
1.01 (m, 2H), 0.95 (s, 3H), 0.87-0.91 (s, 3H), 0.81-0.85 (m, 11H);
ESI-MS: m/z 797.42 (M+Na).sup.+; HPLC: 92.65%.
Example 27
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(1-phenylcyclopentane-1-c-
arboxamido)
propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00104##
[0459] Intermediate 1 was coupled with
1-phenylcyclopentane-1-carboxylic acid followed by hydrolysis gave
the desired product as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 12.03 (brs, 1H), 7.20-7.36 (m, 6H), 6.48
(s, 1H), 4.33-4.38 (m, 1H), 3.07-3.09 (m, 1H), 2.70-2.82 (m, 3H),
2.28-2.31 (m, 2H), 1.50-2.03 (m, 16H), 1.27-1.36 (m, 15H),
1.03-1.10 (m, 14H), 0.81-0.96 (m, 16H); ESI-MS: m/z 853.56
(M+H).sup.+; HPLC: 98.50%.
Example 28
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(quinoline-2-carboxamido)-
propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octad-
ecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutan-
e-1-carboxylic acid
##STR00105##
[0461] Intermediate 1 was coupled with quinoline-2-carboxylic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.13 (s, 1H),
8.98 (s, 1H), 8.59 (d, J=8.7 Hz, 1H), 8.16 (d, J=8.7 Hz, 1H),
8.119-8.092 (m, 2H), 7.90-7.85 (m, 1H), 7.75-7.70 (m, 2H),
4.37-4.32 (t, 1H), 3.12-3.07 (m, 2H), 2.85-2.73 (m, 3H), 2.40-2.12
(m, 5H), 1.92-1.79 (m, 5H), 1.60 (m, 9H), 1.31-1.26 (m, 6H),
1.17-1.10 (m, 8H), 0.91-0.88 (m, 11H), 0.81-0.79 (m, 10H); ESI-MS:
m/z 858.55 (M+Na).sup.+. HPLC: 97.06%.
Example 29
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(3-methylpicolinamido)pro-
panamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadeca-
hydro-2H-cyclopenta[a]chrysen-9-yl)
oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
##STR00106##
[0463] Intermediate 1 was coupled with 3-methylpicolinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.18 (brs, 1H),
8.66 (s, 1H), 8.47-8.45 (m, 1H), 7.76-7.73 (m, 1H), 7.51-7.45 (m,
2H), 4.37-4.32 (m, 1H), 3.12-3.07 (m, 2H), 2.84-2.73 (m, 4H), 2.53
(brs, 3H), 2.42 (brs, 1H), 2.34-2.23 (m, 2H), 2.15 (brs, 1H), 2.09
(brs, 1H), 1.92-1.83 (m, 3H), 1.52-1.49 (m, 8H), 1.36 (m, 4H),
1.26-1.23 (m, 3H), 1.14-1.10 (m, 9H), 1.03-1.00 (m, 2H), 0.90-0.89
(m, 9H), 0.81-0.79 (m, 10H); ESI-MS: m/z 799.73 (M+H).sup.+; HPLC:
99.31%.
Example 30
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(2-methylfuran-3-carboxam-
ido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a--
octadecahydro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00107##
[0465] Intermediate 1 was coupled with 2-methylfuran-3-carboxylic
acid followed by hydrolysis gave the desired product as a white
solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.14 (brs,
1H), 7.64 (s, 1H), 7.519 (d, J.sub.AB=1.5 Hz, 1H), 7.29 (s, 1H),
6.915 (d, J.sub.AB=1.5 Hz, 1H), 4.38-4.33 (m, 1H), 3.11-3.07 (m,
1H), 2.88-2.67 (m, 5H), 2.48-2.46 (m, 4H), 2.40-2.15 (m, 4H),
2.09-2.03 (m, 1H), 1.92-1.58 (m, 10H), 1.40-1.36 (m, 6H), 1.28-1.17
(m, 5H), 1.16-1.10 (m, 8H), 1.06-0.97 (m, 3H), 0.95-0.80 (m, 15H);
ESI-MS: m/z 788.73 (M+H).sup.+; HPLC: 99.08%.
Example 31
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(2-morpholinonicotinamido-
)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-oct-
adecahydro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00108##
[0467] Intermediate 1 was coupled with 2-morpholinonicotinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.09-12.06 (brs,
1H), 9.06 (s, 1H), 8.31-8.29 (m, 1H), 7.96-7.93 (m, 1H), 7.63 (s,
1H), 7.05-7.01 (m, 1H), 4.36 (m, 1H), 3.69 (brs, 4H), 3.13-3.07 (m,
6H), 2.79-2.72 (m, 4H), 2.40-2.12 (m, 6H), 1.96-1.78 (m, 5H),
1.61-1.55 (m, 11H), 1.39-1.26 (m, 10H), 1.15-1.04 (m, 12H),
0.91-0.81 (m, 9H); ESI-MS: m/z 871.78 (M+H).sup.+; HPLC:
93.74%.
Example 32
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(pyrimidine-2-carboxamido-
)propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobuta-
ne-1-carboxylic acid
##STR00109##
[0469] Intermediate 1 was coupled with pyrimidine-2-carboxylic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.12 (brs, 1H)
8.98-8.97 (m, 3H), 7.72-7.67 (m, 2H), 4.35 (m, 1H), 3.13-3.07 (m,
1H), 2.82-2.73 (m, 4H), 2.44-2.12 (m, 4H), 1.92-1.85 (m, 3H),
1.83-1.63 (m, 3H), 1.48 (m, 3H), 1.35-1.26 (m, 9H), 1.15-1.10 (m,
8H), 1.03-0.99 (m, 2H), 0.902-0.88 (m, 10H), 0.81-0.80 (m, 12H);
ESI-MS: m/z 787.62 (M+H).sup.+; HPLC: 92.90%.
Example 33
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2,5-dimethylfuran-
-3-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00110##
[0471] Intermediate 1 was coupled with
2,5-dimethylfuran-3-carboxylic acid followed by hydrolysis gave the
desired product as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 12.17 (brs, 1H), 7.51 (s, 1H), 7.27 (s,
1H), 6.49 (s, 1H), 4.38-4.32 (m, 1H), 3.11-3.07 (m, 1H), 2.87-2.73
(m, 3H), 2.44-2.38 (m, 4H), 2.31-2.22 (m, 5H), 2.09-2.03 (m, 1H),
1.92-1.83 (m, 3H), 1.75-1.58 (m, 5H), 1.38-1.37 (m, 9H), 1.30-1.26
(m, 5H), 1.13-1.10 (m, 9H), 1.02 (m, 2H), 0.95-0.81 (m, 18H);
ESI-MS: m/z 803.59 (M+H).sup.+; HPLC: 95.83%.
Example 34
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(1,1-dioxidothi-
omorpholino)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pent-
amethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-c-
arboxylic acid
##STR00111##
[0473] Intermediate 1 was coupled with
4-(carboxymethyl)thiomorpholin-4-ium 1,1-dioxide
2,2,2-trifluoroacetate (Intermediate 22) followed by hydrolysis
gave the desired product as a white solid. .sup.1H NMR (300 MHz,
DMSO-d6): .delta. ppm 12.19 (brs, 1H), 7.15 (s, 1H), 6.73 (brs,
1H), 4.38-4.33 (m, 1H), 3.11-3.07 (m, 1H), 2.86-2.73 (m, 3H),
2.39-2.27 (m, 3H), 2.17-1.99 (m, 2H), 1.92-1.87 (m, 3H), 1.69-1.56
(m, 8H), 1.39-1.27 (m, 21H), 1.14-1.09 (m, 12H), 0.91-0.81 (m,
16H); HPLC: 94.66%.
Example 35
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(piperidine-4-carboxamido-
)propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobuta-
ne-1-carboxylic acid hydrochloride
##STR00112##
[0474] Step 1: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(1-(tert-butoxycar-
bonyl)piperidine-4-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8-
,
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a--
octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclo-
butane-1-carboxylic acid
##STR00113##
[0476] Intermediate 1 was coupled with 1-(tert-butoxycarbonyl)
piperidine-4-carboxylic acid followed by hydrolysis gave the
desired product as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 12.16 (brs, 1H), 7.14 (s, 1H), 6.09 (s,
1H), 4.47-4.42 (m, 1H), 4.11 (brs, 2H), 3.15 (m, 1H), 3.02-2.57 (m,
8H), 2.28-2.22 (m, 3H), 2.05-1.83 (m, 3H), 1.67-1.62 (m, 4H),
1.56-1.54 (m, 7H), 1.45-1.40 (m, 13H), 1.36-1.34 (s, 6H), 1.28-1.20
(m, 9H), 1.13 (s, 3H), 0.96-0.91 (m, 11H), 0.85 (m, 7H); ESI-MS:
m/z 892.53 (M+H).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-3a-(2-methyl-2-(piperidine-4-carboxamido)propanamido)-
-2-oxo-3,3a,4, 5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride
[0477] To a stirred solution of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(1-(tert-butoxycar-
bonyl)piperidine-4-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,
5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,1-
3,13a-octadeca
hydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane--
1-carboxylic acid (step 1, 0.52 g, 0.58 mmol, 1.0 eq) in DCM (15
ml) then cooled to 0.degree. C. and added 1,4-Dioxane.Hcl (10 ml)
then the reaction mixture was stirred at room temperature for about
14 hours. TLC indicated starting material was consumed and the
desired product was observed. The organic phase was evaporated
under reduced pressure to get the solid compound, that solid
compound was stirred in MTBE for about 1 hour and filtered through
Buchner funnel then the solid was washed with n-pentane then the
solid compound was filtered through the Buchner funnel to obtain
the desired product (210 mg, yield: 43.75%) as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.16 (brs, 1H),
8.91-8.88 (m, 1H), 8.43-8.37 (m, 1H), 7.88 (s, 1H), 7.24 (s, 1H),
4.38-4.33 (m, 1H), 3.26-3.22 (m, 2H), 3.07 (s, 2H), 2.85-2.83 (m,
3H), 2.39-2.31 (m, 2H), 2.06-1.58 (m, 14H), 1.48-1.32 (m, 14H),
1.27-1.10 (m, 16H), 1.07-0.81 (m, 15H); ESI-MS: m/z 792.57
(M+H).sup.+; HPLC: 91.08%, Chloride content is 4.2%.
Example 36
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-2-amino-3-met-
hylbutanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid hydrochloride
##STR00114##
[0478] Step 1: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-2-((tert-buto-
xycarbonyl)amino)-3-methylbutanamido)-2-methylpropanamido)-1-isopropyl-5a,-
5b,8,8,
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,1-
3,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethy-
lcyclobutane-1-carboxylic acid
##STR00115##
[0480] Intermediate 1 was coupled with
(tert-butoxycarbonyl)-L-valine followed by hydrolysis gave the
desired product as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 12.09 (brs, 1H), 7.98-6.91 (m, 3H),
4.38-4.33 (m, 1H), 3.67-3.54 (m, 1H), 3.11-3.07 (m, 2H), 2.83-2.79
(m, 2H), 2.39-2.09 (m, 2H), 2.01-1.86 (m, 6H), 1.75-1.49 (m, 7H),
1.38-1.26 (m, 24H), 1.13-1.107 (m, 12H), 0.91-0.82 (m, 20H);
ESI-MS: m/z 880.48 (M+H).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-2-amino-3-met-
hylbutanamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl--
2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-c-
yclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxyli-
c acid hydrochloride
[0481] To a stirred solution of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-2-((tert-buto-
xycarbonyl)amino)-3-methylbutanamido)-2-methylpropanamido)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11-
a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2-
,2-dimethylcyclobutane-1-carboxylic acid (step 1, 0.52 g, 0.58
mmol, 1.0 eq) in DCM (15 ml) then cool to 0.degree. C. and added
1,4-Dioxane.Hcl (10 ml) then the reaction was stirred at room
temperature for about 14 hours. TLC indicated starting material was
consumed and the desired product was observed. The organic phase
was evaporated under reduced pressure to get the solid compound,
that solid compound was stirred in MTBE for about 1 hour and
filtered through Buchner funnel then the solid was washed with
n-pentane then the solid compound was filtered through the Buchner
funnel to obtain the desired product (55 mg, yield: 14.86%) as a
white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.08
(brs, 1H), 8.56-8.40 (m, 3H), 7.32-7.26 (m, 1H), 4.38-4.36 (m, 1H),
3.62 (brs, 1H), 3.17-3.07 (m, 2H), 2.82-2.73 (m, 3H), 2.39-2.03 (m,
5H), 1.92-1.86 (m, 3H), 1.77-1.65 (m, 7H), 1.46-1.34 (m, 12H), 1.26
(s, 4H), 1.13-1.06 (m, 12H), 0.90-0.81 (m, 19H); ESI-MS: m/z 802.68
(M+Na).sup.+; HPLC: 95.54%, Chloride content 4.65%.
Example 37
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00116##
[0483] Intermediate 3 was coupled with 4-chlorobenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d6): .delta. ppm 12.14 (s, 1H), 8.85 (s,
1H), 7.93 (d, J=6.0 Hz, 2H), 7.57 (d, J=6.0 Hz, 2H), 7.48 (s, 1H),
4.36-4.31 (m, 1H), 3.11-3.07 (m, 2H), 2.83-2.67 (m, 5H), 2.38-2.12
(m, 6H), 1.89-1.83 (m, 3H), 1.71-1.63 (m, 4H), 1.57-1.47 (m, 8H),
1.43-1.39 (m, 11H), 1.29-1.26 (m, 6H), 1.12-0.80 (m, 12H); ESI-MS:
m/z 817.45 (M+H).sup.+; HPLC: 97.69%.
Example 38
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclopropane-1-carbox
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00117##
[0485] Intermediate 3 was coupled with 6-methylnicotinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.16 (brs, 1H),
8.95-8.86 (m, 2H), 8.14-8.11 (m, 1H), 7.50-7.36 (m, 2H), 4.37-4.31
(m, 1H), 3.11-3.07 (m, 1H), 2.83-2.76 (m, 2H), 2.45-2.07 (m, 6H),
1.89-1.83 (m, 2H), 1.71-1.47 (m, 9H), 1.44-1.26 (m, 20H), 1.29-1.07
(m, 4H), 1.03-0.80 (m, 16H); ESI-MS: m/z 820.44 (M+H).sup.+ HPLC:
96.39%.
Example 39
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-fluorobenzamido-
)cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00118##
[0487] Intermediate 3 was coupled with 4-fluorobenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d6): .delta. ppm 12.14 (s, 1H),
8.81-8.00 (s, 1H), 7.98-7.96 (m, 2H), 7.49 (s, 1H), 7.36-7.30 (m,
2H), 4.36-4.31 (m, 1H), 3.17-3.07 (m, 2H), 2.83-2.76 (m, 2H),
2.45-2.12 (m, 5H), 1.89-1.83 (m, 2H), 1.73-1.57 (m, 9H), 1.47-1.43
(m, 11H), 1.26-1.12 (m, 9H), 1.10-1.03 (m, 5H), 1.00-0.90 (m, 12H);
ESI-MS: m/z 823.35 (M+Na).sup.+; HPLC: 99.10%.
Example 40
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(4-methylbenzamido)cyclopropane-1-carboxamido)-2-oxo-3-
,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00119##
[0489] Intermediate 3 was coupled with 4-methylbenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.03 (brs, 1H),
8.74 (s, 1H), 7.80 (d, J=6.0 Hz, 2H), 7.43 (s, 1H), 7.28 (d, J=6.0
Hz, 2H), 4.36-4.31 (m, 1H), 3.11-3.07 (m, 1H), 2.82-2.73 (m, 3H),
2.45 (m, 1H), 2.39-2.12 (m, 8H), 1.92-1.83 (m, 2H), 1.71-1.40 (m,
9H), 1.26 (m, 8H), 1.12-1.10 (m, 9H), 1.02-1.00 (m, 4H), 0.90-0.87
(m, 5H), 0.84-0.80 (m, 10H); ESI-MS: m/z 819.16 (M+Na).sup.+; HPLC:
98.11%.
Example 41
Preparation of
(1S,3R)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)cyclopropane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00120##
[0491] Intermediate 4 was coupled with 4-chlorobenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d6): .delta. ppm 12.14 (s, 1H), 8.85 (s,
1H), 7.93 (d, J=6.0 Hz, 2H), 7.57 (d, J=6.0 Hz, 2H), 7.48 (s, 1H),
4.36-4.31 (m, 1H), 3.11-3.07 (m, 1H), 2.87-2.73 (m, 3H), 2.45-2.38
(m, 1H), 2.33-2.22 (m, 2H), 2.18-2.12 (m, 1H), 1.98-1.82 (m, 2H),
1.73-1.40 (m, 8H), 1.29-1.26 (m, 10H), 1.17-1.10 (m, 7H), 1.01-0.93
(m, 4H), 0.89 (s, 3H) 0.84 (s, 3H) 0.79 (s, 12H); ESI-MS: m/z
817.50 (M+H).sup.+; HPLC: 93.55%.
Example 42
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)cyclobutane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,-
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopent-
a[a]chrysen-9-yl)oxy)
carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
##STR00121##
[0493] Intermediate 5 was coupled with 4-chlorobenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d6): .delta. ppm 12.14 (brs, 1H), 8.79
(s, 1H), 7.955 (d, J=9.0 Hz, 2H), 7.565 (d, J=9.0 Hz, 2H), 7.44 (s,
1H), 4.35-4.30 (m, 1H), 3.11-3.04 (m, 1H), 2.82-2.66 (m, 3H),
2.44-2.38 (m, 1H), 2.30-2.19 (m, 6H), 1.92-1.38 (m, 12H), 1.26
(brs, 8H), 1.15-1.10 (m, 7H), 0.98 (s, 3H), 0.90 (s, 3H), 0.83 (s,
3H), 0.79-0.77 (m, 6H), 0.72 (s, 3H), 0.58 (s, 3H); ESI-MS: m/z
853.62 (M+Na).sup.+; HPLC: 98.95%.
Example 43
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclobutane-1-carboxamido)-2-oxo-
-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclop-
enta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00122##
[0495] Intermediate 5 was coupled with 6-methylnicotinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.09 (brs, 1H),
8.98-8.83 (m, 2H), 8.17-8.13 (m, 1H), 7.46-7.36 (m, 2H), 4.35-4.30
(m, 1H), 3.11-3.07 (m, 1H), 2.94-2.68 (m, 4H), 2.59-2.56 (m, 2H),
2.44-2.13 (m, 6H), 1.89-1.76 (m, 13H), 1.26-1.23 (m, 8H), 1.15-1.11
(m, 7H), 0.97 (m, 3H), 0.90-0.71 (m, 15H), 0.58 (s, 3H); ESI-MS:
m/z 812.52 (M+H).sup.+; HPLC: 94.64%.
Example 44
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-2-oxo-3a-(1-(pyrimidine-2-carboxamido)cyclobutane-1-carboxam-
ido)-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cy-
clopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00123##
[0497] Intermediate 5 was coupled with pyrimidine-2-carboxylic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.12 (brs, 1H),
9.31 (s, 1H), 9.00-8.99 (m, 2H), 7.74-7.70 (m, 1H), 7.51 (s, 1H),
4.35-4.29 (m, 1H), 3.10-3.06 (m, 1H), 2.82-2.75 (m, 2H), 2.72-2.63
(m, 3H), 2.34-2.18 (m, 6H), 1.92-1.36 (m, 12H), 1.26-1.11 (m, 15H),
1.01-0.97 (m, 2H), 0.90 (brs, 3H), 0.82-0.68 (m, 12H), 0.42 (brs,
3H); ESI-MS: m/z 799.79 (M+H).sup.+; HPLC: 98.35%.
Example 45
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(2-morpholinonicotinamido)cyclobutane-1-carboxamido)-2-
-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cy-
clopenta[a]
chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00124##
[0499] Intermediate 5 was coupled with 2-morpholinonicotinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.15 (brs, 1H),
8.94 (s, 1H), 8.27-8.25 (m, 1H), 7.86-7.83 (m, 1H), 7.48 (s, 1H),
6.95-6.91 (m, 1H), 4.37-4.32 (m, 1H), 3.67 (brs, 4H), 3.25 (bs,
2H), 3.11-3.07 (m, 2H), 2.79-2.73 (m, 3H), 2.50 (m, 2H), 2.41-2.14
(m, 6H), 1.91-1.78 (m, 7H), 1.57-1.46 (m, 6H), 1.31-1.26 (m, 6H),
1.14-1.10 (m, 9H), 0.90-0.88 (m, 10H), 0.81-0.80 (m, 10H); ESI-MS:
m/z 905.84 (M+Na).sup.+; HPLC: 97.06%.
Example 46
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclopentane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo--
3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclope-
nta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00125##
[0501] Intermediate 6 was coupled with 4-chlorobenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.08 (brs, 1H),
8.31 (s, 1H), 7.93 (d, J=9.0 Hz, 2H), 7.54 (d, J=9.0 Hz, 2H), 7.43
(s, 1H), 4.36-4.30 (m, 1H), 3.10-3.07 (m, 1H), 2.79 (m, 3H),
2.38-2.23 (m, 3H), 2.16-2.08 (m, 3H), 1.95-1.82 (m, 4H), 1.76-1.57
(m, 8H), 1.43-1.39 (m, 2H), 1.26 (brs, 8H), 1.03-1.10 (m, 8H),
1.04-0.96 (m, 2H), 0.90 (brs, 3H), 0.83-0.76 (m, 12H), 0.69 (brs,
3H); ESI-MS: m/z 845.21 (M+H).sup.+; HPLC: 95.88%.
Example 47
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclopentane-1-carboxamido)-2-ox-
o-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclo-
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00126##
[0503] Intermediate 6 was coupled with 6-methylnicotinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.16 (brs, 1H),
8.95 (s, 1H), 8.34 (s, 1H), 8.15-8.11 (m, 1H), 7.43 (s, 1H),
7.37-7.34 (m, 1H), 4.36-4.31 (m, 1H), 3.11-3.06 (m, 1H), 2.79-2.76
(m, 1H), 2.39 (m, 1H), 2.16-2.07 (m, 4H), 1.95-1.43 (m, 16H),
1.39-1.25 (m, 11H), 1.14-1.12 (m, 7H), 1.04-0.99 (m, 2H), 0.96-0.86
(m, 18H), 0.84 (brs, 3H); ESI-MS: m/z 848.44 (M+Na).sup.+; HPLC:
99.78%.
Example 48
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(1-(4-chlorobenzamido-
)
cyclohexane-1-carboxamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3-
,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00127##
[0505] Intermediate 7 was coupled with 4-chlorobenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.17 (brs, 1H),
7.90 (d, J=8.4 Hz, 2H), 7.87 (s, 1H), 7.55 (d, J=8.4 Hz, 2H), 7.30
(s, 1H), 4.37-4.31 (m, 1H), 3.11-3.07 (m, 1H), 2.82-2.76 (m, 3H),
2.31-2.06 (m, 6H), 1.89-1.71 (m, 8H), 1.63-1.39 (s, 10H), 1.31 (m,
10H), 1.26-1.10 (m, 9H), 1.01-0.90 (m, 2H), 0.85-0.80 (m, 14H);
ESI-MS: m/z 859.30 (M+H).sup.+; HPLC: 98.57%.
Example 49
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(6-methylnicotinamido)cyclohexane-1-carboxamido)-2-oxo-
-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclop-
enta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00128##
[0507] Intermediate 7 was coupled with 6-methylnicotinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.16 (brs, 1H),
8.926-8.920 (m, 1H), 8.12-8.09 (m, 1H), 7.93 (s, 1H), 7.37-7.31 (m,
2H), 4.37-4.31 (m, 1H), 3.07-2.75 (brs, 3H), 2.41-2.35 (m, 2H),
2.21-2.06 (m, 2H), 1.89-1.75 (m, 5H), 1.52 (brs, 10H), 1.42-1.26
(m, 8H), 1.13-1.10 (m, 10H), 1.06-0.96 (m, 4H), 0.90-0.79 (m, 22H);
ESI-MS: m/z 840.52 (M+H).sup.+; HPLC: 99.89%.
Example 50
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(1-(4-methylbenzamido)cyclohexane-1-carboxamido)-2-oxo-3,-
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopent-
a[a]chrysen 9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00129##
[0509] Intermediate 7 was coupled with 4-methylbenzoic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.18 (brs, 1H),
8.74 (s, 1H), 7.757 (d, J=13.2 Hz, 2H), 7.34 (s, 1H), 7.291 (d,
J=13.2, Hz, 2H), 4.37-4.32 (m, 1H), 3.11-3.06 (m, 1H), 2.82-2.76
(m, 1H), 2.43-2.36 (m, 6H), 2.16-2.06 (m, 6H), 1.89-1.64 (m, 7H),
1.51 (brs, 9H), 1.32-1.26 (m, 10H), 1.13-1.11 (m, 8H), 1.06-1.02
(m, 4H), 0.90-0.86 (m, 14H); ESI-MS: m/z 839.46 (M+H).sup.+; HPLC:
92.98%.
Example 51
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(methylsulfonyl)benzamido)
propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobuta-
ne-1-carboxylic acid
##STR00130##
[0510] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,
8,11a-pentamethyl-3a-(2-methyl-2-(4-(methylsulfonyl)benzamido)propanamido-
)-2-oxo-3,3a,
4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00131##
[0512] To a stirred solution of 4-(methylsulfonyl)benzoic acid
(0.778 g, 3.889 mmol, 3.0 eq) in DMF (10 ml) was added HBTU (1.475
g, 3.889 mmol, 3.0 eq) followed by triethylamine (1.08 ml, 7.776
mmol, 6.0 eq). The reaction mixture was stirred at room temperature
for about 30 minutes then
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methyl
propanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,11,
11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-di methylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 1.0 g, 1.296 mmol, 1.0 eq) was added and stirred
at room temperature for overnight. TLC indicated starting material
was consumed and the desired product was observed. The reaction
mixture was diluted with water (120 ml) and extracted with ethyl
acetate (2.times.50 ml). The combined organic extracts were washed
with water (70 ml), dried over Na.sub.2SO.sub.4, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-2% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the desired product (0.75 g, yield: 60.97%) as a white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 8.0 (d, J=6.9
Hz, 2H), 7.96 (d, J=6.9 Hz, 2H), 7.34 (m, 5H), 7.18 (s, 1H), 6.70
(s, 1H), 5.15, 5.09 (ABq, J.sub.AB=12.3 Hz, 2H), 4.45 (dd, J=11.1,
4.5 Hz, 1H), 3.22-3.11 (m, 1H), 3.06 (s, 3H), 2.84-2.72 (m, 3H),
2.70-2.60 (m, 2H), 2.38-2.26 (m, 2H), 2.09-1.98 (m, 1H), 1.97-1.83
(m, 3H), 1.72 (brs, 6H), 1.71-1.40 (m, 9H), 1.40-1.29 (m, 2H), 1.34
(s, 3H), 1.27-1.17 (m, 8H), 1.08 (s, 3H), 0.96 (s, 3H), 0.95 (s,
3H), 0.89 (s, 3H), 0.85 (s, 3H), 0.84 (s, 3H), 0.79 (m, 1H);
ESI-MS: m/z 976.15 (M+Na).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-3a-(2-methyl-2-(4-(methylsulfonyl)benzamido)propanami-
do)-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
[0513] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11a-pentamet-
hyl-3a-(2-methyl-2-(4-(methyl sulfonyl)benzamido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 1, 0.750 g,
0.786 mmol, 1.0 eq) in MeOH (7.5 ml) and THF (7.5 ml) was added
aqueous 2.5N KOH solution (2.36 ml, 5.89 mmol, 7.5 eq). The
reaction mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (10 ml),
cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (2.times.50 ml). The combined organic extracts
were washed with water (30 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-5% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the solid. To this solid compound, CH.sub.3CN: EtOAc (4:1,
10 ml) was added and heated to reflux for about 30 minutes. The
mixture was cooled to 0.degree. C., solid was filtered and dried
under vacuum to obtain the desired product (0.275 g, yield: 40.48%)
as a white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm
8.02 (d, J=8.4 Hz, 2H), 7.96 (d, J=8.4 Hz, 2H), 7.10 (s, 1H), 6.63
(s, 1H), 4.46 (dd, J=11.1, 4.5 Hz, 1H), 3.22-3.11 (m, 1H), 3.06 (s,
3H), 2.86-2.75 (m, 3H), 2.71-2.52 (m, 2H), 2.40-2.26 (m, 2H),
2.10-2.01 (m, 1H), 2.0-1.78 (m, 3H), 1.72 (brs, 6H), 1.69-1.40 (m,
9H), 1.40-1.32 (m, 2H), 1.37 (s, 3H), 1.26-1.20 (m, 8H), 1.08 (s,
3H), 1.06 (s, 3H), 0.96 (s, 3H), 0.89 (s, 3H), 0.86 (s, 3H), 0.85
(s, 3H), 0.79 (m, 1H); ESI-MS: m/z 863.6 (M+H).sup.+; HPLC:
97.9%.
[0514] The below examples 52-58 were prepared by the procedure
similar (including reagents and reaction conditions) to the above
described in the synthesis of example-51 using with their
appropriate intermediates.
Example 52
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((S)-piperidine-3-carboxamido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid hydrochloride
##STR00132##
[0515] Step 1: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-1-(tert-butox-
ycarbonyl)piperidine-3-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b-
,8,8,
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,-
13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylc-
yclobutane-1-carboxylic acid
##STR00133##
[0517] Intermediate 1 was coupled with
(S)-1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid followed by
hydrolysis gave the desired product as a white solid. .sup.1H NMR
(300 MHz, CDCl.sub.3): .delta. ppm 4.47 (dd, J=11.1, 4.5 Hz, 1H),
3.83-3.50 (m, 2H), 3.40-3.02 (m, 3H), 3.0-2.73 (m, 4H), 2.68-2.53
(m, 2H), 2.38-2.20 (m, 4H), 2.10-1.65 (m, 15H), 1.58-1.53 (m, 4H),
1.53 (s, 3H), 1.46 (s, 9H), 1.37 (m, 4H), 1.35-1.32 (m, 2H),
1.25-1.18 (m, 7H), 1.14 (s, 3H), 1.07 (s, 3H), 0.93 (s, 3H), 0.92
(s, 3H), 0.87 (s, 3H), 0.85 (s, 3H), 0.83 (m, 1H); ESI-MS: m/z
914.71 (M+Na).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-3a-(2-methyl-2-((S)-piperidine-3-carboxamido)propanam-
ido)-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid hydrochloride
[0518] A solution of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-1-(tert-butox-
ycarbonyl)piperidine-3-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b-
,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,1-
3a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcy-
clobutane-1-carboxylic acid (step 1, 0.500 g, 0.560 mmol, 1.0 eq)
and 3N HCl in 1,4-dioxane (10 ml) was stirred at room temperature
for overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was evaporated
under reduced pressure, MTBE (10 ml) was added and heated to reflux
for about 30 minutes. The reaction mixture was cooled to 0.degree.
C., filtered, solid was washed with MTBE (10 ml) and dried under
vacuum to obtain the desired product (0.220 g, yield: 47.4%) as a
white solid. .sup.1H NMR (300 MHz, CD.sub.3OD): .delta. ppm 8.02
(s, 1H), 7.15 (s, 1H), 4.32-4.23 (m, 1H), 3.07-2.85 (m, 6H),
2.80-2.55 (m, 4H), 2.50-2.21 (m, 3H), 2.21-1.93 (m, 2H), 1.93-1.76
(m, 4H), 1.76-1.58 (m, 5H), 1.58-1.40 (m, 4H), 1.40-1.36 (m, 2H),
1.36-1.21 (m, 11H), 1.17 (s, 3H), 1.15-1.10 (m, 1H), 1.08-1.0 (m,
9H), 0.96-0.89 (m, 1H), 0.87-0.78 (m, 7H), 0.72 (brs, 6H); ESI-MS:
m/z 792.58 (M-HCl+H).sup.+; HPLC: 90.7%; chloride content by Ion
chromatography: 6.8%.
Example 53
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(4-chlorophenyl-
)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-o-
xo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cycl-
openta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00134##
[0520] Intermediate 1 was coupled with 2-(4-chlorophenyl)acetic
acid followed by hydrolysis gave the desired product as a white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.33 (d,
J=8.1 Hz, 2H), 7.19 (d, J=8.4 Hz, 2H), 7.11 (brs, 1H), 5.84 (brs,
1H), 4.47 (dd, J=11.1, 4.5 Hz, 1H), 3.50 (s, 2H), 3.20-3.08 (m,
1H), 2.87-2.70 (m, 3H), 2.67-2.53 (m, 2H), 2.29-2.02 (m, 3H),
1.98-1.65 (m, 6H), 1.55-1.47 (m, 4H), 1.51 (s, 3H), 1.49 (s, 3H),
1.42-1.35 (m, 2H), 1.37 (s, 3H), 1.28-1.18 (m, 9H), 1.13-1.03 (m,
1H), 1.10 (s, 3H), 1.07 (s, 3H), 0.93 (s, 3H), 0.91 (s, 3H),
0.89-0.78 (m, 7H); ESI-MS: m/z 855.6 (M+Na).sup.+; HPLC: 89.2%+9%
isomer.
Example 54
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(pyrazolo[1,5-a]pyrimidine-3-carboxamido)prop-
anamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecah-
ydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1--
carboxylic acid
##STR00135##
[0522] Intermediate 1 was coupled with
pyrazolo[1,5-a]pyrimidine-3-carboxylic acid followed by hydrolysis
gave the desired product as an off-white solid. .sup.1H NMR (300
MHz, CDCl.sub.3): .delta. ppm 11.74 (brs, 1H), 9.02 (dd, J=7.2, 1.5
Hz, 1H), 8.75 (dd, J=4.2, 1.5 Hz, 1H), 8.57 (s, 1H), 8.27 (s, 1H),
7.66 (m, 1H), 7.16 (dd, J=6.9, 4.2 Hz, 1H), 4.40 (dd, J=10.5, 4.5
Hz, 1H), 3.20-3.10 (m, 1H), 2.97-2.83 (m, 1H), 2.81-2.70 (m, 2H),
2.62-2.58 (m, 2H), 2.42-2.12 (m, 2H), 2.07-1.87 (m, 4H), 1.80-1.50
(m, 6H), 1.66 (s, 3H), 1.62 (s, 3H), 1.47-1.31 (m, 4H), 1.34 (s,
3H), 1.23-1.15 (m, 8H), 1.05-0.97 (m, 1H), 1.02 (s, 3H), 0.99 (s,
3H), 0.92 (s, 3H), 0.88-0.78 (m, 10H); ESI-MS: m/z 826.7
(M+H).sup.+; HPLC: 91%.
Example 55
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-aminothiazole-4-
-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b. 8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00136##
[0524] Intermediate 1 was coupled with 2-aminothiazole-4-carboxylic
acid (Intermediate 14) followed by hydrolysis gave the desired
product as an off-white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6):
.delta. ppm 12.18 (brs, 1H), 7.95 (s, 1H), 7.66 (s, 1H), 7.20 (brs,
2H), 7.13 (s, 1H), 4.40-4.32 (m, 1H), 3.18-3.07 (m, 1H), 2.85-2.72
(m, 3H), 2.40-2.22 (m, 3H), 2.13 (d, J=18.3 Hz, 1H), 2.0-1.82 (m,
3H), 1.80-1.61 (m, 3H), 1.60-1.44 (m, 2H), 1.52 (brs, 6H),
1.43-1.30 (m, 5H), 1.30-1.23 (m, 2H), 1.26 (s, 3H), 1.18-1.10 (m,
7H), 1.08-0.98 (m, 1H), 1.0 (s, 3H), 0.91 (s, 3H), 0.89 (s, 3H),
0.88-0.72 (m, 10H); ESI-MS: m/z 829.54 (M+Na).sup.+; HPLC:
91.3%.
Example 56
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(4-(5-methyl-1,3,4-oxadiazol-2-yl)
benzamido)propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,1-
3,13a-octa
decahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimeth-
ylcyclobutane-1-carboxylic acid
##STR00137##
[0526] Intermediate 1 was coupled with
4-(5-methyl-1,3,4-oxadiazol-2-yl)benzoic acid (Intermediate 15)
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 8.11 (d, J=8.4 Hz,
2H), 7.91 (d, J=8.4 Hz, 2H), 6.98 (s, 1H), 6.92 (s, 1H), 4.46 (dd,
J=11.1, 4.5 Hz, 1H), 3.22-3.10 (m, 1H), 2.88-2.70 (m, 4H), 2.65 (s,
3H), 2.61-2.52 (m, 1H), 2.38-2.27 (m, 2H), 2.10-1.83 (m, 5H),
1.80-1.67 (m, 1H), 1.72 (s, 3H), 1.71 (s, 3H), 1.65-1.42 (m, 7H),
1.40-1.31 (m, 2H), 1.36 (s, 3H), 1.28-1.20 (m, 7H), 1.08-1.04 (m,
7H), 0.95 (s, 3H), 0.90-0.78 (m, 10H); ESI-MS: m/z 889.67
(M+Na).sup.+; HPLC: 93.3%.
Example 57
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-(1,1-dioxidothi-
omorpholino)benzamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pent-
a
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecah-
ydro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00138##
[0528] Intermediate 1 was coupled with
4-(1,1-dioxidothiomorpholino)benzoic acid (Intermediate 16)
followed by hydrolysis gave the desired product as an off-white
solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.10 (s,
1H), 7.92 (s, 1H), 7.80 (d, J=8.4 Hz, 2H), 7.31 (s, 1H), 7.07 (d,
J=8.7 Hz, 2H), 4.40-4.30 (m, 1H), 3.90 (m, 4H), 3.09 (m, 5H),
2.87-2.70 (m, 3H), 2.48-2.18 (m, 4H), 2.06 (d, J=18.3 Hz, 1H),
1.98-1.80 (m, 3H), 1.80-1.48 (m, 6H), 1.42 (s, 3H), 1.40 (s, 3H),
1.38-1.20 (m, 6H), 1.26 (s, 3H), 1.18-1.10 (m, 6H), 0.93-0.75 (m,
20H); ESI-MS: m/z 940.72 (M+Na).sup.+; HPLC: 92.4%.
Example 58
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(4-((1,1-dioxidoth-
iomorpholino)methyl)benzamido)-2-methylpropanamido)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,1-
3a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcy-
clobutane-1-carboxylic acid
##STR00139##
[0530] Intermediate 1 was coupled with
4-((1,1-dioxidothiomorpholino)methyl)benzoic acid (Intermediate 17)
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 11.8 (brs, 1H),
8.13 (s, 1H), 7.86 (d, J=7.8 Hz, 2H), 7.42 (d, J=7.8 Hz, 2H), 7.36
(s, 1H), 4.38-4.31 (m, 1H), 3.73 (s, 2H), 3.11 (m, 5H), 2.92-2.70
(m, 7H), 2.48-2.18 (m, 3H), 2.12-1.98 (m, 2H), 1.96-1.80 (m, 3H),
1.80-1.51 (m, 5H), 1.50-1.38 (m, 1H), 1.43 (s, 3H), 1.41 (s, 3H),
1.38-1.24 (m, 4H), 1.26 (s, 3H), 1.20-1.06 (m, 8H), 1.05-0.96 (m,
2H), 0.90 (s, 3H), 0.86 (m, 6H), 0.79 (m, 9H); ESI-MS: m/z 954.75
(M+Na).sup.+; HPLC: 92.9%.
Example 59
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(dimethylamino)-
acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-ox-
o-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclo-
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00140##
[0531] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(dimethyl
amino)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentameth-
yl-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00141##
[0533] To a stirred solution of dimethyl glycine hydrochloride
(0.543 g, 3.893 mmol, 3.0 eq) in DMF (10 ml) was added HATU (0.980
g, 2.59 mmol, 2.0 eq) followed by DIPEA (1.34 ml, 7.78 mmol, 6.0
eq). The reaction mixture was stirred at room temperature for about
30 minutes then
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropan
amido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,-
8,9,10,11,11a,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl) 3-benzyl
(1S,3R)-2,2-dimethyl cyclobutane-1,3-dicarboxylate (Intermediate 1,
1.0 g, 1.297 mmol, 1.0 eq) was added and stirred at the same
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was diluted with water (150 ml) and extracted with DCM (3.times.20
ml). The combined organic extracts were washed with water (20 ml),
dried over Na.sub.2SO.sub.4, filtered and evaporated under reduced
pressure. The residue was purified by silicagel column
chromatography by using 0-3% MeOH in DCM gradient. The fractions
containing the product were combined and concentrated under reduced
pressure to give the desired product (0.5 g, yield: 45.4%) as a
white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.99
(s, 1H), 7.46 (s, 1H), 7.35 (m, 5H), 5.15, 5.09 (ABq, J.sub.AB=12.3
Hz, 2H), 4.45 (dd, J=11.4, 4.8 Hz, 1H), 3.20-3.10 (m, 1H), 2.97 (s,
2H), 2.87-2.73 (m, 3H), 2.70-2.60 (m, 2H), 2.33 (s, 6H), 2.30-2.22
(m, 2H), 2.08-1.99 (m, 2H), 1.98-1.83 (m, 2H), 1.80-1.72 (m, 2H),
1.60-1.53 (m, 2H), 1.56 (s, 3H), 1.51 (s, 3H), 1.49-1.38 (m, 4H),
1.38-1.30 (m, 2H), 1.34 (s, 3H), 1.27-1.18 (m, 8H), 1.13 (s, 3H),
1.10-0.98 (m, 1H), 0.96 (s, 3H), 0.93 (s, 3H), 0.92 (s, 3H), 0.85
(s, 3H), 0.84 (s, 3H), 0.78 (m, 1H); ESI-MS: m/z 856.4
(M+H).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(dimethyl
amino)acetamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentameth-
yl-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
[0534] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(2-(dimethylamino)acetamido)-
-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5-
,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step
1, 0.5 g, 0.584 mmol, 1.0 eq) in MeOH (10 ml) and THF (10 ml) was
added aqueous 2.5N KOH solution (1.85 ml). The reaction mixture was
stirred at room temperature for overnight. TLC indicated starting
material was consumed and the desired product was observed. The
organic phase was evaporated under reduced pressure, the reaction
mixture was diluted with water (15 ml), cooled to 0.degree. C.,
acidified with 1N HCl to pH 5.0 and extracted with DCM (3.times.25
ml). The combined organic extracts were washed with water (10 ml),
dried over sodium sulfate, filtered and evaporated under reduced
pressure. The residue was purified by silicagel column
chromatography by using 0-10% MeOH in DCM gradient. The fractions
containing the product were combined and concentrated under reduced
pressure to give a solid. To this solid compound, acetonitrile (15
ml) was added and heated to reflux for about 30 minutes. The
mixture was cooled to 0.degree. C., the solids formed were
collected by filtration and were washed with n-hexane (10 ml) and
dried under vacuum to obtain the desired product (0.266 g, yield:
59.5%) as a white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta.
ppm 8.01 (s, 1H), 7.53 (s, 1H), 4.47 (dd, J=11.1, 4.5 Hz, 1H),
3.20-3.10 (m, 1H), 2.96 (s, 2H), 2.86-2.75 (m, 3H), 2.68-2.40 (m,
2H), 2.32 (s, 6H), 2.30-2.22 (m, 2H), 2.12-2.0 (m, 1H), 1.99-1.83
(m, 3H), 1.82-1.67 (m, 2H), 1.65-1.58 (m, 2H), 1.56 (s, 3H), 1.51
(s, 3H), 1.49-1.40 (m, 3H), 1.40-1.30 (m, 3H), 1.37 (s, 3H),
1.30-1.15 (m, 8H), 1.13 (s, 3H), 1.07 (s, 3H), 1.01 (m, 1H), 0.93
(s, 3H), 0.91 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H), 0.80 (m, 1H);
ESI-MS: m/z 766.49 (M+H).sup.+; HPLC: 97.5%.
[0535] The below examples 60-61 were prepared by the procedure
similar (including reagents and reaction conditions) to the above
described in the synthesis of example-59 using with their
appropriate intermediates.
Example 60
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(6-methylpicolinamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00142##
[0537] Intermediate 1 was coupled with 6-methylpicolinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.13 (brs, 1H),
8.76 (s, 1H), 7.90-7.80 (m, 2H), 7.62 (s, 1H), 7.47 (d, J=6.9 Hz,
1H), 4.39-4.31 (m, 1H), 3.18-3.06 (m, 1H), 2.88-2.70 (m, 3H), 2.55
(s, 3H), 2.48-2.38 (m, 2H), 2.27-2.22 (m, 2H), 2.14 (d, J=18.3 Hz,
1H), 2.0-1.62 (m, 6H), 1.55 (brs, 6H), 1.62-1.50 (m, 3H), 1.50-1.38
(m, 2H), 1.38-1.20 (m, 2H), 1.33 (s, 3H), 1.20-0.99 (m, 9H),
0.98-0.80 (m, 19H); ESI-MS: m/z 800.79 (M+H).sup.+; HPLC:
96.27%.
Example 61
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(6-methylnicotinamido)propanamido)-2-oxo-3,3a-
,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00143##
[0539] Intermediate 1 was coupled with 6-methylnicotinic acid
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 8.90 (d, J=1.8 Hz,
1H), 7.99 (dd, J=8.1, 2.4 Hz, 1H), 7.26 (d, 1H), 6.98 (brs, 1H),
6.80 (brs, 1H), 4.48 (dd, J=11.1, 4.5 Hz, 1H), 3.23-3.12 (m, 1H),
2.88-2.70 (m, 4H), 2.64 (s, 3H), 2.63-2.57 (m, 1H), 2.38-2.27 (m,
2H), 2.13-2.02 (m, 1H), 2.01-1.76 (m, 5H), 1.71 (brs, 6H),
1.66-1.43 (m, 8H), 1.39 (s, 3H), 1.37-1.20 (m, 8H), 1.20-1.15 (m,
1H), 1.08 (s, 3H), 1.07 (s, 3H), 0.96 (s, 3H), 0.90 (s, 3H), 0.88
(s, 3H), 0.86 (s, 3H), 0.80 (m, 1H); ESI-MS: m/z 800.6 (M+H).sup.+;
HPLC: 95.4%.
Example 62
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-pivalamidopropanamido)-2-oxo-3,3a,4,5,5a,5b,6-
,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9--
yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
##STR00144##
[0540] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b, 8,
8,11a-pentamethyl-3a-(2-methyl-2-pivalamidopropanamido)-2-oxo-3,3a,4,5,5a-
,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
(1R,3S)-2,2-di methylcyclobutane-1,3-dicarboxylate
##STR00145##
[0542] To a stirred solution of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 0.800 g, 1.037 mmol, 1.0 eq) in DCM (10 ml) at
0.degree. C. was added triethyl amine (0.72 ml, 5.187 mmol, 5.0 eq)
followed by pivaloyl chloride (0.192 ml, 1.556 mmol, 1.5 eq). The
reaction mixture was allowed to stir at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was diluted with
water (50 ml) and extracted with DCM (3.times.30 ml). The combined
organic extracts were washed with 0.5N HCl (10 ml), water (20 ml),
dried over sodium sulfate, filtered and evaporated under reduced
pressure. The residue was purified by silicagel column
chromatography by using 0-3% methanol in dichloromethane gradient.
The fractions containing the expected product were combined and
concentrated under reduced pressure to obtain the desired product
(0.800 g, yield: 90.19%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 7.35 (m, 5H), 5.97 (s, 1H), 5.15, 5.09
(ABq, J.sub.AB=12.3 Hz, 2H), 4.45 (dd, J=11.4, 4.8 Hz, 1H),
3.20-3.10 (m, 1H), 2.87-2.73 (m, 3H), 2.71-2.57 (m, 2H), 2.36-2.22
(m, 2H), 2.09-2.0 (m, 1H), 1.98-1.82 (m, 3H), 1.78-1.60 (m, 5H),
1.55 (s, 3H), 1.53 (s, 3H), 1.50-1.38 (m, 4H), 1.37-1.30 (m, 3H),
1.34 (s, 3H), 1.26-1.20 (m, 6H), 1.18 (s, 9H), 1.14 (s, 3H),
1.10-1.0 (m, 1H), 0.96 (s, 3H), 0.95-0.89 (m, 1H), 0.93 (s, 3H),
0.91 (s, 3H), 0.85 (s, 3H), 0.84 (s, 3H), 0.81-0.77 (m, 1H);
ESI-MS: m/z 855.56 (M+H).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-3a-(2-methyl-2-pivalamidopropanamido)-2-oxo-3,3a,4,5,-
5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
[0543] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-pivalamidopropanamido)-2--
oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step
1, 0.800 g, 0.935 mmol, 1.0 eq) in MeOH (8 ml) and THF (8 ml) was
added aqueous 2.5N KOH solution (2.8 ml, 7.015 mmol, 7.5 eq). The
reaction mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (10 ml),
cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (3.times.40 ml). The combined organic extracts
were washed with water (40 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-4% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the desired product (0.320 g, yield: 44.71%) as a white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.36 (s, 1H),
5.97 (s, 1H), 4.47 (dd, J=8.7, 3.3 Hz, 1H), 3.20-3.11 (m, 1H),
2.87-2.76 (m, 3H), 2.65-2.57 (m, 2H), 2.34-2.23 (m, 2H), 2.10-2.02
(m, 1H), 1.97-1.82 (m, 3H), 1.80-1.68 (m, 2H), 1.64-1.47 (m, 6H),
1.56 (s, 3H), 1.53 (s, 3H), 1.46-1.30 (m, 4H), 1.37 (s, 3H), 1.22
(s, 3H), 1.21 (s, 3H), 1.18 (s, 9H), 1.15 (s, 3H), 1.07 (s, 3H),
1.06-1.0 (m, 1H), 0.93 (s, 3H), 0.92 (s, 3H), 0.87 (s, 3H), 0.86
(s, 3H), 0.84-0.79 (m, 1H); ESI-MS: m/z 765.63 (M+H).sup.+; HPLC:
95.2%.
[0544] The below examples 63-66 were prepared by the procedure
similar (including reagents and reaction conditions) to the above
described in the synthesis of example-62 using with their
appropriate intermediates.
Example 63
Preparation of sodium
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(methylsulfonamido)propan-
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahyd-
ro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-ca-
rboxylate
##STR00146##
[0545] Step 1: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-3a-(2-methyl-2-(methylsulfonamido)propanamido)-2-oxo--
3,3a,4,5, 5a,
5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrys-
en-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00147##
[0547] Intermediate 1 was coupled with methanesulfonyl chloride
followed by hydrolysis gave the desired product as an off-white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 6.50 (s, 1H),
4.96 (s, 1H), 4.46 (dd, J=11.1, 4.5 Hz, 1H), 3.22-3.10 (m, 1H),
3.07 (s, 3H), 2.87-2.75 (m, 3H), 2.69-2.53 (m, 2H), 2.35-2.24 (m,
2H), 2.11-2.0 (m, 1H), 1.97-1.82 (m, 3H), 1.79-1.65 (m, 2H),
1.65-1.53 (m, 2H), 1.60 (s, 3H), 1.58 (s, 3H), 1.52-1.43 (m, 3H),
1.42-1.30 (m, 3H), 1.37 (s, 3H), 1.29-1.18 (m, 8H), 1.14 (s, 3H),
1.07 (s, 3H), 1.03-0.97 (m, 1H), 0.94 (s, 3H), 0.92 (s, 3H), 0.86
(s, 3H), 0.85 (s, 3H), 0.79 (m, 1H); ESI-MS: m/z 759.51
(M+H).sup.+.
Step 2: Synthesis of sodium
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(methylsulfonamido)propanamido)-2-oxo-3,3a,4,
5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a-
]chrysen-9-yl)
oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylate
[0548] To a stirred solution of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(methylsulfonamido)propanamido)-2-oxo-3,3a,4,-
5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]c-
hrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
(step 1, 0.320 g, 0.421 mmol, 1.0 eq) in MeOH (3.2 ml) and water
(0.15 ml) was added sodium hydroxide (0.0168 g, 0.421 mmol, 1.0
eq). The reaction mixture was stirred at room temperature for about
2 hours and then distilled out 90% of methanol under reduced
pressure. Ethyl acetate (5 ml) was added and the reaction mixture
was stirred at room temperature for overnight. The reaction mixture
was filtered, solid was washed ethyl acetate (5 ml) and then dried
under vacuum to obtain the desired product (0.185 g, yield: 56.19%)
as an off-white solid. .sup.1H NMR (300 MHz, CD.sub.3OD): .delta.
ppm 4.47-4.40 (m, 1H), 3.28-3.20 (m, 1H), 3.03 (s, 3H), 2.98-2.92
(m, 1H), 2.72-2.43 (m, 4H), 2.36-2.17 (m, 2H), 2.12-1.85 (m, 5H),
1.83-1.76 (m, 1H), 1.73-1.55 (m, 4H), 1.51 (s, 3H), 1.55-1.38 (m,
3H), 1.49 (s, 3H), 1.38-1.26 (m, 2H), 1.33 (s, 3H), 1.24-1.05 (m,
8H), 1.03 (s, 3H), 1.0 (s, 3H), 0.96 (s, 3H), 0.95-0.85 (m, 10H);
ESI-MS: m/z 781.46 (M+H).sup.+; HPLC: 97.84%; sodium content by Ion
chromatography: 3.7%.
Example 64
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((ethoxycarbonyl)a-
mino)-2-methylpropanamido)-1-isopropyl-5a,5b.
8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,1-
3a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcy-
clobutane-1-carboxylic acid
##STR00148##
[0550] Intermediate 1 was coupled with ethyl carbonochloridate
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 5.48 (s, 1H),
4.50-4.40 (m, 1H), 3.23-3.12 (m, 1H), 2.88-2.76 (m, 2H), 2.67-2.32
(m, 4H), 2.11-1.92 (m, 3H), 1.80-1.50 (m, 11H), 1.50-1.31 (m, 15H),
1.30-1.18 (m, 8H), 1.12-0.98 (m, 1H), 1.07 (s, 3H), 1.03 (s, 3H),
0.99 (s, 3H), 0.89 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H), 0.79 (m,
1H); ESI-MS: m/z 775.47 (M+Na).sup.+; HPLC: 93.5%.
Example 65
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((4-chlorophenyl)s-
ulfonamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-o-
xo-3,3a,4,5,5a,5b,6,7,7a,8,91,111,11a,11b,12,13,13a-octadecahydro-2H-cyclo-
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00149##
[0552] Intermediate 1 was coupled with 4-chlorobenzenesulfonyl
chloride followed by hydrolysis gave the desired product as an
off-white solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm
7.81 (d, J=8.7 Hz, 2H), 7.49 (d, J=8.7 Hz, 2H), 6.58 (brs, 1H),
5.40 (s, 1H), 4.47 (dd, J=11.1, 4.5 Hz, 1H), 3.22-3.10 (m, 1H),
2.88-2.75 (m, 3H), 2.67-2.53 (m, 2H), 2.40-2.26 (m, 2H), 2.11-2.0
(m, 1H), 2.0-1.82 (m, 3H), 1.80-1.68 (m, 3H), 1.68-1.48 (m, 6H),
1.47-1.30 (m, 11H), 1.27-1.20 (m, 7H), 1.15 (s, 3H), 1.07 (s, 3H),
0.97 (s, 3H), 1.05-0.98 (m, 1H), 0.97 (s, 3H), 0.95 (s, 3H), 0.87
(s, 3H), 0.86 (s, 3H), 0.83-0.80 (m, 1H); ESI-MS: m/z 855.6
(M+H).sup.+; HPLC: 99.6%.
Example 66
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(cyclo
hexanecarboxamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentame-
thyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-
-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carb-
oxylic acid
##STR00150##
[0554] Intermediate 1 was coupled with cyclohexane carbonyl
chloride followed by hydrolysis gave the desired product as a white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.51 (s, 1H),
5.78 (s, 1H), 4.47 (dd, J=11.1, 4.8 Hz, 1H), 3.20-3.09 (m, 1H),
2.87-2.77 (m, 3H), 2.65-2.53 (m, 2H), 2.32-2.20 (m, 2H), 2.12-2.0
(m, 2H), 2.0-1.90 (m, 2H), 1.88-1.75 (m, 5H), 1.75-1.65 (m, 4H),
1.56 (s, 3H), 1.52 (s, 3H), 1.55-1.50 (m, 1H), 1.50-1.40 (m, 4H),
1.40-1.37 (m, 2H), 1.37 (s, 3H), 1.36-1.28 (m, 4H), 1.27-1.18 (m,
9H), 1.14 (s, 3H), 1.07 (m, 4H), 0.93 (s, 3H), 0.91 (s, 3H), 0.87
(s, 3H), 0.85 (s, 3H), 0.80 (m, 1H); ESI-MS: m/z 813.48
(M+Na).sup.+; HPLC: 96.4%
Example 67
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((pyridin-2-ylmethyl)amino)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00151##
[0555] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b, 8,
8,11a-pentamethyl-3a-(2-methyl-2-((pyridin-2-ylmethyl)amino)propanamido)--
2-oxo-3,3a,4, 5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00152##
[0557] To a stirred solution of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,
9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 1.0 g, 1.29 mmol, 1.0 eq) in THF (10 ml) was added
picolinaldehyde (0.153 g, 1.42 mmol, 1.1 eq). The reaction mixture
was stirred at room temperature for about 30 minutes, then
sodiumtriacetoxyborohydride (0.60 g, 2.83 mmol, 2.2 eq) was added
and stirred at same temperature for overnight. TLC indicated
starting material was consumed and the desired product was
observed. The reaction mixture was cooled to 0.degree. C., pH
adjusted to 7.0 with 1N HCl, diluted with water (20 ml) and
extracted with CH.sub.2Cl.sub.2 (3.times.50 ml). The combined
organic extracts were dried over Na.sub.2SO.sub.4, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 5% MeOH in DCM as an
eluent to obtain the desired product (0.5 g, yield: 44.7%) as a
white solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 8.49
(d, J=4.2 Hz, 1H), 8.20 (brs, 1H), 7.75 (td, J=7.5, 1.5 Hz, 1H),
7.43-7.31 (m, 6H), 7.25 (dd, J=6.9, 5.1 Hz, 1H), 5.12, 5.05 (ABq,
J.sub.AB=12.6 Hz, 2H), 4.38-4.31 (m, 1H), 3.76-3.61 (m, 2H),
3.18-3.05 (m, 1H), 2.98-2.72 (m, 3H), 2.43-2.25 (m, 3H), 2.18 (d,
J=18.3 Hz, 1H), 2.0-1.72 (m, 5H), 1.70-1.42 (m, 5H), 1.40-1.20 (m,
13H), 1.18-1.0 (m, 9H), 0.90 (s, 3H), 0.89 (s, 3H), 0.86-0.77 (m,
13H); ESI-MS: m/z 862.7 (M+H).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,
8,11a-pentamethyl-3a-(2-methyl-2-((pyridin-2-ylmethyl)amino)propanamido)--
2-oxo-3,3a,4, 5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
[0558] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11a-pentamet-
hyl-3a-(2-methyl-2-((pyridin-2-ylmethyl)amino)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 1, 0.5 g,
0.58 mmol, 1.0 eq) in MeOH (10 ml), THF (10 ml) and water (1.6 ml)
was added KOH (0.227 g, 4.06 mmol, 7.0 eq). The reaction mixture
was stirred at room temperature for about 16 hours. TLC indicated
starting material was consumed and the desired product was
observed. The organic phase was evaporated under reduced pressure,
the reaction mixture was diluted with water (20 ml), cooled to
0.degree. C., pH adjusted to 7.0 with 1N HCl and extracted with DCM
(2.times.25 ml). The combined organic extracts were dried over
sodium sulfate, filtered and evaporated under reduced pressure. The
residue was purified by silicagel column chromatography by using 8%
MeOH in DCM as an eluent to obtain the desired product (350 mg,
yield: 78.1%) as a white solid. .sup.1H NMR (300 MHz,
DMSO-d.sub.6): .delta. ppm 8.49 (d, J=4.2 Hz, 1H), 8.20 (brs, 1H),
7.79-7.71 (m, 1H), 7.41 (d, J=7.5 Hz, 1H), 7.29-7.22 (m, 1H),
4.38-4.31 (m, 1H), 3.75-3.62 (m, 2H), 3.16-3.06 (m, 1H), 2.83-2.72
(m, 3H), 2.42-2.14 (m, 5H), 1.95-1.70 (m, 5H), 1.69-1.31 (m, 8H),
1.29-1.21 (m, 9H), 1.17-1.02 (m, 9H), 0.94-0.88 (m, 9H), 0.86-0.76
(m, 10H); ESI-MS: m/z 772.6 (M+H).sup.+; HPLC: 94.9%.
[0559] The below example 68 was prepared by the procedure similar
(including reagents and reaction conditions) to the above described
in the synthesis of example-67 using with their appropriate
intermediates.
Example 68
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((4-chlorobenzyl)a-
mino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3-
a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta-
[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00153##
[0561] Intermediate 1 was coupled with 4-chlorobenzaldehyde
followed by hydrolysis gave the desired product as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.62 (s, 1H), 7.25
(d, J=8.4 Hz, 2H), 7.17 (d, J=8.7 Hz, 2H), 4.38 (dd, J=11.1, 4.5
Hz, 1H), 4.17-4.12 (m, 1H), 3.65, 3.58 (ABq, J.sub.AB=13.5 Hz, 2H),
3.12-3.02 (m, 1H), 2.92-2.68 (m, 2H), 2.63-2.45 (m, 3H), 2.41-2.23
(m, 2H), 2.04-1.92 (m, 2H), 1.75-1.59 (m, 4H), 1.58-1.40 (m, 4H),
1.40-1.23 (m, 14H), 1.23-1.10 (m, 7H), 1.09-0.93 (m, 1H), 1.0 (s,
3H), 0.93-0.75 (m, 15H), 0.75-0.68 (m, 1H); ESI-MS: m/z 805.4
(M+H).sup.+; HPLC: 92.56%.
Example 69
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(((1-(4-chlorophen-
yl)cyclopropyl)methyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-o-
ctadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclob-
utane-1-carboxylic acid
##STR00154##
[0562] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(((1-(4-chloro
phenyl)cyclopropyl)methyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8-
,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00155##
[0564] To a stirred solution of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 1.0 g, 1.297 mmol, 1.0 eq) in 1,2-DCE (10 ml) at
0.degree. C. was added
1-(4-chlorophenyl)cyclopropane-1-carbaldehyde (Intermediate 18,
0.468 g, 2.595 mmol, 2.0 eq), followed by acetic acid (0.116 g,
1.945 mmol, 1.5 eq). The reaction mixture was stirred at 0.degree.
C. for about 30 minutes then sodiumtriacetoxy borohydride (1.09 g,
5.18 mmol, 4.0 eq) was added and allowed to stir at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The reaction mixture
was cooled to 0.degree. C., quenched with saturated ammonium
chloride solution (10 ml), diluted with water (30 ml) and extracted
with DCM (3.times.30 ml). The combined organic extracts were washed
with water (30 ml), dried over sodium sulfate, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-5% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the desired product (600 mg, yield: 49.4%) as a white solid.
.sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 7.37-7.23 (m, 9H),
5.15, 5.09 (ABq, J.sub.AB=12.3 Hz, 2H), 4.45 (dd, J=11.1, 4.5 Hz,
1H), 3.16-3.05 (m, 1H), 2.86-2.73 (m, 2H), 2.70-2.44 (m, 5H),
2.12-1.99 (m, 2H), 1.96-1.82 (m, 2H), 1.80-1.65 (m, 3H), 1.57-1.44
(m, 4H), 1.44-1.37 (m, 2H), 1.37-1.32 (m, 2H), 1.34 (s, 3H),
1.31-1.16 (m, 14H), 1.16-1.03 (m, 2H), 0.99 (s, 3H), 0.96 (s, 3H),
0.93-0.84 (m, 12H), 0.83-0.64 (m, 5H); ESI-MS: m/z 935.52
(M+H).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(((1-(4-chlorophen-
yl)cyclopropyl)methyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-2H-cyclopenta[a]
chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
[0565] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(((1-(4-chlorophenyl)cyclopr-
opyl)methyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,1-
3a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 1, 0.6 g,
0.641 mmol, 1.0 eq) in MeOH (10 ml) and THF (10 ml) was added
aqueous 2.5N KOH solution (1.78 ml, 4.49 mmol, 7.0 eq). The
reaction mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (30 ml),
cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (3.times.30 ml). The combined organic extracts
were washed with water (20 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-10% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the solid. To this solid compound, MTBE (5 ml) and hexane (5
ml) was added and heated to reflux for about 20 minutes. The
mixture was filtered and dried under vacuum to obtain the desired
product (90 mg, yield: 16.6%) as a white solid. .sup.1H NMR (300
MHz, CDCl.sub.3): .delta. ppm 7.36-7.20 (m, 4H), 4.47 (dd, J=11.1,
4.5 Hz, 1H), 3.18-3.05 (m, 1H), 2.88-2.76 (m, 2H), 2.68-2.60 (m,
1H), 2.60-2.52 (m, 2H), 2.50-2.44 (m, 2H), 2.13-2.0 (m, 2H),
1.92-1.83 (m, 2H), 1.77-1.63 (m, 3H), 1.63-1.41 (m, 6H), 1.38 (s,
3H), 1.37-1.29 (m, 3H), 1.26-1.10 (m, 14H), 1.08 (s, 3H), 1.05 (m,
1H), 1.0 (s, 3H), 0.93-0.84 (m, 12H), 0.83-0.73 (m, 4H), 0.72-0.65
(m, 1H); ESI-MS: m/z 845.44 (M+H).sup.+; HPLC: 91.5%.
Example 70
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(5-chloropicolinam-
ido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a-
,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[-
a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00156##
[0566] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(5-chloro
picolinamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl--
2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00157##
[0568] To a stirred solution of 5-chloropicolinic acid (0.286 g,
1.815 mmol, 2.0 eq) in DMF (7 ml) was added HATU (1.034 g, 2.721
mmol, 3.0 eq) followed by triethylamine (0.89 ml, 6.35 mmol, 7.0
eq). The reaction mixture was stirred at room temperature for about
30 minutes then
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropan
amido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,-
8,9,10,11,11a,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl) 3-benzyl
(1S,3R)-2,2-dimethyl cyclobutane-1,3-dicarboxylate (Intermediate 1,
0.700 g, 0.907 mmol, 1.0 eq) was added and heated at 60.degree. C.
for overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was diluted with
water (100 ml) and stirred at room temperature for about 30
minutes. The precipitates formed were collected by filtration,
washed with water (100 ml), and dried under vacuum to obtain the
solid. The resulting solid was purified by silicagel column
chromatography by using 0-2% methanol in dichloromethane gradient.
The fractions containing the expected product were combined and
concentrated under reduced pressure to obtain the desired product
(0.800 g, yield: 96.85%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 8.52 (d, J=2.1 Hz, 1H), 8.25 (brs, 1H),
8.13 (d, J=8.4 Hz, 1H), 7.85 (dd, J=8.4, 2.4 Hz, 1H), 7.56 (brs,
1H), 7.35 (m, 5H), 5.15, 5.09 (ABq, J.sub.AB=12.3 Hz, 2H), 4.44
(dd, J=11.1, 4.5 Hz, 1H), 3.22-3.10 (m, 1H), 2.87-2.57 (m, 5H),
2.40-2.22 (m, 2H), 2.09-2.0 (m, 1H), 1.98-1.80 (m, 3H), 1.78-1.70
(m, 2H), 1.68 (s, 3H), 1.65 (s, 3H), 1.53-1.40 (m, 4H), 1.37-1.33
(m, 4H), 1.31-1.0 (m, 12H), 0.96 (s, 3H), 0.93 (s, 3H), 0.92 (s,
3H), 0.87-0.76 (m, 10H).
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(5-chloro
picolinamido)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl--
2-oxo-3,3a,4,5,
5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
[0569] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-(5-chloropicolinamido)-2-met-
hylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 1, 0.800 g,
0.878 mmol, 1.0 eq) in MeOH (8 ml) and THF (8 ml) was added aqueous
2.5N KOH solution (2.63 ml, 6.585 mmol, 7.5 eq). The reaction
mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (10 ml),
cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (2.times.50 ml). The combined organic extracts
were washed with water (30 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-5% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the solid. To this solid compound, MTBE (10 ml) was added
and heated to reflux for about 30 minutes. The mixture was cooled
to 0.degree. C., solid was filtered and dried under vacuum to
obtain the desired product (0.048 g, yield: 6.6%) as an off-white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 8.52 (d,
J=2.1 Hz, 1H), 8.24 (brs, 1H), 8.13 (d, J=8.4 Hz, 1H), 7.85 (dd,
J=8.4, 2.1 Hz, 1H), 7.56 (s, 1H), 4.46 (dd, J=11.4, 4.8 Hz, 1H),
3.20-3.07 (m, 1H), 2.87-2.75 (m, 3H), 2.72-2.53 (m, 2H), 2.37-2.24
(m, 2H), 2.11-2.0 (m, 1H), 1.98-1.80 (m, 5H), 1.70-1.63 (m, 4H),
1.68 (s, 3H), 1.64 (s, 3H), 1.44-1.36 (m, 5H), 1.37 (s, 3H),
1.25-1.18 (m, 7H), 1.07 (m, 4H), 0.94 (s, 3H), 0.92 (s, 3H),
0.89-0.82 (m, 10H); ESI-MS: m/z 820.78 (M+H).sup.+; HPLC:
90.8%.
Example 71
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,811a-
-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-3-yl)ureido)
propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobuta-
ne-1-carboxylic acid
##STR00158##
[0570] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,75bR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-3-yl)ureido)propan-
amido)-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00159##
[0572] To a stirred solution of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 0.800 g, 1.037 mmol, 1.0 eq) in THF (20 ml) was
added DIPEA (0.73 ml, 4.148 mmol, 4.0 eq) and
5-isocyanato-2-methylpyridine (Intermediate 19, 0.139 g, 1.037
mmol, 1.0 eq). The reaction mixture was heated at 60.degree. C. for
about 16 hours. TLC indicated starting material was consumed and
the desired product was observed. The reaction mixture was diluted
with water (15 ml) and extracted with ethyl acetate (3.times.25
ml). The combined organic extracts were washed with water (25 ml),
dried over sodium sulfate, filtered and evaporated under reduced
pressure. The residue was purified by silicagel column
chromatography by using 3% methanol in dichloromethane as an eluent
to obtain the desired product (560 mg, yield: 59.7%) as a white
solid. .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. ppm 8.30 (d,
J=1.8 Hz, 1H), 7.94 (brs, 1H), 7.86 (dd, J=8.4, 2.1 Hz, 1H), 7.54
(s, 1H), 7.35 (m, 5H), 7.03 (d, J=8.4 Hz, 1H), 5.96 (s, 1H), 5.15,
5.09 (ABq, J.sub.AB=12.3 Hz, 2H), 4.43 (dd, J=10.8, 4.2 Hz, 1H),
3.20-3.08 (m, 1H), 2.98-2.90 (m, 1H), 2.88-2.72 (m, 2H), 2.70-2.58
(m, 2H), 2.49 (s, 3H), 2.35-2.20 (m, 2H), 2.20-1.83 (m, 8H),
1.63-1.40 (m, 2H), 1.49 (s, 3H), 1.47 (s, 3H), 1.40-1.10 (m, 12H),
1.34 (s, 3H), 1.10-0.75 (m, 20H); ESI MS: m/z 905.58
(M+H).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-3-yl)ureido)propan-
amido)-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)
carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
[0573] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-3-yl)-
ureido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 1, 0.700 g,
0.773 mmol, 1.0 eq) in MeOH (14 ml) and THF (14 ml) was added
aqueous 2.5 N KOH solution (2.32 ml, 5.80 mmol, 7.5 eq). The
reaction mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (20 ml),
cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (3.times.40 ml). The combined organic extracts
were washed with water (40 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-6% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
give a solid. To this solid compound, acetonitrile (15 ml) was
added and heated to reflux for about 30 minutes. The mixture was
cooled to 0.degree. C., filtered and dried under vacuum to obtain
the desired product (310 mg, yield: 49.1%) as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.1 (brs, 1H),
8.72 (s, 1H), 8.35 (d, J=2.4 Hz, 1H), 7.74 (dd, J=8.4, 2.4 Hz, 1H),
7.50 (s, 1H), 7.08 (d, J=8.7 Hz, 1H), 6.48 (s, 1H), 4.34 (m, 1H),
3.09 (m, 1H), 2.88-2.72 (m, 3H), 2.45-2.20 (m, 4H), 2.43 (s, 3H),
2.10 (d, J=17.7 Hz, 1H), 2.0-1.80 (m, 3H), 1.80-1.50 (m, 4H),
1.50-1.40 (m, 2H), 1.42 (s, 3H), 1.40 (s, 3H), 1.38-1.18 (m, 5H),
1.26 (s, 3H), 1.18-0.98 (m, 8H), 0.91 (s, 3H), 0.87 (s, 3H), 0.84
(s, 3H), 0.80 (s, 3H), 0.78 (s, 3H), 0.75-0.63 (m, 1H), 0.72 (s,
3H); ESI MS: m/z 815.45 (M+H).sup.+; HPLC: 95.0%.
Example 72
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-2-yl)ureido)
propanamido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octa-
decahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobuta-
ne-1-carboxylic acid
##STR00160##
[0574] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,
8,11a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-2-yl)ureido)propanam-
ido)-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl) (JR,
3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00161##
[0576] To a stirred solution of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 0.6 g, 0.778 mmol, 1.0 eq) in THF (6 ml) was added
triethylamine (0.54 ml, 3.89 mmol, 5.0 eq) and
2-isocyanato-6-methylpyridine (Intermediate 20, 0.208 g, 1.556
mmol, 2.0 eq). The reaction mixture was stirred at room temperature
for overnight. TLC indicated starting material was consumed and the
desired product was observed. The reaction mixture was evaporated
under reduced pressure, diluted with water (15 ml) and extracted
with DCM (2.times.25 ml). The combined organic extracts were washed
with water (25 ml), dried over sodium sulfate, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-5% MeOH in DCM gradient.
The fractions containing the expected product were combined and
concentrated under reduced pressure to obtain the desired product
(0.400 g, yield: 57.14%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 10.39 (s, 1H), 7.94 (s, 1H), 7.56-7.48 (m,
1H), 7.35 (m, 5H), 6.79 (d, J=7.5 Hz, 1H), 6.48 (d, J=8.1 Hz, 1H),
5.15, 5.09 (ABq, J.sub.AB=12.3 Hz, 2H), 4.44 (dd, J=11.4, 4.5 Hz,
1H), 3.20-3.08 (m, 1H), 2.90-2.58 (m, 5H), 2.46 (s, 3H), 2.35-2.23
(m, 2H), 2.09-1.98 (m, 1H), 1.97-1.86 (m, 3H), 1.80-1.67 (m, 2H),
1.64 (s, 3H), 1.61 (s, 3H), 1.55-1.10 (m, 17H), 1.34 (s, 3H), 1.08
(s, 3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.86-0.80 (m, 10H); ESI-MS:
m/z 905.76 (M+H).sup.+.
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,
8,8,11a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-2-yl)ureido)propan-
amido)-2-oxo-3, 3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,
11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,-
2-dimethylcyclobutane-1-carboxylic acid
[0577] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-iso
propyl-5a,5b,8,8,11a-pentamethyl-3a-(2-methyl-2-(3-(6-methylpyridin-2-yl)-
ureido)propan
amido)-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 1, 0.4 g,
0.441 mmol, 1.0 eq) in MeOH (4 ml) and THF (4 ml) was added aqueous
2.5N KOH solution (1.23 ml, 3.093 mmol, 7.0 eq). The reaction
mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (10 ml),
cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (2.times.30 ml). The combined organic extracts
were washed with water, dried over sodium sulfate, filtered and
evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-6% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
obtain the solid. To this solid compound, acetonitrile (20 ml) was
added and heated to reflux for about 30 minutes. The mixture was
cooled to 0.degree. C., filtered, was washed with acetonitrile (10
ml) and dried under vacuum to obtain the desired product (0.060 g,
yield: 16%) as an off-white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 10.5 (brs, 1H), 9.12 (s, 1H), 8.06 (s,
1H), 7.53 (t, J=7.8 Hz, 1H), 6.80 (d, J=7.2 Hz, 1H), 6.62 (d, J=7.8
Hz, 1H), 4.54-4.47 (m, 1H), 3.21-3.09 (m, 1H), 2.97-2.88 (m, 1H),
2.87-2.77 (m, 2H), 2.73-2.50 (m, 2H), 2.47 (s, 3H), 2.40-2.30 (m,
2H), 2.17-2.05 (m, 1H), 2.0-1.80 (m, 3H), 1.72-1.57 (m, 4H), 1.64
(s, 3H), 1.60 (s, 3H), 1.57-1.48 (m, 3H), 1.45-1.35 (m, 2H), 1.39
(s, 3H), 1.34-1.10 (m, 10H), 1.12 (s, 3H), 1.06 (s, 3H), 0.92 (s,
3H), 0.88 (s, 3H), 0.87-0.80 (m, 7H); ESI-MS: m/z 815.70
(M+H).sup.+; HPLC: 93.1%.
Example 73
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((2-(dimethylamino-
)ethyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-
-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cy-
clopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00162##
[0578] Step 1: Synthesis of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((2-(dimethyl
amino)ethyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentame-
thyl-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl) (JR,
3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
##STR00163##
[0580] To a stirred solution of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 1.0 g, 1.298 mmol, 1.0 eq) and sodium
2-(dimethylamino)-1-hydroxyethane-1-sulfonate (Intermediate 21,
1.241 g, 6.49 mmol, 5.0 eq) in methanol (10 ml) at 0.degree. C.
under nitrogen was added TEA (1 ml, 7.139 mmol, 5.5 eq). The
reaction mixture was stirred at 0.degree. C. for about 30 minutes
then sodium cyanoborohydride (0.163 g, 2.596 mmol, 2.0 eq) was
added and stirred at room temperature for overnight. TLC indicated
starting material was consumed and the desired product was
observed. The reaction mixture cooled to 0.degree. C., pH adjusted
to 7.0 with 1N HCl, and evaporated under reduced pressure. The
reaction mixture was diluted with DCM (75 ml), washed with water
(30 ml) and brine solution (20 ml). The combined organic extracts
were dried over Na.sub.2SO.sub.4, filtered and evaporated under
reduced pressure. The residue was purified by silicagel column
chromatography by using 0-10% methanol in dichloromethane gradient.
The fractions containing the expected product were combined and
concentrated under reduced pressure to obtain the desired product
(0.3 g, yield: 27.7%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 7.65 (brs, 1H), 7.38-7.33 (m, 5H), 5.15,
5.09 (ABq, J.sub.AB=12.3 Hz, 2H), 4.45 (dd, J=11.1, 4.5 Hz, 1H),
3.23-3.08 (m, 1H), 2.89-2.58 (m, 8H), 2.56-2.41 (m, 7H), 2.36-2.28
(m, 2H), 2.09-1.83 (m, 5H), 1.80-1.44 (m, 7H), 1.42-1.29 (m, 12H),
1.27-1.18 (m, 8H), 1.12 (s, 3H), 1.1-1.0 (m, 1H), 0.96 (s, 3H),
0.94 (s, 3H), 0.91 (s, 3H), 0.85 (s, 3H), 0.84 (s, 3H), 0.79 (m,
1H); ESI-MS: m/z 862.7 (M+H).sup.+ (100%).
Step 2: Synthesis of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((2-(dimethyl
amino)ethyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentame-
thyl-2-oxo-3,
3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopen-
ta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
[0581] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((2-(dimethylamino)ethyl)ami-
no)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-penta
methyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahy-
dro-2H-cyclo penta[a]chrysen-9-yl)
(1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate (step 1, 0.3 g,
0.356 mmol, 1.0 eq) in MeOH (5 ml) and THF (5 ml) was added aqueous
2.5N KOH solution (1.0 ml, 2.492 mmol, 7.0 eq). The reaction
mixture was stirred at room temperature for overnight. TLC
indicated starting material was consumed and the desired product
was observed. The organic phase was evaporated under reduced
pressure, the reaction mixture was diluted with water (15 ml),
cooled to 0.degree. C., pH adjusted to 5.0 with 1N HCl and
extracted with DCM (3.times.15 ml). The combined organic extracts
were washed with water (10 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-10% methanol in
dichloromethane gradient. The fractions containing the expected
product were combined and concentrated under reduced pressure to
give a solid. To this solid, acetonitrile (15 ml) was added and
heated to reflux for about 30 minutes. The mixture was filtered
through a Buchner funnel and was washed with acetonitrile (10 ml)
and dried under vacuum to obtain the desired product (60 mg, yield:
22.3%) as a white solid. .sup.1H NMR (300 MHz, Pyridine-d.sup.5):
.delta. ppm 8.43 (s, 1H), 4.82 (dd, J=11.1, 4.2 Hz, 1H), 3.44-3.33
(m, 1H), 3.22-3.04 (m, 5H), 2.80-2.68 (m, 2H), 2.66-2.57 (m, 2H),
2.57-2.47 (m, 3H), 2.29 (s, 6H), 2.22-1.90 (m, 5H), 1.90-1.70 (m,
3H), 1.65 (s, 3H), 1.61-1.50 (m, 18H), 1.48-1.39 (m, 5H), 1.30 (s,
3H), 1.27-1.18 (m, 1H), 1.09 (s, 3H), 1.06 (s, 3H), 1.03 (s, 3H),
0.98 (s, 3H), 0.94-0.88 (m, 1H); ESI-MS: m/z 752.66 (M+H).sup.+;
HPLC: 95.77%.
Example 74
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylprop-
anamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,
6,7,7a,8,9,10,1111a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-
-yl)oxy) carbonyl)-2,2-dimethylcyclobutane-1-carboxylic acid
hydrochloride
##STR00164##
[0583] To a stirred solution of
1-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-amino-2-methylpropanamido)-1-
-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,
10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)
3-benzyl (1S,3R)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1, 0.6 g, 0.778 mmol, 1.0 eq) in MeOH (10 ml) and THF
(10 ml) was added aqueous 2.5N KOH solution (2.18 ml, 5.44 mmol,
7.0 eq). The reaction mixture was stirred at room temperature for
overnight. TLC indicated starting material was consumed and the
desired product was observed. The organic phase was evaporated
under reduced pressure, the reaction mixture was diluted with water
(20 ml), cooled to 0.degree. C., pH adjusted to 5 with 1N HCl and
extracted with DCM (3.times.15 ml). The combined organic extracts
were washed with water (15 ml), dried over sodium sulfate, filtered
and evaporated under reduced pressure. The residue was purified by
silicagel column chromatography by using 0-10% methanol in
dichloromethane gradient, followed by recrystallization over
acetonitrile provided the solid. To this solid, 2N HCl in dioxane
(5 ml) was added and stirred at room temperature for overnight. The
mixture was evaporated under reduced pressure and recrystallization
over MTBE gave the desired product (0.44 g, yield: 80%) as a white
solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.16 (s,
1H), 8.17-8.08 (m, 4H), 4.40-4.32 (m, 1H), 3.16-3.06 (m, 1H),
2.84-2.67 (m, 3H), 2.32-2.22 (m, 4H), 2.02-1.83 (m, 3H), 1.80-1.55
(m, 5H), 1.53 (s, 3H), 1.49 (s, 3H), 1.47-1.32 (m, 6H), 1.27 (s,
3H), 1.31-1.23 (m, 1H), 1.22-1.10 (m, 9H), 1.06 (s, 3H), 0.92 (s,
3H), 0.91 (s, 3H), 0.87 (s, 3H), 0.83 (s, 3H), 0.82 (s, 3H);
ESI-MS: m/z 681.5 (M-HCl+H).sup.+; HPLC: 97.9%; chloride content by
Ion chromatography: 4.4%.
Example 75
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-butoxycarbo-
nyl)amino)-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-ox-
o-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclo-
penta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutane-1-carboxylic
acid
##STR00165##
[0585] To a stirred solution of 1-benzyl
3-((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((tert-butoxycarbonyl)amino)-
-2-methylpropanamido)-1-isopropyl-5a,5b,8,8,11a-pentamethyl-2-oxo-3,3a,4,5-
,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13,13a-octadecahydro-2H-cyclopenta[a]ch-
rysen-9-yl) (1R,3S)-2,2-dimethylcyclobutane-1,3-dicarboxylate
(Intermediate 1-step 12, 0.700 g, 0.803 mmol, 1.0 eq) in MeOH (7
ml) and THF (7 ml) was added aqueous 2.5N KOH solution (2.41 ml,
6.026 mmol, 7.5 eq). The reaction mixture was stirred at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The organic phase
was evaporated under reduced pressure, the reaction mixture was
diluted with water (10 ml), cooled to 0.degree. C., pH adjusted to
5.0 with 1N HCl and extracted with DCM (3.times.40 ml). The
combined organic extracts were washed with water (40 ml), dried
over sodium sulfate, filtered and evaporated under reduced
pressure. The residue was purified by silicagel column
chromatography by using 0-4% methanol in dichloromethane gradient.
The fractions containing the expected product were combined and
concentrated under reduced pressure to obtain the desired product
(0.245 g, yield: 39%) as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3): .delta. ppm 4.88 (s, 1H), 4.47 (dd, J=11.1, 4.8 Hz,
1H), 3.20-3.09 (m, 1H), 2.90-2.70 (m, 3H), 2.67-2.50 (m, 2H),
2.40-2.22 (m, 2H), 2.10-2.0 (m, 1H), 1.98-1.85 (m, 3H), 1.80-1.66
(m, 4H), 1.65-1.55 (m, 3H), 1.49 (s, 3H), 1.47 (s, 3H), 1.42 (s,
9H), 1.40-1.35 (m, 4H), 1.35-1.27 (m, 3H), 1.26-1.18 (m, 7H), 1.14
(s, 3H), 1.07 (s, 3H), 1.0-0.98 (m, 1H), 0.93 (s, 3H), 0.92 (s,
3H), 0.87 (s, 3H), 0.86 (s, 3H), 0.83-0.78 (m, 1H); ESI-MS: m/z
803.51 (M+Na).sup.+.
Example 76
Preparation of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-1-isopropyl-5a,5b,8,8,11-
a-pentamethyl-3a-(2-methyl-2-((S)-pyrrolidine-2-carboxamido)
propanamido)-2-oxo-3,3a,4,5,5a,5b,6,77a,8,9,10,11,11a,11b,12,13,13a-octad-
ecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethylcyclobutan-
e-1-carboxylic acid hydrochloride
##STR00166##
[0587] A solution of
(1R,3S)-3-((((3aR,5aR,5bR,7aR,9S,11aR,11bR,13aS)-3a-(2-((S)-1-(tert-butox-
ycarbonyl)pyrrolidine-2-carboxamido)-2-methylpropanamido)-1-isopropyl-5a,5-
b,8,8,
11a-pentamethyl-2-oxo-3,3a,4,5,5a,5b,6,7,7a,8,9,10,11,11a,11b,12,13-
,13a-octadecahydro-2H-cyclopenta[a]chrysen-9-yl)oxy)carbonyl)-2,2-dimethyl-
cyclobutane-1-carboxylic acid (Example 8, 0.400 g, 0.455 mmol, 1.0
eq) and 3N HCl in 1,4-dioxane (5 ml) was stirred at room
temperature for overnight. TLC indicated starting material was
consumed and the desired product was observed. The mixture was
evaporated under reduced pressure, the residue was washed with
n-hexane (10 ml), MTBE (10 ml) was added and heated to reflux for
about 30 minutes. The reaction mixture was cooled to 0.degree. C.,
filtered, solid was washed with MTBE (10 ml) and dried under vacuum
to obtain the desired product (0.340 g, yield: 91.64%) as a white
solid. .sup.1H NMR (300 MHz, DMSO-d.sub.6): .delta. ppm 12.16 (brs,
1H), 9.65 (brs, 1H), 8.52 (s, 1H), 8.46 (brs, 1H), 7.32 (s, 1H),
4.39-4.33 (m, 1H), 4.24-4.17 (m, 1H), 3.22-3.06 (m, 3H), 2.82-2.73
(m, 3H), 2.45-2.23 (m, 4H), 2.10 (d, J=13.8 Hz, 1H), 1.98-1.64 (m,
9H), 1.62-1.47 (m, 4H), 1.46-1.41 (m, 1H), 1.43 (s, 3H), 1.40-1.32
(m, 3H), 1.37 (s, 3H), 1.26 (s, 3H), 1.25-1.17 (m, 2H), 1.15-1.10
(m, 6H), 1.09-1.01 (m, 2H), 1.03 (s, 3H), 0.91 (s, 3H), 0.90 (s,
3H), 0.87 (s, 3H), 0.82 (s, 3H), 0.81 (s, 3H); ESI-MS: m/z 778.46
(M-HCl+H).sup.+; HPLC: 99.6%; chloride content by Ion
chromatography: 4.46%.
Biological Activity
[0588] The compounds described herein can be tested for their
antiviral activity following procedures known to a person of
ordinary skill in the art. For example, the following protocols can
be employed for testing the compounds. These protocols are
illustrative and do not limit to the scope of the invention.
Example 77
Evaluation of Compounds Antiviral Activity Against HIV-1 Strain
92HT599 in MT2 Cells
[0589] MT2 cells were infected with HIV-1 strain 92HT599 (10 TCID
50/30000 cells). The infected cells were plated at the
concentration of .about.30,000 cells per well in 96 well plate.
Test compound was added to the micro plate in defined format with
the final concentration of DMSO (vehicle) is not more than 1%.
Incubation was carried out in CO.sub.2 incubator for .about.96
hours for viral infection. At the end of incubation period an
aliquot from each well was taken for p24 estimation. The
quantitation of p24 is an index for antiviral activity of the
compound. Percent inhibition was calculated with reference to
control values (vehicle controls). p24 estimation was carried out
using Advance biosciences kit as per the procedure detailed by
supplier.
[0590] For 0% serum binding assay, wherein "A" refers to an
IC.sub.50 value of less than 3 nM, "B" refers to IC.sub.50 value in
range of 3.01-10 nM, and "C" refers to IC.sub.50 values greater
than 10 nM.
[0591] For 40% serum binding assay, wherein "A" refers to an
IC.sub.50 value of less than 50 nM, "B" refers to IC.sub.50 value
in range of 50.01-200 nM, and "C" refers to IC.sub.50 values
greater than 200 nM. The IC.sub.50 (nM) values are set forth in
Table-1.
TABLE-US-00001 TABLE 1 Example IC.sub.50 IC.sub.50 no 0% serum 40%
serum 1 A A 2 B A 3 A A 4 A A 5 A B 6 A B 7 A A 8 A A 9 A C 10 A A
11 A B 12 A B 13 A A 14 A A 15 A A 16 A B 17 A A 18 A A 19 A A 20 A
A 21 B A 22 A A 23 A A 24 A B 25 A A 26 A B 27 B B 28 A A 30 A A 31
A A 32 B C 33 A A 34 A A 35 C C 36 A B 37 A A 38 A A 39 A A 40 A A
41 A A 42 A A 43 A A 44 A B 45 B A 46 A A 47 A A 48 A A 49 A A 50 A
A 51 A A 53 A A 54 A A 55 B B 56 A A 57 A A 58 A A 59 A B 60 A A 61
A A 63 A A 64 A A 65 A A 67 A A 68 B A 69 C B 70 A A 71 A B 72 A A
73 A A 74 A A 75 A A 76 A B -- -- -- -- -- --
Example 78
Evaluation of Compounds Antiviral Activity Against pNL4-3/WT &
V7A Strains in MT4 Cells
[0592] MT4 cells were Transfected with HIV-1 Plasmid (pNL4-3-WT
& V7A) (Cells were incubating with required number of TCID50 of
HIV-1 for 1.5 h at 37.degree. C.). After infection, the infected
cells were plated at the concentration of 3.times.10.sup.4 cells
per well in 96 well plate. Test compound was added to the test
plate in defined format with the final concentration of DMSO is not
more than 1%. Incubation was carried out in CO.sub.2 incubator for
4 days for viral infection. At the end of incubation period an
aliquot from each well was taken for p24 estimation. p24 estimation
was carried out using Advance biosciences kit as per the procedure
detailed by supplier.
[0593] For pNL4-3 WT assay, IC.sub.50 wherein "A" refers to an
IC.sub.50 value of less than 5 nM, "B" refers to IC.sub.50 value in
range of 5.01-10 nM, and "C" refers to IC.sub.50 values greater
than 10 nM;
[0594] For pNL4-3 V7A assay, wherein "A" refers to an IC.sub.50
value of less than 10 nM, "B" refers to IC.sub.50 value in range of
10.01-50 nM, and "C" refers to IC.sub.50 values greater than 50 nM.
The IC.sub.50 (nM) values are set forth in Table-2.
TABLE-US-00002 TABLE 2 Example pNL4-3 WT pNL4-3 V7A no IC.sub.50
IC.sub.50 1 A A 5 A A 7 A A 14 A B 15 A C 16 B C 17 A A 18 A A 19 A
A 20 C C 25 A A 28 B B 29 A B 30 B C 31 A C 32 C C 33 A A 34 A B 37
A A 38 A A 39 B B 40 A A 41 C B 42 B A 44 C C 45 A B 47 A A 51 B C
53 A A 54 A B 55 C C 56 B B 57 B B 58 B C 61 A B 63 C C 64 B C 65 C
C 70 A A 72 A C
Example 79
Evaluation of Compounds Cyto-Toxicity (MTT Assay)
[0595] On day 1 calculate the number of cells required for the
assay and seed 3.times.104 cells in 200 .mu.l per well. Weigh the
compound and dissolve it in DMSO to get 10 mM stock which is
further diluted to 3 mM and 1 mM. The drugs from these stocks were
added to plate to get final concentration of 100 M, 30 M and 10 M.
Add DMSO to controls in a way to obtain final concentration of
solvent that is not greater than 1%. Incubate for 4 days in 5%
CO.sub.2 incubator at 37.degree. C. On day 4, 100 .mu.l of medium
was removed from each well without disturbing the cells. Add 10
.mu.l of MTT reagent and incubate for 4 hours at 5% CO.sub.2
incubator at 37.degree. C. for formation of crystals. Add 200 .mu.l
of 0.1N acidic isopropanol to dissolve the crystals and read the
plate at 590 nm. The values are mentioned in below Table-3 and
Table-3A.
TABLE-US-00003 TABLE 3 Cytotoxicity Example % viability No. 1 .mu.M
0.1 .mu.M 0.01 .mu.M 1 4 10 57 2 8 9 42 3 7 13 32 4 9 12 20 5 4 35
74 6 4 19 60 7 0 9 28 8 3 5 28 9 14 53 100 10 0 1 67 11 1 3 41 12 3
7 42 13 4 10 45 14 2 8 44 15 2 8 44 16 2 11 37 21 6 7 36 22 3 3 21
23 4 8 34 24 6 21 69 25 2 6 31 26 0 20 55 27 0 2 52 28 6 10 21 29 2
3 29 30 2 3 30 34 1 10 26 35 77 92 84 36 8 15 56 37 1 4 26 38 2 9
20 39 0 2 29 40 1 2 26 42 4 5 27 43 3 26 76 46 4 12 52 47 4 14 75
48 4 12 44 49 4 12 56 50 0 0 21 59 8 15 94 60 2 27 77 61 2 42 88 63
0 16 51 64 1 2 26 65 2 3 27 67 3 21 59 69 8 8 42 71 3 5 41 73 9 11
52 74 19 14 61 75 2 17 63 76 6 6 59 -- -- -- --
TABLE-US-00004 TABLE 3A Cytotoxicity Example % viability No. 0.01
.mu.M 0.003 .mu.M 0.001 .mu.M 17 26 62 81 18 29 61 78 19 40 65 79
20 40 85 91 31 54 86 81 44 58 80 84 51 37 85 86 52 59 89 83 72 27
84 98
Example 80
Evaluation of Compounds Single Dose Oral Pharmacokinetic Study
[0596] The test item was administered through oral route to animals
(rat/mice) at 30 mg/kg dose in a suitable vehicle (10% Solutol+20%
PEG) at 10 ml/kg dose volume. Blood samples (.about.50 .mu.L at
each time point) were collected from retro-orbital plexus using K3
EDTA as anticoagulant in eppendorf tubes at defined time intervals
30 minutes, 1 hour, 2 hour, 4 hour, 8 hour, 24 hour & 48 hour
under light ether anaesthesia. The samples were centrifuged at
3500.times.g to separate plasma and stored at -80.degree. C. until
analysis. Plasma 25 .mu.l for Mice were processed as per described
in sample preparation.
[0597] Standard solutions of the test compound 1 mg/mL solutions
were prepared in DMSO and further dilutions were made in methanol.
The calibration curve samples for LCMSMS analysis were prepared by
spiking 25 .mu.l of Mice plasma with 2.5 .mu.l and of the
appropriate working standard solution to obtain final
concentrations 0.078, 0.156, 0.312, 0.625, 1.25, 2.5, 5, 10, 20
& 40 .mu.g/ml. To the test compound plasma extraction was
carried out using Acetonitrile precipitation. After reconstitution
with solvent (50% Acetonitrile in Buffer) samples were analyzed by
LCMSMS to get the concentrations to calculate PK Parameters. The
values are set forth in Table-4.
TABLE-US-00005 TABLE 4 Mice oral PK @30 mg/kg Example C.sub.max
AUC.sub.0-t no (.mu.g/ml) (.mu.g hr/ml) 2 13.153 165.708 3 29.61
544.857 4 8.845 184.919 5 26.008 289.239 6 11.099 100.412 7 41.921
517.02 10 9.794 69.762 12 8.54 61.312 13 2.716 18.218 14 26.752
377.338 15 12.843 141.167 17 28.565 332.923 18 15.925 235.704 19
13.875 93.04 20 18.992 236.062 21 16.034 191.391 22 23.6 489.844 23
29.736 589.886 24 29.618 427.527 25 23.997 336.08 28 16.239 344.768
30 22.971 477.469 31 18.68 254.609 33 11.391 231.111 37 38.844
874.12 38 10.069 113.719 40 20.255 343.317 42 22.873 480.991 43
31.344 320.392 44 19.807 153.713 45 21.17 214.252 46 29.49 835.997
47 23.578 537.72 48 20.301 477.13 50 23.295 454.291 51 32.327
277.12 53 9.746 57.84 54 29.226 402.064 59 13.92 185.948 60 20.228
216.625 61 45.721 615.349 64 27.378 335.569 65 7.383 145.21 67
24.254 86.58 68 20.045 271.842 70 36.636 880.717 71 19.304 129.737
72 15.266 191.83
REFERENCES
[0598] 1. Antiviral methods and protocols (Eds: D Kinchington and
R. F. Schinazi) Humana Press Inc., 2000. [0599] 2. HIV protocols
(Eds: N. L. Michael and J. H. Kim) Humana Press Inc, 1999. [0600]
3. DAIDS Virology manual from HIV laboratories, Publication
NIH-97-3838, 1997. [0601] 4. HIV-1 p24 antigen capture assay,
enzyme immunoassay for detection of Human immunodeficiency Virus
Type 1 (HIV-1) p24 in tissue culture media--Advanced bio science
laboratories, Inc kit procedure.
[0602] Although the invention herein has been described with
reference to particular embodiments, it is to be understood that
these embodiments are merely illustrative of the principles and
applications of the present invention. It is therefore to be
understood that numerous modifications may be made to the
illustrative embodiments and that other arrangements may be devised
without departing from the spirit and scope of the present
invention as described above.
[0603] All publications and patent applications cited in this
application are herein incorporated by reference to the same extent
as if each individual publication or patent application was
specifically and individually indicated to be incorporated herein
by reference.
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