U.S. patent application number 09/900440 was filed with the patent office on 2002-06-13 for chemokine receptor antagonists.
This patent application is currently assigned to Warner-Lambert Company. Invention is credited to Bratton, Larry Don, Miller, Steven Robert, Roth, Bruce David, Trivedi, Bharat Kalidas, Unangst, Paul Charles.
Application Number | 20020072606 09/900440 |
Document ID | / |
Family ID | 26840401 |
Filed Date | 2002-06-13 |
United States Patent
Application |
20020072606 |
Kind Code |
A1 |
Bratton, Larry Don ; et
al. |
June 13, 2002 |
Chemokine receptor antagonists
Abstract
The instant invention is a series of compounds which are MCP-1
receptor antagonists of Formula I 1 Also included in the invention
are intermediates and processes for the preparation of the
compounds as well as methods of using the compounds as agents for
the treatment of atherosclerosis, chronic and acute inflammatory
disease, chronic and acute immune disorders and transplant
rejection as well as for preventing infection by HIV, treating
infection by TIV, delaying the onset of AIDS, or treating AIDS.
Inventors: |
Bratton, Larry Don;
(Whitmore Lake, MI) ; Miller, Steven Robert; (Ann
Arbor, MI) ; Roth, Bruce David; (Plymouth, MI)
; Trivedi, Bharat Kalidas; (Ann Arbor, MI) ;
Unangst, Paul Charles; (Ann Arbor, MI) |
Correspondence
Address: |
LAHIVE & COCKFIELD
28 STATE STREET
BOSTON
MA
02109
US
|
Assignee: |
Warner-Lambert Company
|
Family ID: |
26840401 |
Appl. No.: |
09/900440 |
Filed: |
July 5, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09900440 |
Jul 5, 2001 |
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09558267 |
Apr 25, 2000 |
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60142755 |
Jul 8, 1999 |
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Current U.S.
Class: |
546/50 |
Current CPC
Class: |
C07D 471/14
20130101 |
Class at
Publication: |
546/50 |
International
Class: |
C07D 471/14 |
Claims
1. A compound of Formula I 40or a pharmaceutically acceptable salt
thereof wherein: the --- means a double bond is possible; R.sub.1
is hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy, halogen,
CH.sub.2OH, trifluoromethyl, nitro, cyano, amino, substituted
amino, or (CH.sub.2).sub.m--COOR wherein m is an integer of from 0
to 2 and R is alkyl or hydrogen; R.sub.2 is hydrogen or lower
alkyl; R.sub.3 is lower alkyl, lower alkenyl, benzyl, or aryl;
R.sub.4 is hydrogen, lower alkyl, --COOR, --CONHR wherein R is
hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl, amino, or carbonyl;
and R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle with the proviso that R.sub.3 is not methyl when
R.sub.4 is hydrogen, R.sub.5 is hydroxy, and R.sub.6 is phenyl.
2. A compound according to claim 1 of formula 41wherein R.sub.1 is
hydrogen, bromine, or nitro; R.sub.3 is methyl, n-propyl, n-butyl,
or allyl; R.sub.4 is hydrogen or methyl; R.sub.5 is hydroxy, NOH,
or NH.sub.2; R.sub.7 is hydrogen, 3,4-methoxy, 3-methoxy,
3,5-methoxy, 3,4-chloro, 4-phenyl, or 4 42phenyl with the proviso
that R.sub.3 is not methyl when R4 and R.sub.7 are hydrogen, and
R.sub.5 is hydroxy
3. A method of inhibiting the binding of MCP-1 to its receptor
comprising administering a therapeutically effective amount of
compound according to claim 1 to a mammal in need thereof.
4. A compound according to claim 1 and selected from:
[2S-(2.alpha.,4a.alpha.,13.beta.,14.alpha..beta.)]-13b-Allyl-3-benzyliden-
e-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b-
]isoquinolin-2-ol;
[2S-(2.alpha.,4a.alpha.,13.beta.,14.alpha..beta.)]-3-Be-
nzylidine-13b-propyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',-
3':3,4]pyrido[1,2-b]-isoquinolin-2-ol;
[2S-(2.alpha.,4a.alpha.,13.beta.,14-
.alpha..beta.)]-3-Benzylidine-13b-butyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dod-
ecahydro-indolo[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-ol;
[2R-[2.alpha.(E),4a.alpha.,
13b.beta.,14a.beta.]]-3-Benzylidine-10-bromo--
13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyr-
ido[1,2-b]-isoquinolin-2-ol;
[2R-[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta-
.]]-3-Benzylidine-13b-methyl-10-nitro-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodec-
ahydro-indolo[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-ol;
[2R-[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]-3-Benzylidine-13b-methyl-
-12-nitro-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyri-
do[1,2-b]-isoquinolin-2-ol;
[2R-[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.-
]]-3-(3,4-Dimethoxy-benzylidene)-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-
-dodecahydro-indolo[2.alpha.,3.alpha.:3,4]pyrido[1,2-b]-isoquinolin-2-ol;
[2R-[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]-3-(3-Methoxy-benzylidene-
)-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]p-
yrido[1,2-b]-isoquinolin-2-ol; [2R-[2.alpha.(E),4a.alpha.,
13b.beta.,14a.beta.]]-3-(3,5-Dimethoxy-benzylidene)-13b-methyl-1,2,3,4,4a-
,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]-isoquinoli-
n-2-ol;
[2R-[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]-3-(3,4-Dichloro-b-
enzylidene)-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2-
',3':3,4]pyrido[1,2-b]-isoquinolin-2-ol;
[4aR-[(E),4a.alpha.,13b.beta.,14a-
.beta.]]-3-Benzylidene-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H--
indolo[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-one oxime;
3-Benzylidene-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indol-
o[2',3':3,4]pyrido[1,2-b]isoquinolin-2-ylamine;
[1R-[1.alpha.,2.beta.(E),4-
a.beta.,13b.alpha.,14a.beta.]]1,13b-Dimethyl-3-(4-styryl-benzylidene)-1,2,-
3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido]
1,2-b]isoquinolin-2-ol;
[1R-[1.alpha.,2.beta.(E),4a.beta.,13b.alpha.,14a.-
beta.]]3-Benzylidene-1,13b-dimethyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecah-
ydro-indolo[2',3':3,4]pyrido]1,2-b]isoquinolin-3-ol;
[1R-[1.alpha.,2.beta.(E),4a.beta.,13b.alpha.,14a.beta.]]3-Biphenyl-4-ylme-
thylene-1,13b-dimethyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2-
',3':3,4]pyrido] 1,2-b]isoquinolin-2-ol;
[4aS-(2E,4a.alpha.,13b.beta.,14a.-
beta.)]-13b-Allyl-3-benzylidene-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-in-
dolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-one;
[4aS-(2E,4a.alpha.,13.beta.-
,14a.beta.)]-3-Benzylidine-13b-propyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-
-1H-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-one;
[4aS-(2E,4a.alpha.,13.beta.,14a.beta.)]-3-Benzylidine-13b-butyl-3,4,4a,5,-
7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-
-one;
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-Benzylidine-10-bromo-13b-
-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,-
2-b]-isoquinolin-2-one;
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-Benzyl-
idine-13b-methyl-10-nitro-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2-
',3':3,4]pyrido[1,2-b]-isoquinolin-2-one and
[4aR-[(E),4a.alpha.,13b.beta.- ,
14a.beta.]]-3-Benzylidine-13b-methyl-12-nitro-3,4,4a,5,7,8,13,13b,14,14a-
-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-one;
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-(3,4-Dimethoxy-benzylidene)-1-
3b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[-
1,2-b]-isoquinolin-2-one;
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]3-(3-Me-
thoxy-benzylidene)-13b-methyl-3,4,4a,7,8,13,13b,14,14a-decahydro-1H-indolo-
[2',3':3,4]pyrido[1,2-b]isoquinolin-2-one;
[4aR-[(E),4a.alpha.,13b.beta.,1-
4a.beta.]]-3-(3,5-Dimethoxy-benzylidene)
13b-methyl-3,4,4a,5,7,8,13,13b,14-
,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-one;
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-(3,4-Dichloro-benzylidene)-13-
b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1-
,2-b]-isoquinolin-2-one; and
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-B-
enzylidene-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3'-
:3,4]pyrido[1,2-b]-isoquinolin-2-one.
5. A method of treating inflammation in a mammal in need thereof
comprising administering to said mammal an effective
anti-inflammatory amount of a compound of claim 1.
6. A method of treating inflammation in a mammal in need thereof
comprising administering to said mammal an effective
anti-inflammatory amount of a compound of claim 2.
7. A method of treating atherosclerosis in a mammal in need thereof
comprising administering to said mammal a therapeutically effective
amount of a compound of claim 1.
8. A method of treating pain in a mammal in need thereof comprising
administering to said mammal an effective analgesic amount of a
compound of claim 1.
9. A method of treating restenosis in a mammal in need thereof
comprising administering to said mammal a therapeutically effective
amount of a compound of claim 1.
10. A method of treating immune disorders in a mammal in need
thereof comprising administering to said mammal a therapeutically
effective amount of a compound of claim 1.
11. A method of treating transplant rejection in a mammal in need
thereof comprising administering to said mammal a therapeutically
effective amount of a compound of claim 1.
12. A method of treating a virus in a mammal in need thereof
comprising administering to said mammal an antivirally effective
amount of a compound of claim 1.
13. A method for preventing infection by HIV, treating infection by
HIV, delaying the onset of AIDS, or treating ADS comprising
administering to a mammal in need of said treatment a
therapeutically effective amount of a compound according to claim 1
or a pharmaceutically acceptable salt thereof.
14 A method for preventing infection by HIV, treating infection by
HIV, delaying the onset of AIDS, or treating AIDS comprising
administering to a mammal in need of said treatment a
therapeutically effective amount of a compound according to claim 2
or a pharmaceutically acceptable salt thereof.
15. A method for modulation of the CCR-5 chemokine receptor
activity in a mammal comprising administering an effective amount
of a compound according to claim 1 or a pharmaceutically acceptable
salt thereof.
16. A pharmaceutical composition comprising a compound according to
claim 1 in admixture with a pharmaceutically acceptable excipient,
diluent, or carrier.
17. A pharmaceutical composition comprising a compound according to
claim 2 in admixture with a pharmaceutically acceptable excipient,
diluent, or carrier.
18. A method of treating atherosclerosis in a mammal in need
thereof comprising administering a compound of Formula II 43or the
compound
[2R[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]3-Benzylidene-13b-methyl-1-
,2,3,4,4a,5,7,8,
13,13b,14,14adodecahydro-indolo[2',3':3,4]pyrido[1,2-b]is-
oquinolin-2-ol or a pharmaceutically acceptable salt thereof
wherein: the --- means a double bond is possible; R.sub.1 is
hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy, halogen,
CH.sub.2OH, trifluoromethyl, nitro, cyano, amino, substituted
amino, or (CH2).sub.m--COOR wherein m is an integer of from 0 to 2
and R is alkyl or hydrogen; R.sub.2 is hydrogen or lower alkyl;
R.sub.4 is hydrogen, lower alkyl, --COOR, --CONSR wherein R is
hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl, amino, or carbonyl;
and R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle.
19. A method of treating pain in a mammal in need thereof
comprising administering a compound of Formula II 44or the compound
[2R[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]3-Benzylidene-13b-methyl-1-
,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]is-
oquinolin-2-ol or a pharmaceutically acceptable salt thereof
wherein: the --- means a double bond is possible; R.sub.1 is
hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy, halogen,
CH.sub.2OH, trifluoromethyl, nitro, cyano, amino, substituted
amino, or (CH2).sub.m--COOR wherein m is an integer of from 0 to 2
and R is alkyl or hydrogen; R.sub.2 is hydrogen or lower alkyl;
R.sub.4 is hydrogen, lower alkyl, --COOR, --CONHR wherein R is
hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl, amino, or carbonyl;
and R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle.
20. A method of treating restenosis in a mammal in need thereof
comprising administering a compound of Formula II 45or the compound
[2R[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]3-Benzylidene-13b-methyl-1-
,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]is-
oquinolin-2-ol or a pharmaceutically acceptable salt thereof
wherein: the --- means a double bond is possible; R.sub.1 is
hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy, halogen,
CH.sub.2OH, trifluoromethyl, nitro, cyano, amino, substituted
amino, or (CH.sub.2).sub.m--COOR wherein m is an integer of from 0
to 2 and R is alkyl or hydrogen; R.sub.2 is hydrogen or lower
alkyl; R.sub.4 is hydrogen, lower alkyl, --COOR, --CONHR wherein R
is hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl, amino, or
carbonyl; and R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle.
21. A method of treating immune disorders in a mammal in need
thereof comprising administering a compound of Formula II 46or the
compound
[2R[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]3-Benzylidene-13b-methyl-1-
,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]is-
oquinolin-2-ol or a pharmaceutically acceptable salt thereof
wherein: the --- means a double bond is possible; R.sub.1 is
hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy, halogen,
CH.sub.2OH, trifluoromethyl, nitro, cyano, amino, substituted
amino, or (CH2).sub.m--COOR wherein m is an integer of from 0 to 2
and R is alkyl or hydrogen; R.sub.2 is hydrogen or lower alkyl;
R.sub.4 is hydrogen, lower alkyl, --COOR, --CONHR wherein R is
hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl, amino, or carbonyl;
and R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle.
22. A method of treating transplant rejection in a mammal in need
thereof comprising administering a compound of Formula II 47or the
compound
[2R[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]3-Benzylidene-13b-methyl-1-
,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]is-
oquinolin-2-ol or a pharmaceutically acceptable salt thereof
wherein: the --- means a double bond is possible; R.sub.1 is
hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy, halogen,
CH.sub.2OH, trifluoromethyl, nitro, cyano, amino, substituted
amino, or (CH.sub.2).sub.m--COOR wherein m is an integer of from 0
to 2 and R is alkyl or hydrogen; R.sub.2 is hydrogen or lower
alkyl; R.sub.4 is hydrogen, lower alkyl, --COOR, --CONER wherein R
is hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl, amino, or
carbonyl; and R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle.
23 A method of treating virus in a mammal in need thereof
comprising administering a compound of Formula II 48or the compound
[2R[2.alpha.(E),4a.alpha., 13b.beta.,
14a.beta.]]3-Benzylidene-13b-methyl-
-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]-
isoquinolin-2-ol or a pharmaceutically acceptable salt thereof
wherein: the --- means a double bond is possible; R.sub.1 is
hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy, halogen,
CH.sub.2OH, trifluoromethyl, nitro, cyano, amino, substituted
amino, or (CH.sub.2).sub.m--COOR wherein m is an integer of from 0
to 2 and R is alkyl or hydrogen,; R.sub.2 is hydrogen or lower
alkyl; R.sub.4 is hydrogen, lower alkyl, -COOR, -CONHR wherein R is
hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl, amino, or carbonyl;
and R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle.
24. A method for preventing infection by HIV, treating infection by
HIV, delaying the onset of AIDS, or treating AIDS in a mammal in
need thereof comprising administering a compound of Formula II 49or
the compound
[2R[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]3-Benzylidene-13b-methyl-1-
,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]is-
oquinolin-2-ol or a pharmaceutically acceptable salt thereof
wherein: the --- means a double bond is possible; R.sub.1 is
hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy, halogen,
CH.sub.2OH, trifluoromethyl, nitro, cyano, amino, substituted
amino, or (CH.sub.2).sub.m--COOR wherein m is an integer of from 0
to 2 and R is alkyl or hydrogen; R.sub.2 is hydrogen or lower
alkyl; R.sub.4 is hydrogen, lower alkyl, --COOR, --CONHR wherein R
is hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl, amino, or
carbonyl; and R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle.
25. A method for modulation of the CCR-5 chemokine receptor
activity in a mammal in need thereof comprising administering a
compound of Formula II 50or the compound
[2R[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]3-Benzyl-
idene-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3-
,4]pyrido[1,2-b]isoquinolin-2-ol or a pharmaceutically acceptable
salt thereof wherein: the --- means a double bond is possible;
R.sub.1 is hydrogen, lower alkyl, lower alkenyl, alkoxy, hydroxy,
halogen, CH.sub.2OH, trifluoromethyl, nitro, cyano, amino,
substituted amino, or (CH.sub.2).sub.m--COOR wherein m is an
integer of from 0 to 2 and R is alkyl or hydrogen; R.sub.2 is
hydrogen or lower alkyl; R.sub.4 is hydrogen, lower alkyl, --COOR,
--CONHR wherein R is hydrogen, alkyl, or aryl; R.sub.5 is hydroxyl,
amino, or carbonyl; and R.sub.6 is phenyl, substituted phenyl,
naphthyl, or a heterocycle.
Description
BACKGROUND OF TH INVENTION
[0001] U.S. Pat. No. 3,139,428 teaches compounds of formula 2
[0002] wherein R.sub.1 is hydrogen, methyl, or R.sub.3; R.sub.2 is
hydrogen, keto, hydroxyl or a radical of the formula --OR.sub.4 in
which R.sub.4 is the acyl radical of an aliphatic carboxylic acid
containing 2 to 6 carbon atoms such as acetyl, propionyl, butyryl,
valeryl, hexanoyl and the like, benzoyl or benzoyl substituted with
1, 2, or 3 lower alkyl, halo or lower alkoxy groups, for example
p-methylbenzoyl, m-methylbenzoyl, o-methylbenzoyl,
3,4-dimethylbenzoyl, p-chlorobenzoyl, p-fluorobenzoyl,
o-bromobenzoyl, m-chlorobenzoyl, 2,4-dichlorobenzoyl,
p-methoxybenzoyl, 3,4,5-trimethoxybenzoyl, 3,4-dimethoxybenzoyl and
the like; and R.sub.3 is --CHRs or --CH.sub.2R.sub.6 in which
R.sub.5 is lower alky, furyl, phenyl, phenyl lower alkyl such as
benzyl, phenethyl, phenylpropyl and the like, phenyl lower alkenyl
such as cinnamyl, phenylpropenyl, phenylbutenyl and the like,
methylenedioxyphenyl, or phenyl nuclearly substituted with 1 or 2
halo, lower alkyl, lower alkoxy or nitro groups such as
p-chlorophenyl, 3,4-dichlorophenyl, m-methylphenyl, o-bromophenyl,
p-ethylphenyl, p-fluorophenyl, p-methoxyphenyl, o-methoxyphenyl,
m-butoxyphenyl, p-nitrophenyl, 3,4-dinitrophenyl,
2,4-dinitrophenyl, o-nitrophenyl, m-nitrophenyl and the like, and
R.sup.6 is lower alkyl, furyl, phenyl, phenyl lower alkyl,
methylenedioxyphenyl, or phenyl nuclearly substituted with 1 or 2
halo, lower alkyl, lower alkoxy or amino groups. The compounds are
disclosed as useful as intermediates and as anti-inflammatory
agents.
[0003] U.S. Pat. No. 3,291,800 teaches compounds of formula 3
[0004] wherein R.sub.1 represents lower alkyl such as methyl,
ethyl, propyl, isopropyl, butyl, isobutyl and the like, aryl such
as phenyl or aralkyl such as benzyl; R.sub.2 represents hydrogen,
lower alkyl such as methyl, ethyl, propyl, isopropyl and the like,
lower alkoxy such as methoxy or ethoxy or acetyl; R.sub.3
represents hydrogen or lower alkyl and R.sub.4 represents hydrogen,
hydroxy or keto and to the nontoxic pharmaceutically acceptable
acid addition and quaternary ammonium salts thereof The compounds
are disclosed as having pharmacological activity as analgesics,
tranquilizers, and anti-inflammatory agents.
[0005] These patents are hereby incorporated by reference.
[0006] Migration of leukocytes from blood vessels into diseased
tissues is important to the initiation of normal disease-fighting
inflammatory responses. But this process, known as leukocyte
recruitment, is also involved in the onset and progression of
debilitating and life-threatening inflammatory and autoimmune
diseases. The pathology of these diseases results from the attack
of the body's immune system defenses on normal tissues. Thus,
blocking leukocyte recruitment to target tissues in inflammatory
and autoimmune disease would be a highly effective therapeutic
intervention. The leukocyte cell classes that participate in
cellular immune responses include lymphocytes, monocytes,
neutrophils, eosinophils and basophils. In many cases, lymphocytes
are the leukocyte class that initiates, coordinates, and maintains
chronic inflammatory responses, and thus are generally the most
important class of cells to block from entering inflammatory sites.
Lymphocytes attract monocytes to the site, which, collectively with
lymphocytes, are responsible for much of the actual tissue damage
that occurs in inflammatory disease. Infiltration of lymphocytes
and/or monocytes is responsible for a wide range of chronic,
autoimmune diseases, and also organ transplant rejection. These
diseases include, but are not limited to, rheumatoid arthritis,
atherosclerosis, psoriasis, chronic contact dermatitis,
inflammatory bowel disease, multiple sclerosis, sarcoidosis,
idiopathic pulmonary fibrosis, dermatomyositis, skin pemphigoid and
related diseases, (e.g., pemphigus vulgaris, p. foliacious, p.
erythematosis), glomerulonephritides, vasculitides, hepatitis,
diabetes, allograft rejection, and graft-versus-host disease.
[0007] This process, by which leukocytes leave the bloodstream and
accumulate at inflammatory sites, and initiate disease, takes place
in at least three distinct steps which have been described as (1)
rolling, (2) activation/firm adhesion and (3) transendothelial
migration (Springer T. A., Nature 1990;346:425-433; Lawrence and
Springer, Cell 1991;65:859-873; Butcher E. C., Cell
1991;67:1033-1036). The second step is mediated at a molecular
level by chemoattractant receptors. Chemoattractant receptors on
the surface of leukocytes bind chemoattractant cytokines secreted
by cells at the site of damage or infection. Receptor binding
activates leukocytes, increases the adhesiveness of the adhesion
molecules that mediate transendothelial migration, and promotes
directed migration of the cells toward the source of the
chemoattractant cytokine.
[0008] A recent discovery is the existence of a large family
(>20 members) of structurally homologous chemoattractant
cytokines, approximately 8 to 10 kD in size. These molecules share
the ability to stimulate directed cell migration (chemotaxis) and
have been collectively called "chemokines," a contraction of
chemotactic cytokines. Each chemokine contains four cysteine
residues (C) and two internal disulfide bonds. Chemokines can be
grouped into two subfamilies, based on whether the two amino
terminal cysteine residues are immediately adjacent (C--C family)
or separated by one amino acid (C--X--C family). These differences
correlate with the organization of the two subfamilies into
separate gene clusters. Within each gene cluster, the chemokines
typically show sequence similarities between 25% to 60%.
[0009] The chemokines of the C--X--C subfamily, such as
interleukin-8, are produced by a wide range of cell types and act
predominantly on neutrophils as mediators of acute inflammation.
Chemokines of the C--C subfamily are also produced by a wide
variety of cell types. These molecules act predominantly on subsets
of mononuclear inflammatory cells. Currently there are at least six
C--C chemokines with known chemotactic activity for human monocytes
and/or T cells, including MCP-1, MCP-2, MCP-3, MIP-1.alpha.,
MIP-1.beta., and RANTES. This suggests there may be a high degree
of redundancy in chemoattractant pathways. In addition, most C--C
chemokines are chemotactic for more than one cell type. For
examples, RANTES (regulated on activation, normal T cell expressed
and secreted) acts on memory CD4.sup.+ T cells, eosinophils, and
monocytes. Monocyte chemoattractant protein-I (MCP-1), another C--C
chemokine, acts on monocytes, activated "memory" T cells and on
basophils. MCP-1 is also a potent secretogogue of inflammatory
mediators for monocytes and basophils.
[0010] Five C--C chemokine receptors have recently been
characterized (CCRR1-5 or CCR1-CCR5), and all of these belong to
the seven transmembrane spanning G protein-coupled receptor family.
Each of these receptors mediates the binding and signaling of more
than one chemokine. For example, the CCR1 receptor binds both
MIP-1.alpha. and RANTES. There are 2 receptors which bind MCP-1,
CCR2 (with alternately spliced forms, 2A and 2B) and CCR4. CCR2 is
also known to mediate binding and signaling of MCP-3. The CCR4
receptor binds and signals, in addition to MCP-1, with RANTES and
MIP-1.alpha.. Which of these is responsible for the MCP-1 mediated
recruitment of monocytes and T cells is not known.
[0011] In agreement with the observation that lymphocyte emigration
into inflammatory sites is usually accompanied by emigration of
monocytes, MCP-1 is expressed at sites of antigen challenge and
autoimmune disease. However, analyses of human inflammatory lesions
with antibodies to other chemokines show RANTES, MIP-1I, MIP-1 and
MCP-3 to be present as well. Injection of MCP-1 into skin sites in
mice provokes only a mild monocytic infiltrate or no infiltrate at
all (Ernst C. A. et al., J. Immunol. 1994;152:3541-3544). Whether
these results reflect redundant and complex recruitment pathways
has not been resolved. MCP-1 and MCP-3 may play a role in allergic
hypersensitivity disease. This is suggested by the observation that
MCP-1 lacking the amino terminal glutamic acid loses the ability to
stimulate basophil mediator release and acquires activity as an
eosinophil chemoattractant.
[0012] Chemokines of both subfamilies may bind to heparan sulfate
proteoglycans on the endothelial cell surface, and may function
principally to stimulate haptotaxis of leukocytes that attach to
cytokine-activated endothelium through induced adhesion molecules.
Additionally, MCP-1 has been reported to selectively activate the
.beta.1 integrin family of leukocyte adhesion molecule, suggesting
a role in leukocyte interactions with the extracellular matrix.
Hence, MCP-1 may not only trigger the initial arrest and adhesion
of monocytes and T cells, but may also act to guide their migration
in extravascular space.
[0013] Chemoattractants appear to be required for transendothelial
migration in vitro and in vivo and can induce all steps required
for transmigration in vivo. Injection of neutrophil
chemoattractants into skin or muscle leads to robust emigration of
neutrophils from the vasculature and accumulation at the injection
site (Colditz, 1991). Pretreatment of neutrophils with pertussis
toxin inhibits emigration into inflammatory sites (Spangrude et
al., 1985; Nourshargh and Williams, 1990). Moreover, MAb to IL-8
markedly inhibits neutrophil emigration in inflammation (Sekido et
al., 1993).
[0014] Neutrophil chemoattractants injected into the same skin site
hours apart will stimulate neutrophil accumulation the first time
but not the second time, whereas a second injection into a distant
site will stimulate accumulation at that site. This desensitization
occurs for homologous chemoattractants only (Colditz, 1991) or
those that interact with the same receptor. Thus, chemoattractants
can act on and homologously desensitize a cell type that is
localized in tissue.
[0015] Chemokines mediate a range of proinflammatory effects on
leukocytes, such as chemotaxis, degranulation, and intigran
activation (Baggiolini et al., Adv. Immunol., 1994;55:97-179;
Oppenheim et al., Annu. Rev. Immunol., 1991; 9:61748; Miller et
al., Crit. Rev. Immunol., 1992;12:1746). These effects are mediated
by binding to the seven-transmembrane-spanning G-protein coupled
receptors (13aggiolini et al., Adv. Immunol., 1994;55:97-179;
Murphy, Annu. Rev. Immunol., 1994;12:593-633; Schall et al., Curr.
Opin. Immunol., 1994;6:865-73; Gerard et al., Curr. Opin. Immunol.,
1994;6;140-5; Mackay, Curr. Bio., In press). Chemokine receptors
also serve as coreceptors for HIV-1 entry into cells. This came
from observations that RANTES, MIP-1.alpha., and MIP-1.beta.
suppressed infection of susceptible cells in vitro by
macrophage-tropic primary HIV-1 isolates (Cocchi et al., Science
(Wash. D.C., 1995;270:1811-5). The chemokine receptor CXCR-4 was
found to support infection and cell fusion of CD4.sup.+ cells by
laboratory-adapted, T-tropic HIV-1 strains (Feng et al., Science
(Wash. D.C.), 1996;272:872-7). CCR-5, a RANTES, MIP-1.alpha., and
MIP-1.beta. receptor, was subsequently identified as the principle
coreceptor for primary macrophage-tropic strains (Choe et al.,
Cell, 1996;85: 1135-48; Alkhatib et al., Science (Wash. D.C.),
1996;272:1955-8, Doranz et al., Cell, 1996,85:1149-58, Deng et al.,
Nature (Lond.) 1996;381:661-6; Dragic et al., Nature (Lond),
1996;381:667-3). The importance of CCR-5 for HIV-1 transmission was
underscored by the observation that certain individuals who had
been repeatedly exposed to HIV-1 but remained uninfected had a
defect in CCR-5 expression (Liu et al., Cell, 1996; 86:367-77;
Samson et al., Nature (Lond), 1996;382:722-5; Dean et al., Science
(Wash. D.C.), 1996;273 :1856-62; Huang et al., Nature Med., 1996;2:
1240-3). These noninfectable individuals were found to be
homozygous for a defective CCR-5 allele that contains an internal
32-base pair deletion (CCR-5 .DELTA.32). The truncated protein
encoded by this gene is apparently not expressed at the cell
surface. CCR-5 .DELTA.32 homozygous individuals comprise .about.1%
of the Caucasian population and heterozygous individuals comprise
.about.20%. In studies of about 2700 HIV-1 infected individuals, no
.DELTA.32 homozygotes were found. Individuals who are heterozygous
for .DELTA.32 CCR-5 allele have been shown to progress more slowly
to AIDS than wild-type homozygous individuals (Samson et al.,
Nature (Lond.), 1996;382:722-5; Dean et al., Science (Wash. D.C.),
1996;273:1856-62; Huang et al., Nature Med., 1996;2:1240-3). Thus,
the identity of CCR-5 as the principle coreceptor for primary HIV
isolates provides an opportunity to understand disease
pathogenesis, and more importantly to identify a new avenue for the
treatment of HIV-1 infection.
[0016] Chemoattractants impart directionality to leukocyte
migration. By contrast with intradermal injection, intravascular
injection of IL-8 does not lead to emigration (Hechtman et al.,
1991). Cytokine-stimulated endothelial monolayers grown on filters
secrete IL8 into the underlying collagen layer. Neutrophils added
to the apical compartment emigrate into the basilar compartment,
but not when the IL-8 gradient is disrupted by addition of IL-8 to
the apical compartment (Huber et al., 1991).
[0017] The endothelium may present chemoattractants to leukocytes
in a functionally relevant way, as well as providing a permeability
barrier that stabilizes the chemoattractant gradient. Since
leukocytes, responding to specific antigen or inflammatory signals
in tissue, may signal emigration of further leukocytes into the
site, a chemoattractant was sought in material secreted by
mitogen-stimulated mononuclear cells (Carr et al. 1994)
Purification to homogeneity guided by a transendothelial lymphocyte
chemotaxis assay revealed that monocyte chemoattractant protein 1
(MCP-1), previously thought to be solely a monocyte
chemoattractant, is a major lymphocyte chemoattractant. An
activated subset of memory lymphocytes respond to MCP-1. In the
same assay, lymphocytes respond to RANTES and MIP-1.alpha. but less
so than to MCP-1 (C--C chemokines) and not at all to IL-8 or IP-10
(C--X--C chemokines). This physiologically relevant assay suggests
that C--C chemokines tend to attract both monocytes and
lymphocytes. In agreement with the observation that lymphocyte
emigration into inflammatory sites is accompanied by emigration of
monocytes, MCP-1 is abundantly expressed at sites of antigen
challenge and autoimmune disease (Miller and Krangel, 1992) and,
together with other chemokines, is an excellent candidate to
provide the step 2 signal required to activate integrin
adhesiveness and emigration of lymphocytes in vivo. (Traffic
Signals for Lymphocyte Recirculation and Leukocyte Emigration: The
Multistep Paradigm; Springer, Cell 1994;76: 301-314).
[0018] We have surprisingly found the compounds of the invention
are MCP-1 receptor antagonists and are capable of inhibiting the
binding of MCP-1 to its receptor. Surprisingly, the compounds block
T cell migration in vitro, and more surprisingly still, have
dramatic effects on the recruitment of inflammatory cells in
multiple models of inflammatory diseases. Thus, these compounds are
useful as agents for the treatment of inflammatory disease,
especially those associated with lymphocyte and/or monocyte
accumulation, such as arthritis, atherosclerosis and transplant
rejection. In addition, these compounds can be used in the
treatment of allergic hypersensitivity disorders such as asthma and
allergic rhinitis characterized by basophil activation and
eosinophil recruitment, as well as for the treatment of restenosis
and chronic or acute immune disorders.
[0019] The importance of CCR-5 for HIV-1 transmission was
underscored by the observation that certain individuals who had
been repeatedly exposed to HIV-1 but remained uninfected had a
defect in CCR-5 expression (Liu et al., Cell, 1996; 86:367-77;
Samson et al., Nature (Lond.), 1996;382:722-5; Dean et al., Science
(Wash. D.C.), 1996;273:1856-62; Huang et al., Nature Med.,
1996;2:1240-3). These noninfectable individuals were found to be
homozygous for a defective CCR-5 allele that contains an internal
32-base pair deletion (CCR-5 .DELTA.32). The truncated protein
encoded by this gene is apparently not expressed at the cell
surface. CCR-5 .DELTA.32 homozygous individuals comprise .about.1%
of the Caucasian population and heterozygous individuals comprise
.about.20%. In studies of about 2700 HIV-1 infected individuals, no
.DELTA.32 homozygotes were found. Individuals who are heterozygous
for .DELTA.32 CCR-5 allele have been shown to progress more slowly
to AIDS than wild-type homozygous individuals (Samson et al.,
Nature (Lond.), 1996;382:722-5; Dean et al., Science (Wash. D.C.),
1996;273:1856-62; Huang et al., Nature Med., 1996;2: 1240-3). Thus,
the identity of CCR-5 as the principle coreceptor for primary HIV
isolates provides an opportunity to understand disease
pathogenesis, and more importantly to identify a new avenue for the
treatment of HIV-1 infection.
SUMMARY OF THE INVENTION
[0020] The invention is a novel compound of Formula I: 4
[0021] or a pharmaceutically acceptable salt thereof wherein
R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, and R.sub.6 are as
described below.
[0022] The instant invention is also a method of inhibiting the
binding of MCP-1 to its receptor comprising administering a
therapeutically effective amount of a compound of Formula I above.
The compounds are also useful for modulation of the CCR-5 chemokine
receptor activity.
[0023] The compounds of Formula II 5
[0024] are useful in treating atherosclerosis, pain, restenosis,
immune disorders, preventing transplant rejection, treating virus,
HV, AIDS and for modulation of the CCR-5 chemokine receptor
activity.
[0025] The compounds of Formula I are useful in the treatment of
atherosclerosis, chronic and acute inflammatory disease, chronic
and acute immune disorders, and transplant rejection in mammals in
need of such treatment. The compounds are also useful for
preventing infection by HIV, treating infection by HIV, delaying
the onset of AIDS, or treating AIDS.
[0026] Novel intermediates useful in the preparation of the final
compounds and novel synthetic methods are also part of the instant
invention.
[0027] Pharmaceutical compositions of one or more of the compounds
of Formula I use a further part of this invention.
DETAILED DESCRIPTION OF THE INVENTION
[0028] The instant invention is a compound of Formula I 6
[0029] or a pharmaceutically acceptable salt thereof wherein:
[0030] the--means a double bond is possible;
[0031] R.sub.1 is hydrogen, lower alkyl, lower alkenyl, alkoxy,
hydroxy, halogen, CH.sub.2OH, trifluoromethyl, nitro, cyano, amino,
substituted amino, or (CH.sub.2).sub.m--COOR wherein m is an
integer of from 0 to 2 and R is alkyl or hydrogen;
[0032] R.sub.2 is hydrogen or lower alkyl;
[0033] R.sub.3 is lower alkyl, lower alkenyl, benzyl, or aryl;
[0034] R.sub.4 is hydrogen, lower alkyl, --COOR, --CONSR wherein R
is hydrogen, alkyl, or aryl;
[0035] R.sub.5 is hydroxyl, amino, or carbonyl; and
[0036] R.sub.6 is phenyl, substituted phenyl, naphthyl, or a
heterocycle with the proviso that R.sub.3 is not methyl when
R.sub.4 is hydrogen, R.sub.5 is hydroxy, and R.sub.6 is phenyl.
[0037] The invention is also methods of treatment comprising
administering a compound of Formula II or the compound
[2R[2.alpha.(E),4a.alpha.,13b.be-
ta.,14a.beta.]]3-Benzylidene-13b-methyl-1,2,3,4,4a,5,7,8,
13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-ol.
These methods are: atherosclerosis, restenosis, immune disorders,
transplant rejection, virus, preventing infection by HIV, treating
infection by HIV, delaying the onset of AIDS, and treating AIDS.
The compounds are also useful for modulating the CCR-5 receptor
activity.
[0038] In the compounds of Formula I and II, the term lower alkyl
means a straight or branched alkyl of from 1 to 4 carbon atoms and
includes, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl,
sec-butyl, isobutyl, or tert-butyl.
[0039] The lower alkenyl means a straight or branched carbon chain
of from 2 to 6 carbons with one or more double bonds.
[0040] The alkoxy is o-alkyl as defined above for alkyl.
[0041] Halogen is fluorine, chlorine, bromine, or iodine.
[0042] Amino is optionally substituted by one or more selected from
methyl, ethyl, n-propyl, or i-propyl.
[0043] Phenyl may optionally be substituted with from 1 to 3
substituents selected from lower alkyl, alkoxy, hydroxy, halogen,
trifluoriomethyl, nitro, cyano, amino, substituted amino,
COOR.sub.7, CONR.sub.8R.sub.9, or phenyl wherein R.sub.7, R.sub.8,
and R.sub.9 are each independently selected from hydrogen, alkyl,
or aryl.
[0044] Preferably, heterocycle is 5- or 6-membered mono- or
bicyclic ring structures which may contain one or more heteroatom
such as N or O; examples of heterocycle are pyrrole, furan,
thiophene, benzofuran, benzothiophene, pyridine, pyrimidine,
pyridazine, pyrazole, oxazole, indole, N-alkylindole, quinoline,
quinazoline, quinazolinone and the like. Substituents can be
hydrogen, alkyl of from 1 to 4 carbon atoms; cycloalkyl of from 5
to 7 carbon atoms, alkoxy, hydroxy, --CN, halogen, trifluoromethyl,
nitro, amino, or substituted amino.
[0045] Aryl is unsubstituted or substituted phenyl or naphthyl.
[0046] The term "mammal" includes animals and humans.
[0047] Some of the compounds of Formula I and Formula II are
capable of further forming both pharmaceutically acceptable acid
addition and/or base salts. All of these forms are within the scope
of the present invention.
[0048] Pharmaceutically acceptable acid addition salts of the
compounds of Formula I include salts derived from nontoxic
inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric,
hydrobromic, hydriodic, hydrofluoric, phosphorous, and the like, as
well as the salts derived from nontoxic organic acids, such as
aliphatic mono- and dicarboxylic acids, 2-phenyl-substituted
alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic
acids, aliphatic and aromatic sulfonic acids, etc. Such salts thus
include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite,
nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate,
metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate,
trifluoroacetate, propionate, caprylate, isobutyrate, oxalate,
malonate, succinate, suberate, sebacate, fumarate, maleate,
mandelate, benzoate, chlorobenzoate, methylbenzoate,
dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate,
phenylacetate, citrate, lactate, maleate, tartrate,
methanesulfonate, and the like. Also contemplated are salts of
amino acids such as arginate and the like and gluconate,
galacturonate (see, for example, Berge S. M. et al.,
"Pharmaceutical Salts," J. Pharma Sci., 1977;66:1).
[0049] The acid addition salts of said basic compounds can be
prepared by contacting the free base form with a sufficient amount
of the desired acid to produce the salt in the conventional manner.
The free base form may be regenerated by contacting the salt form
with a base and isolating the free base in the conventional manner.
The free base forms can differ from their respective salt forms
somewhat in certain physical properties such as solubility in polar
solvents.
[0050] Pharmaceutically acceptable base addition salts can be
formed with metals or amines, such as alkali and alkaline earth
metals or organic amines. Examples of such metals used as cations
are sodium, potassium, magnesium, calcium, and the like. Examples
of suitable amines are N,N'-dibenzylethylenediamine,
chloroprocaine, choline, diethanolamine, dicyclohexylamine,
ethylenediamine, N-methylglucamine, and procaine (see Berge, Supra,
1977).
[0051] The base addition salts of said acidic compounds can be
prepared by contacting the free acid form with a sufficient amount
of the desired base to produce the salt in the conventional manner.
The free acid form may be regenerated by contacting the salt form
with an acid and isolating the free acid in the conventional
manner. The free acid forms can differ from their respective salt
forms somewhat in certain physical properties such as solubility in
polar solvents.
[0052] Certain of the compounds of the present invention can exist
in unsolvated forms as well as solvated forms, including hydrated
forms and are intended to be encompassed within the scope of the
present invention.
[0053] Certain of the compounds of the present invention possess
one or more chiral centers and each center may exist in the R(D) or
S(L) configuration. The present invention includes all enantiomeric
and epimeric forms as well as the appropriate mixtures thereof The
compounds are valuable agents for the treatment of inflammatory
diseases or conditions, atherosclerosis, restenosis, and autoimmune
disorders such as arthritis and transplant rejection.
[0054] In a preferred embodiment, the disease or condition is one
which is associated with lymphocyte and/or monocyte infiltration of
tissues (including recruitment and/or accumulation in tissues),
such as arthritis (e.g., rheumatoid arthritis), inflammatory bowel
diseases (e.g., Crohn's disease, ulcerative colitis), multiple
sclerosis, idiopathic pulmonary fibrosis, and graft rejection
(e.g., in transplantation), including allograft rejection or
graft-versus-host disease. In addition, diseases characterized by
basophil activation and/or eosinophil recruitment, including
allergic hypersensitivity disorders such as asthma and allergic
rhinitis can be treated according to the present invention.
[0055] Other diseases that may be treated with the compounds of
Formula I are: psoriasis, chronic contact dennatitis, sarcoidosis,
dermatomyositis, skin phemphigoid and related diseases (e.g.,
pemphigus vulgaris, p. foliacious, p. erythematosus),
glomerulonephritides, vasculitides (e.g., necrotizing, cutaneous,
and hypersensitivity vasculitis), hepatitis, diabetes, systemic
lupus erythematosus and myasthenia gravis.
[0056] In addition to psoriasis, other inflammatory dermatoses such
as dermatitis, eczema, atopic dermatitis, allergic contact
dermatitis, urticaria and reperfusion injury can also be
treated.
[0057] MCP-1 Binding Assay
[0058] Membranes used in the MCP-1 binding assay were prepared from
TBP-1 cells (human monocytic cell line source--American Type
Culture Collection, Tumor Immunology Bank #202, Rockville, Md.,
accession No. ATCC TIB 202). Cells were harvested by
centrifiugation and washed twice in ice-cold PBS
(phosphate-buffered saline) and the cell pellet was frozen at
-80.degree. C. in some cases. Cells were resuspended in ice-cold
lysis buffer 5 mM HEPES (2-(4N-[2-hydroxyethyl]piperazin-1-yl)-N-
'-(2-ethanesulfonic acid), pH 7.5, 2 mM EDTA
(ethylenediaminetetraacetic acid), 5 .mu.g/mL each leupeptin,
aprotinin, chymostatin (protease inhibitors), and 100 .mu.g/mL PMSF
(phenylmethane sulfonyl fluoride--also a protease inhibitor)) at a
concentration of 5.times.10.sup.7 cells/mL. The cell suspension was
dounced 10 to 15 times using the B pestle (small pestle of tissue
grinder--clearance is 0.07 mm; source--Fisher Scientific) on ice.
Nuclei and debris were removed by centrifugation at 500 to
1000.times. g for 10 minutes at 4.degree. C. The supernatant was
transferred to a fresh tube and centrifuged at 25,000.times. g for
30 minutes at 4.degree. C. The supernatant was aspirated, and the
pellet was resuspended in freezing buffer (10 mM HEPES, pH 7.5, 300
mM sucrose, 1 .mu.g/mL each leupeptin, aprotinin, chymostatin, and
10 .mu.g/mL PMSF) using a mini-homogenizer until all clumps were
resolved. Membranes were aliquoted and frozen at minus 70.degree.
C. to 85.degree. C. until needed. Typical binding assays used 10 to
20 .mu.g/well of total membrane protein as determined with a
standard protein assay (e.g. Bradford protein assay, BioRad,
Richmond, Calif.).
[0059] For binding, 10 to 20 .mu.g of total membrane protein were
included in the binding reaction along with 0.2 nM
I.sup.125-labeled MCP-1 (Amersham, Arlington Heights, Ill.) with or
without unlabeled competitor MCP-1 (Peprotech, Rocky Hill, N.J.)
(at 500 nM). Binding reactions were performed in a final volume of
100 .mu.L in a binding buffer containing 10 mM HEPES, pH 7.2, 1 mM
CaCl.sub.2, 5 mM MgCl.sub.2, 0.5% BSA (bovine serum albumin). After
30 to 60 minutes at room temperature, the binding reactions were
filtered through GF/C filters (Whatman glass fiber filters, Type C)
or GF/B unifilter plates (Packard) which had been pre-soaked with
0.3% polyethyleneimine and washed twice with binding buffer
containing 0.5 M NaCl. Filters were dried and counted in a
Beta-Plate scintillation counter using standard scintillation
fluid. Final concentration of compound in the binding assay ranged
from 0.05 to 100 TM. Compounds were dissolved in DMSO (dimethyl
sulfoxide). Final concentrations of DMSO in the binding were kept
constant at 0.5%.
[0060] IC.sub.50s were calculated using a non-linear 3-parameter
logistic curve fit. IC.sub.50 means the concentration at which 50%
inhibition is achieved. Negative controls contained the same amount
of DMSO vehicle as used in wells containing compound. Positive
control contained 250 to 500 nM cold competitor MCP-1 in DMSO
vehicle. Non-specific binding (the level of bound .sup.125I-labeled
MCP-1 in the presence of 250 to 500 TM unlabeled MCP-1) was
subtracted from all data prior to analysis.
[0061] The compounds of the invention have IC.sub.50s between about
1.0 to 50 micromolar at CCR-2 receptor.
[0062] CCR-5 Receptor Binding Assay
[0063] The .sup.125I-gp120/sCD4/CCR-5 binding assay was carried out
similarly as described in Wu et al., Nature, 1996;384:179-183.
Briefly, the envelope gp120 protein derived from HIV-1 JR-FL
(Trkola et al., Nature, 1996;384:184-186), a M-tropic strain, was
iodinated using solid phase lactoperoxidase to a specific activity
of 20 .mu.Ci/.mu.g. For each binding reaction (in a final volume of
100 .mu.L binding buffer [50 mM HBEPES, pH 7.5, 1 mM CaCl.sub.2, 5
mM MgCl.sub.2, and 0.5% BSA]), 25 .mu.L (2.5 .mu.g) of membranes
prepared from CCR-5/L 1.2 cells were mixed with 25 .mu.L (3 nM)
sCD4, followed by 25 .mu.L (0.1 nM) radio-labeled gp 120 in the
presence or absence of 25 .mu.L compound dissolved in DMSO (final
concentration of DMSO 0.5%). The reactions were incubated at room
temperature for 45 to 60 minutes and stopped by transferring the
mixture to GFB filter plates, which were then washed 3 to 4 times
with binding buffer containing 0.5 M NaCl. The plates were dried
and MicroScint scintillation fluid was added before counting.
[0064] The compounds of the invention have IC.sub.50s between about
0.2 to 50 micromolar at the CCR-5 receptor.
[0065] The compounds of the present invention can be prepared and
administered in a wide variety of routes of administration such as
parenteral, oral, topical, rectal, inhalation and the like.
Formulations will vary according to the route of administration
selected. Examples are oral and parenteral dosage forms. Thus, the
compounds of the present invention can be administered by
injection, that is, intravenously, intramuscularly,
intra-cutaneously, subcutaneously, intraduodenally, or
intra-peritoneally. Also, the compounds of the present invention
can be administered by inhalation, for example, intranasally.
Additionally, the compounds of the present invention can be
administered transdermally. The following dosage forms may comprise
as the active component, a compound of Formula I or a corresponding
pharmaceutically acceptable salt of a compound of Formula I.
[0066] For preparing pharmaceutical compositions from the compounds
of the present invention, pharmaceutically acceptable carriers can
be either solid or liquid. Solid form preparations include powders,
tablets, pills, capsules, cachets, suppositories, and dispersible
granules. A solid carrier can be one or more substances which may
also act as diluents, flavoring agents, binders, preservatives,
tablet disintegrating agents, or an encapsulating material.
[0067] In powders, the carrier can be a finely divided solid which
is in a mixture with the finely divided active component.
[0068] In tablets, the active component can be mixed with the
carrier having the necessary binding properties in suitable
proportions and compacted in the shape and size desired.
[0069] The powders and tablets preferably contain from 5% or 10% to
about 70% of the active compound. Suitable carriers are magnesium
carbonate, magnesium stearate, talc, sugar, lactose, pectin,
dextrin, starch, gelatin, tragacanth, methylcellulose, sodium
carboxymethylcellulose, a low melting wax, cocoa butter, and the
like The term "preparation" is intended to include the formulation
of the active compound with encapsulating material as a carrier
providing a capsule in which the active component with or without
other carriers, is surrounded by a carrier, which is thus in
association with it. Similarly, cachets and lozenges are included.
Tablets, powders, capsules, pills, cachets, and lozenges can be
used as solid dosage forms suitable for oral administration.
[0070] For preparing suppositories a low melting wax, such as a
mixture of fatty acid glycerides or cocoa butter, is first melted
and the active component can be dispersed homogeneously therein, as
by stirring. The molten homogenous mixture can be then poured into
convenient sized molds, allowed to cool, and thereby to
solidify.
[0071] Liquid form preparations include solutions, suspensions, and
emulsions, for example, water or water propylene glycol solutions.
For parenteral injection, liquid preparations can be formulated in
solution in a pharmaceutically acceptable carrier, such as, aqueous
polyethylene glycol solution.
[0072] Aqueous solutions suitable for oral use can be prepared by
dissolving the active component in water and adding suitable
colorants, flavors, stabilizing, and thickening agents as
desired.
[0073] Aqueous suspensions suitable for oral use can be made by
dispersing the finely divided active component in water or another
suitable carrier with viscous material, such as natural or
synthetic gums, resins, methylcellulose, sodium
carboxymethylcellulose, and other well-known suspending agents.
[0074] Also included are solid form preparations which are intended
to be converted, shortly before use, to liquid form preparations
for oral administration. Such liquid forms include solutions,
suspensions, and emulsions. These preparations may contain, in
addition to the active component, colorants, flavors, stabilizers,
buffers, artificial and natural sweeteners, dispersants,
thickeners, solubilizing agents, and the like.
[0075] The pharmaceutical preparation is preferably in unit dosage
form. In such form the preparation is subdivided into unit doses
containing appropriate quantities of the active component. The unit
dosage form can be a packaged preparation, the package containing
discrete quantities of preparation, such as packeted tablets,
capsules, and powders in vials or ampoules. Also, the unit dosage
form can be a capsule, tablet, cachet, or lozenge itself, or it can
be the appropriate number of any of these in packaged form.
[0076] The quantity of active component in a unit dose preparation
may be varied or adjusted for example from about 0.1 mg to 200 mg,
preferably about 0.5 mg to 100 mg according to the particular
application and the potency of the active component. The
composition can, if desired, also contain other compatible
therapeutic agents.
[0077] In therapeutic use as agents for the treatment of
inflammatory diseases, inflammatory diseases, atherosclerosis,
restenosis, and immune disorders such as arthritis and transplant
rejection, the compounds utilized in the pharmaceutical methods of
this invention can be administered at an initial dosage of about
0.01 mg to about 200 mg/kg daily. A daily dose range of about 0.01
mg to about 50 mg/kg is preferred. The dosages, however, may be
varied depending upon the requirements of the patient, the severity
of the condition being treated, and the compound being employed.
Determination of the proper dosage for a particular situation is
within the skill of the art. Generally, treatment is initiated with
smaller dosages which are less than the optimum dose of the
compound. Thereafter, the dosage is increased by small increments
until the optimum effect under the circumstances is reached. For
convenience, the total daily dosage may be divided and administered
in portions during the day, if desired.
[0078] General Preparative Methods for Compounds of the
Invention
[0079] In the IUPAC systematic nomenclature, these compounds are
named as derivatives of indolo[2',3':3,4]pyrido[1,2-b]isoquinoline.
In this system, the 13b-, 2-, and 3-substituent positions
correspond to the 3-, 17-, and 18-positions, respectively, in the
older nomenclature based on the natural product yohimbine.
[0080] Imminium salts of
indolo[2',3':3,4]pyrido[1,2-b]isoquinolines 1 are reacted with an
alkyl or aryl lithium reagent or an alkyl or aryl Grignard reagent
to yield the 13b-alkyl or aryl derivatives 2 (Scheme 1 below). The
reaction may be carried out in tetrahydrofuiran or diethyl ether at
0 to 70 degrees for 1 to 24 hours, following the method of Zinnes
H., Comes R. A., and Shavel J., Jr., J. Org. Chem., 1965;30:105.
The starting imminium salts 1 are prepared as described in the
above reference, or by Godtfredsen W. O. and Vangedal S., Acta
Chem. Scand, 1956;10:1414. Compounds of type 2 in which R.sub.3 is
hydrogen are also prepared from the natural product yohimbine as
described by Aube J. and Ghosh S., Advances in Natural Product
Synthesis, 1996;3:99, and Baxter E. W. and Mariano P. S., in
Alkaloids: Chemical and Biological Perspectives; Pelletier S. W.,
Ed., Springer-Verlag: New York, 1992;8: 197-319. Alternatively, the
imminium salts 1 are reacted with allyltributyl tin or tetraallyl
tin to produce the 13b-allyl derivatives 3. The reaction may be run
in an alcohol such as methanol, ethanol, or 2-methoxyethanol, or a
mixed solvent system consisting of an alcohol and chloroform or
dichloromethane at 40 to 125 degrees for 1 to 24 hours. The
13b-allyl adducts 3 thus prepared may be subjected to catalytic
hydrogenation to obtain the corresponding 13b-propyl analogs 4.
[0081] The compounds of type 2 are oxidized at the 2-carbinol
substituent to yield the ketone derivatives 5 (Scheme 2 below). The
oxidation may be performed by the Oppenauer method utilizing an
aluminum alkoxide reagent (Zinnes H., Comes R. A., and Shavel J.,
Jr., reference cited above), or with tetrapropylammonium
perruthenate (TPAP; Ley S. V., Norman J., Griffith W. P., and
Marsden S. P., Synthesis, 1994:639), or with dimethyl sulfoxide and
oxalyl chloride (Swem oxidation; Tidwell T. T., Organic Reactions,
1990;39:297). Condensation of the ketones 5 with an aromatic
aldehyde 6 in methanol or ethanol at 50 to 80 degrees for 2 to 24
hours in the presence of aqueous sodium hydroxide or potassium
hydroxide yields the benzylidene ketones 7 when R.sub.6 is phenyl
or substituted phenyl. The benzylidene moiety may be reduced to a
benzyl substituent by standard catalytic hydrogenation methods.
Reduction of the ketones 7 with sodium borohydride or potassium
borohydride in methanol, ethanol, or tetrahydrofuran at 25 to 80
degrees for 2 to 24 hours gives the carbinols 8 (Shavel J., Jr.,
Bobowski G., and von Strandtmann M., U.S. Pat. No. 3,139,428).
[0082] The benzene ring of the indole portion of these compounds
may be subjected to standard aromatic substitution reactions, such
as bromination and nitration, in order to prepare compounds 10-15
(Schemes 3 and 4). In addition, the ketones 7 may be converted to
oximes 16 and the oximes reduced to obtain amines 17 (Scheme 5). 7
8 9 10 11
[0083] The present invention is further directed to combinations of
the present compounds with one or more agents useful in the
prevention or treatment of AIDS. For example, the compounds of this
invention may be effectively administered, whether at periods of
pre-exposure and/or post-exposure, in combination with effective
amounts of the anti-HIV compounds, immunomodulators,
anti-infectives, or prophactic or therapeutic vaccines known to
those of ordinary skill in the art.
1 ANTIVIRALS Drug Name Manufacturer Indication 097 Hoechst/Bayer
HIV infection, AIDS, ARC (non-nucleoside reverse transcriptase (RT)
inhibitor) GW141 W94/ Glaxo Wellcome HIV infection, AIDS, ARC VX478
(protease inhibitor) Amprenavir GW1592U89 Glaxo Wellcome HIV
infection, AIDS, ARC Abacavir (RT inhibitor) Acemannan Carrington
Labs ARC (Irving, TX) Acyclovir Burroughs Wellcome HIV infection,
AIDS, ARC, in Combination with AZT AD-439 Tanox Biosystems HIV
infection, AIDS, ARC AD-519 Tanox Biosystems HIV infection, AIDS,
ARC Adefovir dipivoxil Gilead Sciences HIV infection AL-721 Ethigen
ARC, PGL HIV positive, (Los Angeles, CA) AIDS Alpha Interferon
Glaxo Wellcome Kaposi's sarcoma, HIV in combination Alferon
Interferon Interferon Sciences Kaposi's sarcoma, HIV in combination
Ansamycin Adria Laboratories ARC LM 427 (Dublin, OH) Erbamont
(Stamford, CT) Antibody which Advanced Biotherapy AIDS, ARC
neutralizes pH Concepts labile alpha (Rockville, MD) aberrant
Interferon AR177 Aronex Pharm HIV infections, AIDS, ARC
beta-fluoro-ddA Nat'l Cancer Institute AIDS-associated diseases
BMS-232623 Bristol-Myers HIV infection, AIDS, ARC (CGP-73547)
Squibb/Novartis (protease inhibitor) BMS-234475 Bristol-Myers HIV
infection, AIDS, ARC (CGP-61755) Squibb/Novartis (protease
inhibitor) (-)6-Chloro-4(S)- Merck HIV infection, AIDS,
cyclopropylethynyl- ARC (non-nucleoside 4(S)-trifluoro- reverse
transcriptase methyl-1,4-dihydro- inhibitor) 2H-3,1 -benzoxazin-
2-one CI-1012 Warner-Lambert HIV-1 infection Cidofovir Gilead
Science CMV retinitis, herpes, papillomavirus Combivir AZT+3TC
Glaxo Wellcome HIV infection, AIDS, ARC Curdlan sulfate AJI Pharma
USA HIV infection Cytomegalovirus MedImmune CMV retinitis immune
globin Cytovene Ganciclovir Syntex/Roche Sight threatening CMV,
peripheral CMV, retinitis Delaviridine Pharmacia-Upjohn HIV
infection, AIDS, ARC (RT inhibitor) Dextran Sulfate Ueno Fine Chem.
Ind. AIDS, ARC, HIV Ltd. (Osaka, Japan) positive asymptomatic HIVID
(ddc) Hoffman-La Roche HIV infection, AIDS, Dideoxycytidine ARC ddI
Dideoxyinosine Bristol-Myers Squibb HIV infection, AIDS, ARC;
combination with AZT/d4T DMP-450 Triangle HIV infection, AIDS,
Pharmaceutical ARC (protease inhibitor) Efavirenz (DMP 266) DuPont
Merck HIV infection, AIDS, ARC (non-nucleoside RT inhibitor) EL10
Elan Corp, PLC HIV infection (Gainesville, GA) Famciclovir Smith
Kline Herpes zoster, herpes simplex Foscavir/Foscarnet Astra CMV,
HSV 1-2 FTC Triangle HIV infection, AIDS, Pharmaceutical ARC
(reverse transcriptase inhibitor) GS 840 Gilead HIV infection,
AIDS, ARC (reverse transcriptase inhibitor) HBY097 Hoechst Marion
HIV infection, AIDS, Roussel ARC (non-nucleoside reverse
transcriptase inhibitor) Hypericin VIMRx Pharm. HIV infection,
AIDS, ARC Recombinant Human Triton Biosciences AIDS, Kaposi's
sarcoma, Interferon Beta (Almeda, CA) ARC Interferon alpha-n3
Interferon Sciences ARC, AIDS Indinavir Merck HIV infection, AIDS,
ARC, asymptomatic HIV positive, also in combination with AZT/
ddI/ddC ISIS 2922 ISIS Pharmaceuticals CMV retinitis JE 2147
(KNI-764) Japan Energy/ HIV infection, AIDS, ARC Protease inhibitor
Agouron PI (reverse transcriptase inhibitor); also with AZT KNI-272
Nat'l Cancer Institute HIV-associated diseases Lamivudine, 3TC
Glaxo Wellcome HIV infection, AIDS, ARC (reverse transcriptase
inhibitor); also with AZT Lobucavir Bristol-Myers Squibb CMV
infection - HBV infection Nelfinavir Agouron HIV infection, AIDS,
Pharmaceuticals ARC (protease inhibitor) Nevirapine Boeheringer HIV
infection, AIDS, Ingleheim ARC (RT inhibitor) Novapren Novaferon
Labs, Inc. HIV inhibitor (Akron, OH) Peptide T Peninsula Labs AIDS
Octapeptide (Belmont, CA) Sequence PNU-140690 Pharmacia Upjohn HIV
infection, AIDS, ARC (protease inhibitor) Probucol Vyrex HIV
infection, AIDS RBD-CD4 Sheffield Med. Tech HIV infection, AIDS,
(Houston, TX) ARC Ritonavir Abbott HIV infection, AIDS, ARC
(protease inhibitor) S-1153 Agouron/Shionogi NnRTI Saquinavir
Hoffmann-La Roche HIV infection, AIDS, ARC (protease inhibitor)
Stavudine; d4T Bristol-Myers Squibb HIV infection, AIDS,
Didehydrodeoxy- ARC thymidine Valaciclovir Glaxo Wellcome Genital
HSV & CMV infections Virazole Ribavirin Viratek/ICN
Asymptomatic HIV (Costa Mesa, CA) positive, LAS, ARC Zidovudine;
AZT Glaxo Wellcome HIV infection, AIDS, ARC, Kaposi's sarcoma, in
combination with other therapies
[0084]
2 IMMUNO-MODULATORS Drug Name Manufacturer Indication AS-101
Wyeth-Ayerst AIDS Bropirimine Pharmacia Upjohn Advanced AIDS
Acemannan Carrington Labs, Inc. AIDS, ARC (Irving, TX) CL246, 738
American Cyanamid AIDS, Kaposi's sarcoma Lederle Labs EL10 Elan
Corp, PLC HIV infection (Gainesville, GA) FP-21399 Fuki ImmunoPharm
Blocks HIV fusion with CD4+ cells Gamma Interferon Genentech ARC,
in combination w/ TNF (tumor necrosis factor) Granulocyte Genetics
Institute AIDS Macrophage Sandoz Colony Stimulating Factor
Granulocyte Hoeschst-Roussel AIDS Macrophage Immunex Colony
Stimulating Factor Granulocyte Schering-Plough AIDS, combination w/
Macrophage AZT Colony Stimulating Factor HIV core Particle Rorer
Seropositive HIV Immunostimulant IL-2 Interleukin-2 Cetus AIDS, in
combination w/ AZT IL-2 Interleukin-2 Hoffman-La roche AIDS, ARC,
HIV, in Immunex combination w/AZT IL-2 Interleukin-2 Chiron AIDS,
increase in CD4 (aldeslukin) cell counts Immune Globulin Cutter
Biological Pediatric AIDS, in Intravenous (Berkeley, CA)
combination w/AZT (human) IMREG-1 Imreg AIDS, Kaposi's sarcoma,
(New Orleans, LA) ARC, PGL IMREG-2 Imreg AIDS, Kaposi's sarcoma,
(New Orleans, LA) ARC, PGL Imuthiol Diethyl Merieux Institute AIDS,
ARC Dithio Carbamate Alpha-2 Interferon Schering Plough Kaposi's
sarcoma w/ AZT, AIDS Methionine- TNI Pharmaceutical AIDS, ARC
Enkephalin (Chicago, IL) MTP-PE Muramyl- Ciba-Geigy Corp. Kaposi's
sarcoma Tripeptide Granulocyte Colony Amgen AIDS, in combination w/
Stimulating Factor AZT Remune Immune Response Immunotherapeutic
Corp. rCD4 Recombinant Genentech AIDS, ARC Soluble Human CD4
rCD4-IgG hybrids AIDS, ARC Recombinant Soluble Biogen AIDS, ARC
Human CD4 Interferon Alfa 2a Hoffman-La Roche Kaposi's sarcoma
AIDS, ARC, in combination w/ AZT SK&F106528 Smith Kline HIV
infection Soluble T4 Thymopentin Immunobiology HIV infection
Research Institute (Annandale, NJ) Tumor Necrosis Genentech ARC, in
combination w/ Factor; TNF gamma Interferon
[0085]
3 ANTI-INFECTIVES Drug Name Manufacturer Indication Clindamycen
with Pharmacia Upjohn PCP Primaquine Fluconazole Pfizer
Cryptococcal meningitis, candidiasis Pastille Nystatin Squibb Corp.
Prevention of oral Pastille candidiasis Ornidyl Eflornithine
Merrell Dow PCP Pentamidine LyphoMed PCP treatment Isethionate (IM
& (Rosemont, IL) IV) Trimethoprim Antibacterial
Trimethoprim/sulfa Antibacterial Piritrexim Burroughs Wellcome PCP
treatment Pentamidine Fisons Corporation PCP prophylaxis
isethionate for inhalation Spiramycin Rhone-Poulenc Cryptosporidial
diarrhea Intraconazole- Janssen Pharm. Histoplasmosis; R51211
cryptococcal meningitis Trimetrexate Warner-Lambert PCP
[0086]
4 OTHER Drug Name Manufacturer Indication Daunorubicin NeXstar,
Sequus Karposi's sarcoma Recombinant Human Ortho Pharm. Corp.
Severe anemia associated Erythropoietin with AZT therapy
Recombinant Human Serono AIDS-related wasting, Growth Hormone
cachexia Megestrol Acetate Bristol-Myers Squibb Treatment of
anorexia associated w/AIDS Testosterone Alza, Smith Kline
AIDS-related wasting Total Enteral Norwich Eaton Diarrhea and
Nutrition Pharmaceuticals malabsorption related to AIDS
[0087] It will be understood that the scope of combinations of the
compounds of this invention with AIDS antivirals, immunomodulators,
anti-infectives or vaccines is not limited to the list in the above
table, but includes in principle any combination with any
pharmaceutical composition useful for the treatment of AIDS.
[0088] The following examples are illustrative of the instant
invention. They do not limit the scope of the invention.
EXAMPLE 1
[0089] 12
[0090]
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)]-13b-Butyl-1,2,3,4,4a,5-
,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-
-ol
[0091] A suspension of
[2R-(2.alpha.,4a.alpha.,14.alpha..beta.)]-2-hydroxy-
-2,3,4,4a,5,7,8,13,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoqu-
inolin-6-ylium chloride (10.6 g, 32.0 mmol; Zinnes H., Comes R. A.,
and Shavel J., Jr., J. Org. Chem., 1965;30:105) in 600 mL of
diethyl ether was cooled in an ice bath while a solution of n-butyl
lithium (100 mL, 160 mmol of 1.6 M in hexane) was added dropwise.
The mixture was stirred at room temperature for 18 hours, then at
reflux for 6 hours. The mixture was again cooled in ice and treated
dropwise with 200 mL of saturated aqueous ammonium chloride
solution. The mixture was filtered, and the filtrate layers were
separated. The aqueous layer was extracted with ethyl acetate, and
the extracts were combined with the original organic layer. The
insoluble material was digested several times with warm ethyl
acetate. The mixture was filtered, and the filtrate was added to
the original ethyl acetate extracts. The combined organic layers
were washed with brine, dried (Na.sub.2SO.sub.4), and evaporated.
The residue was purified by flash chromatography (2% triethylamine
in ethyl acetate) to yield 1.8 g (16%) of product. A sample
recrystallized twice from aqueous 2-propanol had mp 235.degree. C.
dec.; .sup.1H NMR (DMSO-d.sub.6).delta. 0.63-0.79 (m, 4H),
0.82-1.15 (m, 2H), 1.17-1.28 (m, 5H), 1.35-1.52 (m, 3H), 1.61-1.72
(m, 3H), 1.87 (m, 1H), 2.13 (m, 1H), 2.55-2.76 (m, 4H), 2.87 (m,
1H), 3.09 (m, 1H), 3.46 (m, 1H), 4.49 (d, J=4.6 Hz, 1H), 6.89-7.00,
(m, 2H), 7.24 (d, J=8.0 Hz, 1H), 7.32 (d, J=7.5 Hz, 1H), 10.59 (s,
1H); MS (APCI.sup.+), m/z 353 (MH.sup.+).
EXAMPLE 2
[0092] 13
[0093] [2R-(2.alpha.,4a.alpha.,
13.beta.,14a.beta.)]-13b-Allyl-1,2,3,4,4a,-
5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin--
2-ol
[0094] A suspension of [2R-(2.alpha.,4a.alpha.,
14a.beta.)]-2-hydroxy-2,3,-
4,4a,5,7,8,13,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoquinoli-
n-6-ylium chloride (11.2 g, 33.9 mmol) and tetraallyl tin (10.5 mL,
12.4 g, 43.7 mmol) in 100 mL of 2-methoxyethanol was stirred at
reflux for 4 hours. The cooled reaction mixture was added to 1.0 L
of 5% aqueous sodium bicarbonate solution and 1.0 L of ethyl
acetate. The mixture was filtered, and the filtrate layers were
separated. The aqueous layer was extracted with fresh ethyl
acetate, and the extracts were combined with the original organic
layer. The original insoluble material was digested several times
with warm ethyl acetate. The mixture was filtered, and the filtrate
was added to the original ethyl acetate extracts. The combined
organic layers were washed several times with brine, dried
(Na.sub.2SO.sub.4), and evaporated. The residue was purified by
flash chromatography (0.75% triethylamine in ethyl acetate) to
yield 6.6 g (58% of product. A sample recrystallized from aqueous
2-propanol had mp 228-230.degree. C.; .sup.1H NMR
(DMSO-d.sub.6).delta. 0.87-1.27 (m, 4H), 1.35-1.51 (m, 3H), 1.68
(m, 1H), 1.78-1.93 (m, 2H), 2.52-2.73 (m, 5H), 2.81-3.07 (m, 3H),
3.47 (m, 1H), 4.51 (d, J=4.3 Hz, 1H), 4.81 (dd, J=2.2, 8.2 Hz, 1H),
4.92 (dd, J=2.2, 15.2 Hz, 1H), 5.52 (m, 1H), 6.89-7.01 (m, 2H),
7.25 (d, J=8.0 Hz, 1H), 7.33 (d, J=7.7 Hz, 1H), 10.6 (s, 1H); MS
(APCI.sup.+), m/z 337 (MH.sup.+).
EXAMPLE 3
[0095] 14
[0096]
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)]-13b-Propyl-1,2,3,4,4a,-
5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin--
2-ol
[0097] A solution of
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)]-13b-ally-
l-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b-
]-isoquinolin-2-ol (5.3 g, 15.8 mmol) in 100 mL of 50%
tetrahydrofuran/methanol was hydrogenated over 0.5 g of 10%
palladium on carbon catalyst. The catalyst was filtered, and the
filtrate was evaporated to yield 5.1 g (96%) of the product. A
sample purified by flash chromatography (10% methanol in
dichloromethane) and recrystallized from ethyl acetate/hexane had
mp 195.degree. C. dec.; .sup.1H NMR (DMSO-d.sub.6).delta. 0.75-0.76
(m, 4H), 0.83-1.06 (m, 2H), 1.08-1.32 (m, 3H), 1.33-1.52 (m, 3H),
1.58-1.77 (m, 3H), 1.87 (m, 1H), 2.07 (m, 1H), 2.53-2.75 (m, 4H),
2.88 (m, 1H), 3.09 (m, 1H), 3.45 (m, 1H), 4.49 (d, J=4.6 Hz, 1H),
6.89-7.00 (m, 2H), 7.24 (d, J=8.0 Hz, 1H), 7.32 (d, J=7.7 Hz, 1H),
10.59 (s, 1H); MS (APCI.sup.+), m/z 339 (MH.sup.+).
EXAMPLE 4
[0098] 15
[0099]
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)]-13b-Butyl-3,4,4a,5,7,8-
,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-blisoquinolin-2-on-
e
[0100] A solution of oxalyl chloride (0.52 mL, 0.76 g, 6.0 mmol) in
15 mL of dichloromethane was cooled to -78.degree. C. while a
solution of dimethyl sulfoxide (1.0 mL, 1.1 g, 14.1 mmol) in 5.0 mL
of dichloromethane was added dropwise. The mixture was stirred for
15 minutes, and a solution of
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)]-1-
3b-butyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrid-
o[1,2-b]isoquinolin-2-ol (1.8 g, 5.1 mmol) in 25 mL of
tetrahydrofuran was added dropwise. The mixture was stirred for 30
minutes, and N,N'-diisopropylethylamine (4.5 mL, 3.3 g, 25.8 mmol)
was added dropwise. The cooling bath was removed, and the mixture
was stirred for 16 hours. The reaction mixture was added to 400 g
of ice and water and extracted with ethyl acetate. The combined
organic layers were washed with brine, dried (Na2SO.sub.4), and
evaporated. The residue was purified by flash chromatography
(50:50:1 ethyl acetate/hexane/triethylamine) to yield 1.4 g (76%)
of product of mp 235.degree. C. dec.; .sup.1H NMR
(DMSO-d.sub.6).delta. 0.63-0.77 (m, 4H), 1.09-1.41 (m, 4H),
1.55-1.87 (m, 6H), 2.01-2.29 (m, 4H), 2.35-2.47 (m, 1H), 2.57-2.72
(m, 2H), 2.73-2.82 (m, 2H), 2.90-2.97 (m, 1H), 3.12-3.21 (m, 1H),
6.90-7.02 (m, 2H), 7.25 (d, J=8.0 Hz, 1H), 7.34 (d, J=7.7 Hz, 1H),
10.62 (s, 1H); MS (APCI.sup.+), m/z 351 (MH.sup.+).
EXAMPLE 5
[0101] 16
[0102]
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)]-13b-Allyl-3,4,4a,5,7,8-
,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-on-
e
[0103] Prepared in 76% yield from
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.bet-
a.)]-13b-allyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4-
]pyrido[1,2-b]-isoquinolin-2-ol by the procedure described in
Example 4. Recrystallization from ethyl acetate/hexane gave product
of mp 220-223.degree. C.; .sup.1H NMR (DMSO-d.sub.6).delta.
1.30-1.38 (m, 1H), 1.48 (t, J=13.0 Hz, 1H), 1.75-1.81 (m, 4H),
1.98-2.21 (m, 3H), 2.33-3.08 (m, 9H), 4.75 (dd, J=2.2, 10.0 Hz,
1H), 4.87 (d, J=17.3 Hz, 1H), 5.45 (m, 1H), 6.85-6.96 (m, 2H), 7.20
(d, J=7.8 Hz, 1H), 7.29 (d, J=7.8 Hz, 1H), 10.59 (s, 1H); MS
(APCI.sup.+), m/z 335 (MH.sup.+).
EXAMPLE 6
[0104] 17
[0105] [2R-(2.alpha.,4a.alpha., 13.beta.,
14a.beta.)]-13b-Propyl-3,4,4a,5,-
7,8,13,13b,14,14a-decahydro-1H-indolo[2',3';3,4]pyrido[1,2-b]isoquinolin-2-
-one
[0106] Prepared in 75% yield from [2R-(2.alpha.,4a.alpha.,
13.beta.,
14a.beta.)]-13b-propyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[-
2',3':3,4]pyrido[1,2-b]-isoquinolin-2-ol by the procedure described
in Example 4. Recrystallization from ethyl acetate/hexane gave
product of mp 255.degree. C. dec.; .sup.1H NMR
(DMSO-d.sub.6).delta. 0.75-0.79 (m, 4H), 1.17-1.39 (m, 2H),
1.56-1.68 (m, 2H), 1.74-1.81 (m, 4H), 2.03-2.26 (m, 4H), 2.38-3.19
(m, 7H), 6.90-7.01 (m, 2H), 7.25 (d, J=7.7 Hz, 1H), 7.33 (d, J=7.7
Hz, 1H), 10.62 (s, 1H); MS (APCI.sup.+), m/z 337 (MH.sup.+).
EXAMPLE 7
[0107] 18
[0108] [4aS-(2E,4a.alpha.,
13.beta.,14a.beta.)]-13b-Allyl-3-benzylidene-3,-
4,4a,5,7,8,13,13b,14,14a-decahydro-.sup.1H-indolo[2',3':3,4]pyrido[1,2-b]i-
soquinolin-2-one A solution of
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)-
]-13b-allyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrid-
o[1,2-b]-isoquinolin-2-one (0.26 g, 0.78 mmol) in 7.0 mL of
methanol and 0.32 mL (0.80 mmol) of 10% aqueous sodium hydroxide
was heated to reflux. A solution of benzaldehyde (0.21 mL, 0.22 g,
2.1 mmol) in 3.0 mL of methanol was added dropwise, and heating was
continued for 4 hours. The cooled reaction mixture was evaporated,
and the residue was partitioned between ethyl acetate and brine.
The organic layer was dried (Na.sub.2SO.sub.4) and evaporated.
Purification of the residue by flash chromatography (25:75:1 ethyl
acetate/hexane/triethylamine) gave 0.27 g (82%) of the product as a
foam; .sup.1H NMR (DMSO-d.sub.6).delta. 0.78-0 98 (m, 1H), 1 14 (s,
2H), 1 50 (t, J=12.1 Hz, 1H), 1.67-1.81 (m, 1H), 1.87-1.95 (m, 1H),
2.08-2.24 (m, 2H), 2.38-2.72 (m, 3H), 2.77-3.21 (m, 5H), 4.83 (d,
J=10.3 Hz, 1H), 4.97 (d, J=17.1 Hz, 1H), 5.45-5.60 (m, 1H),
6.87-7.03 (m, 2H), 7.21-7.53 (m, 8H), 10.67 (s, 1H); MS
(APCI.sup.+), m/z 423 (MH.sup.+).
EXAMPLE 8
[0109] 19
[0110]
[4aS-(2E,4a.alpha.,13.beta.,14a.beta.)]-3-Benzylidine-13b-propyl-3,-
4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoqui-
nolin-2-one
[0111] Prepared in 71% yield from
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.bet-
a.)]-13b-propyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo
[2',3':3,4] pyrido[1, 2-b]-isoquinolin-2-one by the procedure
described in Example 7. The product was obtained as a foam; .sup.1H
NMR (DMSO-d.sub.6).delta.0.71- -0.88 (m, 4H), 1.21-1.36 (m, 1H),
1.58(t, J=12.3 Hz, 1H), 1.63-1.79 (m, 2H), 1.83 (m, 1H), 2.02-2.23
(m, 3H), 2.30-2.70 (m, 4H), 2.78-2.95 (m, 4H), 3.05-3.19 (m, 1H),
6.89-7.01 (m, 2H), 7.11-7.52 (m, 8H), 10.64 (s, 1H); MS
(APCI.sup.+), m/z 425 (MH.sup.+)
EXAMPLE 9
[0112] 20
[0113] [4aS-(2E,4a.alpha.,
13.beta.,14a.beta.)]-3-Benzylidine-13b-butyl-3,-
4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoqui-
nolin-2-one
[0114] Prepared in 31% yield from
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.bet-
a.)]-13b-butyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]py-
rido[1,2-b]-isoquinolin-2-one by the procedure described in Example
7. The product was obtained as a foam; .sup.1H NMR
(DMSO-d.sub.6).delta. 0.64-0.82 (m, 4H), 1.12-1.30 (m, 3H),
1.51-1.84 (m, 4H), 2.03-2.28 (m, 3H), 2.32-2.71 (m, 4H), 2.78-2.95
(m, 4H), 3.08-3.18 (m, 1H), 6.89-7.01 (m, 2H), 7.25-7.50 (m, 8H),
10.64 (s, 1H); MS (APCI.sup.+), m/z 439 (MH.sup.+).
EXAMPLE 10
[0115] 21
[0116]
[2S-(2.alpha.,4a.alpha.,13.beta.,14a.alpha.)]-13b-Allyl-3-benzylide-
ne-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2--
b]isoquinolin-2-ol
[0117] A solution of [4aS-(.sup.2E,4a.alpha.,13.beta.,
14a.beta.)]-13b-allyl-3-benzylidene-3,4,4a,5,7,8,13,13b,14,14a-decahydro--
1H-indolo[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-one (1.8 g, 4.3
mmol) in 75 mL of methanol was cooled in an ice bath and treated
slowly with sodium borohydride (0.73 g, 19.3 mmol). The mixture was
stirred at room temperature for 18 hours. The solvent was
evaporated, and the residue was partitioned between ethyl acetate
and water. The layers were separated, and the aqueous layer was
extracted with fresh ethyl acetate. The combined organic layers
were washed with brine, dried (Na.sub.2SO.sub.4), and evaporated.
The residue was purified by flash chromatography (33:67:0.5 ethyl
acetate/hexane/triethylamine) to yield 1.3 g (72%) of product. A
sample recrystallized from ethyl acetate/hexane had mp
211-213.degree. C.; 1H NMR (DMSO-d.sub.6).delta. 1.09 (q, J=11.5
Hz, 1H), 1.26-1.38 (m, 1H), 1.41 (t, J=12.7 Hz, 1H), 1.64 (t, J=12
9 Hz, 1H), 1.72-1.77 (m, 1H), 1.86-1.92 (m, 2H), 2.55-3.02 (m, 9H),
4.13 (m, 1H), 4.82 (d, J=10.0 Hz, 1H), 4.96 (d, J=17.8 Hz, 1H),
5.10 (d, J=5.1 Hz, 1H), 5.51 (m, 1H), 6.57 (s, 1H), 6.91 (t, J=7.3
Hz, 1H), 6.99 (t, J=7.4 Hz, 1H), 7.18-7.37 (m, 7H), 10.64 (s, 1H);
MS (APCI.sup.+), m/z 425 (MH.sup.+)
EXAMPLE 11
[0118] 22
[0119] [2S-(2.alpha.,4a.alpha.,13.beta.,
14a.beta.)]-3-Benzylidine-13b-pro-
pyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-
-b]-isoquinolin-2-ol
[0120] Prepared in 80% yield from [4aS-(2E,4a.alpha.,13.beta.,
14a.beta.)]-3-benzylidene-13b-propyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-
-1H-indolo[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-one by the
procedure described in Example 10. A sample recrystallized from
ethanol/diethyl ether had mp 224-226.degree. C.; .sup.1H NMR
(DMSO-d.sub.6).delta. 0.76-0.77 (m, 4H), 1.08 (q, J=11.7 Hz, 1H),
1.15-1.35 (m, 2H), 1.46 (t, J=12.5 Hz, 1H), 1.52-1.92 (m, 5H),
2.07-2.20 (m, 1H), 2.52-2.91 (m, 6H), 2.99-3.11 (m, 1H), 4.11 (m,
11), 5.09 (d, J=4.2 Hz, 1H), 6.56 (s, 1H), 6.91 (t, J=7.4 Hz, 1H),
6.98 (t, J=7.5 Hz, 1H), 7.17-7.36 (m, 7H), 10.62 (s, 1H); MS
(APCI.sup.+), m/z 427 (MH.sup.+).
EXAMPLE 12
[0121] 23
[0122]
[2S-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)]-3-Benzylidine-13b-buty-
l-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-1-indolo[2',3':3,4]pyrido[1,2-
-b]-isoquinolin-2-ol
[0123] Prepared in 68% yield from
[4aS-(2E,4a.alpha.,13.beta.,14a.beta.)]--
3-benzylidene-13b-butyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',-
3':3,4]pyrido[1,2-b]-isoquinolin-2-one by the procedure described
in Example 10. A sample recrystallized from aqueous 2-propanol had
mp 222-225.degree. C.; .sup.1H NMR (DMSO-d.sub.6).delta. 0.63-0.81
(m, 4H), 1.02-1.35 (m, 5H), 1.43-1.79 (m, 5H), 1.85-1.93 (m, 1H),
2.15-2.23 (m, 1H), 2.55-2.88 (m, 6H), 3.01-3.12 (m, 1H), 4.05-4.16
(m, 1H), 5.07 (d, J=5.3 Hz, 1H), 6.56 (s, 1H), 6.89-7.00 (m, 2H),
7.17-7.34 (m, 7H), 10.61 (s, 1H); MS (APCI.sup.+), m/z 441
(MH.sup.+).
EXAMPLE 13
[0124] 24
[0125]
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-Benzylidine-10-bromo-13-
b-methyl-3,4,4a,
5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[-
1,2-b]-isoquinolin-2-one
[0126] A solution of
[2R-(2.alpha.,4a.alpha.,13.beta.,14a.beta.)]-13b-meth-
yl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2--
b]-isoquinolin-2-ol (1.5 g, 4.8 mmol; Zinnes H., Comes R, and
Shavel J., Jr., J. Org. Chem, 1965;30:105) in 12 mL of methanol and
12 mL of acetic acid was treated dropwise with a solution of
bromine (0.29 mL, 0.90 g, 5.6 mmol) in 3.0 mL of acetic acid. The
mixture was stirred for 15 minutes, then condensed 50% without
external heating. The residue was treated with 50 mL of diethyl
ether and stirred for 15 minutes. The precipitated solid was
filtered and washed several times with fresh ether. The solid was
partitioned between 100 mL of ammonium hydroxide solution plus 200
mL of water and 150 mL of chloroform. The layers were separated,
and the aqueous layer was extracted with fresh chloroform. The
combined organic layers were washed with brine, dried
(Na.sub.2SO.sub.4), and evaporated to yield 1.9 g of brominated
intermediate as a mixture of isomers.
[0127] The above intermediate was oxidized with dimethyl sulfoxide
and oxalyl chloride following the procedure described in Example 4.
The resulting mixture of ketones was condensed with benzaldehyde
following the procedure described in Example 7. The final product
was purified by flash chromatography (40:60:1 ethyl
acetate/hexane/triethylamine) to yield 0.14 g (6% overall yield) of
the title compound; .sup.1H NMR (DMSO-d.sub.6).delta. 0.73-0.91 (m,
1H), 1.05-1.38 (m, 6H), 1.53-1.70 (m, 1H), 1.90-2.19 (m, 3H),
2.53-2.97 (m, 61), 6.95-7.03 (m, 1H), 7.13-7.50 (m, 8H), 10.89 (s,
1H); MS (APCI.sup.+), m/z 477 (M+2).sup.+.
EXAMPLE 14
[0128] 25
[0129]
[2R-[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]-3-Benzylidine-10-b-
romo-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,-
4]pyrido[1,2-b]-isoquinolin-2-ol
[0130] Prepared in 64% yield from
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]-
]-3-benzylidene-10-bromo-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1-
H-indolo[2',3':3,4]-pyrido[1,2-b]isoquinolin-2-one by the procedure
described in Example 10. The crude product was purified by flash
chromatography (50:50:1 ethyl acetate/hexane/triethylamine) to
yield the title compound; .sup.1H NMR (DMSO-d.sub.6).delta.
0.77-0.92 (m, 1H), 1.12-1.37 (m, 5H), 1.58-1.82 (m, 2H), 1.91-2 07
(m, 2H), 2.48-2.92 (m, 7H), 4.08-4.18 (m, 1H), 5.11 (d, J=5.4 Hz,
1H), 6.57 (s, 1H), 7.04 (d, J=8.6 Hz, 1H), 7.15-7.42 (m, 8H), 10.91
(s, 1H); MS (APCI.sup.+), m/z 479 (M+2).sup.+.
EXAMPLE 15
[0131] 26
[0132] [4aR-[(E),4a.alpha.,13b.beta.,
14a.beta.]]-3-Benzylidine-13b-methyl-
-10-nitro-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[-
1,2-b]-isoquinolin-2-one, and
[0133] [4aR-[(E),4a.alpha.,13b.beta.,
14a.beta.]]-3-Benzylidine-13b-methyl-
-12-nitro-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[-
1,2-b]-isoquinolin-2-one
[0134] A solution of [2R-(2a4a',13.beta.,
14a.beta.)]-13b-methyl-1,2,3,4,4-
a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':3,4]pyrido[1,2-b]-isoquinol-
in-2-ol (1.4 g, 4.5 mmol; Zinnes H., Comes R. A, and Shavel J.,
Jr., J. Org. Chem., 1965;30:105) in 10 mL of acetic acid was
treated dropwise over 30 seconds with a mixture of 2.4 mL of acetic
acid and 2.4 mL (3.8 g, 60 mmol) of fuming nitric acid. The mixture
was stirred for 2 minutes, then poured over 40 g of ice and water.
Concentrated ammonium hydroxide solution was added until the
reaction mixture had a pH of 5.0. A solution of potassium nitrate
was added to precipitate the product as the nitric acid salt. The
aqueous layer was decanted from the precipitated salt, and the
residue was dissolved in methanol. The solution was treated with
ammonium hydroxide to precipitate the nitrated intermediate (0.96
g, 60%) as a mixture of 10- and 12-nitro isomers.
[0135] A solution of the above isomer mixture (0.90 g, 3.1 mmol) in
6.0 mL of dichloromethane was treated with 4-methylmorpholine
N-oxide (0.46 g, 3.9 mmol), tetrapropylammonium perruthenate (0.18
g, 0.51 mmol), and 4A powdered molecular sieves (1.5 g). The
mixture was stirred for 4 hours at room temperature, and the
solvent was evaporated. The residue was purified by flash
chromatography (80:20 chloroform/acetone) to yield 0.45 g (41%) of
an isomeric mixture of nitro ketones.
[0136] A solution of the above isomer mixture (0.44 g, 1.2 mmol) in
1.0 mL of methanol and 6.0 mL of tetrahydrofuiran was treated with
0.6 mL of 10% aqueous sodium hydroxide solution. The mixture was
heated to reflux, and a solution of benzaldehyde (0.29 mL, 0.30 g,
2.9 mmol) in 4.0 mL of tetrahydrofuiran was added dropwise. Heating
at reflux was continued for 16 hours. The cooled reaction mixture
was evaporated, and the residue was partitioned between ethyl
acetate and brine. The organic layer was dried (MgSO.sub.4) and
evaporated. Purification of the residue by flash chromatography
(60:30: 10 ethyl acetate/hexane (triethylamine) gave 0.078 g (14%)
of [4aR-[(E),4a.alpha., 13b.beta., 14a.beta.]]-3-benzylidine-13b--
methyl-10-nitro-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]p-
yrido[1,2-b]-isoquinolin-2-one; .sup.1H NMR (DMSO-d.sub.6).delta.
0.75-0.97 (m, 1H), 1.08-1.15 (m, 2H), 1.32 (s, 3H), 1.35-1.47 (m,
1H), 1.58-1.80 (m, 1H), 2.03-2.22 (m, 3H), 2.40-2.91 (m, 6H),
7.10-7.52 (m, 7H), 7.80 (d, J=8.8 Hz, 1H), 8.13 (s, 1H), 11.61 (s,
1H); MS (APCI.sup.+), m/z 442 (and 0.066 g (12%) of
[4aR-[(E),4a.alpha.,13b.beta.-
,14a.beta.]]-3-benzylidine-13b-methyl-12-nitro-3,4,4a,5,7,8,13,13b,14,14a--
decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-one;
.sup.1H NMR (DMSO-d.sub.6).delta. 0.75-0.88 (m, 1H), 1.04-1.45 (m,
6H), 1.58-1.89 (m, 2H), 1.96-2.28 (m, 2H), 2.57-2.93 (m, 4H),
3.07-3.18 (m, 2H), 7.06-7.11 (m, 1H), 7.30-7.52 (m, 5H), 7.61-7.83
(m, 2H), 8.12 (s, 1H), 11.68 (s, 1H); MS (APCI.sup.+), m/z 442
(MH.sup.+).
EXAMPLE 16
[0137] 27
[0138]
[2R-[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]-3-Benzylidine-13b--
methyl-10-nitro-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3:3,4-
]pyrido[1,2-b]-isoquinolin-2-ol
[0139] A solution of
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-benzylide-
ne-13b-methyl-10-nitro-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3-
':3,4]pyrido[1,2-b]-isoquinolin-2-one (0.071 g, 0.16 mmol) in 5.0
mL of tetrahydrofuran and 1.0 mL of methanol was cooled in an ice
bath and treated slowly with sodium borohydride (0.031 g, 0.82
mmol). The mixture was stirred at room temperature for 16 hours,
then cooled in ice and treated with 1.0 mL of water and 4.0 mL of
acetic acid. The pH of the mixture was adjusted to 9.0 by the
addition of ammonium hydroxide solution. The mixture was extracted
with chloroform, and the combined extracts were washed with brine,
dried (MgSO.sub.4), and evaporated. The residue was recrystallized
from methanol to yield 0.048 g (68%) of product; .sup.1H NMR
(CDCl.sub.3).delta. 1.12-1.25 (m, 1H), 1.39 (s, 3H), 1.40-1.78 (m,
4H), 1.83-1.92 (m, 1H), 2.00 (s, 1H), 2.04-2.13 (m, 1H), 2.52-3.05
(m, 7H), 4.21-4.32 (m, 1H), 6.61 (s, 1H), 7.08-7.35 (m, 5H), 7.42
(d, J=8.6 Hz, 1H), 7.95 (m, 1H), 8.23 (s, 1H), 8.45 (s, 1H); MS
(APCI.sup.+), m/z 444 (MH.sup.+).
EXAMPLE 17
[0140] 28
[0141] [2R-[2.alpha.(E),4a.alpha., 13b.beta.,
14a.beta.]]-3-Benzylidine-13-
b-methyl-12-nitro-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3':-
3,4]pyrido[1,2-b]-isoquinolin-2-ol
[0142] Prepared in 29% yield from [4aR-[(E),4a.alpha.,13b.beta.,
14a.beta.]]-3-benzylidene-13b-methyl-12-nitro-3,4,4a,5,7,8,13,13b,14,14a--
decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-one by the
procedure described in Example 16. The final product was
recrystallized from methanol; .sup.1H NMR (CDCl.sub.3).delta.
1.11-1.23 (m, 1H), 1.35 (s, 3H), 1.38-2.14 (m, 7H), 2.43-2.69 (m,
2H), 2.73-2.92 (m, 3H), 3.03-3.16 (m, 1H), 3.43 (s, 1H), 4.20-4.30
(m, 1H), 6.57 (s, 1H), 7.02-7.33 (m, 6H), 7.49 (d, J=8.1 Hz, 1H),
7.82 (d, J=8.1 Hz, 1H), 8.08 (s, 1H); MS (APCI.sup.+), m/z 444
(MH.sup.+).
EXAMPLE 18
[0143] 29
[0144] [4aR-[(E),4a.alpha., 13b.beta.,
14a.beta.]]-3-(3,4-Dimethoxy-benzyl-
idene)-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4-
]pyrido[1,2-b]-isoquinolin-2-one
[0145] To a solution of [4aR-(4a.alpha., 13b.beta.,
14a.beta.)]-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',-
3.alpha.:3,4]pyrido[1,2-b]-isoquinolin-2-one (0.25 g, 0.81 mmol;
Zinnes H., Comes R. A., and Shavel J., Jr., J. Org. Chem.,
1965;30:105) in 15 mL of methanol was added
3,4-dimethoxybenzaldehyde (0.61 g, 3.7 mmol), followed by 0.30 mL
of 10% aqueous sodium hydroxide solution. The mixture was stirred
at reflux for 18 hours. The solvent was evaporated, and the residue
was dissolved in a minimum amount of dichloromethane and purified
by flash chromatography (70:30 ethyl acetateihexane) to yield 0.17
g (47%) of the benzylidene ketone of mp 120-125.degree. C.; .sup.1H
NMR (CDCl.sub.3).delta. 1.34 (s, 3H), 1.54 (t, J=12.1 Hz, 1H),
1.70-1.88 (m, 1H), 1.88-2.15 (m, 3H), 2.27-2.39 (m, 1H), 2.54-2.76
(m, 4H), 2.79-3.04 (m, 4H), 3.82 (s, 3H), 3.85 (s, 3H), 6.84 (d,
J=8.4 Hz, 1H), 6.88 (s, 1H), 6.96-7.09 (m, 3H), 7.23 (d, J=7.9 Hz,
1H), 7.39 (d, J=7.5 Hz, 1H), 7.47 (s, 1H), 7.81 (s, 1H); MS
(APCI.sup.+), m/z 457 (MH.sup.+).
EXAMPLE 19
[0146] 30
[0147] [4aR-[(E),4a.alpha.,
13b.beta.,14a.beta.]]3-(3-Methoxy-benzylidene)- -13b-methyl-3,4,4a,
5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyri-
do[1,2-b]-isoquinolin-2-one
[0148] Prepared in 46% yield from [4aR-(4a.alpha.,13b.beta.,
14a.beta.)]-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',-
3':3,4]pyrido[1,2-b]-isoquinolin-2-one and 3-methoxybenzaldehyde by
the procedure described in Example 18. The product had mp
125-130.degree. C.; .sup.1H NMR (CDCl.sub.3).delta. 1.10-1.25 (m,
1H), 1.35 (s, 3H), 1.45-3.05 (m, 3H), 3.75 (s, 3H), 6.75-7.50 (m,
9H), 7.75 (s, 1H); MS (APCI.sup.+), m/z 427 (MH.sup.+).
EXAMPLE 20
[0149] 31
[0150] [4aR-[(E),4a.alpha.,13b.beta.,
14a.beta.]]-3-(3,5-Dimethoxy-benzyli-
dene)-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]-
pyrido[1,2-b]-isoquinolin-2-one
[0151] Prepared in 44% yield from [4aR-(4a.alpha., 13b.beta.,
14a.beta.)]-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',-
3':3,4]pyrido[1,2-b]-isoquinolin-2-one and
3,5-dimethoxybenzaldehyde by the procedure described in Example 18.
The product had mp 110-120.degree. C.; .sup.1H NMR
(CDCl.sub.3).delta. 1.37 (s, 3H), 1.47-1.69 (m, 1H), 1.73-1.89 (m,
1H), 1.91-2.25 (m, 3H), 2.25-2.41 (m, 1H), 2.56-2.79 (m, 4H),
2.79-3.14 (m, 4H), 3.78 (s, 6H), 6.43 (s, 1H) 6.50 (s, 2H),
6.99-7.20 (m, 2H), 7.27 (d, J=8.0 Hz, 1H), 7.42 (s, 2H), 7.66 (s,
1H), MS (APCI.sup.+), m/z 457 (MH.sup.+).
EXAMPLE 21
[0152] 32
[0153] [4aR-[(E),4a.alpha.,13b.beta.,
14a.beta.]]-3-(3,4-Dichloro-benzylid-
ene)-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]p-
yrido[1,2-b]-isoquinolin-2-one
[0154] Prepared in 17% yield from
[4aR-(4a.alpha.,13b.beta.,14a.beta.)]-13-
b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1-
,2-b]-isoquinolin-2-one and 3,4-dichlorobenzaldehyde by the
procedure described in Example 18. The product had mp
140-145.degree. C.; .sup.1H NMR (CDCl.sub.3).delta. 1.34 (s, 3H),
1.57 (t, J=12.0 Hz, 1H), 1.71-1.88 (m, 1H), 1.88-2.17 (m, 3H),
2.21-2.37 (m, 1H,), 2.54-2.78 (m, 4H), 2.78-3.05 (m, 4H), 7.02 (t,
J=7.6 Hz, 1H), 7.08 (t, J=7.2 Hz, 1), 7.15 (dd, J=8.6, 1.7 Hz, 1H),
7.24 (d, J=7.9 Hz, 1H), 7.30-7.46 (m, 4H), 7.61 (s, 1H); MS
(APCI.sup.+), m/z 465 (MH.sup.+ ).
EXAMPLE 22
[0155] 33
[0156] [2R-[2.alpha.(E),4a.alpha., 13b.beta.,
14a.beta.]]-3-(3,4-Dimethoxy-
-benzylidene)-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo-
[2',3':3,4]pyrido[1,2-b]-isoquinolin-2-ol
[0157] A solution of
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-(3,4-dime-
thoxy-benzylidene)-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indo-
lo[2',3':3,4]-pyrido[1,2-b]-isoquinolin-2-one (0.15 g, 0.34 mmol)
in 2.0 mL of methanol and 4.0 mL of tetrahydrofuiran was treated
with potassium borohydride (0.09 g, 1.7 mmol). The mixture was
stirred at room temperature for 18 hours. The solvent was
evaporated, and the residue was suspended in 3.0 mL of water and
treated with 4.0 mL of glacial acetic acid. The solution was
allowed to-stir at room temperature for 30 minutes, and then 5.0 mL
of ammonium hydroxide was added until pH 8. The precipitated
product was extracted with chloroform. The organic layer was dried
(Na.sub.2SO.sub.4) and evaporated to yield an oil. The residue was
purified by flash chromatography (95:5 chloroform/methanol) to
yield 0.10 g (68%) of product of mp 118-121.degree. C.; .sup.1H NMR
(CDC1.sub.3).delta. 1.08-1.17 (m, 1H), 1.31 (s, 3H), 1.37-1.76 (m,
4H), 1.79 (dd, J=12.1, 3.3 Hz, 1H), 2.01-2.10 (m, 1H), 2.11 (s,
1H), 2.50-2.70 (m, 3H), 2.76-2.98 (m, 4H), 3.80 (s, 3H), 3.82 (s,
3H), 4.22 (dd, J=11.3, 3.5 Hz, 1H), 6.50 (s, 1H), 6.61-6.73 (m,
2H), 6.73-6.82 (d, J=8.3 Hz, 1H), 7.01 (t, J=7.6 Hz, 1H), 7.07 (t,
J=7.6 Hz, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.39 (d, J=8.0 Hz, 1H), 7.61
(s, 1H); MS (APCI.sup.+), m/z 459 (ME.sup.+).
EXAMPLE 23
[0158] 34
[0159] [2R-[2.alpha.((E),4a.alpha., 13b.beta.,
14a.beta.]]-3-(3-Methoxy-be-
nzylidene)-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2'-
,3':3,4]pyrido[1,2-b]-isoquinolin-2-ol
[0160] Prepared in 21% yield from
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]-
]-3-(3-methoxy-benzylidene)-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydr-
o-1H-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-one by the
procedure described in Example 22. The product had mp
140-142.degree. C.; .sup.1H NMR (CDCl.sub.3).delta. 1.16 (m, 1H),
1.35 (s, 3H), 1.40-1.70 (m, 4H), 1.75 (dd, J=12.0, 3.3 Hz, 1H),
2.00-2.10 (m, 2H), 2.54-2.74 (m, 3H), 2.80-3.03 (m, 41 CH.sub.2),
3.80 (s, 3H), 4.25 (dd, J=11.2, 4.3 Hz, 1H), 6.61 (s, 1H),
6.70-6.87 (m, 3H), 7.04-7.19 (m, 2H), 7.19-7.28 (m, 1H), 7.31 (d,
J=7.9 Hz, 1H), 7.47 (d, J=7.7 Hz, 1H), 7.75 (s, 1H); MS
(APCI.sup.+), m/z 429 (MH.sup.+).
EXAMPLE 24
[0161] 35
[0162]
[2R-[2.alpha.(E),4a.alpha.,13b.beta.,14a.beta.]]-3-(3,5-Dimethoxy-b-
enzylidene)-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2-
',3':3,4]pyrido[1,2-b]-isoquinolin-2-ol
[0163] Prepared in 50% yield from
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]-
]-3-(3,5-dimethoxy-benzylidene)-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-deca-
hydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-one by the
procedure described in Example 22. The product had mp
128-132.degree. C.; .sup.1H NMR (CDCl.sub.3).delta. 1.10-1.19 (m,
1H), 1.33 (s, 3H), 1.38-2.00 (m, 5H), 2.02-2.12 (m, 1H), 2.14 (s,
1H), 2.50-2.74 (m, 3H), 2.75-3.04 (m, 4H), 3.75 (s, 6H), 4.24 (dd,
J=11.3, 3.5 Hz, 1H), 6.32 (s, 3H), 6.53 (s, 1H), 7.04 (t, J=7.7 Hz,
1H), 7.10 (t, J=7.4 Hz, 1H), 7.26 (d, J=8.3 Hz, 1H), 7.42 (d, J=7.5
Hz, 1H), 7.65 (s, 1H); MS (APCI.sup.+), m/z 459 (MH.sup.+).
EXAMPLE 25
[0164] 36
[0165] [2R-[2.alpha.(E),4a.alpha.,13b.beta.,
14a.beta.]]-3-(3,4-Dichloro-b-
enzylidene)-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2-
',3':3,4]pyrido[1,2-b]-isoquinolin-2-ol
[0166] Prepared in 44% yield from
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]-
]-3-(3,4-dichloro-benzylidene)-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decah-
ydro-1H-indolo[2',3':3,4]pyrido[1,2-b]isoquinolin-2-one by the
procedure described in Example 22. The product had mp
125-130.degree. C.; .sup.1H NMR (CDCl.sub.3).delta. 1.06-1.17 (m,
1H), 1.31 (s, 3H), 1.34-1.86 (m, 5H), 2.03-2.10 (m, 1H), 2.11 (s,
1H), 2.50-2.70 (m, 3H), 2.76-2.99 (m, 4H), 4.22 (dd, J=11.7, 3.8
Hz, 1H), 6.47 (s, 1H), 6.94 (dd, J=8.2, 1.9 Hz, 1H), 7.01 (t, J=7.7
Hz, 1H), 7.07 (t, J=8.0 Hz, 1H), 7.16-7.27 (m, 2H), 7.32 (d, J=8.3
Hz, 1H), 7.39 (d, J=7.3 Hz, 1H), 7.59 (s, 1H); MS (APCI.sup.+), m/z
467 (MH.sup.+).
EXAMPLE 26
[0167] 37
[0168]
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-Benzylidene-13b-methyl--
3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]-iso-
quinolin-2-one
[0169] Prepared in 50% yield from
[4aR-(4a.alpha.,13b.beta.,14a.beta.)]-13-
b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1-
,2-b]-isoquinolin-2-one and benzaldehyde by the procedure described
in Example 18. The product had mp 135-145.degree. C.; .sup.1H NMR
(CDCl.sub.3).delta. 1.38 (s, 3H), 1.53-1.65 (m, 1H), 1.75-1.89 (m,
1H), 1.92-2.18 (m, 3H), 2.29-2.43 (m, 1H), 2.56-2.79 (m, 4H),
2.79-3.09 (m, 4H), 7.01-7.15 (m, 2H), 7.20-7.49 (m, 7H), 7.52 (s,
1H), 7.67 (s, 1H); MS (APCI.sup.+), m/z 397 (MH.sup.+).
EXAMPLE 27
[0170] 38
[0171]
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-Benzylidene-13b-methyl--
3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1
,2-b]-isoquinolin-2-one oxime
[0172] A solution of
[4aR-[(E),4.alpha.,13b.beta.,14a.alpha.]]-3-benzylide-
ne-13b-methyl-3,4,4a,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyr-
ido[1,2-b]-isoquinolin-2-one (0.12 g, 0.31 mmol) in 4.0 mL of
ethanol and 4.0 mL of water was treated with hydroxylamine
hydrochloride (0.13 g, 1.9 mmol), followed by sodium acetate
hydrate (0.25 g, 1.9 mmol). The mixture was stirred at reflux for
18 hours. The solvent was evaporated, and the residue was suspended
in water and extracted with chloroform. The organic layer was dried
(Na.sub.2SO.sub.4) and evaporated to afford a solid. The crude
product was purified by flash chromatography (40:60 ethyl
acetate/hexane) to yield 0.067 g (53%) of product of mp
142-145.degree. C.; .sup.1H NMR (CDCl.sub.3).delta. 1 32 (s, 3H),
1.47-3.33 (m, 15H), 6.87 (s, 1H), 7.01 (t, J=7.8 Hz, 1H), 7.07 (t,
J=7.2 Hz, 1H), 6.95-7.12 (m, 6H), 7.39 (d, J=7.6 Hz, 1H), 7.61 (s,
1H); MS (APCI.sup.+), m/z 412 (MH.sup.+)
EXAMPLE 28
[0173] 39
[0174]
3-Benzylidene-13b-methyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-
-indolo[2',3': 3,4]pyrido[1,2-b]isoquinolin-2-ylamine zA solution
of
[4aR-[(E),4a.alpha.,13b.beta.,14a.beta.]]-3-benzylidene-13b-methyl-3,4,4a-
,5,7,8,13,13b,14,14a-decahydro-1H-indolo[2',3':3,4]pyrido[1,2-b]-isoquinol-
in-2-one oxime (0.12 g, 0.29 mmol) in 5.0 mL of ethanol and 5.0 mL
of glacial acetic acid was cooled to 0.degree. C. and treated with
zinc dust (0.19 g, 2.9 mmol). The mixture was warmed to room
temperature and stirred for 3 hours. The mixture was filtered, and
the zinc was washed with fresh ethanol. The solvent was evaporated,
and the residue was suspended in 10.0N sodium hydroxide solution
and extracted with chloroform. The organic layer was separated,
dried (Na.sub.2SO.sub.4), and the solvent was evaporated to afford
a solid. The crude product was purified by preparative BPLC
(acetonitrile/water+0.05% trifluoroacetic acid gradient elution) to
yield 0.025 g (14%) of final product as the di-trifluoroacetic acid
salt; mp 203-210.degree. C.; MS (APCI.sup.+), m/z 398
(MH.sup.+).
[0175] The following were prepared by the above procedures:
[0176]
[1R-[1.alpha.,2.beta.(E),4a.beta.,13b.alpha.,14a.alpha.]]3-Benzylid-
ene-1,13b-dimethyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-indolo[2',3'-
:3,4]pyrido] 1,2-b]isoquinolin-3-ol;
[0177]
[1R-[1.alpha.,2.beta.(E),4a.beta.,13b.alpha.,14a.alpha.]]3-Biphenyl-
-4-ylmethylene-1,13b-dimethyl-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-i-
ndolo[2',3':3,4]pyrido]1,2-b]isoquinolin-2-ol; and
[0178]
[1R-[1.alpha.,2.beta.(E),4a.beta.,13b.alpha.,14a.alpha.]]1,13b-Dime-
thyl-3-(4-styryl-benzylidene)-1,2,3,4,4a,5,7,8,13,13b,14,14a-dodecahydro-i-
ndolo[2',3':3,4]pyrido]1,2-b]isoquinolin-2-ol.
* * * * *