U.S. patent application number 09/797081 was filed with the patent office on 2002-02-14 for compounds useful for inhibition of farnesyl protein transferase.
Invention is credited to Alvarez, Carmen, Doll, Ronald J., Lalwani, Tarik, Njoroje, FGeorge, Remiszewski, Stacy W., Taveras, Arthur G..
Application Number | 20020019400 09/797081 |
Document ID | / |
Family ID | 26699501 |
Filed Date | 2002-02-14 |
United States Patent
Application |
20020019400 |
Kind Code |
A1 |
Njoroje, FGeorge ; et
al. |
February 14, 2002 |
Compounds useful for inhibition of farnesyl protein transferase
Abstract
Novel compounds of the formula: 1 are disclosed. Compounds of
Formula 1.0 are represented by the compounds of formulas: 2 wherein
R.sup.1, R.sup.3 and R.sup.4 are each independently selected from
halo. Also disclosed are methods of inhibiting farnesyl protein
transferase and the growth of abnormal cells, such as tumor
cells.
Inventors: |
Njoroje, FGeorge; (Union,
NJ) ; Taveras, Arthur G.; (Rockaway, NJ) ;
Doll, Ronald J.; (Maplewood, NJ) ; Lalwani,
Tarik; (Edison, NJ) ; Alvarez, Carmen;
(Roselle Park, NJ) ; Remiszewski, Stacy W.;
(Washington Township, NJ) |
Correspondence
Address: |
SCHERING-PLOUGH CORPORATION
PATENT DEPARTMENT (K-6-1, 1990)
2000 GALLOPING HILL ROAD
KENILWORTH
NJ
07033-0530
US
|
Family ID: |
26699501 |
Appl. No.: |
09/797081 |
Filed: |
March 1, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09797081 |
Mar 1, 2001 |
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09417885 |
Oct 14, 1999 |
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6228856 |
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60025249 |
Sep 13, 1996 |
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60050009 |
Jun 17, 1997 |
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Current U.S.
Class: |
514/253.03 ;
514/290; 544/361; 546/93 |
Current CPC
Class: |
A61P 35/00 20180101;
C07D 221/16 20130101; C07D 401/04 20130101; C07D 221/06 20130101;
C07D 401/14 20130101; C07D 401/12 20130101 |
Class at
Publication: |
514/253.03 ;
514/290; 544/361; 546/93 |
International
Class: |
A61K 031/497; A61K
031/473; C07D 41/08 |
Claims
What is claimed is:
1. A compound of the formula: 321or a pharmaceutically acceptable
salt or solvate thereof, wherein: one of a, b, c and d represents N
or NR.sup.9 wherein R.sup.9 is O--, --CH.sub.3 or
--(CH.sub.2).sub.nCO.sub.2H wherein n is 1 to 3, and the remaining
a, b, c and d groups represent CR.sup.1 or CR.sup.2; or each of a,
b, c, and d are independently selected from CR.sup.1 or CR.sup.2;
each R.sup.1 and each R.sup.2 is independently selected from H,
halo, --CF.sub.3, --OR.sup.10, --COR.sup.10, --SR.sup.10,
--S(O).sub.tR.sup.11 (wherein t is 0, 1 or 2), --SCN,
--N(R.sup.10).sub.2, --NR.sup.10R.sup.11, --NO.sub.2,
--OC(O)R.sup.10, --CO.sub.2R.sup.10, --OCO.sub.2R.sup.11, --CN,
--NHC(O)R.sup.10, --NHSO.sub.2R.sup.10, --CONHR.sup.10,
--CONHCH.sub.2CH.sub.2OH, --NR.sup.10COOR.sup.11,
322--SR.sup.11C(O)OR.sup.11, --SR.sup.11N(R.sup.75).sub.2 wherein
each R.sup.75 is independently selected from H and --C(O)OR.sup.11,
benzotriazol-1-yloxy, tetrazol-5-ylthio, or substituted
tetrazol-5-ylthio, alknyl, alkenyl or alkyl, said alkyl or alkenyl
group optionally being substituted with halo, --OR.sup.10 or
--CO.sub.2R.sup.10; R.sup.3 and R.sup.4 are the same or different
and each independently represents H, any of the substituents of
R.sup.1 and R.sup.2, or R.sup.3 and R.sup.4 taken together
represent a saturated or unsaturated C5-C7 fused ring to the
benzene ring (Ring III); R.sup.5, R.sup.6, R.sup.7 and R.sup.8 each
independently represents H, --CF.sub.3, --COR.sup.10, alkyl or
aiyl, said alkyl or aiyl optionally being substituted with
--OR.sup.10, --SR.sup.10, --S(O).sub.tR.sup.11,
--NR.sup.10COOR.sup.11, --N(R.sup.10).sub.2, --NO.sub.2,
--COR.sup.10, --OCOR.sup.10, --OCO.sub.2R.sup.11,
--CO.sub.2R.sup.10, OPO.sub.3R.sup.10, or R.sup.5 is combined with
R.sup.6 to represent .dbd.O or .dbd.S and/or R.sup.7 is combined
with R.sup.8 to represent .dbd.O or .dbd.S; R.sup.10 represents H,
alkyl, aryl, or aralkyl; R.sup.11 represents alkyl or aryl; X
represents N, CH or C, which C may contain an optional 10 double
bond (represented by the dotted line) to carbon atom 11; the dotted
line between carbon atoms 5 and 6 represents an optional double
bond, such that when a double bond is present, A and B
independently represent --R.sup.10, halo, --OR.sup.11,
--OCO.sub.2R.sup.11 or --OC(O)R.sup.10, and when no double bond is
present between carbon atoms 5 and 6, A and B each independently
represent H.sub.2, --(OR.sup.11).sub.2; H and halo, dihalo, alkyl
and H, (alkyl).sub.2, --H and --OC(O)R.sup.10, H and --OR.sup.10,
.dbd.O, aryl and H, .dbd.NOR.sup.10 or --O--(CH.sub.2).sub.p--O--
wherein p is 2, 3 or 4; and W represents a group selected from the
group consisting of: 323wherein: R.sup.12 is selected from the
group consisting of: (a) H; (b) alkyl; (c) aralkyl; and (d)
heteroarylalkyl; R.sup.13 and R.sup.14 are each independently
selected from the group consisting of: (a) H; (b) --C(O)OR.sup.16
wherein R.sup.16 represents alkyl, aralkyl, and heteroaralkyl; (c)
--SO.sub.2R.sup.17 wherein R.sup.17 is selected from the group
consisting of: NH.sub.2, --N(alkyl).sub.2 wherein each alkyl is the
same or different, alkyl, aryl, aralkyl, heteroaryl and
heteroaralkyl; (d) --C(O)R.sup.18 wherein R.sup.18 is selected from
the group consisting of: aryl, alkyl, aralkyl, heteroaryl, and
heteroaralkyl; (e) C.sub.1-6 alkyl; (f) alkaryl; and (g) C.sub.3-6
cycloalkyl; r is0, 1 or 2; s represents 1, 2, 3, 4, or 5, and each
Y for each --CY.sub.2-- group is independently selected from H or
--OH, provided that both Y substituents of each --CY.sub.2-- group
are not --OH, and provided that for the --CY.sub.2-- group alpha to
the nitrogen both Y substituents are H, such that the group
324forms a 3, 4, 5, 6, or 7 membered ring; v is 0, 1 or 2; R.sup.15
is selected from the group consisting of: (a) heteroaryl; (b) a
group selected from: 325(c) heterocycloalkyl selected from the
group consisting of: 326z is 0, 1, 2, 3, 4, or 5 wherein each
--CH.sub.2-- group is optionally substituted with a --OH group;
R.sup.22 represents a group selected from: 327(5) alkyl (e.g.,
--CH.sub.3), (6) --OR.sup.23 wherein R.sup.23 is selected from the
group consisting of: alkyl, aryl and H, and 328wherein R.sup.24 and
R.sup.25 are independently selected from the group consisting of:
--NH.sub.2, alkoxy, --OH, --CH.sub.2CO.sub.2H, --OCH.sub.2Ph,
--CH(OCH.sub.3)CH(CH.sub.3).sub- .2, alkyl, aryl, H, aralkyl, and
heteroaralkyl; or R.sup.24 and R.sup.25 taken together form a
carbon chain having 4 or 5 (--CH.sub.2--) groups such that R.sup.24
and R.sup.25 taken together with the nitrogen to which they are
bound form a 5 or 6 membered heterocycloalkyl ring.
2. The compound of claim 1 wherein R.sup.2 is H; R.sup.1 is
selected from the group consisting of: Br and Cl; R.sup.3 is
selected from the group consisting of: Br and Cl; R.sup.4 is
selected from the group consisting of: H, Br and Cl; R.sup.5,
R.sup.6, R.sup.7 and R.sup.8 are H; A and B are each H.sub.2; and
the optional bond between C5 and C6 is absent.
3. The compound of claim 2 wherein R.sup.4 is H.
4. The compound of claim 2 wherein R.sup.4 is selected from the
group consisting of: Cl or Br.
5. The compound of claim 4 wherein W is selected from the group
consisting of: 329wherein: (1) r is 0; (2) R.sup.12 is selected
from the group consisting of: (a) H; (b) alkyl; (c) aralkyl; and
(d) heteroaralkyl; and (3) R.sup.13 and R.sup.14 are independently
selected from the group consisting of: (a) H; (b) --C(O)OR.sup.16
wherein R.sup.16 is alkyl: (c) --SO.sub.2R.sup.17 wherein R.sup.17
is alkyl or aryl; (d) --C(O)R.sup.18 wherein R.sup.18 is aryl; and
(e) alkyl; 330wherein: (1) r is 1 or2: (2) R.sup.12 is H; and (3)
R.sup.13 is alkyl and R.sup.14 is H, alkyl or --C(O)OR.sup.16
wherein R.sup.16 is alkyl; 331wherein: (1) s is 1,2, 3,4 or 5; and
(2) R.sup.13 is selected from the group consisting of: (a) H and
--C(O)OR.sup.16 wherein R.sup.16 is alkyl; 332wherein: (1) v is 0;
(2) R.sup.12 is H; and (3) R.sup.15 is selected from the group
consisting of: 333 and --OH, --CN; 334wherein: (1) v is 1 or2; (2)
R.sup.12 is H; and (3) R.sup.15 is heterocycloalkyl; 335wherein:
(1) z is 0; (2) R.sup.22 is --NR.sup.24R.sup.25; and (3) R.sup.24
and R.sup.25 are independently selected from: H, --NH.sub.2, alkyl,
alkoxy, --OH, --CH.sub.2CO.sub.2H, or --OCH.sub.2C.sub.6H.sub.5;
and 336wherein: (1) z is 1, 2, 3, 4 or 5; (2) R.sup.22 is selected
from: --OR.sup.23, --ONa, --OLi, alkyl, --NR.sup.24R.sup.25 or
337(3) R.sup.23 is alkyl; and (4) R.sup.24 and R.sup.25 are
independently selected from H, --CH(OCH.sub.3)CH(CH.sub.3).sub.2,
338
6. The compound of claim 5 wherein X is CH.
7. The compound of claim 6 wherein R.sup.1 is Br, R.sup.3 is Cl and
R.sup.4 is Br.
8. The compound of claim 1 selected from: 339wherein R.sup.1,
R.sup.3 and R.sup.4 are each independently selected from halo; and
A, B, X and W are as defined in claim 1.
9. The compound of claim 8 wherein R.sup.1 is selected from the
group consisting of: Br and Cl; R.sup.3 and R.sup.4 are
independently selected from the group consiting of: Br and Cl: A
and B are each H.sub.2; and the optional bond between C5 and C6 is
absent.
10. The compound of claim 9 wherein R.sup.1 is Br; R.sup.3 is Cl;
and R.sup.4 is Br.
11. The compound of claim 10 wherein W is selected from the group
consisting of: 340wherein: (1) r is 0; (2) R.sup.12 is selected
from the group consisting of: (a) H; (b) alkyl; (c) aralkyl; and
(d) heteroaralkyl; and (3) R.sup.13 and R.sup.14 are independently
selected from the group consisting of: (a) H; (b) --C(O)OR.sup.16
wherein R.sup.16 is alkyl; (c) --SO.sub.2R.sup.17 wherein R.sup.17
is alkyl or aryl; (d) --C(O)R.sup.18 wherein R.sup.18 is aryl; and
(e) alkyl; 341wherein: (1) r is 1 or2; (2) R.sup.12 is H; and (3)
R.sup.13 is alkyl and R.sup.14 is H, alkyl or --C(O)OR.sup.16
wherein R.sup.16 is alkyl; 342wherein: (1) s is 1, 2, 3, 4 or 5;
and (2) R.sup.13 is selected from the group consisting of: (a) H
and --C(O)OR.sup.16 wherein R.sup.16 is alkyl; 343wherein: (1) v is
0; (2) R.sup.12 is H; and (3) R.sup.15 is selected from the group
consisting of: 344 345 and --OH, --CN; 346wherein: (1) v is 1 or 2;
(2) R.sup.12 is H; and (3) R.sup.15 is heterocycloalkyl;
347wherein: (1) z is 0; (2) R.sup.22 is --NR.sup.24R.sup.25; and
(3) R.sup.24 and R.sup.25 are independently selected from: H,
--NH.sub.2, alkyl, alkoxy, --OH, --CH.sub.2CO.sub.2H, or
--OCH.sub.2C.sub.6H.sub.5; and 348wherein: (1) z is 1, 2, 3, 4 or
5; (2) R.sup.22 is selected from: --OR.sup.23, --ONa, --OLi, alkyl,
--NR.sup.24R.sup.25 or 349(3) R.sup.23 is alkyl; and (4) R.sup.24
and R.sup.25 are independently selected from H,
--CH(OCH.sub.3)CH(CH.sub.3).sub.2, 350
12. The compound of claim 11 wherein X is CH.
13. The compound of claim 12 wherein W is selected from the group
consisting of: 351wherein: (1) r is 0; (2) R.sup.12 is selected
from the group consisting of: (a) H; (b) methyl; (c)
--CH.sub.2-imidazolyl; and (d) benzyl; and (3) R.sup.13 and
R.sup.14 are independently selected from the group consisting of:
(a) H; (b) --C(O)OC(CH.sub.3).sub.3; (c) --SO.sub.2CH.sub.3; (d)
--C(O)-phenyl; and (e) methyl; 352wherein: (1) r is 1 or 2; (2)
R.sup.12 is H; and (3) R.sup.13 is methyl and R.sup.14 is methyl;
353wherein: (1) s is 3;and (2) R.sup.13 is selected from the group
consisting of: (a) H and --C(O)OC(CH.sub.3).sub.3; 354wherein: (1)
v is 0; (2) R.sup.12 is H; and (3) R.sup.15 is selected from the
group consisting of: 355wherein: (1) v is 1 or 2; (2) R.sup.12 is
H; and (3) R.sup.15 is 356wherein: (1) z is 0; (2) R.sup.22 is
--NR.sup.24R.sup.25; and (3) R.sup.24 and R.sup.25 are each H; and
357wherein: (1) z is 1, 2 or 3; (2) R.sup.22 is --OR.sup.23; and
(3) R.sup.23 is methyl or ethyl.
14. The compound of claim 12 wherein said compound is a compound of
the formula: 358
15. The compound of claim 1 wherein W is 359r is 0 and said
compound is selected from the group consisting of: 360
16. The compound of claim 1 wherein W is 361r is 1 or 2 and is
selected from the group consisting of: 362
17. The compound of Formula 1.0 wherein W is 363s is 3 and said
compound is selcted from the group consisting of: 364
18. The compound of Formula 1.0 wherein W is 365v is 0 and said
compound is selected from the group consisting of: 366
19. The compound of claim 1 wherein W is 367v is 1 and said
compound is selected from the group consisting of: 368
20. The compound of claim 1 wherein W is 369z is 0 and said
compound is selected from the group consisting of: 370
21. The compound of claim 1 wherein W is 371z is 1, 2,3, 4 or 5 and
said compound is selected from the group consisting of: 372
22. A compound selected from the group consisting of: 373or
pharmaceutically acceptable salts or solvates thereof.
23. A method of treating tumor cells expressing an activated ras
oncogene comprising administering an effective amount of a compound
of claim 1.
24. The method of claim 23 wherein the cells treated are pancreatic
tumor cells, lung cancer cells, myeloid leukemia tumor cells,
thyroid follicular tumor cells, myelodysplastic tumor cells,
epidermal carcinoma tumor cells, bladder carcinoma tumor cells or
colon tumors cells.
25. The method of claim 24 wherein the treatment is of tumor cells
wherein the Ras protein is activated as a result of oncogenic
mutation in genes other than the Ras gene.
26. A method of inhibiting farnesyl protein transferase comprising
the administration of an effective amount of the compound of claim
1.
27. A pharmaceutical composition for inhibiting the abnormal growth
of cells comprising an effective amount of compound of claim 1 in
combination with a pharmaceutically acceptable carrier.
28. A method of treating tumor cells expressing an activated ras
oncogene comprising administering an effective amount of a compound
of claim 22.
29. The method of claim 28 wherein the cells treated are pancreatic
tumor cells, lung cancer cells, myeloid leukemia tumor cells,
thyroid follicular tumor cells, myelodysplastic tumor cells,
epidermal carcinoma tumor cells, bladder carcinoma tumor cells,
colon tumors cells, breast tumor cells or prostate tumor cells.
30. A method of treating tumor cells wherein the Ras protein is
activated as a result of oncogenic mutation in genes other than the
Ras gene comprising administering an effective amount of a compound
of claim 1.
31. A method of inhibiting farnesyl protein transferase comprising
the administration of an effective amount of the compound of claim
22.
32. A pharmaceutical composition for inhibiting the abnormal growth
of cells comprising an effective amount of compound of claim 22 in
combination with a pharmaceutically acceptable carrier.
Description
BACKGROUND
[0001] WO 95/10516, published Apr. 20, 1995 discloses tricyclic
compounds useful for inhibiting farnesyl protein transferase.
[0002] In view of the current interest in inhibitors of farnesyl
protein transferase, a welcome contribution to the art would be
compounds useful for the inhibition of farnesyl protein
transferase. Such a contribution is provided by this invention.
SUMMARY OF THE INVENTION
[0003] This invention provides compounds useful for the inhibition
of farnesyl protein transferase (FPT). The compounds of this
invention are represented by the formula: 3
[0004] or a pharmaceutically acceptable salt or solvate thereof,
wherein: one of a, b, c and d represents N or NR.sup.9 wherein
R.sup.9 is O.sup.-, --CH.sub.3 or --(CH.sub.2).sub.nCO.sub.2H
wherein n is 1 to 3, and the remaining a, b, c and d groups
represent CR.sup.1 or CR.sup.2; or
[0005] each of a, b, c, and d are independently selected from
CR.sup.1 or CR.sup.2;
[0006] each R.sup.1 and each R.sup.2 is independently selected from
H, halo, --CF.sub.3, --OR.sup.10 (e.g., --OCH.sub.3), --COR.sup.10,
--SR.sup.10 (e.g., --SCH.sub.3 and --SCH.sub.2C.sub.6H.sub.5),
--S(O).sub.tR.sup.11 (wherein t is 0, 1 or 2, e.g., --SOCH.sub.3
and --SO.sub.2CH.sub.3), --SCN, --N(R.sup.10).sub.2,
--NR.sup.10R.sup.11, --NO.sub.2, --OC(O)R.sup.10,
--CO.sub.2R.sup.10, --OCO.sub.2R.sup.11, --CN, --NHC(O)R.sup.10,
--NHSO.sub.2R.sup.10, --CONHR.sup.10, --CONHCH.sub.2CH.sub.2OH,
--NR.sup.10COOR.sup.11, 4
[0007] --SR.sup.11C(O)OR.sup.11 (e.g.,
--SCH.sub.2CO.sub.2CH.sub.3), --SR.sup.11N(R.sup.75).sub.2 wherein
each R.sup.75 is independently selected from H and --C(O)OR.sup.11
(e.g., --S(CH.sub.2).sub.2NHC(O)O-t-b- utyl and
--S(CH.sub.2).sub.2NH.sub.2), benzotriazol-1-yloxy,
tetrazol-5-ylthio, or substituted tetrazol-5-ylthio (e.g., alkyl
substituted tetrazol-5-ylthio such as 1-methyl-tetrazol-5-ylthio),
alkynyl, alkenyl or alkyl, said alkyl or alkenyl group optionally
being substituted with halo, --OR.sup.10 or --CO.sub.2R.sup.10;
[0008] R.sup.3 and R.sup.4 are the same or different and each
independently represents H, any of the substituents of R.sup.1 and
R.sup.2, or R.sup.3 and R.sup.4 taken together represent a
saturated or unsaturated C.sub.5-C.sub.7 fused ring to the benzene
ring (Ring III);
[0009] R.sup.5, R.sup.6, R.sup.7 and R.sup.8 each independently
represents H, --CF.sub.3, --COR.sup.10, alkyl or aryl, said alkyl
or aryl optionally being substituted with --OR.sup.10, --SR.sup.10,
--S(O).sub.tR .sup.11, --NR.sup.10COOR.sup.11, --N(R.sup.10).sub.2,
--NO.sub.2, --COR.sup.10, --OCOR.sup.10, --OCO.sub.2R.sup.11,
--CO.sub.2R.sup.10, OPO.sub.3R.sup.10), or R.sup.5 is combined with
R.sup.6 to represent .dbd.O or .dbd.S and/or R.sup.7 is combined
with R.sup.8 to represent .dbd.O or .dbd.S;
[0010] R.sup.10 represents H, alky, aryl, or aralkyl (e.g.,
benzyl);
[0011] R.sup.11 represents alkyl or aryl;
[0012] X represents N, CH or C, which C may contain an optional
double bond (represented by the dotted line) to carbon atom 11:
[0013] the dotted line between carbon atoms 5 and 6 represents an
optional double bond, such that when a double bond is present, A
and B independently represent --R.sup.10, halo, --OR .sup.11,
--OCO.sub.2R.sup.11 or --OC(O)R.sup.10, and when no double bond is
present between carbon atoms 5 and 6, A and B each independently
represent H.sub.2, --(OR.sup.11).sub.2; H and halo, dihalo, alkyl
and H, (alkyl).sub.2, --H and --OC(O)R.sup.10, H and --OR.sup.10,
.dbd.O, aryl and H, .dbd.NOR.sup.10 or --O--(CH.sub.2).sub.p--)--
wherein p is 2, 3 or 4; and
[0014] W represents a group selected from the group consisting of:
5
[0015] wherein:
[0016] R.sup.12 is selected from the group consisting of: (a) H;
(b) alkyl; (c) aralkyl (e.g., benzyl): and (d) heteroarylalkyl
(heteroaralkyl) (e.g., --CH.sub.2-imidazolyl);
[0017] R.sup.13 and R.sup.14 are each independently selected from
the group consisting of: (a) H; (b) --C(O)OR.sup.16 wherein
R.sup.16 represents alkyl, aralkyl, and heteroaralkyl; (c)
--SO.sub.2R.sup.17 wherein R.sup.17 is selected from the group
consisting of: --NH.sub.2, --N(alkyl).sub.2 wherein each alkyl is
the same or different (e.g., --N(CH.sub.3).sub.2), alkyl (e.g.,
C.sub.1-6 alkyl, such as methyl), aryl, aralkyl, heteroaryl and
heteroaralkyl; (d) --C(O)R.sup.18 wherein R.sup.18 is selected from
the group consisting of: aryl (e.g., phenyl), alkyl, aralkyl,
heteroaryl, and heteroaralkyl; (e) C.sub.1-6 alkyl; (f) alkaryl;
and (g) C.sub.3-6 cycloalkyl;
[0018] r is 0, 1 or 2;
[0019] s represents 1, 2, 3, 4, or 5 (preferably 3 or 4), and each
Y for each --CY.sub.2-- group is independently selected from H or
--OH, provided that both Y substituents of each --CY.sub.2-- group
are not --OH, and provided that for the --CY.sub.2-- group alpha to
the nitrogen both Y substituents are H, preferably each Y is H such
that each --CY.sub.2-- group is a --CH.sub.2-- group, such that the
group 6
[0020] forms a 3, 4, 5, 6, or 7 (preferably 5 or 6) membered ring
(e.g., piperidyl or pyrrolidinyl),;
[0021] v is 0, 1 or 2;
[0022] R.sup.15 is selected from the group consisting of:
[0023] (a) heteroaryl (e.g., imidazolyl);
[0024] (b) a group selected from: 7
[0025] (5) --CH(OCH.sub.2CH.sub.3).sub.2,
[0026] (6) --OH, and
[0027] (7) --CN; and
[0028] (c) heterocycloalkyl selected from the group consisting of:
8
[0029] z is 0, 1, 2, 3, 4, or 5 wherein each --CH.sub.2-- group is
optionally substituted with a --OH group, i.e., each H of each
--CH.sub.2-- group can optionally be replaced with a --OH group and
the optional substitution on each --CH.sub.2-- group is independent
of the substitution on any other --CH.sub.2-- group, generally each
--CH.sub.2-- is unsubstituted;
[0030] R.sup.22 represents a group selected from: 9
[0031] (5) alkyl (e.g., --CH.sub.3),
[0032] (6) --OR.sup.23 wherein R.sup.23 is selected from the group
consisting of: alky, aryl and H, and 10
[0033] wherein R.sup.24 and R.sup.25 are independently selected
from the group consisting of: --NH.sub.2, alkoxy (e.g.,
--OCH.sub.3), --OH, --CH.sub.2CO.sub.2H, --OCH.sub.2Ph (i.e.,
--OCH.sub.2C.sub.6H.sub.5), --CH(OCH.sub.3)CH(CH.sub.3).sub.2
11
[0034] alkyl, aryl, H, aralkyl, and heteroaralkyl; or R.sup.24 and
R.sup.25 taken ,I together form a carbon chain having 4 or 5
(--CH.sub.2--) groups such that R.sup.24 and R.sup.25 taken
together with the nitrogen to which they are bound form a 5 or 6
membered heterocycloalkyl ring.
[0035] The compounds of this invention: (i) potently inhibit
farnesyl protein transferase, but not geranylgeranyl protein
transferase I, in vitro; (ii) block the phenotypic change induced
by a form of transforming Ras which is a farnesyl acceptor but not
by a form of transforming Ras engineered to be a geranylgeranyl
acceptor; (iii) block intracellular processing of Ras which is a
farnesyl acceptor but not of Ras engineered to be a geranylgeranyl
acceptor; and (iv) block abnormal cell growth in culture induced II
by transforming Ras.
[0036] The compounds of this invention inhibit farnesyl protein
transferase and the farnesylation of the oncogene protein Ras.
[0037] Thus, this invention further provides a method of inhibiting
farnesyl protein transferase, (e.g., ras farnesyl protein
transferase) in mammals, especially humans, by the administration
of an effective amount of the tricyclic compounds described
above.
[0038] The administration of the compounds of this invention to
patients, to inhibit farnesyl protein transferase, is useful in the
treatment of the cancers described below.
[0039] This invention provides a method for inhibiting or treating
the abnormal growth of cells, including transformed cells, by
administering an effective amount of a compound of this invention.
Abnormal growth of cells refers to cell growth independent of
normal regulatory mechanisms (e.g., loss of contact inhibition).
This includes the abnormal growth of: (1) tumor cells (tumors)
expressing an activated Ras oncogene; (2) tumor cells in which the
Ras protein is activated as a result of oncogenic mutation in
another gene; and (3) benign and malignant cells of other
proliferative diseases in which aberrant Ras activation occurs.
[0040] This invention also provides a method for inhibiting or
treating tumor growth by administering an effective amount of the
tricyclic compounds, described herein, to a mammal (e.g., a human)
in need of such treatment. In particular, this invention provides a
method for inhibiting or treating the growth of tumors expressing
an activated Ras oncogene by the administration of an effective
amount of the above described compounds. Examples of tumors which
may be inhibited or treated include, but are not limited to, lung
cancer (e.g., lung adenocarcinoma), pancreatic cancers (e.g.,
pancreatic carcinoma such as, for example, exocrine pancreatic
carcinoma), colon cancers (e.g., colorectal carcinomas, such as,
for example, colon adenocarcinoma and colon adenoma), myeloid
leukemias (for example, acute myelogenous leukemia (AML)), thyroid
follicular cancer, myelodysplastic syndrome (MDS), bladder
carcinoma, epidermal carcinoma, breast cancer and prostate
cancer.
[0041] It is believed that this invention also provides a method
for inhibiting or treating proliferative diseases, both benign and
malignant, wherein Ras proteins are aberrantly activated as a
result of oncogenic mutation in other genes--i.e., the Ras gene
itself is not activated by mutation to an oncogenic form--with said
inhibition or treatment being accomplished by the administration of
an effective amount of the tricyclic compounds described f herein,
to a mammal (e.g., a human) in need of such treatment. For example,
the benign proliferative disorder neurofibromatosis, or tumors in
which Ras is activated due to mutation or overexpression of
tyrosine kinase oncogenes (e.g., neu, src, abli lck, and fyn), may
be inhibited or treated by the tricyclic compounds described
herein.
[0042] The tricyclic compounds useful in the methods of this
invention inhibit or treat the abnormal growth of cells. Without
wishing to be bound by theory, it is believed that these compounds
may function through the inhibition of G-protein function, such as
ras p21, by blocking G-protein isoprenylation, thus making them
useful in the treatment of proliferative diseases such as tumor
growth and cancer. Without wishing to be bound by theory, it is
believed that these compounds inhibit ras farnesyl protein
transferase, and thus show antiproliferative activity against ras
transformed cells.
DETAILED DESCRIPTION OF THE INVENTION
[0043] As used herein, the following terms are used as defined
below unless otherwise indicated:
[0044] Ac--represents acetyl;
[0045] MH.sup.+--represents the molecular ion plus hydrogen of the
molecule in the mass spectrum;
[0046] M.sup.+--represents the molecular ion of the molecule in the
mass spectrum;
[0047] benzotriazol-1-yloxy represents 12
[0048] 1-methyl-tetrazol-5-ylthio represents 13
[0049] alkenyl-represents straight and branched carbon chains
having at least one carbon to carbon double bond and containing
from 2 to 12 carbon atoms, preferably from 2 to 6 carbon atoms and
most preferably from 3 to 6 carbon atoms;
[0050] alkynyl-represents straight and branched carbon chains
having at least one carbon to carbon triple bond and containing
from 2 to 12 carbon atoms, preferably from 2 to 6 carbon atoms;
[0051] alkyl-(including the alkyl portions of aralkyl and
heteroarylalkyl)-represents straight and branched carbon chains and
contains from one to twenty carbon atoms, preferably one to six
carbon atoms;
[0052] aralkyl-represents an aryl group, as defined below, bound to
an alkyl group, as defined above, preferably the alkyl group is
--CH.sub.2--, (e.g., benzyl);
[0053] aryl (including the aryl portion of aralkyl and
aralkyl)-represents a carbocyclic group containing from 6 to 15
carbon atoms and having at least one aromatic ring (e.g., aryl is a
phenyl ring), with all available substitutable carbon atoms of the
carbocyclic group being intended as possible points of attachment,
said carbocyclic group being optionally substituted (e.g., 1 to 3)
with one or more of halo, alkyl, hydroxy, alkoxy, phenoxy,
CF.sub.3, amino, alkylamino, dialkylamino, --COOR.sup.10 or
--NO.sub.2;
[0054] BOC-represents --C(O)OC(CH.sub.3).sub.3;
[0055] --CH.sub.2-imidazolyl represents an imidazolyl group bound
by any substitutable carbon of the imidazole ring to a
--CH.sub.2--, that is: 14
[0056] such as --CH.sub.2-(2-, 4- or 5-)imidazolyl, for example
15
[0057] Et-represents ethyl;
[0058] halo-represents fluoro, chloro, bromo and iodo;
[0059] heteroaryl-represents cyclic groups, optionally substituted
with R.sup.3, R.sup.4, phenyl, and or --CH.sub.2C(O)OCH.sub.3, said
cyclic groups having at least one heteroatom selected from 0, S or
N, said heteroatom interrupting a carbocyclic ring structure and
having a sufficient number of delocalized pi electrons to provide
aromatic character, with the aromatic heterocyclic groups
preferably containing from 2 to 14 carbon atoms, e.g., (2-, 4- or
5-)imidazolyl, triazolyl, 16
[0060] 2-, 3- or 4-pyridyl or pyridyl N-oxide (optionally
substituted with R.sup.3 and R.sup.4), wherein pyridyl N-oxide can
be represented as: 17
[0061] heteroarylalkyl (heteroaralkyl)-represents a heteroaryl
group, as defined above, bound to an alkyl group, as defined above,
preferably the alkyl group is --CH.sub.2-- (e.g., --CH.sub.2-(4- or
5-)imidazolyl);
[0062] heterocycloalkyl-represents a saturated, branched or
unbranched carbocylic ring containing from 3 to 15 carbon atoms,
preferably from 4 to 6 carbon atoms, which carbocyclic ring is
interrupted by 1 to 3 hetero groups selected from --O--, --S-- or
--NR.sup.10--, suitable heterocycloalkyl groups include: (1) 2- or
3- tetrahydrofuranyl, (2) 2- or 3- tetrahydrothienyl, (3) 2-, 3- or
4- piperidinyl, (4) 2- or 3-pyrrolidinyl, (5) 2- or 3-piperizinyl,
and (6) 2- or 4-dioxanyl; and
[0063] Ph-represents phenyl
[0064] The following solvents and reagents are referred to herein
by the abbreviations indicated: tetrahydrofuran (THF);
[0065] isopropanol (iPrOH); ethanol (EtOH); methanol (MeOH); acetic
acid (HOAc or AcOH); ethyl acetate (EtOAc); N,N-dimethyl-formamide
(DMF); trifluoroacetic acid (TFA); trifluoro-acetic anhydride
(TFAA); 1-hydroxybenzotriazole (HOBT);
1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (DEC);
diisobutylaluminum hydride(DIBAL); and 4-methylmorpholine
(NMM).
[0066] Reference to the position of the substituents R.sup.1,
R.sup.2, R.sup.3, and R.sup.4 is based on the numbered ring
structure: 18
[0067] Those skilled in the art will also appreciate that the S and
R stereochemistry at the C-11 bond are: 19
[0068] Compounds of Formula 1.0 include compounds wherein the
bottom piperidinyl group is a 4- or 3-piperidinyl group, i.e.,
20
[0069] Compounds of Formula 1.0 include compounds wherein R.sup.2
and R.sup.4 are H, and R.sup.1 and R.sup.3 are halo (preferably
independently selected from Br or Cl). For example, R.sup.1 is Br
and R.sup.3 is Cl. These compounds include compounds wherein
R.sup.1 is in the 3-position and R.sup.3 is in the 8-position,
e.g., 3-Br and 8-Cl. Compounds of Formula 1.0 also include
compounds wherein R.sup.2 is H, and R.sup.1, R.sup.3 and R.sup.4
are halo (preferably independently selected from Br or Cl).
[0070] Preferably, compounds of Formula 1.0 are represented by
compounds of Formula 1.1: 21
[0071] wherein all substituents are as defined for Formula 1.0.
[0072] Preferably, R.sup.2 is H and R.sup.1, R.sup.3 and R.sup.4
are halo; a is N and b, c and d are carbon; A and B are each
H.sub.2; the optional bond between C5 and C6 is absent; X is CH;
and R.sup.5, R.sup.6, R.sup.7 and R.sup.8 are H. More preferably,
R.sup.1, R.sup.3 and R.sup.4 are independently selected from Br or
Cl. Most preferably, R.sup.1 is Br, and R.sup.3 and R.sup.4 are
independently selected from Cl and Br.
[0073] More preferably, compounds of Formula 1.0 are represented by
compounds of Formula 1.2 and Formula 1.3: 22
[0074] and most preferably, compounds of Formulas 1.4 and 1.5
23
[0075] wherein R.sup.1, R.sup.3 and R.sup.4 are each independently
selected from halo, preferably, Br or Cl; and A, B, X and W are as
defined for Formula 1.0. More preferably, A and B are each H.sub.2;
the optional bond between C5 and C6 is absent; and X is CH. Most
preferably, R.sup.1 is Br; R.sup.3 and R.sup.4 are independently Br
or Cl, and still more preferably R.sup.3 is Cl and R.sup.4 is Br; A
and B are each H.sub.2; the optional t bond between C5 and C6 is
absent; X is CH; and R.sup.5, R.sup.6, R.sup.7 and R.sup.8 are
H.
[0076] Examples of R.sup.15 include: 24
[0077] When W represents: 25
[0078] and r is 0: (1) preferably, R.sup.12 is selected from the
group consisting of: (a) H; (b) alkyl; (c) aralkyl; and (d)
heteroaralkyl; and most preferably, R.sup.12 is selected from the
group consisting of: (a) H, (b) methyl, (c) --CH.sub.2-imidazolyl
and (d) benzyl; (2) preferably, R.sup.13 and R.sup.14 are
independently selected from the group consisting of: (a) H; (b)
--C(O)OR.sup.16 wherein R.sup.16 is alkyl; (c) --SO.sub.2R.sup.17
wherein R.sup.17 is alkyl or aryl; (d) --C(O)R.sup.18 wherein
R.sup.18 is aryl; and (e) alky; and most preferably, R.sup.13 and
R.sup.14 are independently selected from the group consisting of:
(a) H, (b) --C(O)OC(CH.sub.3).sub.3, (c) --SO.sub.2CH.sub.3 and (d)
--C(O)-phenyl. Preferably, when one of R.sup.13 or R.sup.14 is
--C(O)OR.sup.16, --SO.sub.2R.sup.17, --C(O)R.sup.18, alkaryl or
cycloalkyl, the remaining R.sup.13 or R.sup.14 is H. Preferred
combinations of substituent groups include: (1) R.sup.12 being
alkylw (more preferred, methyl), R.sup.13 being --C(O)OR.sup.16
(more preferred --C(O)OC(CH.sub.3).sub.3) and R.sup.14 being H; (2)
R.sup.12 being heteroarylalkyl (more preferred --CH.sub.2-(4 or
5)-imidazolyl), R.sup.13 being H or --C(O)OR.sup.16 (more preferred
H or --C(O)OC(CH.sub.3).sub.3) and R.sup.14 being H; (3) R.sup.12
being aralkyl (more preferred benzyl), R.sup.13 being
--C(O)OR.sup.16 (more preferred --C(O)OC(CH.sub.3).sub.3) and
R.sup.14 being H; (4) R.sup.12 being H, R.sup.13 being
--C(O)OR.sup.16 (more preferred --C(O)OC(CH.sub.3).sub.3) and
R.sup.14 being H: (5) R.sup.12 being H, R.sup.13 being
--SO.sub.2R.sup.17 (more preferred --SO.sub.2CH.sub.3) and R.sup.14
being H; and (6) R.sup.12 being H, R.sup.13 being --C(O)R.sup.18
(more preferred --C(O)-phenyl) and l R.sup.14 being H.
[0079] Those skilled in the art will appreciate that the W
substituent described in the previous paragraph can be derived from
known amino acids having one carboxyl and amino group. F Examples
of such amino acids include but are not limited to glycine,
alanine, phenylalanine, asparagine and histidine. For example, see
Morrison and Boyd, Organic Chemistry, Fifth Edition, Allyn and
Bacon, Inc., Boston, pages 1346-1347, the disclosure of which is
incorporated herein by reference thereto.
[0080] When W represents 26
[0081] and r is 1 or 2, preferably R.sup.12 is H, and R.sup.13 and
R.sup.14 are independently selected from alkyl, most preferably,
R.sup.13 and R.sup.14 are the same alkyl group (e.g., methyl).
[0082] When W represents 27
[0083] s is preferably 3, such that a pyrrolidone ring is formed,
and R.sup.13 is preferably H or --C(O)OR.sup.16 wherein R.sup.16 is
alkyl; most preferably, R.sup.13 is H or
--C(O)OC(CH.sub.3).sub.3.
[0084] When W represents: 28
[0085] and v is 0, preferably R.sup.12 represents H and R.sup.15
represents heteroaryl or heterocycloalkyl. Most preferably, when
R.sup.15 is heteroaryl said heteroaryl is imidazolyl 29
[0086] and when R.sup.15 is heterocycloalkyl said heterocycloalkyl
is 30
[0087] When W represents: 31
[0088] and v is 1 or 2, preferably R.sup.12 represents H and
R.sup.15 is heterocycloalkyl. Most preferably R.sup.12 represents H
and R.sup.15 is heterocycloalkyl, e.g., 32
[0089] When W represents 33
[0090] and z is 0, preferably R.sup.22 represents 34
[0091] and R.sup.24 and R.sup.25 preferably represent H.
[0092] When W represents 35
[0093] and z is 1, 2, 3, 4 or 5, R.sup.22 preferably represents
--OR.sup.23 and R.sup.23 preferably represents alkyl and most
preferably methyl.
[0094] Compounds of Formulas 1.2A and 1.3A: 36
[0095] are preferred when X is CH or N, and R.sup.1, R.sup.3 and
R.sup.4 are halo.
[0096] The preferred compounds of this invention are represented by
the compounds of Formulas: 37
[0097] wherein R.sup.1, R.sup.3 and R.sup.4 are halo and the
remaining substituents are as defined above, with the compounds of
Formula 1.5A being more preferred.
[0098] Those skilled in the art will appreciate that W substituent:
38
[0099] wherein r is 0 includes 39
[0100] and W substituent: 40
[0101] wherein v is 0 includes 41
[0102] Representative compounds of Formula 1.0 wherein W is 42
[0103] and r is 0 include: 43 44
[0104] and r is 1 or 2 include: 45
[0105] Representative compounds of Formula 1.0 wherein W is 46
[0106] and s is 3 include: 47
[0107] Representative compounds of Formula 1.0 wherein W is 48
[0108] and v is 0 include: 49
[0109] Representative compounds of Formula 1.0 wherein W is 50
[0110] and v is 1 include: 51
[0111] Compounds of Formula 1.0 wherein W is 52
[0112] and z is 0 include: 53
[0113] Compounds of Formiula 1.0 wherein W is 54
[0114] and z is 1, 2, 3, 4 or 5 include: 55
[0115] Compounds of this invention also include: 56
[0116] or pharmaceutically acceptable salts or solvates
thereof.
[0117] The compounds of this invention also include the
1--N-oxides--i.e, for example, compounds of the the formula: 57
[0118] wherein represents the remainder of the compound, or
pharmaceutically acceptable salts or solvates thereof.
[0119] Optical rotation of the compounds ((+)- or (-)-) are
measured in methanol or ethanol at 25.degree. C.
[0120] This invention includes the above compounds in the amorphous
state or in the cyrstalline state.
[0121] Lines draw,n into the ring systems indicate that the
indicated bond may be attached to any of the substitutable ring
carbon atoms.
[0122] Certain compounds of the present invention may exist in
different isomeric forms (e.g., enantiomers or diastereoisomers)
including atropisomers (i.e., compounds wherein the 7-membered ring
is in a fixed conformation such that the 11-carbon atom is
positioned above or below the plane of the fused benzene rings due
to the presence of a 10-bromo substituent). The invention
contemplates all such isomers both in pure form and in admixture,
including racemic mixtures. Enol forms are also included.
[0123] Certain tricyclic compounds will be acidic in nature, e.g.
those compounds which possess a carboxyl or phenolic hydroxyl
group. These compounds may form pharmaceutically acceptable salts.
Examples of such salts may include sodium, potassium, calcium,
aluminum, gold, silver and lithium salts. For example, compounds
having the --OR.sup.23 group wherein R.sup.23 is H can form a
sodium or lithium salt--i.e., a compound with a --ONa or --OLi
group. Also contemplated are salts formed with pharmaceutically
acceptable amines such as ammonia, alkyl amines,
hydroxyalkylamines, N-methylglucamine and the like.
[0124] Certain basic tricyclic compounds also form pharmaceutically
acceptable salts, e.g., acid addition salts. For example, the
pyrido-nitrogen atoms may form salts with strong acid, while
compounds having basic substituents such as amino groups also form
salts with weaker acids. Examples of suitable acids for salt
formation are hydrochloric, sulfuric, phosphoric, acetic, citric,
oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic,
maleic, methanesulfonic and other mineral and carboxylic acids well
known to those in the art. The salts are prepared by contacting the
free base form with a sufficient amount of the desired acid to
produce a salt in the conventional manner. The free base forms may
be regenerated by treating the salt with a suitable dilute aqueous
base solution such as dilute aqueous NaOH, potassium carbonate,
ammonia and sodium bicarbonate. The free base forms differ from
their respective salt forms somewhat in certain physical
properties, such as solubility in polar solvents, but the acid and
base salts are otherwise equivalent to their respective free base
forms for purposes of the invention.
[0125] All such acid and base salts are intended to be
pharmaceutically acceptable salts within the scope of the invention
and all acid and base salts are considered equivalent to the free
forms of the corresponding compounds for purposes of the
invention.
[0126] Compounds of the invention may be prepared according to the
procedures described in WO 95/10516 published Apr.20, 1995,
Application Serial No. 08/410,187 filed Mar. 24, 1995, Application
Serial No. 08/577,951 filed Dec. 22, 1995 (now abandoned),
Application Serial No. 08/615,760 filed Mar. 13, 1996 (now
abandoned), WO 97/23478 published Jul. 3, 1997 which discloses the
subject matter of Serial No. 08/577,951 and 08/615,760, Application
Serial No. 08/710,225 filed Sep. 13, 1996, and Application Serial
No. 08/877,453 filed Jun. 17, 1997; the disclosures of each being
incorporated herein by reference thereto; and according to the
procedures described below.
[0127] Compounds of the invention can be prepared by reacting a
compound of the formula: 58
[0128] wherein all substituents are as defined for Formula 1.0,
with the appropriate protected piperidinyl acetic acid (e.g.,
1-N-t-butoxy-carbonylpiperidinyl acetic acid together with
DEC/HOBT/NMM in DMF at about 25.degree. C. for about 18 hours to
produce a compound of the formula: 59
[0129] The compound of Formula 21.0 is then reacted either with TFA
or 10% sulfuric acid in dioxane and methanol followed by NaOH to
produce the compound of Formula 20.0 60
[0130] For example, the compound of formula 61
[0131] can be prepared by reaction of a compound of Formula 19.0
with 1-N-t-butoxy-carbonylpiperidinyl-4-acetic acid as described
above.
[0132] For example, compounds of Formula 22.0 include the
compounds: 62
[0133] The preparation of these compounds are described in
Preparative Examples 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 and 13,
respectively, below.
[0134] The compounds of the invention can be prepared by reacting a
compound of the formula: 63
[0135] with the appropriate protected piperidinyl acetic acid
(e.g., 1-N-t-butoxycarbonylpiperidinyl acetic acid together with
DEC/HOBT/NMM in DMF at about 25.degree. C. for about 18 hours to
produce a compound of the formula: 64
[0136] The compound of Formula 21.1 is then reacted either with TFA
or 10% sulfuric acid in dioxane and methanol followed by NaOH to
produce the compound of Formula 22.1 65
[0137] The amide compounds of this invention, represented by
Formula 1.7 66
[0138] can be prepared by reacting the compound of Formula 22.1
with the appropriate carboxylic acid in the presence of a coupling
agent such as DEC and HOBT in dimethylformamide. Alternatively, the
compound of Formula 22.1 can be reacted with an acid chloride or
anhydride in a solvent such as pyridine.
[0139] The W group on Formula 1.7 can contain functionality that
can be converted to other functionality by methods, such as
hydrolysis, that are well known in the art. For example, the
compound of Formula 16.0-B, can be converted to the compound of
Formula 74-B, and the compound of Formula 35.0-B to the compound of
Formula 52.0-B by treatment with methanolic potassium hydroxide
followed by acid. Also, compounds of Formulas 86.0-B and 89.0-B can
be converted to compounds of Formulas 88.0-B and 90.0-B,
respectively, by treatment with acids such as trifluoroacetic acid
or dioxane saturated with HCl gas.
[0140] Compounds having an 1-N--O group: 67
[0141] can be prepared from the corresponding pyridyl compounds:
68
[0142] by oxidation with meta-chloroperoxybenzoic acid. This
reaction is conducted in a suitable organic solvent, e.g.,
dichloromethane (usually anhydrous) or methylene chloride, at a
suitable temperature, to produce the compounds of the invention
having the N--O substituent at position 1 of Ring I of the
tricyclic ring system.
[0143] Generally, the organic solvent solution of the starting
tricyclic reactant is cooled to about 0C before the
m-chloroper-oxybenzoic acid is added. The reaction is then allowed
to warm to room temperature during the reaction period. The desired
product can be recovered by standard separation means. For example,
the reaction mixture can be washed with an aqueous solution of a
suitable base, e.g., saturated sodium bicarbonate or NaOH (e.g., 1N
NaOH), and then dried over anhydrous magnesium sulfate. The
solution containing the product can be concentrated in vacuo. The
product can be purified by standard means, e.g., by chromatography
using silica gel (e.g., flash column chromatography).
[0144] Alternatively, N--O compounds can be made from intermediate:
69
[0145] by the above oxidation procedure with m-chloroperoxybenzoic
acid and 70
[0146] wherein is a protecting group, e.g., BOC. After oxidation
the protecting group is removed by techniques well knowrn in the
art. The N--O intermediate is then reacted further to produce the
compounds of the invention.
[0147] Compounds of Formula 19.0 include the compound of Formula
19.1: 71
[0148] The compound of Formula 19.1 is prepared by methods known in
the art, for example by methods disclosed in WO 95/10516, in U.S.
Pat. No. 5,151,423 and those described below. The above
intermediate compound can also b e prepared by a procedure
comprising the following steps:
[0149] (a) reacting an amide of the formula 72
[0150] wherein R.sup.11a is Br, R.sup.5a is hydrogen and R.sup.6a
is C.sub.1-C.sub.6 alkyl, aryl or heteroaryl; R.sup.5a is
C.sub.1-C.sub.6 alkyl, aryl or heteroaryl and R.sup.6a is hydrogen;
R.sup.5a and R.sup.6a are independently selected from the group
consisting of C.sub.1-C.sub.6 alkyl and aiyl; or R.sup.5a and
R.sup.6a, together with the nitrogen to which they are attached,
form a ring comprising 4 to 6 carbon atoms or comprising 3 to 5
carbon atoms and one hetero moiety selected from the group
consisting of --O-- and --NR.sup.9a_, wherein R.sup.9a is H,
C.sub.1-C.sub.6 alkyl or phenyl;
[0151] with a compound of the formula 73
[0152] wherein R.sup.1a, R.sup.2a, R.sup.3a and R.sup.4a are
independently selected from the group consisting of hydrogen and
halo and R.sup.7a is Cl or Br, in the presence of a strong base to
obtain a compound of the formula 74
[0153] (b) reacting a compound of step (a) with
[0154] (i) POCl.sub.3 to obtain a cyano compound of the formula
75
[0155] (ii) DIBALH to obtain an aldehyde of the formula 76
[0156] (c) reacting the cyano compound or the aldehyde with a
piperidine derivative of the formula 77
[0157] wherein L is a leaving group selected from the group
consisting of Cl and Br, to obtain a ketone or an alcohol of the
formula below, respectively: 78
[0158] (d)(i) cyclizing the ketone with CF.sub.3SO.sub.3H to obtain
a compound of Formula 13.0a wherein the dotted line represents a
double bond; or
[0159] (d)(ii) cyclizing the alcohol with polyphosphoric acid to
obtain an Intermediate compound wherein the dotted line represents
a single bond.
[0160] Methods for preparing the Intermediate compounds disclosed
in WO 95/10516, U.S. Pat. No. 5,151,423 and described below employ
a tricyclic ketone intermediate. Such intermediates of the formula
79
[0161] wherein R.sup.11b, R.sup.1a, R.sup.2a, R.sup.3a and R.sup.4a
are independently selected from the group consisting of hydrogen
and halo, can be prepared by the following process comprising:
[0162] (a) reacting a compound of the formula 80
[0163] (i) with an amine of the formula NHR.sup.5aR.sup.6a, wherein
R.sup.5a and R.sup.6a are as defined in the process above; in the
presence of a palladium catalyst and carbon monoxide to obtain an
amide of the formula: 81
[0164] (ii) with an alcohol of the formula R.sup.10aOH, wherein
R.sup.10a is C.sub.1-C.sub.6 lower alkyl or C.sub.3-C.sub.6
cycloalkyl, in the presence of a palladium catalyst and carbon
monoxide to obtain the ester of the formula 82
[0165] followed by reacting the ester with an amine of formula
NHR.sup.5aR.sup.6a to obtain the amide;
[0166] (b) reacting the amide with an iodo-substituted benzyl
compound of the formula 83
[0167] wherein R.sup.1a, R.sup.2a, R.sup.3a, R.sup.4a and R.sup.7a
are as defined above, in the presence of a strong base to obtain a
compound of the formula 84
[0168] (c) cycliziing a compound of step (b,) with a reagent of the
formula R.sup.8aMgL, wherein R.sup.8a is C.sub.1-C.sub.8 alkyl,
aryl or heteroaiyl and L is Br or Cl, provided that prior to
cyclization, compounds wherein R.sup.5a or R.sup.6a is hydrogen are
reacted with a suitable N-protecting group.
[0169] (+)-Isomers of compounds of Formula 19.2 85
[0170] can be prepared with high enantioselectivity by using a
process comprising enzyme catalyzed transesterification.
Preferably, a racemic compound of Formula 19.3 86
[0171] is reacted with an enzyme such as Toyobo LIP-300 and an
acylating agent such as trifluoroetihly isobutyrate; the resultant
(+)-amide is then isolated from the (-)-enantiomeric amine by
techniques well known in the art, and then the (+)-amide is
hydrolyzed, for example by refluxing with an acid such as
H.sub.2SO.sub.4, and the resulting compound is then reduced with
DIBAL by techniques well known in the art to obtain the
corresponding optically enriched (+)-isomer of Formula 19.2.
Alternatively, a racemic compound of Formula 19.3, is first reduced
to the corresponding racemic compound of Formula 19.2 and then
treated with the enzyme (Toyobo LIP-300) and acylating agent as
described above to obtain the (+)-amide, which is hydrolyzed to
obtain the optically enriched (+)-isomer.
[0172] Those skilled in the art will appreciate that compounds of
Formula 1.0 having other R.sup.1, R.sup.2, R.sup.3 and R.sup.4
substituents may be made by the above enzyme process.
[0173] To produce the compounds of Formula 1.0, wherein W is 87
[0174] r is 0, and R.sup.13 and R.sup.14 are selected from H or
--C(O)OR.sup.16, the compounds of Formulas 20.0 or 22.0 are reacted
with the appropriate protected amino acid: 88
[0175] in the presence of DEC and HOBt in dimethylformamide to
produce a compound of the formula: 89
[0176] respectively.
[0177] Reaction of compounds of Formulas 23.0 or 24.0 with TFA in
methylene chloride results in the deprotected compounds: 90
[0178] respectively.
[0179] Compounds of Formula 1.0 wherein W is 91
[0180] r is 0, R.sup.12 is H, R.sup.13 or R.sup.14 is H, and the
remaining R.sup.13 or R.sup.14 is --C(O)OR.sup.16 can be prepared
by reacting a compound of Formula 1.0 wherein W is 92
[0181] r is 0, R.sup.12 is H, and R.sup.13 and R.sup.14 are both H,
with the appropriate chloroformate 93
[0182] TEA and CH.sub.2Cl.sub.2.
[0183] Compounds of Formulas 25.0 or 26.0 wherein R.sup.13 is
selected from --SO.sub.2R.sup.17 or --C(O)R.sup.18 can be prepared
by reacting a compound of Formula 25.0 or 26.0 with a suitable
sulfonyl chloride (R.sup.17SO.sub.2Cl) or a suitable acyl chloride
(R.sup.18C(O)Cl) with TEA in a suitable organic solvent i(e.g.,
CH.sub.2Cl.sub.2).
[0184] Compounds of Formula 1.0 wherein W is 94
[0185] r is 1 or 2 and R.sup.12 is H can be prepared by reacting a
compound of Formula 20.0 or 22.0 with the appropriately substitued
carboxylic acid and, for example DEC, HOBT and N-methylmorpholine,
or by reacting a compound of Formula 20.0 or 22.0 with the
appropriately substituted acid chloride.
[0186] For example, a compound of Formula 20.0 or 22.0 can be
reacted with 95
[0187] from propionic acid, wherein R.sup.13 and R.sup.14 are, for
example, allyl (e.g., methyl). Where the amino carboxylic acid is
not commercially available, it can be prepared by reaction of ethyl
acrylate with the appropriate amino compound (as described by Ahn,
K. H. et al., Tetrahedron Letters, 35, 1875-1878 (1994)) with
subsequent hydrolysis of the ester to the desired aminocarboxylic
acid.
[0188] Also, for example, a compound of Formula 20.0 or 22.0 can be
reacted with 96
[0189] from butyric acid, wherein R.sup.13 and R.sup.14 are, for
example, alkyl (e.g., methyl). Where the amino c~arboxylic acid is
not commercially available, the appropriate acid chloride can be
prepared in a manner similar to that described by Goel, O. P. et
al., Synthesis, p. 538 (1973). The acid chloride is then reacted
with a compound of Formula 20.0 or 22.0 to give the compound 97
[0190] respectively. The chloro atom can be then displaced with the
appropriate amine to give the desired compound.
[0191] Where either R.sup.13 or R.sup.14 is H, then the starting
material would be a protected amino carbcixylic acid 98
[0192] wherein Z is an appropriater protecting group (e.g., BOC,
CBZ (carbonylbenzyloxy) or TFA). Coupling this protected amino
carboxylic acid with a compound of Formula 20.0 or 22.0 would then
give the amino protected intermediate 99
[0193] respectively. The amino protected intermediate (20.OB or
22.OB) would then be alkylated, and then the protecting group
removed, using standard procedures known in the art.
[0194] Compounds of Formula 1.0 wvherein W is 100
[0195] v is 0, and R.sup.12 is H can be prepared by reacting a
compound of Formula 20.0 or 22.0 with chloroacetylchloride, TEA and
CH.sub.2Cl.sub.2 to produce a compound of the formula: 101
[0196] The chloro atom in the --C(O)CH.sub.2Cl group in the
compound of Formula 26.0 or 27.0 is then displaced with an
appropriate nucleophile, R.sup.15, using a suitable base, e.g.,
sodium carbonate, and optionally, a suitable suitable solvent
(e.g., DMF).
[0197] Compounds of Formula 1.0 wherein W is 102
[0198] z is 0, and R.sup.22 is 103
[0199] can be prepared from compounds of Formula 20.0 or 22.0 by
reaction with oxallyl chloride and an excess of the amine 104
[0200] Compounds of Formula 1.0 wherein W is 105
[0201] z is 1, 2, 3, 4 or 5 and R.sup.22 is --OR.sup.23, and
R.sup.23 is, for example, alkyl, can be prepared by reaction of a
compound of Formula 20.0 or 22.0 with the appropriate substituted
dicarboxylic acid which is protected as a mono ester with an
appropriate alkyl or airyl group. The corresponding acids (i.e.,
R.sup.23 is H) can be obtained by base hydrolysis (e.g., NaOH) of
the ester. The compounds, wherein R.sup.22 is --NR.sup.24R.sup.25,
can be prepared by reacting the appropriately substituted amine
with the carbo,yhlic acid generated above using DEC, HOBT and NMM.
For example, for compounds wherein z is 3 a glutarate 106
[0202] (wherein R.sup.23 is alky, e.g., methyl) can be used, and
for compounds wherein z is 2 a succinate 107
[0203] (wherein R.sup.23 is alkyl, e.g., methyl) can be used, and
for compounds wherein z is 1 a malonate 108
[0204] (wherein R.sup.23 is alkyl, e.g., ethyl) can be used.
[0205] Reaction Scheme 1 illustrates the preparation of compounds
of this invention. 109
[0206] Compounds useful in this invention are exemplified by the
following examples, which should not be construed to limit the
scope of the disclosure.
PREPARATIVE EXAMPLE 1
[0207] 110
[0208] Combine 10 g (60.5 mmol) of ethyl 4-pyridylacetate and 120
mL of dry CH.sub.2Cl.sub.2 at -20.degree. C., add 10.45 g (60.5
mmol) of MCPBA and stir at -20.degree. C. for 1 hour and then at
25.degree. C. for 67 hours. Add an additional 3.48 g (20.2 minoles)
of MCPBA and stir at 25.degree. C. for 24 hours. Dilute with
CH.sub.2Cl.sub.2 and wash with saturated NaHCO.sub.3 (aqueous) and
then water. Diy over MgSO.sub.4, concentrate in vacuo to a residue,
and chromatograph (silica gel, 2%-5.5% (10% NH.sub.4OH in
MeOH)/CH.sub.2Cl.sub.2) to give 8.12 g of the product compound.
Mass Spec.: MH.sup.+=182.15 111
[0209] Combine 3.5 g (19.3 mmol) of the product of Step A, 17.5 mL
of EtOH and 96.6 mL of 10% NaOH (aqueous) and heat the mixture at
67.degree. C. for 2 hours. Add 2 N HCl (aqueous) to adjust to
pH=2.37 and concentrate in vacuio to a residue. Add 200 mL of dry
EtOH, filter through celite.RTM. and wash the filter cake with dry
EtOH (2.times.50 ml). Concentrate the combined filtrates in vacuo
to give 2.43 g of the title compound. 112
PREPARATIVE EXAMPLE 2
[0210] The title compound is prepared via the process disclosed in
PCT International Publication No. WO95/10516.
PREPARATIVE EXAMPLE 3
[0211] 113
[0212] Combine 14.95 g (39 nmmol) of
8-chloro-11-(1-ethoxy-carbonyl-4-pipe-
ridinyl)-11H-benzo[5,6]cyclohepta[1,2-b]pyridine and 150 mL of
CH.sub.2Cl.sub.2, then add 13.07 g (42.9 nmmol) of
(nBu).sub.4NNO.sub.3 and cool the mixture to 0.degree. C. Slowly
add (dropwise) a solution of 6.09 mL (42.9 mmol) of TFAA in 20 mL
of CH.sub.2Cl.sub.2 over 1.5 hours. Keep the mixture at 0.degree.
C. overnight, then wash successively with saturated NaHCO.sub.3
(aqueous), water and brine. Dry the organic solution over
Na.sub.2SO.sub.4, concentrate in vacuo to a residue and
chromatograph the residue (silica gel, EtOAc/hexane gradient) to
give 4.32 g and 1.90 g of the two product compounds 3A(i) and
3A(ii), respectively.
[0213] Mass Spec. for compound 3A(i): MH.sup.+=428.2;
[0214] Mass Spec. for compound 3A(ii): MH+=428.3. 114
[0215] Combine 22.0 g (51.4 mnmol) of the product 3A(i) from Step
A, 150 mL of 85% EtOH (aqueous), 25.85 g (0.463 mole) of Fe powder
and 2.42 g (21.8 mmol) of CaCl.sub.2, and heat at reflux overnight.
Add 12.4 g (0.222 mole) of Fe powder and 1.2 g (10.8 mmol) of
CaCl.sub.2 and heat at reflux for 2 hours. Add another 12.4 g
(0.222 mole) of Fe powder and 1.2 g (10.8 mmol) of CaCl.sub.2 and
heat at reflux for 2 hours more. Filter the hot mixture through
celite.RTM., wash the celite.RTM. with 50 mL of hot EtOH and
concentrate the filtrate in vacuo to a residue. Add 100 mL of
anhydrous EtOH, concentrate to a residue and chromatograph the
residue (silica gel, MeOH/CH.sub.2Cl.sub.2 gradient) to give 16.47
g of the product compound. 115
[0216] Combine 16.47 g (41.4 mmol) of the product from Step B, and
150 mL of 48% HBr (aqueous) and cool to -3.degree. C. Slowly add
(dropwise) 18 mL of bromine, then slowly add (dropwise) a solution
of 8.55 g (0.124 mole) of NaNO.sub.2 in 85 mL of water. Stir for 45
minutes at -3.degree. to 0.degree. C., then adjust to pH =10 by
adding 50% NaOH (aqueous). Extract with EtOAc, wash the extracts
with brine and dry the extracts over Na.sub.2SO.sub.4. Concentrate
to a residue and chromatograph (sflica gel, EtOAc/hexane gradient)
to give 10.6 g and 3.28 g of the two product compounds 3C(i) and
3C(ii), respectively.
[0217] Mass Spec. for compound 3,C(i): MH.sup.+=461.2;
[0218] Mass Spec. for compound 3,C(ii): MH.sup.+=539. 116
[0219] Hydrolyze the product 3C(i) of Step C by dissolving in
concentrated HCl and heating to about 100.degree. C. for @ 16
hours. Cool the mixture, the neutralize with 1 M NaOH (aqueous).
Extract with CH.sub.2Cl.sub.2, dry the extracts over MgSO.sub.4,
filter and concentrate in vacuo to the title compound.
[0220] Mass Spec.: MH.sup.+=466.9. 117
[0221] Dissolve 1.160 g (2.98 mmol) of the title compound from Step
D in 20 mL of DMF, stir at room temperature, and add 0.3914 g (3.87
nmuol) of 4-methyl-morpholine, 0.7418 g (3.87 mmol) of DEC, 0.5229
g (3.87 mmol) of HOBT, and 0.8795 g (3.87 mmol) of
1-N-t-butoxycarbonyl-piperidinyl-4-acet- ic acid. Stir the mixture
at room temperature for 2 days, then concentrate in vacuo to s
residue and partition the residue between CH.sub.2Cl.sub.2 and
water. Wash the organic phase successively with saturated
NaHCO.sub.3 (aqueous), 10% NaH.sub.2PO.sub.4 (aqueous) and brine.
Dry the organic phase over MgSO.sub.4, filter and concentrate in
vacuo to a residue. Chromatograph the residue (silica gel, 2%
MeOH/CH.sub.2Cl.sub.2+NH.sub.3) to give 1.72 g of the product.
m.p.=94.0-94.5.degree. C., Mass Spec.: MH.sup.+=616.3,
[0222] elemental analysis:
[0223] calculated--C, 60.54; H, 6.06; N, 6.83
[0224] found--C, 59.93; H, 6.62; N, 7.45. 118
[0225] Combine 1.67 g (2.7 mmol) of the product of Step E and 20 mL
of CH.sub.2Cl.sub.2 and stir at 0.degree. C. Add 20 mL of TFA, stir
the mixture for 2 hours, then basify the mixture with 1 N NaOH
(aqueous). Extract with CH.sub.2Cl.sub.2, dry the organic phase
over MgSO.sub.4, filter and concentrate in vacuo to give 1.16 g of
the product. m.p.=140.2-140.8.degree. C., Mass Spec.:
MH.sup.+=516.2.
[0226] PREPARATIVE EXAMPLE 4 119
[0227] Combine 25.86 g (55.9 mmol) of
4-(8-chloro-3-bromo-5,6-dihydro-11H--
benzo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1
-piperidine-1-carboxylic acid ethyl ester and 250 mL of
concentrated H.sub.2SO.sub.4 at -5.degree. C., then add 4.8 g (56.4
mmol) of NaNO.sub.3 and stir for 2 hours. Poui the mixture into 600
g of ice and basify with concentrated NH.sub.40H (aqueous). Filter
the mixture, wash with 300 nL of water, then extract with 500 mL of
CH.sub.2Cl.sub.2. Wash the extract with 200 mL of water, dry over
MgSO.sub.4, then filter and concentrate in vacuo to a residue.
Chromatograph the residue (silica gel, 10% EtOAc/CH.sub.2Cl.sub.2)
to give 24.4 g (86% yield) of the product. m.p.=165-167.degree. C.,
Mass Spec.: MH.sup.+=506 (CI),
[0228] elemental analysis:
[0229] calculated--C, 52.13; H, 4.17; N, 8.29
[0230] found--C, 52.18; H, 4.51; N, 8.16. 120
[0231] Combine 20 g (40.5 mmol) of the product of Step A and 200 mL
of concentrated H.sub.2SO.sub.4 at 20.degree. C., then cool the
mixture to 0.degree. C. Add 7.12 g (24.89 mmol) of
1,3-dibromo-5,5-dimethyl-hydantoi- n to the mixture and stir for 3
hours at 20.degree. C. Cool to 0.degree. C., add an additional 1.0
g (3.5 mmol) of the dibromohydantoin and stir at 20.degree. C. for
2 hours. Pour the mixture into 400 g of ice, basify with
concentrated NH.sub.4OH (aqueous) at 0.degree. C., and collect the
resulting solid by filtration. Wash the solid with 300 mL of water,
slurry in 200 mL of acetone and filter to provide 19.79 g (85.6%
yield) of the product. m.p.=236-237.degree. C., Mass Spec.:
MH.sup.+=584 (CI),
[0232] elemental analysis:
[0233] calculated--C, 45.11; H, 3.44; N, 7.17
[0234] found--C, 44.95; H, 3.57; N, 7.16. 121
[0235] Combine 25 g (447 mmol) of Fe filings, 10 g (90 mmol) of
CaCl.sub.2 and a suspension of 20 g (34.19 mmol) of the product of
Step B in 700 mL of 90:10 EtOH/water at 50.degree. C. Heat the
mixture at reflux overnight, filter through Celite.RTM. and wash
the filter cake with 2.times.200 mL of hot EtOH. Combine the
filtrate and washes, and concentrate in vacuo to a residue. Extract
the residue with 600 mL of CH.sub.2Cl.sub.2, wash with 300 mL of
water and dry over MgSO.sub.4. Filter and concentrate in vacuo to a
residue, then chromatograph (silica gel, 30%
EtOAc/CH.sub.2Cl.sub.2) to give 11.4 g (60% yield) of the product.
m.p.=211-212.degree. C., Mass Spec.: MH.sup.+=554 (CI),
[0236] elemental analysis:
[0237] calculated--C, 47.55; H, 3.99; N, 7.56
[0238] found--C, 47.45; H, 4.31; N, 7.49. 122
[0239] Slowly add (in portions) 20 g (35.9 mmol) of the product of
Step C to a solution of 8 g (116 mmol) of NaNO.sub.2 in 120 mL of
concentrated HCl (aqueous) at -10.degree. C. Stir the resulting
mixture at 0.degree. C. for 2 hours, then slowly add (dropwise) 150
mL (1.44 mole) of 50% H.sub.3PO.sub.2 at 0.degree. C. over a 1 hour
period. Stir at 0.degree. C. for 3 hours, then pour into 600 g of
ice and basify with concentrated NH.sub.4OH (aqueous). Extract with
2.times.300 mL of CH.sub.2Cl.sub.2, dry the extracts over
MgSO.sub.4, then filter and concentrate in vacuo to a residue.
Chromatograph the residue (silica gel, 25% EtOAc/hexanes) to give
13.67 g (70% yield) of the product. m.p.=163-165.degree. C., Mass
Spec.: MH.sup.+=539 (CI),
[0240] elemental analysis:
[0241] calculated--C, 48.97; H, 4.05; N, 5.22
[0242] found--C, 48.86; H, 3.91; N, 5.18. 123
[0243] Combine 6.8 g (12.59 mmol) of the product of Step D and 100
mL of concentrated HCl (aqueous) and stir at 85.degree. C.
overnight. Cool the mixture, pour it into 300 g of ice and basify
with concentrated NH.sub.4OH (aqueous). Extract with 2.times.300 mL
of CH.sub.2Cl.sub.2, then dry the extracts over MgSO.sub.4. Filter,
concentrate in vacuo to a residue, then chromatograph (silica gel,
10% MeOH/EtOAc+2% NH.sub.4OH (aqueous)) to give 5.4 g (92% yield)
of the title compound. m.p.=172-174.degree. C., Mass Spec.:
MH.sup.+=467 (FAB),
[0244] elemental analysis:
[0245] calculated--C, 48.69; H, 3.65; N, 5.97
[0246] found--C, 48.83; H, 3.80; N, 5.97.
[0247] Step F:
[0248] Following essentially the same procedure as Step C of
Preparative Example 5 below, the title compound from Step E above
is reacted with 1-N-t-butoxycarbonylpiperidinyl-4-acetic acid to
produce the compound 124
[0249] Step G:
[0250] Following essentially the same procedure as Step D of
Preparative Example 5 below, the title compound from Step F above
is deprotected to yield the title compound of Preparative Example
4.
PREPARATIVE EXAMPLE 5
[0251] 125
[0252] Hydrolyze 2.42 g of
4-(8-cliloro-3-bromo-5,6-dihydro-11H-benzo[5,6]-
cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylic acid
ethyl ester via substantially the same procedure as described in
Preparative Example 3, Step D, to give 1.39 g (69% yield) of the
product. 126
[0253] Combine 1 g (2.48 mmol) of the product of Step A and 25 mL
of dry toluene, add 2.5 mL of 1 M DIBAL in toluene and heat the
mixture at reflux. After 0.5 hours, add another 2.5 mL of 1 M DIBAL
in toluene and heat at reflulx for 1 hour. (The reaction is
monitored by TLC using 50% MeOH/CH.sub.2Cl.sub.2+NH.sub.4OH
(aqueous).) Cool the mixture to room temperature, add 50 mL of 1 N
HCl (aqueous) and stir for 5 min. Add 100 mL of 1 N NaOH (aqueous),
then extract with EtOAc (3.times.150 mL). Dry the extracts over
MgSO.sub.4, filter and concentrate in vacuo to give 1.1 g of the
title compound. 127
[0254] Combine 0.501 g (1.28 mmol) of the title compound of Step B
and 20 mL of dry DMF, then acid 0.405 g (1.664 mmol) of
1--N-t-butoxycarbonylpip- eridinyl-4-acetic acid, 0.319 g (1.664
mmol) of DEC, 0.225 g (1.664 mmol) of HOBT, and 0.168 g (1.664
mmol) of 4-methylmorpholine and stir the mixture at room
temperature overnight. Concentrate the mixture in vacuo to a
residue, then partition the residue between 150 mL of
CH.sub.2Cl.sub.2 and 150 mL of saturated NaHCO.sub.3 (aqueous).
Extract the aqueous phase with another 150 mL of CH.sub.2Cl.sub.2.
Dry the organic phase over MgSO.sub.4, and concentrate in lacuo to
a residue. Chromatograph the residue (silica gel, 500 mL hexane, 1
L of 1% MeOH/CH.sub.2Cl.sub.2+0.1% NH.sub.4OH (aqueous), then 1 L
of 2% MeOH/CH.sub.2Cl.sub.2+0.1% NH.sub.4OH (aqueous)) to give
0.575 g of the product. m.p.=115.degree.-125.degree. C.; Mass
Spec.: MH.sup.+=616. 128
[0255] Combine 0.555 g (0.9 mmol) of the product of Step C and 15
mL of CH.sub.2Cl.sub.2 and cool the mixture to 0.degree. C. Add 15
mL of TFA and stir at 0.degree. C. for 2 hours. Concentrate in
vacuo at 40-45.degree. C. to a residue, then partition the residue
between 150 mL of CH.sub.2Cl.sub.2 and 100 mL of saturated
NaHCO.sub.3 (aqueous). Extract the aqueous layer with 100 mL of
CH.sub.2Cl.sub.2, comnbine the extracts and dry over MgSO.sub.4.
Concentrate in vacuo to give 0.47 g of the product.
[0256] m.p.=140.degree.-150.degree. C; Mass Spec.:
MH.sup.+=516.
PREPARATIVE EXAMPLE 6
[0257] 129
[0258] [racemic as well as (+-and (-)-isomers] 130
[0259] Combine 16.6 g (0.03 mole) of the product of Preparative
Example 4, Step D, with a 3:1 solution of CH.sub.3CN and water
(212.65 mL CH.sub.3CN and 70.8 mL of water) and stir the resulting
slurry overnight at room temperature. Add 32.833 g (0.153 mole) of
NaIO.sub.4 and then 0.31 g (2.30 mmol) of RuO.sub.2 and stir at
room temperature give 1.39 g (69% yield) of the product. (The
addition of RuO is accompanied by an exothermic reaction and the
temperature climbs from 20.degree. to 30.degree. C.) Stir the
mixture for 1.3 hrs. (temperature returned to 25.degree. C. after
about 30 min.), then filter to remove the solids and wash the
solids with CH.sub.2Cl.sub.2. Concentrate the filtrate in vacuo to
a residue and dissolve the residue in CH.sub.2Cl.sub.2. Filter to
remove insoluble solids and wash the solids with CH.sub.2Cl.sub.2.
Wash the filtrate with water, concentrate to a volume of about 200
mL and wash with bleach, then with water. Extract with 6 N HCl
(aqueous). Cool the aqueous extract to 0.degree. C. and slowly add
50% NaOH (aqueous) to adjust to pH=4 while keeping the temperature
<30.degree. C. Extract twice with CH.sub.2Cl.sub.2, dry over
MgSO.sub.4 and concentrate in vCLcuo to a residue. Slurny the
residue in 20 mL of EtOH and cool to 0.degree. C. Collect the
resulting solids by filtration and dry the solids in vacuo to give
7.95 g of the product. .sup.1H NMR (CDCl.sub.3, 200 MHz): 8.7 (s,
1H); 7.85 (m, 6H); 7.5 (d, 2H); 3.45 (m, 2H); 3.15 (m, 2H). 131
[0260] Combine 21.58 g (53.75 mmol) of the product of Step A and
500 mL of an anhydrous 1:1 mixture of EtOH and toluene, add 1.43 g
(37.8 mmol) of NaBH.sub.4 and heat the mixture at reflux for 10
min. Cool the mixture to 0.degree. C., add 100 mL of water, then
adjust to pH=4-5 with 1 M HCl (aqueous) while keeping the
temperature <10.degree. C. Add 250 mL of EtOAc and separate the
layers. Wash the organic layer with brine (3.times.50 mL) then dry
over Na2SO.sub.4. Concentrate in vacuo to a residue (24.01 g) and
chromatograph the residue (silica gel, 30 %
hexane/CH.sub.2Cl.sub.2) to give the product. Impure fractions were
purified by rechromatography. A total of 18.57 g of the product was
obtained. .sup.1H NMR (DMSO-d.sub.6, 400 MHz): 8.5 (s, 1H); 7.9 (s,
1H); 7.5 (d of d, 2H); 6.2 (s, 1H); 6.1 (s, 1H); 3.5 (m, 1H); 3.4
(m, 1H); 3.2 (m, 2H). 132
[0261] Combine 18.57 g (46.02 mmol) of the product of Step B and
500 mL of CHCl.sub.3, then add 6.70 mL (91.2 mmol) of SOCl.sub.2,
and stir the mixture at room temperature for 4 hrs. Add a solution
of 35.6 g (0.413 mole) of piperazine in 800 mL of THF over a period
of 5 min. and stir the mixture for 1 hr. at room temperature. Heat
the mixture at reflux overnight, then cool to room temperature and
dilute the mixture with 1 L of CH.sub.2Cl.sub.2. Wash with water
(5.times.200 mL), and extract the aqueous wash with CHCl.sub.3
(3.times.100 mL). Combine all of the organic solutions, wash with
brine (3.times.200 mL) and cdry over MgSO.sub.4. Concentrate in
vacuo to a residue and chromatograph (silica gel, gradient of 5%,
7.5%, 10% MeOH/CH.sub.2Cl.sub.2+NH.sub.4OH) to give 18.49 g of the
title compound as a racemic mixture. 133
[0262] The racemic title compouncl of Step C is separated by
preparative chiral chromatography (Chiralpack AD, 5 cm.times.50 cm
column, flow rate 100 mL/min., 20% iPrOH/hexane+0.2% diethylamine),
to give 9.14 g of the (+)-isomer and 9.30 g of the (-)-isomer.
[0263] Physical chemical data for (+)-isomer:
m.p.=74.5.degree.-77.5.degre- e. C.; Mass Spec. MH.sup.+=471.9;
[.alpha.].sub.D.sup.25=+97.4.degree. (8.48 mg/2mL MeOH).
[0264] Physical chemical data for (-)-isomer:
m.p.=82.9.degree.-84.5.degre- e. C.; Mass Spec. MH.sup.+=471.8;
[.alpha.].sub.D.sup.25=-97.4.degree. (8.32 mg/2mL MeOH). 134
[0265] Combine 3.21 g (6.80 mmol) of the (-)-isomer product of Step
D and 150 mL of anhydrous DMF. Add 2.15 g (8.8 mmol) of
1-N-t-butoxycarbonylpip- eridinyl-4-acetic acid, 1.69 g (8.8 mmol)
of DEC, 1.19 g (8.8 mmol) of HOBT and 0.97 mL (8.8 mmol) of
N-methylmorpholine and stir the mixture at room temperature
overnight. Concentrate in vacuo to remove the DMF and add 50 mL of
saturated NaHCO.sub.3 (aqueous). Extract with CH.sub.2Cl.sub.2
(2.times.250 mL), wash the extracts with 50 mL, of brine and dry
over MgSO.sub.4. Concentrate in vacuo to a residue and
chromatograph (silica gel, 2% MeOH/CH.sub.2Cl.sub.2+10% NH.sub.4OH)
to give 4.75 g of the product. m.p.=75.7.degree.-78.5.degree. C.;
Mass Spec.: MH.sup.+=697; [.alpha.].sub.D.sup.25=5.50 (6.6 mg/2 mL
MeOH). 135
[0266] Combine 4.70 g (6.74 mmol) of the product of Step E and 30
mL of MeOH, then add 50 mL of 10% H.sub.2SO.sub.4/dioxane in 10 mL
aliquots over a 1 hr. period. Flour the mixture into 50 mL of water
and add 15 mL of 50% NaOH (aqueous) to adjust to pH.apprxeq.10-11.
Filter to remove the resulting solids and extract the filtrate with
CH.sub.2Cl.sub.2 (2.times.250 mL). Concentrate the aqueous layer in
vacuo to remove the MeOH and extract again with 250 mL of
CH.sub.2Cl.sub.2. Dry the combined extracts over MgSO.sub.4 and
concentrate in vacuo to give the product.
m.p.=128.1.degree.-131.5.degree. C.; Mass Spec.: MH.sup.+=597;
[.alpha.].sub.D.sup.25=6.02.degree. (9.3 mg/2 mL MeOH).
PREPARATIVE EXAMPLE 7
[0267] 136
[0268] Combine 15 g (38.5 mmol) of
4-(8-chloro-3-bromo-5,6-dihydro-11H-ben-
zo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylic
acid ethyl ester and 150 mL of concentrated H.sub.2SO.sub.4 at
-5.degree. C., then add 3.89 g (38.5 mmol) of KNO.sub.3 and stir
for 4 hours. Pour the mixure into 3 L of ice and basify with 50%
NaOH (aqueous). Extract with CH.sub.2Cl.sub.2, dry over MgSO.sub.4,
then filter and concentrate in vacuo to a residue. Recrystallize
the residue from acetone to give 6.69 g of the product. .sup.1H NMR
(CDCl.sub.3, 200 MHz): 8.5 (s, 1H); 7.75 (s, 1H); 7.6 (s, 1H); 7.35
(s, 1H); 4.15 (q, 2H); 3.8 (m, 2H); 3.5-3.1 (m, 4H); 3.0-2.8 (m,
2H); 2.6-2.2 (m, 4H); 1.25 (t, 3H). 137
[0269] Combine 6.69 g (13.1 mmol) of the product of Step A and 100
mL of 85% EtOH/water, then add 0.66 g (5.9 mmol) of CaCl.sub.2 and
6.56 g (117.9 mmol) of Fe and heat the mixture at reflux overnight.
Filter the hot reaction mixture through celite.RTM. and rinse the
filter cake with hot EtOH. Concentrate the filtrate in vacuo to
give 7.72 g of the product. Mass Spec.: MH.sup.+=478.0. 138
[0270] Combine 7.70 g of the product of Step B and 35 mL of HOAc,
then add 45 mL of a solution of Br.sub.2 in HOAc and stir the
mixture at room temperature overnight. Add 300 mL of 1 N NaOH
(aqueous), then 75 mL of 50% NaOH (aqueous) and extract with EtOAc.
Dry the extract over MgSO.sub.4 and concentrate in vacuo to a
residue. Chromatograph the residue (silica gel, 20%-30%
EtOAc/hexane) to give 3.47 g of the product (along with another
1.28 g of partially purified product). Mass Spec.:
MH.sup.+=555.9.
[0271] .sup.1H NMR (CDCl.sub.3, 300 MHz): 8.5 (is, 1H); 7.5 (s,
1H); 7.15 (s, 1H); 4.5 (s, 2H); 4.15 (m, 3H); 3.8 (br s, 2H);
3.4-3.1 (m, 4H); 9-2.75 (m, 1H); 2.7-2.5 (m, 2H); 2.4-2.2 (m, 2H);
1.25 (m, 3H). 139
[0272] Combine 0.557 g (5.4 mmol of t-butylnitrite and 3 mL of DMF,
and heat the mixture at to 60.degree.-70.degree. C. Slowly add
(dropwise) a mixture of 2.00 g (3.6 mmol) of the product of Step C
and 4 mL of DMF, then cool the mixture to room temperature. Add
another 0.64 mL of t-butylnitrite at 40.degree. C. and reheat the
mixture to 60.degree.-70.degree. C. for 0.5 hrs. Cool to room
temperature and pour the mixture into 150 mL of water. Extract with
CH.sub.2Cl.sub.2, dry the extract over MgSO.sub.4 and concentrate
in vacuo to a residue. Chromatograph the residue (silica gel,
10%-20% EtOAc/hexane) to give 0.74 g of the product. Mass Spec.:
MH.sup.+=541.0.
[0273] .sup.1H NMR (CDCl3, 200 MHz): 8.52 (s, 1H); 7.5 (d, 2H); 7.2
(s, 1H); 4.15 (q, 2H); 3.9-3.7 (m, 2H): 3.5-3.1 (m, 4H); 3.0-2.5
(m, 2H); 2.4-2.2 (m, 2H); 2.1-1.9 (m, 2H); 1.26 (t, 3H). 140
[0274] Combine 0.70 g (1.4 mmol) of the product of Step D and 8 mL
of concentrated HCl (aqueous) and heat the mixture at reflux
overnight. Add 30 mL of 1 N NaOH (aqueous), then 5 mL of 50% NaOH
(aqueous) and extract with CH.sub.2Cl.sub.2. Dry the extract over
MgSO.sub.4 and concentrate in vacuo to give 0.59 g of the title
compound. Mass Spec.: M.sup.+=468.7.
m.p.=123.9.degree.-124.2.degree. C. 141
[0275] React 6.0 g (12.8 mmnol) of the title compound from Step E
and with 3.78 g (16.6 mmol) of
1-N-t-butoxycarbonylpiperidinyl-4-acetic acid using substantially
the same procedures as described for Preparative Example 5, Step C,
to give 8.52 g of the product. Mass Spec.: MH.sup.+=694.0 (FAB).
.sup.1H NMR (CDCl.sub.3, 200 MHz): 8.5 (d, 1H); 7.5 (d, 2H); 7.2
(d, 1H); 4.15-3.9 (m, 3H); 3.8-3.6 (m, 1H); 3.5-3.15 (m, 3H); 2.9
(d, 2H); 2.8-2.5 (m, 4H); 2.4-1.8 (m, 6H); 1.8-1.6 (br d, 2H); 1.4
(s, 9H); 1.25-1.0 (m, 2H). 142
[0276] Combine 8.50 g of the product of Step F and 60 mL of
CH.sub.2Cl.sub.2, then cool to 0.degree. C. and add 55 mL of TFA.
Stir the mixture for 3 h at 0.degree. C., then add 500 mL of 1 N
NaOH (aqueous) followed by 30 mL of 50% NaOH (aqueous). Extract
with CH.sub.2Cl.sub.2, dry over MgSO.sub.4 and concentrate in vacuo
to give 7.86 g of the product. Mass Spec.: M+=593.9 (FAB). .sup.1H
NMR (CDCl.sub.3, 200 MHz): 8.51 (d, 1H); 7.52 (d of d, 2H); 7.20
[d, 1H); 4.1-3.95 (m, 2H); 3.8-3.65 (m, 2H); 3.5-3.05 (m, 5H):
3.0-2.5 (m, 6H); 2.45-1.6 (m, 6H); 1.4-1.1 (m, 2H).
PREPARATIVE EXAMPLE 8
[0277] 143
[0278] [racemic as well as (+)- and (-)-isomers] 144
[0279] Prepare a solution of 8.1 g of the title compound from
Preparative Example 7, Step E, in toluene and add 17.3 mL of a 1M
solution of DIBAL in toluene. Heat the mixture at reflux and slowly
add (dropwise) another 21 mL of 1 M DIBAL/toluene solution over a
period of 40 min. Cool the reaction mixture to about 0.degree. C.
and add 700 mL of 1 M HCl (aqueous). Separate and discard the
organic phase. Wash the aqueous phase with CH.sub.2Cl.sub.2,
discard the extract, then basify the aqueous phase by adding 50%
NaOH (aqueous). Extract with CH.sub.12Cl.sub.2, dry the extract
over MgSO.sub.4 and concentrate in vacuo to give 7.30 g of the
title compound, which is a racemic mixture of enantiomers. 145
[0280] The racemic title compound of Step A is separated by
preparative chiral chromatography (Chiralpack AD, 5 cm.times.50 cm
column, using 20% iPrOH/hexane+0.2% diethylamine), to give the
(+)-isomer and the (-)-isomer of the title compound.
[0281] Physical chemical data for (+)-isomer: m.p.=148.8.degree.
C.; Mass Spec. MH.sup.+=469; [.alpha.].sub.D.sup.25=+65.60 (12.93
mg/2 mL MeOH).
[0282] Physical chemical data for (-)-isomer: m.p.=112.degree. C.;
Mass Spec. MH.sup.+=469; [.alpha.].sub.D.sup.25=-65.2.degree. (3.65
mg/2 mL MeOH). 146
[0283] React 1.33 g of the (+)-isomer of the title compound of
Preparative Example 8, Step B, with 1.37 g of
1-N-t-butoxy-carbonylpiperidinyl-4-acet- ic acid using
substantially the same procedures as described for Preparative
Example 5, Step C, to give 2.78 g of the product. Mass Spec.:
MH.sup.+=694.0 (FAB); [.alpha.].sub.D.sup.25=+34.1.degree. (5.45
mg/2 mL, MeOH). 147
[0284] Treat 2.78 g of the product of Step C via substantially the
same procedure as described for Preparative Example 5, Step D, to
give 1.72 g of the product. m.p.=104.1.degree. C.; Mass Spec.:
MH.sup.+=594; [.alpha.].sub.D.sup.25=+53.40 (11.42 mg/2 mL,
MeOH).
PREPARATIVE EXAMPLE 9
[0285] 148
[0286] [racemic as well as (+)- and (-)-isomers] 149
[0287] Combine 40.0 g (0.124 mole) of the starting ketone and 200
mL of H.sub.2SO.sub.4 and cool to 0.degree. C. Slowly add 13.78 g
(0. 136 mole) of KNO.sub.3 over a period of .1.5 hrs., then warm to
room temperature and stir overnight. Work up the reaction using
substantially the same procedure as described for Preparative
Example 4, Step A. Chromatograph (silica gel, 20%, 30%, 40%, 50%
EtOAc/hexane, then 100% EtOAc) to give 28 g of the 9-nitro product,
along with a smaller quantity of the 7-nitro product and 19 g of a
mixture of the 7-nitro and 9-nitro compounds. 150
[0288] React 28 g (76.2 mmol) of the 9-nitro product of Step A, 400
mL of 85% EtOH/water, 3.8 g (34.3 mmol) of CaCl.sub.2 and 38.28 g
(0.685 mole) of Fe using substantially the same procedure as
described for Preparative Example 4, Step C, to give 24 g of the
product 151
[0289] Combine 13 g (38.5 mmol) of the product of Step B. 140 mL of
HOAC and slowly add a solution of 2.95 mL (57.8 mmol) of Br.sub.2
in 10 mL of HOAc over a period of 20 min. Stir the reaction mixture
at room temperature, then concentrate in vacuo to a residue. Add
CH.sub.2Cl.sub.2 and water, then adjust to pH=8-9 with 50% NaOH
(aqueous). Wash the organic phase with water, then brine and dry
over Na.sub.2SO.sub.4. Concentrate in vacuo to give 11.3 g of the
product. 152
[0290] Cool 100 mL of concentrated HCl (aqueous) to 0.degree. C.,
then add 5.61 g (81.4 mmol) of NaNO.sub.2 and stir for 10 min.
Slowly add (in portions) 11.3 g (27.1 mmol) of the product of Step
C and stir the mixture at 0.degree.-3.degree. C. for 2.25 hrs.
Slowly add (dropwise) 180 mL of 50% H.sub.3PO.sub.2 (aqueous) and
allow the mixture to stand at 0.degree. C. overnight. Slowly add
(dropwise) 150 mL of 50% NaOH over 30 min., to adjust to pH=9, then
extract with CH.sub.2Cl.sub.2. Wash the extract with water, then
brine and dry over Na.sub.2SO.sub.4. Concentrate in vacuo to a
residue and chromatlograph (silica gel, 2% EtOAc/CH.sub.2Cl.sub.2)
to give 8.6 g of the product. 153
[0291] Combine 8.6 g (21.4 mmol) of the product of Step D and 300
mL of MeOH and cool to 0.degree.-2.degree. C. Add 1.21 g (32.1
mmol) of NaBH.sub.4 and stir the mixture at .about.0.degree. C. for
1 hr. Add another 0.121 g (3.21 mmol) of NaBH.sub.4, stir for 2 hr.
at 0.degree. C., then let stand overnight at 0.degree. C.
Concentrate in vacuo to a residue then partition the residue
between CH.sub.2Cl.sub.2 and water. Separate the organic phase and
concentrate in vacuo (50.degree. C.) to give 8.2 g of the product.
154
[0292] Combine 8.2 g (20.3 nmuol) of the product of Step E and 160
mL of CH.sub.2Cl.sub.2, cool to 0.degree. C., then slowly add
(dropwise) 14.8 mL (203 mmol) of SOCl.sub.2 over a 30 min. period.
Warm the mixture to room temperature and stir for 4.5 hrs., then
concentrate in vacuo to a residue, add CH.sub.2Cl.sub.2 and wash
with 1 N NaOH (aqueous) then brine and dry over Na.sub.2SO.sub.4.
Concentrate in vacuo to a residue, then add dry THF and 8.7 g (101
mmol) of piperazine and stir at room temperature overnight.
Concentrate in vacuo to a residue, add CH.sub.2Cl.sub.2, and wash
with 0.25 N NaOH (aqueous), water, then brine. Dry over
Na.sub.2SO.sub.4 and concentrate in vacuo to give 9.46 g of the
crude product. Chromatograph (silica gel, 5%
MeOH/CH.sub.2Cl.sub.2+NH.sub- .3) to give 3.59 g of the title
compound, as a racemate. .sup.1H NMR (CDCl.sub.3, 200 MHz): 8.43
(d, 1H); 7.55 (d, 1H); 7.45 (d, 1H); 7.11 (d, 1H); 5.31 (s, 1H);
4.86-4.65 (m, 1H); 3.57-3.40 (m, 1H); 2.98-2.55 (m, 6H); 2.45-2.20
(m, 5H). 155
[0293] The racemic title compound from Step F (5.7 g) is
chromatographed as described for Preparative Example 6, Step D,
using 30% iPrOH/hexane+0.2% diethylamine, to give 2.88 g of the
R-(+)-isomer and 2.77 g of the S-(-)-isomer of the title
compound.
[0294] Physical chemical data for the R-(+)-isomer: Mass Spec.
MH.sup.+=470.0; [.alpha.].sub.D.sup.25=+12.1.degree. (10.9 mg/2mL
MeOH).
[0295] Physical chemical data for the S-(-)-isomer: Mass Spec.
MH.sup.+=470.0; [.alpha.].sub.D.sup.25=-13.2.degree. (11.51 mg/2mL
MeOH).
[0296] Step H:
[0297] Following essentially the same procedure as Preparative
Example 5, Steps C and D, the racemic title compound of Preparative
Example 9 is obtained from the racemic compound of Step F.
Similarly, using the (-)- or (+)- isomer from Step G, the (-)- or
(+)-isomer of the title compound of Preparative Example 9 is
obtained, respectively.
PREPARATIVE EXAMPLE 10
[0298] 156
[0299] [racemic as well as (+)- and (-)-isomers] 157
[0300] Combine 13 g (33.3 mmol) of the title compound from
Preparative Example 4, Step E, and 300 mL of toluene at 20.degree.
C., then add 32.5 mL (32.5 imuol) of a 1 M solution of DIBAL in
toluene. Heat the mixture at reflux for 1 hr., cool to 20.degree.
C., add another 32.5 mL of 1 M DIBAL solution and heat at reflux
for 1 hr. Cool the mixture to 20.degree. C. and pour it into a
mixture of 400 g of ice, 500 mL of EtOAc and 300 mL, of 10% NaOH
(aqueous). Extract the aqueous layer with CH.sub.2Cl.sub.2
(3.times.200 mL), dry the organic layers over MgSO.sub.4, then
concentrate in vacuo to a residue. Chromatograph (silica gel, 12%
MeOH/CH.sub.2Cl.sub.2+4% NH.sub.4OH) to give 10.4 g of the title
compound as a racemate. Mass Spec.: MH.sup.+=469 (FAB). partial
.sup.1H NMR (CDCl.sub.3, 400 MHz): 8.38 (s, 1H); 7.57 (s, 1H); 7.27
(d, 1H); 7.06 (d, 1H); 3.95 (d, 1H). 158
[0301] The racemic title compound of Step A is separated by
preparative chiral chromatography (Chiralpack AD, 5.times.50 cm
column, using 5% iPrOH/hexane+0.2% diethylamine), to give the
(+)-isomer and the (-)-isomer of the title compound.
[0302] Physical chemical data for (+)-isomer: Mass Spec.
MH.sup.+=469 (FAB); [.alpha.].sub.D.sup.25=+43.5.degree. (c=0.402,
EtOH); partial 1H NMR (CDCl.sub.3, 400 MHz): 8.38 (s, 1H); 7.57 (s,
1H); 7.27 (d, 1H); 7.05 (d, 1H); 3.95 (d, 1H).
[0303] Physical chemical data for (-)-isomer: Mass Spec.
MH.sup.+=469 (FAB); [.alpha.].sub.D.sup.25=-41.8.degree. (c=0.328
EtOH); partial .sup.1H NMR (CDCl.sub.3, 400 MHz): 8.38 (s, 1H);
7.57 (s, 1H); 7.27 (d, 1H); 7.05 (d, 1H); 3.95 (d, 1 H).
[0304] Step C:
[0305] Following the procedure of Preparative Example 9, Step H,
the racemic compound, the (+)-isomer or the (-)-isomer of the title
compound of Preparative Example 10 can be obtained.
PREPARATIVE EXAMPLE 11
[0306] 159
[0307] [racemic as well as R-(+)- and S-(-)-isomers]
[0308] The compound 160
[0309] is prepared according to the procedures of Preparative
Example 40 of WO 95/10516 (published Apr. 20, 1995), by following
the procedures described in Example 1.93 of WO 95/10516.
[0310] The (+)- and (-)-isomers can be separated by following
essentially the same procedure as Step D of Preparative Example
6.
[0311] Physical chemical data for the R-(+)-isomer: .sup.13C NMR
(CDCl.sub.3): 155.8 (C); 146.4 (CH); 140.5 (CH); 140.2 (C); 136.2
(C); 135.3 (C); 133.4 (C); 132.0 (CH); 129.9 (CH); 125.6 (CH);
119.3 (C); 79.1 (CH); 52.3 (CH.sub.2); 52.3 (CH); 45.6 (CH.sub.2);
45.6 (CH.sub.2); 30.0 (CH.sub.2); 29.8 (CH.sub.2).
[.alpha.].sub.D.sup.25=+25.8.degree. (8.46 mg/2 mL MeOH).
[0312] Physical chemical data for the S-(-)-isomer: .sup.13C NMR
(CDCl.sub.3): 155.9 (C); 146.4 (CH); 140.5 (CH); 140.2 (C); 136.2
(C); 135.3 (C); 133.3 (C); 132.0 (CH); 129.9 (CH); 125.5 (CH);
119.2 (C); 79.1 (CH); 52.5 (CH.sub.2); 52.5 (CH); 45.7 (CH.sub.2);
45.7 (CH.sub.2); 30.0 (CH.sub.2); 29.8 (CH.sub.2).
[.alpha.].sub.D.sup.25=-27.9.degree. (8.90 mg/2 mL MeOH).
[0313] Following essentially the same procedure as Preparative
Example 5, Steps C and D, the racemic compound, (+)-isomer or
(-)-isomer of the title compound of Preparative Example 11 can be
obtained from the corresponding racemic compound, (+)-isomer or
(-)-isomer of the compound 161
PREPARATIVE EXAMPLE 12
[0314] 162
[0315] Follow a procedure outlined in Collect. Czech. Chem. Comm.
(1990) 55, 2086. Dissolve 0.2 g 10.915 mmol) of (aminooxy)acetic
acid hemihydrochloride and 0.2 g (3 mmol) of acetone in 2 mL of
pyridine and allow to stand for 18 hr. Concentrate under vacuum and
partition the residue between ethyl acetate and 1 N Hcl. Dry the
organic layer over magnesium sulfate and concentrate under vacuum
to give a white solid mp=77.3-78.degree. C.
PREPARATIVE EXAMPLE 13
[0316] 163
[0317] Follow the procedure of Preparative Example 12 but use
2-aminooxypropionic acid hemihydrochloride instead of
(aminooxy)acetic acid to obtain the product as a colorless oil.
PREPARATIVE EXAMPLE 14
[0318] 164
[0319] Follow the procedure of Preparative Example 12 but use
4-pyridinecarboxaldehyde N-oxide instead of acetone to obtain the
product that was recrystallized from water to give a white solid
mp=227-228.degree. C.
PREPARATIVE EXAMPLE 15
[0320] 165
[0321] Follow the procedure of Preparative Example 12 but use
2-hydroxybenzaldehyde instead of acetone to obtain the product as a
white solid mp=152-153.5.degree. C.
EXAMPLE 1
[0322] 166
[0323] The compound of Formula 28.0 167
[0324] (Preparative Example 8) (0.149 g, 0.25 mmol) was combined
with 1-hydroxybenzotriazole hydrate (0.067 g, 0.5 mmol),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.096
g, 0.5 mmol), N-BOC-glycine (0.087 g, 0.5 mmol) and anhydrous
dimethylformamide (5 mL) and the resulting mixture was stirred at
room temperature under nitrogen overnight. Concentration in vacuo
provided an oil which was diluted with dichloromethane, washed with
1M hydrochloric acid and 1 M aqueous sodium hydroxide, then dried
over anhydrous magnesium sulfate.
[0325] Filtration and concentration in vacuo afforded the compound
of Formula 2.0 (+-isomer) (0.16 g, 85%, mp 116-123.degree. C.).
EXAMPLE 2
[0326] 168
[0327] To the compound of Formula 2.0 (Example 1) (0.145 g)
dissolved in anhydrous dichloromethane (10 nmL) was added
trifluoroacetic acid (2 mL) and the resulting solution was stirred
at room temperature for 1 hour. 50% Aqueous sodium hydroxide was
added slowly followed by dictiloromethane and brine. The mixture
was shaken well, the organic phase was separated and dried over
anhydrous magnesium sulfate. Filtration and concentration in vacuo
afforded the compound of Formula 16.0 (+-isomer) (0.086 g, 68%, mp
131-138.degree. C.).
EXAMPLE 3
[0328] 169
[0329] The compound of Formula 28.0 (Preparative Example 8) (0.10
g, 0.17 mmol) was combined with 1-hydroxybenzotriazole hydrate
(0.045 g, 0.34 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (0.064 g, 0.34 mmol), N-tert-butoxycarbonyl-L-alanine
(0.064 g, 0.34 mmol) and anhydrous dimethylformamide (10 mL) and
the resulting mixture was stirred at room temperature under
nitrogen overnight. Concentration in vacuo provided an oil which
was diluted with dichloromethane, washed with 1M hydrochloric acid
and 1 M aqueous sodium hydroxide, then dried over anhydrous
magnesium sulfate. Filtration and concentration in vacuo afforded
the compound of Formula 3.0 (+-isomer) (0.095 g, 74%, mp 135-142
.degree. C.).
EXAMPLE 4
[0330] 170
[0331] To the compound of Formula 3.0 (Example 3) (0.09 g)
dissolved in anhydrous dichloromethane (10 mL) was added
trifluoroacetic acid (1 mL) and the resulting solution was stirred
at room temperature for 1 hour. 50% Aqueous sodium hydroxide was
added slowly followed by dichloromethane and brine. The mixture was
shaken well, the organiic phase was separated and dried over
anhydrous magnesium sulfate. Filtration and concentration in vacuo
afforded the compound of Formula 17.0 (+-isomer) (0.053 g, 68%, mp
122.7-128.degree. C.).
EXAMPLE 5
[0332] 171
[0333] The compound of Formula 28.0 (Preparative Example 8) (0.10
g, 0.17 mmol) was combined with 1-hydroxybenzotriazole hydrate
(0.045 g, 0.34 mmol),
1-(3-dimethylaminopropyl)-3-ethylcarbo-diimide hydrochloride (0.064
g, 0.34 mmol), N-tert-butoxy-carbonyl-D-alanine (0.064 g, 0.34
mmol) and anhydrous dimethylformamide (10 mL) and the resulting
mixture was stirred at room temperature under nitrogen overnight.
Concentration in vacuo provided an oil which was diluted with
dichloromethane, washed with IM hydrochloric acid and 1 M aqueous
sodium hydroxide, then dried over anhydrous magnesium sulfate.
Filtration and concentration in vacuo afforded the compound of
Formula 4.0 (+-isomer) (0.104 g, 81%, mp 135.1-142.3.degree.
C.).
EXAMPLE 6
[0334] 172
[0335] To the compound of Formula 4.0 (Example 5) (0.10 g)
dissolved in anhydrous dichloromethane (10 mL) was added
trifluoroacetic acid (1 mL) and the resulting solution was stirred
at room temperature for 1 hour. 50% Aqueous sodium hydroxide was
added slowly followed by dichloromethane and brine. The mixture was
shaken well, the organic phase was separated and dried over
anhydrous magnesium sulfate. Filtration and concentration in vacuo
afforded the compound of formula 18.0 (+-isomer) (0.056 g, 64%, mp
103.degree. C. (DEC)).
EXAMPLE 7
[0336] 173
[0337] The compound of Formula 5.0 (+-isomer) was prepared,
according to procedures similar to those of Examples 1, 3, and 5,
by reacting the compound of Formula 28.0 (Preparative Example 8)
with the amino acid N-tert-butoxycarbonyl-L-phenylalanine. Yield:
76%, mp: 128.6-134.degree. C.
EXAMPLE 8
[0338] 174
[0339] The compound of Formula 6.0 (+-isomer) was prepared,
according to procedures simllar to those of Examples 1, 3, and 5,
by reacting the compound of Fonnula 28.0 (Preparative Example 8)
with the amino acid N-(alpha)-tert-butoxycarbonyl-L-histidine.
Yield: 32%, mp: 96.0-99.7.degree. C.
EXAMPLE 9
[0340] 175
[0341] The compound of Formula 7.0 (+-isomer) was prepared,
according to procedures similar to those of Examples 1, 3, and 5,
by reacting the compound of Formula 28.0 (Preparative Example 8)
with the amino acid N-(alpha)-tert-butoxycarbonyl-L-proline. Yield:
52%, mp: 110.degree. C.
EXAMPLE 10
[0342] 176
[0343] The compound of Formula 8.0 (+-isomer) was prepared,
according to procedures similar to those of Examples 2, 4, and 6,
from the compound of Formula 5.0 (Example 7). Yield: 70%, mp:
116-119.degree. C.
EXAMPLE 11
[0344] 177
[0345] The compound of Formula 9.0 (+-isomer) was prepared,
according to procedures similar to those of Examples 2, 4, and 6,
from the compound of Formula 6.0 (Example 8). Yield: 51%, mp:
101.degree. C.
EXAMPLE 12
[0346] 178
[0347] The compound of Formula 10.0 (+-isomer) was prepared,
according to procedures similar to those of Examples 2, 4, and 6,
from the compound of Formula 7.0 (Example 9). Yield: 46%, mp:
131.6.degree. C.
EXAMPLE 13
[0348] 179
[0349] To the compound of Formula 28.0 (Preparative Example 8)
(0.51 g, 0.85 mmol) and triethylamine (0.18 mL, 1.3 mmol) dissolved
in anhydrous dichloromethane (50 mL) was added ClCH.sub.2C(O)Cl
(chloroacetyl chloride) (0.28 mL, 1.2 eq) dissolved in
dichloromethane (10 mL) at 0.degree. C. After stirring for 1.5
hours, 1 M hydrochloric acid was added and the mixture was shaken.
The organic phase was separated and washed with 1 N aqueous sodium
hydroxide, then brine, and dried over anhydrous magnesium sulfate.
Filtration and concentration in vacuo afforded the compound of
Formula 31.0 (0.58 g, 100%, mp 124.0-134.5.degree. C.). 180
[0350] The compound of Formula 31.0 (0.12 g, 0.18 mmol), morpholine
(5 mL) and anhydrous sodium carbonate (0.038 g, 2 eq) were stirred
at 130.degree. C. overnight. After concentration in vacuo, the
residue was diluted with dichloromethane, washed with water and
dried over anhydrous magnesium sulfate. Filtration and
concentration in vacuo afforded a yellow residue (0.17 g) which was
purified by preparative plate chromatography (silica gel) using 5%
methanol-dichloromethane and concentrated ammonium hydroxides to
provide the compound of Formula 13.0 (0.096 g, 75%, mp
116.6.degree. C.).
EXAMPLE 14
[0351] 181
[0352] The compound of Formula 31.0 (Example 13) (0.12 g, 0.18
mmol), anhydrous dimethylformamide (10 mL), imidazole (0.037 g,
0.54 mmol) and anhydrous sodium carbonate (0.057 g, 0.54 mmol) were
stirred at 130.degree. C. overnight. The mixture was cooled to room
temperature, diluted with water, filtered and the solids washed
with water. The solids were diluted with dichloromethane, washed
with water and then with 1 N aqueous sodium hydroxide. The organic
phase was separated, dried over anhydrous magnesium sulfate,
filtered and concentrated in vacuo to afford a solid (0.084 g)
which was purified by preparative plate chromatography (silica gel)
using 5% methanol-dichloromethane and concentrated ammonium
hydroxide to provide the compound of Formula 14.0 (0.06 g, 48%, mp
148.9.degree. C.).
EXAMPLE 15
[0353] 182
[0354] The compound of Formula 28.0 (Preparative Example 8) (0.21
g, 0.34 mmol) dissloved in anhydrous dichloromethane (10 mL) was
added to a dichloromethane solution (10 mL) of oxallyl chloride
(1.0 mL) and pyridine (0.08 mL, 3 eq) at 0.degree. C. After
stirring the resulting solution for 5 min, concentrated ammonium
hydroxide was added and the mixture was allowed to stir overnight.
The mixture was diluted with dichloromethane and water, shaken and
then the phases were separated. The organic phase was washed with
brine, then with 1 M hydrochloric acid, 1 N aqueous sodium
hydroxide and brine. The organic phase was separated, dried over
anhydrous magnesium sulfate, filtered and concentrated in vacuo to
afford a solid (0.17 g) which was purified by preparative plate
chromatography (silica gel) using 5% methanol-dichloromethane and
concentrated ammonium hydroxide to provide the compound of Formula
15.0 (0.086 g, 37%, mp 152.8.degree. C.).
EXAMPLE 16
[0355] 183
[0356] To the compound of Formulh 16.0 (Example 2) (0.10 g)
dissolved in anhydrous dichloromethane (10 mL) was added
triethylamine (0.032 mL, 1.5 eq)) and methanesulfonyl chloride
(0.014 mL, 1.2 eq) and the resulting solution was stirred at room
temperature overnight. The solution was diluted with
dichloromethane and washed with 1M hydrochloric acid and then with
1 M aqueous sodium hydroxide. The organic phase was separated and
dried over anhydrous magnesium sulfate. Filtration and
concentration in vacuo afforded the compound of Formula 11.0 (0.099
g, 89%, mp 116.degree. C.).
EXAMPLE 17
[0357] 184
[0358] To the compound of Formula 16.0 (Example 2) (0.07 g)
dissolved in anhydrous dichloromethane (10 mL) was added
triethylamine (0.022 mL, 1.5 eq)) and benzoyl chloride (0.014 mL,
1.2 eq) and the resulting solution was stirred at room temperature
overnight. The solution was diluted with dichloromethane and washed
with 1M hydrochloric acid and then with 1 M aqueous sodium
hydroxide. The organic phase was separated and dried over anhydrous
magnesium sulfate. Filtration and concentration in vacuo afforded
the compound of formula 12.0 (0.066 g, 85%, mp 117.2.degree.
C.).
EXAMPLE 18
[0359] 185
[0360] Dissolve 2 g (15 mmol) of methyl 3-(dimethyl amino)
propionate in 20 mL of EtOH and then add 20 mL of 1M LiOH. Stir the
reaction mixture at room temperature for 16 h. Strip off the
solvents. Dissolve the resulting material in water and adjust pH to
.about.6. Concentrate the reaction mixture to give the product.
Mass Spec.: MH.sup.+=118.
EXAMPLE 19
(+)-4-(3.
10-DIBROMO-8-CHLORO-6.11-DIHYDRO-5H-BENZO[5,6]CYCLOHEPTA[1,2-b]P-
YRIDIN-11-YL)-1-[4-(DIMETHYLAMINO)-1-OXOBUTYL]-4-PIPERIDINYL]ACETYL]-PIPER-
IDINE
[0361] 186
[0362] Dissolve 0.1 g (0.23 mmol) of the product of Preparative
Example 8 187
[0363] in 8 mL DMF, add 0.04 g (0.22 mmol) 4-(dimethylamino)
butyric acid hydrochloride, 0.04 g (0.22 nimol) of DEC, 0.03 g
(0.22 mmol) of HOBT and 0.1 mL of N-methyl morpholine at about 0 to
about 4.degree. C. Stir the reaction mixture overnight letting it
warm to room temperature. Remove all the volatiles and then
partition between H.sub.2O-CH.sub.2Cl.sub.2. Extract the aqueous
phase with CH.sub.2Cl.sub.2. Combine the CH.sub.2Cl.sub.2 fractions
and dry over MgSO.sub.4 and concentrate. Purify by flash
chromatography, first eluting with 5%
MeOH-(NH.sub.3)-CH.sub.2Cl.sub.2 and then 10%
MeOH-(NH.sub.3)-CH.sub.2Cl.- sub.2 to obtain the compound of
Formula 12.2. Mass Spec. MH+=709, mp=69-71 .degree. C.
EXAMPLE 20
(+)-4-(3,10-DIBROMO-8-CHLORO-6,11-DIHYDRO-5H-BENZO[5,6]CYCLOHEPTA[1,2-b]PY-
RIDIN-11-YL)-1-[4-(DIMETHYLAMINO)-1-OXOPROPYL]-4-PIPERIDINYL]ACETYL]-PIPER-
IDINE
[0364] 188
[0365] By following essentially the same procedure as described in
Example 19 above, but using 3-(dimethylamino) propionic acid
(Example 18) instead of 4-(dimethylamino) butyric acid
hydrochloride, the compound of Formula 12.3 was prepared.
FAB-MS-MH+=695, mp=82-84.degree. C.
EXAMPIE 21
(+)-4-(3,10-DIBROMO-8-CHLORO-6,11-DIHYDRO-5H-BENZO[5,6]CYCLOHEPTA[1,2-b]PY-
RIDIN-11-YL)-1-[4-(DIMETHYLAMINO)-1-OXOETHYL]-4-PIPERIDINYL]
ACETYL]-PIPERIDINE
[0366] 189
[0367] By following essentially the same procedure as described in
Example 19 above, but using N,N-dimethyl glycine instead of
4-(dimethylamino) butyric acid hydrochloride, the compound of
Fromula 12.1 was prepared. FAB-MS-MH+=681, mp=123-124.degree.
C.
EXAMPLE 22
(+)-4-(3,10-DIBROMO-8-CHLORO-6,11-DIHYDRO-5H-BENZO[5,6]CYCLOHEPTA[1,2-b]PY-
RIDIN-11-YL)-1-[4-(PIPERIDINYL)-1-OXOETHYL]-4-PIPERIDINYL]
ACETYL]-PIPERIDINE
[0368] 190
[0369] By following essentially the same procedure as described in
Example 19 above, but using 1-piperidine propionic acid instead of
4-(dimethylamino) butyric acid hydrochloride, the compound of
Formula 14.2 was prepared. FAB-MS: MH.sup.+735, mp=127-128.degree.
C.
EXAMPLE 23
(+)-4-(3,10-DIBROMO-8-CHLORO-6,11-DIHYDRO-5H-BENZO[5,6]CYCLOHEPTA[1,2-b]PY-
RIDIN-11-YL)-1-[4-(TETRAHYDRO-2H-1,4-THIAZIN-4-YL)-1-OXOETHYL
1-1-DIOXIDE]-4-PIPERIDINYL]ACETYL]PIPERIDINE
[0370] 191
[0371] By following essentially the same procedure as described in
Example 19 above, but using thiomorpholine S-dioxide acetic acid
instead of 4-(dimethylamino) butyric acid hydrochloride, the
compound of Formula 14.1 was prepared. mp=140-141.degree. C.
EXAMPLE 24
(+)-
METHYL-4-[2-[4-[(3,10-DIBROMO-8-CHLORO-6,11-DIHYDRO-5H-BENZO[5,6]CYCL-
OHEPTA[1,2-b]PYRIDIN-11-YL)-1-PIPERIDINYL]-2-OXOETHYL]DELTA-OXO-1-PIPERIDI-
NE-PENTANOATE
[0372] 192
[0373] By following essentially the same procedure as described in
Example 19 above, but using monomethyl glutarate instead of
4-(dimethylamino) butyric acid hydrochloride, the compound of
Formula 15.1 was prepared. FAB-MS-MH+=724, mp=101-102.degree. C.
EXAMPLE 25
(+)-
METHYL-4-[2-[4-[(3,10-DIBROMO-8-CHLORO-6,11-DIHYDRO-5H-BENZO[5,6]CYCL-
OHEPTA[1,2-b]PYRIDIN-11-YL)-1-PIPERIDINYL]-2-OXOETHYL]-GAMMA-OXO-1-PIPERID-
INE-BUTANOATE
[0374] 193
[0375] By following essentially the same procedure as described in
Example 19 above, but using monomethyl succinate instead of
4-(dimethylamino) butyric acid hydrochloride, the compound of
Formula 15.2 was prepared. FAB-MS-MH.sup.+=710, mp=114-115 .degree.
C.
EXAMPLE 26
(+)-
ETHYL-4-[2-[4-[(3,10-DIBROMO-8-CHLORO-6,11-DIHYDRO-5H-BENZO[5,6]CYCLO-
HEPTA[1,2-b]PYRIDIN-11-YL)-1-PIPERIDINYL]-2-OXOETHYL]-BETA-OXO-1-PIPERIDIN-
E-BUTANOATE
[0376] 194
[0377] By following essentially the same procedure as Example 19
above, but using monoethyl malonate instead of 4-(dimethyl-amino)
butyric acid hydrochloride, the compound of Formula 15.3 was
obtained. FAB-MS-MH.sup.+=710, m.p.=77-78.degree. C.
EXAMPLE 27
[0378] 195
[0379] Dissolve the (+) product of Preparative Example 8, Step D
(0.01 g, 0.017 mmol) in 0.5 mL of DMF, stir at room tempeoature o
and add 0.003 g (0.017 mmol) of DEC, 0.002 g (0.017 mmol) of HOBT
and 0.003 g (0.017 mmole) of the product of Preparative Example 12.
Stir the mixture at room temperature for 18 hr, then concentrate in
vacuo to a residue and partition between ethyl acetate and water.
Wash the organic phase with aqueous sodium bicarbonate solution
then brine. Dry the organic phase over magnesium sulfate, filter
and concentrate in vacuo to a residue. Chromatograph the residue on
silica gel, eluting with dichloromethane (saturated with
ammonia)--methanol (95% -5%) to yield the product (0.01 g) ,is a
white solid. M.p.=84.degree.-90.degree. C., Mass Spec.:
MH+=709.
EXAMPLES 28-60
[0380] Follow the procedure of Example 27 but use the acid shown in
Table 1 below instead of the product of Preparative Example 12 to
obtain the compounds of Formula 1.7 196
[0381] wherein W is defined in Table 1. The Formula number of the
compound formed is given in parenthesis below the W
substituent.
1TABLE 1 mp EX. Acid W (.degree. C.) 28 197 198 140-140.8 29 199
200 127.2-127.7 30 201 202 77.2- 78.1 31 203 204 103.5-106.4 32 205
206 129 (d) 33 207 208 75 34 209 210 145.8-147.7 35 211 212
125.8-127.3 36 213 214 95-143 37 215 216 -- 38 217 218 124-125 39
219 220 204.5 40 221 222 137.4-138 41 223 224 115.8-116.4 42 225
226 -- 43 227 228 -- 44 229 230 113-120 45 231 232 95-100 46 233
234 100-108 47 235 236 192-203 48 237 238 172-190 49 239 240
154-163 50 241 242 129-139 51 243 244 114.5-119.3 52 245 246 87-88
53 247 248 102.7-103.3 54 249 250 145 (d) 55 251 252 129-143 56 253
254 124-132 57 255 256 127-136 58 257 258 74-75 59 259 260 -- 60
261 262 114-115 61 263 264 101.1-101.5
EXAMPLE 62
[0382] 265
[0383] Dissolve the (+) product of Preparative Example 8, Step D
(0.744 g, 1.25 mmol) in 20 mL of dichloromethane containing 0.348
mL (2.5 mmol) of triethylamine, stir at room temperature and add
0.1 mL (1.26 mmol) of chloroacetylchloride. Stir for 10 hr then add
20 mL of 1N HCl. Wash the organic layer with aqueous sodium
bicarbonate, dry over magnesium sulfate, and concentrate under
vacuum to give 0.71 g of the product. 266
[0384] Dissolve 0.120 g (0.78 mmol) of the product from Step A,
0.0365 g (0.535 mmol) of 4-metlhylimidazole and 0.057 g (0.535
mmol) of sodium carbonate in 10 mL of DMF and stir at
120-130.degree. C. for 18 hr. Cool to 25.degree. C. and 30 mL of
water and filter the precipitated solid. Dissolve the solid in 50
mL of dichloromethane and wash with 1N NaOH. Dry the organic layer
over magnesium sulfate and concentrate under vacuum. Chromatograph
the residue on a silica gel TLC plate using
methanol-dichloromethane saturated with ammonia (5-95) to give 0.06
g of the product as a white solid mp=148.9.degree. C.
EXAMPLES 63-75
[0385] Follow the procedure of Exajmple 62, but use the amine shown
in Table 2 below instead of 4-methylimidazole, to obtain the
compounds of Formula 1.7 267
[0386] wherein W is defined in Table 2. The Formula number of the
compound formed is given in parenthesis below the W
substituent.
2TABLE 2 mp EX. Amine W (.degree. C.) 63 268 269 137.8 64 270 271
151.3 65 272 273 154.2 66 274 275 150.3 (d) 67 276 277 168.1 68 278
279 123.4 69 280 281 184-190 70 282 283 178.6 71 284 285 156.1 72
286 287 158.7 73 288 289 130.4 74 290 291 122.8 75 292 293
125.3
EXAMPLE 76
[0387] 294
[0388] Dissolve 1 equivalent of the: product of Example 59
(Compound 104.0-B, Table 1) in methanol containing 1.2 equivalents
of 1N KOH in methanol and stir for 48 hr at 25.degree. C. Acidify
to pH 2 with 1N HCl and extract with dichloromethane. Dry the
organic layer over magnesium sulfate and concentrate under vacuum.
Purify the residue by preparative silica gel TLC using
methanol-dichloromethane-acetic acid (5-94-1) to give the product
as a white solid mp=240. 1.degree. C.
EXAMPLE 77
[0389] 295
[0390] Dissolve 1 equivalent of the prduct of Example 30 (Compound
8.0-B, Table 1) in 95% aqueous ethanol containing 1.1 equivalents
of LiOH and stir for 16 hr at 25.degree. C. Concentrate under
vacuum to give the product as a white solid.
EXAMPLE 78
[0391] 296
[0392] Follow the procedure of Example 77 but use the product of
Example 60 (Compound105.0-B, Table 1) instead of the product of
Example 59 (Compound104.0-B, Table 1) to obtain the product as a
white solid.
EXAMPLE 79
[0393] 297
[0394] Follow the procedure of Example 77 but use the product of
Example 61 (Compound 106.0-B, Table 1) instead of the product of
Example 59 (Compound104.0-B, Table 1) to obtain the product as a
white solid.
EXAMPLE 80
[0395] 298
[0396] Dissolve 1 equivalent of the product of Example 43 (Compound
68.0-B, Table 1) in 95% aqueous methanol containing 1.1 equivalents
of NaOH and stir for 16 hr at 25.degree. C. Concentrate under
vacuum to give the product as a white solid. mp=215.5-216.2.degree.
C
EXAMPLE 81
[0397] 299
[0398] Dissolve 1 equivalent of the prduct of Example 58 (Compound
101.0-B, Table 1) in 95% aqueous methanol containing 1.1
equivalents of NaOH and stir for 16 hr at 25.degree. C. Concentrate
under vacuum to give the product as a white solid. mp=240.degree.
C. (d).
EXAMPLE 82
[0399] 300
[0400] Dissolve 1.0 equivalent of the product of Example 81
(Compound 107.0-B) in DMF containing 5.0 equivalents of ammonium
chloride and 1.0 equivalent each of DEC, HOBT and
N-methylmorpholine. Stir the mixture at room temperature for 18 hr,
then concentrate in vacuo to a residue and partition between ethyl
acetate and water. Wash the organic phase with aqueous sodium
bicarbonate solution then brine. Dry the organic phase over
magnesium sulfate, filter and concentrate in vacuo to a residue.
Chromatograph the residue on silica gel to obtain the product as a
white solid mp=125.5-126.5.degree. C.
EXAMPLE 83
[0401] 301
[0402] Dissolve 1.0 equivalent of the product of Example 78
(Compoundl8.0-B) in DMF containing 5.0 equivalents of ammonium
chloride and 1.0 equivalent each of DEC, HOBT and
N-methylmorpholine. Stir the mixture at room temperature for 18 hr,
then concentrate in vacuo to a residue and partition between ethyl
acetate and water. Wash the organic phase with aqueous sodium
bicarbonate solution then brine. Dry the organic phase over
magnesium sulfate, filter and concentrate in vacuo to a residue.
Chromatograph the residue on silica gel to obtain the product as a
white solid mp=142.8-143.3.degree. C.
EXAMPLE 84
[0403] 302
[0404] Dissolve 1.0 equivalent of the product of Example 80
(Compound 70.0-B) in DMF containing 5.0 equivalents of ammonium
chloride and 1.0 equivalent each of DEC, HOBT and
N-methylmorpholine. Stir the mixture at room temperature for 18 hr,
then concentrate in vacuo to a residue and partition between ethyl
acetate and water. WVash the organic phase with aqueous sodium
bicarbonate solution then brine. Dry the organic phase over
magnesium sulfate, filter and concentrate in vacuo to a residue.
Chromatograph the residue on silica gel to obtain the product as a
white solid mp=119.2-120.degree. C.
EXAMPLE 85
[0405] 303
[0406] Dissolve 1.0 equivalent of the product of Example 81
(Compound 107.0-B) in DMF containing 5.0 equivalents of ammonium
chloride and 1.0 equivalent each of DEC, HOBT and
N-methylmorpholine. Stir the mixture at room temperature for 18 hr,
then concentrate in vacuo to a residue and partition between ethyl
acetate and water. Wlash the organic phase with aqueous sodium
bicarbonate solution then brine. Dry the organic phase over
magnesium sulfate, filter and concentrate in vacuo to a residue.
Chromatograph the residue on silica gel to obtain the product as a
white solid.
EXAMPLE 86
[0407] 304
[0408] Dissolve 1.0 equivalent of the product of Example 79
(Compound19.0-B) in DMF containing 5.0 equivalents of ammonium
chloride and 1.0 equivalent each of DEC, HOBT and
N-methylmorpholine. Stir the mixture at room temperature for 18 hr,
then concentrate in vacuo to a residue and partition between ethyl
acetate and water. Wash the organic phase with aqueous sodium
bicarbonate solution then brine. Dry the organic phase over
magnesium sulfate, filter and concentrate in vacuo to a residue.
Chromatograph the residue on silica gel to obtain the product as a
white solid.
EXAMPLE 87
[0409] 305
[0410] Dissolve 1.0 equivalent of the product of Example 59
(Compound 104.0-B, Table 1) in dichloromethane containing 4.0
equivalents of anhydrous hydrazine and stir for 48 hr. Concentrate
under vacuum and chromatograph the residue on preparative silica
gel TLC using methanol-dichloromethane 5-95) to yield the product
as a yellow solid, mp=90.degree. C.
EXAMPLE 88
[0411] 306
[0412] Dissolve 1.0 equivalent of the product of Example 59
(Compound 104.0-B, Table 1) in methanol containing 1.4 equivalents
of LiOH and stir for 18 hr. Add DMF containing 1.0 equivalent each
of DEC, HOBT and N-methylmorpholine and
O-tert-butyldimethylsilylhydroxylamine. Stir the mixture at room
temperature for 48 hr, then concentrate in vacuo to a residue and
partition between ethyl acetate and water. Dry the organic phase
over magnesium sulfate, filter and chromatograph on silica gel
using methanol-dichloromethane (5-95) to obtain the product as a
white solid, mp=103.0.degree. C.
EXAMPLE 89
[0413] 307
[0414] Dissolve 1.0 equivalent the product of Example 59 (Compound
104.0-B, Table 1) in methanol containing 3.0 equivalents of KOH and
3.0 equivalent of glycine hydrochloride tert-butyl ester and stir
for 7 days. Concentrate under vacuum and chromatograph the residue
on silica gel using methanol-dichloromethane (5-95 to obtain the
product as a yellow solid, mp=108.degree. C.
EXAMPLE 90
[0415] 308
[0416] Dissolve 1.0 equivalent the product of Example 59 (Compound
104.0-B, Table 1) in methanol containing 3.0 equivalents of KOH and
3.0 equivalent of O-benzylhydroxyl-amine hydrochloride and stir for
48 hr. Concentrate under vacuum and chromatograph the residue on
silica gel using methanol-dichloromethane-acetic acid (10-89.5-0.5)
to obtain the product as a yellow solid mp=75.degree. C.
EXAMPLE 91
[0417] 309
[0418] Dissolve 1.0 equivalent the product of Example 59 (Compound
104.0-B, Table 1) in methanol 4.0 equivalent of methylamine and
stir for 18 hr. Concentrate under vacuum and chromatograph the
residue on silica gel using methanol-dichloromethane saturated with
ammonia (5-95) to obtain the product as a yellow solid
mp=86-132.degree. C.
EXAMPLE 92
[0419] 310
[0420] Dissolve 1.0 equivalent of the product from Example 52
(Compound 86.0-B, Table 1) in dichloromethane containing 2
equivalents of trifluoroacetic acid and stir for 2 hr. Concentrate
under vacuum and partition the residue between dichloro-methane and
aqueous sodium bicarbonate. Dry the organic layer over magnesium
sulfate and concentrate under vacuum to yield the product as a
white solid, mp=120.6-120.8.degree. C.
EXAMPLE 93
[0421] 311
[0422] Dissolve 1.0 equivalent of the product from Example 53
(Compound 89.0-B, Table 1) in dichloromethane containing 2
equivalents of trifluoroacetic acid and stir for 2 hr. Concentrate
under vacuum and partition the residue between dichloro-methane and
aqueous sodium bicarbonate. Dry the organic layer over magnesium
sulfate and concentrate under vacuum to yield the product as a
white solid, mp=114-115.degree. C.
EXAMPLE 94
[0423] 312
[0424] Dissolve 1.0 equivalent of the product from Example 1 in
dichloromethane containing 2 equivalents of trifluoroacetic acid
and stir for 2 hr. Concentrate under vacuum and partition the
residue between dichloro-methane and aqueous sodium bicarbonate.
Dry the organic layer over magnesium sulfate and concentrate under
vacuum. Dissolve the residue in dichloromethane containing 1.5
equivalents of triethyl amine and 1.2 equivalents of
dimethylsulfamoyl chloride. Stir for 18 hr then wash with 1N HCl
followed by 1N NaOH. Dry the organic layer over magnesium sulfate
and concentrate under vacuum to obtain the produce,
mp=124.4-130.degree. C.
EXAMPLE 95
[0425] 313
[0426] Dissolve 1.0 equivalent of the product from Example 1 in
dichloromethane containing 2 equivalents of trifluoroacetic acid
and stir for 2 hr. Concentrate under vacuum and partition the
residue between dichloro-methane and aqueous sodium bicarbonate.
Dry the organic layer over magnesium sulfate and concentrate under
vacuum. Dissolve the residue in 10.0 equivalents of aqueous
sulfamide and reflux for 48 hr. Concentrate under vacuum and
chromatograph the residue on silica gel using
methanol-dichloromethane saturated with ammonia (5-95) to obtain
the product, mp=151.9.degree. C.
EXAMPLE 96
[0427] 314
[0428] The product of Example 31 (Compound16.0-B, Table 1) (372.1
mg, 0.468 mmol) was dissolved in 3 mL of 6 M HCl and the solution
stirred at room temperature overnight. The reaction mixture was
added to 25 mL water, and the resulting precipitate was filtered
and washed with 0.1 M HCl. The filtrate was saturated with NaCl and
extracted continuously for 48 h to provide additional crude
product. Purify the combined crude material by flash chromatoghrapy
(C-18 reverse phase silica, gradient of 50% MeOH/0.17 M HOAC to 90%
MeOH/0.17 M HOAc). The resulting material was dissolved in MeOH and
added to water and the resulting suspension evaporated to dryness
to give the title compound as a white solid (mp
133.50-141.2.degree. C., heating 2.degree.-3.degree. C./min).
EXAMPLE 97
[0429] 315
[0430] The product of Example 34 (Compound 24.0-B, Table 1) (450.0
mg, 0.56 mmol) was dissolved in 20 mL CH.sub.2Cl.sub.2, cooled to
0.degree. C., and 8 mL of trifluoroacetic acid was added slowly.
After 1 h the cold mixture was diluted with 50% NaOH (aq) and
water. The mixture was extracted with CH.sub.2Cl.sub.2, which was
then dried (MgSO.sub.4) and evaporated to give the title compound
as a yellow solid (230 mg, mp 161.0.degree.-163.0.degree. C.).
EXAMPLE 98
[0431] 316
[0432] The product of Example 96 (Compound 74.0-B) (93.6 mg, 0.129
mmol) and 1-hydroxybenzotriazole (27.3 mg, 0.202 mmol) were
dissolved in 1 mL of DMF. NH.sub.4Cl (14.8 mg, 0.276 mmol),
N-methylmorpholine (70 .mu.L) and DEC.multidot.HCl (30.8 mg, 0.161
mmol) were added. After 4 h the mixture was evaporated and the
residue purified by flash chromatoghrapy (C-18 reverse phase
silica, gradient of 50% MeOH/0.17 M HOAc to 90% MeOH/0.17 M HOAc).
The resulting material was lyophilized from HOAc/H.sub.2O to give
the title compound as a tan solid (67.7 mg, mp
115.2.degree.-122.0.degree. C., heating 2.degree.-3.degree.
C./min).
EXAMPLE 99
[0433] 317
[0434] Dissolve 1.0 equivalent of the product of Example 77
(Compound 14.0-B) in DMF containing 1.0 equivalent each, of DEC,
HOBT, N-methylmorpholine and pyrrolidine. Stir the mixture at room
temperature for 18 hr, then concentrate in vacuo to a residue and
partition between ethyl acetate and water. Wash the organic phase
with aqueous sodium bicarbonate solution then brine. Dry the
organic phase over magnesium sulfate, filter and concentrate in
vacuo to a residue. Chromatograph the residue on silica gel to
obtain the product as a white solid.
EXAMPLE 100
[0435] 318
[0436] Dissolve 1.0 equivalent of the product of Example 77
(Compound 14.0-B) in DMF containing 1.0 equivalent each of DEC,
HOBT, N-methylmorpholine and piperidine. Stir the mixture at room
temperature for 18 hr, then concentrate in vacuo to a residue and
partition between ethyl acetate and water. Wash the organic phase
with aqueous sodium bicarbonate solution then brine. Dry the
organic phase over magnesium sulfate, filter and concentrate in
vacuo to a residue. Chromatograph the residue on silica gel to
obtain the product as a white solid mp=135-136.degree. C.
EXAMPLE 101
[0437] 319
[0438] Dissolve 1.0 equivalent of the product of Example 77
(Compound 14.0-B) in DMF containing 1.0 equivalent each of DEC,
HOBT, N-methylmorpholine and morpholine. Stir the mixture at room
temperature for 18 hr, then concentrate in vacuo to a residue and
partition between ethyl acetate and water. Wash the organic phase
with aqueous sodium bicarbonate solution then brine. Dry the
organic phase over magnesium sulfate, filter and concentrate in
vacuo to a residue. Chromatograph the residue on silica gel to
obtain the product as a white solid mp=135-136.degree. C.
EXAMPLE 102
[0439] 320
[0440] Dissolve 1.0 equivalent of the product of Example 77
(Compound 14.0-B) in DMF containing 1.0 equivalent each of DEC,
HOBT, N-methylmorpholine and dimethylamine. Stir the mixture at
room temperature for 18 hr, then concentrate in vacuo to a residue
and partition between ethyl acetate and water. Wash the organic
phase with aqueous sodium bicarbonate solution then brine. Dry the
organic phase over magnesium sulfate, filter and concentrate in
vacuo to a residue. Chromatograph the residue on silica gel to
obtain the product as a white solid mp=133-134.degree. C.
[0441] Assays
[0442] FPT IC.sub.50 (inhibition of farnesyl protein transferase,
in vitro enzyme assay) and COS Cell IC.sub.50 (Cell-Based Assay)
were determined following the assay procedures described in WO
95/10516, published Apr. 20, 1995. GGPT IC.sub.50 (inhibition of
geranylgeranyl protein transferase, in vitro enzyme assay), Cell
Mat Assay, and anti-tumor activity (in vivo anti-tumor studies)
could be determined by the assay procedures described in WO
95/10516. The disclosure of WO 95/10516 is incorporated herein by
reference thereto.
[0443] Additional assays can be carried out by following
essentially the same procedure as described above, but with
substitution of alternative indicator tumor cell lines in place of
the T24-BAG cells. The assays can be conducted using either
DLD-1-BAG human colon carcinoma cells expressing an activated K-ras
gene or SW620-BAG human colon carcinoma cells expressing an
activated K-ras gene. Using other tumor cell lines known in the
art, the activity of the compounds of this invention against other
types of cancer cells could be demonstrated.
[0444] Soft Agar Assay
[0445] Anchorage-independent growth is a characteristic of
tumorigenic cell lines. Human tumor cells are suspended in growth
medium containing 0.3% agarose and an indicated concentration of a
farnesyl transferase inhibitor. The solution is overlayed onto
growth medium solidified with 0.6% agarose containing the same
concentration of farnesyl transferase inhibitor as the top layer.
After the top layer is solidified, plates are incubated for 10-16
days at 37.degree. C. under 5% CO.sub.2 to allow colony outgrowth.
After incubation, the colonies are stained by overlaying the agar
with a solution of MTr (3-[4,5-dimethyl-thiazol-2-yl]-
-2,5-diphenyltetrazolium bromide, Thiazolyl blue) (1 mg/mL in PBS).
Colonies can be counted and the IC.sub.50's can be determined.
[0446] The results are given in Table 3. In Table 3 "Ex. No."
stands for "Example Number".
3TABLE 3 FPT IC.sub.50 (nM) Ex No. Formula No. (H-ras) COS Cell
IC.sub.50 (nM) 1 2.0 12.3 -- 2 11.0 5 30 3 3.0 6.3 -- 4 12.0 5.2
140 5 4.0 8.4 -- 6 13.0 8.1 39 7 5.0 31.7 -- 8 6.0 7.4 125 9 7.0
11.6 700 10 8.0 8.3 300 11 9.0 15 -- 12 10.0 4.4 55 19 12.2 35 --
20 12.3 5.8 10 21 12.1 2.5 10 22 14.2 <2.6 43 23 14.1 2.7 14 24
15.1 4.8 10 25 15.2 5.1 39 26 15.3 <2 125
[0447] Additional results are given in Table 4. "Cmpd" represents
"compound" and the number in that column is the compound or
structure number. The FPT IC.sub.50 results are expressed as
nanomolar (nM) in Table 4.
4TABLE 4 Cmpd FPT IC.sub.50 Cmpd FPT IC.sub.50 Cmpd FPT IC.sub.50
6.0-B 2.7 7.0-B <2.6 8.0-B 2.8 9.0-B 5.8 10.0-B 2.5 11.0-B 1.6
12.0-B 3.0 13.0-B 4.1 14.0-B 0.9, 1.9 16.0-B 5.3 17.0-B 1.2 18.0-B
1.2 19.0-B 1.1 20.0-B 5.6 21.0-B 5.7 22.0-B 1.8 23.0-B 2.8 24.0-B
0.7 26.0-B 3.2 30.0-B 2.9 32.0-B 3.5 33.0-B 2.1 34.0-B 4.2 35.0-B
6.4 37.0-B 2.6 38.0-B 3.2 39.0-B 4.1 44.0-B 2.6 49.0-B 2.1 50.0-B
3.3 51.0-B 8.1 52.0-B 1.3 53.0-B 3.6 54.0-B 3.9 55.0-B 9.2 56.0-B
3.3 57.0-B 1.8 58.0-B 16 59.0-B 4.0 60.0-B 5.4 63.0-B 3.5 64.0-B
5.2 67.0-B 3.5 68.0-B 6.9 69.0-B 2.5 70.0-B 1.4 71.0-B 3.3 72.0-B
5.4 74.0-B 1.7 75.0-B 4.7 76.0-B 6.7 77.0-B 3.2 79.0-B 4.4 81.0-B
9.3 82.0-B 7.1 85.0-B 5.0 86.0-B 18 88.0-B 2.2 89.0-B 8.7 90.0-B
3.8 92.0-B 2.2 95.0-B 8.3 101.0-B 5.3 107.0-B 3.6 114.0-B 3.9
114.2-B 3.9 114.3-B 7.2 114.4-B 7.8 -- --
[0448] The following compounds had the following FPT IC.sub.50
(H-ras) results: 79.0-B, 4.4 nM; 23.0-B, 2.8 nM; 104.0-B, 4.6 nM,
108.0-B, 3.5 nM; 16.0, 5 nM; 17.0, 5.2 nM; and 18.0, 8.1 nM.
[0449] The following compounds had the following FPTR IC.sub.50
(K-ras) results: 2.0, 43.2 nM: 3.0, 3OnM; 4.0, 22 nM; 6.0, 4lnM;
8.0, 71.2 nM: 9.0, 49 nM; 12.1, 19.4 nM; 12.3, 19.8 nM; 7.0, 34.8
nM; 10.0, 35 nM; 6.0-B, 14.5 nM; 9.0-B, 41 nM; 10.0-B, 15.8 nM;
11.0-B, 12.6 nM; 12.0-B, 26.7 nM; 13.0-B, 19.7 nM; 7.0-B, 15.4 nM;
16.0, 20.5 nM; 17.0, 23.8 nM; and 18.0, 31 nM.
[0450] Additional results are given in Table 5. "Cmpd" represents
"compound" and the number in that column is the compound or
structure number. The FPTY IC.sub.50, COS Cell IC.sub.50 and Soft
Agar IC.sub.50 results are expressed as nanomolar (nM) in Table
5.
5TABLE 5 Cmpd Cos Cell Soft Agar Cmpd Cos Cell Soft Agar 6.0 125 --
12.1 -- 160, 90 12.3 -- 190 88.0-B 23 >500 89.0-B 550 >500
10.0 -- 250 24.0-B 100 >500 63.0-B 55 500 6.0-B 30 210 9.0-B 25
>500 10.0-B 20 >300 11.0-B 29 >500 12.0-B 20 >500
13.0-B 25 >500 16.0-B 16 90 21.0-B -- >500 22.0-B 35 19.5,
26.0-B 30 >500 27.5 30.0-B 9 500 34.0-B 9 460 35.0-B 30, 50 300
37.0-B 100, <10 75 38.0-B 10 210 39.0-B 16 220 44.0-B <10 120
52.0-B 400 500 53.0-B 50 >500 54.0-B 100 >500 55.0-B 200
>500 56.0-B 50 >500 57.0-B <10 110 75.0-B 10 150 77.0-B 14
240, 214 79.0-B 150 >500 82.0-B <10 >500 85.0-B 14 333
95.0-B 11 >500 114.2-B 10 75 114.3-B 11 432 114.4-B 10 363 7.0-B
43 -- 23.0-B <10 >500 17.0-B >1000 >500 32.0-B 28
>500 33.0-B 10 >120 49.0-B 38 >500 59.0-B 50 500 60.0-B
120 500 64.0-B 10 130 72.0-B 10 320 81.0-B 28 >500 104.0-B 40
>500 15.2 -- 250 14.0-B 250, 200 >500 18.0-B 75 >500
19.0-B 17.5, <10 440, 500, 275, 197 50.0-B <10 95 51.0-B 280
-- 68.0-B 10 220 69.0-B 25 190 70.0-B 25 250 71.0-B 19 240 74.0-B
40 500 92.0-B 15 45 101.0-B 10 320 107.0-B 30 480 108.0-B 30 --
114.0-B 35 395 16.0 30 75 17.0 140 -- 18.0 39 200
[0451] For preparing pharmaceutical compositions from the compounds
described by this invention, inert, pharmaceutically acceptable
carriers can be either solid or liquid. Solid form preparations
include powders, tablets, dispersible granules, capsules, cachets
and suppositories. The powders and tablets may be comprised of from
about 5 to about 70 percent active ingredient. Suitable solid
carriers are known in the art, e.g. magnesium carbonate, magnesium
stearate, talc, sugar, lactose. Tablets, powders, cachets and
capsules can be used as solid dosage forms suitable for oral
administration.
[0452] For preparing suppositories, a low melting wax such as a
mixture of fatty acid glycerides or cocoa butter is first melted,
and the active ingredient is dispersed homogeneously therein as by
stirring. The molten homogeneous mixture is then poured into
convenient sized molds, allowed to cool and thereby solidify.
[0453] Liquid form preparations include solutions, suspensions and
emulsions. As an example may be mentioned water or water-propylene
glycol solutions for parenteral injection.
[0454] Liquid form preparations may also include solutions for
intranasal administration.
[0455] Aerosol preparations suitable for inhalation may include
solutions and solids in powder form, which may be in combination
with a pharmaceutically acceptable carrier, such as an inert
compressed gas.
[0456] Also included are solid form preparations which are intended
to be converted, shortly before use, to liquid form preparations
for either oral or parenteral administration. Such liquid forms
include solutions, suspensions and emulsions.
[0457] The compounds of the invention may also be deliverable
transdermally. The transdermal compositions can take the form of
creams, lotions, aerosols and/or emulsions and can be included in a
transdermal patch of the matrix or reservoir type as are
conventional in the art for this purpose.
[0458] Preferably the compound is administered orally.
[0459] Preferably, the pharmaceutical preparation is in unit dosage
form. In such form, the preparation is subdivided into unit doses
containing appropriate quantities of the active component, e.g., an
effective amount to achieve the desired purpose.
[0460] The quantity of active compound in a unit dose of
preparation may be varied or adjusted from about 0.1 mg to 1000 mg,
more preferably from about 1 mg. to 300 mg, according to the
particular application.
[0461] The actual dosage employed may be varied depending upon the
requirements of the patient and the severity of the condition being
treated. 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.
[0462] The amount and frequency of administration of the compounds
of the invention and the pharmaceutically acceptable salts thereof
will be regulated according to the judgment of the attending
clinician considering such factors as age, condition and size of
the patient as well as severity of the symptoms being treated. A
typical recommended dosage regimen is oral administration of from
10 mg to 2000 mg/day preferably 10 to 1000 mg/day, in two to four
divided doses to block tumor growth. The compounds are non-toxic
when administered within this dosage range.
[0463] The following are examples of pharmaceutical dosage forms
which contain a compound of the invention. The scope of the
invention in its pharmaceutical composition aspect is not to be
limited by the examples provided.
[0464] Pharmaceutical Dosage Form Examples
EXAMPLE A
Tablets
[0465]
6 Tablets No. Ingredients mg/tablet mg/tablet 1. Active compound
100 500 2. Lactose USP 122 113 3. Corn Starch, Food Grade, 30 40 as
a 10% paste in Purified Water 4. Corn Starch, Food Grade 45 40 5.
Magnesium Stearate 3 7 Total 300 700
[0466] Method of Manufacture
[0467] Mix Item Nos. 1 and 2 in a suitable mixer for 10-15 minutes.
Granulate the mixture with Item No. 3. Mill the damp granules
through a coarse screen (e.g., 1/4", 0.63 cm) if necessary. Dry the
damp granules. Screen the dried granules if necessary and mix with
Item No. 4 and mix for 10-15 minutes. Add Item No. 5 and mix for
1-3 minutes. Compress the mixture to appropriate size and weigh on
a suitable tablet machine.
EXAMPLE B
Capsules
[0468]
7 Capsules No. Ingredient mg/capsule mg/capsule 1. Active compound
100 500 2. Lactose USP 106 123 3. Corn Starch, Food Grade 40 70 4.
Magnesium Stearate NF 7 7 Total 253 700
[0469] Method of Manufacture
[0470] Mix Item Nos. 1, 2 and 3 in a suitable blender for 10-15
minutes. Add Item No. 4 and mix for 1-3 minutes. Fill the mixture
into suitable two-piece hard gelatin capsules on a suitable
encapsulating machine.
[0471] While the present invention has been described in
conjunction with the specific embodiments set forth above, many
alternatives, modifications and variations thereof will be apparent
to those of ordinary skill in the art. All such alternatives,
modifications and variations are intended to fall within the spirit
and scope of the present invention.
* * * * *