Methionine aminopeptidase-2 inhibitors and methods of use thereof Number:7,105,482 from the United States Patent and Trademark Office (PTO) owispatent The present invention provides methods of treating, parasitic infections, thymoma, and lymphoid malignancies in a subject by administering to the subject a therapeutically effective amount of one or more of the compounds of the invention.<H aminopeptidase, inhibitors, Methionine, amin, 7105482.html, Patent, pto, 7105482

Title: Methionine aminopeptidase-2 inhibitors and methods of use thereof

Abstract: The present invention provides methods of treating, parasitic infections, thymoma, and lymphoid malignancies in a subject by administering to the subject a therapeutically effective amount of one or more of the compounds of the invention.

Patent Number: 7,105,482 Issued on 09/12/2006 to Olson, et al.

Inventors: Olson; Gary L. (Mountainside, NJ), Self; Christopher (West Caldwell, NJ), Lee; Lily (Edison, NJ), Cook; Charles Michael (Mendham, NJ), Birktoft; Jens (New York, NY), Morgan; Barry (Franklin, MA), Arico-Muendel; Christopher C. (West Roxbury, MA)
Assignee: Praecis Pharmaceuticals, Inc. (Waltham, MA)

Appl. No.: 10/429,174

Filed: May 2, 2003

Related U.S. Patent Documents


Application Number	Filing Date	Patent Number	Issue Date
10138935	May., 2002
10001945	Nov., 2001
09972772	Oct., 2001
09704251	Nov., 2000	6548477

Current U.S. Class: 514/2 ; 514/16; 514/475; 514/478; 514/588

Current International Class: A61K 31/336 (20060101); A61K 38/08 (20060101)

Field of Search: 530/300,329,350 544/386,391 546/224,244 548/567 549/332 560/157,159 564/32 514/2,16,255.01,315,317,326,428,475,478,533

References Cited [Referenced By]

U.S. Patent Documents


4703107	October 1987	Monsigny et al.
5135919	August 1992	Folkman et al.
5180735	January 1993	Kishimoto et al.
5180738	January 1993	Kishimoto et al.
5290807	March 1994	Folkman et al.
5422363	June 1995	Yanai et al.
5648382	July 1997	Billington et al.
5698586	December 1997	Kishimoto et al.
5767293	June 1998	Oku et al.
5789405	August 1998	Oku et al.
6017954	January 2000	Folkman et al.
6086865	July 2000	Folkman et al.
6548477	April 2003	Olson et al.
6919307	July 2005	Olson et al.
2002/0193298	December 2002	Olson et al.

Foreign Patent Documents


0 354 767	Feb., 1990	EP
0 357 061	Mar., 1990	EP
0 359 036	Mar., 1990	EP
0 387 650	Sep., 1990	EP
0 415 294	Mar., 1991	EP
0 415 294	Mar., 1991	EP
WO 98/13059	Apr., 1998	WO
WO 98/56372	Dec., 1998	WO
WO 9959986	Nov., 1999	WO
WO 9961432	Dec., 1999	WO
WO 00/64486	Nov., 2000	WO
WO 00/64486	Nov., 2000	WO
WO 0069472	Nov., 2000	WO

Other References

Denmeade SR, et al. "Enzymatic activation of a doxorubicin-peptide prodrug by prostate-specific antigen." Cancer Res. Jun. 15, 1998;58(12):2537-40. cited by other .
Griffith EC, et al. "Molecular recognition of angiogenesis inhibitors fumagillin and ovalicin by methionine aminopeptidase 2." Proc Natl Acad Sci U S A. Dec. 22, 1998;95(26):15183-8. cited by other .
Griffith EC, et al. "Methionine aminopeptidase (type 2) is the common target for angiogenesis inhibitors AGM-1470 and ovalicin". Chem Biol. Jun. 1997;4(6):461-71. cited by other .
Han CK, et al. "Design and synthesis of highly potent fumagillin analogues from homology modeling for a human MetAP-2." Bioorg Med Chem Lett. Jan 3, 2000;10(1):39-43. cited by other .
Landquist, JK "Some Degradation Products of Fumagillin" J. Chem. Soc. 1956: 4237-4245. cited by other .
Tarbell et al. "Chemical Evidence For At Least Two Different Precursors To Cyclohexene Formed By Electron Irradiation Of Cyclohexane" Feb. 20, 1960; 82:1005-1007. cited by other .
Liu S, et al. Structure of human methionine aminopeptidase-2 complexed with fumagillin. Science. Nov. 13, 1998;282(5392):1324-7. cited by other .
Sin N, et al. "The anti-angiogenic agent fumagillin covalently binds and inhibits the methionine aminopeptidase, MetAP-2." Proc Natl Acad Sci U S A. Jun. 10, 1997;94(12):6099-103. cited by other .
Timar F, et al. "The antiproliferative action of a melphalan hexapeptide with collagenase-cleavable site." Cancer Chemother Pharmacol. 1998;41(4):292-8. cited by other .
Turk BE, et al. "Selective inhibition of amino-terminal methionine processing by TNP-470 and ovalicin in endothelial cells." Chem Biol. Nov. 1999;6(11):823-33. cited by other .
De Marre, A. et al. "Synthesis and evaluation of macromolecular prodrugs of mitomycin C," Journal of Controlled Release 36(1/2):87-97 (Sep. 1995). cited by other .
Nichifor, M. et al. "Chemical and enzymatic hyrdolysis of dipeptide derivatives of 5-fluorouracil," Journal of Controlled Release 47:271-281 (1997). cited by other.

Primary Examiner: Russel; Jeffrey Edwin
Attorney, Agent or Firm: Lahive & Cockfield, LLP DeConti, Jr.; Giulio A. Zacharakis; Maria Laccotripe

Parent Case Text

RELATED APPLICATION

This application is a continuation-in-part of U.S. patent application Ser. No. 10/138,935, filed May 2, 2002 which is a continuation-in-part of U.S. patent application Ser. No. 10/001,945, filed Nov. 1, 2001, pending; which in turn is a continuation-in-part of U.S. patent application Ser. No. 09/972,772, filed Oct. 5, 2001, pending; which in turn is a continuation-in-part of U.S. patent application Ser. No. 09/704,251, filed Nov. 1, 2000, now U.S. Pat. No. 6,548,477. The entire contents of each of the aforementioned applications are hereby incorporated by reference.

Claims

We claim:

1. A method of treating a subject having a lymphoid malignancy, comprising administering to the subject a therapeutically effective amount of a compound comprising the structure of Formula I, ##STR00057## wherein A is a Met-AP2 inhibitory core; W is O or NR.sub.2; R.sub.1 and R.sub.2 are each, independently, hydrogen or alkyl; X is alkylene or substituted alkylene; n is 0 or 1; R.sub.3 and R.sub.4 are each, independently, hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl; or R.sub.3 and R.sub.4, together with the carbon atom to which they are attached, form a carbocyclic or heterocyclic group; or R.sub.3 and R.sub.4 together form an alkylene group; Z is --C(O)-- or alkylene-C(O)--; and P is a peptide comprising from 1 to about 100 amino acid residues attached at its amino terminus to Z or a group OR.sub.5 or N(R.sub.6)R.sub.7, wherein R.sub.5, R.sub.6 and R.sub.7 are each, independently, hydrogen, alkyl, substituted alkyl, azacycloalkyl or substituted azacycloalkyl; or R.sub.6 and R.sub.7, together with the nitrogen atom to which they are attached, form a substituted or unsubstituted heterocyclic ring structure; or Z is --O--, --NR.sub.8--, alkylene-O-- or alkylene-NR.sub.8--, where R.sub.8 is hydrogen or alkyl; and P is hydrogen, alkyl or a peptide consisting of from 1 to about 100 amino acid residues attached at its carboxy terminus to Z.

2. The compound of claim 1, wherein at least one of R.sub.1, R.sub.3 and R.sub.4 is a substituted or unsubstituted alkyl group.

3. The compound of claim 2, wherein at least one of R.sub.1, R.sub.3 and R.sub.4 is a substituted or unsubstituted normal, branched or cyclic C.sub.1 C.sub.6 alkyl group.

4. The compound of claim 3, wherein at least one of R.sub.1, R.sub.3 and R.sub.4 is a normal or branched C.sub.1 C.sub.4 alkyl group.

5. The compound of claim 1, wherein one of R.sub.3 and R.sub.4 is a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted heteroarylalkyl group, or a substituted or unsubstituted aryl alkyl group.

6. The compound of claim 5, wherein one of R.sub.3 and R.sub.4 is selected from the group consisting of phenyl, naphthyl, indolyl, imidazolyl, pyridyl, benzyl, naphthylmethyl, indolylmethyl, imidazolylmethyl and pyridylmethyl.

7. The compound of claim 1, wherein n is 1 and X is C.sub.1 C.sub.6-alkylene.

8. The compound of claim 7, wherein X is methylene or ethylene.

9. The compound of claim 1, wherein Z is C.sub.1 C.sub.6-alkylene-C(O)--.

10. The compound of claim 9, wherein Z is methylene-C(O)-- or ethylene-C(O)--.

11. The compound of claim 1, wherein at least one of R.sub.6 and R.sub.7 is alkyl, substituted alkyl, substituted or unsubstituted azacycloalkyl or substituted or unsubstituted azacycloalkylalkyl.

12. The compound of claim 11, wherein at least one of R.sub.6 and R.sub.7 is an azacycloalkyl group having an N-alkyl substituent.

13. The compound of claim 12, wherein the N-alkyl substituent is a C.sub.1 C.sub.4-alkyl group.

14. The compound of claim 13, wherein the N-alkyl substituent is a methyl group.

15. The compound of claim 1, wherein R.sub.6 and R.sub.7, together with the nitrogen atom to which they are attached, form a substituted or unsubstituted five or six-membered aza- or diazacycloalkyl group.

16. The compound of claim 15, wherein R.sub.6 and R.sub.7, together with the nitrogen atom to which they are attached, form a substituted or unsubstituted five or six-membered diazacycloalkyl group which includes an N-alkyl substituent.

17. The compound of claim 16, wherein the N-alkyl substituent is a C.sub.1 C.sub.4-alkyl group.

18. The compound of claim 17, wherein the N-alkyl substituent is a methyl group.

19. The compound of claim 1, wherein P is NH2 or one of the groups shown below: ##STR00058##

20. A method of treating a subject having a parasitic infection, comprising administering to the subject a therapeutically effective amount of a compound comprising the structure of Formula I, ##STR00059## wherein A is a Met-AP2 inhibitory core; W is O or NR.sub.2; R.sub.1 and R.sub.2 are each, independently, hydrogen or alkyl; X is alkylene or substituted alkylene; n is 0 or 1; R.sub.3 and R.sub.4 are each, independently, hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl; or R.sub.3 and R.sub.4, together with the carbon atom to which they are attached, form a carbocyclic or heterocyclic group; or R.sub.3 and R.sub.4 together form an alkylene group; Z is --C(O)-- or alkylene-C(O)--; and P is a peptide comprising from 1 to about 100 amino acid residues attached at its amino terminus to Z or a group OR.sub.5 or N(R.sub.6)R.sub.7, wherein R.sub.5, R.sub.6 and R.sub.7 are each, independently, hydrogen, alkyl, substituted alkyl, azacycloalkyl or substituted azacycloalkyl; or R.sub.6 and R.sub.7, together with the nitrogen atom to which they are attached, form a substituted or unsubstituted heterocyclic ring structure; or Z is --O--, --NR.sub.8--, alkylene-O-- or alkylene-NR.sub.8--, where R.sub.8 is hydrogen or alkyl; and P is hydrogen, alkyl or a peptide consisting of from 1 to about 100 amino acid residues attached at its carboxy terminus to Z.

21. The method of claim 20, wherein said parasitic infection is malaria.

22. The method of claim 20, wherein said parasitic infection is Leishmaniasis.

23. A method of treating a thymoma in a subject, comprising administering to the subject a therapeutically effective amount of an angiogenesis inhibitor compound comprising the structure ##STR00060## wherein A is a MetAP-2 inhibitory core; W is O or NR; each R is, independently, hydrogen or alkyl; Z is --C(O)-- or -alkylene-C(O)--; P is NHR, OR or a peptide consisting of one to about one hundred amino acid residues connected at the N-terminus to Z; Q is hydrogen, linear, branched or cyclic alkyl or aryl, provided that when P is --OR, Q is not hydrogen; or Z is -alkylene-O-- or -alkylene-N(R)--; P is hydrogen or a peptide consisting of from one to about one hundred amino acid residues connected to Z at the carboxyl terminus; Q is hydrogen, linear, branched or cyclic alkyl or aryl, provided that when P is hydrogen, Q is not hydrogen; and a pharmaceutically acceptable salt thereof, thereby treating the thymoma in the subject.

24. A method of treating a thymoma in a subject, comprising administering to the subject a therapeutically effective amount of a compound comprising the structure ##STR00061## wherein A is a Met-AP2 inhibitory core; W is O or NR.sub.2; R.sub.1 and R2 are each, independently, hydrogen or alkyl; X is alkylene or substituted alkylene; n is 0 or 1; R.sub.3 and R4 are each, independently, hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl; or R.sub.3 and R.sub.4, together with the carbon atom to which they are attached, form a carbocyclic or heterocyclic group; or R.sub.3 and R.sub.4 together form an alkylene group; Z is --C(O)-- or alkylene-C(O)--; and P is a peptide comprising from 1 to about 100 amino acid residues attached at its amino terminus to Z or a group OR.sub.5 or N(R.sub.6)R.sub.7, wherein R.sub.5, R.sub.6 and R.sub.7 are each, independently, hydrogen, alkyl, substituted alkyl, azacycloalkyl or substituted azacycloalkyl; or R.sub.6 and R.sub.7, together with the nitrogen atom to which they are attached, form a substituted or unsubstituted heterocyclic ring structure; or Z is --O--, --NR.sub.8--, alkylene-O-- or alkylene-NR.sub.8--, where R.sub.8 is hydrogen or alkyl; and P is hydrogen, alkyl or a peptide consisting of from 1 to about 100 amino acid residues attached at its carboxy terminus to Z.

25. A method of treating a thymoma in a subject, comprising administering to the subject a therapeutically effective amount of a compound comprising a structure selected from the group consisting of {(3R, 4S, 5S, 6R)-5-Methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-ylox- ycarbonylamino}-3-methyl-butyric acid methyl ester; 2-{(3R, 4S, 5S, 6R)-5-Methoxy-4-[(2R, 3R)-2-methyl-3-(3 -methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-yloxycarbonylamino}-3- -methyl-butyric acid methyl ester; 2-{(3R, 4S, 5S, 6R)-5-Methoxy-4-[(2R, 3R)-2-methyl-3-(3 -methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-yloxycarbonylamino}-4- -methyl-pentanoic acid methyl ester; {(3R, 4S, 5S, 6R)-5-Methoxy-4-[(2R, 3R )-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-ylo- xycarbonylamino}-phenyl-acetic acid methyl ester; (1-Carbamoyl-2-methyl-propyl)-carbamic acid-(3R, 4S, 5S, 6R )-5-methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5 ]oct-6-yl ester; (1-Carbamoyl-2-methyl-propyl)-carbamic acid-(3R, 4S, 5S, 6R )-5-methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-butyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-yl ester; (1-Hydroxymethyl-2-methyl-propyl)-carbamic acid-(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-yl ester; 2-{(3R, 4S, 5S, 6R)-5-Methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-ylox- ycarbonylamino}-3,3-dimethyl-butyric acid methyl ester; Cyclohexyl-2-{(3R, 4S, 5S, 6R)-5-Methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-ylox- ycarbonylamino}-acetic acid methyl ester; 2-{(3R, 4S, 5S, 6R )-5-Methoxy-4-[(2R, 3R)-2-methyl-3-3 -methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-yloxycarbonylamino}-3- -methyl-pentanoic acid methyl ester; [1-(1-Carbamoyl-2-hydroxy-ethylcarbamoyl)-2-methyl-propyl]-carbamic acid-(3R, 4S, 5S, 6R )-5-methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-but-2-enyl]-oxiranyl-1-oxa-spiro[2.5]oct-6-yl ester; 2-(3-{(3R, 4S, 5S, 6R )-5-Methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]1-oxa-spiro[2.5]oct-6-yl}-u- reido)-3-methyl-butyramide; 2-{(3R, 4S, 5S, 6R)-5-Methoxy-4-[(2R, 3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[2.5]oct-6-ylox- ycarbonylamino}-3-methyl-butyric acid; N-Carbamoyl-Gly-Arg-Gly-Asp-Ser-Pro-(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-butyl)-oxiranyl]-1-oxa-spir- o[2.5]oct-6-yl ester (SEQ ID NO:31); N-Carbamoyl-Gly-Arg-Gly-Asp-Tyr-(OMe)-Arg-Glu-(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-butyl)-oxiranyl]-1-oxa-spir- o[2.5]oct-6-yl ester (SEQ ID NO:30); N-Carbamoyl-Gly-Arg-Gly-Asp-(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-butyl)-oxiranyl]-1-oxa-- spiro[2.5]oct-6-yl ester (SEQ ID NO:32); N-Carbamoyl-Gly-Arg-Gly-3-amino-3-pyridyl-propionic acid-(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa- -spiro[2.5]oct-6-yl ester (SEQ ID NO:40); N-Carbamoyl-Gly-Pro-Leu-Gly-Met-Trp-Ala-Gly-(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa- -spiro[2.5]oct-6-yl ester (SEQ ID NO:39); N-Carbamoyl-Gly-Pro-Leu-(Me)Gly-(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa- -spiro[2.5]oct-6-yl ester (SEQ ID NO:26); N-Carbamoyl-Gly-Pro-Leu-Gly-(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxirany- l]-1-oxa-spiro[2.5]oct-6-yl ester (SEQ ID NO:27); Ac-Pro-Leu-Gly-Met-Trp-Ala-(2R-{(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa- -spiro[2.5]oct-6-yloxycarbonyl}-amino-3-methyl-butanol) ester (SEQ ID NO:24); Ac-Pro-Leu-Gly-Met-GIy-(2R-{(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa- -spiro[2.5]oct-6-yloxycarbonyl}-amino-3-methyl-butanol) ester (SEQ ID NO:3 6); Met-Trp-Ala-(2R-{(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa- -spiro[2.5]oct-6-yloxycarbonyl}-amino-3-methyl-butanol) ester (SEQ ID NO:37); Met-Gly-(2R-{(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa- -spiro[2.5]oct-6-yloxycarbonyl}-amino-3-methyl-butanol) ester (SEQ ID NO :38); Ac-Pro-Leu-Gly-Met-Ala-(2R-{(3R, 4S, 5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa- -spiro[2.5]oct-6-yloxycarbonyl}-amino-3-methyl-butanol) ester (SEQ ID NO:34); {2-Methyl-1-[methyl-(1-methyl-piperidin-4-yl)-carbamoyl]-propyl}-- carbamic acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-ox- a-spiro[2.5]oct-6-yl ester; [1-(2-Dimethylamino-ethylcarbamoyl)-2-methyl-propyl]-carbamic acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[- 2.5]oct-6-yl ester; {1-[(2-Dimethylamino-ethyl)-methyl-carbamoyl]-2-methyl-propyl}-carbamic acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[- 2.5]oct-6-yl ester; [1-(3-Dimethylamino-propylcarbamoyl)-2-methyl-propyl]-carbamic acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[- 2.5]oct-6-yl ester; [1-(3-Dimethylamino-2,2-dimethyl-propylcarbamoyl)-2-methyl-propyl]-carbam- ic acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spir- o[2.5]oct-6-yl ester; [2-Methyl-1-(4-methyl-piperazine-1-carbonyl)-propyl]-carbamic acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[- 2.5]oct-6-yl ester; {2-Methyl-1-[2-(1-methyl-pyrrolidin-2-yl)-ethylcarbamoyl]-propyl}-carbami- c acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro- [2.5]oct-6-yl ester; [2-Methyl-1-(4-pyrrolidin-1-yl-piperidine-1-carbonyl)-propyl]-carbamic acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[- 2.5]oct-6-yl ester; and [1-(4-Benzyl-piperazine-1-carbonyl)-2-methyl-propyl]-carbamic acid-5-methoxy-4-[2-methyl-3-(3-methyl-but-2-enyl)-oxiranyl]-1-oxa-spiro[- 2.5]oct-6-yl ester.

26. The method of claim 1, wherein said lymphoid malignancy is a lymphoma.

27. The method of claim 26, wherein said lymphoma is a B cell lymphoma.

28. The method of claim 1, wherein said lymphoid malignancy is leukemia.

29. The method of claim 1, wherein said subject is human.

30. The method of claim 20, wherein said subject is human.

31. The method of claim 23, wherein said subject is human.

32. The method of claim 24, wherein said subject is human.

33. The method of claim 25, wherein said subject is human.

34. The method of claim 1, wherein said compound is administered to the subject using a pharmaceutically acceptable formulation.

35. The method of claim 20, wherein said compound is administered to the subject using a pharmaceutically acceptable formulation.

36. The method of claim 23, wherein said compound is administered to the subject using a pharmaceutically acceptable formulation.

37. The method of claim 24, wherein said compound is administered to the subject using a pharmaceutically acceptable formulation.

38. The method of claim 25, wherein said compound is administered to the subject using a pharmaceutically acceptable formulation.

39. The method of claim 1, wherein said compound is administered to the subject intravenously, intramuscularly or orally.

40. The method of claim 20, wherein said compound is administered to the subject intravenously, intramuscularly or orally.

41. The method of claim 23, wherein said compound is administered to the subject intravenously, intramuscularly or orally.

42. The method of claim 24, wherein said compound is administered to the subject intravenously, intramuscularly or orally.

43. The method of claim 25, wherein said compound is administered to the subject intravenously, intramuscularly or orally.

Description

BACKGROUND OF THE INVENTION

Lymphoma is a leading cause of death in the United States. Lymphoma is a type of cancer that can occur when an error occurs in the way a lymphocyte is produced, resulting in an abnormal cell. These abnormal cells can accumulate by two mechanisms: (a) they can duplicate faster than normal cells, or (b) they can live longer than normal lymphocytes. Like normal lymphocytes, the cancerous lymphocytes can grow in many parts of the body, including the lymph nodes, spleen, bone marrow, blood, or other organs. There are two main types of cancer of the lymphatic system. One is called Hodgkin's disease, while the other is called non-Hodgkin's lymphoma.

Autoimmune disorders also present a serious health issue in the United States. A progressive and maintained response by the immune system against self-components is termed autoimmunity. Normally self-tolerance mechanisms prevent the immune response from acting on self-components. However, all mechanisms have a risk of breakdown and occasionally the immune system turns on its host environment in an aggressive manner as to cause disease. This breakdown leads to the copious production of autoreactive B cells producing autoantibodies and/or autoreactive T cells leading to destructive autoimmune disease. The cellular mechanisms of autoimunity are the same as those involved in beneficial immune responses to foreign components which include antibody-dependent cell cytotoxicity, delayed-type hypersensitivity (DTH), and T-cell lympholysis.

Human autoimmune diseases can be divided into two categories: organ-specific and systemic. In organ-specific autoimmune disease, autoreactivity is directed to antigens unique to a single organ. In systemic autoimmune disease, autoreactivity is largely directed toward a broad range of antigens and involves a number of tissues. Disease in either type results from the generation of one or both autoreactive cell types (B or T cells). Autoreactive B cells lead to the generation of autoantibodies or immune complexes. Autoreactive T cells lead to the cellular DTH responses from T.sub.DTh cells or cytotoxic responses from T.sub.C cells.

Diseases caused by parasites are among the leading causes of death and disease in tropical and subtropical regions of the world. Efforts to control the invertebrate vector (carrier, such as the mosquito) of these diseases is, in many cases, difficult as a result of pesticide resistance, concerns regarding environmental damage and lack of adequate infrastructure to apply existing vector control methods. Thus, control of these diseases relies heavily on the availability of drugs. Unfortunately, most existing therapeutics are either incompletely effective or toxic to the human host. In a number of cases, even safe and effective drugs are failing as a result of the selection and spread of drug resistant variants of the parasites. This is best dramatized by the global spread of drug resistant Plasmodium falciparum, the organism responsible for the most lethal form of malaria.

Angiogenesis is the fundamental process by which new blood vessels are formed and is essential to a variety of normal body activities (such as reproduction, development and wound repair). Although the process is not completely understood, it is believed to involve a complex interplay of molecules which both stimulate and inhibit the growth of endothelial cells, the primary cells of the capillary blood vessels. Under normal conditions, these molecules appear to maintain the microvasculature in a quiescent state (i.e., one of no capillary growth) for prolonged periods which may last for as long as weeks or in some cases, decades. When necessary, however, (such as during wound repair), these same cells can undergo rapid proliferation and turnover within a 5 day period (Folkman, J. and Shing, Y., Journal of Biological Chemistry, 267(16): 10931 10934, and Folkman, J. and Klagsbrun, M. (1987) Science, 235: 442 447).

Although angiogenesis is a highly regulated process under normal conditions, many diseases (characterized as "angiogenic diseases") are driven by persistent unregulated angiogenesis. Otherwise stated, unregulated angiogenesis may either cause a particular disease directly or exacerbate an existing pathological condition. For example, ocular neovacularization has been implicated as the most common cause of blindness and dominates approximately 20 eye diseases. In certain existing conditions such as arthritis, newly formed capillary blood vessels invade the joints and destroy cartilage. In diabetes, new capillaries formed in the retina invade the vitreous, bleed, and cause blindness. Growth and metastasis of solid tumors are also angiogenesis-dependent (Folkman, J. (1986) Cancer Research 46: 467 473 and Folkman, J. (1989) Journal of the National Cancer Institute 82: 4 6). It has been shown, for example, that tumors which enlarge to greater than 2 mm, must obtain their own blood supply and do so by inducing the growth of new capillary blood vessels. Once these new blood vessels become embedded in the tumor, they provide a means for tumor cells to enter the circulation and metastasize to distant sites, such as the liver, lung or bone (Weidner, N., et al. (1991) The New England Journal of Medicine 324(1):1 8).

Fumagillin is a known compound which has been used as an antimicrobial and antiprotozoal. Its physicochemical properties and method of production are well known (U.S. Pat. No. 2,803,586 and Proc. Nat. Acad. Sci. USA (1962) 48:733 735). Fumagillin and certain types of Fumagillin analogs have also been reported to exhibit anti-angiogenic activity. However, the use of such inhibitors (e.g., TNP-470) may be limited by their rapid metabolic degradation, erratic blood levels, and by dose-limiting central nervous system (CNS) side effects.

Accordingly, there is still a need for angiogenesis inhibitors which are more potent, less neurotoxic, more stable, and/or have longer serum half-lives.

SUMMARY OF THE INVENTION

The present invention provides angiogenesis inhibitor compounds which comprise a core, e.g., a Fumagillin core, that is believed to inhibit methionine aminopeptidase 2 (MetAP-2), coupled to a peptide. The present invention is based, at least in part, on the discovery that coupling the MetAP-2 inhibitory core to an amino acid residue or an amino acid derivative prevents the metabolic degradation of the angiogenesis inhibitor compound to ensure a superior pharmacokinetic profile and limits CNS side effects by altering the ability of the angiogenesis inhibitor compound to cross the blood brain barrier. The present invention is also based, at least in part, on the discovery that coupling the MetAP-2 inhibitory core to a peptide comprising a site-directed sequence allows for a cell specific delivery of the angiogenesis inhibitor compound and limits the toxicity of the angiogenesis inhibitor compound.

In one aspect the present invention provides a method for treating a subject (e.g., a mammal, such as a human) suffering from a lymphoid malignancy. The method includes administering to a subject an effective amount of a MetAP-2 inhibitor, thereby treating a subject suffering from a lymphoid malignancy. Lymphoid malignancies which can be treated with a MetAP-2 inhibitor include lymphoid leukemias, such as chronic lymphoid leukemia and acute lymphoid leukemia, and lymphomas, such as T cell lymphoma and B cell lymphoma.

In a preferred embodiment, the method further includes administering to the subject a second therapy suitable for treating a subject suffering from lymphoid malignancy. The second therapy may be administered to the subject subsequent to, simultaneously or prior to administration of the MetAP-2 inhibitor to the subject. The second therapy may include administration of a chemotherapeutic regimen or a vaccine to the subject.

Accordingly, the present invention provides compounds of Formula I,

##STR00001##

In Formula I, A is a MetAP-2 inhibitory core,W is O or NR2, and R.sub.1 and R.sub.2 are each, independently, hydrogen or alkyl; X is alkylene or substituted alkylene, preferably linear C.sub.1 C.sub.6-alkylene; n is 0 or 1; R.sub.3 and R.sub.4 are each, independently, hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aryl or arylalkyl or substituted or unsubstituted heteroaryl or heteroalkyl. R.sub.3 and R.sub.4 can also, together with the carbon atom to which they are attached, form a carbocyclic or heterocyclic group; or R.sub.1 and R.sub.4 together can form an alkylene group; Z is --C(O)--, alkylene-C(O)-- or alkylene; and P is a peptide comprising from 1 to about 100 amino acid residues attached at its amino terminus to Z or a group OR.sub.5 or N(R.sub.6)R.sub.7, wherein R.sub.5, R.sub.6 and R.sub.7 are each, independently, hydrogen, alkyl, substituted alkyl, azacycloalkyl or substituted azacycloalkyl. R.sub.6 and R.sub.7 can also form, together with the nitrogen atom to which they are attached, a substituted or unsubstituted heterocyclic ring structure.

In another embodiment of the compounds of Formula I, W, X, n, R.sub.1, R.sub.3 and R.sub.4 have the meanings given above for these variables; Z is --O--, --NR.sub.8--, alkylene-O-- or alkylene-NR.sub.8--, where R.sub.8 is hydrogen or alkyl; and P is hydrogen, alkyl, preferably normal or branched C.sub.1 C.sub.4-alkyl or a peptide consisting of from 1 to about 100 amino acid residues attached at its carboxy terminus to Z.

In compounds of Formula I, when any of R.sub.1 R.sub.8 is an alkyl group, preferred alkyl groups are substituted or unsubstituted normal, branched or cyclic C.sub.1 C.sub.6 alkyl groups. Particularly preferred alkyl groups are normal or branched C.sub.1 C.sub.4 alkyl groups. A substituted alkyl group includes at least one non-hydrogen substituent, such as an amino group, an alkylamino group or a dialkylamino group; a halogen, such as a fluoro, chloro, bromo or iodo substituent; or hydroxyl.

When at least one of R.sub.3 and R.sub.4 is a substituted or unsubstituted aryl or heteroaryl group, preferred groups include substituted and unsubstituted phenyl, naphthyl, indolyl, imidazoly and pyridyl. When at least one of R.sub.3 and R.sub.4 is substituted or unsubstituted arylalkyl or heteroarylalkyl, preferred groups include substituted and unsubstituted benzyl, naphthylmethyl, indolylmethyl, imidazolylmethyl and pyridylmethyl groups. Preferred substituents on aryl, heteroaryl, arylalkyl and heteroarylalkyl groups are independently selected from the group consisting of amino, alkyl-substituted amino, halogens, such as fluoro, chloro, bromo and iodo; hydroxyl groups and alkyl groups, preferably normal or branched C.sub.1 C.sub.6-alkyl groups, most preferably methyl groups. X is preferably linear C.sub.1 C.sub.6-alkylene, more preferably C.sub.1 C.sub.4-alkylene and most preferably methylene or ethylene. When Z is alkylene-C(O)--, alkylene-O-- or alkylene-NR.sub.8, the alkylene group is preferably linear C.sub.1 C.sub.6-alkylene, more preferably C.sub.1 C.sub.4-alkylene and most preferably methylene or ethylene.

R.sub.6 and R.sub.7, in addition to alkyl, substituted alkyl or hydrogen, can each also independently be a substituted or unsubstituted azacycloalkyl group or a substituted or unsubstituted azacycloalkylalkyl group. Suitable substituted azacycloalkyl groups include azacycloalkyl groups which have an N-alkyl substituent, preferably an N--C.sub.1 C.sub.4-alkyl substituent and more preferably an N-methyl substituent. R.sub.6 and R.sub.7 can also, together with the nitrogen atom to which they are attached, form a heterocyclic ring system, such as a substituted or unsubstituted five or six-membered aza- or diazacycloalkyl group. Preferably, the diazacycloalkyl group includes an N-alkyl substituent, such as an N--C.sub.1 C.sub.4-alkyl substituent or, more preferably, an N-methyl substituent.

In particularly preferred embodiments, --N(R.sub.6)R.sub.7 is NH.sub.2 or one of the groups shown below:

##STR00002##

Preferably, the compounds of Formula I do not include compounds wherein Z is --O--, P is hydrogen, R.sub.3 and R.sub.4 are both hydrogen, n is 1 and X is methylene. Preferably, the compounds of Formula I further do not include compounds wherein Z is methylene-O--, R.sub.3 and R.sub.4 are both hydrogen, and n is 0.

In another aspect, the present invention is directed to angiogenesis inhibitor compounds of Formula XV,

##STR00003## where A is a MetAP-2 inhibitory core and W is O or NR. In one embodiment, Z is --C(O)-- or -alkylene-C(O)-- and P is NHR, OR or a peptide consisting of one to about one hundred amino acid residues connected at the N-terminus to Z. In this embodiment, Q is hydrogen, linear, branched or cyclic alkyl or aryl, provided that when P is --OR, Q is not hydrogen.

In another embodiment, Z is -alkylene-O-- or -alkylene-N(R)-- and P is hydrogen or a peptide consisting of from one to about one hundred amino acid residues connected to Z at the carboxyl terminus. In this embodiment, Q is hydrogen, linear, branched or cyclic alkyl or aryl, provided that when P is hydrogen, Q is not hydrogen.

In the angiogenesis inhibitor compounds of Formula XV, each R is, independently, hydrogen or alkyl.

In another aspect, the invention features pharmaceutical compositions comprising the angiogenesis inhibitor compounds of Formula I or XV and a pharmaceutically acceptable carrier.

In yet another aspect, the invention features a method of treating an angiogenic disease, e.g., cancer (such as lung cancer, brain cancer, kidney cancer, colon cancer, liver cancer, pancreatic cancer, stomach cancer, prostate cancer, breast cancer, ovarian cancer, cervical cancer, melanoma, and metastatic versions of any of the preceding cancers), in a subject. The method includes administering to the subject a therapeutically effective amount of one or more angiogenesis inhibitor compounds of Formula I or XV.

In one embodiment, the present invention provides a method of treating a subject suffering from a parasitic infection, such as an infection by Plasmodium species, such as Plasmodium falciparum, or an infection by Leishmania species, such as Leishmania donavani. The method comprises the step of administering to the subject a therapeutically effective amount of a compound of the invention. The subject can be an individual who is suffering from, or susceptible to, infection by a parasitic organism. In a preferred embodiment, the subject suffers from malaria or Leishmaniasis.

The invention further provides a method of treating a subject suffering from a lymphoid malignancy. The method comprises the step of administering to the subject a therapeutically effective amount of a compound of the invention. Suitable lymphoid malignancies which can be treated with a compound of the invention include lymphoid leukemias, such as chronic lymphoid leukemia and acute lymphoid leukemia, and lymphomas, such as Non-Hodgkin's lymphoma, including T cell lymphoma and B cell lymphoma.

In a further embodiment, the invention provides a method of treating a subject suffering from an autoimmune disorder, comprising the step of administering to the subject a therapeutically effective amount of a compound of the invention. The autoimmune disorder can be, for example, rheumatoid arthritis, lupus erythematosus, psoriasis, multiple sclerosis, myasthenia gravis, vasculitis, or diabetes mellitus.

Other features and advantages of the invention will be apparent from the following detailed description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a series of graphs depicting the inhibition of SR cell proliferation in culture following 3 or 6 days of exposure to Compound 5 (representative data).

FIG. 2 is a graph depicting tumor volumes of SR lymphoma tumor-bearing mice treated with Compound 5.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides compounds useful as angiogenesis inhibitors and methods for using these compounds in the treatment of angiogenic diseases. Without intending to be limited by theory, it is believed that the angiogenesis inhibitor compounds of the invention inhibit angiogenesis by inhibiting methionine aminopeptidase 2 (MetAP-2), an enzyme which cleaves the N-terminal methionine residue of newly synthesized proteins to produce the active form of the protein. At the same time, the presence of a peptide in the angiogenesis inhibitor compounds of the invention prevents the metabolic degradation of the angiogenesis inhibitor compounds and ensures a superior pharmacokinetic profile. The presence of the peptide in the angiogenesis inhibitor compounds of the invention also alters the ability of the angiogenesis inhibitor compound to cross the blood brain barrier to, for example, limit CNS side effects (such as CNS toxicity). The presence of peptides comprising a site-directed sequence in the-angiogenesis inhibitor compounds of the invention allows for a site-specific delivery of the angiogenesis inhibitor compounds and, thus, limits the toxicity of the angiogenesis inhibitor compounds.

The angiogenesis inhibitor compounds of the invention comprise a MetAP-2 inhibitory core and a peptide attached, directly or indirectly, thereto. In one embodiment, the invention provides angiogenesis inhibitor compounds of Formula I

##STR00004##

In Formula I, A is a MetAP-2 inhibitory core, W is O or NR.sub.2, and R.sub.1 and R.sub.2 are each, independently, hydrogen or alkyl; X is alkylene or substituted alkylene, preferably linear C.sub.1 C.sub.6-alkylene; n is 0 or 1; R.sub.3 and R.sub.4 are each, independently, hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aryl or arylalkyl or substituted or unsubstituted heteroaryl or heteroalkyl. R.sub.3 and R.sub.4 can also, together with the carbon atom to which they are attached, form a carbocyclic or heterocyclic group; or R.sub.1 and R.sub.4 together can form an alkylene group; Z is --C(O)--, alkylene-C(O)-- or alkylene; and P is a peptide comprising from 1 to about 100 amino acid residues attached at its amino terminus to Z or a group OR.sub.5 or N(R.sub.6)R.sub.7, wherein R.sub.5, R.sub.6 and R.sub.7 are each, independently, hydrogen, alkyl, substituted alkyl, azacycloalkyl or substituted azacycloalkyl. R.sub.6 and R.sub.7 can also form, together with the nitrogen atom to which they are attached, a substituted or unsubstituted heterocyclic ring structure.

In another embodiment of the compounds of Formula I, W, X, n, R.sub.1, R.sub.3 and R.sub.4 have the meanings given above for these variables; Z is --O--, --NR.sub.8--, alkylene-O-- or alkylene-NR.sub.8--, where R.sub.8 is hydrogen or alkyl; and P is hydrogen, alkyl, preferably normal or branched C.sub.1 C.sub.4-alkyl or a peptide consisting of from 1 to about 100 amino acid residues attached at its carboxy terminus to Z.

In compounds of Formula I, when any of R.sub.1 R.sub.8 is an alkyl group, preferred alkyl groups are substituted or unsubstituted normal, branched or cyclic C.sub.1 C.sub.6 alkyl groups. Particularly preferred alkyl groups are normal or branched C.sub.1 C.sub.4 alkyl groups. A substituted alkyl group includes at least one non-hydrogen substituent, such as an amino group, an alkylamino group or a dialkylamino group; a halogen, such as a fluoro, chloro, bromo or iodo substituent; or hydroxyl.

When at least one of R.sub.3 and R.sub.4 is a substituted or unsubstituted aryl or heteroaryl group, preferred groups include substituted and unsubstituted phenyl, naphthyl, indolyl, imidazolyl and pyridyl. When at least one of R.sub.3 and R.sub.4 is substituted or unsubstituted arylalkyl or heteroarylalkyl, preferred groups include substituted and unsubstituted benzyl, naphthylmethyl, indolylmethyl, imidazolylmethyl and pyridylmethyl groups. Preferred substituents on aryl, heteroaryl, arylalkyl and heteroarylalkyl groups are independently selected from the group consisting of amino, alkyl-substituted amino, halogens, such as fluoro, chloro, bromo and iodo; hydroxyl groups and alkyl groups, preferably normal or branched C.sub.1 C.sub.6-alkyl groups, most preferably methyl groups. X is preferably linear C.sub.1 C.sub.6-alkylene, more preferably C.sub.1 C.sub.4-alkylene and most preferably methylene or ethylene. When Z is alkylene-C(O)--, alkylene-O-- or alkylene-NR.sub.8, the alkylene group is preferably linear C.sub.1 C.sub.6-alkylene, more preferably C.sub.1 C.sub.4-alkylene and most preferably methylene or ethylene.

R.sub.6 and R.sub.7, in addition to alkyl, substituted alkyl or hydrogen, can each also independently be a substituted or unsubstituted azacycloalkyl group or a substituted or unsubstituted azacycloalkylalkyl group. Suitable substituted azacycloalkyl groups include azacycloalkyl groups which have an N-alkyl substituent, preferably an N--C.sub.1 C.sub.4-alkyl substituent and more preferably an N-methyl substituent. R.sub.6 and R.sub.7 can also, together with the nitrogen atom to which they are attached, form a heterocyclic ring system, such as a substituted or unsubstituted five or six-membered aza- or diazacycloalkyl group. Preferably, the diazacycloalkyl group includes an N-alkyl substituent, such as an N--C.sub.1 C.sub.4-alkyl substituent or, more preferably, an N-methyl substituent.

In particularly preferred embodiments, --N(R.sub.6)R.sub.7 is NH.sub.2 or one of the groups shown below:

##STR00005##

Preferably, the compounds of Formula I do not include compounds wherein Z is --O--, P is hydrogen, R.sub.3 and R.sub.4 are both hydrogen, n is 1 and X is methylene. Preferably, the compounds of Formula I further do not include compounds wherein Z is methylene-O--, R.sub.3 and R.sub.4 are both hydrogen, and n is 0.

In another embodiment, the invention provides angiogenesis inhibitor compounds of Formula XV,

##STR00006## where A is a MetAP-2 inhibitory core and W is O or NR. In one embodiment, Z is --C(O)-- or -alkylene-C(O)-- and P is NHR, OR or a peptide consisting of one to about one hundred amino acid residues connected at the N-terminus to Z. In this embodiment, Q is hydrogen, linear, branched or cyclic alkyl or aryl, provided that when P is --OR, Q is not hydrogen. Z is preferably --C(O)-- or C.sub.1 C.sub.4-alkylene-C(O)--, and, more preferably, --C(O)-- or C.sub.1 C.sub.2-alkylene-C(O)--. Q is preferably linear, branched or cyclic C.sub.1 C.sub.6-alkyl, phenyl or naphthyl. More preferably, Q is isopropyl, phenyl or cyclohexyl.

In another embodiment, Z is -alkylene-O-- or -alkylene-N(R)--, where alkylene is, preferably, C.sub.1 C.sub.6-alkylene, more preferably C.sub.1 C.sub.4-alkylene and, most preferably, C.sub.1 C.sub.2-alkylene. P is hydrogen or a peptide consisting of from one to about one hundred amino acid residues connected to Z at the carboxyl terminus. In this embodiment, Q is hydrogen, linear, branched or cyclic alkyl or aryl, provided that when P is hydrogen, Q is not hydrogen. Q is preferably linear, branched or cyclic C.sub.1 C.sub.6-alkyl , phenyl or naphthyl. More preferably, Q is isopropyl, phenyl or cyclohexyl.

In the compounds of Formula XV, each R is, independently, hydrogen or alkyl. In one embodiment, each R is, independently, hydrogen or linear, branched or cyclic C.sub.1 C.sub.6-alkyl. Preferably, each R is, independently, hydrogen or linear or branched C.sub.1 C.sub.4-alkyl. More preferably, each R is, independently, hydrogen or methyl. In the most preferred embodiments, each R is hydrogen.

In Formulas I and XV, A is a MetAP-2 inhibitory core. As used herein, a "MetAP-2 inhibitory core" includes a moiety able to inhibit the activity of methionine aminopeptidase 2 (MetAP-2), e.g., the ability of MetAP-2 to cleave the N-terminal methionine residue of newly synthesized proteins to produce the active form of the protein. Preferred MetAP-2 inhibitory cores are Fumagillin derived structures.

Suitable MetAP-2 inhibitory cores include the cores of Formula II,

##STR00007## where R.sup.1 is hydrogen or alkoxy, preferably C.sub.1 C.sub.4-alkoxy and more preferably, methoxy. R.sup.2 is hydrogen or hydroxy; and R.sup.3 is hydrogen or alkyl, preferably C.sub.1 C.sub.4-alkyl and more preferably, hydrogen. D is linear or branched alkyl, preferably C.sub.1 C.sub.6-alkyl; arylalkyl, preferably aryl-C.sub.1 C.sub.4-alkyl and more preferably phenyl-C.sub.1 C.sub.4-alkyl; or D is of the structure

##STR00008## where the dashed line represents a single bond or a double bond.

A can also be a MetAP-2 inhibitory core of Formula III,

##STR00009## ##STR00010## Where R.sup.1, R.sup.2, R.sup.3 and D have the meanings given above for Formula II, and X is a leaving group, such as a halogen.

Examples of suitable MetAP-2 inhibitory cores include, but are not limited to, the following.

In each of Formulas IV X, the indicated valence on the ring carbon is the point of attachment of the structural variable W, as set forth in Formulas I XV. In Formula IX, p is an integer from 0 to 10, preferably 1 4. In Formulas IV, V and VI IX, R.sub.1 is hydrogen or C.sub.1 C.sub.4-alkoxy, preferably methoxy. In Formulas IV and V, the dashed line indicates that the bond can be a double bond or a single bond. In Formula V, X represents a leaving group, such as a thioalkoxy group, a thioaryloxy group, a halogen or a dialkylsulfinium group. In Formulas IV and V, R.sub.2 is H, OH, amino, C.sub.1 C.sub.4-alkylamino or di(C.sub.1 C.sub.4-alkyl)amino), preferably H. In formulas in which the stereochemistry of a particular stereocenter is not indicated, that stereocenter can have either of the possible stereochemistries, consistent with the ability of the angiogenesis inhibitor compound to inhibit the activity of MetAP-2.

In particularly preferred embodiments, A is the MetAP-2 inhibitory core of Formula X below.

##STR00011##

As used herein, the terms "P" and "peptide" include compounds comprising from 1 to about 100 amino acid residues (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acid residues). In preferred embodiments, the peptide includes compounds comprising less than about 90, 80, 70, 60, 50, 40, 30, 20, or 10 amino acid residues, preferably about 1 10, 1 20, 1 30, 1 40, 1 50, 1 60, 1 70, 1 80, or 1 90 amino acid residues. The peptides may be natural or synthetically made. The amino acid residues are preferably .alpha.-amino acid residues. For example, the amino acid residues can be independently selected from among the twenty naturally occurring amino acid residues, the D-enantiomers of the twenty natural amino acid residues, and may also be non-natural amino acid residues (e.g., norleucine, norvaline, phenylglycine, .beta.-alanine, or a peptide mimetic such as 3-amino-methylbenzoic acid). In one embodiment, the amino acid residues are independently selected from residues of Formula XI, Formula XII, and Formula XIII.

##STR00012##

In Formula XI, X.sub.1 is hydrogen, a side chain of one of the twenty naturally-occurring amino acid residues, a linear, branched or cyclic C.sub.1 C.sub.8-alkyl group, an aryl group, such as a phenyl or naphthyl group, an aryl-C.sub.1 C.sub.4-alkyl group, a heteroaryl group, such as a pyridyl, thienyl, pyrrolyl, or furyl group, or a heteroaryl-C.sub.1 C.sub.4-alkyl group; and X.sub.2 is hydrogen a linear, branched or cyclic C.sub.1 C.sub.8-alkyl group, an aryl group, such as a phenyl or naphthyl group, an aryl-C.sub.1 C.sub.4-alkyl group or a heteroaryl group as described above for X.sub.1. Preferably, X.sub.2 is hydrogen. In Formula XII, Y is methylene, oxygen, sulfur or NH, and a and b are each, independently, 0 4, provided that the sum of a and b is between 1 and 4. Formulas XI and XII encompass .alpha.-amino acid residues having either a D or an L stereochemistry at the alpha carbon atom. One or more of the amino acid residues can also be an amino acid residue other than an .alpha.-amino acid residue, such as a .beta.-, .gamma.- or .epsilon.-amino acid residue. Suitable examples of such amino acid residues are of Formula XIII, wherein q is an integer of from 2 to about 6, and each X.sub.1 and X.sub.2 independently have the meanings given above for these variables in Formula XI.

In a preferred embodiment, the peptide used in the angiogenesis inhibitor compounds of the invention may include a site-directed sequence in order to increase the specificity of binding of the angiogenesis inhibitor compound to a cell surface of interest. As used herein, the term "site-directed sequence" is intended to include any amino acid sequence (e.g., comprised of natural or non natural amino acid residues) which serves to limit exposure of the angiogenesis inhibitor compound to the periphery and/or which serves to direct the angiogenesis inhibitor compound to a site of interest, e.g., a site of angiogenesis or aberrant cellular proliferation.

The peptide contained within the angiogenesis inhibitor compounds of the invention may include a peptide cleavage site for an enzyme which is expressed at sites of angiogenesis or aberrant cell proliferation, allowing tissue-selective delivery of a cell-permeable active angiogenesis inhibitor compound or fragment thereof (e.g., a fragment containing the MetAP-2 inhibitory core of the angiogenesis inhibitor compound). The peptide may also include a sequence which is a ligand for a cell surface receptor which is expressed at a site of angiogenesis or aberrant cell proliferation, thereby targeting angiogenesis inhibitor compounds to a cell surface of interest. For example, a peptide contained within the angiogenesis inhibitor compounds of the invention can include a cleavage site for a matrix metalloproteinase, or an integrin binding RGD (Arg-Gly-Asp) sequence, or a combination of both an enzyme "cleavage" sequence and a cell surface "ligand" which serve to target the angiogenesis inhibitor compound to the membrane of an endothelial cell. However, the selection of a peptide sequence must be such that the active angiogenesis inhibitor compound is available to be delivered to the cells in which MetAP-2 inhibition is desired.

For example, a sequence that is cleaved by a matrix matalloproteinase produces a product that contains the MetAP-2 inhibitory core, a coupling group, and a peptide fragment. Sequences are selected so that the active angiogenesis inhibitor compound, e.g., the active angiogenesis inhibitor compound generated by the matrix matalloproteinase cleavage, is cell permeable. Preferably, the active angiogenesis inhibitor compound does not contain a free acid after the cleavage.

In one embodiment, the peptide includes a cleavage site for a matrix metalloprotease, such as matrix metalloprotease-2 (MMP-2), MMP-1, MMP-3, MMP-7, MMP-8, MMP-9, MMP-12, MMP-13 or MMP-26. Preferably, the peptide includes a cleavage site for MMP-2 or MMP-9. For example, the peptide can comprise the sequence -Pro-Leu-Gly-Xaa-(SEQ ID NO:1), where Xaa is any naturally occurring amino acid residue consistent with matrix metalloprotease (MMP) cleavage at the Gly-Xaa bond. Xaa is preferably a hydrophobic amino acid residue, such as tryptophan, phenylalanine, methionine, leucine, isoleucine, proline, and valine.

Other suitable sequences include sequences comprising one or more of Pro-Cha-Gly-Cys(Me)-His (SEQ ID NO:2); Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg (SEQ ID NO:3); Pro-Gln-Gly-Ile-Ala-Gly-Trp (SEQ ID NO:4); Pro-Leu-Gly-Cys(Me)-His-Ala-D-Arg (SEQ ID NO:5); Pro-Leu-Gly-Met-Trp-Ser-Arg (SEQ ID NO:35); Pro-Leu-Gly-Leu-Trp-Ala-D-Arg (SEQ ID NO:6); Pro-Leu-Ala-Leu-Trp-Ala-Arg (SEQ ID NO:7); Pro-Leu-Ala-Leu-Trp-Ala-Arg (SEQ ID NO:8); Pro-Leu-Ala-Tyr-Trp-Ala-Arg (SEQ ID NO:9); Pro-Tyr-Ala-Tyr-Trp-Met-Arg (SEQ ID NO:10); Pro-Cha-Gly-Nva-His-Ala (SEQ ID NO:11); Pro-Leu-Ala-Nva (SEQ ID NO:12); Pro-Leu-Gly-Leu (SEQ ID NO:13); Pro-Leu-Gly-Ala (SEQ ID NO:14); Arg-Pro-Leu-Ala-Leu-Trp-Arg-Ser (SEQ ID NO:15); Pro-Cha-Ala-Abu-Cys(Me)-His-Ala (SEQ ID NO:16); Pro-Cha-Ala-Gly-Cys(Me)-His-Ala (SEQ ID NO:17); Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu (SEQ ID NO:18); Pro-Lys-Pro-Leu-Ala-Leu (SEQ ID NO:19); Arg-Pro-Lys-Pro-Tyr-Ala-Nva-Trp-Met (SEQ ID NO:20); Arg-Pro-Lys-Pro-Val-Glu-Nva-Trp-Arg (SEQ ID NO:21); Arg-Pro-Lys-Pro-Val-Glu-Nva-Trp-Arg (SEQ ID NO:22); and Arg-Pro-Lys-Pro-Leu-Ala-Nva-Trp (SEQ ID NO:23). These sequences identify the natural amino acid residues using the customary three-letter abbreviations; the following abbreviations represent the indicated non-natural amino acids: Abu=L-a-aminobutyryl; Cha=L-cyclohexylalanine; Nva=L-norvaline.

In certain embodiments, P is an amino acid sequence selected from the group consisting of Ac-Pro-Leu-Gly-Met-Trp-Ala (SEQ ID NO:24); Gly-Pro-Leu-Gly-Met-His-Ala-Gly (SEQ ID NO:25); Gly-Pro-Leu-(Me)Gly (SEQ ID NO:26); Gly-Pro-Leu-Gly (SEQ ID NO:27); Gly-Met-Gly-Leu-Pro (SEQ ID NO:28); Ala-Met-Gly-Ile-Pro (SEQ ID NO:29); Gly-Arg-Gly-Asp-(O-Me-Tyr)-Arg-Glu (SEQ ID NO:30); Gly-Arg-Gly-Asp-Ser-Pro (SEQ ID NO:31); Gly-Arg-Gly-Asp (SEQ ID NO:32); Asp-Gly-Arg; Ac-Pro-Leu-Gly-Met-Ala (SEQ ID NO:34); Ac-Arg-Gly-Asp-Ser-Pro-Leu-Gly-Met-Trp-Ala (SEQ ID NO:33); Ac-Pro-Leu-Gly-Met-Gly (SEQ ID NO:36); Met-Trp-Ala (SEQ ID NO:37); Met-Gly (SEQ ID NO:38); Gly-Pro-Leu-Gly-Met-Trp-Ala-Gly (SEQ ID NO:39); and Gly-Gly-Arg-(3-amino-3-pyridylpropionic acid) (SEQ ID NO:40). (Ac in the foregoing sequences represents an Acetyl group).

The peptide can be attached to the MetAP-2 inhibitory core at either its N-terminus or C-terminus. When the peptide is attached to the MetAP-2 inhibitory core at its C-terminus, the N-terminus of the peptide can be --NR.sub.2R.sub.3, where R.sub.2 is hydrogen, alkyl or arylalkyl and R.sub.3 is hydrogen, alkyl, arylalkyl or acyl. When the peptide is attached to the MetAP-2 inhibitory core at its N-terminus, the C-terminus can be --C(O)R.sub.4, where R.sub.4 is --OH, --O-alkyl, --O-arylalkyl, or --NR.sub.2R.sub.3, where R.sub.2 is hydrogen, alkyl or arylalkyl and R.sub.3 is hydrogen, alkyl, arylalkyl or acyl. In this embodiment, the C-terminal residue can also be present in a reduced form, such as the corresponding primary alcohol.

The present invention also includes pharmaceutically acceptable salts of the angiogenesis inhibitor compounds of the invention. A "pharmaceutically acceptable salt" includes a salt that retains the desired biological activity of the parent angiogenesis inhibitor compound and does not impart any undesired toxicological effects. Examples of such salts are salts of acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosporic acid, nitric acid, and the like; acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, benzoic acid, pamoic acid, alginic acid, methanesulfonic acid, naphthalenesulfonic acid, and the like. Also included are salts of cations such as sodium, po