Triazole-crosslinked and thioether-crosslinked peptidomimetic macrocycles

1. A peptidomimetic macrocycle comprising an amino acid sequence which is at least 90% identical to an amino acid sequence of SEQ ID NO. 455, wherein the peptidomimetic macrocycle has the formula:

an isomer or pharmaceutically acceptable salt thereof

wherein:

each A, C, D, and E is independently an amino acid;

each B is independently an amino acid,

[—NH-L 3 -CO—], [—NH-L 3 -SO 2 —], or [—NH-L 3 -];

each R 1 and R 2 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-; or at least one of R 1 and R 2 forms a macrocycle-forming linker L′ connected to the alpha position of one of said D or E amino acids;

each L and L′ is independently a macrocycle-forming linker of the formula

each L 1 , L 2 and L 3 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R 4 —K—R 4 —] n , each being optionally substituted with R 5 ;

each R 3 is independently hydrogen, alkyl, alkenyl, alkynyl, arylalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R 5 ;

each R 4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;

each K is independently O, S, SO, SO 2 , CO, CO 2 , or CONR 3 ;

each R 5 is independently halogen, alkyl, —OR 6 , —N(R 6 ) 2 , —SR 6 , —SOR 6 , —SO 2 R 6 , —CO 2 R 6 , a fluorescent moiety, a radioisotope or a therapeutic agent;

each R 6 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heterocycloalkyl, a fluorescent moiety, a radioisotope or a therapeutic agent;

each R 7 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R 5 , or part of a cyclic structure with a D residue;

each R 8 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R 5 , or part of a cyclic structure with an E residue;

v is an integer from 1-1000;

w is an integer from 3-1000;

u is an integer from 1-10;

each x, y and z is independently an integer from 0-10; and

n is an integer from 1-5.

2. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which has improved binding affinity to MDM2 or MDMX relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

3. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which has a reduced ratio of binding affinities to MDMX versus MDM2 relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

4. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which has improved in vitro anti-tumor efficacy against p53 positive tumor cell lines relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

5. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which shows improved in vitro induction of apoptosis in p53 positive tumor cell lines relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

6. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which has an improved in vitro anti-tumor efficacy ratio for p53 positive versus p53 negative or mutant tumor cell lines relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

7. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which has improved in vivo anti-tumor efficacy against p53 positive tumors relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

8. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which has improved in vivo induction of apoptosis in p53 positive tumors relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

9. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which has improved cell permeability relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

10. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , which has improved solubility relative to a corresponding peptidomimetic macrocycle with a w of 0, 1 or 2.

11. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein each E is independently an amino acid selected from Ala (alanine), D-Ala (D-alanine), Aib (α-aminoisobutyric acid), Sar (N-methyl glycine), and Ser (serine).

12. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein [D] v is -Leu 1 -Thr 2 -Phe 3 .

13. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein w is 3-10.

14. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 13 , wherein w is 3-6.

15. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 13 , wherein w is 6-10.

16. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 15 , wherein w is 6.

17. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein v is 1-10.

18. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 17 , wherein v is 2-10.

19. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 18 , wherein v is 2-5.

20. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 18 , wherein v is 2.

21. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein each E is independently Ser or Ala or an analog of Ser or Ala.

22. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , comprising at least one amino acid which is an amino acid analog.

23. A method of treating cancer in a subject comprising administering to the subject a peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 .

24. A method of modulating the activity of p53 and/or MDM2 and/or MDMX in a subject comprising administering to the subject a peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 .

25. A method of antagonizing the interaction between p53 and MDM2 and/or between p53 and MDMX proteins in a subject comprising administering to the subject a peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 .

26. The peptidomimetic macrocycle of claim 1 , which is at least 95% identical to an amino acid sequence of SEQ ID NO. 455, an isomer or pharmaceutically acceptable salt thereof.

27. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein v is an integer from 1-500.

28. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein v is an integer from 1-200.

29. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein v is an integer from 1-100.

30. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein v is an integer from 1-50.

31. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein v is an integer from 1-30.

32. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein the sum of x+y+z is 2, 3, or 6.

33. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein w is an integer from 3-500.

34. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein w is an integer from 3-200.

35. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein w is an integer from 3-100.

36. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein w is an integer from 3-50.

37. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 or 32 , wherein w is an integer from 3-30.

38. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 or 31 , wherein w is an integer from 3-20.

39. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claims 1 or 32 , wherein v is an integer from 1-20.

40. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 , wherein u is an integer from 1-5.

41. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 or 31 , wherein u is an integer from 1-3.

42. The peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of claim 1 or 32 , wherein u is an integer from 1-2.

43. The peptidomimetic macrocycle of claim 1 , consisting of an amino acid sequence of SEQ ID NO. 455, an isomer or pharmaceutically acceptable salt thereof.

44. A pharmaceutical composition comprising the peptidomimetic macrocycle, isomer or pharmaceutically acceptable salt of any one of claims 1 , 26 , and 43 .