Peptidomimetic macrocycles

1. A method of treating a condition in a subject in need thereof, the method comprising administering to the subject a therapeutically-effective amount of a peptidomimetic macrocycle, wherein the peptidomimetic macrocycle comprises an amino acid sequence which is at least about 60% identical to GHRH 1-29, wherein the peptidomimetic macrocycle comprises at least two macrocycle-forming linkers, wherein the first of the at least two macrocycle-forming linkers connects a first amino acid to a second amino acid, and the second of the at least two macrocycle-forming linkers connects a third amino acid to a fourth amino acid, wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid, wherein the peptidomimetic macrocycle comprises a helix.

2. The method of claim 1 , wherein the amino acid sequence of the peptidomimetic macrocycle is at least about 60% identical to an amino acid sequence chosen from the group consisting of SEQ ID NO. 2-180.

3. The method of claim 1 , wherein the amino acid sequence of said peptidomimetic macrocycle is at least about 80% identical to an amino acid sequence chosen from the group consisting of SEQ ID NO. 2-180.

4. The method of claim 1 , wherein the amino acid sequence of said peptidomimetic macrocycle is at least about 90% identical to an amino acid sequence chosen from the group consisting of SEQ ID NO. 2-180.

5. The method of claim 1 , wherein the amino acid sequence of said peptidomimetic macrocycle is chosen from the group consisting of SEQ ID NO. 2-180.

6. The method of claim 1 , wherein the peptidomimetic macrocycle comprises an α-helix.

7. The method of claim 1 , wherein the condition is a cachexia.

8. The method of claim 1 , wherein the condition is a lipodystrophy.

9. The method of claim 1 , wherein the condition is a growth hormone disorder.

10. The method of claim 1 , wherein each of the at least two macrocycle-forming linkers connects a pair of amino acids corresponding to one of the following locations of amino acids: 4 and 8; 5 and 12; 8 and 12; 8 and 15; 9 and 16; 12 and 16; 12 and 19; 15 and 22; 18 and 25; 21 and 25; 21 and 28; 22 and 29; or 25 and 29 of GHRH 1-29.

11. The method of claim 1 , wherein each of the at least two macrocycle-forming linkers connects a pair of amino acids corresponding to one of the following locations of amino acids: 4 and 8; 5 and 12; 12 and 19; 15 and 22; 18 and 25; 21 and 25; or 21 and 28 of GHRH 1-29.

12. The method of claim 1 , wherein the first macrocycle-forming linker connects a pair of amino acids corresponding to one of the following locations of amino acids: 4 and 8; 5 and 12; 8 and 12; 8 and 15; 9 and 16; 12 and 16; or 12 and 19 of GHRH 1-29; and the second macrocycle-forming linker connects a pair of amino acids corresponding to one of the following locations of amino acid 15 and 22; 18 and 25; 21 and 25; 21 and 28; 22 and 29; or 25 and 29 of GHRH 1-29.

13. The method of claim 1 , wherein the first macrocycle-forming linker connects a pair of amino acids corresponding to one of the following locations of amino acids: 4 and 8; 5 and 12; 8 and 12; or 12 and 19 of GHRH 1-29; and the second macrocycle-forming linker connects a pair of amino acids corresponding to one of the following locations of amino acid 15 and 22; 18 and 25; 21 and 25; or 21 and 28 of GHRH 1-29.

14. The method of claim 1 , wherein the first macrocycle-forming linker connects a pair of amino acids corresponding to amino acids locations 4 and 8 of GHRH 1-29.

15. The method of claim 1 , wherein the second macrocycle-forming linker connects a pair of amino acids corresponding to amino acid locations 21 and 25 of GHRH 1-29.

16. The method of claim 1 , wherein the first macrocycle-forming linker connects a pair of amino acids corresponding to amino acid locations 4 and 8 of GHRH 1-29; and the second macrocycle-forming linker connects a pair of amino acids corresponding to amino acid locations 21 and 25 of GHRH 1-29.

17. The method of claim 1 , wherein each amino acid connected by the at least two macrocycle-forming linkers is an α,α-disubstituted amino acid.

18. The method of claim 1 , wherein the peptidomimetic macrocycle has the formula:

wherein:

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

B is an amino acid or

L is a macrocycle-forming linker of the formula -L 1 -L 2 -;

and wherein A, B, C, D, and E, taken together with the crosslinked amino acids connected by the macrocycle-forming linker L, form the amino acid sequence of the peptidomimetic macrocycle;

R 1 and R 2 are independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;

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

L 1 and L 2 are 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 4 is alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;

each K is 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;

R 7 is —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;

R 8 is —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 and w are independently integers from 1-100;

u is an integer from 1 to 3;

x, y and z are independently integers from 0-10; and

n is an integer from 1-5.

19. The method of claim 18 , wherein L 1 and L 2 are independently alkylene, alkenylene or alkynylene.

20. The method of claim 18 , wherein L 1 and L 2 are independently C 3 -C 10 alkylene or alkenylene.

21. The method of claim 18 , wherein L 1 and L 2 are independently C 3 -C 6 alkylene or alkenylene.

22. The method of claim 18 , wherein R 1 and R 2 are independently H.

23. The method of claim 18 , wherein R 1 and R 2 are independently alkyl.

24. The method of claim 18 , wherein R 1 and R 2 are independently methyl.

25. The method of claim 18 , wherein one of R 1 and R 2 is —H, and the other of R 1 and R 2 is alkyl.

26. The method of claim 18 , wherein the sum of x+y+z is 2, 3, or 6.

27. The method of claim 18 , wherein each of v and w is independently an integer from 1 to 25.

28. The method of claim 18 , wherein A, B, C, D, and E, taken together with the crosslinked amino acids connected by the macrocycle-forming linker L, form an amino acid sequence which is at least about 60% identical to GHRH 1-29.

29. The method of claim 18 , wherein u is 2.

30. The method of claim 29 , wherein the peptidomimetic macrocycle has the formula:

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

B is an amino acid or

L′ is a macrocycle-forming linker of the formula -L 1 ′-L 2 ′-;

and wherein A, B, C, D, and E, taken together with the crosslinked amino acids connected by the macrocycle-forming linkers L and L′, form the amino acid sequence of the peptidomimetic macrocycle;

R 1 ′ and R 2 ′ are independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;

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

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

R 7 ′ is —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;

R 8 ′ is —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′ and w′ are independently integers from 1-100;

x′, y′ and z′ are independently integers from 0-10; and

n is an integer from 1-5.

31. The method of claim 30 , wherein the sum of x′+y′+z′ is 2, 3, or 6.

32. The method of claim 1 , wherein the condition is a muscle wasting disease.

33. The method of claim 1 , wherein the condition is a growth hormone deficiency.

34. The method of claim 1 , wherein the condition is gastroparesis.

35. A method of treating a condition in a subject in need thereof, the method comprising administering to the subject a therapeutically-effective amount of a peptidomimetic macrocycle, wherein the peptidomimetic macrocycle comprises an amino acid sequence of formula:

X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-X24-X25-X26-X27-X28-X29  (SEQ ID NO: 1)

wherein:

X1 is Tyr or His;

X2 is Ala, D-Ala, or Val;

X3 is Asp;

X4 is Ala or a crosslinked amino acid;

X5 is Ile;

X6 is Phe;

X7 is Thr;

X8 is Gln, Asn, or a crosslinked amino acid;

X9 is Ser or a crosslinked amino acid;

X10 is Tyr;

X11 is Arg, Ala or Gln;

X12 is Lys, Ala, Gln or a crosslinked amino acid;

X13 is Val or Ile;

X14 is Leu;

X15 is Gly, Ala or a crosslinked amino acid;

X16 is Gln, Glu or a crosslinked amino acid;

X17 is Leu;

X18 is Ser, Tyr or a crosslinked amino acid;

X19 is Ala or a crosslinked amino acid;

X20 is Arg or Gln;

X21 is Lys, Gln or a crosslinked amino acid;

X22 is Leu, Ala, or a crosslinked amino acid;

X23 is Leu;

X24 is Gln, Glu or His;

X25 is Asp, Glu or a crosslinked amino acid;

X26 is Ile;

X27 is Met, Ile, Leu or Nle;

X28 is Ser or a crosslinked amino acid;

X29 is Arg, Ala, Gln or a crosslinked amino acid;

wherein at least one of the crosslinked amino acids is an α,α-disubstituted amino acid;

wherein the peptidomimetic macrocycle comprises at least one macrocycle-forming linker connecting at least one pair of amino acids selected from X1-X29;

L is a macrocycle-forming linker of the formula -L 1 -L 2 -;

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

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

each K is O, S, SO, SO 2 , CO, or CO 2 ;

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

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

36. The method of claim 35 , wherein the at least one macrocycle-forming linker connects one of the following pairs of amino acids: X4 and X8; X5 and X12; X8 and X12; X8 and X15; X9 and X16; X12 and X16; X12 and X19; X15 and X22; X18 and X25; X21 and X25; X21 and X28; X22 and X29; or X25 and X29.

37. The method of claim 35 , wherein the at least one macrocycle-forming linker connects one of the following pairs of amino acids: X4 and X8; X5 and X12; X12 and X19; X15 and X22; X18 and X25; X21 and X25; or X21 and X28.

38. The method of claim 35 , wherein the condition is a muscle wasting disease.

39. The method of claim 35 , wherein the condition is a growth hormone deficiency.

40. The method of claim 35 , wherein the condition is gastroparesis.

41. The method of claim 35 , wherein the condition is a cachexia.

42. The method of claim 35 , wherein the condition is a lipodystrophy.

43. The method of claim 35 , wherein the condition is a growth hormone disorder.