Peptidomimetic macrocycles
The present invention provides peptidomimetic macrocycles capable of modulating growth hormone levels and methods of using such macrocycles for the treatment of disease.
1. A peptidomimetic macrocycle comprising an amino acid sequence which is at least about 60% identical to GHRH 1-29, and a crosslinker that is a macrocycle-forming linker of the formula -L 1 -L 2 -, wherein:
each L 1 and L 2 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 K is independently O, S, SO, SO 2 , CO, or CO 2 ;
each R 4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;
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; and each n is independently an integer from 1-5, wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid.
2. The peptidomimetic macrocycle of claim 1 , wherein the macrocycle-forming linker connects a first amino acid and a second amino acid, wherein the first amino acid and the second amino acid are selected from amino acids corresponding to 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; and 25 and 29 of GHRH 1-29.
3. The peptidomimetic macrocycle of claim 1 , wherein the macrocycle-forming linker connects a first amino acid and a second amino acid, wherein the first amino acid and the second amino acid are selected from amino acids corresponding to the following locations of amino acids: 4 and 8; 5 and 12; 12 and 19; 15 and 22; 18 and 25; 21 and 25; and 21 and 28 of GHRH 1-29.
4. The peptidomimetic macrocycle of claim 1 , wherein the macrocycle-forming linker connects a first amino acid and a second amino acid, wherein the first amino acid and the second amino acid correspond to the following locations of amino acids: 4 and 8 of GHRH 1-29.
5. The peptidomimetic macrocycle of claim 1 , wherein the macrocycle-forming linker connects a first amino acid and a second amino acid, wherein the first amino acid and the second amino acid correspond to the following locations of amino acids: 21 and 25 of GHRH 1-29.
6. The peptidomimetic macrocycle of claim 1 , comprising two macrocycle-forming linkers.
7. The peptidomimetic macrocycle of claim 1 , comprising an amino acid sequence which is at least about 60% identical to an amino acid sequence chosen from the group consisting of the amino acid sequences in Tables 1, 2 or 4.
8. The peptidomimetic macrocycle 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 the amino acid sequences in Tables 1, 2 or 4.
9. The peptidomimetic macrocycle 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 the amino acid sequences in Tables 1, 2 or 4.
10. The peptidomimetic macrocycle of claim 1 , wherein the peptidomimetic macrocycle comprises a helix.
11. The peptidomimetic macrocycle of claim 1 , wherein the peptidomimetic macrocycle comprises an α-helix.
12. The peptidomimetic macrocycle of claim 1 , wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid.
13. The peptidomimetic macrocycle of claim 1 , wherein the macrocycle forming linker connects two α,α-disubstituted amino acids.
14. The peptidomimetic macrocycle of claim 1 , wherein the peptidomimetic macrocycle has the formula:
wherein:
each A, C, D, and E is independently an amino acid;
each B is independently an amino acid or
each L is independently a macrocycle-forming linker of the formula -L 1 -L 2 -;
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;
each R 1 and R 2 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;
each R 3 is independently alkyl, alkenyl, alkynyl, arylalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R 5 ;
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;
each v and w is independently an integer from 1-100;
u is an integer from 1 to 3; and
each x, y and z is independently an integer from 0-10.
15. The peptidomimetic macrocycle of claim 14 , wherein R 1 and R 2 are independently H.
16. The peptidomimetic macrocycle of claim 14 , wherein R 1 and R 2 are independently alkyl.
17. The peptidomimetic macrocycle of claim 14 , wherein R 1 and R 2 are independently methyl.
18. The peptidomimetic macrocycle of claim 14 , wherein the sum of x+y+z is 2, 3, or 6.
19. The pharmaceutical composition of claim 14 , wherein the peptidomimetic macrocycle has the formula:
L′ is a macrocycle-forming linker of the formula -L 1 ′-L 2 ′-;
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;
each R 1 ′ and R 2 ′ is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;
each L 1 ′ and L 2 ′ is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R 4 —K—R 4 —] n , each being optionally substituted with R 5 ;
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;
each v′ and w′ is independently an integer from 1-100; and
each x′, y′ and z′ is independently an integer from 0-10.
20. The peptidomimetic macrocycle of claim 19 , wherein the sum of x′+y′+z′ is 2, 3 or 6.
21. A pharmaceutical composition comprising the peptidomimetic macrocycle of claim 1 .
22. 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, and a crosslinker that is a macrocycle-forming linker of the formula -L 1 -L 2 -, wherein:
each L 1 and L 2 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 K is independently O, S, SO, SO 2 , CO, or CO 2 ;
each R 4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;
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; and
each n is independently an integer from 1-5, wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid.
23. The method of claim 22 , wherein the macrocycle-forming linker connects a first amino acid and a second amino acid, wherein the first amino acid and the second amino acid are selected from amino acids corresponding to 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; and 25 and 29 of GHRH 1-29.
24. The method of claim 22 , wherein the macrocycle-forming linker connects a first amino acid and a second amino acid, wherein the first amino acid and the second amino acid are selected from amino acids corresponding to the following locations of amino acids: 4 and 8; 5 and 12; 12 and 19; 15 and 22; 18 and 25; 21 and 25; and 21 and 28 of GHRH 1-29.
25. The method of claim 22 , wherein the macrocycle-forming linker connects a first amino acid and a second amino acid, wherein the first amino acid and the second amino acid correspond to the following locations of amino acids: 4 and 8 of GHRH 1-29.
26. The method of claim 22 , wherein the macrocycle-forming linker connects a first amino acid and a second amino acid, wherein the first amino acid and the second amino acid correspond to the following locations of amino acids: 21 and 25 of GHRH 1-29.
27. The method of claim 22 , wherein the peptidomimetic macrocycle comprises two macrocycle-forming linkers.
28. The method of claim 22 , wherein the peptidomimetic macrocycle comprises an amino acid sequence which is at least about 60% identical to an amino acid sequence chosen from the group consisting of the amino acid sequences in Tables 1, 2, or 4.
29. The method of claim 22 , 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 the amino acid sequences in Tables 1, 2, or 4.
30. The method of claim 22 , 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 the amino acid sequences in Tables 1, 2, or 4.
31. The method of claim 22 , wherein the peptidomimetic macrocycle comprises a helix.
32. The method of claim 22 , wherein the peptidomimetic macrocycle comprises an α-helix.
33. The method of claim 22 , wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid.
34. The method of claim 22 , wherein the macrocycle forming linker connects two α,α-disubstituted amino acids.
35. The method of claim 22 , wherein the peptidomimetic macrocycle has the formula:
wherein:
each A, C, D, and E is independently an amino acid;
each B is independently an amino acid or
each L is independently a macrocycle-forming linker of the formula -L 1 -L 2 -;
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;
each R 1 and R 2 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;
each R 3 is independently alkyl, alkenyl, alkynyl, arylalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R 5 ;
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;
each v and w is independently an integer from 1-100;
u is an integer from 1 to 3; and
each x, y and z is independently an integer from 0-10.
36. The method of claim 35 , wherein R 1 and R 2 are independently H.
37. The method of claim 35 , wherein R 1 and R 2 are independently alkyl.
38. The method of claim 35 , wherein R 1 and R 2 are independently methyl.
39. The method of claim 35 , wherein the sum of x+y+z is 2, 3, or 6.
40. The method of claim 35 , wherein the peptidomimetic macrocycle has the formula:
L′ is a macrocycle-forming linker of the formula -L 1 ′-L 2 ′-;
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;
each R 1 ′ and R 2 ′ is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;
each L 1 ′ and L 2 ′ is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R 4 —K—R 4 —] n , each being optionally substituted with R 5 ;
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;
each v′ and w′ is independently an integer from 1-100; and
each x′, y′ and z′ is independently an integer from 0-10.
41. The method of claim 40 , wherein the sum of x′+y′+z′ is 2, 3, or 6.
42. The method of claim 22 , wherein the condition is a cachexia.
43. The method of claim 42 , wherein the cachexia is cancer cachexia, chronic heart failure cachexia, chronic obstructive pulmonary disease cachexia, or rheumatoid arthritis cachexia.
44. The method of claim 22 , wherein the condition is a lipodystrophy.
45. The method of claim 44 , wherein the lipodystrophy is a HIV lipodystrophy.
46. The method of claim 22 , wherein the condition is a growth hormone disorder.
47. The method of claim 46 , wherein the growth hormone disorder is an adult growth hormone deficiency.
48. The method of claim 46 , wherein the growth hormone disorder is a pediatric growth hormone deficiency.
49. The method of claim 48 , wherein the pediatric growth hormone deficiency is associated with idiopathic short stature, SGA (infant small for gestational age), chronic kidney disease, Prader-Willi syndrome Turner syndrome, short stature homeobox (SHOX) gene deficiency, or primary IGF-1 deficiency.
50. The method of claim 22 , wherein the condition is gastroparesis/short bowel syndrome.
51. 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, comprising at least two macrocycle-forming linkers, wherein the first of said two macrocycle-forming linkers connects a first amino acid to a second amino acid, and the second of said 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.
52. 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:
i) an amino acid sequence which is at least about 60% identical to GHRH 1-29; and
ii) a macrocycle-forming linker connecting a first amino acid to a second amino acid, wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid, and wherein the first and second amino acids are selected from amino acids corresponding to 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; and 22 and 29 of GHRH 1-29.
53. 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
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 the peptidomimetic macrocycle comprises at least one macrocycle-forming linker L connecting at least one pair of amino acids;
each L is independently a macrocycle-forming linker of the formula -L 1 -L 2 -;
each L 1 and L 2 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 4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;
each K is independently 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 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; and
each n is independently an integer from 1-5, wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid.
54. A pharmaceutical composition comprising a peptidomimetic macrocycle, wherein the peptidomimetic macrocycle comprises an amino acid sequence which is at least about 60% identical to GHRH 1-29, comprising at least two macrocycle-forming linkers, wherein the first of said two macrocycle-forming linkers connects a first amino acid to a second amino acid, and the second of said 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.
55. A pharmaceutical composition comprising a peptidomimetic macrocycle, wherein the peptidomimetic macrocycle comprises:
i) an amino acid sequence which is at least about 60% identical to GHRH 1-29; and
ii) a macrocycle-forming linker connecting a first amino acid to a second amino acid, wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid, and wherein the first and second amino acids are selected from amino acids corresponding to 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; and 22 and 29 of GHRH 1-29.
56. A pharmaceutical composition comprising 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
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 the peptidomimetic macrocycle comprises at least one macrocycle-forming linker L connecting at least one pair of amino acids;
each L is independently a macrocycle-forming linker of the formula -L 1 -L 2 -;
each L 1 and L 2 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 4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;
each K is independently 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 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; and
each n is independently an integer from 1-5, wherein the peptidomimetic macrocycle comprises an α,α-disubstituted amino acid.