초록

The present invention provides peptidomimetic macrocycles capable of modulating growth hormone levels and methods of using such macrocycles for the treatment of disease.

대표청구항 ▼

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 -L1-L2-, wherein: each L1 and L2 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene,

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 -L1-L2-, wherein: each L1 and L2 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R4—K—R4—]n, each being optionally substituted with R5;each K is independently O, S, SO, SO2, CO, or CO2;each R4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;each R5 is independently halogen, alkyl, —OR6, —N(R6)2, —SR6, —SOR6, —SO2R6, —CO2R6, a fluorescent moiety, a radioisotope, or a therapeutic agent;each R6 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 -L1-L2-;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 R1 and R2 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;each R3 is independently alkyl, alkenyl, alkynyl, arylalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5;each R7 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5, or part of a cyclic structure with a D residue;each R8 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5, 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; andeach x, y and z is independently an integer from 0-10. 15. The peptidomimetic macrocycle of claim 14, wherein R1 and R2 are independently H. 16. The peptidomimetic macrocycle of claim 14, wherein R1 and R2 are independently alkyl. 17. The peptidomimetic macrocycle of claim 14, wherein R1 and R2 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 -L1′-L2′-;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 R1′ and R2′ is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;each L1′ and L2′ is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R4—K—R4—]n, each being optionally substituted with R5;R7′ is —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5, or part of a cyclic structure with a D residue;R8′ is —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5, or part of a cyclic structure with an E residue;each v′ and w′ is independently an integer from 1-100; andeach 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 -L1-L2-, wherein: each L1 and L2 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R4—K—R4—]n, each being optionally substituted with R5;each K is independently O, S, SO, SO2, CO, or CO2;each R4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;each R5 is independently halogen, alkyl, —OR6, —N(R6)2, —SR6, —SOR6, —SO2R6, —CO2R6, a fluorescent moiety, a radioisotope, or a therapeutic agent;each R6 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heterocycloalkyl, a fluorescent moiety, a radioisotope, or a therapeutic agent; andeach 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 -L1-L2-;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 R1 and R2 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;each R3 is independently alkyl, alkenyl, alkynyl, arylalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5;each R7 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5, or part of a cyclic structure with a D residue;each R8 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5, 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; andeach x, y and z is independently an integer from 0-10. 36. The method of claim 35, wherein R1 and R2 are independently H. 37. The method of claim 35, wherein R1 and R2 are independently alkyl. 38. The method of claim 35, wherein R1 and R2 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 -L1′-L2′-;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 R1′ and R2′ is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-;each L1′ and L2′ is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R4—K—R4—]n, each being optionally substituted with R5;R7′ is —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5, or part of a cyclic structure with a D residue;R8′ is —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, optionally substituted with R5, or part of a cyclic structure with an E residue;each v′ and w′ is independently an integer from 1-100; andeach 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; andii) 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-X29wherein: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 -L1-L2-; each L1 and L2 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R4—K—R4—]n, each being optionally substituted with R5; each R4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene; each K is independently O, S, SO, SO2, CO, or CO2; each R5 is independently halogen, alkyl, —OR6, —N(R6)2, —SR6, —SOR6, —SO2R6, —CO2R6, a fluorescent moiety, a radioisotope, or a therapeutic agent; each R6 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; andii) 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-X29wherein: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 -L1-L2-; each L1 and L2 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R4—K—R4—]n, each being optionally substituted with R5; each R4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene; each K is independently O, S, SO, SO2, CO, or CO2; each R5 is independently halogen, alkyl, —OR6, —N(R6)2, —SR6, —SOR6, —SO2R6, —CO2R6, a fluorescent moiety, a radioisotope, or a therapeutic agent; each R6 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.