CELL-PENETRATING PEPTIDE SEQUENCES

• US 20190309020A1
• Filed: 11/22/2017
• Published: 10/10/2019
• Est. Priority Date: 11/22/2016
• Status: Active Application

• Current Assignee: Ohio State Innovation Foundation (Ohio State University)
• Original Assignee: Ohio State Innovation Foundation (Ohio State University)
• Inventors: PEI, Dehua, QIAN, Ziqing
• Application Number:
  US16/462,922
• Publication Number:
  US 20190309020A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61K 49/0056 Peptides, proteins, polyamino acids
• CPC Class Codes: C07K 2319/10 containing a tag for extracellular membrane crossing, e.g. TAT or VP22
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/64 Cyclic peptides containing only normal peptide links

Abstract

Disclosed are cell-penetrating cyclic peptides. The peptides can be used to deliver peptides and other biologically active moieties into cells. Formulations containing the cell-penetrating cyclic peptides are also described.

Claims

1. A cyclic peptide according to one of Formula I-A to I-E:

2. The cyclic peptide of claim 1, wherein each hydrophobic amino acid is independently selected from the group consisting of phenylglycine, leucine, isoleucine, noroleucine, methionine, phenylalanine, homophenylalanine, cyclohexylalanine, tyrosine, piperidine-2-carboxylate, tryptophan, proline, 3-(3-benzothienyl)-alanine, and naphthyl alanine, each of which is optionally substituted with one or more substituents.

3. The cyclic peptide of claim 1, wherein each hydrophobic amino acid is independently piperidine-2-carboxylate, naphthylalanine, tryptophan, 3-(3-benzothienyl)-alanine, or phenylalanine, each of which is optionally substituted with one or more substituents.

4. The cyclic peptide of claim 1, wherein at least one hydrophobic amino acid has a hydrophobicity greater than or equal to phenylalanine.

5. The cyclic peptide of claim 1, wherein at least two of the amino acids have the opposite chirality.

6. The cyclic peptide of any of claims 1-5, wherein at least two of the amino acids have the same chirality.

7. The cyclic peptide of any of claims 1-6, wherein one arginine is adjacent to one hydrophobic amino acid.

8. The cyclic peptide of claim 7, wherein the arginine has the same chirality as the adjacent hydrophobic amino acid.

9. The cyclic peptide of any of claims 1-8, wherein at least two arginines are adjacent to each other.

10. The cyclic peptide of any of claims 1-9, wherein three arginines are adjacent to each other.

11. The cyclic peptide of any of claims 1-10, wherein at least two hydrophobic amino acids are adjacent to each other.

12. The cyclic peptide of any of claims 1-11, wherein at least three hydrophobic amino acids are adjacent to each other.

13. The cyclic peptide of any of claims 1-12, wherein any four adjacent amino acids are AA_H2-AA_H1-R-r, AA_H2-AA_H1-r-R, R-r-AA_H1-AA_H2, or r-R-AA_H1-AA_H2, wherein each of AA_H1 and AA_H2 are independently a hydrophobic amino acid.

15. The cyclic peptide of claim 13 or 14, wherein each of AA_H1 and AA_H2 are independently piperidine-2-carboxylate, naphthylalanine, tryptophan, 3-(3-benzothienyl)-alanine, or phenylalanine, each of which is optionally substituted with one or more substituents.

16. The cyclic peptide of any of claims 13-15, wherein each of AA_H1 and AA_H2 are independently a hydrophobic amino acid having a hydrophobicity greater than or equal to phenylalanine.

17. The cyclic peptide of any of claims 13-16, wherein AA_H1 is naphthylalanine.

18. The cyclic peptide of any of claims 13-17, wherein AA_H1 and AA_H2 are naphthylalanine.

19. The cyclic peptide of any of claims 13-18, wherein AA_H1 is a hydrophobic amino acid with a side chain having a large solvent-accessible surface area (SASA).

20. The cyclic peptide of claim 19, wherein the large SASA is at least about 200 Å

².

21. The cyclic peptide of any of claims 13-20, wherein AA_H2 is a hydrophobic amino acid with a side chain having a SASA that is less than or equal to the SASA of the side chain of AA_H1.

22. The cyclic peptide of any of claims 13-21, wherein when any AA_U or any AA_Z is a hydrophobic amino acid, said hydrophobic amino acid has a side chain with SASA which is less than AA_H1.

23. The cyclic peptide of any of claims 13-22, wherein AA_H1 and AA_H2 have the same or opposite chirality.

24. The cyclic peptide of claim 23, wherein AA_H1 and AA_H2 have the opposite chirality.

25. The cyclic peptide of any of claims 13-24, wherein AA_H1 has the same chirality as the adjacent arginine.

26. The cyclic peptide of any of claims 13-25, having one of the following sequences:

AA_H2-AA_H1-R-r-R, AA_H2-AA_H1-R-r-r, AA_H2-AA_H1-r-R-R, AA_H2-AA_H1-r-R-r, R-R-r-AA_H1-AA_H2, r-R-r-AA_H1-AA_H2, r-r-R-AA_H1-AA_H2, or R-r-R-AA_H1-AA_H2.

27. The cyclic peptide of claim 13-26, having one of the following sequences:

AA_H2-LAA_H1-R-r-R, AA_H2-DAA_H1-r-R-r, r-R-r-DAA_H1-AA_H2, or R-r-R-LAA_H1-AA_H2.

28. The cyclic peptide of any of claims 1-27, having a relative cytosolic delivery efficiency which is improved by an amount in the range of from about 50% to about 500% compared to cyclo(FΦ

RRRRQ) (SEQ ID NO;

10).

29. The cyclic peptide of claim 28, wherein the cyclic peptide has a relative cytosolic delivery efficiency which is improved by about 175% to about 250% compared to cyclo(FΦ

RRRRQ) (SEQ ID NO;

10).

30. The cyclic peptide of claim 29, comprising the following sequence:

FfΦ

RrR (SEQ ID NO;

131).

31. The cyclic peptide of claim 28, wherein the cyclic peptide has a relative cytosolic delivery efficiency which is improved by about 150% to about 400% compared to cyclo(FΦ

RRRRQ) (SEQ ID NO;

10).

32. The cyclic peptide of claim 31, comprising the following sequence:

fFϕ

rRr (SEQ ID NO;

132).

33. The cyclic peptide of claim 32, wherein the cyclic peptide has a cytosolic delivery efficiency which is improved by about 75% to about 275% compared to cyclo(FΦ

RRRRQ) (SEQ ID NO;

10).

34. The cyclic peptide of claim 33, comprising the following sequence:

fFfRrR (SEQ ID NO;

133).

35. The cyclic peptide of claim 28, wherein the cyclic peptide has a cytosolic delivery efficiency which is improved by about 150% to about 250% compared to cyclo(FΦ

RRRRQ) (SEQ ID NO;

10).

36. The cyclic peptide of claim 35, comprising the following sequence:

FfFrRr (SEQ ID NO;

134).

37. The cyclic peptide of claim 36, wherein the cyclic peptide has a cytosolic delivery efficiency which is improved by about 200% to about 450% compared to cyclo(FΦ

RRRRQ) (SEQ ID NO;

10).

38. The cyclic peptide of claim 37, comprising the following sequence:

fFϕ

rRr (SEQ ID NO;

135).

39. The cyclic peptide of claim 28, wherein the peptide has a cytosolic delivery efficiency which is improved by about 250% to about 450% compared to cyclo(FΦ

RRRRQ) (SEQ ID NO;

10).

40. The cyclic peptide of claim 39, comprising the following sequence:

fΦ

frRr (SEQ ID NO;

136).

14. A cyclic peptide having a structure according to any of Formula II-A to II-D:

41. A cyclic peptide according to Formula III-A to III-D:

42. The cyclic peptide of claim 41, wherein each hydrophobic amino acid is independently selected from the group consisting of phenylglycine, leucine, isoleucine, noroleucine, methionine, phenylalanine, homophenylalanine, cyclohexylalanine, tyrosine, piperidine-2-carboxylate, tryptophan, proline, 3-(3-benzothienyl)-alanine, and naphthyl alanine, each of which is optionally substituted with one or more substituents.

43. The cyclic peptide of claim 41, wherein each hydrophobic amino acid is independently piperidine-2-carboxylate, naphthylalanine, tryptophan, 3-(3-benzothienyl)-alanine, or phenylalanine, each of which is optionally substituted with one or more substituents.

44. The cyclic peptide of claim 41, wherein at least one hydrophobic amino acid has a hydrophobicity greater than or equal to phenylalanine.

45. The cyclic peptide of claim 41, wherein at least two of the amino acids have the opposite chirality.

46. The cyclic peptide of any of claims 41-45, wherein at least two of the amino acids have the same chirality.

47. The cyclic peptide of any of claims 41-46, wherein one arginine is adjacent to one hydrophobic amino acid.

48. The cyclic peptide of claim 47, wherein the arginine has the same chirality as the adjacent hydrophobic amino acid.

49. The cyclic peptide of any of claims 41-48, wherein at least two arginines are adjacent to each other.

50. The cyclic peptide of any of claims 41-49, wherein three arginines are adjacent to each other.

51. The cyclic peptide of any of claims 41-50, wherein at least two hydrophobic amino acid are adjacent to each other.

52. The cyclic peptide of any of claims 41-51, wherein at least three hydrophobic amino acid are adjacent to each other.

53. The cyclic peptide of any of claims 41-52, wherein any four adjacent amino acids are AA_H2-AA_H1-R-r, AA_H2-AA_H1-r-R, R-r-AA_H1-AA_H2, or r-R-AA_H1-AA_H2, wherein each of AA_H1 and AA_H2 are independently a hydrophobic amino acid.

54. The cyclic peptide of any of claims 41-52, wherein the peptide as a structure according to any of Formula IV-A to IV-P:

55. The cyclic peptide of claim 52 or 54, wherein each of AA_H1 and AA_H2 are independently piperidine-2-carboxylate, naphthylalanine, tryptophan, 3-(3-benzothienyl)-alanine, or phenylalanine, each of which is optionally substituted with one or more substituents.

56. The cyclic peptide of any of claims 53-55, wherein each of AA_H1 and AA_H2 are independently a hydrophobic amino acid having a hydrophobicity greater than or equal to phenylalanine.

57. The cyclic peptide of any of claims 53-56, wherein AA_H1 is naphthylalanine.

58. The cyclic peptide of any of claims 53-57, wherein AA_H and AA_H2 are naphthylalanine.

59. The cyclic peptide of any of claims 53-57, wherein AA_H1 is a hydrophobic amino acid with a side chain having a large solvent-accessible surface area (SASA).

60. The cyclic peptide of claim 59, wherein the large SASA is at least about 200 Å

².

61. The cyclic peptide of any of claims 53-50, wherein when any AA_U or any AA_Z is a hydrophobic amino acid, said hydrophobic amino acid has a side chain with SASA which is less than AA_H1.

62. The cyclic peptide of any of claims 53-61, wherein the cyclic peptide does not include a hydrophobic amino acid with a side chain having SASA which is greater than AA_H1.

63. The cyclic peptide of any of claims 53-62, wherein AA_H1 and AA_H2 have the same or opposite chirality.

64. The cyclic peptide of claim 63, wherein AA_H1 and AA_H2 have the opposite chirality.

65. The cyclic peptide of any of claims 53-64, wherein AA_H1 has the same chirality as the adjacent arginine.

66. The cyclic peptide of any of claims 53-65, having one of the following sequences:

AA_H2-AA_H1-R-r-R, AA_H2-AA_H1-R-r-r, AA_H2-AA_H1-r-R-R, AA_H2-AA_H1-r-R-r, R-R-r-AA_H1-AA_H2, r-R-r-AA_H1-AA_H2, r-r-R-AA_H1-AA_H2, or, R-r-R-AA_H1-AA_H2.

67. The cyclic peptide of claim 53-66, having one of the following sequences AA_H2-LAA_H1-R-r-R, AA_H2-DAA_H1-r-R-r, r-R-r-DAA_H1-AA_H2, or R-r-R-LAA_H1-AA_H2.

86. A method of treating or diagnosing a patient in need thereof, comprising administering cyclic peptide according to any of claims 41-67.

87. A composition comprising the cyclic peptide according to any of claims 41-67.

68. A peptide comprising Formula V-A to V-D:

69. The peptide of claim 68, wherein an amino group on the N-terminus or a carboxylate group on the C-terminus of any of Formula V-A to V-D independently form a peptide bond.

70. The peptide of claim 68, further comprising a cyclization moiety, wherein the cyclization moiety, which thereby forms a cyclic peptide.

71. The peptide of claim 70, wherein the cyclization moiety independently forms a peptide bond with an amino group on the N-terminus or a carboxylate group on the C-terminus of any of Formula V-A to V-D.

72. The peptide of any of claims 68-71, further comprising a cargo moiety (X_n), wherein at least one atom or bond is replaced by a bond to X_n.

73. The cyclic peptide of any of claims 68-72, wherein each of AA_H1 and AA_H2 are independently piperidine-2-carboxylate, naphthyl alanine, tryptophan, 3-(3-benzothienyl)-alanine, or phenylalanine, each of which is optionally substituted with one or more substituents.

74. The cyclic peptide of any of claims 68-73, wherein each of AA_H1 and AA_H2 are independently a hydrophobic amino acid having a hydrophobicity greater than or equal to phenylalanine.

75. The cyclic peptide of any of claims 68-74, wherein AA_H1 is naphthylalanine.

76. The cyclic peptide of any of claims 68-75, wherein AA_H1 and AA_H2 are naphthylalanine.

77. The cyclic peptide of any of claims 68-76, wherein AA_H1 is a hydrophobic amino acid with a side chain having a large solvent-accessible surface area (SASA).

78. The cyclic peptide of claim 77, wherein the large SASA is at least about 200 Å

².

79. The cyclic peptide of any of claims 68-78, wherein AA_H2 is a hydrophobic amino acid with a side chain having a SASA that is less than or equal to the SASA of the hydrophobic side chain of AA_H1.

80. The cyclic peptide of any of claims 68-79, wherein when any AA_U or any AA_Z is a hydrophobic amino acid, said hydrophobic amino acid has a side chain with SASA which is less than AA_H1.

81. The cyclic peptide of any of claims 68-80, wherein AA_H1 and AA_H2 have the same or opposite chirality.

82. The cyclic peptide of claim 81, wherein AA_H1 and AA_H2 have the opposite chirality.

83. The cyclic peptide of any of claims 68-82, wherein AA_H1 has the same chirality as the adjacent arginine.

84. The cyclic peptide of any of claims 68-83, having one of the following sequences:

AA_H2-AA_H1-R-r-R, AA_H2-AA_H1-R-r-r, AA_H2-AA_H1-r-R-R, AA_H2-AA_H1-r-R-r, R-R-r-AA_H1-AA_H2, r-R-r-AA_H1-AA_H2, r-r-R-AA_H1-AA_H2, or, R-r-R-AA_H1-AA_H2.

85. The cyclic peptide of claim 68-84, having one of the following sequences AA_H2-LAA_H1-R-r-R, AA_H2-DAA_H1-r-R-r, r-R-r-DAA_H1-AA_H2, or R-r-R-LAA_H1-AA_H2.

88. The peptide of any of claims 68-85, wherein the cytosolic delivery efficiency of the peptide comprising any of Formula V-A to V-D is improved by an amount in the range of from about 50% to about 500% compared to cyclo(FΦ

RRRRQ) (SEQ ID NO;

10).

89. The peptide of claim 88, wherein the peptide comprising any of Formula V-A to V-D comprises the following sequence:

FfΦ

RrR (SEQ ID NO;

131).

90. The peptide of claim 88, wherein the peptide comprising any of Formula V-A to V-D comprises the following sequence:

fFϕ

rRr (SEQ ID NO;

132).

91. The peptide of claim 88, wherein the peptide comprising any of Formula V-A to V-D comprises the following sequence:

fFfRrR (SEQ ID NO;

133).

92. The peptide of claim 88, wherein the peptide comprising any of Formula V-A to V-D comprises the following sequence:

FfFrRr (SEQ ID NO;

134).

93. The peptide of claim 88, wherein the peptide comprising any of Formula V-A to V-D comprises the following sequence:

fFϕ

rRr (SEQ ID NO;

135).

94. The peptide of claim 88, wherein the peptide comprising any of Formula V-A to V-D comprises the following sequence:

fΦ

frRr (SEQ ID NO;

136).