Pro-angiogenic peptides and peptide conjugates

Patent metadata

Field	Value
Publication number	US-2016333070-A1
Application number	US-201415105762-A
Country	US
Kind code	A1
Filing date	Dec 17, 2014
Priority date	Dec 20, 2013
Publication date	Nov 17, 2016
Grant date	—

Abstract

Short bioactive sequences derived from the 2 nd loop of the 7-transmembranal receptor of endothelial differentiation gene 3 (EDG3) useful in stimulation of angiogenesis, and peptide conjugates comprising a permeability enhancing moiety, are provided. Also provided are pharmaceutical compositions comprising the peptides and methods of use in conditions were insufficient blood-supply occurs, or which are associated with endothelia dysfunction such as peripheral vascular diseases, coronary artery diseases, cerebrovascular diseases, diabetes and delayed wound healing, pulmonary disease, eye diseases and pathological condition related to severe infection.

First claim

1 .- 50 . (canceled)

51 . A peptide of 3-10 amino acids, comprising the core sequence RPY, wherein R is an arginine residue (Arg) or a modified arginine residue, P is a proline residue (Pro) or a modified proline residue, Y is a tyrosine residue (Tyr) or a modified tyrosine residue, wherein said peptide shares no more than three amino acids that are identical to those of corresponding residues 143-147 of endothelial differentiation gene receptor 3 (EDG3) having the sequence Met-Arg-Pro-Tyr-Asp (SEQ ID NO: 4).

52 . The peptide of claim 51 wherein at least one residue is a non-naturally encoded amino acid residue.

53 . The peptide of claim 52 wherein the non-naturally encoded amino acid residue or modified residue is selected from the group consisting of: Norleucine (Nle), modified Arg, modified Pro and modified Tyr.

54 . The peptide of claim 51 further comprising a permeability-enhancing moiety covalently connected to said peptide via a direct bond or via a linker.

55 . The peptide of claim 54 wherein the linker is a glycine (Gly) residue.

56 . The peptide of claim 54 wherein the permeability-enhancing moiety is a fatty acid residue.

57 . The peptide of claim 51 further comprising the tripeptide Aspartic acid-arginine-tyrosine (Asp-Arg-Tyr) or to an analog thereof.

58 . The peptide of claim 51 wherein the sequence RPY is selected from the group consisting of:
i. Arg-Pro-Tyr;
ii. Arg-Pro-35dITyr;
iii. hArg-Pro-Tyr;
iv. hArg-Pro-35dITyr; and
v. hArg-4Hyp-Tyr

wherein hArg is homoarginine; 35dITyr is 3,5-diiodotyrosine is and 4Hyp is 4hydroxyproline.

59 . A peptide conjugate comprising a peptide according to claim 51 and a moiety capable of increasing permeability, wherein the peptide-conjugate is according to Formula I:
i. Z-X-R-P-Y-B  (Formula I)

wherein Z designates a moiety capable of increasing permeability covalently connected via a direct bond or via a linker; X designates a natural or synthetic amino acid residue other than methionine or X may be absent, R is an arginine residue or a modified R residue, P is a proline residue or a modified proline residue, Y is a tyrosine residue or a modified tyrosine residue, and B designates a terminal carboxy acid, amide, ester or alcohol group.

60 . The peptide conjugate of claim 59 wherein X is a norleucine (Nor) residue, R is selected from the group consisting of an arginine residue, an homoarginine residue (hArg), an N-methyl arginine residue (NMeArg), a citruline residue, and a 2-amino-3-guanidinopropionic acid residue; P is selected from the group consisting of a proline residue, an hydroxyproline residue, and a thiazolidine-carboxylate residue, and Y is selected from the group consisting of: a tyrosine residue, a 3,5 diiodo tyrosine residue (35dITyr), a 3,5 diBromo tyrosine residue (35dBTyr), and an homotyrosine residue.

61 . The peptide conjugate of claim 59 wherein Z designates Myristoyl-glycine (Myr-Gly), X is a norleucine (Nle) residue, R is an arginine residue or an homoarginine residue (hArg), P is a proline residue or 4-hydroxy-Proline residue (4HyP), Y is a tyrosine residue or a (3,5-diiodo)tyrosine (35dITyr) residue and B is terminal designates a terminal carboxy acid, amide, ester or alcohol group.

62 . The peptide conjugate according to claim 59 comprising a peptide selected from the group consisting of:

Gly-Arg-Pro-Tyr-NH2;
(SEQ ID NO: 5)

Gly-Nle-Arg-Pro-Tyr-NH2;
(SEQ ID NO: 6)

Gly-Nle-Arg-Pro-35dITyr-NH2;
(SEQ ID NO: 7)

Gly-Nle-hArg-Pro-Tyr-NH2;
(SEQ ID NO: 8)

Gly-Nle-hArg-Pro-35dITyr-NH2;
(SEQ ID NO: 9)

and

Gly-Nle-hArg-4HyP-Tyr.
(SEQ ID NO: 10)

63 . The peptide conjugate of claim 59 selected from the group consisting of:

Myr-Gly-Arg-Pro-Tyr-NH2;
(SEQ ID NO: 11)

Myr-Gly-Nle-Arg-Pro-Tyr-NH2;
(SEQ ID NO: 12)

Myr-Gly-Nle-Arg-Pro-35dITyr-NH2;
(SEQ ID NO: 13)

Myr-Gly-Nle-hArg-Pro-Tyr-NH2;
(SEQ ID NO: 14)

Myr-Gly-Nle-hArg-Pro-35dITyr-NH2,
(SEQ ID NO: 15)

and

Myr-Gly-Nle-hArg-4HyP-Tyr.
(SEQ ID NO: 16)

64 . A pharmaceutical composition comprising as an active ingredient a peptide according to claim 51 , and a pharmaceutically acceptable carrier.

65 . A pharmaceutical composition comprising as an active ingredient a peptide conjugate according to claim 59 and a pharmaceutically acceptable carrier.

66 . A method of prevention or treatment a condition in which insufficient blood-supply occurs, a condition which is associated with endothelia dysfunction, or a condition mediates through S1P receptor, comprising administering to a subject in need thereof a pharmaceutical composition according to claim 65 .

67 . The method of claim 66 wherein the condition is selected from the group consisting of: peripheral vascular disease, myocardial ischemia, tissue graft, coronary artery diseases, stroke, diabetes, pancreatic islet transplantation, and delayed wound healing, pulmonary disease, eye disease, bone loss and pathological condition related to severe infection.

68 . The method according to claim 67 wherein the disease is selected from the group consisting of: acute lung injury (ALI), acute respiratory distress syndrome (ARDS), and ventilation induced ling injury (VILI); age-related macular disease (AMD); and sepsis.

69 . A stent comprising at least one peptide conjugate according claim 59 .

70 . A method of prevention or treatment of myocardial infection or coronary artery disease, comprising administering to a subject in need thereof a stent according to claim 69 .

Assignees

• Yissum Reseearch Dev Company Of The Hebrew Niversity Of Jerusalem Ltd
• Yissum Res Dev Co

Inventors

• Ben-Sasson Shmuel
• Reuveni Hadas

Classifications

• A61P5/48

  of the pancreatic hormones · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• A61P27/02

  Ophthalmic agents · CPC title