阅读说明：本技术 用于治疗糖尿病的肽 (Peptides for the treatment of diabetes ) 是由 J·阿伦法尔 P·杜纳 A·豪塔特尼尔森 B·瓦尔斯 于 2018-05-04 设计创作，主要内容包括：本发明涉及剂及其在治疗哺乳动物的内分泌、营养和/或代谢疾病中的用途。本发明还涉及新型肽。(The present invention relates to agents and their use in the treatment of endocrine, nutritional and/or metabolic disorders in mammals. The invention also relates to novel peptides.)

1. An agent, comprising:

a) A peptide or peptide analog, wherein the peptide or peptide analog comprises an amino acid sequence of the general formula:

KXLAXXXXIXLXYGIK(SEQ ID NO:140)

Wherein:

x2 is C, P or G;

X5 is E or G;

x6 is C, D or I;

X7 is D, I, S or G;

X8 is S, D or G;

X10 is E or G;

x12 is S or T;

provided that if X12 is T, then the peptide comprises no more than 25 amino acid residues; and

Provided that if X2 is P, X5 is E, X6 is I, X7 is D, X8 is S, X10 is E and X12 is S, then the peptide comprises no more than 85 amino acid residues;

Or a biologically active fragment and/or variant of said peptide or peptide analogue selected from the group consisting of CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEGDIELSYGIK (SEQ ID NO:147), KPLAEIELSYGIK (SEQ ID NO:148), KCLAEIDSCELSYGIK (SEQ ID NO:155) and CFKPLAEIDSIEC (SEQ ID NO: 156);

b) a polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c).

2. The agent of claim 1, wherein the peptide or peptide analog comprises an amino acid sequence of the general formula:

KXLAXXXXIXLSYGIK(SEQ ID NO:162)

wherein:

X2 is C, P or G;

X5 is E or G;

X6 is C, I or absent;

x7 is D, G or absent;

X8 is S, G or absent;

X10 is E or G.

3. The agent according to any one of the preceding claims, wherein the agent comprises no more than 85, such as no more than 80, such as no more than 75, such as no more than 70, such as no more than 65, such as no more than 60, such as no more than 55, such as no more than 50, such as no more than 55, such as no more than 40 amino acids, such as no more than 35, such as no more than 30, such as no more than 28, such as no more than 26, such as no more than 24, such as no more than 22, such as no more than 20, such as no more than 19, such as no more than 18, such as no more than 17, such as no more than 16, such as no more than 15, such as no more than 14, such as no more than 13, such as no more than 12, such as no more than 11, such as no more than.

4. An agent according to any one of the preceding claims, wherein the agent comprises at least 2 additional amino acids, such as at least 3, such as at least 4, such as at least 5, such as at least 6, such as at least 7, such as at least 8, such as at least 9, such as at least 10, such as at least 15, or such as at least 20 amino acids, conjugated to the N-or C-terminus of the peptide.

5. An agent according to any preceding claim, wherein the agent is non-naturally occurring.

6. an agent according to any preceding claim wherein the agent is conjugated to a moiety.

7. The agent of claim 6, wherein the moiety is selected from the group consisting of polyethylene glycol (PEG), monosaccharides, fluorophores, chromophores, radioactive compounds, and cell penetrating peptides.

8. An agent according to any preceding claim, wherein the agent is further modified, for example glycosylated, or modified by pegylation, amidation, esterification, acylation, acetylation and/or alkylation.

9. an agent according to any preceding claim, wherein the agent comprises or consists of a tandem repeat sequence.

10. The agent according to claim 9, wherein the tandem repeat sequence comprises or consists of the amino acid sequence of any one or more of the sequences recited in the preceding claims.

11. an agent according to any preceding claim wherein the agent is fused to another polypeptide.

12. The agent of claim 11, wherein the polypeptide is selected from the group consisting of glutathione-S-transferase (GST) and protein a.

13. the agent according to any one of the preceding claims, wherein the agent is fused to a tag.

14. the agent of claim 13, wherein the tag is an oligo-histidine tag.

15. the agent according to any one of the preceding claims, wherein the agent is cyclic.

16. An agent as claimed in any preceding claim wherein the peptide or peptide analogue is capable of forming at least one intramolecular cysteine bridge.

17. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises an amino acid sequence selected from the group consisting of: KCLAECDSIELSYGIK (SEQ ID NO:141), CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEDISIELSYGIK (SEQ ID NO:145), KPLAEIGDIELSYGIK (SEQ ID NO:146), KPLAEGDIELSYGIK (SEQ ID NO:147), KPLAEIELSYGIK (SEQ ID NO:148), KPLAEIDSIELTYGIK (SEQ ID NO:149), KPLAEIDGIELSYGIK (SEQ ID NO:150), KPLAEIDGIELTYGIK (SEQ ID NO:151), KPLAEIGSIELSYGIK (SEQ ID NO:152), KGLAEIDSIELSYGIK (SEQ ID NO:153), KPLAGIDSIGLSYGIK (SEQ ID NO:154), KCLAEIDSCELSYGIK (SEQ ID NO:155) and CFKPLAEIDSIEC (SEQ ID NO:156), or a variant or fragment thereof.

18. An agent according to any preceding claim, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAEIDSIELSYGIK (SEQ ID NO:136), or a variant or fragment thereof.

19. An agent according to any preceding claim, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAGIDSIGLSYGIK (SEQ ID NO:154), or a variant or fragment thereof.

20. An agent according to any preceding claim, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KGLAEIDSIELSYGIK (SEQ ID NO:153), or a variant or fragment thereof.

21. An agent according to any preceding claim, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KCLAECDSIELSYGIK (SEQ ID NO:141), or a variant or fragment thereof.

22. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAEIDGIELTYGIK (SEQ ID NO:151), or a variant or fragment thereof.

23. An agent according to any preceding claim, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAEIGSIELSYGIK (SEQ ID NO:152), or a variant or fragment thereof.

24. An agent according to any preceding claim, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAEIELSYGIK (SEQ ID NO:148), or a variant or fragment thereof.

25. An agent according to any one of the preceding claims wherein the variant comprises or consists of a sequence in which any one amino acid has been changed to another proteinogenic amino acid or to a non-proteinogenic amino acid, provided that no more than 5 amino acids have been so changed.

26. an agent according to any one of the preceding claims, wherein the variant comprises or consists of a sequence in which no more than 5 amino acids, such as no more than 4 amino acids, such as no more than 3 amino acids, such as no more than 2 amino acids, such as no more than 1 amino acid, are changed to another proteinogenic amino acid or a non-proteinogenic amino acid.

27. an agent according to any preceding claim wherein one or more amino acids are conservatively substituted.

28. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of one or more additional amino acids inserted at the N-and/or C-terminus and/or within the sequence.

29. an agent as claimed in any preceding claim wherein the peptide or peptide analogue has 1 additional amino acid.

30. The agent of any one of the preceding claims, wherein the agent further comprises a detectable moiety.

31. The agent of any one of the preceding claims, wherein the detectable moiety comprises or consists of a radioisotope.

32. The agent according to any one of the preceding claims, wherein the radioisotope is selected from the group consisting of 99mTc, 111In, 67Ga, 68Ga, 72As, 89Zr, 123I and 201 Tl.

33. An agent according to any preceding claim wherein the detectable moiety is detectable by imaging techniques such as SPECT, PET, MRI, optical imaging or ultrasound imaging.

34. Use of an agent according to any one of the preceding claims in the manufacture of a diagnostic composition for diagnosing a disease, disorder or injury of the pancreas in an individual.

35. A composition comprising the agent of any one of the preceding claims.

36. the composition of claim 35, wherein the composition is a pharmaceutical composition.

37. An agent, comprising:

c) A peptide or peptide analog comprising or consisting of the amino acid sequence GDPNDGRGDSVVYGLR (SEQ ID NO:137), VDTYDGGISVVYGLR (SEQ ID NO:138), VDTYDGDGSVVYGLR (SEQ ID NO:139), VDVPEGDISLAYGLR (SEQ ID NO:157), LDGLVRAYDNISPVG (SEQ ID NO:158), GDPNGDISVVYGLR (SEQ ID NO:159), VDVPNGDISLAYRLR (SEQ ID NO:160), VDVPEGDISLAYRLR (SEQ ID NO:161), V (. beta. -D) TYDGDISVVYGLR (SEQ ID NO:167), VDTY (. beta. -D) GDISVVYGLR (SEQ ID NO:168), VDTYDG (. beta. -D) ISVYGLR (SEQ ID NO: 169);

b) a polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c).

38. An agent or composition according to any preceding claim for use as a medicament.

39. An agent, comprising:

a) A peptide or peptide analog selected from the group consisting of:

(i) A peptide comprising or consisting of an amino acid sequence of the general formula:

KXLAXXXXIXLXYGIK(SEQ ID NO:140)

Wherein the content of the first and second substances,

x2 is C, P or G;

X5 is E or G;

X6 is C, D or I;

X7 is D, I, S or G;

x8 is S, D or G;

X10 is E or G;

x12 is S or T;

provided that if X12 is T, then the peptide comprises no more than 25 amino acid residues;

(ii) a peptide comprising, or consisting of, an amino acid sequence of the formula:

VDZZZGZZSZZYGLR(SEQ ID NO:68)

Wherein the content of the first and second substances,

Z3 is T or V;

Z4 is Y or P;

Z5 is D or N;

Z7 is D or G;

z8 is I or G;

z10 is V or L;

z11is V or A; and

(iii) a peptide comprising an amino acid sequence selected from the group consisting of: KCLAECDSIELSYGIK (SEQ ID NO:141), CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEGDIELSYGIK (SEQ ID NO:147), KPLAEIELSYGIK (SEQ ID NO:148), KCLAEIDSCELSYGIK (SEQ ID NO:155) and CFKPLAEIDSIEC (SEQ ID NO:156), or consist thereof;

b) a polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c);

for use in the treatment of endocrine, nutritional and/or metabolic disorders in a mammal.

40. Agent or composition for use according to any one of the preceding claims, wherein said agent comprises a second or additional active ingredient.

41. The agent or composition for use according to claim 40, wherein the second or additional active ingredient is selected from the group consisting of insulin, glucagon-like peptide-1 (GLP-1), sulfonylureas, dipeptidyl peptidase-4 (DPP4) inhibitors, alpha-glucosidase inhibitors, thiazolidinediones, meglitinide and sodium-glucose cotransporter-2 (SGLT2) inhibitors.

42. An agent or composition according to any preceding claim for use in the treatment of endocrine, nutritional and/or metabolic disorders in a mammal.

43. The agent or composition for use according to claim 42, wherein the mammal is a human.

44. Agent or composition for use according to any of the preceding claims, wherein the endocrine, nutritional and/or metabolic disease is selected from the group consisting of diabetes, type 1 diabetes, type 2 diabetes, diabetes related to malnutrition, glucose regulation and pancreatic endocrine disorders, insulin resistance syndrome, impaired glucose tolerance, hyperglycemia, hyperinsulinemia and any combination thereof.

45. Agent or composition for use according to any of the preceding claims, wherein the endocrine, nutritional and/or metabolic disease is selected from the group consisting of diabetes, thyroid disorders, glucose regulation and pancreatic endocrine disorders, endocrine gland disorders, nutritional deficiencies, obesity, overnutrition and metabolic disorders.

46. Agent or composition for use according to any of the preceding claims, wherein the diabetes is selected from the group consisting of type 1 diabetes, type 2 diabetes, diabetes related to nutritional disorders, diabetes with explicit indication and diabetes without explicit indication.

47. Agent or composition for use according to any one of the preceding claims, wherein the glucose regulation and pancreatic endocrine disorders are selected from the group consisting of non-diabetic hypoglycaemic coma and pancreatic endocrine disorders.

48. agent or composition for use according to any of the preceding claims, wherein the obesity and overnutrition disorder is selected from the group consisting of local obesity, overnutrition, and overnutrition sequelae.

49. Agent or composition for use according to any of the preceding claims, wherein the condition of nutritional deficiency is selected from the group consisting of disorders of aromatic amino acid metabolism, disorders of branched chain amino acid metabolism and fatty acid metabolism, disorders of amino acid metabolism, lactose intolerance, disorders of carbohydrate metabolism, disorders of sphingolipid metabolism, disorders of lipid storage, disorders of glycosaminoglycan metabolism, disorders of glycoprotein metabolism, disorders of lipoprotein metabolism, lipemia, disorders of purine and pyrimidine metabolism, disorders of porphyrin and bilirubin metabolism, disorders of mineral metabolism, cystic fibrosis, amyloidosis, hypovolemia, disorders of body fluids, electrolytes and acid-base balance, and post-operative endocrine and metabolic disorders.

50. A method of treating endocrine, nutritional, and/or metabolic diseases, the method comprising administering to a subject in need thereof an agent according to any one of the preceding claims.

51. use of an agent according to any one of the preceding claims in the manufacture of a medicament for the treatment of endocrine, nutritional and/or metabolic disorders in a mammal.

52. A method for delaying the onset of diabetes and/or a diabetes-related condition or disease comprising administering to an individual in need thereof a therapeutically effective amount of the agent of any one of the preceding claims.

53. A method for lowering blood glucose levels, comprising administering to an individual in need thereof a therapeutically effective amount of the agent of any one of the preceding claims.

54. the method of claim 53, wherein insulin secretion is increased.

55. The method of claim 53, wherein uptake of glucose by the cell is increased.

56. the method of claim 53, wherein insulin production is increased.

57. The method of claim 53, wherein glucagon production is reduced.

58. A method of improving beta cell viability, comprising administering to a subject in need thereof a therapeutically effective amount of the agent of any one of the preceding claims.

59. A method for improving the morphology of beta cells, comprising administering to an individual in need thereof a therapeutically effective amount of the agent of any one of the preceding claims.

60. a method of stabilizing or improving islet viability and/or morphology comprising administering to an individual in need thereof a therapeutically effective amount of the agent of any one of the preceding claims.

Technical Field

The present invention relates to peptides for use in the treatment of diabetes and related disorders.

Background

the peptide hormone insulin, produced by the beta cells in the islets of langerhans in the pancreas, is released in response to an increase in blood glucose levels. Thus, glucose is removed from the blood by insulin-dependent stimulation of glucose transporters located in the cell membranes of target tissues (e.g., adipose tissue, skeletal muscle, and liver). Insulin exerts its biological effects by binding to and activating membrane-bound Insulin Receptors (IR), thereby initiating a cascade of intracellular signaling events to regulate a variety of biological processes such as glycolipid metabolism.

Currently, the treatment of diabetes (both type 1 and type 2 diabetes) relies primarily on insulin therapy. Supplements to insulin therapy are long-acting glucagon-like peptide-1 (GLP-1) receptor agonists, i.e., derivatives that act at the same receptor as GLP-1. GLP-1 is a metabolic hormone that promotes insulin secretion. In addition to increasing insulin secretion by the pancreas in a glucose-dependent manner, GLP-1 is known to increase insulin sensitivity in both alpha-cells and beta cells; increase beta cell mass and insulin expression, post-translational modifications and secretion; and decrease glucagon secretion by the pancreas. Other drugs used to assist insulin therapy to lower blood glucose levels include DPP-IV inhibitors, metformin, SGLT-2 inhibitors and sulfonylureas.

Certain disadvantages such as weight gain and increased risk of cancer and hypoglycemia are associated with long-term insulin use. Accordingly, there is an increasing need in the art for new non-insulin compounds that are not only capable of treating diabetes by addressing insulin resistance and hyperglycemia, but also reduce the associated and subsequent complications.

Therefore, it would be of great interest to identify new compounds that are capable of restoring glucose metabolism and treating diabetes and related disorders. A variety of methods are contemplated, although none of these methods will be apparent to those skilled in the art.

Disclosure of Invention

The present inventors have discovered peptides that stimulate the proliferation of beta cells, have the ability to rescue beta cells from glycotoxic condition-induced apoptosis, and stimulate insulin secretion from rat INS-1 beta cells and ex vivo mouse islets. Furthermore, the present inventors found that, in a glucose tolerance test, the peptide lowers the blood glucose level in vivo and delays the onset of diabetic disease in BB lyp/lyp rats (a model of type 1 diabetes). Thus, the peptides of the invention are useful in the treatment of endocrine, nutritional and metabolic diseases and disorders.

In one aspect, the invention relates to an agent comprising or consisting of:

a) A peptide or peptide analog, wherein the peptide or peptide analog comprises an amino acid sequence of the general formula:

KXLAXXXXIXLXYGIK(SEQ ID NO:140)

Wherein the content of the first and second substances,

X2 is C, P or G;

X5 is E or G;

x6 is C, D or I;

X7 is D, I, S or G;

X8 is S, D or G;

x10 is E or G;

x12 is S or T;

Provided that if X12 is T, then the peptide comprises no more than 25 amino acids; and

provided that if X2 is P, X5 is E, X6 is I, X7 is D, X8 is S, X10 is E and X12 is S, then the peptide comprises no more than 85 amino acid residues;

Or a biologically active fragment and/or variant of said peptide or peptide analog, wherein said biologically active fragment and/or variant is selected from the group consisting of: CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEGDIELSYGIK (SEQ ID NO:147), KPLAEIELSYGIK (SEQ ID NO:148), KCLAEIDSCELSYGIK (SEQ ID NO:155), and CFKPLAEIDSIEC (SEQ ID NO: 156);

b) A polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) a cell comprising the polynucleotide of b) or the vector of c).

In one aspect, the present invention relates to an agent comprising:

a) A peptide or peptide analogue comprising or consisting of the amino acid sequence: GDPNDGRGDSVVYGLR (SEQ ID NO:137), VDTYDGGISVVYGLR (SEQ ID NO:138), and VDTYDGDGSVVYGLR (SEQ ID NO:139), VDVPEGDISLAYGLR (SEQ ID NO:157), LDGLVRAYDNISPVG (SEQ ID NO:158), GDPNGDISVVYGLR (SEQ ID NO:159), VDVPNGDISLAYRLR (SEQ ID NO:160) VDVPEGDISLAYRLR (SEQ ID NO:161), V (. beta. -D) TYDGDISVVYGLR (SEQ ID NO:167), VDTY (. beta. -D) GDISVVYGLR (SEQ ID NO:168), VDTYGDG (. beta. -D) ISVYGLR (SEQ ID NO: 169);

b) A polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c).

In one aspect, the present invention relates to compositions comprising the above agents.

In one aspect, the invention relates to an agent or a composition comprising said agent for use as a medicament.

In one aspect, the present invention relates to an agent comprising:

a) (ii) (i) a peptide or peptide analogue, wherein the peptide or peptide analogue comprises or consists of an amino acid sequence of the general formula:

KXLAXXXXIXLXYGIK(SEQ ID NO:140)

wherein the content of the first and second substances,

X2 is C, P or G;

X5 is E or G;

X6 is C, D or I;

X7 is D, I, S or G;

X8 is S, D or G;

X10 is E or G;

X12 is S or T;

Provided that if X12 is T, then the peptide comprises no more than 25 amino acid residues;

(ii) A peptide, wherein the peptide comprises an amino acid sequence of the general formula:

VDZZZGZZSZZYGLR(SEQ ID NO:68)

Wherein the content of the first and second substances,

Z3 is T or V;

z4 is Y or P;

z5 is D or N;

Z7 is D or G;

Z8 is I or G;

Z10 is V or L;

Z11is V or A;

(iii) a peptide, wherein the peptide comprises or consists of an amino acid sequence selected from the group consisting of KCLAECDSIELSYGIK (SEQ ID NO:141), CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEIELSYGIK (SEQ ID NO:148), KCLAEIDSCELSYGIK (SEQ ID NO:155), and CFKPLAEIDSIEC (SEQ ID NO: 156);

b) A polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c);

the agents are useful for treating endocrine, nutritional and/or metabolic disorders in a mammal.

In one aspect, the invention relates to a method for treating an endocrine, nutritional, and/or metabolic disease comprising administering to an individual in need thereof a therapeutically effective amount of an agent described herein.

In one aspect, the invention relates to the use of an agent as described herein for the preparation of a medicament for the treatment of endocrine, nutritional and/or metabolic disorders.

In one aspect, the invention relates to a method of delaying the onset of diabetes comprising administering to an individual in need thereof a therapeutically effective amount of an agent described herein.

In one aspect, the invention relates to a method of reducing blood glucose levels comprising administering to an individual in need thereof a therapeutically effective amount of an agent described herein.

In one aspect, the invention relates to a method, e.g., an in vitro method, for improving beta cell morphology comprising administering to an individual in need thereof a therapeutically effective amount of an agent described herein.

In one aspect, the invention relates to a method for improving beta cell viability comprising administering to an individual in need thereof a therapeutically effective amount of an agent described herein.

In one aspect, the invention relates to the use of an agent as described herein for the preparation of a diagnostic composition for diagnosing a disease, disorder or injury of the pancreas in an individual.

drawings

FIG. 1 FOL-005 and FOL-014 induced proliferation of beta cells

The addition of FOL-005 in increasing concentrations to the solution induced an increase in the proliferation of INS-1 cells after 48 hours (FIG. 1A). Wells coated with FOL-005 and blocked with Bovine Serum Albumin (BSA) induced more proliferation of β -cells compared to control (ctrl) wells coated with BSA only (fig. 1B). Wells pre-coated with FOL-014 and blocked with BSA induced more proliferation than wells coated with BSA alone (fig. 1C). Data are expressed as Counts Per Minute (CPM) relative to unstimulated control (ctrl) cells. The mean + -SD of 10-12 different observations per group is given.

FIG. 2 FOL-005 protected beta cells against glycotoxicity

INS-1 cells cultured for 48 hours in 20mM glucose showed more apoptotic cells (annexin V positive) than cells cultured in 5mM glucose. Addition of FOL-005 to cells cultured with 20mM glucose reduced the level of apoptotic cells compared to 20mM glucose alone (FIG. 2A). Increased apoptosis as measured by Caspase-3 activity at 20mM in INS-1 cells compared to 5mM glucose. Addition of FOL-005 decreased the rate of glycotoxicity-induced Caspase-3 activity (FIG. 2B). The mean ± SD of 4-8 different observations per group is given.

FIG. 3 increase of insulin secretion from pancreatic islets and beta cells following FOL-005 stimulation

FOL-005 stimulates insulin secretion by beta cells and pancreatic islets. INS-1 cells released increased insulin following stimulation with FOL-005 (6. mu.M) in glucose-free medium compared to unstimulated control (ctrl) and interfering control peptide (FOL-015) (FIG. 3A). FOL-005 stimulated INS-1 release insulin at low (5mM) and high (20mM) glucose (FIG. 3B). Isolated mouse islets stimulated with FOL-005 (6. mu.M) or GLP-1(100nM) secreted more insulin than unstimulated control islets (FIG. 3C). The mean ± SD of 5-6 different observations per group is given.

FIG. 4 increase of insulin secretion from pancreatic islets and beta cells following FOL-014 stimulation

FOL-014 stimulated insulin secretion by beta cells and pancreatic islets. INS-1 cells stimulated with FOL-014 (6. mu.M) secreted more insulin than unstimulated control cells (FIG. 4A). Ex vivo mouse islets stimulated with FOL-014(6 μ M) secreted more insulin than control islets (fig. 4B). The addition of GLP-1(100nM) or FOL-014 (0.6. mu.M) had no effect on insulin secretion. The mean ± SD of 5-6 different observations per group is given.

FIG. 5 the effect of FOL-014 on insulin secretion was dose-dependent. Stimulation of INS-1 cells by increasing doses of FOL-014 resulted in a significant increase in insulin secretion at all concentrations tested. Insulin secretion increased in a linear fashion in the presence of FOL-014 ranging from 0.6nM to 60 nM. Higher concentrations appear to result in less pronounced effects on insulin secretion. Furthermore, FOL-014 induced insulin secretion comparable to that of 100nM GLP-1. Bars represent mean and Standard Error of Mean (SEM).

FIG. 6 the effect of FOL-014 on insulin secretion is glucose concentration dependent. Insulin secretion by INS-1 cells untreated or exposed to FOL-014 was measured in the presence of increased glucose concentration. At glucose levels of 5.5mM or higher, insulin secretion was significantly higher in FOL-014 treated cells compared to untreated control cells. Bars represent mean and Standard Error of Mean (SEM).

FIG. 7 FOL-005 and FOL-014 resulted in additive effects on insulin secretion in combination with native GLP-1. Insulin release from INS-1 cells was measured after treatment of GLP-1 in combination with FOL-005 and FOL-014 (all three peptides at a concentration of 100nM), respectively, and compared to the effect of each peptide alone. The combination of GLP-1 and FOL-014 significantly increased insulin secretion compared to each peptide alone. An increase was also observed for the combination of FOL-005 and GLP-1. Bars represent mean and Standard Error of Mean (SEM).

FIG. 8. Effect of FOL-014 on insulin and glucagon secretion in pancreatic islets. FOL-014 at two different concentrations was tested for its effect on ex vivo mouse islets in glucose at low (2.8mM) (A, C) and high (16.7mM) (B, D) concentrations and compared to 100nM GLP-1. In the low glucose samples, the presence of FOL-014 did not increase insulin secretion but decreased glucagon secretion compared to control and GLP-1. In the high glucose samples, 600nM FOL-014 and GLP-1, but not 6 μ M FOL-014, significantly increased insulin secretion (B), and GLP-1 and FOL-014 at both concentrations were effective in decreasing glucagon secretion (d). Bars represent mean and Standard Error of Mean (SEM).

FIG. 9 FOL-014 reduced blood glucose levels in vivo after glucose injection. The intraperitoneal glucose tolerance test (IPGTT) was performed on wild-type C57bl/6 mice. FOL-014 administered at 200nmol/kg significantly reduced blood glucose levels at 15, 30 and 45 minutes (P ═ 0.0027) compared to controls. At a dose of 30nmol/kg, FOL-014 reduced glucose levels at 45 minutes post-glucose injection and had a significant effect. The dashed line corresponds to the mean non-fasting glucose level. Data represent mean and Standard Error of Mean (SEM). Statistical analysis was performed using student's t-test.

FIG. 10 FOL-014 delaying the onset of type 1 diabetes in BB lyp/lyp rats. BB lyp/lyp rats treated with FOL-014 showed a significant delay in the onset of diabetes, defined as plasma glucose <11.1 mmol/l. The age of onset of diabetes for each rat is depicted in (a), where there is a significant difference between the untreated and treated groups. The percentage of animals with daily progression to type 1 diabetes is depicted in (B) with significant differences between groups. (A) Error bars in (1) indicate standard error of the mean (SEM).

FIG. 11. Effect of peptide analogs derived from FOL-005 or FOL-014 on insulin secretion. The ability of the novel peptide analogues to induce insulin secretion under high glucose (16.7mM) conditions was tested in two separate INS-1 cell lines (a and B). This effect was compared with the effects of native GLP-1, FOL-005 and FOL-014, and the effect of high glucose alone. Analogs that induced insulin release below the average of the high glucose controls were considered non-functional (not shown). Insulin secretion levels of the novel analogs are indicated by black filled bars and used for comparison in the comparative pattern. Bars represent mean and Standard Error of Mean (SEM).

FIG. 12 shows specific distribution patterns after mice were injected with FOL-005 and FOL-014. Following subcutaneous administration of 3H-FOL-005, the highest overall radioactivity levels were present at the pancreas and injection site 1 hour (a) and 2 hours (B) post injection. Accumulation of 3H-FOL-005 was also observed in liver, kidney, and salivary glands. In vivo biodistribution and tissue localization in NMRI nude mice of Cy7.5-labeled FOL-005(C) and FOL-014(D) via subcutaneous injection were studied using the Pearl Trilogy small animal imaging system. After the initial control imaging, each mouse was administered a dose of 10nmol and real-time imaging was performed at 5 minutes, 20 minutes, 50 minutes, 60 minutes, 2 hours, 4 hours, 6 hours, 24 hours, and 48 hours. High accumulation of both peptides was evident in the pancreatic region and at the injection site.

Detailed Description

The invention is as defined in the claims.

In one aspect, the invention relates to a peptide or peptide analog comprising an amino acid sequence of the general formula:

a)KXLAXXXXIXLXYGIK(SEQ ID NO:140)

wherein:

X2 is C, P or G;

X5 is E or G;

x6 is C, D or I;

x7 is D, I, S or G;

X8 is S, D or G;

X10 is E or G;

X12 is S or T;

Provided that if X12 is T, then the peptide comprises no more than 25 amino acids; and

provided that if X2 is P, X5 is E, X6 is I, X7 is D, X8 is S, X10 is E and X12 is S, then the peptide comprises no more than 85 amino acid residues;

b) A polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c).

In one embodiment, the invention relates to a peptide or peptide analog comprising an amino acid sequence of the general formula:

KXLAXXXXIXLSYGIK(SEQ ID NO:162)

Wherein:

x2 is C, P or G;

X5 is E or G;

x6 is C, I or absent;

x7 is D, G or absent;

X8 is S, G or absent;

X10 is E or G;

Wherein absent is amino acid X5 linked to amino acid X10.

In one embodiment, the invention relates to a peptide comprising an amino acid sequence of the general formula:

KXLAXIXLSYGIK(SEQ ID NO:163)

Wherein:

X2 is C, P or G;

X5 is E or G;

X10 is E or G.

In one embodiment, the present invention relates to an agent comprising:

a) A peptide, wherein the peptide is selected from the group consisting of:

i) A peptide comprising or consisting of the amino acid sequence: 136, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156;

ii) a biologically active sequence variant of any one of the peptides of i), wherein any one amino acid has been changed to another proteinogenic amino acid or to a non-proteinogenic amino acid, with the proviso that no more than 5 amino acids have been so changed;

iii) a biologically active fragment of any one of the peptides of i) or ii), wherein said fragment comprises at least 10 contiguous amino acids of any one of i) or ii);

b) A polynucleotide encoding the peptide of a) when expressed;

c) a vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c).

In one embodiment, the present invention relates to an agent comprising:

a) A peptide, wherein the peptide comprises or consists of an amino acid sequence selected from the group consisting of GDPNDGRGDSVVYGLR (SEQ ID NO:137), VDTYDGGISVVYGLR (SEQ ID NO:138), and VDTYDGDGSVVYGLR (SEQ ID NO:139), VDVPEGDISLAYGLR (SEQ ID NO:157), LDGLVRAYDNISPVG (SEQ ID NO:158), GDPNGDISVVYGLR (SEQ ID NO:159), VDVPNGDISLAYRLR (SEQ ID NO:160), VDVPEGDISLAYRLR (SEQ ID NO: 161);

b) a polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c).

in one embodiment, the present invention relates to a peptide comprising an amino acid sequence of the general formula:

VDVPZGDISLAYZLR(SEQ ID NO:164)，

Wherein:

z5 is E or N;

Z13 is R or G.

In one embodiment, the present invention relates to a peptide comprising an amino acid sequence of the general formula:

VDTYDGZZSVVYGLR(SEQ ID NO:165)

wherein:

Z7 is D or G;

Z8 is I or G.

In one embodiment, the present invention relates to a peptide comprising an amino acid sequence of the general formula:

GDPNZZZZZSVVYGLR(SEQ ID NO:166)

Wherein:

Z5 is D or G;

Z6 is D or G;

z7 is I or R;

Z8 is G or absent;

Z9 is D or absent.

The term "absent" as used herein, e.g., "X6 is C, I or absent" is understood to mean that the amino acid residues directly adjacent to the absent amino acid are directly linked to each other by a conventional amide bond.

The term "peptide analog" as used herein refers to a peptide comprising or consisting of a non-naturally occurring peptide.

The term "amino acid" as used herein includes the standard 20 genetically encoded amino acids and their corresponding stereoisomers of the 'D' form (as compared to the natural 'L' form), omega-and other naturally occurring amino acids, unconventional amino acids (e.g., alpha-disubstituted amino acids, N-alkyl amino acids, etc.) and chemically derivatized amino acids (see below).

when specifically recited amino acids (e.g., 'alanine' or 'Ala' or 'a'), the term refers to both L-alanine and D-alanine, unless specifically stated otherwise. Other non-conventional amino acids may also be suitable components of the peptides of the invention, as long as the peptide retains the desired functional properties. For the peptides shown, each encoded amino acid residue is represented by a single letter designation, corresponding to the common name of a conventional amino acid, where appropriate.

Chemical derivatives of one or more amino acids may be achieved by reaction with a functional side group. These derivatives include, for example, those molecules in which a reactive (free) amino group is derivatized to form the hydrochloride salt of an amine, p-toluenesulfonyl, carboxyphenylmethoxy, tert-butoxycarbonyl, chloroacetyl, or formyl. The reactable carboxyl groups can be derivatized to form salts, methyl and ethyl esters, or other types of esters and hydrazides. The reactable hydroxyl group may be derivatized to form an O-acyl or O-alkyl derivative. Also included as chemical derivatives are those peptides that are naturally occurring amino acid derivatives containing twenty standard amino acids. For example: 4-hydroxyproline can replace proline; 5-hydroxy lysine can be substituted for lysine; 3-methylhistidine can replace histidine; homoserine can replace serine and ornithine can replace lysine. Derivatives also include peptides containing one or more additions or deletions, so long as the requisite activity is maintained. Other included modifications are terminal modifications such as amidation, amino terminal acylation (e.g. acetylation or thioglycolic acid amidation), terminal carboxyamidation (e.g. with ammonia or methylamine), and the like.

Some peptides of the invention share amino acid sequence similarity with a subdomain of naturally occurring osteopontin. In some embodiments, the peptide may be considered an active fragment or variant, e.g., a variant of a fragment, of a naturally occurring osteopontin.

Some peptides of the invention share amino acid sequence similarity with a naturally occurring subregion of tenascin. In some embodiments, the peptide may be considered an active fragment or variant of a naturally occurring tenascin, e.g., a variant of a fragment.

A "fragment" comprises at least 5 contiguous amino acids of an amino acid sequence, for example at least 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 contiguous amino acids of an amino acid sequence. Thus, the fragment may be 15 or fewer amino acids in length, for example 14, 13, 12,11, 10, 9, 8, 7, 6, or 5 amino acids in length.

In one embodiment, the peptide is no more than 85, such as no more than 80, such as no more than 75, such as no more than 70, such as no more than 65, such as no more than 60, such as no more than 55, such as no more than 50, such as no more than 55, such as no more than 40 amino acids, such as no more than 35, such as no more than 30, such as no more than 28, such as no more than 26, such as no more than 24, such as no more than 22, such as no more than 20, such as no more than 19, such as no more than 18, such as no more than 17, such as no more than 16, such as no more than 15, such as no more than 14, such as no more than 13, such as no more than 12, such as no more than 11, such as no more than 10 amino acids in length.

in another embodiment, the peptide is 5 to 30, such as 5 to 20, such as 8 to 16, such as 10 to 15 amino acids in length.

In another embodiment, the fragment comprises amino acids that are 15 or less amino acids in length, such as less than 14 amino acids, such as less than 13 amino acids, such as less than 12 amino acids, such as less than 11 amino acids, such as less than 10 amino acids, such as less than 9 amino acids, such as less than 8 amino acids, such as less than 7 amino acids, such as less than 6 amino acids, such as less than 5 amino acids.

The term "variant" refers to a peptide that does not have 100% amino acid sequence identity to the parent peptide, i.e., one or more amino acids must be mutated. "mutation" refers to an alteration of an amino acid at a specified position in the parent peptide. For example, the amino acid at a given position may be deleted, changed, substituted or may be a site where one or more amino acids are inserted/added. One skilled in the art will appreciate that substitutions may be conservative or non-conservative.

In one embodiment, the peptide variant comprises or consists of a sequence in which no more than 5 amino acids (e.g., no more than 4 amino acids, e.g., no more than 3 amino acids, e.g., no more than 2 amino acids, e.g., no more than 1 amino acid) are changed to another proteinogenic amino acid or non-proteinogenic amino acid. In one embodiment, one or more amino acids may be conservatively substituted. "conservative substitution" refers to the replacement of one amino acid with another having similar properties (size, hydrophobicity, etc.) such that the function of the peptide is not significantly altered. Thus, "conservative substitutions" refer to contemplated combinations, e.g., Gly, Ala; val, Ile, Leu; asp and Glu; asn, Gln; ser, Thr; lys, Arg; and Phe, Tyr.

In another embodiment, the peptide comprises or consists of one or more additional amino acids inserted at the N-terminus and/or C-terminus and/or interior of the sequence. In one embodiment, at least 2 additional amino acids are inserted, such as at least 3, such as at least 4, such as at least 5, such as at least 6, such as at least 7, such as at least 8, such as at least 9, such as at least 10, such as at least 15, or such as at least 20 additional amino acids. The additional amino acid may be an amino acid from the corresponding position of wild-type human osteopontin (SEQ ID NO: 66) or an amino acid from the corresponding position of wild-type murine osteopontin (SEQ ID NO: 134). The term "corresponding position" of wild-type osteopontin means that the additional amino acids are identical to the amino acids present at equivalent positions in the wild-type osteopontin described above (if one imagines the amino acid sequence of SEQ ID NO:1 replacing the sequence underlined in italics in SEQ ID NO: 66).

In another embodiment, the peptide is selected from the group consisting of: 1, 136, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 67, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 119, 110, 111, 112, 111, 112, 125, 122, 116, 122, 116, 124, 122, 116, 126, 121, 122, 124, 122, 124, 129. 130, 131, 132, 133, 135, 137, 138, 139, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 167, 168, and 169;

i.15-amino acid peptide:

ii.14-amino acid peptide:

iii.13-amino acid peptide:

12-amino acid peptide:

v.11-amino acid peptide:

10-amino acid peptide:

vii.9-amino acid peptides:

viii.8-amino acid peptide:

ix.7-amino acid peptide:

x.6-amino acid peptide:

xi.5-amino acid peptide:

xii.16-amino acid peptide:

xiii.15-amino acid peptide:

xiv.14-amino acid peptide:

xv.13-amino acid peptide:

xvi.12-amino acid peptide:

xvii.11-amino acid peptide:

xviii.10-amino acid peptide:

xix.9-amino acid peptide:

xx.8-amino acid peptides:

xxi.7-amino acid peptide:

xxii.6-amino acid peptide:

xxiii.5-amino acid peptides:

xxiv.16-amino acid peptide:

xxv.15-amino acid peptide:

xxvi.16-amino acid peptide:

xxvii.8-amino acid peptides:

xxviii.18-amino acid peptide:

xxix.16-amino acid peptide:

xxx.15-amino acid peptide:

xxxi.13-amino acid peptide:

xxxii.16-amino acid peptide:

xxxiii.13-amino acid peptide:

xxxiv.15-amino acid peptide:

xxxv.14-amino acid peptide:

xxxvi.15-amino acid peptide:

in one embodiment, the peptide is derived from osteopontin, e.g., a mammalian osteopontin variant and/or fragment.

In one embodiment, the peptide is non-naturally occurring, e.g., a peptide comprising non-protein amino acid residues.

In some embodiments, the peptide is further conjugated to a moiety (moiey) that may be selected from the group consisting of PEG, monosaccharides, fluorophores, chromophores, radioactive compounds, and cell penetrating peptides. In one embodiment, the fluorophore is selected from the group consisting of: luciferin, biotin, 5, 6-carboxytetramethylrhodamine (TAMRA), indocyanine dicarbocyanine (C5), Alexa488, Alexa532, Alexa647, ATTO 488, ATTO 532, 6-carboxyfluorescein (6-FAM), Alexa350, DY-415, ATTO 425, ATTO 465, FL, isothiocyanatofluorescein, Oregon Green 488 (Oregon488), Oregon Green 514 (Oregon514), Rhodamine Green TM (Rhodamine5' -tetrachlorofluorescein, ATTO 520, 6-carboxy-4 ',5' -dichloro-2 ',7' -dimethoxyfluorescein, subunit Malachium TM (Yakima Yellow TM) dye, 530/550, hexachlorofluorescein, Alexa555, DY-549, TMR-X, cyanine phosphoramidite (cyanine 3, cyanine 3.5, cyanine 5.5, cyanine 7.5), Rhodamine T565), Rhodamine Red (ATTO 565), carboxy-X-rhodamine, Texas Red (sulforhodamine 101 acid chloride), Red 610, ATTO 594, DY-480-XL, DY-610, ATTO 610, Red 640, Bodipy 630/650, ATTO 633, Bodipy 650/665, ATTO 647N, DY-649, Red 670, ATTO 680, Red 705, DY-682, ATTO 700, ATTO 740, DY-782, IRD 700, IRD 800, CALCId 540nm, CALCId 522nm, CALCId 544nm, CAOran560 nm, CALorange 538nm, CALorange 559nm, CALRed 590nm, CALRed 9nm, CALRed 591nm, CALRed 610nm, CALRed 590nm, CALRed 610nm, CALRed 635nm, 570nm, 566nm, DY 3, 548nm, Dy, 690nm, 705, Dyd670 nm, Cy 5.

In another embodiment, the peptide is further modified, for example by glycosylation, or by pegylation, amidation, esterification, acylation, acetylation, and/or alkylation.

In one embodiment, the peptide comprises, or consists of, a tandem repeat sequence, which may comprise, or consist of, an amino acid sequence of any one or more of the sequences described herein.

In one embodiment, the peptide is cyclic. The cyclic structure may be achieved by any suitable synthetic method. Thus, heterocyclic linkages may include, but are not limited to, formation through disulfide, cysteine, alkylene, or sulfide bridges.

In a further embodiment, the peptide comprises or consists of a fusion. For example, the peptide may comprise SEQ ID NO:1 or 136.

The term "fusion" of a peptide relates to a polypeptide corresponding to, for example, SEQ ID NO:1 or 136 (or a fragment or variant thereof). For example, the peptide may be fused to a polypeptide such as glutathione-S-transferase (GST) or protein A to facilitate purification of the peptide. Examples of such fusions are well known to those skilled in the art. Similarly, the peptide may be fused to an oligo-histidine tag (e.g. His6), or to an epitope recognized by an antibody (e.g. the well-known Myc tag epitope). Fusions to any variant or derivative of the peptide are also included within the scope of the invention.

alternatively, the fusion moiety may be a lipophilic molecule or peptide domain capable of promoting cellular uptake of the polypeptide, as known to those skilled in the art.

Novel peptides

In one embodiment, the present invention relates to a peptide comprising or consisting of an amino acid sequence selected from the group consisting of: KPLAEIDSIELSYGIK (SEQ ID NO:136), GDPNDGRGDSVVYGLR (SEQ ID NO:137), VDTYDGGISVVYGLR (SEQ ID NO:138), and VDTYDGDGSVVYGLR (SEQ ID NO:139), VDVPEGDISLAYGLR (SEQ ID NO:157), LDGLVRAYDNISPVG (SEQ ID NO:158), GDPNGDISVVYGLR (SEQ ID NO:159), VDVPNGDISLAYRLR (SEQ ID NO:160), VDVPEGDISLAYRLR (SEQ ID NO: 161).

In another embodiment, the invention relates to a peptide comprising or consisting of an amino acid sequence selected from the group consisting of: KCLAECDSIELSYGIK (SEQ ID NO:141), CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEDISIELSYGIK (SEQ ID NO:145), KPLAEIGDIELSYGIK (SEQ ID NO:146), KPLAEGDIELSYGIK (SEQ ID NO:147), KPLAEIELSYGIK (SEQ ID NO:148), KPLAEIDSIELTYGIK (SEQ ID NO:149), KPLAEIDGIELSYGIK (SEQ ID NO:150), KPLAEIDGIELTYGIK (SEQ ID NO:151), KPLAEIGSIELSYGIK (SEQ ID NO:152), KGLAEIDSIELSYGIK (SEQ ID NO:153), KPLAGIDSIGLSYGIK (SEQ ID NO:154), KCLAEIDSCELSYGIK (SEQ ID NO:155) and CFKPLAEIDSIEC (SEQ ID NO: 156).

In one embodiment, the invention relates to an agent comprising a peptide, wherein said peptide comprises or consists of amino acid sequence KPLAEIDSIELSYGIK (SEQ ID NO:136), or a variant or fragment thereof.

In one embodiment, the invention relates to an agent comprising a peptide, wherein the peptide comprises or consists of amino acid sequence KPLAGIDSIGLSYGIK (SEQ ID NO:154), or a variant or fragment thereof.

In one embodiment, the invention relates to an agent comprising a peptide, wherein said peptide comprises or consists of amino acid sequence KGLAEIDSIELSYGIK (SEQ ID NO:153), or a variant or fragment thereof.

In one embodiment, the invention relates to an agent comprising a peptide, wherein said peptide comprises or consists of amino acid sequence KCLAECDSIELSYGIK (SEQ ID NO:141), or a variant or fragment thereof.

In one embodiment, the invention relates to an agent comprising a peptide, wherein said peptide comprises or consists of amino acid sequence KPLAEIDGIELTYGIK (SEQ ID NO:151), or a variant or fragment thereof.

In one embodiment, the invention relates to an agent comprising a peptide, wherein the peptide comprises or consists of amino acid sequence KPLAEIGSIELSYGIK (SEQ ID NO:152), or a variant or fragment thereof.

In one embodiment, the invention relates to an agent comprising a peptide, wherein the peptide comprises or consists of amino acid sequence KPLAEIELSYGIK (SEQ ID NO:148), or a variant or fragment thereof.

In one embodiment, the present invention relates to an agent comprising:

a) a peptide or peptide analogue comprising or consisting of the amino acid sequence: GDPNDGRGDSVVYGLR (SEQ ID NO:137), VDTYDGGISVVYGLR (SEQ ID NO:138), and VDTYDGDGSVVYGLR (SEQ ID NO:139), VDVPEGDISLAYGLR (SEQ ID NO:157), LDGLVRAYDNISPVG (SEQ ID NO:158), GDPNGDISVVYGLR (SEQ ID NO:159), VDVPNGDISLAYRLR (SEQ ID NO:160), VDVPEGDISLAYRLR (SEQ ID NO:161), V (. beta. -D) TYDGDISVVYGLR (SEQ ID NO:167), VDTY (. beta. -D) GDISVVYGLR (SEQ ID NO:168), VDTYGDG (. beta. -D) ISVYGLR (SEQ ID NO: 169);

b) A polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) a cell comprising the polynucleotide of b) or the vector of c).

In some embodiments, the variant comprises or consists of a sequence in which any one amino acid has been changed to another proteinogenic amino acid or non-proteinogenic amino acid, provided that such change is no more than 5 amino acids, such as no more than 4 amino acids, such as no more than 3 amino acids, such as no more than 2 amino acids, such as no more than 1 amino acid. In some embodiments, one or more amino acids are conservatively substituted.

in some embodiments, the peptide comprises or consists of one or more additional amino acids inserted at the N-terminus and/or C-terminus and/or interior of the sequence. In one embodiment, at least 2 additional amino acids may be inserted, such as at least 3, such as at least 4, such as at least 5, such as at least 6, such as at least 7, such as at least 8, such as at least 9, such as at least 10, such as at least 15, or such as at least 20 additional amino acids.

in some embodiments, the peptide is no more than 85, such as no more than 80, such as no more than 75, such as no more than 70, such as no more than 65, such as no more than 60, such as no more than 55, such as no more than 50, such as no more than 55, such as no more than 40 amino acids, such as no more than 35, such as no more than 30, such as no more than 28, such as no more than 26, such as no more than 24, such as no more than 22, such as no more than 20, such as no more than 19, such as no more than 18, such as no more than 17, such as no more than 16, such as no more than 15, such as no more than 14, such as no more than 13, such as no more than 12, such as no more than 11, such as no more than 10 amino acids in.

in some embodiments, the peptide is further conjugated to a moiety that may be selected from the group consisting of PEG, monosaccharides, fluorophores, chromophores, radioactive compounds, and cell penetrating peptides.

In one embodiment, the peptide is further modified, for example by glycosylation, or by pegylation, amidation, esterification, acylation, acetylation, and/or alkylation.

In some embodiments, the peptide comprises or consists of a tandem repeat, wherein the tandem repeat may comprise or consist of an amino acid sequence of any one or more of the sequences described above.

In one embodiment, the peptide is cyclic. The cyclic structure may be achieved by any suitable synthetic method. Thus, heterocyclic linkages may include, but are not limited to, formation through disulfide, cysteine, alkylene, or sulfide bridges.

Indications of

The agents of the invention are useful in the treatment of endocrine, nutritional and metabolic diseases and disorders.

In one embodiment, the mammal in need of treatment for endocrine, nutritional and/or metabolic disorders is a human.

in some embodiments, the endocrine, nutritional, and/or metabolic disease is selected from the group consisting of diabetes, type 1 diabetes, type 2 diabetes, diabetes related to malnutrition, glucose regulation and pancreatic endocrine disorders, insulin resistance syndrome, impaired glucose tolerance, hyperglycemia, hyperinsulinemia, and any combination thereof.

in some embodiments, the endocrine, nutritional, and/or metabolic disease is selected from the group consisting of diabetes, thyroid disorders, glucose regulation, and pancreatic endocrine disorders, endocrine gland disorders, malnutrition, nutrient deficiency, obesity, nutrient excess, and metabolic disorders.

In one embodiment, the diabetes is selected from the group consisting of type 1 diabetes, type 2 diabetes, diabetes related to malnutrition, manifest (specified) diabetes, and non-manifest (unspecified) diabetes.

In one embodiment, the glucose regulation and pancreatic endocrine disorder is selected from the group consisting of non-diabetic hypoglycemic coma and pancreatic endocrine disorder.

In one embodiment, the condition of obesity and overnutrition is selected from the group consisting of localized obesity, overnutrition and the sequelae of overnutrition.

in one embodiment, the condition of nutritional deficiency is selected from the group consisting of disorders of aromatic amino acid metabolism, disorders of branched chain amino acid metabolism and fatty acid metabolism, disorders of amino acid metabolism, lactose intolerance, disorders of carbohydrate metabolism, disorders of sphingolipid metabolism, disorders of lipid storage, disorders of glycosaminoglycan metabolism, disorders of glycoprotein metabolism, disorders of lipoprotein metabolism, lipidemia (lipiemia), disorders of purine and pyrimidine metabolism, disorders of porphyrin and bilirubin metabolism, disorders of mineral metabolism, cystic fibrosis, amyloidosis, hypovolemia, disorders of body fluid, electrolyte and acid-base balance, and post-operative endocrine and metabolic disorders.

Composition comprising a metal oxide and a metal oxide

In one aspect, the present invention relates to a composition comprising an agent as described herein.

In one aspect, the invention relates to an agent selected from the group consisting of:

a) A peptide or peptide analog selected from the group consisting of:

a peptide comprising or consisting of an amino acid sequence of the general formula:

KXLAXXXXIXLXYGIK(SEQ ID NO:140)

wherein the content of the first and second substances,

X2 is C, P or G;

X5 is E or G;

X6 is C, D or I;

x7 is D, I, S or G;

X8 is S, D or G;

X10 is E or G;

X12 is S or T;

Provided that if X12 is T, then the peptide comprises no more than 25 amino acids; and

(ii) a peptide comprising or consisting of an amino acid sequence of the general formula:

VDZZZGZZSZZYGLR(SEQ ID NO:68)

Wherein the content of the first and second substances,

Z3 is T or V;

Z4 is Y or P;

Z5 is D or N;

Z7 is D or G;

Z8 is I or G;

Z10 is V or L;

Z11is V or A; and

(iii) a peptide comprising an amino acid sequence selected from the group consisting of: KCLAECDSIELSYGIK (SEQ ID NO:141), CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEIELSYGIK (SEQ ID NO:148), KCLAEIDSCELSYGIK (SEQ ID NO:155) and CFKPLAEIDSIEC (SEQ ID NO:156), or consist thereof;

b) A polynucleotide encoding the peptide of a) when expressed;

c) a vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c);

the agents are useful for treating endocrine, nutritional and/or metabolic disorders in a mammal.

in one aspect, the invention relates to a composition for treating endocrine, nutritional, and/or metabolic diseases comprising an agent described herein. In one embodiment, the composition is a pharmaceutical composition.

In one embodiment, the agent further comprises a second active ingredient. The second active ingredient may be selected from the group consisting of insulin, glucagon-like peptide-1 (GLP-1), biguanides, forskolin compounds, sulfonylureas, dipeptidyl peptidase-4 (DPP4) inhibitors, alpha-glucosidase inhibitors, thiazolidinediones, meglitinide and sodium-glucose cotransporter-2 (SGLT2) inhibitors.

Other methods

in one aspect, the invention relates to a method of treating an endocrine, nutritional, and/or metabolic disease comprising administering to a subject in need thereof an agent described herein.

in one aspect, the invention relates to the use of an agent for the manufacture of a medicament for the treatment of endocrine, nutritional and/or metabolic disorders in a mammal.

in one aspect, the invention relates to polynucleotides that, when expressed, encode a peptide as described herein. In one aspect, the invention relates to a vector comprising said polynucleotide encoding a peptide as described herein when expressed. In one aspect, the invention relates to a cell comprising said polynucleotide or said vector which encodes, when expressed, a peptide as described herein.

in one aspect, the invention relates to a method for increasing insulin secretion comprising administering to an individual in need thereof a therapeutically effective amount of a peptide described herein. In one embodiment, the method is an in vitro method.

in one aspect, the invention relates to a method for reducing blood glucose levels comprising administering to an individual in need thereof a therapeutically effective amount of a peptide described herein. In one embodiment, the method is an in vitro method. In one embodiment, insulin secretion is increased. In another embodiment, the uptake of glucose by the cell is increased. In another embodiment, insulin production is increased. In another embodiment, glucagon production is decreased.

In one aspect, the invention relates to a method, e.g., an in vitro method, for improving beta-cell morphology comprising administering to an individual in need thereof a therapeutically effective amount of a peptide described herein.

In one aspect, the invention relates to a method for improving beta-cell viability comprising administering to an individual in need thereof a therapeutically effective amount of a peptide described herein.

In one aspect, the invention relates to methods of delaying the onset of diabetes and diabetes-related conditions and diseases comprising administering to an individual in need thereof a therapeutically effective amount of a peptide described herein.

In one embodiment of the invention, the agent may further comprise a detectable moiety. For example, the detectable moiety may comprise or consist of a radioisotope, wherein, for example, the radioisotope is selected from the group consisting of 99mTc, 111In, 67Ga, 68Ga, 72As, 89Zr, 123I and 201 Tl. Thus, the conjugated moiety may be attached to a nanoparticle with multiple imaging capabilities (e.g., SPECT, PET, MRI, optical or ultrasound). Alternatively, the detectable moiety may comprise or consist of a paramagnetic isotope, wherein, for example, the paramagnetic isotope is selected from the group consisting of 157Gd, 55Mn, 162Dy, 52Cr, and 56 Fe.

where the agent comprises a detectable moiety, the detectable moiety may be detectable by imaging techniques (e.g., SPECT, PET, MRI, optical or ultrasound imaging).

In one aspect, the invention relates to the use of an agent as described herein for the preparation of a diagnostic composition for diagnosing a disease, disorder or injury of the pancreas in an individual.

Item(s)

1. an agent, comprising:

a) A peptide, wherein said peptide or said peptide analog comprises an amino acid sequence of the general formula:

KXLAXXXXIXLXYGIK(SEQ ID NO:140)

Wherein:

x2 is C, P or G;

X5 is E or G;

X6 is C, D or I;

x7 is D, I, S or G;

X8 is S, D or G;

X10 is E or G;

X12 is S or T;

provided that if X12 is T, then the peptide comprises no more than 25 amino acid residues; and

provided that if X2 is P, X5 is E, X6 is I, X7 is D, X8 is S, X10 is E, and X12 is S, then the peptide comprises no more than 85 amino acid residues;

Or a biologically active fragment and/or variant of SEQ ID NO. 140;

b) A polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c).

2. An agent comprising a peptide, wherein the peptide comprises an amino acid sequence of the general formula:

VDVPZGDISLAYZLR(SEQ ID NO:164)

Wherein:

z5 is E or N;

Z13 is R or G.

3. an agent comprising a peptide, wherein the peptide comprises an amino acid sequence of the general formula:

VDTYDGZZSVVYGLR(SEQ ID NO:165)

Wherein the content of the first and second substances,

z7 is D or G;

z8 is I or G.

4. An agent comprising a peptide, wherein the peptide comprises an amino acid sequence of the general formula:

GDPNZZZZZSVVYGLR(SEQ ID NO:166)

wherein the content of the first and second substances,

z5 is D or G;

Z6 is D or G;

Z7 is I or R;

Z8 is G or absent;

Z9 is D or absent.

5. the agent of items 2-4, wherein the agent comprises:

a) a peptide, wherein the peptide comprises an amino acid sequence selected from the group consisting of: GDPNDGRGDSVVYGLR (SEQ ID NO:137), VDTYDGGISVVYGLR (SEQ ID NO:138), and VDTYDGDGSVVYGLR (SEQ ID NO:139), VDVPEGDISLAYGLR (SEQ ID NO:157), LDGLVRAYDNISPVG (SEQ ID NO:158), GDPNGDISVVYGLR (SEQ ID NO:159), VDVPNGDISLAYRLR (SEQ ID NO:160), VDVPEGDISLAYRLR (SEQ ID NO:161), or consist thereof;

b) A polynucleotide encoding the peptide of a) upon expression;

c) A vector comprising the polynucleotide of b); and

d) a cell comprising the polynucleotide of b) or the vector of c).

6. The agent according to item 1, wherein the agent comprises or consists of an amino acid sequence of the general formula:

KXLAXXXXIXLSYGIK(SEQ ID NO:162)

Wherein the content of the first and second substances,

X2 is C, P or G;

X5 is E or G;

X6 is C, I or absent;

X7 is D, G or absent;

x8 is S, G or absent;

x10 is E or G.

7. The agent according to item 6, wherein the agent comprises or consists of an amino acid sequence of the general formula:

KXLAXIXLSYGIK(SEQ ID NO:163)

wherein the content of the first and second substances,

X2 is C, P or G;

X5 is E or G;

X10 is E or G.

8. An agent, comprising:

a) A peptide or peptide analog comprising or consisting of the amino acid sequences GDPNDGRGDSVVYGLR (SEQ ID NO:137), VDTYDGGISVVYGLR (SEQ ID NO:138), and VDTYDGDGSVVYGLR (SEQ ID NO:139), VDVPEGDISLAYGLR (SEQ ID NO:157), LDGLVRAYDNISPVG (SEQ ID NO:158), GDPNGDISVVYGLR (SEQ ID NO:159), VDVPNGDISLAYRLR (SEQ ID NO:160), VDVPEGDISLAYRLR (SEQ ID NO:161), V (. beta. -D) TYDGDISVVYGLR (SEQ ID NO:167), VDTY (. beta. -D) GDISVVYGLR (SEQ ID NO:168), VDTYDG (. beta. -D) ISVYGLR (SEQ ID NO: 169);

b) A polynucleotide encoding the peptide of a) upon expression;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c).

9. the agent of any one of the preceding claims, wherein the agent comprises non-naturally occurring (e.g., non-proteinogenic amino acid) amino acid residues.

10. An agent according to any preceding claim wherein the agent is conjugated to a moiety.

11. the agent according to any one of the preceding claims, wherein the moiety is selected from the group consisting of polyethylene glycol (PEG), monosaccharides, fluorophores, chromophores, radioactive compounds and cell penetrating peptides.

12. an agent according to any preceding claim, wherein the agent is further modified, for example by glycosylation, or by pegylation, amidation, esterification, acylation, acetylation, and/or alkylation.

13. The agent according to any one of the preceding claims, wherein the agent comprises or consists of a tandem repeat sequence.

14. The agent according to any one of the preceding claims wherein the tandem repeat sequence comprises or consists of the amino acid sequence of any one or more of the sequences described in the preceding items.

15. An agent according to any preceding claim, wherein the agent is fused to another polypeptide.

16. An agent according to any preceding claim, wherein the polypeptide is selected from the group consisting of glutathione-S-transferase (GST) and protein A.

17. The agent according to any one of the preceding claims, wherein the agent is fused to a tag.

18. The agent of any one of the preceding claims, wherein the tag is an oligo-histidine tag.

19. the agent according to any one of the preceding claims, wherein the agent is cyclic, for example wherein the peptide is cyclic.

20. An agent according to any preceding claim, wherein the peptide or peptide analogue is capable of forming at least one intramolecular cysteine bridge, for example, to form a cyclic or partially cyclic peptide.

21. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises an amino acid sequence selected from the group consisting of: KCLAECDSIELSYGIK (SEQ ID NO:141), CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEDISIELSYGIK (SEQ ID NO:145), KPLAEIGDIELSYGIK (SEQ ID NO:146), KPLAEGDIELSYGIK (SEQ ID NO:147), KPLAEIELSYGIK (SEQ ID NO:148), KPLAEIDSIELTYGIK (SEQ ID NO:149), KPLAEIDGIELSYGIK (SEQ ID NO:150), KPLAEIDGIELTYGIK (SEQ ID NO:151), KPLAEIGSIELSYGIK (SEQ ID NO:152), KGLAEIDSIELSYGIK (SEQ ID NO:153), KPLAGIDSIGLSYGIK (SEQ ID NO:154), KCLAEIDSCELSYGIK (SEQ ID NO:155) and CFKPLAEIDSIEC (SEQ ID NO: 156).

22. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAEIDSIELSYGIK (SEQ ID NO:136), or a variant or fragment thereof.

23. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAGIDSIGLSYGIK (SEQ ID NO:154), or a variant or fragment thereof.

24. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KGLAEIDSIELSYGIK (SEQ ID NO:153), or a variant or fragment thereof.

25. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KCLAECDSIELSYGIK (SEQ ID NO:141), or a variant or fragment thereof.

26. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAEIDGIELTYGIK (SEQ ID NO:151), or a variant or fragment thereof.

27. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAEIGSIELSYGIK (SEQ ID NO:152), or a variant or fragment thereof.

28. An agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of amino acid sequence KPLAEIELSYGIK (SEQ ID NO:148), or a variant or fragment thereof.

29. An agent according to any one of the preceding claims wherein the variant comprises or consists of a sequence in which any one amino acid has been changed to another proteinogenic amino acid or to a non-proteinogenic amino acid, provided that no more than 5 amino acids have been so changed.

30. an agent according to any preceding claim, wherein the variant comprises or consists of a sequence in which no more than 5 amino acids (e.g. no more than 4 amino acids, such as no more than 3 amino acids, such as no more than 2 amino acids, such as no more than 1 amino acid) have been changed to another proteinogenic or non-proteinogenic amino acid.

31. an agent according to any preceding claim wherein one or more amino acids are conservatively substituted.

32. an agent according to any one of the preceding claims, wherein the peptide or peptide analogue comprises or consists of one or more additional amino acids inserted at the N-and/or C-terminus and/or interior of the sequence.

33. An agent according to any preceding claim, wherein the peptide or peptide analogue comprises 1 additional amino acid conjugated to the N-or C-terminus.

34. the agent according to any one of the preceding claims, wherein the agent comprises no more than 85, such as no more than 80, such as no more than 75, such as no more than 70, such as no more than 65, such as no more than 60, such as no more than 55, such as no more than 50, such as no more than 55, such as no more than 40 amino acids, such as no more than 35, such as no more than 30, such as no more than 28, such as no more than 26, such as no more than 24, such as no more than 22, such as no more than 20, such as no more than 19, such as no more than 18, such as no more than 17, such as no more than 16, such as no more than 15, such as no more than 14, such as no more than 13, such as no more than 12, such as no more than 11, such as no.

35. An agent according to any one of the preceding claims, wherein the agent comprises at least 2 additional amino acids, such as at least 3, such as at least 4, such as at least 5, such as at least 6, such as at least 7, such as at least 8, such as at least 9, such as at least 10, such as at least 15, or such as at least 20 amino acids, conjugated to the N-or C-terminus of the peptide.

36. The agent according to any one of the preceding claims, wherein the agent further comprises a detectable moiety.

37. The agent of any one of the preceding claims, wherein the detectable moiety comprises or consists of a radioisotope.

38. The agent according to any one of the preceding claims, wherein the radioisotope is selected from the group consisting of 99mTc, 111In, 67Ga, 68Ga, 72As, 89Zr, 123I and 201 Tl.

39. An agent according to any preceding claim wherein the detectable moiety is detectable by an imaging technique (e.g. SPECT, PET, MRI, optical or ultrasound imaging).

40. Use of an agent according to any preceding claim in the preparation of a diagnostic composition for diagnosing a pancreatic disease, disorder or injury in an individual.

41. A composition comprising an agent according to any preceding claim.

42. The composition of any one of the preceding claims, wherein the composition is a pharmaceutical composition.

43. An agent or composition according to any one of the preceding claims for use as a medicament.

44. An agent selected from the group consisting of:

a) a peptide selected from the group consisting of:

A peptide comprising or consisting of an amino acid sequence of the general formula:

KXLAXXXXIXLXYGIK(SEQ ID NO:140)

Wherein:

x2 is C, P or G;

X5 is E or G;

x6 is C, D or I;

X7 is D, I, S or G;

X8 is S, D or G;

x10 is E or G;

X12 is S or T;

Provided that if X12 is T, then the peptide comprises no more than 25 amino acid residues;

or SEQ ID NO: 140;

(ii) a peptide comprising or consisting of an amino acid sequence of the general formula:

VDZZZGZZSZZYGLR(SEQ ID NO:68)

Wherein:

Z3 is T or V;

z4 is Y or P;

z5 is D or N;

Z7 is D or G;

z8 is I or G;

Z10 is V or L;

Z11is V or A; and

b) a polynucleotide encoding the peptide of a) when expressed;

c) A vector comprising the polynucleotide of b); and

d) A cell comprising the polynucleotide of b) or the vector of c);

for use in the treatment of endocrine, nutritional and/or metabolic disorders in a mammal.

45. The agent or composition for use of item 44, wherein the peptide comprises an amino acid sequence selected from the group consisting of: KCLAECDSIELSYGIK (SEQ ID NO:141), CLAEIDSC (SEQ ID NO:142), CFKPLAEIDSIECSYGIK (SEQ ID NO:143), KPLAEIELSYGIK (SEQ ID NO:148), KCLAEIDSCELSYGIK (SEQ ID NO:155) and CFKPLAEIDSIEC (SEQ ID NO:156), or consists thereof.

46. The agent or composition for use according to any one of the preceding claims, wherein the peptide is selected from the group consisting of SEQ ID NO: 141. 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, and 156.

47. The agent or composition for use according to any one of the preceding claims, wherein the peptide is selected from the group consisting of SEQ ID NO: 1. 136, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 67, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 115, 112,113, 125, 116, 125, 124, 121, 124, 126, 121, 124, 126, 121, 124, 23, 131. 132, 133, 135, 137, 138, 139, 157, 158, 159, 160, 161, 167, 168, and 169.

48. An agent or composition for use according to any preceding claim, wherein the agent comprises a second or additional active ingredient.

49. The agent or composition for use of item 48, wherein the second or additional active ingredient is selected from the group consisting of insulin, glucagon-like peptide-1 (GLP-1), sulfonylureas, dipeptidyl peptidase-4 (DPP4) inhibitors, alpha-glucosidase inhibitors, thiazolidinediones, meglitinide, and sodium-glucose cotransporter-2 (SGLT2) inhibitors.

50. Use of an agent or composition according to any one of the preceding claims for the treatment of an endocrine, nutritional and/or metabolic disorder in a mammal.

51. the agent or composition for use according to item 50, wherein the mammal is a human.

52. The agent or composition for use according to any one of the preceding claims, wherein the endocrine, nutritional and/or metabolic disease is selected from the group consisting of diabetes, type 1 diabetes, type 2 diabetes, diabetes related to malnutrition, glucose regulation and pancreatic endocrine disorders, insulin resistance syndrome, impaired glucose tolerance, hyperglycemia, hyperinsulinemia, and any combination thereof.

53. The agent or composition for use according to any one of the preceding claims, wherein the endocrine, nutritional and/or metabolic disease is selected from the group consisting of diabetes, thyroid disorders, glucose regulation and pancreatic endocrine disorders, endocrine gland disorders, nutritional deficiencies, obesity, overnutrition and metabolic disorders.

54. the agent or composition for use according to any one of the preceding claims, wherein the diabetes is selected from the group consisting of type 1 diabetes, type 2 diabetes, diabetes related to nutritional disorders, explicit diabetes and non-explicit diabetes.

55. The agent or composition for use according to any one of the preceding claims, wherein the glucose regulation and pancreatic endocrine disorder is selected from the group consisting of non-diabetic hypoglycaemic coma and pancreatic endocrine disorder.

56. The agent or composition for use according to any of the preceding claims, wherein the condition of obesity and overnutrition is selected from the group consisting of local obesity, overnutrition, and sequelae of overnutrition.

57. The agent or composition for use according to any of the preceding claims, wherein the nutritional deficiency is selected from the group consisting of disorders of aromatic amino acid metabolism, disorders of branched chain amino acid metabolism and fatty acid metabolism, disorders of amino acid metabolism, lactose intolerance, disorders of carbohydrate metabolism, disorders of sphingolipid metabolism, disorders of lipid storage, disorders of glycosaminoglycan metabolism, disorders of glycoprotein metabolism, disorders of lipoprotein metabolism, lipemia (lipiemia), disorders of purine and pyrimidine metabolism, disorders of porphyrin and bilirubin metabolism, disorders of mineral metabolism, cystic fibrosis, amyloidosis, hypovolemia, disorders of body fluids, electrolytes and acid-base balance, and post-operative endocrine and metabolic disorders.

58. a method of treating an endocrine, nutritional, and/or metabolic disease, the method comprising administering to a subject in need thereof an agent according to any one of the preceding.

59. Use of an agent according to any one of the preceding claims in the manufacture of a medicament for the treatment of an endocrine, nutritional and/or metabolic disorder in a mammal.

60. a method of delaying the onset of diabetes and diabetes-related conditions and diseases comprising administering to a subject in need thereof a therapeutically effective amount of an agent as defined in any one of the preceding claims.

61. A method for lowering blood glucose levels, comprising administering to an individual in need thereof a therapeutically effective amount of the agent of any one of the preceding claims.

62. The method of item 61, wherein secretion of insulin is increased.

63. the method of item 61, wherein uptake of glucose by the cell is increased.

64. The method of item 61, wherein insulin production is increased.

65. The method of item 61, wherein glucagon production is decreased.

66. a method of improving beta cell viability, comprising administering to a subject in need thereof a therapeutically effective amount of the agent of any one of the preceding claims.

67. A method of improving beta cell morphology comprising administering to an individual in need thereof a therapeutically effective amount of an agent of any one of the preceding claims.

68. A method of stabilizing or improving islet viability and/or morphology, comprising administering to an individual in need thereof a therapeutically effective amount of the agent of any one of the preceding claims.