Pituitary adenylate cyclase activating peptide (PACAP) receptor (VPAC2) agonists and their pharmacological methods of use

Abstract

This invention provides novel peptides that function in vivo as agonists of the VPAC2 receptor. These insulin secretagogue polypeptides are shown to lower blood glucose in vivo more than controls upon glucose challenge. The polypeptides of this invention are also stable in formulation and have long half-lives. The peptides of the present invention provide a new therapy for patients with decreased endogenous insulin secretion, in particular type 2 diabetics. In particular, the invention is a polypeptide selected from a specific group of VPAC2-related polypeptides, or functional equivalents thereof. The invention is also directed to a method of treating a metabolic disease in a mammal comprising administering a therapeutically effective amount of the insulin secretagogue peptides to said mammal. Also disclosed are methods of making the peptides, both recombinant and synthetic.

1 . A polypeptide having VPAC2 receptor agonist activity, selected from the group: consisting of SEQ ID NOs: 1 through 152, and functionally equivalent fragments, derivatives and variants thereof having at least 90% homology to the amino acid sequence of SEQ ID NO: 115. 
     
     
         2 . A polynucleotide encoding a polypeptide sequence of  claim 1 , or a degenerate variant thereof. 
     
     
         3 . A vector comprising a polynucleotide of  claim 2 . 
     
     
         4 . A host cell comprising a vector of  claim 3 . 
     
     
         5 . A method for producing a polypeptide comprising: a) culturing the host cell of  claim 4  under conditions suitable for the expression of said polypeptide; and b) recovering the polypeptide from the host cell culture. 
     
     
         6 . A purified antibody which binds specifically to the polypeptide of  claim 1 . 
     
     
         7 . The polypeptide of  claim 1 , wherein said polypeptide is selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, and 5. 
     
     
         8 . The polypeptide of  claim 1 , wherein said polypeptide is selected from the group consisting of SEQ ID NOs: 115, 116, 117, 118, and 119. 
     
     
         9 . A polynucleotide selected from the group consisting of SEQ ID NOs: 154 through 264. 
     
     
         10 . A pharmaceutical composition comprising a therapeutically effective amount of a polypeptide of  claim 1 , or functionally equivalent fragments, derivatives, and variants thereof, in combination with a pharmaceutically acceptable carrier. 
     
     
         11 . The pharmaceutical composition of  claim 10 , wherein said composition comprises about 2% to about 30% DMSO and optionally, a solvent selected from the consisting of propylene glycol, dimethyl formamide, propylene carbonate, polyethylene glycol, and triglycerides. 
     
     
         12 . The pharmaceutical composition of  claim 10 , wherein said polypeptide is selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 115, 116, 117, 118, and 119. 
     
     
         13 . A pharmaceutical composition comprising a therapeutically effective amount of a polypeptide of  claim 1 , or functionally equivalent fragments, derivatives, and variants thereof, in combination with a pharmaceutically acceptable carrier and one or more pharmaceutical agents. 
     
     
         14 . The pharmaceutical composition of  claim 13 , wherein said pharmaceutical agent is selected from the group consisting of PPAR agonists, sulfonylurea drugs, non-sulfonylurea secretagogues, α-glucosidase inhibitors, insulin sensitizers, insulin secretagogues, hepatic glucose output lowering compounds, insulin, anti-obesity agents, HMG CoA reductase inhibitors, nicotinic acid, bile acid sequestrants, fibric acid derivatives, and anti-hypertensive agents. 
     
     
         15 . A composition comprising an effective amount of a polypeptide of  claim 1 , or functionally equivalent fragments, derivatives, and variants thereof, in combination with an inert carrier. 
     
     
         16 . A method of treating diabetes comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         17 . The method of  claim 16 , wherein said diabetes is selected from the group consisting of type 1 diabetes, type 2 diabetes, maturity-onset diabetes of the young, latent autoimmune diabetes adult, and gestational diabetes. 
     
     
         18 . A method of treating Syndrome X comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         19 . A method of treating diabetes-related disorders comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         20 . The method of  claim 19 , wherein said diabetes-related disorder is selected from the group consisting of hyperglycemia, hyperinsulinemia, impaired glucose tolerance, impaired fasting glucose, dyslipidemia, hypertriglyceridemia, and insulin resistance. 
     
     
         21 . A method of treating diabetes comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1  in combination with one or more pharmaceutical agents. 
     
     
         22 . The method of  claim 21 , wherein said pharmaceutical agent is selected from the group consisting of PPAR agonists, sulfonylurea drugs, non-sulfonylurea secretagogues, α-glucosidase inhibitors, insulin sensitizers, insulin secretagogues, hepatic glucose output lowering compounds, insulin, and anti-obesity agents. 
     
     
         23 . The method of  claim 22 , wherein said diabetes is selected from the group consisting of type 1 diabetes, type 2 diabetes, maturity-onset diabetes of the young, latent autoimmune diabetes adult, and gestational diabetes. 
     
     
         24 . A method of treating Syndrome X comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1  in combination with one or more pharmaceutical agents. 
     
     
         25 . The method of  claim 24 , wherein said pharmaceutical agent is selected from the group consisting of PPAR agonists, sulfonylurea drugs, non-sulfonylurea secretagogues, α-glucosidase inhibitors, insulin sensitizers, insulin secretagogues, hepatic glucose output lowering compounds, insulin, and anti-obesity agents. 
     
     
         26 . A method of treating diabetes-related disorders comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1  in combination with one or more pharmaceutical agents. 
     
     
         27 . The method of  claim 26 , wherein said diabetes-related disorder is selected from the group consisting of hyperglycemia, hyperinsulinemia, impaired glucose tolerance, impaired fasting glucose, dyslipidemia, hypertriglyceridemia, and insulin resistance. 
     
     
         28 . The method of  claim 27 , wherein said pharmaceutical agent is selected from the group consisting of PPAR agonists, sulfonylurea drugs, non-sulfonylurea secretagogues, α-glucosidase inhibitors, insulin sensitizers, insulin secretagogues, hepatic glucose output lowering compounds, insulin, and anti-obesity agents. 
     
     
         29 . A method of treating diabetes, Syndrome X, or diabetes-related disorders comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1  in combination with one or more agents selected from the group consisting of HMG CoA reductase inhibitors, nicotinic acid, bile acid sequestrants, fibric acid derivatives, and anti-hypertensive agents. 
     
     
         30 . The method of  claim 29 , wherein said diabetes-related disorder is selected from the group consisting of hyperglycemia, hyperinsulinemia, impaired glucose tolerance, impaired fasting glucose, dyslipidemia, hypertriglyceridemia, and insulin resistance. 
     
     
         31 . The method of any one of  claims 21  to  30 , wherein the polypeptide of  claim 1  and one or more pharmaceutical agents are administered as a single pharmaceutical dosage formulation. 
     
     
         32 . A method of treating or preventing secondary causes of diabetes comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         33 . The method of  claim 32 , wherein said secondary cause is selected from the group consisting of glucocorticoid excess, growth hormone excess, pheochromocytoma, and drug-induced diabetes. 
     
     
         34 . A method of treating or preventing secondary causes of diabetes comprising the step of administering a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1  in combination with one or more pharmaceutical agents. 
     
     
         35 . The method of  claim 34 , wherein said pharmaceutical agent is selected from the group consisting of PPAR agonists, sulfonylurea drugs, non-sulfonylurea secretagogues, α-glucosidase inhibitors, insulin sensitizers, insulin secretagogues, hepatic glucose output lowering compounds, insulin, and anti-obesity agents. 
     
     
         36 . A method of treating respiratory disease comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         37 . A method of treating obesity comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         38 . A method of regulating appetite comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         39 . A method of treating cardiovascular disease comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         40 . A method of treating disorders of lipid and carbohydrate metabolism comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         41 . A method of treating sleep disorders comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         42 . A method of treating male reproductive disorders comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         43 . A method of treating growth disorders or disorders of energy homeostasis comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         44 . A method of treating immune diseases comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         45 . A method of treating autoimmune diseases comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         46 . A method of treating acute and chronic inflammatory diseases comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         47 . A method of treating septic shock comprising the step of administering to a subject in need thereof a therapeutically effective amount of a polypeptide of  claim 1 . 
     
     
         48 . A method of stimulating insulin release in a glucose-dependent manner in a subject in need thereof by administering to said subject a polypeptide of  claim 1 . 
     
     
         49 . A gene therapy composition comprising a polynucleotide of  claim 2  in combination with a therapeutically effective gene therapy vector.