GLUCOSE-REGULATING POLYPEPTIDES AND METHODS OF MAKING AND USING SAME

• US 20150274800A1
• Filed: 12/05/2014
• Published: 10/01/2015
• Est. Priority Date: 06/08/2009
• Status: Active Grant

• Current Assignee: Amunix Pharmaceuticals, Inc. (Sanofi )
• Original Assignee: Amunix Operating Inc. (Sanofi )
• Inventors: Schellenberger, Volker, Silverman, Joshua, Stemmer, Willem P., Wang, Chia-wei, Geething, Nathan, Cleland, Jeffrey L.
• Granted Patent: US 9,540,430 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A23V 2200/328 having effect on glycaemic control and diabetes
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/12 Antidiarrhoeals
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/00 Drugs for disorders of the urinary system
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 14/575 : Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 14/57563 : Vasoactive intestinal peptide [VIP]; Related peptides
• CPC Class Codes: C07K 14/605 : Glucagons
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/31 : fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/50 : containing protease site
• CPC Class Codes: C12N 15/70 : Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12P 21/02 : having a known sequence of two or more amino acids, e.g. glutathione

Abstract

The present invention relates to compositions comprising glucose regulating peptides linked to extended recombinant polypeptide (XTEN), isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of making and using such compositions in treatment of glucose regulating peptide-related diseases, disorders, and conditions.

Claims

1. A method of producing a fusion protein comprising a glucose regulating peptide that is at least 90% identical to an amino acid sequence selected from SEQ ID NO:

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, or SEQ ID NO;

10, wherein said glucose regulating peptide is linked to an extended recombinant polypeptide (XTEN), wherein the XTEN sequence exhibits at least 90% sequence identity to a comparable length of an amino acid sequence selected from the group consisting of the sequences of SEQ ID NOS;

189-207, comprising;

(a) providing host cell comprising a recombinant polynucleotide molecule encoding the fusion protein;

(b) culturing the host cell under conditions permitting the expression of the fusion protein; and

(c) recovering the fusion protein.

2. The method of claim 1, wherein the glucose regulating peptide of the fusion protein is at least 95% identical to an amino acid sequence selected from SEQ ID NO:

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, or SEQ ID NO;

10.

3. The method of claim 1, wherein the glucose regulating peptide of the fusion protein is a sequence selected from SEQ ID NO:

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, or SEQ ID NO;

10.

4. The method of claim 1, wherein the glucose regulating peptide of the fusion protein is at least 90% identical to the amino acid sequence SEQ ID NO:

4.

5. The method of claim 1, wherein the glucose regulating peptide of the fusion protein is at least 90% identical to the amino acid sequence SEQ ID NO:

5.

6. The method of claim 1, wherein the glucose regulating peptide of the fusion protein is at least 90% identical to the amino acid sequence SEQ ID NO:

6.

7. The method of claim 1, wherein the glucose regulating peptide of the fusion protein is at least 90% identical to the amino acid sequence SEQ ID NO:

10.

8. The method of claim 1, wherein the XTEN sequence exhibits at least 90% sequence identity to a comparable length of an amino acid sequence selected from SEQ ID NO:

189, SEQ ID NO;

191, or SEQ ID NO;

199.

9. The method of claim 1, wherein the polynucleotide is codon optimized for enhanced expression of said fusion protein in the host cell.

10. The method of claim 9, wherein the host cell is a prokaryotic cell.

11. The method of claim 1, wherein the isolated fusion protein is recovered from the host cell cytoplasm in substantially soluble form.

12. An isolated nucleic acid comprising a nucleotide sequence encoding a fusion protein comprising a glucose regulating peptide that is at least 90% identical to an amino acid sequence selected from SEQ ID NO:

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, or SEQ ID NO;

10, wherein said glucose regulating peptide is linked to an extended recombinant polypeptide (XTEN), wherein the XTEN sequence exhibits at least 90% sequence identity to a comparable length of an amino acid sequence selected from the group consisting of the sequences of SEQ ID NOS;

189-207.

13. An expression vector comprising the nucleic acid of claim 12.

14. An isolated host cell comprising the nucleic acid of claim 12, wherein the host cell is a prokaryotic host cell.

15. A method of treating a glucose regulating peptide related condition in a subject, comprising administering to the subject a therapeutically effective amount of a fusion protein comprising a glucose regulating peptide that is at least 90% identical to an amino acid sequence selected from SEQ ID NO:

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, or SEQ ID NO;

10, wherein said glucose regulating peptide is linked to an extended recombinant polypeptide (XTEN), wherein the XTEN sequence exhibits at least 90% sequence identity to a comparable length of an amino acid sequence selected from the group consisting of the sequences of SEQ ID NOS;

189-207.

16. The method of claim 15, wherein the glucose regulating peptide related condition is selected from juvenile diabetes, type I diabetes, type II diabetes, obesity, acute hypoglycemia, acute hyperglycemia, nocturnal hypoglycemia, chronic hyperglycemia, glucagonomas, secretory disorders of the airway, arthritis, osteoporosis, central nervous system disease, restenosis, neurodegenerative disease, renal failure, congestive heart failure, nephrotic syndrome, cirrhosis, pulmonary edema, hypertension, stroke, irritable bowel syndrome, myocardial infarction, acute coronary syndrome, post-surgical catabolic changes, hibernating myocardium, diabetic cardiomyopathy, insufficient urinary sodium excretion, excessive urinary potassium concentration, polycystic ovary syndrome, respiratory distress, nephropathy, left ventricular systolic dysfunction, diarrhea, postoperative dumping syndrome, critical illness polyneuropathy (CIPN), dyslipidemia, organ tissue injury caused by reperfusion of blood flow following ischemia, and coronary heart disease risk factor (CHDRF) syndrome.

17. The method of claim 15, wherein the therapeutically effective amount results in maintaining blood concentrations of the fusion protein within a therapeutic window for the fusion protein at least three-fold longer compared to the corresponding native glucose regulating peptide that lacks the XTEN administered at a comparable amount to a subject.

18. The method of claim 15, wherein administration of two or more consecutive doses of the fusion protein administered using a therapeutically effective dose regimen to a subject results in a gain in time between consecutive C_max peaks and/or C_min troughs for blood levels of the fusion protein compared to the corresponding glucose regulating peptide not linked to the fusion protein and administered to a subject using a therapeutically effective dose regimen established for the glucose regulating peptide.

19. The method of claim 15, wherein

(i) a smaller molar amount of the fusion protein is administered to a subject in comparison to the corresponding glucose regulating peptide that lacks the XTEN administered to a subject under an otherwise equivalent dose regimen, and the fusion protein achieves a comparable therapeutic effect as the corresponding glucose regulating peptide that lacks the XTEN;

(ii) the fusion protein is administered less frequently to a subject in comparison to the corresponding glucose regulating peptide that lacks the XTEN administered to a subject using an otherwise equivalent molar dose, and the fusion protein achieves a comparable therapeutic effect as the corresponding glucose regulating peptide that lacks the XTEN;

or

(iii) an accumulatively smaller molar amount of the fusion protein is administered to a subject in comparison to the corresponding glucose regulating peptide that lacks the XTEN administered to a subject under the otherwise equivalent dose period, the fusion protein achieves a comparable therapeutic effect as the corresponding glucose regulating peptide that lacks the XTEN.

20. The method of claim 19, wherein the therapeutic effect is a measured parameter selected from HbAlc concentrations, insulin concentrations, stimulated C peptide, fasting plasma glucose (FPG), serum cytokine levels, CRP levels, insulin secretion and Insulin-sensitivity index derived from an oral glucose tolerance test (OGTT), body weight, and food consumption.