Fatty acids and their use in conjugation to biomolecules

Patent metadata

Field	Value
Publication number	US-10588980-B2
Application number	US-201514738272-A
Country	US
Kind code	B2
Filing date	Jun 12, 2015
Priority date	Jun 23, 2014
Publication date	Mar 17, 2020
Grant date	Mar 17, 2020

Abstract

The invention provides a conjugate comprising a biomolecule linked to a fatty acid via a linker wherein the fatty acid has the following Formulae A1, A2 or A3:

wherein R 1 , R 2 , R 3 , R 4 , Ak, n, m and p are defined herein. The invention also relates to a method for manufacturing the conjugate of the invention such as GDF15 conjugate, and its therapeutic uses such as treatment or prevention of metabolic disorders or diseases, type 2 diabetes mellitus, obesity, pancreatitis, dyslipidemia, alcoholic and nonalcoholic fatty liver disease/steatohepatitis and other progressive liver diseases, insulin resistance, hyperinsulinemia, glucose intolerance, hyperglycemia, metabolic syndrome, hypertension, cardiovascular disease, atherosclerosis, peripheral arterial disease, stroke, heart failure, coronary heart disease, diabetic complications (including but not limited to chronic kidney disease), neuropathy, gastroparesis and other metabolic disorders. The present invention further provides a combination of pharmacologically active agents and a pharmaceutical composition.

First claim

What is claimed is:

1. A conjugate comprising a biomolecule linked to a fatty acid via a linker
wherein the fatty acid has the following Formulae A1:

wherin:
R 1 is CO 2 H;
R 2 , R 3 and are independently of each other H, OH, CO 2 H, —CH═CH 2 or —C≡CH; and
n and m are independently of each other an integer between 6 and 30; and
wherein the biomolecule is MH(199-308)hGDF15 (SEQ ID NO: 4), MHA(200-308)hGDF15 (SEQ ID NO: 6), AHA(200-308)hGDF15 (SEQ ID NO: 7), AH(199-308)hGDF15 (SEQ ID NO: 5), MHHHHHHM-hGDF15 (SEQ ID NO: 2), MHHHHHH-hGDF15 (SEQ ID NO: 1), or his-hGDF15; or a dimer thereof, wherein his-hGDF15 is hGDF15(197-308) wherein a tag, comprising 1 to 6 histidine amino acids and optionally 1 or 2 methionine amino acids, has been added to the N-terminus of hGDF15;
or an amide, ester, or pharmaceutically acceptable salt thereof.

2. The conjugate according to claim 1 , wherein the fatty acid is selected from:

wherein Ak 3 , Ak 4 , Ak 5 , Ak 6 and Ak 7 are independently a (C 8-20 )alkylene, and R 5 and R 6 are independently linear (C 8-20 )alkyl,
or an amide, an ester or a pharmaceutically acceptable salt thereof.

3. The conjugate according to claim 1 , wherein the fatty acid is selected from:

or an amide, ester, or a pharmaceutically acceptable salt thereof.

4. The conjugate according to claim 1 , wherein the linker comprises alkyl, alkenyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, polyethylene glycol, or one or more natural or unnatural amino acids, or combination thereof, wherein each of the alkyl, alkenyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, polyethylene glycol and/or the natural or unnatural amino acids are optionally combined and linked together or linked to the biomolecule and/or to the fatty acid moiety via a chemical group selected from —C(O)O—, —OC(O)—, —NHC(O)—, —C(O)NH—, —O—, —NH—, —S—, —C(O)—, —OC(O)NH—, —NHC(O)—O—, ═NH—O—, ═NH—NH — and ═NH—N(alkyl)-; or an amide, ester or a pharmaceutically acceptable salt thereof.

5. The conjugate according to claim 1 , wherein the linker comprises an unbranched oligo ethylene glycol moiety of Formula:

wherein y is 0 to 34; or an amide, an ester or a pharmaceutically acceptable salt thereof.

6. The conjugate according to claim 1 , wherein the linker comprises a heterocyclic moiety of the following Formula:

wherein r is an integer of 0 to 2 and s is an integer of 0 to 3;
or an amide, an ester or a pharmaceutically acceptable salt thereof.

7. The conjugate according to claim 1 , wherein the linker comprises one or more amino acids independently selected from histidine, methionine, alanine, glutamine, asparagine and glycine; or an amide, an ester or a pharmaceutically acceptable salt thereof.

8. A conjugate according to claim 1 , wherein the biomolecule linked to the fatty acid via a linker has one of the following Formulae:

wherein in Formulae C and D, both monomeric units of his-hGDF15 or of hGDF15* are linked to the fatty acid moiety via a linker at the N-terminus of each his-hGDF15 monomeric unit and at the N-terminus of each hGDF15* monomeric unit;
wherein in Formulae E and F, only one of the monomeric unit of his-hGDF15 or of hGDF15* is linked to the fatty acid moiety via a linker at the N-terminus; and
wherein in Formulae D and F, hGDF15* is MH(199-308)hGDF15 (SEQ ID NO:4), MHA(200-308)hGDF15 (SEQ ID NO:6), AHA(200-308)hGDF15 (SEQ ID NO:7), AH(199-308)hGDF15 (SEQ ID NO:5), MHHHHHH-hGDF15 (SEQ ID NO:2), or MHHHHHH-hGDF15 (SEQ ID NO:1);
wherein in Formulae C, D, E and F, s is an integer between 20-30; and
wherein in Formulae C, D, E and F, the line between the 2 monomeric units of his-hDGF15 or the 2 monomeric units of hGDF15* represent a disulfide bond.

9. A mixture comprising the conjugate according to claim 8 having Formula C and the conjugate according to claim 8 having Formula E or a mixture comprising the conjugate according to claim 8 having Formula D and the conjugate according to claim 8 having Formula F.

10. The conjugate according to claim 8 , wherein the biomolecule linked to a fatty acid via a linkers is Of Formula G or of Formula H:

wherein AHA-hGDF15 is AHA(200-308)hGDF15 (SEQ ID NO: 7)and the fatty acid is linked via a linker at the N-terminus of one AHA-hGDF15 monomeric unit in Formula H or via a linker at both the N-terminus of each of the of two AHA-hGDF15 monomeric units in Formula G, and wherein the line between the two AHA-hGDF15 units represent a disulfide bond.

11. A mixture comprising the conjugate according to claim 10 having Formula G and the conjugate according to claim 10 having Formula H.

12. The conjugate according to claim 1 , wherein the fatty acid moiety is attached the N-terminus of the biomolecule via a linker; or an amide, an ester or a pharmaceutically acceptable salt thereof.

13. The conjugate according to claim 1 , wherein the conjugate has a plasma stability half-life of more than 10 hours, more than 20 hours or more than 30 hours.

14. The conjugate according to claim 1 , where the improvement of plasma stability compared to the non-conjugated biomolecule is 2 fold, 5 fold, 10 fold, 20 fold, 30 fold, 40 fold, 50 fold, or 75 fold.

15. A combination comprising a therapeutically effective amount of a conjugate according to claim 1 , or an amide, an ester or a pharmaceutically acceptable salt thereof; and one or more therapeutically active co-agent.

16. The combination according to claim 15 , wherein the co-agent is selected from antidiabetic agent, hypolipidemic agent, anti-obesity agents, anti-hypertensive agents, and agonists of peroxisome proliferator-activator receptors.

17. The combination according to claim 16 , wherein the co-agent is selected from insulin, insulin derivatives and mimetics; insulin secretagogues; glyburide, glimepiridine; insulinotropic sulfonylurea receptor ligands; thiazolidinediones, pioglitazone, balaglitazone, rivoglitazone, netoglitazone, troglitazone, englitazone, ciglitazone, adaglitazone, darglitazone, Cholesteryl ester transfer protein (CETP) inhibitors, GSK3 (glycogen synthase kinase-3) inhibitors; Retinoid X receptor ligands; sodium-dependent glucose cotransporter inhibitors; glycogen phosphorylase A inhibitors; biguanides; alpha-glucosidase inhibitors, GLP-1 (glucagon like peptide-1), GLP-1 analogs, GLP-1 mimetics; DPPIV (dipeptidyl peptidase IV) inhibitors, 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitors; squalene synthase inhibitors; FXR (farnesoid X receptor), LXR (liver X receptor) ligands; cholestyramine; fibrates; nicotinic acid, aspirin; orlistat or rimonabant; l

Assignees

• Novartis Ag

Inventors

• Bruce Alexandra Marshall
• Usera Aimee Richardson
• Zecri Frederic
• Yuan Jun
• Lou Changgang
• Kanter Aaron

Classifications

• A61K38/095 Primary

  Oxytocins; Vasopressins; Related peptides · CPC title

• A61P9/14

  Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers · CPC title

• A61P9/12

  Antihypertensives · CPC title

• A61P9/10

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

• A61P9/04

  Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure · CPC title