CONJUGATE COMPRISING OXYNTOMODULIN AND AN IMMUNOGLOBULIN FRAGMENT, AND USE THEREOF

• US 20140212440A1
• Filed: 06/15/2012
• Published: 07/31/2014
• Est. Priority Date: 06/17/2011
• Status: Active Grant

• Current Assignee: Hanmi Science Co., Ltd
• Original Assignee: Hanmi Science Co., Ltd
• Inventors: Jung, Sung Youb, Kim, Dae Jin, Park, Sung Hee, Woo, Young Eun, Choi, In Young, Kwon, Se Chang
• Granted Patent: US 9,731,031 B2
• Time in Patent Office: Days
• US Class Current: 424/178.1
• 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: A61K 38/26 Glucagons
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/68 the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
• CPC Class Codes: A61K 47/6801 Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
• CPC Class Codes: A61K 47/6811 the drug being a protein or peptide, e.g. transferrin or bleomycin
• CPC Class Codes: A61K 47/6889 Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• 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 14/605 Glucagons
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The present invention relates to a conjugate comprising oxyntomodulin, an immunoglobulin Fc region, and non-peptidyl polymer wherein the conjugate being obtainable by covalently linking oxyntomodulin to immunoglobulin Fc region via non-peptidyl polymer, and a pharmaceutical composition for the prevention or treatment of obesity comprising the conjugates. The conjugate comprising oxyntomodulin and the immunoglobulin Fc of the present invention reduces food intake, suppresses gastric emptying, and facilitates lipolysis without side-effects, unlike native oxyntomodulin, and also shows excellent receptor-activating effects and long-term sustainability, compared to native oxyntomodulin. Thus, it can be widely used in the treatment of obesity with safety and efficacy.

Claims

1. A conjugate comprising oxyntomodulin, an immunoglobulin Fc region, and non-peptidyl polymer wherein the conjugate being obtainable by covalently linking oxyntomodulin to immunoglobulin Fc region via non-peptidyl polymer.

2. The conjugate according to claim 1, wherein the oxyntomodulin has the amino acid sequence of SEQ ID NO. 1.

3. The conjugate according to claim 1, wherein the oxyntomodulin is an oxyntomodulin derivative prepared by addition, deletion or substitution of amino acids of native oxyntomodulin.

4. The conjugate according to claim 3, wherein the oxyntomodulin derivative comprising the amino acid sequence of following Formula 1:

R1-X1-X2-GTFTSD-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-X24-R2 

 

(Formula

     1)

wherein R1 is histidine, desamino-histidyl, dimethyl-histidyl (N-dimethyl-histidyl), beta-hydroxyimidazopropionyl, 4-imidazoacetyl, beta-carboxy imidazopropionyl or tyrosine;

X1 is Aib(aminosiobutyric acid), d-alanine, glycine, Sar(N-methylglycine), serine, or d-serine;

X2 is glutamic acid or glutamine;

X3 is leucine or tyrosine;

X4 is serine or alanine;

X5 is lysine or arginine;

X6 is glutamine or tyrosine;

X7 is leucine or methionine;

X8 is aspartic acid or glutamic acid;

X9 is glutamic acid, serine, alpha-methyl-glutamic acid or is deleted;

X10 is glutamine, glutamic acid, lysine, arginine, serine or is deleted;

X11 is alanine, arginine, valine or is deleted;

X12 is alanine, arginine, serine, valine or is deleted;

X13 is lysine, glutamine, arginine, alpha-methyl-glutamic acid or is deleted;

X14 is aspartic acid, glutamic acid, leucine or is deleted;

X15 is phenylalanine or is deleted;

X16 is isoleucine, valine or is deleted;

X17 is alanine, cysteine, glutamic acid, lysine, glutamine, alpha-methyl-glutamic acid or is deleted;

X18 is tryptophan or is deleted;

X19 is alanine, isoleucine, leucine, serine, valine or is deleted;

X20 is alanine, lysine, methionine, glutamine, arginine or is deleted;

X21 is asparagine or is deleted;

X22 is alanine, glycine, threonine or is deleted;

X23 is cysteine, lysine or is deleted;

X24 is a peptide having 2 to 10 amino acids consisting of combinations of alanine, glycine and serine, or is deleted; and

R2 is KRNRNNIA (SEQ ID NO.

     35), GPSSGAPPPS (SEQ ID NO.

     36), GPSSGAPPPSK (SEQ ID NO.

     37), HSQGTFTSDYSKYLD (SEQ ID NO.

     38), HSQGTFTSDYSRYLDK (SEQ ID NO.

     39), HGEGTFTSDLSKQMEEEAVK (SEQ ID NO.

     40) or is deleted (excluded if the amino acid sequence of Formula 1 is identical to that of SEQ ID NO.

     1).

5. The conjugate according to claim 4, wherein the oxyntomodulin derivative is capable of activating GLP-1 receptor and glucagon receptor.

6. The conjugate according to claim 4, wherein the conjugate has anti-obesity effects.

7. The conjugate according to claim 4, wherein one or more amino acids at positions 10, 14, 16, 20, 24 and 28 of the amino acid sequence of Formula 1 are substituted with amino acids or amino acid derivatives selected from the group consisting of Tyr(4-Me), Phe, Phe(4-Me), Phe(4-Cl), Phe(4-CN), Phe(4-NO₂), Phe(4-NH2), Phg, Pal, NaI, Ala(2-thienyl) and Ala(benzothienyl).

8. The conjugate according to claim 4, wherein one or more amino acid pairs at positions 10 and 14, 12 and 16, 16 and 20, 20 and 24, and 24 and 28 of the amino acid sequence of Formula 1 are substituted with glutamic acid or lysine to form rings.

9. The conjugate according to claim 4, wherein one or more amino acid pairs at positions 10 and 14, 12 and 16, 16 and 20, 20 and 24, and 24 and 28 of the amino acid sequence of Formula 1 form rings.

10. The conjugate according to claim 4, wherein the oxyntomodulin derivative is selected from the group consisting of the peptides of SEQ ID NOs. 2 to 34.

11. The conjugate according to claim 1, wherein the non-peptidyl polymer is selected from the group consisting of polyethylene glycol, polypropylene glycol, copolymers of ethylene glycol and propylene glycol, polyoxyethylated polyols, polyvinyl alcohol, polysaccharides, dextran, polyvinyl ethyl ether, polylactic acid (PLA), polylactic-glycolic acid (PLGA), lipid polymers, chitins, hyaluronic acid, polysaccharide and combinations thereof.

12. The conjugate according to claim 1, wherein an amine group and a thiol group of the immunoglobulin Fc region and oxyntomodulin are linked at both ends of the non-peptidyl polymer, respectively.

13. The conjugate according to claim 1, wherein the non-peptidyl polymer has reactive groups capable of binding with the immunoglobulin Fc region and the protein drug at both ends.

14. The conjugate according to claim 13, wherein the reactive group is selected from the group consisting of an aldehyde group, a propionaldehyde group, a butyraldehyde group, a maleimide group and a succinimide derivative.

15. The conjugate according to claim 13, wherein the reactive groups at both ends are the same as or different from each other.

16. The conjugate according to claim 1, wherein the immunoglobulin Fc region is a non-glycosylated Fc region.

17. The conjugate according to claim 1, wherein the immunoglobulin Fc region is selected from the group consisting of a CH1 domain, a CH2 domain, a CH3 domain and a CH4 domain;

a CH1 domain and a CH2 domain;

a CH1 domain and a CH3 domain;

a CH2 domain and a CH3 domain;

a combination of one or more domains and an immunoglobulin hinge region (or a portion of the hinge region); and

a dimer of each domain of the heavy-chain constant regions and the light-chain constant region.

18. The conjugate according to claim 1, wherein the immunoglobulin Fc region is a derivative in which a region capable of forming a disulfide bond is deleted, certain amino acid residues are eliminated at the N-terminal end of a native Fc form, a methionine residue is added at the N-terminal end of a native Fc form, a complement-binding site is deleted, or an antibody dependent cell mediated cytotoxicity (ADCC) site is deleted.

19. The conjugate according to claim 1, wherein the immunoglobulin Fc region is an Fc region derived from an immunoglobulin selected from the group consisting of IgG, IgA, IgD, IgE, and IgM.

20. The conjugate according to claim 19, wherein the immunoglobulin Fc region is an IgG4 Fc region.

21. The conjugate according to claim 1, wherein the immunoglobulin Fc region is a human IgG4-derived non-glycosylated Fc region.

22. A pharmaceutical composition for the prevention or treatment of obesity, comprising the conjugate of claim 1.

23. The pharmaceutical composition according to claim 22, further comprising a pharmaceutically acceptable carrier.

24. The pharmaceutical composition according to claim 22, wherein the composition is administered alone or in combination with other pharmaceutical formulations showing prophylactic or therapeutic effects on obesity.

25. The pharmaceutical composition according to claim 24, wherein the pharmaceutical formulation is selected from the group consisting of a GLP-1 receptor agonist, a leptin receptor agonist, a DPP-IV inhibitor, a Y5 receptor antagonist, a Melanin-concentrating hormone (MCH) receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an Amylin receptor agonist, a Ghrelin antagonist, and a Ghrelin receptor antagonist.

26. A method for preventing or treating obesity, comprising the step of administering the conjugate of claim 1 to a subject.

27. (canceled)