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EP-4736944-A2 - POLYPEPTIDES AND USES THEREOF

EP4736944A2EP 4736944 A2EP4736944 A2EP 4736944A2EP-4736944-A2

Abstract

Disclosed are polypeptides which are pramlintide analogues and uses thereof. In particular, the present invention relates to polypeptides of SEQ ID NO 2 which are pramlintide analogues conjugated to half-life extending moieties such as albumin binding moieties and uses thereof.

Inventors

  • BEDNAREK, MARIA ALEKSANDRA
  • GENAPATHY, Sivaneswary

Assignees

  • MedImmune Limited

Dates

Publication Date
20260506
Application Date
20211215

Claims (15)

  1. A polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: wherein: Xaa (-4) is Lys(albumin binding moiety) or is absent; Xaa (-3) is Gly or is absent; Xaa (-2) is Gly or is absent; Xaa (-1) is Gly, (albumin binding moiety), Lys(albumin binding moiety) or is absent; Xaa 1 is Lys, Lys(albumin binding moiety), (albumin binding moiety) or is absent; Xaa 4 is Thr, Ile or Ala; Xaa 14 is Asn, His, Glu, 2,4-diaminobutanoic acid (Dab), or an alpha methyl amino acid; Xaa 15 is Phe or Trp; Xaa 16 is Leu or D -Leu (dL); Xaa 17 is Val, Ser, Glu, Arg, (2 S ,4 R )-4-hydroxypyrrolidine-2-carboxylic acid (Hyp), Dab or an alpha methyl amino acid ( e.g . 2-amino-2-methylpropanoic acid [Aib]); Xaa 20 is Ser, Ile, Pro or an alpha methyl amino acid ( e.g. (S) -2-amino-3-hydroxy-2-methylpropanoic acid [αMeSer]); Xaa 21 is Asn, Dab, His, Pro, Ser, Arg, Lys, Gly or Glu, Ala, Hyp or an alpha methyl amino acid ( e.g. Aib); Xaa 22 is Asn, His, Hyp, Dab or an alpha methyl amino acid ( e.g . Aib); Xaa 23 is Phe, Hyp or an alpha methyl amino acid ( e.g . (S) -2-amino-2-methyl-3-phenylpropanoic acid [αMePhe]); Xaa 24 is Gly, Pro, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 25 is Pro, Ala, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 26 is Ile, D -Ile (dl), Arg, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 27 is Leu, dL, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 28 is Pro, D -Pro (dP), Ser, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 29 is Pro, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 31 is Asn, Glu, His, Arg, Pro, Dab or an alpha methyl amino acid ( e.g . Aib); Xaa 32 is Val, Hyp, Dab or an alpha methyl amino acid ( e.g . Aib); Xaa 33 is Gly, Pro, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 34 is Ser, Pro, His, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 35 is Asn, Pro, Arg, Glu, Dab, Hyp or an alpha methyl amino acid ( e.g . Aib); Xaa 36 is Thr, Hyp or an alpha methyl amino acid ( e.g . Aib); and Xaa 37 is Tyr, Pro, Hyp or an alpha methyl amino acid ( e.g. Aib); wherein the polypeptide comprises at least one albumin binding moiety, and wherein the at least one albumin binding moiety comprises a lipid; and wherein the polypeptide comprises any one of the following amino acid modifications or combinations of amino acid modifications: -1G, -2G, 17Aib; 4A, 15W, 21P, 24P, 25A, 28S; 4I, 20I, 21A, 35R; 4I, 21Dab, 35R; 14Dab; 14Dab, 17Aib, 31E; 14Dab, 23αMePhe, 31E; 14Dab, 31E; 14E, 17Aib; 14E, 17Aib, 21H; 14E, 17R; 14E, 17R, 23αMePhe; 14E, 21Aib; 14H, 17Aib; 14H, 17Aib, 21Aib, 31E; 14H, 21Aib; 14H, 21Aib, 35E; 16dL, 21Aib; 17Aib; 17Aib, 37P; 17Aib, 21Aib; 17Aib, 21Aib, 37P; 17Aib, 21G; 17Aib, 21H; 17Aib, 21K; 17Aib, 21P; 17Aib, 21P, 31E; 17Aib, 21P, 35E; 17Aib, 21R; 17Aib, 215; 17Aib, 21Dab; 17Aib, 21Dab, 31E; 17Aib, 22H; 17Aib, 22H, 35E; 17Aib, 23αMePhe; 17Aib, 26Aib; 17Aib, 26R; 17Aib, 27Aib; 17Aib, 27dL; 17Aib, 28Aib; 17Aib, 29Aib; 17Aib, 31Aib; 17Aib, 31E; 17Aib, 31H, 35E; 17Aib, 31P; 17Aib, 31R; 17Aib, 32Aib; 17Aib, 33Aib; 17Aib, 34Aib; 17Aib, 34H; 17Aib, 34P; 17Aib, 35Aib; 17Aib, 35E; 17Aib, 35R; 17E, 21Aib; 17R, 21Aib; 17R, 21Aib, 31Aib; 17R, 21Aib, 31E; 17R, 21Aib, 31R; 17R, 21Aib, 35Aib; 17R, 23αMePhe; 17R, 23αMePhe, 31E; 17R, 26Aib; 17S, 21Aib; 17S, 21Aib, 31H; 17S, 21Aib, 31P; 17S, 21Aib, 31R; 17S, 21Aib, 33P; 17S, 21Aib, 35P; 20αMeSer; 20P, 21P, 24P, 25A, 285; 21Aib; 21Aib, 24P, 25A, 285, 31Dab; 21Aib, 24P, 25A, 285, 35Dab; 21Aib, 26dl; 21Aib, 26Aib; 21Aib, 27Aib; 21Aib, 27dL; 21Aib, 28Aib; 21Aib, 28dP; 21Aib, 31Aib; 21Aib, 31E; 21Aib, 31H; 21Aib, 31R; 21Aib, 33Aib; 21Aib, 34Aib; 21Aib, 35Aib; 21Aib, 35E; 21Aib, 35R; 21Aib, 36Aib; 21Aib, 37Aib; 21Aib, 37P; 21Aib, 24Hyp, 25A, 285; 21Dab, 25Aib; 22Aib; 22H, 35E; 23αMePhe, 31E; 23αMePhe, 31R; 23αMePhe, 35R; 24Aib; 26Aib; 27Aib; Δ1K, 4I, 21Dab, 35R; or Δ1K, 14E, 17R, 23αMePhe.
  2. A polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: wherein: Xaa (-4) is Lys(albumin binding moiety) or is absent; Xaa (-3) is Gly or is absent; Xaa (-2) is Gly or is absent; Xaa (-1) is (albumin binding moiety), Lys(albumin binding moiety) or is absent; Xaa 1 is Lys, Lys(albumin binding moiety), (albumin binding moiety) or is absent; Xaa 4 is Thr; Xaa 14 is Asn, Glu or Dab; Xaa 15 is Phe; Xaa 16 is Leu; Xaa 17 is Val, Arg or Aib; Xaa 20 is Ser; Xaa 21 is Asn; Xaa 22 is Asn; Xaa 23 is Phe or αMePhe; Xaa 24 is Gly; Xaa 25 is Pro; Xaa 26 is Ile; Xaa 27 is Leu; Xaa 28 is Pro; Xaa 29 is Pro; Xaa 31 is Asn, Glu or Arg; Xaa 32 is Val; Xaa 33 is Gly; Xaa 34 is Ser; Xaa 35 is Asn or Arg; Xaa 36 is Thr; and Xaa 37 is Tyr; wherein the polypeptide comprises at least one albumin binding moiety, and wherein the at least one albumin binding moiety comprises a lipid; optionally wherein: (i) Xaa 23 is αMePhe; (ii) Xaa 14 is Glu or Dab; (iii) Xaa 17 is Arg or Aib; (iv) Xaa 31 is Glu or Arg; (v) Xaa 35 is Arg; and/or (vi) Xaa(1) is absent.
  3. The polypeptide or pharmaceutically acceptable salt of any one of the preceding claims, wherein the lipid is attached to the N-terminus of the polypeptide, e.g. wherein the lipid is attached to a lysine at the N-terminus of the polypeptide; optionally wherein: (i) the lipid is attached to the amino acid residue at Xaa -4, Xaa -3, Xaa -2, Xaa -1 or Xaa 1; (ii) the lipid is attached to Xaa -4, Xaa -1 or Xaa 1; optionally wherein the lipid is attached to the N-terminus or to the side-chain of Xaa -4, Xaa -1 or Xaa 1; or (iii) Xaa (-4) is Lys(albumin binding moiety), Xaa (-1) is Lys(albumin binding moiety) or (albumin binding moiety), or Xaa 1 is Lys(albumin binding moiety).
  4. The polypeptide or pharmaceutically acceptable salt of any one of the preceding claims, wherein the lipid comprises a hydrocarbon chain having from 10 to 26 C atoms, e.g. from 14 to 24 C atoms, from 16 to 22 C atoms, or comprising 16, 17, 18, 19, 20, 21, 22, 23 or 24 C atoms; optionally wherein the lipid comprises a dicarboxylic acid; further optionally wherein the lipid is selected from C12diacid, C14diacid, C16diacid, C17diacid, C18diacid, C19diacid or C20diacid.
  5. The polypeptide or pharmaceutically acceptable salt of any one of the preceding claims, wherein the albumin binding moiety is attached to an amino acid residue by a linker; optionally wherein the linker comprises one or more residues of any naturally occurring or non-naturally occurring amino acid; further optionally wherein: (i) the linker comprises a combination of residues, as single or repeating units, each of which may independently be a residue selected from: Glu, γ-Glu, Lys, ε-Lys, Asp, β-Asp, Gaba, β-Ala (3-aminopropanoyl), O2Oc (2-(2-(2-aminoethoxy)ethoxy)acetic acid), PEG2 (3-(2-(2-aminoethoxy)ethoxy)propanoic acid), PEG4 (1-amino-3, 6, 9, 12-tetraoxapentadecan-15-oic acid), PEG8 (1-amino-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-oic acid), or PEG12 (1-amino-3,6,9,12,15,18,21,24,27,30,33,36-dodecaoxanonatriacontan-39-oic acid); or (ii) the linker comprises a residue of γ-Glu, even further optionally wherein the linker comprises yGlu, yGlu-yGlu, γGlu-(O2Oc)-(O2Oc) or yGlu-(PEG2)-(PEG2).
  6. The polypeptide or pharmaceutically acceptable salt of any one of the preceding claims, wherein the polypeptide or pharmaceutically acceptable salt comprises: K(γE-C18diacid)K[CNTATC]ATQRLANFLRHSSNN(αMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 15); K(γE-C18diacid)[CNTATC]ATQRLANFLRHSSNN(αMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 16); K(O2Oc-o2oc-γE-C18diacid)[CNTATC]ATQRLANFLRHSSNN(αMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 17); or K(γE-γE-C18diacid)[CNTATC]ATQRLANFLRHSSNN(αMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 18).
  7. A pharmaceutical composition comprising the polypeptide or pharmaceutically acceptable salt of any one of the preceding claims and a pharmaceutically acceptable excipient.
  8. The polypeptide or pharmaceutically acceptable salt of any one of claims 1-6 or the pharmaceutical composition of claim 7 for use in a method of treating and/or preventing a disease or disorder in a subject, the method comprising administering the polypeptide, pharmaceutically acceptable salt or pharmaceutical composition to a subject in need thereof.
  9. The polypeptide or pharmaceutically acceptable salt for use according to claim 8, or the pharmaceutical composition for use according to claim 8, wherein the disease or disorder is obesity, metabolic disease, an obesity-related condition, eating disorder, Alzheimer's disease, hepatic steatosis ("fatty liver"), kidney failure, arteriosclerosis (e.g. atherosclerosis), cardiovascular disease, macrovascular disease, microvascular disease, diabetic heart (including diabetic cardiomyopathy and heart failure as a diabetic complication), coronary heart disease, peripheral artery disease or stroke, cancer, dumping syndrome, hypertension e.g. pulmonary hypertension, or dyslipidemia e.g. atherogenic dyslipidemia, cholescystitis, or short bowel syndrome.
  10. The polypeptide or pharmaceutically acceptable salt for use according to claim 9, or the pharmaceutical composition for use according to claim 9, wherein: (i) the obesity-related condition is overweight, morbid obesity, obesity prior to surgery, obesity-linked inflammation, obesity-linked gallbladder disease, sleep apnea and respiratory problems, hyperlipidemia, degeneration of cartilage, osteoarthritis, or reproductive health complications of obesity or overweight such as infertility; or (ii) the metabolic disease is diabetes, type 1 diabetes, type 2 diabetes, gestational diabetes, pre-diabetes, insulin resistance, impaired glucose tolerance (IGI), disease states associated with elevated blood glucose levels, metabolic disease including metabolic syndrome, or hyperglycemia e.g. abnormal postprandial hyperglycemia.
  11. The polypeptide or pharmaceutically acceptable salt for use according to any one of claims 8-10, or the pharmaceutical composition for use according to any one of claims 8-10, wherein the polypeptide, pharmaceutically acceptable salt or pharmaceutical composition is administered to the subject by subcutaneous injection.
  12. The polypeptide or pharmaceutically acceptable salt for use according to any one of claims 8-10, or the pharmaceutical composition for use according to any one of claims 8-10, wherein the polypeptide, pharmaceutically acceptable salt or pharmaceutical composition is administered to the subject by self-administration.
  13. A method for the production of the polypeptide of any one of claims 1 to 6; optionally comprising synthesizing the polypeptide by solid-phase or liquid-phase methodology, and optionally isolating and purifying the final product.
  14. An article of manufacture comprising the polypeptide or pharmaceutically acceptable salt of any one of claims 1 to 6 or the pharmaceutical composition of claim 7.
  15. A kit comprising the polypeptide or pharmaceutically acceptable salt of any one of claims 1 to 6 or the pharmaceutical composition of claim 7, optionally further comprising instructions for use.

Description

Field of the Invention The present invention relates to polypeptides which are pramlintide analogues and uses thereof. In particular, the present invention relates to polypeptides which are pramlintide analogues conjugated to half-life extending moieties such as albumin binding moieties and uses thereof. Background Pramlintide is a synthetic analogue of human amylin with three proline substitutions, at positions 25, 28 and 29. As a result of these substitutions, pramlintide has a reduced propensity to form amyloid fibrils, thereby overcoming a physicochemical liability of native human amylin (Kruger DF, Gloster MA. Pramlintide for the treatment of insulin-requiring diabetes mellitus: rationale and review of clinical data. Drugs. 2004; 64(13):1419-32). Pramlintide is clinically used in amylin replacement therapies and simulates the important glucoregulatory actions of amylin. These glucoregulatory actions complement those of insulin by regulating the rate of appearance of glucose in the circulation, and are achieved through three primary mechanisms: slowing the rate of gastric emptying, suppression of post-meal glucagon secretion and suppression of food intake (Roth JD et. al. GLP-1R and amylin agonism in metabolic disease: complementary mechanisms and future opportunities. Br J Pharmacol. 2012;166(1):121-136). Pramlintide has been used as an adjunct to insulin in patients with diabetes who have failed to reach desired glucose control despite optimal insulin therapy (Pullman J, et. al. Pramlintide is used in the management of insulin-using patients with type 2 and type 1 diabetes. Vasc Health Risk Manag. 2006;2(3):203-212). Pharmacokinetic studies show that the terminal half-life of amylin in rats is around 13 minutes, and the half-life for pramlintide in human is ~20-45 minutes (Roth JD et. al. GLP-1R and amylin agonism in metabolic disease: complementary mechanisms and future opportunities. Br J Pharmacol. 2012;166(1):121-136). There remains a need for pramlintide analogues which retain amylin agonist activity and provide advantages such as extended half-life and reduced fibrillation tendency. Summary of Invention The present invention relates to polypeptides that are pramlintide analogues conjugated to albumin binding moieties (e.g. lipids). Thus, in one aspect, there is provided a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: wherein: Xaa (-4) is Lys(albumin binding moiety) or is absent;Xaa (-3) is Gly or is absent;Xaa (-2) is Gly or is absent;Xaa (-1) is Gly, (albumin binding moiety), Lys(albumin binding moiety) or is absent;Xaa 1 is Lys, Lys(albumin binding moiety), (albumin binding moiety) or is absent;Xaa 4 is Thr, Ile or Ala;Xaa 14 is Asn, His, Glu, 2,4-diaminobutanoic acid (Dab), or an alpha methyl amino acid;Xaa 15 is Phe or Trp;Xaa 16 is Leu or D-Leu (dL);Xaa 17 is Val, Ser, Glu, Arg, (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid (Hyp), Dab or an alpha methyl amino acid (e.g. 2-amino-2-methylpropanoic acid [Aib]);Xaa 20 is Ser, Ile, Pro or an alpha methyl amino acid (e.g. (S)-2-amino-3-hydroxy-2-methylpropanoic acid [aMeSer]);Xaa 21 is Asn, Dab, His, Pro, Ser, Arg, Lys, Gly, Glu, Ala, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 22 is Asn, His, Hyp, Dab or an alpha methyl amino acid (e.g. Aib);Xaa 23 is Phe, Hyp or an alpha methyl amino acid (e.g. (S)-2-amino-2-methyl-3-phenylpropanoic acid [αMePhe]);Xaa 24 is Gly, Pro, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 25 is Pro, Ala, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 26 is Ile, D-Ile (dl), Arg, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 27 is Leu, dL, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 28 is Pro, D-Pro (dP), Ser, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 29 is Pro, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 31 is Asn, Glu, His, Arg, Pro, Dab or an alpha methyl amino acid (e.g. Aib);Xaa 32 is Val, Hyp, Dab or an alpha methyl amino acid (e.g. Aib);Xaa 33 is Gly, Pro, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 34 is Ser, Pro, His, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 35 is Asn, Pro, Arg, Glu, Dab, Hyp or an alpha methyl amino acid (e.g. Aib);Xaa 36 is Thr, Hyp or an alpha methyl amino acid (e.g. Aib); andXaa 37 is Tyr, Pro, Hyp or an alpha methyl amino acid (e.g. Aib), and wherein the polypeptide comprises at least one albumin binding moiety. In another aspect, there is provided a lipidated polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence: wherein: Xaa (-4) is Lys(linker-lipid) or is absent;Xaa (-3) is Gly or is absent;Xaa (-2) is Gly or is absent;Xaa (-1) is Gly, (linker-lipid), Lys(linker-lipid) or is absent;Xaa 1 is Lys, Lys(linker-lipid), (linker-lipid) or is absent;Xaa 4 is Thr, Ile or Ala;Xaa 14 is Asn, His, Glu, 2,4-diaminobutanoic acid (Dab), or an alpha methyl amino acid;Xaa 15 is Phe or Trp;Xaa 16 is Leu or D-Leu (dL);Xaa 17 is Val, Ser, Glu, Arg, (2S,4R)-4-hydroxypyrrolidi