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WO2010013012A4 - Hypothermia inducing polypeptides and uses thereof - Google Patents

Hypothermia inducing polypeptides and uses thereof Download PDF

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Publication number
WO2010013012A4
WO2010013012A4 PCT/GB2009/001885 GB2009001885W WO2010013012A4 WO 2010013012 A4 WO2010013012 A4 WO 2010013012A4 GB 2009001885 W GB2009001885 W GB 2009001885W WO 2010013012 A4 WO2010013012 A4 WO 2010013012A4
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WO
WIPO (PCT)
Prior art keywords
polypeptide
seq
polypeptide according
amino acid
subject
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB2009/001885
Other languages
French (fr)
Other versions
WO2010013012A9 (en
WO2010013012A3 (en
WO2010013012A2 (en
Inventor
Carin SJÖLUND
Björn Ulrik WALSE
Tadeusz Wieloch
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
LUND UNIVERSITY BIOSCIENCE AB
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LUND UNIVERSITY BIOSCIENCE AB
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Filing date
Publication date
Application filed by LUND UNIVERSITY BIOSCIENCE AB filed Critical LUND UNIVERSITY BIOSCIENCE AB
Publication of WO2010013012A2 publication Critical patent/WO2010013012A2/en
Publication of WO2010013012A3 publication Critical patent/WO2010013012A3/en
Publication of WO2010013012A4 publication Critical patent/WO2010013012A4/en
Publication of WO2010013012A9 publication Critical patent/WO2010013012A9/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575Hormones
    • C07K14/57563Vasoactive intestinal peptide [VIP]; Related peptides
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575Hormones
    • C07K14/605Glucagons
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Endocrinology (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Vascular Medicine (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The present invention provides a polypeptide capable of binding to a receptor for glucagon-like peptide-1 (GLP-1) for use in inducing or maintaining hypothermia in a subject in need thereof, wherein the polypeptide comprises or consists of an amino acid sequence of SEQ ID NO: 1 or a fragment, variant, derivative or fusion thereof (or a fusion of said fragment, variant or derivative) which retains the hypothermia-inducing activity of said amino acid sequence. The invention further provides pharmaceutical compositions comprising the polypeptides of the invention, together with methods and uses of the same.

Claims

AMENDED CLAIMS received by the International Bureau on 12 april 2010 (12.04.10)
1. A polypeptide capable of binding to a receptor for glucagon-like peptide-1 (GLP-1) for use in inducing or maintaining hypothermia in a subject in need thereof,
wherein the polypeptide comprises or consists of an amino acid sequence of SEQ ID NO: 1 :
AU-01: HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS [SEQ ID
NO: 1]
or a fragment, variant, derivative or fusion thereof (or a fusion of said fragment, variant or derivative) which retains the hypothermia-inducing activity of said amino acid sequence.
2. A polypeptide according to Claim 1 for use in the treatment or prevention of neuronal damage in the central nervous system.
3. A polypeptide according to Claim 1 or 2 for use in the treatment or prevention of acute brain injury.
4. A polypeptide according to any one of the preceding claims for use in the treatment or prevention of neuronal damage due to ischemia.
5. A polypeptide according to any one of the preceding claims wherein the subject is suffering from or has recently suffered from a stroke, a brain trauma, a cardiac arrest, spinal cord injury or asphyxia.
6. A polypeptide according to any one of the preceding claims wherein the subject is suffering from or has recently suffered from a cardiac arrest.
7. A polypeptide according to Claim 5 or 6 wherein the subject is suffering from or has recently suffered from a myocardial infarction.
8. A polypeptide according to any one of the preceding claims wherein the polypeptide is for use in combination with one or more additional hypothermic treatments.
66
. A polypeptide according to Claim 8 wherein the one or more additional hypothermic treatments include external and/or internal surface cooling.
10. A polypeptide according to Claim 8 or 9 wherein the one or more additional hypothermic treatments include nasopharyngeal cooling and/or cold saline infusion.
11. A polypeptide according to Claim 8 or 9 wherein the one or more additional hypothermic treatments is selected from the group consisting of vanilloid receptor agonists, cannabinoid receptor agonists adenosine receptor agonists and fever reducing agents such as regulators of prostaglandin synthesis.
12. A polypeptide according to any one of the preceding claims wherein the subject is human.
13. A polypeptide according to any one of Claims 1 to 11 wherein the subject is non- human.
14. A polypeptide according to any one of the preceding claims wherein the polypeptide is capable of inducing a fall of at least 1 °C in core body temperature of the subject, for example a fall of at least 20C, 2.5°C, 3°C, 4°C, 5°C or more.
15. A polypeptide according to any one of the preceding claims wherein the polypeptide is capable of inducing a hypothermic effect having a duration of at least 10 minutes, for example at least 20 minutes, 30minutes, 40 minutes, 50 minutes, 60 minutes, 90 minutes, 120 minutes or more.
16. A polypeptide according to any one of the preceding claims wherein the polypeptide has an in vivo half-life in humans of at least 10 minutes following IV administration, for example at least 15 minutes or at least 20 minutes or more.
17. A polypeptide according to any one of the preceding claims wherein the polypeptide has comparable efficacy in healthy subjects and in subjects suffering from neuronal damage due to an ischemia.
18. A polypeptide according to any one of the preceding claims wherein the polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 1.
67
19. A polypeptide according to any one of Claims 1 to 17 wherein the polypeptide comprises or consists of a fragment of the amino acid sequence according to SEQ ID NO: 1.
20. A polypeptide according to Claim 19 wherein the fragment comprises or consists of at least 10 contiguous amino acid of SEQ ID NO: 1 , for example at least 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37 or 38 contiguous amino acid of SEQ ID NO: 1.
21. A polypeptide according to Claim 19 or 20 wherein the fragment commences at an amino acid residue selected from amino acid residues 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 ,
12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 and 29 of SEQ ID NO:1.
22. A polypeptide according to any one of Claims 19 to 21 wherein the fragment terminates at an amino acid residue selected from amino acid residues 10, 11 , 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38 and 39 of SEQ ID NO:1.
23. A polypeptide according to any one of Claims 19 to 22 wherein the fragment comprises or consists of amino acids 3 to 8 and/or 15 to 32 of SEQ ID NO: 1.
24. A polypeptide according to any one of Claims 1 to 17 wherein the polypeptide comprises or consists of a variant of the amino acid sequence according to SEQ ID NO: 1.
25. A polypeptide according to Claim 24 wherein the variant is a non-naturally occurring variant.
26. A polypeptide according to Claim 24 or 25 wherein the variant has an amino acid sequence which has at least 50% identity with the amino acid sequence according to SEQ ID NO: 1 or a fragment thereof, for example at least 55%, 60%, 65%, 70%, 75%, 80%, 90%, 95%, 96%, 97%, 98% or at least 99% identity.
27. A polypeptide according to any one of Claims 24 to 26 wherein the variant comprises or consists of an amino acid sequence of SEQ ID NO: 2:
68 His-XaaZ-XaaS-Gly-Thr-Phe-Thr-Ser-Asp-XaalO-Ser-Xaa^-XaalS-XaaM-Glu-Glu- Glu-Ala-Val~Arg-Leu-Phe-lle-Glu-Trp-Leu-Lys- Xaa28-G)y-Gly-Pro-Ser-Ser-Gly-Ala~ Xaa36-Pro-Pro-Ser-Xaa40-Xaa41-Xaa42-Xaa43-Xaa44-Xaa45-Xaa46
[SEQ ID N0:2]
wherein
Xaa2 is GIy, Ser, VaI, Ala or aminoisobutyric acid (Aib);
Xaa3 is GIu or Asp;
Xaa10 is Leu or VaI;
Xaa12 is Lys or Ser;
Xaa13 is Gln, Glu or Tyr;
Xaa14 is Met or Leu
Xaa28 is Asn, Ser or Asp;
Xaa36 is Pro or absent;
Xaa40 is an amide group, Lys, AEEA-MPA-modified Lys or is absent;
Xaa41 is Lys or absent;
Xaa42 is Lys or absent;
Xaa43 is Lys or absent;
Xaa44 is Lys or absent;
Xaa45 is Lys or absent; and
Xaa46 is an amide group or is absent
or a fragment, derivative or fusion thereof (or a fusion of said fragment or derivative) which retains the hypothermia-inducing activity of a polypeptide having the amino acid sequence of SEQ ID NO:1.
28. A polypeptide according to Claim 27 wherein the variant comprises or consists of an amino acid sequence of SEQ ID NO: 2.
29. A polypeptide according to Claim 27 or 28 wherein the variant comprises or consists of an amino acid sequence selected from the group consisting of:
AU-08: HSDGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:4],
AU-11: HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGA-PPSKKKKKK
[SEQ ID NO:5];
69 AU-12: HGEGTFTSDLSKEMEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:6];
AU-U: HGEGTFTSDLSKQMEEEAVRLFIEWLKDGGPSSGAPPPS
[SEQ ID NO:7];
AU- 15: HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPSZ, where Z=Lys-(AEEA-MPA)-(CONH2) [SEQ ID NO:8];
AU-M: HAEGTFTSDVSSYLEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:9],
AU-18: HGEGTFTSDVSSYLEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:10];
AU-19: HVEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:11];
AU-20: H/\ώEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:12]; and
AU-28: HGEGTFTSDLSKQLEEEAVRLFIEWLKSGGPSSGAPPPS
[SEQ ID NO:13].
30. A polypeptide according to any one of the preceding claims wherein the polypeptide comprises or consists of a fusion protein.
31. A polypeptide according to Claim 30 comprising or consisting of an amino acid sequence corresponding to a fragment of SEQ ID NO: 1 and an amino acid sequence corresponding to a fragment of GLP-1.
32. A polypeptide according to Claim 31 comprising or consisting of an amino acid sequence of SEQ ID NO: 9:
AU-18: HGEGTFTSDVSSYLEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:10].
33. A polypeptide according to any one of the preceding claims wherein the polypeptide, or fragment, variant, fusion or derivative thereof, comprises or consists of L-amino acids.
34. A polypeptide according to any one of the preceding claims wherein the polypeptide, or fragment, variant, fusion or derivative thereof, comprises one or more amino acids that are modified or derivatised.
70
35. A polypeptide according to Claim 34 wherein the one or more amino acids are modified or derivatised by PEGylation, amidation, esterification, acylation, acetylation and/or alkylation.
36. A polypeptide according to any one of the preceding claims wherein the fragment, variant, fusion or derivative exhibits a functional advantage relative to a polypeptide of SEQ ID NO:1.
37. A polypeptide according to Claim 36 wherein the functional advantage is selected from the group consisting of:
(a) Increased receptor affinity for GLP1 receptors;
(b) Reduced vulnerability to DPP-IV degradation; and
(c) Decreased rate of renal clearance.
38. A polypeptide according to any one of the preceding claims wherein the polypeptide is between 10 and 100 amino acids in length, for example between 20 and 60, 30 and 50, 35 and 45, or 38 and 40 amino acids in length.
39. A polypeptide according to Claim 38 wherein the polypeptide is 39 amino acids in length.
40. A polypeptide according to any one of the preceding claims wherein the polypeptide is linear.
41. A polypeptide according to any one of the preceding claims wherein the polypeptide is a recombinant polypeptide.
42. A polypeptide according to any one of the preceding claims wherein the polypeptide comprises an amide group at its C-terminus.
43. An isolated polypeptide comprising or consisting of an amino acid sequence according to SEQ ID NO:2, with the proviso that the isolated polypeptide does not consist of an amino acid sequence according to SEQ ID NO:1.
44. A polypeptide according to Claim 43 wherein the polypeptide is selected from the group consisting of:
71 AU-18: HGEGTFTSDVSSYLEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:10],
AU-19: HVEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:11];and
AU-20: HΛ/όEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS
[SEQ ID NO:12]; and
AU-28: HGEGTFTSDLSKQLEEEAVRLFIEWLKSGGPSSGAPPPS
[SEQ ID NO:13].
45. A polypeptide according to Claim 43 or 44 wherein the polypeptide comprises an amide group at its C-terminus.
46. An isolated nucleic acid molecule encoding a polypeptide according to any one of Claims 43 to 45.
47. A vector comprising a nucleic acid molecule according to Claim 46.
48. A vector according to Claim 47 wherein the vector is an expression vector.
49. A host cell comprising a vector according to Claim 47 or 48.
50. A pharmaceutical composition comprising a polypeptide as defined in any one of Claims 1 to 45 together with a pharmaceutically acceptable excipient, diluent or carrier.
51. A pharmaceutical composition according to Claim 50 suitable for parenteral administration.
52. A pharmaceutical composition according to Claim 51 suitable for intravenous, intracerebroventricular, intratechal and/or sub-cutaneous administration.
53. Use of a polypeptide according to any one of Claims 1 to 45 in the preparation of a medicament inducing or maintaining hypothermia in a subject in need thereof.
54. A use according to Claim 53 wherein the medicament is for the treatment or prevention of neuronal damage in the central nervous system.
55. A use according to Claim 53 or 54 wherein the medicament is for use in the treatment or prevention of acute brain injury.
56. A use according to any one of Claims 53 to 55 wherein the medicament is for use in the treatment or prevention of neuronal damage due to ischemia.
57. A use according to any one of Claims 53 to 56 wherein the subject is suffering from or has recently suffered from a stroke, a brain trauma, a cardiac arrest, spinal cord injury or asphyxia.
58. A use according to any one of Claims 53 to 57 wherein the subject is suffering from or has recently suffered from a cardiac arrest.
59. A use according to Claim 57 or 58 wherein the subject is suffering from or has recently suffered from a myocardial infarction.
60. A method for inducing or maintaining hypothermia in a subject in need thereof, the method comprising administering to the patient a therapeutically-effective amount of a polypeptide according to any one of Claims 1 to 45.
61. A method according to Claim 60 wherein the subject is human.
62. A method according to Claim 60 or 61 wherein the subject is conscious during administration of the polypeptide.
63. A method according to any one of Claims 60 to 62 wherein the medicament is for the treatment or prevention of neuronal damage in the central nervous system.
64. A method according to any one of Claims 60 to 63 wherein the medicament is for use in the treatment or prevention of acute brain injury.
65. A method according to any one of Claims 60 to 64 wherein the medicament is for use in the treatment or prevention of neuronal damage due to ischemia.
73
66. A method according to any one of Claims 60 to 65 wherein the subject is suffering from or has recently suffered from a stroke, a brain trauma, a cardiac arrest, spinal cord injury or asphyxia.
67. A method according to any one of Claims 60 to 66 wherein the subject is suffering from or has recently suffered from a cardiac arrest.
68. A method according to Claim 66 or 67 wherein the subject is suffering from or has recently suffered from a myocardial infarction.
69. Method of inducing and/or maintaining hypothermia in a patient, such as a mammal, exposed to brain injury, comprising administration of a therapeutically effective amount to the patient of a composition having a component that binds to a receptor for glucagon-like peptide-1 (GLP-1).
70. The method according to claim 69, further comprising exposing said mammal to other hypothermic treatment, such as external surface cooling or internal surface cooling.
71. The method according to claim 70, wherein said hypothermic treatment comprises nasopharyngeal cooling and/or cold saline infusion.
72. The method according to claim 70 or 71, wherein said administration and said other hypothermic treatment takes place simultaneously.
73. The method according to any one of Claims 69 to 72, wherein said composition comprises a substance included in the group comprising:
exendin-4; a human glucagon-like peptide (GLP)-I -albumin recombinant protein
(Albugon);
CJC-1131; liraglutide; and oxyntomodulin.
74
74. Use of a substance as defined in Claim 73, for the preparation of a medicament for the induction and/or maintenance of hypothermia for treatment of brain injury, particularly stroke, brain trauma, cardiac arrest and asphyxia.
75. The use according to claim 74, wherein the use is combined with hypothermic treatment according to claim 71 or 72.
76. A polypeptide substantially as described herein with reference to the description and figures.
77. A nucleic acid molecule as described herein with reference to the description and figures.
78. A vector as described herein with reference to the description and figures.
79. A host cell as described herein with reference to the description and figures.
80. A pharmaceutical composition substantially as described herein with reference to the description and figures.
81. Use of a polypeptide substantially as described herein with reference to the description and figures.
82. A method for inducing or maintaining hypothermia as described herein with reference to the description and figures.
83. A method for diagnosis substantially as described herein with reference to the description and figures.
75
PCT/GB2009/001885 2008-08-01 2009-07-31 Novel polypeptides and uses thereof Ceased WO2010013012A2 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
SE0801752-7 2008-08-01
SE0801752 2008-08-01
US19179108P 2008-09-12 2008-09-12
US61/191,791 2008-09-12

Publications (4)

Publication Number Publication Date
WO2010013012A2 WO2010013012A2 (en) 2010-02-04
WO2010013012A3 WO2010013012A3 (en) 2010-04-01
WO2010013012A4 true WO2010013012A4 (en) 2010-06-10
WO2010013012A9 WO2010013012A9 (en) 2010-08-12

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WO2010120476A2 (en) * 2009-04-01 2010-10-21 Amylin Pharmaceuticals, Inc. N-terminus conformationally constrained glp-1 receptor agonist compounds
EP2718318B1 (en) 2011-06-10 2018-07-25 Hanmi Science Co., Ltd. Novel oxyntomodulin derivatives and pharmaceutical composition for treating obesity comprising the same
UA114709C2 (en) 2011-06-17 2017-07-25 Ханмі Сайенс Ко., Лтд. CONJUGATE CONTAINING OXYNTOMODULIN DERIVATIVES, THE FACILITY OF THE IMMUNOGLOBULIN AND THE NON-REPTIDYL POLYMER, AND ITS APPLICATION
KR101968344B1 (en) 2012-07-25 2019-04-12 한미약품 주식회사 A composition for treating hyperlipidemia comprising oxyntomodulin analog
UA116217C2 (en) 2012-10-09 2018-02-26 Санофі Exendin-4 derivatives as dual glp1/glucagon agonists
KR101993393B1 (en) 2012-11-06 2019-10-01 한미약품 주식회사 A composition for treating diabetes or diabesity comprising oxyntomodulin analog
SG10201602801YA (en) 2012-11-06 2016-05-30 Hanmi Pharm Ind Co Ltd Liquid formulation of protein conjugate comprising the oxyntomodulin and an immunoglobulin fragment
SI2934568T1 (en) 2012-12-21 2018-03-30 Sanofi Dual glp1/gip or trigonal glp1/gip/glucagon agonists
TW201609797A (en) 2013-12-13 2016-03-16 賽諾菲公司 Dual GLP-1/glucagon receptor agonists
TW201609796A (en) 2013-12-13 2016-03-16 賽諾菲公司 Non-acylated EXENDIN-4 peptide analogues
TW201609795A (en) 2013-12-13 2016-03-16 賽諾菲公司 EXENDIN-4 peptide analogues as dual GLP-1/GIP receptor agonists
WO2015086729A1 (en) 2013-12-13 2015-06-18 Sanofi Dual glp-1/gip receptor agonists
TW201625668A (en) 2014-04-07 2016-07-16 賽諾菲公司 Exendin-4 derivatives as peptidic dual GLP-1/glucagon receptor agonists
TW201625670A (en) 2014-04-07 2016-07-16 賽諾菲公司 Dual GLP-1/glucagon receptor agonists derived from EXENDIN-4
TW201625669A (en) 2014-04-07 2016-07-16 賽諾菲公司 Peptidic dual GLP-1/glucagon receptor agonists derived from Exendin-4
US9932381B2 (en) 2014-06-18 2018-04-03 Sanofi Exendin-4 derivatives as selective glucagon receptor agonists
TWI802396B (en) 2014-09-16 2023-05-11 南韓商韓美藥品股份有限公司 Use of a long acting glp-1/glucagon receptor dual agonist for the treatment of non-alcoholic fatty liver disease
KR102418477B1 (en) 2014-12-30 2022-07-08 한미약품 주식회사 Gluagon Derivatives
KR101661332B1 (en) * 2015-05-28 2016-09-29 (의료)길의료재단 Pharmaceutical composition for treating muscle atrophy comprising glucagon like-peptide-1 receptor agonist
AR105319A1 (en) 2015-06-05 2017-09-27 Sanofi Sa PROPHARMS THAT INCLUDE A DUAL AGONIST GLU-1 / GLUCAGON CONJUGATE HIALURONIC ACID CONNECTOR
TW201706291A (en) 2015-07-10 2017-02-16 賽諾菲公司 New EXENDIN-4 derivatives as selective peptidic dual GLP-1/glucagon receptor agonists

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WO2010013012A3 (en) 2010-04-01
WO2010013012A2 (en) 2010-02-04

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