US2014105991A1PendingUtilityA1
Spherical microcapsules comprising glp-1 peptides, their production and use
Est. expiryMay 10, 2026(expired)· nominal 20-yr term from priority
A61P 3/04A61P 7/04A61P 5/50A61P 9/14A61P 43/00A61P 3/08A61P 9/10A61P 3/10A61P 25/00A61P 25/16A61P 25/28A61P 21/02A61P 21/00C12N 5/0653A61K 38/26C12N 2510/02A61K 35/35A61K 9/5036A61K 35/12A61K 35/32C12N 2501/33C12N 2501/39C12N 2510/04A61K 35/30C12N 5/0663A61K 9/5052A61K 2035/128A61K 2035/124A61K 9/0024
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Claims
Abstract
The present invention provides spherical microcapsules comprising at least one surface coating and a core, wherein the at least one surface coating comprises cross-linked polymers, and wherein the core comprises cross-linked polymers and cells capable of expressing and secreting a GLP-1 peptide, a fragment or variant thereof or a fusion peptide comprising GLP-1 or a fragment or variant thereof. The present applicators is furthermore directed to methods for production of these spherical microcapsules and to the use of these microcapules e.g. in the treatment of type 2 diabetes, weight disorders and diseases or conditions associated thereto, neurodegenerative disorders and diseases or conditions associated thereto, or for the treatment of disorders and diseases or conditions associated to apoptosis.
Claims
exact text as granted — not AI-modified1 - 34 . (canceled)
35 . A spherical or non-spherical microcapsule comprising at least one surface coating and a core, wherein the at least one surface coating comprises cross-linked polymers, and wherein the core comprises cross-linked polymers and cells capable of expressing and secreting a fusion peptide, wherein the fusion peptide comprises: components (I) and (II), wherein:
(I) component (I) comprises:
(a) N-terminally a GLP-1 (7-35, 7-36 or 7-37) sequence or a fragment or variant thereof; or
(b) a peptide comprising the sequence according to formula II:
(SEQ ID NO: 28)
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa16-Ser-
Xaa18-Xaa19-Xaa20-Glu-Xaa22-Xaa23-Ala-Xaa25-Xaa26-
Xaa27-Phe-Ile-Xaa30-Trp-Leu-Xaa33-Xaa34-Xaa35-
Xaa36-Xaa37,
wherein
Xaa7 is L-histidine) D-histidine, desamino-histidine, 2-aminohistidine, 3-hydroxy-histidine, homohistidine, N-acetyl-histidine, α-fluoromethyl-histidine, α-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine;
Xaa8 is Ala, Gly, Val, Leu, Ile, Lys, Aib, (1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
Xaa16 is Val or Leu;
Xaa18 is Ser, Lys or Arg;
Xaa19 is Tyr or Gln;
Xaa20 is Leu or Met;
Xaa22 is Gly, Glu, or Aib;
Xaa23 is Gln, Glu, Lys or Arg;
Xaa25 is Ala or Val;
Xaa26 is Lys, Glu or Arg;
Xaa27 is Glu or Leu;
Xaa30 is Ala, Glu or Arg;
Xaa33 is Val or Lys;
Xaa34 is Lys, Glu, Asn or Arg;
Xaa35 is Gly or Aib;
Xaa36 is Arg, Gly or Lys or amide or absent;
Xaa37 is Gly, Ala, Glu, Pro, Lys, amide or is absent; or
(c) a peptide comprising a sequence according to formula III:
(SEQ ID NO: 29)
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-
Xaa18-Tyr-Leu-Glu-Xaa22-Xaa23-Ala-Ala-Xaa26-Glu-
Phe-Ile-Xaa30-Trp-Leu-Val-Xaa34-Xaa35-Xaa36-Xaa37,
wherein
Xaa7 is L-histidine, D-histidine, desamino-histidine, 2-aminohistidine, 3-hydroxy-histidine, homohistidine, N-acetyl-histidine, α-fluoromethyl-histidine, α-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine;
Xaa8 is Ala, Gly, Val, Leu, Ile, Lys, Aib, (1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
Xaa18 is Ser, Lys or Arg;
Xaa22 is Gly, Glu or Aib;
Xaa23 is Gln, Glu, Lys or Arg;
Xaa26 is Lys, Glu or Arg;
Xaa30 is Ala, Glu or Arg;
Xaa34 is Lys, Glu or Arg;
Xaa35 is Gly or Aib;
Xaa36 is Arg or Lys, amide or is absent;
Xaa37 is Gly, Ala, Glu or Lys, amide or is absent, and
(II) component (II) comprises:
C-terminally a peptide sequence of at least 9 amino acids,
wherein component (II) is a peptide sequence containing a sequence according to SEQ ID NO: 22 (RRDFPEEVAI) or a sequence having at least 80% sequence homology with SEQ ID NO: 22.
36 . The spherical or non-spherical microcapsule according to claim 35 , wherein the cross-linked polymer of the core and/or the at least one surface coating comprises biopolymers.
37 . The spherical or non-spherical microcapsule according to claim 35 , wherein the cross-linked polymer of the core and/or the at least one surface coating comprises an alginate.
38 . The spherical or non-spherical microcapsule according to claim 35 , wherein the cross-linked polymer of the core and/or the at least one surface coating comprises a chemically identical polymer in identical or differing concentrations, wherein the polymers further may have different molecular weights and/or may be cross-linked differently.
39 . The spherical or non-spherical microcapsule according to claim 35 , wherein the core has a diameter of about 20 to about 2000 μm and wherein the at least one surface coating has a thickness of about 10 to about 2000 μm.
40 . The spherical or non-spherical microcapsule according to claim 35 , wherein the microcapsule comprises 1, 2, 3, 4, 5, 5-10 or more surface coatings.
41 . The spherical or non-spherical microcapsule according to claim 35 , wherein the microcapsule comprises an additional external surface coating consisting of polycations.
42 . The spherical or non-spherical microcapsule according to claim 41 , wherein the additional external surface coating consists of Poly-L-Lysine,
43 . The spherical or non-spherical microcapsule according to claim 35 , wherein the GLP-1 peptide is selected from the group consisting of a peptide comprising aa 7-35 of GLP-1 and a peptide showing a homology of at least 80% with this peptide.
44 . The spherical or non-spherical microcapsule according to claim 35 , wherein component (I) of the fusion peptide contains a sequence having at least 80% sequence homology with SEQ ID NO: 1.
45 . The spherical or non-spherical microcapsule according to claim 35 , wherein component (II) is a peptide sequence containing a sequence according to SEQ ID NO: 23 (RRDFPEEVAIVEEL) or SEQ ID NO: 24 (RRDFPEEVAIAEEL) or a sequence having at least 80% sequence homology with SEQ ID NOs: 23 or 24.
46 . The spherical or non-spherical microcapsule according to claim 35 , wherein component (II) is a peptide sequence containing a sequence according to SEQ ID NO: 2 (RRDFPEEVAIVEELG) or SEQ ID NO: 3 (RRDFPEEVAIAEELG) or a sequence having at least 80% sequence homology with SEQ ID NOs: 2 or 3.
47 . The spherical or non-spherical microcapsule according to claim 35 , wherein component (I) and component (II) are directly linked or linked via a linker sequence.
48 . The spherical microcapsule according to claim 47 , wherein the linker sequence has a length of 1 to 10 amino acids.
49 . The spherical or non-spherical microcapsule according to claim 35 , wherein the fusion peptide contains a sequence according to SEQ ID NO: 8 (HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIAEELG) or SEQ ID NO: 12 (HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIVEELG) or a sequence having at least 80% sequence homology with SEQ ID NOs: 8 or 12.
50 . The spherical or non-spherical microcapsule according to claim 35 wherein the fusion peptide contains alternatively or additionally to component (II) a component (III), wherein component (III) may be linked to the C-terminus of component (I) and/or to the N-terminus of component (I), if components (I) and (III) are present in the fusion protein, or wherein component (III) may be linked to the C-terminus of component (II) and/or to the N-terminus of component (I), if components (I), (II) and (III) are present in the fusion protein.
51 . The spherical or non-spherical microcapsule according to claim 50 , wherein component (III) comprises at least four amino acid residues to at least 30 amino acid residues.
52 . The spherical or non-spherical microcapsule according to claim 35 , further comprising component (III), wherein component (III) comprises:
(a) at least four to at least 20 additional amino acid residues of the N-terminal sequence of GLP-2 as in proglucagon; or (b) GLP-1(5-37, 6-37, or 7-37); or (c) a peptide comprising the sequence according to formula II:
(SEQ ID NO: 28)
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa16-Ser-
Xaa18-Xaa19-Xaa20-Glu-Xaa22-Xaa23-Ala-Xaa25-Xaa26-
Xaa27-Phe-Ile-Xaa30-Trp-Leu-Xaa33-Xaa34-Xaa35-
Xaa36-Xaa37,
wherein
Xaa7 is L-histidine, D-histidine, desamino-histidine, 2-amino-histidine, 3-hydroxy-histidine, homohistidine, N-acetyl-histidine, α-fluoromethyl-histidine, α-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine;
Xaa8 is Ala, Gly, Val, Leu, He, Lys, Aib, (1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
Xaa16 is Val or Leu;
Xaa18 is Ser, Lys or Arg;
Xaa19 is Tyr or Gln;
Xaa20 is Leu or Met;
Xaa22 is Gly, Glu or Aib;
Xaa23 is Gln, Glu, Lys or Arg;
Xaa25 is Ala or Val;
Xaa26 is Lys, Glu or Arg;
Xaa27 is Glu or Leu;
Xaa30 is Ala, Glu or Arg;
Xaa33 is Val or Lys;
Xaa34 is Lys, Glu, Asn or Arg;
Xaa35 is Gly or Aib;
Xaa36 is Arg, Gly or Lys or amide or absent;
Xaa37 is Gly, Ala, Glu, Pro, Lys, amide or is absent; or
(d) a peptide comprising a sequence according to formula III:
(SEQ ID NO: 29)
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-
Xaa18-Tyr-Leu-Glu-Xaa22-Xaa23-Ala-Ala-Xaa26-Glu-
Phe-Ile-Xaa30-Trp-Leu-Val-Xaa34-Xaa35-Xaa36-Xaa37,
wherein
Xaa7 is L-histidine, D-histidine, desamino-histidine, 2-amino-histidine, 3-hydroxy-histidine, homohistidine, N-acetyl-histidine, α-fluoromethyl-histidine, α-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine;
Xaa8 is Ala, Gly, Val, Leu, Ile, Lys, Aib, (1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
Xaa18 is Ser, Lys or Arg;
Xaa22 is Gly, Glu or Aib;
Xaa23 is Gln, Glu, Lys or Arg;
Xaa26 is Lys, Glu or Arg;
Xaa30 is Ala, Glu or Arg;
Xaa34 is Lys, Glu or Arg;
Xaa35 is Gly or Aib;
Xaa36 is Arg or Lys, amide or is absent; and
Xaa37 is Gly, Ala, Glu or Lys, amide or is absent.
53 . The spherical or non-spherical microcapsule according to claim 50 , wherein component (III) contains the sequence of SEQ ID NOs: 4 or 5 or a sequence having at least 80% sequence homology with SEQ ID NOs: 4 or 5.
54 . The spherical or non-spherical microcapsule according to claim 50 , wherein the fusion peptide contains a peptide sequence selected from a group consisting of: SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 26, or a sequence having at least 80% sequence homology with SEQ ID NOs: 6, 7, 10, 11 or 26.
55 . The spherical or non-spherical microcapsule according to claim 35 wherein the fusion peptide comprises a carrier protein, in particular transferrin or albumin, as component (IV).
56 . The spherical or non-spherical microcapsule according to claim 35 , wherein the cells contained in the core of the spherical microcapsule are selected from human mesenchymal stem cells, differentiated cells derived from human mesenchymal stem cells, including osteoblasts, chondrocytes, fat cells (adipocytes), or neuron-like cells including brain cells.
57 . The spherical or non-spherical microcapsule according to claim 56 , wherein the cells contained in the core of the spherical microcapsule are selected (in vitro) from human mesenchymal stem cells, wherein the cells differentiate in vitro or in vivo into fat cells (adipocytes), suitable for transplantation into fat tissue.
58 . A pharmaceutical composition comprising a spherical or non-spherical microcapsule according to claim 35 and optionally a pharmaceutically acceptable carrier.
59 . A method for producing a spherical or non-spherical microcapsule comprising at least one surface coating and a core, wherein the at least one surface coating comprises a cross-linked polymer, and wherein the core comprises cross-linked polymers and biological cells capable of expressing and secreting a fusion peptide, wherein the fusion peptide comprises: components (I) and (II), wherein:
(I) component ( 1 ) comprises:
(a) N-terminally a GLP-1 (7-35, 7-36 or 7-37) sequence or a fragment or variant thereof; or
(b) a peptide comprising the sequence according to formula II:
(SEQ ID NO: 28)
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa16-Ser-
Xaa18-Xaa19-Xaa20-Glu-Xaa22-Xaa23-Ala-Xaa25-Xaa26-
Xaa27-Phe-Ile-Xaa30-Trp-Leu-Xaa33-Xaa34-Xaa35-
Xaa36-Xaa37,
wherein
Xaa7 is L-histidine, D-histidine, desamino-histidine, 2-aminohistidine, 3-hydroxy-histidine, homohistidine, N-acetyl-histidine, α-fluoromethyl-histidine, α-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine;
Xaa8 is Ala, Gly, Val, Leu, Ile, Lys, Aib, (1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
Xaa16 is Val or Leu;
Xaa18 is Ser, Lys or Arg;
Xaa19 is Tyr or Gln;
Xaa20 is Leu or Met;
Xaa22 is Gly, Glu, or Aib;
Xaa23 is Gln, Glu, Lys or Arg;
Xaa25 is Ala or Val;
Xaa26 is Lys, Glu or Arg;
Xaa27 is Glu or Leu;
Xaa30 is Ala, Glu or Arg;
Xaa33 is Val or Lys;
Xaa34 is Lys, Glu, Asn or Arg;
Xaa35 is Gly or Aib;
Xaa36 is Arg, Gly or Lys or amide or absent;
Xaa37 is Gly, Ala, Glu, Pro, Lys, amide or is absent; or
(c) a peptide comprising a sequence according to formula III:
(SEQ ID NO: 29)
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-
Xaa18-Tyr-Leu-Glu-Xaa22-Xaa23-Ala-Ala-Xaa26-Glu-
Phe-Ile-Xaa30-Trp-Leu-Val-Xaa34-Xaa35-Xaa36-Xaa37,
wherein
Xaa7 is L-histidine, D-histidine, desamino-histidine, 2-aminohistidine, 3-hydroxy-histidine, homohistidine, N-acetyl-histidine, α-fluoromethyl-histidine, α-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine;
Xaa8 is Ala, Gly, Val, Leu, Ile, Lys, Aib, (1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
Xaa18 is Ser, Lys or Arg;
Xaa22 is Gly, Glu or Aib;
Xaa23 is Gln, Glu, Lys or Arg;
Xaa26 is Lys, Glu or Arg;
Xaa30 is Ala, Glu or Arg;
Xaa34 is Lys, Glu or Arg;
Xaa35 is Gly or Aib;
Xaa36 is Arg or Lys, amide or is absent;
Xaa37 is Gly, Ala, Glu or Lys, amide or is absent, and
(II) component (II) comprises:
C-terminally a peptide sequence of at least 9 amino acids,
wherein component (II) is a peptide sequence containing a sequence according to SEQ ID NO: 22 (RRDFPEEVAI) or a sequence having at least 80% sequence homology with SEQ ID NO: 22;
(A) preparing a core comprising a cross-linked polymer and biological cells capable of expressing and secreting a fusion peptide, wherein the fusion peptide comprises: components (I) and (II), wherein:
(I) component (I) comprises:
(a) N-terminally a GLP-1 (7-35, 7-36 or 7-37) sequence or a fragment or variant thereof; or
(b) a peptide comprising the sequence according to formula II:
(SEQ ID NO: 28)
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa16-Ser-
Xaa18-Xaa19-Xaa20-Glu-Xaa22-Xaa23-Ala-Xaa25-Xaa26-
Xaa27-Phe-Ile-Xaa30-Trp-Leu-Xaa33-Xaa34-Xaa35-
Xaa36-Xaa37,
wherein
Xaa7 is L-histidine, D-histidine, desamino-histidine, 2-aminohistidine, 3-hydroxy-histidine, homohistidine, N-acetyl-histidine, α-fluoromethyl-histidine, α-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine;
Xaa8 is Ala, Gly, Val, Leu, He, Lys, Aib, (1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
Xaa16 is Val or Leu;
Xaa18 is Ser, Lys or Arg;
Xaa19 is Tyr or Gln;
Xaa20 is Leu or Met;
Xaa22 is Gly, Glu or Aib;
Xaa23 is Gln, Glu, Lys or Arg;
Xaa25 is Ala or Val;
Xaa26 is Lys, Glu or Arg;
Xaa27 is Glu or Leu;
Xaa30 is Ala, Glu or Arg;
Xaa33 is Val or Lys;
Xaa34 is Lys, Glu, Asn or Arg;
Xaa35 is Gly or Aib;
Xaa36 is Arg, Gly or Lys or amide or absent;
Xaa37 is Gly, Ala, Glu, Pro, Lys, amide or is absent; or
(c) a peptide comprising a sequence according to formula III:
(SEQ ID NO: 29)
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-
Xaa18-Tyr-Leu-Glu-Xaa22-Xaa23-Ala-Ala-Xaa26-Glu-
Phe-Ile-Xaa30-Trp-Leu-Val-Xaa34-Xaa35-Xaa36-Xaa37,
wherein
Xaa7 is L-histidine, D-histidine, desamino-histidine, 2-aminohistidine, 3-hydroxy-histidine, homohistidine, N-acetyl-histidine, α-fluoromethyl-histidine, α-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine;
Xaa8 is Ala, Gly, Val, Leu, Ile, Lys, Aib, (1-aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl) carboxylic acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, or (1-aminocyclooctyl) carboxylic acid;
Xaa18 is Ser, Lys or Arg;
Xaa22 is Gly, Glu or Aib;
Xaa23 is Gln, Glu, Lys or Arg;
Xaa26 is Lys, Glu or Arg;
Xaa30 is Ala, Glu or Arg;
Xaa34 is Lys, Glu or Arg;
Xaa35 is Gly or Aib;
Xaa36 is Arg or Lys, amide or is absent;
Xaa37 is Gly, Ala, Glu or Lys, amide or is absent, and
(II) component (II) comprises:
C-terminally a peptide sequence of at least 9 amino acids,
wherein component (II) is a peptide sequence containing a sequence according to SEQ ID NO: 22 (RRDFPEEVAI) or a sequence having at least 80% sequence homology with SEQ ID NO: 22; and
(B) preparing at least one surface coating comprising a cross-linked polymer.
60 . A method according to claim 59 for producing a spherical or non-spherical microcapsule.
61 . A method for the treatment of diabetes mellitus type I or type II, insulin resistance, weight disorders and diseases or conditions associated thereto comprising administering to a subject in need thereof a pharmaceutical composition according to claim 58 .
62 . The method according to claim 61 , wherein the weight disorders or associated conditions include obesity, overweight-associated conditions, satiety deregulation, reduced plasma insulin levels, increased blood glucose levels, or reduced pancreatic beta cell mass.
63 . A method for the treatment of neurodegenerative disorders and diseases or conditions associated thereto comprising administering to a subject in need thereof a pharmaceutical composition according to claim 58 .
64 . The method according to claim 63 , wherein the neurodegenerative disorders and diseases or conditions associated thereto are selected from the group consisting of Alzheimer's disease (AD) and other central and peripheral neurodegenerative conditions selected from amyotrophic lateral sclerosis (ALS), Alexander disease, Alper's disease, Ataxia telangiectasia, Canavan disease, Cockayne syndrome, Creutzfeldt-Jakob disease, Multiple Sclerosis, Sandhoff disease, Pick's disease, Spinocerebellar Ataxia, Schilder's disease and Parkinson's disease, or stroke, intracerebral haemorrhage (ICH), subarachnoid haemorrhage, adenoleukodystrophy (x-ALD) or other leukodystrophies.
65 . A method for the treatment of disorders and diseases or conditions associated with apoptosis comprising administering to a subject in need thereof a pharmaceutical composition according to claim 64 .Join the waitlist — get patent alerts
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