Title:
FUSION PROTEIN FOR TREATING MYASTHENIA GRAVIS
Document Type and Number:
WIPO Patent Application WO/2022/260579
Kind Code:
A3
Abstract:
The invention relates to a method of producing a fusion protein between an extracellular domain of neuronal acetylcholine receptor subunit alpha 1 (nAChR&1) and a solubility enhancing peptide. The method comprises solubilizing inclusion bodies comprising the fusion protein in a solubilization solution having a pH of at least 11 to form solubilized fusion proteins. The method also comprises diluting the solubilized fusion protein in a refolding solution having a pH of no more than 9 to form a refolded monomeric form of the fusion protein.
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Inventors:
ANDERSSSON JAN-OLOF (SE)
CHRISTISON RICHARD IAN (SE)
FRIBERT CHARLOTTE (SE)
LÖWENADLER BJÖRN (SE)
SJÖHAMN JENNIE (SE)
CHRISTISON RICHARD IAN (SE)
FRIBERT CHARLOTTE (SE)
LÖWENADLER BJÖRN (SE)
SJÖHAMN JENNIE (SE)
Application Number:
PCT/SE2022/050556
Publication Date:
January 26, 2023
Filing Date:
June 07, 2022
Export Citation:
Assignee:
TOLERANZIA AB (SE)
International Classes:
C07K14/725; A61K38/00; A61P25/00
Other References:
ZISIMOPOULOU P ET AL: "Towards antigen-specific apheresis of pathogenic autoantibodies as a further step in the treatment of myasthenia gravis by plasmapheresis", JOURNAL OF NEUROIMMUNOLOGY, ELSEVIER SCIENCE PUBLISHERS BV, NL, vol. 201-202, 15 September 2008 (2008-09-15), pages 95 - 103, XP025495583, ISSN: 0165-5728, [retrieved on 20080729], DOI: 10.1016/J.JNEUROIM.2008.06.020
LAZARIDIS KONSTANTINOS ET AL: "Expression of human AChR extracellular domain mutants with improved characteristics", INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, vol. 63, 15 November 2013 (2013-11-15), NL, pages 210 - 217, XP055963555, ISSN: 0141-8130, DOI: 10.1016/j.ijbiomac.2013.11.003
THEODOROS TSOULOUFIS ET AL: "Reconstitution of conformationally dependent epitopes on the N-terminal extracellular domain of the human muscle acetylcholine receptor -alpha- subunit expressed in Escherichia coli: implications for myasthenia gravis therapeutic approaches", INTERNATIONAL IMMUNOLOGY, OXFORD UNIVERSITY PRESS, GB, vol. 12, no. 9, 1 September 2000 (2000-09-01), pages 1255 - 1265, XP008148987, ISSN: 0953-8178, DOI: 10.1093/INTIMM/12.9.1255
SCHRATTENHOLZ A ET AL: "Expression and renaturation of the N-terminal extracellular domain of Torpedo nicotinic acetylcholine receptor alpha-subunit", JOURNAL OF BIOLOGICAL CHEMISTRY, AMERICAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY, US, vol. 273, no. 49, 4 December 1998 (1998-12-04), pages 32393 - 32399, XP002174383, ISSN: 0021-9258, DOI: 10.1074/JBC.273.49.32393
LIXI NIU ET AL: "Potential roles of recombinant acetylcholine receptor subunit 1 211 in immunoadsorbent and DNA immunization", JOURNAL OF IMMUNOLOGICAL METHODS, ELSEVIER SCIENCE PUBLISHERS B.V.,AMSTERDAM, NL, vol. 372, no. 1, 5 April 2011 (2011-04-05), pages 14 - 21, XP028285036, ISSN: 0022-1759, [retrieved on 20110720], DOI: 10.1016/J.JIM.2011.04.015
PSARIDI-LINARDAKI L ET AL: "Expression of soluble ligand- and antibody-binding extracellular domain of human muscle acetylcholine receptor alpha subunit in yeast Pichia pastoris. Role of glycosylation in alpha-bungarotoxin binding", JOURNAL OF BIOLOGICAL CHEMISTRY, AMERICAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY, US, vol. 277, no. 30, 28 July 2002 (2002-07-28), pages 26980 - 26986, XP002225379, ISSN: 0021-9258, DOI: 10.1074/JBC.M110731200
SUN CHENJING ET AL: "Experimental study Improved methodology to obtain large quantities of correctly folded recombinant N-terminal extracellular domain of the human muscle acetylcholine receptor for inducing experimental autoimmune myasthenia gravis in rats", ARCHIVES OF MEDICAL SCIENCE, vol. 2, 12 August 2013 (2013-08-12), pages 389 - 395, XP055964435, ISSN: 1734-1922, DOI: 10.5114/aoms.2013.36921
LAZARIDIS KONSTANTINOS ET AL: "Expression of human AChR extracellular domain mutants with improved characteristics", INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, vol. 63, 15 November 2013 (2013-11-15), NL, pages 210 - 217, XP055963555, ISSN: 0141-8130, DOI: 10.1016/j.ijbiomac.2013.11.003
THEODOROS TSOULOUFIS ET AL: "Reconstitution of conformationally dependent epitopes on the N-terminal extracellular domain of the human muscle acetylcholine receptor -alpha- subunit expressed in Escherichia coli: implications for myasthenia gravis therapeutic approaches", INTERNATIONAL IMMUNOLOGY, OXFORD UNIVERSITY PRESS, GB, vol. 12, no. 9, 1 September 2000 (2000-09-01), pages 1255 - 1265, XP008148987, ISSN: 0953-8178, DOI: 10.1093/INTIMM/12.9.1255
SCHRATTENHOLZ A ET AL: "Expression and renaturation of the N-terminal extracellular domain of Torpedo nicotinic acetylcholine receptor alpha-subunit", JOURNAL OF BIOLOGICAL CHEMISTRY, AMERICAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY, US, vol. 273, no. 49, 4 December 1998 (1998-12-04), pages 32393 - 32399, XP002174383, ISSN: 0021-9258, DOI: 10.1074/JBC.273.49.32393
LIXI NIU ET AL: "Potential roles of recombinant acetylcholine receptor subunit 1 211 in immunoadsorbent and DNA immunization", JOURNAL OF IMMUNOLOGICAL METHODS, ELSEVIER SCIENCE PUBLISHERS B.V.,AMSTERDAM, NL, vol. 372, no. 1, 5 April 2011 (2011-04-05), pages 14 - 21, XP028285036, ISSN: 0022-1759, [retrieved on 20110720], DOI: 10.1016/J.JIM.2011.04.015
PSARIDI-LINARDAKI L ET AL: "Expression of soluble ligand- and antibody-binding extracellular domain of human muscle acetylcholine receptor alpha subunit in yeast Pichia pastoris. Role of glycosylation in alpha-bungarotoxin binding", JOURNAL OF BIOLOGICAL CHEMISTRY, AMERICAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY, US, vol. 277, no. 30, 28 July 2002 (2002-07-28), pages 26980 - 26986, XP002225379, ISSN: 0021-9258, DOI: 10.1074/JBC.M110731200
SUN CHENJING ET AL: "Experimental study Improved methodology to obtain large quantities of correctly folded recombinant N-terminal extracellular domain of the human muscle acetylcholine receptor for inducing experimental autoimmune myasthenia gravis in rats", ARCHIVES OF MEDICAL SCIENCE, vol. 2, 12 August 2013 (2013-08-12), pages 389 - 395, XP055964435, ISSN: 1734-1922, DOI: 10.5114/aoms.2013.36921
Attorney, Agent or Firm:
BARKER BRETTELL SWEDEN AB (SE)
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