Human ctla4 mutants and use thereof

Patent metadata

Field	Value
Publication number	US-2016017018-A1
Application number	US-201414773056-A
Country	US
Kind code	A1
Filing date	Mar 5, 2014
Priority date	Mar 7, 2013
Publication date	Jan 21, 2016
Grant date	—

Abstract

Mutant forms of human CTLA4, and their use, e.g., in xenotransplantation.

First claim

1 . A mutant human Cytotoxic T-Lymphocyte-Associated Protein 4 (CTLA4) protein comprising a sequence that is at least 80% identical to the full length of amino acids 39-152 of SEQ ID NO: 1, wherein the amino acid at position 134 is a Leucine.

2 . The mutant human CTLA4 protein of claim 1 , wherein
(i) the amino acid at position 113 is not Asparagine (N), and/or the amino acid at position 115 is neither Threonine (T) nor Serine (S); and (ii) the amino acid at position 145 is not Asparagine (N), and/or the amino acid at position 147 is neither Threonine (T) nor Serine (S).

3 . The mutant human CTLA4 protein of claim 2 , wherein the amino acids at positions 113 and 145 are Alanine (A) or Glycine (G).

4 . The mutant human CTLA4 protein of claim 1 , which is at least 80% identical to the full length of amino acids 36-161 of SEQ ID NO: 1, wherein the amino acid at position 134 is a Leucine.

5 . A mutant human CTLA4 fusion protein comprising:
a first part comprising a mutant human Cytotoxic T-Lymphocyte-Associated Protein 4 (CTLA4) comprising a sequence that is at least 80% identical to the full length of amino acids 39-152 of SEQ ID NO: 1, wherein the amino acid at position 134 is a Leucine, and wherein optionally (i) the amino acid at position 113 is not Asparagine (N), and/or the amino acid at position 115 is neither Threonine (T) nor Serine (S); and (ii) the amino acid at position 145 is not Asparagine (N), and/or the amino acid at position 147 is neither Threonine (T) nor Serine (S); and a second part comprising an Fc region of a human immunoglobulin.

6 . The fusion protein of claim 4 , wherein the second part comprises the Fc region of human IgG.

7 . The fusion protein of claim 4 , wherein the a mutant human Cytotoxic T-Lymphocyte-Associated Protein 4 (CTLA4) is at least 80% identical to the full length of amino acids 36-161 of SEQ ID NO: 1, wherein the amino acid at position 134 is a Leucine.

8 . A codon-optimized nucleic acid molecule encoding the mutant human CTLA-4 of claim 1 , preferably optimized for expression in a methylotropic yeast, preferably of the species Pichia Pastoris.

9 . A vector comprising the nucleic acid molecule of claim 8 .

10 . A host cell expressing the nucleic acid molecule of claim 8 .

11 . The host cell of claim 10 , wherein the host cell is a methylotropic yeast.

12 . The host cell of claim 11 , wherein the host cell is a cell of the species Pichia Pastoris.

13 . A pharmaceutical composition comprising the mutant human CTLA-4 of claim 1 , and a physiologically acceptable carrier.

14 . A method of inducing tolerance in a human subject who has undergone or will undergo an organ transplantation procedure with a porcine organ, the method comprising administering to the subject a therapeutically effective amount of the fusion protein of claim 5 .

15 . (canceled)

16 . (canceled)

17 . A method of producing a mutant human CTLA-4, the method comprising: expressing a mutant human CTLA-4 a sequence that is at least 80% identical to the full length of amino acids 39-152 of SEQ ID NO: 1, wherein the amino acid at position 134 is a Leucine, and wherein optionally
(i) the amino acid at position 113 is not Asparagine (N), and/or the amino acid at position 115 is neither Threonine (T) nor Serine (S); and (ii) the amino acid at position 145 is not Asparagine (N), and/or the amino acid at position 147 is neither Threonine (T) nor Serine (S), in a methylotropic yeast; and substantially purifying the human CTLA-4, thereby producing the composition.

18 . The method of claim 17 , wherein the methylotropic yeast is of the species Pichia Pastoris.

19 . The method of claim 17 , wherein the mutant human CTLA4 is at least 80% identical to the full length of amino acids 36-161 of SEQ ID NO:1, wherein the amino acid at position 134 is a Leucine.

20 . A codon-optimized nucleic acid molecule encoding the mutant human CTLA-4 fusion protein of claim 5 , preferably optimized for expression in a methylotropic yeast, preferably of the species Pichia Pastoris.

21 . A pharmaceutical composition comprising the mutant human CTLA-4 fusion protein of claim 5 , and a physiologically acceptable carrier.

Assignees

• Gen Hospital Corp

Inventors

• Wang Zhirui
• Huang Christene A
• Sachs David H

Classifications

• A61K38/00

  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 title

• A61K39/001

  Preparations to induce tolerance to non-self, e.g. prior to transplantation · CPC title

• C07K16/18

  against material from animals or humans · CPC title

• A61K39/3955

  against proteinaceous materials, e.g. enzymes, hormones, lymphokines · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title