Variants of tissue inhibitor or metalloprotienase type three (TIMP-3), compositions and methods

Patent metadata

Field	Value
Publication number	US-11149078-B2
Application number	US-201515547730-A
Country	US
Kind code	B2
Filing date	Aug 26, 2015
Priority date	Aug 27, 2014
Publication date	Oct 19, 2021
Grant date	Oct 19, 2021

Abstract

The application concerns tissue inhibitor of metalloproteinase 3 (TIMP-3) muteins, variants and derivatives, nucleic acids encoding them, and methods of making and using them; in particular, muteins of TIMP-3 with specific amino acid substitutions in order to introduce N-linked glycosylation sites.

First claim

What is claimed is:

1. An isolated Tissue Inhibitor of Metalloproteinase Type Three (TIMP-3) mutein having a mature region that is at least 90% identical in amino acid sequence to the mature region of TIMP-3 set forth in SEQ ID NO:2, selected from the group consisting of:
a) a TIMP-3 mutein having two or more pairs of mutations selected from the group consisting of K45N/V47T; K50N/V52T; P56N/G58T; H78N/Q80T; K94N/E96T; and D110N/K112T;
b) a TIMP-3 mutein having one or more pairs of mutations selected from the group consisting of K45N/V47T; K50N/V52T; P56N/G58T; H78N/Q80T; K94N/E96T; and D110N/K112T; and an additional mutation that is selected from the group consisting of R138T; G173T, and both R138T and G173T; and
c) the TIMP-3 mutein according to a) or b) that further comprises the mutation F57N.

2. A Tissue Inhibitor of Metalloproteinase Type Three (TIMP-3) mutein having a mature region that is at least 90% identical in amino acid sequence to amino acids 24-211 of SEQ ID NO:2, selected from the group consisting of:
a) a TIMP-3 mutein having two or more pairs of mutations selected from the group consisting of K45N/V47T; K50N/V52T; P56N/G58T; H78N/Q80T; K94N/E96T; and D110N/K112T;
b) a TIMP-3 mutein having one or more pairs of mutations selected from the group consisting of K45N/V47T; K50N/V52T; P56N/G58T; H78N/Q80T; K94N/E96T; and D110N/K112T; and an additional mutation that is selected from the group consisting of R138T; G173T, and both R138T and G173T; and c) the TIMP-3 mutein according to a) or b) that further comprises the mutation F57N.

3. The TIMP-3 mutein of claim 1 , comprising a group of mutations selected from the group consisting of:
(i) K45N/V47T, P56N/G58T, Q126N, and R138T;
(ii) K45N/V47T, P56N/G58T, K94N/E96T, and R138T;
(iii) K45N/V47T, P56N/G58T, R138T and G173T;
(iv) K45N/V47T, K94N/E96T, D110N/K112T, and F57N;
(v) K45N/V47T, K94N/E96T, F57N and R138T;
(vi) K45N/V47T, H78N/Q80T, K94N/E96T, R138T, and G173T;
(vii) K45N/V47T, K94N/E96T, D110N/K112T, and R138T;
(viii) K45N/V47T, K94N/E96T, D110N/K112T, and G173T;
(ix) K45N/V47T, K94N/E96T, R138T and G173T;
(x) K94N/E96T, D110N/K112T, K45S, F57N, and R138T;
(xi) H78N/Q80T, K94N/E96T, K45S, F57N and R138T;
(xii) K50N/V52T, P56N/G58T, K94N/E96T, D110N/K112T, R138T;
(xiii) K50N/V52T, H78N/Q80T, K94N/E96T, R138T and G173T;
(xiv) K50N/V52T, K94N/E96T, D110N/K112T, and R138T;
(xv) K50N/V52T, K94N/E96T, D110N/K112T, R138T and G173T;
(xvi) K50N/V52T, K94N/E96T, R138T and G173T;
(xvii) K50N/V52T, Q126N, R138T, and G173T;
(xviii) P56N/G58T, H78N/Q80T, K94N/E96T, and R138T;
(xix) P56N/G58T, K94N/E96T, Q126N and R138T;
(xx) P56N/G58T, K94N/E96T, D110N/K112T, and R138T;
(xxi) P56N/G58T, H78N/Q80T, K94N/E96T, and G173T;
(xxii) P56N/G58T, Q126N, R138T, and G173T;
(xxiii) H78N/Q80T, K94N/E96T, R138T and G173T;
(xxiv) H78N/Q80T, K94N/E96T, D110N/K112T, and R138T;
(xxv) K50N/V52T, D110N/K112T, R138T and G173T;
(xxvi) K45N/V47T, D110N/K112T, R138T and G173T;
(xxvii) H78N/Q80T, D110N/K112T, R138T and G173T;
(xxviii) K45N/V47T, K50N/V52T, H78N/Q80T, R138T;
(xxix) K45N/V47T, H78N/Q80T, D110N/K112T, and G173T;
(xxxx) K45N/V47, H78N/Q80T, R138T and G173T;
(xxxi) K50N/V52T, H78N/Q80T, K94N/E96T, and G173T;
(xxxii) K50N/V52T, H78N/Q80T, D110N/K112T, and R138T;
(xxxiii) K45N/V47T, K50N/V52T, H78N/Q80T, and D110N/K112T;
(xxxiv) K50N/V52T, H78N/Q80T, R138T and G173T;
(xxxv) K45N/V47T, H78N/Q80T, R138T and G173T;
(xxxvi) K45N/V47T, H78N/Q80T, and D110N/K112T, and R138T;
(xxxvii) K45N/V47T, K50N/V52T, H78N/Q80T, D110N/K112T, and G173T;
(xxxviii) K45N/V47T, K50N/V52T, H78N/Q80T, and R138T and G173T;
(xxxix) K45N/V47T, K50N/V52T, H78N/Q80T, K94N/E96T, and G173T;
(xl) K45N/V47T, H78N/Q80T, K94N/E96T, R138T and G173T;
(xli) K50N/V52T, H78N/Q80T, K94N/E96T, R138T and G173T;
(xlii) K45N/V47T, H78N/Q80T, and D110N/K112T, R138T and G173T;
(xliii) K50N/V52T, H78N/Q80T, D110N/K112T, R138T and G173T; and
(xliv) K45N/V52T, K50N/V52T, H78N/Q80T, D110N/K112T, and R138T.

4. The TIMP-3 mutein of claim 3 , comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 3-26.

5. The TIMP-3 mutein of claim 1 fused or conjugated to a moiety that extends half-life of a polypeptide.

6. The TIMP-3 mutein of claim 5 fused to an antibody, an Fc portion of an antibody, the heavy chain or light chain of an antibody, or human serum albumin.

7. The TIMP-3 mutein of claim 5 conjugated to polyethylene glycol.

8. An isolated nucleic acid that encodes a Tissue Inhibitor of Metalloproteinase Type Three (TIMP-3) mutein according to claim 1 .

9. An expression vector comprising the isolated nucleic acid of claim 8 .

10. An isolated host cell transformed or transfected with the expression vector of claim 9 .

11. A method of producing a recombinant Tissue Inhibitor of Metalloproteinase Type Three (TIMP-3) mutein comprising culturing the transformed or transfected host cell of claim 10 under conditions promoting expression of the TIMP-3 mutein, and recovering the TIMP-3 mutein.

12. A composition comprising the Tissue Inhibitor of Metalloproteinase Type Three (TIMP-3) mutein of claim 1 and a physiologically acceptable diluent, excipient or carrier.

13. A method of inhibiting cardiac extracellular matrix (ECM) degradation or adverse cardiac remodeling in a subject, the method comprising administering to a subject in need thereof an amount of composition of claim 12 effective to inhibit ECM degradation and/or adverse cardiac remodeling.

14. The method of claim 13 , wherein the subject has suffered a myocardial infarction.

15. The method of claim 13 , wherein the composition is administered via intracoronary administration or direct injection into the myocardium.

16. The isolated TIMP-3 mutein of claim 1 , comprising the mutations H78N/Q80T, K94N/E96T, and D110N/K112T.

17. The isolated TIMP-3 mutein of claim 16 , further comprising one or more mutations selected from the group consisting of K45N/V47T, K50N/V52T, and P56N/G58T.

18. The isolated TIMP-3 mutein of claim 16 , further comprising a mutation selected from the group consisting of R138T, G173T, and both R138T and G173T.

19. The isolated TIMP-3 mutein of claim 16 , further comprising the mutation F57N.

20. The isolated TIMP-3 mutein of claim 2 , comprising the mutations H78N/Q80T, K94N/E96T, and D110N/K112T.

21. The isolated TIMP-3 mutein of claim 20 , further comprising one or more mutations selected from the group consisting of K45N/V47T, K50N/V52T, and P56N/G58T.

22. The isolated TIMP-3 mutein of claim 20 , further comprising a mutation selected from the group consisting of R138T, G173T, and both R138T and G173T.

23. The isolated TIMP-3 mutein of claim 20 , further comprising the mutation F57N.

24. A composition comprising the Tissue Inhibitor of Metalloproteinase Type Three (TIMP-3) mutein of claim 16 and a physiologically acceptable diluent, excipient or carrier.

25. A composition comprising the TIMP-3 mutein of claim 20 and a physiologically acceptable diluent, excipient or carrier.

26. A method of inhibiting cardiac extracellular matrix (ECM) degradation or adverse cardiac remodeling in a subject, the method comprising administering to a subject in need thereof an amount of composition of claim 24 effective to inhibit ECM degradation and/or adverse cardiac remodeling.

27. A method of inhibiting cardiac extracellular mat

Assignees

• Amgen Inc

Inventors

• O'Neill Jason C
• Ketchem Randal R
• Lee Taeweon
• Chintalgattu Vishnu
• Stevens Jennitte Leann

Classifications

• A61P21/00

  Drugs for disorders of the muscular or neuromuscular system · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P9/04

  Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure · CPC title

• C07K14/8146 Primary

  Metalloprotease (E.C. 3.4.24) inhibitors, e.g. tissue inhibitor of metallo proteinase, TIMP · CPC title

• A61P33/00

  Antiparasitic agents · CPC title