Methods of treatment of collagen-mediated diseases and conditions

• US 9,833,498 B2
• Filed: 03/06/2009
• Issued: 12/05/2017
• Est. Priority Date: 03/06/2008
• Status: Active Grant

• Current Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Original Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Inventors: Keller, Gilbert, Frost, Gregory I.
• Primary Examiner(s): Swope, Sheridan
• Application Number:
  US12/381,063
• Publication Number:
  US 20100003237A1
• Time in Patent Office: 3,196 Days
• CPC Class Codes: A61K 38/4813 Exopeptidases (3.4.11. to 3.4.19)
• CPC Class Codes: A61K 38/482 Serine endopeptidases (3.4.21)
• CPC Class Codes: A61K 38/4873 Cysteine endopeptidases (3.4.22), e.g. stem bromelain, papain, ficin, cathepsin H
• CPC Class Codes: A61K 38/488 Aspartic endopeptidases (3.4.23), e.g. pepsin, chymosin, renin, cathepsin E
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 41/00 Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 9/6472 Cysteine endopeptidases (3.4.22)
• CPC Class Codes: C12N 9/6491 Matrix metalloproteases [MMP's], e.g. interstitial collagenase (3.4.24.7); Stromelysins (3.4.24.17; 3.2.1.22); Matrilysin (3.4.24.23)
• CPC Class Codes: C12Y 304/22015 Cathepsin L (3.4.22.15)
• CPC Class Codes: C12Y 304/24007 Interstitial collagenase (3.4.24.7), i.e. matrix metalloprotease 1 or MMP1

Abstract

Methods and combinations are provided for controlling the duration of action, in vivo, of matrix-degrading enzymes. The methods and combinations permit temporary in-vivo activation of matrix-degrading enzymes upon administration to the extra cellular matrix (or “ECM”). Matrix-degrading enzymes having a controlled duration of action can be used to treat ECM-mediated diseases or disorders characterized by increased deposition or accumulation of one or more ECM components.

Claims

1. A method for treating a collagen-mediated disease or condition of the sub-epidermal extracellular matrix (ECM), comprising sub-epidermally delivering a composition into the sub-epidermal ECM of a subject, wherein:

sub-epidermal delivery to the ECM is selected from among subcutaneous and intralesional administration;

the composition comprises a protein comprising an enzyme that consists of the sequence of amino acids set forth in SEQ ID NO;

1, or an enzyme that consists of a sequence of amino acids that exhibits at least 90% sequence identity to the sequence of amino acids set forth in SEQ ID NO;

1 and has cathepsin L activity to cleave type I collagen;

the disease or condition is associated with accumulation of fibrous tissue rich in type I collagen under the epidermis of the skin;

the disease or condition is treated by degrading the type I collagen component of the ECM in the fibrous tissue under the epidermis of the skin; and

the pH of the composition is acidic, whereby the enzyme is active in the ECM upon administration, but is inactivated by the higher pH of the ECM whereby it is active for a limited time after administration so that the collagen component in the sub-epidermal ECM is degraded for a limited time to thereby treat the disease or condition.

2. The method of claim 1, wherein delivery is effected by injection.

3. The method of claims 1, wherein the enzyme in the composition is human cathepsin L.

4. The method of claim 1, wherein the enzyme is a single chain form.

5. The method of claim 1, wherein the enzyme is a two chain form.

6. The method of claim 1, wherein the enzyme consists of:

a heavy chain consisting of the sequence of amino acids set forth as amino acids 1-175 of SEQ ID NO;

1 or consisting of a sequence that exhibits at least 90% sequence identity to the sequence of amino acids 1 -175 of SEQ ID NO;

1; and

a light chain consisting of the sequence of amino acids set forth as amino acids 179-220 of SEQ ID NO;

1 or consisting of a sequence that exhibits at least 90% sequence identity to the sequence of amino acids 179-220 of SEQ ID NO;

1.

8. The method of claim 1, wherein the enzyme consists of the sequence of amino acids set forth in SEQ ID NO:

1.

9. The method of claim 1, wherein delivery is effected by subcutaneous administration, intralesional administration or intradermal administration.

10. The method of claim 1, wherein delivery is effected by subcutaneous administration.

11. The method of claim 1, wherein the pH of the composition is within a range of about 3 to 6.5.

12. The method of claim 1, wherein the pH of the composition is within a range of about 4 to 6.

13. The method of claim 1, wherein the pH of the composition is within a range of about 5 to 6.

14. The method of claim 1, wherein the composition comprises a buffer containing an acid selected from among 2-(N-morpholino)ethanesulfonic acid (MES), acetic acid, citric acid, succinic acid, lactic acid, maleic acid, glycine-hydrochloric acid, citric phosphate and histidine.

15. The method of claim 1, wherein the amount of the enzyme in the composition is between about 10 μ

g and 100 mg.

16. The method of claim 1, wherein the collagen-mediated disease or condition is cellulite.

17. The method of claim 1, wherein the composition comprising the protein is prepared by a method comprising:

providing a lyophilized preparation containing the protein; and

adding an acidic buffered solution at the acidic pH to the lyophilized preparation, thereby generating an acidic pH composition containing the protein.

18. The method of claim 1, wherein the enzyme in the composition is a mature enzyme that is generated from a zymogen by activation cleavage to cleave the propeptide, thereby producing the mature enzyme.

19. The method of claim 18, wherein the cleaved propeptide is removed from the composition.

20. The method of claim 18, wherein the zymogen consists of:

the sequence of amino acids set forth as amino acids 18-333 of SEQ ID NO;

62;

or

a sequence of amino acids that exhibits at least 90 % sequence identity to the sequence of amino acids set forth as amino acids 18-333 of SEQ ID NO;

62.

21. The method of claim 20, wherein the zymogen consists of the sequence of amino acids set forth as amino acids 18-333 of SEQ ID NO:

62.

22. The method of claim 1, wherein the protein consists of an enzyme that consists of the sequence of amino acids set forth in SEQ ID NO:

1, or a sequence of amino acids that exhibits at least 90% sequence identity to the sequence of amino acids set forth in SEQ ID NO;

1 and has cathepsin L activity to cleave type I collagen.

23. The method of claim 1, wherein the protein consists of an enzyme that consists of the sequence of amino acids set forth in SEQ ID NO:

1.

7. A method for treating a collagen-mediated disease or condition of the sub-epidermal extracellular matrix (ECM), comprising sub-epidermally delivering into the sub-epidermal ECM of a subject a composition comprising a cathepsin L enzyme, wherein:

sub-epidermal delivery to the ECM is selected from among subcutaneous and intralesional administration;

the cathepsin L enzyme comprises the sequence of amino acids set forth in SEQ ID NO;

1;

the disease or condition is associated with accumulation of fibrous tissue rich in type I collagen under the epidermis of the skin;

the disease or condition is treated by degrading the type I collagen component of the ECM in the fibrous tissue under the epidermis of the skin; and

the pH of the composition containing the cathepsin L enzyme is acidic, whereby the cathepsin L is active in the ECM upon administration, but is inactivated by the higher pH of the ECM whereby it is active for a limited time after administration so that the collagen component in the sub-epidermal ECM is degraded for a limited time to thereby treat the disease or condition.