Hypoxia and hyaluronan and markers thereof for diagnosis and monitoring of diseases and conditions and related methods

Patent metadata

Field	Value
Publication number	US-9278124-B2
Application number	US-201313998269-A
Country	US
Kind code	B2
Filing date	Oct 16, 2013
Priority date	Oct 16, 2012
Publication date	Mar 8, 2016
Grant date	Mar 8, 2016

Abstract

Provided herein are diagnostic methods for identifying subjects susceptible to treatment with a hypoxia-activated agent, and related methods. Also provided herein are methods of monitoring treatments with anti-hyaluronan agents, and related methods.

First claim

The invention claimed is:

1. A method of treating a subject having a hypoxia-related disease or condition, comprising:
a) measuring the level or amount of hyaluronan (HA) in a sample from a subject, whereby if the HA is at or above a predetermined level, the subject is susceptible to treatment with a hypoxia-activated agent, wherein:
the HA is measured by detecting binding of a HA-binding protein (HABP) to the sample, and
the HABP comprises a Tumor necrosis factor-Stimulated Gene-6 (TSG-6) link module (LM) or a sufficient portion thereof that specifically binds to HA;
b) selecting a susceptible subject for treatment with a hyaluronidase and a hypoxia-activated agent; and
c) administering a soluble hyaluronidase and a therapeutically effective amount of a hypoxia-activated agent to the subject, wherein the soluble hyaluronidase and the hypoxia-activated agent are administered simultaneously, sequentially or intermittently in any order, to thereby treat the subject.

2. The method of claim 1 , wherein the hypoxia-related disease or condition is a hyperproliferative disease or condition.

3. The method of claim 1 , wherein the hypoxia-related disease or condition is cancer, angiogenesis or an angiogenesis related disorder.

4. The method of claim 1 , wherein the sample is selected from among a tissue, cell and bodily fluid.

5. The method of claim 1 , wherein the sample is from a tumor.

6. The method of claim 1 , wherein the predetermined level is the level or amount of the HA in a control or reference sample.

7. The method of claim 6 , wherein the control or reference sample is selected from among:
a) an analogous sample from a subject who does not have a hypoxia-related disease or condition;
b) an analogous sample from a subject known to express low hyaluronan in the sample; and
c) a cell line.

8. The method of claim 1 , wherein:
the predetermined level is the mean or median level or amount of HA in a sample from a subject who does not have a hypoxia-related disease or condition; or
the predetermined level is the mean or median level or amount of HA in a sample from a subject known to have a hypoxia-related disease or condition.

9. The method of claim 1 , wherein the subject is selected and treated if the level or amount is elevated at least 0.5-fold, 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold or more compared to the predetermined level.

10. The method of claim 1 , wherein the sample is a bodily fluid that is plasma and the predetermined level of HA is at least or above 0.010 μg HA/mL.

11. The method of claim 1 , wherein the sample is a tumor and a subject is selected for treatment if moderate to high hyaluronan is measured.

12. The method of claim 11 , wherein moderate to high hyaluronan is measured if hyaluronan is present on at least 10%, 10% to 25%, or greater than 25% of the tumoral area.

13. The method of claim 1 , wherein the TSG-6-LM has the sequence of amino acids set forth in SEQ ID NOS: 207, 360, 417 or 418, or a sequence of amino acids comprising at least 85% amino acid sequence identity to the sequence of amino acids set forth in SEQ ID NOS: 207, 360, 417 or 418 and specifically binds HA.

14. The method of claim 1 , wherein the TSG-6 link module is modified to reduce or eliminate binding to heparin.

15. The method of claim 14 , wherein the TSG-6 link module comprises an amino acid replacement at an amino acid position corresponding to amino acid residue 20, 34, 41, 54, 56, 72 or 84 set forth in SEQ ID NO:360, whereby a corresponding amino acid residue is identified by alignment to a TSG-6-LM set forth in SEQ ID NO:360.

16. The method of claim 15 , wherein TSG-6 link module comprises an amino acid replacement corresponding to amino acid replacement K20A, K34A and/or K41A in a TSG-6-LM set forth in SEQ ID NO:360 or the replacement at the corresponding residue in another TSG-6-LM.

17. The method of claim 16 , wherein the HABP comprises a link module set forth in SEQ ID NO:361 or 416 or a sequence of amino acids comprising at least 85% amino acid sequence identity to the sequence of amino acids set forth in SEQ ID NO:361 or 416 that specifically binds HA.

18. The method of claim 1 , wherein the HABP is a multimer comprising a first HA-binding domain linked directly or indirectly via a linker to a multimerization domain and a second HA-binding domain linked directly or indirectly via a linker to a multimerization domain and wherein the first and second HA-binding domain each is a TSG-6 link module (LM), a variant thereof or a sufficient portion thereof that specifically binds to HA.

19. The method of claim 18 , wherein the TSG-6-LM comprises the sequence of amino acids set forth in SEQ ID NOS: 207, 360, 361, 416, 417 or 418 or a sequence of amino acids comprising at least 85% amino acid sequence identity to the sequence of amino acids set forth in SEQ ID NOS: 207, 360, 361, 416, 417 or 418 that specifically binds HA.

20. The method of claim 18 , wherein the multimerization domain is selected from among an immunoglobulin constant region (Fe), a leucine zipper, complementary hydrophobic regions, complementary hydrophilic regions, compatible protein-protein interaction domains, free thiols that forms an intermolecular disulfide bond between two molecules, and a protuberance-into-cavity and a compensatory cavity of identical or similar size that form stable multimers.

21. The method of claim 18 , wherein the multimerization domain is an Fc domain or a variant thereof that effects multimerization.

22. The method of claim 21 , wherein the HABP is a fusion protein that contains a TSG-6 link module and an immunoglobulin Fc domain.

23. The method of claim 22 , wherein the HABP is TSG-6-LM-Fc and comprises the sequence of amino acids set forth as amino acids 21-349 of SEQ ID NO:212 or 215 or a sequence of amino acids that exhibits at least 85% sequence identity to amino acids 21-349 of SEQ ID NO: 212 or 215 and specifically binds HA.

24. The method of claim 22 , wherein the HABP is TSG-6-LM-Fc and the polypeptide is encoded by a nucleic acid molecule that encodes the sequence of amino acids set forth in SEQ ID NO:212 or 215 or a sequence of amino acids that exhibits at least 85% amino acid sequence identity to SEQ ID NO:212 or 215 and specifically binds HA.

25. The method of claim 1 , wherein the hypoxia-activated agent is a hypoxia-activated prodrug.

26. The method of claim 25 , wherein the hypoxia-activated prodrug comprises a bioreductive group selected from among a quinone, aromatic N-oxide, aliphatic N-oxide, nitroheterocyclic compound and transition-metal complex.

27. The method of claim 25 , wherein the hypoxia-activated prodrug comprises an anti-neoplastic agent.

28. The method of claim 27 , wherein the anti-neoplastic agent is a pan-Her inhibitor.

29. The method of claim 25 , wherein the hypoxia-activated prodrug is selected from among a mitomycin C, porfiromycin, cyclopropamitosene, diaziquone, streptonigrin, EO9, RH1, tirapazamine, CEN-209, AQ4N, Nitracrine N-Oxide, PR-104, SN28343, SN29303, SN29730, KS119W, NLCQ-1, RSU1069, RB6145, CB1954, SN23862, SN24771, TH-281, TH-308, TH-302, TH1332, TH1431, SN29966, SN32807, PR-509 or PR-610 and derivatives or analogs thereof.

30. The method of claim 25 , wherein the hypoxia-activated prodrug is a conjugate comprising a hypoxia-activ

Assignees

• Shepard H Michael
• Li Xiaoming
• Thompson Curtis
• Halozyme Inc

Inventors

• Shepard H Michael
• Li Xiaoming
• Thompson Curtis

Classifications

• G01N33/575

  for cancer · CPC title

• A61K38/31

  Somatostatins · CPC title

• A61K31/407 Primary

  condensed with other heterocyclic ring systems, e.g. ketorolac, physostigmine · CPC title

• A61K31/4709

  Non-condensed quinolines and containing further heterocyclic rings · CPC title

• A61K31/53

  having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine (melarsoprol A61K31/555 {; with four nitrogen atoms A61K31/495}) · CPC title