Companion diagnostic for anti-hyaluronan agent therapy and methods of use thereof

• US 9,458,442 B2
• Filed: 08/14/2014
• Issued: 10/04/2016
• Est. Priority Date: 10/24/2011
• Status: Expired due to Fees

• Current Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Original Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Inventors: Jiang, Ping, Shepard, H. Michael, Huang, Lei
• Primary Examiner(s): Carlson, Karen Cochrane
• Application Number:
  US14/459,876
• Publication Number:
  US 20140348817A1
• Time in Patent Office: 782 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/735 containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)
• CPC Class Codes: C12N 9/2474 Hyaluronoglucosaminidase (3.2.1.35), i.e. hyaluronidase
• CPC Class Codes: G01N 2333/705 Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 2400/40 Glycosaminoglycans, i.e. GAG or mucopolysaccharides, e.g. chondroitin sulfate, dermatan sulfate, hyaluronic acid, heparin, heparan sulfate, and related sulfated polysaccharides
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: G01N 33/57488 involving compounds identifable in body fluids
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

Methods and diagnostic agents for identification of subjects for cancer treatment with an anti-hyaluronan agent, such as a hyaluronan-degrading enzyme, are provided. Diagnostic agents for the detection and quantification of hyaluronan in a biological sample and monitoring cancer treatment with an anti-hyaluronan agent, for example a hyaluronan-degrading enzyme, are provided. Combinations and kits for use in practicing the methods also are provided.

Claims

1. A nucleic acid molecule encoding a TSG-6 multimer, wherein:

the nucleic acid molecule encodes a fusion polypeptide containing a TSG-6 link module linked directly or indirectly via a linker to a multimerization domain; and

the multimerization domain is a polypeptide that interacts with itself to form a stable protein-protein interaction, whereby the encoded protein forms a multimer containing at least two TSG-6 link modules.

2. The nucleic acid molecule of claim 1, wherein the link module is the only TSG-6 portion of the fusion polypeptide.

3. The nucleic acid molecule of claim 1, wherein the encoded TSG-link module comprises the sequence of amino acids set forth in SEQ ID NO:

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 NO;

207, 360, 417 or 418 that specifically binds to hyaluronan (HA).

4. The nucleic acid molecule of claim 1, wherein the TSG-6 multimer has a binding affinity to hyaluronan (HA) with an association constant (Ka) of at least 1×

10⁷ M<sup>−

1</sup>.

5. The nucleic acid molecule of claim 1, wherein the encoded TSG-6 link module is modified to reduce or eliminate binding to heparin.

6. The nucleic acid molecule of claim 5, wherein the encoded 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.

7. The nucleic acid molecule of claim 6, wherein the amino acid replacement is to a non-basic amino acid residue selected from among Asp (D), Glu (E), Ser (S), Thr (T), Asn (N), Gln (Q), Ala (A), Val (V), Ile (I), Leu (L), Met (M), Phe (F), Tyr (Y) and Trp (W).

8. The nucleic acid molecule of claim 7, wherein the encoded TSG-6 link module comprises an amino acid replacement corresponding to amino acid replacement selected from among K20A, K34A and 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.

9. The nucleic acid molecule of claim 8, wherein the encoded TSG-6 link module comprises the sequence of amino acids 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.

10. The nucleic acid molecule of claim 1, wherein the encoded multimerization domain is selected from among an immunoglobulin constant region (Fc), a leucine zipper, complementary hydrophobic regions, complementary hydrophilic regions, compatible protein-protein interaction domains, free thiols that form 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.

11. The nucleic acid molecule of claim 10, wherein the encoded multimerization domain is an Fc domain or a variant thereof that effects multimerization.

12. The nucleic acid molecule of claim 11, wherein the encoded TSG-6 multimer comprises 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 amino acids set forth in SEQ ID NO;

212 or 215 and specifically binds HA.

13. The nucleic acid molecule of claim 11, comprising the sequence of nucleotides set forth in SEQ ID NO:

211, 214 or 217, or a sequence of nucleotides that exhibits at least 85% sequence identity to the sequence of nucleotides set forth in SEQ ID NO;

211, 214 or 217.

14. The nucleic acid molecule of claim 11, consisting of the sequence of nucleotides set forth in SEQ ID NO:

211, 214 or 217, or a sequence of nucleotides that exhibits at least 85% sequence identity to the sequence of nucleotides set forth in SEQ ID NO;

211, 214 or 217.

15. A cell, comprising the nucleic acid molecule of claim 1.

16. A vector, comprising the nucleic acid molecule of claim 1.

17. A method of producing a TSG-6 multimer, comprising:

introducing the nucleic acid molecule of claim 1 into a cell;

culturing the cell under conditions whereby the fusion polypeptide is expressed by the cell; and

recovering the TSG-6 multimer.