Modified hyaluronidases and uses in treating hyaluronan-associated diseases and conditions

Abstract

Provided are combinations, compositions and kits containing a hyaluronan degrading enzyme, such as a soluble hyaluronidase, for treatment of hyaluronan-associated conditions, diseases and disorders. In one example, the products include an additional agent or treatment. Such products can be used in methods for administering the products to treat the hyaluronan-associated diseases and conditions, for example, hyaluronan-associated cancers, for example, hyaluronan-rich tumors. The methods include administration of the hyaluronan degrading enzyme composition alone or in combination with other treatments. Also provided are methods and compositions for providing sustained treatment effects in hyaluronan-associated diseases and conditions.

1 . A method for treating a tumor with a polymer-conjugated soluble human PH20 hyaluronidase in a subject having cancer, comprising:
 a) measuring the expression or level of hyaluronan in a tumor sample from the subject; and the tumor sample is a tumor biopsy sample;   b) determining the level of expression of hyaluronan in the tumor biopsy sample;   c) identifying, for treatment, the subject, wherein hyaluronan is expressed in at least 30% of the tumoral area in the tumor biopsy sample from the subject; and then   d) administering to the subject, the polymer-conjugated soluble hyaluronidase.   
     
     
         2 . The method of  claim 1 , wherein the polymer-conjugated soluble human PH20 is a PEGylated PH20 (PEGPH20). 
     
     
         3 . The method of  claim 1 , wherein the cancer is selected from among ovarian cancer, in situ carcinoma (ISC), squamous cell carcinoma (SCC), prostate cancer, gastric cancer, non-small cell lung cancer, breast cancer, brain cancer and colon cancer. 
     
     
         4 . The method of  claim 1 , further comprising administering a second agent that is an anti-cancer agent or treatment for treating the tumor in the subject. 
     
     
         5 . The method of  claim 1 , wherein the polymer-conjugated soluble hyaluronidase is a C-terminally truncated human PH20 hyaluronidase that lacks all or a portion of a C-terminal glycosylphosphatidylinositol (GPI) anchor. 
     
     
         6 . The method of  claim 1 , wherein the polymer-conjugated soluble PH20 hyaluronidase is a polypeptide consisting of an amino acid sequence having at least 98% amino acid sequence identity with the amino acid sequence of SEQ ID NO:48, wherein the soluble PH20 hyaluronidase is N-glycosylated and neutral active. 
     
     
         7 . The method of  claim 1 , wherein the polymer-conjugated soluble PH20 hyaluronidase consists of the amino acid sequence set forth as amino acids 36-467, 36-468, 36-469, 36-470, 36-471, 36-472, 36-473, 36-474, 36-475, 36-476, 36-477, 36-478, 36-479, 36-480, 36-481, 36-482, or 36-483 of the amino acid sequence of SEQ ID NO:1, or an amino acid sequence having at least 95% amino acid sequence identity with the amino acid sequence set forth as amino acids 36-467, 36-468, 36-469, 36-470, 36-471, 36-472, 36-473, 36-474, 36-475, 36-476, 36-477, 36-478, 36-479, 36-480, 36-481, 36-482, or 36-483 of the amino acid sequence of SEQ ID NO:1. 
     
     
         8 . The method of  claim 1 , wherein the polymer-conjugated soluble hyaluronidase is a PEGylated soluble PH20 hyaluronidase, wherein the soluble PH20 hyaluronidase is selected from among a polypeptide consisting of the amino acid sequence set forth in SEQ ID NOs: 4-9 and 47-48, or variants thereof that have at least 98% amino acid sequence identity to the amino acid sequence of SEQ ID NO:48. 
     
     
         9 . The method of  claim 1 , wherein:
 the polymer-conjugated soluble hyaluronidase is PEGylated; and   PEGylation results from reaction with a PEG reagent selected from methoxy-poly(ethylene glycol)-succinimidyl butanoate (mPEG-SBA) (5 kDa); methoxy-poly(ethylene glycol)-succinimidyl butanoate (mPEG-SBA) (20 kDa); methoxy-poly(ethylene glycol)-succinimidyl butanoate (mPEG-SBA) (30 kDa); methoxy-poly(ethylene glycol)-succinimidyl α-methylbutanoate (mPEG-SMB) (20 kDa); methoxy-poly(ethylene glycol)-succinimidyl α-methylbutanoate (mPEG-SMB) (30 kDa); methoxy-poly(ethylene glycol)-butyraldehyde (mPEG-butyraldehyde) (30 kDa), methoxy-poly(ethylene glycol)-succinimidyl propionate (mPEG-SPA) (20 kDa); methoxy-poly(ethylene glycol)-succinimidyl propionate (mPEG-SPA) (30 kDa); (methoxy-poly(ethylene glycol)) 2-N-hydroxysuccinimide ester (mPEG2-NHS) (10 kDa branched); (methoxy-poly(ethylene glycol)) 2-N-hydroxysuccinimide ester (mPEG2-NHS) (20 kDa branched); (methoxy-poly(ethylene glycol)) 2-N-hydroxysuccinimide ester (mPEG2-NHS) (40 kDa branched); (methoxy-poly(ethylene glycol)) 2-N-hydroxysuccinimide ester (mPEG2-NHS) (60 kDa branched); biotin-poly(ethylene glycol)-N-hydroxysuccinimide ester (biotin-PEG-NHS) (5 kDa biotinylated); poly(ethylene glycol)-p-nitrophenyl carbonate (PEG-p-nitrophenyl-carbonate) (30 kDa); and poly(ethylene glycol)-propionaldehyde (PEG-propionaldehyde) (30 kDa).   
     
     
         10 . The method of  claim 1 , wherein:
 the polymer-conjugated soluble hyaluronidase is PEGylated; and   the PEG is a branched or linear PEG.   
     
     
         11 . The method of  claim 1 , wherein:
 the polymer-conjugated soluble hyaluronidase is PEGylated; and   the PEG has a mass of at least about 30 kilodaltons.   
     
     
         12 . The method of  claim 1 , wherein the polymer-conjugated soluble PH20 hyaluronidase is administered intravenously, subcutaneously, intraperitoneally, or intra-tumorally. 
     
     
         13 . The method of  claim 1 , wherein the polymer-conjugated soluble PH20 hyaluronidase is administered at a dose of 10 to 50,000,000 Units. 
     
     
         14 . A method for treating a disease or condition in which a hyaluronidase substrate accumulates, comprising:
 administering a soluble hyaluronidase enzyme to a subject at least once a week for a predetermined number of weeks in an amount sufficient to maintain a pharmacologically active level of the hyaluronidase in the plasma of at least about or 3 U/mL to prevent resynthesis of the substrate, hyaluronan, to levels prior to treatment, wherein:   the hyaluronidase enzyme is modified by conjugation to a polymer; and   the predetermined number of weeks is more than one week.   
     
     
         15 . The method of  claim 14 , wherein the predetermined number of weeks is at least two weeks. 
     
     
         16 . The method of  claim 14 , wheren the predetermined number of weeks is at least four weeks. 
     
     
         17 . The method of  claim 14 , wherein after the predetermined number of weeks, administration is discontinued for a first predetermined period of time, and then resumed for at least one week. 
     
     
         18 . The method of claim  claim 17 , wherein the cycle of administration and discontinuation of adminstration is repeated a plurality of times. 
     
     
         19 . The method of  claim 14 , wherein the hyaluronidase is administered in an amount sufficient to maintain a pharmacologically active level of the hyaluronidase in the plasma of at least or about 3 U/mL-12 U/mL. 
     
     
         20 . The method of  claim 14 , wherein the hyaluronidase is administered twice a week. 
     
     
         21 . The method of  claim 14 , wherein a single dose of hyaluronidase is 0.05 mg/kg-0.8 mg/kg. 
     
     
         22 . The method of  claim 14 , wherein:
 hyaluronan expression in a sample from the subject is measured prior to treatment; and   if the hyaluronan is elevated and at a level indicative of the disease or condition, the soluble hyaluronidase enzyme is administered to treat a disease or condition in which hyaluronan accumulates.   
     
     
         23 . The method of  claim 22 , wherein the sample from the subject is a tissue or body fluid. 
     
     
         24 . The method of  claim 23 , wherein the sample from the subject is a blood sample, tumor biopsy, cerebral spinal fluid, urine, sweat, semen or saliva sample. 
     
     
         25 . The method of  claim 14 , wherein the disease or condition is a disease or condition associated with elevated interstitial fluid pressure, decreased vascular volume, or increased water content in a tissue. 
     
     
         26 . The method of  claim 14 , wherein the hyaluronidase is a polymer-conjugated soluble PH20 hyaluronidase. 
     
     
         27 . The method of  claim 25 , wherein the the disease or condition is a cancer, disc pressure, or edema. 
     
     
         28 . The method of  claim 27 , wherein the disease or condition is edema, and the edema is caused by organ transplant, stroke or brain trauma. 
     
     
         29 . The method of  claim 14 , wherein the disease or condition is cancer, and the cancer is a tumor that has increased cellular and/or stromal expression of a hyaluronan, compared to a non-cancerous tissue of the same tissue type or compared to a non-metastatic tumor of the same tumor-type. 
     
     
         30 . The method of  claim 29 , wherein:
 the disease or condition is selected from among any one or more of ovarian cancer, in situ carcinoma (ISC), squamous cell carcinoma (SCC), prostate cancer, non-small cell lung cancer, breast cancer, brain cancer and colon cancer.   
     
     
         31 . The method of  claim 26 , wherein the soluble form of PH20 is a truncated human PH20 lacking all or a portion of a C-terminal GPI anchor. 
     
     
         32 . The method of  claim 26 , wherein the polymer-conjugated soluble PH20 hyaluronidase consists of the polymer and a polypeptide consisting of an amino acid sequence having at least 98% amino acid sequence identity with the amino acid sequence of SEQ ID NO:48, wherein the soluble PH20 hyaluronidase is N-glycosylated and neutral active. 
     
     
         33 . The method of  claim 26 , wherein the polymer-conjugated soluble PH20 hyaluronidase consists of the amino acid sequence set forth as amino acids 36-467, 36-468, 36-469, 36-470, 36-471, 36-472, 36-473, 36-474, 36-475, 36-476, 36-477, 36-478, 36-479, 36-480, 36-481, 36-482, or 36-483 of the amino acid sequence of SEQ ID NO:1, or an amino acid sequence having at least 95% amino acid sequence identity with the amino acid sequence set forth as amino acids 36-467, 36-468, 36-469, 36-470, 36-471, 36-472, 36-473, 36-474, 36-475, 36-476, 36-477, 36-478, 36-479, 36-480, 36-481, 36-482, or 36-483 of the amino acid sequence of SEQ ID NO:1. 
     
     
         34 . The method of  claim 14 , wherein:
 the soluble hyaluronidase is a PEGylated soluble PH20 hyaluronidase; and   the soluble PH20 hyaluronidase is selected from among a polypeptide consisting of the amino acid sequence set forth in SEQ ID NOs: 4-9 and 47-48, or variants thereof that have at least 98% amino acid sequence identity to the amino acid sequence of SEQ ID NO:48.   
     
     
         35 . The method of  claim 14 , wherein:
 the soluble hyaluronidase is PEGylated; and   PEGylation results from reaction with a PEG reagent selected from methoxy-poly(ethylene glycol)-succinimidyl butanoate (mPEG-SBA) (5 kDa); methoxy-poly(ethylene glycol)-succinimidyl butanoate (mPEG-SBA) (20 kDa); methoxy-poly(ethylene glycol)-succinimidyl butanoate (mPEG-SBA) (30 kDa); methoxy-poly(ethylene glycol)-succinimidyl a-methylbutanoate (mPEG-SMB) (20 kDa);   methoxy-poly(ethylene glycol)-succinimidyl a-methylbutanoate (mPEG-SMB) (30 kDa); methoxy-poly(ethylene glycol)-butyraldehyde (mPEG-butyraldehyde) (30 kDa), methoxy-poly(ethylene glycol)-succinimidyl propionate (mPEG-SPA) (20 kDa); methoxy-poly(ethylene glycol)-succinimidyl propionate (mPEG-SPA) (30 kDa); (methoxy-poly(ethylene glycol)) 2-N-hydroxysuccinimide ester (mPEG2-NHS) (10 kDa branched); (methoxy-poly(ethylene glycol)) 2-N-hydroxysuccinimide ester (mPEG2-NHS) (20 kDa branched); (methoxy-poly(ethylene glycol)) 2-N-hydroxysuccinimide ester (mPEG2-NHS) (40 kDa branched); (methoxy-poly(ethylene glycol)) 2-N-hydroxysuccinimide ester (mPEG2-NHS) (60 kDa branched); biotin-poly(ethylene glycol)-N-hydroxysuccinimide ester (biotin-PEG-NHS) (5 kDa biotinylated); poly(ethylene glycol)-p-nitrophenyl carbonate (PEG-p-nitrophenyl-carbonate) (30 kDa); and poly(ethylene glycol)- propionaldehyde (PEG-propionaldehyde) (30 kDa).   
     
     
         36 . The method of  claim 14 , wherein:
 the polymer-conjugated soluble hyaluronidase is PEGylated; and   the PEG is a branched or linear PEG.   
     
     
         37 . The method of  claim 35 , wherein:
 the polymer-conjugated soluble hyaluronidase is PEGylated; and   the PEG is a methoxy-PEG (mPEG), or is a linear N-hydroxysuccinimidyl ester of methoxy poly(ethylene glycol) butanoic acid.