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 disease or condition in which a hyaluronidase substrate accumulates, comprising:
 administering a soluble hyaluronidase enzyme to a subject more than 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 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.   
     
     
         2 . The method of  claim 1 , wherein the predetermined number of weeks is at least two weeks. 
     
     
         3 . The method of  claim 1 , wherein the predetermined number of weeks is two weeks, three weeks, or four weeks. 
     
     
         4 . The method of  claim 1 , 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. 
     
     
         5 . The method of  claim 4 , wherein the first predetermined period of time is at least one week. 
     
     
         6 . The method of  claim 4 , wherein the first predetermined period of time is one week, two weeks, three weeks or four weeks. 
     
     
         7 . The method of  claim 4 , wherein, after the first predetermined period of time, further administering the soluble hyaluronidase to the subject more than 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 to levels prior to treatment. 
     
     
         8 . The method of claim  claim 4 , wherein the cycle of administration and discontinuation of administration is repeated a plurality of times. 
     
     
         9 . The method of  claim 1 , wherein the hyaluronidase is administered in an amount sufficient to maintain a pharmacologically active level of the hyaluronidae in the plasma of at least or about 3 U/mL-12 U/mL. 
     
     
         10 . The method of  claim 1 , wherein the hyaluronidase is administered in an amount sufficient to maintain a pharmacologically active level of the hyaluronidase in the plasma of at least at or about 5 U/mL, 6 U/mL, 7 U/mL, 8 U/mL, 9 U/mL, 10 U/mL, 15 U/mL, 20 U/mL, 30 U/mL, 40 U/mL, 45 U/mL, 50 U/mL or more. 
     
     
         11 . The method of  claim 1 , 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 10 U/mL. 
     
     
         12 . The method of  claim 1 , wherein the hyalurondiase is administered twice a week. 
     
     
         13 . The method of  claim 1 , wherein the hyaluronidase is administered in an amount selected from among 0.02 mg/kg (of the subject), 0.03 mg/kg, 0.04 mg/kg, 0.05 mg/kg, 0.06 mg/kg, 0.07 mg/kg, 0.08 mg/kg, 0.09 mg/kg, 0.1 mg/kg, 0.15 mg/kg, 0.2 mg/kg, 0.25 mg/kg, 0.30 mg/kg, 0.35 mg/kg, 0.40 mg/kg, 0.45 mg/kg, 0.5 mg/kg, 0.55 mg.kg, 0.6 mg/kg, 0.7 mg/kg, 0.8 mg/kg, 0.9 mg/kg, 1.0 mg/kg, 1.1 mg/kg, 1.2 mg/kg, 1.3 mg/kg, 1.4 mg/kg, 1.5 mg/kg or more. 
     
     
         14 . The method of  claim 1 , wherein the hyaluronidase is administered in an amount selected from among 0.05 mg/kg-0.8 mg/kg. 
     
     
         15 . The method of  claim 1 , wherein the hyalurondiase is administered in an amount that is or is about 50,000 Units (U); 60,000 U; 70,000 U; 80,000 U; 90,000 U; 100,000 U; 200,000 U; 300,000 U; 400,000 U; 500,000 U; 600,000 U; 700,000 U; 800,000 U; 900,000 U; 1,000,000 U; 1,500,000 U; 2,000,000 U; 2,500,000 U; 3,000,000 U; 3,500,000 U; 4,000,000 U or more. 
     
     
         16 . The method of  claim 1 , wherein the hyaluronidase substrate is hyaluronan. 
     
     
         17 . The method of  claim 1 , wherein hyaluronan expression in a sample from the subject is measured prior to treatment. 
     
     
         18 . A method for treating a hyaluronan-associated disease or condition in a subject, comprising:
 (a) measuring hyaluronan expression or hyaluron in a sample from the subject; and   (b) if the hyaluronan expression or hyaluron in the sample from the subject is elevated or at a level indicative of the disease or condition, administering a composition containing a soluble hyaluronidase to the subject, wherein the soluble hyaluronidase is modified by conjugation to a polymer.   
     
     
         19 . The method of  claim 18 , wherein the sample from the subject is a tissue or body fluid. 
     
     
         20 . The method of  claim 18 , wherein the sample from the subject is a blood sample, tumor biopsy, cerebral spinal fluid, urine, sweat, semen or saliva sample. 
     
     
         21 . The method of  claim 1 , further comprising: administering a second and different agent for treating the disease or condition to the subject. 
     
     
         22 . The method of  claim 21 , wherein the second agent and the composition containing the soluble hyaluronidase are administered in a single composition. 
     
     
         23 . The method of  claim 21 , wherein the second agent and the composition containing the soluble hyalurondiase are administered separately. 
     
     
         24 . The method of  claim 23 , wherein the second agent and the composition containing the soluble hyaluronidase are administered simultaneously, sequentially or intermittently in any order. 
     
     
         25 . The method of  claim 24 , wherein the second agent is administered after administration of the composition containing the soluble hyaluronidase. 
     
     
         26 . The method of  claim 25 , wherein the second agent is administered after the first administration of the soluble hyaluronidase in the cycle of administration, and optionally after one or more subsequent administrations of the soluble hyaluronidase. 
     
     
         27 . The method of  claim 26 , wherein the second agent is administered after each subsequent administration of the soluble hyaluronidase. 
     
     
         28 . The method of  claim 26 , wherein the second agent is administered after every other subsequent administration of the soluble hyaluronidase. 
     
     
         29 . The method of  claim 26 , wherein the second agent is administered once a week, once every two weeks, once every three weeks or once a month. 
     
     
         30 . The method of  claim 25 , wherein the second agent is administered at least 0.5 minutes, at least one minute, at least five minutes, at least fifteen minutes, at least thirty minutes, at least one hour or more than one hour after the composition containing the soluble hyaluronidase is administered. 
     
     
         31 . The method of  claim 25 , wherein the second agent is administered at least or two hours, four hours, six hours, eight hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours or 24 hours after the composition containing the soluble hyaluronidase is administered. 
     
     
         32 . The method of of  claim 25 , wherein the second agent is administered at least forty-eight hours after the composition containing the soluble hyaluronidase. 
     
     
         33 . The method of of  claim 25 , wherein second agent is administered at least seventy-two hours after the composition containing the soluble hyaluronidase. 
     
     
         34 . The method of  claim 1 , wherein the polymer is a sialation or pegylation moiety. 
     
     
         35 . The method of  claim 21 , wherein the second agent is an anti-cancer agent or treatment. 
     
     
         36 . The method of  claim 35 , wherein the anti-cancer agent or treatment is selected from among a chemotherapeutic agent, radiation therapy an antibody, a peptide, a gene therapy vector, a virus and a nucleic acid. 
     
     
         37 . The method of  claim 21 , wherein:
 the second agent is an anti-cancer agent selected from among Acivicins; Aclarubicins; Acodazoles; Acronines; Adozelesins; Aldesleukins; Alemtuzumabs; Alitretinoins (9-Cis-Retinoic Acids); Allopurinols; Altretamines; Alvocidibs; Ambazones; Ambomycins; Ametantrones; Amifostines; Aminoglutethimides; Amsacrines; Anastrozoles; Anaxirones; Ancitabines; Anthramycins; Apaziquones; Argimesnas; Arsenic Trioxides; Asparaginases; Asperlins; Atrimustines; Azacitidines; Azetepas; Azotomycins; Banoxantrones; Batabulins; Batimastats; BCG Live; Benaxibines; Bendamustines; Benzodepas; Bexarotenes; Bevacizumab; Bicalutamides; Bietaserpines; Biricodars; Bisantrenes; Bisantrenes; Bisnafide Dimesylates; Bizelesins; Bleomycins; Bortezomibs; Brequinars; Bropirimines; Budotitanes; Busulfans; Cactinomycins; Calusterones; Canertinibs; Capecitabines; Caracemides; Carbetimers; Carboplatins; Carboquones; Carmofurs; Carmustines with Polifeprosans; Carmustines; Carubicins; Carzelesins; Cedefingols; Celecoxibs; Cemadotins; Chlorambucils; Cioteronels; Cirolemycins; Cisplatins; Cladribines; Clanfenurs; Clofarabines; Crisnatols; Cyclophosphamides; Cytarabine liposomals; Cytarabines; Dacarbazines; Dactinomycins; Darbepoetin Alfas; Daunorubicin liposomals; Daunorubicins/Daunomycins; Daunorubicins; Decitabines; Denileukin Diftitoxes; Dexniguldipines; Dexonnaplatins; Dexrazoxanes; Dezaguanines; Diaziquones; Dibrospidiums; Dienogests; Dinalins; Disermolides; Docetaxels; Dofequidars; Doxifluridines; Doxorubicin liposomals; Doxorubicin HCL; Docorubicin HCL liposome injection; Doxorubicins; Droloxifenes; Dromostanolone Propionates; Duazomycins; Ecomustines; Edatrexates; Edotecarins; Eflomithines; Elacridars; Elinafides; Elliott's B Solutions; Elsamitrucins; Emitefurs; Enloplatins; Enpromates; Enzastaurins; Epipropidines; Epirubicins; Epoetin alfas; Eptaloprosts; Erbulozoles; Esorubicins; Estramustines; Etanidazoles; Etoglucids; Etoposide phosphates; Etoposide VP-16s; Etoposides; Etoprines; Exemestanes; Exisulinds; Fadrozoles; Fazarabines; Fenretinides; Filgrastims; Floxuridines; Fludarabines; Fluorouracils; 5-fluorouracils; Fluoxymesterones; Flurocitabines; Fosquidones; Fostriecins; Fostriecins; Fotretamines; Fulvestrants; Galarubicins; Galocitabines; Gemcitabines; Gemtuzumabs/Ozogamicins; Geroquinols; Gimatecans; Gimeracils; Gloxazones; Glufosfamides; Goserelin acetates; Hydroxyureas; Ibritumomabs/Tiuxetans; Idarubicins; Ifosfamides; Ilmofosines; Ilomastats; Imatinib mesylates; Imexons; Improsulfans; Indisulams; Inproquones; Interferon alfa-2as; Interferon alfa-2bs; Interferon Alfas; Interferon Betas; Interferon Gammas; Interferons; Interleukin-2s and other Interleukins (including recombinant Interleukins); Intoplicines; Iobenguanes [131-I]; Iproplatins; Irinotecans; Irsogladines; Ixabepilones; Ketotrexates; L-Alanosines; Lanreotides; Lapatinibs; Ledoxantrones; Letrozoles; Leucovorins; Leuprolides; Leuprorelins (Leuprorelides); Levamisoles; Lexacalcitols; Liarozoles; Lobaplatins; Lometrexols; Lomustines/CCNUs; Lomustines; Lonafamibs; Losoxantrones; Lurtotecans; Mafosfamides; Mannosulfans; Marimastats; Masoprocols; Maytansines; Mechlorethamines; Meclorethamines/Nitrogen mustards; Megestrol acetates; Megestrols; Melengestrols; Melphalans; MelphalanslL-PAMs; Menogarils; Mepitiostanes; Mercaptopurines; 6-Mecaptopurine; Mesnas; Metesinds; Methotrexates; Methoxsalens; Metomidates; Metoprines; Meturedepas; Miboplatins; Miproxifenes; Misonidazoles; Mitindomides; Mitocarcins; Mitocromins; Mitoflaxones; Mitogillins; Mitoguazones; Mitomalcins; Mitomycin Cs; Mitomycins; Mitonafides; Mitoquidones; Mitospers; Mitotanes; Mitoxantrones; Mitozolomides; Mivobulins; Mizoribines; Mofarotenes; Mopidamols; Mubritinibs; Mycophenolic Acids; Nandrolone Phenpropionates; Nedaplatins; Nelzarabines; Nemorubicins; Nitracrines; Nocodazoles; Nofetumomabs; Nogalamycins; Nolatrexeds; Nortopixantrones; Octreotides; Oprelvekins; Ormaplatins; Ortataxels; Oteracils; Oxaliplatins; Oxisurans; Oxophenarsines; Paclitaxels; Pamidronates; Patubilones; Pegademases; Pegaspargases; Pegfilgrastims; Peldesines; Peliomycins; Pelitrexols; Pemetrexeds; Pentamustines; Pentostatins; Peplomycins; Perfosfamides; Perifosines; Picoplatins; Pinafides; Pipobromans; Piposulfans; Pirfenidones; Piroxantrones; Pixantrones; Plevitrexeds; Plicamycid Mithramycins; Plicamycins; Plomestanes; Plomestanes; Porfimer sodiums; Porfimers; Porfiromycins; Prednimustines; Procarbazines; Propamidines; Prospidiums; Pumitepas; Puromycins; Pyrazofurins; Quinacrines; Ranimustines; Rasburicases; Riboprines; Ritrosulfans; Rituximabs; Rogletimides; Roquinimexs; Rufocromomycins; Sabarubicins; Safingols; Sargramostims; Satraplatins; Sebriplatins; Semustines; Simtrazenes; Sizofirans; Sobuzoxanes; Sorafenibs; Sparfosates; Sparfosic Acids; Sparsomycins; Spirogermaniums; Spiromustines; Spiroplatins; Spiroplatins; Squalamines; Streptonigrins; Streptovarycins; Streptozocins; Sufosfamides; Sulofenurs; Sunitinib Malate; 6-thioguanine (6-TG); Tacedinalines; Talcs; Talisomycins; Tallimustines; Tamoxifens; Tariquidars; Tauromustines; Tecogalans; Tegafurs; Teloxantrones; Temoporfins; Temozolomides; Teniposides/VM-26s; Teniposides; Teroxirones; Testolactones; Thiamiprines; Thioguanines; Thiotepas; Tiamiprines; Tiazofurins; Tilomisoles; Tilorones; Timcodars; Timonacics; Tirapazamines; Topixantrones; Topotecans; Toremifenes; Tositumomabs; Trabectedins (Ecteinascidin 743); Trastuzumabs; Trestolones; Tretinoins/ATRA; Triciribines; Trilostanes; Trimetrexates; Triplatin Tetranitrates; Triptorelins; Trofosfamides; Tubulozoles; Ubenimexs; Uracil Mustards; Uredepas; Valrubicins; Valspodars; Vapreotides; Verteporfins; Vinblastines; Vincristines; Vindesines; Vinepidines; Vinflunines; Vinformides; Vinglycinates; Vinleucinols; Vinleurosines; Vinorelbines; Vinrosidines; Vintriptols; Vinzolidines; Vorozoles; Xanthomycin As (Guamecyclines); Zeniplatins; Zilascorbs [2-H]; Zinostatins; Zoledronate; Zorubicins; and Zosuquidars.   
     
     
         38 . The method of  claim 21 , wherein the soluble hyaluronidase and/or second agent is administered locally or systemically. 
     
     
         39 . The method of  claim 38 , wherein the soluble hyaluronidase and/or second agent is administered orally, intravenously (IV), subcutaneously, intramuscularly, intra-tumorally, radermally, topically, transdermally, rectally or sub-epidermally. 
     
     
         40 . The method of  claim 39 , wherein the soluble hyaluronidase and/or second agent is administered intravenously. 
     
     
         41 . The method of  claim 39 , wherein the soluble hyaluronidase and/or second agent is administered intra-tumorally. 
     
     
         42 . The method of  claim 1 , further comprising comparing the expression of hyaluronan in the sample from the subject to expression in a control sample or a standard. 
     
     
         43 . The method of  claim 1 , wherein the disease or condition in which a hyaluronidase substrate accumulates is associated with high interstitial fluid pressure. 
     
     
         44 . The method of  claim 1 , wherein the disease or condition is selected from among disc pressure, cancer and edema. 
     
     
         45 . The method of  claim 44 , wherein the disease or condition is edema, and the edema is caused by organ transplant, stroke or brain trauma. 
     
     
         46 . The method of  claim 44 , wherein the disease or condition is cancer, and the cancer is a tumor. 
     
     
         47  The method of  claim 46 , wherein the disease or condition is a solid tumor. 
     
     
         48 . The method of  claim 47 , wherein the tumor 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. 
     
     
         49 . The method of  claim 46 , wherein:
 the disease or condition is selected from among any one or more of a late-stage cancer, a metastatic cancer and an undifferentiated cancer.   
     
     
         50 . The method of  claim 1 , 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, pancreatic cancer, non-small cell lung cancer, breast cancer, brain cancer and colon cancer.   
     
     
         51 . The method of  claim 1 , wherein:
 the soluble hyaluronidase is a soluble PH20.   
     
     
         52 . The method of  claim 51 , wherein the PH20 is selected from among an ovine, mouse, monkey, bovine, bacterial and human PH20. 
     
     
         53 . The method of  claim 51 , wherein the soluble form of PH20 is a soluble PH20 that has been truncated to remove a C-terminal GPI. 
     
     
         54 . The method of  claim 1 , wherein the soluble hyaluronidase has a sequence of amino acids included in SEQ ID NO:1 or a sequence that has at least about 91% amino acid sequence identity with a sequence of amino acids included in SEQ ID NO: 1, whereby the soluble hyaluronidase is soluble, N-glycosylated and neutral active. 
     
     
         55 . The method of  claim 1 , wherein the soluble hyaluronidase includes a sequence of amino acids set forth in SEQ ID NO: 1 that is truncated at an amino acid residue that is or is between amino acid residues 467 to 483. 
     
     
         56 . The method of  claim 1 , wherein the soluble hyaluronidase includes a sequence of amino acids set forth in SEQ ID NO: 1 that is truncated at an amino acid residue selected from among 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482 and 483. 
     
     
         57 . The method of  claim 1 , wherein the soluble hyaluronidase is secreted in CHO cells. 
     
     
         58 . The method of  claim 1 , wherein the soluble hyaluronidase has the sequence of amino acids 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 SEQ ID NO: 1, or has at least about 91% amino acid sequence identity with a sequence of amino acids 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 SEQ ID NO:1. 
     
     
         59 . The method of  claim 1 , wherein the soluble hyaluronidase is selected from among polypeptides containing a sequence of amino acids set forth in any of SEQ ID NOS: 4-9 and 46-48, and allelic variants, species variants and other variants thereof. 
     
     
         60 . The method of  claim 1 , wherein the soluble hyaluronidase is a polypeptide encoded by a sequence of nucleic acids that encodes a sequence of amino acids set forth in SEQ ID NO:4. 
     
     
         61 . The method of  claim 1 , wherein the soluble hyaluronidase is selected from among polpeptides encoded by a sequence of nucleic acids that encodes a sequence of amino acids set forth in any of SEQ ID NOS: 4-9. 
     
     
         62 . The method of  claim 61 , wherein the soluble hyaluronidase is produced by expression in CHO cells. 
     
     
         63 . The method of  claim 62 , wherein the soluble hyaluronidase is designated rHuPH20. 
     
     
         64 . The method of  claim 1 , wherein the soluble hyaluronidase is glycosylated. 
     
     
         65 . The method of  claim 1 , wherein the polymer conjugated to the soluble hyaluronidase comprises a pegylation moiety (PEG). 
     
     
         66 . The method of  claim 65 , wherein the polymer is selected from among 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 (mPEG 2 -NHS) (10 kDa branched); (methoxy-poly(ethylene glycol)) 2 -N-hydroxysuccinimide ester (mPEG 2 -NHS) (20 kDa branched); (methoxy-poly(ethylene glycol)) 2 -N-hydroxysuccinimide ester (mPEG 2 -NHS) (40 kDa branched); (methoxy-poly(ethylene glycol)) 2 -N-hydroxysuccinimide ester (mPEG 2 -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)-priopionaldehyde (PEG-propionaldehyde) (30 kDa). 
     
     
         67 . The method of  claim 66 , wherein the PEG is a branched or linear PEG. 
     
     
         68 . The method of  claim 67 , wherein the PEG is a linear PEG. 
     
     
         69 . The method of  claim 65 , wherein the PEG is a methoxy-PEG (mPEG). 
     
     
         70 . The method of  claim 69 , wherein the PEG is a linear N-hydroxysuccinimidyl ester of methoxy poly(ethylene glycol) butanoic acid. 
     
     
         71 . The method of  claim 69 , wherein the PEG has a molecular weight of 30 or about 30 kilodaltons. 
     
     
         72 . The method of  claim 1 , wherein the treatment effects a reduction in the size of a tumor in the subject. 
     
     
         73 . The method of  claim 18 , further comprising:
 administering a second and different agent for treating the disease or condition to the subject.   
     
     
         74 . The method of  claim 18 , wherein:
 the soluble hyaluronidase is a soluble PH20.   
     
     
         75 . A composition, comprising a soluble hyaluronidase in an amount sufficient for maintaining a plasma level of the hyaluronidase enzyme in plasma at a level of at least 3 U/mL for at least a week, wherein the soluble hyaluronidase is conjugated to a polymer. 
     
     
         76 . The composition of  claim 75 , wherein the composition is for administration at least twice a week for more than at least one week. 
     
     
         77 . The composition of  claim 75 , wherein:
 the composition contains at least or about 2.0 mg-60 mg of the soluble hyaluronidase conjugated to a polymer; and   the soluble hyaluronidase conjugated to a polymer has a specific activity of at least or about 20,000 U/mg, 25,000 U/mg, 30,000 U/mg, 31,000 U/mg, 32,000 U/mg, 33,000 U/mg, 34,000 U/mg, 35,000 U/mg, 36,000 U/mg, 37,000 U/mg, 38, 000 U/mg, 39,000 U/mg, 40,000 U/mg, 45,000 U/mg, 50,000 U/mg, 55,000 U/mg, 60,000 U/mg or more.   
     
     
         78 . The composition of  claim 75 , wherein the composition is at least 10 mL/per administration. 
     
     
         79 . The composition of  claim 75 , wherein the composition is formulated for administration orally, intravenously (IV), subcutaneously, intramuscularly, intra-tumorally, radermally, topically, transdermally, rectally or sub-epidermally. 
     
     
         80 . The composition of  claim 75 , wherein the composition is formulated for intravenous administration. 
     
     
         81 . The composition of  claim 75 , further comprising histidine and/or NaCl. 
     
     
         82 . The composition of  claim 81 , wherein the composition is formulated with at or about 10 mM histidine and/or 130 mM NaCl. 
     
     
         83 . The composition of  claim 75 , wherein the composition has a pH that is or is about 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1 or 7.2. 
     
     
         84 . The composition of  claim 83  that has a pH of or about 6.5. 
     
     
         85 . The composition of  claim 75 , wherein the level of hyaluronidase in the plasma is at least or about 3 U/mL-12 U/mL. 
     
     
         86 . The composition of  claim 75 , wherein the polymer is a sialation or pegylation moiety. 
     
     
         87 . A combination, comprising:
 a composition containing a soluble hyaluronidase in an amount sufficient for maintaining a plasma level of the hyaluronidase enzyme in plasma at a level of at least 3 U/mL for at least a week, wherein the soluble hyaluronidase is conjugated to a polymer; and   a second composition containing an agent for treating a hyaluronan-associated disease or condition.   
     
     
         88 . The combination of  claim 87 , wherein the composition containing a soluble hyaluronidase is for administration at least twice a week for more than at least one week. 
     
     
         89 . The combination of  claim 87 , wherein:
 the composition containing a soluble hyaluronidase contains at least or about 2.0 mg-60 mg of the soluble hyaluronidase conjugated to a polymer; and   the soluble hyaluronidase conjugated to a polymer has a specific activity of at least or about 20,000 U/mg, 25,000 U/mg, 30,000 U/mg, 31,000 U/mg, 32,000 U/mg, 33,000 U/mg, 34,000 U/mg, 35,000 U/mg, 36,000 U/mg, 37,000 U/mg, 38,000 U/mg, 39,000 U/mg, 40,000 U/mg, 45,000 U/mg, 50,000 U/mg, 55,000 U/mg, 60,000 U/mg or more.   
     
     
         90 . The combination of  claim 87 , wherein the first and second compositions are co-formulated or provided separately. 
     
     
         91 . The combination of  claim 87 , wherein the polymer is a sialation or pegylation moiety. 
     
     
         92 . The combination of of  claim 87 , wherein the second agent is an anti-cancer agent or treatment. 
     
     
         93 . The combination of  claim 92 , wherein the anti-cancer agent or treatment is selected from among a chemotherapeutic agent, radiation therapy an antibody, a peptide, a gene therapy vector, a virus and a nucleic acid. 
     
     
         94 . The combination of  claim 87 , wherein:
 the second agent is an anti-cancer agent selected from among Acivicins; Aclarubicins; Acodazoles; Acronines; Adozelesins; Aldesleukins; Alemtuzumabs; Alitretinoins (9-Cis-Retinoic Acids); Allopurinols; Altretamines; Alvocidibs; Ambazones; Ambomycins; Ametantrones; Amifostines; Aminoglutethimides; Amsacrines; Anastrozoles; Anaxirones; Ancitabines; Anthramycins; Apaziquones; Argimesnas; Arsenic Trioxides; Asparaginases; Asperlins; Atrimustines; Azacitidines; Azetepas; Azotomycins; Banoxantrones; Batabulins; Batimastats; BCG Live; Benaxibines; Bendamustines; Benzodepas; Bexarotenes; Bevacizumab; Bicalutamides; Bietaserpines; Biricodars; Bisantrenes; Bisantrenes; Bisnafide Dimesylates; Bizelesins; Bleomycins; Bortezomibs; Brequinars; Bropirimines; Budotitanes; Busulfans; Cactinomycins; Calusterones; Canertinibs; Capecitabines; Caracemides; Carbetimers; Carboplatins; Carboquones; Carmofurs; Carmustines with Polifeprosans; Carmustines; Carubicins; Carzelesins; Cedefingols; Celecoxibs; Cemadotins; Chlorambucils; Cioteronels; Cirolemycins; Cisplatins; Cladribines; Clanfenurs; Clofarabines; Crisnatols; Cyclophosphamides; Cytarabine liposomals; Cytarabines; Dacarbazines; Dactinomycins; Darbepoetin Alfas; Daunorubicin liposomals; Daunorubicins/Daunomycins; Daunorubicins; Decitabines; Denileukin Diftitoxes; Dexniguldipines; Dexonnaplatins; Dexrazoxanes; Dezaguanines; Diaziquones; Dibrospidiums; Dienogests; Dinalins; Disermolides; Docetaxels; Dofequidars; Doxifluridines; Doxorubicin liposomals; Doxorubicin HCL; Docorubicin HCL liposome injection; Doxorubicins; Droloxifenes; Dromostanolone Propionates; Duazomycins; Ecomustines; Edatrexates; Edotecarins; Eflornithines; Elacridars; Elinafides; Elliott's B Solutions; Elsamitrucins; Emitefurs; Enloplatins; Enpromates; Enzastaurins; Epipropidines; Epirubicins; Epoetin alfas; Eptaloprosts; Erbulozoles; Esorubicins; Estramustines; Etanidazoles; Etoglucids; Etoposide phosphates; Etoposide VP-16s; Etoposides; Etoprines; Exemestanes; Exisulinds; Fadrozoles; Fazarabines; Fenretinides; Filgrastims; Floxuridines; Fludarabines; Fluorouracils; 5-fluorouracils; Fluoxymesterones; Flurocitabines; Fosquidones; Fostriecins; Fostriecins; Fotretamines; Fulvestrants; Galarubicins; Galocitabines; Gemcitabines; Gemtuzumabs/Ozogamicins; Geroquinols; Gimatecans; Gimeracils; Gloxazones; Glufosfamides; Goserelin acetates; Hydroxyureas; Ibritumomabs/Tiuxetans; Idarubicins; Ifosfamides; Ilmofosines; Ilomastats; Imatinib mesylates; Imexons; Improsulfans; Indisulams; Inproquones; Interferon alfa-2as; Interferon alfa-2bs; Interferon Alfas; Interferon Betas; Interferon Gammas; Interferons; Interleukin-2s and other Interleukins (including recombinant Interleukins); Intoplicines; Iobenguanes [131-I]; Iproplatins; Irinotecans; Irsogladines; Ixabepilones; Ketotrexates; L-Alanosines; Lanreotides; Lapatinibs; Ledoxantrones; Letrozoles; Leucovorins; Leuprolides; Leuprorelins (Leuprorelides); Levamisoles; Lexacalcitols; Liarozoles; Lobaplatins; Lometrexols; Lomustines/CCNUs; Lomustines; Lonafarnibs; Losoxantrones; Lurtotecans; Mafosfamides; Mannosulfans; Marimastats; Masoprocols; Maytansines; Mechlorethamines; Meclorethamines/Nitrogen mustards; Megestrol acetates; Megestrols; Melengestrols; Melphalans; MelphalanslL-PAMs; Menogarils; Mepitiostanes; Mercaptopurines; 6-Mecaptopurine; Mesnas; Metesinds; Methotrexates; Methoxsalens; Metomidates; Metoprines; Meturedepas; Miboplatins; Miproxifenes; Misonidazoles; Mitindomides; Mitocarcins; Mitocromins; Mitoflaxones; Mitogillins; Mitoguazones; Mitomalcins; Mitomycin Cs; Mitomycins; Mitonafides; Mitoquidones; Mitospers; Mitotanes; Mitoxantrones; Mitozolomides; Mivobulins; Mizoribines; Mofarotenes; Mopidamols; Mubritinibs; Mycophenolic Acids; Nandrolone Phenpropionates; Nedaplatins; Nelzarabines; Nemorubicins; Nitracrines; Nocodazoles; Nofetumomabs; Nogalamycins; Nolatrexeds; Nortopixantrones; Octreotides; Oprelvekins; Ormaplatins; Ortataxels; Oteracils; Oxaliplatins; Oxisurans; Oxophenarsines; Paclitaxels; Pamidronates; Patubilones; Pegademases; Pegaspargases; Pegfilgrastims; Peldesines; Peliomycins; Pelitrexols; Pemetrexeds; Pentamustines; Pentostatins; Peplomycins; Perfosfamides; Perifosines; Picoplatins; Pinafides; Pipobromans; Piposulfans; Pirfenidones; Piroxantrones; Pixantrones; Plevitrexeds; Plicamycid Mithramycins; Plicamycins; Plomestanes; Plomestanes; Porfimer sodiums; Porfimers; Porfiromycins; Prednimustines; Procarbazines; Propamidines; Prospidiums; Pumitepas; Puromycins; Pyrazofurins; Quinacrines; Ranimustines; Rasburicases; Riboprines; Ritrosulfans; Rituximabs; Rogletimides; Roquinimexs; Rufocromomycins; Sabarubicins; Safingols; Sargramostims; Satraplatins; Sebriplatins; Semustines; Simtrazenes; Sizofirans; Sobuzoxanes; Sorafenibs; Sparfosates; Sparfosic Acids; Sparsomycins; Spirogermaniums; Spiromustines; Spiroplatins; Spiroplatins; Squalamines; Streptonigrins; Streptovarycins; Streptozocins; Sufosfamides; Sulofenurs; Sunitinib Malate; 6-thioguanine (6-TG); Tacedinalines; Talcs; Talisomycins; Tallimustines; Tamoxifens; Tariquidars; Tauromustines; Tecogalans; Tegafurs; Teloxantrones; Temoporfins; Temozolomides; Teniposides/VM-26s; Teniposides; Teroxirones; Testolactones; Thiamiprines; Thioguanines; Thiotepas; Tiamiprines; Tiazofurins; Tilomisoles; Tilorones; Timcodars; Timonacics; Tirapazamines; Topixantrones; Topotecans; Toremifenes; Tositumomabs; Trabectedins (Ecteinascidin 743); Trastuzumabs; Trestolones; Tretinoins/ATRA; Triciribines; Trilostanes; Trimetrexates; Triplatin Tetranitrates; Triptorelins; Trofosfamides; Tubulozoles; Ubenimexs; Uracil Mustards; Uredepas; Valrubicins; Valspodars; Vapreotides; Verteporfins; Vinblastines; Vincristines; Vindesines; Vinepidines; Vinflunines; Vinformides; Vinglycinates; Vinleucinols; Vinleurosines; Vinorelbines; Vinrosidines; Vintriptols; Vinzolidines; Vorozoles; Xanthomycin As (Guamecyclines); Zeniplatins; Zilascorbs [2-H]; Zinostatins; Zoledronate; Zorubicins; and Zosuquidars.