Extended soluble PH20 polypeptides and uses thereof

• US 8,927,249 B2
• Filed: 12/09/2009
• Issued: 01/06/2015
• Est. Priority Date: 12/09/2008
• Status: Active Grant

• Current Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Original Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Inventors: Wei, Ge, Selvam, Krishnasamy Panneer, Bookbinder, Louis, Frost, Gregory I.
• Primary Examiner(s): Swope, Sheridan
• Application Number:
  US12/653,245
• Publication Number:
  US 20100143457A1
• Time in Patent Office: 1,854 Days
• Field of Search: None
• US Class Current: 435/200
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/6435 the peptide or protein in the drug conjugate being a connective tissue peptide, e.g. collagen, fibronectin or gelatin
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/08 of the prostate
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 33/04 Amoebicides
• CPC Class Codes: A61P 33/06 Antimalarials
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C12N 9/2408 : acting on alpha -1,4-glucosidic bonds
• CPC Class Codes: C12N 9/2474 : Hyaluronoglucosaminidase (3.2.1.35), i.e. hyaluronidase
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Soluble PH20 polypeptides are provided, including extended soluble PH20 polypeptides, and uses thereof. Also provided are other C-terminally truncated PH20 polypeptides and partially deglycosylated PH20 polypeptides and uses thereof.

Claims

1. An extended soluble PH20 hyaluronidase (esPH20), wherein:

the esPH20 polypeptide exhibits hyaluronidase activity at neutral pH;

the esPH20 polypeptide is secreted when expressed in a mammalian cell;

the esPH20 polypeptide is soluble; and

the esPH20 polypeptide is selected from among;

a polypeptide that consists of the sequence of amino acids set forth as amino acid residues 36-495, 36-496, 36-497, 36-498, 36-499 or 36-500 of SEQ ID NO;

107;

or

a polypeptide that contains amino acid substitutions in the sequence of amino acids set forth as amino acids 36-495, 36-496, 36-497, 36-498, 36-499 or 36-500 of SEQ ID NO;

107, whereby the amino acid-substituted polypeptide consists of a sequence of amino acids that has at least 95% sequence identity with the corresponding sequence of amino acids set forth as amino acids 36-495, 36-496, 36-497, 36-498, 36-499 or 36-500 of SEQ ID NO;

107.

2. The esPH20 of claim 1 that is N-glycosylated.

3. The esPH20 of claim 2, wherein the esPH20 is N-glycosylated and comprises at least an N-acetylglucosamine moiety linked to each of at least three asparagine (N) residues.

4. The esPH20 of claim 3, wherein the three asparagine residues correspond to amino acid residues 235, 368 and 393 of SEQ ID NO:

107.

5. The esPH20 of claim 3 that is modified by a polymer.

6. The esPH20 of claim 1 that consists of the sequence of amino acids set forth as amino acids 36-495, 36-496, 36-497, 36-498, 36-499 or 36-500 of SEQ ID NO:

107.

7. The esPH20 of claim 1 that is modified by a modification selected from among sialation, albumination, farnesylation, carboxylation, hydroxylation and phosphorylation.

8. The esPH20 of claim 1 that is modified by a polymer.

9. The esPH8 of claim 8, wherein the polymer is dextran or PEG.

10. The esPH20 of claim 1 that is not bifucosylated.

11. The esPH20 of claim 1 that is purified or isolated.

12. A conjugate, comprising the esPH20 of claim 1.

13. The conjugate of claim 12, wherein the esPH20 is conjugated to a moiety selected from among a multimerization domain, toxin, detectable label and drug.

14. The conjugate of claim 13, wherein the esPH20 is conjugated to an Fc domain.

15. The conjugate of claim 12, wherein the conjugated moiety is linked directly or via a linker to the C-terminus or N-terminus of the esPH20.

16. A composition, comprising the esPH20 polypeptide of claim 1.

17. A composition, comprising a plurality of esPH20 polypeptides of claim 1.

18. The composition of claim 16, wherein the esPH20 polypeptide is secreted from CHO cells.

19. The composition of claim 16 that is a pharmaceutical composition.

20. The composition of claim 19 comprising an additional therapeutic agent.

21. The composition of claim 20, wherein the therapeutic agent is selected from among a chemotherapeutic agent, an analgesic agent, an anti-inflammatory agent, an antimicrobial agent, an amoebicidal agent, a trichomonacidal agent, an anti-parkinson agent, an anti-malarial agent, an anticonvulsant agent, an anti-depressant agent, an antiarthritics agent, an anti-fungal agent, an antihypertensive agent, an antipyretic agent, an anti-parasite agent, an antihistamine agent, an alpha-adrenergic agonist agent, an alpha blocker agent, an anesthetic agent, a bronchial dilator agent, a biocide agent, a bactericide agent, a bacteriostat agent, a beta adrenergic blocker agent, a calcium channel blocker agent, a cardiovascular drug agent, a contraceptive agent, a decongestant agent, a diuretic agent, a depressant agent, a diagnostic agent, a electrolyte agent, a hypnotic agent, a hormone agent, a hyperglycemic agent, a muscle relaxant agent, a muscle contractant agent, an ophthalmic agent, a parasympathomimetic agent, a psychic energizer agent, a sedative agent, a sympathomimetic agent, a tranquilizer agent, an urinary agent, a vaginal agent, a viricide agent, a vitamin agent, a non-steroidal anti-inflammatory agent, an angiotensin converting enzyme inhibitor agent, a polypeptide, a protein, a nucleic acid, a drug, an organic molecule and a sleep inducer.

22. The composition of claim 20, wherein the therapeutic agent is selected from among an antibody, an immunoglobulin, a bisphosphonate, a cytokine, a chemotherapeutic agent and an insulin.

23. The composition of claim 22, wherein the insulin is a fast-acting insulin and the bisphosphonate is zoledronic acid.

24. The composition of claim 20, wherein the additional therapeutic agent is 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;

Dexonas;

Dexrazoxanes;

Dezaguanines;

Diaziquones;

Dibrospidiums;

Dienogests;

Dinalins;

Disermolides;

Docetaxels;

Dofequidars;

Doxifluridines;

Doxorubicin liposomals;

Doxorubicin HCL;

Doxorubicin 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;

Fluorocitabines;

Fosquidones;

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-2 as;

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 (Leuprolides);

Levamisoles;

Lexacalcitols;

Liarozoles;

Lobaplatins;

Lometrexols;

Lomustines/CCNUs;

Lomustines;

Lonafarnibs;

Losoxantrones;

Lurtotecans;

Mafosfamides;

Mannosulfans;

Marimastats;

Masoprocols;

Maytansines;

Mechlorethamines;

Mechlorethamines/Nitrogen mustards;

Megestrol acetates;

Megestrols;

Melengestrols;

Melphalans;

L-PAMs;

Menogarils;

Mepitiostanes;

Mercaptopurines;

6-Mercaptopurine;

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;

Nelarabines;

Nemorubicins;

Nitracrines;

Nocodazoles;

Nofetumomabs;

Nogalamycins;

Nolatrexeds;

Nortopixantrones;

Octreotides;

Oprelvekins;

Ormaplatins;

Ortataxels;

Oteracils;

Oxaliplatins;

Oxisurans;

Oxophenarsines;

Paclitaxels;

Pamidronates;

Patupilones;

Pegademases;

Pegaspargases;

Pegfilgrastims;

Peldesines;

Peliomycins;

Pelitrexols;

Pemetrexeds;

Pentamustines;

Pentostatins;

Peplomycins;

Perfosfamides;

Perifosines;

Picoplatins;

Pinafides;

Pipobromans;

Piposulfans;

Pirfenidones;

Piroxantrones;

Pixantrones;

Plevitrexeds;

Plicamycin 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;

Sizofurans;

Sobuzoxanes;

Sorafenibs;

Sparfosates;

Sparfosic Acids;

Sparsomycins;

Spirogermaniums;

Spiromustines;

Spiroplatins;

Spiroplatins;

Squalamines;

Streptonigrins;

Streptovarycins;

Streptozocins;

Sufosfamides;

Sulofenurs;

Sunitinib Malate;

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 A'"'"'s (Guamecyclines);

Zeniplatins;

Zilascorbs [2-H];

Zinostatins;

Zoledronate;

Zorubicins; and

Zosuquidars.

25. A method for treating a hyaluronan-associated disease or condition, due to elevated interstitial fluid pressure, decreased vascular volume, elevated hyaluronan expression, and/or increased water content in a tissue, by administering to a subject the composition of claim 16, wherein the treating results in amelioration or reduction of one or more of increased interstitial fluid pressure, decreased vascular volume, and increased water content in a tissue.

26. A method for increasing penetration of therapeutic agents into tissues by administering to a subject the composition of claim 16.

27. The method of claim 25, wherein the disease or condition is benign prostatic hyperplasia.

28. The method of claim 27, wherein the esPH20 is PEGylated.

29. The esPH20 polypeptide of claim 1 that is N-partially glycosylated.

30. A combination, comprising:

a first composition containing an esPH20 polypeptide of claim 1; and

a second composition comprising a therapeutic agent.

31. The esPH20 polypeptide of claim 1 that consists of the sequence of amino acids set forth as amino acid residues 36-495, or a polypeptide that contains amino acid substitutions in the sequence of amino acids set forth as amino acid residues 36-495 of SEQ ID NO:

107, whereby the amino acid-substituted polypeptide consists of a sequence of amino acids that has at least 95% sequence identity with the sequence of amino acids set forth as amino acid residues 36-495 and exhibits hyaluronidase activity at neutral pH.

32. The esPH20 polypeptide of claim 1 that consists of the sequence of amino acids set forth as amino acid residues 36-496, or a polypeptide that contains amino acid substitutions in the sequence of amino acids set forth as amino acid residues 36-496 of SEQ ID NO:

107, whereby the amino acid-substituted polypeptide consists of a sequence of amino acids that has at least 95% sequence identity with the sequence of amino acids set forth as amino acid residues 36-496 and exhibits hyaluronidase activity at neutral pH.

33. The esPH20 polypeptide of claim 1 that consists of the sequence of amino acids set forth as amino acid residues 36-497, or a polypeptide that contains amino acid substitutions in the sequence of amino acids set forth as amino acid residues 36-497 of SEQ ID NO:

107, whereby the amino acid-substituted polypeptide consists of a sequence of amino acids that has at least 95% sequence identity with the sequence of amino acids set forth as amino acid residues 36-497 and exhibits hyaluronidase activity at neutral pH.

34. The esPH20 polypeptide of claim 1 that consists of the sequence of amino acids set forth as amino acid residues 36-498, or a polypeptide that contains amino acid substitutions in the sequence of amino acids set forth as amino acid residues 36-498 of SEQ ID NO:

107, whereby the amino acid-substituted polypeptide consists of a sequence of amino acids that has at least 95% sequence identity with the sequence of amino acids set forth as amino acid residues 36-498 and exhibits hyaluronidase activity at neutral pH.

35. The esPH20 polypeptide of claim 1 that consists of the sequence of amino acids set forth as amino acid residues 36-499, or a polypeptide that contains amino acid substitutions in the sequence of amino acids set forth as amino acid residues 36-499 of SEQ ID NO:

107, whereby the amino acid-substituted polypeptide consists of a sequence of amino acids that has at least 95% sequence identity with the sequence of amino acids set forth as amino acid residues 36-499 and exhibits hyaluronidase activity at neutral pH.

36. The esPH20 polypeptide of claim 1 that consists of the sequence of amino acids set forth as amino acid residues 36-500, or a polypeptide that contains amino acid substitutions in the sequence of amino acids set forth as amino acid residues 36-500 of SEQ ID NO:

107, whereby the amino acid-substituted polypeptide consists of a sequence of amino acids that has at least 95% sequence identity with the sequence of amino acids set forth as amino acid residues 36-500 and exhibits hyaluronidase activity at neutral pH.

37. The polypeptide of claim 1, wherein the mammalian cell is a CHO cell.