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(12) United States Patent (10) Patent No.: US 8,039,240 B2 Roiz Et Al

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(12) United States Patent (10) Patent No.: US 8,039,240 B2 Roiz Et Al US008O39240B2 (12) United States Patent (10) Patent No.: US 8,039,240 B2 Roiz et al. (45) Date of Patent: Oct. 18, 2011 (54) RECOMBINANT HUMAN T2 RNASE AND 5,281.521 A 1/1994 Trojanowski et al. USES THEREOF 5,786.457 A 7, 1998 Nett et al. 6,590,075 B2 7/2003 Ruben et al. 7,101,839 B1 9, 2006 Roiz et al. (75) Inventors: Levava Roiz, Givat-Shmuel (IL); Betty 2005/O113327 A1 5/2005 Roiz et al. Schwartz, Rechovot (IL); Patricia 2008, OO13419 A1 1/2008 Lee et al. Smirnoff, Rehovot (IL); Oded Shoseyov, Karme Yossef (IL) FOREIGN PATENT DOCUMENTS AU T69011 3, 2001 EP 1207 755 5, 2002 (73) Assignee: Yissum Reseach Development WO WO99,33981 7, 1999 Company of the Hebrew University of WO WOO1/15531 8, 2001 Jerusalem, Jerusalem (IL) OTHER PUBLICATIONS (*) Notice: Subject to any disclaimer, the term of this Smirnoff et al., Cancer 107(12):2760-2769; 2006.* patent is extended or adjusted under 35 Garret et al., European Journal of Cancer 35(14): 2010-2030, 1999.* U.S.C. 154(b) by 834 days. Deshpande et al., Critical Reviews in Microbiology 28(2):79-122. 20O2.* (21) Appl. No.: 11/664,132 Branden et al., Introduction to Protein Structure, Garland Publishing Inc., New York, p. 247, 1991.* PCT Fled: Sep. 28, 2005 Seffernicket al., J. Bacteriol. 183(8):2405-2410, 2001.* (22) Witkowski et al. Biochemistry 38: 11643-1 1650, 1999.* PCT NO.: PCT/L2OOS/OO1041 Guo et al., PNAS 101(25):9205-9210, 2004.* (86) Trubia et al., GenBank accession No. NP 003721, Jun. 28, 1999.* S371 (c)(1), Roiz et al. “Stigmatic RNase in Self-Compatible Peach (Prunus (2), (4) Date: Nov. 6, 2007 persica)”. International Journal of Plant Sciences, 156:(1): 37-41, 1995. PCT Pub. No.: WO2O06/035439 Zhu et al. “Escherichia coli RNA Gene Encoding RNase I: Cloning, (87) Overexpression, Subcellular Distribution of the Enzyme, and Use of PCT Pub. Date: Apr. 6, 2006 an RNA Deletion to Identify Additional RNases”. Journal of Bacte riology, 172(6): 3146-3151, 1990. (65) Prior Publication Data Ye et al. “Isolation and Characterization of cDNAs Encoding Zylogenesis-Associated and Wounding-Induced Ribonucleases in US 2008/O131419 A1 Jun. 5, 2008 Zinnia elegans”. Plant Molecular Biology, 30: 697-709, 1996. Irie et al. “Role of Histidine 46 in the Hydrolysis and the Reverse Related U.S. Application Data Transphosphorylation Reaction of RNase Rh From Rhizopus niveus”, Journal of Biochemistry, 121: 849-853, 1997. (60) Provisional application No. 60/613,702, filed on Sep. Laemmli"Relevant Page on Gel Electrophoresis', Nature, 227: 681, 29, 2004. 1970. Abstract. Broothaerts et al. “Petunia Hybrida S-Proteins: Ribonuclease Activ (51) Int. C. ity and the Role of Their Glycan Side Chains in Self-Incompatibil A6 IK38/46 (2006.01) ity”, Sexual Plant Reproduction, 4:258-266, 1991. CI2O I/44 (2006.01) Bradford "A Rapid and Sensitive Method for the Quantitation of CI2N 9/16 (2006.01) Microgram Quantities of Protein Utilizing the Principle of Protein CI 2N 9/22 (2006.01) Dye Binding”. Analytical Biochemistry, 72: 248-254, 1976. CO7K (4/OO (2006.01) Yokota et al. “Isolation and Characterization of Plant Myosin From Pollen Tubes of Lily”. Protoplasma, 177: 153-162, 1994. (52) U.S. C. ......... 435/196; 424/94.6:435/199: 435/19; Lin et al. “Characterization of the Mechanism of Cellular and Cell 530/350 Free Protein Synthesis Inhibition by an Anti-Tumor Ribonuclease'. (58) Field of Classification Search .................. 435/196, Biochemical and Biophysical Research Communications, 204(1): 435/199, 19; 424/94.6; 530/350 156-162, 1994. See application file for complete search history. (Continued) (56) References Cited Primary Examiner — Delia Ramirez U.S. PATENT DOCUMENTS (57) ABSTRACT 3,791932 A 2f1974 Schuurs et al. 3,839,153 A 10, 1974 Schuurs et al. The cloning and heterologous expression of human T2 3,850,578 A 11/1974 McConnell RNase6PL are disclosed. Further disclosed are methods for 3,850,752 A 11/1974 Schuurs et al. preventing, inhibiting and/or reversing cell motility, actin 3,853.987 A 12/1974 Dreyer 3,867,517 A 2/1975 Ling filament assembly or disassembly, proliferation, coloniza 3,879,262 A 4, 1975 Schuurs et al. tion, differentiation, accumulation and/or development of 3,901,654 A 8, 1975 Gross abnormal cells in a subject is disclosed. The methods are 3,935,074 A 1/1976 Rubenstein et al. effected by administering to the subject a therapeutically 3,984,533 A 10/1976 Uzgiris effective amount of a RNase6PL ribonuclease or close homo 3.996,345 A 12/1976 Ullman et al. 4,034,074 A T. 1977 Miles logues thereof. 4,098,876 A 7, 1978 Piasio et al. 4,879,219 A 11, 1989 Wands et al. 5 Claims, 49 Drawing Sheets 5,011,771 A 4, 1991 Bellet et al. (22 of 49 Drawing Sheet(s) Filed in Color) US 8,039,240 B2 Page 2 OTHER PUBLICATIONS Roiz et al. “Stigmatic Rnase in Calamondin (Citrus reticulata Var. Brown et al. “Barley Aleurone Layers Secrete a Nuclease in Austera X Fortunella Sp.)”, Physiologia Plantarum, 94: 585-590, Response to Gibberellic Acid', Plant Physiology, 82: 801-806, 1986. 1995. Heslop-Harrison et al. "Dynamic Aspects of Apical Zonation in the Laccetti et al. “Seminal Ribonuclease Inhibits Tumor Growth and Angiosperm Pollen Tube'. Sexual Plant Reproduction, 3: 187-194, Reduces the Metastatic Potential of Lewis Lung Carcinoma'. Cancer 1990. Research, 54:4523-4526, 1994. Irie et al. “Site of Alkylation of the Major Ribonuclease From Trubia et al. “Mammalian Rh/T2/S-Glucoprotein Ribonuclease Fam Aspergillus saitoi With Lodoacetate'. Journal of Biochemistry, ily Genes: Cloning of a Human Member in a Region of Chromosome 99(3): 627-633, 1986. 6 (6q27) Frequently Deleted in Human Malignancies'. Genomics, Simm et al. “On the Interaction of Bovine Seminal Rinase With Actin 42(2): 342-344, 1997. Abstract. in Vitro', European Journal of Biochemistry, 166: 49-54, 1987. Deshpande etal. “Ribonucleases From T2 Family”. Critical Reviews Kawata et al. “Amino-Acid Sequence of Ribonuclease T2 From in Microbiology, 28(2): 79-122, 2002. Aspergillus oryzae'. European Journal of Biochemistry, 176: 683 Castro et al. “Gene Therapy for Parkinson's Disease: Recent 697, 1988, p. 688, Claims 15, 20. Achievements and Remaining Challenges'. Histology and Ohgi et al. “Expression of Rnase Rh From Rhizopus niveus in Yeast Histopathology, 16: 1225-1238, 2001. and Characterization of the Secreted Proteins”, Journal of Biochem Acquati et al. “Tumor and Metastasis Suppression by the Human istry, 109: 776-785, 1991, p. 777, Claims 21, 26. RNASET2 Gene'. International Journal of Oncology, 26(5): 1159 Irie et al. “Structure-Function Relationships of Acid Ribonucleases: 1168, 2005. Abstract. Lysosomal, Vacuolar and Periplasmic Enzymes'. Pharmacological Mastronicola et al. “Key Extracellular and Intracellular Steps in the Therapy, 81(2): 77-89, 1999. p. 79, Claims 15, 20. Antitumor Action of Seminal Ribonuclease”, European Journal of Roiz et al. “Characterization of Aspergillus niger B-1 RNase and Its Biochemistry, 230(1): 242-249, 1995. Inhibitory Effect on Pollen Germination and Pollen Tube Growth in Leland et al. “Cancer Chemotherapy—Ribonucleases to the Res Selected Tree Fruit”, Journal of the American Society of Horticulture cue”. Chemistry & Biology, 8: 405-413, 2001. Sciences, 125(1): 9-14, 2000. Abstract. Ide et al. “The Complete Amino Acid Sequence of Ribonuclease Nomachi et al. “Purification and Some Properties of 2 Acid RNase From Seeds of Bitter Gourd (Momortia charantia)”. FEBS Letters, Fractions EC-3.1.27.1 EC-3.1.27.3 From the Mycelia of Aspergillus 284(2): 161-164, 1991. niger”, Journal of General and Applied Microbiology, 26(6): 375 Appellant's Brief on Appeal Under 37 CFRS 41.37 Dated Mar. 22. 386, 1980. 2010 in the US Patent and Trademark Office Re.: U.S. Appl. No. Hu et al. “Technology to Obtain Sustained Release Characteristics of 10/952,495. Drugs. After Delivered to the Colon”, Journal of Drug Targeting, 6(6): Communication Pursuant to Article 94(3) EPC Dated Feb. 12, 2010 439-448, 1999. Abstract. From the European Patent Office Re.: Application No. 05789520.3. Ishibashi et al. “Evaluation of Colonic Absorbability of Drugs in Notice of Appeal in Reponse Dated Jan. 26, 2010 to Official Action of Dogs. Using a Novel Colon-Targeted Delivery Capsule (CTDC)”, Aug. 5, 2009 From the US Patent and Trademark Office Re. U.S. Journal of Control Release, 59(3): 361-376, 1999. Abstract. Appl. No. 10/952,495. Matsuda et al. “Effect of Food Intake on the Delivery of Fluorescein Office Action Dated Dec. 7, 2009 From the Israel Patent Office Re.: as a Model Drug in Colon Delivery Capsule After Oral Administra Applicaiton No. 199252 and Its Translation Into English. tion to Beagle Dogs”, Journal of Drug Targeting, 4(2): 59-67, 1996. Office Action Dated Feb. 16, 2010 From the Israel Patent Office Re.: Abstract. Application No. 182323 and Its Translation Into English. Takaya et al. “Development of a Colon Delivery Capsule and the Office Action Dated Feb. 16, 2010 From the Israel Patent Office Re.: Pharmacological Activity of Recombinant Human Granulocyte Application No. 199252 and Its Translation Into English. Colony-Stimulating Factor (RhC-CSF) in Beagle Dogs”, Journal of Official Action Dated Nov. 5, 2003 From the US Patent and Trade Pharmacy and Pharmacology, 47(6):474-478, 1995. Abstract. mark Office Re.: U.S. Appl. No. 10/069,454. Tozaki et al. "Chitosan Capsules for Colon-Specific Drug Delivery: Official Action Dated Jan. 12, 2005 From the US Patent and Trade Improvement of Insulin Absorption From the Rat Colon”, Journal of mark Office Re.: U.S.
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