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Modified Recombinant Vaccinia Viruses, Uses Thereof

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Modified Recombinant Vaccinia Viruses, Uses Thereof (19) & (11) EP 2 028 270 B9 (12) CORRECTED EUROPEAN PATENT SPECIFICATION (15) Correction information: (51) Int Cl.: Corrected version no 1 (W1 B1) C12N 7/04 (2006.01) A61K 35/76 (2006.01) Corrections, see C12N 15/863 (2006.01) A61K 35/74 (2006.01) (2006.01) Description Paragraph(s) 238 C12Q 1/02 (48) Corrigendum issued on: 25.04.2012 Bulletin 2012/17 (45) Date of publication and mention of the grant of the patent: 15.02.2012 Bulletin 2012/07 (21) Application number: 08019998.7 (22) Date of filing: 18.06.2004 (54) Modified recombinant vaccinia viruses, uses thereof Modifizierte, rekombinante Vacciniaviren und deren Verwendungen Virus de vaccinia recombinés modifiés et utilisations associées (84) Designated Contracting States: (74) Representative: Blance, Stephen John AT BE BG CH CY CZ DE DK EE ES FI FR GB GR Boult Wade Tennant HU IE IT LI LU MC NL PL PT RO SE SI SK TR Verulam Gardens Designated Extension States: 70 Gray’s Inn Road AL HR LT LV MK London WC1X 8BT (GB) (30) Priority: 18.06.2003 EP 03013826 (56) References cited: 14.08.2003 EP 03018478 • GNANT M F ET AL: "Systemic administration of 22.10.2003 EP 03024283 a recombinant vaccinia virus expressing the cytosine deaminase gene and subsequent (43) Date of publication of application: treatment with 5-fluorocytosine leads to tumor- 25.02.2009 Bulletin 2009/09 specific gene expression and prolongation of survival in mice" CANCER RESEARCH, (62) Document number(s) of the earlier application(s) in AMERICAN ASSOCIATION FOR CANCER accordance with Art. 76 EPC: RESEARCH, BALTIMORE, MD.; US, vol. 59, no. 04816752.2 / 1 644 492 14, 15 July 1999 (1999-07-15), pages 3396-3403, XP009106317 (73) Proprietor: Genelux Corporation • MCCART J ANDREA ET AL: "Systemic cancer San Diego, California 92109 (US) therapy with a tumor-selective vaccinia virus mutant lacking thymidine kinase and vaccinia (72) Inventors: growth factor genes" CANCER RESEARCH, vol. • Szalay, Aladar 61, no. 24, 15 December 2001 (2001-12-15), pages Highland, CA 92346 (US) 8751-8757, XP002331814 ISSN: 0008-5472 • Timiryasova, Tatyana • TIMIRYASOVA TATYANA M ET AL: Scotrun, PA 18355 (US) "Construction of recombinant vaccinia viruses • Yu, Yong A. using PUV-inactivated virus as a helper" San Diego, CA 92130 (US) BIOTECHNIQUES, vol. 31, no. 3, September 2001 • Zhang, Qian (2001-09), pages 534-540, XP008022947 ISSN: San Diego, CA 92130 (US) 0736-6205 Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 028 270 B9 Printed by Jouve, 75001 PARIS (FR) (Cont. next page) EP 2 028 270 B9 • TIMIRYASOVA ET AL: "VISUALIZATION OF • J. NUCL. MEDICINE, vol. 34, no. 7, 1993, pages VACCINIA VIRUS INFECTION USING THE 1140-1146, RENILLA-LUCIFERASE-GFP FUSION PROTEIN" • J. BIOL. CHEM., vol. 274, no. 49, 1999, pages PROCEEDINGS OF THE 11TH INTERNATIONAL 34838-34845, SYMPOSIUM ON BIOLUMINESCENCE AND CHEMILUMINESCENCE, 2000, pages 457-460, XP008023166 2 EP 2 028 270 B9 Description FIELD OF THE INVENTION 5 [0001] Vaccines that contain attenuated or modified vaccinia virus , and methods for preparing vaccines are provides. In particular, vaccinia viruses are provided and pharmaceutical compositions thereof for treatment of proliferative and inflammatory disorders and for production of products in tumors are provided. BACKGROUND 10 [0002] In the late 19th century, a variety of attempts were made to treat cancer patients with microorganisms. One surgeon, William Coley, administered live Streptococcus pyogenes to patients with tumors with limited success. In the early 20th century, scientists documented vaccinia viral oncolysis in mice, which led to administration of several live viruses to patients with tumors from the 1940s through the 1960s. These forays into this avenue of cancer treatment 15 were not successful. [0003] Since that time, a variety of genetically engineered viruses have been tested for treatment of cancers. In one study, for example, nude mice bearing nonmetastatic colon adenocarinoma cells were systemically injected with a WR strain of vaccinia virus modified by having a vaccinia growth factor deletion and an enhanced green fluorescence protein inserted into the thymidine kinase locus. The virus was observed to have antitumor effect, including one complete 20 response, despite a lack of exogenous therapeutic genes in the modified virus (McCart et al. (2001) Cancer Res 61: 8751-8757). In another study, vaccinia melanoma oncolysate (VMO) was injected into sites near melanoma positive lymph nodes in a Phase III clinical trial of melanoma patients. As a control, New York City Board of Health strain vaccinia virus (VV) was administered to melanoma patients. The melanoma patients treated with VMO had a survival rate better than that for untreated patients, but similar to patients treated with the VV control (Kim et al. (2001) Surgical Oncol 10: 25 53-59). [0004] Other studies have demonstrated limited success with this approach. This therapy is not completely effective, particularly for systemically delivered viruses or bacteria. Limitations on the control of microbial vehicle function in vivo result in ineffective therapeutic results as well as raising safety concerns. It would be desirable to improve this type of therapy or to develop more effective approaches for treatments of neoplastic disease. Therefore, among the objects 30 herein, it is an object to provide microorganisms for the treatment of neoplastic and other diseases. SUMMARY [0005] Provided herein are pharmaceutical compositions and vaccinia viruses as described in the claims, for uses in 35 the treatment of neoplastic diseases and other diseases. Diseases for treatment are those in which the targeted tissues and/or cells are immunoprivileged in that they, and often the local environment thereof, somehow escape or are inac- cessible to the immune system. Such tissues include tumors and other tissues and cells involved in other proliferative disorders, wounds and other tissues involved in inflammatory responses. The viruses are selected or are designed to be non-pathogenic and to preferentially accumulate in the immunoprivileged tissues. The viruses, once in the tissues 40 or cells or vicinity thereof, affect the cell membranes of the cells in such tissues so that they become leaky or lyse, but sufficiently slowly so that the targeted cells and tumors leak enough antigen or other proteins for a time sufficient to elicit an immune response. [0006] The viruses are administered by any route, including systemic administration; such as i.v. or using oral or nasal or other delivery systems that direct agents to the lymphatics. In exemplary methods, the viruses are used to treat tumors 45 and to prevent recurrence and metastatic spread. The viruses are optionally modified to deliver other products, including other therapeutic products to the targeted tissues. [0007] When the vaccinia virus are administered to a host that contains tumors, the tumors in the host essentially become antigen and protein factories. This can be exploited so that the tumors can be used to produce proteins or other cellular products encoded by or produced by the virus. 50 [0008] Thus provided are vaccinia virus for elimination of immunoprivileged cells or tissues, particularly tumors. Virus according to the claims is administered typically systemically, and preferentially accumulates in immunoprivileged cells, such as tumor cells, resulting in leakage proteins and other compounds, such as tumor antigens, resulting in vaccination of the host against non-host proteins and, such as the tumor antigens, providing for elimination of the immunoprivileged cells, such as tumor cells, by the host’s immune system. The viruses are selected not for their ability to rapidly lyse cells, 55 but rather for the ability to accumulate in immunoprivileged cells, such as tumors, resulting in a leakage of antigens in a sufficient amount and for a sufficient time to elicit an immune response. [0009] Hence provided are vaccinia virus containing heterologous DNA, polypeptides or RNA to induce autoimmuni- zation of an organism against a tumor. In particular, the virus is selected or designed to accumulate in tumors and to 3 EP 2 028 270 B9 accumulate very little, if at all (to be non-toxic to the host) in non-tumorous cells, tissues or organs, and to in some manner result in the tumor cell lyses or cell membrane disruption such that tumor antigens leak. Such viruses are the LIVP-derived vaccinia virus described in the claims. The virus can be modified to express heterologous products that mediate or increase the leakage of the-tumor cell antigens and/or that are therapeutic, such as anti- tumor compounds. 5 [0010] Provided are attenuated vaccinia virus that accumulate in immunoprivileged tissues and cells, such as tumor cells, but do not accumulate to toxic levels in non-targeted organs and tissues, and that upon administration to an animal bearing the immunoprivileged tissues and cells, result in autoimmunity, such as by production of anti-tumor (or anti- tumor antigen) antibodies against the immunoprivileged cells or products thereof. The vaccinia viruses of the invention render the immunoprivileged cells leaky, such as by a slow lysis or apoptotic process. The goal is to achieve such 10 leakiness, but to not lyse the cells so rapidly that the host cannot mount an immune response. [0011] Vaccinia virus for eliminating immunoprivileged tissues and cells are provided. The viruses optionally include reporter genes and/or other heterologous nucleic acids that disrupt genes in the microorganism and can also encode and provide therapeutic products or products, such as RNA, including RNAi, that alter gene and/or protein expression in the cells or tissues where the microorganism accumulates.
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