(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`(19) World Intellectual Property
`Organization
`International Bureau
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`(43) International Publication Date
`21 January 2016 (21.01.2016)
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`WIPOI PCT
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`\9
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`(10) International Publication Number
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`WO 2016/008976 A1
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`(51)
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`International Patent Classification:
`A61K 35/768 (2015.01)
`C07K 16/18 (2006.01)
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`(21)
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`International Application Number:
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`PCT/EP2015/066263
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`(22)
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`International Filing Date:
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`(25)
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`(26)
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`(30)
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`(71)
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`(72)
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`(74)
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`(81)
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`Filing Language:
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`Publication Language:
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`16 July 2015 (16.07.2015)
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`English
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`English
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`Priority Data:
`143061539
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`16 July 2014 (16.07.2014)
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`EP
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`Applicant: TRANSGENE SA [FR/FR]; Parc d'Innovation
`Boulevard Gonthier d'Andernach, F-67400 lllkirch Graf-
`fenstaden (FR).
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`Inventors: SILVESTRE, Nathalie; 6, rue des Iris, F-
`67120 Ergersheim (FR). GEIST, Michel; 18, rue Edouard
`Manet, F—67l70 Brumath (FR). RITTNER, Karola; 51,
`rue des Pluviers, F-67100 Strasbourg (FR). MARCHAND,
`Jean—Baptiste;
`1A rue des Hauts Paturages, F—67210
`Obernai (FR). THIOUDELLET, Christine; 4, boulevard
`dc la Victoirc, F-67000 Strasbourg (FR).
`
`Agent: REGIMBEAU; 20, rue de Chazelles, F-75847 Par-
`is Cedex 17 (FR).
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`Designated States (unless otherwise indicated, for every
`kind ofnational protection available): AE, AG, AL, AM,
`
`A0, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY,
`BZ, CA, CII, CL, CN, CO, CR, CU, CZ, DE, DK, DM,
`DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT,
`IIN, IIR, IIU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR,
`KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG,
`MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM,
`PA, PE, PG, PH, PL, PT, QA, R0, RS, RU, RW, SA, SC,
`SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN,
`TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW.
`
`(84)
`
`Designated States (unless otherwise indicated, for every
`kind of regional protection available): ARIPO (BW, GH,
`GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ,
`TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU,
`TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE,
`DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU,
`LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK,
`SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ,
`GW, KM, ML, MR, NE, SN, TD, TG).
`Declarations under Rule 4.17:
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`ofinventorship (Rule 4.17(iv))
`Published:
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`with international search report (Art. 21(3))
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`with sequence listing part ofa’escription (Rule 5.2(a))
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`(54) Title: ONCOLYTIC VIRUS FOR EXPRESSION OF IMMUNE CHECKPOINT MODULATORS
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`(57) Abstract: The present invention provides an oncolytic virus comprising nucleotide sequence(s) encoding one or more immune
`checkpoint modulator(s). It also concerns a pharmaceutical composition comprising effective amount of said oncolytic virus and,
`eventually, a pharmaceutically acceptable vehicle and its use for treating proliferative diseases such as cancers.
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`Oncolytic virus for expression of immune checkpoint modulators
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`Field of the invention
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`The present invention generally relates to the field of oncolytic virotherapy and more
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`specifically to compositions and methods to treat, prevent, or
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`inhibit proliferative diseases,
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`especially cancer. Embodiments include an oncolytic virus comprising nucleotide sequence(s)
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`encoding one or more
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`immune
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`checkpoint modulator(s). Embodiments
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`also include
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`a
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`pharmaceutical composition comprising such oncolytic virus and, eventually, a pharmaceutically
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`acceptable vehicle and its use for treating proliferative diseases such as cancers.
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`Cancer is caused by both external factors (e.g. tobacco, infectious organisms, alimentary
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`habits, chemicals, and radiation) and internal factors (e.g. inherited mutations, hormones, immune
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`conditions, and mutations that occur from metabolism). Each year, cancer is diagnosed in more
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`than 12 million subjects worldwide.
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`In industrialized countries, approximately one person out five
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`will die of cancer. Although a vast number of chemotherapeutics exist, they are often ineffective,
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`especially against malignant and metastatic tumors that establish at a very early stage of the
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`disease. Moreover, antitumor immunity is often ineffective clue to the fact that tumor cells have
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`evolved mechanisms to escape host defense. One of the major mechanisms of immune suppression
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`is a process known as "T—cell exhaustion”, which results from chronic exposure to antigens and is
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`characterized by the upregulation of inhibitory receptors. These inhibitory receptors serve as
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`immune checkpoints in order to prevent uncontrolled immune reactions. Various
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`immune
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`checkpoints acting at different levels of T cell immunity have been described in the literature,
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`including programmed cell death protein 1 (PD—1) and its ligands PD—Ll and PD—L2, CTLA—4 (cytotoxic
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`T—lymphocyte associated protein—4), LAGB, B and T lymphocyte attenuator, T—cell immunoglobulin,
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`mucin domain—containing protein 3 (TIM—3), and V—domain immunoglobulin suppressor of T cell
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`activation.
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`Whatever the mechanism of action, these immune checkpoints can inhibit the development
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`of an efficient anti-tumor immune response. There is increasing interest in the possible therapeutic
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`benefits of blocking such immune checkpoints as a means of inhibiting immune system tolerance to
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`tumors and thus rescue exhausted antitumor T cells (Leach et al., 1996, Science 271: 1734-6). A vast
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`number of antagonistic antibodies have been developed during the last decade (e.g. anti Tim3, -PD-
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`L1,
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`-CTL/—\-4, -PD1, etc) and most importantly, some have been associated with objective clinical
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`responses in cancer patients. Antibodies targeting CTLA-4 are already marketed (e.g. Ipilimumab,
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`Yervoy, Bristol-Myers Squibb) for metastatic melanoma. BMS reported that from 1800 melanoma
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`patients treated with ipilimumab 22% are still alive 3 years later. Antibody therapies with anti PD-L1
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`(e.g. MPDL3280A, Roche), anti PD-1 (e.g. Nivolumab, BMS) are also ongoing.
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`Another therapeutic approach that is emerging in the field of cancer is oncolytic viruses
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`(Hermiston, 2006, Curr. Opin. Mol. Ther. 8: 322-30). Oncolytic viruses are capable of selective
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`replication in dividing cells (e.g. cancer cell) while leaving non dividing cells (e.g. normal cells)
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`unharmed. As the infected dividing cells are destroyed by lysis, they release new infectious virus
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`particles to infect the surrounding dividing cells. Cancer cells are ideal hosts for many viruses
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`because they have the antiviral interferon pathway inactivated or have mutated tumour suppressor
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`genes that enable viral replication to proceed unhindered (Chernajovsky et al., 2006, British Med. J.
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`332: 170-2). A number of viruses including adenovirus,
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`reovirus, measles, herpes simplex,
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`Newcastle disease virus and vaccinia have now been clinically tested as oncolytic agents.
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`Some viruses are naturally oncolytic (such as reovirus and the Seneca valley picornavirus)
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`while others are engineered for tumor selectivity by modifying the viral genome. Such modifications
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`include functional deletions in essential viral genes, the use of tumor- or tissue-specific promoters
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`to control the viral gene expression and tropism modification to redirect virus to the cancer cell
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`surface.
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`The first oncolytic virus to be approved by a regulatory agency was a genetically modified
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`adenovirus named H101 (Shanghai Sunway Biotech) that gained approval in 2005 from China's State
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`Food and Drug Administration (SFDA) for the treatment of head and neck cancer. Another oncolytic
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`adenovirus, named ONYX—015 is in ongoing clinical trials for the treatment of various solid tumors
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`(in phase III for the treatment of recurrent head and neck cancer) (Cohen et al., 2001, Curr. Opin.
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`Investig. Drugs 2: 1770—5). As another example, oncolytic herpes simplex 1 (T—VEC) was genetically
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`engineered to attenuate the virus virulence,
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`increase selectivity for cancer cells and enhance
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`antitumor
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`immune response (through GM—CSF expression). Clinical efficacy in unresectable
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`melanoma has been demonstrated in Phase II and Phase III clinical trials (Senzer et al, 2009, J. Clin.
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`Oncol. 27: 5763-71).
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`Vaccinia viruses (VV) possess many of the key attributes necessary for use in oncolytic
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`virotherapy such as natural tropism for tumors, strong Iytic ability, short life cycle with rapid cell-to-
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`cell spread, highly efficient gene expression and a large cloning capacity. In addition, they have been
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`delivered to millions of individuals during the smallpox eradication campaign without major safety
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`concerns.
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`In this respect, a TK and VGF double deleted VV (Wyeth strain) expressing GM-CSF
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`(named JX-963) showed significant cancer selectivity in tumor bearing mice (Thorne et al., 2007, J
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`Clin Invest. 117: 3350-8). On the same line, JX-594, a TK-deleted VV (Wyeth strain) armed with GM-
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`CSF, has shown promising clinical data, and a randomized Phase III trial in hepatocellular carcinoma
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`is expected to start soon.
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`Combination therapies involving oncolytic virus and immune checkpoint inhibitors have
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`been described in the literature. W02014/022138 describes the combination of irradiated tumor
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`cells, an oncolytic adenovirus and an anti CTLA4 antibody for use for treating bladder or prostate
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`cancer. W02014/O47350 envisages a recombinant oncolytic virus with a gene encoding an anti-PD-
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`1 antibody inserted in the viral genome without providing any working example that would support
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`utility of such an oncolytic virus.
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`Technical problem
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`One may expect that cancer will continue to be a serious global health threat for many
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`years due to the high number of causal factors that may act together or separately to initiate or
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`promote the development of a cancer. Moreover, malignant and especially metastatic tumors are
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`often resistant to conventional therapies explaining the significant morbidity of some cancers.
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`Thus, there is an important need to develop more effective approaches, for improving
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`prevention and treatment of such proliferative diseases, and especially metastatic cancers. The
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`present invention provides a unique product combining oncolysis for killing dividing cells and
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`immune checkpoint for breaking cancer-associated immune tolerance.
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`This technical problem is solved by the provision of the embodiments as defined in the
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`claims.
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`Other and further aspects, features and advantages of the present
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`invention will be
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`apparent from the following description of the presently preferred embodiments of the invention.
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`These embodiments are given for the purpose of disclosure.
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`Summary of the invention
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`The present invention concerns an oncolytic virus comprising inserted in its genome one or
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`more nucleic acid molecule(s) encoding one or more immune checkpoint modulator(s).
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`The oncolytic virus is preferably selected from the group consisting of reovirus, New Castle
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`Disease virus (NDV), vesicular stomatitis virus (VSV), measles virus,
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`influenza virus, Sinbis virus,
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`adenovirus, poxvirus and herpes virus (HSV) and the like. In one embodiment, the oncolytic virus is
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`a vaccinia virus.
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`In a preferred embodiment, the vaccinia virus is engineered to lack thymidine
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`kinase activity (e.g. the genome of said W has an inactivating mutation in JZR gene to produce a
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`defective TK phenotype). Alternatively or in combination, the vaccinia virus is engineered to lack RR
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`activity (e.g. the genome of said VV has an inactivating mutation in |4L and/or F4L gene to produce
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`a defective RR phenotype).
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`In one embodiment, the vaccinia virus further expresses at least one therapeutic gene, in
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`particular a gene encoding a suicide gene product and/or an immunostimulatory protein.
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`In one embodiment, the encoded one or more immune checkpoint modulator(s) is an
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`antagonist molecule that antagonizes the activity of PD-l, PD-Ll or CTLA4 with a specific preference
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`for an anti PD-l antibody and/or an anti CTLA4 antibody.
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`The present
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`invention further provides a composition comprising said oncolytic virus,
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`eventually with a pharmaceutical acceptable vehicle.
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`In one embodiment,
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`the composition is
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`formulated for intravenous or intratumoral administration.
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`The present invention also concerns the use of said oncolytic virus or composition thereof
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`for treating a proliferative disease as well as a method of treatment relying on the administration of
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`an effective amount of said oncolytic virus or composition thereof.
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`In one embodiment,
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`the
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`proliferative disease treated by the method of the invention is cancer and especially melanoma,
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`renal cancer, prostate cancer, breast cancer, colorectal cancer, lung cancer and liver cancer. In one
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`embodiment,
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`the use or method comprises an additional step in which a pharmaceutically
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`acceptable amount of a prodrug is administered to said mammal. The administration of said
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`prodrug takes place preferably at least 3 days after the administration of said oncolytic virus or virus
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`composition.
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`Detailed description
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`The present invention concerns an oncolytic virus comprising inserted in its genome one or
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`more nucleic acid molecule(s) encoding one or more immune checkpoint modulator(s).
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`Definitions
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`As used throughout the entire application, the terms "a" and "an" are used in the sense that
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`they mean "at least one", "at least a first", "one or more" or "a plurality" of the referenced
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`components or steps, unless the context clearly dictates otherwise. For example, the term "a cell"
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`includes a plurality of cells, including mixtures thereof.
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`The term ”one or more” refers to either one or a number above one (e.g. 2, 3, 4, 5, etc).
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`The term "and/or" wherever used herein includes the meaning of "and", "or" and "all or any
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`other combination of the elements connected by said term".
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`The term "about" or "approximately" as used herein means within 20%, preferably within
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`10%, and more preferably within 5% of a given value or range.
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`As used herein, when used to define products, compositions and methods,
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`the term
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`"comprising" (and any form of comprising, such as "comprise" and "comprises"), "having" (and any
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`form of having, such as "have" and "has"), "including" (and any form of including, such as "includes"
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`and "include") or "containing" (and any form of containing, such as "contains" and "contain") are
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`open-ended and do not exclude additional, unrecited elements or method steps. Thus,
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`a
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`polypeptide "comprises" an amino acid sequence when the amino acid sequence might be part of
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`the final amino acid sequence of the polypeptide. Such a polypeptide can have up to several
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`hundred additional amino acids residues. "Consisting essentially of" means excluding other
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`components or steps of any essential significance. Thus, a composition consisting essentially of the
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`recited components would not exclude trace contaminants and pharmaceutically acceptable
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`carriers. A polypeptide "consists essentially of” an amino acid sequence when such an amino acid
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`sequence is present with eventually only a few additional amino acid residues. "Consisting of”
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`means excluding more than trace elements of other components or steps. For example, a
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`polypeptide "consists of” an amino acid sequence when the polypeptide does not contain any
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`amino acids but the recited amino acid sequence.
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`The terms ”polypeptide”, ”peptide” and ”protein” refer to polymers of amino acid residues
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`which comprise at least nine or more amino acids bonded via peptide bonds. The polymer can be
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`linear, branched or cyclic and may comprise naturally occurring and/or amino acid analogs and it
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`may be interrupted by non—amino acids. As a general indication, if the amino acid polymer is more
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`than 50 amino acid residues, it is preferably referred to as a polypeptide or a protein whereas if it is
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`50 amino acids long or less, it is referred to as a "peptide”.
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`Within the context of the present
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`invention,
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`the terms ”nucleic acid”, ”nucleic acid
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`molecule”, ”polynucleotide” and "nucleotide sequence” are used interchangeably and define a
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`polymer of any length of either polydeoxyribonucleotides (DNA)
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`(e.g. cDNA, genomic DNA,
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`plasmids, vectors, viral genomes,
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`isolated DNA, probes, primers and any mixture thereof) or
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`polyribonucleotides
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`(RNA)
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`(e.g. mRNA,
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`antisense
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`RNA,
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`SiRNA)
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`or mixed
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`polyribo-
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`polydeoxyribonucleotides. They encompass single or double-stranded, linear or circular, natural or
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`synthetic, modified or unmodified polynucleotides. Moreover, a polynucleotide may comprise non-
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`naturally occurring nucleotides and may be interrupted by non-nucleotide components.
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`The term "analog" or ”variant” as used herein refers to a molecule (polypeptide or nucleic
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`acid) exhibiting one or more modification(s) with respect
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`to the native counterpart. Any
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`modification(s) can be envisaged, including substitution, insertion and/or deletion of one or more
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`nucleotide/amino acid residue(s). Preferred are analogs that retain a degree of sequence identity of
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`at least 80%, preferably at least 85%, more preferably at least 90%, and even more preferably at
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`least 98% identity with the sequence of the native counterpart.
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`In a general manner, the term ”identity" refers to an amino acid to amino acid or nucleotide
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`to nucleotide correspondence between two polypeptide or nucleic acid sequences. The percentage
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`of identity between two sequences is a function of the number of identical positions shared by the
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`sequences, taking into account the number of gaps which need to be introduced for optimal
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`alignment and the length of each gap. Various computer programs and mathematical algorithms
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`are available in the art to determine the percentage of identity between amino acid sequences,
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`such as for example the Blast program available at NCBI or ALIGN in Atlas of Protein Sequence and
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`Structure (Dayhoffed, 1981, Suppl., 3: 482-9). Programs for determining identity between
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`nucleotide sequences are also available in specialized data base (e.g. Genbank, the Wisconsin
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`Sequence Analysis Package, BESTFIT, FASTA and GAP programs). For illustrative purposes, ”at least
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`80% identity” means 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,
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`94%, 95%, 96%, 97%, 98%, 99% or 100%.
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`As used herein,
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`the term "isolated”
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`refers
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`to a protein, polypeptide, peptide,
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`polynucleotide, vector, etc., that is removed from its natural environment (i.e. separated from at
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`least one other component(s) with which it is naturally associated or found in nature). For example,
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`a nucleotide sequence is isolated when it is separated of sequences normally associated with it in
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`nature (e.g. dissociated from a genome) but it can be associated with heterologous sequences.
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`The term "obtained from", ”originating” or "originate” is used to identify the original source
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`of a component (e.g. polypeptide, nucleic acid molecule) but is not meant to limit the method by
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`which the component is made which can be, for example, by chemical synthesis or recombinant
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`means.
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`As used herein, the term ”host cell” should be understood broadly without any limitation
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`concerning particular organization in tissue, organ, or isolated cells. Such cells may be of a unique
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`type of cells or a group of different types of cells such as cultured cell
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`lines, primary cells and
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`dividing cells. In the context of the invention, the term ”host cells” include prokaryotic cells, lower
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`eukaryotic cells such as yeast, and other eukaryotic cells such as insect cells, plant and mammalian
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`(e.g. human or non-human) cells as well as cells capable of producing the oncolytic virus and/or the
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`immune checkpoint modulator(s) for use in the invention. This term also includes cells which can be
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`or has been the recipient of the vectors described herein as well as progeny of such cells.
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`As used herein, the term ”oncolytic virus” refers to a virus capable of selectively replicating
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`in dividing cells (e.g. a proliferative cell such as a cancer cell) with the aim of slowing the growth
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`and/or lysing said dividing cell, either in vitro or in vivo, while showing no or minimal replication in
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`non-dividing cells. Typically, an oncolytic virus contains a viral genome packaged into a viral particle
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`(or virion) and is infectious (i.e. capable of infecting and entering into a host cell or subject). As used
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`herein, this term encompasses DNA or RNA vector (depending on the virus in question) as well as
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`viral particles generated thereof.
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`The term ”treatment” (and any form of treatment such as ”treating”, "treat”) as used
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`herein encompasses prophylaxis (e.g. preventive measure in a subject at
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`risk of having the
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`pathological condition to be treated) and/or therapy (e.g.
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`in a subject diagnosed as having the
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`pathological condition), eventually in association with conventional therapeutic modalities. The
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`result of the treatment is to slow down, cure, ameliorate or control the progression of the targeted
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`pathological condition. For example,
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`a subject
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`is successfully treated for a cancer
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`if after
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`administration of an oncolytic virus as described herein,
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`the subject shows an observable
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`improvement of its clinical status.
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`The term ”administering” (or any form of administration such as ”administered”) as used
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`herein refers to the delivery to a subject of a therapeutic agent such as the oncolytic virus described
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`herein.
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`As used herein, the term "proliferative disease” encompasses any disease or condition
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`resulting from uncontrolled cell growth and spread including cancers as well as diseases associated
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`to an increased osteoclast activity (e.g. rheumatoid arthritis, osteoporosis, etc) and cardiovascular
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`diseases (restenosis that results from the proliferation of the smooth muscle cells of the blood
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`vessel wall, etc). The term ”cancer” may be used interchangeably with any of the terms ”tumor”,
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`”malignancy”, ”neoplasm”, etc. These terms are meant to include any type of tissue, organ or cell,
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`any stage of malignancy (e.g. from a prelesion to stage IV)
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`The term ”subject” generally refers to an organism for whom any product and method of
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`the invention is needed or may be beneficial. Typically, the organism is a mammal, particularly a
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`mammal selected from the group consisting of domestic animals, farm animals, sport animals, and
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`primates. Preferably, the subject is a human who has been diagnosed as having or at risk of having a
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`proliferative disease such as a cancer. The terms ”subject” and ”patients” may be used
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`interchangeably when referring to a human organism and encompasses male and female. The
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`subject to be treated may be a newborn, an infant, a young adult or an adult.
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`The term ”combination” or ”association” as used herein refers to any arrangement possible
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`of various components (e.g. an oncolytic virus and one or more substance effective in anticancer
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`therapy). Such an arrangement
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`includes mixture of said components as well as separate
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`combinations for concomitant or sequential administrations. The present invention encompasses
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`combinations comprising equal molar concentrations of each component as well as combinations
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`with very different concentrations. It is appreciated that optimal concentration of each component
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`of the combination can be determined by the artisan skilled in the art.
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`The term ”immune checkpoint modulator” refers to a molecule capable of modulating the
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`function of an immune checkpoint protein in a positive or negative way (in particular the interaction
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`between an antigen presenting cell (APC) or a cancer cell and a T effector cell). The term "immune
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`checkpoint” refers to a protein directly or indirectly involved in an immune pathway that under
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`normal physiological conditions is crucial for preventing uncontrolled immune reactions and thus
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`for
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`the maintenance of self-tolerance and/or tissue protection. The one or more immune
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`checkpoint modulator(s) in use herein may independently act at any step of the T cell-mediated
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`immunity including clonal selection of antigen-specific cells, T cell activation, proliferation,
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`trafficking to sites of antigen and inflammation, execution of direct effector function and signaling
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`through cytokines and membrane ligands. Each of these steps is regulated by counterbalancing
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`stimulatory and inhibitory signals that in fine tune the response.
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`In the context of the present
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`invention, the term encompasses immune checkpoint modulator(s) capable of down-regulating at
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`least partially the function of an inhibitory immune checkpoint (antagonist) and/or immune
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`checkpoint modulator(s) capable of up-regulating at least partially the function of a stimulatory
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`immune checkpoint (agonist).
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`20
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`Oncolytic virus
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`The oncolytic virus of the present invention can be obtained from any member of virus
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`identified at present time provided that it is oncolytic by its propensity to selectivity replicate and
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`kill dividing cells as compared to non—dividing cells.
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`It may be a native virus that is naturally
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`oncolytic or may be engineered by modifying one or more viral genes so as to increase tumor
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`selectivity and/or preferential replication in dividing cells, such as those involved in DNA replication,
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`nucleic acid metabolism, host tropism, surface attachment, virulence,
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`lysis and spread (see for
`
`example Kim et al., 2001, Nat. Med. 7: 781; Wong et al., 2010, Viruses 2: 78-106). One may also
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`envisage placing one or more viral gene(s) under the control of event or tissue-specific regulatory
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`elements (e.g. promoter).
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`Exemplary oncolytic viruses include without limitation reovirus, Seneca Valley virus (SVV),
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`vesicular stomatitis virus (VSV), Newcastle disease virus (NDV), herpes simplex virus (HSV),
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`morbillivirus virus, retrovirus, influenza virus, Sinbis virus, poxvirus, adenovirus, or the like.
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`In one embodiment, the oncolytic virus of the present invention is obtained from a reovirus.
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`A representative example
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`includes Reolysin
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`(under development
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`by Oncolytics Biotech;
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`NCT01166542).
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`In one embodiment, the oncolytic virus of the present invention is obtained from a Seneca
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`Valley virus. A representative example includes NTX-010 (Rudin et al., 2011, Clin. Cancer. Res. 17(4):
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`888-95).
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`In one embodiment, the oncolytic virus of the present invention is obtained from a vesicular
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`stomatitis virus (VSV). Representative examples are described in the literature (e.g. Stojdl et al.,
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`2000, Nat. Med. 6(7): 821-5; Stojdl et al., 2003, Cancer Cell 4(4): 263-75).
`
`In one embodiment,
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`the oncolytic virus of the present
`
`invention is obtained from a
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`Newcastle disease virus. Representative examples include without limitation the 73-T PV701 and
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`HDV-HUJ strains as well as those described in the literature (e.g. Phuangsab et al., 2001, Cancer
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`Lett. 172(1): 27-36; Lorence et al., 2007, Curr. Cancer Drug Targets 7(2): 157-67; Freeman et al.,
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`2006, Mol. Ther. 13(1): 221-8).
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`In one embodiment, the oncolytic virus of the present invention is obtained from a herpes
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`virus. The Herpesviridae are a large family of DNA viruses that all share a common structure and are
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`composed of relatively large double-stranded,
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`linear DNA genomes encoding 100-200 genes
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`encapsided within an icosahedral capsid which is enveloped in a lipid bilayer membrane. Although
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`the oncolytic herpes virus can be derived from different types of HSV, particularly preferred are
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`HSV1 and HSV2. The herpes virus may be genetically modified so as to restrict viral replication in
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`tumors or reduce its cytotoxicity in non—dividing cells. For example, any viral gene involved in
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`nucleic acid metabolism may be inactivated, such as thymidine kinase (Martuza et al., 1991, Science
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`252: 854—6), ribonucleotide reductase (RR) (Boviatsis et al., Gene Ther. 1: 323—31; Mineta et al.,
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`1994, Cancer Res. 54: 3363—66), or uracil—N—glycosylase (Pyles et al., 1994, J. Virol. 68: 4963—72).
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`Another aspect involves viral mutants with defects in the function of genes encoding virulence
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`factors such as the ICP34.5 gene (Chambers et al., 1995, Proc. Natl. Acad. Sci. USA 92: 1411-5).
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`Representative examples of oncolytic herpes virus include NV1020 (e.g. Geevarghese et al., 2010,
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`Hum. Gene Ther. 21(9): 1119-28) and T-VEC (Andtbacka et al., 2013, J. Clin. Oncol. 31, abstract
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`number LBA9008).
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`In one embodiment,
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`the oncolytic virus of the present
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`invention is obtained from a
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`morbillivirus which can be obtained from the paramyxoviridae family, with a specific preference for
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`measles virus. Representative examples of oncolytic measles viruses include without limitation MV-
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`Edm (McDonald et al., 2006; Breast Cancer Treat. 99(2): 177-84) and HMWMAA (Kaufmann et al.,
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`2013, J. Invest. Dermatol. 133(4): 1034-42)
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`In one embodiment, the oncolytic virus of the present invention is obtained from an
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`adenovirus. Methods are available in the art to engineer oncolytic adenoviruses. An advantageous
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`strategy includes
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`the replacement of viral promoters with tumor-selective promoters or
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`modifications of the E1 adenoviral gene product(s) to inactivate its/their binding function with p53
`
`or retinoblastoma (Rb) protein that are altered in tumor cells. In the natural context, the adenovirus
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`ElBSSkDa gene cooperates with another adenoviral product to inactivate p53 (p53 is frequently
`
`dysregulated in cancer cells),
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`thus preventing apoptosis. Representative examples of oncolytic
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`adenovirus include ONYX-015 (e.g. Khuri et al., 2000, Nat. Med 6(8): 879-85) and H101 also named
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`Oncorine (Xia et al., 2004, Ai Zheng 23(12): 1666-70).
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`In one embodiment, the oncolytic virus of the present invention is a poxvirus. As used
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`herein the term "poxvirus" refers to a virus belonging to the Poxviridae family, with a specific
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`preference for a poxvirus belonging to the Chordopoxviridae subfamily and more preferably to the
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`Orthopoxvirus genus. Sequences of the genome of various poxviruses, for example, the vaccinia
`
`virus, cowpox virus, Canarypox virus, Ectromelia virus, Myxoma virus genomes are available in the
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`art and specialized databases such as Genbank (accession number NC_006998, NC_003663,
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`NC_005309, NC_004105, NC_001132 respectively).
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`Desirably, the oncolytic poxvirus is an oncolytic vaccinia virus. Vaccinia viruses are members
`
`of the poxvirus family characterized by a 200kb double-stranded DNA genome that encodes
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`numerous viral enzymes and factors that enable the virus to replicate independently from the host
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`cell machinery. The majority of vaccinia virus particles is intracellular (IMV for intracellular mature
`
`virion) with a single lipid envelop and remains in the cytosol of infected cells until lysis. The other
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`infectious form is a double enveloped particle (EEV for extracellular enveloped virion) that buds out
`
`from the infected cell without lysing it.
`
`Although it can derive from any vaccinia virus strain, Elstree, Wyeth, Copenhagen and
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`Western Reserve strains are particularly preferred. The gene nomenclature used herein is that of
`
`Copenhagen vaccinia strain.
`
`It
`
`is also used herein for the homologous genes of other poxviridae
`
`unless otherwise indicated. However, gene nomenclature may be different according to the pox
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`strain but correspondence between Copenhagen and other vaccinia strains are generally available
`
`in the literature.
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`Preferably, the oncolytic vaccinia virus of the present invention is modified by altering for
`
`one or more viral gene(s). Said modification(s) preferably lead(s) to the synthesis of a defective
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`protein unable to ensure the activity of the protein produced under normal conditions by the
`
`unmodified gene (or
`
`lack of synthesis). Modifications encompass deletion, mutation and/or
`
`substitution of one or more nucleotide(s) (contiguous or not) within the viral gene or its regulatory
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`elements. Modification(s) can be made in a number of ways known to those skilled in the art using
`
`conventional recombinant techniques. Exemplary modifications are disclosed in the literature with
`
`a specific preference for those altering viral genes involved in DNA metabolism, host virulence, IFN
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`pathway (see e.g. Guse et al., 2011, Expert Opinion Biol.