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WO 2018/098352 A2 O (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2018/098352 A2 31 May 2018 (31.05.2018) W !P O PCT (51) International Patent Classification: (74) Agent: MURPHY, Amanda, K. et al; Finnegan, Hender 59/595 (2006.01) A61K 31/343 {2006.01) son, Farabow, Garrett & Dunner, LLP, 901 New York Av C07K 16/28 (2006.01) A61K 31/427 {2006.01) enue, NW, Washington, DC 20001-4413 (US). C12N 15/113 (2010.01) A61K 31/437 {2006.01) (81) Designated States (unless otherwise indicated, for every A61K 45/06 (2006.01) A61K 31/713 (2006.01) kind of national protection available): AE, AG, AL, AM, A61K 31/277 (2006.01) A61P 35/00 (2006.01) AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, A61K 31/337 (2006.01) CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, (21) International Application Number: DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, PCT/US2017/0631 16 HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, (22) International Filing Date: MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, 22 November 201 7 (22. 11.201 7) OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, (25) Filing Language: English 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. (26) Publication Language: English (84) Designated States (unless otherwise indicated, for every (30) Priority Data: kind of regional protection available): ARIPO (BW, GH, 62/425,358 22 November 20 16 (22. 11.20 16) US GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, (72) Inventors; and UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, (71) Applicants: OISHI, Jun [JP/JP]; c/o Sumitomo Dainippon TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, Pharma Co., Ltd., 1-91-98, Kasugadenaka 3-chome, Kono- EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, hana-ku, Osaka-shi, Osaka, 554-0022 (JP). SUN, Xiangao MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, [US/US]; 121 Bonad Road, Chestnut Hill, MA 02467 (US). TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, LI, Chiang, J. [US/US]; 8 Museum Way, Cambridge, MA KM, ML, MR, NE, SN, TD, TG). 02141 (US). (54) Title: TARGETING KRAS INDUCED IMMUNE CHECKPOINT EXPRESSION (57) Abstract: The interaction of PD-Ll expressed on the surface of tu mor cells triggers PD-1 signaling in T cells which contributes to resistance of cancers to emerging immune checkpoint therapies. The present disclo sure demonstrates that KRAS-induced RAF/MEK/ERK/FRA signaling is required for PD-Ll gene expression in tumor cells. Disclosed herein are compositions for the treatment of cancer comprising modulators of KRAS signaling combined with an immunotherapeutic agent that restores the sen sitivity of resistant tumor cells to the immunotherapeutic agent. For ex ample, a combination of a KRAS-specific asymmetric interfering RNAs (aiRNAs) and an immune checkpoint inhibitor is shown to enhance tumor cell-specific T cell cytotoxicity. Methods are also described for increasing the therapeutic efficacy of existing anticancer treatments through the co administration of a modulator of KRAS signaling. < © 00 FIG. 8E o o [Continued on next page] WO 2018/098352 A2 llll II II 11III II I II I II III 111 il II I II Published: — without international search report and to be republished upon receipt of that report (Rule 48.2(g)) TARGET G KRAS INDUCED IMMUNE CHECKPOINT EXPRESSION Together with natural killer cells and dendritic cells (DCs), cytotoxic T lymphocytes (CTLs) can orchestrate potent anti-tumor immune responses involving both adaptive and i nate effector mechanisms. Nonetheless, tumor cells are often able to evade this immune surveillance by commandeering immune checkpoint inhibitory pathways tha are hardwired into the immune system to retain self-tolerance and modulate the duration and amplitude of physiological immune responses in order to minimize potential collateral tissue damage. The PD-1 immune checkpoint pathway is one such example of an immune checkpoint that has emerged as a critical mediator of immunosuppression in the local tumor microenvironment. The inliibitory co-receptor Programmed Death 1 (PD-1; also known as CD279), a member of the extended CD28/CTLA-4 family of T cell regulators, is expressed on immune cells, such as T, B and NIC cells, whereas its ligand, the Programmed Cell Death Ligand 1 (PD-L1, also known as CD274 or B7-H1) is ce l surface glycoprotein expressed on the surface of tumor cells of solid tumors as well as on human tumor associated antigen presenting cells (APCs), e.g., dendritic cells and macrophages. The interaction of PD-L 1 ligand on tumor cells with the PD-1 receptor on immune cells delivers an inliibitory signal to T lymphocytes that ultimately leads to T cell anergy and immune evasion. The development of immune checkpoint inhibitors that prevent the activation of the PD-L 1/PD- immune checkpoint pathway has resulted in unprecedented and prolonged disease control in about 20-30% of cancer patients with melanoma, non-small cel lung cancer, renal cancer, or head/neck cancer (reviewed by Lipson et ah, Scmin. Oncol. (2015) 42(4): 87-600; Zou et at, Sci. Transl. Med. (2016) vol. 8, issue 328, pp. 328rv4). However, it remains unclear why the remaining 70-80% of cancer patients fail to respond to anti-PD-1 or anti-PD-Ll antibodies or why most patients with colorectal cancer, pancreatic cancer and other non- responsive tumor types are resistant to immune checkpoint inhibitors. Hence, there is a need in the art for treatment modalities targeting cancer ceils that are resistant or have acquired resistance to immune checkpoint therapies. The present disclosure is based on the discovery that aberrant KRAS signaling is at least in part responsible for the activation of PD-L 1 gene expression in tumor ceils and for the subsequent suppression of tumor cell-specific T cell toxicity. The present disclosure provides compositions and methods that can prevent tumors from evading immune surveillance through Lhe b activation of the PD- PD- immune checkpoint pathway in T cells in certain embodiments, inhibition of abeiTant KRAS signaling in tumor cells sensitizes tumor cells to immune checkpoint inhibitors. I certain embodiments, the disclosure further provides methods for enhancing the therapeutic efficacy of existing anticancer treatment using KRAS signaling modulators. In one aspect, a composition comprising an effective amount of a modulator of KRAS signaling is disclosed, wherein the modulator of KRAS signaling is effective at enhancing the sensitivity of a tumor cell to tumor cell-specific T cell cytotoxicity. In certain embodiments, the tumor cell is in a subj ect in certain embodiments, the modulator of KRAS signaling enhances the efficacy of a therapeutic agent at treating a KRAS associated disease, e.g. cancer. n certain embodiments, the modulator of KRAS signaling can be, for example, an inhibitor of aberrant KRAS signaling. In certain embodiments, the abeiTant KRAS signaling is induced by a modified KRAS, e.g., an oncogenic KRAS, expressed in tumor cells. In certai n embodiments, the aberrant KRAS signaling comprises the KRAS induced activation of at least one member of the RAS/R AF/ME 'ERK/F ~1 signal transduction pathway n certain embodiments, the abeiTant KRAS signaling can result irs the KRAS induced activation of PD- gene expression in tumor cells. ITS certain embodiments, the aberrant. KRAS signal ng cars be induced by an effector of KRAS signaling, e.g. by a KRAS GEF. In a second aspect, a composition is disclosed that comprises a combination of an effective amount of a modulator of KRAS signaling, and an effective amount of a therapeutic agent, wherein the modulator of KRAS signaling is effective at enhancing the sensitivity of a tumor cell to tumor cell-specific T cell cytotoxicity. In certain embodiments, the modulator of KRAS signaling ca be an inhibitor ofKRAS signaling. In certain embodiments, the modulator of KRAS signaling can enhance the efficacy of the therapeutic agent at treating a KRAS associated disease, for example, cancer. In certain embodiments, the tumor cell is in a subj ect. In a third aspect, a composition is disclosed that comprises a combination of an effective amount of an modulator of oncogenic KRAS signaling, and an effective amount of a therapeutic agent, wherein the modulator of oncogeni c KRAS signaling is effective at enhancing the sensitivity of a tumor cell to tumor cell-specific T cell cytotoxicity. In certain embodiments, the modulator of oncogenic KRAS signaling can be an inhibitor of oncogenic KRAS signaling. In certain embodiments, the modulator of oncogenic KRAS signaling can enhance e efficacy of the therapeutic agent at treating an oncogenic KRAS associated disease n certain embodiments, the tumor cell is in a subject. In certain embodiments, the therapeutic agent can be, for example, an anticancer therapeutic agent. I certain embodiments, the anticancer therapeutic agent can be, for example, an immunotherapeutic agent, such as an antigen-binding protein, or fragment thereof, e.g. an antibody that targets a cell surface antigen or extracellular growth factor. In certain embodiments, the anticancer therapeutic agent can be, for example, a small molecule inhibitor of a target protein required for the maintenance or progression of a cancer.
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