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(12) Patent Application Publication (10) Pub. No.: US 2015/0023954 A1 Wu Et Al US 2015 0023954A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0023954 A1 Wu et al. (43) Pub. Date: Jan. 22, 2015 (54) POTENTIATING ANTIBODY-INDUCED GOIN33/574 (2006.01) COMPLEMENT-MEDIATED CYTOTOXCITY A613 L/4745 (2006.01) VAPI3K INHIBITION A613 L/4375 (2006.01) (52) U.S. Cl. (71) Applicant: MEMORIAL SLOAN-KETTERING CPC ....... A6IK39/39558 (2013.01); A61 K3I/4745 CANCER CENTER, New York, NY (2013.01); A61 K39/00II (2013.01); A61 K (US) 39/385 (2013.01); A61K31/4375 (2013.01); A6IK3I/5377 (2013.01); A61K.39/39 (72) Inventors: Xiaohong Wu, Forest Hills, NY (US); (2013.01); GOIN33/57484 (2013.01); CI2O Wolfgang W. Scholz, San Diego, CA I/6886 (2013.01); A61 K 2039/505 (2013.01); (US); Govind Ragupathi, New York, C12O 2600/16 (2013.01); C12O 2600/106 NY (US); Philip O. Livingston, (2013.01); A61K 2039/545 (2013.01) Bluffton, SC (US) USPC .................. 424/133.1; 424/277.1; 424/193.1; 424f1 74.1436/5O1435/7.1 : 506/9:435/7.92: (21) Appl. No.: 14/387,153 s s 435/6.12. 436/94 (22) PCT Filed: Mar. 14, 2013 (57) ABSTRACT (86). PCT No.: PCT/US 13/31278 Methodologies and technologies for potentiating antibody S371 (c)(1), based cancer treatments by increasing complement-mediated (2) Date: Sep. 22, 2014 cell cytotoxicity are disclosed. Further provided are method O O ologies and technologies for overcoming ineffective treat Related U.S. Application Data ments correlated with and/or caused by sub-lytic levels of (60) Provisional application No. 61/614,942, filed on Mar. complement-activating monoclonal antibodies (“mab') 23, 2012. against cancer antigens or cancer antigens with low tumor cell density. While detectable levels of passively administered or Publication Classification vaccine-induced mAb against Some antigens are able to delay or prevent tumor growth, low levels of mAb induce sublytic (51) Int. Cl. levels of complementactivation and accelerate tumor growth. A 6LX39/395 (2006.01) This complement-mediated accelerated tumor growth initi A6 IK39/00 (2006.01) ated by low mab levels results inactivation of the PI3K/AKT A 6LX39/385 (2006.01) Survival pathway. Methodologies and technologies relating to CI2O I/68 (2006.01) administration of PI3K inhibitors to overcome low dose A 6LX3/5377 (2006.01) mAb-initiated, complement-mediated PI3K activation and A 6LX39/39 (2006.01) accelerated tumor growth are disclosed. Patent Application Publication Jan. 22, 2015 Sheet 1 of 11 US 2015/0023954 A1 £38 --&isit ^ Ssss S$:S S 8 Patent Application Publication Jan. 22, 2015 Sheet 3 of 11 US 2015/0023954 A1 w C.A.33s. Nob sta 3 Sir E. a3. 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Sigire: S 88.388 % :%) US 2015/0023954 A1 Jan. 22, 2015 POTENTIATING ANTIBODY-INDUCED 0005 Complement-activating monoclonal antibodies COMPLEMENT-MEDIATED CYTOTOXCITY have been extensively utilized for the treatment of patients VAPI3K INHIBITION with tumors of different histotypes. Nonetheless, the overall importance of complement activation to the efficacy of mAb CROSS-REFERENCE TO RELATED based cancer therapies remains under investigation. Clini APPLICATIONS cally approved mouse anti-epithelial cell adhesion molecule and humanized anti-CD54 activate complement in vitro and 0001. This application claims priority to U.S. provisional medicate ADCC. m.Abs directed against HER2 and epithelial application Ser. No. 61/614,942, filed Mar. 23, 2012, the growth factor receptor 1 also activate complement in vitro. entirety of which is hereby incorporated herein by reference. Chimeric and mousemab against CD20 mediate tumoricidal effects in vivo through both ADCC and CDC. However, the BACKGROUND primary mechanism of action of other anti-tumor mAbs does not appear to involve complement. 0002 Monoclonal antibodies (“mab) are widely used in 0006. It has been postulated that the lytic potential of cancer therapy. They are utilized in a variety of ways, includ complementactivation by anti-cancer mAbs may be inhibited ing diagnosis, monitoring, and treatment of disease. When by membrane-bound complement regulatory proteins used therapeutically, monoclonal antibodies achieve their (mCRP). The level of complement activation on cell mem effects through various mechanisms. For example, some branes is regulated by the expression of mCRP, which block growth factor receptors, effectively arresting prolifera evolved to protect normal cells from uncontrolled comple tion of tumor cells. Alternatively or additionally, Some mono ment-mediated injury. mCRP comprise complement receptor clonal antibodies recruit cytotoxic effector cells such as 1 (CD35), membrane cofactor protein (CD46), decay-accel monocytes and macrophages through a process known as erating factor (CD55), and homologous restriction factor 20 antibody-dependent cell mediated cytotoxicity (ADCC). (CD59). CD35, CD46, and CD55 inhibit the deposition of C3 Some monoclonal antibodies bind complement, leading to fragments on the cell Surface and thereby limit complement direct cell death in a process known as complement depen dependent cellular cytotoxicity. CD59 prevents the formation dent cytotoxicity (“CDC). of membrane attack complexes and the Subsequent osmotic 0003. The complement system is an enzyme cascade com lysis of the target cell. Over-expression of these mCRP on prising a collection of blood and cell Surface proteins that tumor cells may prevent efficient complement-activation by assistantibodies in clearing pathogens from an organism. The anti-cancer antibodies. complement system comprises approximately 30 different proteins, including serum proteins, serosal proteins, and cell SUMMARY membrane receptors. Some complement proteins bind to 0007 Embodiments of the invention result from the sur immunoglobulins or to membrane components of cells. Oth prising discovery that while high levels of anti-tumor anti ers are proenzymes that, when activated, cleave one or more bodies have the ability to activate the complement cascade, other complement proteins and initiate an amplifying cascade low levels of anti-tumor antibodies can, in fact, induce Sub of further cleavages. The end-result of this cascade is massive lytic levels of complement activation and accelerate tumor amplification of the response and activation of the cell-killing growth. For example, although an anti-tumor, complement membrane attack complex. The complement system has four activating mAb may be administered at a sufficient dose to major functions, including lysis of infectious organisms, acti initially cause CDC or ADCC of the targeted cancer cell, in Vation of inflammation, opSonization and immune clearance. vivo levels of the mAb decrease over time. Thus, ironically, a 0004 Three different complement pathways have been therapeutically effective dose will eventually result in a low defined: the classical complement pathway, the alternative dose capable of propagating Survival and growth of remaining complement pathway, and the mannose-binding lectin path cells (i.e., Sublytic complement activation). This counter-in way. The classical pathway is activated following binding of tuitive high/low dichotomy is mediated by the phosphatidyli monoclonal antibodies (“mabs') to tumor cells. It is initiated nositol 3-kinase (“PI3K) cell survival pathway. The present by binding of the C1 complex to mabs in close proximity to invention further discloses that pharmacological inhibition of the tumor cell membranes. Complementactivation on the cell the PI3K pathway sensitizes cells to CDC mediated by anti surface results
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