Combination therapy of type II anti-CD20 antibody and selective Bcl-2 inhibitor

Abstract

Combination therapies involving type II anti-CD20 antibodies and selective Bcl-2 inhibitors for treating patients suffering from cancer, particularly CD20-expressing cancers, are provided. [Solution] A method for treating cancer comprising administering an effective amount of GA101 antibody, or 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof for one or more administration periods.

Description

[Technical Field]

[0001] The present invention relates to combination therapy involving a type II anti-CD20 antibody and a selective Bcl-2 inhibitor for the treatment of patients suffering from cancer, particularly CD20-expressing cancers. [Background technology]

[0002] The CD20 molecule (also called human B-lymphocyte-restricted differentiation antigen or Bp35) is a hydrophobic transmembrane protein with a molecular weight of approximately 35 kD located on pre-B and mature B lymphocytes (Valentine, MA et al., J. Biol. Chem., 264(19)(1989)11282-11287; and Einfield, DA et al., EMBO J., 7(3)(1988)711-717). CD20 is present on the surface of more than 90% of B cells derived from peripheral blood or lymphoid organs, where it is expressed during early pre-B cell development and remains until plasma cell differentiation. CD20 is present on both normal and malignant B cells. In particular, CD20 is expressed on more than 90% of B-cell non-Hodgkin's lymphomas (NHL) (Anderson, KC et al., Blood, 63(6)(1984)1424-1433), but is not present on hematopoietic stem cells, pro-B cells, normal plasma cells, or other normal tissues (Tedder, TF et al., J. Immunol., 135(2)(1985)973-979).

[0003] The 85-amino acid carboxy-terminal region of the CD20 protein is located within the cytoplasm. The length of this region contrasts with that of other B cell-specific surface structures, such as IgM, IgD, and IgG heavy chains, or histocompatibility antigen class I1a or β cells, which have relatively short cytoplasmic regions of 3, 3, 28, 15, and 16 amino acids, respectively (Komaromy, M. et al., NAR, 11 (1983) 6775-6785). Of the last 61 carboxy-terminal amino acids, 21 are acidic residues, while only two are basic, indicating that this region has a strong net negative charge. The GenBank accession number is NP-690605. CD20 is involved in regulating early stages of B cell activation and differentiation processes (Tedder, TF et al., Eur. J. Immunol., 16 (1986) 881-887), and is thought to function as a calcium ion channel (Tedder, TF et al., J. Cell. Biochem., 14D (1990) 195).

[0004] There are two distinct types of anti-CD20 antibodies that differ significantly in their mode of CD20 binding and biological activity (Cragg, MS et al., Blood, 103 (2004) 2738-2743; and Cragg, MS et al., Blood, 101 (2003) 1045-1052). Type I antibodies, such as rituximab, are potent in complement-mediated cytotoxicity, whereas type II antibodies, such as tositumomab (B1), 11B8, AT80, or humanized B-Ly1 antibodies, effectively initiate target cell death via caspase-independent apoptosis accompanied by phosphatidylserine exposure.

[0005] The general characteristics shared by type I and type II anti-CD20 antibodies are summarized in Table 1 below.

[0006] [Table 1]

[0007] The Bcl-2 family of proteins regulates programmed cell death triggered by developmental stimuli in response to multiple stress signals (Cory, S., and Adams, JM, Nature Reviews Cancer, 2 (2002) 647-656; Adams, Genes und Development, 17 (2003) 2481-2495; Danial, NN, and Korsmeyer, SJ, Cell, 116 (2004) 205-219). Cell survival is promoted by Bcl-2 itself and several of its relatives (Bcl-xL, Bcl-W, Mcl-1, and A1) that contain three or four conserved Bcl-2 homology (BH) domains, whereas apoptosis is driven by two other subfamilies. The initial signal for cell death is transmitted by a diverse group of BH3-only proteins, including Bad, Bid, Bim, Puma, and Noxa, which all share a small BH3-interacting domain (Huang and Strasser, Cell 11, 103 (2000) 839-842). However, the involvement of Bax or Bak multidomain proteins, which contain BH1-BH3 domains, is required for cell death (Cheng et al., Molecular Cell, 8 (2001) 705-711; Wei, MC et al., Science, 292 (2001) 727-730; Zong, WX et al., Genes and Development, 15, 148 (2001) 1-1486). When activated, these proteins can permeabilize the outer mitochondrial membrane and release pro-apoptotic factors (e.g., cytochrome C) that are required to activate caspases that destroy the cell (Wang, K., Genes and Development, 15 (2001) 2922-2933; (Adams, 2003); Green, D.R., and Kroemer, G., Science, 305 (2004) 626-629).

[0008] The interaction between these three members of the Bcl-2 family determines whether cells live or die. When BH3-only proteins are activated, for example, in response to DNA damage, they can bind to the grooves on pro-survival associates via their BH3 domains (Sattler et al., Science, 275 (1997) 983-986). However, how BH3-only and Bcl-2-like proteins control the activation of Bax and Bak is poorly understood (Adams, 2003). Much attention has been focused on Bax. This soluble monomeric protein (Hsu, YT et al., Journal of Biological Chemistry, 272 (1997) 13289-13834; Wolter, KG et al., Journal of Cell Biology, 139 (1997) 1281-92) usually has its membrane targeting domain inserted into the groove, presumably for its cytosolic localization (Nechushtan, A. et al., EMBO Journal, 18 (1999) 2330-2341; Suzuki et al., Cell, 103 (2000) 645-654; Schinzel, A. et al., J. Cell Biol, 164 (2004) 1021-1032). Although several unrelated peptides / proteins have been proposed to modulate Bax activity, as reviewed in Lucken-Ardjomande, S., and Martinou, JC, J. Cell Sci., 118 (2005) 473-483, the physiological relevance of this has not yet been established. Bax can also be activated by direct engagement with specific BH3-only proteins (Lucken-Ardjomande, S., and Martinou, JC, 2005), the best documented of which is tBid, a truncated form of Bid (Wei, M.C. et al., Genes und Development, 14 (2000) 2060-2071; Kuwana, T. et al., Cell, 111 (2002) 331-342; Roucou, X. et al., Biochemical Journal, 368 (2002) 915-921; Cartron, P.F. et al., Mol Cell, 16 (2004) 807-818).As reviewed elsewhere (Adams, 2003), the oldest model in which Bcl-2 directly interacts with Bax (Oltvai, ZN et al., Cell, 74 (1993) 609-619) has been called into question because Bax is Bcl-bound while cytosolic, and the interaction appears to be significantly dependent on the detergent used for cell lysis (Hsu, YT, and Youle, 1997). Nevertheless, it is well established that the BH3 region of Bax can mediate binding to Bcl-2 (Zha, H., and Reed, J., Journal of Biological Chemistry, 272 (1997) 31482-88; Wang, K. et al., Molecular and Cellular Biology, 18 (1998) 6083-6089), and that Bcl-2 prevents Bax oligomerization, even though no heterodimers have been detected (Mikhailov, V. et al., Journal of Biological Chemistry, 276 (2001) 18361-18374). Thus, it remains uncertain whether pro-survival proteins directly or indirectly limit Bax activation.

[0009] Although Bax and Bak appear to be functionally equivalent in many environments (Lindsten, T. et al., Molecular Cell, 6 (2000) 1389-1399; Wei, M.C. et al., 2001), substantial differences in their regulation are predicted by their distinct localization in healthy cells. Unlike Bax, which is largely cytoplasmic, Bak is located in complexes on the outer membrane of mitochondria and the endoplasmic reticulum in healthy cells (Wei, M.C. et al., 2000; Zong, W.X. et al., Journal of Cell Biology, 162 (2003) 59-69). Nevertheless, upon receiving a cytotoxic signal, both Bax and Bak undergo conformational changes, Bax translocates to organelle membranes, and Bax and Bak then associate to form homo-oligomers that can lead to membrane permeabilization (Hsu, YT et al., PNAS, 94 (1997) 3668-3672; Wolter, KG et al., Uehara, 1997; Antonsson, B. et al., Journal of Biological Chemistry, 276 (2001) 11615-11623; Nechushtan, A. et al., Journal of Cell Biology, 153 (2001) 1265-1276; Wei, MC et al., Uehara, 2001; Mikhailov, V. et al., Journal of Biological Chemistry, 278 (2003) 5367-5376).

[0010] There are various Bcl-2 inhibitors, all of which have the same property of inhibiting pro-survival members of the Bcl-2 family of proteins and are therefore promising candidates for treating cancer. Such Bcl-2 inhibitors include, for example, oblimersen, SPC-2996, RTA-402, gossypol, AT-101, obatoclax mesylate, A-371191, A-385358, A-438744, ABT-737, ABT-263, AT-101, BL-11, BL-193, GX-15-003, 2-methoxyantimycin A3, HA-14-1, KF-67544, purpurogallin, TP-TW-37, YC-137, and Z-24, as described, for example, in Zhai, D. et al., Cell Death and Differentiation, 13 (2006) 1419-1421.

[0011] Smith, MR et al., Molecular Cancer Therapeutics, 3(12) (2004) 1693-1699; and Ramanarayanan, J. et al., British Journal of Haematology, 127(5) (2004) 519-530, refer to the combination of a type I anti-CD20 antibody (rituximab) with an antisense Bcl-2 oligonucleotide (oblimersen). [Prior art documents] [Non-patent literature]

[0012] [Non-Patent Document 1] Valentine, M.A. et al., J.Biol.Chem., 264(19)(1989)11282-11287 [Non-patent document 2] Einfield, DA et al., EMBO J., 7(3) (1988) 711-717 [Non-patent document 3] Anderson, KC et al., Blood, 63(6)(1984)1424-1433 [Non-patent document 4] Tedder,TFら,J,Immunol.,135(2)(1985)973-979

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[0013] The present invention provides a method for treating a patient suffering from cancer, comprising co-administering to a patient in need thereof a type II anti-CD20 antibody and a selective Bcl-2 inhibitor, which may be administered simultaneously or sequentially in any order.

[0014] An example of a type II anti-CD20 antibody for use in the present invention is the GA101 antibody.

[0015] In embodiments, the type II anti-CD20 antibody has high antibody-dependent cellular cytotoxicity (ADCC).

[0016] In embodiments, at least 40% of the oligosaccharides of the Fc region of the type II anti-CD20 antibody are non-fucosylated.

[0017] In embodiments, the selective Bcl-2 inhibitor is GDC-0199 (also known as ABT-199) or a pharmaceutically acceptable salt thereof.

[0018] In embodiments, the cancer is a non-solid tumor.

[0019] In one embodiment, a method is provided for treating cancer in a human in need thereof, comprising administering to the human multiple dosing cycles of GA101 antibody and / or GDC-0199. In embodiments, each dosing cycle of the multiple dosing cycles is at least 1 week. In embodiments, each dosing cycle of the multiple dosing cycles is at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks.

[0020] In an embodiment in which the GA101 antibody and GDC-0199 are administered to a human in multiple administration cycles, the GA101 antibody may be administered once per administration cycle during one or more of the multiple administration cycles. The amount of GA101 administered per administration may be, for example, in the range of about 300 mg to about 3000 mg, about 500 mg to about 3000 mg, or about 500 mg to about 1200 mg.

[0021] In embodiments in which the GA101 antibody and GDC-0199 are administered to a human in multiple administration cycles, GDC-0199 may be administered, for example, daily per administration cycle during one or more of the multiple administration cycles. In embodiments, GDC-0199 is administered for fewer than all days of an initial administration cycle, and daily during multiple administration cycles following the initial administration cycle. The amount of GDC-0199 administered per day may range from about 10 mg to about 1,000 mg, about 20 mg to about 800 mg, about 20 mg to about 500 mg, or about 50 mg to about 300 mg.

[0022] In embodiments, the GA101 antibody and GDC-0199 are administered to the patient in multiple administration cycles, at least 2, 3, 4, 5, 6, 7, 8, or more than 8 administration cycles.

[0023] In certain embodiments of the provided methods for treating cancer in a human in need thereof, comprising administering to the human multiple cycles after the final cycle of multiple cycles both a GA101 antibody and GDC-0199, only GDC-0199 may be administered to the human in the absence of the GA101 antibody, or only the GA101 antibody may be administered to the patient in the absence of GDC-0199. For example, when only GDC-0199 is administered to the human (e.g., after the final cycle of multiple cycles in which both GDC-0199 and the GA101 antibody are administered to the human), GDC-0199 may be administered to the human for at least 3, 4, 5, 6, 7, 8, or 9 days, or for 10 or more days, 20 or more days, or 30 or more days.

[0024] In yet another embodiment of the provided methods, in which the GA101 antibody and GDC-0199 are administered to the patient in multiple dosing cycles, the multiple dosing cycles include stepwise dosing cycles in which GDC-0199 is administered to the patient in escalating daily doses during the stepwise dosing cycles.

[0025] The present invention provides the combined use of GDC-0199 and GA101 antibody for the production of a medicament for the treatment of cancer. The present invention also provides the combined use of GA101 antibody and GDC-0199 for the production of a medicament for the treatment of cancer.

[0026] In another aspect, the present invention provides a combination of GA101 antibody and GDC-0199 for treating cancer in humans, which may be administered to humans, for example, according to the administration schedule described below. [Brief explanation of the drawings]

[0027] [Figure 1] Antitumor activity of combination therapy of a type II anti-CD20 antibody (GA101 antibody, in this case obinutuzumab) with a Bcl-2 inhibitor (ABT-199, also known as GDC-0199). Arrows and lines below the x-axis indicate the days of administration of GA101 and GDC-0199, respectively. [Figure 2]Exemplary dosing schedules for administering GDC-199 with obinutuzumab. [Figure 3] Exemplary dosing schedules for administering GDC-199 with obinutuzumab. [Figure 4] Antitumor activity of a type II anti-CD20 antibody (obinutuzumab, also known as RO5072759) used alone or together with GDC-0199, and a type I anti-CD20 antibody (rituximab) used alone or together with GDC-0199 against human Z138 mantle cell lymphoma cells. [Figure 5] Results from a xenograft model of aggressive lymphoma demonstrating that monotherapy with GDC-0199, following combination therapy with a type II anti-CD20 antibody (GA101 antibody, in this case obinutuzumab), delays tumor regrowth.

[0028] The present invention relates to the method described above.

[0029] The present invention provides a method for administering to a human in need thereof an effective amount of a GA101 antibody, or 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide, or a pharmaceutically acceptable salt thereof, for one or more dosing periods, followed by The present invention also relates to a method of treating a human comprising co-administering effective amounts of a GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide, or a pharmaceutically acceptable salt thereof, for one or more administration periods.

[0030] The present invention provides a method for administering to a human in need thereof an effective amount of GA101 antibody or 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide for 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days. and, after administration of the GA101 antibody, co-administering effective amounts of GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof for one or more administration periods.

[0031] The present invention also relates to a method for treating a human in need thereof, comprising administering an effective amount of a GA101 antibody for 1, 2, 3, 4, 5, 6, or 7 days, followed by co-administration of effective amounts of a GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof for one or more administration periods.

[0032] The present invention relates to a method for treating a GA101 antibody by administering an effective amount of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof for 1, 2, 3, 4, 5, 6, or 7 days, followed by administering an effective amount of GA101 antibody. and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide, or a pharmaceutically acceptable salt thereof, for one or more administration periods.

[0033] The present invention also relates to a method for treating a human in need thereof, comprising administering an effective amount of a GA101 antibody once per administration period for 1, 2, 3, 4, 5, or 6 cycles, followed by co-administration of an effective amount of a GA101 antibody once per administration period with 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof, 1 to 3 times daily during one or more administration periods.

[0034] The present invention relates to a method for treating a patient suffering from atopic dermatitis, comprising administering an effective amount of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof 1 to 3 times daily for 1, 2, 3, 4, 5, or 6 administration periods, followed by administering an effective amount of a GA101 antibody. and a pharmaceutically acceptable salt thereof, one to three times daily for one or more dosing periods.

[0035] The present invention relates to a method for treating rheumatoid arthritis, comprising administering to a subject a therapeutically effective amount of GA101 antibody, the effective amount of which is 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, or 3000 mg of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4 an effective amount of -((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof is 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 410, 420, 430, 440, 450, 460, 470, 480, 490, 510, 520, 530, 540, 550, 560, 570, 580, 590, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 0, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 11 20, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990 or 1000 mg.

[0036] The present invention relates to a method for treating a GA101 antibody, the effective amount of which is 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 mg, and a method for treating a GA101 antibody, the effective amount being ... The present invention also relates to any one of the above-described methods, wherein the effective amount of the 3-(4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof is 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, or 300 mg.

[0037] The present invention relates to a method for treating NHL, wherein the cancer is NHL, the effective amount of the GA101 antibody is 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 mg, and the effective amount of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof is 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 0, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 52 The present invention also relates to any one of the methods described above, wherein the amount of the active ingredient is 0, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, and 800 mg.

[0038] The present invention relates to a method for treating AML, wherein the cancer is AML, the effective amount of the GA101 antibody is 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 mg, and the effective amount of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof is 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 0, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 52 The present invention also relates to any one of the methods described above, wherein the amount of the active ingredient is 0, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, and 800 mg.

[0039] The present invention also relates to any one of the methods described above, wherein the GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof are co-administered sequentially during each administration period, and each administration period is 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days.

[0040] The term "antibody" is used broadly herein and encompasses a variety of antibody structures, including monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments as long as they exhibit the desired antigen-binding activity.

[0041] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies; i.e., except for possible variant antibodies containing naturally occurring mutations or arising during the production of the monoclonal antibody preparation, the individual antibodies comprising the population are identical and / or bind to the same epitope, with such variants usually being present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the modifier "monoclonal" refers to the character of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention can be produced by a variety of techniques, including, but not limited to, hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of the human immunoglobulin loci. These and other exemplary methods for producing monoclonal antibodies are described herein.

[0042] In one embodiment, the type II anti-CD20 antibody is a monoclonal antibody.

[0043] The term "chimeric antibody" refers to a monoclonal antibody, usually produced by recombinant DNA technology, that contains a variable region, i.e., a binding region, from one source or species and at least a portion of a constant region derived from a different source or species. Chimeric antibodies containing murine variable regions and human constant regions are particularly preferred. The aforementioned murine / human chimeric antibodies are the product of expressed immunoglobulin genes containing DNA segments encoding murine immunoglobulin variable regions and DNA segments encoding human immunoglobulin constant regions. Other forms of "chimeric antibodies" encompassed by the present invention are those in which the class or subclass has been modified or changed from that of the original antibody form. "Chimeric antibodies" are also referred to as "class-switched antibodies." Methods for producing chimeric antibodies include conventional recombinant DNA and gene transfection techniques known in the art. See, e.g., Morrison, SL et al., Proc. Natl. Acad. Sci. USA, 81 (1984) 6851-6855; U.S. Pat. No. 5,202,238; and U.S. Pat. No. 5,204,244.

[0044] The term "humanized antibody" refers to an antibody in which the framework or "complementarity-determining regions" (CDRs) have been modified to contain CDRs from an immunoglobulin with different specificity compared to the parent immunoglobulin. In a preferred embodiment, murine CDRs are grafted into the framework regions of a human antibody to create a "humanized antibody." See, e.g., Riechmann, L. et al., Nature, 332 (1988) 323-327; and Neuberger, MS et al., Nature, 314 (1985) 268-270. Particularly preferred CDRs correspond to those representing sequences recognizing the antigens described above for chimeric and bifunctional antibodies.

[0045] As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies are known in the art (van Dijk, MA, and van de Winkel, JG, Curr. Opin. Pharmacol., 5 (2001) 368-374). Based on this technology, human antibodies can be generated against a wide variety of targets. Examples of human antibodies are described, for example, in Kellermann, SA et al., Curr Opin Biotechnol., 13 (2002) 593-597.

[0046] As used herein, the term "recombinant human antibody" is intended to include all human antibodies that are produced, expressed, generated, or isolated by recombinant means, such as antibodies isolated from host cells (e.g., NSO or CHO cells) or animals (e.g., mice) transgenic for human immunoglobulin genes, or expressed using a recombinant expression vector transfected into a host cell. Recombinant human antibodies have variable and constant regions derived from rearranged human germline immunoglobulin sequences. Recombinant human antibodies according to the invention have undergone in vivo somatic hypermutation. Thus, the amino acid sequences of the VH and VL regions of the recombinant antibody are derived from and related to human germline VH and VL sequences, but may not naturally occur within the human antibody germline repertoire in vivo.

[0047] As used herein, "specifically binds" or "specifically binds to" refers to binding that is sufficiently selective for a target to be distinguished from binding to undesired or non-specific targets (e.g., antibodies that specifically bind to human CD20). In one embodiment, the GA101 antibody of the present invention binds to human CD20 at a binding affinity of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., ≦10 -8 M or less, e.g. 10 -8 M~10 -13 M, e.g. 10 -9 M~10-13 In yet another embodiment, the KD is 10 -10 mol / l or less (e.g., 10 -12 Binding affinity is determined by standard binding assays, such as Scatchard plot analysis on CD20-expressing cells.

[0048] As used herein, the term "nucleic acid molecule" is intended to include DNA molecules and RNA molecules. A nucleic acid molecule can be single-stranded or double-stranded. In one embodiment, the nucleic acid molecule is double-stranded DNA.

[0049] The "constant domains" are not involved directly in binding an antibody to an antigen, but are involved in effector functions (ADCC, complement fixation, and CDC).

[0050] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to an antigen. The variable domains of the heavy and light chains (VH and VL, respectively) of natural antibodies usually have similar structures, with each domain containing four conserved framework regions (FR) and three hypervariable regions (HVR) (see, e.g., Kindt et al., Kuby Immunology, 6 th ed., W.H. Freeman and Co., p. 91 (2007).

[0051] As used herein, the term "hypervariable region" or "HVR" refers to each of the regions of an antibody variable domain that are hypervariable in sequence ("complementarity determining regions" or "CDRs") and / or form structurally defined loops ("hypervariable loops") and / or contain antigen-bearing residues ("antigen contacts"). Typically, antibodies contain six HVRs: three HVRs in the VH (H1, H2, H3) and three HVRs in the VL (L1, L2, L3). Examples of HVRs in the present invention include: (a) hypervariable loops present at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol., 196:901-917 (1987)); (b) CDRs located at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991)); (c) antigenic contacts occurring at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al., J. Mol. Biol., 262:732-745 (1996)); and (d) combinations of (a), (b) and / or (c) including HVR amino acid residues 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3), and 94-102 (H3); Includes:

[0052] Art-recognized synonyms of CD20 include B lymphocyte antigen CD20, B lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5.

[0053] According to the present invention, the term "anti-CD20 antibody" refers to an antibody that specifically binds to the CD20 antigen. Depending on the binding properties and biological activity of the anti-CD20 antibody to the CD20 antigen, two types of anti-CD20 antibodies (type I and type II anti-CD20 antibodies) can be distinguished according to Cragg, MS et al., Blood, 103 (2004) 2738-2743; and Cragg, MS et al., Blood, 101 (2003) 1045-1052 (see Table 2).

[0054] [Table 2]

[0055] One characteristic of type I and type II anti-CD20 antibodies is their binding mode. Type I and type II anti-CD20 antibodies can be classified by the ratio of their binding ability to CD20 on Raji cells (ATCC-No. CCL-86) compared to rituximab.

[0056] The type II anti-CD20 antibody has a ratio of binding ability to CD20 on Raji cells (ATCC-No. CCL-86) of 0.3 to 0.6, in one embodiment 0.35 to 0.55, and in another embodiment 0.4 to 0.5, of the anti-CD20 antibody to rituximab. Examples of type II anti-CD20 antibodies include tositumomab (B1 IgG2a), GA101 antibody IgG1 (a chimeric humanized IgG1 antibody disclosed in WO 2005 / 044859), 11B8 IgG1 (disclosed in WO 2004 / 035607), and AT80 IgG1. In one embodiment, the type II anti-CD20 antibody (disclosed in WO 2005 / 044859) is a monoclonal antibody that binds to the same epitope as the GA101 antibody.

[0057] The "ratio of anti-CD20 antibody to rituximab in binding ability to CD20 on Raji cells (ATCC-No. CCL-86)" is determined by direct immunofluorescence measurement (mean fluorescence intensity (MFI) is measured) using Cy5-conjugated anti-CD20 antibody and Cy5-conjugated rituximab in a FACSArray (Becton Dickinson) using Raji cells (ATCC-No. CCL-86) as described in Example 2, and calculated as follows:

[0058]

number

[0059] MFI means mean fluorescence intensity. As used herein, "Cy5 labeling ratio" means the number of Cy5-labeled molecules per molecule antibody.

[0060] Typically, the type II anti-CD20 antibody has a ratio of the second anti-CD20 antibody to rituximab in binding ability to CD20 on Raji cells (ATCC-No. CCL-86) of 0.3 to 0.6, in one embodiment 0.35 to 0.55, and in yet another embodiment 0.4 to 0.5.

[0061] In one embodiment, the type II anti-CD20 antibody, for example, the GA101 antibody, has high antibody-dependent cellular cytotoxicity (ADCC).

[0062] By "antibody with high antibody-dependent cellular cytotoxicity (ADCC)" is meant an antibody that has high ADCC as measured by any suitable method known to those skilled in the art, as the term antibody is defined herein. One recognized in vitro ADCC assay is as follows: 1) the assay uses target cells known to express the target antigen recognized by the antigen-binding region of the antibody; 2) the assay uses human peripheral blood mononuclear cells (PBMCs) isolated from the blood of randomly selected healthy donors as effector cells; 3) The assay is carried out according to the following protocol: i) PBMCs were isolated using standard density centrifugation procedures and cultured at 5 x 10 cells in RPMI cell culture medium. 6 Suspend at cells / ml; ii) Target cells are grown by standard tissue culture methods, harvested from exponential growth phase with greater than 90% viability, washed with RPMI cell culture medium, and diluted with 100 microcuries of 51 Label with Cr, wash twice with cell culture medium, and then add 10 ml of cell culture medium. 5 Resuspend at a density of cells / ml; iii) transferring 100 μl of the final target cell suspension to each well of a 96-well microtiter plate; iv) serially diluting the antibody in cell culture medium from 4000 ng / ml to 0.04 ng / ml and adding 50 μl of the resulting antibody solution to target cells in a 96-well microtiter plate, testing in triplicate at various antibody concentrations covering the entire concentration range described above; v) For maximum release (MR) controls, three additional wells in the plate containing labeled target cells receive 50 μl of a 2% (VN) solution of non-ionic detergent (Nonidet, Sigma, St. Louis) in water instead of the antibody solution (point iv above); vi) For spontaneous release (SR) controls, three additional wells in the plate containing labeled target cells receive 50 μl of RPMI cell medium instead of the antibody solution (point iv above); vii) The 96-well microtiter plate is then centrifuged at 50×g for 1 minute and incubated at 4° C. for 1 hour; viii) Add 50 μl of PBMC suspension (point i above) to each well to yield an effector:target cell ratio of 25:1 and place the plate in an incubator at 37° C. in a 5% CO atmosphere for 4 hours; ix) collecting the cell-free supernatant from each well and quantifying the experimentally released radioactivity (ER) using a gamma counter; x) Calculate the percentage of specific lysis for each antibody concentration according to the formula (ER-MR) / (MR-SR) x 100, where ER is the mean radioactivity quantified for that antibody concentration (see point ix above), MR is the mean radioactivity quantified for the MR control (see point v above) (see point ix above), and SR is the mean radioactivity quantified for the SR control (see point vi above) (see point ix above). 4) "High ADCC" is defined as an increase in the maximum percentage of specific lysis observed within the antibody concentration range tested above, and / or a decrease in the concentration of antibody required to achieve half of the maximum percentage of specific lysis observed within the antibody concentration range tested above. In one embodiment, the increase in ADCC is relative to ADCC mediated by the same antibody and produced by the same type of host cell, as determined in the above assay using the same standard production, purification, formulation, and storage methods known to those of skill in the art, except that the comparator antibody (lacking high ADCC) was not produced from host cells engineered to overexpress GnTIII and / or engineered to have low expression from the fucosyltransferase 8 (FUT8) gene (including, for example, those engineered for FUT8 knockout).

[0063] "High ADCC" can be obtained, for example, by mutating and / or glycoengineering the antibody. In one embodiment, the antibody is glycoengineered to have biantennary oligosaccharides attached to the Fc region of the antibody, which is bisected by GlcNAc, e.g., as described in WO 2003 / 011878 (Jean-Mairet et al.); US Patent No. 6,602,684 (Umana et al.); US 2005 / 0123546 (Umana et al.); Umana, P. et al., Nature Biotechnol., 17 (1999) 176-180. In another embodiment, the antibody is glycoengineered to lack fructose on the carbohydrate attached to the Fc region by expressing the antibody in host cells that lack protein fucosylation (e.g., Lec13 CHO cells or cells with an α-1,6-fucosyltransferase gene (FUT8) deletion or FUT gene expression knockdown (see, e.g., Yamane-Ohnuki et al., Biotech. Bioeng., 87:614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO 2003 / 085107). In yet another embodiment, the antibody sequence is engineered to enhance ADCC in its Fc region (e.g., in one embodiment, the engineered antibody variant comprises an Fc region with one or more amino acid substitutions at positions 298, 333, and / or 334 (EU numbering of residues) of the Fc region).

[0064] The term "complement-dependent cytotoxicity (CDC)" refers to the lysis of human tumor target cells by an antibody according to the invention in the presence of complement. CDC can be determined by treating a preparation of CD20-expressing cells with an anti-CD20 antibody according to the invention in the presence of complement. CDC is observed if the antibody induces 20% or more lysis (cell death) of tumor cells after 4 hours at a concentration of 100 nM. In one embodiment, the assay is 51 This was performed using Cr or Eu labeled tumor cells. 51 Cr or Eu is measured. Controls include incubation of tumor target cells with complement but no antibody.

[0065] As used herein, the term "GA101 antibody" refers to any one of the following antibodies that bind to human CD20: (1) an antibody comprising an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 1, an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 2, an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 3, an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 4, an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 5, and an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 6; (2) an antibody comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 7 and a VL domain comprising the amino acid sequence of SEQ ID NO: 8; (3) an antibody comprising the amino acid sequence of SEQ ID NO: 9 and the amino acid sequence of SEQ ID NO: 10; (4) an antibody known as obinutuzumab; or (5) an antibody comprising an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 9 and an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 10. In one embodiment, the GA101 antibody is an IgG1 isotype antibody.

[0066] Oligosaccharide components can significantly influence properties related to the efficacy of therapeutic glycoproteins, such as physical stability, resistance to protease attack, interaction with the immune system, pharmacokinetics, and specific physiological activity. These properties may depend not only on the presence or absence of oligosaccharides, but also on the specific structure of the oligosaccharides. Some generalizations can be made between oligosaccharide structures and glycoproteins. For example, certain oligosaccharide structures mediate rapid clearance of glycoproteins from the bloodstream through interaction with specific carbohydrate-binding proteins, whereas other oligosaccharide structures may be bound by antibodies and trigger unwanted immune responses (Jenkins, N., et al., Nature Biotechnol., 14 (1996) 975-981).

[0067] Mammalian cells are preferred hosts for the production of therapeutic glycoproteins due to their ability to glycosylate proteins in a form most compatible with human applications (Cumming, DA, et al., Glycobiology, 1 (1991) 115-130; Jenkins, N., et al., Nature Biotechnol., 14 (1996) 975-981). Bacteria glycosylate proteins very rarely, and other similar host types, such as yeast, filamentous fungi, insect, and plant cells, produce glycosylation patterns associated with rapid clearance from the bloodstream, undesirable immune interactions, and, in some specific cases, low biological activity. Among mammalian cells, Chinese hamster ovary (CHO) cells have been the most commonly used for the last 20 years. In addition to providing appropriate glycosylation patterns, these cells routinely generate genetically stable, highly productive clonal cell lines. These can be cultured to high densities in simple bioreactors using serum-free media, allowing the development of safe and reproducible bioprocesses. Other commonly used animal cells include baby hamster kidney (BHK) cells, NSO- and SP2 / 0-mouse myeloma cells. More recently, production from transgenic animals has also been tested (Jenkins, N. et al., Nature Biotechnol., 14 (1996) 975-981).

[0068] All antibodies contain carbohydrate structures at conserved positions in the heavy chain constant region, and each isotype possesses a distinct array of N-linked carbohydrate structures that variably affect protein assembly, secretion, or functional activity (Wright, A., and Monison, S.L., Trends Biotech., 15 (1997) 26-32). The structures of the N-linked carbohydrates attached vary greatly depending on the degree of processing and can include high-mannose, multiantennary, and biantennary complex oligosaccharides (Wright, A., and Morrison, S.L., Trends Biotech., 15 (1997) 26-32). Typically, there is heterogeneous processing of the core oligosaccharide structure attached at a particular glycosylation site, such that monoclonal antibodies exist as multiple glycoforms. Similarly, it has been found that antibody glycosylation varies greatly between cell lines, with smaller variations observed within a given cell line grown under different culture conditions (Lifely, M.R. et al., Glycobiology, 5 (1995) 813-822).

[0069] One way to greatly increase efficacy while maintaining simple production methods and potentially avoiding significant undesirable side effects is to enhance the natural cell-mediated effector functions of monoclonal antibodies by engineering oligosaccharide moieties, as described in Umana, P., et al., Nature Biotechnol., 17 (1999) 176-180 and U.S. Pat. No. 6,602,684. IgG1-type antibodies, the most commonly used antibodies in cancer immunotherapy, are glycoproteins that have a conserved N-linked glycosylation site at Asn297 in each CH2 domain. Two complex-type biantennary oligosaccharides attached to Asn297 are buried between the CH2 domains, forming extensive contacts with the polypeptide backbone, and their presence is essential for antibodies to mediate effector functions such as antibody-dependent cellular cytotoxicity (ADCC) (Lifely, MR et al., Glycobiology, 5 (1995) 813-822; Jefferis, R. et al., Immunol. Rev., 163 (1998) 59-76; Wright, A., and Morrison, SL, Trends Biotechnol., 15 (1997) 26-32).

[0070] It has previously been shown that overexpression of β(1,4)-N-acetylglucosaminyltransferase III (GnTIII), a glycosyltransferase that catalyzes the formation of bisecting oligosaccharides, in Chinese hamster ovary (CHO) cells greatly increases the in vitro ADCC activity of an anti-neuroblastoma chimeric monoclonal antibody (chCE7) produced by the engineered CHO cells (see Umana, P. et al., Nature Biotechnol., 17 (1999) 176-180; and WO 99 / 154342, the entire contents of which are incorporated herein by reference). The antibody chCE7 belongs to a large class of unconjugated monoclonal antibodies that have high tumor affinity and specificity but are unlikely to be clinically useful when produced in standard industrial cell lines that lack the GnTIII enzyme (Umana, P. et al., Nature Biotechnol., 17 (1999) 176-180). This work first demonstrated that a large increase in ADCC activity can be obtained by engineering antibody-producing cells to express GnTIII, increasing the proportion of bisected oligosaccharides associated with the constant region (Fc), including bisected nonfucosylated oligosaccharides, above the levels found in naturally occurring antibodies.

[0071] In one embodiment, a composition comprising the GA101 antibody of the present invention comprises a GA101 antibody that has been engineered to have high ADCC activity.

[0072] As used herein, the term "Bcl-2" refers to the Bcl-2 protein (Swiss Prot ID No. P10415), a member of the Bcl-2 family of proteins (Cory, S., and Adams, JM, Nature Reviews Cancer, 2 (2002) 647-656; Adams, Genes und Development, 17 (2003) 2481-2495; Danial, NN, and Korsmeyer, SJ, Cell, 116 (2004) 205-219; Petros, AM, Biochim Biophys Acta, 1644 (2004) 83-94).

[0073] As used herein, the term "selective Bcl-2 inhibitor" refers to the Bcl-2 inhibitor of Formula I, 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide (also known as ABT-199 or GDC-0199), which is described in International Publication WO 2010 / 138588 and U.S. Patent Publication No. US 2010 / 0305122, both of which are incorporated herein by reference.

[0074] [ka]

[0075] The term "expression of CD20 antigen" is intended to refer to significant levels of expression of CD20 antigen on cells, e.g., T or B cells. In one embodiment, a patient to be treated according to the methods of the present invention expresses significant levels of CD20 on a B cell tumor or cancer. Patients with "CD20-expressing cancers" can be determined by standard assays known in the art. For example, CD20 antigen expression can be determined using immunohistochemical (IHC) detection, FACS, or by PCR-based detection of corresponding mRNA.

[0076] As used herein, the term "CD20-expressing cancer" refers to any cancer in which cancer cells express the CD20 antigen. CD20-expressing cancers include, for example, lymphoma, lymphocytic leukemia, lung cancer, non-small cell lung (NSCL) cancer, bronchioloalveolar cell lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, colon cancer, breast cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, and the like. The cancer may be a sarcoma of soft tissue, urethral cancer, penile cancer, prostate cancer, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic cancer, mesothelioma, hepatocellular carcinoma, biliary tract cancer, central nervous system (CNS) tumor, tumor of the spinal axis, brain stem glioma, glioblastoma multiforme, astrocytoma, schwannoma, ependymoma, medulloblastoma, meningioma, squamous cell carcinoma, pituitary adenoma, including refractory forms of any of the above cancers, or a combination of one or more of the above cancers.

[0077] In one embodiment, CD20-expressing cancer as used herein refers to lymphomas (e.g., B-cell non-Hodgkin's lymphoma (NHL)) and lymphocytic leukemias. Lymphomas and lymphocytic leukemias include, among other types of lymphomas and lymphocytic leukemias, for example: a) follicular lymphoma; b) small non-cleaved cell lymphoma / Burkitt lymphoma (including endemic Burkitt lymphoma, sporadic Burkitt lymphoma, and non-Burkitt lymphoma); c) marginal zone lymphoma (including extranodal marginal zone B-cell lymphoma (mucosa-associated lymphoid tissue lymphoma, MALT), nodal marginal zone B-cell lymphoma, and splenic marginal zone lymphoma); d) mantle cell lymphoma (MCL); e) large cell lymphoma (including B-cell diffuse large cell lymphoma ( DLCL), diffuse mixed cell lymphoma, immunoblastic lymphoma, primary mediastinal B-cell lymphoma, angiocentric lymphoma - including pulmonary B-cell lymphoma), f) hairy cell leukemia, g) lymphocytic lymphoma, Waldenström's macroglobulinemia, h) acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL) / small lymphocytic lymphoma (SLL), B-cell prolymphocytic leukemia, i) plasma cell neoplasms, plasma cell myeloma, multiple myeloma, plasmacytoma, j) Hodgkin's disease, k) acute myeloid leukemia (AML).

[0078] In one embodiment, the CD20-expressing cancer is B-cell non-Hodgkin's lymphoma (NHL). In another embodiment, the CD20-expressing cancer is mantle cell lymphoma (MCL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), B-cell diffuse large cell lymphoma (DLCL), acute myeloid leukemia (AML), Burkitt's lymphoma, hairy cell leukemia, follicular lymphoma, multiple myeloma, marginal zone lymphoma, post-transplant lymphoproliferative disorder (PTLD), HIV-associated lymphoma, Waldenstrom's macroglobulinemia, or primary CNS lymphoma.

[0079] As used herein, "relapsed or refractory" CLL includes CLL patients who have received at least one prior chemotherapy-containing treatment regimen. Relapsed patients usually have progressive disease that develops after responding to a prior chemotherapy-containing treatment regimen. Refractory patients usually have failed to respond to or relapsed within six months of the last prior chemotherapy-containing regimen.

[0080] As used herein, "untreated" CLL includes patients who have been diagnosed with CLL but have not typically received prior chemotherapy or immunotherapy. Patients with a history of emergency, locoregional radiation therapy (e.g., to relieve pressure signs or symptoms), or corticosteroids may be considered to have not yet received treatment.

[0081] The term "treating," as used herein, unless otherwise indicated, means partially or completely reversing, mitigating, inhibiting, or partially or completely preventing tumor growth, tumor metastasis, or other oncogenic or neoplastic cell progression in a patient. The term "treatment," as used herein, unless otherwise indicated, refers to the act of treating.

[0082] For example, when applied to cancer, the term "treatment method" or its equivalents refers to a procedure or course of action intended to reduce or eliminate the number of cancer cells or alleviate the symptoms of cancer in a patient. A "treatment method" of cancer or another proliferative disorder does not necessarily mean that cancer cells or other disease are actually eliminated, that the number of cells or disease is actually reduced, or that the cancer or other disease is actually alleviated. Often, cancer treatment methods are performed despite a low probability of success, but are deemed to induce an overall beneficial course of action given the patient's medical history and estimated life expectancy. The term "co-administration" or "co-administering" refers to the administration of a type II anti-CD20 antibody and a selective Bcl-2 inhibitor as two separate formulations. Co-administration can be simultaneous or sequential in either order. In a further embodiment, there is a period during which both (or all) active agents simultaneously exert their biological activities. The type II anti-CD20 antibody and the selective Bcl-2 inhibitor are co-administered simultaneously or sequentially intravenously (i.v.), e.g., during continuous infusion (for the antibody and then for the Bcl-2 inhibitor; or the Bcl-2 inhibitor is administered orally). When both therapeutic agents are co-administered sequentially, the therapeutic agents are administered in two separate administrations separated by a "specified period of time." The term "specified period of time" means between 1 hour and 15 days. For example, one of the therapeutic agents may be administered about 15, 14, 13, 12, or 24 hours after the administration of the other therapeutic agent. , 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 day, or 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 hour, and in one embodiment the specified period is 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 day, or 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 hour.

[0083] The term "concurrently" means at the same time or within a short period of time, usually less than an hour.

[0084] As used herein, the administration period refers to the period during which each therapeutic agent is administered at least once. The administration cycle is usually about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 days, and in one embodiment, 6, 7, 8, 9, 10, 11, 12, 13 or 14 days, for example, 7 or 14 days.

[0085] In certain embodiments, the administration period is a dosing cycle.

[0086] It is understood that antibodies are administered to patients in a "therapeutically effective amount" (or simply "effective amount"), which is the amount of each compound or combination that elicits the biological or medical response in a tissue, system, animal, or human that is desired by a researcher, veterinarian, physician, or other clinician. An effective amount of a therapeutic agent can be administered as a single dose or in divided doses. "Split-dose administration" means that the effective amount is divided into multiple doses, preferably two, and administered within one or two days. For example, if 100 mg of a selective Bcl-2 inhibitor is deemed effective, one 100 mg dose or two 50 mg doses may be administered. It is sometimes desirable to administer a divided dose at the beginning of a dosing period to reduce side effects. Even when an effective dose is administered in divided doses, a single administration of the effective amount is also contemplated. For example, if 100 mg is an effective amount of a selective Bcl-2 inhibitor and two 50 mg doses are administered over a period of time, e.g., two days, the effective amount is administered only once during that period.

[0087] The amount and timing of co-administration of a type II anti-CD20 antibody and a Bcl-2 inhibitor depend on the type (species, sex, age, weight, etc.) and condition of the patient being treated, as well as the severity of the disease or condition being treated. Suitably, the type II anti-CD20 antibody and the Bcl-2 inhibitor are co-administered to the patient at one time or over a series of treatments. Depending on the type and severity of the disease, initial candidate doses for co-administration of both drugs to a patient range from about 1 μg / kg to 50 mg / kg (e.g., 0.1 to 20 mg / kg) of a type II anti-CD20 antibody and 0.1 mg / kg to 200 mg / kg (e.g., 10 to 150 mg / kg) of a selective Bcl-2 inhibitor. If administered intravenously, the initial infusion time of the type II anti-CD20 antibody or Bcl-2 inhibitor may be longer than subsequent infusion times, e.g., about 90 minutes for the initial infusion and about 30 minutes for subsequent infusions (if the initial infusion is well tolerated).

[0088] In one embodiment, the preferred dose of the type II anti-CD20 antibody ranges from about 0.05 mg / kg to about 30 mg / kg, preferably from 1 mg / kg to 30 mg / kg, or is a flat dose of 500 mg to 3000 mg. Thus, one or more doses of about 0.5 mg / kg, 2.0 mg / kg, 4.0 mg / kg, 10 mg / kg, or 30 mg / kg, or a flat dose of 500 mg to 3000 mg (or a combination thereof) may be co-administered to the patient. The preferred dose of the Bcl-2 inhibitor ranges from 20 mg / kg to about 150 mg / kg, preferably from 1 mg / kg to 10 mg / kg. Depending on the type (species, sex, age, weight, etc.) and condition of the patient, as well as the type of anti-CD20 antibody and Bcl-2 inhibitor, the dose and administration schedule of the anti-CD20 antibody may differ from the dose of the Bcl-2 inhibitor. For example, the anti-CD20 antibody may be administered, for example, once every 1-3 weeks, and the Bcl-2 inhibitor may be administered daily or every 2-7 days. A high loading dose may be administered initially, followed by one or more lower doses.

[0089] The present invention relates, in part, to compositions comprising a type II anti-CD20 antibody and a selective Bcl-2 inhibitor.

[0090] In a preferred embodiment, the compositions of the invention are useful for preventing or inhibiting metastasis or further dissemination in patients suffering from CD20-expressing cancers. The compositions are useful for extending the survival of said patients, extending the progression-free survival of said patients, or extending the duration of response, resulting in a statistically significant and clinically meaningful improvement in treated patients as measured by survival, progression-free survival, response rate, or duration of response. In a preferred embodiment, the compositions are useful for enhancing the response of a patient population.

[0091] The present invention may involve the use of additional cytotoxic, chemotherapeutic, or anti-cancer agents or compounds that enhance the effects of the agents (e.g., cytokines) in combination therapy of type II anti-CD20 antibodies and Bcl-2 inhibitors for CD20-expressing cancers. Suitably, the molecules are present in the combination in amounts that are effective for the intended purpose. Preferably, the type II anti-CD20 antibody and Bcl-2 inhibitor combination therapy is used without additional cytotoxic, chemotherapeutic, or anti-cancer agents or compounds that enhance the effects of the agents.

[0092] Examples of such substances include alkylating agents having alkylating activity, such as cyclophosphamide (CTX; e.g., Cytoxan®), chlorambucil (CHL; e.g., Leukeran®), cisplatin (CisP; e.g., Platinol®), busulfan (e.g., Myleran®), melphalan, carmustine (BCNU), streptozotocin, triethylenemelamine (TEM), mitomycin C, and the like; and metabolic antagonists, such as methotrexate (MTX), etoposide (VP16; e.g., Bepesid®), 6-mercaptopurine (6MP), 6-thioguanine (6TG), cytarabine (Ara-C), 5-fluorouracil (5-FU), capecitabine (e.g., Xeloda®), and dacarbazine (DTIC). etc.; antibiotics such as actinomycin D, doxorubicin (DXR; e.g., Adriamycin®), daunorubicin (daunomycin), bleomycin, mithramycin, etc.; alkaloids such as vin alkaloids such as vincristine (VCR), vinblastine, etc.; and other antitumor agents such as paclitaxel (e.g., Taxol®) and paclitaxel derivatives, cytostatics, glucocorticoids (e.g., dexamethasone (DEX; e.g., Decadron®)) and corticosteroids (e.g., prednisone), nucleoside enzyme inhibitors (e.g., hydroxyurea), amino acid depleting enzymes (e.g., asparaginase), leucovorin and other folic acid derivatives, and various similar antitumor agents.Additional substances include amifostine (e.g., Ethiol®), dactinomycin, mechlorethamine (nitrogen mustard), streptozocin, cyclophosphamide, lomustine (CCNU), doxorubicin lipoprotein (e.g., Doxil®), gemcitabine (e.g., Gemzar®), daunorubicin lipoprotein (e.g., Daunoxome®), procarbazine, mitomycin, docetaxel (e.g., Taxotere®), aldesleukin, carboplatin, oxaliplatin, cladribine, camptothecin, and CPT. 11 (irinotecan), 10-hydroxy-7-ethyl-camptothecin (SN38), floxuridine, fludarabine, ifosfamide, idarubicin, mesna, interferon beta, interferon alpha, mitoxantrone, topotecan, leuprolide, megestrol, melphalan, mercaptopurine, plicamycin, mitotane, pegaspargase, pentostatin, pipobroman, plicamycin, tamoxifen, teniposide, testolactone, thioguanine, thiotepa, uracil mustard, vinorelbine, and chlorambucil may also be used. Preferably, a type II anti-CD20 antibody and Bcl-2 inhibitor combination therapy is used without the additional substances.

[0093] The use of the above-described cytotoxic and anti-cancer agents, as well as anti-proliferative target-specific anti-cancer agents, such as protein kinase inhibitors, in chemotherapy regimens is generally well characterized in the cancer treatment industry, and their use in the present invention is similarly considered, with some adjustments, to monitor tolerance and efficacy and to control the route and dose of administration. For example, the actual dose of a cytotoxic agent may vary depending on the patient's cultured cell response, as determined using tissue culture methods. Typically, the dose is reduced compared to the amount used in the absence of additional other substances.

[0094] Typical dosages of effective cytotoxic agents can be in the range recommended by the manufacturer, and can be reduced by up to about an order of magnitude in concentration or amount if one or more in vitro responses in animal models are demonstrated. Thus, the actual dosage will depend on the judgment of the physician, the condition of the patient, and the effectiveness of the therapeutic method based on the in vitro response of primary malignant cells or tissue cultured tissue samples or the response observed in an appropriate animal model.

[0095] In the present invention, in addition to type II anti-CD20 antibody and Bcl-2 inhibitor combination therapy for CD20-expressing cancers, an effective amount of ionizing radiation may be administered and / or a radiopharmaceutical may be used. The radiation source may be either external or internal to the patient being treated. When the radiation source is external to the patient, the treatment is known as external beam radiation therapy (EBRT). When the radiation source is internal to the patient, the treatment is called brachytherapy (BT). Radioactive atoms for use in the present invention may be selected from the group including, but not limited to, radium, cesium-137, iridium-192, americium-241, gold-198, cobalt-57, copper-67, technetium-99, iodine-123, iodine-131, and indium-111. It is also possible to label the antibody with a radioisotope. It is preferable to use type II anti-CD20 antibody and Bcl-2 inhibitor combination therapy without ionizing radiation.

[0096] Radiation therapy is the standard treatment for controlling unresectable or inoperable tumors and / or tumor metastases. Improved results have been observed when radiation therapy is combined with chemotherapy. Radiation therapy is based on the principle that delivering high doses of radiation to a target area will result in the death of reproductive cells in both tumor and normal tissues. Radiation administration regimens are usually defined in terms of radiation absorbed dose (Gy), time, and fractionation, and must be carefully prescribed by an oncologist. The amount of radiation a patient receives depends on various considerations, the two most important being the location of the tumor relative to other vital structures or organs in the body and the extent of tumor spread. A typical treatment course for patients undergoing radiation therapy is a 1-6 week treatment schedule, in which the patient receives a total dose ranging from 10-80 Gy in single daily doses of approximately 1.8-2.0 Gy, 5 days per week. In a preferred embodiment of the present invention, treating tumors in human patients with the combination therapy of the present invention and radiation is synergistic. In other words, tumor growth inhibition using substances comprising the combination of the present invention is enhanced when combined with radiation, optionally in combination with an additional chemotherapeutic or anticancer agent. Parameters for adjuvant radiotherapy are contained, for example, in WO 99 / 60023.

[0097] The type II anti-CD20 antibodies are administered to patients by known methods, intravenously as a bolus, by continuous infusion over an extended period of time, by intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intra-articular, intrasynovial, or intrathecal routes. Intravenous or subcutaneous administration of the antibodies is preferred.

[0098] The Bcl-2 inhibitor is administered to the patient in accordance with known methods, for example, intravenously as a bolus, by continuous infusion over an extended period of time, by intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, or oral routes. Intravenous, subcutaneous, or oral administration of the Bcl-2 inhibitor is preferred.

[0099] The present invention also relates to a kit comprising a type II anti-CD20 antibody and a selective Bcl-2 inhibitor for the combination therapy of patients suffering from CD20-expressing cancers.

[0100] In an embodiment of the invention, the kit further comprises a pharmaceutically acceptable carrier. The kit may further comprise a sterile diluent, preferably stored in a separate additional container. The kit may further comprise: It may include a package insert regarding the use of the combination therapy as a method for treating CD20-expressing cancer diseases, preferably B-cell non-Hodgkin's lymphoma (NHL).

[0101] The term "package insert" refers to the instructions typically found in the commercial packaging of a pharmaceutical product that may contain information about the indications, uses, dosage, administration, contraindications and / or warnings regarding the use of the pharmaceutical product.

[0102] In a preferred embodiment, the product container may further include a pharmaceutically acceptable carrier. The product may further include a sterile diluent, preferably stored in a separate additional container.

[0103] As used herein, "pharmaceutically acceptable carrier" is intended to include all materials compatible with pharmaceutical administration, such as solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and other materials and compounds compatible with pharmaceutical administration. Except insofar as any conventional media or substance is incompatible with the active compound, its use in the compositions of the present invention is contemplated. Supplementary active compounds may also be incorporated into the compositions.

[0104] Pharmaceutical Compositions and Methods Pharmaceutical compositions can be obtained by processing the type II anti-CD20 antibody or anti-Bcl-2 active substance according to the present invention with pharmaceutically acceptable inorganic or organic carriers. Examples of carriers that can be used for tablets, coated tablets, sugar-coated tablets, and hard gelatin capsules include lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc. Suitable carriers for soft gelatin capsules include vegetable oils, waxes, fats, semi-solid and liquid polyols, etc. However, for soft gelatin capsules, no carrier is usually required depending on the type of active substance. Suitable carriers for preparing solutions and syrups include water, polyols, glycerol, vegetable oils, etc. Suitable carriers for suppositories include natural or hardened oils, waxes, fats, semi-liquid or liquid polyols, etc.

[0105] The pharmaceutical composition may further contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavoring agents, salts for varying osmotic pressure, buffers, masking agents or antioxidants. The pharmaceutical composition may also contain other therapeutically valuable substances.

[0106] The pharmaceutical composition may further comprise one or more pharmaceutically acceptable carriers.

[0107] The present invention further provides a pharmaceutical composition, particularly for use in treating cancer, comprising (i) an effective first amount of a type II anti-CD20 antibody or (ii) an effective second amount of a selective Bcl-2 inhibitor, optionally including a pharmaceutically acceptable carrier and / or excipient.

[0108] Pharmaceutical compositions of type II anti-CD20 antibodies used alone according to the present invention are prepared by mixing the antibody having the desired purity for storage in the case of a lyophilized formulation or aqueous solution with any pharmaceutically acceptable carrier, excipient, or stabilizer (Remington's Pharmaceutical Sciences, 16th ed., Osol, A. Ed. (1980)). Acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the dosages and concentrations used, and include buffers such as phosphates, citrates, and other organic acids; antioxidants such as ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride, hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkylparabens, such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins These include proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates such as glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as Tween™, Pluronics™, or polyethylene glycol (PEG).

[0109] The pharmaceutical composition of an anti-Bcl-2 active substance alone, such as a Bcl-2 inhibitor, depends on its pharmaceutical properties. For example, in the case of small compounds such as ABT-737, ABT-199 or ABT-263, one formulation may be, for example:

[0110] a) Tablet formulation (wet granulation):

[0111] [Table 3]

[0112] Manufacturing Procedure: 1. Mix items 1, 2, 3 and 4 and granulate with purified water; 2. Dry the granules at 50℃; 3. Pass the granules through a suitable grinding device; 4. Add item 5, mix for 3 minutes and compress using a suitable press.

[0113] b) Capsule formulation:

[0114] [Table 4]

[0115] Manufacturing Procedure: 1. Mix items 1, 2 and 3 using a suitable mixer for 30 minutes; 2. Add items 4 and 5 and mix for 3 minutes; 3. Fill into suitable capsules.

[0116] In a further embodiment of the invention, the pharmaceutical composition according to the invention is two separate formulations directed to a type II anti-CD20 antibody and a Bcl-2 inhibitor.

[0117] The active ingredient may also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules), or in macroemulsions. The techniques are disclosed in Remington's Pharmaceutical Sciences, 16th Edition, Osol, A. Ed. (1980).

[0118] Sustained-release preparations may also be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles such as films or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactide (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and γ-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as Leupron Depot™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid.

[0119] The formulations to be used for in vivo administration must be sterile, which is readily accomplished by filtration through sterile filtration membranes.

[0120] In one embodiment, a method is provided in which an effective amount of GA-101 antibody is produced together with an effective amount of GDC-0199 to treat cancer.

[0121] In some embodiments, methods are provided in which an effective amount of GDC-0199 is prepared together with an effective amount of a GA-101 antibody to treat cancer.

[0122] In the provided production methods, the effective amount of the GA-101 antibody produced can be, for example, in the range of 500 mg to 1 g, or can be, for example, an effective amount discussed in the above section. In the provided production methods, the effective amount of GDC-0199 can be, for example, in the range of 20 mg to 1 g, or can be, for example, an effective amount discussed in the above section. The cancer to be treated can be, for example, a cancer discussed elsewhere herein.

[0123] The present invention relates, in part, to a method for treating a patient suffering from cancer, particularly a CD20-expressing cancer, in need of such treatment, comprising co-administering to said patient a type II anti-CD20 antibody and a selective Bcl-2 inhibitor, wherein the type II anti-CD20 antibody and the anti-Bcl-2 active agent are administered in effective amounts.

[0124] In one embodiment, the administration cycle is 28 days.

[0125] In certain embodiments of the methods of treating cancer in a patient provided herein, the method comprises administering to the patient a type II anti-CD20 antibody and a selective Bcl-2 antibody in one or more administration cycles. In one embodiment, each of the one or more administration cycles lasts at least one week. In another embodiment, each of the one or more administration cycles lasts at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks or more. In one embodiment, each administration cycle lasts 4 weeks.

[0126] In one embodiment, the therapeutic agent is administered to the patient during one administration cycle.

[0127] In another embodiment, the therapeutic agents are administered to the patient for two or more dosing cycles, e.g., 2, 3, 4, 5, 6, 7, or 8 or more dosing cycles. As an example, if the dosing cycle is 4 weeks and the patient is administered one or both therapeutic agents for 6 dosing cycles, the treatment regimen is 24 weeks, as illustrated in the dosing scheme shown in Figure 3.

[0128] In one embodiment, a method for treating cancer in a human in need thereof is provided, comprising administering to said patient multiple cycles of GA101 antibody and / or GDC-0199.

[0129] In one embodiment, both the GA101 antibody and GDC-199 are administered to the patient in one or more administration cycles over multiple administration cycles, and one of the GA101 antibody and GDC-0199 is administered in one or more administration cycles over multiple administration cycles.

[0130] In certain embodiments of the methods of treatment provided herein, the therapeutic agent is administered to a patient in a dosing scheme comprising two or three treatment phases, each treatment phase comprising at least one dosing cycle that is different from the dosing cycles of the other treatment phases. For example, in one embodiment in which the dosing cycles are four weeks, the type II anti-CD20 antibody may be administered to a patient once per week for two or more weeks in the first dosing cycle (e.g., the first treatment phase), and once per dosing cycle in the dosing cycles following the first dosing cycle (e.g., the second treatment phase).

[0131] In certain embodiments of the methods of treatment provided herein, the type II anti-CD20 antibody is administered to the patient once per week for at least one week of the administration cycle, and in some embodiments, when the administration cycle is two weeks or longer, the type II anti-CD20 antibody is administered to the patient once per administration cycle.

[0132] In certain embodiments of the methods of treatment provided herein, GDC-0199 is administered once daily during a dosing cycle.

[0133] Both GDC-0199 and the type II anti-CD20 antibody may be administered to the patient, for example, in dosing cycles, where only one of the therapeutic agents is administered to the patient in a dosing cycle.

[0134] In certain embodiments of the methods provided herein, in which a therapeutic agent is administered to a patient in a dosing scheme comprising multiple dosing cycles, the multiple dosing cycles comprise a first treatment phase having dosing cycles, in which a type II anti-CD20 antibody is administered once per dosing cycle and GDC-0199 is administered daily during the dosing cycle. Each dosing cycle in the first treatment phase can be, for example, four weeks. In some embodiments, the multiple dosing cycles can further comprise a second treatment phase (e.g., a maintenance phase) in which only a type II anti-CD20 antibody is administered to the patient or only GDC-0199 is administered to the patient.

[0135] In some embodiments of the methods provided herein, in which both a type II anti-CD20 antibody and GDC-0199 are administered to a patient during one or more administration cycles (e.g., in a treatment phase), the patient may subsequently be administered GDC-0199 alone (e.g., in a maintenance phase).

[0136] In certain embodiments in which only GDC-0199 is administered to a patient following combination therapy, GDC-0199 may be administered to a patient, for example, once daily, once every other day, once every 3, 4, 5, or 6 days, or once weekly.

[0137] In certain embodiments in which only a type II anti-CD20 antibody is administered to the patient after combination therapy, the type II anti-CD20 antibody may be administered to the patient, for example, once per week, once every two weeks, or once per month.

[0138] In certain embodiments of the methods of treatment provided herein, the amount of GDC-0199 administered per dose to a patient is increased during the first dosing cycle. For example, see Figure 2 for an exemplary dosing scheme in which the amount of GDC-0199 administered to a patient during the first dosing cycle is gradually increased from a 50 mg dose in week 1, to a 100 mg dose in week 2, to a 300 mg dose in week 3.

[0139] In some embodiments, the patient is administered escalating doses of GDC-0199 before the type II anti-CD20 antibody is administered, while in other embodiments, the patient is administered escalating doses of GDC-0199 after the type II anti-CD20 antibody is administered.

[0140] In some embodiments of the methods of treatment provided herein, the amount of GDC-0199 administered to a patient per dose is increased during the first dosing cycle from an initial dose ranging from 10 mg to 80 mg to a final dose ranging from 190 mg to 400 mg. In some embodiments, the initial amount of GDC-0199 administered to a patient per dose is 50 mg or 100 mg and is increased to 300 mg per dose. In some embodiments, the initial amount of GDC-0199 administered to a patient may be, for example, in the range of 20 mg to 60 mg (e.g., 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, or 60 mg doses), followed by 100 mg, 200 mg, 300 mg, or more of GDC-0199.

[0141] In certain embodiments of the methods provided herein, the patient is administered increasing doses of GDC-0199 prior to the first administration of a type II anti-CD20 antibody. In some embodiments, the patient is administered increasing doses of GDC-0199 after the first administration of a type II anti-CD20 antibody.

[0142] As used herein, the term "patient" typically refers to a human in need of treatment with a type II anti-CD20 antibody for any purpose (e.g., a patient suffering from a CD20-expressing cancer), and in one embodiment, to treat said cancer, or a precancerous condition or lesion. However, the term "patient" can also refer to non-human animals, preferably mammals, such as dogs, cats, horses, cows, pigs, sheep, and non-human primates, among others.

[0143] The present invention further includes type II anti-CD20 antibodies in combination with selective Bcl-2 inhibitors to treat CD20-expressing cancers.

[0144] The present invention further includes a type II anti-CD20 antibody in combination with a selective Bcl-2 inhibitor to treat patients suffering from CD20-expressing cancers.

[0145] The present invention further includes type II anti-CD20 antibodies and selective Bcl-2 inhibitors for use in the treatment of CD20-expressing cancers.

[0146] The present invention further includes a type II anti-CD20 antibody and a selective Bcl-2 inhibitor for use in treating patients suffering from CD20-expressing cancers.

[0147] In one embodiment, the selective Bcl-2 inhibitor is ABT-199.

[0148] In one embodiment, the type II anti-CD20 antibody has a ratio of binding ability of the type II anti-CD20 antibody to CD20 on Raji cells (ATCC-No. CCL-86) compared to rituximab of 0.3 to 0.6, in one embodiment 0.35 to 0.55, and in another embodiment 0.4 to 0.5.

[0149] In one embodiment, the type II anti-CD20 antibody is the GA101 antibody.

[0150] In one embodiment, the type II anti-CD20 antibody has high antibody-dependent cellular cytotoxicity (ADCC).

[0151] In some embodiments of the method for treating cancer in patients provided by the present invention, cancer is a non-solid tumor.In one embodiment, the non-solid tumor is a CD20-expressing non-solid tumor.Examples of non-solid tumors that can be treated by the method provided by the present invention include, for example, leukemia or lymphoma.In one embodiment, the non-solid tumor is a B-cell lymphoma.

[0152] In one embodiment, the CD20-expressing cancer is a B-cell non-Hodgkin's lymphoma (NHL).

[0153] In one embodiment, the type II anti-CD20 antibody is a monoclonal antibody.

[0154] The following examples, sequence listing and figures are provided to aid the understanding of the present invention, the true scope of which is set forth in the appended claims. It is understood that modifications can be made in the procedures set forth without departing from the spirit of the invention. [Example]

[0155] Example 1 Treatment of Lymphoma with a Combination of GDC-0199 and Obinutuzumab Preclinical data support the hypothesis that the combination of GDC-0199 and GA101 (in this case, obinutuzumab) exhibits greater antitumor activity than either drug administered alone. This study used a non-Hodgkin's lymphoma (NHL) xenograft model of aggressive lymphoma, the diffuse large B-cell lymphoma (DLBCL)-derived cell line SU.DHL-4. Obinutuzumab was administered intravenously at a dose of 1 mg / kg once weekly for 3 weeks, resulting in tumor stasis followed by delayed growth. GDC-0199 was administered at 100 mg / kg QD for 21 days, resulting in similar delayed growth. However, the combination of GDC-0199 and obinutuzumab induced a more-than-additive effect, resulting in tumor regression (5 of 8 partial regressions (PRs); see Figure 1). Three weeks of combination therapy resulted in a higher tumor growth inhibition (TGI) (118% TGI) compared to 76% (GA101) and 80% (GDC 0199) observed with single-agent administration (see Figure 1). When GA101 was combined with GDC 0199, higher tumor regressions (5PRs) were also observed compared to single-agent administration. In addition, TGI in the combination therapy group persisted after treatment was terminated on day 21, with 116% TGI observed at day 31 (10 days after the end of treatment) compared to 30% TGI for GA101 and 25% TGI for GDC 0199 as single agents. In summary, combining GA101 with GDC 0199 resulted in higher TGI and tumor regression compared to each agent administered separately in an NHL xenograft model.

[0156] In this study, the vehicles for GA-101 and GDC-0199 were saline and 60% phosal, respectively. Figure 1 displays the dose of GDC-0199 as the free base equivalent in mg / kg body weight. Results are displayed as approximate tumor volume versus time (days) as determined by linear mixed-effects modeling for each treatment group. Day 0 is the first day of treatment.

[0157] Example 2: Phase Ib multicenter study of GDC-0199 and obinutuzumab in patients with relapsed, refractory, or untreated chronic lymphocytic leukemia Two schedules will be evaluated: Schedule A (Figure 2), in which GDC-0199 is administered at escalating doses for 3 weeks before the first obinutuzumab infusion, and Schedule B (Figure 3), in which obinutuzumab is administered first followed by escalating dose levels of GDC-0199. Cohorts 1 for Schedule A and Schedule B will enroll in parallel. In addition, the dose-finding phase will evaluate whether administering GDC-0199 before the first obinutuzumab infusion (Schedule A) reduces the frequency of infusion reactions, thereby reducing the need for split doses of obinutuzumab and corticosteroid premedication.

[0158] The expansion phase will include two expansion cohorts of 20 patients each (relapsed / refractory and untreated CLL) to evaluate the safety and preliminary efficacy of selected combination doses and schedules.

[0159] In Schedule A, the combination therapy of GDC-0199 and obinutuzumab will be administered for a total of 7 cycles of 28 days each, including a total of 8 infusions of obinutuzumab and GDC-0199 QD.

[0160] In Schedule B, the combination of GDC-0199 and obinutuzumab is administered over a total of six cycles of 28 days each, for a total of nine infusions of obinutuzumab (eight doses; the first dose is split into two infusions) and GDC-0199 QD.

[0161] GDC-0199 monotherapy may be continued in patients beyond 6-7 cycles of the combination therapy described above (e.g., with acceptable toxicity and if a maximal clinical response (i.e., sustained improvement / reduction in tumor-bearing volume that has not yet stabilized for at least 2 months) has not yet been achieved). Patients may continue both GDC-0199 monotherapy and GDC-0199 monotherapy until a maximal response is achieved or up to 1 year after the last patient enrolled, whichever occurs first.

[0162] Example 3 Antitumor activity of combination therapy of GDC-0199 and type II anti-CD20 antibody (obinutuzumab) compared with combination therapy of GDC-0199 and type I anti-CD20 antibody (rituximab)

[0163] Test substance: The type II anti-CD20 antibody was GA101 antibody IgG1 (a chimeric humanized IgG1 antibody disclosed in WO 2005 / 044859 (referred to herein as B-HH6-B-KV1 GE, also known as obinutuzumab or RO5072759)) and was provided as a stock solution (concentration 9.4 mg / ml) by Roche GlycArt, Schlieren, Switzerland. The antibody buffer contained histidine, trehalose, and polysorbate 20. The antibody solution was appropriately diluted from stock with PBS before injection. GDC-0199 was obtained from Genentech Inc., California, USA.

[0164] Cell lines and culture conditions: The human Z138 mantle cell lymphoma cell line is routinely cultured in DMEM supplemented with 10% fetal bovine serum (PAA Laboratories, Austria) and 2 mM L-glutamine at 37°C in a water-saturated atmosphere with 5% CO. Cells were co-injected with MATRIGEL.

[0165] animal: Upon arrival (purchased from Charles River, Sulzfeld, Germany), 5-6 week-old female SCID beige mice were maintained under specific pathogen-free conditions with a 12-h light / dark daily cycle in accordance with the relevant guidelines (GV-Solas; Felasa; TierschG). The experimental study protocol was reviewed and approved by the local authorities (Regierung von Oberbayern; registration number 55.2-1-54-2531.2-26-09). After arrival, animals were maintained in an isolated section of the animal facility for one week for acclimatization and observation. Continuous health monitoring was performed at regular intervals. Food (Altromin Spezialfutter GmbH & Co., UK) and filtered water were provided ad libitum.

[0166] monitoring: Animals were monitored daily for clinical signs and adverse reactions. Animal body weights were recorded twice a week and tumor volumes were measured by calibus after staging for monitoring throughout the experiment.

[0167] Animal Treatment: Animal treatment began on the day of randomization, 18 days after tumor cell inoculation. RO5072759 or rituximab was administered as a single agent at a dose of 1 mg / kg ip once a week (days 18, 25, and 32) for 3 weeks. The corresponding vehicle was administered on the same days. GCD-0199 was administered at a dose of 100 mg / kg po once daily for 17 days (days 18 to 34). In the combination therapy group, antibody and GCD-0199 were administered at the same dose on the same day.

[0168] In vivo tumor growth inhibition studies: The effect of treatment on tumor volume expansion is shown in Figure 4. On day 35 after tumor cell inoculation, tumor growth inhibition of 32%, 59%, 73%, 96%, or 106% was observed in animals treated with rituximab, GDC-0199, RO5072759, the combination of GDC-0199 + rituximab, or the combination of GDC-0199 + RO5072759, respectively, compared to the control group.

[0169] Example 4: GDC-0199 administered as a single agent after combination with obinutuzumab significantly delays tumor regrowth This example describes results using the DLBCL SU-DHL-4 xenograft model discussed in Example 1 above. Initially, GDC-0199 was orally administered together with GA101 (in this example, obinutuzumab) at 1 mg / kg for 21 consecutive days for 3 weeks. The latter resulted in a high TGI (91%) compared to the 54% (GA101) and 24% (GDC-0199) TGI observed with each agent alone (Figure 5). On day 22, tumor-bearing mice in the combination cohort continued to receive GDC-0199 alone at 100 mg / kg for an additional 24 days. The latter resulted in a significant delay in tumor regrowth compared to mice treated with the combination of GA101 and GDC-0199 for 21 days (time to tumor progression in the combination cohort = 38 days vs. continued treatment with GDC-0199 = 45 days (Figure 5)). Thus, monotherapy with GDC-0199 in combination with GA101 has sustained efficacy in vivo. These results support the benefit of maintenance treatment with GDC-0199.

[0170] In Figure 5, controls are saline vehicle for GA101 and 60% saline vehicle for GDC-0199. GA101 was administered intravenously once weekly (QW) for 3 weeks, and GDC-0199 was administered orally daily (QD) for 21 days (QD x 21) either alone or in combination. As described above, a cohort of tumor-bearing mice received GDC-0199 alone for an additional 24 days after the combination treatment was terminated on day 21 (QD x 45). Below the x-axis, the treatment period is indicated by a solid black line for the combination cohort.

[0171] [ka] and continuous monotherapy with GDC-0199 is indicated by the black dashed line.

[0172] [ka] Shown in.

[0173] All publications and patent documents cited in this specification are incorporated by reference herein as if each individual publication or patent document was specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of example and illustration for purposes of clarity of understanding, it will be readily apparent to those skilled in the art that certain changes and modifications can be made in light of the teachings of the invention without departing from the spirit or scope of the appended claims.

Claims

1. and administering to a human in need thereof an effective amount of a GA101 antibody, or 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide, or a pharmaceutically acceptable salt thereof, for one or more dosing periods, followed by administration of an effective amount of the GA101 antibody or a pharmaceutically acceptable salt thereof. and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof during one or more administration periods.

2. A human in need of treatment is administered an effective amount of GA101 antibody, or 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide, or a pharmaceutically acceptable salt thereof, at 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 times the dose. for 14 days, followed by co-administration of effective amounts of the GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof during one or more administration periods.

3. The method of claim 1, comprising administering an effective amount of the GA101 antibody once per dosing period for 1, 2, 3, 4, 5, or 6 cycles, followed by co-administration of an effective amount of the GA101 antibody once per dosing period with 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof, one to three times daily during one or more dosing periods.

4. an effective amount of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof, administered 1 to 3 times daily for 1, 2, 3, 4, 5, or 6 dosing periods, followed by administration of an effective amount of the GA101 antibody.

2. The method of claim 1, comprising co-administering 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof to a subject once per administration period, from one to three times daily during one or more administration periods.

5. The method according to any one of claims 2 to 4, wherein the effective amount of the GA101 antibody is about 500 mg to about 3000 mg, and the effective amount of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof is about 20 mg to about 500 mg.

6. The effective doses of the GA101 antibody are 800, 900, 1000, 1100, 1200, 1300, 1400, and 1500 mg, and the effective doses of the GA101 antibody are 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4 5. The method of any one of claims 2 to 4, wherein the effective amount of the phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof is 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, or 300 mg.

7. The method of any one of claims 1 to 6, wherein the GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide or a pharmaceutically acceptable salt thereof are co-administered sequentially during each administration period, and each administration period is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days.

8. The method according to any one of claims 1 to 7, wherein the GA101 antibody is an anti-human CD20 antibody comprising HVR-H1 having the amino acid sequence of SEQ ID NO: 1, HVR-H2 having the amino acid sequence of SEQ ID NO: 2, HVR-H3 having the amino acid sequence of SEQ ID NO: 3, HVR-L1 having the amino acid sequence of SEQ ID NO: 4, HVR-L2 having the amino acid sequence of SEQ ID NO: 5, and HVR-L3 having the amino acid sequence of SEQ ID NO:

6.

9. The method of claim 8, wherein the GA101 antibody further comprises a VH domain comprising the amino acid sequence of SEQ ID NO:7 and a VL domain comprising the amino acid sequence of SEQ ID NO:

8.

10. The method of any one of claims 1 to 7, wherein the GA101 antibody comprises the amino acid sequence of SEQ ID NO:9 and the amino acid sequence of SEQ ID NO:

10.

11. The method of any one of claims 1 to 8, wherein the GA101 antibody is known as obinutuzumab.

12. The method of any one of claims 1 to 8, wherein the GA101 antibody comprises an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 9 and an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO:

10.

13. The method according to any one of claims 1 to 12, wherein the cancer is a CD20-expressing cancer.

14. The method of claim 13, wherein the cancer is a non-solid tumor.

15. 14. The method of claim 13, wherein the cancer is lymphoma or leukemia.

16. 14. The method of claim 13, wherein the leukemia is chronic lymphocytic leukemia (CLL).

17. 17. The method of claim 16, wherein the patient has relapsed, refractory, or untreated chronic lymphocytic leukemia.

18. and a method for treating cancer in a human in need thereof, comprising co-administering to said human an effective amount of a GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide during an administration period. the GA101 antibody is administered at 500 to 3000 mg weekly, and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide is administered at 50 to 300 mg once to three times daily during the administration period.

19. 1. A method of treating cancer in a human in need thereof, comprising administering to said human multiple cycles of GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide, wherein each cycle lasts at least 2, 3, 4, 5, or 6 weeks, and wherein the administration of 500 mg to 3000 mg of GA101 antibody is in a single dose. The method comprises administering the GA101 antibody once per administration cycle during one or more administration cycles of a plurality of administration cycles, and administering 10 mg to 300 mg of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide daily per administration cycle during one or more administration cycles of the plurality of administration cycles.

20. 20. The method of claim 19, wherein both the GA101 antibody and 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide are administered to the patient in multiple administration cycles of at least 2, 3, 4, 5, 6, 7, 8, or more than 8 administration cycles.

21. 20. The method of claim 19, wherein after a final administration cycle of the multiple administration cycles, a dose of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide is administered to the patient without administering a GA101 antibody to the patient.

22. 22. The method of claim 21, wherein the dose of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide administered to the patient in the absence of the GA101 antibody is from about 10 mg to about 300 mg of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide.

23. 23. The method of claim 22, wherein the dose of 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide administered to the patient in the absence of the GA101 antibody is administered to the patient for at least 3, 4, 5, 6, 7, 8 days, or 10 or more days, 20 or more days, or 30 or more days.

24. 20. The method of claim 19, wherein the multiple administration cycles comprise stepwise administration cycles wherein 2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)benzamide is administered to the patient in increasing daily doses during the stepwise administration cycles.

25. 25. The method of claim 24, wherein the escalating daily dose comprises an initial daily dose of 10 mg and a final daily dose of 300 mg.

26. The method according to any one of claims 19 to 25, wherein the cancer is a non-solid tumor.

27. The method of any one of claims 19 to 25, wherein the cancer is chronic lymphocytic leukemia (CLL).

28. 14. The method of claim 13, wherein the cancer is non-Hodgkin's lymphoma (NHL).

29. 14. The method of claim 13, wherein the cancer is acute myeloid leukemia (AML).

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