USO09539251B2
`
`(12) United States Patent
`Sampath et al.
`
`(10) Patent No.:
`(45) Date of Patent:
`
`US 9,539,251 B2
`Jan. 10, 2017
`
`(54) COMBINATION THERAPY OF A TYPE II
`ANT-CD2O ANTIBODY WITH A SELECTIVE
`BCL-2 INHIBITOR
`
`(71) Applicants: Genentech, Inc., South San Francisco,
`CA (US); F. HOFFMANN-LA
`ROCHE AG, Basel (CH)
`(72) Inventors: Deepak Sampath, South San Francisco,
`CA (US); Christian Klein, Iffeldorf
`(DE); Wayne John Fairbrother, South
`San Francisco, CA (US)
`(73) Assignees: Genentech, Inc., South San Francisco,
`CA (US); F. Hoffmann-La Roche AG,
`Basel (CH)
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 0 days.
`(21) Appl. No.: 14/020,761
`(22) Filed:
`Sep. 6, 2013
`
`(*) Notice:
`
`(65)
`
`Prior Publication Data
`US 2014/0248262 A1
`Sep. 4, 2014
`Related U.S. Application Data
`(60) Provisional application No. 61/698.379, filed on Sep.
`7, 2012.
`
`(2006.01)
`(2006.01)
`
`(51) Int. Cl.
`A 6LX 39/395
`A6 IK 3/496
`(52) U.S. Cl.
`CPC ................................... A6 IK3I/496 (2013.01)
`(58) Field of Classification Search
`USPC ....................................................... 424/133.1
`See application file for complete search history.
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`8/2010 Bruncko et al.
`7,767,684 B2
`5/2012 Bruncko et al.
`8,173,811 B2
`8,546,399 B2 10/2013 Bruncko et al.
`2005, 0123546 A1
`6, 2005 Umana et al.
`2008/0076779 A1
`3/2008 Elmore et al.
`2009/0098118 A1
`4/2009 Friess et al.
`2010/03051 22 A1* 12/2010 Bruncko et al. ........... 514,234.5
`2014/O11391.0 A1
`4/2014 Bruncko et al.
`
`FOREIGN PATENT DOCUMENTS
`
`WO
`WO
`
`2005044859 A2
`2010 138588 A2
`
`5, 2005
`12/2010
`
`OTHER PUBLICATIONS
`
`Herting et al. (Blood, 2010, 116: abstract # 3915, pp. 1-2).*
`Czuczman et al. (Leukemia & Lymphoma, Jun. 2010, 51(6): 983
`994).*
`Mossner et al. (Blood. 2010; 115(22): 4393-4402).*
`Taber's Cyclopedic Medical Dictionary (1985, F.A. Davis Com
`pany, Philadelphia, p. 274).*
`Busken, C et al. (Digestive Disease Week Abstracts and Itinerary
`Planner, 2003, abstract No: 850).*
`Kaiser (Science, 2006, 313: 1370).*
`International Searching Authority, “International Search Report for
`PCT Application No. PCT/US2013/058557, (Oct. 24, 2013), 4
`pageS.
`MacAuley, D., et al., “American Association for Cancer Research
`(AACR), 103rd, Annual meeting Chicago, Illinois,” (Mar. 31-Apr.
`4, 2012), Drugs of the Future 2012 Prous Science ESP (2012), vol.
`37, No. 6, pp. 451-455.
`Davids, M.S., et al., “Targeting the B-cell lymphoma leukemia 2
`family in cancer.” Journal of Clinical Oncology, (Sep. 1, 2012),
`American Society of Clinical Oncology, vol. 30, No. 25, pp.
`3127-3135.
`Sampath et al., “Abstract A245 Combination of the glycoengineered
`Type II CD20 antibody obinutuzumab (GA101), and the novel
`Bcl-2 selective inhibitor, ABT-199 (GDC-0199), results in superior
`in vitro and in vivo anti-tumor activity in models of B-cell malig
`nancies.” Mol Cancer Ther, 2013, 12 (11 Suppl).
`Sampath et al., “Combination of the glycoengineered Type II CD20
`antibody obinutuzumab (GA101) and The novel Bcl-2 selective
`Inhibitor GDC-0199 Results in Superior In Vitro and In Vivo
`Anti-tumor activity in models of B-Cell Malignancies,” Blood,
`2013,122 (21).
`Flinn et al., “Preliminary Results of a Phase 1b Study (GP28331)
`Combining GDC-0199 (ABT-199) and Obinutuzumab in Patients
`with Relapsed/Refractory or Previously Untreated Chronic
`Lymphocytic Leukemia,” Blood, 2014, 124(21).
`* cited by examiner
`Primary Examiner — Yan Xiao
`(74) Attorney, Agent, or Firm — Jones Day
`
`ABSTRACT
`(57)
`The present invention is directed to a combination therapy
`involving a type II anti-CD20 antibody and a selective Bcl-2
`inhibitor for the treatment of a patient suffering from cancer,
`particularly, a CD20-expressing cancer.
`
`14 Claims, 5 Drawing Sheets
`
`DRL EXHIBIT 1006 PAGE 1
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`DRL EXHIBIT 1006 PAGE 1
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`

`

`U.S. Patent
`
`Jan. 10, 2017
`
`Sheet 1 of 5
`
`US 9,539,251 B2
`
`FIG 1
`
`M- A) Wehicle (Saline + vehicle (60%hasai)
`re. - 8) GA (, tragikg
`mah C. GOC-(199, {}{rgikg
`wn - O GA 31, ingikg + GIC-898, (8-infkg
`
`3.
`
`
`
`St.
`
`2
`
`5.
`
`1000 i.
`
`SEC
`
`&
`
`DRL EXHIBIT 1006 PAGE 2
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`DRL EXHIBIT 1006 PAGE 2
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`

`

`U.S. Patent
`
`Jan. 10, 2017
`
`Sheet 2 of 5
`
`US 9,539,251 B2
`
`FIG 2
`
`Schedule A Dosing Schema: GDC-0199 Step-Up Dosing for
`Three Weeks Before Starting Obinutuzumab
`-H-0-0-0-HO-H-0- HO-HO-HO-H->
`
`GOC-O 99
`Dose
`Escalation
`
`O Obinutuzumab administration: W4 (C1D22), W5 (C2D1), W6 (C2D8), W9 (C3D1), W13 (C4D1),
`W17 (C5D1), W21(C6D1), and W25 (C7D1).
`A GDC-019950 mg dose (starting dose): W1 (C1D1 through C1D7)
`(O GDC-0199 100 mg dose: W2(C1D8 through C1D15)
`GDC-0199 300 mg dose (ending dose): W3 (C1D16) through W28 (C7D28)
`
`C = cycle, D= day; W = week.
`
`DRL EXHIBIT 1006 PAGE 3
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`DRL EXHIBIT 1006 PAGE 3
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`

`

`U.S. Patent
`
`Jan. 10, 2017
`
`Sheet 3 of 5
`
`US 9,539,251 B2
`
`FIG 3
`
`Schedule B Dosing Schema: Obinutuzumab Administered
`
`Before GDC-0199 Dose-Escalation
`
`Cohort 1
`
`C1
`wi
`
`C2
`C3
`C4
`C5
`cé
`W5
`w9
`W113
`W17
`wei
`
`
`GDC-0199 [
`Dose
`4
`Escalation | W2D2
`
`@ Obinutuzumab administration: W1 (C1, split dose on D1 and D2), W2 (C1D8), W3 (C1D15), z
`W5 (C2D1), W9 (C3D1), W13 (C4D1), W17 (C5D1), and W21 (C6D1)
`A GDC-0199 50 mg dose(starting dose): W2 (C1D9 through C1D14)
`© GDC-0199 100 mg dose: W3 (C1D15 through C1D21)
`
`CL GDC-0199 300 mg dose (ending dose): W4 (C1D22) through W24 (C6D28)
`
`Cohort 2
`
`
`4¢o¢/¢1-¢41++-¢++++e_ +H .
`C1
`C2
`C3
`C4
`c5
`cé6
`
`W5
`wg
`W13
`W17
`W21
`
`
`_w4
`GDC-0199/
`Dose
`4
`Escalation | W2D2
`
`@ Obinutuzumab administration: W1 (C1, split dose on D1 and D2), W2 (C1D8), W3 (C1D15),
`W5(C2D1), W9 (C3D1), W13 (C4D1), W17 (C5D1), and W21 (C6D1)
`© GDC-0199 100 mg dose (starting dose): W2 (C1D9 through C1D14)
`
`L] GDC-0199 300 mg dose (ending dose): W3 (C1D15) through W24 (C6D28)
`
`Cohort3
`
`
`ooejele+++¢e+-+++ ¢e+++¢eH >
`Ci
`C2
`C3
`c4
`C5
`C6
`W5
`wg
`W13
`W17
`W21
`
`
`Ol
`
`W2D2
`
`GDC-0199
`Dose
`Escalation
`
`@ Obinutuzumab administration: W1 (C1, split dose on D1 and D2), W2 (C1D8), W3 (C1D15), W5 (C2D1),
`W9 (C3D1), W13 (C4D1), W17 (C5D1), and W21 (C6D1)
`LJ GDC-0199 300 mg dose: W2 (C1D9) through W24 (C6D28)
`
`C=cycle; D=day; W=week.
`Schedule B includes approximately three dosing cohorts to be enrolled sequentially.
`
`DRL EXHIBIT 1006 PAGE 4
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`DRL EXHIBIT 1006 PAGE 4
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`DRL EXHIBIT 1006 PAGE 4
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`

`

`U.S. Patent
`
`Jan. 10, 2017
`
`Sheet 4 of 5
`
`US 9,539,251 B2
`
`FIG 4
`
`
`i) —— Vehicle ip.
`
`4000 . 805072759; Imaike ip; Day 18, 25, 22
`
`
`0 + —O— Aituximab; lnig/kg Le. Day 28 25, 22
`
`3000 -) —a—600-0199; 100me/kg p.9.; Day 18 - 24
`
`
`Rituximab: imge/ke ip. Day 18, 25, 32
`
`QO
`
`4 —M@ GDC-0199; 100mg/kg p.9.: Day 18 - 34
`:
`ROSOF2759; ima/kg ips Day 18, 25, 32
`
`Ch) me GEC; LOOMgH 9.9.; Day 1B 34 Lge gated ecssenneeseeaglce ce
`:
`
`
`TVimm*)
`
`DRL EXHIBIT 1006 PAGE 5
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`DRL EXHIBIT 1006 PAGE 5
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`DRL EXHIBIT 1006 PAGE 5
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`

`

`U.S. Patent
`
`Jan. 10, 2017
`
`Sheet S of 5
`
`US 9,539,251 B2
`
`FGS
`
`4000
`
`3 O O O
`
`2000
`
`-e- Control
`wA GA-1011 mg/kg, QWX3
`-0- GDC-0199 100mg/kg, QDx21
`A GA101 - GDC-0199 ODX21
`ahe GA101 + GDC-0199 CDX45
`s
`
`2.
`
`1000-
`
`
`
`21 1 /
`
`A
`
`GA-101 + GDC-0199 me GDC-0199 only
`
`DRL EXHIBIT 1006 PAGE 6
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`DRL EXHIBIT 1006 PAGE 6
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`

`

`1.
`COMBINATION THERAPY OF A TYPE II
`ANT-CD2O ANTIBODY WITH A SELECTIVE
`BCL-2 INHIBITOR
`
`This application claims priority to U.S. Provisional No.
`61/698.379, filed Sep. 7, 2012, the content of which is
`incorporated herein by reference in its entirety.
`
`TECHNICAL FIELD
`
`The present invention is directed to a combination therapy
`involving a type II anti-CD20 antibody and a selective Bcl-2
`inhibitor for the treatment of a patient suffering from cancer,
`particularly a CD20-expressing cancer.
`
`5
`
`10
`
`15
`
`BACKGROUND
`
`25
`
`30
`
`35
`
`45
`
`The CD20 molecule (also called human B-lymphocyte
`restricted differentiation antigen or Bp35) is a hydrophobic
`transmembrane protein with a molecular weight of approxi
`mately 35 kD located on pre-B and mature B lymphocytes
`(Valentine, M. A., et al., J. Biol. Chem. 264 (19) (1989)
`11282-11287; and Einfield, D. A., et al. EMBO J. 7(3)
`(1988) 711-717). CD20 is found on the surface of greater
`than 90% of B cells from peripheral blood or lymphoid
`organs and is expressed during early pre-B cell development
`and remains until plasma cell differentiation. CD20 is pres
`ent on both normal B cells as well as malignant B cells. In
`particular, CD20 is expressed on greater than 90% of B cell
`non-Hodgkin’s lymphomas (NHL) (Anderson, K. C., et al.,
`Blood 63(6) (1984) 1424-1433) but is not found on
`hematopoietic stem cells, pro-B cells, normal plasma cells,
`or other normal tissues (Tedder, T. F., et al., J. Immunol.
`135(2) (1985) 973-979).
`The 85 amino acid carboxyl-terminal region of the CD20
`protein is located within the cytoplasm. The length of this
`40
`region contrasts with that of other B cell-specific surface
`structures Such as IgM, Ig), and IgG heavy chains or
`histocompatibility antigens class I1 a or B chains, which
`have relatively short intracytoplasmic regions of 3, 3, 28, 15,
`and 16 amino acids, respectively (Komaromy, M., et al.
`NAR 11 (1983) 6775-6785). Of the last 61 carboxyl-termi
`nal amino acids. 21 are acidic residues, whereas only 2 are
`basic, indicating that this region has a strong net negative
`charge. The GenBank Accession No. is NP-690605. It is
`thought that CD20 might be involved in regulating an early
`step(s) in the activation and differentiation process of B cells
`(Tedder, T. F., et al., Eur. J. Immunol. 16 (1986) 881-887)
`and could function as a calcium ion channel (Tedder. T. F.,
`et al., J. Cell. Biochem. 14D (1990) 195).
`There exist two different types of anti-CD20 antibodies
`which differ significantly in their mode of CD20 binding and
`biological activities (Cragg, M. S., et al., Blood 103 (2004)
`2738-2743; and Cragg, M. S., et al., Blood 101 (2003)
`1045-1052). Type I antibodies, as e.g. rituximab, are potent
`in complement mediated cytotoxicity, whereas type II anti
`bodies, as e.g. Tositumomab (B1), 11B8, AT80 or human
`ized B-Ly1 antibodies, effectively initiate target cell death
`via caspase-independent apoptosis with concomitant phos
`phatidylserine exposure.
`The shared common features of type I and type II anti
`CD20 antibodies are summarized in Table 1 below.
`
`50
`
`55
`
`60
`
`65
`
`US 9,539,251 B2
`
`2
`TABLE 1.
`
`Properties of type I and type II anti-CD20 antibodies
`
`type I anti-CD20 antibodies
`type I CD20 epitope
`Localize CD20 to lipid rafts
`Increased CDC (if IgG1 isotype)
`ADCC activity (if IgG1 isotype)
`Full binding capacity
`Homotypic aggregation
`Apoptosis induction upon cross
`linking
`
`type II anti-CD20 antibodies
`type II CD20 epitope
`Do not localize CD20 to lipid rafts
`Decreased CDC (if IgG1 isotype)
`ADCC activity (if IgG1 isotype)
`Reduced binding capacity
`Stronger homotypic aggregation
`Strong cell death induction without
`cross-linking
`
`The Bcl-2 family of proteins regulates programmed cell
`death triggered by developmental cues and in response to
`multiple Stress signals (Cory. S., and Adams, J. M., Nature
`Reviews Cancer 2 (2002) 647-656: Adams, Genes und
`Development 17 (2003). 2481-2495; Danial, N. N., and
`Korsmeyer, S.J., Cell 116 (2004) 205-219). Whereas cell
`survival is promoted by Bcl-2 itself and several close
`relatives (Bcl-XL, Bcl-W. Mcl-1 and A1), which bear three or
`four conserved Bcl-2 homology (BH) regions, apoptosis is
`driven by two other sub-families. The initial signal for cell
`death is conveyed by the diverse group of BH3-only pro
`teins, including Bad, Bid, Bim, Puma and Noxa, which have
`in common only the Small BH3 interaction domain (Huang
`and Strasser, Cell 103 (2000) 839-842). However, Bax or
`Bak, multi-domain proteins containing BH1-BH3, are
`required for commitment to cell death (Cheng, et al.,
`Molecular Cell 8 (2001) 705-711; Wei, M. C., et al., Science
`292 (2001) 727-730; Zong, W. X., et al., Genes and Devel
`opment 15 148 (2001) 1-1486). When activated, they can
`permeabilize the outer membrane of mitochondria and
`release pro-apoptogenic factors (e.g. cytochrome C) needed
`to activate the caspases that dismantle the cell (Wang, K.,
`Genes and Development 15 (2001) 2922-2933; (Adams,
`2003 Supra); Green, D. R., and Kroemer, G., Science 305
`(2004) 626-629).
`Interactions between members of these three factions of
`the Bcl-2 family dictate whether a cell lives or dies. When
`BH3-only proteins have been activated, for example, in
`response to DNA damage, they can bind via their BH3
`domain to a groove on their pro-survival relatives (Sattler, et
`al., Science 275 (1997) 983-986). How the BH3-only and
`Bcl-2-like proteins control the activation of Bax and Bak,
`however, remains poorly understood (Adams, 2003 supra).
`Most attention has focused on Bax. This soluble monomeric
`protein (Hsu, Y. T., et al., Journal of Biological Chemistry
`272 (1997) 13289-1 3834; Wolter, K. G., et al., Journal of
`Cell Biology 139 (1997) 1281-92) normally has its mem
`brane targeting domain inserted into its groove, probably
`accounting 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 Bio 1 164
`(2004) 1021-1032). Several unrelated peptides/proteins
`have been proposed to modulate Bax activity, reviewed in
`Lucken-Ardjomande, S., and Martinou, J. C. J. Cell Sci 118
`(2005) 473–483, but their physiological relevance remains to
`be established. Alternatively, Bax may be activated via
`direct engagement by certain BH3-only proteins (Lucken
`Ardjomande, S., and Martinou, J. C. 2005 supra), the best
`documented being a truncated form of Bid, tBid (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 discussed elsewhere
`(Adams 2003 supra), the oldest model, in which Bcl-2
`
`DRL EXHIBIT 1006 PAGE 7
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`DRL EXHIBIT 1006 PAGE 7
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`US 9,539,251 B2
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`3
`directly engages Bax (Oltvai, Z. N., et al., Cell 74 (1993)
`609-619), has become problematic because Bcl-2 is mem
`brane bound while Bax is cytosolic, and their interaction
`seems highly dependent on the detergents used for cell lysis
`(Hsu, Y.T., and Youle, 1997 supra). Nevertheless, it is well
`established that the BH3 region of Bax can mediate asso
`ciation with Bcl-2 (Zha, H. and Reed, J., Journal of Bio
`logical Chemistry 272 (1997) 31482-88: Wang, K., et al.,
`Molecular und Cellular Biology 18 (1998) 6083-6089) and
`that Bcl-2 prevents the oligomerization of Bax, even though
`no heterodimers can be detected (Mikhailov, V., et al.,
`Journal of Biological Chemistry 276 (2001) 18361-18374).
`Thus, whether the pro-survival proteins restrain Bax activa
`tion directly or indirectly remains uncertain.
`Although Bax and Bak seem in most circumstances to be
`functionally equivalent (Lindsten, T. et al., Molecular Cell 6
`(2000) 1389-1399; Wei, M. C., et al., 2001 supra), substan
`tial differences in their regulation would be expected from
`their distinct localization in healthy cells. Unlike Bax, which
`is largely cytosolic, Bak resides in complexes on the outer
`membrane of mitochondria and on the endoplasmic reticu
`lum of healthy cells (Wei, M. C., et al., 2000 supra; Zong,
`W. X., et al., Journal of Cell Biology 162 (2003) 59-69).
`Nevertheless, on receipt of cytotoxic signals, both Bax and
`Bak change conformation, and Bax translocates to the
`organellar membranes, where both Bax and Bak then form
`homo-oligomers that can associate, leading to membrane
`permeabilization (Hsu. Y.T. et al., PNAS 94 (1997) 3668
`3672; Wolter, K. G. et al., 1997 Supra; 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, M. C., et al., 2001 supra; Mikhailov, V., et
`al., Journal of Biological Chemistry 278 (2003).5367-5376).
`There exist various Bcl-2 inhibitors, which all have the
`same property of inhibiting prosurvival members of the
`Bcl-2 family of proteins and are therefore promising candi
`dates for the treatment of cancer. Such Bcl-2 inhibitors are
`e.g. 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,
`40
`2-Methoxyantimycin A, HA-14-1. KF-67544, Purpurogal
`lin, TP-TW-37, YC-137 and Z-24, and are described e.g. in
`Zhai, D., et al., Cell Death and Differentiation 13 (2006)
`1419-1421.
`Smith, M. R., et al. Molecular Cancer Therapeutics 3 (12)
`(2004) 1693-1699 and Ramanarayanan, J. et al., British
`Journal of 1-laematology 127(5) (2004) 519-530, refer to a
`combination of a type I anti-CD20 antibody (rituximab) with
`antisense Bcl-2 oligonucleotides (Oblimersen).
`
`10
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`15
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`25
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`30
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`35
`
`4
`In an embodiment, the cancer is a non-Solid tumor.
`In certain embodiments, methods are provided for the
`treatment of a cancer in a human in need thereof comprising
`administering to said human a GA101 antibody and/or
`GDC-0199 in multiple dosing cycles. In an embodiment,
`each dosing cycle of the multiple dosing cycle is for at least
`1 week. In an embodiment, each dosing cycle of the multiple
`dosing cycle is for at 2, for at least 3, for at least 4, for at least
`5, or for at least 6 weeks.
`In an embodiment wherein the GA101 antibody and
`GDC-0199 are administered to the human in multiple dosing
`cycles, GA101 antibody can, for example, be administered
`once per dosing cycle for one or more dosing cycles of the
`multiple dosing cycles. The amount of GA101 administered
`per dose can, for example, be between about 300 mg to
`about 3000 mg, or between about 500 mg to about 3000 mg.
`or about 500 mg to about 1200 mg.
`In an embodiment wherein the GA101 antibody and
`GDC-0199 are administered to the human in multiple dosing
`cycles, GDC-0199 can, for example, be administered each
`day in a dosing cycle for one or more dosing cycles of the
`multiple dosing cycles. In an embodiment, GDC-0199 is
`administered in fewer than all of the days of the initial
`dosing cycle, and is administered each day of the dosing
`cycles of the multiple dosing cycles that follow the initial
`dosing cycle. The amount of GDC-0199 administered per
`day can be between about 10 mg to about 1,000 mg, about
`20 mg to about 800 mg, about 20 mg to about 500 mg. or
`between about 50 mg to about 300 mg.
`In an embodiment, both the GA101 antibody and GDC
`0199 are administered to the patient in at least 2, 3, 4, 5, 6,
`7, 8, or more than 8, dosing cycles of the multiple dosing
`cycles.
`In certain embodiments of the methods provided for the
`treatment of a cancer in a human in need thereof comprising
`administering to said human both a GA101 antibody and
`GDC-0199 in multiple dosing cycles, following the last
`dosing cycle of multiple dosing cycles, GDC-0199 alone can
`be administered to the human in the absence of the GA101
`antibody, or the GA101 antibody alone can be administered
`to the patient in the absence of GDC-0199. For instance,
`when GDC-0199 is administered alone to the human (e.g.,
`following the last cycle of multiple dosing cycles wherein
`both GDC-0199 and the GA101 antibody are administered
`to the human), GDC-0199 can be administered to the human
`for at least 3, 4, 5, 6, 7, 8 or 9 days, or for 10 or more days,
`for 20 or more days, or for 30 or more days.
`In yet another embodiment of the methods provided
`wherein a GA101 antibody and GDC-0199 are administered
`to the patient in multiple dosing cycles, the multiple dosing
`cycles comprise an escalating dosing cycle in which GDC
`0199 is administered to the patient in escalating daily dose
`amounts during the escalating dosing cycle.
`
`DESCRIPTION OF THE FIGURES
`
`FIG. 1. Antitumor activity of combined treatment of a
`type II anti-CD20 antibody (GA101 antibody, in this case,
`obinutuzumab) with a Bcl-2 inhibitor (ABT-199, a.k.a.
`GDC-0199). Arrows and line under the x-axis indicates the
`days of dosing of GA101 and GDC-0199, respectively.
`FIG. 2. Exemplary dosing schedule for administering
`GDC-199 with obinutuZumab.
`FIG. 3. Exemplary dosing schedule for administering
`GDC-199 with obinutuZumab.
`FIG. 4. Antitumor activities of a type II anti-CD20
`antibody (obinutuzumab, a.k.a. R05072759) used alone or in
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`SUMMARY OF THE INVENTION
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`Provided herein are methods for the treatment of a patient
`Suffering from cancer, comprising co-administering, to a
`patient in need of such treatment, a type II anti-CD20
`antibody and a selective Bcl-2 inhibitor. The co-administra
`tion may be simultaneous or sequential in either order.
`An example of the type II anti-CD20 antibody for use in
`the present invention is a GA101 antibody.
`In an embodiment, the type II anti-CD20 antibody has
`increased antibody dependent cellular cytotoxicity (ADCC).
`In an embodiment, at least 40% of the oligosaccharides of
`the Fc region of said type II anti-CD20 antibody are non
`fucosylated.
`In an embodiment, the selective Bcl-2 inhibitor is GDC
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`0199 (also known as ABT-199), or a pharmaceutically
`acceptable salt thereof.
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`combination with GDC-0199, and of a type I anti-CD20
`antibody (rituximab) used alone or in combination with
`GDC-0199 on human Z138 mantle cell lymphoma cells.
`FIG. 5. Results from xenograft model of aggressive
`lymphoma demonstrating that single agent treatment with
`GDC-0199 following combination of GDC-0199 with type
`II anti-CD20 antibody (GA101 antibody, in this case, obi
`nutuZumab) delays tumor regrowth.
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`
`The present invention relates to the method described
`above.
`The present invention also relates to a method for the
`treatment of a human in need thereof comprising adminis
`tering to said human an effective amount of a
`GA101 antibody or 2-(1H-pyrrolo2,3-bipyridin-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
`O
`a
`pharmaceutically acceptable salt thereof for one or more
`dosing periods, followed by co-administering an effective
`amount of said GA101 antibody and 2-(1H-pyrrolo2,3-b
`pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethyl
`cyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tet
`rahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)
`benzamide or a pharmaceutically acceptable salt thereof for
`one or more dosing periods.
`The present invention also relates to a method for the
`treatment of a human in need thereof comprising adminis
`tering to said human an effective amount of a GA101
`antibody or 2-(1H-pyrrolo2,3-bipyridin-5-yloxy)-4-(4-((2-
`(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl)methyl)pip
`erazin-1-yl)-N-(3-nitro-4-((tetrahydro-2H-pyran-4-yl)meth
`ylamino)phenylsulfonyl)benzamide for 0, 1, 2, 3, 4, 5, 6, 7,
`8, 9, 10, 11, 12, 13, or 14 days, followed by co-administering
`an effective amount of said GA101 antibody and 2-(1H
`pyrrolo2,3-bipyridin-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.
`The present invention also relates to a method for the
`treatment of a human in need thereof comprising adminis
`tering an effective amount of said GA101 antibody for 1, 2,
`3, 4, 5, 6, or 7 days, followed by co-administering an
`effective amount of said GA101 antibody antibody and
`2-(1H-pyrrolo2,3-bipyridin-5-yloxy)-4-(4-((2-(4-chloro
`phenyl)-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.
`The present invention also relates to a method for the
`treatment of a human in need thereof comprising adminis
`tering an effective amount of 2-(1H-pyrrolo2,3-bipyridin
`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 co-administering an effective
`amount of said GA101 antibody g and 2-(1H-pyrrolo2,3-
`hpyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethyl
`cyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tet
`rahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)
`benzamide or a pharmaceutically acceptable salt thereof for
`one or more dosing periods.
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`The present invention also relates to a method for the
`treatment of a human in need thereof comprising adminis
`tering an effective amount of said GA101 antibody once
`every dosing period for 1, 2, 3, 4, 5 or 6 cycles, followed by
`co-administering
`an
`effective
`amount
`of said
`GA101 antibody antibody once every dosing period and
`2-(1H-pyrrolo2,3-bipyridin-5-yloxy)-4-(4-((2-(4-chloro
`phenyl)-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, two or three times a day for one or more
`dosing periods.
`The present invention also relates to a method for the
`treatment of a human in need thereof comprising adminis
`tering an effective amount of 2-(1H-pyrrolo2,3-bipyridin
`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, two or three
`times a day for 1, 2, 3, 4, 5 or 6 dosing periods, followed by
`co-administering an effective amount of said GA101 anti
`body once every dosing period and 2-(1H-pyrrolo2,3-b
`pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethyl
`cyclohex-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, two or three times a day for one or more dosing periods.
`The present invention also relates to any one of the above
`methods, wherein the effective amount of said
`GA101 antibody 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 and the effective amount of 2-(1H-pyrrolo2,3-b
`pyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethyl
`cyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-((tet
`rahydro-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, 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, or 1000 mg.
`The present invention also relates to any one of the above
`methods, wherein the effective amount of said
`GA101 antibody is 800, 900, 1000, 1100, 1200, 1300, 1400
`or 1500 mg, and the effective amount of 2-(1H-pyrrolo2,
`3-bipyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimeth
`ylcyclohex-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
`or 300 mg.
`The present invention also relates to any one of the above
`methods, wherein when said cancer is NHL, the effective
`amount of said GA101 antibody is 800, 900, 1000, 1100.
`1200, 1300, 1400 or 1500 mg, and the effective amount of
`2-(1H-pyrrolo2,3-bipyridin-5-yloxy)-4-(4-((2-(4-chloro
`phenyl)-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,
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`7
`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, and 800 mg.
`The present invention also relates to any one of the above
`methods, wherein when said cancer is AML, the effective
`amount of said GA101 antibody is 800, 900, 1000, 1100,
`1200, 1300, 1400 or 1500 mg, and the effective amount of
`2-(1H-pyrrolo2,3-bipyridin-5-yloxy)-4-(4-((2-(4-chloro
`phenyl)-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, 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, and 800 mg.
`The present invention also relates to any one of the above
`methods, wherein said GA101 antibody and 2-(1H-pyrrolo
`2,3-bipyridin-5-yloxy)-4-(4-((2-(4-chlorophenyl)-4,4-dim
`ethylcyclohex-1-enyl)methyl)piperazin-1-yl)-N-(3-nitro-4-
`((tetrahydro-2H-pyran-4-yl)methylamino)phenylsulfonyl)
`benzamide or a pharmaceutically acceptable salt thereof
`were co-administered sequentially during each dosing
`period, and each dosing period is 5, 6, 7, 8, 9, 10, 11, 12, 13,
`or 14 days.
`The term “antibody' herein is used in the broadest sense
`and encompasses various antibody structures, including but
`not limited to monoclonal antibodies, polyclonal antibodies,
`multispecific antibodies (e.g., bispecific antibodies), and
`antibody fragments so long as they exhibit the desired
`antigen-binding activity.
`The term “monoclonal antibody” as used herein refers to
`an antibody obtained from a population of Substantially
`homogeneous antibodies, i.e., the individual antibodies
`comprising the population are identical and/or bind the same
`epitope, except for possible variant antibodies, e.g., contain
`ing naturally occurring mutations or arising during produc
`tion of a monoclonal antibody preparation, such variants
`generally being present in minor amounts. In contrast to
`polyclonal antibody preparations, which typically include
`different antibodies directed against different determinants
`(epitopes), each monoclonal antibody of a monoclonal anti
`body preparation is directed against a single determinant on
`an antigen. Thus, the modifier "monoclonal indicates the
`character of the antibody as being obtained from a Substan
`tially homogeneous population of antibodies, and is not to be
`construed as requiring production of the antibody by any
`particular method. For example, the monoclonal antibodies
`to be used in accordance with the present invention may be
`made by a variety of techniques, including but not limited to
`the hybridoma method, recombinant DNA methods, phage
`display methods, and methods utilizing transgenic animals
`containing all or part of the human immunoglobulin loci,
`Such methods and other exemplary methods for making
`monoclonal antibodies being described herein.
`In one embodiment, said type II anti-CD20 antibody is a
`monoclonal antibody.
`The term "chimeric antibody” refers to a monoclonal
`antibody comprising a variable region, i.e., binding region,
`from one source or species and at least a portion of a
`constant region derived from a different source or species,
`usually prepared by recombinant DNA techniques. Chimeric
`antibodies comprising a murine variable region and a human
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`constant region are especially preferred. Such murine/hu
`man chimeric antibodies are the product of expressed immu
`noglobulin genes comprising DNA segments encoding
`murine immunoglobulin variable regions and DNA seg
`ments encoding human immunoglobulin cons