Europalsches
`Patentamt
`European
`Patent Office
`Office europeen
`des brevets
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`(19)
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`(12)
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`111111111111111111111111111111111111111111111111111111111111111111111111111
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`EP 1 210 115 81
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`EUROPEAN PATENT SPECIFICATION
`
`(45) Date of publication and mention
`of the grant of the patent:
`05.08.2009 Bulletin 2009/32
`
`(21) Application number: 00959423.5
`
`(22) Date of filing: 25.08.2000
`
`(51) lnt Cl.:
`A61K 39/395(2006·01)
`
`C07K 16/32(2006·01)
`
`(86) International application number:
`PCT/US2000/023391
`
`(87) International publication number:
`WO 2001/015730 (08.03.2001 Gazette 2001/10)
`
`(54) DOSAGES FOR TREATMENT WITH ANTI-ErbB2 ANTIBODIES
`
`DOSIERUNG FUR DIE BEHANDLUNG MIT ANTI ERBB2-ANTIKQRPERN
`
`DOSAGES POUR TRAITEMENT AVEC DES ANTICORPS ANTI-ErbB2
`
`(84) Designated Contracting States:
`AT BE CH CY DE DK ES Fl FR GB GR IE IT Ll LU
`MC NL PT SE
`Designated Extension States:
`AL LT LV MK RO Sl
`
`(30) Priority: 27.08.1999 US 151018 P
`23.06.2000 us 213822 p
`
`(43) Date of publication of application:
`05.06.2002 Bulletin 2002/23
`
`(73) Proprietor: Genentech, Inc.
`South San Francisco CA 94080-4990 (US)
`
`(72) Inventors:
`• BAUGHMAN, Sharon, Ann
`Ventura, CA 93001 (US)
`• SHAK, Steven
`Burlingame, CA 94010 (US)
`
`(74) Representative: Walton, Sean Malcolm et al
`Mewburn Ellis LLP
`33 Gutter Lane
`London
`EC2V BAS (GB)
`
`(56) References cited:
`WO-A-98/17797
`WO-A2-01/00245
`US-A- 5 055 393
`
`WO-A-99/31140
`WO-A2-01/00245
`
`• PEGRAM MARK D ET AL: "Phase II study of
`receptor-enhanced chemosensitivity using
`recombinant humanized anti-p185HER2/neu
`monoclonal antibody plus cisplatin in patients
`with H ER2/neu-overexpressi ng metastatic breast
`cancer refractory to chemotherapy treatment."
`JOURNAL OF CLINICAL ONCOLOGY, vol. 16, no.
`8, August 1998 (1998-08), pages 2659-2671,
`XP000971446 ISSN: 0732-183X
`• GOLDENBERG M M: "Trastuzumab, a
`recombinant DNA-derived humanized
`monoclonal antibody, a novel agent for the
`treatment of metastatic breast cancer" CLINICAL
`THERAPEUTICS,EXCERPTA MEDICA,
`PRINCETON, NJ,US, vol. 21, no. 2, 1999, pages
`309-318, XP000918210 ISSN: 0149-2918
`• PEDLEY R.B.: 'Pharmacokinetics of monoclonal
`antibodies. Implications for their use in cancer
`therapy' CLINICAL IMMUNOTHERAPEUTICS vol.
`6, no. 1, 01 January 1996, ADIS INTERNATIONAL,
`AUCKLAND, NZ, pages 54-67, XP009100791
`• REILLY R.M. ET AL: 'Problems of delivery of
`monoclonal antibodies. Pharmaceutical and
`pharmacokinetic solutions' CLINICAL
`PHARMACOKINETICS vol. 28, no. 2, 01 January
`1995, ADIS INTERNATIONAL LTD., AUCKLAND,
`NZ, pages 126 - 142, XP0091 00787
`• SHARIFI J. ET AL: 'Improving monoclonal
`antibody pharmacokinetics via chemical
`modification' QUARTERLY JOURNAL OF
`NUCLEAR MEDICINE vol. 42, no. 4, 01 January
`1998, MILAN, IT, pages 242-249, XP009100792
`
`Remarks:
`The file contains technical information submitted after
`the application was filed and not included in this
`specification
`
`Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent
`Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the
`Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been
`paid. (Art. 99(1) European Patent Convention).
`
`c.
`w
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`Description
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`Field of the Invention
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`[0001] The present disclosure concerns the treatment of disorders characterized by the overexpression of ErbB2 or
`disorders expressing epidermal growth factor receptor (EGFR), comprising administering to a human or animal presenting
`the disorders a therapeutically effective amount of an antibody that binds ErbB2. More specifically, the disclosure concerns
`the treatment of human patients susceptible to or diagnosed with cancer overexpressing ErbB2 or expressing EGFR,
`where the treatment is with an anti-ErbB2 antibody administered by front loading the dose of antibody during treatment
`by intravenous and/or subcutaneous administration. The disclosure optionally includes treatment of cancer in a human
`patient with a combination of an anti-ErbB2 antibody and a chemotherapeutic agent, such as, but not limited to, a taxoid.
`The taxoid may be, but is not limited to paclitaxel or docetaxel. The disclosure further includes treatment of cancer in a
`human patient with a combination of anti-ErbB2 antibody and a chemotherapeutic agent, such as, but not limited to, an
`anthracycline derivative. Optionally, treatment with a combination of anti-ErbB2 and an anthracycline derivative includes
`treatment with an effective amount of a cardioprotectant. The present invention further concerns infrequent dosing of
`anti-ErbB2 antibodies.
`
`Background of the Invention
`
`[0002] Proto-oncogenes that encode growth factors and growth factor receptors have been identified to play important
`roles in the pathogenesis of various human malignancies, including breast cancer. It has been found that the human
`ErbB2 gene (erbB2, also known as her2, or c-erbB-2), which encodes a 185-kd transmembrane glycoprotein receptor
`(p185HER2) related to the epidermal growth factor receptor (EGFR), is overexpressed in about 25% to 30% of human
`breast cancer (Siamon et al., Science 235:177-182 [1987]; Slamon et al., Science 244:707-712 [1989]).
`[0003] Several lines of evidence support a direct role for ErbB2 in the pathogenesis and clinical aggressiveness of
`ErbB2-overexpressing tumors. The introduction of ErbB2 into non-neoplastic cells has been shown to cause their ma(cid:173)
`lignant transformation (Hudziak et al., Proc. Natl. Acad. Sci. USA 84:7159-7163 [1987]; DiFiore et al., Science 237:
`78-182 [1987]). Transgenic mice that express HER2 were found to develop mammary tumors (Guy et al., Proc. Natl.
`Acad. Sci. USA 89:10578-10582 [1992]).
`[0004] Antibodies directed against human erbB2 protein products and proteins encoded by the rat equivalent of the
`erbB2 gene (neu) have been described. Drebin et al., Cell 41:695-706 (1985) refer to an lgG2a monoclonal antibody
`which is directed against the rat neu gene product. This antibody called 7.16.4 causes down-modulation of cell surface
`p185 expression on B 1 04-1-1 cells (N IH-3T3 cells transfected with the neu proto-oncogene) and inhibits colony formation
`of these cells. In Drebin et al. PNAS (USA) 83:9129-9133 (1986), the 7.16.4 antibody was shown to inhibit the tumorigenic
`growth of neu-transformed NIH-3T3 cells as well as rat neuroblastoma cells (from which the neu oncogene was initially
`isolated) implanted into nude mice. Drebin et al. in Oncogene 2:387-394 (1988) discuss the production of a panel of
`antibodies against the rat neu gene product. All of the antibodies were found to exert a cytostatic effect on the growth
`of neu-transformed cells suspended in soft agar. Antibodies of the lgM, lgG2a and lgG2b isotypes were able to mediate
`significant in vitro lysis of neu-transformed cells in the presence of complement, whereas none of the antibodies were
`able to mediate high levels of antibody-dependent cellular cytotoxicity (ADCC) of the neu-transformed cells. Drebin et
`al. Oncogene 2:273-277 (1988) report that mixtures of antibodies reactive with two distinct regions on the p185 molecule
`result in synergistic anti-tumor effects on neu-transformed NIH-3T3 cells implanted into nude mice. Biological effects of
`anti-neu antibodies are reviewed in Myers et al., Meth. Enzym. 198:277-290 (1991 ). See also W094/22478 published
`October 13, 1994.
`[0005] Hudziak et al., Mol. Cell. Bioi. 9(3):1165-1172 (1989) describe the generation of a panel of anti-ErbB2 antibodies
`which were characterized using the human breast tumor cell line SKBR3. Relative cell proliferation of the SKBR3 cells
`following exposure to the antibodies was determined by crystal violet staining of the monolayers after 72 hours. Using
`this assay, maximum inhibition was obtained with the antibody called 4D5 which inhibited cellular proliferation by 56%.
`Other antibodies in the panel, including 7C2 and 7F3, reduced cellular proliferation to a lesser extent in this assay.
`Hudziak eta/. conclude that the effect of the 4D5 antibody on SKBR3 cells was cytostatic rather than cytotoxic, since
`SKBR3 cells resumed growth at a nearly normal rate following removal of the antibody from the medium. The antibody
`4D5 was further found to sensitize p185erbB2-overexpressing breast tumor cell lines to the cytotoxic effects of TN F-a.
`See also W089/06692 published July 27, 1989. The anti-ErbB2 antibodies discussed in Hudziak et a/. are further
`characterized in Fendly et al. Cancer Research 50:1550-1558 (1990); Kotts et al. In Vitro 26(3):59A (1990); Sarup et
`al. Growth Regulation 1 :72-82 (1991 ); Shepard et al. J. Clin. lmmunol. 11 (3):117-127 (1991 ); Kumar et al. Mol. Cell.
`Bioi. 11 (2):979-986 (1991); Lewis et al. Cancer lmmunol. lmmunother. 37:255-263 (1993); Pietras et al. Oncogene 9:
`1829-1838 (1994); Vitetta et al. Cancer Research 54:5301-5309 (1994); Sliwkowski et al. J. Bioi. Chem. 269(20):
`14661-14665 (1994); Scott et al. J. Bioi. Chem. 266:14300-5 (1991 ); and D'souza et al. Proc. Natl. Acad. Sci. 91:
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`7202-7206 (1994).
`[0006] Tagliabue et al. Int. J. Cancer 47:933-937 (1991) describe two antibodies which were selected for their reactivity
`on the lung adenocarcinoma cell line (Calu-3) which overexpresses ErbB2. One of the antibodies, called MGR3, was
`found to internalize, induce phosphorylation of ErbB2, and inhibit tumor cell growth in vitro.
`[0007] McKenzie et al. Oncogene 4:543-548 (1989) generated a panel of anti-ErbB2 antibodies with varying epitope
`specificities, including the antibody designated TA1. This TA1 antibody was found to induce accelerated endocytosis
`of ErbB2 (see Maier et al. Cancer Res. 51:5361-5369 [1991 ]). Bacus et al. Molecular Carcinogenesis 3:350-362 (1990)
`reported that the TA 1 antibody induced maturation of the breast cancer cell lines AU-565 (which overexpresses the
`erb82 gene) and MCF-7 (which does not). Inhibition of growth and acquisition of a mature phenotype in these cells was
`found to be associated with reduced levels of ErbB2 receptor at the cell surface and transient increased levels in the
`cytoplasm.
`[0008] Stancovski et al. PNAS (USA) 88:8691-8695 (1991) generated a panel of anti-ErbB2 antibodies, injected them
`i.p. into nude mice and evaluated their effect on tumor growth of murine fibroblasts transformed by overexpression of
`the erbB2 gene. Various levels of tumor inhibition were detected for four of the antibodies, but one of the antibodies
`(N28) consistently stimulated tumor growth. Monoclonal antibody N28 induced significant phosphorylation of the ErbB2
`receptor, whereas the other four antibodies generally displayed low or no phosphorylation-inducing activity. The effect
`of the anti-ErbB2 antibodies on proliferation of SKBR3 cells was also assessed. In this SKBR3 cell proliferation assay,
`two of the antibodies (N12 and N29) caused a reduction in cell proliferation relative to control. The ability of the various
`antibodies to induce cell lysis in vitro via complement-dependent cytotoxicity (CDC) and antibody-mediated cell-depend-
`ent cytotoxicity (ADCC) was assessed, with the authors of this paper concluding that the inhibitory function of the
`antibodies was not attributed significantly to CDC or ADCC.
`[0009] Bacus et al. Cancer Research 52:2580-2589 (1992) further characterized the antibodies described in Bacus
`eta/. (1990) and Stancovski eta/. of the preceding paragraphs. Extending the i.p. studies of Stancovski eta/., the effect
`of the antibodies after i.v. injection into nude mice harboring mouse fibroblasts overexpressing human ErbB2 was
`assessed. As observed in their earlier work, N28 accelerated tumor growth, whereas N12 and N29 significantly inhibited
`growth of the ErbB2-expressing cells. Partial tumor inhibition was also observed with the N24 antibody. Bacus eta/.
`also tested the ability of the antibodies to promote a mature phenotype in the human breast cancer cell lines AU-565
`and MDA-MB453 (which overexpress ErbB2) as well as MCF-7 (containing low levels of the receptor). Bacus eta/. saw
`a correlation between tumor inhibition in vivo and cellular differentiation; the tumor-stimulatory antibody N28 had no
`effect on differentiation, and the tumor inhibitory action of the N12, N29 and N24 antibodies correlated with the extent
`of differentiation they induced.
`[001 0] Xu et al. Int. J. Cancer 53:401-408 (1993) evaluated a panel of anti-ErbB2 antibodies for their epitope binding
`specificities, as well as their ability to inhibit anchorage-independent and anchorage-dependent growth of SKBR3 cells
`(by individual antibodies and in combinations), modulate cell-surface ErbB2, and inhibit ligand stimulated anchorage-
`independent growth. See also W094/00136 published Jan 6, 1994 and Kasprzyk et al. Cancer Research 52:2771-2776
`(1992) concerning anti-ErbB2 antibody combinations. Other anti-ErbB2 antibodies are discussed in Hancock et al.
`Cancer Res.51:4575-4580(1991); Shawver et al. Cancer Res. 54:1367-1373 (1994); Arteaga et al. Cancer Res. 54:
`3758-3765 (1994); and Harwerth et al. J. Bioi. Chern. 267:15160-15167 (1992).
`[0011] A recombinant humanized anti-ErbB2 monoclonal antibody (a humanized version of the murine anti-ErbB2
`antibody 4D5, referred to as rhuMAb HER2, HERCEPTIN®, or HERCEPTIN® anti-ErbB2 antibody) has been clinically
`active in patients with ErbB2-overexpressing metastatic breast cancers that had received extensive prior anti-cancer
`therapy (Baselga et al., J. Clin. Oncol. 14:737-744 [1996]). The recommended initial loading dose for HERCEPTIN® is
`4mg/kg administered as a 90-minute infusion. The recommended weekly maintenance dose is 2mg/kg and can be
`administered as a 30-minute infusion if the initial loading dose is well tolerated.
`[0012] ErbB2 overexpression is commonly regarded as a predictor of a poor prognosis, especially in patients with
`primary disease that involves axillary lymph nodes (Siamon eta/., [1987] and [1989], supra; Ravdin and Chamness,
`Gene 159:19-27 [1995]; and Hynes and Stern, Biochim Biophys Acta 1198:165-184 [1994]), and has been linked to
`sensitivity and/or resistance to hormone therapy and chemotherapeutic regimens, including CMF (cyclophosphamide,
`methotrexate, and fluoruracil) and anthracyclines (Baselga et al., Oncology 11 (3 Suppl 1 ):43-48 [1997]). However,
`despite the association of ErbB2 overexpression with poor prognosis, the odds of HER2-positive patients responding
`clinically to treatment with taxanes were greater than three times those of HER2-negative patients (Ibid). rhuMab HER2
`was shown to enhance the activity of paclitaxel (TAXOL®) and doxorubicin against breast cancer xenografts in nude
`mice injected with BT-474 human breast adenocarcinoma cells, which express high levels of HER2 (Baselga et al.,
`Breast Cancer, Proceedings of ASCO, Vol. 13, Abstract 53 [1994]).
`
`Summa of the Invention
`
`[0013] The present disclosure concerns the discovery that an early attainment of an efficacious target trough serum
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`concentration by providing an initial dose or doses of anti-ErbB2 antibodies followed by subsequent doses of equal or
`smaller amounts of antibody (greater front loading) is more efficacious than conventional treatments. The efficacious
`target trough serum concentration is reached in 4 weeks or less, preferably 3 weeks or less, more preferably 2 weeks
`or less, and most preferably 1 week or less, including 1 day or less. The target serum concentration is thereafter
`maintained by the administration of maintenance doses of equal or smaller amounts for the remainder of the treatment
`regimen or until suppression of disease symptoms is achieved.
`[0014] The disclosure further concerns a method for the treatment of a human patient susceptible to or diagnosed
`with a disorder characterized by overexpression of ErbB2 receptor comprising administering a therapeutically effective
`amount of an anti-ErbB2 antibody subcutaneously. Preferably, the initial dose (or doses) as well as the subsequent
`10 maintenance dose or doses are administered subcutaneously. Optionally, where the patient's tolerance to the anti(cid:173)
`ErbB2 antibody is unknown, the initial dose is administered by intravenous infusion, followed by subcutaneous admin(cid:173)
`istration of the maintenance doses if the patient's tolerance for the antibody is acceptable.
`[0015] According to the disclosure, the method of treatment involves administration of an initial dose of anti-ErbB2
`antibody of more than approximately 4 mg/kg, preferably more than approximately 5 mg/kg. The maximum initial dose
`or a subsequent dose does not exceed 50 mg/kg, preferably does not exceed 40 mg/kg, and more preferably does not
`exceed 30 mg/kg. Administration is by intravenous or subcutaneous administration, preferably intravenous infusion or
`bolus injection, or more preferably subcutaneous bolus injection. The initial dose may be one or more administrations
`of drug sufficient to reach the target trough serum concentration in 4 weeks or less, preferably 3 weeks or less, more
`preferably 2 weeks or less, and most preferably 1 week or less, including one day or less.
`[0016] According to the disclosure, the initial dose or doses is/are followed by subsequent doses of equal or smaller
`amounts of antibody at intervals sufficiently close to maintain the trough serum concentration of antibody at or above
`an efficacious target level. Preferably, an initial dose or subsequent dose does not exceed 50 mg/kg, and each subsequent
`dose is at least 0.01 mg/kg. Preferably the amount of drug administered is sufficient to maintain the target trough serum
`concentration such that the interval between administration cycles is at least one week. Preferably the trough serum
`concentration does not exceed 2500 f-lg/ml and does not fall below 0.01 f-lg/ml during treatment. The front loading drug
`treatment method of the disclosure has the advantage of increased efficacy by reaching a target serum drug concentration
`early in treatment. The subcutaneous delivery of maintenance doses according to the disclosure has the advantage of
`being convenient for the patient and health care professionals, reducing time and costs for drug treatment. Preferably,
`the initial dose (or the last dose within an initial dose series) is separated in time from the first subsequent dose by 4
`weeks or less, preferably 3 weeks or less, more preferably 3 weeks or less, most preferably 1 week or less.
`In an embodiment, the initial dose of anti-ErbB2 is 6 mglkg, 8 mg/kg, or 12 mg/kg delivered by intravenous or
`[0017]
`subcutaneous administration, such as intravenous infusion or subcutaneous bolus injection. The subsequent mainte(cid:173)
`nance doses are 2 mg/kg delivered once per week by intravenous infusion, intravenous bolus injection, subcutaneous
`infusion, or subcutaneous bolus injection. The choice of delivery method for the initial and maintenance doses is made
`according to the ability of the animal or human patient to tolerate introduction of the antibody into the body. Where the
`antibody is well-tolerated, the time of infusion may be reduced. The choice of delivery method as disclosed for this
`embodiment applies to all drug delivery regimens contemplated according to the invention.
`In another embodiment, the disclosure includes an initial dose of 12 mg/kg anti-ErbB2 antibody, followed by
`[0018]
`subsequent maintenance doses of 6 mg/kg once per 3 weeks.
`In still another embodiment, the disclosure includes an initial dose of 8 mg/kg anti-ErbB2 antibody, followed
`[0019]
`by 6 mg/kg once per 3 weeks.
`In yet another embodiment, the disclosure includes an initial dose of 8 mg/kg anti-ErbB2 antibody, followed by
`[0020]
`subsequent maintenance doses of 8 mg/kg once per week or 8 mg/kg once every 2 to 3 weeks.
`In another embodiment, the disclosure includes initial doses of at least 1 mg/kg, preferably 4 mg/kg, anti-ErbB2
`[0021]
`antibody on each of days 1, 2 and 3, followed by subsequent maintenance doses of 6 mg/kg once per 3 weeks.
`In another embodiment, the disclosure includes an initial dose of 4 mg/kg anti-ErbB2 antibody, followed by
`[0022]
`subsequent maintenance doses of 2 mg/kg twice per week, wherein the maintenance doses are separated by 3 days.
`In still another embodiment, the disclosure includes a cycle of dosing in which delivery of anti-ErbB2 antibody
`[0023]
`is 2-3 times per week for 3 weeks. In one embodiment, each dose is approximately 25 mg/kg or less for a human patient,
`preferably approximately 10 mglkg or less. This 3 week cycle is preferably repeated as necessary to achieve suppression
`of disease symptoms.
`In another embodiment, the disclosure includes a cycle of dosing in which delivery of anti-ErbB2 antibody is
`[0024]
`daily for 5 days. According to the disclosure, the cycle is preferably repeated as necessary to achieve suppression of
`disease symptoms.
`[0025] The disorder preferably is a benign or malignant tumor characterized by the overexpression of the ErbB2
`receptor, e.g. a cancer, such as, breast cancer, squamous cell cancer, small-cell lung cancer, non-small cell lung cancer,
`gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer,
`hepatoma, colon cancer, colo rectal cancer, endometrial carcinoma, salivary gland carcinoma, kidney cancer, liver cancer,
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`prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma and various types of head and neck cancer. The
`method of the invention may further comprise administration of a chemotherapeutic agent other than an anthracycline,
`e.g. doxorubicin or epirubicin. The chemotherapeutic agent preferably is a taxoid, such as TAXOL® (paclitaxel) or a
`TAXOL® derivative.
`[0026] Preferred anti-ErbB2 antibodies bind the extracellular domain of the ErbB2 receptor, and preferably bind to
`the epitope 405 or 3H4 within the ErbB2 extracellular domain sequence. More preferably, the antibody is the antibody
`405, most preferably in a humanized form. Other preferred ErbB2-binding antibodies include, but are not limited to,
`antibodies 7C2, 7F3, and 2C4, preferably in a humanized form.
`[0027] The method of the present disclosure is particularly suitable for the treatment of breast or ovarian cancer,
`characterized by the overexpression of the ErbB2 receptor.
`[0028] The present application also provides a method of therapy involving infrequent dosing of an anti-ErbB2 antibody.
`In particular, the disclosure provides a method for the treatment of cancer (e.g. cancer characterized by overexpression
`of the ErbB2 receptor) in a human patient comprising administering to the patient a first dose of an anti-ErbB2 antibody
`followed by at least one subsequent dose of the antibody, wherein the first dose and subsequent dose are separated
`from each other in time by at least about two weeks (e.g. from about two weeks to about two months), and optionally
`at least about three weeks (e.g. from about three weeks to about six weeks). For instance, the antibody may be admin(cid:173)
`istered about every three weeks, about two to about 20 times, e.g. about six times. The first dose and subsequent dose
`may each be from about 2mg/kg to about 16mg/kg; e.g. from about 4mg/kg to about 12mg/kg; and optionally from about
`6mg/kg to about 12mg/kg. Generally, two or more subsequent doses (e.g. from about two to about ten subsequent
`doses) of the antibody are administered to the patient, and those subsequent doses are preferably separated from each
`other in time by at least about two weeks (e.g. from about two weeks to about two months), and optionally at least about
`three weeks (e.g. from about three weeks to about six weeks). The two or more subsequent doses may each be from
`about 2mg/kg to about 16mg/kg; or from about 4mg/kg to about 12mg/kg; or from about 6mg/kg to about 12mg/kg. The
`disclosure additionally provides an article of manufacture, comprising a container, a composition within the container
`comprising an anti-ErbB2 antibody, and a package insert containing instructions to administer the antibody according
`to such methods.
`[0029] The presently described dosing protocols may be applied to other anti-ErbB antibodies such as anti-epidermal
`growth factor receptor (EG FR), anti-ErbB3 and anti-ErbB4 antibodies. Thus, the disclosure provides a method for the
`treatment of cancer in a human patient, comprising administering an effective amount of an anti-ErbB antibody to the
`human patient, the method comprising administering to the patient an initial dose of at least approximately 5 mg/kg of
`the anti-ErbB antibody; and administering to the patient a plurality of subsequent doses of the antibody in an amount
`that is approximately the same or less than the initial dose. Alternatively, or additionally, the disclosure pertains to a
`method for the treatment of cancer in a human patient comprising administering to the patient a first dose of an anti(cid:173)
`ErbB antibody followed by at least one subsequent dose of the antibody, wherein the first dose and subsequent dose
`are separated from each other in time by at least about two weeks. The disclosure additionally provides an article of
`manufacture, comprising a container, a composition within the container comprising an anti-ErbB antibody, and a package
`insert containing instructions to administer the antibody according to such methods.
`In another aspect, the disclosure concerns an article of manufacture, comprising a container, a composition
`[0030]
`within the container comprising an anti-ErbB2 antibody, optionally a label on or associated with the container that
`indicates that the composition can be used for treating a condition characterized by overexpression of ErbB2 receptor,
`and a package insert containing instructions to avoid the use of anthracycline-type chemotherapeutics in combination
`with the composition. According to the disclosure, the package insert further includes instructions to administer the anti(cid:173)
`ErbB2 antibody at an initial dose of 5 mg/kg followed by the same or smaller subsequent dose or doses. In another
`embodiment of the invention, the package insert further includes instructions to administer the anti-ErbB2 antibody
`subcutaneously for at least one of the doses, preferably for all of the subsequent doses following the initial dose, most
`preferably for all doses.
`In a further aspect, the disclosure provides a method of treating ErbB2 expressing cancer in a human patient
`[0031]
`comprising administering to the patient effective amounts of an anti-ErbB2 antibody and a chemotherapeutic agent. In
`one embodiment of the disclosure, the chemotherapeutic agent is a taxoid including, but not limited to, paclitaxel and
`docetaxel. In another embodiment, the chemotherapeutic agent is an anthracyline derivative including, but not limited
`to, doxorubicin or epirubicin. In still another embodiment, treatment with an anti-ErbB2 antibody and an anthracycline
`derivative further includes administration of a cardioprotectantto the patient. In still another embodiment, an anthracycline
`derivative is not administered to the patient with the anti-ErbB2 antibody. One or more additional chemotherapeutic
`agents may also be administered to the patient. The cancer is preferably characterized by overexpression of ErbB2.
`[0032] The disclosure further provides an article of manufacture comprising a container, a composition within the
`container comprising an anti-ErbB2 antibody and a package insert instructing the user of the composition to administer
`the anti-ErbB2 antibody composition and a chemotherapeutic agent to a patient. In another embodiment, the chemo(cid:173)
`therapeutic agent is other than an anthracycline, and is preferably a taxoid, such as TAXOL ®. In still another embodiment,
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`EP 1 210115 81
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`the chemotherapeutic agent is an anthracycline, including but not limited to, doxorubicin or epirubicin. In yet another
`embodiment, the chemotherapeutic agent is an anthracycline and the package insert further instructs the user to ad(cid:173)
`minister a cardioprotectant.
`[0033] The methods and composition of the disclosure comprise an anti-ErbB2 antibody and include a humanized
`anti-ErbB2 antibody. Thus, the disclosure further pertains to a composition comprising an antibody that binds ErbB2
`and the use of the antibody for treating ErbB2 expressing cancer, e.g., ErbB2 overexpressing cancer, in a human. The
`disclosure also pertains to the use of the antibody for treating EGFR expressing cancer. Preferably the antibody is a
`monoclonal antibody 4D5, e.g., humanized 4D5 (and preferably huMAb4D5-8 (HERCEPTIN® anti-ErbB2 antibody); or
`monoclonal antibody 2C4, e.g., humanized 2C4. The antibody may be an intact antibody (e.g., an intact lgG, antibody)
`or an antibody fragment (e.g., a Fab, F(abh, diabody, and the like). The variable light chain and variable heavy chain
`regions of humanized anti-ErbB2 antibody 2C4 are shown in Figs. 5A and 58. The present invention is defined by the
`appended claims.
`
`Brief Description of the Drawings
`
`[0034]
`
`Fig. 1 shows epitope-mapping of the extracellular domain of ErbB2 as determined by truncation mutant analysis
`and site-directed mutagenesis (Nakamura et al. J. of Virology 67(1 0):6179-6191 [Oct 1993]; Renz et al. J. Cell Bioi.
`125(6): 1395-1406 [Jun 1994]). The anti-proliferative MAbs 4D5 and 3H4 bind adjacent to the transmembrane do(cid:173)
`main. The various ErbB2-ECD truncations or point mutations were prepared from eDNA using polymerase chain
`reaction technology. The ErbB2 mutants were expressed as gD fusion proteins in a mammalian expression plasmid.
`This expression plasmid uses the cytomegalovirus promoter/enhancer with SV40 termination and polyadenylation
`signals located downstream of the inserted eDNA. Plasmid DNA was transfected into 293S cells. One day following
`transfection, the cells were metabolically labeled overnight in methionine and cysteine-free, low glucose DMEM
`containing 1% dialyzed fetal bovine serum and 25 fLCi each of 35S methionine and 35S cysteine. Supernatants were
`harvested either the ErbB2 MAbs or control antibodies were added to the supernatant and incubated 2-4 hours at
`4'C. The complexes were precipitated, applied to a 10-20% Tricine SDS gradient gel and electrophoresed at 100
`V. The gel was electroblotted onto a membrane and analyzed by autoradiography. SEQ ID NOs:8 and 9 depict the
`3H4 and 4D5 epitopes, respectively.
`Fig. 2 depicts with underlining the amino acid sequence of Domain 1 of ErbB2 (SEQ ID N0:1 ). Bold amino acids
`indicate the location of the epitope recognized by MAbs 7C2 and 7F3 as determined by deletion mapping, i.e. the
`"7C2/7F3 epitope" (SEQ ID N0:2).
`Fig. 3 is a graph of anti-ErbB2 antibody (HERCEPITN®) trough serum concentration (fLg/ml, mean ± SE, dark
`circles) by week from week 2 through week 36 for ErbB2 overexpressing patients treated with HERCEPTIN® anti(cid:173)
`ErbB2 antibody at 4 mg/kg initial dose, followed by 2 mg/kg weekly. The number of patients at each time point is
`represented by "n" (white squares).
`Fig. 4A is a linear plot of tumor volume changes over time in mice treated with HERCEPTIN® anti-ErbB2 antibody.
`Fig. 48 is a semi-logarithmic plot of the same data as in Fig. 4A such that the variation in tumor volume for the
`treated animals is observed more readily.
`Figs. 5A and 58 depict alignments of the amino acid sequences of the variable light (VL) (Fig. 5A) and variable
`heavy (VH) (Fig. 58) domains of murine monoclonal antibody 2C4 (SEQ ID Nos. 10 and 11, respectively); VL and
`VH domains of humanized Fab version 574 (SEQ ID Nos. 12 and 13, respectively), and human VL and VH consensus
`frameworks (hum Kl, light kappa subgroup I; humlll, heavy subgroup Ill) (SEQ ID Nos. 14 and 15, respectively).
`Asterisks identify differences between humanized Fab version 574 and murine monoclonal antibody 2C4 or between
`humanized Fab version 574 and the human framework. Complementarity Determining Regions (CDRs) are in
`brackets. Humanized Fab version