(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY(PCT)
`
`(19) World Intellectual Property Organization
`International Bureau
`
`(43) International Publication Date
`
`18 October 2007 (18.10.2007)
`
`(10) International Publication Number
`WO 2007/118214 A2
`
`Agents: WOODCOCK WASHBURN LLPet al.; Cira
`Centre, 12th Floor, 2929 Arch Street, Philadelphia, Penn-
`sylvania 19104-2891 (US).
`
`Designated States (unless otherwise indicated, for every
`kind of national protection available): AE, AG, AL, AM,
`AT, AU, AZ, BA, BB, BG, BH, BR, BW,BY, BZ, CA, CH,
`CN, CO, CR, CU, CZ, DE, DK, DM, DZ, EC, EE, EG, ES,
`FI, GB, GD, GE, GH, GM, GT, HN, HR, HU,ID,IL, IN,
`TS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC, LK, LR,
`LS, LT, LU, LY, MA, MD, MG, MK, MN, MW,MX, MY,
`MZ, NA, NG, NI, NO, NZ, OM, PG, PH, PL, PT, RO, RS,
`RU, SC, SD, SE, SG, SK, STL, SM, SV, SY, TJ, T, TN,
`TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW.
`
`(51) International Patent Classification:
`C07K 16/22 (2006.01)
`A6LK 39/395 (2006.01)
`GOIN 33/53 (2006.01)
`A6IP 35/00 (2006.01)
`C1I2N 15/13 (2006.01)
`A61K 51/10 (2006.01)
`CI2N 5/10 (2006.01)
`
`(74)
`
`(81)
`
`(21) International Application Number:
`PCT/US2007/066180
`
`(22) International Filing Date:
`
`6 April 2007 (06.04.2007)
`
`(25) Filing Language:
`
`(26) Publication Language:
`
`(30) Priority Data:
`60/790,512
`
`English
`
`English
`
`7 April 2006 (07.04.2006)
`
`US
`
`(71) Applicant (for all designated States except US): THE
`GOVERNMENT OF THE UNITED STATES OF
`AMERICA AS REPRESENTED BY THE SECRE-
`TARY, DEPARTMENT OF HEALTH AND HUMAN
`SERVICES [US/US]; 6011 Executive Blvd., Suite 325,
`Rockville, Maryland 20852-3804 (US).
`
`(72) Inventors; and
`(for US only): DIMITROV,
`(75) Inventors/Applicants
`Dimiter S. [US/US]; 1741 Northridge Lane, Frederick,
`Maryland 21702 (US). ZHU, Zhongyu [CN/US]; 2501
`Carrington Way, Frederick, Maryland 21702 (US).
`
`(84)
`
`Designated States (unless otherwise indicated, for every
`kind of regional protection available): ARIPO (BW, GH,
`GM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM,
`ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM),
`European (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI,
`FR, GB, GR,HU,IE,IS, IT, LT, LU, LV, MC, MT, NL, PL,
`PT, RO,SE, SI, SK, TR), OAPI (BF, BJ, CF, CG, CI, CM,
`GA, GN, GQ, GW, ML, MR, NE, SN, TD, TG).
`Published:
`
`without international search report and to be republished
`uponreceipt of that report
`
`For two-letter codes and other abbreviations, refer to the "Guid-
`ance Notes on Codes and Abbreviations” appearing at the begin-
`ning of each regular issue of the PCT Gazette.
`
`(54) Titles ANTIBODY COMPOSITIONS AND METHODS FOR TREATMENTOF NEOPLASTIC DISEASE
`
`oeIGFl4m708.2-
`£<
`a6[GFl+m708
`
`Sl
`
`O + o
`
`lO
`
`
`
`WO2007/118214A2INTIMATIONTMITUAAA
`
`Antibody Concentration
`
`~~ IGFlK4-m606
`
`IGF ll-m708.2
`
`-+ IGFlem708
`
`(57) Abstract: Antibody compositions and methodsfor treatmentof neoplastic disease in a mammalian subject are provided. Meth-
`ods of diagnosing cancer in a mammalian subject are also provided.
`
`

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`WO 2007/118214
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`PCT/US2007/066180
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`ANTIBODY COMPOSITIONS AND METHODS FOR TREATMENT
`OF NEOPLASTIC DISEASE
`
`CROSS REFERENCE TO RELATED APPLICATION
`
`[0001]
`
`This application claimspriority to U.S. Provisional Application No. 60/790,512,
`
`filed April 7, 2006, the disclosure of which is incorporated by reference in its entirety.
`
`FIELD
`
`[0002] The invention generally relates to an antibody composition and a methodfor
`
`treatment of neoplastic disease in a mammalian subject. An isolated human monoclonal
`
`antibody binds to insulin-like growth factor | or to both insulin-like growth factor | and insulin-
`
`like growth factor II. The invention further relates to methods of diagnosing cancer in a
`
`mammalian subject.
`
`BACKGROUND
`
`[0003] Cancer therapies are based on the theory that cells with accelerated rates of
`
`division andproliferation are predisposed to the development of cancer. Recently, a numberof
`
`epidemiologic studies have shown consistently that high circulating levels of a potent mitogen,
`
`insulin-like growth factor (GF)-I, are associated with increased risk for several commoncancers,
`
`including those of the breast, prostate, lung, and colorectum. The level of IGF-binding protcin
`
`(IGFBP)-3, a major IGF-I-binding protein in serum that, in most situations, suppresses the
`
`mitogenic action of IGF-I, is inversely associated with the risk of these cancers.
`
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`WO 2007/118214
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`[0004]
`
`Functionally, IGF-I not only stimulates cell proliferation but also inhibits
`
`apoptosis. The combination of these mitogenic and antiapoptotic effects can have an impact on
`
`tumor growth. Besides their direct effect on cancer-related cellular activities, members of the IGF
`
`family also interact with a variety of molecules that are involved in cancer development and
`
`progression, including the sex steroid hormones, products of tumor suppressor genes, and other
`
`growth factors. Furthermore, the expression and production of IGF-I, a peptide hormonethatis
`
`involved in regulating human growth and development, are influenced by nutrition and physical
`
`activity. Experiments to understand the molecular structure and physiologic function of
`
`members of the IGF family provide insights into the role ofmitogenic growth factors in
`
`carcinogenesis. Yu and Rohan, /. Natl. Cancer Inst. 92: 1472-1489, 2000.
`
`[0005]
`
`IGFs stimulate the proliferation of cultured humanbreast cancer cells. This
`
`stimulation is mediated through the receptor, insulin-like growth factor receptor 1 (IGFR-1),
`
`which is a memberof the receptor tyrosine kinase family. Whenactivated byits ligands (IGF-I
`
`or IGF-II, IGFR1 phosphorylates tyrosine residues on two major substrates, IRS-1 and Shc,
`
`which subsequently signal through the Ras/Raf and phosphatidylinositol 3'-kinase/AKT
`
`pathways. IGFR1 plays a crucial role in transformation. Cells derived from IGFR1 knockout
`
`mice are resistant to transformation by various viral and cellular oncogenes, including SV40
`
`large T antigen and activated ras, whereasfibroblast cells from wild-type mice can be readily
`
`transformed by these oncogenes.
`
`[0006] There is increasing epidemiological evidence to link elevated plasma IGF-I level
`
`with prostate, breast, and colon cancer risk. Breast cancer tissues from patients exhibit higher
`
`IGFR1 expression than adjacent normal tissue, suggesting a link between IGFR1 and breast
`
`epithelial cell transformation. It has becn reported that the transformation capacity oftumorcclls
`
`is attenuated when IGFR1 is inhibited using an antisensestrategy, neutralizing antibody (anti-IR3
`
`or anti-IGF-I) or dominant negative truncation of the receptor. Hailey, J. et al, Molecular Cancer
`
`Therapeutics 1: 1349-1353, 2002; Maloney E.K., et al, Cancer Res. 63: 5073-5083, 2003;
`
`Burtrum D., et al, Cancer Res., 63: 8912-8921, 2003; Luetal., J. Biol. Chem. 279: 2856-2865,
`
`2004; Miyamotoet al., Clin. Cancer Res. 11: 3494-3502, 2005; Goya et al., Cancer Research
`
`64: 6252-6258, 2004. A needexists in the art for improved multi-target therapies to treat
`
`neoplastic disease and metastatic cancers.
`
`SUMMARY
`
`[0007] The present invention generally relates to antibody compositions and methods
`
`for treatment of neoplastic disease in a mammalian subject. The present invention further relates
`
`to methods of diagnosing neoplastic disease in a mammalian subject. The antibody compositions
`-2-
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`are isolated monoclonal antibodies that bind to insulin-like growth factor I. Another set of
`
`antibody compositions are monoclonal antibodies that bind to insulin-like growth factor I and are
`
`cross-reactive to and bind to insulin-like growth factor II. The isolated monoclonal antibodies
`
`are, for example, human, non-humanprimate, rabbit, rat or mouse antibodies. Theisolated
`
`human monoclonal antibody compositionsthat bind to insulin-like growth factorI are, for
`
`example, m705 and m706. The isolated human monoclonal antibody compositions that bind to
`
`both insulin-like growth factor I and insulin-like growth factor Hl are m708 and m708.2.
`
`Monoclonal antibodies m705, m706, m708, and m708.2 do not bind to human insulin. m705 has
`
`a Vy chain amino acid sequence comprising SEQ ID NO: | and a Vy; chain amino acid sequence
`
`comprising SEQ ID NO: 2. m706 has a Vy chain amino acid sequence comprising SEQ ID NO:
`
`3 and a V, chain amino acid sequence comprising SEQ ID NO: 4. m708 has a Vy chain amino
`
`acid sequence comprising SEQ ID NO: 5 and a V_, chain amino acid sequence comprising SEQ
`
`ID NO: 6. m708.2 has a Vy chain amino acid sequence comprising SEQ ID NO: 7 and a Vr.
`
`chain amino acid sequence comprising SEQ ID NO: 8.
`
`[0008] An isolated monoclonal antibody is provided which binds to human insulin-like
`
`growth factor I and human insulin-like growth factor IT comprising an amino acid sequencein its
`
`heavy chain variable region as set forth in SEQ ID NO: 7 or an amino acid sequence whichis at
`
`least 90% homologous to SEQ ID NO: 7.
`
`[0009] An isolated monoclonal antibody is provided which binds to human insulin-like
`
`growth factor I and humaninsulin-like growth factor II comprising an amino acid sequenceinits
`
`light chain variable region as set forth in SEQ ID NO: 8 or an amino acid sequence whichis at
`
`least 90% homologous to SEQ ID NO: 8.
`
`[0010] Anisolated monoclonal antibody is provided which binds to human insulin-like
`
`growth factor | and humaninsulin-like growth factor Il comprising amino acid sequencesin their
`
`heavy chain variable regionsor light chain variable regions as set forth in SEQ ID NOs: 7 and 8,
`
`respectively, or amino acid sequences whichareat least 90% homologous, respectively. A
`
`pharmaceutical composition is provided comprising one or more of the antibodies of the present
`
`invention and a pharmaceutically acceptable carrier.
`
`[0011]
`
`Ina further aspect, the antibody provides at least one CDR sequenceincluding,
`
`but not limited to, Vr: QS IS S (SEQ ID NO: 9), Vi: AAS (SEQ ID NO: 10), Wr: QQSYS
`
`TPS TF(SEQIDNO: 11), Va: GG TFSS Y A(SEQIDNO: 12), Vo: GIIPILGIA
`
`(SEQ ID NO: 13), or Vu; ARGPRGYSYNFED/Y (SEQ ID NO: 14). In a further aspect,
`
`the antibody includes, but is not limited to, an IgG), an [gGp, an IgGs, an IgGy, an IgM, an IgAj,
`
`an IgA», a secretory IgA, an IgD, or an IgE antibody. The antibody can be an IgGik or IgG ir
`
`-3-
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`isotype. the antibody is an IgG4x or IgG4A isotype. The antibody can be an IgG, an IgGo, an
`
`IgGs, an IgG4, an IgM, an IgA, an IgAo, a secretory IgA, an IgD, or an IgE antibody. The
`
`antibody can be an IgGik or IgG)A isotype. The antibody can be an IgG4xk or IgG4A isotype. In a
`
`detailed aspect the antibody is human, non-humanprimate, rabbit, rodent, rat, or mouse, or a
`
`combinationthereof.
`
`[0012]
`
`In another aspect, the isolated monoclonalantibody of the present invention has
`
`one or more of the following characteristics: (1) inhibits IGF-1 receptor phosphorylation in an in
`
`vitro MCF-7 breast cancer cell assay at an antibody concentration about 4 nM orgreater; (11)
`
`inhibits IGF-I binding or IGF-II binding to IGF-1 receptor; or (111) inhibits cell migration in a cell
`
`migration assay.
`
`In another aspect, the isolated monoclonalantibody of the present invention has
`[0013]
`a dissociation equilibrium constant (Kp) of approximately 10° M or less, when determined by
`
`surface plasmon resonance (SPR) using recombinant human insulin-like growth factor I or
`
`humaninsulin-like growth factor II as an analyte and the antibody as a ligand.
`
`[0014] An isolated monoclonal antibody is provided in a further aspect whichis
`
`capable of binding human insulin-like growth factor I and insulin-like growth factor IT with a
`bindingaffinity of about 10° M"or greater. An isolated monoclonal antibody is provided in a
`
`further aspect which is capable of binding humaninsulin-like growth factor I and insulin-like
`growth factor II with a binding affinity of about 10° M"or greater. In a detailed aspect, the
`
`isolated monoclonal antibody is an intact antibody, an intact IgG, antibody, an intact IgG»
`
`antibody, an intact IgG; antibody, an intact IgG, antibody, an intact IgM antibody, an intact IgA,
`
`antibody, an intact IgA» antibody, an intact secretory IgA antibody, an intact IgD antibody, or an
`
`intact IgE antibody, wherein the antibody is glycosylated in a eukaryotic cell. In a further aspect,
`
`the isolated monoclonal antibody is an antibody fragment or a single chain antibody. The
`
`antibody can be a monoclonal antibody. The antibody can be a F(ab’)», Fab, Fv, or Fd fragment.
`
`The antibody can be antigen-specific.
`
`[0015]
`
`Ina further aspect, the isolated monoclonal antibody of the present invention is
`
`a binding-domain immunoglobulin fusion protein comprising (i) a variable heavy chain amino
`
`acid sequenceas set forth in SEQ ID NO: 7 or a variable heavy chain sequence whichisat least
`
`90% homologous to SEQ ID NO: 7, fused to a variable light chain amino acid sequenceasset
`
`forth in SEQ ID NO: 8 or a variable light chain sequence whichis at least 90% homologousto
`
`SEQ ID NO: 8 via a linker peptide, that is fused to an inmunoglobulin hinge region polypeptide,
`
`(i) an immunoglobulin heavy chain CH2 constant region fused to the hinge region, and(iii) an
`
`immunoglobulin heavy chain CH3 constant region fused to the CH2 constant region. The
`
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`antibody can bind to a predetermined antigen with an equilibrium association constant (Ka), for
`example,of at least 10° M', of at least 10’ M"', or of at least 10'° M"'.
`
`[0016] Anisolated human monoclonal antibody is provided which binds to human
`
`insulin-like growth factor I and humaninsulin-like growth factor II. In one aspect, the antibody
`
`comprises at least one CDR sequence of: Vt: QS IS S (SEQ ID NO: 9), Vi: A A S (SEQ ID
`
`NO: 10), Vr: QQSYSTPS TF(SEQIDNO: 11), Ve: GGTFSS Y A (SEQ ID NO: 12),
`
`Vu: GILPIL GIA (SEQIDNO: 13), or Vu: ARGPRGYSYNFEDY GEQID NO: 14).
`
`[0017] An isolated human monoclonal antibody is provided which binds to human
`
`insulin-like growth factor I comprising an amino acid sequence in its human heavy chain
`
`variable region as set forth in SEQ ID NO: 1 or an amino acid sequence whichis at least 90%
`
`homologous to SEQ ID NO: 1. An isolated human monoclonal antibody is provided which binds
`
`to humaninsulin-like growth factor I comprising an amino acid sequence in its human light
`
`chain variable region as set forth in SEQ ID NO: 2 or an amino acid sequence whichis at least
`
`90% homologous to SEQ ID NO: 2. An isolated human monoclonal antibody which binds to
`
`humaninsulin-like growth factor | comprising an amino acid sequencein its human heavy chain
`
`variable region as set forth in SEQ ID NO: 3 or an amino acid sequence whichis at least 90%
`
`homologous to SEQ ID NO: 3. An isolated human monoclonal antibody is provided which binds
`
`to human insulin-like growth factor I comprising an amino acid sequence in its humanlight
`
`chain variable region as set forth in SEQ ID NO: 4 or an amino acid sequence whichisat least
`
`90% homologous to SEQ ID NO: 4. An isolated human monoclonal antibody is provided which
`
`binds to human insulin-like growth factor I and human insulin-like growth factor II comprising
`
`an amino acid sequence in its human heavy chain variable region as set forth in SEQ ID NO: 5 or
`
`an amino acid sequence whichis at least 90% homologous to SEQ ID NO: 5. An isolated human
`
`monoclonal antibody is provided which binds to human insulin-like growth factor | and human
`
`insulin-like growth factor II comprising an amino acid sequence in its human light chain variable
`
`region as set forth in SEQ ID NO: 6 or an amino acid sequence whichis at least 90%
`
`homologous to SEQ ID NO: 6. A pharmaceutical composition is provided comprising one or
`
`moreof the antibodies of the present invention and a pharmaceutically acceptable carrier.
`
`[0018] An isolated nucleic acid is provided encoding the heavy chain immunoglobulin
`
`variable domain sequenceorthe light chain immunoglobulin variable domain sequence of the
`
`protein/antibody of the present invention. A pharmaceutical composition is provided comprising
`
`the nucleic acid and a pharmaceutically acceptable carrier. A recombinantcell is provided that
`
`contains one or more nucleic acids that encode the immunoglobulin variable domain sequences
`
`of the antibody of the present invention. A host cell that contains a first nucleic acid sequence
`
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`encoding a polypeptide comprising a HC variable domain of an antibody and a second nucleic
`
`acid sequence encoding a polypeptide comprising a LC variable domain ofthe antibody, wherein
`
`the antibody is a protein of the present invention. A method of preparing an antibody capable of
`
`binding insulin growth factor I and insulin growth factor II, the method comprising expressing
`
`the nucleic acid of the present invention in a host cell under conditions to provide for expression
`
`of the nucleic acid, followed by recovery of the antibody.
`
`[0019] An isolated recombinant anti-IGF-I and anti-IGF-L antibody or antigen-binding
`
`fragment thereof, the antibody is provided comprising a human constant region wherein the
`
`antibody or antigen binding fragment (1) competitively inhibits binding of m708.2 antibody
`
`(ATCC Accession No. _) to human IGF-I and human IGF-II, and (11) bindsto a neutralizing
`epitope of human IGF-I and human IGF-II in vivo with an affinity of at least 1 X 10° liter/mole,
`or with an affinity of at least 1 X 10” liter/mole, measured as an associate constant (Ka) as
`
`determined by surface plasmon resonance. The antibody or antigen-binding fragment can
`
`comprise a human constant region and a human variable region. The antibody or antigen-
`
`binding fragment can comprise at least one human light chain and at least one human heavy
`
`chain.
`
`In a further aspect, the light chain comprisesall antigen-binding regions ofthe light chain
`
`of m708.2 (ATCC Accession No. __). The heavy chain can compriseall antigen-binding
`
`regions of the heavy chain of m708.2 (ATCC Accession No. _)._ The light chain can
`
`compriseall antigen-binding regions of the light chain of m708.2 (ATCC Accession No. _)
`
`and wherein the heavy chain comprisesall antigen-binding regions of the heavy chain of m708.2
`
`(ATCC Accession No. __).
`
`[0020] An isolated recombinant anti-IGF-I and anti-IGF-II antibody or antigen-binding
`
`fragment thereof is provided, the antibody comprising a human constant region wherein the
`
`antibody or antigen binding fragment (1) comprises the antigen binding region of m708.2
`
`antibody (ATCC Accession No. __), and (11) binds to a neutralizing epitope of human IGF-I
`and human IGF-II in vivo with an affinity of at least 1 X 10° liter/mole, or with an affinity ofat
`least 1 X 10° liter/mole, measured as an associate constant (Ka) as determined by surface
`
`plasmon resonance. An isolated recombinantanti-IGF-I and anti-IGF-II antibody or antigen-
`
`binding fragment thereof is provided, the antibody comprising a human IgG1 constant region
`
`wherein the antibody or antigen binding fragment (i) competitively inhibits binding of m708.2
`
`antibody (ATCC Accession No. _) to human IGF-I and human IGF-II, and (ii) binds to a
`neutralizing epitope of human IGF-I and human IGF-II in vivo with an affinity of at least 1 X 10°
`liter/mole, or with an affinity of at least 1 X 10° liter/mole, measured as an associate constant
`
`(Ka) as determined by surface plasmon resonance. Anisolated recombinant anti-IGF-I and anti-
`
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`IGF-II antibody or antigen-binding fragment thereof is provided, the antibody comprising a
`
`human IgG1 constant region wherein the antibody or antigen binding fragment (1) comprises the
`
`antigen binding region of m708.2 antibody (ATCC Accession No. __), and (ii) binds to a
`neutralizing epitope of human IGF-I and human IGF-II in vivo with an affinity of at least 1 X 10°
`liter/mole, or with an affinity of at least 1 X 10° liter/mole, measured as an associate constant
`
`(Ka) as determined by surface plasmon resonance.
`
`[0021] A method of detecting human insulin growth factor | and insulin growth factor
`
`II in a sample is provided comprising: (a) providing a sample; (b) contacting the sample of(a)
`
`with a human monoclonal antibody m708 or m708.2 which specifically binds a polypeptide
`
`comprising human insulin growth factor I and insulin growth factor II under conditions which
`
`permit binding of the polypeptide ligand to humaninsulin growth factor I and insulin growth
`
`factor IT; and (c) detecting binding of the antibody m708 or m708.2 with humaninsulin growth
`
`factor I and insulin growth factor II in the sample, wherein detection of binding indicates the
`
`presence of human insulin growth factor I and insulin growth factor II in the sample; thereby
`
`detecting human insulin growth factor | and insulin growth factor I in the sample.
`
`[0022] A method of detecting human insulin growth factor I and insulin growth factor
`
`II in a sample is provided comprising: (a) providing a sample; (b) contacting the sample of(a)
`
`with a human monoclonal antibody m708 or m708.2 whichspecifically binds a polypeptide
`
`comprising human insulin growth factor I and insulin growth factor II under conditions which
`
`permit binding of the polypeptide ligand to human insulin growth factor I and insulin growth
`
`factor II; and (c) detecting binding of the antibody m708 or m708.2 with human insulin growth
`
`factor I and insulin growth factor II in the sample, wherein detection of binding indicates the
`
`presence of human insulin growth factor I and insulin growth factor II in the sample; thereby
`
`detecting human insulin growth factor | and insulin growth factor I in the sample.
`
`[0023] A method of detecting human insulin growth factor I in a sample is provided
`
`comprising: (a) providing a sample; (b) contacting the sample of (a) with a human monoclonal
`
`antibody m705 or m706 whichspecifically binds a polypeptide comprising human insulin
`
`growth factor I under conditions which permit binding of the polypeptide ligand to human
`
`insulin growth factor I; and (c) detecting binding of the antibody m705 or m706 with human
`
`insulin growth factor I in the sample, wherein detection of binding indicates the presence of
`
`human insulin growth factor I in the sample; thereby detecting human insulin growth factor I in
`
`the sample.
`
`[0024] A method of preparing an antibody capable of binding insulin growth factorI
`
`and insulin growthfactor II, the method comprising expressing the nucleic acid of the present
`
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`invention in a host cell under conditions to provide for expression of the nucleic acid, followed
`
`by recovery of the antibody.
`
`[0025] A method ofidentifying a polypeptide ligand specific for human insulin growth
`
`factor I and insulin growth factor II is provided comprising: (a) providing a phage library
`
`comprising phage expressing candidate humaninsulin growth factor I and insulin growth factorI
`
`binding polypeptides; (b) contacting the phage library with human insulin growth factor I and
`
`insulin growth factor LI protein; and (c) detecting binding of the humaninsulin growthfactor |
`
`and insulin growth factor IT protein to phage; thereby identifying a polypeptide ligand specific
`
`for humaninsulin growth factor I and insulin growth factorII.
`
`[0026] A methodfor treating a neoplastic disease in a mammalian subject is provided
`
`comprising administering to the mammalsubject a pharmaceutical composition comprising an
`
`antibody with an amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ
`
`ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8, which specifically
`
`binds to an insulin-like growth factor I in an amount effective to reduce or eliminate the
`
`neoplastic disease in the mammalian subject. In one aspect, the antibody specifically binds to
`
`insulin-like growth factor I and insulin-like growth factor II.
`
`In a further aspect, the antibody
`
`comprises an amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID
`
`NO: 8. The antibody can be linked to a cytotoxic agent. The cytotoxic agent can be a cytotoxic
`
`drug or a radioactive isotope. In a detailed aspect, the neoplastic disease is a solid tumor,
`
`hematological malignancy, leukemia, colorectal cancer, benign or malignant breast cancer,
`
`uterine cancer, uterine leiomyomas, ovarian cancer, endometrial cancer, polycystic ovary
`
`syndrome, endometrial polyps, prostate cancer, prostatic hypertrophy, pituitary cancer,
`
`adenomyosis, adenocarcinomas, meningioma, melanoma, bone cancer, multiple myeloma, CNS
`
`cancer, glioma, or astroblastoma. In a further detailed aspect, the neoplastic disease is tumorcell
`
`metastasis in the mammalian subject. The neoplastic disease can be breast cancer metastasis in
`
`the mammalian subject.
`
`[0027] A method of diagnosing cancer in a mammalian subject suspected of having
`
`neoplastic disease or suspected of being at risk for neoplastic disease is provided comprising,
`
`obtaining a test sample from bloodor tissue of the subject, the test sample comprising a cell
`
`population, providing an antibody comprising an amino acid sequence of SEQ ID NO: 1, SEQ
`
`ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or
`
`SEQ ID NO: 8 to detect the presence or absence of an IGF-I marker on the cells within the cell
`
`population, analyzing the cell population detected by the IGF-I markerto identify and
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`WO 2007/118214
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`PCT/US2007/066180
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`characterize the cells, the presence of IGF-I marker on orin the cells indicative of neoplastic
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`diseaseor risk of neoplastic disease in the mammalian subject.
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`[0028]
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`The method of diagnosing cancer further comprises providing an antibody
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`comprising an amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID
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`NO: 8 to detect the presence or absence of an IGF-II marker and the IGF-I marker on or in the
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`cells within the cell population, and analyzing the ccll population detected by the IGF-I marker
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`and the IGF-I markerto identify and characterize the cells, the presence of the IGF-I marker and
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`the IGF-II marker on orin the cells indicative of neoplastic disease or risk of neoplastic disease
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`in the mammalian subject.
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`[0029]
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`In the diagnostic method, the presence of IGF-I marker or IGF-II marker on or
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`in the cells in the specimen indicates the presence of metastatic cancer in the mammalian subject.
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`In the diagnostic method, the presence of IGF-I marker or IGF-II marker on or in the cells in the
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`specimen indicates the presence of early stage cancer in the mammalian subject. In the
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`diagnostic method, the absence of IGF-I marker and IGF-II marker onorin the cells in the
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`specimenindicates presence of a disease free state or a non-measurable disease state in the
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`mammalian subject.
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`In a further aspect of the diagnostic method, the presence or absence of
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`IGF-I marker or IGF-II marker on or in the cells in the specimen monitors therapy management
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`during cancer therapy or cancer recovery. In a further aspect, the method comprises an imaging
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`moiety associated with the antibody. The imaging moiety can be imaged through magnetic
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`resonance spectroscopy, X-ray spectroscopy, or positron emission tomography (PET). The
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`association can be a covalent bond or a non-covalent bond. The neoplastic disease includes, but
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`is not limited to, solid tumor, hematological malignancy, leukemia, colorectal cancer, breast
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`cancer, uterine cancer, uterine leiomyomas, ovarian cancer, endometrial cancer, polycystic ovary
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`syndrome, endometrial polyps, prostate cancer, prostatic hypertrophy, pituitary cancer,
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`adenomyosis, adenocarcinomas, meningioma, melanoma, bone cancer, multiple myeloma, CNS
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`cancer, glioma,or astroblastoma.
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`[0030] A method of screening a drug candidate compoundfor treatment of cancer in a
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`mammalian subject comprising administering a therapeutically effective amount of the drug
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`candidate compoundto the subject suspected of having cancer, obtaining test samples from
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`blood or tissue of the subject before and after treatment with the drug candidate compound,the
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`test samples comprising a cell population suspected of containing tumorcells, providing an
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`antibody comprising an amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3,
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`SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8 to detect the
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`presence or absence of an IGF-I marker on the cells in the test sample, analyzing the cell
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`population detected by the IGF-I markerto identify the tumorcells in the test samples before
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`treatment with the drug candidate compound comparedto after treatment with the drug candidate
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`compound, wherein the presence of a decreased numberof the tumorcells in the specimen after
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`treatment compared to a numberof the tumorcells in a specimen before treatment indicating
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`effectiveness of the drug candidate compoundin treating the cancer in the mammalian subject.
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`[0031]
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`In another aspect, the mcthod of screening a drug candidate compound for
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`treatment of cancer in a mammalian subject comprises providing an antibody comprising an
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`amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8 to
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`detect the presence or absence of an IGF-II marker and the IGF-I marker on the cells in the test
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`samples, and analyzing the cell population detected by the IGF-I marker and IGF-II marker to
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`identify the tumorcells in the test samples before treatment with the drug candidate compound
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`comparedto after treatment with the drug candidate compound, wherein the presence of a
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`decreased numberof the tumorcells in the specimen after treatment compared to a numberof the
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`tumor cells in a specimen before treatment indicating effectiveness of the drug candidate
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`compoundin treating the cancer in the mammalian subject. The cancer can be metastatic cancer
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`or early stage cancer.
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`BRIEF DESCRIPTION OF THE DRAWINGS
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`[0032]
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`Figure 1 shows human monoclonal antibodies selected against IGF-I that bind to
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`IGF-I or IGF-I and IGF-II.
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`[0033]
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`Figure 2 shows an ELISAbinding assay of IgG 708.2 binding to IGF-I and IGF-
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`Il.
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`cells.
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`[0034]
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`Figure 3 showsthat IgG 708.2 inhibits phosphorylation of IGF-IR in MCF-7
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`[0035]
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`Figure 4 showsthe inhibition of IGF-I binding to soluble IGF-IR by human
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`monoclonal antibodies selected against IGF-I.
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`[0036]
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`Figure 5 shows a dose-dependent inhibition of IGF-II and IGF-I-induced IGF-IR
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`phosphorylation in MCF7cells by anti-IGF-II human antibody IgG1 m708.2.
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`[0037]
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`Figure 6 showsinhibition ofcell motility by IgG1 708.2.
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`[0038]
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`Figure 7 showsthe binding specificity of monoclonal antibodies m705, m706,
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`and m708 to IGF-I and IGF-II by ELISAassay.
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`[0039]
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`Figure 8 shows binding competition of monoclonal antibodies m705, m706 and
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`m708 to binding between IGF-I and IGF-1 receptor.
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`[0040]
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`Figure 9 shows binding of monoclonal antibody m708.2 IgG to IGF-II.
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`DETAILED DESCRIPTION
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`PCT/US2007/066180
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`[0041] The present invention is generally related to antibody compositions and methods
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`for treatment of neoplastic disease in a mammalian subject. The present invention furtherrelates
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`to methods of diagnosing neoplastic disease in a mammalian subject. The antibody compositions
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`are isolated monoclonal antibodies that bind to insulin-like growth factor I. Another set of
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`antibody compositions are monoclonal antibodics that bind to insulin-like growth factor I and arc
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`cross-reactive to and bind to insulin-like growth factor Il. The isolated monoclonal antibodies
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`are, for example, human, non-human primate, rabbit, rat or mouse antibodies. The isolated
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`human monoclonal antibody compositions that bind to insulin-like growth factor I are, for
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`example, m705 and m706. The isolated human monoclonal antibody compositions that bind to
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`both insulin-like growth factor I and insulin-like growth factor II are m708 and m708.2.
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`Monoclonal antibodies m705, m706, m708, and m708.2 do not bind to humaninsulin.
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`[0042]
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`The insulin-like growth factors (IGF) are mitogens that play a role in regulating
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`cell proliferation, differentiation, and apoptosis. The effects of IGFs are mediated through the
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`insulin-like growth factor receptor, IGF-1R. Insulin-like growth factor | (IGF-I) and insulin-like
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`growth factor IT (IGF-II) mediate an effect through binding to type I insulin-like growth factor
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`receptor (IGF-1R). IGF-1R is overexpressed by many tumors and mediates proliferation,
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`motility and protection from apoptosis. Its major ligand which is overexpressed by tumorsis
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`IGF-I. Inhibition of the IGF-IR-mediated signaling can occurat extracellular or intracellular
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`targets. Extracellular IGFs bind to the IGF-IR and the activated tyrosine kinase leads to
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`en