JP2008518902A - Combination therapy of CTLA4 antibody and aromatase inhibitor for breast cancer - Google Patents

Abstract

The present invention relates to anti-CTLA4 antibodies, in particular antibodies 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1. .1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 12.9.1.1, and 10D1 (MDX-010) For the treatment of breast cancer in the form of a human antibody against human CTLA4 in combination with an aromatase inhibitor. More particularly, the present invention relates to the administration of anti-CTLA4 antibodies and exemestane for the treatment of breast cancer.

Description

  Each year, more than 180,000 women are diagnosed with breast cancer in the United States. If current breast cancer rates do not change, each woman born today has a median age of onset of approximately 60-65 years and one in 10 prospects for breast cancer.

  About two-thirds of postmenopausal breast cancer patients have estrogen-dependent disease and are therefore susceptible to anti-estrogen therapy. Estrogens promote the growth and proliferation of specific target cells such as breast epithelial cells and estrogen-dependent breast cancer cells (Brugegmeier Amer J Therapeutics 8: 333-344 (2001)). Therefore, systemic adjuvant therapy administered in addition to the primary treatment of localized breast cancer (eg, breast-conserving surgery and radiation, or mastectomy) is an agent that blocks estrogen receptors on tumor cells. Focus is on. Such antihormonal agents include tamoxifen (NOVALDEX D, SOLTAMOX, tamofen), which inhibits the binding of estrogen to the estrogen receptor, fulvestrant (FASLODEX), which degrades the receptor, and an aromatase enzyme that converts androgen to estrogen. Aromatase inhibitors (eg, anastrozole (ARIMIDEX), letrozole (FEMARA), and exemestane (ALOMASIN)) that block the catalysis of, thus reducing the amount of hormone ligands available for receptor binding included.

  Despite advances in hormone therapy or other breast cancer therapies, it is estimated that 40921 women will die of recurrent or advanced breast cancer in 2004 (American Cancer Society, Cancer Statistics (2004)). Currently there is no curative treatment available for patients with a wide range of diseases, and the therapy is essentially palliative. Therefore, novel therapies such as combining antihormonal agents and immunotherapy are important in breast cancer treatment. More specifically, certain cancer immunotherapy approaches target cytotoxic T lymphocyte binding antigen 4 (CTLA4, CD152), a cell surface receptor expressed on activated T cells. When CTLA4 binds to its natural ligands B7.1 (CD80) and B7.2 (CD86), a negative regulatory signal is delivered to the T cell, and blocking this negative signal results in T cell immune function in animal models. And antitumor activity is enhanced (Thompson and Allison Immunity 7: 445-450 (1997), McCoy and LeGros Immunol. & Cell Biol. 77: 1-10 (1999)). Several studies have demonstrated that blocking CTLA4 using antibodies can significantly enhance T cell-mediated tumor killing and induce anti-tumor immunity (Leach et al., Science 271: 1734-1736 (1996); Kwon et al., Proc. Natl. Acad. Sci. USA 94: 8099-8103 (1997); Kwon et al., Natl. Acad. Sci. USA 96: 15074-15079 (1999)).

  Inducing anti-tumor responses using anti-CTLA4 antibodies is promising in cancer treatment and, despite the promise of anti-hormonal therapy in breast cancer treatment, a novel therapy for breast cancer treatment Is required to develop. The present invention meets this need.

  The present invention includes a method of treating breast cancer in a patient in need thereof. The method includes administering to the patient a therapeutically effective amount of an anti-CTLA4 antibody or antigen-binding portion thereof in combination with a therapeutically effective amount of an aromatase inhibitor.

  In one aspect, the aromatase inhibitor is at least one inhibitor selected from the group consisting of anastrozole, letrozole, and exemestane. In another aspect, the aromatase inhibitor is exemestane.

  In one aspect, the therapeutically effective amount of the human anti-CTLA4 antibody amount ranges from about 1 mg / kg to 40 mg / kg.

  In another aspect, the therapeutically effective amount of the human anti-CTLA4 antibody amount ranges from about 3 mg / kg to 15 mg / kg.

  In yet another aspect, the therapeutically effective amount of exemestane ranges from about 25 mg per day to 200 mg per day.

  In another aspect, the therapeutically effective amount of exemestane is about 25 mg per day.

  In one aspect, the treatment is selected from the group consisting of neoadjuvant chemotherapy, adjuvant therapy, first line therapy, second line therapy, and third line therapy.

  In another aspect, the antibody is selected from the group consisting of a non-human mammalian antibody, a chimeric antibody, and a human antibody.

  In one aspect, the antibody is a human anti-CTLA4 antibody.

In another aspect, the anti-CTLA4 antibody or antigen-binding portion thereof is at least one antibody selected from the group consisting of:
(A) a human antibody having a binding affinity for CTLA4 of about 10 ⁻⁸ or more and inhibiting binding between CTLA4 and B7-1 and binding between CTLA4 and B7-2;
(B) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 116.1 11.7.1. A human antibody having an amino acid sequence comprising at least one human CDR sequence corresponding to a CDR sequence of an antibody selected from the group consisting of: 12.3.1.1, 12.9.1.1, and 10D1;
(C) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 116.1 11.7.1. A human antibody having the heavy and light chain amino acid sequences of an antibody selected from the group consisting of: 12.3.1.1, and 12.9.1.1
(D) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 116.1 11.7.1. A human antibody having an amino acid sequence of a heavy chain variable region and a light chain variable region of an antibody selected from the group consisting of: 12.3.1.1, 12.9.1.1, and 10D1;
(E) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 116.1 11.7.1. , 12.3.1.1, 12.9.1.1, and 10D1 compete for binding of CTLA4 with at least one antibody having an antibody heavy and light chain amino acid sequence selected from the group consisting of An antibody or antigen-binding portion thereof, and (f) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2 .1, 11.6.1, 11.7.1. , 12.3.1.1, 12.9.1.1, and 10D1 cross-competing for the binding of CTLA4 with at least one antibody having heavy and light chain amino acid sequences of an antibody selected from the group consisting of Antibody or antigen-binding portion thereof.

  In another aspect, the antibody is a human antibody having the heavy and light chain amino acid sequences of antibody 11.2.1.

In yet another aspect, the antibody comprises heavy and light chains wherein the amino acid sequences of the heavy chain variable region of the heavy chain and the light chain variable region of the light chain are selected from the group consisting of:
(A) the amino acid sequence of SEQ ID NO: 3 and the amino acid sequence of SEQ ID NO: 9,
(B) the amino acid sequence of SEQ ID NO: 15 and the amino acid sequence of SEQ ID NO: 21,
(C) the amino acid sequence of SEQ ID NO: 27 and the amino acid sequence of SEQ ID NO: 33,
(D) the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 1 and the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 7,
(E) the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 13 and the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 19,
(F) the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 25 and the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 31;
(G) Amino acid sequence of the heavy chain variable region and light chain variable region of antibody 10D1.

In one aspect, the antibody or antigen-binding portion thereof is an antibody selected from the group consisting of:
(A) having a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, and further comprising the amino acid sequences set forth in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 An antibody having a light chain variable region,
(B) having a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, and further comprising the amino acid sequences set forth in SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24 An antibody having a light chain variable region,
(C) having a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, and further comprising the amino acid sequences set forth in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36 An antibody having a light chain variable region; and (d) a heavy chain variable region comprising the heavy chain CDR1, CDR2, and CDR3 amino acid sequences of antibody 10D1, and further the light chain CDR1, CDR2, and CDR3 amino acids of antibody 10D1 An antibody having a light chain variable region comprising a sequence.

  In another aspect, the method comprises an alkylating agent, a folic acid antagonist, a pyrimidine antagonist, an anthracycline antibiotic, a platinum compound, a taxane, a vinca alkaloid, a camptothecin analog, a toll-like receptor agonist, a heat shock protein. Tumor vaccine as main agent, therapy based on antigen presenting cells, mammalian rapamycin target protein inhibitor, erbB2 inhibitor, EGFR inhibitor, VEGF inhibitor, VEGFR inhibitor, angiogenesis inhibitor, antibody, immunomodulator, selective estrogen It further comprises administering to the patient at least one agent selected from the group consisting of receptor modulators, cytokines, tumor vaccines, antiproliferative drugs, immune co-stimulatory molecules, and cytokines.

  The present invention includes a pharmaceutical composition for the treatment of breast cancer comprising a therapeutically effective amount of an anti-CTLA4 antibody, a therapeutically effective amount of an aromatase inhibitor, and a pharmaceutically acceptable carrier.

  In one aspect, the aromatase inhibitor is at least one aromatase inhibitor selected from the group consisting of anastrozole, letrozole, and exemestane.

  In another aspect, the aromatase inhibitor is exemestane.

  The invention includes the use of an amount of an anti-CTLA4 antibody in preparing a composition for the treatment of breast cancer in a patient, wherein the treatment further comprises administering an amount of an aromatase inhibitor to the patient.

  In one aspect, the aromatase inhibitor is exemestane and the antibody is a human antibody having the heavy chain and light chain variable region amino acid sequences of antibody 11.2.1.

  The foregoing summary, as well as the following detailed description of the invention, will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, there are shown in the drawings embodiments which are presently preferred. However, it should be understood that the invention is not limited to the detailed schemes and instrumentalities shown.

  The present invention relates to anti-CTLA4 antibodies used in combination with at least one aromatase inhibitor such as anastrozole, letrozole, and exemestane to treat breast cancer in patients in need of such treatment. . The invention further relates to the treatment of breast cancer with an antibody-aromatase inhibitor combination in combination with one or more other agents.

  Unless otherwise defined herein, scientific and technical terms used in connection with the present invention shall have the meanings that are commonly understood by those of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. In general, the nomenclature used in connection with and in the cell and tissue culture, molecular biology, immunology, microbiology, genetics, protein and nucleic acid chemistry, and hybridization methods described herein. Are well known and commonly used in the industry.

  The methods and techniques of the present invention generally follow various methods well known in the art, unless otherwise specified, and various general and more detailed references cited and discussed throughout this specification. As described in. Such references include, for example, Sambrook and Russell, “Molecular Cloning, A Laboratory Approach”, Cold Spring Harbor Press, Cold Spring Harbor, New York (2001), Ausubel et al., “Current Protocol,” John Wiley & Sons, New York, USA (2002), and Harlow and Lane, “Antibodies: A Laboratory Manual”, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1990) Are hereby incorporated by reference. Enzymatic reactions and purification techniques are performed according to manufacturer's specifications, generally as is done in the art or as described herein. The nomenclature used in connection with the analytical chemistry, synthetic organic chemistry, and pharmaceutical and pharmaceutical chemistry described herein and in these experimental procedures and techniques is well known and commonly used in the art. Is. Standard techniques are used for chemical synthesis, chemical analysis, pharmaceutical formulation, formulation form, delivery, and patient treatment.

  As used herein, each of the following terms has the meaning associated with it in this section.

  As used herein, the articles “a” and “an” are used to refer to one or more (ie, at least one) of the grammatical objects of the article. By way of example, “an element” means one element or a plurality of elements.

  As used herein, the twenty conventional amino acids and their abbreviations follow conventional usage. See "Immunology--A Synthesis" (2nd edition, edited by ES Golub and DR Glenn, Sinauer Associates, Sunderland, Mass., USA), incorporated herein by reference. .

  A “conservative amino acid substitution” is one in which the amino acid residue is replaced with another amino acid residue having a side chain R group with similar chemical properties (eg, charge or hydrophobicity). In general, conservative amino acid substitutions do not substantially change the nature of the functional group of the protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent degree of sequence identity or similarity can be adjusted upwards to correct the conservative nature of the substitution. Means for making this adjustment are well known to those skilled in the art. For example, Pearson, Methods Mol. Biol. 243: 307-31 (1994).

  Examples of amino acid groups having side chains with similar chemical properties include 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine, 2) aliphatic-hydroxyl side chains: serine and threonine, 3) Amide-containing side chains: asparagine and glutamine, 4) aromatic side chains s: phenylalanine, tyrosine, and tryptophan, 5) basic side chains: lysine, arginine, and histidine, 6) acidic side chains: aspartic acid and glutamic acid, and 7) Sulfur-containing side chain: includes cysteine and methionine. Preferred conservative amino acid substituents are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine.

  Alternatively, a conservative substitution refers to a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al., Science 256: 1443-45 (1992), which is incorporated herein by reference. Any change you have. A “moderately conservative” replacement is any change having a non-negative value in the PAM250 log-likelihood matrix.

  Preferred amino acid substitutions (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinity to form protein complexes, (4) such It imparts or modifies other physicochemical or functional properties of the analog. Analogs containing substitutions, deletions, and / or insertions may include various mutant proteins of sequences other than the specified peptide sequence. For example, one or more amino acid substitutions (preferably conservative amino acid substitutions) are made in a specified sequence (preferably outside the domain that forms the intermolecular contact, eg, the polypeptide portion outside the CDRs). Things can be used. Conservative amino acid substitutions should not substantially change the structural properties of the parent sequence (e.g., substituted amino acids destroy a helix present in the parent sequence or other types of secondary characterizing the parent sequence). Should not tend to disrupt the structure). Examples of secondary and tertiary structures of polypeptides recognized in the art are “Proteins, Structures and Molecular Principles” (Creighton ed., WH Freeman and Company, New York, USA (1984)), “Introduction to Protein Structure "(edited by C. Branden and J. Tokyo, edited by Garland Publishing, New York, NY (1991)), and Thornton et al., Nature 354: 105 (1991). Each of which is incorporated herein by reference.

  Polypeptide sequence similarity is usually measured using sequence analysis software. Protein analysis software matches similar sequences using similarity measures assigned to various substitutions, deletions, and other changes, including conservative amino acid substitutions. For example, the Genetics Computer Group (GCG, available from Genetics Computer Group, Inc.), also called Wisconsin Package, is an integrated suite of over 130 programs for accessing, analyzing, and manipulating nucleotide and protein sequences. Software package. GCG includes programs such as “Gap” and “Best Fit”, using default parameters, between closely related polypeptides, such as homologous polypeptides from different species, or with wild-type proteins Sequence similarity, homology, and / or sequence identity between the mutant proteins can be determined. See, for example, GCG version 6.1, version 9.1, and version 10.0.

  Polypeptide sequences can be compared using FASTA using default or recommended parameters, a program of GCG version 6.1. FASTA (eg, FASTA2 and FASTA3) provides the best partially overlapping region alignment and percent sequence identity between query and search sequences (Pearson, Methods Enzymol. 183: 63-98 (1990). Pearson, Methods MoI. Biol., 132: 185-219 (2000)). Another preferred algorithm when comparing the sequences of the present invention to a database containing a large number of sequences from different organisms is the computer program BLAST, in particular blastp or tblastn, using default parameters. See, for example, Altschul et al., J. MoI. Mol. Biol. 215: 403-410 (1990); Altschul et al., Nucleic Acids Res. 25: 3389-402 (1997).

“Aromatase” as used herein refers to an enzyme (eg, 450 _arom ) that catalyzes the aromatization of the A ring of an androgen substrate (mainly androstenedione) to estrone.

  An “aromatase inhibitor” is a compound that, when contacted with aromatase, inhibits the formation of the A ring of an androgen substrate to a detectable extent compared to the level of aromatization in the absence of the compound or substance or Means a substance. Any assay that assesses the level of enzyme activity by aromatase can be used to assess whether there is and / or level of inhibition. Aromatase inhibitors include, but are not limited to, steroidal and nonsteroidal inhibitors such as exemestane, anastrozole, letrozole, fadrozole, borozole, and formestane, which are currently known Including any aromatase inhibitor, whether it has been identified or identified in the future.

An intact “antibody” comprises at least two heavy (H) chains and two light (L) chains linked together by disulfide bonds. See generally, “Fundamental Immunology”, Chapter 7 (Paul, W. Ed., 2nd edition, Raven Press, New York, USA (1989)), which is incorporated by reference in its entirety. Each heavy chain is composed of a heavy chain variable region (HCVR or V _H ) and a heavy chain constant region (C _H ). The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (LCVR or V _L ) and a light chain constant region. The light chain constant region is comprised of one domain, C _L. The _VH and _VL regions are further subdivided into hypervariable regions termed complementarity determining regions (CDRs) interspersed with more conserved regions termed framework regions (FR). Can do. Each V _H and V _L is composed of three CDRs and four FRs arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxyl terminus. The assignment of amino acids to each domain can be found in Kabat, “Sequences of Proteins of Immunological Interest” (National Institutes of Health, Bethesda, Maryland, USA (1987 and 1991)), or Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987); Chothia et al., Nature 342: 878-883 (1989).

The term “antigen-binding portion” of an antibody (or simply “antibody portion”) as used herein refers to one or more antibody fragments that retain the ability to specifically bind to an antigen (eg, CTLA4). . It has been shown that the antigen binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments included within the term “antigen-binding portion” of an antibody include (i) a Fab fragment consisting of the monovalent fragments V _L , V _H , C _L , and C _H 1 domains, (ii) ) F (ab ′) ₂ fragment, ie a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge, (iii) an Fd fragment consisting of V _H and C _H 1 domains, (iv) an antibody Fv fragment consisting of the V _L and V _H domains, (v) the dAb fragment consisting of the V _H domain (Ward et al., (1989) Nature 341: 544-546), and (vi ) An isolated complementarity determining region (CDR) is included. In addition, the two domains of the Fv fragment, _VL and _VH , are encoded by separate genes, but a pair of _VL and _VH regions are known as (single chain Fv (scFv)) ) Recombinant methods can be used to join together with a synthetic linker that results in a single protein chain that forms a monovalent molecule. See, for example, Bird et al., Science 242: 423-426 (1988) and Huston et al., Proc. Natl. Acad. Sci. USA 85: 5879-5883 (1988)). Such single chain antibodies are also intended to be encompassed within the term “antigen-binding portion” of an antibody. Other forms of single chain antibodies such as diabodies are also included. Diabodies are bivalent bispecific antibodies in which the V _H and V _L domains are expressed on a single polypeptide chain, but are too short to allow pairing between two domains on the same chain. A disabling linker is used, thereby allowing the domain to pair with the complementary domain of another chain, giving two antigen binding sites (eg, Holliger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993); see Poljak et al., Structure 2: 1121-1123 (1994)).

Furthermore, the antibody or antigen binding portion thereof may be a portion of a larger immunoadhesion molecule produced by covalent or non-covalent binding of the antibody or antibody portion and one or more other proteins or peptides. Examples of such immunoadhesion molecules are those that use the streptavidin core region to generate tetrameric scFv molecules (Kipriyanov et al., Human Antibodies and Hybridomas 6: 93-101 (1995)). And a bivalent biotin-labeled scFv molecule using a cysteine residue, a marker peptide, and a C-terminal polyhistidine tag (Kipriyanov et al., Mol. Immunol. 31: 1047-1058 (1994)). Year)). Another example is an immunoadhesin in which one or more CDRs from an antibody are covalently or non-covalently incorporated into a molecule to specifically bind that molecule to an antigen of interest, such as CTLA4. An example is included. In such embodiments, the CDR may be incorporated as part of a larger polypeptide chain, may be covalently linked to another polypeptide chain, or may be incorporated non-covalently. Antibody portions such as Fab and F (ab ′) ₂ fragments can be prepared from whole antibodies using conventional techniques such as papain or pepsin digestion of whole antibodies, respectively. Further, as described herein, standard recombinant DNA techniques can be used to obtain antibodies, antibody portions, and immunoadhesion molecules.

It should be understood that where an “antibody” is referred to herein with respect to the present invention, its antigen binding portion may be used. The antigen binding portion competes with the intact antibody for specific binding. See generally, “Fundamental Immunology”, Chapter 7 (Paul, W. Ed., 2nd Edition, Raven Press, New York, NY, 1989) (however, incorporated by reference in its entirety). Antigen-binding portions can be generated by recombinant DNA techniques or by enzymatic or chemical resolution of intact antibodies. In some embodiments, the antigen binding portion includes fragments such as Fab, Fab ′, F (ab ′) ₂ , Fd, Fv, dAb, and complementarity determining region (CDR), single chain antibodies (scFv), Chimeric antibodies, diabodies, as well as polypeptides comprising at least a portion of an antibody sufficient to bind a particular antigen to the polypeptide are included. In embodiments having one or more binding sites, the binding sites may be the same or different from one another.

  The term “human antibody” or “human sequence antibody” as used interchangeably herein includes antibodies having a variable region and a constant region (if present) derived from human germline immunoglobulin sequences. Human sequence antibodies of the invention are amino acid residues that are not encoded by human germline immunoglobulin sequences (eg, mutations introduced in vitro by random or site-specific mutagenicity or in vivo by somatic mutation). May be included. However, the term “human antibody” as used herein refers to a “chimeric” antibody (ie, “humanized”) in which a CDR sequence derived from the germline of another mammalian species, such as a mouse, is joined to a human framework sequence. "Or PRIMATIZED ™ antibody).

  The term “chimeric antibody” as used herein means an antibody that comprises regions from two or more different antibodies. In one embodiment, one or more of the CDRs are from a human anti-CTLA4 antibody. In another embodiment, all of the CDRs are derived from a human anti-CTLA4 antibody. In another embodiment, CDRs from multiple human anti-CTLA4 antibodies are combined into a chimeric human antibody. For example, the chimeric antibody may comprise CDR1 from a first human anti-CD40 antibody light chain, CDR2 from a second human anti-CTLA4 antibody light chain, and CDR3 from a third human anti-CTLA4 antibody light chain; The CDRs from the heavy chain may be derived from one or more other anti-CD40 antibodies. Further, the framework region may be obtained from one of the same anti-CTLA4 antibodies or from one or more different humans.

  In addition, as discussed previously herein, chimeric antibodies include antibodies that contain portions from multiple species germline sequences.

  “Glycoform” refers to a complex oligosaccharide structure comprising a chain of various carbohydrate units. Such a structure is described, for example, in “Essentials of Glycobiology”, edited by Varki et al., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1999), which is a standard glycobiology. An overview of nomenclature is also provided. Such glycoforms include, but are not limited to, G2, G1, G0, G-1, and G-2 (see, eg, International Patent Publication WO 99/22764).

  A “glycosylation pattern” is a covalent bond to a protein (eg, a glycoform), as well as a site where the glycoform is covalently attached to the peptide backbone of the protein, and more particularly to an immunoglobulin protein. Is defined as the pattern of carbohydrate units bound together.

  Antibodies expressed by different cell lines or in transgenic animals probably have different glycoforms and / or glycosylation patterns compared to each other. However, regardless of the glycosylation of the antibody, all antibodies encoded by the nucleic acid molecules provided herein or comprising the amino acid sequences provided herein are part of the invention.

  The term “effective amount” or “therapeutically effective amount” as used herein mediates a detectable therapeutic response when administered to a mammal, preferably a human, as compared to the response detected in the absence of the compound. Means quantity. Without limitation, therapeutic response such as suppression and / or reduction of tumor growth, tumor size, metastasis, etc. is readily assessed by any method recognized in the art including, for example, the methods disclosed herein. can do.

  The effective amount of the compound or composition administered herein will vary and will vary depending on the disease or condition being treated, the stage of the disease, the age, health condition, physical condition, severity of the disease being treated. One of ordinary skill in the art will appreciate that it can be readily determined based on several factors such as the particular compound being administered.

  The term “competing” as used herein means that for an antibody, the first antibody or antigen-binding portion thereof competes with the second antibody or antigen-binding portion thereof for binding, wherein The binding of one antibody to its cognate epitope is detectably reduced in the presence of the second antibody compared to the binding of the first antibody in the absence of the second antibody. There may be an option where the binding of the second antibody to its epitope is detectably reduced in the presence of the first antibody, but this need not be the case. That is, the first antibody can inhibit the binding of the second antibody to its epitope, and the second antibody does not inhibit the binding of the first antibody to its respective epitope. However, if each antibody detectably inhibits the binding of another antibody to its cognate epitope or ligand to the same, greater, or lesser extent, the antibodies will bind each other for binding of their respective epitopes. Say “cross-compete”. For example, a cross-competing antibody can bind to an epitope or portion of an epitope that is a binding partner of an antibody used in the present invention. The use of both competing and cross-competing antibodies is encompassed by the present invention. Regardless of the mechanism by which such competition or cross-competition occurs (eg, steric hindrance, conformational change, or binding to a common epitope or portion thereof, etc.), one of ordinary skill in the art will be provided herein. Based on the teachings, it should be appreciated that such competing antibodies and / or cross-competing antibodies are included and useful for the methods disclosed herein.

  The term “epitope” includes any protein determinant capable of specific binding to an immunoglobulin or T cell receptor. Epitopic determinants usually consist of chemically active surface molecule groups such as amino acids or sugar side chains and usually have specific three-dimensional structural characteristics as well as specific charge characteristics. The conformation and non- conformation epitopes are distinguished in that the binding to the former but not the latter is impaired in the presence of a denaturing solvent.

  “Explanatory material”, as the term is used herein, refers to the present invention in a kit for affecting, alleviating or treating the various diseases or disorders listed herein. Includes publications, records, diagrams, or any other representation medium that can be used to communicate the usefulness of the compounds, combinations, and / or compositions. Optionally or alternatively, the explanatory material may also describe one or more methods for alleviating a cell, tissue, or mammalian disease or disorder, including those disclosed elsewhere herein. it can.

  The instructional material for the kit may be attached to, for example, a container containing the compound and / or composition of the present invention, or shipped with a container containing the compound and / or composition. Alternatively, the instructional material may be shipped separately from the container with the intention that the recipient uses the instructional material and the compound cooperatively.

  Except as otherwise noted, the terms “patient” or “subject” are used interchangeably, mammals such as human patients and non-human primates, and veterinary subjects such as rabbits, rats, mice, and other Refers to animals. The patient preferably refers to a human.

  Herein, conventional notation is used to delineate polypeptide sequences, ie, the left-hand side of polypeptide sequences is the amino terminus and the right-hand side of polypeptide sequences is the carboxyl terminus.

The expression “specifically binds” as used herein is a compound that recognizes and binds to a particular molecule but does not substantially recognize or bind to other molecules in the sample, such as a protein, nucleic acid, antibody, etc. Means. For example, an antibody or peptide inhibitor recognizes and binds a cognate ligand in a sample (eg, an anti-CTLA4 antibody that binds to its cognate antigen CTLA4) but does not substantially recognize or bind other molecules in the sample. . Thus, under the indicated assay conditions, a specified binding moiety (eg, an antibody or antigen binding portion thereof) binds preferentially to a particular target molecule and is significant to other components present in the test sample. Do not combine by quantity. A variety of assay formats can be used to select antibodies that specifically bind the molecule of interest. For example, solid phase ELISA immunoassay, immunoprecipitation, BIAcore, and Western blot analysis are used to identify antibodies that specifically react with CTLA4. Typically, a specific or selective reaction will be at least twice background signal or noise, more typically more than 10 times background, and even more specifically, the equilibrium dissociation constant (K _D ). Is ≦ 1 μM, preferably ≦ 100 nM, most preferably ≦ 10 nM, an antibody is said to “specifically bind” an antigen.

The term “K _D ” refers to the equilibrium dissociation constant of a particular antibody-antigen interaction.

  As used herein, “substantially pure” is the main species in which the species of interest is present (ie, more abundant than any other individual species in the composition on a molar basis). Preferably, the substantially purified fraction comprises at least about 50 percent (in molarity) of all macromolecular species in which the target species (eg, anti-CTLA4 antibody) is present. It is preferable that In general, a substantially pure composition will comprise more than about 80 percent, more preferably more than about 85%, 90%, 95%, and 99% of all macromolecular species present in the composition. The target species is purified and essentially homogeneous (contaminant species cannot be detected in the composition by conventional detection methods) and the composition is essentially a single macromolecular species Most preferably it consists of:

  As used herein, “treating” refers to the frequency with which a patient experiences symptoms of a disease (ie, tumor growth and / or metastasis, or other effects mediated by immune cell number and / or activity). Means to decrease. The term is used to administer a compound or agent of the invention to prevent or delay the occurrence of symptoms, complications, or biochemical signs of disease (eg, elevated levels of PSA), alleviate symptoms, or disease, Including preventing or suppressing further development of the condition or disorder. Treatment may be prophylactic (to prevent or delay the onset of the disease, or prevent its clinical or subclinical symptoms from appearing), or the therapeutic suppression or reduction of symptoms after the onset of the disease. But you can.

I. TECHNICAL FIELD The present invention relates to a method for treating breast cancer by administering human CTLA4 or an antibody that binds an antigen-binding portion of this antibody in combination with hormone (eg, aromatase inhibitor) therapy. The combination of blocking CTLA4 and reducing patient estrogen levels according to the methods of the present invention provides a synergistic therapeutic benefit, as discussed more fully below.

  While the present invention includes numerous combination therapies in which an antibody is administered to a patient in combination with at least one aromatase inhibitor, the present invention is in no way limited to exemplary agents that are merely for the purpose of illustration described herein. .

  In one embodiment, a combination of a CTLA4 antibody or antigen-binding portion thereof and an aromatase inhibitor (“AI”) is administered to a breast cancer patient. In one aspect, the aromatase inhibitor is selected from the group consisting of anastrozole, letrozole, and exemestane, commercially available as ARIMIDEX, FEMARA, and AROMASIN, respectively. More preferably, the aromatase inhibitor is exemestane. This is because the synergistic or additive effect is via administration of a combination comprising anti-CTLA4 antibody and AI. Without being bound to a particular theory of the present invention, AI therapy may increase the immune response to tumor cells, as it may increase exposure of the breast cancer tumor-specific antigen to the immune system, possibly through changes in apoptosis or other architecture. To do. That is, hormonal therapy can create or increase the source of tumor-specific antigens that mediate tumor cell death, thereby supplying tumor antigens to the host's antigen presentation pathway. Anti-CTLA4 antibodies mediate an increased immune response to increased levels of tumor-specific antigens in the antigen presentation pathway, and therefore provide a potential synergistic therapeutic effect when combined with aromatase inhibitor therapy. Other combination therapies that provide a synergistic effect of anti-CTLA4 enhancement of immune responses through the release of tumor-specific antigen cell death are radiation, surgery, and chemotherapy, among others.

  Without being bound to a particular theory of the present invention, the combination of CTLA4 block and aromatase inhibition may elicit a more robust immunological response against breast cancer. Thus, in particular, the combination of an aromatase block with exemestane with an anti-CLTA4 antibody can provide a potential synergistic effect, thereby providing an important new therapeutic treatment for breast cancer.

  The method of the present invention is a neoadjuvant prior to surgery, radiation therapy, or any other therapeutic localized treatment to sensitize tumor cells or otherwise confer therapeutic benefit to the patient. Can be performed as a therapy. However, the present invention is not limited to the setting of neoadjuvant. Rather, the methods of the present invention can be used along with the overall disease and treatment flow, such as, but not limited to, adjuvants for breast cancer, first line, second line, third line therapy, and the like.

  In one embodiment, breast cancer is estrogen receptor positive (ER +), so aromatase inhibition reduces estrogen levels and thereby affects the tumor. In one aspect, since ER + tumors are present in post-men women women, reducing estrogen levels mediates therapeutic anti-tumor effects in such patients. However, although it may be preferable to administer AI therapy to postmenopausal ER + patients, the present invention is not limited to such patients, but rather is determined by one of ordinary skill in the art with the teachings presented herein. Administration of anti-CTLA4 antibody-aromatase inhibitor therapy to any patient who can benefit from the combination of reduced estrogen levels and immune enhancement with CTLA4 block.

II. Dosage regimen The dosage regimen may be adjusted to provide the optimum desired response. For example, a single bolus administration can be performed, the dosage can be divided into several times, administered over time, or the dosage can be increased or decreased depending on the urgency of the treatment situation. It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, refers to a physically discrete unit suitable as a unit dosage for a mammalian subject undergoing treatment, each unit being combined with the required pharmaceutical carrier for the desired treatment. Contains a predetermined amount of active compound calculated to produce an effect. Details of the dosage unit forms of the present invention include: (a) the unique characteristics of the antibody as well as the specific therapeutic or prophylactic effect to be achieved; (b) such active compounds tailored to the individual therapeutic sensitivity. It is inevitably determined by the industry-specific restrictions of blending and directly depends on these.

  Exemplary non-limiting ranges for therapeutically effective amounts of antibody administered are, according to the present invention, at least about 1 mg / kg, at least about 5 mg / kg, at least about 10 mg / kg, greater than about 10 mg / kg, or At least about 15 mg / kg, such as about 1-30 mg / kg, or such as about 1-25 mg / kg, or such as about 1-20 mg / kg, or such as about 5-20 mg / kg, or such as about 10-20 mg / kg, Or for example about 15-20 mg / kg, or for example about 15 mg / kg. It should be noted that dosage values may vary depending on the type and severity of the condition to be alleviated and may include one or more doses. Any particular subject, detailed dosage regimen should be adjusted over time according to individual requirements and the professional judgment of the person performing or supervising the administration of the composition, as described herein. It is further to be understood that the dosage ranges are exemplary only and do not limit the scope or operation of the claimed compositions. It should also be understood that the determination of appropriate dosages and dosing schedules for antibodies is well known in the relevant art and can be accomplished by one of ordinary skill in the art once the teachings disclosed herein are presented. .

  In one embodiment, the antibody is administered as an intravenous formulation as a sterile aqueous solution containing about 5-20 mg / ml antibody in a suitable buffer system.

  In one embodiment, for low dose administration, a portion of the dose is administered by intravenous bolus administration and the remainder is administered by infusion of the antibody formulation. For example, an intravenous injection of 0.01 mg / kg of antibody can be given as a bolus dose and the remainder of a given antibody dose can be administered by intravenous injection. In another embodiment, the entire low dose is administered as a single bolus dose. For higher doses, eg 3 mg / kg, the antibody is not administered as a bolus dose, but the whole dose is administered by infusion. A given dose of antibody can be administered, for example, over a period of about 1.5 hours to about 5 hours.

  The present invention relates to the administration of a combination of an anti-CTLA4 antibody and an aromatase inhibitor. One skilled in the art will know that the combination may be administered at the same time or the two agents may be administered at different times. For example, in one embodiment, the antibody is administered as a single dose and / or infusion, and the aromatase inhibitor (eg, exemate) is initiated before, during, or after antibody administration. Administer once a day. Normally, exemestane is administered daily, usually at the same time of day and once daily after meals. However, the present invention is not limited to any particular dose or dosing schedule for an aromatase inhibitor. Rather, optimal dosages, routes, and dosing schedules for antibody and aromatase inhibitor administration can be readily determined by those skilled in the relevant art using well-known methods.

  For example, a single dose or multiple doses of antibody can be administered. Alternatively, at least one dose, or at least 3, 6, or 12 doses can be administered. This dose may be, for example, every 2 weeks, every month, every 20 days, every 25 days, every 28 days, every 30 days, every 40 days, every 6 weeks, every 50 days, every 2 months, every 70 days, every 80 days, every 3 months, every 6 months It can be administered every or every year. In addition, aromatase inhibitors are daily, several times or once a day, weekly, every other week, every three weeks, every four weeks, every month, every three months, every six months, once a year, or skilled It can be administered at any other time as determined by the practitioner that provides a therapeutic benefit to the patient.

  The antibody can be administered until disease progression or untolerated toxicity, or until 12 consecutive cycles are reached, whichever is shorter. The antibody may be administered using a dosing regimen that includes administration of a lower dose after the initial loading dose. Tumor recurrence experience, where the repetitive process of antibody and aromatase inhibitor consisting of at least one, and more preferably several cycles, has previously benefited from the combination of anti-CTLA4 antibody and aromatase inhibitor Can be administered to certain patients.

  In one embodiment, the patient is administered a single injection comprising anti-CTLA4 antibody intravenously every 28 days at a dose of about 3 mg / kg. A dose of about 25 mg exemestane per day is also administered to the patient before, during, and / or after antibody administration. In an embodiment of the invention, the patient is given a single dose comprising anti-CTLA4 antibody of about 3 mg / kg, preferably about 6 mg / kg, more preferably about 10 mg / kg, preferably about 15 mg / kg for 3 months. Administer intravenously every time. A dose of about 25 mg exemestane per day is also administered to the patient before, during, and / or after antibody administration. In another embodiment, a patient is administered a bolus dose comprising an anti-CTLA4 antibody every 28 days at a dose greater than 3 mg / kg (eg, 6, 10, 15, 20, 25, 30, 35, or 40 mg / kg). Administer intravenously. The dose can be adjusted based on toxicity (if any) and therapeutic efficacy, among other factors, as is known in the art. A dose of about 25 mg exemestane per day is also administered to the patient before, during, and / or after antibody administration.

  The antibody-aromatase inhibitor combination can be administered as a first-line systemic adjuvant therapy, or surgery, radiation therapy, to sensitize tumor cells or otherwise confer therapeutic benefit to the patient, Or it can be administered to the patient as neoadjuvant therapy prior to any other treatment.

  Furthermore, this combination can be administered as a second line therapy, such as, but not limited to, if tamoxifen first line therapy has failed once. Alternatively, the combination can be administered in parallel with tamoxifen therapy and / or at any time during tamoxifen therapy, usually administered for about 5 years after the first treatment. In another embodiment, the combination of anti-CTLA4 antibody and exemestane can be administered to a patient after about 2 or 3 years of tamoxifen or other adjuvant therapy. This is because exemestane therapy after about 2-3 years of tamoxifen therapy has been demonstrated to provide therapeutic benefits while reducing the side effects of tamoxifen therapy.

  Therefore, the combination of an anti-CLTA4 antibody and an aromatase inhibitor, preferably exemestane, is a systemic adjuvant therapy that uses another aromatase inhibitor (eg, anastrozole, letrozole, etc.), particularly if tamoxifen therapy has failed once. Can be beneficial once and / or after about 2-3 years of tamoxifen administration. Thus, as will be appreciated by those of skill in the art based on the disclosure provided herein, the present invention includes additional antihormones, including but not limited to tamoxifen, anastrozole, letrozole, and fulvestrant. Including or subsequent to administration of an anti-CTLA4 antibody and an aromatase inhibitor in combination with therapy.

III. Anti-CTLA4 Antibody In one embodiment, the CTLA4 antibody used in the present invention comprises a heavy chain in which the amino acid sequence of _VH comprises the amino acid sequences set forth in SEQ ID NOs: 3, 15, and 27. In yet another embodiment, the V _L of the CTLA4 antibody comprises the amino acid sequence set forth in SEQ ID NOs: 9, 21, and 33. More preferably, the V _H and V _L regions of the antibody have an amino acid sequence set forth in SEQ ID NO: 3 (V _H 4.1.1) and SEQ ID NO: 9 (V _L 4.1.1), SEQ ID NO: 15 ( V _H 4.13.1) and amino acid sequences set forth in SEQ ID NO: 21 (V _L 4.13.1), and SEQ ID NO: 27 (V _H 11.2.1) and SEQ ID NO: 33 (V _L 11. 2.1) each containing the amino acid sequence described. Most preferably, the antibody is ticilimumab (also known as CP-675,206) having the heavy and light chain amino acid sequences of antibody 11.2.1.

  In yet another embodiment, the heavy chain amino acid sequence comprises an amino acid sequence encoded by a nucleic acid comprising the nucleic acid sequences set forth in SEQ ID NOs: 1, 13, and 25. In yet another embodiment, the light chain comprises an amino acid sequence encoded by a nucleic acid comprising the nucleic acid sequence set forth in SEQ ID NOs: 7, 19, and 31. More preferably, the heavy and light chains are the nucleic acid sequences set forth in SEQ ID NO: 1 (heavy chain 4.1.1) and SEQ ID NO: 7 (light chain 4.1.1), respectively, SEQ ID NO: 13 (heavy chain 4 .13.1) and SEQ ID NO: 19 (light chain 4.13.1), respectively, and SEQ ID NO: 25 (heavy chain 11.2.1) and SEQ ID NO: 31 (light chain 11.2.1). The amino acid sequence encoded by the nucleic acid comprising the nucleic acid sequence described in

In addition, the antibody comprises a human CDR amino acid sequence derived from the V _H 3-30 or 3-33 gene, or a heavy chain amino acid sequence comprising conservative substitutions or somatic mutations therein. It will be appreciated that the V _H 3-30 or V _H 3-33 gene encodes FR1 to FR3 of the heavy chain variable region of the antibody molecule. Therefore, the present invention provides 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11. 2.1, 11.6.1, 11.7.1, 12.3.1.1, 2.9.1.1, 10D1, and FR1 to FR3 of an antibody selected from the group consisting of DP-50 Up to at least 85%, more preferably at least 90%, even more preferably at least 91%, even more preferably at least 94%, even more preferably at least 95%, more preferably at least 97%, Even more preferably it comprises antibodies that are at least 98%, even more preferably at least 99%, most preferably 100%.

  The antibody may further comprise a CDR region in its light chain derived from the A27 or O12 gene, or 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 2.9.1. The CDR region of an antibody selected from the group consisting of 1, 10D1 may be included.

In other embodiments of the invention, the antibody inhibits binding between CTLA4 and B7-1, B7-2, or both. The antibody inhibits binding to B7-1 with an IC ₅₀ of about 100 nM or less, more preferably about 10 nM or less, such as about 5 nM or less, even more preferably about 2 nM or less, or even more preferably about 1 nM or less. Preferably it can be done. Similarly, the antibody has a binding to B7-2 of about 100 nM or less, more preferably 10 nM or less, such as even more preferably about 5 nM or less, even more preferably about 2 nM or less, or even more preferably about 1 nM or less. Can be inhibited with IC ₅₀ .

Furthermore, in another embodiment, the anti-CTLA4 antibody has an affinity for binding to CTLA4 of about 10 ⁻⁸ or higher, more preferably about 10 ⁻⁹ or higher, more preferably about 10 ⁻¹⁰ or higher. And even more preferably an affinity of about ^10-11 or greater.

  The anti-CTLA4 antibody includes heavy and light chains of an antibody selected from the group consisting of 4.1.1, 6.1.1, 11.2.1, 4.13.1, and 4.14.3 Antibodies that compete for binding with an antibody having an amino acid sequence are included. Furthermore, anti-CTLA4 antibodies can compete for binding with antibody 10D1.

  In another embodiment, the antibody comprises antibody 4.1.1, 4.13.1, 4.14.3, 6.1.1, or 11.2.1 heavy and light chain sequences, variable heavy. It is preferred to cross-compete with antibodies having chain and variable light chain sequences and / or heavy and light chain CDR sequences. For example, the antibody is a heavy and light chain amino acid of an antibody selected from the group consisting of 4.1.1, 4.13.1, 4.14.3, 6.1.1, or 11.2.1 It can bind to an epitope that is a binding partner of an antibody having a sequence, variable sequence, and / or CDR sequence. In another embodiment, the antibody cross-competes with an antibody having heavy and light chain sequences or antigen binding sequences of 10D1 (MDX-D010).

  In another embodiment, the present invention provides 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1. CDR-1 of an antibody selected from the group consisting of 1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, and 12.9.1.1, A heavy chain comprising the amino acid sequences of CDR-2 and CDR-3 and a light chain comprising the amino acid sequences of CDR-1, CDR-2 and CDR-3, or conservative changes (of nonpolar residues) Substitution with other non-polar residues, substitution of polar charged residues with other polar uncharged residues, substitution of polar charged residues with other polar charged residues, residues with similar structures Selected from the group consisting of substitutions), non-conservative substitutions (substitutions that become polar charged residues instead of polar uncharged residues), and nonpolar residues instead of polar residues Selected from the group consisting of substitutions), additions, and performed using an anti-CTLA4 antibody comprising a sequence having changes from the CDR sequences selected from the group consisting of deletion.

  In another embodiment of the invention, the antibody comprises fewer than 10, 7, 5, or 3 amino acid changes from the germline sequence in the framework or CDR regions. In another embodiment, the antibody comprises less than 5 amino acid changes in the framework regions and less than 10 changes in the CDR regions. In a preferred embodiment, the antibody comprises fewer than 3 amino acid changes in the framework regions and fewer than 7 changes in the CDR regions. In a preferred embodiment, changes in the framework regions are conservative and changes in the CDR regions are somatic mutations.

  In another embodiment, the antibody is for antibody 3.1.1, 4.1.1, 4.8.1, 4.10. For heavy and light chain full length sequences or for heavy chain or light chain alone. 2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 12.9. 1.1 Sequence (eg, amino acid, nucleic acid, or both) identity or sequence similarity with the sequence of 10D1 is at least 80%, more preferably at least 85%, even more preferably at least 90%, even more preferably Is at least 94%, preferably at least 95%, more preferably at least 99%. The antibodies are antibodies 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2. 1, 116.1, 11.7.1, 12.3.1.1, 12.9.1.1, for heavy and light chains of antibodies selected from 10D1, or for heavy or light chains Even more preferably, the sequence identity or sequence similarity with single is 100%.

In another embodiment, the antibody germline V _kappa A27, germline V _kappa O12, and _(an allele of V H three to thirty-three locus) sequence identity or sequence similarity with the sequence of germline DP50 Is at least 80%, more preferably at least 85%, even more preferably at least 90%, even more preferably at least 94%, more preferably for heavy and light chain full length sequences or for heavy or light chain alone At least 95%, even more preferably at least 99%. It is even more preferred that the antibody has 100% sequence identity or sequence similarity with the germline DP50 heavy chain sequence and / or with the germline A27 or germline O12 light chain sequence.

  In one embodiment, the antibody is directed against antibody 3.1.1, 4.1.1, 4.8.1, 4 for heavy and light chain variable region sequences, or for heavy or light chain variable region sequences alone. .10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 12 .9.1.1 Sequence (eg, amino acid, nucleic acid, or both) identity or sequence similarity with the sequence of 10D1 is at least 80%, more preferably at least 85%, even more preferably at least 90%, Even more preferably at least 94%, preferably at least 95%, more preferably at least 99%. The antibodies are antibodies 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2. 1, 116.1, 11.7.1, 12.3.1.1, 12.2.9.1.1, for heavy and light chain variable region sequences of antibodies selected from 10D1, or heavy chain It is even more preferred that the sequence identity or sequence similarity with the light chain sequence alone is 100%.

In another embodiment, the antibody, for heavy chain variable region sequences, is heavy chain variable sequence of heavy chain germline DP50 (which is an allele of the V _H 3-33 locus) or germline V _κ A27 or germline V. _κ sequence identity or sequence similarity with the light chain variable sequence of O12 is at least 80%, more preferably at least 85%, even more preferably at least 90%, even more preferably at least 94%, more preferably at least 95 %, Even more preferably at least 99%. Even more preferably, the antibody heavy chain region sequence has 100% sequence identity or sequence similarity with the germline DP50 sequence or the germline A27 or germline O12 light chain sequence.

  In one embodiment of the invention, the antibody is antibody 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6. 1.1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 12.9.1.1, 10D1 sequence identity with the FR1-FR4 region sequence Or sequence similarity is at least 80%, more preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, more preferably for FR1-FR4 sequences of heavy chain, light chain, or both Is at least 99%. The antibody is directed to antibody 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4 for FR1-FR4 sequences of heavy chain, light chain, or both. .14.3, 6.1.1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 12.9.1.1, and 10D1 Even more preferably, the identity or sequence similarity is 100%.

  In another embodiment of the invention, the antibody has at least 80%, more preferably at least 85 sequence identity or sequence similarity with the FR1-FR3 region sequence of germline DP50 for the FR1-FR3 heavy chain sequence. %, Even more preferably at least 90%, even more preferably at least 95%, more preferably at least 99%, and most preferably about 100%.

In yet another embodiment of the present invention, the antibody, the light chain sequence of FR1 to FR4, sequence identity or sequence similarity with the FR1 to FR4 region sequences of germline V _kappa A27 or germline V _kappa O12, At least 80%, more preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, more preferably at least 99%, most preferably about 100%.

  In one embodiment of the invention, the antibody is antibody 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6. 1.1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 12.9.1.1, 10D1 heavy chain, light chain, or both CDRs -1, CDR-2, and CDR-3 sequences having at least 80% sequence identity or sequence similarity, more preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, More preferably it is at least 99%. The antibody is directed against antibodies 3.1.1, 4.1.1, 4.8.1, 4.10. For heavy chain, light chain, or both CDR-1, CDR-2, and CDR-3 sequences. 2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 12.9. Even more preferably, the sequence identity or sequence similarity with 1.1 and 10D1 is 100%.

  In another embodiment of the invention, the antibody has at least 80% sequence identity or sequence similarity for the heavy chain CDR-1 and CDR-2 sequences with the germline DP50 CDR-1 and CDR-2 sequences. More preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, more preferably at least 99%, most preferably about 100%.

In yet another embodiment of the present invention, the antibody, the light chain CDR-1, CDR-2, and CDR-3 sequence, CDR-1, CDR-2 germline V _kappa A27 or germline V _kappa O12, And sequence identity or sequence similarity with the CDR-3 sequence is at least 80%, more preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, more preferably at least 99%, Most preferably it is about 100%.

  Examples of antibodies that can be used in the present invention and methods for producing them are, among others, US patent application Ser. No. 09 / 472,087, currently issued as US Pat. No. 6,682,736; International application PCT / US00 / 23356 (WO 01/14424). (Published March 1, 2001) (eg, antibody 10D1, also known as MDX-010, Medarex, Princeton, NJ, USA); International application PCT / US99 / 28739 (published June 8, 2000 as WO00 / 32231) U.S. Pat. Nos. 5,810,097, 5,855,887, 6,051,227, and 6,207,156, each of which is incorporated herein by reference. Information about the amino acid and nucleic acid sequences associated with these antibodies is provided herein, but further information can be found in US Pat. No. 6,682,736 and WO 00/37504, and is described in those applications. Sequences are hereby incorporated herein by reference.

  Certain uses of these antibodies to treat various cancers are discussed in US patent application Ser. No. 10 / 153,382, now published as US Patent Application Publication No. 2003/0086930, which Which is incorporated by reference as if set forth in its entirety herein.

  The properties of human anti-CTLA4 antibodies useful in the methods of the present invention are extensively discussed in, for example, US Pat. No. 6,682,736, including but not limited to antibodies 3.1.1, 4.1.1, 4.8. 4.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 11.6.1, 11.7.1, 12.3.1 .1, 12.9.1.1, 10D1 and other antibodies having the amino acid sequence of the antibody are included. The present invention also relates to antibodies comprising the heavy and light chain CDR amino acid sequences of these antibodies and methods of using antibodies comprising alterations in the CDR regions as described in the applications and patents listed above. The invention also relates to antibodies comprising the heavy and light chain variable regions of those antibodies. In another embodiment, the antibody is antibody 3.1.1, 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1. 1, 11.2.1, 11.6.1, 11.7.1, 12.3.1.1, 12.9.1.1, and 10D1 full-length variable region amino acid sequences, heavy and light chains It is selected from antibodies comprising an amino acid sequence or a CDR amino acid sequence.

  Although the anti-CTLA4 antibodies previously discussed herein may be preferred, those of skill in the art, based on the disclosure provided herein, include a broad class of anti-CTLA4 antibodies, and their identification It should be understood that the present invention is not limited to antibodies. More particularly, human antibodies are preferred, but the invention is in no way limited to human antibodies, but rather the invention includes antibodies that are useful regardless of species origin, among others, chimeric antibodies, humanized antibodies, and / or Or a primatized antibody. In addition, the antibodies exemplified herein were obtained using a transgenic mammal, for example, a mouse containing a human immune repertoire, and those skilled in the art will be able to describe the disclosure provided herein. On the basis, it should be understood that the present invention is not limited to antibodies produced by this method or any other specific method. Instead, the present invention includes, but is not limited to, methods known in the art (eg, screening of phage display libraries, etc.) or methods developed in the future to generate anti-CTLA4 antibodies of the present invention. An anti-CTLA4 antibody produced by any method. Based on the present disclosure and the extensive disclosure provided, for example, in Hanson et al. US Pat. No. 6,682,736, US Patent Application Publication No. 2002/0088014, those skilled in the art will Antibodies that are useful in the treatment of breast cancer in combination with hormonal therapeutic agents can be readily generated and identified.

  The present invention relates to Hanson et al. S. As disclosed in US Pat. No. 6,682,736, non-human transgenic mammals, ie, human antibodies produced using XenoMouse ™ (Abgenix, Inc., Fremont, Calif., USA) are included.

  Another transgenic mouse system for the production of “human” antibodies is called “HuMAb-Mouse ™” (Medarex, Princeton, NJ, USA), and unrearranged human heavy chains (μ and γ) and Includes targeted mutations that inactivate endogenous mu and kappa chain loci along with human immunoglobulin gene loci that encode immunoglobulin sequences for kappa light chains (Lonberg et al., Nature 368: 856-859 (1994) and US Pat. No. 5,770,429).

  However, the present invention is not so limited, for example, Tomizuka et al., Proc Natl Acad Sci USA 97: 722 (2000); Kuroiwa et al., Nature Biotechnol 18: 1086 (2000); Mikayama et al. Kirin TC Mouse (TM) (Kirin Brewery, Tokyo); HuMAb-Mouse (TM) (Medarex, Princeton, NJ); XenoMouse (TM) described in US Patent Application Publication No. 2004/0120948 Human anti-CTLA4 antibodies produced using any transgenic mammal are used, such as Abgenix, Inc., Fremont, Calif., Supra. Thus, the present invention includes the use of anti-CTLA4 antibodies produced using any transgenic animal or other non-human animal.

  In another embodiment, the antibodies used in the methods of the invention are “humanized” rather than fully human. In particular, murine antibodies or other species of antibodies can be “humanized” or “primatized” using techniques well known in the art. See, for example, Winter and Harris, Immunol. Today 14: 43-46 (1993), Wright et al., Crit. Reviews in Immunol. Vol. 12: 125-168 (1992), Cabilly et al., U.S. Pat. No. 4,816,567, Mage and Lamoyi, “Monoclonal Antibody Production Techniques and Applications”, pages 79-97, Marcel Inc. See, New York, New York (1987). Thus, obtained from any species, including humanized antibodies, chimeric antibodies (eg, single chain antibodies obtained from camelids such as those described in US Pat. Anti-CTLA4 antibodies, as well as any human antibody, can be combined with therapeutic agents to practice the novel methods disclosed herein.

  As will be understood based on the disclosure provided herein, antibodies for use in the present invention can be obtained from non-human transgenic mammals and hybridomas derived therefrom, but also in cell lines other than hybridomas. Can be expressed.

  Mammalian cell lines that can be used as hosts for expression are well known in the art and include, but are not limited to, Chinese hamster ovary (CHO) cells, NS0, Sp2, HEK, HeLa cells, pup hamster kidney (BHK). This includes a number of immortal cell lines available from the American Cultured Cell Line Conservation Agency (ATCC), including cells, monkey kidney cells (COS), and human liver cancer cells (eg, Hep G2). Non-mammalian prokaryotic and eukaryotic cells can also be used, including bacterial, yeast, insect, and plant cells.

  For example, but not limited to: Sambrook and Russell, “Molecular Cloning, A Laboratory Approach, Cold Spring Harbor Press”, Cold Spring Harbor, New York, USA (2001), Ausubel et al., “Current Protocol MolesleSole A variety of expression systems well known in the art can be used, such as those described in New York, USA (2002). These expression systems include, among many others, systems based primarily on dihydrofolate reductase (DHFR). The expressed glutamine synthetase system is discussed in whole or in part in connection with European Patent Nos. 0216846B1, 0256605B1, and 0323997B1, and European Patent Application No. EP89303964. In one embodiment, the antibodies used are made using the glutamine synthetase system (GS-NS0) in NS0 cells. In another embodiment, the antibody is made using the DHFR system in CHO cells. Both systems are well known in the art and are described, among others, in Barnes et al., Biotech & Bioengineering 73: 261-270 (2001) and references cited therein.

  Where site-directed mutagenicity of antibody CH2 domains to eliminate glycosylation is preferred to prevent any changes in immunogenicity, pharmacokinetics, and / or effector function that occur as a result of non-human glycosylation There is. In addition, antibodies can be produced by enzymatic methods (see, eg, Thotakura et al., Meth. Enzymol. 138: 350 (1987)) and / or chemical methods (eg, Hakimudin et al., Arch. Biochem. Biophys., 259: 52 (1987)) can be deglycosylated.

  Furthermore, the present invention includes the use of anti-CTLA4 antibodies comprising altered glycosylation patterns. One of ordinary skill in the art will appreciate that based on the disclosure provided herein, anti-CTLA4 antibodies can be modified to include additional, fewer, or different glycosylation sites than naturally occurring antibodies. is there. Such modifications are described, for example, in U.S. Patent Application Publication Nos. 2003/0207336, 2003/0157108, International Patent Publications WO 01/81405, and 00/24893.

  Furthermore, the present invention includes the use of anti-CTLA4 antibodies, regardless of any glycoform present on the antibody. Moreover, methods for extensively reconstructing glycoforms present on glycoproteins are well known in the art, for example, International Patent Publications WO 03/031464, WO 98/58964, and WO 99/22764, US patent applications. Publication Nos. 2004/0063911, 2004/0132640, 2004/0142856, 2004/0072290, and Umana et al. US Pat. No. 6,602,684.

  Further, the present invention is not limited thereto, for example, U.S. Patent Application Publication Nos. 2003/0207346 and 2004/0132640, U.S. Pat. Nos. 4,640,835, 4,496,689, 4,301,144, 4,670,417, In the art, including those wherein the polypeptide is linked to one of a variety of non-proteinaceous polymers, such as polyethylene glycol, polypropylene glycol, or polyoxyalkylene, as described in US Pat. Nos. 4,791,192 and 4,179,337. Including the use of anti-CTLA4 antibodies with any modification by known covalent and non-covalent bonds.

  The present invention also includes the use of anti-CTLA4 antibodies or antigen-binding portions thereof, chimeric proteins comprising, for example, human serum albumin polypeptides or fragments thereof. Whether a chimeric protein is produced using recombinant methods, for example by cloning of a chimeric nucleic acid encoding the chimeric protein, or by chemical linking of two peptide moieties, as used herein. Those skilled in the art who have the teachings provided will be familiar with such chimeric proteins, including, but not limited to, such as increased stability and increased serum half-life for the antibodies of the present invention. It is to be understood that such molecules are included herein.

  Antibodies produced for use in the present invention need not initially have the particular isotype desired. Rather, the antibody produced can have any isotype and can then be isotyped using conventional techniques. These include direct recombination techniques (see, eg, US Pat. No. 4,816,397) and cell-cell fusion techniques (see, eg, US Pat. No. 5,916,771).

  The effector function of the antibodies used in the present invention can be altered for various therapeutic applications by isotype switching to IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM. Moreover, it is possible to avoid relying on complement for cell death, for example by using bispecific antibodies, immunotoxins, or radiolabels.

  Antibodies 4.1.1, 4.13.1, and 11.2.1 are IgG2 antibodies, and the variable region sequences of this antibody are described herein (FIGS. 1-3), as well as references herein. The full-length sequences of these antibodies, as well as those provided in the incorporated applications and patents, as discussed more fully elsewhere herein, as well as SEQ ID NOs: 1-36 It is understood that the use of any antibody comprising the described sequence and further comprising any constant region is included herein. Similarly, any antibody comprising a full length sequence of 10D1, or any portion thereof comprising a sequence encoding an antigen binding portion of 10D1, can be used in accordance with the methods of the invention.

  Accordingly, those of ordinary skill in the art who have received the teachings provided herein will readily appreciate that the anti-CTLA4 antibody-therapeutic agent combinations of the present invention can include multiple types of anti-CTLA4 antibodies.

  Furthermore, one of ordinary skill in the art, based on the disclosure provided herein, is not limited to administration of a single antibody alone, but rather at least one anti-CTLA4 antibody, eg, 4.1.1, It should be understood that this includes administration of one of 4.13.1, or 11.2.1 in combination with a therapeutic agent. Furthermore, any combination of anti-CLTA4 antibodies can be combined with at least one therapeutic agent, and the present invention includes any such combinations and combinations thereof.

IV. The present invention relates to administering a combination of an anti-CTLA4 antibody and an aromatase inhibitor to a patient for the treatment of breast cancer.

  Hormonal therapeutic agents for the treatment of breast cancer tumors that express the estrogen receptor are well known in the art. These hormone therapy agents include, but are not limited to, tamoxifen, which binds to the receptor and thereby inhibits the receptor: ligand interaction, and a receptor that can be used to mediate signal transduction through the receptor: ligand interaction. Included are compounds that affect the receptor itself, such as fulvestrant, which degrades the receptor to a lesser extent.

  Hormonal therapeutic agents include, but are not limited to, inhibitors of aromatase enzymes such as anastrozole, letrozole, and exemestane. These anti-hormonal agents, unlike tamoxifen and fulvestrant described above, reduce the level of ligand available for binding to the receptor and inhibit receptor: ligand interactions. More specifically, aromatase inhibitors inhibit the aromatization of the A-ring of androgen substrate for estrogen production, a necessary step in the biosynthesis of receptor ligands, ie estrogens. Thus, aromatase inhibitors mediate the reduction of estrogen serum levels, thereby inhibiting the growth of tumor cells that require hormones for growth.

  Non-steroidal aromatase inhibitors (“AI”), such as, but not limited to, anastrozole and letrozole, reversibly bind to the heme portion of the aromatase molecule and are associated with increased aromatase activity. This is especially associated with the development of tumor resistance associated with long-term use of inhibitors. Furthermore, since this inhibitor binds reversibly to the enzyme, in the presence of these inhibitors, estrogen biosynthesis occurs without the need for the synthesis of a new aromatase (Miller, Clin Breast Cancer Volume 1: S9- (See page S14 (2000)), which may provide another opportunity for tumor cells to avoid inhibition.

  Steroidal aromatase inhibitors such as exemestane differ from nonsteroidal inhibitors (eg, anastrozole or letrozole) in that asteroid drugs bind irreversibly to the enzyme's substrate binding site. Associated with a decline in For example, exemestane is structurally related to the natural substrate androstenedione and is recognized by the enzyme as a fake substrate, so it competes with the natural substrate at the active site. Exemestane is then converted to an intermediate that irreversibly binds to the enzyme while causing its inactivation (ie, “suicide inhibition”). For example, U.S. Patent Application No. 10/611653 published as U.S. Patent Application Publication No. 2004/0082557, U.S. Patent Application No. 10/363935 published as U.S. Patent Application Publication No. 2004/0024044, and See U.S. Patent Application No. 10/343595, published as Patent Application Publication No. 2003/0158168.

  That is, unlike reversible non-steroidal inhibitors, exemestane is a steroidal aromatase inactivator that irreversibly binds to aromatase and causes almost complete suppression of aromatase activity not only in peripheral tissues but also in tumors. Yes, mediates a substantial reduction in serum estrogen levels without exogenous endocrinological effects (Kaufmann et al., J Clin Oncol 18: 1399-1411 (2000)). Exemestane is currently approved in the United States, Canada, and Europe for second-line treatment of women with postmenopausal metastatic breast cancer (eg, when the disease has progressed after tamoxifen first-line therapy). Exemestane has a response rate of about 15%, a median survival of 123.4 weeks, and lack of cross-resistance with non-steroidal drugs, so it may be effective after anastrozole and letrozole treatment (Geisler et al., Clin Cancer Res 4: 2089-2093 (1998); Lonning, J Clin Oncol 18: 2234-2244 (2000)). More recently, exemestane has been approved in Europe for adjuvant treatment of estrogen receptor positive invasive early breast cancer after 2-3 years of initial adjuvant tamoxifen therapy in postmenopausal women.

  Unlike its non-steroidal aromatase inhibitors, exemestane further prevents the biosynthesis of estrogens that require a novel synthesis of the aromatase enzyme because its intermediates bind irreversibly. Moreover, the different mechanisms of action of non-steroidal and steroidal aromatase inhibitors can explain the phenomenon that one aromatase inhibitor can become effective after the other fails. More specifically, treatment of postmenopausal breast cancer patients who were treated with exemestate after failure of both tamoxifen and non-steroidal aromatase inhibitors provided clinical benefits (Lonning et al., J Clin Oncol. 18: 2234-2244 (2000)).

  Thus, anti-CTLA4 antibodies in combination with any aromatase inhibitor or inhibitors can be used to treat breast cancer patients whose tumors express estrogen receptor (ER +). Furthermore, even though the present invention is exemplified by a combination comprising exemestane, the present invention is in no way limited to this inhibitor or its inhibition mechanism. In fact, it is not necessary to know the mechanism by which an aromatase inhibitor inhibits an enzyme. That is, any inhibitor that can detectably inhibit aromatization of the A ring of the substrate in estrogen biosynthesis using any known assay to assess its activity is a combination of the present invention. Can be used inside.

V. Additional Agents The present invention can be further combined with additional agents and therapies such as chemotherapy, surgery, radiation therapy, transplantation, etc. to treat patients. That is, patients may include, but are not limited to, growth factor inhibitors, biological response modifiers, alkylating agents, intervening antibiotics, vinca alkaloids, immunomodulators, taxanes, but not limited to lasofoxifene Additional chemotherapy with well-known drugs, such as selective estrogen receptor modulators (SERMs), and angiogenesis inhibitors can be received.

  There are many therapeutic agents, among others many are described in, for example, US Patent Application Publication Nos. 2004/0005318, 2003/0086930, 2002/0086014, and International Publication No. WO 03/086459, Is incorporated herein by reference in its entirety. Such therapeutic agents include, but are not limited to, topoisomerase I inhibitors; other antibodies (rituximab, bevacizumab, trastuzumab, anti-IGF 1R antibodies [eg CP-751,871], anti-CD40, among many others. Chemotherapeutic agents such as imatinib (GLEEVEC), SU11248 (SUTENT; sunitinib), SU12662, SU14813; BAY43-9006, AG-013736 (axitinib) ), Toll-like receptor stimulants (eg, but not limited to TLR-9 agonists such as CPG-7909, also referred to as PF03512676 or PROMUNE), indoleamine-2,3, -dioxygenase (IDO) Inhibitors; Selective estrogen receptor modulators (SERMs such as lasofoxifene); taxanes; vinca alkaloids; temozolomides; angiogenesis inhibitors; EGFR inhibitors; VEGF inhibitors; erbB2 receptor inhibitors; For example, farnesyl protein transferase inhibitors, and αvβ3 inhibitors, αvβ5 inhibitors, p53 inhibitors, etc.); immunomodulators; biological response modulators; cytokines; tumor vaccines; tumor-specific antigens; Dendritic cell and stem cell therapy; alkylating agents; folic acid antagonists; pyrimidine antagonists; anthracycline antibiotics; platinum compounds; immune costimulatory molecules (eg, CD4, CD25, PD-1, B7-H3) 4-1BB, cow 40, ICOS, CD30 HLA-DR, include MHCII, and LFA), and its agonist antibody.

  In one embodiment, the methods of the invention can be further combined with transplantation, such as stem cell transplantation, to provide a therapeutic benefit to breast cancer patients. Stem cell transplantation can be performed according to methods known in the art and can be allogeneic or autologous stem cell transplantation. Those skilled in the art will also be aware that, based on the disclosure provided herein, transplantation involves adoptive transfer of either lymphocytes obtained from autologous or HLA-matched donors. If the method involves stem cell transplantation, the first dose of the antibody-AI therapeutic agent combination can be administered after the mammal's immune system has recovered from transplantation, for example, 1-12 months after transplantation. . In certain embodiments, the first dose is administered 1 to 3 months, or 1 to 4 months after transplantation. Transplantation methods include “Appelbaum in Harrison's Principles of Internal Medicine”, Chapter 14, Braunwald et al., 15th edition, McGraw-Hill Professional (2001), which is incorporated herein by reference. (Treatises).

  As previously pointed out herein, there are many chemotherapeutic agents currently available for the treatment of tumors that are suitable for use in the combination therapy of the present invention. For example, alkylating agents are a class of drugs that alkylate DNA while limiting stranding and replication. A preferred alkylating agent for use in the method of the present invention is cyclophosphamide (CYTOXAN).

  Folate antagonists bind to dihydrofolate reductase (DHFR) and prevent pyrimidine (thymidine) synthesis. Methotrexate and pemetrexed (ALIMTA) are folic acid antagonists suitable for use in the methods of the invention. In addition to DHFR, pemetrexed also inhibits two other folate-dependent enzymes involved in thymidine synthesis, thymidylate synthase and glycinamide ribonucleotide formyltransferase.

  Pyrimidine antagonists inhibit enzymes involved in pyrimidine synthesis. As a pyrimidine analog, this antagonist also prevents DNA production by competing with normal nucleotides for incorporation into the DNA molecule. Pyrimidine antagonists suitable for use in the methods of the present invention include 5-fluorouracil (5-FU); capecitabine (XELODA), a 5′-deoxy-5-fluorouridine that is converted to 5-FU by an enzyme in vivo. Prodrugs of (5'-FDUR); and gemcitabine (GEMZAR).

  Anthracycline antibiotics intervene between DNA strands to suppress unraveling of DNA. Anthracycline antibiotics include doxorubicin hydrochloride (adriamycin), epirubicin hydrochloride (ELLENCE, PHARMORUBICIN), daunorubicin (CERUBIDINE, DAUNOXOME), and idarubicin hydrochloride (IDAMYCIN PFS, ZAVEDOS). Preferred anthracyclines for use in the present invention include doxorubicin and epirubicin.

  A platinum compound exerts its antitumor effect by intervening and inserting between DNA strands that suppress unraveling of DNA. Platinum compounds useful in the methods of the present invention include cisplatin (PLATINOL), oxaliplatin (ELOXATIN), and carboplatin (PARAPLATIN).

  Taxanes promote the assembly of microtubules while inhibiting their degradation to tubulin, thereby blocking the ability of cells to break the spindle during mitosis. Taxanes have demonstrated significant activity against many solid tumors as single drug therapy and in combination with other chemotherapeutic agents. One embodiment of the combination therapy of the invention involves the use of one or more taxanes in combination with an IGF-1R antibody. Taxanes suitable for use in combination with an IGF-1R antibody include docetaxel (TAXOTERE) and paclitaxel (TAXOL).

  Taxane derivatives that may have activity in cells that are resistant to doxorubicin, vinblastine, paclitaxel, docetaxel, etc. include XRP-9981 (Sanofi Aventis), which are included in the present invention.

  Vinca alkaloids such as taxanes are “spindle toxins” that act on the microtubules that form the spindle. They inhibit mitosis by preventing microtubule assembly while preventing the formation of spindles. Vinca alkaloids include vindesine (ELDISINE), vinblastine sulfate (VELBAN), vincristine sulfate (ONCOVIN), and vinorelbine tartrate (NAVELBINE). A preferred vinca alkaloid for use in the method of the present invention is vinorelbine.

  BMS-247550 (ixabepilone) promotes tubulin polymerization and microtubule stabilization, thereby blocking cells in the G2-M phase and inducing tumor cell apoptosis. This drug is active against taxane-resistant cells.

  Rapamycin analogs that bind and inhibit the mammalian rapamycin target protein (mTOR) are also useful, including, among others, CCI-779 (Temsilolimus, Wyeth) and RAD-001 (Everolimus, CERTICAN, Novartis).

  Camptothecin analogs act through inhibition of topoisomerase I, an enzyme critical for DNA replication and packaging. Tumor cells have higher topoisomerase I levels than normal tissues. A camptothecin analog useful in the method of the present invention is irinotecan (CAMPTOSAR).

  In certain embodiments of the invention, the method described above is combined with a cancer vaccine. See, for example, Oh et al., Cancer Res. 64: 2610-2618 (2004) (TARP epitopes and breast cancer); Kontani et al., Int J Molec Med 12: 493-502 (2003) (dendritic cell vaccine targeting MUC1 mucin) Holberg and Sandmeier, Expert Rev Vaccines Volume 3: 269-277 (2004) (Vaccination for Breast or Ovarian Cancer); Thus, useful vaccines include, but are not limited to, breast cancer associated antigens (eg, HER-2 / neu, mammoglobulin, prostate and breast tumor associated protein [TARP], MUC1, CEA, sialyl-Tn, And other carbohydrate antigens), other tumor cancer-associated antigens (eg, p53, telomerase), those consisting of anti-idiotype antibodies such as 11D10, and vaccines containing GM-CSF, vaccines based on DNA and cells, trees Dendritic cell vaccines, recombinant virus (eg vaccinia virus) vaccines, and heat shock protein (HSP) vaccines (eg HSPPC-96, Antigens Inc.). Useful vaccines include tumor vaccines such as those formed with breast tumor cells, useful vaccines can be autologous or allogeneic, and can be based on peptides, DNA, or cells. What to do.

  The vaccine can be administered before or after administration of the antibody-aromatase inhibitor combination, and when chemotherapy is part of the dosage regimen, the vaccine can be administered before chemotherapy. In certain embodiments, the antibody-aromatase inhibitor combination of the invention may be administered prior to chemotherapy. In yet other embodiments, treatment can be combined with stem cell transplantation. That is, the antibody-aromatase inhibitor combination can be administered before or after stem cell transplantation. The vaccine may be administered before or after stem cell transplantation, in certain embodiments, simultaneously with the antibody.

  The above-mentioned treatment includes drugs that suppress EGFR (epidermal growth factor receptor) response, such as EGFR antibodies, EGF antibodies, EGFR inhibitor molecules; VEGF (blood vessels such as VEGF receptors and molecules that can inhibit VEGF) Endothelial growth factor) inhibitors; and signaling inhibitors such as erbB2 receptor (HER2) inhibitors such as organic molecules and antibodies that bind to the erbB2 receptor, such as trastuzumab (Herceptin, Genentech, Inc., San Francisco, Calif., USA) ), HER dimerization inhibitor (HDI) pertuzumab (2C4, OMNITARG, Genentech).

  EGFR inhibitors are described, for example, in International Patent Publications WO95 / 19970, WO98 / 14451, WO98 / 02434, and US Pat. No. 5,747,498, and such substances are used in the present invention as described herein. can do. Agents that inhibit EGFR include, but are not limited to, monoclonal antibody C225, anti-EGFR22 Mab (ImClone Systems Inc., New York, USA), and ABX-EGF (Abgenix Inc., remont, CA, USA), compound ZD -1839 (AstraZeneca), BIBX-1382 (Boehringer Ingelheim), MDX-447 (Medarex, Inc., Annedale, NJ, USA), and OLX-103 (Merck & Co., Whitehouse VCT, St. CV, USA) Research) and EGF fusion toxin (Seragen Inc., Country Maryland Hopkinton) is included. These agents and other agents that inhibit EGFR can be used in the present invention.

  Compounds commissioned to inhibit epidermal growth factor receptor (EGFR) tyrosine kinase (TK) are a relatively new class of antineoplastic agents useful in the methods of the invention. Many human cancers express EGFR family members on the cell surface. Ligand, when bound to EGFR, triggers a cellular reaction cascade that leads to increased cell division and other aspects of cancer development and progression, including angiogenesis, metastatic spread, and inhibition of apoptosis. affect. EGFR-TK inhibitors are members of the EGFR family (EGFR (also known as HER1 or ErbB-1), HER2 / neu (also known as ErbB-2), HER3 (also known as ErbB-3). Or HER4 (also known as ErbB-4)) may be selectively targeted, or two or more of these may be targeted. EGFR-TK inhibitors suitable for use in the present invention include gefitinib (IRESSA), erlotinib (TARCEVA), CI-1033 (Pfizer), GW2016 (GlaxoSmithKline), EKB-569 (Wyeth), PKI-166 (Novartis) CP-724, 714 (Pfizer), and BIBX-1382 (Boeringer-Ingelheim). Additional EGFR-TK inhibitors are described in US Pat. No. 6,890,924.

  VEGF inhibitors such as SU-5416, SU-6668, SU-11248, SU-12626, SU-14683 (Sugen Inc., San Francisco, Calif.), And AG-013736 (Pfizer) are also used in combination with antibodies. be able to. Examples of VEGF inhibitors include international patent application PCT / IB99 / 00797 (filed on May 3, 1999), international patent publications WO99 / 24440, WO95 / 21613, WO99 / 61422, WO98 / 50356, WO99 / 10349, WO97 / 32856, WO 97/22596, WO 98/54093, WO 98/02438, WO 99/16755, WO 98/02437, U.S. Pat. Other examples of some detailed VEGF inhibitors useful in the present invention are IM862 (Cytran Inc., Kirkland, WA, USA), Genentech, Inc. , IMC-1C11 Imclone antibody, an anti-VEGF monoclonal antibody in San Francisco, California, USA, and angiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colorado, USA) and Chiron (Emeryville, CA, USA).

  GW-282974, GW-572016 (Lapatinib) (Glaxo Wellcome plc) and the monoclonal antibody AR-209 (Aronex Pharmaceuticals Inc., Woodlands, Texas, USA), Trastuzumab (HERCEPTIN, San Francisco, USA) ErbB2 receptor inhibitors such as pertuzumab (OMNITARG, 2C4, Genentech, HER2 dimerization inhibitor HDI), and 2B-1 (Chiron) can be combined with antibody-aromatase inhibitor combination therapy. Other erbB2 receptor inhibitors are described, for example, in International Patent Publications WO 98/02434, WO 99/35146, WO 99/35132, WO 98/02437, WO 97/13760, WO 95/19970, US Pat. Nos. 5,587,458, and 5,877,305. Are listed. The erbB2 receptor inhibitors useful in the present invention are also described in EP1029853 (published on August 23, 2000) and international patent publication WO00 / 44728 (published on August 3, 2000). The erbB2 receptor inhibitor compounds and substances described in the above-mentioned PCT applications, US patents, and US provisional applications, as well as other compounds and substances that inhibit the erbB2 receptor, include the antibodies and aromatase inhibitors of the invention. Can be used with combinations.

  The treatments of the present invention also include, but are not limited to, other agents that can enhance an anti-tumor immune response, such as additional different CTLA4 antibodies; other agents that can also block CTLA4; farnesyl protein Anti-proliferative agents such as transferase inhibitors; αvβ3 inhibitors such as αvβ3 antibody VITAXIN; vβ5 inhibitors; p53 inhibitors and the like are used in conjunction with other agents useful in treating abnormal cell growth or cancer.

  When the antibody of the present invention is administered in combination with another immunomodulator, the immunomodulator is, for example, a dendritic cell activator such as a CD40 ligand or an anti-CD40 agonist antibody, as well as an antigen-presenting activator, T cell-directing agent. It can be selected from the group consisting of sex activators, inhibitors of immunosuppressive factors associated with tumors such as TGF-β (transforming growth factor β) and IL-10. Preferred anti-CD40 agonist antibodies are international patent application PCT / US02 / 36107 filed on Nov. 8, 2002 and currently published as International Patent Publication WO 03/040170, and filed Nov. 8, 2002 and currently US patents. Including, but not limited to, the antibodies disclosed in US patent application Ser. No. 10 / 292,880, published as published US 2003/0211100. H-A78T, 3.1.1H-A78T-V88A-V97A, 3.1.1L-L4M-L83V, 3.1.1H-A78T-V88A-V97A / 3.1.1L-L4M-L83V, 7. 1.2, 10.8.3, 15.1.1, 21.2.1, 21.4.1, 21.2.1, 22.1.1.1H-C109A, 23.5.1, 23. This includes antibodies having heavy and light chain amino acid sequences of 25.1, 233.28.1, 233.28.1H-D16E, 23.29.1, and 24.2.1.

  This therapeutic regimen may be combined with antibodies or other ligands that bind to IGF-1R (insulin-like growth factor 1 receptor) and inhibit tumor growth. Detailed anti-IGF-1R antibodies that can be used in the present invention include International Patent Application PCT / US01 / 51113 published in International Patent Publication No. WO 02/053596 filed 12/20/01, 2004 8 Examples include those described in International Patent Application PCT / IB2004 / 002555 filed on May 3 and published as International Patent Publication WO2005 / 016967. Preferred anti-IGFR-1R antibodies are, for example, the heavy chains of antibodies 2.12.1, 2.13.2, 2.14.3, 3.1.1, 4.9.2, and 4.17.3. And antibodies having a light chain amino acid sequence.

  The antibody of the present invention may be administered together with cytokines such as IL-2, interferon (eg, IFN-γ, IFN-α, etc.), GM-CSF, IL-12, IL-18, and FLT-3L.

  The therapeutic regimes described herein are MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, COX-II, which can be used with antibodies in the methods of the invention. It can be combined with anti-angiogenic agents such as (cyclooxygenase II) inhibitors. Examples of useful COX-II inhibitors include CELEBREX (celecoxib), valdecoxib, and rofecoxib. Examples of useful matrix metalloprotease inhibitors are International Patent Publications WO96 / 33172, WO96 / 27583, WO98 / 07697, WO98 / 03516, WO98 / 34918, WO98 / 34915, WO98 / 33768, WO98 / 30566, WO90 / 05719, WO99 / 52910, WO99 / 52889, WO99 / 29667, European Patent Application No. 780386 (published on June 25, 1997), No. 97304971 (filed on July 8, 1997), No. 99308617 (1999 October 10) No. 606046 (published July 13, 1994), No. 931788 (published Jul. 28, 1999), No. 99302232 (filed Mar. 25, 1999), International application PCT / IB98 / 01 13 (filed July 21, 1998), British Patent Application No. 99129961 (filed June 3, 1999), US Provisional Application No. 60/148464 (filed August 12, 1999), and US Patent No. No. 5863949 and No. 5861510.

  Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity inhibiting MMP-1. More preferred are other matrix-metalloproteinases (ie, MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-2 and / or MMP-9 is selectively inhibited as compared with MMP-12 and MMP-13).

Some detailed examples of MMP inhibitors useful in the present invention are AG-3340, RO32-3555, RS13-0830, and the compounds listed in the following list. That is,
3-[[4- (4-Fluoro-phenoxy) -benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl) -amino] -propanoic acid,
3-exo-3- [4- (4-fluoro-phenoxy) -benzenesulfonylamino] -8-oxa-bicyclo [3.2.1] octane-3-carboxylic acid hydroxyamide,
(2R, 3R) 1- [4- (2-chloro-4-fluoro-benzyloxy) -benzenesulfonyl] -3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide,
4- [4- (4-fluoro-phenoxy) -benzenesulfonylamino] -tetrahydro-pyran-4-carboxylic acid hydroxyamide,
3-[[4- (4-Fluoro-phenoxy) -benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl) -amino] -propanoic acid,
4- [4- (4-chloro-phenoxy) -benzenesulfonylamino] -tetrahydro-pyran-4-carboxylic acid hydroxyamide,
(R) 3- [4- (4-Chloro-phenoxy) -benzenesulfonylamino] -tetrahydro-pyran-3-carboxylic acid hydroxyamide,
(2R, 3R) 1- [4- (4-fluoro-2-methyl-benzyloxy) -benzenesulfonyl] -3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide,
3-[[4- (4-Fluoro-phenoxy) -benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-ethyl) -amino] -propanoic acid,
3-[[4- (4-Fluoro-phenoxy) -benzenesulfonyl]-(4-hydroxycarbamoyl-tetrahydro-pyran-4-yl) -amino] -propanoic acid,
3-exo-3- [4- (4-chloro-phenoxy) -benzenesulfonylamino] -8-oxa-bicyclo [3.2.1] octane-3-carboxylic acid hydroxyamide,
3-endo-3- [4- (4-fluoro-phenoxy) -benzenesulfonylamino] -8-oxa-bicyclo [3.2.1] octane-3-carboxylic acid hydroxyamide,
(R) 3- [4- (4-Fluoro-phenoxy) -benzenesulfonylamino] -tetrahydro-furan-3-carboxylic acid hydroxyamide,
And pharmaceutically acceptable salts, and solvates of said compounds.

  Radiation therapy can be administered according to well-known radiotherapy methods for breast cancer treatment. Radiation treatment doses and treatment plans can be readily determined by one skilled in the art and are based on the stage of the disease and other factors well known in the art.

  Co-administration (combination therapy) of an antibody with an aromatase inhibitor includes administering a pharmaceutical composition comprising both an anti-CTLA4 antibody and one or more aromatase inhibitors, and one comprising an anti-CTLA4 antibody and the other comprising Administering two or more separate pharmaceutical compositions comprising an aromatase inhibitor. In addition, co-administration or combination (joint) therapy generally means that the antibody and the additional therapeutic agent are administered simultaneously with each other, although simultaneous, sequential, or separate dosing of the individual components of the treatment is also possible. Including. It will also be appreciated that when the antibody is administered intravenously and the aromatase inhibitor is administered orally (eg, exemestane), the combination is preferably administered as two separate pharmaceutical compositions.

  In addition to the first and second components, the present invention also includes the administration of other therapeutic agents at the same time as one or more of these components, or prior and / or subsequent administration. Such therapeutic agents include cancer vaccines, anti-vascular drugs, anti-proliferative drugs, and to provide supportive care such as but not limited to analgesics, antiemetics, anti-diarrheal drugs, and steroids. Symptomatic drugs are included. Preferred antiemetics include ondansetron hydrochloride, granisetron hydrochloride, and metoclopramide. Preferred antidiarrheal drugs include diphenoxylate and atropine (LOMOTIL), loperamide (IMMODIUM), octreotide (SANDOSTATIN), olsalazine (DIPENTUM), and mesalamine (ASACOL). Preferred steroids include the non-absorbable steroid budesonide (ENTOCORT) and steroids for systemic administration, dexamethasone (DECADRAN) and prednisone (METICORTEN).

  The duration of each administration can vary from rapid administration to continuous perfusion. As a result, for the purposes of the present invention, combinations are not limited solely to those obtained by physical association of components, but allow for separate administration that may be contemporaneous or spaced apart during the period. But there is. The composition according to the present invention is preferably a composition that can be administered parenterally. However, in the case of localized topical therapy, these compositions may be administered orally or intraperitoneally.

  As will be appreciated by those skilled in the art, the choice of therapeutic agent to be used in combination with anti-CTLA4 antibody and aromatase inhibitor combination therapy, and the timing of its use, depends in part on the type and stage of the cancer being treated. It is determined. For example, early breast cancer (when the cancer has not spread outside the breast) is generally followed by surgery and radiation followed by adjuvant chemotherapy or hormone hormone therapy, although any of the therapies of the present invention is anti-CTLA4. It can be combined with an antibody-aromatase inhibitor combination. Typical adjuvant chemotherapy for early breast cancer includes cyclophosphamide, methotrexate, and 5-FU (“CMF”); 5-FU, doxorubicin, and cyclophosphamide (“FAC”); docetaxel, Doxorubicin, and cyclophosphamide ("TAC"); doxorubicin and cyclophosphamide ("AC"); doxorubicin and cyclophosphamide, followed by paclitaxel ("AC and T"); and 5-FU, epirubicin, And cyclophosphamide ("FEC"). As previously discussed elsewhere herein, tamoxifen is an antihormonal treatment administered at this stage. That is, the antibody-aromatase inhibitor combination can be administered in combination with an additional hormone therapy (eg, another aromatase inhibitor tamoxifen, fulvestrant, or any combination thereof) Can be co-administered in combination with adjuvant chemotherapy (eg, CMF, FAC, TAC, AC, AC and T, and / or FEC, or chemotherapeutic agent, in combination with antibody-aromatase inhibitor combination, among others) Can be administered individually).

  In locally advanced breast cancer where the cancer has spread only to nearby tissues or lymph nodes, patients often receive chemotherapy prior to surgery and radiation, followed by adjuvant hormone therapy. Alternatively, surgery / radiation is followed by adjuvant chemotherapy followed by adjuvant hormone therapy. The anti-CTLA4 antibody-aromatase inhibitor combination may be used before or after surgery / radiation, whether it is a chemotherapeutic agent or an additional hormone therapy agent (eg, another aromatase inhibitor, tamoxifen, fulvestrant Or any combination thereof). Typical chemotherapy regimens for locally advanced breast cancer include FAC, AC, FEC, and doxorubicin plus docetaxel (“AT”).

  Metastatic breast cancer has spread from the starting breast to other parts of the body. Chemotherapy may optionally be placed before hormone therapy. First line hormone therapy currently includes tamoxifen and anastrozole. First line chemotherapy treatment plans currently include FAC, TAC, docetaxel plus epirubicin, docetaxel, paclitaxel, capecitabine, vinorelbine, and trastuzumab (HERCEPTIN). Second line treatment includes docetaxel, either alone or in combination with capecitabine. The method of the present invention is suitable for use as both first line therapy and second line therapy. Furthermore, the methods of the invention can be combined with any combination of radiation therapy and stem cell transplantation and any of the treatments described herein known in the art or developed in the future.

  In one embodiment, the additional therapeutic agent administered in addition to antibody-aromatase inhibitor therapy is an alkylating agent. In one aspect, the alkylating agent is cyclophosphamide.

  In another embodiment, the additional agent is a folic acid antagonist. In one aspect, the folic acid antagonist is selected from the group consisting of methotrexate and pemetrexed.

  In one embodiment of the invention, the additional agent is a pyrimidine antagonist. The pyrimidine analog may be selected from the group consisting of 5-FU, capecitabine and gemcitabine.

  In another embodiment of the invention, the additional agent is an anthracycline antibiotic. In one aspect, the anthracycline antibiotic is selected from the group consisting of epirubicin and doxorubicin. In another embodiment, the anthracycline antibiotic is doxorubicin.

  In another embodiment of the invention, the additional agent is a platinum compound. The platinum compound may be selected from the group consisting of cisplatin and carboplatin.

  In yet another embodiment of the invention, the additional agent is a taxane. In one aspect, the taxane is docetaxel and in another aspect, the taxane is paclitaxel.

  In one embodiment of the invention, the anti-CTLA4 antibody and aromatase inhibitor combination is administered with docetaxel in further combination with at least one of capecitabine, cisplatin, gemcitabine, and epirubicin.

  In one embodiment of the invention, the anti-CTLA4 and aromatase inhibitor combination is administered in further combination with paclitaxel and an additional agent selected from the group consisting of carboplatin, cisplatin, and gemcitabine. In one aspect, the additional agent is carboplatin.

  In another embodiment, the combination of anti-CTLA4 and an aromatase inhibitor is administered with an additional agent, and the additional agent is a vinca alkaloid. In one aspect, the vinca alkaloid is vinorelbine.

  In one embodiment of the invention, the additional agent administered with the antibody-aromatase inhibitor combination is a camptothecin analog. In one aspect, the camptothecin analog is irinotecan.

  In another embodiment, the additional agent is an EGFR inhibitor.

  In one embodiment of the invention, the additional agent is an erbB2 inhibitor. In one aspect, the erbB2 inhibitor is selected from trastuzumab and pertuzumab. In yet another aspect, the erbB2 inhibitor is trastuzumab (HERCEPTIN).

  In one embodiment of the invention, the anti-CTLA4 and aromatase inhibitor combination is administered in further combination with a combination comprising 5-FU, doxorubicin, and cyclophosphamide.

  In another embodiment of the invention, the anti-CTLA4 and aromatase inhibitor combination is administered in further combination with a combination comprising docetaxel, doxorubicin, and cyclophosphamide.

  The methods of the present invention also relate to cancer treatment in mammals, preferably humans, that have undergone stem cell transplantation, wherein the method comprises treating a mammal with an effective amount of human anti-cancer in combination with an aromatase inhibitor. Administering CTLA4 antibody in further combination with stem cell transplantation. The stem cell transplant may be an allogeneic or autologous stem cell transplant, and the aromatase inhibitor is more preferably exemestane. If the method involves stem cell transplantation, the initial amount of the antibody-aromatase inhibitor combination can be administered after the mammal's immune system has recovered from transplantation, for example, 1-12 months after transplantation. In certain embodiments, the initial dose is administered 1 to 3 months, or 1 to 4 months after transplantation. Patients can receive preparatory treatment for stem cell transplantation.

VI. Pharmaceutical composition The present invention comprises the formulation and use of a pharmaceutical composition comprising as an active ingredient the human anti-CTLA4 antibody of the present invention in combination with an aromatase inhibitor, preferably a steroidal aromatase inhibitor, wherein the aromatase inhibitor is exemestane Even more preferably. Such a pharmaceutical composition is in a form suitable for administration to a subject comprising only each active ingredient as a combination of at least one active ingredient (eg, an effective amount of anti-CTLA4, an effective amount of an aromatase inhibitor). Or a pharmaceutical composition may comprise an active ingredient and one or more pharmaceutically acceptable carriers, one or more additional (active and / or inactive) ingredients, or some combination thereof. It may be included.

  In one embodiment, the antibody is administered parenterally (eg, intravenously) in an aqueous solution and the aromatase inhibitor (eg, exemestane) is administered orally in the form of a pill / capsule. However, one of ordinary skill in the art will know based on the disclosure provided herein that the invention is not limited to these or any other formulation, dose, route of administration, and the like. Rather, the present invention includes, but is not limited to, among many others, each drug in a different formulation and administered separately by different routes of administration, and the antibody and aromatase inhibitor in a single composition (e.g., Any formulation or method in which the antibody is administered in combination with an aromatase inhibitor, including those administered as, in particular, intravenously administered aqueous compositions. Thus, the following discussion describes various formulations for practicing the methods of the invention involving the administration of any anti-CTLA4 antibody in combination with any aromatase inhibitor, but the invention is limited to these formulations. Rather, any formulation that can be readily determined by one of ordinary skill in the art having the teachings provided herein for use in the methods of the present invention is included.

  The antibody used in the present invention can be incorporated into a pharmaceutical composition suitable for administration to a subject. Usually, the pharmaceutical composition comprises the antibody and a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable carrier” includes any physiologically compatible solvent, dispersion medium, skin, antibacterial and antifungal agent, isotonic absorption delaying agent, and the like. . Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerin, ethanol, and the like, as well as combinations thereof. In many cases, it will be preferable to include isotonic substances, for example, sugars, polyalcohols such as mannitol or sorbitol, or sodium chloride in the composition. Pharmaceutically acceptable substances such as wetting or emulsifying agents, preservatives, or small amounts of auxiliary substances such as buffers improve the lifetime or effectiveness of the antibody or antibody portion.

  The antibody may be in various forms. These forms include, for example, liquid, semi-solid, such as liquid solutions (eg, injection and infusion solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. , And solid dosage forms. The preferred form varies depending on the intended mode of administration and therapeutic application. A typical preferred composition is in the form of a solution for injection or infusion, such as a composition similar to that used for passive human immunization with other antibodies. The preferred mode of administration is parenteral (eg, intravenous, subcutaneous, intraperitoneal, intramuscular). In preferred embodiments, the antibody is administered by intravenous infusion or injection. In another preferred embodiment, the antibody is administered by intramuscular or subcutaneous injection.

  A therapeutic composition usually must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable to high drug concentration. Sterile injectable solutions can be prepared by incorporating the required amount of antibody into a suitable solvent, optionally with one or a combination of the components listed above, followed by filtration and sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for formulating sterile injectable solutions, the preferred formulation methods are vacuum drying and lyophilization resulting in a powder of the active ingredient and any additional desired ingredients from the previously sterile filtered solution. The proper fluidity of the solution can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Absorption of injectable compositions can be extended by including in the composition an agent that delays absorption, for example, monostearate salts or gelatin.

  The antibodies can be administered by a variety of methods known in the art including, but not limited to, oral, parenteral, mucosal, inhalation, topical, buccal, nasal, and rectal. For many therapeutic applications, the preferred route / mode of administration is subcutaneous, intramuscular, intravenous, or infusion. If desired, needle-free injection can be used. As will be appreciated by those skilled in the art, the route and / or mode of administration will vary depending on the desired outcome.

  In certain embodiments, antibodies may be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. it can. Biodegradable biocompatible polymers can be used such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid and the like. Many methods for preparing such formulations are patented or generally known to those skilled in the art. For example, “Sustained and Controlled Release Drug Delivery Systems”, J. Am. R. Edited by Robinson, Marcel Dekker, Inc. See, New York, USA (1978).

  Dosage regimens can be adjusted to provide the optimum desired response. For example, a single bolus administration may be performed, administration may be divided into several times over time, or the dose may be increased or decreased depending on the urgency of the treatment situation. It is particularly advantageous to formulate parenteral compositions in dosage unit form because they are easy to administer and make the dosage uniform. Dosage unit form, as used herein, refers to a physically discrete unit suitable as a unit dosage for a mammalian subject undergoing treatment, each unit being combined with the required pharmaceutical carrier as desired. Contains a predetermined amount of active compound calculated to produce a therapeutic effect. The dosage unit form specifications of the present invention include (a) the unique properties of an antibody, as well as the specific therapeutic or prophylactic effect to be realized, and (b) an active compound for treating individual susceptibility. It is inevitably determined by the limitations inherent in the industry to formulate and varies directly accordingly.

  It should be noted that dosage values may vary depending on the type and severity of the condition to be alleviated and may include one or more doses. The detailed dosing schedule should be adjusted over time for any particular subject according to individual needs and the professional judgment of the person administering the composition, and as described herein. It should be further understood that the dosage ranges given are exemplary only and do not limit the scope or operation of the claimed compositions.

  In one embodiment, the antibody comprises sodium acetate, polysorbate 80, and sodium chloride with 5 mg / ml, more preferably about 10 mg / ml, even more preferably about 15 mg / ml, and even more preferably about 20 mg / ml of antibody. It is administered as an intravenous formulation as a sterile aqueous solution containing a pH in the range of about 5-6. The intravenous formulation is preferably a sterile aqueous solution at pH 5.5 containing 20 mM sodium acetate, 0.2 mg / ml polysorbate 80, and 140 mM sodium chloride with 5 or 10 mg / ml antibody. In addition, solutions containing anti-CTLA4 antibodies are known in histidine, mannitol, sucrose, trehalose, glycine, poly (ethylene) glycol, EDTA, methionine, and any combination thereof, and related industries, among many other compounds. Many other compounds that have been identified may be included.

  In one embodiment, a portion of the antibody formulation dose is administered by intravenous bolus administration and the remainder is administered by infusion. For example, an intravenous injection of 0.01 mg / kg antibody can be given as a bolus dose and the remainder of a given antibody dose can be administered by intravenous injection. The predetermined dose of antibody can be administered over a period of, for example, 1.5 hours to 2 hours to 5 hours.

  With respect to an aromatase inhibitor, the inhibitor is a pharmaceutical in the form of a physiologically acceptable ester or salt, such as in combination with a physiologically acceptable cation or anion, as is well known in the art. It can be present in the composition.

  Formulations of the pharmaceutical compositions described herein may be prepared by any method known or later developed in the field of pharmacokinetics. In general, such preparative methods involve combining the active ingredient with a carrier or one or more other accessory ingredients and then, if necessary or desired, shaping the product into the desired unit or units. Packaging.

  Pharmaceutical compositions useful in the methods of the invention are prepared, packaged, or sold in oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, ocular, or other suitable route of administration. can do. Other contemplated formulations include contemplated nanoparticles, liposomal formulations, resealed erythrocytes containing the active ingredient, and immunologically based formulations.

  The pharmaceutical compositions of the invention can be prepared, packaged, or sold in large quantities as a single unit dose or multiple single unit doses. As used herein, a “unit dose” is an individual amount of a pharmaceutical composition that contains a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient to be administered to the subject, or is expedient for that dosage, eg one half or one third of such dosage It is a fraction.

  The relative amounts of the active ingredient, pharmaceutically acceptable carrier, and any additional ingredients in the pharmaceutical compositions of the present invention may further depend on the individuality, size, and condition of the subject being treated. It depends on the route of administration. By way of example, the composition may comprise between 0.1% and 100% (w / w) active ingredient.

  In addition to the active ingredient, the pharmaceutical composition of the present invention may further comprise one or more additional pharmaceutically active agents. Additional substances specifically contemplated include antiemetics, antidiarrheal drugs, chemotherapeutic agents, cytokines and the like.

  Controlled or sustained release formulations of the pharmaceutical compositions of the invention can be manufactured using conventional techniques.

  Formulations of the pharmaceutical composition of the present invention suitable for oral administration include, but are not limited to, individual solids, including tablets, hard or soft capsules, cachets, troches, or lozenges each containing a predetermined amount of the active ingredient. It can be prepared, packaged, or sold in dosage unit form. Other formulations suitable for oral administration include, but are not limited to, powder or granular formulations, aqueous or oily suspensions, aqueous or oily solutions, or emulsions.

  As used herein, an “oily” liquid includes liquid molecules that contain carbon and has a polar character that is weaker than water.

  A tablet containing the active ingredient can be prepared, for example, by compressing or molding the active ingredient, optionally with one or more additional ingredients. Compressed tablets are in a free-flowing form, such as a powder or granular formulation, optionally mixed with one or more of binders, lubricants, pharmaceutical additives, surfactants, and dispersants. The active ingredient can be prepared by compression in a suitable device. Molded tablets may be made by molding in a suitable apparatus a mixture consisting of the active ingredient, a pharmaceutically acceptable carrier, and at least a liquid sufficient to wet the mixture.

  Pharmaceutically acceptable excipients used in the manufacture of tablets include, but are not limited to, inert diluents, granulating and disintegrating agents, binders, and lubricants. Known dispersants include, but are not limited to, potato starch and sodium starch glycolate. Known surfactants include, but are not limited to, sodium lauryl sulfate. Known diluents include, but are not limited to, calcium carbonate, sodium carbonate, lactose, microcrystalline cellulose, calcium phosphate, calcium hydrogen phosphate, and sodium phosphate. Known granulating disintegrants include, but are not limited to, corn starch and alginic acid. Known binders include, but are not limited to, gelatin, acacia, pregelatinized corn starch, povidone, and hydroxypropyl methylcellulose. Known lubricants include, but are not limited to, magnesium stearate, stearic acid, silica, and talc.

  The tablets may be uncoated or coated using known methods to achieve slow degradation in the subject's gastrointestinal tract, thereby providing sustained release and absorption of the active ingredient. May be. By way of example, materials such as glyceryl monostearate and glyceryl distearate can be used to coat tablets. Further, by way of example, coating tablets using the methods described in US Pat. Nos. 4,256,108, 4,160,452, and 4,265,874 can produce osmotic slow-release tablets. The tablets may further comprise sweetening agents, flavoring agents, coloring agents, preservatives, or some combination thereof in order to provide a pharmaceutically refined and palatable formulation.

  Hard capsules containing the active ingredient can be manufactured using a physiologically degradable composition such as gelatin. Such hard capsules contain the active ingredient and may further contain additional ingredients including, for example, an inert solid diluent such as calcium carbonate, calcium phosphate, kaolin.

  Soft gelatin capsules containing the active ingredient can be manufactured using a physiologically degradable composition such as gelatin. Such soft capsules contain the active ingredients which may be mixed with water or an oil medium such as peanut oil, liquid paraffin, olive oil.

  Liquid formulations of the pharmaceutical composition of the present invention suitable for oral administration are prepared, packaged, and sold in liquid form or in the form of a dry product that is to be reconstituted with water or another suitable excipient prior to use. can do.

  Suspensions may be prepared using conventional methods to obtain an aqueous or oily excipient suspension of the active ingredient. Aqueous excipients include, for example, water and isotonic saline. Oily excipients include, for example, tonsil oil, oily esters, ethyl alcohol; vegetable oils such as peanut, olive, sesame, coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. Suspensions include, but are not limited to, one or more including suspending agents, dispersing or wetting agents, emulsifying agents, demulcents, preservatives, buffers, salts, flavoring agents, coloring agents, and sweetening agents. Additional components may also be included. The oily suspension may further comprise a thickener. Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fat, sodium alginate, povidone, tragacanth gum, acacia gum, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, etc. Is included. Known dispersing or wetting agents include, but are not limited to, naturally occurring phosphatides such as lecithin; alkylene oxides and partial esters derived from fatty acids, long chain fatty alcohols, fatty acids and hexitol, or fatty acids and Condensates with partial esters obtained from anhydrous hexitol (for example, polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, polyoxyethylene sorbitan monooleate) are included. Known emulsifiers include, but are not limited to, lecithin and acacia. Known preservatives include, but are not limited to, methyl, ethyl, or n-propyl p-hydroxybenzoate, ascorbic acid, and sorbic acid. Known sweetening agents include, for example, glycerin, propylene glycol, sorbitol, sucrose, and saccharin. Known thickening agents for oily suspensions include, for example, beeswax, hard paraffin, and cetyl alcohol.

  Liquid solutions of the active ingredient in aqueous or oily solvents can be prepared in substantially the same manner as suspensions, the main difference being that the active ingredient is dissolved rather than suspended in the solvent. Liquid solutions of the pharmaceutical composition of the present invention include each of the components described with respect to the suspension, but it is understood that the suspending agent does not necessarily aid in dissolving the active ingredient in the solvent. Aqueous solvents include, for example, water and isotonic saline. Oily solvents include, for example, tonsil oil, oily esters, ethyl alcohol; vegetable oils such as peanut, olive, sesame or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin.

  Powder and granule formulations of the pharmaceutical formulation of the present invention can be prepared using known methods. Such formulations may be administered directly to the subject, for example, producing tablets, filling capsules, or adding aqueous or oily excipients thereto to form aqueous or oily suspensions or solutions. This may be used to prepare. Each of these formulations may further comprise one or more of a dispersing or wetting agent, a suspending agent, and a preservative. Additional pharmaceutical additives such as fillers, sweetening agents, flavoring agents, coloring agents and the like can also be included in these formulations.

  The pharmaceutical composition of the present invention can also be prepared, packaged or sold in the form of an oil-in-water emulsion or a water-in-oil emulsion. The oily phase may be a vegetable oil such as olive or arachis oil, a mineral oil such as liquid paraffin, or a combination of these. Such compositions are obtained from combinations of fatty acids and hexitol anhydrides such as naturally occurring gums such as gum acacia and tragacanth, naturally occurring phosphatides such as soybean or lecithin phosphatide, sorbitan monooleate, etc. One or more emulsifiers may further be included such as esters or partial esters, as well as condensates of such partial esters and ethylene oxide, such as polyoxyethylene sorbitan monooleate. These emulsions may contain additional ingredients including, for example, sweetening or flavoring agents.

  The pharmaceutical composition of the present invention can also be prepared, packaged or sold in a formulation suitable for rectal administration. Such a composition may be in the form of, for example, a suppository, a retention enema preparation, a solution for rectal or colonic irrigation.

  Suppository formulations are pharmaceutically acceptable non-irritating that are solid with the active ingredient and solid at normal room temperature (ie about 20 ° C.) and liquid at the subject's rectal temperature (ie about 37 ° C. in healthy subjects). It can be produced by mixing with excipients. Suitable pharmaceutically acceptable excipients include, but are not limited to, cocoa butter, polyethylene glycol, and various glycerides. Suppository formulations may further comprise various additional ingredients including, but not limited to, antioxidants and preservatives.

  Retentive enema preparations or solutions for rectal or colonic irrigation can be prepared by mixing the active ingredient with a pharmaceutically acceptable liquid carrier. As is well known in the art, enema formulations can be administered and packaged within a delivery device adapted to the subject's rectal anatomy. Enema formulations may further include various additional ingredients including, but not limited to, antioxidants and preservatives.

  The pharmaceutical composition of the present invention can also be prepared, packaged or sold in a formulation suitable for vaginal administration. Such a composition may be in the form of, for example, a suppository; an impregnated or coated vaginal material such as a tampon, a pressurized formulation, etc .; a gel or cream; or a solution for vaginal irrigation.

  Methods for impregnating a material with a chemical composition or coating a material with a chemical composition are known in the art, including, but not limited to, methods for attaching or bonding a chemical composition to a surface, and synthesizing a material. Methods of incorporating a chemical composition into the structure of the material (ie, with a physiologically degraded material), as well as absorbing an aqueous or oily solution or suspension into the absorbent material, followed by drying or drying Includes methods that do not.

  Injectable formulations or vaginal lavage solutions can be prepared by mixing the active ingredient with a pharmaceutically acceptable liquid carrier. As is well known in the art, infusion formulations can be administered and packaged within a delivery device adapted to the subject's vaginal anatomy. Injectable formulations may further comprise various additional ingredients including, but not limited to, antioxidants, antibiotics, antifungal agents, and preservatives.

  As used herein, “parenteral administration” of a pharmaceutical composition includes any route of administration characterized by physical breakthrough of the tissue of interest and administration of the pharmaceutical composition through that breakthrough of tissue. Thus, for parenteral administration, administration of the pharmaceutical composition includes, but is not limited to, injection of the composition, application of the composition through a surgical incision, application of the composition through a non-surgical wound that penetrates tissue, etc. Is included. In particular, parenteral administration is believed to include, but is not limited to, subcutaneous, intraperitoneal, intramuscular, intrasternal injection, and renal dialysis infusion techniques.

  Formulations of pharmaceutical compositions suitable for parenteral administration comprise the active ingredient in combination with a pharmaceutically acceptable carrier such as sterile water or sterile isotonic saline. Such formulations can be prepared, packaged, or sold in a form suitable for bolus administration or continuous administration. Formulations for injection can be prepared, packaged, or sold in unit dosage forms such as ampoules or multi-dose containers containing preservatives. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous excipients; pastes; and implantable sustained or biodegradable as discussed below. Sex preparations are included. Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending, stabilizing, or dispersing agents. In one embodiment of a formulation for parenteral administration, the active ingredient is reconstituted with a suitable excipient (eg, sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition. In dry (ie, powder or granule) form.

  The compositions of the present invention can be administered by a variety of methods known in the art. The route and / or mode of administration will vary depending on the desired outcome. Active compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid can be used. Many methods for preparing such formulations are described, for example, in “Sustained and Controlled Release Drug Delivery Systems”, J. Org. R. Edited by Robinson, Marcel Dekker, Inc. , New York (1978). The pharmaceutical composition is preferably produced under GMP conditions.

  The pharmaceutical compositions can be prepared, packaged, or sold in the form of a sterile aqueous or oily suspension or solution for injection. This suspension or solution may be formulated according to known techniques and may contain, in addition to the active ingredient, additional ingredients such as dispersants, wetting agents, suspending agents described herein. Such sterile injectable formulations can be prepared using parenterally acceptable non-toxic diluents or solvents such as water or 1,3-butanediol. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or diglycerides. Other useful formulations that can be administered parenterally include those that contain the active ingredient in microcrystalline form, in a liposomal formulation, or as a component of a biodegradable polymer system. Sustained release or implantable compositions may include pharmaceutically acceptable polymeric or hydrophobic materials such as emulsions, ion exchange resins, slightly soluble polymers, slightly soluble salts.

  Formulations suitable for topical administration include, but are not limited to, liniments, lotions; oil-in-water or water-in-oil emulsions such as creams; ointments or pastes; liquid or semi-liquid preparations such as solutions or suspensions. It is. Formulations for topical administration contain, for example, from about 1% to about 10% (w / w) active ingredient, but the concentration of the active ingredient may be as high as the limit at which the active ingredient is soluble in the solvent. Formulations for topical administration may further comprise one or more of the additional ingredients described herein.

  The pharmaceutical composition of the present invention can be prepared, packaged, or sold in a formulation suitable for administration to the lungs from the buccal cavity. Such formulations may comprise dry particles comprising the active ingredient and having a diameter in the range of about 0.5 to about 7 nanometers, about 1 to about 6 nanometers. Such compositions can be administered using a device that includes a dry powder reservoir in which a propellant stream is directed to disperse the powder, or dissolved in a low boiling propellant in a closed container. Conveniently in the form of a dry powder for administration using a self-propelling solvent / powder dispensing container, such as a device containing the suspended active ingredient. Such a powder preferably consists of particles in which at least 98% by weight of the particles have a diameter greater than 0.5 nanometers and at least 95% by weight of the particles have a diameter of less than 7 nanometers. More preferably, the diameter of at least 95% by weight of the particles is greater than 1 nanometer and the diameter of at least 90% by volume of the particles is less than 6 nanometers. The dry powder composition preferably comprises a solid fine powder diluent such as sugar and is conveniently provided in unit dosage form.

  Low boiling propellants generally include liquid propellants having a boiling point below 65 ° F. at atmospheric pressure. Generally, the propellant may account for 50 to 99.9% (w / w) of the composition and the active ingredient may account for 0.1 to 20% (w / w) of the composition. The propellant may further comprise additional components such as a liquid nonionic surfactant, a solid anionic surfactant, or a solid diluent (preferably having the same particle size as the particles containing the active component).

  A pharmaceutical composition of the invention formulated for pulmonary delivery may provide the active ingredient in the form of droplets of a solution or suspension. Such formulations can be prepared, packaged, or sold as an aqueous or dilute alcoholic solution or suspension, which may be sterile, containing the active ingredient, using any spraying or atomizing device. It can be conveniently administered. Such formulations may contain one or more additional ingredients including but not limited to flavoring agents such as sodium saccharin, volatile oils, buffers, surfactants, or preservatives such as methyl hydroxybenzoate. Further may be included. The droplets provided by this route of administration preferably have an average diameter in the range of about 0.1 to about 200 nanometers.

  The formulations described herein useful for pulmonary delivery are also useful for intranasal delivery of the pharmaceutical compositions of the invention.

  Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle from about 0.2 to 500 micrometers. Such formulations are administered in a sniffing manner, i.e. by rapid inhalation through the nose from a powder container held near the nostril.

  Formulations suitable for nasal administration may contain, for example, only about 0.1% (w / w), a large amount of 100% (w / w) active ingredient, one of the additional ingredients described herein. Species or multiples may further be included.

  The pharmaceutical composition of the present invention can be prepared, packaged or sold in a formulation suitable for buccal administration. Such formulations can be, for example, in the form of tablets or lozenges manufactured using conventional methods, for example, compositions that are orally dissolved or degraded, and optional additions described herein. The active ingredient may be 0.1 to 20% (w / w) containing one or more of the ingredients. Alternatively, formulations suitable for buccal administration may comprise a powder containing the active ingredient or an aerosolized or nebulized solution or suspension. Such powdered or aerosolized formulations preferably have an average particle size or droplet size in the range of about 0.1 to about 200 nanometers when dispersed, as described herein. One or more of the components may further be included.

  The pharmaceutical composition of the present invention can be prepared, packaged or sold in a formulation suitable for administration to the eye. Such formulations may, for example, be in the form of eye drops comprising a 0.1-1.0% (w / w) solution or suspension of the active ingredient in an aqueous or oily liquid carrier, for example. Such droplets may further include buffering agents, salts, or one or more other of the additional components described herein. Other useful formulations that can be administered to the eye include those that contain the active ingredients of a microcrystalline form or a liposomal formulation.

  As used herein, “additional ingredients” include, but are not limited to, the following: pharmaceutical additives; surfactants; dispersants; inert diluents; granulating disintegrants; Sweeteners; flavoring agents; coloring agents; preservatives; physiologically degraded compositions such as gelatin; aqueous excipients and solvents; oily excipients and solvents; suspending agents; Emulsifiers, demulcents; buffers; salts; thickeners; fillers; emulsifiers; antioxidants; antibiotics; antifungal agents; stabilizers; and pharmaceutically acceptable polymers or hydrophobic materials Species or plurals are included. Other “additional ingredients” that can be included in the pharmaceutical compositions of the present invention are known in the art, eg, “Remington's Pharmaceutical Sciences”, edited by Genaro, incorporated herein by reference. Mack Publishing Co. , Easton, Pennsylvania, USA (1985).

  The anti-CTLA4 / aromatase inhibitor active ingredient combination of the present invention can be administered to animals, preferably humans. The exact dose administered for each active ingredient will vary depending on several factors including, but not limited to, the type of animal being treated and the type of disease state, the age of the animal, and the route of administration. Become.

  The anti-CTLA4 antibody may be administered to the animal several times daily, or less frequently, such as once a day, once a week, once every two weeks, once a month, or once every few months. Furthermore, it may be administered at a smaller number of times such as once or less per year. The number of doses will be readily apparent to those skilled in the art and will depend on several factors such as, but not limited to, the type and severity of the disease being treated, the type of animal and the age.

  The aromatase inhibitor, preferably exemestane, may be administered to the animal several times daily, or less frequently, such as once a day, once a week, once every two weeks, once a month, or It may be administered less frequently, such as once every few months or even once a year or less. The number of doses will be readily apparent to those skilled in the art and will depend on several factors such as, but not limited to, the type and severity of the disease being treated, the type of animal and the age.

  The antibody and aromatase can be co-administered separately, on different dates, or at different times of the day, or at the same time or on the same day. Thus, co-administration is any combination of antibody and aromatase inhibitor administration in which administration of the two drugs mediates a detectably greater patient benefit than administration of either drug in the absence of the other. Includes temporal combinations.

  The antibody-aromatase inhibitor combinations of the present invention can be co-administered with numerous other compounds (among many others, other anti-hormonal therapeutics, cytokines, chemotherapeutics and / or antivirals). it can. Alternatively, the compounds can be administered 1 hour, 1 day, 1 week, 1 month or more prior to the antibody-aromatase inhibitor combination or any combination thereof. Further, the compound is administered 1 hour, 1 day, 1 week or more after administration of radiation, stem cell transplant, or any therapeutic agent (eg, cytokine, chemotherapeutic compound, etc.) or any combination thereof. be able to. The frequency and dosage regimen will be readily apparent to those skilled in the art and include, but are not limited to, the type and severity of the disease being treated, the age and health of the animal, the identity of the compound, or the compound being administered, It depends on several factors such as the route of administration of the various compounds.

VII. Kits The present invention includes a variety of kits comprising applicators and therapeutic materials that describe the use of a combination to perform a method of the invention with a therapeutically effective amount of a human anti-CTLA4 antibody of the invention and a therapeutically effective amount of an aromatase inhibitor. including. Exemplary kits are described below, but other useful kit contents will be apparent to those skilled in the art in light of this disclosure. Each of these kits is within the scope of the present invention.

  The present invention includes a kit for treating breast cancer in a patient in need thereof. The kit includes a human anti-CTLA4 antibody of the present invention and at least one aromatase inhibitor. This inhibitor includes, but is not limited to, fulvestrant, anastrozole, letrozole, and exemestane. The kit further includes, but is not limited to, an applicator that includes a syringe for administering the components of the kit to the patient. In addition, the kit includes explanatory material that describes information relating to the use of the kit to treat breast cancer in a patient.

  The kit consists of antibodies 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 11.6. 1, 11.7.1. At least one anti-CTLA4 antibody selected from antibodies having heavy and light chain amino acid sequences of 12.3.1.1, 12.9.1.1, and 10D1 (MDX-010) More preferably, the antibody is even more preferably an antibody having the heavy and light chain amino acid sequences of antibodies 4.13.1, 11.2.1, and 10D1 (MDX-010).

  In one embodiment, the aromatase inhibitor is at least one inhibitor selected from anastrozole, letrozole, and exemestane. More preferably, the aromatase inhibitor is exemestane.

  The kit may include a number of additional agents for the treatment of breast cancer. Such agents are those previously described, and among many others, include chemotherapeutic compounds, cancer vaccines, signal transduction inhibitors, agents useful in the treatment of abnormal cell proliferation or cancer, IGF-1R. This includes antibodies or other ligands that bind and inhibit tumor growth, as well as cytokines.

  The invention also provides a product (eg, intravenous) containing an effective amount (eg, greater than 10 mg / kg, at least 15 mg / kg, or 15 mg / kg) of a human anti-CTLA4 antibody and a therapeutically effective amount of an aromatase inhibitor. Dosage form adapted for internal administration). In certain embodiments, the product comprises one or more containers comprising a human anti-CTLA4 antibody, an aromatase inhibitor, and a label and / or instructions for use for cancer treatment.

  The present invention will be described in more detail with reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not limiting unless otherwise specified. Accordingly, the present invention should in no way be construed as limited to the following examples, but rather should be construed to include any variations that become apparent as a result of the teaching provided herein.

Anti-CLTA4 antibody in combination with exemestane in metastatic or locally advanced breast cancer in post-menopausal patients At least one lesion can be accurately measured in two dimensions, and its size is ≧ 2 cm × by conventional CT scan Patients with metastatic or locally advanced breast cancer that was 1 cm, or ≧ 1 cm × 1 cm on a helical CT scan, received IV infusion (100 mL / hr) of anti-CTLA4 antibody as described herein. Give prophylactic antiemetics and antidiarrheal drugs as needed.

  The treatment is repeated after 28 days. Tolerable toxicities or diseases that are not tolerated for up to 12 cycles, starting at 3 mg / kg over a cohort over time and then increasing to 6 mg / kg, 10 mg / kg, and 15 mg / kg each 28 days The dose of anti-CTLA4 antibody is given incrementally without progression.

  Preferably, the patient is premedicated with an antihistamine (H1) at least one and a half hours prior to infusion of anti-CTLA4. Premedication is recommended but not required.

  All patients receive exemestane as a continuous oral dose of 25 mg per day for a total 28-day cycle. Exemestane is preferably taken at the same time every day.

  The dose is increased in stages using an accelerated titration design utilizing a dose-doubling scheme with 3-6 subjects per cohort. Within each new cohort, no waiting period between subjects is required. A subsequent cohort cannot be opened until the first subject at the current dose level is observed for 21 days and the subsequent subject is observed for 14 days.

  The antibody is provided in a 10 ml clear glass vial with a rubber stopper and an aluminum seal. Each vial is 5 mg / ml (nominally 50 mg / vial) anti-CTLA4 antibody in a sterile aqueous solution of pH 5.5 containing 20 mM sodium acetate, 0.2 mg / ml polysorbate 80, and 140 mM sodium chloride. Containing.

  All patients are assessed for ECOG performance status, vital signs, and body weight prior to administration, and vital signs can be repeated after administration, as clinically indicated. A physical examination (including ophthalmological assessment and autoimmune signature) is performed on day 1. Hematology panel (hematocrit, RBC count, WBC count, difference), chemistry (alkaline phosphatase, calcium, chloride, GGT, LDH, magnesium, phosphorus, random glucose, sodium, urea, uric acid), urinalysis (blood, Protein), others (activated partial thromboplastin time [APTT], prothrombin time (PT), autoantibody panel, C-reactive protein, TSH, T3, T4, amylase, lipase, serum C3, C4, serum Ig levels) Get a sample of.

  Baseline human anti-human antibody (HAHA) titers are determined and pharmacokinetic (PK) specimens are obtained prior to administration.

  The following endpoints are measured: PK parameters, HAHA, response rate, and growth inhibition time. The Kaplan-Meier aggressive limit method is used to calculate growth inhibition time and overall survival time.

  The anti-CTLA4 antibody preferably has the heavy and light chain amino acid sequences of at least one antibody selected from 4.1.1, 4.13.1, 11.2.1, and 10D1. More preferably, the antibody has a heavy and light chain amino acid sequence of 11.2.1.

  Three patients received 3 mg / kg of anti-CTLA4 antibody (ticilimumab). All three patients received 25 mg of exemestane daily. One patient received anti-CTLA4 twice every 28 days and exemestane daily for 56 days. Cycle 2 staging CT revealed no lung lesions at baseline. Patients also showed new mild peripheral wall thickening, including unknown etiology, possible infections, or inflamed terminal ileum, distal descending colon, and sigmoid colon. The patient had no medical history explaining these changes, felt healthy and was clinically asymptomatic. No infection was detected. This patient declined to investigate disease progression (PD).

  The second patient received two cycles of anti-CTLA4 antibody and received approximately 14 days of exemestane therapy (25 mg daily). After the first ticilimumab administration, the patient reported that some baseline lymphedema of the right arm was somewhat worse and then sedated. The patient also reported a transient mild non-pruritic rash.

  The third patient has so far received one cycle of ticilimumab and 25 mg of exemestane daily for about 28 days. On day 28, routine laboratory investigations detected ALP and AST elevated to stage 1 and ALT elevated to stage 2. The patient was asymptomatic. The therapy was put on hold with the improvement of these test values unresolved.

  These data suggest that patients treated with a combination of ticilimumab and exemestane showed biological activity.

  The disclosures of any patents, patent applications, and publications cited herein are hereby incorporated herein by reference in their entirety.

  Although the invention has been disclosed with reference to detailed embodiments, it is to be understood that other embodiments and variations of the invention may be devised by others skilled in the art without departing from the spirit and scope of the invention. Needless to say. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

1A-1D included. It is a sequence table which shows the nucleotide sequence and amino acid sequence of anti-CTLA4 antibody 4.1.1. FIG. 1A shows the full length nucleotide sequence (SEQ ID NO: 1) of the 4.1.1 heavy chain. FIG. 1B shows the 4.1.1 heavy chain full-length amino acid sequence (SEQ ID NO: 2) and the 4.1.1 heavy chain variable region amino acid sequence (SEQ ID NO: 3) shown between brackets “[]”. Show. The amino acid sequence of each 4.1.1 heavy chain CDR is underlined. The CDR sequences are as follows: CDR1 is GFTFSHGHMH (SEQ ID NO: 4), CDR2 is VIWYDGRNKYYADSV (SEQ ID NO: 5), and CDR3 is GHHFGPFDY (SEQ ID NO: 6). FIG. 1C shows the nucleotide sequence of the 4.1.1 light chain (SEQ ID NO: 7). FIG. 1D shows the full length 4.1.1 light chain amino acid sequence (SEQ ID NO: 8) and the variable region (SEQ ID NO: 9) shown between brackets “[]”. The amino acid sequence of each CDR is shown as follows. That is, CDR1 is RASQSISSSFLA (SEQ ID NO: 10), CDR2 is GASSSRAT (SEQ ID NO: 11), and CDR3 is CQQYGTSPWT (SEQ ID NO: 12). Includes Figures 2A-2D. It is a sequence table which shows the nucleotide sequence and amino acid sequence of anti-CTLA4 antibody 4.13.1. FIG. 2A shows the full length nucleotide sequence of the 4.13.1 heavy chain (SEQ ID NO: 13). FIG. 2B shows the 4.13.1 heavy chain full-length amino acid sequence (SEQ ID NO: 14) and the 4.13.1 heavy chain variable region amino acid sequence (SEQ ID NO: 15) shown between brackets “[]”. Show. The amino acid sequence of each 4.13.1 heavy chain CDR is underlined. The CDR sequences are as follows: CDR1 is GFTFSHGIH (SEQ ID NO: 16), CDR2 is VIWYDGRNKDYADSV (SEQ ID NO: 12), and CDR3 is VAPLGPLDY (SEQ ID NO: 18). FIG. 2C shows the nucleotide sequence of the 4.13.1 light chain (SEQ ID NO: 19). FIG. 2D shows the amino acid sequence of the full length 4.13.1 light chain (SEQ ID NO: 20) and the variable region shown between brackets “[]” (SEQ ID NO: 21). The amino acid sequence of each CDR is shown as follows. That is, CDR1 is RASQSVSSYLA (SEQ ID NO: 22), CDR2 is GASSRAT (SEQ ID NO: 23), and CDR3 is CQQYGRSPFT (SEQ ID NO: 24). Includes Figures 3A-3D. It is a sequence table | surface which shows the nucleotide sequence and amino acid sequence of anti-CTLA4 antibody 11.2.1. FIG. 3A shows the full length nucleotide sequence of the 11.2.1 heavy chain (SEQ ID NO: 25). FIG. 3B shows the full length amino acid sequence of the 11.2.1 heavy chain (SEQ ID NO: 26) and the 11.2.1 heavy chain variable region amino acid sequence (SEQ ID NO: 27) shown between brackets “[]”. Show. The amino acid sequence of each 11.2.1 heavy chain CDR is underlined. The CDR sequences are as follows: CDR1 is GFTFSSYGMH (SEQ ID NO: 28), CDR2: VIWYDGSNKYYADSV (SEQ ID NO: 29), and CDR3: DPRGATYLYYYYGMMDV (SEQ ID NO: 30). FIG. 3C shows the nucleotide sequence of the 11.2.1 light chain (SEQ ID NO: 31). FIG. 3D shows the amino acid sequence of the full length 11.2.1 light chain (SEQ ID NO: 32) and the variable region (SEQ ID NO: 33) shown between brackets “[]”. The amino acid sequence of each CDR is shown as follows. That is, CDR1 is RASQSINSYLD (SEQ ID NO: 34), CDR2: AASSLQS (SEQ ID NO: 35), and CDR3: QQYYSTPFT (SEQ ID NO: 36).

Claims (20)

  A method of treating breast cancer in a patient in need of treatment of breast cancer, comprising administering to said patient a therapeutically effective amount of an anti-CTLA4 antibody or antigen-binding portion thereof in combination with a therapeutically effective amount of an aromatase inhibitor. .   The method of claim 1, wherein the aromatase inhibitor is at least one inhibitor selected from the group consisting of anastrozole, letrozole, and exemestane.   The method of claim 2, wherein the aromatase inhibitor is exemestane.   4. The method of claim 3, wherein the therapeutically effective amount of human anti-CTLA4 antibody amount is in the range of about 1 mg / kg to 40 mg / kg.   5. The method of claim 4, wherein the therapeutically effective amount of human anti-CTLA4 antibody amount is in the range of about 3 mg / kg to 15 mg / kg.   4. The method of claim 3, wherein the therapeutically effective amount of exemestane ranges from about 25 mg per day to 200 mg per day.   7. The method of claim 6, wherein the therapeutically effective amount of exemestane is about 25 mg per day.   The method of claim 1, wherein the treatment is selected from the group consisting of neoadjuvant chemotherapy, adjuvant therapy, first line therapy, second line therapy, and third line therapy.   2. The method of claim 1, wherein the antibody is selected from the group consisting of a non-human mammalian antibody, a chimeric antibody, and a human antibody.   The method of claim 9, wherein the antibody is a human antibody. Said anti-CTLA4 antibody or antigen-binding portion thereof is:
(A) a human antibody having a binding affinity for CTLA4 of about 10 ⁻⁸ or more and inhibiting the binding of CTLA4 and B7-1 and the binding of CTLA4 and B7-2;
(B) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 116.1 11.7.1. A human antibody having an amino acid sequence comprising at least one human CDR sequence corresponding to a CDR sequence from an antibody selected from the group consisting of: 12.3.1.1, 12.9.1.1, and 10D1.
(C) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 116.1 11.7.1. A human antibody having the heavy and light chain amino acid sequences of an antibody selected from the group consisting of: 12.3.1.1, and 12.9.1.1
(D) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 116.1 11.7.1. A human antibody having an amino acid sequence of a heavy chain variable region and a light chain variable region of an antibody selected from the group consisting of: 12.3.1.1, 12.9.1.1, and 10D1;
(E) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2.1, 116.1 11.7.1. , 12.3.1.1, 12.9.1.1, and 10D1 compete for binding of CTLA4 with at least one antibody having an antibody heavy and light chain amino acid sequence selected from the group consisting of An antibody or antigen-binding portion thereof, and (f) 4.1.1, 4.8.1, 4.10.2, 4.13.1, 4.14.3, 6.1.1, 11.2 .1, 11.6.1, 11.7.1. , 12.3.1.1, 12.9.1.1, and 10D1 cross-competing for the binding of CTLA4 with at least one antibody having heavy and light chain amino acid sequences of an antibody selected from the group consisting of The method according to claim 1, wherein the antibody is at least one antibody selected from the group consisting of an antibody or an antigen-binding portion thereof.   2. The method of claim 1, wherein the antibody is a human antibody having the heavy and light chain amino acid sequences of antibody 11.2.1. The antibody comprises a heavy chain and a light chain, and the amino acid sequences of the heavy chain variable region of the heavy chain and the light chain variable region of the light chain are as follows:
(A) the amino acid sequence of SEQ ID NO: 3 and the amino acid sequence of SEQ ID NO: 9,
(B) the amino acid sequence of SEQ ID NO: 15 and the amino acid sequence of SEQ ID NO: 21,
(C) the amino acid sequence of SEQ ID NO: 27 and the amino acid sequence of SEQ ID NO: 33,
(D) the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 1 and the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 7,
(E) the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 13 and the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 19,
(F) the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 25 and the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 31;
(G) The method according to claim 1, wherein the method is selected from the group consisting of the amino acid sequences of the heavy chain variable region and the light chain variable region of antibody 10D1. Said antibody or antigen-binding portion thereof is:
(A) having a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, and further comprising the amino acid sequences set forth in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12 An antibody having a light chain variable region,
(B) having a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, and further comprising the amino acid sequences set forth in SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24 An antibody having a light chain variable region,
(C) having a heavy chain variable region comprising the amino acid sequences set forth in SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, and further comprising the amino acid sequences set forth in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36 An antibody having a light chain variable region, and (d) a heavy chain variable region comprising the amino acid sequences of heavy chain CDR1, CDR2, and CDR3 of antibody 10D1, and further the amino acids of light chain CDR1, CDR2, and CDR3 of antibody 10D1 The method of claim 1, wherein the antibody is selected from the group consisting of antibodies having a light chain variable region comprising a sequence.   A tumor vaccine comprising the alkylating agent, folic acid antagonist, pyrimidine antagonist, anthracycline antibiotic, platinum compound, taxane, vinca alkaloid, camptothecin analog, toll-like receptor stimulant, heat shock protein Antigen-presenting cell-based therapy, mammalian rapamycin target protein inhibitor, erbB2 inhibitor, EGFR inhibitor, VEGF inhibitor, VEGFR inhibitor, angiogenesis inhibitor, antibody, immunomodulator, selective estrogen receptor modulator, 2. The method of claim 1, further comprising administering to the patient at least one agent selected from the group consisting of cytokines, tumor vaccines, antiproliferative drugs, immune co-stimulatory molecules, and cytokines. A therapeutically effective amount of an anti-CTLA4 antibody,
A pharmaceutical composition for treating breast cancer comprising a therapeutically effective amount of an aromatase inhibitor and a pharmaceutically acceptable carrier.   The pharmaceutical composition according to claim 16, wherein the aromatase inhibitor is at least one aromatase inhibitor selected from the group consisting of anastrozole, letrozole, and exemestane.   18. A pharmaceutical composition according to claim 17, wherein the aromatase inhibitor is exemestane.   Use of an amount of an anti-CTLA4 antibody in preparing a composition for the treatment of breast cancer in a patient, wherein the treatment further comprises administering an amount of an aromatase inhibitor to the patient.   20. The use according to claim 19, wherein the aromatase inhibitor is exemestane and the antibody is a human antibody having the heavy and light chain variable region amino acid sequences of antibody 11.2.1.

Images