Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
  • A61K38/179—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• methods of treating breast cancer having FGFR1 gene amplification, FGFR1 overexpression, FGFR3 gene amplification, FGFR3 overexpression, FGF2 gene amplification and/or FGF2 overexpression comprising
• the subject with breast cancer has previously been administered, or is currently being administered, trastuzumab (e.g., Herceptin®) and/or lapatinib (e.g., Tykerb®). In some embodiments, the subject has previously been trastuzumab (e.g., Herceptin®) and/or lapatinib (e.g., Tykerb®). In some embodiments, the subject has previously been trastuzumab (e.g., Herceptin®) and/or lapatinib (e.g., Tykerb®). In some embodiments, the subject has previously been administered, trastuzumab (e.g., Herceptin®) and/or lapatinib (e.g., Tykerb®). In some embodiments, the subject has previously been trastuzumab (e.g., Herceptin®) and/or lapatinib (e.g., Tykerb®). In some embodiments, the subject has previously been trastuzumab
• the aromatase inhibitor is selected from aminoglutethimide, testolactone (e.g., Teslac®), anastrozole (e.g., Arimidex®), letrozole (e.g., Femara®), exemestane (e.g., Aromasin®), vorozole (e.g., Rivisor®), formestane (e.g., Lentaron®), megestrol acetate (e.g., Megase®), and fadrozole (e.g., Afema®).
• testolactone e.g., Teslac®
• anastrozole e.g., Arimidex®
• letrozole e.g., Femara®
• exemestane e.g., Aromasin®
• vorozole e.g., Rivisor®
• formestane e.g., Lentaron®
• megestrol acetate e.g., Mega
• the subject has previously been administered, or is currently being administered, a GnRH antagonist.
• the GnRH agonist is selected from leuprolide (e.g., Lupron®, Eligard®), buserelin (e.g., Suprefact®, Suprecor®), histrelin (e.g., Supprelin LA®, Vantas®), goserelin acetate (e.g., Zoladex®), deslorelin (e.g., Suprelorin®, Ovuplant®), nafarelin (e.g., Synarel®), and triptorelin.
• leuprolide e.g., Lupron®, Eligard®
• buserelin e.g., Suprefact®, Suprecor®
• histrelin e.g., Supprelin LA®, Vantas®
• goserelin acetate e.g., Zoladex®
• deslorelin
• an AR inhibitor is selected from cyproterone acetate (e.g., Androcur®, Cyprostat®), flutamide (e.g., Eulexin®), bicalutamide (e.g., Casodex®), enzalutamide (e.g., Xtandi®), ketoconazole, and nilutamide (e.g., Anandron®, Nilandron®).
• a 17-hydroxylase inhibitor is abiraterone acetate (e.g., Zytiga®).
• FGFR1 fibroblast growth factor receptor 1
• ECD extracellular domain
• FGFR1 ECD fusion molecule a therapeutically effective amount of a fibroblast growth factor receptor 1 (FGFR1) extracellular domain (ECD) or an FGFR1 ECD fusion molecule.
• the subject has previously been administered, or is currently being administered, octreotide.
• therapeutically effective amount of octreotide has been previously administered, or is currently being administered to the subject.
• Nonlimiting exemplary ER antagonists include tamoxifen (e.g., Nolvadex®, Istabul®, Valodex®) and fulvestrant (e.g., Faslodex®).
• the ovarian cancer is estrogen receptor (ER) positive, progesterone (PR) positive, or ER positive and PR positive.
• a method comprises administering at least 5 mg/kg of an FGFRl ECD or an FGFRl ECD fusion molecule and at least 135 mg/m 2 paclitaxel and at least AUC 4 carboplatin to the subject.
• the method comprises administering from 135 mg/m 2 paclitaxel to 200 mg/m 2 paclitaxel, at least 175 mg/m 2 paclitaxel, from 175 mg/m 2 paclitaxel to 200 mg/m 2 paclitaxel, or 200 mg/m 2 paclitaxel.
• a method of treating lung cancer in a subject comprises administering at least 5 mg/kg of an FGFRl ECD or an FGFRl ECD fusion molecule and at least 40 mg/m 2 docetaxel. In some embodiments, the method comprises administering from 40 mg/m 2 docetaxel to 75 mg/m 2 docetaxel, at least 55 mg/m 2 docetaxel, from 55 mg/m 2 docetaxel to 75 mg/m 2 docetaxel, or 75 mg/m 2 docetaxel.
• the lung cancer is non-small cell lung cancer. In some embodiments, the non-small cell lung cancer is squamous non-small cell lung cancer.
• At least a portion of the cells of the cancer may overexpress at least one or two markers selected from DKK3 and FGF18. In any of the embodiments described herein, at least a portion of the cells of the cancer may overexpress ETV4. In some embodiments, the cancer does not have FGFRl gene amplification.
• the FGFRl ECD fusion molecule comprises a sequence selected from SEQ ID NO: 5 and SEQ ID NO: 6.
• the at least one fusion partner is an Fc and polyethylene glycol.
• the at least one fusion partners is polyethylene glycol.
• the fusion molecule comprises a linker between the FGFRl ECD and one or more fusion partners.
• the FGFRl ECD fusion molecule is FGFRl ECD.339-Fc.
• the therapeutically effective amount of the FGFRl ECD or FGFRl ECD fusion molecule is a dose of about 15 mg/kg body weight. In some embodiments, the therapeutically effective amount of the FGFRl ECD or FGFRl ECD fusion molecule is a dose of about 20 mg/kg body weight. In some embodiments, dosages may be administered twice a week, weekly, every other week, at a frequency between weekly and every other week, every three weeks, every four weeks, or every month.
• a method comprises determining whether at least a portion of the cancer cells in a sample obtained from the subject overexpress at least one, at least two, at least three, at least four, or at least five markers selected from FGFRl, FGFR3IIIc, FGF2, DKK3, FGF18, and ETV4, wherein overexpression is indicative of therapeutic
• FIG. 9 shows (A) FGFR1 and (B) FGFR3IIIc mR A expression in FGFR1-
• nucleic acid molecule and “polynucleotide” may be used interchangeably, and refer to a polymer of nucleotides. Such polymers of nucleotides may contain natural and/or non-natural nucleotides, and include, but are not limited to, DNA, RNA, and PNA.
• Nucleic acid sequence refers to the linear sequence of nucleotides that comprise the nucleic acid molecule or polynucleotide.
• FGFR1 extracellular domain (“FGFR1 ECD”) includes full-length FGFR1 ECDs, FGFR1 ECD fragments, and FGFR1 ECD variants.
• FGFR1 ECD refers to an FGFR1 polypeptide that lacks the intracellular and
• a human full-length FGFRl ECD may consist of the amino acid sequence corresponding to SEQ ID NO.: 2 (mature form) or to SEQ ID NO.: 1 (with the signal peptide).
• the term "FGFRl ECD fragment” refers to an FGFRl ECD having one or more residues deleted from the N and/or C terminus of the full- length ECD and that retains the ability to bind to FGF-2.
• the FGFRl ECD fragment may or may not include an N-terminal signal peptide.
• the FGFRl ECD fragment is a human FGFRl ECD fragment having an amino acid sequence corresponding to SEQ ID NO.: 4 (mature form) or to SEQ ID NO.: 3 (with the signal peptide).
• alterations of the reference sequence may occur at the N- or C- terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence, or in one or more contiguous groups within the reference sequence.
• whether any particular polypeptide is at least 70%, 80%, 90%, or 95% identical to, for instance, an amino acid sequence or to a polypeptide sequence encoded by a nucleic acid sequence set forth in the Sequence Listing can be determined conventionally using known computer programs, such the Bestfit program.
• hFGFRl-ECD.339 and “hFGFRl.339” may be used interchangeably to refer to the human FGFRl ECD corresponding to SEQ ID NO: 3 (with signal peptide) or to SEQ ID NO: 4 (without signal peptide; mature form).
• FGFRl ECD fusion molecule refers to a molecule comprising an FGFRl ECD, and one or more "fusion partners.”
• the FGFRl ECD and the fusion partner are covalently linked (“fused”).
• the fusion partner is also a polypeptide ("the fusion partner polypeptide")
• the FGFRl ECD and the fusion partner polypeptide may be part of a continuous amino acid sequence, and the fusion partner polypeptide may be linked to either the N terminus or the C terminus of the FGFRl ECD.
• the FGFRl ECD polypeptide and the fusion partner are noncovalently linked. In some such embodiments, they may be linked, for example, using binding pairs.
• Exemplary binding pairs include, but are not limited to, biotin and avidin or streptavidin, an antibody and its antigen, etc.
• signal peptide refers to a sequence of amino acid residues located at the N terminus of a polypeptide that facilitates secretion of a polypeptide from a mammalian cell.
• a signal peptide may be cleaved upon export of the polypeptide from the mammalian cell, forming a mature protein.
• Signal peptides may be natural or synthetic, and they may be heterologous or homologous to the protein to which they are attached.
• Exemplary signal peptides include, but are not limited to, FGFRl signal peptides, such as, for example, the amino acid sequence of SEQ ID NO: 7.
• Exemplary signal peptides also include signal peptides from heterologous proteins.
• polynucleotide is referred to as "isolated" when it is not part of the larger polynucleotide (such as, for example, genomic DNA or mitochondrial DNA, in the case of a DNA polynucleotide) in which it is typically found in nature, or is separated from at least some of the components of the cell in which it was produced, e.g., in the case of an RNA
• therapeutic agents include, but are not limited to, e.g., chemotherapeutic agents, growth inhibitory agents, cytotoxic agents, agents used in radiation therapy, anti-angiogenic agents, apoptotic agents, anti-tubulin agents, and other agents to treat cancer, such as anti-VEGF antibodies (e.g., bevacizumab, AVASTIN ® ), anti-HER-2 antibodies (e.g., trastuzumab, HERCEPTIN ® ), anti-CD20 antibodies (e.g., rituximab, RITUXAN ® ), an epidermal growth factor receptor (EGFR) antagonist (e.g., a tyrosine kinase inhibitor), HER 1 /EGFR inhibitors (e.g., erlotinib, TARCEVA ® ), platelet derived growth factor inhibitors (e.g., GLEEVEC ® , imatinib mesylate)), COX
• anti-VEGF antibodies
• etoglucid gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine;
• paclitaxel e.g., TAXOL®
• albumin-engineered nanoparticle formulation of paclitaxel e.g.,
• ABRAXANETM ABRAXANETM
• docetaxel e.g., TAXOTERE®
• chloranbucil 6-thioguanine
• mercaptopurine mercaptopurine
• methotrexate platinum agents such as cisplatin, oxaliplatin (e.g.,
• an anti-angiogenic agent is an antibody or other antagonist to an angiogenic agent as defined above, e.g., fusion proteins that binds to VEGF- A such as ZALTRAPTM (Aflibercept), antibodies to VEGF-A such as AVASTI ®
• VEGF antagonist specifically includes molecules, including antibodies, antibody fragments, other binding polypeptides, peptides, and non-peptide small molecules, that bind to VEGF and are capable of neutralizing, blocking, inhibiting, abrogating, reducing or interfering with VEGF activities.
• VEGF activities specifically includes VEGF mediated biological activities of VEGF.
• a cell with FGFRl gene amplification comprises at least 3 copies, at least 4 copies, at least 5 copies, at least 6 copies, at least 8 copies, or at least 10 copies of the FGFRl gene. In some embodiments, a cell with FGFRl gene amplification comprises at least 4 copies. In some embodiments, a cell with FGFRl gene amplification has a ratio of FGFRl gene: chromosome 8 centromere of at least 1.5, at least 2, at least 2.5, at least 3, at least 3.5, or at least 4. In some embodiments, a cell with FGFRl gene
• a cell with FGFR3 gene amplification comprises at least 3 copies, at least 4 copies, at least 5 copies, at least 6 copies, at least 8 copies, or at least 10 copies of the FGFR3 gene. In some embodiments, a cell with FGFR3 gene amplification comprises at least 4 copies. In some embodiments, a cell with FGFR3 gene amplification has a ratio oiFGFR3 gene: chromosome 4 centromere of at least 1.5, at least 2, at least 2.5, at least 3, at least 3.5, or at least 4. In some embodiments, a cell with FGFR3 gene
• a cell with FGF2 gene amplification comprises at least 3 copies, at least 4 copies, at least 5 copies, at least 6 copies, at least 8 copies, or at least 10 copies of the FGF2 gene. In some embodiments, a cell with FGF2 gene amplification comprises at least 4 copies. In some embodiments, a cell with FGF2 gene amplification has a ratio oiFGF2 gene:chromosome 4 centromere of at least 1.5, at least 2, at least 2.5, at least 3, at least 3.5, or at least 4. In some embodiments, a cell with FGF2 gene amplification has a ratio oiFGF2 gene:chromosome 4 centromere of at least 2.
• a "cell with FGF2 overexpression” or a “cell that overexpresses FGF2” refers to a cell that has at least a 2-fold greater level of FGF2 mRNA or protein than a reference cell.
• a “cancer with FGF2 overexpression” or a “cancer that overexpresses FGF2” refers to a cancer in which at least a portion of the cells have at least a 2-fold greater level of FGF2 mRNA or protein than a reference cell.
• a cell with FGF2 refers to a cancer in which at least a portion of the cells have at least a 2-fold greater level of FGF2 mRNA or protein than a reference cell.
• overexpression has at least 3-fold, at least 4-fold, at least 5-fold, at least 7-fold, or at least 10- fold greater level of FGF2 mRNA or protein than a reference cell.
• the level of FGF2 mRNA or protein can be determined by any suitable method including, but not limited to, the methods described herein.
• An exemplary human FGF2 protein sequence can be found, e.g., at NCBI Reference Sequence: NP_001997.5 dated 12-FEB-2012.
• An exemplary human FGF2 mRNA sequence can be found, e.g., at NCBI Reference Sequence: NM_002006.4 dated 12-FEB-2012.
• a "cell with DKK3 overexpression” or a “cell that overexpresses DKK3” refers to a cell that has at least a 2-fold greater level of DKK3 mRNA or protein than a reference cell.
• a “cancer with DKK3 overexpression” or a “cancer that overexpresses DKK3” refers to a cancer in which at least a portion of the cells have at least a 2-fold greater level of DKK3 mRNA or protein than a reference cell.
• a cell with DKK3 overexpression has at least 3-fold, at least 4-fold, at least 5-fold, at least 7-fold, or at least 10- fold greater level of DKK3 mRNA or protein than a reference cell.
• a cancer is considered to be ER positive when >1% of the tumor cell nuclei are immunoreactive in an immunohistochemistry (IHC) assay for the estrogen receptor.
• IHC immunohistochemistry
• a cancer is considered to be ER positive according to an assay manufacturer's or assay laboratory's guidelines.
• a "cancer that is HER2 positive” refers to a cancer that has been determined to be HER2 positive.
• a cancer that has been determined to be HER2 positive using an immunohistochemistry (IHC) assay for the HER2 protein, and/or a fluorescent in situ hybridization (FISH) assay to detect HER2 gene amplification.
• IHC immunohistochemistry
• FISH fluorescent in situ hybridization
• a cancer is characterized as HER2 positive when the IHC cell membrane stain intensity is 3+ on a scale from 0 to 3+.
• a HER2 FISH assay is used to determine whether the HER2 gene is amplified.
• the HER2 gene is considered to be amplified when the ratio of copies of the HER2 gene to chromosome 17 centromere is greater than 2. In some embodiments, if the HER2 gene is amplified, the breast cancer is considered to be HER2 positive, regardless of the results of an IHC assay. In some embodiments, a cancer is considered to be HER2 positive according to an assay
• a cancer with HER2 gene amplification refers to a cancer in which at least a portion of the cancer cells have an HER2 gene:chromosome 17 centromere ratio of greater than 1.
• An exemplary HER2 gene sequence can be found, e.g., NCBI Reference Sequence: NG_007503.1 dated 22-APR-2013.
• HER2 gene amplification is determined according to Persons, et al. "Fluorescence in situ hybridization (FISH) for detection of HER-2/neu amplification in breast cancer: a multicenter portability study.” Ann Clin Lab Sci 30: 41-48 (2000), which is incorporated by reference herein in its entirety for any purpose.
• a cell with HER2 gene amplification has a ratio oiHER2 gene:chromosome 17 centromere of greater than 2.
• each copy of the HER2 gene in a cell with HER2 gene amplification need not be a complete copy of the HER2 gene.
• a cell with HER2 gene amplification has elevated levels of HER2 (i.e., in some embodiments, a cell with HER2 gene amplification is also a cell with HER2 overexpression).
• a cancer in which at least a portion of the cells have HER2 gene amplification and/or HER2 overexpression is considered to be HER2 positive.
• a cancer is determined to be p95HER2 positive by IHC. In some such embodiments, a cancer is determined to be p95HER2 positive using the methods described insperinde et al, Clin. Cane. Res., 2010, 16(16): 4226-4235, such as methods using anti-p95 antibody clone D9 in a VeraTag assay. In some embodiments, a cancer is determined to be p95HER2 positive using the methods described in U.S. Patent No. 8,389,227 B2, such as methods using an antibody produced by a hybridoma cell line deposited with the Deutschland Sammlung von Mikroorganismen und Zellen under accession number DSM ACC2904 or DSM ACC2980.
• an "aromatase inhibitor” refers to a molecule capable of neutralizing, blocking, inhibiting, abrogating, reducing or interfering with aromatase activities including, but not limited to, its ability to convert androgens (such as testosterone and androstenedione) into estrogens (such as estradiol and estrone).
• Gonadotropin-releasing hormone agonist and “GnRH agonist” refer to a molecule capable of stimulating or enhancing gonadotropin-releasing hormone receptor activities, including, but not limited to, eliciting release of luteinizing hormone (LH) and/or follicle-stimulating hormone (FSH) from the pituitary.
• LH luteinizing hormone
• FSH follicle-stimulating hormone
• Gonadotropin-releasing hormone antagonist and “GnRH antagonist” refer to a molecule capable of neutralizing, blocking, inhibiting, abrogating, reducing or interfering with gonadotropin-releasing hormone activities including, but not limited to, eliciting release of luteinizing hormone (LH) and/or follicle-stimulating hormone (FSH).
• Nonlimiting exemplary gonadotropin-releasing hormone antagonists include cetrorelix (e.g., Cetrotide®), ganirelix (e.g., Antagon®), abarelix (e.g., Plenaxis®), and degarelix (e.g., Firmagon®).
• 17-hydroxylase inhibitor refers to a molecule capable of neutralizing, blocking, inhibiting, abrogating, reducing or interfering with cytochrome P450 17A1 (alro referred to as steroid 17-alpha-monooxygenase) activities including, but not limited to, its ability to add a hydroxyl group to carbon 17 of the steroid D ring of pregnenolone or progesterone.
• a nonlimiting exemplary 17-hydroxylase inhibitor is abiraterone acetate (e.g., Zytiga®).
• Desirable effects of treatment include preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.
• a therapeutically effective amount is also one in which any toxic or detrimental effects of the FGFR1 fusion proteins are outweighed by the therapeutically beneficial effects.
• the effective amount of the drug may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and typically stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow to some extent and typically stop) tumor metastasis; inhibit, to some extent, tumor growth; allow for treatment of the tumor, and/or relieve to some extent one or more of the symptoms associated with the disorder.
• the drug may prevent growth and/or kill existing cancer cells, it may be cytostatic and/or cytotoxic.
• the terms “benefit”, “clinical benefit”, “responsiveness”, and “therapeutic responsiveness” as used herein in the context of benefiting from or responding to administration of a therapeutic agent, can be measured by assessing various endpoints, e.g., inhibition, to some extent, of disease progression, including slowing down and complete arrest; reduction in the number of disease episodes and/or symptoms; reduction in lesion size; inhibition (i.e., reduction, slowing down or complete stopping) of disease cell infiltration into adjacent peripheral organs and/or tissues; inhibition (i.e.
• a "pharmaceutically acceptable carrier” refers to a non-toxic solid, semisolid, or liquid filler, diluent, encapsulating material, formulation auxiliary, or carrier conventional in the art for use with a therapeutic agent that together comprise a "pharmaceutical composition" for administration to a subject.
• a pharmaceutically acceptable carrier is nontoxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation.
• the pharmaceutically acceptable carrier is appropriate for the formulation employed.
• the carrier may be a gel capsule. If the therapeutic agent is to be administered subcutaneously, the carrier ideally is not irritable to the skin and does not cause injection site reaction.
• a method of treating cancer having FGFRl gene amplification, FGFRl overexpression, FGFR3 gene amplification, FGFR3 overexpression, FGF2 overexpression, and/or FGF2 gene amplification comprises administering a therapeutically effective amount of an FGFRl ECD or an FGFRl ECD fusion molecule to the subject.
• Vantas® goserelin acetate (e.g., Zoladex®), deslorelin (e.g., Suprelorin®, Ovuplant®), nafarelin (e.g., Synarel®), and triptorelin.
• goserelin acetate e.g., Zoladex®
• deslorelin e.g., Suprelorin®, Ovuplant®
• nafarelin e.g., Synarel®
• triptorelin triptorelin.
• the subject with prostate cancer has previously been administered, or is currently being administered, diethylstilbestrol (DES). That is, in some embodiments, the subject with prostate cancer previously underwent therapy with DES, but completed or terminated that therapy prior to being administered a therapeutically effective amount of an FGFRl ECD or an FGFRl ECD fusion molecule. In some embodiments, the subject with prostate cancer receives DES therapy concurrently with FGFRl ECD or FGFRl ECD fusion molecule therapy. By “concurrently” it is meant that there is a time period during which both (or all) agents exert their biological activities.
• DES diethylstilbestrol
• methods of treating ovarian cancer in a subject comprising administering a therapeutically effective amount of an FGFRl ECD or an FGFRl ECD fusion molecule to a subject with ovarian cancer.
• the ovarian cancer has been determined to have FGFRl gene amplification, FGFRl
• the FGFR1 ECD has an amino acid sequence selected from SEQ ID NOs: 1 to 4. In some embodiments, the FGFR1 ECD has an amino acid sequence selected from SEQ ID NOs: 2 and 4. In some embodiments, the FGFR1 ECD fusion molecule has an amino acid sequence selected from SEQ ID NOs: 5 and 6. In some embodiments, the FGFRl ECD fusion molecule is FGFRl ECD.339-Fc with an amino acid sequence of SEQ ID NO: 6.
• the subject's cancer has previously been treated, or is currently being treated, with one or more chemotherapeutic agents.
• chemotherapeutic agents may be used in the combined treatment methods and uses of the invention.
• An exemplary and non- limiting list of chemotherapeutic agents contemplated is provided herein under "Definitions" and in the "Summary of the Invention.”
• the invention provides methods of treating cancer, by administering therapeutically effective amounts of an FGFRl ECD and/or FGFRl ECD fusion molecule and one or more anti-angiogenic agent(s) to a subject.
• methods of treating cancer comprising administering to a subject an FGFRl ECD and/or FGFRl ECD fusion molecule in combination with at least one additional therapeutic agent selected from docetaxel, paclitaxel, vincristine, carboplatin, cisplatin, oxaliplatin, doxorubicin, 5-fluorouracil (5-FU), leucovorin, pemetrexed, sorafenib, etoposide, topotecan, a VEGF antagonist, an anti-VEGF antibody, a VEGF trap, and bevacizumab are provided.
• additional therapeutic agent selected from docetaxel, paclitaxel, vincristine, carboplatin, cisplatin, oxaliplatin, doxorubicin, 5-fluorouracil (5-FU), leucovorin, pemetrexed, sorafenib, etoposide, topotecan, a VEGF antagonist, an anti
• the FGFRl ECD and/or FGFRl ECD fusion molecule (e.g., FGFR1-ECD.339-Fc) is administered at a dose of about 10 mg/kg body weight, about 1 1 mg/kg body weight, about 12 mg/kg body weight, about 13 mg/kg body weight, about 14 mg/kg body weight, about 15 mg/kg body weight, about 16 mg/kg body weight, about 17 mg/kg body weight, about 18 mg/kg body weight, about 19 mg/kg body weight, or about 20 mg/kg body weight.
• the FGFRl fusion protein is administered at a dose of about 10 mg/kg body weight as calculated using an extinction coefficient of 1.1 1 mL/mg*cm.
• a dosage of 5 mg/kg using an EC of 1.1 1 mL/mg*cm herein corresponds to a dosage of about 3.9 mg/kg when calculated using an EC of 1.42 mL/mg*cm.
• a dosage of 10 mg/kg using an EC of 1.1 1 mL/mg*cm herein corresponds to a dosage of about 7.8 mg/kg when calculated using an EC of 1.42 mL/mg*cm.
• a dosage of 20 mg/kg using an EC of 1.11 mL/mg*cm herein corresponds to a dosage of about 15.6 mg/kg when calculated using an EC of 1.42 mL/mg*cm.
• compositions may be formulated for inhalation, for example, using pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen, and the like.
• the compositions may also be formulated, in various embodiments, into sustained release microcapsules, such as with biodegradable or nonbiodegradable polymers.
• a non-limiting exemplary biodegradable formulation includes poly lactic acid-glycolic acid polymer.
• a non-limiting exemplary non-biodegradable formulation includes a polyglycerin fatty acid ester. Certain methods of making such formulations are described, for example, in EP 1 125 584 Al .
• the instructions indicate that a ratio oiFGFR3 gene to chromosome 8 centromere of at least 2 in at least a portion of the cancer cells is indicative of therapeutic responsiveness to an FGFRl ECD and/or an FGFRl ECD fusion molecule. In some embodiments, the instructions indicate that a ratio oiFGFR3 gene to chromosome 8 centromere of greater than 2 in at least a portion of the cancer cells is indicative of therapeutic responsiveness to an FGFRl ECD and/or an FGFRl ECD fusion molecule.
• instructions includes, but is not limited to, labels, package inserts, instructions available in electronic form such as on a computer readable medium (e.g., a diskette, compact disk, or DVD), instructions available remotely such as over the internet, etc.
• a dosage pack is considered to include the instructions when the dosage pack provides access to the instructions, a link to the instructions (such as a uniform resource locator, or url), or other mechanism for obtaining a copy of the instructions (such as a return reply card, a physical address from which instructions may be requested, an e-mail address from which instructions may be requested, a phone number that may be called to obtain instructions, etc.).
• Nonlimiting exemplary FGFRl ECDs include full-length FGFRl ECDs, FGFRl ECD fragments, and FGFRl ECD variants.
• FGFRl ECDs may include or lack a signal peptide.
• Exemplary FGFRl ECDs include, but are not limited to, FGFRl ECDs having amino acid sequences selected from SEQ ID NOs.: 1, 2, 3, and 4.
• an FGFRl ECD comprises a sequence selected from SEQ ID NOs: 1 to 4. In some embodiments, an FGFRl ECD consists of a sequence selected from SEQ ID NOs: 1 to 4. When an FGFRl ECD "consists of a sequence selected from SEQ ID NOs: 1 to 4, the FGFRl ECD may or may not contain various post-translational
• an FGFRl ECD fusion molecule comprises a signal peptide. In some embodiments, an FGFRl ECD fusion molecule lacks a signal peptide. In some embodiments, the FGFRl ECD portion of an FGFRl ECD fusion molecule comprises a sequence selected from SEQ ID NOs: 1 to 4. In some embodiments, the FGFRl ECD portion of an FGFRl ECD fusion molecule consists of a sequence selected from SEQ ID NOs: 1 to 4.
• FGFRl ECD is linked to a fusion molecule
• the fusion partner portion of an FGFRl ECD fusion molecule is selected from Fc, albumin, and polyethylene glycol.
• Nonlimiting exemplary fusion partners are discussed herein.
• An FGFRl ECD fusion molecule may be prepared by attaching polyaminoacids or branch point amino acids to the FGFRl ECD.
• the polyaminoacid may be a carrier protein that serves to increase the circulation half life of the FGFRl ECD (in addition to the advantages achieved via a fusion molecule).
• such polyaminoacids should ideally be those that have or do not create neutralizing antigenic responses, or other adverse responses.
• Polymers for example, water soluble polymers, may be useful as fusion partners to reduce precipitation of the FGFR1 ECD fusion molecule in an aqueous environment, such as typically found in a physiological environment.
• Polymers employed in the invention will be pharmaceutically acceptable for the preparation of a therapeutic product or composition.
• polyethylene glycol is meant to encompass any of the forms that have been used to derivatize other proteins, such as mono-(Ci-Cio) alkoxy- or aryloxy- polyethylene glycol.
• Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water.
• Polymers used herein may be of any molecular weight and may be branched or unbranched.
• the polymers have an average molecular weight of between about 2 kDa to about 100 kDa (the term "about” indicating that in preparations of a polymer, some molecules will weigh more, some less, than the stated molecular weight).
• the average molecular weight of each polymer may be between about 5 kDa and about 50 kDa, or between about 12 kDa and about 25 kDa.
• the higher the molecular weight or the more branches the higher the higher the molecular weight or the more branches, the higher the
• Polymers of the invention are typically attached to a heterologous polypeptide at the alpha (a) or epsilon ( ⁇ ) amino groups of amino acids or a reactive thiol group, but it is also contemplated that a polymer group could be attached to any reactive group of the protein that is sufficiently reactive to become attached to a polymer group under suitable reaction conditions.
• a polymer may be covalently bound to an FGFR1 ECD via a reactive group, such as a free amino or carboxyl group.
• the amino acid residues having a free amino group may include lysine residues and the N-terminal amino acid residue.
• Those having a free carboxyl group may include aspartic acid residues, glutamic acid residues, and the C- terminal amino acid residue.
• Those having a reactive thiol group include cysteine residues.
• Methods for preparing fusion molecules conjugated with polymers will each generally involve (a) reacting an FGFR1 ECD with a polymer under conditions whereby the polypeptide becomes attached to one or more polymers and (b) obtaining the reaction product.
• Reaction conditions for each conjugation may be selected from any of those known in the art or those subsequently developed, but should be selected to avoid or limit exposure to reaction conditions such as temperatures, solvents, and pH levels that would inactivate the protein to be modified.
• the optimal reaction conditions for the reactions will be determined case-by -case based on known parameters and the desired result. For example, the larger the ratio of polymenpolypeptide conjugate, the greater the percentage of conjugated product.
• the optimum ratio in terms of efficiency of reaction in that there is no excess unreacted polypeptide or polymer
• the ratio of polymer (for example, PEG) to a polypeptide will generally range from 1 : 1 to 100 : 1.
• One or more purified conjugates may be prepared from each mixture by standard purification techniques, including among others, dialysis, salting-out, ultrafiltration, ion-exchange chromatography, gel filtration chromatography, and electrophoresis.
• FGFR1 ECDs of the present invention may be fused to marker sequences, such as a peptide that facilitates purification of the fused polypeptide.
• the marker amino acid sequence may be a hexa-histidine peptide such as the tag provided in a pQE vector (Qiagen, Mississauga, Ontario, Canada), among others, many of which are commercially available.
• pQE vector Qiagen, Mississauga, Ontario, Canada
• HA hemagglutinin
• oligomerization offers some functional advantages to a fusion protein, including, but not limited to, multivalency, increased binding strength, and the combined function of different domains.
• a fusion partner comprises an oligomerization domain, for example, a dimerization domain.
• the linker is selected based on the polynucleotide sequence that encodes it, to facilitate cloning the fusion partner and/or FGFRl ECD into a single expression construct (for example, a polynucleotide containing a particular restriction site may be placed between the polynucleotide encoding the fusion partner and the polynucleotide encoding the FGFRl ECD, wherein the polynucleotide containing the restriction site encodes a short amino acid linker sequence).
• linkers of various sizes may typically be included during the coupling reaction.
• Exemplary methods of non-covalently attaching a fusion partner to an FGFRl ECD include, but are not limited to, attachment through a binding pair.
• Exemplary binding pairs include, but are not limited to, biotin and avidin or streptavidin, an antibody and its antigen, etc.
• FGFR1 ECDs or FGFR1 ECD fusion molecules may be expressed in prokaryotic cells, such as bacterial cells; or in eukaryotic cells, such as fungal cells, plant cells, insect cells, and mammalian cells. Such expression may be carried out, for example, according to procedures known in the art.
• exemplary eukaryotic cells that may be used to express polypeptides include, but are not limited to, COS cells, including COS 7 cells; 293 cells, including 293 -6E cells; CHO cells, including CHO-S and DG44 cells; and NSO cells.
• FGFRl ECDs or FGFRl ECD fusion molecules may be purified by various methods known in the art. Such methods include, but are not limited to, the use of affinity matrices or hydrophobic interaction chromatography. Suitable affinity ligands include any ligands of the FGFRl ECD or of the fusion partner. Suitable affinity ligands in the case of an antibody that binds FGFRl include, but are not limited to, FGFRl itself and fragments thereof. Further, a Protein A, Protein G, Protein A/G, or an antibody affinity column may be used to bind to an Fc fusion partner to purify an FGFRl ECD fusion molecule.
• methods of identifying patients with cancer who may benefit from administration of an FGFRl ECD or FGFRl ECD fusion molecule are provided.
• the method comprises determining whether at least a portion of the cancer cells comprise FGFRl gene amplification, FGFRl overexpression, FGFR3 gene amplification, FGFR3 overexpression, FGF2 overexpression, and/or FGF2 gene
• FISH allows the determination of multiple parameters of gene amplification, including, but not limited to, the fraction of cells with an amplified gene, the amplification levels within various subpopulations of cells, and the amplification pattern within a cell (for example, a clustered signal versus multiple scattered signals).
• the ratio of the copy number of the gene of interest to the centromere reference for each cancer cell is determined.
• the mean ratio for a particular sample or subset of cells in a sample is then calculated. A mean ratio of greater than two is generally considered to indicate gene amplification, whereas signals between 1.5 to 2 may indicate low-level amplification.
• the tissue sample may be fixed (i.e. preserved) by conventional methodology (See e.g., "Manual of Histological Staining Method of the Armed Forces Institute of Pathology," 3 rd edition (1960) Lee G. Luna, HT (ASCP) Editor, The Blakston Division McGraw-Hill Book Company, New York; The Armed Forces Institute of Pathology Advanced Laboratory Methods in Histology and Pathology (1994) Ulreka V. Mikel, Editor, Armed Forces Institute of Pathology, American Registry of Pathology, Washington, D.C.).
• a fixative is determined by the purpose for which the sample is to be histologically stained or otherwise analyzed.
• the length of fixation depends upon the size of the tissue sample and the fixative used.
• neutral buffered formalin, Bouin's or paraformaldehyde may be used to fix a sample.
• the tissue sections are generally deparaffinized and rehydrated to water.
• the tissue sections may be deparaffinized by several conventional standard methodologies. For example, xylenes and a gradually descending series of alcohols may be used (See e.g., "Manual of Histological Staining Method of the Armed Forces Institute of Pathology", supra).
• deparaffinizing non-organic agents such as Hemo-De7 (CMS, Houston, Tex.) may be used.
• Fluorescent labels including, but are not limited to, rare earth chelates (europium chelates), Texas Red, rhodamine, fluorescein, dansyl, Lissamine, umbelliferone, phycocrytherin, phycocyanin, or commercially available fluorophores such SPECTRUM ORANGE7 and SPECTRUM GREEN7 and/or derivatives of any one or more of the above.
• the fluorescent labels can be conjugated to the antibody using the techniques disclosed in Current Protocols in Immunology, supra, for example, fluorescence can be quantified using a fluorimeter.
• Examples of enzyme-substrate combinations include, for example: (i) Horseradish peroxidase (HRPO) with hydrogen peroxidase as a substrate, wherein the hydrogen peroxidase oxidizes a dye precursor (e.g., orthophenylene diamine (OPD) or 3,3',5,5'- tetramethyl benzidine hydrochloride (TMB)); (ii) alkaline phosphatase (AP) with para- Nitrophenyl phosphate as chromogenic substrate; and (iii) .beta.-D-galactosidase (.beta.-D- Gal) with a chromogenic substrate (e.g., p-nitrophenyl-.beta.-D-galactosidase) or fluorogenic substrate (e.g., 4-methylumbelliferyl-.beta.-D-galactosidase).
• HRPO Horseradish peroxidase
• OPD ortho
• the label is indirectly conjugated with the antibody.
• the antibody can be conjugated with biotin and any of the four broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the antibody in this indirect manner.
• the antibody is conjugated with a small hapten and one of the different types of labels mentioned above is conjugated with an anti-hapten antibody.
• indirect conjugation of the label with the antibody can be achieved.
• the tissue section is exposed to primary antibody for a sufficient period of time and under suitable conditions such that the primary antibody binds to the target protein antigen in the tissue sample.
• Appropriate conditions for achieving this can be determined by routine experimentation.
• the extent of binding of antibody to the sample is determined by using any one of the detectable labels discussed above.
• the label is an enzymatic label (e.g. HRPO) which catalyzes a chemical alteration of the chromogenic substrate such as 3,3'-diaminobenzidine chromogen.
• the enzymatic label is conjugated to antibody which binds specifically to the primary antibody (e.g. the primary antibody is rabbit polyclonal antibody and secondary antibody is goat anti-rabbit antibody).
• Specimens thus prepared may be mounted and coverslipped. Slide evaluation is then determined, e.g., using a microscope, and staining intensity criteria, routinely used in the art, may be employed.
• a tiered system of staining is used to determine whether a cell or collection of cells overexpresses a protein.
• a four-tiered system is used in which the tiers are no staining (0), 1+, 2+, and 3+, where 1+, 2+, and 3+ indicate increasing levels of staining, respectively.
• greater than 1+, greater than 2+, or greater than 3+ may be used to indicate protein overexpression.
• a breast cancer is characterized as HER2 positive or HER2 negative according to IHC.
• a breast cancer is characterized as HER2 negative when the IHC cell membrane stain intensity is 0 or 1+.
• a breast cancer is characterized as HER2 positive when the IHC cell membrane stain intensity is 3+.
• the HER2 status of a breast cancer is equivocal when the IHC cell membrane stain intensity is 2+.
• a HER2 FISH assay is used to determine whether the HER2 gene is amplified. In some such embodiments, if the HER2 gene is amplified, the breast cancer is considered to be HER2 positive.
• the status of the estrogen receptor (ER) and/or progesterone receptor (PR) is determined according to the American Society of Clinical Oncology / College of American Pathologists Guideline Recommendations for Immunohistochemical Testing of Estrogen and Progesterone Receptors in Breast Cancer, J. Clin. Oncol, 2010, 28: 2784-2795 ("Guidelines”), which is incorporated by reference herein in its entirety for any purpose.
• the recommendations indicate that a breast cancer should be considered ER positive or PR positive when >1% of the tumor cell nuclei are immunoreactive in the corresponding IHC assay, and should be considered ER negative or PR negative when ⁇ 1% of tumor cell nuclei are immunoreactive in the corresponding IHC assay.
• Tissue or cell samples from mammals can be conveniently assayed for mRNAs using Northern, dot blot or PCR analysis.
• RT-PCR assays such as quantitative PCR assays are well known in the art.
• mRNA expression levels are levels quantified using real-time qRT-PCR.
• a method for detecting a target mRNA in a biological sample comprises producing cDNA from the sample by reverse transcription using at least one primer; amplifying the cDNA so produced using a target polynucleotide as sense and antisense primers to amplify target cDNAs therein; and detecting the presence of the amplified target cDNA.
• Optional methods of the invention include protocols which examine or detect mRNAs, such as target mRNAs, in a tissue or cell sample by microarray technologies.
• mRNAs such as target mRNAs
• test and control mRNA samples from test and control tissue samples are reverse transcribed and labeled to generate cDNA probes.
• the probes are then hybridized to an array of nucleic acids immobilized on a solid support.
• the array is configured such that the sequence and position of each member of the array is known.
• DNA microarrays are miniature arrays containing gene fragments that are either synthesized directly onto or spotted onto glass or other substrates. Thousands of genes are usually represented in a single array.
• a typical microarray experiment involves the following steps: 1) preparation of fluorescently labeled target from RNA isolated from the sample, 2) hybridization of the labeled target to the microarray, 3) washing, staining, and scanning of the array, 4) analysis of the scanned image and 5) generation of gene expression profiles.
• the perfect match probe has a sequence exactly complimentary to the particular gene and thus measures the expression of the gene.
• the mismatch probe differs from the perfect match probe by a single base substitution at the center base position, disturbing the binding of the target gene transcript. This helps to determine the background and nonspecific hybridization that contributes to the signal measured for the perfect match oligo.
• the Microarray Suite software subtracts the hybridization intensities of the mismatch probes from those of the perfect match probes to determine the absolute or specific intensity value for each probe set. Probes are chosen based on current information from Genbank and other nucleotide repositories. The sequences are believed to recognize unique regions of the 3' end of the gene.
• Microarray Suite software using preprogrammed fluidics protocols.
• the scanner is a confocal laser fluorescence scanner which measures fluorescence intensity emitted by the labeled cRNA bound to the probe arrays.
• the computer workstation with Microarray Suite software controls the fluidics station and the scanner.
• Microarray Suite software can control up to eight fluidics stations using preprogrammed hybridization, wash, and stain protocols for the probe array.
• the software also acquires and converts hybridization intensity data into a presence/absence call for each gene using appropriate algorithms.
• the software detects changes in gene expression between experiments by comparison analysis and formats the output into .txt files, which can be used with other software programs for further data analysis.
• Percentage tumor growth inhibition by FGFR1-ECD.339-Fc was determined by area-under-the-curve (AUC) analysis of xenograft growth curves treated with FGFR1- ECD.339-Fc compared to albumin control.
• FIG. 1 shows a scatterplot of the results of this analysis. Lung cancer xenografts with FGFR1 gene amplification had an average a 56% reduction in tumor growth with FGFR1-ECD.339-Fc treatment.
• lung cancer xenografts without FGFR1 gene amplification displayed an average 22% decrease in xenograft growth with FGFR1-ECD.339-Fc treatment compared to control.
• FIG. 4 shows a scatterplot of FGFRl mRNA expression comparing FGFRl gene amplified to non-amplified lung cancer xenografts.
• P 0.0146
• a sub- population of lung cancer xenograft models has high FGFRl RNA expression in the absence of FGFRl gene amplification.
• Xenograft models NCI-H226, NCI-H522 and PDX D35087 represent the 3 outlier points for FGFRl RNA expression in the non-amplified lung models (FIG. 4), with Gt/Sii-normalized gene expression levels of 3.70, 3.75 and 4.30, respectively.
• Tumor sizes were measured in each mouse on days 26, 35, 41 and 45 following the day of PDX D35087 implantation. The length and width of each tumor was measured using calipers and the tumor size calculated according to the formula:
• Tumor size (mm 3 ) (width (mm) x length (mm)) 2 /2
• RNA expression of a panel of FGFR1 -related genes including FGF ligands, FGF receptors, FGF binding proteins, FGF signaling molecules, and a group of angiogenesis- related targets was determined in a set of 35 tumor cell lines and xenografts using qRT-PCR.
• RNA was extracted from cell lines grown in vitro or tumor xenografts grown in vivo using the RNAeasy® mini kit (Qiagen, Germany). Extracted RNA was treated with DNAse I prior to creating cDNA with random hexamer priming and reverse transcriptase using the
• FIG. 8 shows anti-tumor activity of FGFR1-ECD.339-Fc in selected xenograft models. Representative tumor growth curves are shown for a renal cancer, Caki-1, (A), and mesothelioma, MSTO-211H, (B) xenograft cancer model.
• ⁇ -values are determined by a Mann- Whitney test of PCR gene expression in responders vs. non-responders for each gene using all models in Table 3.
• Table 5 Statistical analysis of FGF-related gene expression in relation to FGFR1- ECD.339-Fc anti-tumor response in non-FGFRl amplified lung xenograft models
• ⁇ -values are determined by a Mann- Whitney test of PCR gene expression in responders vs. non-responders for each gene using the non-FGFRl amplified lung models in table 5.
• FIG. 6 shows (A) FGF2 mRNA (normalized to GUSB) and (B) FGF2 protein expression in FGFR1-ECD.339-Fc responder and non-responder xenografts.
• FGF2 displayed a high ratio (247.7-fold) of mRNA gene expression between FGFR1-ECD.339-Fc responder and non-responder xenografts.
• FGF2 protein levels were also confirmed to correlate with FGFR1-ECD.339-Fc response.
• the tumor xenografts used in this experiment are shown in Table 7. Also shown in Table 7 are the dosing schedule for FGFR1-ECD.339-Fc in a mouse xenograft model, the percent tumor growth inhibition (TGI (%)) and the statistical significance of the tumor growth inhibition (P Value).
• Example 5 FGFR1-ECD.339-Fc mediated inhibition of FGF-2 and VEGF-A induced angiogenesis in a matrigel plug assay
• Recombinant human FGF-2 final concentration 250 ng/ml; Peprotech
• recombinant human VEGF-A final concentration 100 ng/ml; Peprotech
• matrigel BD Biosciences, Franklin Lakes, NJ
• sodium heparin 2 units/ml; Sigma
• FGF-2 and/or VEGF-A containing matrigel plugs were implanted subcutaneous ly in the abdomen region of C57BL/6 mice (Charles River, Wilmington, MA).
• FGFR1-ECD.339-Fc was administered by tail vein injection on days 1, 4, and 7 post-matrigel implantation. On day 9, plugs were excised and processed for hematoxylin and eosin (H&E) staining. Digital images of the stained matrigel sections were generated using a Retiga 2000R digital camera (Qlmaging, Burnaby, BC). Image analysis was performed using Image-Pro
• Neovascularization was defined as the cellular response in the Matrigel plugs, consisting of newly formed blood vessels and migrated cells.
• FIG. 10 The results of that experiment are shown in FIG. 10. Administration of 5 mg/kg or higher FGFR1-ECD.339-Fc completely blocked in vivo angiogenesis induced by a matrigel plug impregnated with FGF-2. Administration of 15 or 45 mg/kg FGFR1-ECD.339-Fc also completely blocked in vivo angiogenesis in response to a matrigel plug impregnated with VEGF-A only or FGF-2 plus VEGF-A.
• Anti-angiogenic activity against VEGF induced angiogenesis in this model system may reflect inhibition of the synergistic activity between VEGF in the plug and murine-derived stromal FGFs since SPR analysis shows that FGFR1- ECD.339-Fc does not directly interact with VEGF-A.
• HUVEC cells (Life Technologies, Grand Island, NY) were seeded at a density of 4X10 3 cells/well in basal media (Medium 200 (Life Technologies) with 2% heat inactivated FBS) and stimulated with either 10 ng/ml FGF2 (R&D Systems, Minneapolis,
• HUVEC cell proliferation was determined 3 days post-stimulation using CellTiter-Glo® Luminescent Cell Viability Assay.
• Example 6 FGFR1-ECD.339-Fc -mediated inhibition of FGFR1 signaling in the JIMT-1 breast cancer xenograft model
• FGFR1-ECD.339-Fc was detected using anti-human Fc monoclonal antibody (Jackson Immuno Research).
• a Phase 1 first-time-in-human study (Study FP 1039-001) has been completed.
• the study enrolled 39 subjects who received doses ranging from 0.6 mg/kg to 20.5 mg/kg
• Squamous NSCLC subjects who have documented tumor progression (based on radiological imaging) after receiving two or more prior lines of systemic therapy (including platinum containing chemotherapy regimens) for Stage IV disease may be enrolled. See, e.g., TNM Classification of Malignant Tumors, 7 th edition, Sobin et al, Eds., 2009; Edge et al., 2010, Ann. Surg. Oncol, 17: 1471-1474.
• Subjects with ER positive breast cancer having disease progression while on aromatase inhibitor therapy are allowed to continue aromatase inhibitor therapy, subjects with prostate cancer may continue to be treated with GnRH agonists or GnRH antagonists as clinically appropriate, and subjects with carcinoid cancer may continue treatment with octreotide.
• Exclusion criteria include treatment with any anti-cancer therapy (for biological anti-cancer therapies see additional exclusion criteria herein) during the preceding 4 weeks or within 4 half-lives of the therapy, whichever is longer (except: anti-cancer hormonal treatment of prostate cancer, breast cancer or octreotide for treatment of carcinoid cancer), receipt of any biological therapy within 6 weeks of the first dose of FGFR1-ECD.339-Fc, conditions likely to increase the potential for abdominal perforation or fistula formation, symptomatic leptomeningeal or brain metastases or spinal cord compression.
• EC 1.11 mL/mg*cm). In certain circumstances, subjects will receive FGFR1-ECD.339-Fc at the starting dose of 5 mg/kg, 10 mg/kg, or 15 mg/kg.
• At least 12 subjects with stage IV squamous non-small cell lung cancer (according to TNM Classification of Malignant Tumors, 7 th edition, Sobin et al, Eds., 2009; Edge et al., 2010, Ann. Surg. Oncol, 17: 1471-1474); and up to 30 subjects will be enrolled at the target dose to further evaluate safety and efficacy.
• the first cycle of chemotherapy may be initiated while subjects are still in screening for the present study.
• the first dose of FGFR1-ECD.339-Fc should be given no later than Cycle 2 Day 1 of
• a total of 4 to 6 cycles of paclitaxel/carboplatin will be administered per local clinical practice.
• the maximum dose is based on a GFR estimate that is capped at 125 mL/min for patients with normal renal function. No higher estimated GFR values should be used.
• the Cockroft-Gault formula (see below) can be used to calculate the creatinine clearance (CLCR), which can be substituted for the GFR in the Calvert formula.
• At least 12 subjects with stage IV squamous non-small cell lung cancer (according to TNM Classification of Malignant Tumors, 7 th edition, Sobin et al, Eds., 2009; Edge et al., 2010, Ann. Surg. Oncol, 17: 1471-1474); and up to 30 subject will be enrolled at the target dose to further evaluate safety and efficacy.
• Subjects will receive FGFR1-ECD.339-Fc administered as a 30-minute infusion once a week (Day 1, Day 8, and Day 15) of each 21 -day cycle at the dosages specified in Table 9. Following infusion of FGFR1-ECD.339-Fc, subjects should be observed for 1 hour prior to infusion of chemotherapeutic agents. If infusion reactions are noted, subjects should be treated with antiemetics, steroids, or antihistamines at the discretion of the investigator and premedication according to institutional standards before further infusions of FGFR1- ECD.339-Fc should be considered.
• Subjects in Arm B will receive pre-treatment for docetaxel according to institutional standards. Docetaxel will be administered according to the dose level being explored as described in Table 9 as an i.v. infusion over 1 hour (or according to local clinical standards) on Day 1 of each 21 day cycle. The subject is treated until progression or until the subject has been determined to have received maximum benefit.
• product labeling e.g. US package insert or product monograph.
• Table 10 lists certain sequences discussed herein. FGFRl sequences are shown without the signal peptide, unless otherwise indicated.
• MWSWKCLLFW AVLVTATLCT ARPSPTLPEQ AQPWGAPVEV ESFLVHPGDL LQLRCRLRDD VQSINWLRDG VQLAESNRTR ITGEEVEVQD SVPADSGLYA CVTSSPSGSD TTYFSVNVSD
• MWSWKCLLFW AVLVTATLCT ARPSPTLPEQ AQPWGAPVEV ESFLVHPGDL LQLRCRLRDD VQSINWLRDG VQLAESNRTR ITGEEVEVQD SVPADSGLYA CVTSSPSGSD TTYFSVNVSD ALPSSEDDDD DDDSSSEEKE TDNTKPNPVA PYWTSPEKME KKLHAVPAAK TVKFKCPSSG TPNPTLRWLK NGKEFKPDHR IGGYKVRYAT WSIIMDSWP SDKGNYTCIV ENEYGSINHT
• YQLDWERSP HRPILQAGLP ANKTVALGSN VEFMCKVYSD PQPHIQWLKH IEVNGSKIGP DNLPYVQILK TAGVNTTDKE MEVLHLRNVS FEDAGEYTCL AGNSIGLSHH SAWLTVLEAL EPKSSDKTHT CPPCPAPELL GGPSVFLFPP KPKDTLMISR TPEVTCVWD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRWSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT ISKAKGQPRE PQVYTLPPSR DELTKNQVSL TCLVKGFYPS DIAVEWESNG QPENNYKTTP PVLDSDGSFF LYSKLTVDKS RWQQGNVFSC SV HEALHNH YTQKSLSLSP GK
• EPKSSDKTHT CPPCPAPELL GGPSVFLFPP KPKDTLMISR TPEVTCVWD VSHEDPEVKF NWYVDGVEVH NAKTKPREEQ YNSTYRWSV LTVLHQDWLN GKEYKCKVSN KALPAPIEKT
• Exemplary Fc #2 AKGQPREPQV YTLPPSQEEM TKNQVSLTCL VKGFYPSDIA

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