Abstract:
The invention provides methods and compositions for use in treating diseases associated with excessive cellular proliferation, such as cancer.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application is a continuation-in-part of, and claims priority from, U.S. patent application Ser. No. 10/687,526, filed Oct. 16, 2003 (pending), which is a continuation of U.S. patent application Ser. No. 10/272,167, filed Oct. 16, 2002 (now U.S. Pat. No. 6,653,341), which is a continuation-in-part of, and claims priority from, U.S. patent application Ser. No. 09/843,617, filed Apr. 26, 2001 (now U.S. Pat. No. 6,469,182), which is a continuation of U.S. patent application Ser. No. 09/677,485, filed Oct. 2, 2000 (now U.S. Pat. No. 6,365,759), which is a continuation of U.S. patent application Ser. No. 09/334,488, filed Jun. 16, 1999 (now U.S. Pat. No. 6,214,865), which claims priority from U.S. Provisional Patent Application Ser. No. 60/089,682, filed Jun. 17, 1998 (now abandoned). The contents of the earlier filed applications are incorporated by reference herein in their entirety. 
    
    
     FIELD OF THE INVENTION 
     This invention relates to methods and compositions for use in treating cancer. 
     BACKGROUND OF THE INVENTION 
     Cancer is a term used to describe a wide variety of diseases that are each characterized by the uncontrolled growth of a particular type of cell. It begins in a tissue containing such a cell and, if the cancer has not spread to any additional tissues at the time of diagnosis, may be treated by, for example, surgery, radiation, or another type of localized therapy. However, when there is evidence that cancer has metastasized from its tissue of origin, different approaches to treatment are typically used. Indeed, because it is not possible to determine the extent of metastasis, systemic approaches to therapy are usually undertaken when any evidence of spread is detected. These approaches typically involve the administration of chemotherapeutic drugs that interfere with the growth of rapidly dividing cells, such as cancer cells. 
     Halichondrin B is a structurally complex, macrocyclic compound that was originally isolated from the marine sponge  Halichondria okadai , and subsequently was found in  Axinella  sp.,  Phakellia carteri , and  Lissondendryx  sp. A total synthesis of halichondrin B was published in 1992 (Aicher et al., J. Am. Chem. Soc. 114:3162-3164, 1992). Halichondrin B has been shown to inhibit tubulin polymerization, microtubule assembly, beta S -tubulin crosslinking, GTP and vinblastine binding to tubulin, and tubulin-dependent GTP hydrolysis in vitro. This molecule has also been shown to have anti-cancer properties in vitro and in vivo. Halichondrin B analogs having anti-cancer activities are described in U.S. Pat. No. 6,214,865 B1. 
     SUMMARY OF THE INVENTION 
     The invention provides methods of inhibiting growth of tumors in patients, which involve administering to the patients a compound of the structure: 
                                
or a pharmaceutically acceptable salt thereof, wherein the tumor is breast carcinoma, and wherein the compound or pharmaceutically acceptable salt is not co-administered with another pharmaceutically active agent. The compound or pharmaceutically acceptable salt may be administered in a pharmaceutical composition including a pharmaceutically-acceptable carrier. The compound or pharmaceutically acceptable salt is administered, for example, by oral, topical, parenteral, or intravenous routes, or by injection or inhalation. Alternatively, the compound or pharmaceutically acceptable salt is administered in a controlled-release formulation.
 
     The invention provides methods of treating cancer in patients, which involve administering to the patients a compound having the structure: 
                                
or a pharmaceutically acceptable salt thereof, in combination with one or more second anti-cancer agents selected from the group consisting of floxuridine, vinorelbine, and an antibody (e.g., a monoclonal antibody). In one example, the antibody is specific for HER-2/erb B2 (e.g., trastuzumab).
 
     The methods of the invention can be used in the treatment of, for example, non-small cell lung cancer, breast cancer, colorectal cancer, pancreatic cancer, ovarian cancer, skin cancer, prostate cancer, cancer of the brain or nervous system, head and neck cancer, testicular cancer, lung cancer, liver cancer, kidney cancer, bladder cancer, gastrointestinal cancer, bone cancer, cancer of the endocrine system, cancer of the lymphatic system, fibrosarcoma, neurectodermal tumor, mesothelioma, epidermoid carcinoma, or Kaposi&#39;s sarcoma. 
     The methods of the invention can also include administration of one or more additional anti-cancer agents. For example, agents such as antimetabolites (e.g., methotrexate, purine antagonists (e.g., mercaptopurine, thioguanine, fludarabine phosphate, cladribine, or pentostatin), or pyrimidine antagonists (e.g., gemcitabine, capecitabine, fluorouracil, 5-fluorouracil, cytarabine, or azacitidine)), antibiotics (e.g., anthracyclines (e.g., doxorubicin, epirubicin, daunorubicin, or idarubicin), adriamycin, dactinomycin, idarubincin, plicamycin, mitomycin, or bleomycin), alkylating agents (e.g., procarbazine, dacarbazine, altretamine, cisplatin, carboplatin, or nitrosoureas), plant alkaloids (e.g., vinblastine, vincristine, etoposide, teniposide, topotecan, irinotecan, paclitaxel, or docetaxel), anticoagulants (e.g., heparin or warfarin), antibodies, biological agents (e.g., hormonal agents, cytokines, interleukins, interferons, granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), chemokines, or vaccine antigens), and/or anti-angiogenic agents (e.g., angiostatin or endostatin) can be used. 
     The invention also provides compositions that include a compound having the structure: 
                                
or a pharmaceutically acceptable salt thereof, in combination with one or more second anti-cancer agents selected from the group consisting of floxuridine, vinorelbine, and an antibody (e.g., a monoclonal antibody). In one example, the antibody is specific for HER-2/erb B2 (e.g., trastuzumab).
 
     The compositions can include, optionally, one or more additional anti-cancer agents. For example, agents such as antimetabolites (e.g., methotrexate, purine antagonists (e.g., mercaptopurine, thioguanine, fludarabine phosphate, cladribine, or pentostatin), or pyrimidine antagonists (e.g., gemcitabine, capecitabine, fluorouracil, 5-fluorouracil, cytarabine, or azacitidine)), antibiotics (e.g., anthracyclines (e.g., doxorubicin, epirubicin, daunorubicin, or idarubicin), adriamycin, dactinomycin, idarubincin, plicamycin, mitomycin, or bleomycin), alkylating agents (e.g., procarbazine, dacarbazine, altretamine, cisplatin, carboplatin, or nitrosoureas), plant alkaloids (e.g., vinblastine, vincristine, etoposide, teniposide, topotecan, irinotecan, paclitaxel, or docetaxel), anticoagulants (e.g., heparin or warfarin), antibodies, biological agents (e.g., hormonal agents, cytokines, interleukins, interferons, granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), chemokines, or vaccine antigens), and/or anti-angiogenic agents (e.g., angiostatin or endostatin) can be included in the compositions. 
     The invention further provides kits that include a compound having the structure: 
                                
or a pharmaceutically acceptable salt thereof, and a second (or further) anti-cancer agent.
 
     Second anti-cancer agents of the kits of the invention can be, for example, antimetabolites (e.g., methotrexate, purine antagonists (e.g., mercaptopurine, thioguanine, fludarabine phosphate, cladribine, or pentostatin), or pyrimidine antagonists (e.g., gemcitabine, capecitabine, fluorouracil, 5-fluorouracil, floxuridine, cytarabine, or azacitidine)), antibiotics (e.g., anthracyclines (e.g., doxorubicin, epirubicin, daunorubicin, or idarubicin), adriamycin, dactinomycin, idarubincin, plicamycin, mitomycin, or bleomycin), alkylating agents (e.g., procarbazine, dacarbazine, altretamine, cisplatin, carboplatin, or nitrosoureas), plant alkaloids (e.g., vinblastine, vincristine, etoposide, teniposide, topotecan, irinotecan, vinorelbine, paclitaxel, or docetaxel), anticoagulants (e.g., heparin or warfarin), antibodies (e.g., monoclonal antibodies and/or antibodies specific for HER-2/erb B2 (e.g., trastuzumab)), biological agents (e.g., hormonal agents, cytokines, interleukins, interferons, granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), chemokines, or vaccine antigens), and/or anti-angiogenic agents (e.g., angiostatin or endostatin). 
     Other features and advantages of the invention will be apparent from the following detailed description and the claims. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The invention provides methods for treating cancer, involving administration of a halichondrin B analog, such as an analog having the following structure: 
                                
which is carried out in combination with a second approach to treatment. In addition to (or in place of) the compound noted above, any one or more of the halichondrin B analogs described in the patents noted above can be used in the invention.
 
     There are numerous types of anti-cancer approaches that can be used in conjunction with halichondrin B analog treatment, according to the invention. These include, for example, treatment with chemotherapeutic agents (see below), biological agents (e.g., hormonal agents, cytokines (e.g., interleukins, interferons, granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), and granulocyte macrophage colony stimulating factor (GM-CSF)), chemokines, vaccine antigens, and antibodies), anti-angiogenic agents (e.g., angiostatin and endostatin), radiation, and surgery. 
     The methods of the invention can employ these approaches to treat the same types of cancers as those for which they are known in the art to be used, as well as others, as can be determined by those of skill in this art. Also, these approaches can be carried out according to parameters (e.g., regimens and doses) that are similar to those that are known in the art for their use. However, as is understood in the art, it may be desirable to adjust some of these parameters, due to the additional use of a halichondrin B analog with these approaches. For example, if a drug is normally administered as a sole therapeutic agent, when combined with a halichondrin B analog, according to the invention, it may be desirable to decrease the dosage of the drug, as can be determined by those of skill in this art. Examples of the methods of the invention, as well as compositions and kits that can be used in these methods, are provided below. 
     Chemotherapeutic drugs of several different types including, for example, antimetabolites, antibiotics, alkylating agents, plant alkaloids, hormonal agents, anticoagulants, antithrombotics, and other natural products, among others, can be used in conjunction with halichondrin B treatment, according to the invention. Specific, non-limiting examples of these classes of drugs, as well as cancers that can be treated by their use, are as follows. 
     Antimetabolite drugs that halichondrin B analogs can be used with include, e.g., methotrexate, purine antagonists (e.g., mercaptopurine, thioguanine, fludarabine phosphate, cladribine, and pentostatin), and pyrimidine antagonists (e.g., gemcitabine, capecitabine, fluorouracil (e.g., 5-FU), floxuridine (FUdR; 2′-deoxy-5-fluoro-uridine; 5′-DFUR), cytarabine, and azacitidine). Use of these agents to treat particular types of cancers is well known in the art, and these agents can be used in combination with halichondrin B analogs to treat these and other types of cancers. As specific, non-limiting examples, a halichondrin B analog can be used with gemcitabine in the treatment of non-small cell lung carcinoma, pancreatic cancer, or metastatic breast cancer. In an additional example, a halichondrin B analog can be used in conjunction with capecitabine in the treatment of breast or colorectal cancers. In another example, floxuridine is used with a halichondrin B analog to treat breast cancer. 
     As is noted above, another class of chemotherapeutic drugs with which halichondrin B analogs can be used includes anticancer antibiotics. These include, for example, anthracyclines (e.g., doxorubicin, epirubicin, daunorubicin, and idarubicin), adriamycin, dactinomycin, idarubincin, plicamycin, mitomycin, and bleomycin. As with the drugs mentioned above, use of these agents to treat particular types of cancers is well known in the art, and they can be used in combination with halichondrin B analog treatment to treat these and other types of cancers. As a specific, non-limiting example, an anthracycline, such as doxorubicin, can be administered in conjunction with halichondrin B analog therapy for the treatment of breast or pancreatic cancers. Alternatively, a third agent, cyclophosphamide, can be used in this method. 
     Alkylating agents comprise another class of chemotherapeutic drugs that can be administered in conjunction with a halichondrin B analog, according to the invention. Examples of such drugs include procarbazine, dacarbazine, altretamine, cisplatin, carboplatin, and nitrosoureas. Halichondrin B analogs can be used with these agents in the treatment of cancers that these agents are known in the art to be used to treat, as well as in the treatment of other cancers. For example, a halichondrin B analog can be used in conjunction with carboplatinum in the treatment of non-small cell lung carcinoma or ovarian cancer. 
     An additional type of chemotherapeutic drug with which halichondrin B analogs can be administered, according to the invention, is plant alkaloids, such as vinblastine, vincristine, etoposide, teniposide, topotecan, irinotecan, vinorelbine, paclitaxel, and docetaxel. As specific, non-limiting examples, a halichondrin B analog can be used in conjunction with irinotecan for the treatment of colorectal cancer, or with topotecan in the treatment of ovarian or non-small cell lung cancers. In another example, a halichondrin B analog can be administered with vinorelbine in the treatment of breast cancer of non-small cell lung cancer. 
     Further types of anti-cancer agents that can be used in conjunction with halichondrin B analog treatment, according to the invention, are anticoagulants and antithrombotic agents. For example, heparin (e.g., low molecular weight heparin or heparin sulfate) or warfarin can be used. Use of these agents in treating patients by, for example, injection or oral administration, is well known in the art, and thus they can readily be adapted by those of skill in the art for use in the present invention. 
     Therapeutic antibodies can also be administered in conjunction with halichondrin B analog treatment, according to the invention. For example, antibodies (e.g., monoclonal antibodies) against tumor or cancer cell specific or enriched antigens can be used. As a specific example, antibodies against HER-2/erb B2, which is a protein that is overexpressed in breast cancer cells, can be used. Herceptin® (trastuzumab; Genentech, Inc.) is an example of a monoclonal antibody that is specific for HER-2/erb B2 that can be used in the invention. 
     Numerous approaches for administering anti-cancer drugs are known in the art, and can readily be adapted for use in the present invention. In the case of one or more drugs that are to be administered in conjunction with a halichondrin B analog, for example, the drugs can be administered together, in a single composition, or separately, as part of a comprehensive treatment regimen. For systemic administration, the drugs can be administered by, for example, intravenous infusion (continuous or bolus). Appropriate scheduling and dosing of such administration can readily be determined by those of skill in this art based on, for example, preclinical studies in animals and clinical studies (e.g., phase I studies) in humans. In addition, analysis of treatment using similar drugs, as well as monitoring factors such as blood counts (e.g., neutrophil and platelet counts) and vital signs in patients can be used, as is well understood in the art. 
     Many regimens used to administer chemotherapeutic drugs involve, for example, intravenous administration of a drug (or drugs) followed by repetition of this treatment after a period (e.g., 1-4 weeks) during which the patient recovers from any adverse side effects of the treatment. It may be desirable to use both drugs at each administration or, alternatively, to have some (or all) of the treatments include only one drug (or a subset of drugs). 
     As a specific, non-limiting example of a treatment regimen included in the invention, a halichondrin B analog (e.g., 0.01-5 mg/m 2 ) can be administered to a patient by intravenous infusion for 0.5-3 hours, followed by intravenous infusion of another drug (e.g., gemcitabine, e.g., 500-900 mg/m 2 ) for 0.5-3 hours. This course of treatment can be repeated every 2-3 weeks, as determined to be tolerable and effective by those of skill in the art. In a variation of this method, the treatment is carried out with both drugs on the first day, as is noted above, but then is followed up with treatment using only the secondary drug (e.g., gemcitabine) in ensuing weeks. 
     Further, as is well known in the art, treatment using the methods of the invention can be carried out in conjunction with the administration of antiemetics, which are drugs that are used to reduce the nausea and vomiting that are common side effects of cancer chemotherapy. Examples of such drugs include major tranquilizers (e.g., phenothiazines, such as chlorpromazine and prochlorperazine), dopamine antagonists (e.g., metoclopramide), serotonin antagonists (e.g., ondansetron and granisetron), cannabinoids (e.g., dronabinol), and benzodiazepine sedatives. 
     In addition to the cancers mentioned above, the methods and compositions of the invention can be used to treat the following types of cancers, as well as others: skin (e.g., squamous cell carcinoma, basal cell carcinoma, or melanoma), prostate, brain and nervous system, head and neck, testicular, lung, liver (e.g., hepatoma), kidney, bladder, gastrointestinal, bone, endocrine system (e.g., thyroid and pituitary tumors), and lymphatic system (e.g., Hodgkin&#39;s and non-Hodgkin&#39;s lymphomas) cancers. Other types of cancers that can be treated using the methods of the invention include fibrosarcoma, neurectodermal tumor, mesothelioma, epidermoid carcinoma, and Kaposi&#39;s sarcoma. 
     The invention also includes compositions that include a halichondrin B analog in combination with an additional therapeutic agent(s), such as any of those agents listed above. The drugs in these compositions preferably are formulated for administration to patients (e.g., in physiological saline) or, alternatively, can be in a form requiring further processing prior to administration. For example, the compositions can include the drugs in a lyophilized form or in a concentrated form requiring dilution. Formulation of drugs for use in chemotherapeutic methods can be carried out using standard methods in the art (see, e.g.,  Remington&#39;s Pharmaceutical Sciences  (18 th  edition), ed. A. Gennaro, 1990, Mack Publishing Co., Easton, Pa.). 
     Also included in the invention are kits that include one or more halichondrin B analogs and one or more additional anti-cancer agents, as is discussed above. The halichondrin B analog(s) and additional anti-cancer agent(s) can be present in a single container, such as a vial, or can be present in separate containers. Further, the different agents can be present in forms that are ready for administration or forms requiring further formulation (e.g., lyophilized form). The kits can also include diluents for the agents, instructions for administration of the agents, one or more labels listing the contents of the kits, and/or devices used in agent administration. 
     The disclosed compound has pharmacological activity, including anti-tumor and anti-mitotic activity, as demonstrated in section D of U.S. Pat. No. 6,214,865. Examples of tumors include melanoma, fibrosarcoma, monocytic leukemia, colon carcinoma, ovarian carcinoma, breast carcinoma, osteosarcoma, prostate carcinoma, lung carcinoma and ras-transformed fibroblasts. 
     The invention features pharmaceutical compositions that include a compound of formula (I) (see U.S. Pat. No. 6,214,865 for formula (I)) and a pharmaceutically-acceptable carrier. Compositions can also include a combination of disclosed compounds, or a combination of one or more disclosed compounds and other pharmaceutically-active agents, such as an anti-tumor agent, an immune-stimulating agent, an interferon, a cytokine, an anti-MDR agent or an anti-angiogenesis agent. Compositions can be formulated for oral, topical, parenteral, intravenous, or intramuscular administration, or administration by injection or inhalation. Formulations can also be prepared for controlled-release, including transdermal patches. 
     A method for inhibiting tumor growth in a patient includes the step of administering to the patient an effective, anti-tumor amount of a disclosed compound or composition. The invention also contemplates combination therapies, including methods of co-administering a compound of formula (I) (see U.S. Pat. No. 6,214,865 for formula (I)) before, during, or after administering another pharmaceutically active agent. The methods of administration may be the same or different. Inhibition of tumor growth includes a growth of the cell or tissue exposed to the test compound that is at least 20% less, and preferably 30%, 50%, or 75% less than the growth of the control (absence of known inhibitor or test compound).