JP2009509974A - Anti-cancer treatment - Google Patents

Abstract

Cancer can be treated by administration of glufosfamide alone or in combination with other anticancer agents. The present invention provides compositions and methods for treating cancer. In particular, the present invention relates to administration of glufosfamide alone or in combination with other anticancer agents for the treatment of cancer. Furthermore, the present invention provides methods for the treatment of advanced and / or metastatic malignancies, including those previously treated with chemotherapy. In a first aspect, the present invention provides a method for treating cancer, wherein the method is administered to a subject in need of such treatment, alone or in combination with gemcitabine or other anticancer agent, in a therapeutically effective amount of glufosfamide. Including doing. A wide variety of cancers such as breast cancer, colorectal cancer, ovarian cancer, lung cancer and pancreatic cancer can be treated by this method.

Description

(Related application)
This application claims the benefit of US Pat. No. 60 / 719,787 filed Sep. 23, 2005 and 60/760599, filed Jan. 20, 2006, the contents of which are hereby incorporated by reference. It is incorporated in the specification.

(Technical field)
The present invention provides compositions and methods for the treatment of cancer and thus relates to the fields of medicine and pharmacology.

(Background of the Invention)
“Cancer” is one of a group of more than 100 diseases caused by uncontrolled abnormal growth of cells that can generally expand towards nearby tissues or other parts of the body. Cancer cells can form solid tumors where the cancer cells agglomerate or exist as dispersed cells such as leukemia. Normal cells divide (regenerate) only as needed until maturation is achieved, and thereafter for replacement of damaged or dead cells. Cancer cells are often referred to as "malignant" because they divide indefinitely and eventually penetrate into nearby cells and spread to other parts of the body. Identify cancer cells from benign tumor cells that do not expand to other organs or parts of the growing body due to their tendency to expand from one organ to another or from one part of the body to another be able to. Malignant cancer cells eventually metastasize and expand to other parts of the body via the bloodstream or lymphatic system, where they grow and form new tumors. This type of tumor progression makes cancer a fatal disease. Although the diagnosis and treatment of tumors has advanced greatly, many humans die from cancer each year, and the death is typically due to cancer that is resistant to metastasis and conventional therapies. Current methods for the treatment of advanced and / or metastatic malignancies previously treated with chemotherapy (ie chemotherapy refractory cancer) are inadequate. Curative therapy is almost impossible in patients with advanced malignant disease that has recurred after chemotherapy.

  There is a need in the art for advanced methods for the treatment of advanced and / or metastatic malignancies previously treated with chemotherapy. The present invention focuses on such a need and provides compositions and methods for treating cancer.

(Summary of the Invention)
The present invention provides compositions and methods for treating cancer. In particular, the present invention relates to administration of glufosfamide alone or in combination with other anticancer agents for the treatment of cancer. Furthermore, the present invention provides methods for the treatment of advanced and / or metastatic malignancies, including those previously treated with chemotherapy.

  In a first aspect, the present invention provides a method for treating cancer, wherein the method is administered to a subject in need of such treatment, alone or in combination with gemcitabine or other anticancer agent, in a therapeutically effective amount of glufosfamide. Including doing. A wide variety of cancers such as breast cancer, colorectal cancer, ovarian cancer, lung cancer and pancreatic cancer can be treated by this method.

According to the methods of the present invention, glufosfamide is at least 1, 2, 3, 4, 5, 6, 7, regardless of whether administration of glufosfamide is performed alone or in combination with other anticancer agents such as gemcitabine. Administer at least once per treatment cycle for 8 or more treatment cycles, where each cycle is at least 1 week and can be as long as 3 or 4 weeks, or as long as 7 weeks, and each of glufosfamide In administration, glufosfamide is administered to cancer patients by intravenous infusion in amounts ranging from 1.5 to about 6.0 g / m ² (g is the grams of glufosfamide and m ² is the patient's surface area). . In one embodiment, the amount of glufosfamide administered ranges from about 1.5 to about 4.5 g / m ² . In one embodiment, the amount of glufosfamide administered is 4.5 g / m ² . In one embodiment, the amount of glufosfamide administered is 5.0 g / m ² . Infusion of glufosfamide can be administered over a period of 1, 2, 3, 4, 5 or 6 hours. In one embodiment, glufosfamide is administered once every 3 weeks or every 4 weeks (treatment cycle time) at a dose of 4.5 g / m ^{2 over} an infusion time of 1, 4 or 6 hours. In other embodiments, glufosfamide is (a) 1.5 to about 3.0 g / m ² (for 3 consecutive weeks every 3 weeks (Days 1, 2 and 3) for 1 to 6 hours infusion time (B) in an amount ranging from 2.0 to about 3.0 g / m ² ); (b) 1.5 to about 3.0 g / m ² (in case of 1 to 6 hours of injection once per week) From 2.0 to about 3.0 g / m ² ); or (c) 1.5 to about 6.0 g / m ² (sometimes 4.5 ˜about 6.0 g / m ² , optionally about 5.0 g / m ² ).

In one embodiment, the glufosfamide has a glufosfamide concentration or C _max in the patient's plasma of at least 50 microgram / mL, or at least 100 microgram / mL, or at least 150 microgram / mL, or at least 250 microgram / mL. Dosage to reach. In one embodiment, such plasma levels of glufosfamide are about 1.5 to about 4.5 g / m ² (optionally 2.5 to about 4.5 g / m ^{2 over} an infusion time of 1 to 4 hours. This is achieved by administering glufosfamide in an amount in the range of In one embodiment, the amount of glufosfamide administered is 5.0 g / m ² . In one embodiment, glufosfamide is administered such that the area under the concentration time curve (AUC) of glufosfamide is at least 200 megogram-h / mL, or at least 500 megogram-h / mL, or at least 1000 megogram-h / mL. . In one embodiment, such plasma levels of glufosfamide are about 1.5 to about 4.5 g / m ² (optionally 2.5 to about 4.5 g, respectively) over an infusion time of 1 to 4 hours. / M ² ) by administering glufosfamide in an amount in the range.

Glufosfamide is a prodrug that is converted in vivo to the active metabolite isophosphoramide mustard (IPM), and in one embodiment, glufosfamide or other IPM prodrug or IPM itself (which is typically Co-administered with MESNA to avoid adverse side effects as known in the art) with an IPM concentration in the patient's plasma of at least 1 microgram / mL, or at least 3 microgram / mL, or at least 4 microgram / Dosage to reach mL. In one embodiment, such plasma levels of IPM are about 1.5 to about 4.5 g / m ² (optionally 3.5 to about 4.5 g / m ² ), respectively, over a 4 hour infusion time. ² ) achieved by administering glufosfamide in an amount in the range. In one embodiment, glufosfamide or other IPM prodrug or IPM itself has an IPM AUC of at least 10 microgram-h / mL, or at least 25 microgram-h / mL, or at least 35 microgram-h / mL. To be administered. In one embodiment, such plasma levels of IPM are about 1.5 to about 4.5 g / m ² (optionally 2.5 to about 4.5 g, respectively) over an infusion time of 1 to 4 hours. / M ² ) by administering glufosfamide in an amount in the range. In one embodiment, such plasma levels of IPM are achieved by administering glufosfamide at 5.0 g / m ² .

In one embodiment, glufosfamide is administered as described above, and gemcitabine is also administered for 1, 2, 3, 4, 5, 6, 7, 8 or more than 8 treatment cycles, and gemcitabine in each administration cycle Each dose of about 1000 mg / m ² to about 2200 mg / m ² (optionally about 1000 mg / m ² to about 1500 mg / m ² ; and optionally about 1500 mg / m ² ) over an infusion time ranging from about 30 to about 150 minutes. m ² to about 2200 mg / m ² ), including intravenous infusion of gemcitabine. In one embodiment of co-administering glufosfamide and gemcitabine, gemcitabine is administered at weeks 1, 2, 3, 4, 5, 6 and 7 of the treatment cycle for 1, 2, 3, 4 or more than 4 treatment cycles. Here, each cycle is a 7-week cycle. In one embodiment, gemcitabine is administered in the first, second and third weeks of the treatment cycle for 1, 2, 3, 4, 5, 6 or more treatment cycles, wherein each cycle is a 4-week cycle . Gemcitabine can be administered according to the method of the present invention the day before, the next day, or the same day as the administration of glufosfamide. In one embodiment, gemcitabine is administered about 30 minutes to about 4 hours after the administration of glufosfamide on the same day as the administration of glufosfamide.

In one embodiment of co-administering glufosfamide and gemcitabine, gemcitabine is administered such that the gemcitabine concentration in the patient's plasma reaches at least 20 micrograms / mL. In one embodiment, this plasma level of gemcitabine is achieved by administering gemcitabine in an amount of about 1 g / m ^{2 over} an infusion time of about 0.5 hours. Gemcitabine is a prodrug that is converted in vivo to the active metabolite dFdU, and in one embodiment, gemcitabine is administered such that the concentration of dFdU in the patient's plasma reaches at least 30 micrograms / mL. In one embodiment, this plasma level of dUdF is reached by administering gemcitabine in an amount of about 1 g / m ^{2 over} a 0.5 hour infusion time.

  In a second aspect, the present invention provides a method of treating cancer, comprising administering glufosfamide to a subject in need of treatment for advanced and / or metastatic malignancy previously treated with chemotherapy. Including. In one embodiment, the present invention provides a method for the treatment of gemcitabine refractory pancreatic cancer.

  In a third aspect, the present invention relates to an agent having anti-tumor activity, such as bevacizumab (Avastin®), carboplatin, cetuximab (Erbitux®), cisplatin, decarbazine (DTIC), 2-deoxyglucose, doxorubicin (Eg Doxil® and Caelix®), EGFR inhibitors (eg Iressa), erlotinib (Tarceva®), etoposide, exatecan, imatinib mesylate (Gleevec®), irinotecan, methotrexate , Panvac-VF®, pemetrexed (Alimta®), rituximab, rubitecan (Orathecin®), taxanes (eg docetaxel and Taxel), topotecan, vincristine or trastuzumab (Herceptin®) in combination with glufosfamide or in combination with glufosfamide and gemcitabine or in combination with glufosfamide, gemcitabine and erlotinib A method for such treatment is provided.

(Detailed description of the invention)
In one aspect of the invention, glufosfamide and optionally gemcitabine and optionally one or more additional anticancer agents are administered to a subject in need of treatment for cancer in a therapeutically effective amount.

  The antineoplastic agent glufosfamide (β-D-glucosyl-isophosphoramide mustard; gle-IPM) is an alkylating agent that has been investigated for use in the treatment of cancer (US Pat. No. 5,622, incorporated herein by reference). 936, and Niculescu-Duvaz, 2002, Curr Opin Investig Drugs 3: 1527-32. The alkylating moiety (isophosphoramide mustard, IPM) is β-D-glucose glycoside linked and the cellular uptake of glufosfamide is a sodium-dependent transmembrane glucose transporter protein (Briasculis et al., 2000, J Clin Oncol 18: 3535-44) and other glucose transporters described herein. In phase II clinical trials, glufosfamide is a pancreatic cancer patient receiving first-line treatment and a non-small cell lung cancer patient receiving second-line chemotherapy, as well as glioblastoma, breast cancer and colon cancer. In some patients, glufosfamide has been administered (see above, Niculescu-Dubaz, 2002). Glufosfamide is administered intravenously; it is contemplated that other routes of administration may be used in the practice of the present invention, such as intrathecal administration, intratumoral injection, oral administration and other routes.

  In one embodiment of the invention, gemcitabine is co-administered with glufosfamide. Gemcitabine (2'-deoxy-2 ', 2'-difluoro-cytidine, also called 1- (4-amino-2-oxo-1H-pyrimidin-1-yl) -2-desoxy-2,2-difluororibose Is a nucleoside analog that disrupts the process of cell replication. (See US Pat. Nos. 4,808,614 and 5,464,826, each incorporated herein by reference). Gemcitabine HCl (Gemzar ™; Eli Lilly) has been used for the treatment of patients with non-small cell lung cancer and pancreatic cancer. Gemcitabine HCl is routinely formulated as a sterile solution and is administered by intravenous infusion. Other salt forms such as monophosphates, sulfates, malonates, citrates and succinates are readily prepared and can be utilized if desired. It is contemplated that other routes of administration can be used such as intratumoral injection, intrathecal administration and others. As noted above, in certain embodiments of the invention, gemcitabine is co-administered with glufosfamide to treat cancer patients.

  As used herein, a “subject” or “patient” is a mammal in need of treatment for cancer. In general, the subject is a human cancer patient. In some embodiments of the invention, the subject is a non-human mammal used in a model system (eg, a non-human primate, mouse or rat used in drug screening, characterization and evaluation).

  As used herein and well known in the art, “treatment” is a strategy for obtaining beneficial or desired medical results, including clinical results. For the purposes of the present invention, beneficial or desired clinical results include, but are not limited to, alleviation or remission of one or more symptoms, attenuation of the extent of the disease, stabilized (ie, not exacerbated) condition of the disease, disease Prevention of dilation of the disease, slowing or slowing the progression of the disease, ameliorating or alleviating the disease state, and calming (partially or totally unrelated). “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment, or if receiving different treatment. As demonstrated by the present invention, glufosfamide can enable effective treatment of cancer by administration alone or in combination with other anticancer agents.

  As used herein, and as is well known in the art, a “therapeutically effective amount” of a drug is beneficial or desired medical result, clinical result, eg, description of treatment, when administered to a subject with cancer. The amount of chemical that will have the above. A complete therapeutic effect does not necessarily occur with a single dose and may occur only after administration of a series of doses. That is, a therapeutically effective amount may be administered in one or more administrations.

  When drugs are administered as part of the same course, two (or more) drugs are administered to the subject “in combination”. The course of treatment, in this context, is additive, complementary, synergistic, or otherwise in combination, rather than expected for administration of any combination of agents when administered alone. Refers to administration of a combination of drugs that are believed to provide favorable results. The course of treatment can be one day or several days, but for cancer drugs, more often it is extended to weeks and multiple treatment cycles, each of which is from 1 week to 1 month as described below. Continue for more.

  That is, an example of administration in combination is illustrative and not limiting, but according to the method of the present invention is administration of glufosfamide once every 3 weeks for at least 2-8 or more 3 week treatment cycles, Here, the initial administration of glufosfamide is defined as starting on day 1, and in conjunction therewith, administration of gemcitabine for one or more 7 week cycles, during the first, second, third of the seven week cycle , Once every week in week 5, 6 and 7, where gemcitabine administration starts on day 1, day -1 (one day before day 1) or day 2. Another example of combination administration is administration of glufosfamide once every four weeks starting in the first week (day 1) of a four week treatment cycle for 2 to 8 or more 4 week cycles, and 2 to 8 or more of 4 Gemcitabine administration in the first, second and third weeks (Days 1, 8 and 15) of the same 4-week treatment cycle during the week cycle. In one embodiment, gemcitabine is administered about 30 minutes to about 4 hours after administration of glufosfamide on the same day as glufosfamide.

  Various schedules can be used when administering two or more drugs in combination. In one case, for example, but not limited to, the first dose of Drug 1 is administered prior to the administration of Drug 2, and the treatment with Drug 1 is continued throughout the course of the administration of Drug 2; Later, drug 1 is administered for the first time; or drug 1 is administered for the first time simultaneously with the start of another (drug 2) cancer therapy. As used in this regard, “simultaneously” means that the two drugs are administered on the same or consecutive days.

  In principle, certain drugs can be co-formulated and delivered in combination, but combination therapy generally involves the administration of drugs formulated in separate compositions. Similarly, certain drugs can be administered simultaneously, but more frequently (especially in the case of drugs administered by intravenous infusion), drugs are administered on the same or consecutive days, or at different times according to other schedules. For example, in one embodiment of the present invention, glufosfamide is administered once every 4 weeks for 4 hours infusion time starting on the first day of the 4-week cycle, and gemcitabine is administered first in the same 4-week cycle, On days 8 and 15, administered over a 30 minute infusion time, and administration of both drugs is continued for 1 to 8 or more 4 week cycles. On the first day, gemcitabine is administered 30 minutes after the administration of glufosfamide.

  In another embodiment of the invention, glufosfamide is administered as a “single agent”, ie without being combined with another antitumor agent. For example, glufosfamide can be administered to treat advanced and / or metastatic malignancies previously treated with chemotherapy, in which the glufosfamide is used as a single agent, the previously used drug 1 It can be administered in combination with one or more or in combination with other agents that have not yet been used to treat the patient. In another embodiment, the present invention provides a method for treating locally advanced unresectable or metastatic pancreatic adenocarcinoma previously treated with chemotherapy, and in this embodiment, glufosfamide is a single agent. It may be administered as an agent or in combination with other anticancer agents. In another embodiment, the present invention provides a method for treating gemcitabine refractory metastatic pancreatic adenocarcinoma, again in this embodiment glufosfamide is used as a single agent or in combination with gemcitabine or for the treatment of pancreatic cancer. May be administered in combination with another drug used in

  The methods of the present invention may be any cancer, including but not limited to adenocarcinoma; adenoma; basal cell carcinoma, bone cancer, brain cancer, eg primary brain tumor; breast cancer; adrenal, bronchial, colon, bladder, head and neck , Kidney, larynx, nerve tissue, parathyroid, rectal, stomach and thyroid cancer; cervical dysplasia and in situ carcinoma; epidermoid carcinoma; Ewing sarcoma; giant cell tumor; pleomorphic glioblastoma; Hyperplasia; Hyperplastic corneal neuroma; Intestinal ganglion cell tumor; Islet cell carcinoma; Leiomyoma; Leukemia; Liver cancer; Lung cancer; Tumor of both acute and chronic lymphocytes and granulocytes; Lymphoma; Malignant hypercalcemia; malignant melanoma; Marfan syndrome-like constitutional tumor; medullary carcinoma; mucosal neuroma; mycosis fungoides; myeloma; neuroblastoma; osteosarcoma; ovarian cancer and tumor; Cytoma; polycythemia vera; prostate cancer; kidney cells Retinoblastoma; sarcomas such as Kaposi's sarcoma, osteogenic and other sarcomas, rhabdomyosarcomas and soft tissue sarcomas; seminoma; skin cancers such as metastatic and local skin diseases; small cell lung cancer; It can be used for the treatment of both ulcerative and papillary squamous cell carcinoma; veticulum cell carcinoma; and Wilms tumor.

  The methods of the invention are particularly effective in the treatment of certain cancers such as breast cancer, colorectal cancer, bladder cancer, non-Hodgkin lymphoma, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, sarcoma and gastric cancer.

  In one embodiment of the invention, the subject undergoing glufosfamide treatment has breast cancer. Breast cancer is generally treated by various combinations of surgery, radiation therapy, chemotherapy, and hormone therapy. According to the methods of the present invention, glufosfamide is administered alone or in combination with gemcitabine or in combination with other anticancer agents for the treatment of breast cancer.

  In one embodiment of the invention, glufosfamide is administered to a subject in need of treatment for colorectal cancer. In certain embodiments, the patient being treated has colorectal cancer or metastatic colorectal cancer. Colorectal cancer or metastatic colorectal cancer is currently treated by radiation therapy, surgery and / or chemotherapy (eg, administration of 5-fluorouracil). In one embodiment, glufosfamide is administered as a single agent to treat colorectal cancer; in another embodiment, glufosfamide is co-administered with gemcitabine to treat colorectal cancer. In another embodiment, glufosfamide and bevacizumab are administered in combination to a subject in need of treatment for colorectal cancer; optionally gemcitabine or 5-fluorouracil is also co-administered.

  In one embodiment of the invention, glufosfamide is administered as a second-line therapy for non-Hodgkin lymphoma after unsuccessful CHOP (cyclophosphamide, adriamycin, vincristine and prednisone) therapy.

  In one embodiment of the invention, glufosfamide is administered as a single agent or in combination with gemcitabine to a subject in need of treatment for ovarian cancer. In one embodiment, glufosfamide is used to treat ovarian cancer in combination with an optionally approved second option therapy after cis / carboplatin and / or taxane or other first option therapy is unsuccessful. As a second option therapy, as a single agent or in combination with gemcitabine. In another aspect, glufosfamide is administered as a single agent or as a first-line therapy in the treatment of ovarian cancer in combination with gemcitabine and / or cis / carboplatin and / or a taxane and / or other first-line therapy. The

  In one embodiment of the invention, the subject undergoing glufosfamide treatment has pancreatic cancer. Among pancreatic cancers, chemotherapy-refractory pancreatic cancers such as pancreatic cancer refractory to treatment with gemcitabine (eg Aranco et al., 2003, Cancer Invest. 21: 489-96; Kozuch et al., 2001, The Onclogist 6: 488-95; Noble and Goa, 1997, Drugs 54: 447-72N; Stephens et al., 1998, Oncol. Nurs, Forum 25: 87-93; Burris and Stonenio, 1997, Eur. J. Cancer 33: Suppl 1: Suppl. -22; Rothenberg et al., 1996, Ann. Oncol. 7: 347-53) using the methods disclosed herein, eg, glufosfamide alone or in combination with gemcitabine. It can be treated by administration.

  In one embodiment of the invention, glufosfamide is administered as a first or second option therapy to treat PET positive sarcoma. A “PET positive” sarcoma is a sarcoma that can be detected by positron emission tomography. In one embodiment, glufosfamide is administered as a second option therapy in the treatment of sarcoma, alone or in combination with other approved second option therapies, after unsuccessful treatment with doxorubicin or with another first option therapy. The In another embodiment, glufosfamide is administered as a first option therapy in the treatment of sarcoma in combination with doxorubicin or in combination with other approved first option therapies.

  In one embodiment of the present invention, glufosfamide is combined with carboplatin or cisplatin, VP16 and / or taxane, after failure of the first option, alone or with gemcitabine, and optionally with other approved second-option therapies. In combination, it is administered as a second-line therapy for small cell lung cancer (SCLC). In another aspect, glufosfamide is administered alone or in combination with gemcitabine as a first option therapy in the treatment of SCLC, optionally in combination with carboplatin or cisplatin, VP16 and / or taxane or other approved first option therapy .

  As is more apparent from the above, in certain embodiments of the invention, another anticancer agent is co-administered with glufosfamide to treat cancer. For these embodiments, particularly preferred anticancer agents are bevacizumab (Avastin®), carboplatin, cetuximab (Erbitux®), cisplatin (cis-diamine dichloroplatinum (II), a divalent inorganic water-soluble platinum. Containing complex; Go and Adjei, 1999, J Clin Oncol. 17: 409-22), decarbazine (DTIC), 2-deoxyglucose, doxorubicin (eg Doxil® and Caelix®), EGFR inhibition Agents (eg Iressa), erlotinib (Tarceva®), etoposide, exatecan, imatinib mesylate (Gleevec®), irinotecan, methotrexate, Panvac-VF® , Pemetrexed (Alimta®), rituximab, rubitecan (Orathecin®), taxane (eg docetaxel and paclitaxel), topotecan, vincristine and trastuzumab (Herceptin®) Include. In various embodiments of the invention, one or more of these agents are administered to a subject in need of treatment for cancer in combination with glufosfamide or in combination with glufosfamide and gemcitabine. In one embodiment, the cancer is pancreatic cancer. In one example, for illustration and not limitation, cancer can be treated by co-administering glufosfamide and erlotinib (Tarceva ™) and optionally gemcitabine according to the methods of the invention.

  Gemcitabine, bevacizumab, irinotecan, exatecan, pemetrexed, cisplatin and erlotinib and other anticancer agents described herein are comparable to those used clinically, for example, at any therapeutically effective dose. Or at the same dose. Specific dosage regimens (eg, recommended effective doses) for known and approved anti-neoplastic agents are known to physicians and are described, for example, in PHYSICIAN's DESK REFERENCE, 2003, 57th Ed. , Medical Economics Company, Inc. Oradell, N .; J; Goodman & Gilman's THE PHARMACOLOGICAL BASIS OF THERAPEUUTICS "2001, 10th Edition, McGraw-Hill, New York, and / or available from Federal Drug Or a brief description of certain of these other anticancer agents is given below.

  Bevacizumab (Avastin ™; Genentech), when used in the present invention, can be administered at doses comparable to those routinely used clinically for other purposes (eg, Yang, 2003, N. Eng). J. Med., 349: 419-21 and Cobleigh et al., 2003, Semin Oncol.30 (5Suppl16): 117-24). Bevacizumab is an anti-vascular endothelial growth factor (VEGF) monoclonal antibody developed as an anti-angiogenic agent for the treatment of cancers such as colorectal cancer, non-small cell lung cancer, breast cancer and other solid tumors. See Salgaller, 2003, Curr Opin Mol Ther, 5: 657-67 and PCT applications WO96 / 30046 and WO98 / 45331. In one embodiment of the invention, bevacizumab in combination with glufosfamide is administered to treat such cancer.

  Erlotinib can be administered at a dose ranging from about 25 mg to about 200 mg when used in the present invention. In one embodiment, erlotinib is administered at a dose ranging from about 100 mg to about 200 mg. In one embodiment, the dose of erlotinib administered in combination with glufosfamide is about 150 mg. Erlotinib is typically administered at least 1 hour after meals.

  Exatecan mesylate (DX-8951f; Daiichi Pharmaceutical Co.) is a water-soluble analog of the plant alkaloid camptothecin that inhibits topoisomerase I. Exatecan mesylate has been developed as a therapeutic agent for the treatment of non-small cell lung cancer, ovarian, fallopian tube or peritoneal cancer and breast cancer. Various doses and dosages of exatecan mesylate for the treatment of cancer have been reported. See, eg, Verschragen et al., 2004 Cancer Chemother Pharmacol. 53: 1-7; Esteva et al., 2003, Cancer 98: 900-7; Braybrook et al., 2003, Living Cancer, 41: 215-9; Royce et al., 2004 Invest New Drugs. 22: 53-61. In one embodiment of the invention, exatecan mesylate in combination with glufosfamide is administered to treat cancers such as non-small cell lung cancer, ovarian, fallopian tube or peritoneal cancer and breast cancer.

Irinotecan (CPT-11, Camptosar®; Pharmacia & Upjohn) is a semisynthetic derivative of the plant alkaloid camptothecin that inhibits topoisomerase I. Developed as an anti-cancer agent for the treatment of colorectal cancer. For the purposes of the present invention, irinotecan can be administered at a dose comparable to that routinely used clinically. For example, and without limitation, a patient can receive Camptosar® in a 90 minute infusion once every three weeks. The starting dose for most patients can be 350 g / m ² , but the dose may be reduced to 300 g / m ² for patients 70 years and older. Camptosar® can also be administered according to a weekly dosing schedule starting at 125 g / m ² . The dose can be given for about 2-4 weeks, and each course of therapy is repeated every 7 weeks. Rothenberg et al., 1996, J Clin Oncol. 14: 1128-35.

  Pemetrexed (Alimta ™) is an anti-folate that inhibits thymidylate synthase, dihydrofolate reductase, glycinamide ribonucleotide formyltransferase and aminoimidazole carboxamide ribonucleotide formyltransferase. Pemetrexed is active against in vitro pancreatic cancer cell lines and has shown activity in patients with advanced pancreatic cancer. Kindler, 2002, Semin Oncol. 29: 49-53 and Adjei, 2003, Expert Rev Anticancer Ther. 3: 145-56. In one embodiment of the invention, glufosfamide is administered in combination with pemetrexed and optionally gemcitabine to treat pancreatic cancer.

  In another aspect, the present invention has been found to be useful in therapeutic ("therapeutically effective") doses, and according to such schedule or dosing regimen, glufosfamide alone or in combination with other anticancer agents. A method of treatment is provided. Chemotherapy for cancer typically involves multiple “rounds” or “treatment cycles” of administration of one or more drugs, where each cycle is one of one or more times according to a particular schedule. Includes administration of the above drugs. The cycle is generally measured in weeks (not necessarily) and can be, for example, 1, 2, 3, 4, 5, 6, 7 or 8 weeks in duration. In some embodiments of the invention, for example, the treatment cycle is a 3-week cycle, a 4-week cycle, or a 7-week cycle. In each treatment cycle, the drug is at least once and according to a specific schedule, eg daily; once a week; multiple times a week on consecutive or non-consecutive days; once per cycle; multiple times per cycle, eg 3 consecutive The day is administered, such as once every 3 weeks cycle. When more than one agent (eg, 2 agents) is administered to a subject, each can be administered according to its own schedule (eg, weekly; once every 3 weeks, etc.) as described above. Administration of two or more drugs ensures that both drugs are administered (at least in part) on the same day for the convenience of the patient and the treating physician, even if they are administered in different cycles. Alternatively, the drug can be adjusted to be administered (at least in part) on consecutive days.

  In the therapeutic methods of the invention in which glufosfamide and gemcitabine (or other drug) are administered in combination, they can be administered in any order. In certain embodiments, glufosfamide is administered the day before, the next day, or the same day of gemcitabine (or other drug) administration. In certain embodiments, gemcitabine (or other drug) is administered on the same day as glufosfamide, and administration of gemcitabine (or other drug) is initiated before, simultaneously with, or after the administration of glufosfamide. In certain embodiments, gemcitabine (or other drug) is administered (ie, initiated) from about 30 minutes to about 4 hours after administration of glufosfamide. Other dosing schedules can be used as generally described herein, and in more specific cases will be determined by the physician.

Chemotherapeutic drugs are typically administered for 1-8 treatment cycles, or longer cycles (eg, from continuous treatment until no additional therapeutic effect can be achieved). In various embodiments of the invention, glufosfamide is administered by intravenous infusion to patients in need of treatment for cancer for 1, 2, 3, 4, 5, 6, 7, 8, or more than 8 treatment cycles. Each cycle is in the following range:
a) about 1.5-4.5-5.0-6.0-about 8.0 g / m once every 3 or 4 weeks for an infusion time of 1, 2, 3, 4, 5 or 6 hours ² (grams of glufosfamide per patient surface area); or
b) about 1.5-2. 2 infusion times of 1, 2, 3, 4, 5 or 6 hours during 1-3 consecutive days (first, second and third days) every 3 or 4 weeks. 0 to about 3.0 g / m < ² >; or c) about 0.5 to 1.5 to 1.66 to 2.0 to once per week for an infusion time of 1, 2, 3, 4, 5 or 6 hours 2.5 to about 4.0 g / m ² ;
Including injection of glufosfamide.

According to the method of the present invention, and whether glufosfamide is administered alone or in combination with other anticancer agents, glufosfamide has more than 1, 2, 3, 4, 5, 6, 7, 8 or 8 treatment cycles. For at least once per treatment cycle, and for each administration of glufosfamide, glufosfamide is administered to the cancer patient by intravenous infusion in an amount ranging from 1.5 to about 6.0 g / m ² . In one embodiment, the amount of glufosfamide administered ranges from about 1.5 to about 4.5 g / m ² . In one embodiment, the amount of glufosfamide administered is 4.5 g / m ² . In one embodiment, the amount of glufosfamide administered is 4.5 g / m ² . Infusion of glufosfamide can be administered over a period of 1, 2, 3, 4, 5 or 6 hours. In one embodiment, glufosfamide is administered once every 3 or 4 weeks (treatment cycle time) at a dose of 4.5 g / m ^{2 over} an infusion time of 1, 4 or 6 hours.

In one embodiment, the glufosfamide has a glufosfamide concentration or C _max in the patient's plasma of at least 50 microgram / mL, or at least 100 microgram / mL, or at least 150 microgram / mL, or at least 250 microgram / mL. Dosage to reach. In one embodiment, such plasma levels of glufosfamide are administered in a quantity ranging from about 1.5 to 2.5 to about 4.5 g / m ^{2 over} an infusion time of 1 to 4 hours. Is reached. In one embodiment, glufosfamide is administered such that the AUC of glufosfamide is at least 200 megogram-h / mL, or at least 500 megogram-h / mL, or at least 1000 megogram-h / mL. In one embodiment, such plasma levels of glufosfamide are in a range of about 1.5 to 2.5 to about 4.5 g / m ² , respectively, over a 1 to 4 hour infusion time. Reached by administration.

Glufosfamide is a prodrug that is converted in vivo to the active metabolite isophosphoramide mustard (IPM), and in one embodiment, glufosfamide or other IPM prodrug or IPM itself is the IPM in the patient's plasma. The concentration is administered to reach at least 1 microgram / mL, or at least 3 microgram / mL, or at least 4 microgram / mL. In one embodiment, such plasma levels of IPM are administered glufosfamide in amounts ranging from about 1.5 to 3.5 to about 4.5 g / m ² , respectively, over a 4 hour infusion period. Can be reached. In one embodiment, glufosfamide or other IPM prodrug or IPM itself has an IPM AUC of at least 10 microgram-h / mL, or at least 25 microgram-h / mL, or at least 35 microgram-h / mL. To be administered. In one embodiment, such plasma levels of IPM have glufosfamide in an amount ranging from about 1.5 to 2.5 to about 4.5 g / m ² , respectively, over an infusion time of 1 to 4 hours. Reached by administration.

In one embodiment, glufosfamide is administered as described above, and gemcitabine is also administered for more than 1, 2, 3, 4, 5, 6, 7, 8, or 8 treatment cycles, and gemcitabine in each treatment cycle each dose of over injection time ranging from about 30 to about 150 minutes, including intravenous infusion of gemcitabine at a dose ranging from about 1000 mg / ^{m 2} ~ about 1500 mg / ^{m 2} ~ about 2200 mg / ^{m 2.}

  In one embodiment of co-administering glufosfamide and gemcitabine, gemcitabine is at 1, 2, 3, 4, 5, 6 and 7 weeks of the 7 week treatment cycle for 1, 2, 3, 4 or more than 4 treatment cycles And glufosfamide is administered in the amounts and frequencies described above. In one embodiment, gemcitabine is administered in the first, second and third weeks of the treatment cycle for 1, 2, 3, 4, 5, 6 or more treatment cycles, wherein each cycle is a 4 week cycle. Yes, and glufosfamide is administered in the amounts and frequencies described above. For example, and without limitation, gemcitabine can be administered according to the methods of the present invention the day before, the next day, or the same day as the administration of glufosfamide. In one embodiment, gemcitabine is administered about 30 minutes to about 4 hours after the administration of glufosfamide on the same day as the administration of glufosfamide.

In one embodiment of co-administering glufosfamide and gemcitabine, gemcitabine is administered such that the gemcitabine concentration in the patient's plasma reaches at least 20 micrograms / mL. In one embodiment, this plasma level of gemcitabine is achieved by administering gemcitabine in an amount of about 1 g / m ^{2 over} an infusion time of about 0.5 hours. Gemcitabine is a prodrug that is converted in vivo to the active metabolite dFdU, and in one embodiment, gemcitabine is administered such that the concentration of dFdU in the patient's plasma reaches at least 30 micrograms / mL. In one embodiment, this plasma level of dUdF is reached by administering gemcitabine in an amount of about 1 g / m ^{2 over} a 0.5 hour infusion time.

  As is known in the art, treatment with a cancer therapeutic agent can be interrupted and then resumed when toxicity is observed, or at the convenience of the patient, without departing from the scope of the present invention.

  In one embodiment, administration of glufosfamide begins after steps have been taken to reduce serum glucose levels. This method of the invention is beneficial in that the lower the glucose concentration in the blood, the more glufosfamide can enter the cancer cell because glucofosamide and glucose compete for the same glucose transporter on the cancer cell. . The blood glucose concentration is not limited, for example, but for a period of time prior to administration of glufosfamide, for example 8-12-16 hours, by prohibiting the patient's intake of food and glucose-containing nutrients and / or insulin and blood glucose levels. It can be reduced by administering another drug to be reduced.

  In another aspect of the invention, the likelihood of a cancer responding favorably (to be treated) to glufosfamide administration can be determined by measuring the level of glucose transporter in the cancer prior to glufosfamide administration. Since glucose transporters are thought to transport glufosfamide into cancer cells, elevated levels of glucose transporters that transport glufosfamide make cancer cells more sensitive to glufosfamide therapy.

  In this aspect of the invention, the presence or level of a biological agent that functions as a glucose transporter marker is detected by examining a patient or patient tumor sample. A method for measuring glucose transporter levels is described in PCT patent application 2004/081181, which is incorporated herein by reference. In general, the higher the level of glucose transporter, the higher the likelihood that the cancer will respond better to glufosfamide therapy. In one embodiment of the invention, the glucose transporter level measured is the level of GLUT7, GLUT8 or GLUT12. Cancers typically having high levels of these transporters include nasopharyngeal cancer, ovarian cancer, squamous and glandular lung cancer, small cell carcinoma, glioblastoma, chondrosarcoma, esophageal cancer, germ cell tumor and multiple cancers Includes multiple myeloma. That is, in one embodiment, the present invention provides a method for treating cancer in a patient, wherein the method first involves a relatively high level of glucose transporter in the patient, such as, but not limited to, GLUT7, GLUT8 or Measuring the expression of GLUT12. In one embodiment, the level of GLUT1, GLUT2, GLUT3 and / or GLUT4 is measured instead of or in addition to the level of GLUT7, GLUT8 or GLUT12.

  Various tests can be used to measure glucose transporter levels in tumors, such as tests that measure transporter RNA levels (eg, by quantitative PCR) and antibody tests that measure glucose transporter protein directly. These tests as well as the PET scan described below (which also gives a measure of glucose transporter levels) can also be used to monitor the therapeutic effect of glufosfamide administration.

  In one embodiment of the invention, a PET scan is used to examine whether the cancer to be treated may respond to glufosfamide therapy alone or in combination with other agents. In this embodiment, a PET scan is used to determine if the cancer cells are actively taking up the PET scan agent 2-fluoro-deoxyglucose, and if so, the physician will have a successful uptake of the drug by the cancer. Determine that they are more likely to respond better to glufosfamide therapy than less severe cancers.

  Other markers can be used in accordance with the methods of the present invention to determine if a patient is or is likely to respond well to glufosfamide therapy. For example, hypoxia, ie low oxygen concentration, in a tumor typically upregulates glucose transporter levels. Osteopontin is a serum marker indicating the presence of hypoxic cells, for example.

  As another example, serum carbohydrate antigen 19-9 has been reported to be a useful marker for assessing response to gemcitabine therapy in pancreatic cancer (Ziske et al., 2003, Br. J. Cancer 89: 1413). -17). In one embodiment, the response to glufosfamide therapy for cancer administered with glufosfamide as a single agent or in combination with other anticancer agents is characterized by measuring the subject's serum carbohydrate antigen 19-9 levels during treatment. It becomes.

  As another example, the level of cancer antigen-125 (CA-125) can be used for monitoring ovarian cancer. In one embodiment, response to glufosfamide therapy for ovarian cancer administered with glufosfamide as a single agent or in combination with other anticancer agents is monitored by measuring the subject's CA-125 levels during treatment and CA A decrease of -125 indicates a therapeutic effect.

That is, the present invention provides various methods for the treatment of cancer. In one embodiment, glufosfamide is administered intravenously at a dose of 4.5 g / m ² once every 4 weeks for an infusion time of 1 to 4 hours, and this administration is repeated one or more times in combination The cancer is treated by intravenous administration of gemcitabine once a week for 3 weeks in each 4-week treatment cycle at a dose of 1 g / m ² for an infusion time of 0.5 to 1 hour; Without limitation, it can be used to treat pancreatic cancer, ovarian cancer and breast cancer. In another embodiment, the cancer is treated by intravenous administration of glufosfamide at a dose of 4.5 g / m ² once every 3 weeks for an infusion time of 1 to 4 hours and repeating this administration one or more times. This embodiment can be used to treat, for example, but not limited to, small cell lung cancer, ovarian cancer and soft tissue sarcoma. In another embodiment, glufosfamide is administered intravenously at a dose of 1.66 to 2.5 g / m ² over an infusion time of 1 to 4 hours once a week, and this administration is repeated one or more times. Treating cancer; this embodiment can be used to treat, for example, but not limited to, ovarian cancer and soft tissue sarcoma.

  In another embodiment, glufosfamide is administered such that the concentration of glufosfamide in the patient's plasma reaches at least 50 micrograms / mL or 100 micrograms / mL or 150 micrograms / mL, and the administration is performed every 4 weeks. Repeated once or more in combination with gemcitabine in an amount such that the concentration of gemcitabine in the patient's plasma reaches at least 20 micrograms / mL, and this administration is repeated during each 4 week treatment cycle Cancer is treated by repeating once a week for 3 weeks; this embodiment can be used to treat, for example, but not limited to, pancreatic cancer, ovarian cancer and breast cancer. In another embodiment, glufosfamide is administered once every three weeks such that the concentration of glufosfamide in the patient's plasma reaches at least 50 micrograms / mL or 100 micrograms / mL or 150 micrograms / mL; The cancer is then treated by repeating this administration one or more times; this embodiment can be used to treat, for example, but not limited to, small cell lung cancer, ovarian cancer and soft tissue sarcoma. In another embodiment, glufosfamide is administered once a week such that the concentration of glufosfamide in the patient's plasma reaches at least 150 micrograms / mL or 250 micrograms / mL, and the administration is repeated one or more times. This embodiment can be used to treat ovarian cancer and soft tissue sarcoma, for example, without limitation.

  In another embodiment, the glufosfamide is administered such that the AUC of glufosfamide is at least 200 micrograms-h / mL, or at least 500 micrograms-h / mL, or at least 1000 micrograms-h / mL, and the administration is 4 Repeat one or more times per week, in combination with gemcitabine in an amount such that the concentration of gemcitabine in the patient's plasma reaches at least 20 micrograms / mL, and this administration is treated for 4 weeks each Cancer is treated by repeating once a week for 3 weeks in the cycle; this embodiment can be used to treat, for example, but not limited to, pancreatic cancer, ovarian cancer, and breast cancer. In another embodiment, the glufosfamide is administered once every three weeks such that the AUC of glufosfamide is at least 200 megogram-h / mL, or at least 500 megogram-h / mL, or at least 1000 megogram-h / mL. And treating the cancer by repeating this administration one or more times; this embodiment can be used to treat, for example, but not limited to, small cell lung cancer, ovarian cancer and soft tissue sarcoma. In another embodiment, glufosfamide is administered once a week such that the concentration of glufosfamide in the patient's plasma reaches at least 500 microgram-h / mL or at least 1000 microgram-h / mL, and Treat cancer by repeating one or more times; this embodiment can be used to treat, for example, but not limited to, ovarian cancer and soft tissue sarcoma.

  In another embodiment, glufosfamide or other IPM prodrug or IPM itself so that the concentration of IPM in the patient's plasma reaches at least 1 microgram / mL or at least 3 microgram / mL or at least 4 microgram / mL And this administration is repeated at least once every 4 weeks, combined with gemcitabine or other in an amount such that the concentration of dFdU in the patient's plasma reaches at least 30 micrograms / mL. Treating the cancer by administering a dFdU prodrug and repeating this administration once a week for 3 weeks in each 4 week treatment cycle; this embodiment includes, but is not limited to, treating pancreatic cancer, ovarian cancer and breast cancer Can be used for. In another embodiment, once every three weeks, glufosfamide or other so that the concentration of IPM in the patient's plasma reaches at least 1 microgram / mL or at least 3 microgram / mL or at least 4 microgram / mL. To treat cancer by administering one or more IPM prodrugs or IPM itself and repeating this administration one or more times; this embodiment includes, but is not limited to, small cell lung cancer, ovarian cancer and soft tissue sarcoma Can be used. In another embodiment, glufosfamide or other IPM prodrug or IPM itself is administered once a week so that the concentration of IPM in the patient's plasma reaches at least 3-4 micrograms / mL, and Treat cancer by repeating one or more times; this embodiment can be used to treat, for example, but not limited to, ovarian cancer and soft tissue sarcoma.

  In another embodiment, glufosfamide or other IPM prodrug or IPM such that the IPM AUC reaches at least 10 microgram-h / mL or at least 25 microgram-h / mL or at least 35 microgram-h / mL. Gemcitabine in an amount so that the concentration of dFdU in the patient's plasma reaches at least 30 microgram-h / mL in combination with this administration and repeated at least once every 4 weeks. Or other dFdU prodrugs and treating the cancer by repeating this administration one or more times per week for 3 weeks in each 4 week treatment cycle; this embodiment includes but is not limited to pancreatic cancer, ovary Can be used to treat cancer and breast cancer. In another embodiment, glufosfamide such that the IPM AUC reaches at least 10 microgram-h / mL or at least 25 microgram-h / mL or at least 35 microgram-h / mL once every three weeks. Treating cancer by administering other IPM prodrugs or IPM itself and repeating this administration one or more times; this embodiment includes, but is not limited to, small cell lung cancer, ovarian cancer and soft tissue sarcoma Can be used for In another embodiment, glufosfamide or other IPM prodrug or IPM so that the IPM AUC in the patient's plasma reaches at least 25 microgram-h / mL or at least 35 microgram-h / mL once a week. The cancer is treated by administering itself and repeating this administration one or more times; this embodiment can be used to treat, for example, but not limited to, ovarian cancer and soft tissue sarcoma.

  Although the invention has been described in detail in the foregoing sections, the following examples are presented to illustrate certain features of the invention and are not intended to be limiting.

Example 1
(Cell proliferation test)
Mia-PaCa-2, AsPC-1 and H766t cell lines were obtained from American Type Culture Collection (ATCC, Rockville, MD). All cell lines were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum, sodium pyruvate, non-essential amino acids, L-glutamine, vitamins and antibiotics. Cells were maintained at 37 ° C. in a humidified incubator containing 10% CO ₂ . Unless otherwise noted, all chemical reagents were purchased from Sigma Chemical Co. (St. Louis, MO). Gemcitabine is available from Eli Lilly Co. Purchased more and reconstituted in sterile phosphate buffer (PBS) and stored in aliquots at room temperature for in vivo testing and at 20 ° C. for in vitro testing. Glufosfamide was prepared as described in US Pat. No. 5,622,936 and German Patent DE 19534366 and was reconstituted freshly in PBS for each test. Anti-proliferating cell nuclear antigen (PCNA) and anti-CD31 monoclonal antibody were obtained from Dakocytomation Corporation (Carpinteria, Calif.). Cells were harvested from exponential growth medium. Cell number was measured by direct counting in a hemocytometer and cell viability was measured by trypan blue excretion. For cell growth curves, 1 × 10 ⁶ cells were plated in triplicate for each dose level. After 24 hours, the medium was removed and replaced with fresh medium mixed with gemcitabine 1 μg / ml, glufosfamide 10 μg / ml or both. These concentrations are selected based on preliminary tests and are approximately 50% inhibitory concentrations. Viable adherent cells were counted directly with a hemocytometer daily for 3 days.

  Based on preliminary experiments, a concentration of 1 μg / ml gemcitabine was selected to examine its growth inhibitory effect. At this concentration, gemcitabine suppressed the growth of MiaPaCa-2 and H766t cells, but showed only a moderate effect on the growth of AsPC-1 cells (FIG. 1). Glufosfamide at a concentration of 10 μg / ml was less effective than gemcitabine. However, when glufosfamide was added to gemcitabine, additional suppression of cell growth in MiaPaCa-2 and H766t cells was observed, but not in AsPC-1 cells.

(Example 2)
(Measurement of DNA fragmentation by fluorescence activated cell sorting (FACS) analysis)
Quantification of DNA fragmentation (apoptosis) was performed based on the method of Nicoletti et al. (1991). 1 × 10 ⁶ cells were incubated for 24 hours with vehicle, gemcitabine (1 ug / ml), glufosfamide (10 ug / ml) or a combination of both gemcitabine and glufosfamide (1 and 10 μg / ml, respectively). Cells were then collected by gentle trypsinization, washed with PBS, and pelleted by centrifugation. Cells were resuspended in PBS containing 50 μg / ml propidium iodide, 0.1% Triton X-100 and 0.1% sodium citrate. Samples were then stored for 16 hours at 4 ° C. and subjected to FACS analysis after rotary mixing. The relative percentage of cells in the pre-G1 region was then quantified and used as an estimate of cells undergoing apoptosis. The FACS channel was kept constant throughout the experiment based on reference analysis.

  Since cytotoxicity studies have shown that glufosfamide and gemcitabine treatment is most effective against MiaPaCa-2 cells, this combination can be used to induce apoptosis by performing FACS analysis of DNA fragmentation of MiaPaCa-2 cells. It was investigated whether it was induced. Treatment of MiaPaCa-2 cells with glufosfamide 10 μg / ml or gemcitabine 1 μg / ml resulted in induction of apoptosis as measured by increased apoptosis (G1 prefraction; FIG. 2), but gemcitabine had a high effect. . Treatment with both agents resulted in enhanced apoptosis that was additive in nature.

(Example 3)
(Animal model test)
Using MiaPaCa-2 cells that have been stably transfected to express red fluorescent protein (RFP) as previously described (Katz et al., 2003, J. Surg. Res. 113: 151-60). An orthotopic tumor formed. A stock strain of tumor was prepared by injecting MiaPaCa-2RFP cells subcutaneously into the body of nude mice at a concentration of 5 × 10 ⁶ cells / 200 μL. Tumor tissue was removed aseptically and any tissue that was necrotic or suspected to be necrotic or non-RFP tumor was removed. The remaining healthy tumor tissue was then fragmented to a size of about 1 mm ³ . Recipient mice were anesthetized with isoflurane and the surgical area was sterilized with iodine and alcohol. The left upper abdomen of a nude mouse was incised about 1.5 cm in length using surgical scissors. The pancreas was exposed, and then the capsule at the transplant site was peeled off, and then two MiaPaCa-2-RFP tumor fragments were transplanted into the mouse pancreas using 8-0 surgical suture (nylon). The abdomen was closed with 6-0 surgical suture (silk). The entire procedure was performed using a 7X microscope (Olympus) under a HEPA filtered laminar flow hood.

During the course of the study, the primary tumor size of each animal was observed weekly. The primary tumor size was estimated by measuring the short dimension (W) and the long dimension (L) that were orthogonal while using a sliding caliper. Tumor volume as an approximate value was calculated by the formula (W ² × L) × 1/2. At the end of the study, the primary tumor burden was confirmed and metastasis to major organs was assessed by whole body imaging as previously described. In summary, mice were placed in a fluorescent box (Lighttools Research, Encinitas, Calif.) Equipped with a 490 nm fiber optic light source, and images were acquired with a Leica fluorescent stereomicroscope model LZ12 equipped with a mercury lamp and a 50 W power supply. Selective excitation of red fluorescent protein (RFP) was performed through a D425 / 60 bandpass filter and a 470DCXR dichroic mirror. The emitted fluorescence was passed through a long-pass filter GG475 (Chroma Technology, Brattleboro, VT) to a Hamamatsu C5810 type 3-chip cooled color charge-coupled device camera (Hamamatsu Phostronics on Hamamatsu Phostronics). Images were processed for contrast and brightness and analyzed using Image Pro Plus 3.1 software (Media Cybernetics, Siler Spring, Maryland). High resolution images of 1024 x 724 pixels were captured continuously via image output directly on an IBM PC or on a high resolution Sony VCR (SLV-R1000 type; Sony, Tokyo).

In a preliminary experiment, glufosfamide was injected IV at a dose of 3-100 mg / kg / day for 14 days using 6 mice per group. Control mice received saline. Based on the results of this study, doses of 10 and 30 mg / kg / day were selected for the second study. Metastasis frequencies of all groups were analyzed using Fisher's exact test. Comparison of animal survival between each treatment group and untreated controls was analyzed using log rank analysis. Comparison of tumor size was examined by the Kruskal-Wallis test followed by log rank analysis. All values were considered significant when P <0.05. FIG. 3A shows the final tumor volume recorded at necropsy for glufosfamide administered at 0, 3, 10, 30, and 100 mg / kg IV for 14 days, and 300 mg / kg IP gemcitabine (Gem) once weekly for 3 weeks. Treatment started when the tumor reached 100-150 mm ³ . Glufosfamide treatment resulted in a dose-related decrease in tumor volume, and the 100 mg / kg dose was similar in effect to gemcitabine. Furthermore, at doses of 30 mg / kg and higher, a significant increase in time to 50% survival, ie, an effect that was not significantly different from that observed with gemcitabine was observed (FIG. 3B).

(Example 4)
Glufosfamide and Gemcitabine Combination Therapy A combination of glufosfamide and gemcitabine was administered daily for 7 days to female NMR Inu / nu mice carrying tumors derived from H766-T or As-PC-1 human pancreatic cancer cells. Mice received vehicle control (0.9% sodium chloride solution, iv), gemcitabine (ip), glufosfamide (iv), or a combination of gemcitabine / gulphosphamide once a day for 7 consecutive days. The standard dose of gemcitabine used throughout the study was 1.25 mg / kg.

  In the HS766-T tumor model, 10 mg / kg glufosfamide administration resulted in a dramatic delay in the onset of tumor growth compared to gemcitabine alone or the solvent control group. Gemcitabine treatment alone produced only a borderline inhibitory effect on tumor progression. The combined administration of 10 mg / kg glufosfamide and 1.25 mg / kg gemcitabine was statistically significant (P = 0.0099) at the end of the study period (day 42) compared to glufosfamide monotherapy (day 42). ) Brought about a decrease. In the As-PC-1 tumor model, the dosage regimen of glufosfamide and gemcitabine, either alone or in combination, did not have a clear inhibitory effect on tumor growth. These results indicate that the HS766-T tumor model is sensitive to combined administration of glufosfamide and gemcitabine.

(Example 5)
(Glufosfamide and gemcitabine combination therapy)
A combination of glufosfamide and gemcitabine was administered to nude mice bearing tumors derived from MiPaca type 2 human pancreatic cancer cells. Mice were dosed with vehicle control, gemcitabine, glufosfamide or gemcitabine / gluphosphamide combination (10 / group) as shown in Table 1 below. Glufosfamide was administered iv daily for 14 days (groups 2-4 and 7-12). Gemcitabine was administered at 300 mg / kg ip once a week for 3 weeks (Groups 6 and 10-12). Gemcitabine was administered at 150 mg / kg ip twice a week for 3 weeks (Groups 5 and 7-9).

群３及び４は中等度の腫瘍サイズ減少を示した。腫瘍サイズの約５０％減少が群６において観察された。群１０は群６と同等であった。群１１及び１２は群６と比較してより大きい腫瘍サイズ減少を示した（群１２は毒性と考えられた）。この実験においては、１５０ｍｇ／ｋｇにおけるゲムシタビン投与（群５及び７〜９）は動物の急速な死亡をもたらした。グルホスファミド３０ｍｇ／ｋｇとゲムシタビンの組み合わせ（群１１）はゲムシタビン単独（群６）より有意に高値の有効性を示した。図４はグルホスファミドの存在下又は非存在下にゲムシタビン３００ｍｇ／ｋｇを投与した群に関する週毎の原発腫瘍体積を示す。結果によれば、グルホスファミドとゲムシタビンの組み合わせを投与した動物における腫瘍の縮小はゲムシタビン単独の場合よりも有意に高値の有効性を示している。治療の同じ組み合わせはまた生存の有意な向上ももたらした。

Groups 3 and 4 showed moderate tumor size reduction. About 50% reduction in tumor size was observed in group 6. Group 10 was equivalent to Group 6. Groups 11 and 12 showed greater tumor size reduction compared to Group 6 (Group 12 was considered toxic). In this experiment, gemcitabine administration at 150 mg / kg (Groups 5 and 7-9) resulted in rapid death of the animals. The combination of glufosfamide 30 mg / kg and gemcitabine (group 11) showed significantly higher efficacy than gemcitabine alone (group 6). FIG. 4 shows the weekly primary tumor volume for groups administered gemcitabine 300 mg / kg in the presence or absence of glufosfamide. The results show that tumor shrinkage in animals administered the combination of glufosfamide and gemcitabine is significantly higher than that of gemcitabine alone. The same combination of treatments also resulted in a significant improvement in survival.

(Example 6)
(Immunohistochemical features of pancreatic tumors)
In another experiment, tumors were established in groups of 5 nude mice by intrapancreatic injection of 1 × 10 ⁶ Mia-PaCa-2 cells after a midline incision and laparotomy under isoflurane anesthesia and aseptic conditions. Treatment was initiated with glufosfamide IV 30 mg / kg / day for 14 days, gemcitabine IP 300 mg / kg / day (first and eighth days of treatment) or both drugs for 14 days when the tumor size reached approximately 150 mm ³ . PBS was used as a vehicle control. The day after the last glufosfamide dosing, tumors were collected and frozen until analysis.

Frozen tissue sections were fixed and paraffin-embedded 5 μm sections were made and terminal deoxynucleotidyl transferase (Tdt) mediated nick end labeling (using a commercial kit (Promega, Madison, WI) according to the manufacturer's instructions ( TUNEL). Background reactivity was determined by processing slides in the absence of Tdt (negative control). Nuclei were stained with propidium iodide for 10 minutes. Fluorescence bleaching was minimized with an enhancement reagent (Prolong; Molecular Probes, Eugene, OR). Immunofluorescence microscopy was performed with a fluorescence microscope equipped with a narrow bandpass excitation filter. Images were captured using a Nikon camera (Photometrics, Tucson, AZ). DNA fragmentation was detected by localized green fluorescence in the nucleus of apoptotic cells. For quantification of total TUNEL expression, the number of apoptotic events was counted in 10 random 0.159 mm ² fields at a magnification of x100.

To detect PCNA and CD31 immunohistochemically, paraffin-embedded tissues were mounted on positively charged Superfrost slides (Fisher Scientific, Houston, TX) and dried overnight. Sections were deparaffinized in xylene followed by a series of graded alcohol [100%, 95% and 80% ethanol / double distilled water (v / v)] treatment and in PBS (pH 7.5) Rehydrated. To enhance antigen recovery, sections were microwaved for 5 minutes. For detection of CD31, paraffin-embedded tissues were treated with pepsin (Biomeda) at 37 ° C. for 15 minutes and washed with PBS. After exposure to anti-PCNA or anti-CD31 antibody and washing with PBS, positive reactions were visualized by incubating slides with stable 3,3-diaminobenzidine for 10-20 minutes. Sections were washed with distilled water, backstained with Gill's hematoxylin for 30 seconds, and mounted using Universal Mount (Research Genetics). Control samples exposed to the secondary antibody alone showed no specific staining. For MVD quantification, 10 random 0.159 mm ² fields at 100 × magnification were captured on each tumor and microvessels were quantified according to previously described methods (26, 27). . For quantification of PCNA expression, the number of positive cells was counted in 10 random 0.159 mm ² fields at x100 magnification.

  FIG. 5 shows apoptosis induced from TUNEL and PCNA stained sections of tumors from mice treated with saline, glufosfamide (30 mg / kg IV daily for 2 weeks), gemcitabine (300 mg / kg once weekly for 2 weeks) or both drugs. And the proliferation index. Apoptosis occurred at low levels in control mice, but was significantly increased by glufosfamide and increased to a greater extent by gemcitabine. Combination treatment resulted in levels of apoptosis that were significantly higher than either drug alone. PCNA staining was significantly higher than controls in both gemcitabine and glufosfamide, indicating a significant antiproliferative effect. The combined treatment resulted in significantly higher PCNA staining compared to either treatment alone. CD31 staining failed to reveal any effect of either treatment alone or in combination therapy.

(Example 7)
(Glufosfamide and gemcitabine combination therapy in patients with advanced solid tumors and pancreatic adenocarcinoma)
The following examples are presented to illustrate the treatment of cancer with glufosfamide and gemcitabine combination therapy. The safety of glufosfamide in combination with gemcitabine will be evaluated and clarified by conducting clinical trials. The pharmacokinetic parameters when administered in combination with glufosfamide (including the active metabolite isophosphoramide mustard or IPM) and gemcitabine (including the active metabolite dFdU) are also evaluated.

  Conducting multicenter trials and subjecting the subjects to two groups (i) subjects with locally advanced and / or metastatic solid tumors (group I) who have been previously treated or do not have an effective standard treatment available; And (ii) Divide into subjects (group II) with advanced / metastatic pancreatic adenocarcinoma not previously treated with chemotherapy. Assign the subject to a cohort.

The total duration of the study for each subject will be up to 27 weeks, including 3 weeks prior to dosing (screening period). The treatment cycle is 4 weeks, glufosfamide is administered on the first day of each 4-week cycle, and gemcitabine is administered at weeks 1, 8, and 15 of each 4-week cycle. Glufosfamide (1500, 2500, 3500 and 4500 g / m < ² >-1 cohort at each dose level) is administered intravenously over 4 hours on the first day of each cycle once every 4 weeks. A quarter of the dose is administered over the first 30 minutes. The remaining three quarters of the dose are administered over the subsequent 3.5 hours. Gemcitabine (1000 g / m ² ) is administered intravenously over 30 minutes on days 1, 8 and 15 of each 4-week cycle. Gemcitabine infusion begins 30 minutes after the end of glufosfamide infusion on the first day of each cycle. In subjects with stable disease or complete or partial response after 2 cycles, treatment is optionally extended for a period of up to 4 additional 4 week cycles. The dosing schedule is shown in Table 2 below.

バイタルサイン、心電図、臨床検査結果及び有害事象を安全性評価の為に使用する。腫瘍の評価、例えばコンピューター断層撮影（ＣＴ）スキャンはベースライン及び対象が試験中の８週毎に実施する。薬物動態パラメーターは投与後所定の時間間隔で得られたグルホスファミド及びゲムシタビンの血漿中濃度から求める。連続血液試料はグルホスファミド及びその代謝産物であるイソホスホラミド（ＩＰＭ）（サイクル１及び２の第１日のグルホスファミド注入開始後０，０．２５、０．５、１、１．５、２、３、４、４．２５、４．５、５、６、８、１２、１６及び２４時間）及びゲムシタビン及びその代謝産物ｄＦｄＵ（サイクル１及び２の第１日のゲムシタビン注入開始後０、１５、３０、４０、５０分及び１、１．５、３．５、７．５、１１．５及び１９．５時間）の血漿中濃度の測定の為に各対象から収集する。血液試料はグルホスファミド／ＩＰＭ及びゲムシタビン／ｄＦｄＵレベルに関して分析する。有効性及び安全性のデータの分析の為に私用した全ての統計学的試験は両側であり、そして０．０５有意水準で実施し、そして９５％信頼区間を計算する。

Vital signs, ECG, laboratory test results and adverse events are used for safety assessment. Tumor assessments, such as computed tomography (CT) scans, are performed at baseline and every 8 weeks the subject is under study. The pharmacokinetic parameters are determined from plasma concentrations of glufosfamide and gemcitabine obtained at predetermined time intervals after administration. Serial blood samples were glufosfamide and its metabolite isophosphoramide (IPM) (0, 0.25, 0.5, 1, 1.5, 2, 3, 4 after the start of day 1 glufosfamide in cycles 1 and 2). 4.25, 4.5, 5, 6, 8, 12, 16, and 24 hours) and gemcitabine and its metabolite dFdU (0, 15, 30, 40 after start of gemcitabine infusion on day 1 of cycles 1 and 2) , 50 minutes and 1, 1.5, 3.5, 7.5, 11.5 and 19.5 hours) are collected from each subject for measurement of plasma concentrations. Blood samples are analyzed for glufosfamide / IPM and gemcitabine / dFdU levels. All statistical tests used for analysis of efficacy and safety data are two-sided and run at the 0.05 significance level and calculate 95% confidence intervals.

For each subject, the following pharmacokinetics for glufosfamide / IPM and gemcitabine / dFdU: time to maximum concentration (T _max ); maximum peak observed concentration (C _max ); log concentration vs time profile during the terminal phase magnitude of slope of the linear regression _{(K el); ln (2} ) · K calculated half-life as _{el (T 1/2);} concentration time 0 o'clock to the last quantifiable concentration time (LQCT) the area under the curve _{(AUC last),} wherein LQTC the last time the sample was taken with a quantifiable _{concentration;} from 0, which is calculated using the linear trapezoidal rule as AUC _{last +} C LQCT _{/ K el} to infinity Area under the concentration time curve (AUC); clearance (Cl) calculated as dose divided by AUC (glufosfamide and gemcitabine only) An apparent steady state volume of distribution (Vss) calculated as the product of clearance (Cl) and {[AUMC / AUC ² ] − [T _1/2 ]}, where AUMC is the first product of the plasma concentration time curve; The area under rate (glufosfamide and gemcitabine only), and T _I is the injection time; the apparent distribution volume (V _β ) in the post-distribution phase calculated as the ratio of Cl to the terminal elimination rate constant K _el (glufosfamide and gemcitabine Only). Dose adjustment AUC and C _max are calculated for each subject by dividing AUC and C _max by dose. Efficacy results are assessed as determined by response rate, response duration, progression free survival and overall survival of patients with pancreatic cancer (6 and 12 months survival and serum carbohydrate 19-9 changes).

19 patients with pancreas (8), bladder (4) and ovary (1), breast (1), liver (1), stomach / duodenum (2), kidney (1) cancer and sarcoma (1) participated I let you. Two DLT (dose limiting toxicity), i.e. 2500 g / ^m Grade 3 in ² Fatigue and 4500 g / ^m thrombocytopenia grade 4 in ² is going. Both cohorts expanded. DLT did not occur in either the 1500 and 3500 g / m ² cohorts. Phase 1 data show that glufosfamide at a dose of 4500 g / m ² can be safely administered in combination with gemcitabine. Five patients have completed all six cycles, and another patient is ongoing. Reasons for discontinuation were progressive disease (11), clinical deterioration (1), adverse events (AE) (1) and death (1). Grade 3/4 neutropenia occurred in 7 patients (5 during cycle 1) and grade 3/4 thrombocytopenia occurred in 5 patients (2 during cycle 1). CrCL decreased to 60 mL / min in one patient. One unconfirmed partial response was reported; 9 out of 17 evaluable patients (53%) evaluated for response at 8 weeks had stable disease (SD), 1 patient completed 8 cycles of therapy Ovarian cancer patients; one was a breast cancer patient who completed 10 cycles of therapy; and 4 were pancreatic cancer patients who completed at least 7 cycles of therapy.

Pharmacokinetic analysis does not suggest any interaction between glufosfamide and gemcitabine. As shown in FIGS. 6 and 7, glufosfamide and IPM show linear pharmacokinetics at 1500-4500 g / m ² with a half-life of about 2-3 hours; the pharmacokinetics of glufosfamide and IPM are cycle independent It is sex. Glufosfamide has a large Vd _ss (about 9-15 L / m ² ) and low clearance (80-90 mL / min / m ² ). Furthermore, as shown in FIGS. 8 and 9, glufosfamide does not affect the pharmacokinetics of gemcitabine and dFdU. Figures 8 and 9 show the plasma concentration vs time profiles of gemcitabine and dFdU on days 1 (ie in the presence of glufosfamide at all doses) and on day 8 (in the absence of glufosfamide) of cycles 1 and 2. The absence of a significant difference in the plasma concentration profile between days 1 and 8 indicates that there is no effect of glufosfamide (at all doses because all doses of glufosfamide data were pooled).

(Example 8)
(Glufosfamide therapy for gemcitabine-refractory metastatic pancreatic adenocarcinoma)
The following examples are presented to illustrate the treatment of gemcitabine refractory metastatic pancreatic adenocarcinoma with glufosfamide therapy. Assessing the safety and efficacy of glufosfamide in subjects with gemcitabine-refractory metastatic pancreatic adenocarcinoma as measured by overall survival compared to best supported care by conducting a multicenter randomized open-label trial To do.

Subjects are divided into two groups of 150 subjects per treatment group: (i) glufosfamide treatment group (Group I) and (ii) best supportive care (BSC, Group II). Group I subjects receive glufosfamide for a period of up to 51 weeks. Glufosfamide is administered intravenously at 4500 g / m ² for up to 17 doses once every 3 weeks for 6 hours. A quarter of the dose is infused over 30 minutes and the remainder over the next 5.5 hours. Subjects may receive palliative radiation therapy, except within ± 48 hours of glufosfamide administration. To assess the efficacy of glufosfamide treatment, Group II subjects will not receive any drug therapy with anti-tumor effects, such as chemotherapy or other systemic cytotoxicity / cytostatic agent-mediated therapy. Other appropriate support measures and concomitant medications that do not have anti-tumor effects, such as analgesics, antibiotics, infusions, hematopoietic colony-stimulating factor (as a therapy, not prophylactic), erythropoietin, acetate for promoting appetite Guest rolls are administered as appropriate. Group I subjects are also provided with best supportive care. The dosing schedule is shown in Table 3 below.

腫瘍評価はベースライン時及び最初の２４週間は６週毎、そしてその後は疾患の進行が報告されるまで９週毎に実施する。薬物動態試料はサイクル１及び２の間に群Ｉの対象から採取する。グルホスファミド／ＩＰＭの血漿中濃度用の血液試料は後述する時間において、サイクル１及び２の第１日に群Ｉの対象から以下の時点、即ち投与前及びグルホスファミド注入終了直前に収集する。追加的な薬物動態パラメーターは群Ｉの２４対象のサブセットに関して測定する（グルホスファミド／ＩＰＭに関するＡＵＣ、Ｃ_ｍａｘ及びＴ_１／２）。血液試料はこれ等のサブセットからサイクル１及び２の第１日に、以下の時間、即ち投与前、グルホスファミド注入開始後０．５（注入速度変更直前）、１、３、６（グルホスファミド注入終了直前）、６．２５，６．５、７、８、１０、１６、２４時間に収集する。

Tumor assessment is performed at baseline and every 6 weeks for the first 24 weeks, and thereafter every 9 weeks until disease progression is reported. Pharmacokinetic samples are taken from Group I subjects during cycles 1 and 2. Blood samples for plasma concentrations of glufosfamide / IPM are collected at the following times from Group I subjects on the first day of cycles 1 and 2 at the following times: pre-dose and just before the end of glufosfamide infusion. Additional pharmacokinetic parameters are measured for a subset of 24 subjects in Group I (AUC, C _max and T _{1/2 for} glufosfamide / IPM). Blood samples are taken from these subsets on the first day of cycles 1 and 2 at the following times: before administration, 0.5 (just before the infusion rate change), 1, 3, 6 (just before the end of glufosfamide infusion) ), 6.25, 6.5, 7, 8, 10, 16, 24 hours.

  Pharmacokinetic parameters for glufosfamide / IPM (described in Example 7 above) (Day 1 of cycles 1 and 2) are calculated for each subject in the 24 subject subset. Effectiveness results are response rate (full response and partial response), response duration, progression free survival, 6 and 12 month survival, change in VAS pain score, and Assessment is based on serum carbohydrate 19-9 response. The primary efficacy hypothesis examined by the trial is that glufosfamide treated subjects with gemcitabine-refractory metastatic pancreatic adenocarcinoma have improved overall survival when compared to best supported care.

Example 9
(Small cell lung cancer (SCLC); extensive recurrence-sensitive SCLC; ovarian cancer; glufosfamide therapy for soft and advanced soft tissue sarcomas)
Ovarian cancer, small cell lung cancer, widespread relapse sensitive SCLC, of glufosfamide over soft tissue sarcomas and progression soft tissue sarcoma 1 hour infusion time in a single dose to 4500 g / ^{m 2} dose or 5000 g / ^{m 2} dose every 3 weeks Treat according to the method of the invention by intravenous administration.

  In one embodiment, the patient has small cell lung cancer and has previously received ICE (ifosfamide / carboplatin / etoposide) or ICE-V (ifosfamide / carboplatin / etoposide / vincristine) therapy. In one embodiment, the patient has small cell lung cancer and is treated with glufosfamide concurrently (ie in combination) with ICE or ICE-V therapy.

Optionally, ovarian cancer patients treated by intravenous administration of glufosfamide over one hour infusion period at a dose of ^{1600g / m 2, 2500g / m} 2 and 5000 g / ^{m 2} is administered once a week.

  Although the invention has been described with reference to specific embodiments thereof, it will be appreciated by those skilled in the art that various modifications can be made without departing from the scope of the invention, and equivalents may be substituted. it can. In addition, many modifications may be made to adapt a particular situation, material, material composition, process, process step to achieve the benefits afforded by the present invention without departing from the scope of the present invention. Such modifications are intended to fall within the scope of the claims appended hereto.

  All publications and patent documents cited herein are hereby incorporated by reference as if such publications or documents were specifically and individually indicated to be incorporated herein by reference. Citation of publications and patent documents is not intended as a teaching that any such document is pertinent prior art, and does not constitute any acceptance as to its content or date.

FIG. 1 shows the effects of glufosfamide and gemcitabine in a proliferation test. MiaPaCa-2 (A), Hst-766 (B) and AsPC-1 (C) pancreatic cells were grown on the medium for 3 days as described in the Examples. Glufosfamide and gemcitabine were added to the media either alone or together at the indicated concentrations, and cell numbers were measured daily using a hemocytometer. FIG. 2 shows the analysis of DNA fragmentation by FACS. 1 × 10 ⁶ cells were incubated with vehicle (A), gemcitabine (B), glufosfamide (C) or a combination of both drugs (D) for 24 hours at 10 μg / ml with addition of propidium iodide. FIG. 3 shows the final tumor volume (A) and 50% survival (B) recorded in the MiaPaCa-2-RFP pancreatic cancer model. Glufosfamide was administered IV daily for 14 days. ^* Means P <0.05 compared to control. FIG. 4 shows the primary tumor volume recorded in the MiaPaCa-2-RFP pancreatic cancer model. Gemcitabine (IP, once a week for 3 weeks) and glufosfamide (IV daily for 14 days) each significantly reduced the tumor volume measured on day 26, whereas the combination of 30 mg / kg and gemcitabine resulted in a significantly higher decrease. Brought. ^* Means P <0.05 compared to control; ^** means P <0.01 compared to control; # means P <0.05 compared to gemcitabine alone. To do. FIG. 5 shows the apoptosis index and proliferation index measured by TUNEL and PCNA staining. Tumors were removed at the end of treatment and processed for IHC. ^* Means P <0.05 compared to control; ^** means P <0.01 compared to control; ^*** means P <0.001 compared to control ; # Means P <0.05 compared to gemcitabine. FIG. 6 shows plasma of glufosfamide at 0, 4, 8, 12, 16, 20 and 24 hours after the start of glufosfamide infusion on day 1 of the treatment cycle at various doses of glufosfamide (1500, 2500, 3500 and 4500 mg / m ² ). Medium concentration (μg / mL) is shown. FIG. 7 shows the metabolism of glufosfamide at 0, 4, 8, 12, 16, 20 and 24 hours after the start of glufosfamide infusion on day 1 of the treatment cycle at various doses of glufosfamide (1500, 2500, 3500 and 4500 mg / m ² ). The plasma concentration (μg / mL) of the product isophosphoramide mustard (IPM) is shown. FIG. 8 shows 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 and 3. after the start of gemcitabine infusion on days 1 and 8 of the treatment cycle at gemcitabine 1000 mg / m ² . The plasma concentration of gemcitabine at 5 hours (μg / mL) is shown. FIG. 9 shows the plasma concentration (μg of gemcitabine metabolite dFdU at 0, 4, 8, 12, 16, 20 and 24 hours after the start of gemcitabine infusion on days 1 and 8 of the treatment cycle at gemcitabine 1000 mg / m ² . / ML).

Claims (24)

A method of treating cancer comprising administering glufosfamide to a cancer patient by intravenous infusion in an amount ranging from 1.5 to about 6.0 g / m ² with an infusion time ranging from 1 to 6 hours. Wherein the administering step is performed at least once every 4 weeks and repeated at least once. Wherein said amount of glufosfamide to be administered in the administering step is 4.5 g / m ^2, and the method of claim 1, wherein implementing at least once said administering step every 3 weeks. The administration said amount of glufosfamide administered in step is 1.5~3.0g / m ^2, and the method of claim 1, wherein implementing at least once a week the administering step. A method of treating cancer, comprising administering to a cancer patient an amount sufficient for glufosfamide to produce a concentration of glufosfamide in the patient's plasma of at least 50 micrograms / mL. . The method of claim 4, wherein the glufosfamide concentration is at least 100 micrograms / mL. The method of claim 4, wherein the glufosfamide concentration is at least 150 micrograms / mL. A method of treating cancer, comprising administering a sufficient amount of glufosfamide to a cancer patient to provide at least 200 microgram-h / mL glufosfamide AUC. 8. The method of claim 7, wherein the glufosfamide AUC is at least 500 microgram-h / mL. 8. The method of claim 7, wherein the glufosfamide AUC is at least 1000 microgram-h / mL. A method of treating cancer, wherein the method produces a prodrug of glufosfamide or isophosphoramide mustard (IPM) or IPM itself to a cancer patient with an IPM concentration in the patient's plasma of at least 1 microgram / mL. Administering a sufficient amount to effect. 8. The method of claim 7, wherein the IPM concentration is at least 3 micrograms / mL. 8. The method of claim 7, wherein the IPM concentration is at least 4 micrograms / mL. A method of treating cancer, wherein the method comprises glufosfamide or another prodrug of isophosphoramide mustard (IPM) or IPM itself in the patient's plasma at least 10 microgram-h / mL for the patient. Administering a sufficient amount to produce an IPM AUC. 14. The method of claim 13, wherein the IPM AUC is at least 25 microgram-h / mL. 14. The method of claim 13, wherein the IPM AUC is at least 35 microgram-h / mL. Gemcitabine is also administered in infusion time ranging from about 30 to about 150 minutes by intravenous infusion, and any claim 1 to 15 each gemcitabine administration is an amount ranging from about 1000mg ^{/ m 2} ~2200mg ^/ m 2 The method of crab. Gemcitabine is also administered by intravenous infusion with infusion times ranging from about 30 to about 150 minutes, and each gemcitabine administration is in an amount sufficient to result in a concentration of gemcitabine in the patient's plasma of at least 20 micrograms / mL. A method according to any of claims 1-15. Gemcitabine is also administered by infusion in an infusion time range of about 30 to about 150 minutes, and each gemcitabine dose is in an amount sufficient to result in a dFdU concentration in the patient's plasma of at least 30 micrograms / mL. A method according to any of claims 1-15. Progressive and / or metastatic malignancy that has not been previously treated or has been previously treated by surgery, radiation or chemotherapy; breast cancer; colorectal cancer; metastatic colorectal cancer; Ovarian cancer; pancreatic cancer; chemotherapy-refractory pancreatic cancer; gemcitabine-refractory pancreatic cancer; sarcoma; PET-positive sarcoma; and small cell lung cancer (SCLC) selected from the group of cancers 18. The method according to any one of 18. Bevacizumab (Avastin®), carboplatin, cetuximab (Erbitux®), cisplatin, decarbazine (DTIC), 2-deoxyglucose, doxorubicin (eg Doxil® and Caelix®), EGFR inhibition Agents (eg Iressa), erlotinib (Tarceva®), etoposide, exatecan, imatinib mesylate (Gleevec®), irinotecan, methotrexate, Panvac-VF®, pemetrexed® (Alimta® trademark) Rituximab, rubitecan (Orathecin®), taxanes (eg docetaxel and paclitaxel), topotecan, vincristine and trastuzumab ( erceptin agent having anti-tumor activity selected from (R)) the group consisting of also a method according to any one of claims 1 to 19 is administered to said patient. 21. The patient is selected for initial or continued treatment with glufosfamide based on tests performed to determine or estimate glucose transporter levels in cancer cells or tissues of the patient's cancer. The method in any one of. 22. The method of claim 21, wherein the test measures the level of a glucose transporter selected from the group consisting of GLUT1, GLUT2, GLUT3, GLUT4, GLUT7, GLUT8, or GLUT12. The method of claim 21, wherein the test is a PET scan. 24. The method of claim 21, wherein the test determines or estimates a level of hypoxia in the patient's cancer tissue.

Images