Patent Application
20080025984
This invention provides a method for treating tumor in a subject comprising administering to the subject an effective amount of: (1) VNP40101M, or its equivalent; and (2) a nucleoside, or a nucleoside analog. This invention also provides a method for inhibiting tumor cell growth comprising contacting the tumor cell with effective amounts of: (1) VNP40101M, or its equivalent; and (2) a nucleoside, or a nucleoside analog.
VNP40101M is a representative form of SHPs. As used herein, VNP40101M covers itself, its salt or other prodrug forms which produce same or similar effective in vivo. Prodrug is an inactive precursor which will be converted into its active form by normal metabolic processes.
VNP40101M and its equivalents are known in the art. See e.g. U.S. Pat. No. 6,855,695 B2, issued Feb. 15, 2005. Other SHPs can also be used in the invention.
The present invention relates to the treatment of cancer, comprising administering to a subject in need thereof an effective amount of VNP40101M in combination with a nucleoside.
These agents may be administered concurrently or sequentially.
In an embodiment, the subjects are mammals. In a further embodiment, the subjects are human.
As used herein, the nucleoside analog is defined as analog which produces substantially the same effect as the nucleoside itself. Nucleoside analogs are capable of inhibiting DNA synthesis or incorporating into DNA such as azacitidine, cladribine, decitabine. gemcitabine, mercaptopurine, thioguanine, fludarabine, clofarabine, troxacitabine, and pentostatin are useful to combine with VNP40101M to treat malignancies.
Cytarabine, a cell-cycle specific antimetabolite, is the most effective drug in the treatment of AML. Cytarabine is phosphorylated intracellularly and incorporated into DNA. By inhibiting DNA polymerases and DNA synthesis, cytarabine is predicted to inhibit DNA repair and enhance the cytotoxicity of VNP40101M.
Other cancer therapies such as radiation or other chemotherapeutics which include, but are not limited to, antimetabolites, etoposide, doxorubicin, taxol, vincristine, cyclophosphamide, mitomycin C, topoisomerase I and topoisomerase II inhibitors (adriamycin, topotecan, campothecin and irinotecan), platinum containing compounds, (cisplatin, carboplatin), tipifarnib (R115777), SCH66336, erlotinib, gefitinib, and gemtuzumab ozogamicin, may be used with VNP40101M or its equivalent. VNP40101M or its equivalent provides synergistic effects when used in combination with these therapies.
An aspect of the present invention relates to the treatment of cancer, comprising administering to a patient in need thereof an effective amount of VNP40101M in combination with at least one therapeutic agent. In an embodiment, the agent is a nucleoside or a nucleoside analog.
The treatment of solid malignant tumors, leukemia, and lymphomas is a preferred embodiment of the present invention. In a further embodiment, the cancer is acute myelogenous leukemia (AML).
The amount of VNP40101M and nucleoside or its analog used according to the present invention is an effective amount for treating cancer. In general, a therapeutically effective amount of the VNP40101M according to the present invention usually ranges from less than about 0.05 mg/kg to about 100 mg/kg of body weight of the patient to be treated, or considerably more, depending upon the compound used, the tumor type to be treated, the ability of the active compound to localize in the tissue to be treated, the route of administration and the pharmacokinetics of the compound in the patient.
VNP40101M is preferably administered in amounts ranging from about 0.5 mg/kg (17.5 mg/m2) to about 50 mg/kg (1750 mg/M2) or more at one time. Nucleoside or its analog is preferably administered in amounts ranging from about 0.1 mg/kg (3.5 mg/m2) to about 150 mg/kg (5250 mg/m2) or more at one time; preferably is given from 1 mg/kg to 100 mg/kg. The duration of treatment may be for one or more days or may last for several months or considerably longer (years) depending upon the disease state treated.
In a more preferred embodiment, VNP40101M is given to the patient at doses of 1 mg/kg to 20 mg/kg and the nucleoside at doses of 10 mg/kg to 80 mg/kg. In another preferred embodiment, the nucleoside is cytarabine (AraC) and the dose ranges from 20 mg/kg to 60 mg/kg. In a still further embodiment the does of VNP40101M is between 100 and 1000 mg/m2 and the dose of AraC is between 500 and 5000 mg/m2.
This invention provides a composition comprising an amount of VNP40101M or its equivalent which produces synergistic effects when used in combination with a nucleoside or its analog in treating tumor.
This invention provides a composition comprising an amount of nucleoside which produces synergistic effects when used in combination with a VNP40101M in treating tumor.
As demonstrated in the below examples, this effective amount of either VNP40101M, its equivalent or nucleoside (its analog), can be routinely determined. These combinations of agents may be delivered intravenously, subcutaneously, intramuscularly, intraperitoneally or orally. Other routes of administration such as inhalation may also be used.
This invention will be better understood from the examples which follow. However, one skilled in the art will readily appreciate that the specific methods and results discussed are merely illustrative of the invention as described more fully in the claims which follow thereafter.
The cytotoxicity of the combination of VNP40101M and Arac on L1210 leukemia was examined using a cell viability assay. Cells were exposed to VNP40101M, alone or in combination with various concentrations of AraC. After 72 hours, the remaining viable cells were quantified by measuring mitochondrial oxidoreductase activity. Concentrations of AraC and VNP40101M used were between 0.75 and 6 uM. Dose-effect analyses (combination index) showing combination effects of VNP40101M and AraC were analyzed. Combination indices of 0.75-0.8 and 0.1-0.3 indicate moderate synergism and strong synergism, respectively (Chou and Talalay, Adv. Enz. Regul., 22, 27-55, 1984). Cytotoxic effect of VNP40101M was evaluated in the presence of AraC on L1210 leukemia and table 1 shows that CLOETAZINE worked synergistically with AraC on leukemia.
The cytotoxicity of the combination of VNP40101M and nucleoside analogs on tumor cell lines is examined using a cell viability assay. Several leukemia cell lines (L1210 and HL-60) and lymphoma cell lines (Raji and Namalwa) are exposed to VNP40101M, alone or in combination with various concentrations of Fludarabine. After 72 hours, the remaining viable cells were quantified by measuring mitochondrial oxidoreductase activity. Concentrations of Fludarabine used are between 0.1 to 100 uM; concentrations of VNP40101M used are between 0.1 to 100 uM. Dose-effect analyses (combination index) showing combination effects of VNP40101M and Fludarabine are analyzed. Combination indices of 0.75-0.8 and 0.1-0.3 indicate moderate synergism and strong synergism, respectively.
Eighty Female Balb/c×DBA/2 (CD2F1) mice were inoculated intraperitoneally (ip) on day 0 with 1×106 L1210 cells in 0.2 mL phosphate-buffered-saline (PBS). The mice were randomly divided into 6 groups; each group consisted of 10 mice. The animals were untreated or treated with a single bolus dose of VNP40101M at 5, or 10 mg/kg, ip, on day 1 with or without AraC (50 mg/kg, ip) on days 1, 3, 5, 7, and 9. Daring the experiments, mice were observed daily for survival. It was determined that mice that survive for more than 60 days after inoculation of L1210 cells might be regarded as long-term survivors. Kaplan-Meier plots were generated, and survival time of animals was analyzed using student T-test. Significance was defined as P<0.05.
Table 2 shows that intraperitoneal inoculation of mice with 1×106 L1210 leukemia cells resulted in a rapid development of ascites followed by death of all animals in untreated control within 18 days and in AraC treatment control within 30 day. VNP40101M treatment at single doses of 5 and 10 mg/kg increased long-term survivors to 50% and 90%, respectively. Therapeutic efficacies or combinational treatments were superior to single agents. VNP40101M, at 5 and 10 mg/kg, plus AraC treatments increased long-term survivors to 80% and 100%, respectively. Our results suggest that VNP40101M significantly enhanced anti-tumor effect of AraC.
Eighty Female Balb/c×DBA/2 (CD2F1) mice are inoculated intraperitoneally (ip) on day 0 with 3×106 L1210 cells in 0.2 mL phosphate-buffered-saline (PBS). The mice are randomly divided into 6 groups; each group consists of 10 mice. The animals are untreated or treated with a single bolus dose of VNP40101M at 5, or 10 mg/kg, ip, on day 1 with or without fludarabine (70 mg/kg, ip) on multiple days between days 1 to 9. During the experiments, mice are observed daily for survival. It is determined that mice that survive for more than 60 days after inoculation of L1210 cells might be regarded as long-term survivors. Kaplan-Meier plots are generated, and survival time of animals is analyzed using student T-test. Significance is defined as P<0.05.
Table 3 shows that intraperitoneal inoculation of mice with 3×106 L1210 leukemia cells resulted in a rapid development of ascites followed by death of all animals in untreated control within 18 days and in fludarabine treatment control within 30 day. VNP40101M treatment at single doses 10 mg/kg increased long-term survivors to 40%. Therapeutic efficacy of combinational treatment was superior to single agents. VNP40101M, at 10 mg/kg plus fludarabine treatments increased long-tern survivors to 90%. Our results suggest that VNP40101M significantly enhanced anti-tumor effect of fludarabine.
AraC is administered by IV contiguous infusion at a dose of 0.5 to 4.0 gm/m2/day for one to six days. Preferably, AraC is administered at a dose of 1.0 to 2.0 gm/m2/day for three to four days. VNP40101M is administered over 15-30 minutes on day 1 to day 3 after AraC infusion. The dose of VNP40101M is between 200 to 700 mg/m2. Preferably, VNP40101M is administered one day after AraC infusion and the dose is between 450 to 650 mg/m2. Multiple cycles of treatment can be repeated.
Clofarabine is administered by IV at a dose of 10 to 50 mg/m2/day for one to six days. Preferably, clofarabine is administered at a dose of 30 to 40 mg/m2/day for five consecutive days. VNP40101M is administered over 15-60 minutes as a single bolus injection before, during, or after clofarabine administration. The dose of VNP40101M is between 200 and 700 mg/m2. Preferably, VNP40101M dose is between 450 and 650 mg/m2. Multiple cycles of treatment can be repeated.
Tipifarnib is administered orally at 200 to 800 mg/dose, one to three doses a day for up to 28 days. Preferably, tipifarnib is administered at 600 mg/dose, two doses per day, for 28 consecutive days. VNP40101M is administered over 15-60 minutes as a single bolus injection before, during, or after administration of tipifarnib. The dose of VNP40101M is between 200 to 700 mg/m2. Preferably, VNP40101M is between 450 and 650 mg/m2. Multiple cycles of treatment can be repeated.
This application claims benefit of U.S. Ser. No. 60/556,565, filed Mar. 26, 2004. The content of this preceding application is hereby incorporated in its entirety by reference into this application.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/US05/10152 | 3/25/2005 | WO | 00 | 9/15/2006 |
| Number | Date | Country | |
|---|---|---|---|
| 60556565 | Mar 2004 | US |
