o-(chloroacetylcarbamoyl)fumagillol and Uterine-Neoplasms

Uterine carcinosarcoma is a highly aggressive gynecological neoplasm that responds poorly to conventional chemotherapy and radiotherapy. Metronomic chemotherapy is accepted as a new approach for cancer treatment, and its underlying mechanism seems to involve the suppression of angiogenesis. However, the efficacy of metronomic and anti-angiogenic therapies against uterine carcinosarcoma is unknown. The anti-angiogenic effect of doxifluridine was assessed in vitro using human umbilical vein endothelial cells (HUVEC) co-cultured with FU-MMT-1 human uterine carcinosarcoma cells. The antitumor and anti-angiogenic effects of metronomic doxifluridine (delivered via oral gavage) in combination with TNP-470 were evaluated in vivo. Tumor vascularity was assessed by contrast-enhanced color Doppler ultrasound, laser Doppler and microvessel density staining. Doxifluridine suppressed tube formation of HUVEC and vascular endothelial growth factor production by FU-MMT-1 cells. Metronomic doxifluridine alone significantly suppressed tumor growth compared with the untreated (control) group, while metronomic doxifluridine in combination with TNP-470 significantly inhibited tumor growth compared with each treatment alone. A significant reduction of intratumoral vascularity was observed in FU-MMT-1 xenografts following treatment with metronomic doxifluridine in combination with TNP-470, as compared with each treatment alone. Intestinal bleeding was only observed when the maximum tolerated dose of doxifluridine was administered in combination with TNP-470. Metronomic doxifluridine chemotherapy in combination with TNP-470 might be effective for uterine carcinosarcoma without marked toxicity, possibly acting via its potent anti-angiogenic effects. Clinical studies are needed to evaluate the safety and efficacy of this treatment in humans.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Combined Chemotherapy Protocols; Carcinosarcoma; Cell Line, Tumor; Cyclohexanes; Endothelial Cells; Female; Floxuridine; Humans; Mice; Mice, Inbred BALB C; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Thrombospondin 1; Thymidine Phosphorylase; Uterine Neoplasms; Vascular Endothelial Growth Factor A; Xenograft Model Antitumor Assays

The purpose of the present study was to develop a new method of chemoembolization to improve the therapeutic effectiveness and safety profile of cancer treatment. A chemoembolization approach was designed for human solid tumors using resorbable calcium-phosphate ceramic microspheres loaded with an agent anti-angiogenic to tumor vasculature in vivo. The human uterine sarcoma cell line FU-MMT-3 was used in this study because this tumor is aggressive and also exhibits a poor response to radiotherapy or any chemotherapy currently used. The calcium-phosphate ceramic microspheres loaded with TNP-470, an anti-angiogenic agent, were injected into FU-MMT-3 xenografts in nude mice three times per week for 8 weeks. The treatment using TNP-470-loaded microspheres suppressed tumor growth, compared to treatment with TNP-470 alone, microspheres alone, and the control. The mean tumor weight after treatment using TNP-470-loaded microspheres was significantly lower than that after treatment with microspheres alone. These ceramic microspheres were remarkably embolized in tumor microvessels as well as in the feeding arteries and a significant reduction of intratumoral vascularity was also demonstrated following treatment with TNP-470-loaded microspheres. Severe loss of body weight was not observed in any mice treated with the TNP-470-loaded microspheres, compared to treatment with TNP-470 alone. These results suggest that targeting tumor vasculature in human uterine sarcoma using calcium-phosphate microspheres might be more effective and safer than the treatment that employs anti-angiogenic agent alone. This new chemoembolization method incorporating an anti-angiogenic agent may contribute to the effective treatment of locally advanced or recurrent solid tumors.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Calcium Phosphates; Cell Line, Tumor; Ceramics; Cyclohexanes; Female; Humans; Mice; Mice, Nude; Microspheres; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Uterine Neoplasms; Xenograft Model Antitumor Assays

Microvascular endothelial cells, which are recruited by tumors, have become an important target in cancer therapy. This study firstly examined the antitumor effect of angiogenesis inhibitor combined with ultrasound (US) irradiation for human cancer in vivo and evaluated its vascularity using color Doppler US in real time with a microbubble US contrast agent. A human uterine sarcoma cell line, FU-MMT-1, was used in vivo because this tumor is one of the most malignant neoplasms of the human solid tumors and it also has a poor response to any of the chemotherapeutic agents currently used, as well as to radiotherapy. In angiogenic inhibitors, TNP-470 was selected to use in an in vivo study, because this agent showed a higher inhibitory effect in tube formation assay in vitro, than that of FR118487, or thalidomide. The FU-MMT-1 xenografts in nude mice were treated using US at a low-intensity (2.0 w/cm(2), 1MHZ) for 4 min three times per week each after the subcutaneous injection of TNP-470 (30 mg/kg), an angiogenesis inhibitor, and this treatment was continued for 8 weeks. Either treatment of US alone or TNP-470 alone showed a suppression of tumor growth, in comparison to the non-treatment group (control), and a significantly enhanced effect was obtained using the combined treatment. A reduction in the intratumoral vascularity, which was evaluated using both color Doppler and immunohistochemistry, was significantly demonstrated using the combined treatment, in comparison to each treatment alone, and the control. No side-effect was observed in any mice in the combined treatment group. These results suggest that the antitumor effect of TNP-470 for uterine sarcoma was accelerated by US irradiation in vivo and this combination might be a potentially effective for new cancer therapy.

Topics: Angiogenesis Inhibitors; Animals; Cyclohexanes; Female; Humans; Mice; Mice, Nude; Microbubbles; O-(Chloroacetylcarbamoyl)fumagillol; Sarcoma; Sesquiterpenes; Ultrasonic Therapy; Ultrasonics; Uterine Neoplasms; Xenograft Model Antitumor Assays

We assessed the usefulness of Jun N-terminal kinase inhibitor (JNK-I) as an anti-angiogenic agent against a human uterine carcinosarcoma cell line (FU-MMT-1). JNK-I blocked FU-MMT-1-induced human arterial endothelial cell (HAEC) tube formation in an in vitro co-culture model. Cell proliferation of FU-MMT-1 or HAEC was inhibited by JNK-I. In addition, JNK-I blocked matrix metalloproteinase production but not vascular endothelial growth factor (VEGF) secretion in HAECs. Although low concentrations of JNK-I or TNP-470, an anti-cancer agent, did not separately block FU-MMT-1-induced tube formation, such tube formation was blocked by the combination of low concentrations of JNK-I and TNP-470 because TNP-470 blocked VEGF production, suggesting that JNK-I and TNP-470 had a synergistic effect and might be effective in patients with carcinosarcoma.

Topics: Angiogenesis Inhibitors; Carcinosarcoma; Cell Proliferation; Cyclohexanes; Drug Synergism; Female; Humans; JNK Mitogen-Activated Protein Kinases; MAP Kinase Kinase 4; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Mitogen-Activated Protein Kinase Kinases; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Tumor Cells, Cultured; Uterine Neoplasms; Vascular Endothelial Growth Factor A

Carcinosarcoma is the most aggressive neoplasm of among the known uterine malignancies. Most tumors show lymph-vascular space invasion and are clinically resistant to any chemotherapeutic drug currently used as well as any radiotherapy. This is the first study to investigate a novel therapeutic approach using an angiogenesis inhibitor TNP-470, a synthetic analogue of fumagillin, for human uterine carcinosarcoma in vivo.. The growth-inhibitory and anti-angiogenic effects of TNP-470 were examined after inoculating a human uterine carcinosarcoma cell line, FU-MMT-1, in nude mice. Intratumoral blood flow was evaluated weekly by color Doppler ultrasound (CDU) after the xenografts measurably developed during the period of treatment. The microvessel density (MVD) in TNP-470-treated xenografts was also immunohistochemically examined.. TNP-470 significantly reduced the volume as well as the weight of the xenografts when this therapy was started 3 weeks (Day 21) after the inoculation of FU-MMT-1, in comparison to the controls. Neither weight loss nor ataxia was observed in any mice of this therapy. A main feeding artery for the xenograft was detected by CDU in all mice treated in this study. However, no significant difference in either the vessel density visualized by CDU or MVD between the TNP-470-treated xenografts and controls was observed.. These results suggest that TNP-470 may inhibit the growth of human uterine carcinosarcoma in vivo. We speculate that TNP-470 may be a useful agent for adjuvant therapy in patients with advanced or recurrent uterine carcinosarcomas. However, a further evaluation in molecular level of the anti-angiogenic effect of TNP-470 against this tumor in vivo is thus called for.

Topics: Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Carcinosarcoma; Cell Division; Cyclohexanes; Female; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Tumor Cells, Cultured; Uterine Neoplasms; Xenograft Model Antitumor Assays

To investigate the effects of angiogenesis inhibitor TNP-470 on uterine microvessels in mice. Pituitary grafting frequently induced uterine adenomyosis.. In vivo experimental study.. Department of Biological Sciences, University of Tokyo and Medical Research Institute, Tokyo Medical and Dental University.. SHN mice, which are known to develop uterine adenomyosis spontaneously, and also very soon after pituitary grafting.. Immunohistochemical study on uterine blood vessels using an antibody to von Willebrand factor in pituitary gland-implanted mice with or without TNP-470.. Reduced incidence of uterine adenomyosis.. Twelve of 15 mice developed uterine adenomyosis with dilated blood vessels, but none of the TNP-470-treated mice with shrunken microvessels. The number of bromodeoxyuridine immunoreactive cells and activities of thymidylate synthase and thymidine kinase in uterine tissues were markedly reduced in TNP-470-treated mice.. TNP-470, a potent inhibitor of the development of vascular endothelium, reduced the development of endometrial blood vessels resulting in a lowered incidence of uterine adenomyosis induced by pituitary grafting in mice, and reduced the increase in S-phase cells and enzyme activity for pyrimidine nucleotide synthesis.

Topics: Adenomyoma; Angiogenesis Inhibitors; Animals; Body Weight; Bromodeoxyuridine; Cyclohexanes; Estrous Cycle; Female; Immunohistochemistry; Mice; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Organ Size; Random Allocation; Sesquiterpenes; Thymidine Kinase; Thymidylate Synthase; Uterine Neoplasms