o-(chloroacetylcarbamoyl)fumagillol and Liver-Neoplasms

Hepatocellular carcinoma (HCC) is a typical hypervascular tumor. Many angiogenic factors have been studied in HCC, and several anti-angiogenic therapies have been tested in animal models and patients. This paper summarizes the latest findings, especially regarding the clinical significance of endothelial cell markers and angiogenic factors in HCC, and experimental and clinical anti-angiogenesis therapies. Further developments in this area, such as endothelial cell-oriented research and better experimental and clinical designs in the evaluation of anti-angiogenic therapies are discussed.

Topics: Angiogenesis Inducing Agents; Angiogenesis Inhibitors; Angiopoietins; Animals; Antineoplastic Agents; Carcinoma, Hepatocellular; Chemotherapy, Adjuvant; Clinical Trials as Topic; Cyclohexanes; Endostatins; Fibroblast Growth Factor 2; Humans; Interferons; Interleukin-8; Liver Neoplasms; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Radiotherapy, Adjuvant; Research Design; Sesquiterpenes; Somatostatin; Vascular Endothelial Growth Factor A

The fact that tumor growth and metastatic spread relies on angiogenesis has been widely proven and accepted. The understanding of cancer biology and metastasis formation has led to the development of new therapeutic approaches that target tumor biology. The survival and establishment of metastatic lesions depend on a shift in the normal balance of proangiogenic and antiangiogenic factors that favor angiogenesis. Colorectal cancer is one of the leading cancer deaths worldwide. Angiogenesis has been associated with colon cancer progression and metastatic spread, thereby significantly affecting patient survival. New experimental approaches that inhibit angiogenic processes have demonstrated promising antineoplastic effects on metastatic colorectal cancer and are partially being investigated in clinical trials. This review focuses on angiogenesis in colorectal cancer metastasis formation as a target for antiangiogenic therapy, describing the experience from experimental studies and current clinical trials.

Topics: Angiogenesis Inhibitors; Animals; Colonic Neoplasms; Cyclohexanes; Endothelial Growth Factors; Humans; Liver Neoplasms; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Spiro Compounds; Thrombospondins; Vascular Endothelial Growth Factor A; Wound Healing

Hepatocellular carcinoma (HCC) is one of the most frequent malignancies worldwide. A variety of pharmacological strategies has been evaluated in the treatment of HCC: classical chemotherapy, tamoxifen, octreotide, thymostimulin, pravastatin, (131)I-lipiodol as well as transarterial chemoperfusion (TAC) and chemoembolisation (TACE). TACE monotherapy or TACE combined with pravastatin resulted in a survival benefit of selected HCC patients. New strategies such as immunotherapy, antiangiogenic agents or cyclooxygenase inhibitors are under clinical investigation and might play a role in future therapies for HCC. Efficient strategies for the primary prevention of HCC are available and promising concepts in the secondary prevention have been reported.

Topics: Adjuvants, Immunologic; Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Anticholesteremic Agents; Antineoplastic Agents, Hormonal; Carcinoma, Hepatocellular; Chemoembolization, Therapeutic; Cyclohexanes; Disease Models, Animal; Genetic Therapy; Humans; Immunotherapy; Liver Neoplasms; Meta-Analysis as Topic; Mice; Multicenter Studies as Topic; O-(Chloroacetylcarbamoyl)fumagillol; Octreotide; Pilot Projects; Pravastatin; Primary Prevention; Prognosis; Randomized Controlled Trials as Topic; Risk Factors; Sesquiterpenes; Tamoxifen; Thymus Extracts; Time Factors

Topics: Angiogenesis Inhibitors; Animals; Carboplatin; Carcinoma, Hepatocellular; Chemotherapy, Adjuvant; Cyclohexanes; Disease Models, Animal; Drug Therapy, Combination; Humans; Liver Neoplasms; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes

Angiogenesis is essential for the growth of solid tumors. Antiangiogenic therapy has therefore attracted considerable interest as a novel therapy for various tumors including colorectal carcinoma. We experimentally investigated the therapeutic effect of TNP-470, a nonspecific inhibitor of angiogenic factors, and vascular endothelial growth factor (VEGF)-neutralizing antibody (VEGFAb), was a VEGF-specific inhibitor, on liver metastases of colon carcinoma using a murine orthotopic transplantation model. TK-4 and TK-13 are moderately differentiated adenocarcinoma strains established in our department which express VEGF mRNA and protein. Administration of TNP-470 30 mg/kg significantly inhibited the liver metastases of both strains, as did administration of VEGFAb 100 micrograms/mouse. The therapeutic effect on liver metastases was more dominant with antiangiogenic therapy than with chemotherapy (mitomycin C). Furthermore, the sustained effect of TNP-470 induced tumor dormancy and consequently improved the survival of the animals. These results suggest that antiangiogenic treatment will be a potent therapy for liver metastases of human colorectal carcinoma in the future.

Topics: Animals; Antibiotics, Antineoplastic; Antibodies; Colorectal Neoplasms; Cyclohexanes; Disease Models, Animal; Endothelial Growth Factors; Humans; Liver Neoplasms; Lymphokines; Mice; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

The objective of this study was to determine in a rat model of hepatocarcinoma (HCC) the effects of the antiangiogenic agent TNP-470 on antioxidant enzymes, including catalase (CAT), superoxide dismutases (Mn-SOD and Cu,Zn-SOD), and glutathione peroxidase (GPx). Tumor was induced in male Wistar rats by diethylnitrosamine and promoted by two-thirds hepatectomy plus acetaminofluorene administration. Experiments were carried out 28 weeks after initiating the treatment. TNP-470 was administered at 30 mg/kg, 2 times per week from weeks 20 to 28. Carcinomatous tissue was growing outside dysplastic nodules in rats with HCC. HCC caused oxidative stress demonstrated by increased lipid peroxidation and oxidized/reduced glutathione ratio that was accompanied by a reduced activity of antioxidant enzymes Cu,Zn-SOD, GPx, and CAT. In contrast, Mn-SOD activity and expression were higher in hepatocarcinoma than in control groups. These effects were absent in animals receiving TNP-470. No significant differences between untreated and TNP-470-treated rats were observed in the expression of the Cu,Zn-SOD, glutathione peroxidise, and CAT. We conclude that TNP-470 inhibits expression and activity of Mn-SOD induced by experimental hepatocarcinogenesis. Oxidative stress reduction by TNP-470 accounts for yet another anti-cancer effect of this molecule.

Topics: Angiogenesis Inhibitors; Animals; Antioxidants; Carcinogens; Carcinoma, Hepatocellular; Cyclohexanes; Diethylnitrosamine; Disease Models, Animal; Free Radical Scavengers; Liver; Liver Neoplasms; Male; O-(Chloroacetylcarbamoyl)fumagillol; Oxidative Stress; Rats; Rats, Wistar; Sesquiterpenes

Inhibition of angiogenesis is emerging as a promising strategy for the treatment of cancer. In our study reported here, the effects of 4 highly potent methionine aminopeptidase 2 (MetAP2) inhibitors, IDR-803, IDR-804, IDR-805 and CKD-732 (designed by structure-based molecular modeling), on angiogenesis and tumor growth were assessed. Concentrations of these inhibitors as low as 2.5 nM were able to inhibit the growth of human umbilical vein endothelial cells (HUVEC) by as much as 50%, arresting growth in the G1 stage of mitosis. An intracellular accumulation of p21(WAF1/Cip1) protein was also observed. Furthermore, at higher concentrations (25 nM) of these 4 MetAP2 inhibitors, a significant induction of apoptosis was apparent in the same HUVEC cultures. As a result of these findings, the possible anticancer effects of these inhibitors were examined, utilizing the SNU-398 hepatoma cell line. Interestingly, pretreatment with these inhibitors led to an increased number of apoptotic cells of up to 60% or more, compared to untreated controls. Moreover, utilizing an in vivo xenografted murine model, these inhibitors suppressed the growth of engrafted tumor. In conclusion, these 4 inhibitory compounds potently exert an antiangiogenic effect to inhibit the growth of cancers in vivo and could potentially be useful for the treatment of a variety of cancers.

Topics: Aminopeptidases; Angiogenesis Inhibitors; Animals; Apoptosis; Blotting, Western; Carcinoma, Hepatocellular; Cell Line, Tumor; Cinnamates; Cyclohexanes; Disease Models, Animal; Endothelial Cells; Epoxy Compounds; Fatty Acids, Unsaturated; Humans; Liver Neoplasms; Metalloendopeptidases; Mice; Mice, Nude; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Transplantation, Heterologous; Umbilical Veins

Recently, 14-member macrolide antibiotics such as clarithromycin and roxithromycin have been shown to have anticancer and antiangiogenic effects. We investigated the suppressive effect of roxithromycin on accelerated hepatocellular carcinoma growth in a rat hepatocarcinogenetic model and compared results with effects from TNP-470.. Tumor was induced by oral diethylnitrosamine administration for 17 weeks. Normal saline, TNP-470 (50 mg/kg), or roxithromycin (40 or 100 mg/kg) was administered i.p. thrice per week from week 10 to 17.. Carcinomatous tissue growing outside dysplastic nodules and a marked expression of placental glutathione S-transferase were detected in rats with induced carcinogenesis. Tumor growth was accompanied by augmented expression of inducible nitric oxide synthase, activation of nuclear factor kappaB, and increased lipid peroxidation level. All these effects were absent in animals that received roxithromycin or TNP-470. The inhibitory effect of roxithromycin was dose dependent and no clear differences were noted between groups given roxithromycin 100 mg/kg and TNP-470 50 mg/kg.. Our results indicate that roxithromycin inhibits oxidative stress, nitric oxide production, and nuclear factor kappaB activation induced by experimental hepatocarcinogenesis. The data provide additional evidence for the potential use of roxithromycin in treatment of hepatocellular carcinoma prevention.

Topics: Animals; Anti-Bacterial Agents; Antineoplastic Agents; Carcinogens; Carcinoma, Hepatocellular; Cell Nucleus; Cyclohexanes; Diethylnitrosamine; Disease Models, Animal; Dose-Response Relationship, Drug; Glutathione Transferase; Lipid Peroxidation; Liver; Liver Neoplasms; Male; Models, Biological; NF-kappa B; Nitric Oxide Synthase; O-(Chloroacetylcarbamoyl)fumagillol; Oxidative Stress; Placenta; Rats; Rats, Wistar; Roxithromycin; Sesquiterpenes

We investigated the suppressive effect of the angiogenesis inhibitor TNP-470 on accelerated hepatocellular carcinoma (HCC) growth in the regenerating liver.. After 70% partial hepatectomy (PH), AH-130 cells were injected into the portal vein of Donryu rats. A control group was given the vehicle only, and the treated group was given 10 mg/kg TNP-470 subcutaneously every second day, from 24 h after tumor implantation, seven times. On day 14, tumor growth was evaluated by the number of foci on the liver surface, liver weight, and the microvessel density of the tumor.. The number of foci was significantly less in the treated group (116.5 +/- 103.1) than in the control group (319.3 +/- 223.1) ( P < 0.05), as was microvessel density, which was 31.3 +/- 14.0/mm2 in the treated group and 61.2 +/- 18.9/mm2 in the control group ( P < 0.05). The liver tended to weigh less in the treated group (12.15 +/- 1.28 g) than in the control group (15.22 +/- 5.35 g). We also assessed whether TNP-470 retards liver regeneration. Seven days after 70% PH, the liver weight in the treated group was similar to that in the control group. Total bilirubin, serum glutamic oxaloacetic transaminase, and serum glutamic pyruvic transaminase were not higher in the treated group than in the control group.. TNP-470 can suppress HCC growth without retarding liver regeneration after PH.

Topics: Angiogenesis Inhibitors; Animals; Carcinoma, Hepatocellular; Cyclohexanes; Hepatectomy; Liver Neoplasms; Liver Regeneration; Male; Neoplasm Recurrence, Local; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Sesquiterpenes; Tumor Cells, Cultured

The objective of this study is to determine if treatment with the angiogenesis inhibitor TNP-470 results in impairment of oxidative stress, inhibition of nuclear factor kappa B (NF-kappaB) activation and decrease of nitric oxide production in an experimental model of rat hepatocarcinogenesis. Tumour was induced by diethylnitrosamine and promoted by two-thirds hepatectomy plus acetaminofluorene administration. Experiments were carried out at 28 weeks after initiating the treatment. TNP-470 was administered at 30mg/kg, three times per week from 20 to 28 weeks. Carcinomatous tissue growing outside dysplastic nodules and a marked expression of placental glutathione S-transferase were detected in rats with induced carcinogenesis. Liver concentrations of thiobarbituric acid reactive substances, reduced glutathione (GSH) and glutathione disulfide (GSSG) were significantly higher than those of controls and there was a significant increase in the GSSG/GSH ratio. Tumour growth was accompanied by augmented expression of inducible nitric oxide synthase, activation of (NF-kappaB) and proteolysis of IkappaB. All these effects were absent in animals receiving TNP-470. Our results indicate that TNP-470 inhibits oxidative stress, nitric oxide production and NF-kappaB activation induced by experimental hepatocarcinogenesis. These changes would contribute to the beneficial effects of TNP-470 in cancer treatment.

Topics: Animals; Antibiotics, Antineoplastic; Blotting, Western; Body Weight; Carcinoma, Hepatocellular; Cyclohexanes; Diethylnitrosamine; Enzyme Activation; Glutathione; Glutathione Disulfide; Glutathione Transferase; Humans; I-kappa B Proteins; Liver; Liver Neoplasms; Male; NF-kappa B; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; O-(Chloroacetylcarbamoyl)fumagillol; Oxidative Stress; Rats; Rats, Wistar; Sesquiterpenes; Thiobarbituric Acid Reactive Substances; Time Factors; von Willebrand Factor

An angiogenesis inhibitor, TNP-470 (TNP), has shown promising results in tumor dormancy therapy, and we have been studying its antitumor effects using a rabbit spontaneous liver metastasis model. However, because inhibition was observed only at the step of micrometastasis, we examined the effects of combining TNP in the same model with a nonspecific immunopotentiator, lentinan (LNT), as a biological response modifier.. The model was established by the inoculation of VX-2 tumors into the colon, and colectomy was subsequently performed, including the primary tumor. Combination (TNP + LNT) effects were evaluated in terms of the number and volume of metastatic nodules, microvessel density (MVD), expression of proliferating cell nuclear antigen (PCNA), and apoptosis, using immunohistochemical staining with anti-CD31, anti-PCNA monoclonal antibody, and the TUNEL (in situ nick end-labeling) method, respectively.. Angiogenesis was significantly inhibited in the TNP + LNT group, and the apoptotic index was also significantly higher than in the TNP or LNT groups. The positive expression of PCNA in the VX2 cells was reduced in the LNT alone and TNP + LNT groups, but not in the TNP alone group.. These findings indicate that TNP-470 and lentinan could prove useful for preventing the development of metachronous liver metastases from colorectal cancers after curative resection.

Topics: Angiogenesis Inhibitors; Animals; Apoptosis; Colonic Neoplasms; Cyclohexanes; Female; Immunohistochemistry; Infusions, Intravenous; Liver Neoplasms; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rabbits; Sesquiterpenes; Statistics, Nonparametric

TAC-101 (4-[3,5-bis(trimethylsilyl)benzamido]benzoic acid) is a novel, synthetic retinoid that is effective against liver metastases of human gastrointestinal cancer cells such as the human stomach carcinoma line AZ-521 in animal models, and is currently in use in phase I cancer trials. However, the mechanism of its antimetastatic action is still poorly understood. Tumor metastasis depends on angiogenesis, and various retinoids have been found to exhibit antiangiogenic activity. Based on these findings we here examined the antiangiogenic effects of TAC-101. Oral administration of TAC-101 (2-8 mg/kg/day) resulted in a drastic suppression of the AZ-521 cell-induced angiogenesis in a mouse dorsal air sac assay system, compared to the vehicle alone. Immunohistochemical analysis with antibody against the endothelial marker CD31 revealed a significant reduction in microvessel density in liver metastases from animals treated with TAC-101 (8 mg/kg p.o.), compared to liver metastases from the untreated control animals. The ability of TAC-101 (8 mg/kg p.o.) to prevent experimental liver metastasis of AZ-521 cells in athymic nude mice was comparable with that of the known angiogenesis inhibitor TNP-470 (30 mg/kg s.c.). TAC-101 also affected angiogenesis in chorioallantoic membranes and some functions of endothelial cells associated with angiogenesis, whereas the retinoid failed to suppress AZ-521 cell proliferation directly. These data suggest that the TAC-101 is an orally active antiangiogenic agent and that this antiangiogenic property may contribute to its efficacy against liver metastasis of human stomach cancer cells.

Topics: Administration, Oral; Animals; Benzoates; Cell Division; Cell Movement; Cisplatin; Cyclohexanes; Dose-Response Relationship, Drug; Endothelium, Vascular; Humans; Immunohistochemistry; Liver Neoplasms; Mice; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Platelet Endothelial Cell Adhesion Molecule-1; Sesquiterpenes; Stomach Neoplasms; Trimethylsilyl Compounds; Tumor Cells, Cultured

To prevent tumor metastasis, we administered the cell-binding domain of fibronectin, in combination with the angiogenesis inhibitor TNP-470, to mice with hepatic metastasis. We then assessed the prevention of tumor metastasis resulting from the inhibition of adhesive interactions and the inhibition of angiogenesis.. A hepatic metastasis model was created by injecting 1 x 10(3) colon 26/TC-1 cells into the anterior mesenteric vein of CDF1 mice. The cell-binding domain obtained from fibronectin included the Arg-Gly-Asp (RGD) sequence. A fibronectin-binding domain (FND)-treated group, an FND plus TNP-470 group, and a control group were established. The animals were killed 4 weeks after the injections of the treatment agents had been completed and the number of metastatic liver nodules was counted. In a simultaneous experiment with the same design, the mice were not killed at 4 weeks, and their survival was observed.. The mean number of nodules in the FND plus TNP-470 group was significantly lower than that in the control group (P = 0.019337). The inhibition rate was 51% in the FND group, 60% in the FND 10 micrograms plus TNP-470 10 mg/kg group, and 64% in the FND 10 micrograms plus TNP-470 100 mg/kg group compared with the control group. Mice from the FND group that were not killed died after 6-8 weeks, but mice from the FND plus TNP-470 group died after 8-12 weeks.. The cell-binding domain of fibronectin may, potentially, be an effective form of antiadhesive therapy that competes with native adhesion molecules and blocks adhesion during the metastatic process. When the cell-binding domain of fibronectin is combined with TNP-470 to inhibit angiogenesis, more effective inhibition of metastatic tumor growth and prolongation of survival can be achieved than after treatment with the cell-binding domain alone.

Topics: Angiogenesis Inhibitors; Animals; Colorectal Neoplasms; Cyclohexanes; Disease Models, Animal; Fibronectins; Liver Neoplasms; Male; Mice; O-(Chloroacetylcarbamoyl)fumagillol; Oligopeptides; Sesquiterpenes; Tumor Cells, Cultured

This study was undertaken in order to assess the efficacy of a potent angiogenesis inhibitor, TNP-470, on tumor growth in a syngeneic rodent model of liver metastases from colorectal cancer.. New blood vessel formation is a prerequisite for primary and metastatic tumor growth. TNP-470, a synthetic derivative of fumagillin when subcutaneously transplanted into nude mice, inhibits endothelial cell proliferation and migration, as well as the growth of various human cancers. However, the antitumor effect of this drug has not been studied in models reproducing a natural metastatic environment. Since the liver provides an extensive vascular bed for secondary tumor growth, an anti-angiogenic strategy may therefore be less efficient for treating hepatic metastases than primary tumors.. 10(7) DHD K12 colon carcinoma cells were injected intrasplenically into syngeneic BD IX rats to produce diffuse liver metastases. TNP-470 (30 mg/kg/day) was administered on alternate days starting 4 days after tumor implantation. The animals were sacrificed after 4 weeks and their livers were processed for histologic examination. In both the treatment and control groups (n=7), tumor volume was determined using a computerized analytical system, and tumor microvessel density was measured by immunostaining with anti-von Willebrand Factor monoclonal antibody.. In vitro, TNP-470 demonstrated a direct toxicity towards the DHD K12 cell line with an IC50 of 0.1 microg/ml. Metastases were present in all animals from both groups. Liver weight (15.2 g vs 11.7 g, p=0.01), and tumor volume (1218 mm3 vs 406 mm3, p=0.03) were significantly reduced in the TNP-470 group compared to the control group. Tumor microvessel density was not statistically different between the two groups (67 vs 63 microvessels/x200 field, p=0.41).. TNP-470 inhibits the growth of liver metastases in a syngeneic rat model of colorectal cancer. The mechanism responsible for this effect remains unclear, but may involve a combination of anti-angiogenic and direct cytotoxic effects.

Topics: Angiogenesis Inhibitors; Animals; Carcinoma; Cell Division; Cell Survival; Colorectal Neoplasms; Cyclohexanes; Injections, Subcutaneous; Liver; Liver Neoplasms; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Organ Size; Rats; Sesquiterpenes; Tumor Cells, Cultured

The contribution of angiogenesis to tumor growth and hepatic metastasis of colorectal cancer was investigated by means of immunohistochemical study and in vitro and in vivo experiments. Colorectal cancer specimens from 30 patients with hepatic metastasis and 39 patients without hepatic metastasis were studied by staining with antibodies against factor VIII-related antigen. Microvessel count in patients with liver metastasis was significantly higher than in those without liver metastasis (p<0.005). The effect of TNP-470 was evaluated with in vitro and in vivo experiments using human colon cancer cell line, LM and the highly hepatic metastasis cell line, LM-H5. The effect of TNP-470 on the proliferation of the cancer cells and human umbilical vein endothelial cells (HUVECs) was examined. TNP-470 inhibited more sensitively the proliferation of HUVECs than cancer cells in vitro. IC50 was approximately 3 pg/ml in HUVECs and approximately 2 microg/ml in cancer cells. The effect of TNP-470 on the growth of xenografts and liver metastases by LM-H5 in nude mice was examined. TNP-470 (30 mg/kg) was administered by subcutaneous injection every third day for 4 weeks. TNP-470 inhibited both the growth of xenograft and the hepatic metastasis. The number of metastatic foci in the liver was 78.2+/-30.1 in the control group and 20.6+/-16.5 in the treated group. These results suggest that TNP-470 is a potent agent to inhibit tumor growth and hepatic metastasis of colon cancer.

Topics: Adult; Aged; Aged, 80 and over; Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Colorectal Neoplasms; Cyclohexanes; Female; Humans; Liver Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Middle Aged; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Tumor Cells, Cultured

The effect of TNP-470, an angiogenesis inhibitor, on the growth of a hepatoblastoma transplanted into nude mice was examined.. A hepatoblastoma obtained from a 3-year-old girl was serially transplanted into nude mice subcutaneously, and the transplant tumors of the seventh and eighth generations were used for experiments. Expression of various markers in the tumors was examined immunohistochemically. TNP-470 was injected subcutaneously every other day into tumor-bearing mice from 3 weeks after tumor transplantation. The proliferation of tumor cells and endothelial cells was estimated by means of the bromodeoxyuridine labeling index.. The original hepatoblastoma showed the histology of the epithelial type, consisting of both the fetal and embryonal subtypes and was positively stained with anti-alpha-fetoprotein (AFP), anti-cytokeratin-19 and polyclonal anticarcinoembryonic antigen antibodies, and an antihuman hepatocyte antibody (hepatocyte paraffin 1). The transplant tumors consisted of solid nests of tumor cells with numerous vascular lakes of various sizes, and showed positive staining with all antibodies that reacted positively with the original hepatoblastoma. Injections of TNP-470 at the doses of 15 mg and 30 mg/kg body weight suppressed the tumor growth and the increase in the serum level of AFP dose dependently. Injections of TNP-470 also suppressed the proliferation of tumor cells and endothelial cells in the tumors.. Hepatoblastomas maintained in nude mice retained the immunohistochemical characteristics of the original hepatoblastoma, and TNP-470 suppressed the growth of hepatoblastomas transplanted into nude mice. TNP-470 may be worth investigating further as to its usefulness as a therapy for hepatoblastomas.

Topics: alpha-Fetoproteins; Angiogenesis Inhibitors; Animals; Child, Preschool; Cyclohexanes; Female; Hepatoblastoma; Humans; Immunohistochemistry; Liver Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Transplantation, Heterologous

Even though angiogenesis inhibitor is thought to be one of the promising agents in tumor dormancy therapy, its optimal administration is still unknown. Therefore, the efficient protocol using TNP-470 was examined regarding its treatment affect against spontaneous liver metastases of colon tumors in the rabbit. A spontaneous liver metastases model was established in the rabbit by the inoculation of VX-2 tumors into the subserosal space of the colon. The therapeutic effect of TNP-470 was then investigated by monitoring both the number of metastatic nodules in the liver and the microvessel density (MVD) in the tumor by immunohistochemical staining using anti-CD31 monoclonal antibody. TNP-470 did not show any effect on the primary tumor. It was able to reduce the metastatic spread to liver when it was administered at the microscopic metastatic stage. Treatment at this stage, however, was not able to sufficiently control the disease. These results indicated that TNP-470 could efficiently cause the tumor to enter into a dormant state by inhibiting angiogenesis when it was used at the initial stage of the metastatic process in the liver. Regarding its clinical application, TNP-470 might be useful for preventing the metachronous liver metastases of colorectal cancer when it is administered as adjuvant therapy after curative surgery.

Topics: Angiogenesis Inhibitors; Animals; Colonic Neoplasms; Cyclohexanes; Female; Infusions, Intravenous; Liver Neoplasms; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rabbits; Sesquiterpenes

The antitumor and antimetastatic effects of TNP-470, an angiogenesis inhibitor, on human gastrointestinal tumors xenotransplanted into nude mice were investigated. When two gastric cancer (MT-2 and MT-5) and two colon cancer (TK-4 and TK-13) xenografts are transplanted orthotopically into nude mice, liver metastasis develops 6 weeks after transplantation. TNP-470 30 mg/kg had a significant inhibitory effect on primary tumor growth of gastric cancers when given on alternate days from 7 days after transplantation. However, when given from 10 days or 14 days after transplantation, no inhibitory effect on the growth of any tumor xenograft was observed. In contrast, liver metastasis of each xenograft type was inhibited significantly by TNP-470. The effect of TNP-470 on prognosis was investigated using a hepatic metastatic model of rat hepatoma. Although all untreated rats that received AH-130 cell implants died within one month of massive hepatic metastasis, >50% of rats treated with TNP-470 survived for 4 months. The number of apoptotic cells in hepatic metastatic foci was significantly increased by TNP-470 administration. These results suggest that TNP-470 may provide a new approach to the treatment of digestive organ malignancies.

Topics: Animals; Anti-Bacterial Agents; Antibiotics, Antineoplastic; Body Weight; Cyclohexanes; Digestive System Neoplasms; Fatty Acids, Unsaturated; Humans; Liver Neoplasms; Liver Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Organ Size; Rats; Sesquiterpenes; Transplantation, Heterologous

Combination therapy using HCFU and TNP 470, which inhibits angiogenesis, was examined for effectiveness against the footpad injection model of LMFS tumor. This LMFS, a retroperitoneal sarcoma of BALB/c mice, proliferated at the inoculation site (100% take) and all mice operated on after day 15 had spontaneous metastatic nodules in the liver. Mice with the LMFS tumor were given HCFU and 5-FU (5 days/week), and/or TNP 470 (3 days/week) from day 3 for 3 weeks and sacrified at day 28 under anesthesia. Seven of 10 mice receiving 5-FU and TNP 470 died from the side effects of the drugs. Mean tumor weight and liver metastatic nodules were determined and compared with a control group. The results were as follows: HCFU group: 94.6%, 11.8%, 5 FU group: 73.9%, 28.8%, TNP 470 group: 67.6%, 44%, HCFU and TNP 470 group: 33.3%, 6.4%. Mice with LMFS were given HCFU and/or TNP 470 from day 3 for 4 weeks. The foot on the injected side was amputated on day 15, and the animals were sacrified on day 35. Liver metastatic nodules compared with those of the operation (OP) group as follows: OP + HCFU group: 14.4%, OP + TNP group: 39.1%, and OP + HCFU + TNP 470 group: 5.4%. Histologically, 5 of 5 mice of the OP group, 3 of 5 of the OP + HCFU group, 4 of 5 of the OP + TNP 470 group and 1 of 5 of the OP + HCFU + TNP 470 group had liver metastases. These results show that while either HCFU or TNP 470 is effective by itself, a combination of these drugs is more effective against liver metastasis.

Topics: Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Antineoplastic Agents; Cyclohexanes; Female; Fluorouracil; Liver Neoplasms; Mice; Mice, Inbred BALB C; O-(Chloroacetylcarbamoyl)fumagillol; Retroperitoneal Neoplasms; Sarcoma, Experimental; Sesquiterpenes; Tumor Cells, Cultured

TNP470, a derivative of fumagillin, suppressed in vivo growth of human PLC/PRF/5 hepatoma and ameliorated cachexia of hepatoma-bearing mice. These in vivo effects were associated with reductions in microvessel and macrophage counts. In in vitro experiments, TNP470 inhibited the growth and migration of human hepatoma and bovine vascular endothelial (VE) cells. TNP470 did not inhibit the production of VE growth factor by the hepatoma, which suggests that this compound acts directly on VE cells in vivo. In contrast, TNP470 inhibited the production of leukemia inhibitory factor, which may be related to the amelioration of cancer cachexia. TNP470 induced apoptosis and enhanced the expression of beta-galactosidase, a biomarker of senescence, which was partly mimicked by a nitric oxide (NO) donor S-nitroso-N-acetyl penicillamin. TNP470 inhibited myristoylation and membrane translocation of NO synthase and increased the cellular content of NO synthase and production of NO. Therefore, it is suggested that the actions of TNP470 are mediated, at least in part, through the inhibition of membrane translocation of biologically active proteins.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; beta-Galactosidase; Carcinoma, Hepatocellular; Cattle; Cell Division; Cell Movement; Cyclohexanes; Endothelial Growth Factors; Endothelium, Vascular; Humans; Liver Neoplasms; Lymphokines; Mice; Mice, Nude; Neovascularization, Pathologic; Nitric Oxide; Nitric Oxide Synthase; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Transplantation, Heterologous; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors; Weight Loss

The anti-tumor and anti-metastatic effects of O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), an angiogenesis inhibitor, and cisplatin (CDDP), an anti-neoplastic agent, were investigated using our established liver-metastasizing pancreatic carcinoma line, HPC-3H4. HPC-3H4 was injected into the spleens of nude mice. Mice were randomly divided into 5 groups; a control group given saline solution, a group receiving 45 mg/kg TNP-470, a group receiving 90 mg/kg TNP-470, a group receiving 90 mg/kg TNP-470 in combination with 0.25 mg/kg CDDP, and a group receiving 0.25 mg/kg CDDP. In the control group, liver metastasis developed in 14 of 15 mice (93.3%). Liver metastasis developed in 9 of 11 mice (81.8%) receiving 0.25 mg/kg CDDP. It developed in 11 of 15 mice (73.3%) receiving 45 mg/kg TNP-470, in 17 of 18 mice (94.4%) receiving 90 mg/kg TNP-470, and in 4 of 10 mice (40%) receiving 90 mg/kg TNP-470 in combination with 0.25 mg/kg CDDP. TNP-470 in combination with CDDP displayed a significant inhibitory effect on liver metastasis compared to the control. Although TNP-470 alone and CDDP alone had no effect on the tumor growth in vivo, 90 mg/kg TNP-470 in combination with 0.25 mg/kg CDDP had a significant effect. In vitro examinations demonstrated that the growth of HPC-3H4 cells was only mildly inhibited by TNP-470, but the production of vascular endothelial growth factor (VEGF) by HPC-3H4 was clearly inhibited by TNP-470. The inhibitory effect on the production of VEGF was not strong with CDDP treatment. These results indicate that the angiogenesis inhibitor TNP-470 in combination with low-dose CDDP has inhibitory activity against liver metastasis of human pancreatic carcinoma.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Cell Division; Cisplatin; Cyclohexanes; Endothelial Growth Factors; Female; Humans; Liver Neoplasms; Lymphokines; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Pancreatic Neoplasms; Sesquiterpenes; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

To study the effect of angiogenesis inhibitor TNP-470 on the growth and metastasis of gastric cancer in vivo.. Metastatic model simulating human gastric cancer was established by orthotopic implantation of histologically intact human tumor tissue into gastric wall of nude mice. TNP-470 was administered s.c. at doses of 0 mg/kg, 15 mg/kg, 30 mg/kg, 60 mg/kg every other day for eight weeks. Ten weeks after implantation, the mice were sacrificed and the tumor size measured and the presence of metastasis recorded. The microvascular density was examined by immunohistochemical staining with anti-human factor VIII antibody.. Compared to the untreated controls, growth of the orthotopically implanted tumor was significantly reduced in size in mice treated with TNP-470 with an inhibition rate of 59.9%, 77. 0% and 84.9% at the dosage of 15 mg/kg, 30 mg/kg and 60 mg/kg, respectively. Tumor metastasis to the liver and peritoneaum was also significantly inhibited in a dose-dependent manner. The microvascular density was also decreased significantly in the treated mice.. Angiogenesis inhibitor TNP-470 has strong inhibitory effect both on tumor growth and metastasis of human gastric cancer in nude mice.

Topics: Animals; Antibiotics, Antineoplastic; Cyclohexanes; Female; Humans; Liver Neoplasms; Mice; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Peritoneal Neoplasms; Sesquiterpenes; Stomach Neoplasms; Tumor Cells, Cultured

The effect of an angiogenesis inhibitor, TNP-470, on primary tumor growth, liver metastasis and peritoneal dissemination of gastric cancer was investigated by means of an orthotopic xenotransplanted model of 2 human gastric cancers, MT-2 and MT-5. TNP-470 showed a significant inhibitory effect on the growth of primary tumors after orthotopic transplantation of both xenografts when given at a dose of 30 mg/kg on alternate days from day 7 after transplantation (early treatment). However, growth of the MT-2 primary tumor was not inhibited by administration from day 14 after transplantation (late treatment). Liver metastasis was prevented significantly by early treatment of TNP-470. In particular, early treatment of MT-2 completely inhibited the development of macroscopic foci in the liver and was significantly more effective than late treatment. Peritoneal dissemination also was inhibited. Thus, TNP-470 was revealed to have strong inhibitory activity not only on primary tumors and liver metastases but also against peritoneal dissemination. These results suggest that this agent may provide a new approach to the treatment of gastric cancer.

Topics: Adenocarcinoma; Animals; Antibiotics, Antineoplastic; Body Weight; Cell Division; Cyclohexanes; Humans; Liver Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Peritoneal Neoplasms; Sesquiterpenes; Stomach Neoplasms; Transplantation, Heterologous; Tumor Cells, Cultured

The antitumor and anti-metastatic effects of a potent angiogenesis inhibitor, O-(chloroacetyl-carbamoyl)fumagillol (TNP-470), was investigated in a highly metastatic model of human hepatocellular carcinoma-LCI-D20. Small pieces of LCI-D20 tumor tissue were implanted subcutaneously into the right axillary region of 24 nude mice; the mice were then randomized into two groups. To one group, TNP-470 30 mg/kg was given as a subcutaneous injection every other day from day 1 to day 15 and the mice were sacrificed on day 26. An antitumor effect of TNP-470 was clearly demonstrated by tumor weight (0.97 +/- 0.34 g compared to 2.04 +/- 0.34 g, P < 0.001) and alpha-Fetoprotein value (93 +/- 59 micrograms/L compared to 769 +/- 282 micrograms/L, P < 0.001). There was also an anti-metastatic effect of TNP-470. Lung metastases developed in only 1 of 12 mice in the treated group, while they developed in 6 of mice of the control group. No severe side-effect of TNP-470 was found in this study. In vitro study revealed that the purified hepatoma cells were insensitive to TNP-470 (the 50% inhibitory concentration was 43 micrograms/ml). These results suggest that the angiogenesis inhibitor TNP-470 has both strong antitumor and anti-metastatic effects on a human hepatocellular carcinoma model in nude mice.

Topics: Animals; Antibiotics, Antineoplastic; Cyclohexanes; Humans; Liver Neoplasms; Liver Neoplasms, Experimental; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Metastasis; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes

We examined the mechanism of the reduction of metastatic liver tumors in rats treated with angiogenesis inhibitor TNP-470 on the histological basis of patterns of tumor cell death. Metastatic tumors were developed by intravenous injection of AH-130 cell line, followed by a dose of TNP-470. Alteration in the size and number of metastatic liver tumors and its cell death pattern were analyzed. The number and size of the metastatic tumors at 2 weeks in untreated rats were larger than those in treated rats. The number of tumors in untreated rats decreased, whereas their size increased. All rats treated were alive and free from tumors after 4 weeks, although all untreated rats died of metastatic tumors. Necrosis was predominant in the tumors of untreated rats, while most tumors in treated rats showed apoptosis. Consequently, the metastatic tumor in untreated rats might grow faster than its angiogenesis, suggesting the occurrence of central necrosis due to apparent ischemia. On the other hand, the tumor in treated rats might be reduce a by weak ischemic stimulus, which triggers apoptosis.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; Cell Division; Cyclohexanes; Liver Neoplasms; Necrosis; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Sesquiterpenes

Effects on the liver of the antiangiogenesis agent O-(chloroacetyl-carbamoyl) fumagillol (TNP-470) on hematogenous metastasis of a human pancreatic carcinoma cell line were examined. One million PCI-43 cells, a human pancreas carcinoma cell line, were injected into the spleen of SCID beige mice, then TNP-470 at 30 mg/kg was administered subcutaneously every other day. The mice were killed 6 or 10 weeks thereafter and metastatic nodules in the liver were counted and measured microscopically. Metastases were inhibited and an approximately 10% loss of weight was evident in the TNP-470-administered mice. There was no suppression in maximal size of metastatic colonies in mice given the agent for 6 weeks, while inhibition was apparent in mice given the drug for 10 weeks. Suppression of proliferation and an increase in apoptosis were evident in metastatic nodules in the TNP-470-administered groups, following stainings for proliferative cell nuclear antigen and terminal deoxytransferase-mediated dUTP-biotin nick end-labeling, respectively. TNP-470 inhibited the proliferation of human umbilical vein endothelial cells but not PCI-43 in vitro. TNP-470 did not suppress production of vascular endothelial growth factor by PCI-43 cells. Neovascularization in vivo induced by PCI-43 cells was suppressed in the TNP-470-administered mice, using a diffusion chamber placed in subcutaneous tissues of SCID beige mice. These observations suggest that inhibition of angiogenesis is effective in suppressing establishment and subsequent growth of hematogenous micrometastases of pancreatic adenocarcinoma to the liver.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; Cell Division; Cyclohexanes; Endothelial Growth Factors; Humans; Liver Neoplasms; Lymphokines; Mice; Mice, SCID; Neoplasm Metastasis; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Pancreatic Neoplasms; Sesquiterpenes; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

To clarify the mechanism of the reduction of metastatic liver tumors in rats treated with angiogenesis inhibitor TNP-470, the death of tumor cells was examined pathologically and ultrastructurally. Liver metastases were developed by intravenous injection of AH-130 cells. TNP-470 was given subcutaneously after tumor cell injection. Alterations in the size and number of metastatic tumors were examined at various time points, in association with the analysis of cell death pattern. The metastatic nodules were divided into 4 groups according to the morphological patterns of cell death; no cell death, scattered apoptosis, central necrosis, and diffuse necrosis. The number and size of the metastatic tumors at 2 weeks in untreated rats were larger than those in treated rats. The number of tumors in untreated rats decreased, but the tumor size increased. All rats treated with TNP-470 were alive and free from tumors after 4 weeks, whereas all the untreated rats died of liver metastases. The percentages of the tumors with necrosis in untreated rats (61.2% at 2 weeks and 100% at 4 weeks) were significantly higher than that (31.8% at 2 weeks) in treated rats (P < 0.01). The percentage of the tumors containing apoptotic cells in treated rats was significantly higher than that in untreated rats (54.5% vs. 30.6%; P < 0.05). The growth of metastatic tumors without treatment might be faster than the growth of vessels in untreated tumors, resulting in central necrosis due to ischemia. On the other hand, the reduction of metastatic liver tumors treated with TNP-470 might be caused by inhibition of angiogenesis, providing a weak ischemic stimulus which triggers apoptosis, rather than by a direct cytotoxic effect on tumor cells, because previous in vivo experiments demonstrated that TNP-470 affected endothelial cells but not tumor cells.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; Cyclohexanes; Liver; Liver Neoplasms; Male; Microscopy, Electron; Necrosis; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Rats, Inbred Strains; Remission Induction; Sesquiterpenes; Tumor Cells, Cultured

The summation of gene mutations increases the metastatic potential of colorectal cancer. The genetic characterization and hepatic metastatic potential of five xenotransplanted human colon carcinoma strains were investigated. Furthermore, the therapeutic effect of the angiogenesis inhibitor, TNP-470, was evaluated.. The correlation between gene mutation and rate of hepatic metastases of five colon cancer strains transplanted orthotopically or subcutaneously was evaluated. The strain with the highest hepatic metastatic rate from orthotopical tumors, TK-4, was used in the experiment with TNP-470 treatment. Mice were given tumor transplants orthotopically or subcutaneously followed by 30 mg/kg of TNP-470 on alternate days from Day 10 or Day 21 after transplantation, respectively.. The rate of hepatic metastases from orthotopically transplanted tumors of 5 strains was 38 to 79%. Interestingly, TK-4 with K-ras and p53 mutations and overexpression of p53 protein induced hepatic metastases from both orthotopical (79%) and subcutaneous tumors (44%). Although TNP-470 only significantly inhibited subcutaneous tumor growth, its antimetastatic effect was significantly demonstrated on the hepatic metastases of both orthotopical and subcutaneous tumors.. p53 mutation is thought to enhance angiogenesis, favoring the growth of the hepatic metastases. TNP-470 proved the excellent antimetastatic effect of TK-4 on hepatic metastases. TK-4 has the highest metastatic potential and p53 mutation. An antiproliferative effect was observed on the rapidly growing primary tumors in which angiogenesis may be dominant.

Topics: Animals; Antibiotics, Antineoplastic; Body Weight; Cell Cycle; Cell Division; Colorectal Neoplasms; Cyclohexanes; Genes, p53; Genes, ras; Humans; Immunohistochemistry; Liver Neoplasms; Male; Mice; Mice, Inbred BALB C; Mutation; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Transplantation, Heterologous; Tumor Cells, Cultured

Effects of a newly developed angiogenesis inhibitor, TNP-470 (AGM-1470) alone and in combination with cisplatinum on tumor-bearing rats were investigated. Wistar-King A male rats were subcutaneously inoculated with 10(7) cultured syngeneic liver cancer cells that had been induced by oral intake of 3'-methyl-4-dimethylaminoazobenzen, and were used as tumor-bearing rats. Tumor sizes and changes in serum copper levels were measured after administration of TNP-470. Administration of TNP-470 (5, 10, 20 or 30mg/kg, s.c., daily for 7 days) inhibited tumor growth in a dose-dependent manner. The suppressive effects were the same in the rats which were administered TNP-470 (20mg/kg, s.c., daily for 7 days) and cisplatinum (0.5 mg/kg, s.c., daily for 7 days) simultaneously, and rats administered cisplatinum (0.5mg/kg, s.c., daily for 7 days) following TNP-470 (20mg/kg, s.c., daily for 7 days). These inhibitory effects were almost the same as that of cisplatinum (0.5mg/kg, s.c., daily for 7 days) alone. While, administration of TNP-470 (20mg/kg, s.c., daily for 7 days) following cisplatinum treatment (0.5 mg/kg, s.c., daily) showed markedly higher anti-tumor effects, compared with these groups. Administration of TNP-470 caused elevation of serum copper levels in normal rats as well as tumor-bearing rats with the same degree. Serum copper levels remained normal after discontinuation of TNP-470 in normal rats, while in tumor-bearing rats, it decreased during the first week and re-elevated in 2 to 3 weeks after discontinuation of TNP-470. This re-elevation of serum copper levels was related to rapid tumor growth after discontinuation of TNP-470. Furthermore, there was also a positive correlation between serum copper levels and capillary density in the tumor. In conclusion, TNP-470 had anti-tumor effect in a dose-dependent manner against a rat liver cancer, with markedly higher effects when it was administered following cisplatinum. Serum copper levels after discontinuation of TNP-470 treatment might indicate re-proliferation of the capillaries in the tumor tissue.

Topics: Animals; Antibiotics, Antineoplastic; Antineoplastic Combined Chemotherapy Protocols; Cell Division; Cisplatin; Copper; Cyclohexanes; Depression, Chemical; Liver Neoplasms; Male; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Rats, Wistar; Sesquiterpenes

An excellent therapeutic effect of angiogenesis inhibitors on tumor growth or metastasis has been reported, but the sustained antimetastatic effect of these agents has not been studied. We investigated the sustained effect of TNP-470, an angiogenesis inhibitor, in rats with hepatic metastasis following intraportal implantation of rat ascites hepatoma AH-130 cells. TNP-470 was administered subcutaneously at 15 mg/kg (L-TNP) or 30 mg/kg (H-TNP) on alternate days for 2 weeks. The number of liver metastases was significantly reduced in both the L-TNP (85.1 +/- 77.6) and H-TNP (31.7 +/- 49.6) groups compared to the control group (300.7 +/- 100.7) (P < 0.01) at 14 days after the start of treatment. Although all rats in the control group died within 1 month of massive liver metastasis, the L-TNP and H-TNP, respectively, had a survival rate of 82 and 60%, at 4 months (P < 0.001). Absence of toxicity of TNP-470 at the lower dose, as evidenced by the absence of intraperitoneal or intrapleural bleeding, contributed to a better prognosis in the L-TNP group. Interestingly, a small dormant metastatic focus was found in only 1 of 15 rats surviving for 4 months, whereas metastatic foci were observed in all rats at the end of treatment. These results suggest that the sustained cytostatic effect of TNP-470 on vascular endothelial cells may help to improve long-term survival by reducing the metastatic burden.

Topics: Animals; Antibiotics, Antineoplastic; Body Weight; Carcinoma, Hepatocellular; Cyclohexanes; Follow-Up Studies; Liver; Liver Neoplasms; Lymphatic Metastasis; Male; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Rats, Inbred Strains; Sesquiterpenes; Survival Analysis; Time Factors

The anti-tumor and anti-metastatic effects of TNP-470, an angiogenesis inhibitor, and mitomycin C (MMC), a representative anti-neoplastic agent, were investigated using our established liver-metastasizing gastric carcinoma line, AZ-H5c. AZ-H5c was injected into the spleen of nude mice which had been randomly divided into 4 groups; a control group given saline solution, a group receiving 15 mg/kg TNP-470, a group receiving 30 mg/kg TNP-470 and a group receiving 2 mg/kg MMC. TNP-470 was given s.c. on alternate days for 5 weeks from day 10 after intrasplenic injection, and MMC was administered intraperitoneally (i.p.) once a week from day 10 after intrasplenic injection. In the control group, liver metastasis developed in 13 of 16 mice (81%). Liver metastasis developed in 6 of 11 mice (55%) receiving MMC. In contrast, liver metastasis developed in 4 of 8 mice (50%) receiving 15 mg/kg TNP-470, and in 0 of 14 mice (0%) receiving 30 mg/kg TNP-470. However, TNP-470 had no effect on the tumor growth. These results indicate that the angiogenesis inhibitor TNP-470 has a strong inhibitory activity against in vivo liver metastasis of human gastric carcinoma.

Topics: Animals; Antibiotics, Antineoplastic; Cell Division; Cyclohexanes; Female; Humans; Liver Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Mitomycin; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Stomach Neoplasms; Tumor Cells, Cultured

The ability of the angiogenesis inhibitor TNP-470 to prevent liver metastasis after partial hepatectomy, and whether TNP-470 impairs liver regeneration or skin wound healing, was evaluated. Following the injection of VX2 carcinoma cells into the portal vein of rabbits, half of the animals underwent resection of the middle hepatic lobe (hepatectomized group) and half did not (non-hepatectomized group). TNP-470 (50 mg) was infused continuously into the portal vein in both groups for 7 days, while controls received only water. The hepatectomized TNP-470-treated group had significantly fewer tumours (mean(s.e.m.) 23.3(12.3)) than the hepatectomized control group (123.7(24.4)). There was no significant difference in the 5-bromo-2'-deoxyuridine labelling index of regenerated hepatocytes between the TNP-470-treated and control groups. Wound healing in TNP-470-treated animals was not impaired. Intraportal infusion of TNP-470 prevents the recurrence of liver metastasis after partial hepatectomy without impairing healing or liver regeneration.

Topics: Animals; Cyclohexanes; Hepatectomy; Liver Neoplasms; Neoplasm Seeding; O-(Chloroacetylcarbamoyl)fumagillol; Rabbits; Sesquiterpenes; Wound Healing

Angiogenesis inhibitors have attracted considerable interest. The anti-tumor and anti-metastatic effects of TNP-470, an angiogenesis inhibitor, and mitomycin C (MMC), a representative anti-neoplastic agent, were investigated using a xenotransplanted human colon cancer, TK-4. Suturing of small pieces of TK-4 tumors to the cecal wall in nude mice (orthotopic transplantation) induced liver metastasis. Mice were randomly divided into 3 groups; a control group given saline solution, a group receiving TNP-470 and a group receiving MMC. TNP-470 was given s.c. on alternate days for 5 weeks from day 10 after cecal transplantation and MMC was administered intraperitoneally (i.p.) once a week from day 10 after cecal transplantation. MMC significantly inhibited cecal tumor growth. In the control group, liver metastases developed in 9 out of 10 mice, including 3 with more than 20 metastatic foci. Liver metastasis also developed in 8 out of 10 mice receiving MMC, 2 of which had many metastases. In contrast, liver metastasis developed in only 2 out of 8 mice in the TNP-470 group and neither of these animals had numerous metastases.

Topics: Animals; Antibiotics, Antineoplastic; Body Weight; Cell Division; Colonic Neoplasms; Cyclohexanes; Humans; Liver Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Mitomycin; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes

We investigated the effects of an angiogenesis inhibitor, TNP-470, in suppressing the recurrence in the liver remnant after partial hepatectomy for VX2 liver metastases model of rabbits. After inoculation of 1 x 10(6) VX2 tumor cells via the portal vein, the medial lobe of the liver was resected. Then, TNP-470 (7 mg/body/day) was infused (Group 1), or distilled water (Group 2), continuously via the mesenteric vein for a week. The control group received only infused distilled water via the mesenteric vein. Fourteen days after the tumor inoculation, there were 66.0 +/- 20.2 (mean +/- SD) metastatic colonies on the liver surface of control group, against 23.3 +/- 24.0 and 123.7 +/- 47.9 colonies in Group 1 and Group 2, respectively. There were significant differences between control group and Group 2, and between Group 1 and Group 2. We consider that the liver regeneration following hepatectomy might provoke tumor growth of occult metastases in the liver remnants. TNP-470 suppressed these growth by its anti-neovascular activity. And this effect reduced the number of metastatic colonies. Fourteen days after hepatectomy, the weights of the liver were no different among 3 groups. The BrdU Labeling Index was also no different between Group 1 and Group 2. TNP-470 did not suppress the regeneration of the liver remnant. We consider that the systemic side effects of TNP-470 might be made minimal, because this agent affects only on endothelial cells. These results suggest that the angiogenesis inhibitor TNP-470 is beneficial to suppress hepatic recurrence after partial hepatectomy for liver metastases without suppression of liver regeneration.

Topics: Animals; Antibiotics, Antineoplastic; Cyclohexanes; Hepatectomy; Liver Neoplasms; Liver Regeneration; Male; Neoplasm Recurrence, Local; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rabbits; Sesquiterpenes