o-(chloroacetylcarbamoyl)fumagillol and Carcinoma--Hepatocellular

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

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

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

Effective therapy for advanced hepatocellular carcinoma (HCC) is lacking. Conventional chemotherapy was judged to be ineffective. We previously demonstrated that the histone deacetylase inhibitor Trichostatin A (TSA) blocks growth of HCC cells in vitro. The anti-tumoral effect of a combination of more than 2 classes of drugs remains unexplored. Four hepatoma cell lines were incubated with increasing concentrations of Tamoxifen (TAM), 9-cis retinoic acid (CRA), the methioninaminopeptidase inhibitor TNP-470 and TSA as single agents and in combination. Anti-proliferative and pro-apoptotic effects were assessed using BrdU-incorporation, FACS analysis and immunocytochemistry. Central pro- and anti-apoptotic proteins were measured by semi-quantitative Western blotting and substrate assays. All single substances inhibited proliferation and induced apoptosis in HCC cells only at high concentrations. The combination of TAM/CRA/TNP/TSA multiplied the anti-tumoral effects, reaching up to 93% inhibition of proliferation and 63% induction of apoptosis after 24 h in Hep1B cells. Pro-apoptotic factors bax and caspase 3 were highly increased with quadruple therapy, while anti-apoptotic bcl-2 decreased to undetectable levels. Fibroblasts remained largely unaffected. While the single substances were not effective on hepatoma cells in tolerable doses, their combination significantly increases anti-tumoral efficacy. Combination therapy with biomodulators is a promising treatment option for HCC.

Topics: Animals; Antineoplastic Agents; Apoptosis; bcl-2-Associated X Protein; Carcinoma, Hepatocellular; Caspase 3; Caspases; Cell Cycle; Cell Division; Cell Line, Tumor; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Cyclohexanes; DNA; Drug Synergism; Enzyme Activation; Enzyme Inhibitors; Estrogen Antagonists; Humans; Hydroxamic Acids; Immunologic Factors; Mice; O-(Chloroacetylcarbamoyl)fumagillol; Proto-Oncogene Proteins c-bcl-2; Rats; Sesquiterpenes; Tamoxifen; Tretinoin

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

We evaluated the effects of angiogenesis inhibitor, TNP-470, on hepatocellular carcinoma (HCCs) induced by a choline-deficient L-amino acid defined (CDAA) diet in rats. Male Fisher 344 rats were fed CDAA for 68 weeks. Rats were treated by subcutaneous injection of TNP-470 (15 mg/kg) or saline (control) three times per week from 53 to 68 weeks. At the end of the experiment, we determined the frequency and size of HCCs and glutathione S-transferase placental form (GSTP)-positive lesions, histology of liver cirrhosis, liver function, and liver weight per body weight. We also determined, using histologic and immunohistochemical semiquantification analyses, the degree of vascularity, apoptosis and proliferation in HCC and adjacent tissues. Treatment with TNP-470 resulted in a reduction of the size and frequency of HCC compared to untreated rats. However, TNP-470 did not influence the histology of liver cirrhosis and liver function. The liver weight per body weight of TNP-470-treated rats was slightly heavier in comparison with that of the controls. Treatment with TNP-470 significantly reduced tumor vascularity relative to the controls. There were no significant differences in the Ki-67 labeling index of HCCs between TNP-470 treated and control rats. The frequency of apoptotic hepatoma cells in TNP-470-treated rats was higher than in control rats. Our results indicate that TNP-470 suppresses the progression of CDAA-induced HCCs in rats through inhibition of angiogenesis, and suggest that TNP-470 might be useful clinically for HCCs.

Topics: Angiogenesis Inhibitors; Animals; Apoptosis; Carcinoma, Hepatocellular; Choline; Cyclohexanes; Diet; Fatty Acids, Unsaturated; Liver Neoplasms, Experimental; Lung Neoplasms; Male; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Rats, Inbred F344; Sesquiterpenes

Mechanisms regulating angiogenesis in human hepatomas were analyzed. Huh7 hepatoma cells transplanted into athymic mice formed highly vascularized reddish tumors with abundant microvessels, while angiogenesis by PLC/PRF/5 hepatoma cells was less remarkable. However, the production of angiogenesis stimulators such as VEGF and IL6 by Huh7 cells was much less than by PLC/PRF/5 cells. In addition, the production of angiogenesis inhibitor TSP-1 by Huh7 cells was greater than by PLC/PRF/5 cells. Therefore, the difference in angiogenesis between these two hepatomas was not explained by the production of these known angiogenesis regulators. On the other hand, PLC/PRF/5 cells but not Huh7 cells secreted an inhibitor of the proliferation of vascular endothelial cells, which was enhanced by p53-gene transfer. These results suggest that the production of this p53-inducible angiogenesis inhibitor is responsible, at least partly, for the regulation of angiogenesis in human hepatomas.

Topics: Animals; Antibiotics, Antineoplastic; Body Weight; Carcinoma, Hepatocellular; Cell Division; Culture Media, Conditioned; Cyclohexanes; Cytokines; Endothelium, Vascular; Gene Transfer Techniques; Humans; Immunohistochemistry; Mice; Mice, Inbred ICR; Mice, Nude; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Thrombospondin 1; Tumor Cells, Cultured; Tumor Suppressor Protein p53

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

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