o-(chloroacetylcarbamoyl)fumagillol and Colorectal-Neoplasms

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

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 timing and length of administration of angiogenesis inhibitor TNP-470 was altered to evaluate the effect on disease progression in a rat model of colorectal hepatic metastases.. Pair-fed BD-IX rats, injected intrasplenically with rat colon adenocarcinoma K12/TRb cells at day 0, were randomized to receive subcutaneous injections of either placebo or 15 mg/kg TNP-470 on alternate days: for 2 weeks beginning 24 hours after tumor inoculation ("Early"), for 4 weeks beginning 24 hours after tumor inoculation ("Prolonged"), or for 2 weeks beginning at day 15 after macroscopic tumor nodules were confirmed ("Delayed"). Response to treatment was evaluated by counting tumor nodules on the surface of the liver at laparotomy on day 14 and 28 after tumor inoculation. The animals were followed for survival and cause of death.. Maximal suppression of hepatic metastases at day 28 required 4-week rather than 2-week TNP-470 administration. Prolonged TNP-470 administration resulted in significantly fewer hepatic metastases at day 28 compared to control (P < .05). Early and prolonged TNP-470 improved survival (Wilcoxon test, P < .05) compared with delayed TNP-470 and placebo. Delayed TNP-470 administration did not increase survival or significantly diminish the number of metastases at day 28 compared with placebo.. These data suggest that prolonged adjuvant antiangiogenic therapy may suppress colorectal hepatic micrometastases.

Topics: Animals; Antibiotics, Antineoplastic; Colorectal Neoplasms; Cyclohexanes; Liver Neoplasms, Experimental; Male; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Sesquiterpenes; Time Factors; 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