o-(chloroacetylcarbamoyl)fumagillol and Colonic-Neoplasms

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

The angiogenesis inhibitor TNP-470 (AGM-1470) has shown encouraging results in animal models of established tumors. However, results of recent clinical trials using TNP-470 have been disappointing. Since little is known about the effects of TNP-470 at the minimal disease stage, we analyzed the effects of TNP-470 on the early stages of tumor establishment.. Twenty thousand green fluorescent protein (GFP)-transfected murine CT-26 (colonic carcinoma) or Panc-02-H0 (pancreatic adenocarcinoma) cells were inoculated in dorsal skin-fold chambers in BALB/c or C57BL6 mice. Tumor area and microvessel density (MVD) were quantified by intravital microscopy (IVM). Body weight was also monitored. Effects were compared with those in a conventional model involving subcutaneous (s.c.) inoculation of 10(6) tumor cells, followed by measurement of tumor volume, endogenous plasma VEGF/endostatin (ELISA) and proliferation/apoptosis/microvessel density (Ki-67/TUNEL/CD-34). TNP-470 was injected s.c. over the 10-day experimental period (30 mg/kg every 2 days [n=6] to 100 mg/kg/day [n=5 dorsal skin-fold chamber model, n=4 s.c. tumor model]).. At 30 mg/kg/every second day neither CT-26 nor PANC-02-H0 tumors were inhibited in neither of the two models. TNP-470 dosage was escalated in CT-26-bearing animals until an antiangiogenic effect could be observed. In the IVM model, only TNP-470 100 mg/kg/day reduced MVD (P=0.006), but failed to block the onset of angiogenesis and tumor area increase. Body weight decreased by 25% (P<0.05). In the subcutaneous tumor model, tumor growth was reduced (P=0.045) but not blocked, while vascular endothelial growth factor (VEGF)/endostatin synthesis and Ki67/TUNEL/CD-34 were not significantly affected.. While capable of reducing tumor growth in a conventional model, treatment with TNP-470 does not block the onset of angiogenesis and tumor establishment in a model of minimal disease. When used as a single agent TNP-470 does not control minimal tumor disease in experimental colonic carcinoma.

Topics: Angiogenesis Inhibitors; Animals; Carcinoma; Cell Proliferation; Colonic Neoplasms; Cyclohexanes; Endothelium, Vascular; Humans; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Neoplasms, Experimental; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Pancreatic Neoplasms; Sesquiterpenes; Treatment Failure; Vascular Endothelial Growth Factor A

We investigated the potentiation of combination therapy using tumor necrosis factor (TNF) with TNP-470, a potent angiogenesis inhibitor.. We evaluated the antitumor effect in vivo against subcutaneous (s.c.) MC38 mouse colon adenocarcinoma tumors in C57BL/6 mice. The mice were treated with a single bolus injection via the tail vein of 3 or 8 microg rhTNF in 0.5% bovine serum albumin/normal saline (BSA/NS), or with 0.5% BSA/NS alone as a control, with or without TNP-470 pretreatment, given as 30 or 60 mg/kg x 2 days, s.c. DNA synthesis in human umbilical endothelial cells (HUVEC) was assessed by [(3)H]thymidine uptake after incubation with TNF, with or without TNP-470.. The antitumor effect of TNP-470 pretreatment combined with 3 microg recombinant human (rh) TNF injection resulted in an 80% reduction of tumor volume compared with the control. This was significantly better than that induced by 3 microg rhTNF alone (P < 0.005). DNA synthesis in HUVEC was inhibited by TNF with TNP-470 in a dose-dependent manner, but there was no enhanced effect against MC38 in vitro.. These results suggest that the combination of the angiogenesis inhibitor TNP-470 and TNF might have a synergistic antitumor effect on solid tumors in vivo.

Topics: Adenocarcinoma; Angiogenesis Inhibitors; Animals; Cell Line, Tumor; Colonic Neoplasms; Cyclohexanes; DNA, Neoplasm; Endothelium, Vascular; Mice; Mice, Inbred C57BL; Neoplasms, Experimental; O-(Chloroacetylcarbamoyl)fumagillol; Recombinant Proteins; Sesquiterpenes; Treatment Outcome; Tumor Necrosis Factor-alpha; Umbilical Veins

To study the effect of TNP-470 on cell growth, proliferation and apoptosis in human colon cancer xenografts in nude mice.. Human colon cancer xenografts were transplanted into 20 nude mice. Mice were randomly divided into two groups. TNP-470 treated group received TNP-470(30 mg/kg, s.c) every other day and the control group received a sham injection of same volume saline solution. They were sacrificed after 4 weeks and their tumors were processed for histological examination. The expression of proliferating cell nuclear antigen (PCNA) in tumors was detected using immunohistochemical method with image analysis, and apoptosis in tumor cells was measured by TdT-mediated biotinyated-dUTP nick end labeling (TUNEL) staining.. Comparing with controls, tumor growth was significantly inhibited in TNP-470 treated group, the inhibitory rate being 54.4 %. Expression of PCNA in tumors of TNP-470 treated group (PI 54.32+/-11.47) was significantly lower than that of control group (PI 88.54+/-12.36), P<0.01. Apoptosis index (AI) of TNP-470 treated group (18.95+/-1.71) was significantly higher than that of control group (7.26+/-1.44), P<0.001, typical morphological change of apoptosis in tumor cells was observed in TNP-470 treated group.. Besides the anti-angiogenic effects, TNP-470 can inhibit tumor growth by inhibiting the proliferation and inducing apoptosis of tumor cells.

Topics: Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Apoptosis; Cell Division; Colonic Neoplasms; Cyclohexanes; Mice; Mice, Nude; Neoplasm Transplantation; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Transplantation, Heterologous

To investigate antitumour efficacy of the combination of the antiangiogenic agent TNP-470 combined with chemoimmunotherapy in different tumour models in mice. B6D2F1 mice and BALB/c mice were inoculated in the footpad of the right hind limb with B16F10 melanoma cells or colon adenocarcinoma cells C-26, respectively. Subsequently, they received therapy consisting of TNP-470 and/or IL-12 and tumour growth was observed. In the melanoma model this therapy regimen was combined with cisplatin in a subtherapeutic dose. The antiangiogenic action of the tested agents was evaluated using tumour-induced angiogenesis assay in vivo. In order to analyse interactions between TNP-470 (or cisplatin) and IFN-gamma on tumour cells in vitro, the following methods were used: MTT assay, Western blot analysis, and flow cytometry analysis.. Administration of the combined therapy with TNP-470 and IL-12 resulted in augmented antitumour activity in colon-26 and B16F10 melanoma models. Addition of cisplatin further enhanced efficacy of this combined therapy in the melanoma model. We showed that antitumour activity of this combined therapy is mediated by multiple mechanisms: not only is enhancement of the antiangiogenic activity mediated by TNP-470 and IL-12 but also by the synergistic cytostatic/cytotoxic action of IL-12-induced IFN-gamma and TNP-470 or cisplatin on tumour cells. The experiments revealed that TNP-470 together with IFN-gamma leads to the increased expression of p21 protein in cancer cells, which in turn may contribute to their cytostatic/cytotoxic action in vitro.. Our experiments show a successful TNP-470-based combination therapy and suggest that the enhancement of the antitumour activity could be explained by a concomitant effect on both endothelial and tumour cell compartments.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Benzothiazoles; Blotting, Western; Cell Division; Cisplatin; Colonic Neoplasms; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Cyclohexanes; Drug Synergism; Drug Therapy, Combination; Flow Cytometry; Immunotherapy; Interferon-gamma; Interleukin-12; Melanoma, Experimental; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred DBA; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Tetrazolium Salts; Thiazoles; Toluene; Tumor Suppressor Protein p53

To study the effect of angiogenesis inhibitor TNP-470 on peritoneal dissemination of colon cancer in nude mice.. The MTT assay was used to evaluate the inhibitory effect of TNP-470 on human colon cancer cell line Lovo. Lovo cells were injected into the peritoneal cavity of BABL/C nu/nu mice and the models of peritoneal dissemination were developed. Thirty nude mice were randomly divided into control and TNP-470-treated group. In TNP-470-treated group, TNP-470 was injected subcutaneously every other day from day 1 until sacrifice or death (30 mg x kg(-1)). The control group received a sham injection of the same volume saline solution.. In vitro, TNP-470 inhibited the growth of Lovo cells, with its IC50 at 2.14 X 10(2) microg x L(-1). In vitro, TNP-470 demonstrated growth inhibition of tumors. Mice body weight and abdominal circumferences were significantly different between TNP-470-treated group (24.5+/-3.2 g, 7.0+/-1.1 cm) and control group (29.5+/-2.1 g, 10.3+/-1.5 cm), P=0.005 and P=0.001. The number of disseminated foci was significantly different between the control group (92.1+/-20.6) and the TNP-470-treated group (40.3+/-12.3), P<0.001. The maximal size of foci was significantly smaller in TNP-470-treated group (3.3+/-0.7 mm) than that of control (7.3+/-2.3 mm), P=0.004. Mean survival time was significantly longer in TNP-470-treated group(98.00+/-12.06 d) than that in control group (41.86+/-9.51 d), P<0.001.. Angiogenesis inhibitor TNP-470 might be effective in treating peritoneal dissemination of colon cancer and improve the survival rate of nude mice.

Topics: Adenocarcinoma; Angiogenesis Inhibitors; Animals; Cell Division; Colonic Neoplasms; Cyclohexanes; Female; Humans; In Vitro Techniques; Mice; Mice, Inbred BALB C; Mice, Nude; O-(Chloroacetylcarbamoyl)fumagillol; Peritoneum; Sesquiterpenes; Tumor Cells, Cultured

Neovascularization is critical for growth of primary tumors and their distant metastasis. This study was designed to investigate the effects of angiogenesis inhibitor TNP-470 in combination with 5-fluorouracil (5-FU) on the growth of human colon cancer cell line, LOVO, in vitro and in vivo.. In vitro, the growth inhibitory effect of TNP-470 or 5-FU on LOVO cells was examined by MTT assay, and synergetic effect of these two agents was evaluated by Burgi analysis. In vivo, human colon cancer xenografts were transplanted into BALB/C nude mice and treated with TNP-470 alone and combination with 5-FU. Tumor microvessel density (MVD) was measured by immunostaining with anti-von Willebrand factor monoclonal antibody; the expression of vascular endothelial growth factor (VEGF) in tumor tissue was detected using an immunohistochemical method with image analysis system.. In vitro, TNP-470 and 5-FU inhibited the growth of LOVO cells, with the IC50 at 55.8 ng/ml and 4.6 ng/ml, respectively. Burgi analysis revealed synergism of these two agents, a remarkable growth inhibitory effect on LOVO cells was obtained. In vivo, in every treatment group, tumor growth was suppressed significantly. The combination group showed significant enhancement in antitumor efficacy. Expression of VEGF in tumors was clearly inhibited by TNP-470 in combination with 5-FU or 5-FU alone compared to the control. Tumor MVD was lower in TNP-470 combination with 5-FU group or TNP-470 group compared to control group.. TNP-470 suppressed the growth of LOVO cells and human colon cancer xenografts and the angiogenesis; TNP-470 in combination with 5-FU might product synergetic effect.

Topics: Animals; Body Weight; Cell Division; Colonic Neoplasms; Cyclohexanes; Disease Models, Animal; Drug Combinations; Endothelial Growth Factors; Female; Fluorouracil; Humans; Intercellular Signaling Peptides and Proteins; Lymphokines; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Neoplasms, Experimental; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors; Xenograft Model Antitumor Assays

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

We have investigated the antimetastatic effect of TNP-470 against postoperative lung metastasis following the removal of human colon cancer xenotransplanted into nude rat. The KM12SM human colon cells were injected into the cecal wall. At 5 weeks after the injection, the cecum was removed including the tumor. Then, TNP-470 was administered continuously by subcutaneous injection pump at a dosage of 30 mg/kg/week. The Control Group received no administration of TNP-470. Group A and Group B received administration of TNP-470 just after the cecal removal for 4 and 2 weeks, respectively. Group C received 2 weeks' administration of TNP-470 from week 3 after the removal. The survival rate of each group was calculated, and any lung metastasis was evaluated macro and microscopically. At 7 weeks after the removal, lung metastasis was detected in all rats of the Control Group, and in 4 of 8 rats in Group C. In Groups A and Group B, only one rat developed lung metastasis. The 30 week-survival rate in Group A and that in Group B was significantly higher than that in the Control Group or Group C. Moreover, the incidence of lung metastasis at the time of death or 30 weeks after the removal in Group A, and in Group B, was lower than that in Control Group or Group C. The angiogenesis inhibitor, TNP-470 showed an excellent antimetastatic effect against postoperative lung metastasis from transplanted human colon cancer.

Topics: Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Colonic Neoplasms; Cyclohexanes; Lung Neoplasms; Male; Neoplasm Transplantation; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Rats, Inbred F344; Sesquiterpenes; Survival Rate; Transplantation, Heterologous; Tumor Cells, Cultured

The present study was designed to evaluate the effects of perioperative treatment with TNP-470 on the resistance of colonic anastomoses. TNP-470 is a drug that was developed as an inhibitor of angiogenesis.. A colonic anastomosis was constructed in Sprague-Dawley rats. From 4 days before surgery to 4 days afterwards, each animal received daily intraperitoneal treatment that differed according to the group to which it belonged: the control group, which received gum arabic, or the TNP group, which received 30 mg/kg TNP-470 in gum arabic. The size of the cecum and the diameters of the pre-anastomotic and post-anastomotic colon were measured at operation and 4 days after surgery, when all animals were sacrificed. At this time the presence of adhesions was also investigated. Each segment containing an anastomosis was removed and the bursting pressure (BP) and bursting wall tension (BWT) were determined.. Loss of cecum caliber and decreases in pre-anastomotic diameter were significantly greater in the TNP group than in the control group (p = 0.017 and p = 0.004, respectively). Dilatation and obstruction of the colon were more frequent in the control group, but the difference between the groups was significant only with respect to dilatation (p = 0.005). Moreover, loss of body weight was greater in the TNP group than in the control group (p < 0.001). BP and BWT were significantly lower in animals that had received TNP-470 (p = 0.04 and p = 0.005, respectively). With respect to post-anastomotic diameter, general adhesions and adhesions to the anastomotic line, there were no statistically significant differences between the groups.. Perioperative treatment with TNP-470, an inhibitor of angiogenesis, affects the healing and reduces the resistance of colonic anastomoses.

Topics: Anastomosis, Surgical; Angiogenesis Inhibitors; Animals; Colon; Colonic Neoplasms; Cyclohexanes; Female; Male; O-(Chloroacetylcarbamoyl)fumagillol; Perioperative Care; Prospective Studies; Random Allocation; Rats; Rats, Sprague-Dawley; Sesquiterpenes; Wound Healing

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 antimetastatic effect of a potent angiogenesis inhibitor, O-(chloroacetyl-carbamoyl)fumagillol (TNP-470), was investigated in nude mice implanted with human colon cancer. Small pieces of tumors from three established human colon cancer cell lines (TK-3, TK-4, and TK-9), which were maintained in nude mice, were implanted into the cecal wall of nude mice via a small incision in the serosa. TNP-470 (20 or 30 mg/kg) was given s.c. every other day from day 10 after implantation, and the mice were sacrificed after 6 weeks. There was no difference in the weight of the implanted tumors (control group: 0.45 +/- 0.29 g versus treated group: 0.49 +/- 0.27 g). An antimetastatic effect of TNP-470 was clearly demonstrated in a dose-dependent manner. In the mice given 20 mg/kg TNP-470, liver metastasis developed in 3 of 10 cases. In the 30-mg/kg group, metastasis developed in only 1 of 17 mice, while it developed in 22 of 32 mice of the control group. The number of metastatic foci was significantly less in the treated groups. TNP-470 effectively prevented liver metastasis, however, but had no effect on the growth of the primary tumor. These results indicate that the angiogenesis inhibitor TNP-470 has a strong inhibitory activity against in vivo hepatic metastasis of human colon cancer.

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

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

Angiogenesis is a fundamental process by which new blood vessels are formed. Progressive tumor growth necessitates the continuous induction of new capillary blood vessels which converge upon the tumor. Suppression of tumor growth can be accomplished with the use of antiangiogenesis agents. AGM-1470 is a potent angiogenesis inhibitor in vitro and in vivo. In mouse studies, AGM-1470 has suppressed the growth and neovascularization induced by four murine tumors resulting in a 55% to 77% decrease in tumor growth. In these mice significant toxicity did not result from AGM-1470 therapy. AGM-1470 administered systemically to C57BI/6 male mice for 20 to 28 days inhibited the growth of: (1) Lewis lung carcinoma resulting in a T/C (treatment/control = mean tumor volume of treated/mean tumor volume of control) of 0.38 +/- 0.03 (P < .001); (2) colon adenocarcinoma 38 resulting in a T/C of 0.23 +/- 0.02 (P < .001); and (3) fibrosarcoma 105 resulting in a T/C of 0.31 +/- 0.05 (P < .001). To determine if antiangiogenic therapy was equally effective in mice of both sexes and in immunodeficient animals, we tested AGM-1470 in the treatment of fibrosarcoma 105 in both female mice and nude mice. For female mice T/C was 0.24 +/- 0.06 (P < .001). For nude mice T/C was 0.27 +/- 0.06 (P < .001). These results demonstrate that AGM-1470 suppresses the growth of a variety of different tumors. Furthermore, the antitumor effect of AGM-1470 therapy is independent of the immune system and sex.

Topics: Adenocarcinoma; Animals; Antibiotics, Antineoplastic; Body Weight; Colon; Colonic Neoplasms; Cyclohexanes; Female; Fibrosarcoma; Immunocompromised Host; Lung; Lung Neoplasms; Male; Mice; Mice, Nude; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Sex Factors