o-(chloroacetylcarbamoyl)fumagillol and Adenocarcinoma

Aging is considered one of the main predisposing factors for the development of prostate malignancies. Angiogenesis is fundamental for tumor growth and its inhibition represents a promising therapeutic approach in cancer treatment. Thus, we sought to determine angiogenic responses and the effects of antiangiogenic therapy in the mouse prostate during late life, comparing these findings with the prostatic microenvironment in the Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) model.. Male mice (52 week-old FVB) were submitted to treatments with SU5416 (6 mg/kg; i.p.) and/or TNP-470 (15 mg/kg; s.c.). Finasteride was administered (20 mg/kg; s.c.), alone or in association to both inhibitors. The dorsolateral prostate was collected for VEGF, HIF-1α, FGF-2 and endostatin immunohistochemical and Western Blotting analyses and for microvessel density (MVD) count.. Senescence led to increased MVD and VEGF, HIF-1α and FGF-2 protein levels in the prostatic microenvironment, similarly to what was observed in TRAMP mice prostate. The angiogenic process was impaired in all the treated groups, demonstrating significantly decreased MVD. Antiangiogenic and/or finasteride treatments resulted in decreased VEGF and HIF-1α levels, especially following TNP-470 administration, either alone or associated to SU5416. The combination of these agents resulted in increased endostatin levels, regardless of the presence of finasteride.. Prostatic angiogenesis stimulation during senescence favored the development of neoplastic lesions, considering the pro-angiogenic microenvironment as a common aspect also observed during cancer progression in TRAMP mice. The combined antiangiogenic therapy was more efficient, leading to enhanced imbalance towards angiogenic inhibition in the organ. Finally, finasteride administration might secondarily upregulate the expression of pro-angiogenic factors, pointing to the harmful effects of this therapy.

Topics: 5-alpha Reductase Inhibitors; Adenocarcinoma; Angiogenesis Inhibitors; Animals; Blotting, Western; Cyclohexanes; Disease Models, Animal; Endostatins; Fibroblast Growth Factor 2; Finasteride; Hypoxia-Inducible Factor 1, alpha Subunit; Immunohistochemistry; Indoles; Male; Mice; Mice, Transgenic; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Prostatic Neoplasms; Pyrroles; Sesquiterpenes; Tumor Microenvironment; Vascular Endothelial Growth Factor A

Angiogenesis is required for solid tumor growth and facilitates tumor progression and metastasis. The inhibition effects of O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), an angiogenesis inhibitor, and gemcitabine, a chemotherapeutic agent, on expression of growth factors were investigated using human pulmonary adenocarcinoma cell line, A549. The A549 cells were divided into four groups: control group, 10(-6) mg/ml gemcitabine treated group, 10(-4) mg/ml TNP-470 treated group and gemcitabine+TNP-470 treated group. The mRNA and protein expression of vascular endothelial growth factor (VEGF) and its receptors, FMS-like tyrosine kinase-1 (FLT-1) and kinase insert domain-containing receptor (KDR), in different groups were measured. The growth of A549 cell cultured with gemcitabine or TNP-470 was inhibited in an almost dose-dependent manner. Although gemcitabine (10(-6) mg/ml) alone and TNP-470 (10(-4) mg/ml) alone had no effect on the mRNA and protein expression of VEGF and its receptors (FLT-1, KDR) in A549 cells compared to the control (P>0.05), 10(-6) mg/ml gemcitabine in combination with 10(-4) mg/ml TNP-470 had significant effect (P<0.01). Moreover, combination of the two drugs significantly inhibited the mRNA expression of VEGF, FLT-1 and KDR compared to either drug alone (P<0.05). This study suggests that combined treatment with TNP-470 plus gemcitabine may augment the antiangiogenic and antineoplastic effects in lung cancer cells in vitro.

Topics: Adenocarcinoma; Antineoplastic Combined Chemotherapy Protocols; Cell Line, Tumor; Cell Proliferation; Cyclohexanes; Deoxycytidine; Disease Progression; Gemcitabine; Gene Expression Regulation, Neoplastic; Humans; Lung Neoplasms; Neoplasm Metastasis; O-(Chloroacetylcarbamoyl)fumagillol; Protein Structure, Tertiary; Sesquiterpenes; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1; Vascular Endothelial Growth Factor Receptor-2

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

Most cases of pancreatic cancer are inoperable when diagnosed. Since immunotherapy and antiangiogenic therapy have been reported to be promising for pancreatic cancer, we examined whether the combination of immunotherapy with dendritic cells (DCs) and the antiangiogenic drug TNP-470 induces tumor regression. Syngeneic mouse pancreatic adenocarcinoma cells were orthotopically inoculated into C57/BL6 mice. DCs with or without tumor lysate (TL) were administered i.p. at 4 and 5 weeks. TNP-470 was injected s.c. into tumor-bearing mice every other day from 4 weeks to 6 weeks. We compared anticancer effects in 6 groups: NT (no treatment), DC/TL- (DCs without TL), DC/TL+ (DCs pulsed with TL), TNP (TNP-470 alone), DC/TL-TNP (DC/TL- plus TNP-470) and DC/TL+TNP (DC/TL+ plus TNP-470). We measured tumor volume, mean vascular density (MVD) and vessel diameter by FITC-dextran using an intravital microscope; degrees of proliferation and apoptosis of cancer cells by PCNA and TUNEL; infiltrating lymphocytes and expression levels of VEGF and MMP-9 by immunohistochemistry and immunoblotting. Tumor volume and MVD were significantly suppressed in the treatment groups with prolonged survival rate, especially in the DC/TL+TNP group. There were no significant differences in apoptosis among the 6 groups except DC/TL+. The number of infiltrating CD4+ cells in the DC/TL+ group was higher than that in the NT group. VEGF expression was significantly suppressed in the treatment groups containing TNP-470, and MMP-9 was also suppressed in the groups containing DC/TL+. Our data suggested that TL-pulsed DCs combined with TNP-470 induced regression of mouse pancreatic cancer, possibly through induction of immune responses and suppression of angiogenesis.

Topics: Adenocarcinoma; Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Cell Division; Cell Survival; Combined Modality Therapy; Cyclohexanes; Dendritic Cells; Disease Models, Animal; Immunotherapy; Lymphocyte Activation; Lymphocyte Transfusion; Mice; O-(Chloroacetylcarbamoyl)fumagillol; Pancreatic Neoplasms; Sesquiterpenes; Spleen

Treating tumor with angiogenesis inhibitor and chemotherapeutic drugs is a research hot spot now. This study was designed to observe the synergetic inhibitory effect of fumagillol (TNP-470) in combination with cyclophosphamide (CTX) on metastasis of lung adenocarcinoma cell line LA795 xenograft in mouse, and to explore the related mechanism of suppressing tumor metastasis by TNP-470.. Forty T739 nude mice bearing highly metastatic LA795 cells were randomized into 5 groups: control group, vehicle group, TNP-470 (30 mg/kg) group, CTX (40 mg/kg) group, and combination group (TNP-470 plus CTX). All mice were killed 3 weeks laterû the subcutaneous tumors were weighted to calculate inhibitory rate. The metastatic tumor foci on lung surface in mice were counted to calculate occurrence rate and inhibitory rate of metastases on lung surface. The microvessel density (MVD) and the expression of tumor metastasis-related factor P-selectin in subcutaneous tumor were detected by immunohistochemistry and analyzed with image analysis system.. The inhibitory rate of tumor was significantly higher in combination group (81.5%) than in other groups (P<0.01). TNP-470 plus CTX showed synergetic effect on inhibiting metastasis on lung surface with a Q value of 1.21. The metastatic foci on lung surface were significantly fewer in combination group, TNP-470 group, and CTX group than in control group (1.75+/-1.71, 4.75+/-3.34, and 8.50+/-2.67 vs. 12.13+/-4.02, P<0.05). The MVD and the expression of P-selectin in subcutaneous tumor were also significantly lower in combination group and TNP-470 group than in control group (9.13+/-1.61 and 12.13+/-2.84 vs. 20.50+/-3.12, P<0.01; 5.25+/-2.27 and 7.13+/-3.01 vs. 13.75+/-3.38, P<0.01).. TNP-470 and CTX have synergetic inhibitory effect on lung metastasis of LA795 xenograft tumor. TNP-470 may inhibit lung metastasis of LA795 xenograft tumor by suppressing the expression of P-selectin.

Topics: Adenocarcinoma; Angiogenesis Inhibitors; Animals; Antineoplastic Agents, Alkylating; Cell Line, Tumor; Cyclohexanes; Cyclophosphamide; Drug Synergism; Lung; Lung Neoplasms; Male; Mice; Mice, Nude; Microcirculation; Neoplasm Metastasis; O-(Chloroacetylcarbamoyl)fumagillol; P-Selectin; Random Allocation; Sesquiterpenes

To elucidate the efficacy of TNP-470 on ovarian carcinomas by using cis-diamminedichloroplatinum (CDDP)-resistant cell lines.. The susceptibility of human ovarian carcinoma-derived cell lines and its resistant cell lines against cis-diamminedichloroplatinum (CDDP) to angiogenesis inhibitor, TNP-470, were analyzed using three human cultured cell lines derived from ovarian carcinoma (TYK, KF-92, and Nakajima) and each CDDP-resistant cell line (TYK-R, TYK-R', KF/ra, KF/rb, Nakajima-S1, and Nakajima-S2).. TNP-470 revealed suppression of thymidine incorporation by all of the nine cell lines linearly dependent on the concentration of TNP-470. Significant suppression was not observed for either uridine or leucine incorporation by all nine cell lines. To elucidate the site of each cell line, in which TNP-470 revealed the antitumor effect, the incorporation of (3)H-TNP-470 by cultured cells or by DNA extracted from cultured cells was examined in the cell lines, and the ratio of (3)H-TNP-470 incorporation by DNA to (3)H-TNP-470 incorporation by cultured cells ranged from 2.3% to 4.4% in three parent cell lines. The ratio in the CDDP-resistant cell lines ranged from 11.0% to 46.7%. The ability of TNP-470 to inhibit neoplastic growth in vivo was evaluated using KF-92, KF/ra, KF/rb, Nakajima, Nakajima-S1, and Nakajima-S2. Concerning KF-92, KF/ra, and KF/rb, 30 mg/kg of TNP-470 significantly suppressed the tumorigenicity of KF-92, 10 and 30 mg/kg of TNP-470 suppressed the tumorigenicity of KF/ra, and 30 mg/kg of TNP-470 suppressed the tumorigenicity of KF/rb. Concerning Nakajima, Nakajima-S1, and Nakajima-S2, TNP-470 revealed no inhibitory effect on the tumorigenicity of Nakajima. Contrary, significant inhibition was observed when 30 mg/kg of TNP-470 was used to the CDDP-resistant cell lines Nakajima-S1 and Nakajima-S2.. These results suggest that the clinical application of TNP-470 may be one of the possible treatments for the CDDP-resistant ovarian carcinomas.

Topics: Adenocarcinoma; Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Carcinoma, Endometrioid; Cell Division; Cell Line, Tumor; Cisplatin; Cyclohexanes; Cystadenocarcinoma, Serous; DNA, Neoplasm; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Female; Humans; Mice; Mice, Nude; Neoplasm Proteins; O-(Chloroacetylcarbamoyl)fumagillol; Ovarian Neoplasms; RNA, Neoplasm; Sesquiterpenes; Xenograft Model Antitumor Assays

To investigate the effect of TNP-470 on the proliferation and apoptosis of lung adenocarcinoma cells and to explore the mechanism of the effect.. The TNP-470 with different concentrations was added to cultured lung adenocarcinoma cell strain AGZY-82A and the expressions of proliferating cell nuclear antigen (PCNA), p53, bcl-2, vascular endothelial growth factor (VEGF) and VEGF receptor Flk-1 of the tumor cells were assessed by immunohistochemical methods.. The experimental data showed that TNP-470 inhibit the expressions of VEGF and Flk-1 by tumor cells with dose-dependence. In the group of cells without TNP-470 (control group) the expression could rates of VEGF and Flk-1 were 70.42% and 50%, respectively; while in the group of cells with 10(7) microgram/L TNP-470, the expression rates of VEGF and Flk-1 decreased to 13.2% and 7.2%, respectively. Under the lnfluence of 10(4) microgram/L TNP-470, the expression rates of VEGF, Flk-1 by the tumor cells decreased gradually with prolonging of the action time. After 6 hours of action, the expression rate of VEGF decreased to 14.4% and after 12 hours, the expression rate of Flk-1 decreased to 6.2%. In the control group the proliferation rate of PCNA-marked cells was (87.03 +/- 0.75)% and the positive expression rate of p53-marked cells was (3.67 +/- 0.39)%, while in the 10(7) microgram/L TNP-470 group the proliferation rate and positive rate of p53-marked cells were 0 and (63.28 +/- 0.84)%, respectively. No expression of bcl-2 in both control and low concentration TNP-470 group. Its expression increased gradually as the concentration induced. In the 10(4) microgram/L TNP-470 group the expression of PCNA by tumor cells decreased. But the expression of p53, bcl-2 increased gradually with prolonging of the action time.. TNP-470 could inhibit the proliferation and promote the apoptosis of lung adenocarcinoma cells through reduction of the VEGF autocrine and down-regulation of the VEGF receptor.

Topics: Adenocarcinoma; Antineoplastic Agents; Apoptosis; Cell Division; Cyclohexanes; Down-Regulation; Endothelial Growth Factors; Humans; Lung Neoplasms; Lymphokines; O-(Chloroacetylcarbamoyl)fumagillol; Receptor Protein-Tyrosine Kinases; Receptors, Growth Factor; Receptors, Vascular Endothelial Growth Factor; Sesquiterpenes; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

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

We established peritoneal dissemination-prone subcultures (PCI-43p3) using nude mice by a repetitive in vivo selection of intraperitoneally inoculated PCI-43, a pancreas adenocarcinoma cell line. The subcultures showed upregulated expression of matrix metalloproteinase (MMP)-9, but not MMP-2 in culture supernatants. They also produced increased amounts of vascular endothelial growth factor (VEGF), which was not associated with alterations in isoforms of VEGF mRNA. PCI-43p3 cells attached to cultured mesothelial cell monolayers more readily than did the parent PCI-43 cells. The angiogenesis inhibiting agent, TNP-470, at 30 mg / kg was administered to the model mice, resulting in a prominent suppression of the establishment of peritoneal nodules. The suppression was dependent on the duration of TNP-470 treatment. TNP-470 treatment significantly suppressed proliferation of tumor cells in disseminated nodules, assessed in terms of immunostaining for proliferating cell nuclear antigen (PCNA). TNP-470 did not affect the in vitro attachment between PCI-43p3 and mesothelial cells. The combined data show that anti-angiogenic treatment profoundly suppresses the in vivo process of peritoneal dissemination.

Topics: Adenocarcinoma; Angiogenesis Inhibitors; Animals; Cell Adhesion; Cyclohexanes; Endothelial Growth Factors; Female; Intercellular Adhesion Molecule-1; Lymphokines; Matrix Metalloproteinases; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Neoplasm Proteins; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Pancreatic Neoplasms; Proliferating Cell Nuclear Antigen; Sesquiterpenes; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

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 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 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