o-(chloroacetylcarbamoyl)fumagillol and Lung-Neoplasms

Lung cancer is a major public health problem and the leading cause of cancer-related death worldwide. Its survival rates have changed little over the past 20 years. The best clinical benefit (ie, survival rates) with combination cytotoxic therapies in non-small-cell lung cancer (NSCLC) may have been reached. The need for improved survival rates in NSCLC has driven the development of novel, rationally designed, targeted therapies. Inhibitors of angiogenesis have been developed and are increasingly studied. Potential targets for therapy include inhibitors of vascular endothelial growth factor receptor, endogenous angiogenesis inhibitors, and cyclooxygenase inhibitors. Combining targeted molecules with traditional cytotoxic therapies usually results in lower required chemotherapy doses and fewer, less severe side effects. A number of ongoing randomized studies are underway to evaluate this idea. It is anticipated that these new targeted therapies will play an important role, along with cytotoxic and radiation therapies, in the management of metastatic disease.

Topics: Angiogenesis Inhibitors; Angiostatins; Carcinoma, Non-Small-Cell Lung; Collagen; Cyclohexanes; Endostatins; Humans; Lung Neoplasms; O-(Chloroacetylcarbamoyl)fumagillol; Peptide Fragments; Plasminogen; Receptor Protein-Tyrosine Kinases; Receptors, Growth Factor; Receptors, Vascular Endothelial Growth Factor; Sesquiterpenes; Survival Rate; Thalidomide

Preclinical studies have demonstrated a synergistic effect with the angiogenesis inhibitor TNP-470 and several cytotoxic agents. A recent clinical trial with the combination of paclitaxel and TNP-470 has shown promising effects. The present study was designed to determine the toxicity and pharmacokinetics of carboplatin in combination with TNP-470 in comparison with the doublet regimen of paclitaxel and carboplatin in patients with solid tumors.. Enrolled in the study were 17 patients with lung (11), head/neck (3), sarcoma (2) and thymoma (1). The patients received intravenous paclitaxel and carboplatin on day 1 followed by TNP-470 (60 mg/m(2) i.v. over 1 h administered thrice weekly on Monday, Wednesday, and Friday). Each cycle of therapy consisted of 3 weeks. The initial cohort of three patients received carboplatin at AUC 5 mg/ml x min. No dose-limiting toxic effects occurred, thus the subsequent cohort received carboplatin at AUC 6 mg/ml x min. In addition to toxicity, the pharmacokinetics of carboplatin were evaluated, and tumor response and patient survival rates were assessed.. The administered regimen of paclitaxel (225 mg/m(2) i.v. over 3 h) and carboplatin (AUC 6 mg/ml x min i.v. over 1 h) on day 1 followed by TNP-470 (60 mg/m(2) i.v. over 1 h administered thrice weekly on Monday, Wednesday, and Friday) was defined as both the maximum tolerated and optimal dose. Hematological toxic effects were similar to those expected with the chemotherapy doublet. All neurocognitive impairments were graded as mild to moderate and reversed after discontinuation of TNP-470 administration. No alterations in the pharmacokinetic disposition of carboplatin were noted. Overall, the median survival duration was 297 days. Four patients (24%) had a partial response, and eight (47%) had stable disease.. The combination of TNP-470, paclitaxel, and carboplatin is a reasonably well tolerated regimen. Further randomized studies of TNP-470 with this doublet regimen are now warranted for non-small-cell lung carcinoma and other solid tumors.

Topics: Adult; Aged; Angiogenesis Inhibitors; Antineoplastic Combined Chemotherapy Protocols; Carboplatin; Cyclohexanes; Female; Humans; Lung Neoplasms; Male; Middle Aged; Neoplasms; O-(Chloroacetylcarbamoyl)fumagillol; Paclitaxel; Sesquiterpenes

Preclinical studies suggested that the antiangiogenic agent TNP-470 was synergistic with cytotoxic therapy. TNP-470 was administered with paclitaxel to adults with solid tumors to define the safety and optimal dose of the combination regimen and to assess pharmacokinetic interactions.. Thirty-two patients were enrolled chronologically onto one of two treatment arms. Arm A involved a fixed TNP-470 dose with escalating doses of paclitaxel, and Arm B involved a fixed paclitaxel dose with escalating doses of TNP-470. Paclitaxel and TNP-470 pharmacokinetics were evaluated along with toxicity.. The combination of TNP-470 administered at 60 mg/m(2) three times per week and paclitaxel 225 mg/m(2) administered over 3 hours every 3 weeks was defined as both the maximum-tolerated dose and the optimal dose. Myelosuppression was similar to that expected with paclitaxel alone. Mild to moderate neurocognitive impairment was observed; however, the majority of changes were subclinical and reversible as determined by prestudy and poststudy neuropsychiatric test results. A clinically insignificant decrease of paclitaxel clearance was observed for the combination. Median survival for all patients was 14.1 months. Partial responses were reported in eight (25%) of 32 patients and in six (38%) of 16 patients with NSCLC, 60% of whom had received prior chemotherapy.. The combination of TNP-470 and paclitaxel, each at full single-agent dose, seems well tolerated, with minimal pharmacokinetic interaction between the two agents. Further studies of TNP-470 with chemotherapy regimens are warranted in NSCLC and other solid tumors.

Topics: Adult; Aged; Angiogenesis Inhibitors; Antibiotics, Antineoplastic; Antineoplastic Agents, Phytogenic; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Non-Small-Cell Lung; Cyclohexanes; Drug Administration Schedule; Female; Humans; Lung Neoplasms; Male; Middle Aged; O-(Chloroacetylcarbamoyl)fumagillol; Paclitaxel; Sesquiterpenes; Treatment Outcome

Cancer-related deaths are caused principally by recurrence and metastasis arising from residual disease, whose therapeutic responses has been suggested to be substantially different from primary tumors. However, experimental animal models designed for evaluating the therapeutic responses of residual disease are mostly lacking. To overcome this deficiency, we have developed a preclinical model that recapitulates the progression for advanced nonsmall cell lung cancer (NSCLC). An archived Lewis lung carcinoma mouse tumor, propagated only through serial in vivo transplantation and never adapted to cell culture, was stably labeled using lentivirus-encoded biomarkers, consistently expressed through an RNA polymerase II promoter. Labeled tumors were inoculated into syngeneic immunocompetent mice to ensure superior tumor-host interactions. Primary tumors were resected on reaching a predetermined size, followed by treatment in a setting akin to postsurgical first-line adjuvant chemotherapy and routine imaging to monitor the progression of pulmonary metastasis. We discovered that efficacious treatment, instead of reducing disease growth rates, significantly prolonged disease-free survival and overall survival. As in the clinic, cisplatin-based regimes were more effective in this model. However, the response of metastases to specific agents could not be predicted from, and often opposed, their effects on subcutaneous "primary" tumors, possibly due to their distinct growth kinetics and host interactions. We here introduce a clinically relevant model of residual metastatic disease that may more accurately predict the therapeutic response of recurrent, metastatic disease.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Lewis Lung; Carcinoma, Non-Small-Cell Lung; Chemotherapy, Adjuvant; Cisplatin; Cyclohexanes; Disease Models, Animal; Disease Progression; Female; Humans; Lung Neoplasms; Mice; Mice, Inbred C57BL; Neoplasm, Residual; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Survival Rate; Treatment Outcome

Targeting angiogenesis, the formation of blood vessels, is an important modality for cancer therapy. TNP-470, a fumagillin analog, is among the most potent and broad-spectrum angiogenesis inhibitors. However, a major clinical limitation is its poor oral availability and short half-life, necessitating frequent, continuous parenteral administration. We have addressed these issues and report an oral formulation of TNP-470, named Lodamin. TNP-470 was conjugated to monomethoxy-polyethylene glycol-polylactic acid to form nanopolymeric micelles. This conjugate can be absorbed by the intestine and selectively accumulates in tumors. Lodamin significantly inhibits tumor growth, without causing neurological impairment in tumor-bearing mice. Using the oral route of administration, it first reaches the liver, making it especially efficient in preventing the development of liver metastasis in mice. We show that Lodamin is an oral nontoxic antiangiogenic drug that can be chronically administered for cancer therapy or metastasis prevention.

Topics: Administration, Oral; Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Cell Line, Tumor; Cyclohexanes; Humans; Lung Neoplasms; Mice; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Polyesters; Sesquiterpenes; Treatment Outcome

Osteopontin (OPN) is a multifunctional cytokine involved in cell signaling by interacting with alphavbeta3 integrins. Recent clinical studies have indicated that OPN expression is associated with tumor progression and poor prognosis among patients with lung cancer. However, the biological role of OPN in human lung cancer has not yet been well-defined. The purpose of this study is to investigate and provide evidence for the causal role of OPN regarding tumor growth and angiogenesis in human lung cancer. In this study, we developed a stable OPN transfectant from human lung cancer cell line SBC-3 which does not express the intrinsic OPN mRNA. To reveal the in vivo effect of OPN on tumor growth of human lung cancer, we subcutaneously injected OPN-overexpressing SBC-3 cells (SBC-3/OPN) and control cells (SBC-3/NEO) into the nude mice. Transfection with the OPN gene significantly increased in vivo tumor growth and neovascularization of SBC-3 cells in mice. These in vivo effects of OPN were markedly suppressed with administration of anti-alphavbeta3 integrin monoclonal antibody or anti-angiogenic agent, TNP-470. Furthermore, recombinant OPN protein enhanced human umbilical vein endothelial cell (HUVEC) proliferation in vitro, and this enhancement was significantly inhibited with the addition of anti-alphavbeta3 integrin antibody. Taken together, these results suggest that OPN plays a crucial role for tumor growth and angiogenesis of human lung cancer cells in vivo by interacting with alphavbeta3 integrin. Targeting the interaction between OPN and alphavbeta3 integrin could be effective for future development of anti-angiogenic therapeutic agents for patients with lung cancer.

Topics: Angiogenesis Inhibitors; Animals; Antibodies, Monoclonal; Cell Proliferation; Cyclohexanes; Female; Humans; Integrin alphaVbeta3; Lung Neoplasms; Mice; Mice, Inbred BALB C; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Osteopontin; Sesquiterpenes; Transfection; Umbilical Cord; Xenograft Model Antitumor Assays

Genetic analysis of a high-metastatic clone of RCT sarcoma (HM-RCT) was the aim of the study. HM-RCT was developed by the lung passage as well as limiting dilution method from the original RCT sarcoma, in which a tumor was spontaneously developed in a C3H/He mouse. HM-RCT expressed enhanced POU domain (class 2, associating factor 1), adenylate cyclase 7, procollagen type III (alpha), A kinase anchor protein 4 and Ehm (expressed on high-metastatic cells) and 11 expressed sequence tags (ESTs). compared with the original clone of RCT. Eighteen specific genes and 14 ESTs were underexpressed in HM-RCT. We investigated the effects of angiogenesis inhibitor TNP-470 on tumor growth and metastasis of this HM-RCT in vivo. In an experimental group, mice received TNP-470 (30 mg/kg) intraperitoneally every other day. After 5 weeks, the growth of the TNP-470-treated tumor was significantly suppressed in vivo, but did not affect the metastasis. The proportion of positive PCNA-stained cells and cellular telomerase activity was significantly low in response to TNP-470.

Topics: A Kinase Anchor Proteins; Adenylyl Cyclases; Angiogenesis Inhibitors; Animals; Collagen Type III; Cyclohexanes; Expressed Sequence Tags; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Lung Neoplasms; Male; Mice; Mice, Inbred C3H; Neoplasm Invasiveness; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Oligonucleotide Array Sequence Analysis; Proliferating Cell Nuclear Antigen; Protein Precursors; Sarcoma, Experimental; Sesquiterpenes; Telomerase; Time Factors; Trans-Activators

Endostatin (ED) is a carboxyl-terminal fragment of type XVIII collagen with a strong anti-angiogenic activity. The purpose of this study is to determine the effect of ED gene transfer into lung cancer cells on in vivo tumor growth in a murine model. The murine lung cancer cell line, Lewis Lung Carcinoma (LLC), was transfected with ED gene to express and secrete ED. After clones were selected to secrete ED, several stable transfectants with ED gene (LLC/ED) and control transfectants (LLC/Mock) were established. In vitro proliferation of these transfectants demonstrated similar growth speed. In contrast to previous reports, in vivo subcutaneous tumorignecity of LCC/ED transfectants was significantly greater than that of LLC/Mock transfectants. Immunohistochemical staining analysis demonstrated that ED gene transfer induced angiogenesis, suggesting coinduction of another gene implicated for neovascularization. As expected, LLC/ED transfectants secreted not only ED but also vascular endothelial growth factor (VEGF) to a much greater degree than LLC/mock transfectants. Interestingly, culture supernatants of LLC/ED cells enhanced in vitro proliferation of human umbilical vein endothelial cells (HUVEC) to a much greater degree than those of LLC/Mock cells. These results indicate that ED gene transfer in murine lung carcinoma cells induces VEGF secretion, resulting in enhancement of in vivo tumorigenecity in the murine model. More attention should be paid for ED gene therapy into lung cancer cells since it may influence other proteins secretion, which upregulates angiogenesis.

Topics: Animals; Cell Line, Tumor; Cyclohexanes; Endostatins; Endothelial Cells; Gene Transfer, Horizontal; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; O-(Chloroacetylcarbamoyl)fumagillol; RNA, Messenger; Sesquiterpenes; Transfection; Up-Regulation; 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

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

Angiogenesis is crucial for tumor growth. Angiogenesis inhibitors, such as O-(chloracetyl-carbamoyl) fumagillol (TNP-470), are thus emerging as a new class of anticancer drugs. In clinical trials, TNP-470 slowed tumor growth in patients with metastatic cancer. However, at higher doses necessary for tumor regression, many patients experienced neurotoxicity. We therefore synthesized and characterized a water-soluble conjugate of N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer, Gly-Phe-Leu-Gly linker and TNP-470. This conjugate accumulated selectively in tumor vessels because of the enhanced permeability and retention (EPR) effect. HPMA copolymer-TNP-470 substantially enhanced and prolonged the activity of TNP-470 in vivo in tumor and hepatectomy models. Polymer conjugation prevented TNP-470 from crossing the blood-brain barrier (BBB) and decreased its accumulation in normal organs, thereby avoiding drug-related toxicities. Treatment with TNP-470 caused weight loss and neurotoxic effects in mice, whereas treatment with the conjugate did not. This new approach for targeting angiogenesis inhibitors specifically to the tumor vasculature may provide a new strategy for the rational design of cancer therapies.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Blood-Brain Barrier; Carcinoma; Chick Embryo; Cyclohexanes; Endothelial Cells; Humans; Liver; Lung Neoplasms; Male; Melanoma; Methacrylates; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, SCID; Molecular Structure; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Polymers; Regeneration; Sesquiterpenes

Early relapse with distant metastasis often is observed in patients with cancer after resection of the primary tumor. It is considered that resection of the primary tumor induces activation of systemic angiogenesis and enhances progression of remote metastasis. The authors show that resection of the primary osteosarcoma tumor enhances progression of pulmonary metastasis in animal osteosarcoma models. Matrigel plug neovascularization assay revealed that systemic angiogenic activity was elevated after primary tumor removal (tumor intact group, 1.61 +/- 0.21 g/dL; tumor removed group, 4.92 +/- 0.35 g/dL). In addition, serum concentration of the angiogenesis inhibitor, endostatin, decreased significantly after primary tumor removal. Treatment with the antiangiogenic reagent TNP-470 suppressed postoperative progression of pulmonary metastasis. These results indicate the possibility that activation of angiogenic activity after resection of osteosarcoma tumors enhances progression of pulmonary metastasis. The current data also suggest that administration of antiangiogenic reagents can prevent progression of pulmonary metastasis in osteosarcoma postoperatively.

Topics: Angiogenesis Inhibitors; Animals; Bone Neoplasms; Collagen; Cyclohexanes; Disease Models, Animal; Endostatins; Endothelial Growth Factors; Female; Intercellular Signaling Peptides and Proteins; Lung Neoplasms; Lymphokines; Mice; Mice, Inbred BALB C; O-(Chloroacetylcarbamoyl)fumagillol; Osteosarcoma; Peptide Fragments; Postoperative Complications; Sesquiterpenes; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

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

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

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

We investigated the effects of the angiogenesis inhibitor TNP-470 on human lung squamous cell carcinoma cell lines H226B and H226Br both in vivo and in vitro. H226B was established from human lung squamous cell carcinoma and H226Br was established from a brain metastatic lesion of H226B in nude mice. Nude mice inoculated with these cells were treated with 30 mg / kg of TNP-470 subcutaneously every other day. At this dose, TNP-470 only significantly suppressed the growth of H226Br tumor, but not H226B tumor. Attempts to use a high dose of TNP-470 (100 mg / kg) resulted in a severe loss of body weight. Immunohistochemical studies showed marked tumor vascularization in H226Br tumor, but the formation of new blood vessels was suppressed by 30 mg / kg of TNP-470. Investigation of the mechanism of anti-angiogenic effects of TNP-470 in vivo showed that the expression and the activity of platelet-derived endothelial cell growth factor / thymidine phosphorylase (PD-ECGF / dThdPase) in H226Br tumor was significantly suppressed by 30 mg / kg of TNP-470. Furthermore, TNP-470 inhibited cell growth of cultured H226Br dose-dependently at concentrations of 1 microg / ml. Immunoblot analysis revealed H226Br cells gave a stronger PD-ECGF signal than H226B cells, and the expression of PD-ECGF / dThdPase in H226Br was also suppressed by treatment with TNP-470 at 0.1 microg / ml. No change in basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF) was noted in these cell lines. Our results suggested that TNP-470 acts, at least in part, by downregulation of PD-ECGF / dThdPase in this cell line.

Topics: Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Body Weight; Carcinoma, Squamous Cell; Cell Division; Cyclohexanes; Down-Regulation; Female; Growth Inhibitors; Humans; Lung Neoplasms; Mice; Mice, Inbred ICR; Mice, Nude; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Thymidine Phosphorylase; Tumor Cells, Cultured

Antiangiogenic therapy is a promising new strategy to inhibit tumor growth and formation of metastases. Vascular endothelial growth factor (VEGF) and its receptors, VEGF-receptor 1 (VEGF-R1; FLT-1) and VEGF-R2 (KDR), have been shown to play a major role in tumor angiogenesis. PTK787/ZK 222584, a specific inhibitor of both VEGF-receptor tyrosine kinases, was investigated for its antitumoral and antiangiogenic activity in a murine renal cell carcinoma model. After intrarenal application of the renal carcinoma cells, mice develop a primary tumor and metastases to the lung and to the abdominal lymph nodes. Daily oral therapy with PTK787/ZK 222584 at a dose of 50 mg/kg resulted in a significant decrease of 61 and 67% in primary tumors after 14 and 21 days, respectively. The occurrence of lung metastases was significantly inhibited at both time points (98% reduction and 78% reduction, respectively). After 14 days, no lymph node metastases developed in the PTK787/ZK 222584-treated group, whereas after 21 days of treatment, the lymph node metastases were reduced by 87%. Vessel density in tumor tissues, detected by immunohistochemistry with an anti-CD31 antibody, was significantly decreased by PTK787/ZK 222584. Using color Doppler imaging ultrasound, significant changes in blood flow in the tumor feeding renal artery were found under treatment with PTK787/ZK 222584. Blood flow changes correlated with changes in vessel density but not with tumor volume. The compound was well tolerated in all in vivo experiments and had no significant effects on body weight or general well-being of the animals. This was in contrast to the animals treated with the antiangiogenic agent TNP-470. s.c. therapy with 30 mg/kg TNP-470 every other day had to be discontinued after 13 days because of animal weight loss (>20%) and ataxia. These results demonstrate that PTK787/ZK 222584 is a potent inhibitor of tumor growth, metastases formation, and tumor vascularization in murine renal cell carcinoma. Furthermore, we have been able to demonstrate that color Doppler imaging ultrasound can be used to measure blood flow to a tumor and that flow correlates with vessel density. Thus, this may be a valuable noninvasive method for monitoring the effects of antiangiogenic agents such as PTK787/ZK 222584 on tumor vasculature.

Topics: Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Carcinoma, Renal Cell; Cell Division; Cyclohexanes; Disease Models, Animal; Enzyme Inhibitors; Female; Kidney Neoplasms; Lung Neoplasms; Lymphatic Metastasis; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Phthalazines; Pyridines; Receptor Protein-Tyrosine Kinases; Receptors, Growth Factor; Receptors, Vascular Endothelial Growth Factor; Renal Circulation; Sesquiterpenes

We examined the effects of tranilast on tumor angiogenesis, tumor growth and metastasis in the mouse Lewis lung carcinoma and C57BL mouse system. Tranilast significantly reduced the dense capillary network induced by Lewis lung cancer cells in a mouse dorsal air sac angiogenesis model. Intraperitoneal administration of tranilast at 200 mg/kg/day reduced the tumor size of mouse Lewis lung carcinoma to about 63% of that of the control and suppressed pulmonary metastasis in a spontaneous system. Immunohistochemistry revealed that tranilast reduced the tumor vascularity and increased apoptosis of the tumor cells in vivo. Tranilast potentiated the inhibition of the tumor growth induced by cyclophosphamide, cis-diamminedichloroplatinum(II), adriamycin and vindesine in vivo. These results suggest that tranilast has antiangiogenic and antitumor effects and might have possible therapeutic applications.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Apoptosis; Carcinoma; Cisplatin; Cyclohexanes; Cyclophosphamide; Doxorubicin; Drug Screening Assays, Antitumor; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; ortho-Aminobenzoates; Sesquiterpenes; Tumor Cells, Cultured; Vindesine

We examined the effects of roxithromycin, a 14-membered ring macrolide antibiotic, on tumor angiogenesis, tumor growth and metastasis of mouse B16BL6 melanoma cells. The inhibitory effect of roxithromycin on angiogenesis using mouse dorsal air sac model was dose-dependent, and 100 mg/kg of roxithromycin administered intraperitoneally twice a day reduced the dense capillary network area to about 20% of the control. Administration of roxithromycin histologically reduced the development of microvessels and mononuclear cell infiltration. In vivo tumor growth studies demonstrated that intraperitoneal administration of roxithromycin at 20 mg/kg/day and 50 mg/kg/day reduced tumor size of B16BL6 melanoma to about 56% and 33% (experiment 1), 71% and 48% (experiment 2) of that in the respective controls. Roxithromycin also significantly inhibited pulmonary metastasis of B16BL6 cells in a spontaneous system. The inhibitory activities of roxithromycin on angiogenesis, tumor growth and metastasis were compared with those of a potent angiogenesis inhibitor, TNP-470. These data demonstrated that roxithromycin has potent antiangiogenic and antitumor effects and might have possible therapeutic applications.

Topics: Air Sacs; Animals; Anti-Bacterial Agents; Antibiotics, Antineoplastic; Cell Movement; Cyclohexanes; Dose-Response Relationship, Drug; Leukocytes, Mononuclear; Lung Neoplasms; Melanoma, Experimental; Mice; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Roxithromycin; Sesquiterpenes; Tumor Cells, Cultured

The effects of the angiogenic inhibitors endostatin, angiostatin, and TNP-470 on tumor growth dynamics are experimentally and theoretically investigated. On the basis of the data, we pose a quantitative theory for tumor growth under angiogenic stimulator/inhibitor control that is both explanatory and clinically implementable. Our analysis offers a ranking of the relative effectiveness of these inhibitors. Additionally, it reveals the existence of an ultimate limitation to tumor size under angiogenic control, where opposing angiogenic stimuli come into dynamic balance, which can be modulated by antiangiogenic therapy. The competitive influences of angiogenically driven growth and inhibition underlying this framework may have ramifications for tissue size regulation in general.

Topics: Angiogenesis Inhibitors; Angiostatins; Animals; Antineoplastic Agents; Collagen; Cyclohexanes; Endostatins; Kinetics; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Models, Biological; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Peptide Fragments; Plasminogen; Sesquiterpenes; Signal Transduction

Topics: Antibiotics, Antineoplastic; Carcinoma, Squamous Cell; Clinical Trials, Phase I as Topic; Cyclohexanes; Female; Humans; Lung Neoplasms; Middle Aged; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Uterine Cervical Neoplasms

The combined effects of TNP-470, a promising antiangiogenic agent, and SN-38, a camptothecin derivative, were evaluated in four human cultured cell lines derived from non-small cell lung cancer (NSCLC). Cytotoxicity experiments were determined by using a tetrazolium salt (MTT) assay. The inhibitory effects of TNP-470 on cell proliferation were dose related and the 50% inhibitory concentrations on these cell lines were 47.3-139.8 microM. Evaluation of drug interactions with isobologram and the combination index values showed that sequential exposure to SN-38 followed by TNP-470 produced synergistic effects in the four cell lines tested. Our findings suggest that such an angiocytotoxic chemotherapy might be promising for the treatment of NSCLC.

Topics: Antineoplastic Combined Chemotherapy Protocols; Camptothecin; Carcinoma, Non-Small-Cell Lung; Cell Division; Cyclohexanes; Drug Screening Assays, Antitumor; Drug Synergism; Humans; Irinotecan; Lung Neoplasms; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Tumor Cells, Cultured

Inhibitory effects of TNP-470, a synthetic analogue of the antibiotic fumagillin secreted by Aspergillus fumigatus, were studied with respect to growth and lung metastasis of the hormone-independent rat prostatic carcinoma cell line AT6.3.. Rat prostatic carcinoma AT6.3 cells were implanted in nude mice subcutaneously. Antimetastatic and growth-inhibitory effects of TNP-470 in vivo were examined 3 weeks after inoculation of AT6.3 cells. Direct growth-inhibitory effect was examined by MTT assay in vitro.. TNP-470 inhibited the growth of AT6.3 cells in vitro. Subcutaneously injected TNP-470 markedly reduced numbers and individual size of lung metastases from AT6.3 cells inoculated percutaneously or intravenously into male BALB/c-nu/nu mice.. This agent, which acts as an angiogenesis inhibitor, may prove to be clinically useful in preventing metastasis of hormone-independent prostatic cancer.

Topics: Animals; Antibiotics, Antineoplastic; Cell Division; Cyclohexanes; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Prostatic Neoplasms; Rats; Sesquiterpenes; Tumor Cells, Cultured

To determine the role of telomerase activity in the growth of tumors in rats undergoing chemotherapy, a comparison of the volumes of telomerase-positive transplantable osteosarcomas was made in rats treated with the antineoplastic agent cis-diammine dichloroplatinum (CDDP) or the angiogenesis inhibitor O-(chloroacetylcarbamoyl)fumagillol (AGM-1470). Male F344 rats, 8 weeks old, received transplants of macroscopic lung metastatic nodules into the subcutaneous back space and treatment was started on day 14 thereafter. CDDP was injected i.v. at doses of 0, 0.625, 1.25 and 2.5 mg/kg body weight (b.w.) and AGM-1470 was administered at total doses of 0, 2.5, 5 and 10 mg/kg b.w. over 2 weeks by osmotic pumps, also implanted into the subcutaneous back space, but remote from the transplanted tumors. On day 28, all animals were killed for measurement of transplanted tumor size and determination of telomerase activities by telomeric repeat amplification protocol (TRAP) assay. The results showed telomerase activity to be highly correlated with the treated/non-treated (T/C) tumor size ratio (r = 0.96, P < 0.0001). In a second experiment, CDDP at 2.5 mg/kg b.w. and AGM-1470 at 10 mg/kg b.w., these being the most effective doses, were given as in the first experiment, and animals were serially killed on days 14, 21, 28, 35 and 42. Tumors in rats treated with CDDP and AGM-1470 showed 18.2% and 20.5% of the control telomerase activity on days 35 and 21, respectively, when tumor growth was inhibited. However, on day 42, the activities increased to 46.5% and 92.5%, this correlating with re-growth (r = 0.73, P < 0.0001). These results suggest that decline of telomerase activity may be involved in tumor growth retardation induced by chemotherapeutic agents. This possibility clearly warrants further mechanistic studies.

Topics: Animals; Antibiotics, Antineoplastic; Biomarkers, Tumor; Bone Neoplasms; Cisplatin; Cyclohexanes; Dose-Response Relationship, Drug; Lung Neoplasms; Male; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Osteosarcoma; Rats; Rats, Inbred F344; Regression Analysis; Sesquiterpenes; Telomerase

The use of surgery for metastatic lung cancer has been established recently and the indications have been extended to multiple and bilateral lung metastases. However, in some patients, secondary lung metastasis appears soon after the first pulmonary surgery, making curative treatment very difficult. Postoperative weakness of tumor angiogenesis suppression mechanisms seems to play an important role in the recurrence of lung metastases. To verify this hypothesis, we performed a clinical and an experimental study.. The clinical study revealed that serum vascular endothelial growth factor (VEGF), also known as vascular permeability factor, increased after pulmonary surgery. The experimental study showed that VEGF played an important role in the rapid growth of dormant micrometastases of the lung. These results suggested that the postoperative increase of VEGF disrupted angiogenesis suppression and induced the growth of dormant micrometastases early in the postoperative period. It was also demonstrated that this effect of VEGF on micrometastases was abolished by AGM-1470, an angiogenesis inhibitor. In conclusion, postoperative treatment with AGM-1470 might inhibit the early recurrence of malignant tumors.

Topics: Adult; Aged; Animals; Antibiotics, Antineoplastic; Cyclohexanes; Endothelial Growth Factors; Female; Humans; Lung Neoplasms; Lymphokines; Male; Mice; Mice, Inbred Strains; Middle Aged; Neoplasm Metastasis; Neoplasm Recurrence, Local; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Pneumonectomy; Postoperative Period; Sesquiterpenes; Survival Rate; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

TNP-740, minocycline, suramin and genistein have demonstrated antiangiogenic activity in various experimental systems. The effect of these agents alone and in two agent combinations on the number of intratumoral vessels and response to cytotoxic anticancer therapies was assessed in animals bearing the Lewis lung carcinoma. Treatment with each of the antiangiogenic agents alone and in two agent combinations decreased the number of intratumoral vessels visualized by CD31 or Factor VIII staining to 30% to 50% of the number in the untreated control tumors. In general, the antiangiogenic agents are more effective adjuvants to cytotoxic therapies when used as two agent combinations than as single agents. The most effective antiangiogenic combinations were: TNP-470/minocycline > TNP-470/genistein > TNP-470/suramin. The increases in the response of the primary tumor to cyclophosphamide, adriamycin, CDDP, BCNU, x-rays or 5-fluorouracil and the lung metastases occur to about the same level with the addition of antiangiogenic agents to the therapies. With the treatment combination TNP-470/minocycline/cyclophosphamide 40% of the animals were cured. The results of these studies indicate that antiangiogenic agents can be very useful additions to treatment regimens for solid tumors.

Topics: Animals; Anti-Bacterial Agents; Antibiotics, Antineoplastic; Antineoplastic Agents, Alkylating; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Lewis Lung; Carmustine; Cell Division; Cisplatin; Cyclohexanes; Cyclophosphamide; Doxorubicin; Drug Synergism; Genistein; Isoflavones; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Minocycline; Neoplasm Invasiveness; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Random Allocation; Sesquiterpenes; Suramin

Topics: Animals; Antibiotics, Antineoplastic; Cyclohexanes; Endothelial Growth Factors; Female; Humans; Lung Neoplasms; Lymphatic Metastasis; Lymphokines; Mice; Mice, Nude; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

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

Antitumor and antimetastatic activity of the angiogenesis inhibitor O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), a semisynthetic analogue of fumagillin, was evaluated in breast cancer cell lines. In an in vitro MTT assay, after 72 hrs continuous exposure to TNP-470, growth inhibition was observed in all seven cell lines of murine (JYG-A, JYG-B, DD-762, and BALB/c-MC) or human (KPL-1, MDA-MB-231, and MKL-F) origin, in which the 50% inhibitory concentrations (IC50) at 72 hrs treatment were 4.6, 4.4, 4.6, 10.1, 35.0, 25.3, and 33.4 micrograms/ml, respectively. In an in vivo assay using JYG-A, JYG-B, KPL-1, and MDA-MB-231 cells by orthotopic (right thoracic mammary fat pad) transplantation in female nude mice, TNP-470 at 30 or 50 mg/kg body weight was injected s.c. every other day from the day of tumor cell inoculation until the end of the experiment. The inhibitory effect on primary tumor growth was obtained in all four cell lines in a dose-dependent manner. In the 50 mg/kg TNP-470-treated group, the reductions in tumor weight of the JYG-A, JYG-B, KPL-1, and MDA-MB-231 cells with respect to the controls were 50%, 30%, 4%, and 49%, respectively. Metastasis was seen in the JYG-A, JYG-B, and KPL-1 cells. The numbers of mice bearing pulmonary metastases of JYG-A and JYG-B cells and regional axillary lymph node metastases of KPL-1 cells were reduced, and TNP-470 at the 50 mg/kg dose to KPL-1 cells significantly reduced lymph node metastases compared with the control. Although the weight gain was retarded in the TNP-470-treated mice, weight loss was not seen. TNP-470 was highly effective in the treatment of breast cancer cells. These results suggest that the clinical use of TNP-470 may be a promising treatment for breast cancer patients.

Topics: Animals; Antibiotics, Antineoplastic; Breast Neoplasms; Cell Division; Cyclohexanes; Female; Humans; Lung Neoplasms; Lymphatic Metastasis; Mammary Neoplasms, Animal; Mammary Neoplasms, Experimental; Mice; Mice, Nude; Neoplasm Metastasis; Neoplasm Transplantation; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Tumor Cells, Cultured

The efficacy of combination therapy with cis-diammine-dichloroplatinum (II) (CDDP) and o-(chloroacetyl-carbamoyl) fumagillol (AGM-1470) was evaluated experimentally using a transplantable rat osteosarcoma line, previously established in our laboratory, with a high potential for metastasis. Tumor-bearing male Fischer 344 rats were administered CDDP (2.5 mg/kg) together with, or after discontinuation of, AGM-1470 treatment (10 mg/kg/body weight/week). When CDDP was administered three days after discontinuation of AGM-1470 the most pronounced antimetastatic effects were observed, although the antitumor effect was approximately the same.

Topics: Animals; Antibiotics, Antineoplastic; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Bone Neoplasms; Cisplatin; Cyclohexanes; Dermatologic Surgical Procedures; Drug Administration Schedule; Injections, Intravenous; Injections, Subcutaneous; Lung Neoplasms; Male; Neoplasm Transplantation; O-(Chloroacetylcarbamoyl)fumagillol; Osteosarcoma; Rats; Rats, Inbred F344; Remission Induction; Sesquiterpenes; Skin Neoplasms; Time Factors

The plasminogen activator system has been found to modulate neoplastic spread and angiogenesis in tumors. An angiogenesis inhibitor, TNP-470, has been shown to possess potent antitumor activities in various types of cancer cells. We therefore investigated the inhibitory effect of TNP-470 on cancer cell proliferation, and the suppressive effect of TNP-470 on urokinase-type plasminogen activator (u-PA) and its inhibitor (PAI-1), in human lung cancer cell lines in vitro. TNP-470 inhibited cell proliferation in a dose-dependent manner in both cell lines. u-PA and PAI-1 in culture supernatants were suppressed with the concentrations of TNP-470, in association with a decrease in viable cancer cells. Unchanged serum levels of u-PA and PAI-1 would be of great advantage to the diseased patients, since the plasminogen activator system has a crucial function in the process of distant metastasis.

Topics: Antibiotics, Antineoplastic; Blotting, Western; Cell Division; Cell Survival; Cyclohexanes; Humans; Lung Neoplasms; O-(Chloroacetylcarbamoyl)fumagillol; Plasminogen Activator Inhibitor 1; Plasminogen Activators; Sesquiterpenes; Tumor Cells, Cultured; Urokinase-Type Plasminogen Activator

We treated a murine osteosarcoma cell line, LM8, which preferentially metastasizes to the lungs, with a new angiogenesis inhibitor, TNP-470, to evaluate the efficacy of this compound in the suppression of pulmonary metastasis of osteosarcoma. In an in vivo experiment, tumor cells were inoculated i.v. into C3H mice, and TNP-470 or vehicle alone (control group) was administered s.c. every day for 3 weeks. In the TNP-470-treated groups, both the number of pulmonary metastatic nodules and the lung wet weight were significantly reduced in a dose-dependent manner. Similarly, vascular density in the metastatic tumors estimated by immunohistochemical staining with anti-von-Willebrand factor antibody as an endothelial marker were significantly reduced. No severe side-effects were found. In an in vitro experiment, viable tumor cells were counted after 3 days' treatment with TNP-470. The 50% inhibitory concentration was 0.6 ng/ml for LM8, which was more sensitive than other tumor cells previously reported. Our results show that TNP-470 suppresses the pulmonary metastasis of LM8 and suggest that both its anti-angiogenic activity and cytostatic activity towards LM8 are responsible for the antitumor effect.

Topics: Animals; Antibiotics, Antineoplastic; Bone Neoplasms; Cell Division; Cyclohexanes; Lung Neoplasms; Male; Mice; Mice, Inbred C3H; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Osteosarcoma; Sesquiterpenes; Tumor Cells, Cultured

In cancer patients, dormant micrometastases are often asymptomatic and clinically undetectable, for months or years, until relapse. We have studied dormant lung metastases under angiogenesis suppression in mice. The metastases exhibited rapid growth when the inhibition of angiogenesis was removed. Tumour cell proliferation, as measured by bromodeoxyuridine incorporation and immunohistochemical staining proliferating cell nuclear antigen, was not significantly different in dormant and growing metastases. However, tumour cells of dormant metastases exhibited a more than threefold higher incidence of apoptosis. These data show that metastases remain dormant when tumour cell proliferation is balanced by an equivalent rate of cell death and suggest that angiogenesis inhibitors control metastatic growth by indirectly increasing apoptosis in tumour cells.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; Carcinoma, Lewis Lung; Cell Division; Cyclohexanes; Fibrosarcoma; Lung; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Organ Size; Proliferating Cell Nuclear Antigen; S Phase; Sesquiterpenes; Tumor Cells, Cultured

The efficacy of the anti-angiogenic agent, O-(chloroacetyl-carbamoyl)fumagillol (AGM-1470), against primary tumor growth and spontaneous lung metastasis was evaluated experimentally using a transplantable osteosarcoma line in rats previously established in our laboratory. Male Fischer 344 rats bearing the tumor with a high potential for metastasis received intermittent or continuous subcutaneous administrations of AGM-1470. Both treatment regimens resulted in significant inhibitions of spontaneous lung metastasis and primary tumor growth in a dose-dependent manner, with continuous administration of AGM-1470 exerting the most pronounced inhibitory effects on both parameters.

Topics: Animals; Antibiotics, Antineoplastic; Cyclohexanes; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Injections, Subcutaneous; Lung Neoplasms; Male; O-(Chloroacetylcarbamoyl)fumagillol; Osteosarcoma; Rats; Rats, Inbred F344; Sesquiterpenes; Tumor Cells, Cultured

The antitumor effect of the novel angiogenesis inhibitor O-(Chloroacetyl-carbamoyl) fumagillol, TNP-470 (TNP, s.c.), a synthetic analogue of fumagillin, was studied in combination with cytotoxic agents--mitomycin C (MMC, i.p.), Adriamycin (i.p.), cisplatin (i.p.), and 5-fluorouracil (i.p.), using B16BL6 melanoma (B16 M) and Lewis lung carcinoma in C57BL/6 mice. When the mice were treated on days 3 and 5, addition of MMC (total dose, 5 mg/kg) or 5-fluorouracil (140 mg/kg) to TNP (150 mg/kg) maximally reduced s.c. B16 M volume from 60 to 15% or from 68 to 40% of control, respectively, and addition of MMC (5 mg/kg) to TNP (150 mg/kg) reduced s.c. Lewis lung carcinoma volume from 75 to 62% of control (P < 0.02, compared to the corresponding single drug treatments). During treatment on days 3, 5, 7, 9, and 11, addition of MMC (5 mg/kg) to TNP (150 mg/kg) reduced s.c. B16 M volume from 43 to 6% of control and reduced the number of pulmonary metastasis of i.v. B16 M from 26 to 5% of control (P < 0.001). For established tumors (> 5 mm in maximal diameter), addition of MMC (12-14 mg/kg), Adriamycin (15-17.5 mg/kg), or cisplatin (4 mg/kg, by one shot) to TNP (120-140 mg/kg) with a 6-7 fractionated dosing schedule reduced s.c. B16 M volume from 50 to 20, 24, or 31% of control and reduced s.c. Lewis lung carcinoma volume from 52 to 34, 27, or 34% of control, respectively (P < 0.02). The effect of combination therapy was additive and dose-dependent, and the earlier fractionated dosing schedule exerted more enhanced antitumor effects. TNP reduced the body weight by approximately 10% of control at maximum, but this toxicity was reversible and was not affected by addition of the cytotoxic agents. The results suggest that the combination of angiogenesis inhibitor TNP and standard cytotoxic agents can be a beneficial addition to the treatment of solid tumors.

Topics: Animals; Antibiotics, Antineoplastic; Antineoplastic Combined Chemotherapy Protocols; Body Weight; Cyclohexanes; Doxorubicin; Drug Synergism; Female; Fluorouracil; Lung Neoplasms; Mice; Mice, Inbred C57BL; Mitomycin; Neoplasms, Experimental; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes

Neovascularization is a critical component for the growth of tumors and is a dominant feature in diseases such as diabetic retinopathy and hemangiomas in infancy. Angiogenesis inhibition is a potentially important therapeutic modality. We have previously reported that AGM-1470 is a fungal-derived angiogenesis inhibitor that suppresses primary tumor growth and metastases and is also nontoxic. alpha-Interferon, an angiogenesis inhibitor, is effective in the treatment of life-threatening hemangiomas. We therefore attempted to treat murine primary tumors and metastases with a combination of AGM-1470 and alpha/beta-interferon. Treatment began after solid tumors formed. Six-week-old syngeneic C57BI/6 mice were treated for 21 days with either AGM-1470, or alpha/beta-interferon or AGM-1470 + alpha/beta-interferon. The combination of the angiogenesis inhibitors AGM-1470 and alpha/beta-interferon suppressed tumor growth by 80% compared with controls (P < or = .001). AGM-1470 and alpha/beta-interferon inhibited pulmonary metastatic tumor growth greater than sevenfold (P < or = .001) compared with controls. These effects were better than either inhibitor alone, and the combined effect was additive. Combination of angiogenesis inhibitors may be useful in the treatment of tumors and other angiogenesis-dependent diseases.

Topics: Animals; Antibiotics, Antineoplastic; Antineoplastic Combined Chemotherapy Protocols; Carcinoma; Cyclohexanes; Drug Screening Assays, Antitumor; Drug Synergism; Interferon-alpha; Interferon-beta; Lung; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Time Factors

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

Antitumor activity of angiogenesis inhibitor TNP-470 was evaluated in eight human cultured cell lines derived from choriocarcinoma: SCH, NUC-1, and GCH-1(m); ovarian cancer; TYK and Nakajima; and uterine endometrial cancer: HEC-6, HEC-50, and HEC-1-A. After 7-day culture with TNP-470, in medium at the concentration of 10(1) to 10(-2) micrograms/ml, the inhibition of growth was observed in all of the eight cell lines. The 50% inhibitory concentration of choriocarcinona cell lines was at an extremely low level compared to that of epithelial ovarian cancer and uterine endometrial cancer. In addition, the antitumor effect of this compound was studied in in vivo experiments using nude mice with tumors of GCH-1(m), NUC-1, or Nakajima cells. When the size of the transplanted tumor reached 100-200 mm3 in volume, 3, 10, or 30 mg/kg of TNP-470 was injected s.c. every other day. The inhibitory effect of TNP-470 was obtained by the administration of 10 and 30 mg/kg in GCH-1(m) and NUC-1 cells, respectively, while in Nakajima cells no significant effect was observed. In nude mice treated with 30 mg/kg of TNP-470, lung metastasis of GCH-1(m) cells was strongly inhibited both in the number and in the size of tumor nodules, indicating that the capillary growth in the originally developed tumor was also significantly reduced. These results suggest that the clinical setting using TNP-470 may be one of the promising treatments for the metastasis of tumor cells.

Topics: Animals; Cell Division; Choriocarcinoma; Cyclohexanes; Drug Screening Assays, Antitumor; Endometrial Neoplasms; Female; Humans; In Vitro Techniques; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Metastasis; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Ovarian Neoplasms; Sesquiterpenes; Tumor Cells, Cultured

The synthetic angiogenesis modulator-1470 O-(chloroacetyl-carbamoyl, AGM-1470) is a potent inhibitor of neovascularization. We have investigated the potential influence of this inhibitor in tumor immunobiology using both in vivo and in vitro models. Mice given a single tail-vein injection of tumor cells were later treated wtih AGM-1470 by s.c. injection. After tumor injection, the lungs were evaluated for macroscopic tumor nodules. AGM-1470 significantly reduced the development of macroscopic pulmonary disease but did not eliminate disease. However, tumor-bearing mice treated with AGM-1470 had significantly reduced spleen weight compared to controls. To determine if the observed decrease in spleen weight in the treated animals was associated with immunosuppression, we studied the possible immunomodulatory effects of AGM-1470. AGM-1470 induced no changes in spleen-cell viability compared to controls. However, addition of angioinhibin at the beginning of IL-2-induced spleen-cell activation significantly inhibited the development of NK-mediated tumor-cell killing. Similarly, splenic T-cell proliferation induced by a mitogenic monoclonal antibody to murine T cells was significantly inhibited when activated in the presence of AGM-1470. The in vitro studies were extended by evaluation of immune system status in tumor-bearing mice treated with AGM-1470. In vivo therapy with AGM-1470 did not significantly change the mean splenic lymphocyte counts and CD4/CD8 ratios from control values. In addition, the induction of splenic NK-mediated tumor killing with IL-2 as well as mitogen-induced T-cell activation was not significantly different from control values. These results suggest that AGM-1470 inhibits tumor growth by blocking neovascularization and may, under certain conditions of drug administration, inhibit immune system function.

Topics: Animals; Antibiotics, Antineoplastic; Cyclohexanes; Female; Immunity; Killer Cells, Natural; Lung Neoplasms; Mice; Mice, Inbred C57BL; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Signal Transduction; Spleen; T-Lymphocytes