o-(chloroacetylcarbamoyl)fumagillol and Brain-Neoplasms

Topics: Animals; Antineoplastic Agents; Brain Neoplasms; Cyclohexanes; Endothelial Growth Factors; Epidermal Growth Factor; Fibroblast Growth Factor 1; Fibroblast Growth Factor 2; Glioma; Humans; Lymphokines; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Prognosis; Sesquiterpenes; Suramin; Transforming Growth Factor alpha; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

This study investigated the effect of TNP-470 in combination with carmustine (BCNU) on the growth of subcutaneously implanted human glioblastoma xenografts in nude mice. Human glioblastoma U-251 cells (1×10(7)) were injected into 24 nude mice subcutaneously. The tumor-bearing mice were randomly divided into 4 groups on the seventh day following tumor implantation: TNP-470 group, in which TNP-470 was given 30 mg/kg subcutaneously every other day 7 times; BCNU group, in which 20 mg/kg BCNU were injected into peritoneal cavity per 4 days 3 times; TNP-470 plus BCNU group, in which TNP-470 and BCNU were coadministered in the same manner as in the TNP-470 group and the BCNU group; control group, in which the mice were given 0.2 mL of the mixture including 3% ethanol, 5% acacia and 0.9% saline subcutaneously every other day 7 times. The tumor size and weights were measured. The tumor microvessel density (MVD) was determined by immunostaining by using goat-anti-mouse polyclonal antibody CD105. The results showed that on the 21th day following treatment, the volume of xenografts in the TNP-470 plus BCNU group was (108.93±17.63)mm(3), markedly lower than that in the TNP-470 group [(576.10±114.29)mm(3)] and the BCNU group [(473.01±48.04)mm(3)] (both P<0.01). And the xenograft volume in these 3 treatment groups was even much lower than that in the control group [(1512.61±470.25) mm(3)] (all P<0.01). There was no significant difference in the volume of xenografts between the TNP-470 group and the BCNU group (P>0.05). The inhibition rate of the tumor growth in the TNP-470 plus BCNU group was (92.80±11.37)%, notably higher than that in the TNP-470 group [(61.91±6.29)%] and the BCNU group [(68.73±9.65)%] (both P<0.01) on the 21th day following treatment. There was no significant difference in the inhibition rate of tumor growth between the TNP-470 group and the BCNU group (P>0.05). The MVD of xenografts in the TNP-470 plus BCNU group was decreased significantly as compared with that in the TNP-470 group or the BCNU group (both P<0.05). The MVD of xenografts in the 3 treatment groups was markedly reduced as compared with that in the control group (all P<0.05). No significant changes in weights were observed before and after the treatment in each group (all P>0.05). It was concluded that the combination of TNP-470 and BCNU can significantly inhibit the growth of human glioblastoma xenografts in nude mice without evident side effects.

Topics: Angiogenesis Inhibitors; Animals; Antibiotics, Antineoplastic; Antineoplastic Agents, Alkylating; Antineoplastic Combined Chemotherapy Protocols; Brain Neoplasms; Carmustine; Cell Line, Tumor; Cyclohexanes; Female; Glioblastoma; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Xenograft Model Antitumor Assays

In an era when molecular and targeted anticancer therapeutics is a major focus and when understanding drug dynamics in tumor is critical, it seems advantageous to be able to relate drug concentrations in tumors to corresponding biological end points. To that end, a novel method, based on physiologically based hybrid pharmacokinetic models, is presented to predict human tumor drug concentrations. Such models consist of a forcing function, describing the plasma drug concentration-time profile, which is linked to a model describing drug disposition in tumors. The hybrid models are originally derived from preclinical data and then scaled to humans. Integral to the scale-up procedure is the ability to derive human forcing functions directly from clinical pharmacokinetic data. Three examples of this approach are presented based on preclinical investigations with carboplatin, topotecan, and temozolomide. Translation of these preclinical hybrid models to humans used a Monte Carlo simulation technique that accounted for intrasubject and intersubject variability. Different pharmacokinetic end points, such as the AUC tumor, were extracted from the simulated human tumor drug concentrations to show how the predicted drug concentrations might be used to select drug-dosing regimens. It is believed that this modeling strategy can be used as an aid in the drug development process by providing key insights into drug disposition in tumors and by offering a foundation to optimize drug regimen design.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Brain Neoplasms; Carboplatin; Computer Simulation; Cyclohexanes; Dacarbazine; Female; Humans; Male; Models, Biological; O-(Chloroacetylcarbamoyl)fumagillol; Ovarian Neoplasms; Predictive Value of Tests; Rats; Rats, Inbred F344; Rats, Nude; Sesquiterpenes; Temozolomide; Topotecan; Transplantation, Heterologous; Vascular Endothelial Growth Factor A

Inhibition of angiogenesis has been shown to reduce tumor growth, metastasis, and tumor microvascular density in experimental models. To these effects we would now like to add induction of differentiation, based on biological analysis of xenografted human neuroblastoma (SH-SY5Y, WAG rnu/rnu) treated with the angiogenesis inhibitor TNP-470. Treatment with TNP-470 (10 mg/kg s.c., n = 15) reduced the tumor growth by 66% and stereological vascular parameters (Lv, Vv, Sv) by 36-45%. The tumor cell apoptotic fraction increased more than threefold, resulting in a decrease in viable tumor cells by 33%. In contrast, the mean vascular diameter (29 microm) and the mean tumor cell proliferative index (49%) were unaffected. TNP-470-treated tumors exhibited striking chromaffin differentiation of neuroblastoma cells, observed as increased expression of insulin-like growth factor II gene (+88%), tyrosine hydroxylase (+96%), chromogranin A, and cellular processes. Statistical analysis revealed an inverse correlation between differentiation and angiogenesis. It is suggested that by inhibiting angiogenesis, TNP-470 induces metabolic stress, resulting in chromaffin differentiation and apoptosis in neuroblastoma. Such agonal differentiation may be the link between angiostatic therapy and tumor cell apoptosis.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; Brain Neoplasms; Cell Differentiation; Chromaffin Cells; Chromogranin A; Chromogranins; Cyclohexanes; Female; Humans; Insulin-Like Growth Factor II; Male; Neoplasm Transplantation; Neovascularization, Pathologic; Neuroblastoma; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Rats, Nude; Sesquiterpenes; Tumor Cells, Cultured; Tyrosine 3-Monooxygenase

Hemangiomas represent the most frequent tumors of infancy. However, the pathogenesis of these tumors is still largely unknown, and current treatment of juvenile hemangiomas remains unsatisfactory. Here we present a novel animal model to study proliferating hemangiomas and to evaluate the effect of angiostatic compounds on their growth. Intraperitoneal (i.p.) infection of 4-day-old rats with murine polyomavirus resulted in the development of multiple cutaneous, intramuscular (i.m.), and cerebral hemangiomas with 100% frequency. Histological examination of the brain revealed the formation of immature lesions as soon as 4 days postinfection (p.i.). The subsequent exponential growth of the hemangiomas, both in number and size, was associated with severe hemorrhage and anemia. The cerebral, cutaneous, and i.m. lesions consisted of blood-filled cysts, histologically similar to human cavernous hemangiomas and stained positive for proliferating cell nuclear antigen, urokinase-type plasminogen activator, and vascular endothelial growth factor. Mature cerebral hemangiomas also expressed von Willebrand factor. Cerebral lesions caused death of the untreated animals within 19.2 +/- 1.1 days p.i. Remarkably fewer and smaller hemangiomas developed in animals that had been treated s.c. with the angiogenesis inhibitor TNP-470. Accordingly, TNP-470 (50 mg/kg), administered twice a week from 3 days p.i., significantly delayed tumor-associated mortality [mean day of death, 28.2 +/- 3.3 (P < 0.001)]. Even if therapy was initiated when cerebral hemangiomas were already macroscopically visible (i.e., 9 days p.i.), a significant delay in hemangioma-associated mortality was observed. Also, the IFN-inducer polyinosinic-polycytidylic acid caused a delay of 9 days (P < 0.005) in tumor-associated mortality when administered i.p. at 5 mg/kg, twice a week, starting at day 3 p.i. The model described here may be useful for investigating (a) the angiogenic mechanism(s) underlying hemangioma progression; and (b) the effect of anti-angiogenic compounds on vascular tumor growth.

Topics: Animals; Animals, Newborn; Antibiotics, Antineoplastic; Body Weight; Brain Neoplasms; Cell Transformation, Viral; Cyclohexanes; Disease Models, Animal; Disease Progression; Endothelium, Vascular; Female; Hemangioma; Humans; Infant; Infant, Newborn; Injections, Subcutaneous; Interferon Inducers; Male; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Poly I-C; Polyomavirus; Polyomavirus Infections; Rats; Sesquiterpenes; Sex Factors; Tumor Virus Infections; Viral Load

The angiogenic phenotype is associated with hyperpermeable capillaries. Through treatment with angiogenesis inhibitors capillary permeability may be reduced, and it can be anticipated that cytotoxic agents coadministered may be adversely affected. The current investigation examined this possibility for the combination of TNP-470, an angiogenesis inhibitor, and temozolomide (TMZ), a DNA-alkylating agent with demonstrated activity in brain tumors. TNP-470 (30 mg/kg) was given s.c. on days 6, 8, 10, 12, and 14 following s.c. implantation of rat C6 glioma cells in Sprague-Dawley rats. On the 15th day following tumor implantation, control (no TNP-470) and treated rats received 40 mg/kg of TMZ intraarterially. Prior to dosing, a linear microdialysis probe was placed in the tumor to collect interstitial fluid. Plasma and interstitial fluid samples were collected for 8 h and measured for TMZ by a high-performance liquid chromatography assay. Pharmacokinetic parameters for TMZ were calculated by noncompartmental methods. Total systemic clearance (39.8 +/- 7 versus 44.2 +/- 14 ml min(-1) kg(-1)) and volume of distribution (5.4 +/- 2 versus 5.2 +/- 0.8 L kg(-1)) were not significantly different in control and TNP-470-treated animals. However, the mean TMZ area under the interstitial fluid concentration-time curve was reduced by 25% in the TNP-470-treated group compared to the control (5450 +/- 1892 versus 4120 +/- 1790 micrograms min ml(-1); P < 0.05). It appears that TNP-470 caused this reduction in the tumor uptake of TMZ by its pharmacodynamic action on the tumor vasculature. Since combination regimens using angiogenesis inhibitors and cytotoxic drugs will be needed to determine how such combinations can be used effectively. The current animal model, which utilized tumor microdialysis, can serve as a model to further analyze combination chemotherapy.

Topics: Animals; Antibiotics, Antineoplastic; Antineoplastic Agents, Alkylating; Brain Neoplasms; Cyclohexanes; Dacarbazine; Factor VIII; Glioma; Microscopy, Confocal; Neoplasm Transplantation; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Sesquiterpenes; Temozolomide; Tumor Cells, Cultured

A novel angiogenesis inhibitor TNP-470 was investigated for its anti-tumor activity against malignant gliomas both in vitro and in vivo. TNP-470 cytostatically inhibited the growth in all of the seven glioma cell lines in culture including anticancer drug resistant cells. The 50% inhibitory concentrations (IC50) of these glioma cell lines were 10 to 30 micrograms/ml and they were 10 to 20 times higher than IC50 of normal endothelial cells. TNP-470 (30 mg/kg, i.p., every other day) also significantly inhibited the tumor growth of T98G-transplanted nude mice. Microscopically, tumor vessels after the treatment of the tumor-bearing mice with TNP-470 became fewer in number and smaller in diameter than those without treatment. Furthermore, there appeared extensive necrotic areas in the tumor with TNP-470. These results indicate that TNP-470 is a potent angiogenesis inhibitor for malignant gliomas. In addition, the studies of labeling index of BrdU and Ki67 suggest that TNP-470 may act mainly on tumor endothelial cells, thus resulting in reduction of the tumor growth.

Topics: Animals; Antibiotics, Antineoplastic; Brain Neoplasms; Cyclohexanes; Drug Resistance; Drug Screening Assays, Antitumor; Glioma; Humans; Mice; Mice, Nude; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Sesquiterpenes; Tumor Cells, Cultured

Angiogenesis is a process fundamental to the growth of many solid tumours. Agents that inhibit angiogenesis may have a role in preventing tumour growth. AGM-1470, a synthetic analogue of fumagillin, has been shown to inhibit tumour growth in several extracranial solid tumour models. Its use has been reported to have minimal side effects. No studies have been reported using AGM-1470 in the treatment of an intracranial tumour. To determine the effect of AGM-1470 on an intracranial glial tumour, we used an improved implantation technique to place 80,000 9L tumour cells into the right caudate nucleus of 25 male Fischer 344 rats. Starting on the first post-implantation day, 12 animals received 30 mg kg-1 AGM-1470 via intraperitoneal injection every other day until death. Thirteen control animals received vehicle only. Evidence of intracranial tumour was apparent in 22/23 animals (96%). All animals treated with AGM-1470 experienced a progressive and significant weight loss when compared to controls. At day 17, treated animals retained 80.0 +/- 2.2% of their initial weight, (mean +/- SD) compared to 100.9 +/- 3.6% for controls (p = 2.25 x 10(-12); student's t-test). AGM-1470 had no effect on survival. Median survival in the treatment group was 24.5 days compared to 25 days in the controls (p = 0.95; Mann-Whitney). AGM-1470, although promising in extracranial tumour models, may not be as effective in controlling the growth of intracranial tumours, and its use is not without significant systemic effects. More studies are needed before this drug is used in human brain tumour trials.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Antibiotics, Antineoplastic; Brain Neoplasms; Cyclohexanes; Disease Models, Animal; Gliosarcoma; Male; Neoplasm Transplantation; O-(Chloroacetylcarbamoyl)fumagillol; Rats; Rats, Inbred F344; Sesquiterpenes; Tumor Cells, Cultured

Glioblastoma is the most malignant primary brain tumor. Inhibition of angiogenesis is one potential strategy for treating this fatal hypervascular tumor. AGM-1470 (also called TNP-470), a novel, potent, fungus-derived inhibitor of angiogenesis, was tested on the growth of human glioblastoma cells in culture and on the growth of the tumor in nude mice. In nude mice with subrenally implanted U-87 MG glioblastomas, AGM-1470 significantly inhibited tumor growth (P < 0.01), and in nude mice with intracranial U-87 MG glioblastomas, AGM-1470 prolonged survival. In addition to its expected action as an angiogenesis inhibitor, AGM-1470 also directly inhibited U-87 MG cells in culture at concentrations similar to those that inhibited endothelial cells. The combined inhibition of glioblastoma cell mitosis and of glioblastoma-induced neovascularization suggests that AGM-1470 should be considered for further investigation in the treatment of this fatal tumor.

Topics: Animals; Biomarkers, Tumor; Brain Neoplasms; Cell Division; Cell Line; Cyclohexanes; Dose-Response Relationship, Drug; Endothelium, Vascular; Female; Glioblastoma; Humans; Mice; Mice, Nude; Neoplasm Transplantation; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Subrenal Capsule Assay; Tumor Cells, Cultured; von Willebrand Factor