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Surgery Today 2009Combination therapy using antiangiogenic and cytotoxic agents is a useful strategy for advanced cancer, but the mechanism has not yet been elucidated. Moreover, there is... (Comparative Study)
Comparative Study
PURPOSE
Combination therapy using antiangiogenic and cytotoxic agents is a useful strategy for advanced cancer, but the mechanism has not yet been elucidated. Moreover, there is a persistent paradox that destroying tumor vasculature with antiangiogenic agents disturbs the delivery of cytotoxic agents. It has been hypothesized that antiangiogenic agents can lead to normalization of tumor vessels that are structurally and functionally abnormal. The normalization means enhancing the deliver of cytotoxic agents. Our purpose was to investigate whether TSU68, a multiple receptor tyrosine kinase inhibitor that targets vascular endothelial growth factor receptor-2 (VEGFR2), platelet-derived growth factor receptor (PDGFR), and fibroblast growth factor receptor (FGFR), would induce the normalization of tumor vessels.
METHODS
TSU68 was administered for 7 days to mice with xenografted tumors. Tumors of interstitial fluid pressure (IFP) were measured before and after administration of agents. Immunofluorescence double staining for CD31 and alpha-SMA was performed, and a medical video endoscopy system with narrowband illumination (NBI) was used to visualize the vascular pattern.
RESULTS
TSU68 treatment decreased IFP significantly. Immunofluorescence double staining showed a significant increase in the fraction of pericyte coverage in the TSU68-treated group. NBI endoscopy showed that many tumor vessels in TSU68-treated mice were pruned and the diameters of remaining vessels were reduced.
CONCLUSION
The data supported our hypothesis of tumor vascular normalization by the antiangiogenic agent TSU68.
Topics: Angiogenesis Inhibitors; Animals; Colon; Colonic Neoplasms; Disease Models, Animal; Drug Delivery Systems; Humans; Indoles; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neovascularization, Pathologic; Neovascularization, Physiologic; Oxindoles; Propionates; Pyrroles; Receptors, Fibroblast Growth Factor; Receptors, Platelet-Derived Growth Factor; Sensitivity and Specificity; Transplantation, Heterologous; Vascular Endothelial Growth Factor Receptor-2; Xenograft Model Antitumor Assays
PubMed: 19997799
DOI: 10.1007/s00595-009-4020-y -
Journal of Molecular Graphics &... Nov 2009Inhibition of tyrosine kinases (such as the epidermal growth factor receptor, EGFR, and/or Abelson leukemia virus protein kinase, ABL) represents a major advancement in...
Inhibition of tyrosine kinases (such as the epidermal growth factor receptor, EGFR, and/or Abelson leukemia virus protein kinase, ABL) represents a major advancement in the treatment of solid tumors, supported by the clinical administration of gefitinib, erlotinib, imatinib, and dasatinib. The identification of the binding interactions in the EGFR/ligands and the ABL/ligands complexes can facilitate the structure-based design of new tyrosine kinase inhibitors. We carried out induced-fit docking studies of 18 structurally diverse kinase inhibitors against the EGFR, the active and inactive states of the ABL protein. Our docking data show that the induced-fit docking (IFD) protocol can successfully reproduce the native poses of ligands from different sources. The binding interactions and the docked poses are consistent with the available experimental data. Our results indicate that imatinib is a weak binder to the active state of ABL but a strong binder to EGFR. The increased sensitivity of erlotinib to EGFR might be attributed to Cys797 of EGFR. In addition to Cys797, other important residues for kinase inhibitor design include Thr790, Met793, Lys745 and Asp855 of EGFR; and Thr315, Met318, Asp381 and Glu286 of the ABL. The minimum number of H-bonds required for the ligand binding provides a reasonable explanation to the effectiveness of nilotinib against most imatinib resistant mutants.
Topics: Binding Sites; Crystallography, X-Ray; Enzyme Activation; ErbB Receptors; Gefitinib; Hydrogen Bonding; Indoles; Ligands; Models, Molecular; Oxindoles; Propionates; Protein Structure, Secondary; Protein-Tyrosine Kinases; Proto-Oncogene Proteins c-abl; Pyrroles; Quinazolines; Reproducibility of Results; Sunitinib
PubMed: 19767223
DOI: 10.1016/j.jmgm.2009.08.012 -
The Journal of Allergy and Clinical... Oct 2009
Thymic stromal lymphopoietin induction by polyinosinic:polycytidylic acid in human keratinocytes is preferentially mediated through protein kinase R and retinoid-inducible gene I and not Toll-like receptor 3.
Topics: 2-Aminopurine; Antimetabolites; Cells, Cultured; Cytokines; DEAD Box Protein 58; DEAD-box RNA Helicases; Enzyme Inhibitors; Humans; Indoles; Interferon Inducers; Interferon-Induced Helicase, IFIH1; Keratinocytes; Macrolides; Oxindoles; Poly I-C; Propionates; Pyrroles; Receptors, Immunologic; Toll-Like Receptor 3; eIF-2 Kinase; Thymic Stromal Lymphopoietin
PubMed: 19767083
DOI: 10.1016/j.jaci.2009.07.028 -
British Journal of Cancer May 2009Dynamic contrast-enhanced (albumin-Gd-DTPA) magnetic resonance imaging, performed during 2 weeks of daily administration of an inhibitor of tyrosine kinase receptors...
Dynamic contrast-enhanced (albumin-Gd-DTPA) magnetic resonance imaging, performed during 2 weeks of daily administration of an inhibitor of tyrosine kinase receptors (SU6668) in an HT-29 colon carcinoma model, revealed the onset of a hyper-enhancing rim, not observed in untreated tumours. To account for tissue heterogeneity in the quantitative analysis, we segmented tumours into three subunits automatically identified by cluster analysis of the enhancement curves using a k-means algorithm. Transendothelial permeability (Kps) and fractional plasma volume (fPV) were calculated in each subunit. An avascular and necrotic region, an intermediate zone and a well-vascularised periphery were reliably identified. During untreated tumour growth, the identified sub-regions did not substantially change their enhancement pattern. Treatment with SU6668 induced major changes at tumour periphery where a significant increase of Kps and fPV was observed with respect to control tumours. Histology revealed a sub-capsular layer composed of hyper-dense viable tumour cells in the periphery of untreated tumours. The rim of viable neoplastic cells was reduced in treated tumours, and replaced by loose connective tissue characterised by numerous vessels, which explains the observed hyper-enhancement. The present data show a peripheral abnormal development of cancer-associated stroma, indicative of an adaptive response to anti-angiogenic treatment.
Topics: Animals; Carcinoma; Cell Proliferation; Colonic Neoplasms; Disease Progression; HT29 Cells; Humans; Indoles; Magnetic Resonance Imaging; Mice; Mice, Nude; Neovascularization, Pathologic; Oxindoles; Propionates; Pyrroles; Receptor Protein-Tyrosine Kinases; Stromal Cells; Up-Regulation; Xenograft Model Antitumor Assays
PubMed: 19384298
DOI: 10.1038/sj.bjc.6605041 -
Drug Metabolism and Pharmacokinetics 2008The anti-angiogenic agent TSU-68 is known to rapidly induce cytochrome P450 activity responsible for its own hydroxylation in rats. In this study, we identified CYP1A1... (Comparative Study)
Comparative Study
The anti-angiogenic agent TSU-68 is known to rapidly induce cytochrome P450 activity responsible for its own hydroxylation in rats. In this study, we identified CYP1A1 and CYP1A2 as the TSU-68-induced P450 and temporally characterized the rapid induction of these isoforms. Protein and mRNA levels of CYP1A1 and CYP1A2 along with CYP1A activities were examined in rat liver after a single oral administration of 500 mg/kg TSU-68. CYP1A-mediated ethoxyresorufin O-deethylation and TSU-68 hydroxylation activities reached the maximum at 12 hr. The activities were maintained up to 24 hr and then slowly decreased down to control levels. Protein levels of both CYP1A1 and CYP1A2 were also rapidly induced with temporal profiles similar to the profile of CYP1A activity. In contrast, unlike CYP1A2 mRNA levels, which peaked at 12 hr and almost returned to control levels by 48 hr, CYP1A1 mRNA levels peaked as early as 3 hr and returned to control levels by 24 hr. Thus, CYP1A1 showed more rapid elevation and turnover of its mRNA than CYP1A2. In conclusion, TSU-68 administered to rats rapidly induced mRNA and protein of CYP1A1 and CYP1A2 as well as CYP1A activity. Furthermore, the data showed a difference in the time-dependent induction between CYP1A1 and CYP1A2 mRNAs.
Topics: Angiogenesis Inhibitors; Animals; Base Sequence; Cytochrome P-450 CYP1A1; Cytochrome P-450 CYP1A2; Enzyme Induction; Gene Expression Regulation, Enzymologic; Indoles; Male; Microsomes, Liver; Molecular Sequence Data; Oxindoles; Propionates; Pyrroles; RNA, Messenger; Rats; Rats, Sprague-Dawley; Time Factors
PubMed: 19122336
DOI: 10.2133/dmpk.23.421 -
Bioorganic & Medicinal Chemistry Letters Feb 2009NP506, the 3-{2,4-dimethyl-5-[2-oxo-5-(N'-phenylhydrazinocarbonyl)-1,2-dihydro-indol-3-ylidenemethyl]-1H-pyrrol-3-yl}-propionic acid, was designed as FGF receptor 1...
NP506, the 3-{2,4-dimethyl-5-[2-oxo-5-(N'-phenylhydrazinocarbonyl)-1,2-dihydro-indol-3-ylidenemethyl]-1H-pyrrol-3-yl}-propionic acid, was designed as FGF receptor 1 inhibitor by computational study and found to be more active against endothelial proliferation of HUVEC after the rhFGF-2 stimulation than SU6668 with minimum effective dose of 10 microM. NP506 inhibited the tyrosine phosphorylation in FGF, VEGF, and PDGF receptors and the activation of extracellular signal-regulated kinase (ERK), c-Jun-N-terminal-kinase (JNK) and AKT after the rhFGF-2 stimulation. The introduction of the phenyl hydrazide motif to the position 5 of the pyrido[2,3-d]pyrimidine scaffold led to the inhibitory effect in two signaling pathways: inhibition of AKT activation in the phosphatidyl inositol 3'-kinase (PI13K)/AKT signaling pathway and the inhibition of ERK and JNK activation in MAPK pathway.
Topics: Cell Proliferation; Cells, Cultured; Chemistry, Pharmaceutical; Drug Design; Endothelium, Vascular; Enzyme Inhibitors; Fibroblast Growth Factor 2; Humans; Indoles; JNK Mitogen-Activated Protein Kinases; Oxindoles; Phosphorylation; Propionates; Proto-Oncogene Proteins c-akt; Pyrimidines; Pyrroles; Receptor Protein-Tyrosine Kinases; Tyrosine
PubMed: 19110422
DOI: 10.1016/j.bmcl.2008.12.023 -
Cancer Research Dec 2008The aim of this study was to investigate the inhibitory effect of TSU68 [(Z)-5-[(1,2-dihydro-2-oxo-3H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-propanoic acid;...
The aim of this study was to investigate the inhibitory effect of TSU68 [(Z)-5-[(1,2-dihydro-2-oxo-3H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-propanoic acid; SU6668], an inhibitor of vascular endothelial growth factor receptor 2, platelet-derived growth factor receptor beta, and fibroblast growth factor receptor 1 (FGFR1), on colon cancer liver metastasis, and to test the hypothesis that TSU68 modulates the microenvironment in the liver before the formation of metastasis. First, we implanted the highly metastatic human colon cancer TK-4 orthotopically into the cecal walls of nude mice, followed by twice-daily administration of TSU68 (400 mg/kg/d) or vehicle. Five weeks of treatment with TSU68 significantly inhibited liver metastasis compared with the control group (P<0.001). Next, we analyzed the gene expression profile in premetastatic liver using microarrays. Microarray and quantitative reverse transcription-PCR analysis showed that mRNA levels for the chemokine CXCL1 were significantly increased in tumor-bearing mice compared with non-tumor-bearing mice. Moreover, CXCL1 expression was significantly decreased by TSU68 treatment. CXCR2 expression was detected predominantly on tumor cells in orthotopic tumors compared with ectopic tumors. The number of migrating neutrophils in premetastatic liver was significantly decreased in the TSU68-treated group (P<0.001). The amount of interleukin-12 (IL-12) p40 in the portal vein was significantly decreased by TSU68 (P=0.02). Blockade of both CXCR2 and IL-12 p40 with a neutralizing antibody significantly inhibited liver metastasis. These results suggest that the CXCL1/CXCR2 axis is important in cancer metastasis and that TSU68 may modulate the premetastatic niche in the target organ through suppression of the inflammatory response, which might be an alternative mechanism used by antiangiogenic agents.
Topics: Animals; Cell Movement; Chemokine CXCL1; Colonic Neoplasms; Gene Expression Profiling; Humans; Indoles; Interleukin-12 Subunit p40; Liver; Liver Neoplasms, Experimental; Male; Mice; Mice, Inbred BALB C; Neutrophils; Oligonucleotide Array Sequence Analysis; Oxindoles; Propionates; Pyrroles; RNA, Messenger; Receptors, Interleukin-8B; Xenograft Model Antitumor Assays
PubMed: 19047154
DOI: 10.1158/0008-5472.CAN-08-1748 -
Zhonghua Wei Chang Wai Ke Za Zhi =... Jul 2008To investigate the effect of Endostatin and SU6668 combined with 5-FU on the growth and metastasis of human colon cancer in vivo and its mechanism.
OBJECTIVE
To investigate the effect of Endostatin and SU6668 combined with 5-FU on the growth and metastasis of human colon cancer in vivo and its mechanism.
METHODS
Metastatic model of human colon cancer was established by orthotopic implantation of human tumor tissue into colon wall of nude mice. Twelve days later, mice were randomly divided into saline water control, Endostatin, SU6668, Endostatin plus SU6668, and Endostatin plus SU6668 and 5-FU groups, intraperitoneal injected respectively every day for four weeks. Six weeks after implication, the tumor weight, inhibition rates, intratumoral microvessel density (MVD) and metastasis were evaluated after the mice were sacrificed.
RESULTS
Compared with the control, tumor growth was significantly inhibited in mice treated respectively with Endostatin, SU6668, Endostatin plus SU6668 and Endostatin plus SU6668 and 5-FU with an inhibition rate of 0, 64.9%, 63.5%, 76.4% and 88.2% respectively,and MVD decreased significantly in treated groups [(18.10+/-5.65) vs (2.75+/-0.75), (3.17+/-0.58), (0.94+/-0.42) and (0.36+/-0.45)]. The incidences of peritoneal and region lymph node metastases were significantly inhibited in Endostatin, SU6668, Endostatin plus SU6668 and Endostatin plus SU6668 and 5-FU (90% vs 16.7%, 25%, 0 and 0; 90% vs 0, 0, 0 and 0). The growth and metastasis of human colon cancer implanted in nude mice were significantly inhibited in Endostatin, SU6668, Endostatin plus SU6668, and Endostatin plus SU6668 and 5-FU, and the effect of Endostatin plus SU6668 and 5-FU was the most obviously.
CONCLUSION
Endostatin plus SU6668 and 5-FU has strong inhibitory effect both on tumor growth and metastasis of human colon cancer.
Topics: Angiogenesis Inhibitors; Animals; Cell Line, Tumor; Colonic Neoplasms; Endostatins; Fluorouracil; Humans; Indoles; Lymphatic Metastasis; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Neovascularization, Pathologic; Oxindoles; Propionates; Pyrroles
PubMed: 18636363
DOI: No ID Found -
Clinical Cancer Research : An Official... Apr 2008Investigations on the combination of radiotherapy with vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) antiangiogenic agents, which...
PURPOSE
Investigations on the combination of radiotherapy with vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) antiangiogenic agents, which has the potential to improve the clinical outcome in cancer patients.
EXPERIMENTAL DESIGN
Here, we analyze the combined VEGF (SU5416) and PDGF (SU6668) receptor tyrosine kinase inhibition with irradiation in human endothelium (HUVEC), prostate cancer (PC3), and glioblastoma (U87) in vitro and in vivo.
RESULTS
Combined inhibition of VEGF and PDGF signaling resulted in enhanced apoptosis, reduced cell proliferation, and clonogenic survival as well as reduced endothelial cell migration and tube formation compared with single pathway inhibition. These effects were further enhanced by additional irradiation. Likewise, in PC3 and U87 tumors growing s.c. on BALB/c nu/nu mice, dual inhibition of VEGF and PDGF signaling significantly increased tumor growth delay versus each monotherapy. Interestingly, radiation at approximately 20% of the dose necessary to induce local tumor control exerts similar tumor growth-inhibitory effects as the antiangiogenic drugs given at their maximum effective dose. Addition of radiotherapy to both mono- as well as dual-antiangiogenic treatment markedly increased tumor growth delay. With respect to tumor angiogenesis, radiation further decreased microvessel density (CD31 count) and tumor cell proliferation (Ki-67 index) in all drug-treated groups. Of note, the slowly growing PC3 tumor responded better to the antiangiogenic drug treatments than the faster-growing U87 tumor. In addition to the beneficial effect of abrogating VEGF survival signaling when combined with radiation, we identified here a novel mechanism for the tumor escape from radiation damage. We found that radiation induced up-regulation of all four isoforms of PDGF (A-D) in endothelial cells supporting adjacent smooth muscle cells resulting in a prosurvival effect of radiation. The addition of SU6668 attenuated this undesirable paracrine radiation effect, which may rationalize the combined application of radiation with PDGF signaling inhibition to increase antitumor effects.
CONCLUSION
A relative low radiation dose markedly enhances local antitumor effects of combined VEGF and PDGF signaling inhibition, suggesting a promising combination regimen for local tumor treatment with radiotherapy remaining an essential element.
Topics: Angiogenesis Inhibitors; Animals; Apoptosis; Cell Line, Tumor; Cell Proliferation; Combined Modality Therapy; Endothelial Cells; Flow Cytometry; Humans; Immunohistochemistry; Indoles; Mice; Mice, Nude; Neoplasms; Neovascularization, Pathologic; Oxindoles; Propionates; Protein Kinase Inhibitors; Pyrroles; Radiation-Sensitizing Agents; Radiotherapy; Receptors, Platelet-Derived Growth Factor; Receptors, Vascular Endothelial Growth Factor; Reverse Transcriptase Polymerase Chain Reaction; Xenograft Model Antitumor Assays
PubMed: 18381963
DOI: 10.1158/1078-0432.CCR-07-1893 -
Drug Metabolism and Disposition: the... Jun 2008(Z)-5-[(1,2-Dihydro-2-oxo-3H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-propanoic acid (TSU-68) is a new anticancer drug that inhibits angiogenic receptor...
Identification of human liver cytochrome P450 isoforms involved in autoinduced metabolism of the antiangiogenic agent (Z)-5-[(1,2-dihydro-2-oxo-3H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-propanoic acid (TSU-68).
(Z)-5-[(1,2-Dihydro-2-oxo-3H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-propanoic acid (TSU-68) is a new anticancer drug that inhibits angiogenic receptor tyrosine kinases, which play a crucial role in tumor-induced vascularization. TSU-68 undergoes hepatic oxidation and glucuronidation. Incubation of TSU-68 with human liver microsomes in the presence of NADPH resulted in the formation of three major metabolites: 5-, 6-, and 7-hydroxyindolinone derivatives. The 5-, 6-, and 7-hydroxylation followed simple Michaelis-Menten kinetics with V(max)/K(m) values (an indicator of intrinsic clearance) of 13, 25, and 6 microl/min/mg, respectively. Of the 10 cDNA-expressed human cytochrome P450 isoforms examined, only CYP1A1 and CYP1A2 exhibited appreciable TSU-68 hydroxylation activity. Inhibition studies with alpha-naphthoflavone (a selective CYP1A2 inhibitor) and anti-CYP1A2 antibody also indicated the almost exclusive role of CYP1A2 in microsomal TSU-68 hydroxylation. Treatment of human hepatocytes with 10 microM TSU-68 resulted in a 28- to 140-fold increase in CYP1A1/2-mediated ethoxyresorufin O-deethylase activity. The protein levels of CYP1A2 were increased in TSU-68-treated hepatocytes, and those of CYP1A1, which were undetectable in control hepatocytes, were also increased to detectable levels in the TSU-68-treated hepatocytes. Thus, TSU-68 was shown to induce CYP1A1/2 expression, which was responsible for its hydroxylation. The observation that TSU-68 treatment resulted in a 10- to 45-fold increase in 5-, 6-, and 7-hydroxylation directly demonstrated the autoinduced hydroxylation of TSU-68. In conclusion, TSU-68 has the potential to cause induction of its own CYP1A1/2-mediated oxidative metabolism in humans. This autoinductive effect provides a clear explanation for the clinically observed decrease in TSU-68 plasma concentrations during repeated administration of the drug.
Topics: Angiogenesis Inhibitors; Cells, Cultured; Cytochrome P-450 Enzyme System; Glucuronides; Hepatocytes; Humans; Hydroxylation; Indoles; Microsomes, Liver; Oxindoles; Propionates; Protein Isoforms; Pyrroles
PubMed: 18322074
DOI: 10.1124/dmd.107.019877