Anti-tumor Activity Of New Sulfur-Nitrogen Heterocyclic Compound In Non-small Cell Lung Cancer Models

Cancer as a kind of diseases influence of all ages, along with the increase of age the risk of cancer also increases. According to the statistics, there were7.9millions people dead of cancer in2007, accounted for13%death coursed by diseases in worldwide. With the arrivaling of the aging society and the worsening of the living environment, the occurrence of cancer is also constant rise1. The most common cancer that causes death is lung cancer, about1.3million people dead of lung cancer in2004, reported by the world health organization. All the lung cancer patients about80%belong to non-small cell lung cancer; in contrast, only20%are small lung cancer. There are many factors affecting the treatment of lung cancer, as different lung cancer stage it is should be choose different treatment, and chemotherapy treatment is widely used. At present, the drug resistance and the toxicity of chemotherapy drugs are the most important challage should be facing. Low toxicity, high efficient and overcome the resistance to chemotherapy drugs research and development has become a top priority. This research fing a compound called6-(4methyl phenyl) pyridine [2,3,6][1,4] benzene nitrogen cycloheptatriene sulphone (MPNCS) has good effect to inhibit the tumor growth also can overcome the drug resistance which mediated by the P-glycoprotein (P-gp). Base on the good effect of MPNCS anticancer effect, we though MPNCS is a potentially compound to develop for a anticancer drug.Objective1. Screening the lead compound with low toxicity and overcome drug resistance.2. In vitro studies comfirm whether the lead compound can inhibit the growth of non-small cell lung cancer cells (H460) and paclitaxol resist non-small cell lung cancer cell (H460TaxR)3. Study the Balb/c acute toxicity of the lead compound.4. Study the anti non-small cell lung cancer cells and paclitaxol resist non-small cell lung cancer cell transplant tumor model activity in the Balb/c nudemice5. Research the anti-cancer mechanism of the lead compound. Methods1. Screening the lead compound with SRB assay.2. In vitro SRB assay was used to study the anticancer actibities of MPNCS in H460and H460TaxR cell lines3. Establish the experiment of acute toxicity in Balb/c mice.4. Establish the non-small cell lung cancer transplant tumor Balb/c nudemice model, tail intravenous drug. Research the acute toxicity Balb/c and nudemice transplant tumor growth inhibition in lung cancer model.5. Preliminary research the anti-cancer mechanism: through microtube protein polymerization test, immunofluorescence test; the cell cycle and cell apoptosis detected by flow cytometry assay; through caspase-3/7, caspase-8and9test kit to assay the influence of the MPNCS to caspase kinase; testing whether MPNCS is the substrate of P-gp through the P-gp GloTM testing; the KINOMEscanTM screening the kinase targets.Results1. Screening the lead compound MPNCS with SRB assay.2. In vitro activity screening show MPNCS compound has good growth inhibition in H460and H460TaXR cells. The half growth inhibition concentration (GI50) respectively is: H460cells GI50for1.77μM, the cells to resistance for1.28μM.3. In vivo tests, acute toxicity tests proved MPNCS have lower toxicity.4. H460and H460raxR nudemice transplant tumor model proved MPNCS can inhibit the tumor growth effectively in vivo, with58.8%and55.7%tumor growth inhibition of H460and H460TaXR tumors.5. Microtubule is the target of MPNCS both in H460and H460TaxR cell and restrain the tubulin polymerize to microtubule; arrest the cell cycle at G2/M stage, followed by the cell apoptosis; kinase screening tests did not find MPNCS has kinase inhibition and MPNCS activated the caspase-8and9, which activate caspase-3the final cell apoptosis executives; MPNCS has no kinease target. ConclusionThese experiments demonstrate the compound MPNCS has effectively inhibited the growth of H460and P-gp over expression H460TaxR cells in vitro. The acute toxicity tests prove MPNCS has low toxicity in Balb/c mice and has good tumor growth inhibition to H460and H460TaxR lung cancer transplantation nudemice. From these we know MPNCS is appropriate for anticancer, also conquer the multi drug resistance. MPACS targeting the microtubule consequent on the cell cycle G2/M arrest and cell apoptosis. The way of cell apoptosis are the caspase-8and caspase-9were activated then the downstream apoptosis executive factor caspase-3was activated. Kinases screening shows there was no kinase target.