The Distribution In Kunming Rats And The Permeability On Caco-2Cell Model Of MINS, An Antitumor Compound

Polyamines (putrescine, spermidine, spermine) exert an important role in cell proliferation,differentiation, ulcer healing, etc. Previous studies demonstrated that tumor cells demand much morepolyamines than normal cells for growth. Meanwhile, polyamines transport system of tumor cellmembrance further enhanced than normal cells for uptake the exogenous polyamine. Natural polyamines orsynthetic polyamines graft with cytotoxic drugs, the conjugates could be recognized by polyaminestransport system and also uptaked into the tumor cells.MINS, a novel naphthalimide-polyamine conjugate, exerts potent antitumor activity both in vitroand in vivo. In this article, high-performance liquid chromatographic (HPLC) method is used to detect thetissue distribution and serum concentration of MINS in Kunming mice. The detection method of polyamine(putrescine, spermidine, spermine) level in mice’s tissue and serum is same as above.The extractingsolution of MINS was monitored by fluorescence detection(excitation:225nm; emission:745nm)．MINSwas separated on BDS-C18column (250×4.6mm) and eluted with methanol and distilled water (methanol:distilled water=58:42). The flow rate was1mL/min and the temperature was20℃. The retention time ofMINS is about7min and the low of detection is0.5ng/mL. While the samples were eluted for polyamineseparation with a linear gradient starting from65%methanol to100%and water starting from35%to0%within25min on XDB-C18column (250×4.6mm) by fluorescence detector achieving excitation at340nmand measuring its emission at515nm. In this method, hexamethylenediamine is used as internalstandard,and the detection limit were0.19pmol,0.20pmol,0.23pmol respectively. Results show thatMINS impacts on organisations polyamine content in mice. According to the results, the main metabolismand excertion organs of MINS in mice is liver and kidney.Experimental animals: Kunming mice were purchased from the experimental animal center ofHenan. Mice were divided into two group: normal group and liver damage group. The mode of mice liverdamage modulus method: use0.6%carbon tetrachloride by intraperitoneal injection of peanut oil solutionto0.1mL/10g. The distribution of MINS in tissues and organs was detected by HPLC through tail veininjection at the dose of2mg/kg. Mice were then killed by cervical dislocation after taking blood sample, whereafter heart, liver, spleen, lung, kidney and brain were dissection and washed with saline, then storedat80°C until processed for HPLC use. The blood sample was centrifuged (3000r/min) after standing2hat4℃, then taken supernatant and stored at80℃until use. The concentration of MINS in serum andtissues of normal mice at the point of1h is0.7677,0.7830,0.6980,3.4915,2.1657,0g/g, and0.2201,0,0.4445,6.0735,3.5431,2.3639,0.4814g/g of liver damage mice, respectively.The absorption of MINS were evaluated in Caco-2monolayer cell model which forms confluentmonolayers and differentiates to cells with an enterocyte-like morphology using standard cell cultureconditions. Moreover, it represents a valuable model system for the investigation of drug transport acrossthe small intestinal epithelium. MINS (40μM,50μM,60μM) was dispersed in apical transport medium,then took sample after30min,1h,1.5h,2h,3h,4h. MINS and polyamine in samples were detected byHPLC. The results show that the quantity of MINS and polyamine is diversity at different point.The resultsshow that the MINS concentration of50μM is the best absorption concentration.