Preparation Of Combination Drug HAMPT Towards Cancer Metastasis Prevention And Its Related Study In Pharmacodynamics And Toxicology

Purpose:The emergence of metastases remains the primary cause of mortality in cancer patients and accounts for nearly 90%of cancer patient deaths.Current chemotherapeutics that are originally designed to kill high proliferating carcinoma cells are ineffective in targeting low proliferating CTCs in the blood of cancer survivors.Combined with our newly-established "kinetics of cancer metastasis",we designed,accordingly,a quadruple combination drug HAMPT(standing for highly active metastasis prevention therapy),which is consisted of mifepristone(RU486),aspirin,lysine and doxycycline.The aim of the article is to study the pharmacodynamics and toxicology of HAMPT and clarify the mechanisms of prevention of cancer metastasis.Methods:The HPLC and UPLC-MS/MS methods were established to determine if there are chemical drug-drug interactions among HAMPT active ingredients.The MTT and adhesion assays were conducted to evaluate cytotoxicity and anti-adhesion of HAMPT.Flow cytometry was used to characterize effects of HAMPT on expression of cell adhesion molecules,cell cycle,and platelet-induced cell aggregation.The cell scratch test was conducted to test effect of HAMPT on cell migration.The in vivo chemopreventive effect of HAMPT was observed in mouse metastasis model.Fourteen-day of acute toxicity assay and fifty-day sub-acute toxicity assay were established to detect toxicity of HAMPT to show its difference from traditional cancer drug and can be taken long by patients.Results:The quantitative analysis revealed no chemical drug-drug interactions among HAMPT active ingredients.HAMPT had marginal cytostatic effect on cells at concentrations up to 100 ?g/mL.Whereas,HAMPT caused a significant dose-dependent inhibition(1-50 ?g/mL)on adhesion and implantation of caner cells to HUVECs and Matrigel.Flow cytometry analysis revealed that HAMPT inhibited IL-1 ? induced expression of ICAM-1 and E-selectin on HUVECs,?4 integrin on B16-F10 as well as Sialy-Lewis X on HT-29 cells.The in vivo experiment showed that HAMPT suppressed B16-F10 and CT-26 metastasis to mouse lungs in a dose-dependent manner,resulting in reduced number of lung nodules and weight in the metastatic model.As a result,lung weight of the treated mice kept within the normal range.The combination also showed better effect comapred with other combinations among four drugs.Fifty-day rat toxicity study with HAMPT at doses(670-1340 mg/kg)20-fold higher than its therapeutic dose produced no significant toxicity.Interestingly,the acute toxic death could not be reached at the maximum administrable dose(5 g/kg).Conclusion:HAMPT could safely prevent cancer metastasis in mechanisms by which it improves the microenvironment to inhibit CTC circulating and causes significant inhibitory effects on adhesion of cancer cells to vascular endothelium of distant tissue.In addition to its comprehensive inhibition on platelet aggregation,the present study provides a stronger scientific basis for proposing HAMPT for global cancer metastatic chemoprevention.