基于数学建模和肿瘤细胞PK/PD研究中药逆转肿瘤多药耐药的体内外相关性

Reversing multidrug resistance (MDR) of tumor is an important topic at home and abroad. Traditional Chinese medicine（TCM）could be excellent MDR inhibitors as its good effect and low toxicity. But because of the complication of TCM components, the valid correlations could be hardly established between the in vitro and in vivo pharmacodynamic action(PD). Now the screening and evalution of TCM inhibitors mostly depends on pharmacodynamic trial in vivo, which is of high cost and low efficiency. More over, the usage of these TCM most rely on clinical experiences without scientific theory directing its combined use with chemotheropeutics. To solve these problems, based on the theory that the pharmacokinetics(PK) of D-luciferin potassium salt and coelenterazine in tumor cells are highly related with the "drug pump" activity of ATP-binding cassette transporters, dual-luciferase reporter assay system would be constructed combined with bioluminescence imaging technology to study PK in tumor cells of D-luciferin potassium salt and coelenterazine in vitro and in vivo. Then the PK of these two luciferases would be used as vinculum between in vitro and in vivo.The correlation of these PK parameters between in vivo and in vitro would be carried out integral analysis by canonical correlation analysis. Then nonlinear mathematical modeling method based on genetic algorithm would be selected to establishe the PK-PD model in vivo. Thus, with the correlations of PK in vitro and in vivo, the PK-PD in vivo could be predicted by PK in vitro. Using these models, systemic screening and evaluation of TCM inhibitors and optimization of its combinational usage with chemotherapeutics could be depend on in vitro trial to the maximum extent possible with high efficiency and low cost. This method would open a new mode to study TCM inhibitors reversing tumor MDR.

逆转肿瘤多药耐药（MDR）是国内外医药界的重要课题。中药逆转肿瘤MDR效果较好且毒性低。但中药成分复杂，体内外药效评价很难建立有效关联，无法通过体外预测体内，目前基本依靠体内药效试验进行逆转剂筛选与评价，效率低成本高；且临床与化疗药物联合用药方案缺乏科学的理论指导。针对以上问题，根据D-荧光素钾和腔肠素在肿瘤细胞的外排动力学（PK）与ABC转运子"药泵"活性密切相关的理论支持，通过构建双萤光素酶报告基因系统，结合活体生物发光技术，以萤光素的肿瘤细胞PK作为联系纽带，采用典型相关分析方法对体内外PK参数的相关性进行整体性分析，然后通过基于遗传算法的非线性建模法，构建具有一定信度和效度的体内PK-PD函数模型，达到从体外萤光素PK仿真预测体内PK-PD的目的，体外高效率低成本的系统筛选和评价逆转MDR的中药，优化逆转剂与化疗药物的联合用药方案。该方法开启了抗肿瘤MDR中药逆转剂的研究新模式。

逆转肿瘤多药耐药（MDR）是国内外医药界的重要课题。中药逆转肿瘤MDR效果较好且毒性低。由于中药成分复杂，体内外药效结果间很难建立有效关联，无法通过体外预测体内，目前基本依靠体内药效试验进行逆转剂筛选，效率低成本高；且临床与化疗药物联合用药方案缺乏科学的理论指导。针对以上问题，根据D-荧光素钾和腔肠素等在肿瘤细胞的外排动力学（PK）与ABC转运子“药泵”活性密切相关的理论支持，本项目以薏苡仁酯为模型中药逆转剂、以吉西他滨为化疗药物，以耐药性胰腺癌为肿瘤模型， 构建了萤火虫萤光素酶报告基因系统，结合活体生物发光技术，以萤光素的肿瘤细胞PK作为联系纽带，采用person分析方法对体内外PK参数的相关性进行分析，结果表明耐药蛋白底物D-荧光素钾在体内外肿瘤细胞的外排动力学可以作为联系体内外药效的纽带，可以以此方法进行逆转剂筛选。接下来，以榄香烯为模型中药逆转剂、以阿霉素为化疗药物，以耐药性乳腺癌为肿瘤模型，进一步验证了该方法的可靠性。将阿霉素换成分子靶向药物吉非替尼，将逆转剂榄香烯联合吉非替尼同样取得了较好的逆转效果。因此，不同逆转剂、不同肿瘤模型、不同化疗药物或分子靶向药物都证明了以肿瘤细胞的PK与ABC转运子“药泵”活性密切相关，可以作为体内外逆转肿瘤多药耐药药效相关性的联系纽带， 从而可以利用该方法， 体外高效率低成本的系统筛选逆转MDR的中药。最后，本项目选取多种中药逆转剂：葛根素、苦参碱、姜黄素、槲皮素、榄香烯、康莱特、鸦胆子油、淫羊藿苷、南蛇藤醇、大黄酸等给药后对阿霉素治疗耐药乳腺癌细胞生长抑制的影响。 最终体外选择小檗碱和槲皮素两味能逆转耐药蛋白底物外排的中药成分作为MDR逆转剂，并进一步通过体内实验进行药效实验，取得了较好的抗肿瘤效果，相比单独使用阿霉素，联合阿霉素和槲皮素、或小檗碱能在阿霉素低剂量的条件下达到和阿霉素单独使用时高剂量的相同药效，因此通过降低剂量，大大的降低了阿霉素的用药毒性并保持了良好的药效。该方法开启了抗肿瘤MDR中药逆转剂筛选的研究新模式。