酚噻嗪类光敏剂YWW007用于病原体灭活的毒性评价研究

The genotoxicity study of YWW007,a new photosensitizer derived from phenothiazine

目的探讨酚噻嗪类光敏剂YWW007应用于血液病原体灭活的安全使用剂量。方法细胞毒性试验:使用YWW007终浓度为1、4、12、20、40μmol/L培养基培养小鼠成纤维细胞,用四甲基偶氮唑盐比色(MTT)方法检测细胞存活率;遗传毒性评价:采用YWW007终浓度为1、2、4、6、8、12μmol/L的培养基培养小鼠淋巴瘤细胞,用微孔板法做tk基因突变试验,计算平板效率、相对存活率以及TFT抗性突变频率等指标。结果当YWW007浓度达到40μmol/L时,具有明显的细胞毒性;而YWW007浓度为12、4、2和1μmol/L时,细胞存活率>70%,提示细胞毒性轻微;YWW007浓度≤4μmol/L时,在含有和不含有代谢活化系统(S9)条件下,其突变频率(MF)值均未达到阴性对照组MF值的2倍,未见剂量-效应关系。结论 YWW007浓度≤4μmol/L为用于病原体灭活的安全剂量。

Objective To evaluate the genotoxieity of YWW007, a phenothiazine derivative, and provide safety margin for application of photosensitizable YWW007. Methods YWW007 was purified before tests. The evaluation of in vitro eytotoxieity was based on the viability of mouse fibroblast cells incubated with 1,2, 4, 20 and 40μmol/L of YWW007. The survival rate of cells was measured by thiazolyl blue tetrazolium bromide method. For genotoxicity evaluation, L5178 TK^＋/- mouse lymphoma cells were incubated with 1,2, 4, 6, 8μmol/ L of YWW007 with or without the presence of metabolic system （S9）. The mutation frequency （MF） was determined by measuring the mutation rate of thymidine kinase gene （TK）. Results At the dose of 40uM, cytotoxicity of YWW007 was significant while at the dose of 12μmol/L, 4μmol/L, 2μmol/L and 1μmol/L, cytotoxicity was slight. The negative gene mutation results of YWW007 were seen at the dose of 4μmol/L, 2μmol/L and 1μmol/L, as in the presence or absence of S9, MF did not go beyond twice of the MF value of negative control, and no dose-effect relationship was observed. Conclusion The safe applicable concentration of YWW007 for inactivation is ≤4μmol/L.