Investigation of the Mutagenic Potential of Immunomodulator Arglabin Native in Tablets in the Ames Test

The mandatory preclinical safety evaluation is an essential prerequisite to obtain the qualitative and effective medicines. Due to the fact that drugs may reveal genotoxic properties, the investigation of their mutagenic activity is an obligatory part of the preclinical drug safety program. The aim of the research is to study mutagenic properties of a new original immunomodulator Arglabin native in tablets in the induced test of gene mutations （the Ames test） on Salmonella typhimurium strains. Materials and methods： Four strains of S. typhimurium TA98, TA100, TA1535, and TA1537 were used to assess the mutagenicity in the Ames test. Results and conclusions： No statistically reliable dose-dependent increase in the number of revertant colonies of Salmonella typhimurium has been observed in the presence of the given drug within the investigated dose ranges from 5.0 to 100.0 μg/mL for strains TA100 and TA1535, and from 5.0 to 250 μg/mL for strains TA98 and TA1537 against the baseline of spantaneous mutations. Arglabin native in tablets does not reveal a mutagenic activity within the studied dose ranges on Salmonella typhimurium strains TA98, TA100, TA1535, TA1537.

HPLC法测定二甲氨基阿格拉宾盐酸盐的含量

目的:建立二甲氨基阿格拉宾盐酸盐的含量测定方法。方法:色谱柱为氨基柱(250mm×4.6mm,5μm),以乙腈-0.01mol·L^(-1)磷酸铵(用磷酸调节pH至6.5)(60:40)为流动相,流速为1.0ml·min^(-1),检测波长为210nm,柱温30℃,进样量20μl。结果:二甲氨基阿格拉宾盐酸盐与阿格拉宾及其他杂质之间能够达到很好分离;二甲氨基阿格拉宾盐酸盐在1.0~6.0mg·ml^(-1)浓度范围內线性关系良好(r=0.9999),平均加样回收率为99.07%,RSD为0.76%(n=9)。结论:该法测定二甲氨基阿格拉宾盐酸盐含量,简便、准确、重复性好。

阿格拉宾对小鼠肺缺血再灌注损伤的保护作用及机制

目的观察阿格拉宾在小鼠肺缺血再灌注损伤中的作用并探讨可能机制。方法根据随机数表法将40只8~10周龄雄性C57/B6小鼠随机分为4组,即对照组、阿格拉宾组、缺血再灌注组和治疗组。采用左肺门夹闭1h,再灌注2h的方式构建小鼠肺缺血再灌注损伤模型,治疗组小鼠在肺门夹闭前1h给予阿格拉宾(5μg/kg)腹腔注射。再灌注结束后,留取左肺组织行HE染色评估肺损伤状况,同时Western blot法检测炎性因子[白细胞介素-1β(interleukin-1β,IL-1β),单核细胞趋化因子-1(monocyte chemoattractant protein-1,MCP-1),肿瘤坏死因子(tumor necrosis factor,TNF)-α]、焦亡标志物[NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)和凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a caspase-recruitment domain,ASC)]及自噬相关蛋白(Beclin1和ATG7)的表达。结果缺血再灌注组较对照组小鼠肺组织病理损伤明显加重,肺组织炎性因子IL-1β、MCP-1和TNF-α相对表达明显升高(P均<0.05),焦亡相关标志蛋白NLPR3和ASC的表达水平明显升高(P均<0.05),同时自噬相关蛋白Beclin1和ATG7被明显抑制(P均<0.05);予以阿格拉宾预处理后,肺缺血再灌注小鼠肺病理损伤明显减轻、肺组织炎性因子IL-1β、MCP-1和TNF-α相对表达明显降低(P均<0.05);焦亡相关标志蛋白NLPR3和ASC的表达水平明显降低(P均<0.05),自噬相关蛋白Beclin-1和ATG7被明显上调(P均<0.05)。结论阿格拉宾可能通过抑制焦亡并激活自噬改善小鼠肺缺血再灌注损伤。

HPLC法测定多花蒿药材中阿格拉宾含量

目的:建立用高效液相色谱法测定多花蒿药材中阿格拉宾含量。方法:采用回流提取法,以甲醇为提取溶剂。色谱柱为Agilent NH_2(250 mm×4.6 mm,5μm),以乙腈-水(60:40)为流动相,流速为1.0 ml·min^(-1),检测波长为210 nm,柱温30℃。结果:阿格拉宾与其它杂质能够达到有效分离,在0.5～499.0μg·ml^(-1)浓度范围内线性关系良好(r=0.999 9);平均加样回收率为99.5%,RSD为0.6%(n=9)。结论:该检测方法准确可靠,重复性好,可用于批量多花蒿药材中阿格拉宾含量的测定。

高速逆流色谱法从多花蒿中分离制备阿格拉宾

目的采用高速逆流色谱法分离制备多花蒿中阿格拉宾。方法多花蒿乙醇提取物经硅胶柱色谱分离,所得组份Fr.1用高速逆流色谱进一步分离,采用正己烷-醋酸乙酯-甲醇-水(1.2:0.8:1.5:0.5)为两相溶剂系统,上相为固定相,下相为流动相,主机转速850r/min,体积流量2.0mL/min,检测波长为254nm。采用波谱法对所得化合物进行结构鉴定,HPLC法测定产品的纯度。结果从500mg组分Fr.1中得到260mg质量分数为99.5%的阿格拉宾。结论该方法操作简单,可用于阿格拉宾化学对照品的分离制备。