铌取代杂多钨酸盐对大鼠发育毒性的研究

发布时间：2018-04-29 12:07

本文选题：金属氧酸盐 + 致畸敏感期　；参考：《吉林大学》2013年硕士论文

【摘要】：杂多化合物，是一类前过渡金属氧阴离子簇化合物，亦是一类多核配合物。它具有高电荷、高分子量、氧化还原性强以及结构组成具有多样性等特点。杂多化合物具有广泛的用途，除了传统上用于催化、功能材料、相转移等领域，它的用途正逐渐延伸至分析化学、临床化学和药物化学等领域。在药物化学研究中，杂多化合物表现出对多种RNA病毒和部分DNA病毒具有广谱的抗病毒作用，主要体现在抗HIV、流感病毒和SARS病毒等方面，杂多化合物抗病毒研究正在引起医学界的广泛关注。本课题组在前期的研究基础上，自主设计合成了一种新型杂多化合物（代号：NCW-6），对其单晶进行了解析，明确其结构；同时研究该化合物对乙型肝炎病毒的抑制作用和一般毒性试验，结果表明，NCW-6具有良好的抗乙肝病毒的作用，作用效果优于阳性对照药物；一般毒性（急性毒性、90d喂养）结果证实，该化合物毒性较低。本研究是在前期研究工作的基础上，选用致畸敏感期试验和胎鼠中脑微团试验进一步探索NCW-6对大鼠的生殖毒性的影响，为开发具有我国自主知识产权的新型非核苷类似物药物奠定基础。本课题主要内容是通过体内生殖发育毒性试验和体外生殖毒性试验两个方面来进行评价受试药物NCW-6的生殖发育毒性。选用具有实验周期短、费用低、重现性好，便于不同实验室间的比较等特点的生殖发育毒性体外替代方法即中脑细胞微团试验进行体外毒性研究，基于大鼠受孕13d时处在神经分化前期的胚胎中脑细胞对化学毒物的作用非常敏感，细胞毒物可抑制该时期细胞分化增殖，从而使细胞集落及细胞数目减少的原理，探索外源性化学物质的致畸作用，进而从体外和体内试验综合研究受试药物的致畸作用机制。 1致畸敏感期试验应用体内生殖发育毒性试验中较重要的致畸敏感期试验来的评价受试药物NCW-6对孕鼠一般情况、胎鼠的生长发育、胚胎的形成以及对胎鼠的外观、内脏和骨骼畸形的影响等生殖发育毒性。动物在饲养两周后，随机分为5组，分别设为阴性对照组，阳性对照组，以及NCW－6高、中、低三个剂量组，剂量分别为1467.5mg/kg、366.8mg/kg、91.7mg/kg阴性对照组给予5mL/kg蒸馏水；阳性对照组给予130mg/kg的维甲酸。阴性对照组和3个给药组于各组雌鼠于受孕后第6-15天连续给药，每天固定时间灌胃一次；阳性对照组于妊娠第10天，一次灌胃给药。每3天称量体重，并根据体重变化随时调整给药量。给药期间观察动物的一般状态，，记录体重变化及死亡情况。结果显示:受试药物对孕鼠的体重和增重对照组比较差异无显著性(P0.05)；对鼠的平均黄体数、着床数、活胎数、死胎数和吸收胎数与阴性对照组比较差异不明显(P0.05)；各给药组胎鼠的平均体重、身长、尾长以及平均胎重、窝重与阴性对照组比较差异无显著性(P0.05)；各给药组胎鼠外观、内脏畸形率、骨骼畸形率与阴性对照组比较差异无显著性(P0.05)，依据新药毒理学评价方法，表明NCW-6对母体的影响、对胚胎，对胎鼠外观、内脏和骨骼畸形均无明显影响，表明在本实验条件下，NCW-6对孕鼠无母体毒性、对胎鼠无胚胎毒性及致畸作用。 2.中脑微团试验选用实验周期短、费用低、重现性好、便于不同实验室间的比较等特点的生殖发育毒性体外替代方法—中脑细胞微团试验进行体外生殖毒性研究，本实验采用13d孕鼠的胚胎中脑细胞制成密度为5×106个细胞/ml的细胞悬液分别进行细胞增殖试验和细胞分化实验，实验结果显示，受试药物IV50为1074.17μg·mL-1，ID50为394.36μg·mL-1，R值为2.72，表明该化合物生殖毒性较低。综上所述，受试药物在本研究的体外和体内生殖发育毒性试验中均表现出较低生殖发育毒性，该实验结果为NCW-6的进一步应用研究提供毒理学依据。
[Abstract]:Heteropoly compounds, a class of pre transition metal oxygen anion clusters, are also a class of polynuclear complexes. It has the characteristics of high charge, high molecular weight, strong oxidation-reduction and diversity of structure. Heteropoly compounds have a wide range of uses, except in the fields of catalysis, functional materials, phase transfer and so on. Gradually extended to analytical chemistry, clinical chemistry and pharmaceutical chemistry.
In the study of drug chemistry, heteropoly compounds show a broad spectrum of antiviral effects on a variety of RNA viruses and partial DNA viruses, mainly in the anti HIV, influenza virus and SARS virus. The study of the antivirus of heteropoly compounds is arousing widespread concern in the medical field. A new type of heteropoly compound (code name: NCW-6) was used to analyze the single crystal and to clarify its structure, and to study the inhibitory effect of the compound on hepatitis B virus and the general toxicity test. The results showed that NCW-6 had good anti HBV effect, and the effect was better than that of the positive control drug; the general toxicity (acute toxicity, 9) was better than that of the positive control drug. The results of 0d feeding confirmed that the toxicity of the compound was low. On the basis of previous research work, the effect of NCW-6 on the reproductive toxicity of rats was further explored by teratogenic sensitivity test and fetal rat midbrain micro mass test, and the foundation for the development of new non nucleoside analogues with our own intellectual property rights was established.
The main content of this topic is to evaluate the reproductive toxicity of the tested drug NCW-6 through the two aspects of the toxicity test of the body reproductive development and the test of the reproductive toxicity in vitro, and select the substitutes for reproductive development toxicity, which has the characteristics of short experimental period, low cost, good reproducibility and convenient comparison between different laboratories. The cytotoxicity study was conducted in vitro, based on the sensitivity of the embryonic mesencephalic cells in the early stage of the 13D pregnancy to the chemical toxicants. The cell toxicants could inhibit the proliferation of cells in this period, thus the principle of cell colony and cell number reduction to explore the teratogenic effect of exogenous chemicals. In vitro and in vivo experiments were conducted to study the teratogenic mechanism of the tested drugs.
1 teratogenic sensitivity test
An important teratogenic period test was used to evaluate the reproductive development toxicity of the tested drug NCW-6 on the pregnant rats, the growth and development of fetal mice, the formation of embryos, the appearance of fetal mice, the visceral and skeletal malformation of the pregnant mice. After two weeks of feeding, the animals were randomly divided into 5 groups, which were set negative. The control group, the positive control group, and the NCW 6 high, middle and low three dose groups, the dose of 1467.5mg/kg, 366.8mg/kg, 91.7mg/kg negative control group were given 5mL/kg distilled water, the positive control group was given 130mg/kg retinoic acid. The negative control group and the 3 administration group were given the female rats continuously after the pregnancy after the pregnancy. One time, the positive control group was given the medicine at tenth days of pregnancy. The weight was weighed every 3 days, and the dosage was adjusted at any time according to the weight change. The general state of the animals was observed during the period of administration and the body weight change and death were recorded. The results showed that there was no significant difference in the weight and weight gain between the pregnant rats and the control group (P0.05); The average corpus luteum number, the number of implantation, the number of live births, the number of stillbirths, the number of stillbirths, and the number of absorbed tyres were not significantly different from those of the negative control group (P0.05). The average weight, length, tail length and average fetal weight of the pregnant rats were not significant (P0.05); the appearance of the fetal rats, the rate of visceral malformation, the rate of bone malformation and the negative rate (P0.05) The sex control group had no significant difference (P0.05). According to the new drug toxicology evaluation method, it showed that the influence of NCW-6 on the mother body had no obvious influence on the embryo, the fetal mouse appearance, the visceral and bone malformation. It showed that under this experimental condition, NCW-6 had no maternal toxicity to pregnant rats, and had no embryo toxicity and teratogenic effect on fetal mice.
2. mesencephalic micro mass test
The reproductive toxicity of reproductive development toxicity in vitro, which is characterized by short experimental period, low cost, good reproducibility and convenient for comparison between different laboratories, is used to study the reproductive toxicity of the mesencephalic cell micromass test. In this experiment, the cell suspension with a density of 5 x 106 cells from the mesencephalic cells of the embryo of 13D pregnant mice was added to the cell suspension respectively. The experimental results showed that the IV50 was 1074.17 mu g / mL-1, ID50 was 394.36 mu g and mL-1, and the R value was 2.72, indicating that the reproductive toxicity of the compound was low.
In summary, the tested drugs showed low reproductive toxicity in both in vitro and in vivo reproductive toxicity tests in this study. The results provided a toxicological basis for the further application of NCW-6.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114

【参考文献】