【摘要】：目的:通過(guò)微量富集和模糊衍生方法初步建立抗結(jié)核新型藥物篩選平臺(tái),并從已知抗結(jié)核藥物中篩選新的活性成分。方法:將異煙肼(INH)、利福平(RFP)、乙胺丁醇(EMB)、鏈霉素(SM)、卡那霉素(KM)、阿米卡星(AMK)、氧氟沙星(Ofx)、左氧氟沙星(Lfx)、加替沙星(Gfx)、莫西沙星(Mfx)、對(duì)氨基水楊酸鈉(PAS)和丙硫異煙胺(Pto)共12種抗結(jié)核藥物進(jìn)行模糊衍生,并對(duì)衍生后產(chǎn)生的新成分進(jìn)行微量富集篩選抗結(jié)核新成分。將上述藥品分別配制成0.5%的甲醇溶液,每種藥品溶液以線條狀上樣在一張薄層板上,并在薄層板上進(jìn)行模糊衍生。將衍生的混合物進(jìn)行二維薄層色譜(2D-TLC)展開(kāi),在第一個(gè)維度將藥品衍生物中的成分根據(jù)不同極性進(jìn)行分離,形成條狀譜帶,然后將薄層板平行旋轉(zhuǎn)90°進(jìn)行二維展開(kāi),把條狀譜帶推至薄層板邊緣并聚集為點(diǎn)狀譜帶,達(dá)到微量成分富集的作用。將點(diǎn)狀譜帶逐一分割,分別置于鋪滿結(jié)核分枝桿菌(MTB)的培養(yǎng)基上,37°C恒溫培養(yǎng)箱培養(yǎng)4-8周,觀察各譜帶的抑菌作用。結(jié)果:INH和EMB兩組中存在相應(yīng)譜帶區(qū)域有抑制MTB生長(zhǎng)的作用,其中INH組的抑菌實(shí)驗(yàn)的靈敏度和特異度較高,進(jìn)一步分割I(lǐng)NH(INH對(duì)照組)和INH I(INH衍生物組)的抑菌區(qū)域逐一譜帶進(jìn)行培養(yǎng),從而得出INH I(4號(hào)譜帶)具有顯著抑制敏感結(jié)核分枝桿菌和耐藥結(jié)核分枝桿菌生長(zhǎng)的作用,INH I組中可能產(chǎn)生了新成分。結(jié)論:初步建立了微量富集方法和模糊衍生技術(shù)篩選抗結(jié)核新藥的平臺(tái),使數(shù)個(gè)實(shí)驗(yàn)室工作在同一張薄層板上進(jìn)行,從而具有高效、低成本和省時(shí)省力等優(yōu)點(diǎn)。本技術(shù)可把已知抗結(jié)核藥物中的微量成分分離出來(lái),通過(guò)體外抑菌實(shí)驗(yàn),從已知的藥物中發(fā)現(xiàn)抗結(jié)核新成分。在此平臺(tái)基礎(chǔ)上,初步從INH的衍生物中篩選出抗結(jié)核新成分,為藥物的分離、篩選及新藥的研發(fā)奠定了基礎(chǔ)。
[Abstract]:Objective: to establish a new anti-tuberculosis drug screening platform by microenrichment and fuzzy derivation, and to screen new active components from known anti-tuberculosis drugs. Methods: twelve species of isoniazid (INH), rifampicin (RFP), (RFP), rifampicin (RFP), (EMB), streptomycin (SM), (KM), (AMK), ofloxacin (Ofx), levofloxacin (Lfx), gatifloxacin (Gfx), moxifloxacin (PAS) and (Pto) were prepared. Antituberculotic drugs are derived by fuzzy methods, New anti-tuberculosis components were obtained by microenrichment and screening. The above drugs were prepared into 0.5% methanol solution, and each drug solution was taken as a linear strip on a thin plate, and fuzzy derivation was carried out on the thin layer plate. The derivative mixture was expanded by two-dimensional thin-layer chromatography (2D-TLC), the components of the derivative were separated according to different polarities in the first dimension to form a strip, and then the thin plate was rotated 90 擄parallel for two-dimensional expansion. The strip band was pushed to the edge of the thin plate and aggregated into a dot band, which resulted in the enrichment of trace components. The dot bands were divided one by one and cultured in a 37 擄C constant temperature incubator for 4-8 weeks on a culture medium covered with Mycobacterium tuberculosis (MTB). The bacteriostatic effects of each band were observed. Results in the two groups, the corresponding band region could inhibit the growth of MTB. The sensitivity and specificity of the bacteriostatic test in the INH group were higher than those in the control group. Further partitioning the INH (INH control group and the INH I (INH derivative group), the inhibitory regions were cultured one by one, It is concluded that INH I (4) has a significant inhibitory effect on the growth of sensitive Mycobacterium tuberculosis and drug-resistant Mycobacterium tuberculosis. Conclusion: a platform for screening new anti-tuberculosis drugs by microenrichment method and fuzzy derivation technique has been established preliminarily, which makes several laboratories work on the same thin-layer plate, which has the advantages of high efficiency, low cost and time saving and labor saving. This technique can isolate the trace components from known antituberculous drugs and find new antituberculotic components from known antituberculous drugs by in vitro bacteriostasis test. On the basis of this platform, a new anti-tuberculosis component was screened from the derivatives of INH, which laid a foundation for the separation and screening of drugs and the research and development of new drugs.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R52

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉焱,趙詠,楊輝;薄層色譜法鑒別去痛片四種主要成分[J];錦州醫(yī)學(xué)院學(xué)報(bào);2003年04期

2 解翠珠;;薄層色譜法鑒別硫酸慶大霉素注射液的改進(jìn)[J];華西藥學(xué)雜志;2006年04期

3 單紅艷;;薄層色譜法檢驗(yàn)中藥應(yīng)注意的問(wèn)題[J];中國(guó)中醫(yī)藥現(xiàn)代遠(yuǎn)程教育;2011年17期

4 T.A.Vinson;何萍雯;;篩選尿中巴比妥鹽的靈敏薄層色譜法(摘譯)[J];刑事技術(shù);1982年03期

5 敖利;薄層色譜法定性鑒別參麥凍干粉針中糖的組成[J];中國(guó)藥業(yè);2004年08期

6 戴衛(wèi)紅;李云霞;張素芬;;軟肝升白顆粒的薄層色譜法鑒別[J];中國(guó)藥師;2009年03期

7 郝艷玲;;痤瘡顆粒的薄層色譜法定性研究[J];中國(guó)民族民間醫(yī)藥;2009年08期

8 具善姬;張明子;;薄層色譜法鑒別胃康樂(lè)顆粒[J];中國(guó)藥業(yè);2012年14期

9 顏曉航;;薄層色譜法操作技術(shù)控制要點(diǎn)分析[J];安徽醫(yī)藥;2012年09期

10 姚桂根,郭濟(jì)賢;棒狀薄層色譜法及其在天然藥物分析中的應(yīng)用[J];中成藥;1988年08期

相關(guān)會(huì)議論文 前10條

1 唐洪梅;閆雪;李得堂;;微乳薄層色譜法分離鑒定腸激安方中多種有效成分[A];2010年廣東省藥師周大會(huì)論文集[C];2011年

2 宋家玉;周景洋;豐碩;陳金東;邢金川;于水永;;薄層色譜法檢測(cè)降糖中藥制品中的格列苯脲[A];山東預(yù)防醫(yī)學(xué)會(huì)第二屆學(xué)術(shù)年會(huì)論文匯編[C];2006年

3 王淑娟;劉春峰;孫偉;;薄層色譜法鑒別通脈顆粒[A];2010年度全國(guó)醫(yī)藥學(xué)術(shù)論文交流會(huì)暨臨床藥學(xué)與藥學(xué)服務(wù)研究進(jìn)展培訓(xùn)班論文集[C];2010年

4 錢麗華;申蘭慧;許園園;秦臻臻;;薄層色譜法檢測(cè)安眠類中藥中非法添加的地西泮[A];2006年全國(guó)藥物質(zhì)量分析學(xué)術(shù)研討會(huì)——《藥物分析雜志》第二屆普析通用杯優(yōu)秀論文評(píng)選頒獎(jiǎng)會(huì)論文集[C];2006年

5 李繼民;王彥吉;鄒寧;陳文勝;;薄層色譜法測(cè)定紙張水解液中糖的初步研究[A];中國(guó)化學(xué)會(huì)第二十五屆學(xué)術(shù)年會(huì)論文摘要集（下冊(cè)）[C];2006年

6 劉曉涵;陳永剛;林勵(lì);肖鳳霞;趙紅英;林芳花;;薄層色譜法分析DFMA[A];全國(guó)第8屆天然藥物資源學(xué)術(shù)研討會(huì)論文集[C];2008年

7 喬化民;喬艷;蔡林;吳雪;楊素葉;;薄層色譜法檢測(cè)排石沖劑的質(zhì)量[A];中華中醫(yī)藥學(xué)會(huì)第九屆中藥鑒定學(xué)術(shù)會(huì)議論文集——祝賀中華中醫(yī)藥學(xué)會(huì)中藥鑒定分會(huì)成立二十周年[C];2008年

8 喬化民;喬艷;蔡林;吳雪;楊素葉;;薄層色譜法檢測(cè)排石沖劑的質(zhì)量[A];《醫(yī)藥導(dǎo)報(bào)》第八屆編委會(huì)成立大會(huì)暨2009年度全國(guó)醫(yī)藥學(xué)術(shù)交流會(huì)和臨床藥學(xué)與藥學(xué)服務(wù)研究進(jìn)展培訓(xùn)班論文集[C];2009年

9 章江生;管佳;李紹平;;4種姜黃屬藥材的二次展開(kāi)薄層色譜法定性定量分析[A];2008年《藥物分析雜志》第三屆普析通用杯論文集[C];2008年

10 張華蕓;孫海燕;郭霞;;康鼻膠囊的研制與質(zhì)量控制[A];2006年山東省醫(yī)院藥學(xué)學(xué)術(shù)研討會(huì)論文匯編[C];2006年

相關(guān)重要報(bào)紙文章 前1條

1 江蘇連云港市東海藥監(jiān)局 莊友田;薄層色譜法的應(yīng)用與操作[N];中國(guó)醫(yī)藥報(bào);2003年

相關(guān)碩士學(xué)位論文 前5條

1 鄭超;芪芍活血顆粒的質(zhì)量標(biāo)準(zhǔn)研究[D];河南中醫(yī)藥大學(xué);2016年

2 岳健博;微量富集及模糊衍生技術(shù)篩選抗結(jié)核新活性成分[D];遵義醫(yī)學(xué)院;2017年

3 張婧;薄層色譜法和高效液相色譜法綜合考察簽字筆油墨[D];首都師范大學(xué);2007年

4 何曉晗;薄層色譜法檢測(cè)果蔬中仲丁胺殘留量技術(shù)研究[D];沈陽(yáng)農(nóng)業(yè)大學(xué);2004年

5 陳冬雪;羚竺散的質(zhì)量標(biāo)準(zhǔn)研究[D];吉林大學(xué);2007年

，

本文編號(hào)：2248858