基于功效成分群的中成藥生產(chǎn)全過程控制體系

肖 偉（中國(guó)藥理學(xué)會(huì)中藥與天然藥物藥理專業(yè)委員會(huì)副主任委員，江蘇康緣藥業(yè)股份有限公司研究員級(jí)高級(jí)工程師，中藥制藥過程新技術(shù)國(guó)家重點(diǎn)實(shí)驗(yàn)室，主要從事中藥新藥研究與開發(fā)。

曹 亮（世界中醫(yī)藥學(xué)會(huì)聯(lián)合會(huì)網(wǎng)絡(luò)藥理學(xué)專業(yè)委員會(huì)常務(wù)理事，江蘇康緣藥業(yè)股份有限公司高級(jí)工程師，主要從事中藥新藥研究與開發(fā)。

中藥標(biāo)準(zhǔn)化是國(guó)家重大戰(zhàn)略需求，已被納入《國(guó)家中長(zhǎng)期科學(xué)和技術(shù)發(fā)展規(guī)劃綱要》。目前，中成藥標(biāo)準(zhǔn)現(xiàn)狀與國(guó)際藥品標(biāo)準(zhǔn)水平存在較大差距，嚴(yán)重阻礙了中成藥的現(xiàn)代化、國(guó)際化進(jìn)程，突出表現(xiàn)在兩大方面，一是質(zhì)控成分相對(duì)單一，多與功效關(guān)聯(lián)性不強(qiáng)，難以保障整體功效；二是質(zhì)控成分無明確質(zhì)控范圍（有下控制限而無上控制限），生產(chǎn)過程中缺乏控制，產(chǎn)品批間波動(dòng)大，難以保證療效穩(wěn)定。導(dǎo)致上述現(xiàn)狀的重要原因有：①缺乏系統(tǒng)的功效成分發(fā)現(xiàn)模式及功效成分在中成藥系統(tǒng)下確定量效范圍的方法；②在中成藥生產(chǎn)中缺乏多成分整體控制并穩(wěn)定傳遞的技術(shù)體系。基于此，我們首先突破中藥功效成分快速確證技術(shù)，結(jié)合中成藥化學(xué)成分規(guī)?；?、快速分離制備方法，集成高通量-高內(nèi)涵活性篩選、體內(nèi)代謝和整體藥效驗(yàn)證，構(gòu)建起預(yù)測(cè)-制備-驗(yàn)證-聚焦-確證的中藥功效成分發(fā)現(xiàn)模式；其次，整合生物通路的定量計(jì)算方法和分子印跡技術(shù)，引入中藥多成分-多功效的量-效范圍研究來明確中藥功效成分的“質(zhì)量控制窗”，解決中成藥質(zhì)控范圍難以確定的問題；然后，建立基于質(zhì)量源于設(shè)計(jì)（QbD）理念的工藝辨析優(yōu)化技術(shù)和過程融合指紋圖譜技術(shù)，集成自控和在線檢測(cè)技術(shù)，形成生產(chǎn)過程中功效成分群的“各工藝點(diǎn)的點(diǎn)-點(diǎn)一致”、“各工序段的段-段一致”、“各生產(chǎn)批次的批-批一致”的質(zhì)量一致性控制體系，突破中成藥質(zhì)量均一和療效穩(wěn)定難題?！続ccording to the Outline of the National Program for Long-and Medium-Term Scientific and Technological Development,we will encourage the standardization of TCM.Moreover,the reality is that few compounds have their quality stan?dard controlled and the compounds are not attrib?uted to a certain efficacy in the most situations,and that the upper limit of the compounds is not defined,thus having negative influence on the process of TCM and posing an obstacle to global?ization of related trade and the standardization of TCM.To address these issues,some strategies have been initiated to discover active compounds and to ascertain the optimal window of the compounds at system levels.Some modern tech?nologies are employed to monitor the change of active compounds ranging from Chinese herbal medicine to preparations.Firstly,a platform is set up by integrating virtual screening,preparative chromatography,high throughputscreening,high content screening,pharamcokinetic analysis and animal models.The platform can then prepare a large number of hits and verify the effective compounds in TCM.Secondly,the reasonable range of effective compounds is defined in quality control for TCM via molecular imprinting and quantitative modeling based on pathway(QMBP).Finally,according to the concept of quality by design(QBD),a control system is built to ensure the stability from a parameter to a procedure in the production process forevery batch by combining fingerprint chromatogram,and on-line/off-line real-time quantitative monitoring.】

近年在上述研究思路的指導(dǎo)下，對(duì)源于張仲景的經(jīng)典方—桂枝茯苓方的中藥大品種桂枝茯苓膠囊進(jìn)行了基于功效成分群的全過程質(zhì)量控制體系進(jìn)行了研究，利用具有自主知識(shí)產(chǎn)權(quán)的中藥生物活性信息預(yù)測(cè)分析和制備工業(yè)色譜制備164個(gè)潛在功效成分的百克級(jí)化合物庫(kù)，通過高通量-高內(nèi)涵平臺(tái)驗(yàn)證26個(gè)功效成分，經(jīng)體內(nèi)代謝研究和整體動(dòng)物評(píng)價(jià)確證了制劑治療原發(fā)性痛經(jīng)、盆腔炎和子宮肌瘤的15個(gè)功效成分包括芍藥苷、丹皮酚、去氫茯苓酸等；借助溶膠-凝膠法制備分子印跡聚合物分別敲除制劑中相應(yīng)功效成分，整合生物通路的定量計(jì)算方法，確定各個(gè)功效成分的整體量效關(guān)系，進(jìn)而設(shè)定功效成分的“質(zhì)量控制窗”作為質(zhì)量標(biāo)準(zhǔn)制訂依據(jù)；以功效成分傳遞波動(dòng)性為指標(biāo)對(duì)工藝全程進(jìn)行質(zhì)量風(fēng)險(xiǎn)分析，確定影響質(zhì)量一致性的關(guān)鍵工序段和工藝環(huán)節(jié)，并進(jìn)一步辨識(shí)各工藝參數(shù)與質(zhì)量波動(dòng)的相關(guān)性明確法定工藝參數(shù)，之后采用近紅外在線監(jiān)測(cè)、離線快速分析，自動(dòng)化控制等技術(shù)對(duì)法定/非法定工藝參數(shù)、關(guān)鍵工序段和工藝環(huán)節(jié)進(jìn)行布局設(shè)計(jì)，達(dá)到“點(diǎn)-點(diǎn)控制”和“段-段控制”，最后建立從原料、中間體到成品的基于功效成分質(zhì)控窗的定量指紋圖譜檢測(cè)生產(chǎn)的每批制劑，累積分析質(zhì)量的均一性和療效的穩(wěn)定性，從而達(dá)到“批-批一致”。通過近3年共多批次桂枝茯苓膠囊大生產(chǎn)的驗(yàn)證，成品中各功效成分含量及指紋圖譜相似度等質(zhì)控指標(biāo)批次間RSD%由8.3%降至3.6%，明顯提高了質(zhì)量一致性，保障臨床療效穩(wěn)定，實(shí)現(xiàn)了“批-批一致”，也驗(yàn)證了該體系的科學(xué)可行?！綯ake Guizhi Fuling capsules for example.The change of the effective compounds in the process of production was explored to set up a perfect quality system.The hits,which were fished on the basis of the indication from Traditional Chinese Medicine Network Phar?macology Intelligent Information Platform,were separated up to hundreds of grams by preparative chromatography.Among the compounds,there were 26 active ingredients by high throughput screening and high content screening,15 of which(including paeoniflorin,paeonol,and dehy?dropachymic acid)were verified by identifying the components in plasma and evaluating the effect in some animal models relating to dysmenor?rhea,pelvic inflammatory disease,and endome?triosis.Then,the molecularly imprinted polymers(MIPs)of the effective compounds were synthe?tized via a sol-gel coating process.Each sample without a compound in Guizhi Fuling capsules was prepared by MIPs,and the dose-response curve was computed and tested by QMBP and in vivo for the effective compounds at the system level.The quality standard was performed in Guizhi Fuling capsules according to the optimal windows of the dose-response curve.During the produc?tion of Guizhi Fuling capsules,the key procedure and the parameters closely related to the uniform quality were defined by analyzing the fluctua?tions of the content of the effective compounds,and then were monitored by on-line/off-line nearinfrared spectroscopy and automatic control system.Finally,HPLC fingerprints were developed to quantify the effective compounds from Chinese crude drugs,intermediate products to prepara?tion for every batch,and the variation of the effective compounds was reduced from 8.3%to 3.6%.The data from three years showed that the uniformity of quality was improved,and that the products had a stable therapeutic effect in clinic.It proved that the strategy was feasible for total quality control of TCM.】