在國際無菌制藥行業(yè)，有2部關鍵法規(guī)和指南被尊為圣經(jīng)，其中一部是FDA發(fā)布的無菌工藝指南，另外一部就是EU GMP附錄1。經(jīng)過漫長的修訂期，在2022年8月，EU GMP附錄1修改工作終于結(jié)束，定稿版本發(fā)布。這份新的GMP指南影響很大，因此為了讓企業(yè)有適應期，設定生效日期是2023年8月。

       在近期，PIC/S也根據(jù)已經(jīng)發(fā)布的EU GMP附錄1發(fā)布了自己的GMP指南的附錄1。而中國已經(jīng)在2021年提出加入PIC/S的申請，可以預見的是，EU GMP附錄1對中國制藥行業(yè)的影響也會不斷加強。

       為了幫助國內(nèi)無菌企業(yè)更快的熟悉EU GMP附錄1的具體要求，并加強擬進入國際市場的無菌制藥企業(yè)的質(zhì)量提升工作，特梳理和分析EU GMP附錄1中的強制性條款，幫助企業(yè)技術(shù)人員更快理解和熟悉這份關鍵法規(guī)的要求。

       說明：

       1- 下面為了行文流暢，從EU GMP附錄1中摘引的原文，只是引用強制性和要求非常具體的段落；無關內(nèi)容沒有被引用。

       2- 下面內(nèi)容只是引用了部分EU GMP附錄1的內(nèi)容，不是全部內(nèi)容。

       第一部分：廠房設施部分的強制性條款

       4.4-The maintenance of unidirectional airflow should be demonstrated and qualified across the whole of the grade A area.

       解析：應該在整個A級區(qū)域內(nèi)部證明和確認對單向流的維護。通過這句話可以看出，A級區(qū)域的內(nèi)部驗證和測試不能只測試某個位置，應該基于A級區(qū)域面積和A級區(qū)域內(nèi)部的關鍵工位的評估，來選擇足夠測試點，來證明A級區(qū)域內(nèi)部的單向流。

       4.4-Air pressure differences should be continuously monitored.

       解析：應該對壓差進行持續(xù)監(jiān)控。從這句可以看出，應該對關鍵位置壓差，進行持續(xù)監(jiān)控。這些關鍵位置的壓差指的是A級-B級區(qū)域之間的壓差，以及B級-C級區(qū)域之前的壓差。

       4.6-Sliding doors may be undesirable for this reason.

       解析：在無菌車間不能使用推拉門，也不能使用懸吊門。因為這樣類型的門，存在更多不易清潔的死角。

       4.9-Sinks and drains should be prohibited in the grade A and grade B areas.

       解析：在A級和B級區(qū)域不能出現(xiàn)水池和地漏。這個問題，不需要解釋，這是GMP基本原則和無菌制藥特征決定的。

       4.10-The transfer of materials, equipment, and components into the grade A or B areas should be carried out via a unidirectional process. The removal of items from the grade A and B areas (e.g. materials, waste, environmental samples) should be carried out via a separate unidirectional process.

       解析：將物料、設備和部件傳遞進入A級區(qū)域或者B級區(qū)域，應該采用單向流程。將物料、廢棄物、環(huán)境監(jiān)控樣品從A級區(qū)域和B級區(qū)域轉(zhuǎn)移出來時，應該從一個單獨的單向流通道轉(zhuǎn)移出來。從這句要求可以看出，進入A/B級區(qū)域的通道應該單獨設置，進入通道和生產(chǎn)廢棄物退出通道，應該分開。

       4.11-The final stage of the airlock should, in the "at rest" state, be of the same cleanliness grade (viable and total particle) as the cleanroom into which it leads.

       解析：人流物流通道最后一個階段的房間，應該和擬進入?yún)^(qū)域在靜態(tài)下保持相同的標準（微生物項目和懸浮粒子項目）。

       4.12-Personnel airlocks: Areas of increasing cleanliness used for entry of personnel (e.g. from the grade D area to the grade C area to the grade B area). In general hand washing facilities should be provided only in the first stage of the changing room and not be present in changing rooms directly accessing the grade B area.

       解析：人流通道氣鎖房間：這些氣鎖房間是用于人員進入的房間，采用逐級提升潔凈度的方式來實現(xiàn)人員進入（例如從D級進入C級，然后再進入B級）。通常，洗手設施應該僅僅在更衣室通道的第一個房間設置，而不應該出現(xiàn)在緊靠B級區(qū)域的房間設置洗手裝置。

       從上面這段要求，相信制藥企業(yè)明白無菌車間更衣通道設置的一般性要求。雖然措辭沒有使用必須，但是歐盟GMP附錄1采用舉例的方式告訴制藥行業(yè)，這是最常見和最推薦的方式之一。

       4.12-Only materials and equipment that have been included on an approved list and assessed during validation of the transfer process should be transferred into the grade A or grade B areas via an airlock or pass-through hatches.

       解析：只有納入批準清單的物料和設備才可以通過傳遞窗或者緩沖間傳遞進入A級或者B級區(qū)域。從這句可以看出，凡是進入A級或者B級區(qū)域的物料和設備，必須納入一個特定的批準目錄中。

       4.13-For pass-through hatches and airlocks (for material and personnel), the entry and exit doors should not be opened simultaneously.

       解析：對于人員和物料進出的傳遞窗和緩沖間，進出的門不能同時打開。這是為了控制污染提出的基本要求，不需要過多解釋。

       4.14-Adjacent rooms of different grades should have an air pressure difference of a minimum of 10 Pascals (guidance value).

       解析：潔凈區(qū)內(nèi)部不同級別相鄰房間之間的壓差應該保持最低10Pa（指導值）。從這條可以看出，具體保持多大的壓差數(shù)據(jù)，需要結(jié)合更具體的QRM來判斷，而不是一律保持10Pa就算可以。

       4.15 Airflow patterns within cleanrooms and zones should be visualised to demonstrate that there is no ingress from lower grade to higher grade areas and that air does not travel from less clean areas (such as the floor) or over operators or equipment that may transfer contamination to the higher grade areas.

       解析：應該對潔凈區(qū)和潔凈室的氣流模式進行可視化研究，以證明沒有空氣從低級別區(qū)域向高級別區(qū)域侵入，以及空氣不會從較低潔凈區(qū)域（例如地面）攜帶污染物越過人員或者設備向較高等級區(qū)域傳遞。

       從這句要求看，潔凈區(qū)內(nèi)部送風、回風、排風設置要非常精細的設計才可以實現(xiàn)這樣的工藝目的。而且驗證要求也提到很高程度，需要證明這些風險是不存在的。

       4.19-Negative pressure isolators should only be used when containment of the product is considered essential (e.g. radiopharmaceutical products) and specialized risk control measures should be applied to ensure the critical zone is not compromised.

       解析：當產(chǎn)品的污染風險（例如放 射 性藥品）被認為是基本問題時，才允許使用負壓隔離器，采用采用特定的風險控制措施，以確保關鍵區(qū)域不被污染。這句話描述了負壓隔離器使用的特定情況。

       4.20-The background environment for open isolators should generally correspond to a minimum of grade C. The background for closed isolators should correspond to a minimum of grade D.

       解析：O-隔離器的背景環(huán)境通常最低是C級。而C-隔離器的背景環(huán)境通常最低是D級。這句描述了在為隔離器設置背景環(huán)境是的特定要求。

       4.21-Isolators-Generally glove integrity testing should be performed at a minimum frequency of the beginning and end of each batch or campaign.

       解析：對于隔離器上面的手套完整性測試最低頻率是每個批次或者每個生產(chǎn)周期的開始和結(jié)束，各測試一次。

       RABS-Gloves should be visually examined with each use, and integrity testing should be performed at periodic intervals.

       解析：對于RABS，應該每次使用是目視檢查手套的完整性，并且應該在周期性間隔測試手套完整性。

       4.30-Unidirectional airflow systems should provide a homogeneous air speed in a range of 0.36 - 0.54 m/s (guidance value) at the working position, unless otherwise scientifically justified in the CCS. Airflow visualization studies should correlate with the air speed measurement.

       解析：在工作位置，單向流系統(tǒng)應該提供均一的空氣風速，范圍是0.36 - 0.54 m/s，除非在CCS中對于風速另有論證。氣流可視化研究應該和風速測量相互關聯(lián)。

       通過上面這段描述，我們知道，具體單向流風速要求，應該和工藝和具體位置相關，而不是必須采用0.36 - 0.54 m/s。

       4.32-The maximum time interval for requalification of grade A & B areas, is 6 months. The maximum time interval for requalification of grade C & D areas, is 12 months.

       解析：A/B級區(qū)域的再確認周期最短是6個月。C/D級區(qū)域再確認周期最短是12個月。上面描述無菌制藥企業(yè)對于潔凈區(qū)再確認的最低周期要求。

       4.35 Disinfectants and detergents used in grade A and grade B areas should be sterile prior to use.

       解析：用于A級和B級區(qū)域的消毒劑和清潔劑應該在使用前滅菌或者除菌。從這條可以看出，A/B級是無菌區(qū)域，必須嚴格控制微生物污染。

       第二部分：設備部分的強制性條款

       5.2-The frequency at which alarms are assessed should be based on their criticality (with critical alarms reviewed immediately).

       解析：對于報警信息的審核應該基于它們的關鍵程度（關鍵報警信息必須立即審核）。從這段可以看出，如果制藥公司對于某些報警信息列為關鍵信息，應該安排人員24小時值班來處理。

       5.8-A conveyor belt should not pass through a partition between a grade A or B area and a processing area of lower air cleanliness, unless the belt itself is continually sterilised (e.g. in a sterilising tunnel).

       解析：傳送帶不能在A級區(qū)域之外運行，或者不能在B級和較低級別區(qū)域之間運行，除非傳送帶自身可以持續(xù)滅菌，例如隧道烘箱。

       這句告訴我們，在設置傳送帶的時候，必須遵守這些基本原則。

       5.9-Tube length should typically be no longer than 1m unless justified and the number of bends should be minimized.

       解析：懸浮粒子計數(shù)器取樣管長度通常不能超過1米，除非另有說明。而且，取樣管上面的彎曲數(shù)量應該最小化。這個要求應該是行業(yè)共識了，請牢記山東省某企業(yè)被歐盟警告的案例。

       第三部分：公用系統(tǒng)部分的強制性條款

       6.10 Water for injections (WFI) should be produced from water meeting specifications that have been defined during the qualification process, stored and distributed in a manner which minimizes the risk of microbial growth (e.g. by constant circulation at a temperature above 70°C).

       解析：注射用水 (WFI) 應使用符合在確認過程中定義的標準的水生產(chǎn)，以將微生物生長風險降至最低的方式儲存和分配（例如在高于 70°C 的溫度下持續(xù)循環(huán)）。從這條可以看出，WFI循環(huán)溫度是硬性要求。

       6.12-Chemical testing results should be approved before the water system is returned to use and microbiological/endotoxin results verified to be within specification and approved before batches manufactured using water from the system are considered for certification/release.

       解析：在水系統(tǒng)恢復使用之前，應該對于化學測試結(jié)果進行審核批準；使用相關工藝用水生產(chǎn)的藥品在放行之前，微生物項目和內(nèi)毒素項目應該被確認符合標準并得到批準。從這條要求看，應該對于WFI進行批次劃分，在生產(chǎn)記錄上面記錄生產(chǎn)使用的WFI批號，并按照規(guī)定完成放行。

       6.13-Sample plans should be based on the qualification data, should consider the potential worst case sampling locations and should ensure that at least one representative sample is included every day of the water that is used for manufacturing processes.

       解析：對于工藝用水的取樣計劃應該基于確認數(shù)據(jù)，應該考慮潛在的最差取樣位置，應該確保每天至少取樣一個代表性的取樣點，來評估用于生產(chǎn)的工藝用水的質(zhì)量。根據(jù)上面這段要求，對于水系統(tǒng)取樣計劃，應該采用常見的3+N或者4+N方法。

       6.15 WFI systems should include continuous monitoring systems such as Total Organic Carbon (TOC) and conductivity, as these may give a better indication of overall system performance than discrete sampling.

       解析：應該對于WFI系統(tǒng)采用持續(xù)監(jiān)控，測試TOC和電導率項目，因為這樣可以及時反饋整個系統(tǒng)的性能，比離線測試要更好。從這句看，對于WFI系統(tǒng)，在線監(jiān)控TOC和電導率是必須項目。

       第四部分：人員管理部分的強制性條款

       7.5 Unqualified personnel should not enter grade B cleanrooms or grade A in operation. If needed in exceptional cases, manufacturers should establish written procedures outlining the process by which unqualified personnel are brought into the grade B and A areas.

       解析：未經(jīng)過無菌更衣確認的人員，不能進入A級或者B級區(qū)域。如果需要設置額外情況，制造商應該建立書面規(guī)程，來描述未經(jīng)過更衣確認人員如何進入A/B級區(qū)域，如何被管控。

       第五部分：生產(chǎn)和特定技術(shù)部分的強制性條款

       8.3 Filling of products for terminal sterilisation should be carried out in at least a grade C environment.

       解析：最終滅菌產(chǎn)品的灌裝區(qū)域應該至少設置在C級區(qū)域。

       8.22- Glass ampoules, BFS units and small volume containers (≤100 ml) closed by fusion should be subject to 100% integrity testing using validated methods.

       解析：采用融封工藝生產(chǎn)的玻璃安瓿、BFS工藝生產(chǎn)的單元包裝、小容量容器（≤100ml）應該采用驗證的方法進行100%的完整性檢測。

       8.67- All air supplied to the tunnel should pass through at least a HEPA filter and periodic tests (at least biannually) should be performed to demonstrate air filter integrity.

       解析：針對干熱滅菌，供應到隧道的所有空氣都應至少通過HEPA，并且應進行定期測試（至少每兩年一次）以證明空氣過濾器的完整性。

       從這條可以看出，用于干熱隧道烘箱的過濾器必須是HEPA，不能采用非HEPA。而且完整性測試頻率也進行詳細規(guī)定。

       8.68 When a thermal process is used as part of the depyrogenation process for any component or product contact equipment/material, validation studies should be performed to demonstrate that the process provides a suitable Fh value and results in a minimum 3 log10 reduction in endotoxin concentration.

       解析：當干熱工藝被用作任何部件或產(chǎn)品接觸設備/材料的去熱原過程時，應進行驗證研究以證明該工藝提供了合適的Fh值并導致內(nèi)毒素濃度至少降低3個log10。

       8.79 If the product cannot be sterilised in its final container, solutions or liquids should be sterilised by filtration through a sterile sterilising grade filter (with a nominal pore size of a maximum of 0.22 μm that has been appropriately validated to obtain a sterile filtrate) and subsequently aseptically filled into a previously sterilised container.

       解析：如果產(chǎn)品不能在其最終容器中滅菌，溶液或液體應通過無菌級過濾器（標稱孔徑最大為0.22 μm，經(jīng)適當驗證以獲得無菌濾液）過濾來除菌，隨后填充到已經(jīng)預先滅菌的容器中。

       第六部分：環(huán)境和工藝監(jiān)控部分的強制性條款

       9.16 For grade A, particle monitoring should be undertaken for the full duration of critical processing, including equipment assembly.

       解析：對于A級區(qū)域，應在關鍵加工的整個過程中進行懸浮粒子監(jiān)測，包括設備組裝操作。

       9.17 The grade A area should be monitored continuously (for particles ≥0.5 and ≥5 μm) and with a suitable sample flow rate (at least 28 litres (1ft3) per minute) so that all interventions, transient events and any system deterioration is captured.

       解析：應連續(xù)監(jiān)測A級區(qū)域（對于 ≥0.5um和 ≥5 μm的顆粒）并以合適的采用速度（至少28升（1ft3）每分鐘的流速），以便捕獲所有干預、瞬態(tài)事件和任何系統(tǒng)惡化信息。從這句可以看出，A級區(qū)域懸浮粒子監(jiān)控的采樣流速，需要符合上面的硬性要求。

       第七部分：質(zhì)量控制部分的強制性條款

       10.3 The bioburden assay should be performed on each batch for both aseptically filled product and terminally sterilised products and the results considered as part of the final batch review.

       解析：對無菌灌裝產(chǎn)品和最終滅菌產(chǎn)品的每批產(chǎn)品都應進行生物負荷量測定，并將結(jié)果視為最終批次審核的一部分。根據(jù)這句，應該在每批次無菌產(chǎn)品生產(chǎn)時，在合適位置取樣測試生物負荷量。

       總結(jié)

       根據(jù)上面匯總和梳理的信息可以看出，剛發(fā)布的EU GMP附錄1對于無菌藥品提出了明顯的嚴苛要求。因此說，想進軍歐盟市場的中國企業(yè)，要提前準備，才可以取得預期的成績。

       說明：本文不構(gòu)成價值判斷和投資建議。

       作者簡介：zhulikou431，高級工程師、PDA會員、ISPE會員、ECA會員、PQRI會員、資深無菌GMP專家，在無菌工藝開發(fā)和驗證、藥品研發(fā)和注冊、CTD文件撰寫和審核、法規(guī)審計、國際認證、國際注冊、質(zhì)量體系建設與維護領域，以及無菌檢驗、環(huán)境監(jiān)控等領域皆具有較深造詣。近幾年開始著力關注制藥宏觀領域趨勢分析和制藥企業(yè)并購項目的風險管理工作。