PHAGE THERAPY ACTIVE SUBSTANCES AND MEDICINAL PRODUCTS FOR HUMAN AND VETERINARY USE
5.31. 人用和兽用噬菌体治疗活性物质和药品 PHAGE THERAPY ACTIVE SUBSTANCES AND MEDICINAL PRODUCTS FOR HUMAN AND VETERINARY USE
出版这一通则是为了提供参考。
This general chapter is published for information.
它为人用和兽用噬菌体治疗活性物质和药品及其生产和控制提供了要求框架。
It offers a framework of requirements for phage therapy active substances and medicinal products for human and veterinary use and their production and control.
本章的规定不排除使用主管当局可接受的替代生产和控制方法，例如，使用无细胞系统。
The provisions of the chapter do not exclude the use of alternative production and control methods that are acceptable to the competent authority, for instance, the use of a cell-free system.
1. 定义 DEFINITION
噬菌体是感染细菌并依赖其细菌宿主进行复制的病毒。噬菌体由单链或双链
DNA或RNA组成的基因组和外层包裹的蛋白质衣壳组成。
Bacteriophages (phages) are viruses that infect bacteria and depend on their bacterial host for replication. Phages consist of a genome comprised of single or double stranded DNA or RNA, encapsulated in a protein capsid.
噬菌体治疗医药产品（PTMPs）是天然存在或转基因噬菌体的制剂，用于治疗人 类或动物细菌感染或其他医学病症。
Phage therapy medicinal products (PTMPs) are preparations of naturally occurring or genetically modified phages used to treat human or veterinary bacterial infections or other medical conditions.
PTMP可含有一种噬菌体治疗活性物质，或噬菌体混合物，或与赋形剂结合。
PTMPs可以通过多种途径给药，可赋予不同剂型。
A PTMP can contain one phage therapy active substance or a mixture of phages, combined with excipients. PTMPs can be administered by various routes and are available in different dosage forms.
2. 生产 PRODUCTION
2-1. 一般规定 GENERAL PROVISIONS
噬菌体通过在细菌宿主菌株中繁殖获得，并使用合适的方法纯化。
Phages are obtained by propagation in bacterial host strains and are purified using suitable methods.
生产过程需产生一致质量和稳定的PTMP。在相关的时间点和/或生产过程中的关 键中间阶段实施合适的过程内检测。
The production process yields a PTMP of consistent quality and stability. Appropriate inprocess testing is implemented at relevant time points and/or key intermediate stages of the process.
PTMPs的生产是基于一个合格的种子批系统，该系统使用已被验证适合的宿主
噬菌体组合。
Production of PTMPs is based on a qualified seed-lot system using a host-phage combination that has been shown suitable.
2-2. 细 菌 主 细 胞 库 和 工 作 细 胞 库 BACTERIAL MASTER CELL BANK AND WORKING CELL BANK
用于PTMP生产的细菌宿主细胞必须在菌株水平上详细表征，包括菌株来源，和 后续的操作以及用于表征菌株的测试信息。
Bacterial host cells used for PTMP production must be described in detail at the strain level. This includes information on the source of the bacterial strain, subsequent manipulations and the tests used to characterise the strain.
用于生产 PTMP 的细菌细胞来自合格的克隆来源细菌主细胞库 (MCB)，并符合 以下要求：
Bacterial cells used for PTMP production are derived from a qualified bacterial master cell bank (MCB) that is of clonal origin and complies with the following requirements:
微生物纯度：通过铺板或其他合适的方法来确定是否存在微生物污染物。
Microbial purity. The absence of microbial contaminants is determined by plating or other suitable methods.
鉴定：通过合适的表型和基因型方法，优选全基因组测序、生化或分子遗传 学测试、或质谱法来表征细菌菌株。该分型须包括抗生素敏感性谱及其染色 体和质粒的核苷酸序列。除非另有正当理由和授权，在PTMP生产中应避免 使用编码污染（如原噬菌体）或其他有害因素（如抗生素耐药性决定因素、 毒素）的菌株。
Identification. The bacterial strain is characterised by suitable phenotypic and genotypic methods, preferably whole genome sequencing, biochemical or molecular genetic tests, or mass spectrometry. This typing must include determination of its antibiotic susceptibility profile and of the nucleotide sequences of its chromosome and plasmids. The use of bacterial strains encoding contaminating (e.g. prophages) or other detrimental factors (e.g. antibiotic resistance determinants, toxins) is avoided in the production of PTMPs, unless otherwise justified and authorised.
细胞活力：活细胞的数量由平板计数或其他合适的活细胞计数方法确定。
Viability. The number of viable cells is determined by a plate count or other suitable viable cell count methods.
噬菌体敏感性：菌株对噬菌体治疗活性物质的敏感性用噬菌斑试验或其他合 适的方法来证明。
Phage sensitivity. The susceptibility of the strain to the phage therapy active substance is demonstrated using a plaque assay or other suitable methods.
无不利噬菌体：证实没有可能对PTMPs不利的噬菌体（如溶原性噬菌体）。
Absence of detrimental phages. The absence of phages that could be detrimental to the production of PTMPs (e.g. lysogenic phages) is confirmed.
用于生产的工作细胞库是主细胞库的克隆衍生物，并且符合活力、微生物纯度和噬菌体敏感性的要求。
The working cell bank to be used for production is a clonal derivative of the MCB and complies with the requirements for viability, microbial purity and phage sensitivity.
2-3. 用于生产PTMPs的噬菌体 PHAGES USED FOR PRODUCTION OF PTMPs
PTMP生产中使用的噬菌体须详细表征。需提供有关噬菌体来源、核苷酸序列和 易感细菌种类的信息。除非另有正当理由和授权，应避免使用编码已知或潜在有 害遗传因素的噬菌体，如抗生素耐药性决定因素、毒素、溶原性模块或其他元素。
Phages used in PTMP production must be characterised in detail. Information on the phage source, nucleotide sequence and susceptible bacterial species is to be provided. Phages encoding known or potential detrimental genetic factors, e.g. antibiotic resistance determinants, toxins, lysogeny modules or other elements, are avoided unless otherwise justified and authorised.
只有符合以下要求的主噬菌体库（MPBs）才能用于生产：
Only master phage banks (MPBs) that comply with the following requirements may be used for production:
鉴定：噬菌体通过包括噬菌斑形态和全基因组测序等方法在内的合适的表型 和基因型方法进行鉴定。对于基因或化学修饰的噬菌体，修饰及其效果必须 具体描述和鉴定。
Identification. The phage is identified by suitable phenotypic and genotypic methods, including plaque morphology and whole genome sequencing. For genetically or chemically modified phages, the modifications must be described and their effects characterised.
噬菌体纯度：通过反复的噬菌斑纯化来纯化噬菌体，直到获得具有均一噬菌 斑形态的噬菌体裂解液，其中包含一株噬菌体的克隆。
Phage purity. The phage is purified by repeated rounds of plaque purification until a phage lysate with one plaque morphotype is obtained, containing one phage clone.
效价：通过噬菌斑试验或其他合适的方法确定感染性噬菌体的滴度。
Potency. The infectious phage titre is determined by a plaque assay or other suitable method.
无菌（2.6.1）：需符合无菌检验。
用于生产的工作噬菌体库是主噬菌体库的克隆衍生物，需符合噬菌体纯度、 无菌和效价要求。
Sterility (2.6.1). It complies with the test for sterility. The working phage bank to be used for production is a clonal derivative of the MPB and complies with the requirements for phage purity, sterility and potency.
2-4. 生产和纯化 PRODUCTION AND PURIFICATION
PTMPs的生产基于种子批系统，应严格避免不同噬菌体和细菌宿主菌株之间的交 叉污染。
Production of PTMPs is based on a seed-lot system, in which cross-contamination between different phage and bacterial host strains is strictly avoided.
采用药级原料。生物来源的原材料也需符合通则5.2.12的要求。用于生产细胞基 因治疗药品的生物原料。
Raw materials of pharmaceutical grade are used. Raw materials of biological origin also comply with the requirements of general chapter 5.2.12. Raw materials of biological origin for the
production of cell-based gene therapy medicinal products.
噬菌体通过合适的技术纯化。
Phages are purified by suitable techniques.
同一噬菌体克隆的几个单次收获物可能在纯化过程之前合并。
Several single harvests of the same phage clone may be pooled before the purification process.
只有符合以下要求的纯化后收获物才能用于制备成品批：
Only a purified harvest that complies with the following requirements may be used in the preparation of the final lot:
鉴定：使用相关的基因型和表型标记物来确认噬菌体的身份。
Identification. Identity of the phage is confirmed using relevant genotypic and phenotypic markers.
效价：感染性噬菌体的滴度通过空斑试验或其他合适的方法确定。
Potency. The infectious phage titre is determined by a plaque assay or other suitable method.
微生物学检查（2.6.12）：符合既定的规范。
Microbiological examination (2.6.12). It complies with the established specification.
残留试剂：在风险分析的基础上，对纯化的收获物进行生产过程中使用的试 剂残留和安全风险的测试。
Residual reagents. Based on risk analysis, the test for residues of reagents used during production and that pose safety concerns is carried out on the purified harvest.
宿主细胞杂质和污染物：来自宿主细胞的污染物和其他潜在有毒物质（如内 毒素和外毒素、宿主细胞蛋白质、宿主细胞DNA）不存在或在特定制剂批准 的限度内。
Host-cell impurities and contaminants. Contaminants and other potentially toxic substances derived from the host cells (e.g. endo- and exotoxins, host-cell proteins, hostcell DNA) are absent or within the limits approved for the particular preparation.
2-5. 成品批 FINAL LOT
成品批可以配制成几种不同的剂型：固体（如片剂）、液体（如注射或口服）或 冻干制剂。
The final lot may be formulated as several different dosage forms: solid (e.g. tablets), liquid (e.g. injectable or for oral administration) or freeze-dried preparations.
成品批符合以下要求：
A final lot complies with the following requirements:
外观：符合既定规范。
Appearance. It complies with the established specification.
鉴定：使用相关的基因型和表型标记物来确认噬菌体的身份。 Identification. The identity of each phage is verified using relevant genotypic and phenotypic markers.
效价： 每种噬菌体的感染性滴度通过噬菌斑测定或其他合适的方法测定， 以PFU/mL或PFU/mg表示，并且在特定制剂批准的限度内。
Potency. The infectious phage titre of each phage, expressed in PFU/mL or PFU/mg, is determined by a plaque assay or other suitable method and is within the limits approved for the particular preparation.
无菌（2.6.1）：对于无菌PTMPs，符合无菌检验。
Sterility (2.6.1). Sterile PTMPs, comply with the test for sterility.
微生物学质量：对于非无菌PTMPs，通则5.1.4章提供了建议。药用非无菌制 剂和物质的微生物学质量。 Microbiological quality. For non-sterile PTMPs, recommendations are provided in general chapter 5.1.4. Microbiological quality of non-sterile pharmaceutical preparations and substances for pharmaceutical use.
热原性 [1] (5.1.13)：特定的制剂，如果适用的话应符合合适的热原性检测。
Pyrogenicity(1) (5.1.13). The particular preparation, if applicable, complies with a suitable test for pyrogenicity.
冻干PTMPs还需符合以下附加要求：
Freeze-dried PTMPs comply with the following additional requirement:
水含量 (2.5.12 or 2.5.32)：符合特定产品批准的限制。 Water content (2.5.12 or 2.5.32). It complies with the limit approved for the particular product.
液态PTMPs还需符合以下附加要求： Liquid PTMPs comply with the following additional requirement:
pH (2.2.3)：符合特定产品批准的限制。 pH (2.2.3). It complies with the limit approved for the particular product.
2-6. 适应性产品 ADAPTED PRODUCT
噬菌体适应是指PTMP可以通过定向进化来增加其对个体患者临床分离细菌株 的效力的过程。噬菌体适应所用单个噬菌体或噬菌体组合符合第2-3节规定。成 品批的适应性PTMP的效价使用目标临床分离菌株来检测。除非另有正当理由和 授权，成品批应符合第2-5节外观、无菌（或微生物污染）和热原性的规定。
Phage adaptation is the process by which a PTMP can be directed to evolve in order to increase its potency against a clinical isolate from an individual patient. Phage adaptation starts with a phage or mixture of phages that comply with the provisions of section 2-3. The potency of the final lot of the adapted PTMP is determined against the target clinical isolate. The final lot complies with the provisions for appearance, sterility (or microbial contamination) and pyrogenicity of section 2-5, unless otherwise justified and authorised.
3. 标签 LABELLING
适用相关超国家和国家法规中概述的标签要求。 The labelling requirements outlined in relevant supranational and national regulations apply.
[1] 药典35.1纳入一章新通则——热原性（5.1.13），提供了热原检测方法选择的 指导。
[1] A new general chapter, Pyrogenicity (5.1.13), which provides guidance on the choice of pyrogen testing approach, is currently published in Pharmeuropa 35.1.
© Pharmeuropa 35.2
Reference: PA/PH/Exp. BACT/T (22) 1 ANP
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