Ocriplasmin

Entered strategic partnership with Otsuka Pharmaceutical Co., Ltd. for the development and commercialization of 4D-150 in the APAC region; to receive $85 million in upfront cash and expects to receive at least $50 million from cost sharing Announced positive long-term safety and efficacy data with 1.5 to 2 years of follow-up from the Phase 1/2 PRISM clinical trial in wet AMD Completed equity offering providing net proceeds of ~$93 million Announced up to $11 million equity investment from the Cystic Fibrosis Foundation to accelerate development of 4D-710 for cystic fibrosis into Phase 2 $372 million in cash, cash equivalents and marketable securities as of September 30, 2025, combined with upfront and expected cost sharing from Otsuka partnership and ~$93 million in net proceeds from equity offering, expected to fund currently planned operations into second half of 2028 EMERYVILLE, Calif., Nov. 10, 2025 (GLOBE NEWSWIRE) -- 4D Molecular Therapeutics (Nasdaq: FDMT, 4DMT or the Company), a leading late-stage biotechnology company advancing durable and disease-targeted therapeutics with potential to transform treatment paradigms and provide unprecedented benefits to patients, today reported Q3 2025 financial results, provided operational highlights and outlined expected upcoming milestones.  “In the third quarter, we made meaningful progress building upon and validating our business strategy focusing on 4D-150 and 4D-710,” said David Kirn, M.D., Co-founder and Chief Executive Officer of 4DMT. “Our recently announced partnership with Otsuka, new data on 4D-150 in wet age-related macular degeneration, equity investment from the Cystic Fibrosis Foundation for advancement of 4D-710 and strengthened balance sheet position us well, with cash runway into 2H 2028, beyond primary readout for our two ongoing 4D-150 4FRONT Phase 3 trials in wet AMD, and provides for indication expansion in diabetic macular edema.” Recent Corporate Highlights Announced Exclusive License Agreement with Otsuka Pharmaceutical Co., Ltd. for Development and Commercialization of 4D-150 in Asia-Pacific Region: 4DMT to receive $85 million upfront cash payment and expects to receive at least $50 million of cost sharing from Otsuka over the next three years for development activities supporting global registration    The Company is eligible for up to $336 million in potential regulatory and commercial milestones and tiered double-digit royalties 4DMT retains full development and commercialization rights for 4D-150 outside the APAC region, including the U.S., Europe and Latin America Completed an Equity Offering with Approximately $93 Million of Net Proceeds: Proceeds to support planned operations into second half of 2028, which include planned 4D-150 Phase 3 clinical trial in diabetic macular edema (DME), and provide more than 12 months of expected cash runway beyond the expected 4FRONT-1 topline data Strengthened Leadership Team to Drive 4D-150 Program Execution Julie Clark, M.D., was promoted to Chief Medical Officer, bringing over 20 years of experience in retina and global clinical development, with a proven track record across 10 BLA-enabling studies, including six Phase 3 trials, with leadership roles in development and approvals of EYLEA®, JETREA®, BEOVU® and IZERVAY®  Liansheng Zhu, Ph.D., joined as SVP, Biometrics and Data Quality, bringing nearly two decades of global experience in late-stage and post-approval clinical development. His expertise includes biostatistics, statistical programming, data science and Health Economics and Outcomes Research. Dr. Zhu has led or contributed to multiple pivotal clinical trials and regulatory submissions resulting in efficient and successful U.S. Food and Drug Administration (FDA) and European Medicines Agency (EMA) approvals, including for LUCENTIS®, BEOVU® and IZERVAY®  Created Retina Leadership Advisory Board: Glenn P. Sblendorio – Former President and CEO of Iveric BioCal Roberts, M.D. – Former CEO of Lighthouse Guild, Former Chief Medical Officer of Bausch + Lomb, Clinical Professor of Ophthalmology at Weill Cornell Medical Center Wiley A. Chambers, M.D. – Former Director of the Division of Ophthalmology, Center for Drug Evaluation and Research at the FDA Recent Highlights and Expected Milestones for 4D-150 4D-150 for Wet AMD: 4FRONT Global Phase 3 Program: 4FRONT-1, North American Clinical Trial: Enrollment rate exceeds initial expectations, with over 200 patients randomized to date, and is on track to complete enrollment in Q1 2026, with 52-week topline data expected in H1 2027 4FRONT-2, Global Clinical Trial: Enrollment remains on track to be completed in H2 2026, with 52-week topline data expected in H2 2027  PRISM Phase 1/2 Clinical Trial: Announced positive long-term interim results (data cutoff: August 22, 2025)  4D-150 demonstrated consistent and durable benefit across all three patient cohorts as evidenced by maintenance of visual acuity, control of retinal anatomy and reduction of treatment burden at all time points with 1.5-2 years of follow-up Consistent dose response in favor of Phase 3 dose (3E10 vg/eye) continues to be demonstrated across all wet AMD cohorts studied 4D-150 continues to be well tolerated with no new safety or intraocular inflammation findings with up to 3.5 years of follow-up   4D-150 for DME: SPECTRA Clinical Trial: Presented positive 60-week results (data cutoff: May 3, 2025)   4D-150 continues to be well tolerated across all patients and dose levels, with no intraocular inflammation observed at any time point or dose levelFollowing the three loading doses of aflibercept, 3E10 vg/eye, the Phase 3 dose demonstrated strong signals of clinical activity, with sustained gain of BCVA of +9.7 letters and reduction of CST of -174 µm from baseline Supplemental injections: Phase 3 dose achieved 78% reduction in injection burden vs. projected on-label aflibercept 2mg Q8W Dose response observed for the Phase 3 dose vs. lower doses (58% fewer injections) Recent Highlights and Expected Milestones in 4D-710 Program 4D-710 for Cystic Fibrosis (CF) Lung Disease: Secured an equity investment from the CF Foundation of up to $11 million in two tranches, with the first tranche of $7.5 million received in October 2025 The funding will support:  Phase 1 Redosing and Phase 2 Cohort in AEROW clinical trial Phase 3 Readiness Interim safety and efficacy data from AEROW Phase 1 clinical trial expected by year-end 2025  Q3 2025 Financial Results Cash Position: Cash, cash equivalents and marketable securities were $372 million as of September 30, 2025, as compared to $505 million as of December 31, 2024. The net decrease in cash was primarily a result of cash used in operations. Based on our current operating plan, we estimate that our existing cash, cash equivalents and marketable securities, and expected payments under our collaboration agreement with Otsuka, together with the net proceeds from the November 2025 equity offering, will be sufficient to fund our operating expenses and capital expenditure requirements at least into the second half of 2028. R&D Expenses: Research and development expenses were $49.4 million for the third quarter of 2025, as compared to $38.5 million for the third quarter of 2024. This increase was primarily driven by the Phase 3 clinical trials of 4D-150 in wet AMD. G&A Expenses: General and administrative expenses were $11.8 million for the third quarter of 2025, as compared to $12.7 million for the third quarter of 2024. The decrease was primarily due to decreased headcount of general and administrative personnel. Net Loss: Net loss was $56.9 million for the third quarter of 2025, as compared to net loss of $43.8 million for the third quarter of 2024. About 4DMT 4DMT is a leading late-stage biotechnology company advancing durable and disease-targeted therapeutics with potential to transform treatment paradigms and provide unprecedented benefits to patients. The Company’s lead product candidate 4D-150 is designed to be a backbone therapy forming the foundation of treatment of blinding retinal vascular diseases by providing multi-year sustained delivery of anti-VEGF (aflibercept and anti-VEGF-C) with a single, safe, intravitreal injection, which substantially reduces the treatment burden associated with current bolus injections. The Company’s lead indication for 4D-150 is wet age-related macular degeneration, which is currently in Phase 3 development, and second indication is diabetic macular edema. The Company’s second product candidate is 4D-710, which is the first known genetic medicine to demonstrate successful delivery and expression of the CFTR transgene in the lungs of people with cystic fibrosis after aerosol delivery. 4D Molecular Therapeutics™, 4DMT™, Therapeutic Vector Evolution™, and the 4DMT logo are trademarks of 4DMT.   All of the Company’s product candidates are in clinical or preclinical development and have not yet been approved for marketing by the U.S. Food and Drug Administration or any other regulatory authority. No representation is made as to the safety or effectiveness of the Company’s product candidates for the therapeutic uses for which they are being studied.  Learn more at www.4DMT.com and follow us on LinkedIn. Forward-Looking Statements: This press release contains forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995, as amended, including, without limitation, implied and express statements regarding the therapeutic potential and clinical benefits of, as well as the plans, announcements and related timing for the clinical development of our product candidates, the potential benefits of the strategic partnership with Otsuka, the amount of any potential cost sharing or milestone payments pursuant to the Company’s agreement with Otsuka, the Company’s use of proceeds, the potential benefits of the investment from and collaboration with the CF Foundation, the potential additional second tranche funding from the CF Foundation and statements regarding our financial performance, results of operations and anticipated cash runway. The words "may," “might,” "will," "could," "would," "should," "expect," "plan," "anticipate," "intend," "believe," “expect,” "estimate," “seek,” "predict," “future,” "project," "potential," "continue," "target" and similar words or expressions are intended to identify forward-looking statements, although not all forward-looking statements contain these identifying words. Any forward-looking statements in this press release are based on management's current expectations and beliefs and are subject to a number of risks, uncertainties and important factors that may cause actual events or results to differ materially from those expressed or implied by any forward-looking statements contained in this press release, including risks and uncertainties that are described in greater detail in the section entitled "Risk Factors" in 4D Molecular Therapeutics’ most recent Quarterly Report on Form 10-Q filed on or about the date hereof, as well as any subsequent filings with the Securities and Exchange Commission. In addition, any forward-looking statements represent 4D Molecular Therapeutics' views only as of today and should not be relied upon as representing its views as of any subsequent date. 4D Molecular Therapeutics explicitly disclaims any obligation to update any forward-looking statements. No representations or warranties (expressed or implied) are made about the accuracy of any such forward-looking statements.  4D Molecular Therapeutics, Inc.Statements of Operations(Unaudited)(in thousands, except share and per share amounts) Three months endedSeptember 30, Nine months endedSeptember 30, 2025 2024 2025 2024 Revenue: Collaboration and license revenue $90 $3 $119 $36 Operating expenses: Research and development 49,438 38,484 138,088 98,212 General and administrative 11,837 12,651 36,293 33,548 Total operating expenses 61,275 51,135 174,381 131,760 Loss from operations (61,185) (51,132) (174,262) (131,724)Other income, net 4,309 7,289 14,756 20,527 Net loss $(56,876) $(43,843) $(159,506) $(111,197)Net loss per share, basic and diluted $(1.01) $(0.79) $(2.85) $(2.08)Weighted-average shares outstanding used in computing net loss per share, basic and diluted 56,126,330 55,554,476 55,933,890 53,377,712 4D Molecular Therapeutics, Inc.Balance Sheet Data(Unaudited)(in thousands) September 30,2025 December 31,2024 Cash, cash equivalents and marketable securities $372,228 $505,460 Total assets 423,982 560,384 Total liabilities 54,999 49,778 Accumulated deficit (735,701) (576,195)Total stockholders’ equity 368,983 510,606 Contacts: Media: Jenn Gordondna CommunicationsMedia@4DMT.com Investors: Julian PeiHead of Investor Relations and Strategic FinanceInvestor.Relations@4DMT.com

👆 关注我，一起学习最新文献、工艺技术和法规知识今天分享的是《The Challenge of CMC Regulatory Compliance for Biopharmaceuticals》这本书的第十章，生物制品工艺相关杂质的复杂性。药物生产过程中，工艺相关杂质的残留不可避免，因此必须对其潜在的临床安全性风险进行全面评估，以确保患者用药安全。与化学药物相比，生物制品（无论是重组蛋白、单克隆抗体、病毒载体、经基因改造的患者细胞，还是mRNA非病毒载体）由于采用生物生产工艺，具有高度复杂的工艺相关杂质安全性特征。本章将针对生物制品生产工艺中产生的众多工艺相关杂质，基于风险的控制策略，探讨满足最低CMC监管合规性要求的风险方法。供大家学习参考。整个生物药生产工艺流程——从起始物料→原液上游生产→原液下游纯化→处方→制剂灌装/密封——都会影响最终成品中工艺相关残留杂质的种类和含量。例如，对于重组蛋白而言，细胞培养生产运行时间的延长不仅会影响生物蛋白的总体产量，还会因生产细胞随培养时间延长而裂解，导致收获液中的宿主细胞蛋白（HCP）水平升高。HCP含量的增加对下游纯化工艺提出了更高要求，若HCP含量过高可能导致纯化能力超载。因此，有效控制工艺相关杂质需要从起始物料到最终成品的整个生物工艺流程进行基于风险的全面评估与研究。1.  何为工艺相关杂质社会学家威廉·布鲁斯·卡梅伦（William Bruce Cameron）有一句格言：“能计算的不一定重要，重要的不一定能计算。”若将其应用于工艺相关杂质，似乎存在无穷无尽的杂质清单（“任何进入生产体系但未成为最终药品的物质均可视为杂质”）。此外，所有工艺相关杂质对患者的安全风险等级并非完全相同，因此需要不同程度的残留控制。首先介绍三个易于混淆的术语：工艺相关杂质（Process-Related Impurities）：源自生产工艺的杂质。污染物（Contaminants）：非工艺设计引入的意外物质（如微生物污染）。产品相关杂质（Product-Related Impurities）：目标产物的分子变体，但其活性、有效性或安全性不及目标产物。本章聚焦工艺相关杂质。工艺相关杂质因其对患者的安全性有潜在影响而被作为产品关键质量属性（CQAs）的重要组成部分。化学药的工艺相关杂质主要为有机杂质和无机杂质，而生物药的工艺相关杂质种类更为复杂，包括：宿主来源杂质（如宿主细胞蛋白HCP、DNA等）细胞培养过程杂质（如培养基成分等）下游工艺杂质（如层析柱浸出物等）根据ICH指南（Q3A、Q3C、Q6B），生物药的CQAs需综合考虑工艺相关杂质和污染物（如病毒污染）。2.  基于生产工艺类型的工艺相关杂质不同类型的生物药生产工艺均需对其各步骤进行风险评估，以确定其对工艺相关杂质生成/清除的贡献程度（如下图所示的石川图/鱼骨图）。目前已有超过200种获批上市的重组蛋白和单克隆抗体，关于此类生物药生产工艺中可能涉及的工艺相关杂质类型已有较充分的认知：依据ICH Q6B Specifications – Test Procedures and Acceptance Criteria for Biotechnological/Biological Products (March 1999)：“ 工艺相关杂质：这些杂质源自生产工艺，可分为三大类：宿主来源杂质、细胞培养过程杂质和下游工艺来源杂质。(a) 宿主细胞来源杂质包括但不限于宿主细胞来源的蛋白质、核酸（宿主细胞基因组、载体DNA或总DNA）。对于宿主细胞蛋白（HCP），通常采用高灵敏度的免疫分析法（如ELISA）来检测广谱的蛋白质杂质，用于免疫分析法检测的多克隆抗体是通过用不含目标产物编码基因的宿主细胞制备。宿主细胞DNA的水平可通过对产品进行直接分析（例如杂交技术）来检测。清除研究（可能包括实验室规模的加标实验）可用于证明宿主细胞来源的杂质（如核酸和宿主细胞蛋白）的去除能力，从而可能免除为这些杂质建立质量标准的需要。(b) 细胞培养过程杂质包括但不限于：诱导剂、抗生素、血清及其他培养基成分。(c) 下游工艺来源杂质包括但不限于：酶（如蛋白酶）、化学试剂（如溴化氰、胍盐、氧化/还原剂）；无机盐（如重金属、砷、非金属离子）；溶剂、载体、配体（如单抗）；层析柱浸出物“下表总结了重组蛋白/单抗生产各步骤中潜在工艺相关杂质的来源：2.1  DS上游生产上游生产工艺是蛋白类生物药中工艺相关杂质的主要来源。潜在的上游生产杂质来源于细胞培养基中的多种原材料。培养基需提供细胞生长所需的各种氨基酸、维生素、糖类和无机盐。此外，还可能包含以下成分：血清（如胎牛血清，FBS）、生长因子（如胰岛素样生长因子，IGF-1）、脂质（如胆固醇）、选择性试剂（如甲氨蝶呤）、抗生素（如庆大霉素）、消泡剂（如Pluronic F-68）等。这些成分均可能成为工艺相关杂质。此外，宿主细胞本身也会产生潜在的杂质（如宿主细胞DNA和宿主细胞蛋白）。胎牛血清（FBS）可以刺激动物和人类细胞生长和增殖，并有助于保持生产细胞在一段时间内的正常和健康状态。胎牛血清含有牛血清白蛋白（BSA）、免疫球蛋白（IgG）以及丰富的其他蛋白质混合物。然而，胎牛血清不仅带来了朊病毒的风险，而且当作为残留杂质存在时，还会引发免疫原性的担忧。胰岛素和胰岛素样生长因子-1（IGF-1）是广泛使用的生长因子，用于延缓哺乳动物细胞培养中的细胞凋亡，从而提高生产效率。这些生长因子通过增加特定的葡萄糖消耗率来降低细胞内活性氧水平，从而在无血清培养中抑制细胞死亡。然而，生物药产品中残留的生长因子可能会因其激素活性而引起关注。基因扩增系统用于提高重组蛋白或单克隆抗体的产量。在哺乳动物细胞中，通常采用以下两种系统维持筛选压力：二氢叶酸还原酶（DHFR）系统：通过甲氨蝶呤（MTX）抗性筛选（常用于CHO细胞）；谷氨酰胺合成酶（GS）系统：通过蛋氨酸亚砜亚胺（MSX）抗性筛选（适用于CHO和NS0细胞）。MTX虽然在医学上用于治疗某些自身免疫性疾病和癌症，但可能会导致多种器官毒性；而MSX是一种潜在的神经毒素。因此，尽管MTX或MSX仅在早期种子培养中使用，但它们可能会作为残留物存在于产品中，从而引起毒性方面的担忧。抗生素在生物制品的细胞培养生产中被用作遗传筛选剂，或者用于在细胞培养生产过程中预防细菌和支原体污染。对于重组细菌细胞系（例如大肠杆菌），四环素和卡那霉素被用作重组蛋白生产过程中的遗传筛选剂。重组细胞系中的质粒除了含有蛋白基因外，还含有赋予抗生素抗性的基因片段。如果重组细胞丢失了质粒，蛋白表达就会停止，其抗生素抗性也会随之消失。此时，抗生素会关闭对细胞生存至关重要的细胞蛋白合成。通过这种方式，培养基中的营养物质得以保留，供那些继续生产重组蛋白的细胞使用。对于哺乳动物细胞系（如CHO），在重组蛋白或单克隆抗体的生产过程中，有时会在细胞培养基中添加庆大霉素等抗生素，以预防微生物或支原体污染。需要注意的是，由于存在严重的过敏反应风险（如果药物中残留有这类抗生素），β-内酰胺类抗生素（例如青霉素）被限制用于细胞培养生产。当在重组蛋白或单克隆抗体的细胞培养生产中添加抗生素时，必须将其去除，如下表所示。此外，对于生产过程中使用任何抗生素，都需要有合理的理由，例如，对于在大肠杆菌生产的重组蛋白Palynziq（pegvaliase-pqpz）中使用卡那霉素的抗生素，其上市后承诺就说明了这一点：依据FDA Drugs – Search Drugs@FDA – FDA Approved Drug Products: Palynziq (Pegvaliase-pqpz): Approval History, Letters, Reviews and Related Documents – FDA Approval Letter (May 24, 2018)：“ 开展一项研究，以评估在pegvaliase发酵工艺中去除卡那霉素对产品的影响。如果数据支持去除卡那霉素，那么提交一份从pegvaliase生产工艺中去除卡那霉素的研究计划。”还需注意的是，宿主细胞壁成分（如多糖类物质）也可能成为工艺相关杂质。在毕赤酵母（Pichia pastoris）生产的重组蛋白药物Jetrea（ocriplasmin）的上市批准中，FDA要求进行上市后承诺研究，以评估药物成分中酵母细胞壁成分的潜在影响：依据FDA Drugs – Search Drugs@FDA – FDA Approved Drug Products: Jetrea (Ocriplasmin): Approval History, Letters, Reviews and Related Documents – Chemistry Reviews – Quality Team Leader’s Executive Summary (September 13, 2012)“ 需评估药物成分中是否含有酵母细胞壁成分（如几丁质、甘露聚糖、β-葡聚糖），评估这些产品相关杂质对ocriplasmin质量、安全性和有效性的潜在风险，并提出适当的控制策略。”2.2  DS下游纯化下游纯化工艺的设计旨在应对上游生产过程中产生的工艺相关杂质，无论其来源于细胞培养、转基因动物还是转基因植物。然而，下游纯化工艺本身也可能引入新的工艺相关杂质。潜在下游工艺杂质包括：酶（如蛋白酶、核酸酶）、化学试剂（如溴化氰、胍盐、氧化/还原剂）、浸出的金属元素（如金属螯合层析树脂释放的金属离子）、浸出的配体（如Protein A亲和层析柱释放的配基）等。这些下游工艺相关杂质均需评估。Protein A亲和层析是单克隆抗体（mAb）纯化的重要步骤。Protein A是一种源自金黄色葡萄球菌的细胞壁蛋白，可特异性结合IgG的Fc结构域。然而，亲和层析树脂中浸出的Protein A可能会进入产品并引发免疫反应。由于Protein A层析柱成本高昂，生产商通常希望重复使用数百次，因此需研究树脂在使用周期前后的浸出量：依据FDA Guidance for Industry: Monoclonal Antibodies Used as Reagents in Drug Manufacturing (March 2001)：“ 在制定原液质量标准时，需考虑层析介质浸出的mAb或杂质。层析柱浸出物的量在层析柱生命周期内并非恒定，其受多种因素影响（如储存时间、再生/消毒步骤的溶液、柱操作参数等）。检测方法包括：在上样前检测缓冲液流出液、中间产物的过程检测，或终产品检测。若后续生产步骤可将浸出物降至安全水平，可豁免常规检测。”此外，Protein A填料供应商通常会提供ELISA检测试剂盒，用于定量检测纯化产品中的Protein A残留。层析纯化中可能使用有机溶剂（如异丙醇），此时需遵循ICH Q3C(R5)《残留溶剂指南》。因残留溶剂无治疗作用，应尽可能去除以满足质量标准、GMP或其他质量要求。2.3  DS的偶联对于大多数生物制品而言，元素杂质残留对患者的安全风险较低，但抗体药物偶联物（ADCs）例外，因其偶联的化学药物可能引入元素杂质：依据ICH Q3D(R2) Guideline for Elemental Impurities (April 2022)：“ 对生物技术产品而言，DS阶段的元素杂质引发安全性担忧的风险较低，主要原因包括：a) 生物制品生产过程中通常不使用元素作为催化剂或试剂；b) 细胞培养过程中添加的微量元素（如金属离子）不会蓄积，且经后续工艺被稀释/清除；c) 生物药纯化步骤（如层析、超滤/透析）可有效去除源自培养基或生产设备的元素杂质至可忽略水平。因此，生物药原液通常无需特别控制元素杂质。但对于含合成结构的产品（如ADCs），需对小分子组分（如化学连接子/毒素）中的元素杂质进行适当控制。”2.4  DP处方制剂处方中包含多种辅料，可能会向产品中引入工艺相关杂质。为确保辅料质量，需关注以下方面：采用符合药典标准（如USP或Ph. Eur.）的原料、建立供应商质量/供应链协议，以及防止降解。以聚山梨酯为例说明，聚山梨酯是聚氧乙烯山梨醇酐脂肪酸酯的混合物，其脂肪酸组成具有异质性。该辅料在商业化重组蛋白和单抗制剂中能有效防止蛋白聚集。然而，多种因素可能影响其质量：原料杂质、储存温度、其他辅料相互作用、光照等。聚山梨酯的降解主要通过氧化和水解两种途径。宿主细胞来源的酶（如大肠杆菌、CHO细胞或酵母中的酯酶）可快速水解脂肪酸链，导致含聚山梨酯的制剂中形成亚可见或可见颗粒。2.5  制剂灌装/密封过程中产生的杂质制剂的灌装/密封工艺可能向生物制品中引入工艺相关杂质，例如残留过氧化氢和过量硅油。无菌处理对于生物制品的灌装过程至关重要。气态过氧化氢（VPHP）作为一种隔离器和洁净室的生物去污剂，已成为灌装操作的标准方法。然而，残留的过氧化氢可能会氧化重组蛋白和单克隆抗体（尤其是蛋氨酸、色氨酸和半胱氨酸这些氨基酸）。生物药溶液在灌装线上的硅胶管中流动，以及在无菌环境中开放的玻璃小瓶，被认为是药物产品吸收残留过氧化氢的重要来源。硅油需用于涂覆玻璃瓶的橡胶塞或预充式注射器的橡胶活塞，以确保密封性或推动药液。然而，过量硅油可能导致产品溶液中形成硅油液滴，从而促进蛋白聚集。生物制品与包材接触的表面可能产生浸出物。对于制剂中的元素杂质残留，需进行基于风险的评估：依据ICH Q3D(R2) Guideline for Elemental Impurities (April 2022)：“ 生物技术药物需考虑制剂生产中的潜在元素杂质来源（如辅料）及环境因素。由于这些杂质通常在制剂生产后期引入且无后续清除步骤，其风险需综合评估，包括辅料类型、工艺条件、环境控制水平（如无菌生产区、纯化水使用）及给药频率等。”3.  最低限度的CMC法规符合性监管机构接受生物制品生产采用基于风险的最低限度CMC法规符合性方法。针对重组蛋白和单克隆抗体的工艺相关杂质控制，在早期临床开发阶段（包括首次人体试验，FIH）提供以下指导：依据EMA Guideline on the Requirements for Quality Documentation Concerning Biological Investigational Medicinal Products in Clinical Trials (January 2022)：--S.3.2 杂质“ 需明确工艺相关杂质（如宿主细胞蛋白、宿主细胞DNA、培养基残留、层析柱浸出物）和产品相关杂质（如前体、裂解形式、降解产物、聚集体）。应提供杂质的定量信息，包括最高临床剂量下的最大残留量。对于某些工艺相关杂质（如消泡剂），可基于清除率估算进行合理性论证。若仅提供定性数据，需说明理由。”--S.4.1 质量标准“应根据安全性考虑设定杂质上限。”--P.5.1 质量标准“需基于安全性设定杂质上限，并在后续开发中可以重新评估和调整。对于未纳入原液标准的杂质，制剂的质量标准仍需根据安全性评估设定上限。”随着生物制品从早期临床开发进入后期临床阶段，对工艺相关杂质的理解与控制将逐步深化。在提交蛋白类生物药上市申请时，监管机构要求：1）明确工艺相关杂质的控制策略；2）合理论证杂质残留限值的设定依据。ICH M4Q(R1) The Common Technical Document for the Registration of Pharmaceuticals for Human Use: Quality – Quality Overall Summary of Module 2, Module 3 Quality (September 2002)指出：--杂质“ 除评估原液与制剂的纯度（可能包含目标产物及多种产品相关物质）外，生产企业还需评估可能存在的杂质。杂质可分为工艺相关或产品相关杂质，其结构可能已知、部分表征或未知。若可获取足量杂质样品，应尽可能表征其性质并评估生物活性。工艺相关杂质包括源自生产工艺的成分（如宿主细胞蛋白、宿主DNA、培养基成分、下游工艺残留）。此外，杂质接受标准应基于临床前/临床研究批次及工艺一致性批次的数据设定，需为单个或总体杂质设定适当限值。某些情况下，特定杂质的检测可豁免：若能通过研究证明其已被有效控制或清除至安全水平，则无需纳入质量标准。此验证需依据地方法规在商业化规模完成。申报时数据可能有限，因此可在上市后承诺继续按法规实施。”FDA对生物制品上市许可申请（BLA）的要求：依据FDA Guidance for Industry: For the Submission of Chemistry, Manufacturing, and Controls Information for a Therapeutic Recombinant DNA-Derived Product or a Monoclonal Antibody for In Vivo Use (August 1996)：“ 需提供杂质谱的讨论及支持性分析数据，包括产品相关变体（如裂解形式、聚集体、脱酰胺化、氧化形式等）和非产品相关杂质（如工艺试剂、细胞培养基成分等）。”EMA对上市许可申请（MAA）的要求：依据EMA Guideline on Process Validation for the Manufacture of Biotechnology-Derived Active Substances and Data to be Provided in the Regulatory Submission (April 2016)：“ 应全面评估纯化工艺去除产品和工艺相关杂质（如非目标变体、HCP、核酸、培养基成分、病毒及蛋白修饰试剂等）的能力，确保其降至可接受水平。通常需建立适用于杂质检测的分析方法，并评估各纯化步骤的生产或清除能力，从而设定合理的接受标准。对于某些工艺相关杂质（如HCP、DNA、抗生素等），需通过缩小规模加标实验验证各纯化步骤的清除能力。对于定义为高清除率的步骤，应在最差条件和/或非标准条件（如工艺停留时间、加标挑战）下验证工艺稳健性。对于部分成分（如低分子量培养基组分），可采用基于风险的方法证明其无免疫原性或毒性风险。”以下是已上市单链Fv抗体片段Beovu（brolucizumab）的工艺相关杂质分析案例：依据EMA Medicines: European Public Assessment Report: Beovu (Brolucizumab) EPAR (December 2019)：“ 通过验证实验评估生产工艺对工艺相关杂质（HCP、宿主细胞DNA、培养基及缓冲液成分）的清除能力，监测各工艺步骤的样品。针对HCP和宿主DNA进行了加标实验。结果显示，现有工艺能持续有效地将杂质降至极低水平，多数工艺相关杂质低于定量限（LOQ）且未超出安全性阈值（SCT，1.5 μg/天）。对生产过程中接触产品、中间液、缓冲液、培养基的包材进行了浸出物研究（包括毒理学评估）。基于综合风险评估及模拟使用条件下的浸出物研究得出结论，brolucizumab生产过程中使用的材料和原液储存所用的材料无浸出物风险。”4.  小结工艺相关杂质的控制是生物制品质量与安全的核心环节，需以风险分级为基础，整合上游预防、下游清除与精准检测，通过技术创新与监管协同，满足法规要求的前提下充分保证患者的安全性。下一篇将详细讨论四种特定的工艺相关杂质（HCDNA、HCP、内毒素和可浸出物）及其法规要求。