Insulin human(Eli Lilly & Co.)

摘要：大肠杆菌凭借重组 DNA 技术成为生物制药领域的核心工具，从首个重组胰岛素的生产至今，已在激素、酶、疫苗、抗体等多个领域展现重要价值。尽管存在翻译后修饰不足和毒性等局限，但其低成本、高产量和成熟的生产体系使其仍是首选宿主。本文系统梳理了大肠杆菌生产的各类生物药物，包括已获批的激素、酶、肽类、疫苗及融合蛋白等，并探讨其临床应用、优势与未来发展方向，展现其在生物制药产业中的关键地位与潜力。一、引言大肠杆菌在重组 DNA 技术的推动下，成为生物制药的重要宿主。自 1980 年代首个重组胰岛素获批以来，其凭借快速生长、低成本和高产量等优势，广泛应用于多种生物药物生产。尽管面临复杂蛋白折叠困难和缺乏糖基化修饰等挑战，但其在激素、酶、疫苗等领域的成果显著，已成为全球生物制药产业的基石之一（参考图 1: General method for producing recombinant biopharmaceuticals in Escherichia coli，了解大肠杆菌生产重组生物药物的通用流程）。二、大肠杆菌生产的生物药物类型2.1激素类药物大肠杆菌是生产重组激素的主力，涵盖胰岛素、生长激素、干扰素等。例如，首个重组人胰岛素Humulin由礼来公司开发，1982 年获 FDA 批准，彻底改变糖尿病治疗模式。随后，长效胰岛素类似物Lantus、生长激素Sogroya等相继问世，显著提升患者生活质量（表1: Approved recombinant hormones produced in Escherichia coli and their mechanism of action列出了主要激素药物及其作用机制）。干扰素 α-2b（Intron A）则用于肝炎、癌症治疗，其改良型Besremi对真性红细胞增多症疗效显著。2.2酶类药物酶类药物在代谢疾病和癌症治疗中至关重要。Krystexxa（聚乙二醇化尿酸酶）用于慢性痛风，通过降解尿酸缓解症状；Voraxaze（葡糖羧肽酶）可快速清除化疗药物甲氨蝶呤，降低肾损伤风险；Asparlas（聚乙二醇天冬酰胺酶）则用于白血病治疗，延长患者生存期（表2: Approved recombinant enzymes produced in Escherichia coli and their mechanism of action详细展示了相关酶类药物）。2.3肽类药物肽类药物在心血管和代谢疾病中应用广泛。Natrecor（奈西立肽）用于急性心力衰竭，通过扩张血管改善症状；Voxzogo（vosoritide）针对软骨发育不全，通过调节信号通路促进骨骼生长；Oxervate（神经生长因子）则用于治疗神经萎缩性角膜炎，修复角膜损伤（表3: Approved recombinant peptides produced in Escherichia coli and their mechanism of action列举了代表性肽类药物）。2.4疫苗类药物大肠杆菌在疫苗生产中表现突出，尤其在病毒样颗粒（VLP）疫苗领域。Hecolin是首个基于大肠杆菌的戊肝疫苗，在中国获批上市；Cecolin（HPV 疫苗）有效预防生殖器病变；兽用疫苗如Strangvac（马链球菌疫苗）和Leucogen（猫白血病疫苗）也展现重要价值（表4: Approved recombinant vaccines produced in Escherichia coli总结了人用及兽用疫苗）。2.5融合蛋白与抗体片段融合蛋白和抗体片段是癌症与免疫疾病的关键疗法。Lucentis（雷珠单抗）靶向 VEGF，用于湿性黄斑变性；Kimmtrak（ Tebentafusp）通过双特异性结合 T 细胞和肿瘤抗原，治疗转移性葡萄膜黑色素瘤；Cimzia（ certolizumab pegol）作为 TNF-α 抑制剂，有效缓解类风湿关节炎（表5: Approved recombinant antibodies and fusion proteins produced in Escherichia coli列出了主要抗体及融合蛋白药物）。2.6其他生物药物包括粒细胞集落刺激因子Neupogen（治疗中性粒细胞减少症）、瘦素类似物Myalept（治疗脂肪营养不良）等，均通过大肠杆菌生产，满足罕见病和免疫疾病治疗需求。三、大肠杆菌的优势与挑战优势：遗传背景清晰：便于基因操作和表达系统优化，如利用已知的克隆技术和表达载体高效生产目标蛋白。生产成本低：培养基简单（仅需基础营养物质），适合大规模发酵，例如 mRNA 新冠疫苗的低成本生产模式可复制至其他生物药。生产周期短：生长繁殖速度快，可在短时间内实现蛋白大量表达，如重组胰岛素从基因设计到生产仅需数周。挑战：翻译后修饰局限：无法进行复杂糖基化、二硫键形成等修饰，影响部分蛋白（如全长单克隆抗体）的功能，需依赖哺乳动物细胞补位（参考表5中部分抗体需哺乳动物细胞生产的限制）。内毒素风险：细胞壁成分可能引发免疫反应，需额外纯化工艺去除内毒素，增加生产成本（如疫苗生产中的严格纯化流程）。蛋白折叠问题：复杂蛋白易形成包涵体，需优化表达条件或采用共表达伴侣蛋白辅助折叠（如图1中纯化步骤的重要性）。四、未来发展方向技术优化：通过基因工程改造大肠杆菌，引入糖基化通路或共表达折叠辅助因子，提升复杂蛋白生产能力。例如，利用合成生物学技术构建 “设计型” 大肠杆菌，实现更接近天然蛋白的修饰。联合疗法拓展：大肠杆菌生产的疫苗与免疫检查点抑制剂（ICIs）联用，如mRNA 肿瘤疫苗 + PD-1 单抗的协同效应，已在黑色素瘤等适应症中展现潜力（参考前文KEYNOTE-942 研究数据）。兽用与罕见病药物：针对动物传染病（如马链球菌疫苗Strangvac）和罕见病（如脂肪营养不良药物Myalept），开发高性价比的大肠杆菌生产药物，填补市场空白。可持续生产：探索无血清培养基和连续生产工艺，降低环境负担，符合绿色制药趋势。五、结论大肠杆菌凭借低成本、高可及性和成熟的工业化体系，在生物制药领域占据不可替代的地位。尽管面临修饰能力有限等挑战，但其在激素、酶、疫苗等领域的成功案例（如表1-5中多款 FDA/EMA 批准药物）及技术优化潜力，使其持续成为创新药物开发的核心平台。未来，随着合成生物学和联合疗法的突破，大肠杆菌将进一步推动生物制药向高效、经济、普惠方向发展。识别微信二维码，添加抗体圈小编，符合条件者即可加入抗体圈微信群！请注明：姓名+研究方向！本公众号所有转载文章系出于传递更多信息之目的，且明确注明来源和作者，不希望被转载的媒体或个人可与我们联系(cbplib@163.com)，我们将立即进行删除处理。所有文章仅代表作者观点，不代表本站立场。

Taylor Tieden for BioSpace The agency’s inertia and bureaucratic roadblocks are throttling hope for millions of patients. A new center of excellence would provide a solution. When I joined the FDA to be its “biotechnology czar” more than 40 years ago, genetic engineering was still a new concept. To fully grasp the nuances of the latest technological advances, we brought together experts in different scientific disciplines from across the organization. The first “biopharmaceutical” was human insulin synthesized in genetically engineered E. coli bacteria. Despite the novelty, we decided that because the FDA had prodigious experience with insulins sourced from pig and cow pancreases and also with drugs derived from various microorganisms, recombinant DNA techniques would be viewed as an extension or refinement of long-used and familiar methods for making drugs. Thus, no sui generis policies were required. That proved to be a historic, precedent-setting–and correct—decision. The human insulin, Humulin, was approved in a record-setting five months at a time when the average approval time for a New Drug Application was 31.5 months. Biopharmaceuticals now dominate the list of highest-revenue drugs and are critical to the health of tens of millions of patients. However, when it comes to another high-impact category of drugs, those for rare diseases, the FDA has a long way to go. When it comes to approving treatments for rare diseases, the Agency’s inefficient structure creates a confusing patchwork of policies that can lead to an inconsistent review process. There are over 10,000 rare diseases, some affecting only hundreds or even just dozens of Americans. In total, more than 30 million Americans live with a rare disease, but this large, vulnerable cohort has very little visibility outside of patients’ communities and support networks. There is currently no FDA-approved treatment for an estimated 95% of rare diseases, so millions of Americans (many of whom are children) face premature mortality, disability, severe pain, repeated hospitalizations and a drastically diminished quality of life. With such a large segment of our population living in a world without cures or treatments, you would think the FDA and its parent, the Department of Health and Human Services (HHS), would be doing everything in their power to usher in a new wave of innovation. But, perhaps unsurprisingly in government bureaucracies, they lack focus, coordination and an overarching strategy for addressing rare diseases. To become more effective in this area and to give hope to patients living on borrowed time, the FDA should establish a cross-cutting entity with a single leader to focus resources on the goal of getting treatments to rare-disease patients who need them without unnecessary delays. The FDA is Throttling Innovation In 1983 the Orphan Drug Act, which incentivized researchers to invest in new drugs and devices for rare diseases, was enacted. Before that landmark law was passed, just 38 rare disease drugs had been approved by the FDA; today, that number is well over 1,000. This is good news for patients, but much more must be done. The FDA continues to add and move boxes around on its organizational chart, but rare diseases continue to get short shrift, and innovation has suffered. Rare diseases are lumped into the Office of Rare Diseases, Pediatrics, Urologic and Reproductive Medicine, which dilutes both the expertise and focus on rare diseases. A further complication is that many therapies for rare diseases are classified as “biologics,” which are regulated not by the Center for Drug Evaluation and Research but by the Center for Biologics Evaluation and Research. Different groups of bureaucrats may have different biases and approaches, which causes the goalposts for approval to be constantly shifted, leading to additional, costly, difficult-to-perform new trials. This situation arises because expertise in the highly specialized fields required to understand the spectrum of rare diseases and the mechanisms for treating them is scattered across the Agency. The conflicting guidance and advice given by different components of the FDA is not a prescription for efficiency and expert oversight, let alone advocacy for rare disease drugs. And the involvement of regulators who advocate for the expeditious testing and approval of the drugs is important: There is a saying in government that “Personnel = Policy.” Unfortunately, many pharmaceutical companies decide it’s simply not worth the hassle of the additional regulatory burden, or that they can’t afford the cost of providing drugs on a “compassionate use” basis and gathering new data. Consider these recent examples: In April, after years of deliberations, the FDA finally agreed to consider a drug to help an estimated patient population of 130–150 Americans. The review had been delayed after the agency gave the drugmaker conflicting requirements for data it needed to submit, and prior to the FDA’s acceptance of its application, the company had planned to abandon the drug. The same month, a company founded a quarter-century ago went bankrupt after the FDA kept moving the goalposts on its treatment for a disease that affects one in four million newborns and has been shown to extend children’s lives. (Inexplicably, the agency also failed to grant the company an approval or even an Emergency Use Authorization for its drug to treat COVID .) A Call to Action To prevent unnecessary roadblocks to vital medicines, the FDA needs an organization that understands and will focus on advocacy for rare-disease drugs. Last year, the FDA made a step in the right direction when it launched the Support for clinical Trials Advancing Rare disease Therapeutics (START) Pilot Program to accelerate “the pace of development of certain CBER- and CDER-regulated products (novel drug and biological products) that are intended to treat a rare disease.” To achieve this, the agency said: Selected [drug developers] will be able to interact with FDA through rapid, ad-hoc communication mechanisms to provide a mechanism for addressing program-specific development issues, including but not limited to clinical study design, choice of control group, fine-tuning the choice of patient population, leveraging nonclinical information, or product characterization. The FDA’s recent establishment of a Rare Disease Innovation Hub is another noteworthy advance in addressing the needs of patients with rare diseases, but more is needed. Building on these efforts, the FDA should now create a new organization resembling the agency’s Center of Excellence for Oncology to focus on getting more treatments to patients who have precious little time or options. Developing a Center of Excellence (or “Intercenter Institute”) for Rare Diseases under a single, dedicated director would further streamline rare disease development. This idea, which has the support of bipartisan members of Congress , would aggregate specialists and researchers on different types of rare diseases and organ systems to expedite the testing, evaluation and approval of drugs for rare diseases. Ultimately, it would enhance cross-Agency collaboration and expedite drug delivery to patients who currently lack options for treatment. Many of these treatments will be cutting-edge gene and cell therapies and combination therapies that require expertise across multiple disciplines to fully understand how they work. Institutional change won’t be easy, but it is necessary. A Center for Excellence for Rare Diseases would be an excellent start.