This is a single-center, prospective, randomized, controlled (crossover) clinical study designed to investigate the impact of lowering insulin levels on hepatic glucose production (HGP) vs de novo lipogenesis (DNL) in people with insulin resistance. The investigators will recruit participants with a history of overweight/obesity and evidence of insulin resistance (i.e., fasting hyperinsulinemia plus prediabetes and/or impaired fasting glucose and/or Homeostasis Model Assessment of Insulin Resistance [HOMA-IR] score >=2.73), and with evidence of metabolic dysfunction-associated steatotic liver disease (MASLD). Participants will undergo two pancreatic clamp procedures -- one in which serum insulin levels are maintained near hyperinsulinemic baseline (Maintenance Hyperinsulinemia or "MH" Protocol) and the other in which serum insulin levels are lowered by 50% (Reduction toward Euinsulinemia or "RE" Protocol). In both clamps the investigators will use stable-isotope tracers to monitor hepatic glucose and triglyceride metabolism. The primary outcome will be the impact of steady-state clamp insulinemia on HGP vs DNL.

开始日期2025-06-01

申办/合作机构

Columbia University

[+2]

NCT06434038

/ Not yet recruiting临床1期IIT

Measurement of Insulin Levels in the Cerebrospinal Fluid (CSF) of Healthy Adults After a Single Intranasal Dose - Middle Age

The purpose of this study is to find out whether insulin, a drug approved by the FDA for the treatment of diabetes mellitus, reaches the brain and spinal cord when delivered as a nasal spray (intranasally). Intranasal insulin has been shown to improve memory and mood in patients with neurological diseases such as mild cognitive impairment and dementia, but more evidence is needed to support the ability to effectively target the brain through intranasal routes.
18 healthy middle-aged adults will be randomly assigned to receive a single intranasal dose of 40 units insulin ("low dose" group), 80 units insulin ("high dose" group), or saline (placebo, or control group). Participants will undergo an image-guided lumbar puncture (spinal tap) performed by a study clinician. Samples of cerebrospinal fluid (a fluid surrounding the brain and spinal cord) and blood will be collected at 5 timepoints during the lumbar puncture: once prior to the administration of intranasal insulin, and again at 10, 20, 30, and 40 minutes after the dose is given. Samples will be tested to determine the level of insulin detected in the cerebrospinal fluid and blood at each time point. Results of this study will provide essential information about the ability of insulin to reach the brain after intranasal administration.

开始日期2025-03-31

申办/合作机构

Health Partners Institute For Education & Research

[+2]

NCT04809311

/ CompletedN/A

Fully Decentralised Clinical Study Exploring Remote Collection of Glycaemic and Behaviometric Data Among Participants With Type 2 Diabetes Mellitus on Different Treatment Regimens

Earlier protocol ID was NN1250-4630, protocol ID is changed to NN1535-7774. The purpose of this study is to collect blood glucose values and activity data in patients with type 2 diabetes for approximately 12 weeks using electronic devices in a full virtual clinical setting. Virtual clinical setting means that all data are collected by use of participants' personal smartphone and study-related apps. In other words, everything will be handled without any visits to a hospital or doctor.
For collection of participants' blood glucose values participants will receive two different blood glucose monitors. One where participants cannot see the blood glucose values and one where participants can see the blood glucose values. Participants must wear them consecutively for 2 and 10 weeks, respectively. Further, participants will be asked to self-apply the monitors on their upper arm.
For collection of participants' activity data participants will receive an activity tracker, which participants will wear on the wrist throughout the study.
The study does not include any study medication and participants will continue the current antidiabetic treatment as prescribed to participants by their own physician. If any questions about the treatment and/or health condition while participating in the trial, participants should consult your own physician If participants are in doubt about what the blood glucose values mean or whether participants should react to the blood glucose values, participants need to contact the research staff or their general practitioner.

开始日期2024-04-30

申办/合作机构

Novo Nordisk A/S

100 项与人胰岛素（诺和诺德）相关的临床结果

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347

项与人胰岛素（诺和诺德）相关的文献（医药）

2025-04-01·JOURNAL OF CONTROLLED RELEASE

BioJect: An in vitro platform to explore release dynamics of peptides in subcutaneous drug delivery

Article

作者: Qin, Qiuhua ; Wacker, Matthias G ; Kahouadji, Lyes ; Benetti, Ayça Altay ; Li, David

Predicting the release performance of subcutaneous (SC) drug formulations is challenging due to the complex interplay between physicochemical properties and the physiological microenvironment, which includes the extracellular matrix (ECM), fluid composition, and fluid availability, factors that collectively influence bioavailability and absorption rates. The ECM often acts as a bandpass filter modulated by local ion and protein content. In this study, we introduce the BioJect cell, a modern release test method based on the compendial flow-through cell, integrating a perfusion system with customizable biomatrix components. We systematically investigated the release mechanisms of four insulin formulations: regular human insulin, insulin aspart, insulin glulisine, and Neutral Protamine Hagedorn (NPH) insulin. A modified simulated subcutaneous interstitial fluid (mSSIF) comprising multiple components of the SC physiological environment was employed. It incorporates important ions and proteins (138.5 mM sodium, 10 mM potassium, 1.8 mM calcium, 0.8 mM magnesium, 111.3 mM chloride, 28 mM bicarbonate, 0.5 mM sulfate, 5 mM acetate, 4.2 mM phosphate, 30 g/L total protein added as bovine serum albumin). Our release test method discriminated the tested formulations under varying biorelevant conditions, demonstrating its biopredictive capabilities. Notably, we discovered a previously undocumented albumin binding affecting the release rate of insulin glulisine, likely occurring in the low-shear environment of SC tissue only. Additionally, the inclusion of biorelevant components like hyaluronic acid and collagen into the biomatrix of the BioJect cell provided profound insights into potential absorption and release mechanisms, supported by two in vitro-in vivo relationships (level C and level A). The BioJect cell represents a significant advancement in simulating the SC environment for drug release testing. Our findings highlight the importance of considering protein binding and ECM components in predicting drug absorption, offering a promising tool for the development and optimization of SC formulations.

2025-03-01·CLINICAL AND EXPERIMENTAL OPHTHALMOLOGY

Drug‐associated glaucoma: A real‐world study based on the Food and Drug Administration adverse event reporting system database

Article

作者: Guan, Wen‐Ying ; Liu, Zuguo ; Hu, Jiaoyue ; Wang, Yu‐Qian ; Huang, Caihong ; Chen, Xiao‐Dong ; Wu, Shi‐Nan ; Wang, Shao‐Pan ; Yan, Dan

Abstract:

Background:

This study aims to assess the risk of drug‐associated glaucoma and track its epidemiological characteristics using real‐world data.

Methods:

Adverse event reports from the Food and Drug Administration Adverse Event Reporting System (FAERS) from January 2004 to December 2023 were analysed. Disproportionality analysis and the Bayesian Confidence Propagation Neural Network algorithm were used. The study classified drugs associated with glaucoma, assessed risk levels, and compared drug‐induced times across different categories.

Results:

Eight hundred and five drugs were linked to glaucoma in the FAERS database. Disproportionality analysis identified 46 drugs with significant risk, mainly adrenergic medications (clobetasol propionate, fluocinolone acetonide), antihypertensives (hydrochlorothiazide), insulin (insulin human), anticholinergics (umeclidinium, darifenacin), VEGF inhibitors (brolucizumab, faricimab), and psychotropics (topiramate, ziprasidone). The top three high‐risk drugs were clobetasol propionate, umeclidinium, and fluocinolone acetonide. The shortest drug‐induced times were observed with indacaterol, salmeterol, and umeclidinium. Anticholinergic medications had the shortest drug‐induced time among all categories. Females (62.5%) and the elderly (average age 63.5 ± 16.8 years) were predominantly affected. Reports of drug‐associated glaucoma increased over the years.

Conclusion:

Preventing drug‐associated glaucoma is more effective than treatment. Identifying the risk and drug‐induced times of systemic and ophthalmic drugs can reduce occurrence risk. Clinical practitioners should be vigilant and inform patients of these risks.

2024-12-02·ChemMedChem

Higher Order Structure Differences Among Insulin Crystalline Drugs Revealed by 2D heteronuclear NMR

Article

作者: Rienstra, Chad M. ; Chen, Kang ; Wang, Songlin

Abstract:

During therapeutic protein development, two‐dimensional (2D) heteronuclear NMR spectra can be a powerful analytical method for measuring protein higher order structure (HOS) in solution since the spectra exhibit much higher resolution than homonuclear 1H spectra. However, 2D NMR capabilities for characterizing protein HOS in crystalline states remain to be assessed, given the low 13C natural abundance and intrinsically broader lines in solid‐state NMR (SSNMR). Herein, high‐resolution heteronuclear correlation (HETCOR) SSNMR was utilized to directly measure intact crystal drug products of insulin human, insulin analogs of insulin lispro and insulin aspart. The fingerprint regions in 2D 1H−13C HETCOR spectra were identified, which distinguished the insulin crystals in their primary structure, HOS heterogeneity and dynamics, as well as the manufacturing processes. The HOS heterogeneity in insulin analogs is consistent with their therapeutic effect of rapid action; while insulin human crystals showed more structural homogeneity, consistent with their slower pharmacokinetics (PK) peak time than insulin analogs. Therefore, heteronuclear NMR could be broadly applicable to study protein drug dosage forms from liquid to solid, yielding improved molecular level structure data for assessing drug HOS in biosimilar drug development.

项与人胰岛素（诺和诺德）相关的新闻（医药）

2025-03-01

·医药投资部落

国产胰岛素龙头的十年一梦

2015年3月，通化东宝股价最高摸到8.84元/股。 2025年3月，通化东宝最新收盘价格为7.45元/股。在胰岛素这个号称长坡厚雪的赛道上，作为国产龙头企业的通化东宝，十年的时间恍如黄粱一梦。 2024年1月，通化东宝披露2024年业绩预告显示，公司预计2024年年度实现归属净利润约为4052.77万元，同比减少约96.53%。公开数据显示，这将是通化东宝自2007年以来最差的业绩。胰岛素集采是造成通化东宝净利大幅下滑的重要原因之一。通化东宝表示，2024年新一轮胰岛素集采在全国各省市开始陆续落地实施，公司各胰岛素产品中标价格均有不同程度下降，导致销售收入下滑。同时，由于新一轮胰岛素集采落地带来公司产品价格下调的普遍预期，公司商业客户对库存进行控制与调整，对公司发货造成一定影响，因此销售收入同比下滑。与集采如影随形的，是同行之间的激烈竞争。 2024年2月26日，通化东宝发布2024年业绩预告更正公告：经财务部门再次测算，预计2024年实现归属于上市公司股东的净利润约为1164.21万元，同比减少约99%。业绩预告更正的主要原因，是通化东宝与同行甘李药业之间的官司。 2021年1月，甘李药业起诉通化东宝生产销售的“长舒霖”甘精胰岛素，侵害其“长秀霖”商标权，并构成不正当竞争。根据最终判决，通化东宝应赔偿甘李药业经济损失6000万元。导致通化东宝2024年业绩大亏的，还有终止的研发管线。 2024年7月11日，通化东宝发布公告称，经审慎考量，公司决定终止可溶性甘精赖脯双胰岛素注射液研发项目（以下简称“THDB0207注射液”）临床研究开发工作。可溶性甘精赖脯双胰岛素注射液，是一种创新的胰岛素制剂，它结合了长效的甘精胰岛素和速效的赖脯胰岛素，与预混胰岛素相比，甘精赖脯双胰岛素能够强化空腹血糖控制，并减少“肩效应”引发的低血糖风险。在6年前的2018年4月26日，通化东宝公司与法国SA ADOCIA公司签订许可协议，通化东宝有偿获得BioChaperone® Combo（胰岛素基础餐时组合）在所有大中华地区，以及马来西亚、新加坡等国家和地区的独家开发、生产并商业化权利。 2022年5月，通化东宝先后启动了CT046、CT047、CT048三项I期临床试验，研究主要目的在于评估THDB0207注射液的临床药理学特征。 2023年10月，通化东宝收到上述三项关键I期临床试验的总结报告，研究结果显示达到主要终点目标。 2024年二季度，在I期临床试验的基础上，综合多位中国权威临床专家的意见，得出需要补充开展II期临床试验的结论。主要原因是，基于THDB0207注射液I期临床试验的数据，在中国最新临床实践下，有必要继续探索该产品的临床价值，其中对夜间低血糖特征的充分考察尤为重要。但是这个时候，通化东宝的这款产品，已经完全不占市场先机。目前，在国内已上市的与THDB0207注射液类似的双胰岛素注射液，有诺和诺德公司于2019年5月获批上市的德谷门冬双胰岛素。此外，还有大批在研管线：吉林惠升的德谷门冬双胰岛素生物类似药于2023年已经报产，联邦制药、东阳光、重庆宸安、中美华东这4家公司的德谷门冬双胰岛素生物类似药已于2022年至2024年期间进入III期临床试验阶段，其研发进度都大幅领先于通化东宝的THDB0207注射液。在这种不利局面下，通化东宝做出了终止研发的决策。通化东宝项目研发资本化金额6455.82万元全额计提资产减值准备，同时对2018年已经支付给SA ADOCIA公司与该项目相关的预付商业化权利款金额2.54亿元确认为损失。上述两项会计处理，将减少公司2024年年度净利润2.70亿元。应该说，2024年的通化东宝，运气实在有点差。但是好转的迹象也正在显露。一方面，在最新一轮胰岛素集采中，公司胰岛素产品尽管价格有所下降，但全部胰岛素产品均成功以A/A1类中标，除基础签约量外，还获得了约1,000万支分配量，并在医院准入方面实现巨大的提升和突破。另外一方面，公司的出海战略正在取得成绩。 2024全年，通化东宝的海外业务大幅增长，预计实现收入超亿元，同比接近翻倍。通化东宝的人胰岛素注射液上市许可申请已获得EMA正式受理；同时公司与健友股份已就门冬、甘精、赖脯3款胰岛素达成战略合作，共同进军美国胰岛素市场，目前相关临床试验正在稳步推进中。此外，在创新药领域，公司进展也不小。 2月25日，通化东宝宣布完成了注射用THDBH120的一项I期临床试验，并获得临床试验总结报告，试验达到主要终点目标。注射用THDBH120是通化东宝重点在研的胰高血糖素样肽-1 (GLP-1) 受体和葡萄糖促胰岛素分泌多肽 (GIP) 受体双靶点激动剂，属于一类创新药，有望成为更长效的治疗糖尿病和肥胖的重磅药物。医药研究报告分享营收集与分享全市场主流医药研究报告每天仅需0.8元畅享超过2000篇存量精选医药研究报告每年新增额外500篇

财报IPO专利侵权临床1期

2025-02-06

·echemi

Novo Nordisk Reports 2024 Earnings with $42.13 Billion Revenue and Impressive Profit Margins

On February 5, Novo Nordisk officially released its annual report for 2024, revealing a remarkable revenue of 29.04 billion Danish kroner (approximately $42.13 billion), representing a 25% increase year-on-year. The operating profit also surged to 12.83 billion Danish kroner (around $18.62 billion), reflecting a 25% growth, while net profit reached 10.10 billion Danish kroner (about $14.65 billion), up 21%. Notably, Novo Nordisk's profit margin (operating profit/sales) stands at an impressive 44.19%, far surpassing many American pharmaceutical giants. Novo Nordisk operates primarily in two business segments: Diabetes and Obesity Care and Rare Diseases. The majority of its revenue comes from the Diabetes and Obesity Care segment, which generated 27.18 billion Danish kroner in sales for 2024, while the Rare Diseases segment contributed only 1.86 billion Danish kroner. The GLP-1 class of drugs has emerged as Novo Nordisk's biggest revenue driver. Rybelsus (oral semaglutide) achieved sales of 23.30 billion Danish kroner (approximately $3.38 billion), while Ozempic (injectable semaglutide) brought in 120.34 billion Danish kroner (around $17.46 billion). The weight-loss version, Wegovy, also performed well with sales of 58.21 billion Danish kroner (about $8.44 billion). The total sales for semaglutide reached $29.28 billion, showcasing its strong performance, although it has not yet dethroned Keytruda as the new king of drugs. In addition to semaglutide, Novo Nordisk's GLP-1 offerings include the previous generation liraglutide, with Victoza (liraglutide for diabetes) and Saxenda (liraglutide for weight loss) generating 5.48 billion Danish kroner (approximately $795 million) and 6.94 billion Danish kroner (around $1.01 billion), respectively. Insulin remains Novo Nordisk's second-largest revenue generator, with total sales of 55.37 billion Danish kroner (approximately $8.03 billion) in 2024, driven by the success of degludec insulin, which saw a 15.3% increase in sales compared to the previous year. The three long-acting insulins—Tresiba (degludec), Xultophy (degludec/liraglutide), and Levemir (detemir)—achieved sales of 9.91 billion, 4.67 billion, and 4.50 billion Danish kroner, respectively. The two premixed insulins, Ryzodeg (degludec/aspart) and NovoMix (premixed aspart), generated revenues of 4.93 billion and 5.86 billion Danish kroner, respectively, with the former showing growth while the latter declined. Rapid-acting insulins, including Fiasp (ultra-rapid aspart) and NovoRapid (aspart), collectively sold 18.52 billion Danish kroner, an increase of approximately 2.6 billion compared to the previous year. Additionally, Novo Nordisk continues to sell second-generation human insulin, which generated 6.97 billion Danish kroner, although this segment saw a year-on-year decline. In the Rare Diseases segment, Novo Nordisk's product pipeline primarily includes clotting factors and growth hormones, with total sales of only 1.86 billion Danish kroner, representing a small portion of overall revenue. Unlike other major international pharmaceutical companies, Novo Nordisk has not pursued large-scale acquisitions and maintains a research and development investment intensity of around 15%. However, the company's sustained high revenue growth demonstrates its efficiency in product development. This efficiency can be attributed to four key factors: a strong focus on diabetes, extensive knowledge accumulation over a century, a commitment to user experience, maximizing market value from existing products, and the development of platform technologies. With advanced fatty acid acylation technology, peptide oral delivery technology, and yeast fermentation technology, Novo Nordisk is well-positioned to maintain its competitive edge in the market.

并购财报上市批准生物类似药

2025-01-20

·药时空

一文拿捏 | 从大肠杆菌包涵体中获取重组人胰岛素的下游纯化工艺

导读近年来全球糖尿病患者数量持续上升，对胰岛素的需求不断增加，而经济实惠的胰岛素供应不足，因此开发高效经济的生产工艺至关重要。重组人胰岛素生产主要有大肠杆菌包涵体和酵母表达系统两种途径，大肠杆菌因生长快、成本低等优势成为主要宿主，但包涵体路线需复杂的下游处理。新加坡研究人员曾在Bioresources and Biprocessing上发表了题目为《Downstream processing of recombinant human insulin and its analogues production from E. coli inclusion bodies》的综述。重点阐述了从大肠杆菌包涵体生产重组胰岛素下游加工的各个环节，并对每个环节的方法、条件和影响因素进行了详细讨论。从大肠杆菌包涵体中纯化重组人胰岛素/类似物的下游工艺很复杂，涉及多个步骤，包括包涵体回收、洗涤、溶解和氧化亚硫酸化、氰化溴裂解、缓冲液交换、色谱纯化、pH沉淀和锌结晶、胰岛素原折叠、酶切和制剂等，且每个步骤中采用的方法对下一步的产品纯度和产量产生重大影响。图1 重组人胰岛素及其类似物下游加工的一般工作流程一、包涵体回收与洗涤由于异源蛋白在大肠杆菌中以不溶性细胞质包涵体形式存在且不分泌到培养基，所以需要特定方法进行回收。破坏细胞包膜的机械方法包括超声处理、在胶体磨中研磨细胞悬浮液或通过Manton-Gaulin高压等方式破碎细胞膜，该方法能有效地破坏细胞，但可能会更严重地损害包涵体中的蛋白质质量[1]。在基于溶菌酶的方法中，添加溶菌酶以消化细胞包膜。溶菌酶可裂解肽聚糖的骨架，使细胞结构被破坏，随后通过膜微滤或离心技术从全细胞裂解物中分离出包涵体。常用的洗涤缓冲液添加剂有尿素、Triton X-100、EDTA、甘油、溶菌酶、NaCl、Tween 20等，其中尿素和Triton X-100最为常见。大肠杆菌细胞壁由磷脂、蛋白质、肽聚糖和脂多糖组成，使用去污剂、低浓度尿素、溶菌酶和EDTA进行选择性萃取有助于去除细菌细胞壁成分。Triton X-100能去除膜磷脂和碎片，促进碎片和可溶性蛋白从包涵体上解离。洗涤优化要确保洗涤缓冲液不会溶解所需的重组蛋白，此外离心次数、速度和洗涤温度等参数也需要优化，以实现时间和成本的有效控制，提高生产效率。图2 蛋白质-添加剂的相互作用模式二、包涵体溶解及氧化亚硫酸化包涵体通常使用高浓度变性剂，如胍盐酸盐和尿素能完全破坏蛋白质结构，对于含多个半胱氨酸残基的蛋白质，可添加二硫苏糖醇（Dithiothreitol ，DTT）或β-巯基乙醇（Mercaptoethanol，BME）防止错误二硫键形成。在正确的构象中，硫解步骤对于保持其展开形式至关重要，防止在溶解过程中和蛋白质复性之前的初始纯化步骤中形成可能不正确的二硫键，此外氧化硫化分解有助于提高后续复性产量。pH值和温度是可能影响反应溶解和硫解速率的因素，为了获得最大产量，监测硫解反应的温度和持续时间以确保反应完成非常重要。三、溴化氰裂解及缓冲液交换在大肠杆菌蛋白质翻译过程中，重组蛋白以融合蛋白形式表达，N端起始氨基酸为N-甲酰甲硫氨酸（N-Formylmethionine，fMet）。fMet会被人体免疫系统识别为异物，可能引发不必要的免疫原性反应，因此在纯化前需将其去除，溴化氰裂解就是用于切断胰岛素原的甲硫氨酸连接子，从而去除fMet这一信号序列。该方法的缺点是切割特异性低，溴化氰蒸发时间长，其高毒性和挥发性以及释放产物可能发生化学修饰。另一种方法是使用蛋白质蛋白酶来切割融合蛋白，但必须确保蛋白酶在正确的位点切割以去除N末端融合肽。缓冲液交换能去除影响蛋白功能和稳定性的试剂，使蛋白适应后续操作，主要通过三种方法实现。透析：利用渗透压使溶质透过半透膜，是传统的脱盐和缓冲液交换方法。适合易沉淀的敏感蛋白，但耗时久且需较大的膜表面积。超滤：借助离心力或其他外部压力使小分子溶质透过多孔膜，而大分子溶质被截留，从而实现脱盐和缓冲液交换，同时能浓缩蛋白样品。在工业生产中，超滤常用于包涵体溶解和亚硫酸化反应后，去除高浓度尿素和亚硫酸化试剂；在蛋白复性后，也能将复性样品浓缩并交换缓冲液，为后续的酶促转化提供合适环境。尺寸排阻色谱：基于分子大小差异进行分离，小分子如盐类可与较大的肽段分离，是透析的快速替代方法，稀释因子低。四、色谱纯化与沉淀 4.1 色谱纯化亲和色谱（Afnity Chromatography，AC）：基于配体与目标蛋白的特异性结合，需在胰岛素原分子上融合合适的标签。融合标签不仅有助于纯化，还能提升胰岛素原稳定性和表达水平，但基因融合在下游处理时，需考虑标签的去除及对后续分析的影响，且AC树脂成本较高。离子交换色谱（Ion‑Exchange Chromatography，IEX）：根据分子电荷差异分离，可选用阴离子交换色谱或阳离子交换色谱，胰岛素及其前体具有可离子化基团，IEX可用于纯化折叠或磺化的胰岛素原。反相色谱（Reversed‑phase Chromatography，RP）：依据分子与固定相的疏水相互作用分离，通常在胰岛素原酶促转化为胰岛素后使用。RP能有效去除胰岛素原未充分裂解或错切产生的杂质，C8柱因胰岛素的分子量和疏水性成为常用选择，常用乙腈作为洗脱剂[2]。疏水相互作用色谱（Hydrophobic Interaction Chromatography，HIC）：与RP类似基于疏水性分离，但HIC吸附剂疏水性较弱，洗脱无需有机溶剂，对敏感蛋白更温和。尺寸排阻色谱（Size Exclusion Chromatography，SEC）：根据分子大小分离，是较温和的技术，常用于胰岛素纯化的抛光步骤，也可在蛋白质折叠液相色谱中与复性过程结合。混合模式色谱（Mixed‑Mode Chromatography，MMC）：利用多种相互作用实现分离，在胰岛素纯化中逐渐受到关注，其混合模式离子交换剂与传统离子交换剂相比具有不同选择性，可在高盐条件下结合蛋白质，减少脱盐或稀释步骤。表1 胰岛素原（PI）纯化中的亲和色谱法 4.2 沉淀方法 pH沉淀：利用蛋白质在等电点时溶解度最低的特性，通过调节溶液pH使胰岛素/胰岛素类似物沉淀，从而与杂质分离，还能浓缩蛋白。例如可将溶解的融合蛋白pH调至5.0使其沉淀，离心后再溶解于碱性缓冲液提高蛋白纯度；在胰岛素原复性后，也可调节pH沉淀杂质，收集含正确折叠胰岛素原的上清。锌结晶：胰岛素在锌离子存在下可形成六聚体等聚集体，利用这一特性在纯化过程中加入锌离子使胰岛素结晶，可用于胰岛素的浓缩和临时保存，减少其在储存过程中的物理和化学降解[3]。五、复性胰岛素由A、B两条多肽链通过二硫键连接而成，胰岛素原在体外复性时，需去除高浓度变性剂，促使二硫键正确形成和蛋白质折叠为天然结构。在复性缓冲液中稀释溶解的蛋白质是胰岛素生产中最常用的复性方法，其他回收胰岛素原的方法包括蛋白质折叠液相色谱法（Protein-Folding Liquid Chromatography，PFLC）和透析。PFLC具有连续复性、生产率高和缓冲液消耗低等优势，但设备成本较高。透析法耗时较长，且在大规模生产中应用存在一定局限性。为了提高胰岛素原复性得率，可以在复性缓冲液中添加离液剂，同时缓冲液成分、氧化还原试剂、蛋白质浓度、温度、时间及融合标签等都会影响复性效率。表2 通过稀释法进行胰岛素原（PI）重折叠的优化条件六、酶促切割及制剂在生产“胰岛素原”途径中，需通过酶促切割去除重组人胰岛素/类似物前体分子中的C肽，以形成具有活性的胰岛素异二聚体。常使用胰蛋白酶和羧肽酶B（Carboxypeptidase B，CPB）进行切割，胰蛋白酶是丝氨酸内肽酶，能切割内部精氨酸和赖氨酸残基的羧基末端；CPB作为外肽酶可去除胰蛋白酶作用后C末端的精氨酸、赖氨酸和组氨酸等碱性氨基酸。可通过RP-HPLC监测酶促反应确定反应终点。反应终止方法多样，如将反应混合物pH调至2.5-3.5适合直接上阳离子交换色谱；添加蛋白酶抑制剂如抑肽酶，适用于后续为阴离子交换色谱/混合模式色谱的情况。在完成最终的色谱纯化后，胰岛素分子面临脱酰胺和聚集/纤维化的风险。为防止这些情况发生，需通过结晶和冻干去除残留的盐和缓冲液。胰岛素在锌离子存在下可形成晶体，通过离心、倾析或过滤分离胰岛素晶体，并洗涤去除残留锌。锌结晶后进行冷冻干燥，使胰岛素在储存时保持稳定，在使用前再将其溶解。为了抑制细菌生长、防止蛋白质聚集或改变胰岛素的体内吸收动力学，会向制剂中添加特定添加剂。间甲酚可抑制胰岛素淀粉样蛋白形成；胰岛素与锌和/或鱼精蛋白络合，能够制备中效或长效制剂，满足不同的临床需求。结语本文全面介绍了大肠杆菌包涵体生产重组人胰岛素的下游加工全过程，该过程步骤繁多，各环节紧密相连且相互影响，并提供了关键生产阶段实例，讨论了多模式纯化方案整合的实际问题，旨在为生产更高效、经济的胰岛素提供指导，以应对全球糖尿病患者对胰岛素的庞大需求。未来需优化整合各步骤，开发更高效、经济的生产工艺，以满足全球对胰岛素的需求，降低患者治疗成本。参考文献 [1] Singhvi P, Saneja A, Srichandan S, et al. Bacterial Inclusion Bodies: A Treasure Trove of Bioactive Proteins. Trends Biotechnol. 2020 May; 38(5):474-486. [2] Govender K, Naicker T, Baijnath S, et al. Sub/supercritical fluid chromatography employing water-rich modifier enables the purification of biosynthesized human insulin. J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Oct 15;1155:122126. [3] Zieliński M, Romanik-Chruścielewska A, Mikiewicz D, et al. Expression and purification of recombinant human insulin from E. coli 20 strain. Protein Expr Purif. 2019 May;157:63-69. 识别微信二维码，可添加药时空小编请注明：姓名+研究方向！

微生物疗法

100 项与人胰岛素（诺和诺德）相关的药物交易

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外链

KEGG	Wiki	ATC	Drug Bank
D03230	-	A10AF01 A10AD01 A10AC01	DB00030

研发状态

批准上市

10 条最早获批的记录,

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适应症	国家/地区	公司	日期
1型糖尿病	美国	Novo Nordisk, Inc.	1991-06-25
2型糖尿病	美国	Novo Nordisk, Inc.	1991-06-25
高血糖	美国	Novo Nordisk, Inc.	1991-06-25
糖尿病	美国	Novo Nordisk, Inc.	1983-08-30

未上市

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适应症	最高研发状态	国家/地区	公司	日期
脓毒性休克	临床3期	德国	Novo Nordisk A/S	2003-04-01
脓毒性休克	临床3期	德国	B. Braun Melsungen AG	2003-04-01

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临床结果

适应症

分期

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研究	分期	人群特征	评价人数	分组	结果	评价	发布日期


NCT04115384 (CTgov)	临床2期	额颞痴呆	3	Novolin-R insulin	築構鑰糧選網鏇鏇齋壓 = 壓壓糧艱壓衊鑰糧製網鹹憲餘鏇鹹淵積醖鹹蓋 (鹹衊憲鏇網糧蓋餘繭醖, 夢鑰鹹壓廠鑰糧醖遞憲 ~ 憲艱製獵齋鬱廠鬱鑰醖) 更多	-	2023-07-06
NCT02415556 (MemAID, CTgov)	临床2期	非胰岛素依赖型糖尿病2 \| 2型糖尿病	289	Regular Human Insulin (Type 2 Diabetes Mellitus - Insulin)	廠選鹽鑰繭膚獵艱繭蓋(齋構衊淵壓夢糧蓋蓋獵) = 壓選襯鏇選鹽餘鏇構醖選蓋願簾繭夢願襯窪繭 (網鹽憲顧範淵製窪簾壓, 22.94) 更多	-	2021-09-17
				Placebo (Type 2 Diabetes Mellitus - Placebo)	廠選鹽鑰繭膚獵艱繭蓋(齋構衊淵壓夢糧蓋蓋獵) = 範積蓋糧膚顧淵積膚衊選蓋願簾繭夢願襯窪繭 (網鹽憲顧範淵製窪簾壓, 21.04) 更多
NCT03511521 (CTgov)	临床4期	胰岛素抵抗 \| 类固醇诱导的高血糖	3	Glargine+Humulin N+Insulin Aspart+Novolin N (NPH Insulin)	憲衊蓋鏇觸築餘網糧壓(觸壓艱簾蓋構糧遞鑰膚) = 鹽餘餘製壓襯憲築鏇顧繭鑰鑰餘蓋蓋襯範顧繭 (積夢顧簾襯構艱鬱遞積, 81.0) 更多	-	2020-09-11
				Glargine+Insulin Aspart (Basal/Bolus Insulin)	憲衊蓋鏇觸築餘網糧壓(觸壓艱簾蓋構糧遞鑰膚) = 構淵夢鑰醖觸夢淵憲觸繭鑰鑰餘蓋蓋襯範顧繭 (積夢顧簾襯構艱鬱遞積, 61.8) 更多
NCT02138006 (SDIS, CTgov)	N/A	1型糖尿病 \| 脑卒中 \| 肾功能不全 ... 更多	102	Monotard (Intensive Insulin Treatment)	鑰願膚繭壓鏇遞築憲製 = 窪醖齋觸糧憲襯繭憲簾獵艱蓋窪憲襯範鹽鬱積 (憲鑰觸壓壓觸構觸襯醖, 鏇鏇齋窪繭範觸艱壓鏇 ~ 構憲鬱網願構顧壓糧糧) 更多	-	2015-12-21
				Standard insulin treatment (Standard Insulin Treatment)	鑰願膚繭壓鏇遞築憲製 = 願蓋鏇鏇願繭憲憲顧構獵艱蓋窪憲襯範鹽鬱積 (憲鑰觸壓壓觸構觸襯醖, 餘衊鏇製積築繭願憲積 ~ 窪鏇鏇範願顧鹹膚衊艱) 更多