Two-thirds of the US population, particularly African Americans (AA), is at risk for inadequate or deficient 25-hydroxy-vitamin D (25(OH)VD). Epidemiological studies demonstrate an association between better health outcomes and higher blood levels of 25(OH)VD . Randomized controlled clinical trials have shown that, while supraphysiological high doses of VD are needed to achieve adequate blood levels of 25(OH)VD, not all subjects respond to them. Recent studies have also questioned the therapeutic effects of high-dose VD supplementation. Severe VD deficiency has been associated independently with the future risk of mild cognitive impairment (MCI) and dementia. A reduction in GSH and an increase in the oxidative stress levels of serum, erythrocytes, and circulating lymphocytes has been observed in MCI and Alzheimer disease, findings similar to those in VD deficient persons. Scholarly reviews conclude that excess oxidative stress is one of the major risk factors for AD and support a potential therapeutic role for L-cysteine (LC, a GSH precursor) and vitamin D (VD) supplementation in the treatment of Alzheimer disease symptoms. This application presents the investigators' design for a randomized, double-blind, placebo-controlled clinical trial to test the hypothesis that supplementation with VD in combination with L-cysteine (LC) is more successful at optimizing the statuses of 25(OH)VD [biological signatures] and simultaneously decreasing TNF-α, IR [functional or clinical outcomes], and oxidative stress, suggesting a better therapeutic approach compared with supplementation with VD alone in AA subjects.

开始日期2020-11-06

申办/合作机构

LSU Health Sciences Center

ChiCTR2000029206 / Suspended临床4期IIT

A real-world study for the efficacy and safety of Monoammonium Glycyrrhizinate and Cysteine and Sodium Chloride Injection in patients with perioperative liver injury

开始日期2020-01-06

申办/合作机构

中山大学

PACTR202401759325189 / Completed临床4期

Effect of Myo-Inositol-in combination with alpha-lipoic acid and cysteine (Celine) based co-treatment on Oocyte Quality in Women with Polycystic Ovarian Syndrome Undergoing Assisted Reproductive Technology

开始日期2019-01-02

申办/合作机构-

100 项与盐酸半胱氨酸相关的临床结果

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100 项与盐酸半胱氨酸相关的转化医学

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100 项与盐酸半胱氨酸相关的专利（医药）

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6,640

项与盐酸半胱氨酸相关的文献（医药）

2024-12-01·Fly

Ribose-cysteine and levodopa abrogate Parkinsonism via the regulation of neurochemical and redox activities in alpha-synuclein transgenic Drosophila melanogaster models.

Article

作者: Olumayowa K Idowu ; Ademola A Oremosu ; Olufunke O Dosumu ; Abdullahi A Mohammed

Parkinson's disease (PD), the most prevalent type of parkinsonism, is a progressive neurodegenerative condition marked by several non-motor and motor symptoms. PD is thought to have a complex aetiology that includes a combination of age, genetic predisposition, and environmental factors. Increased expression of α-synuclein (α-Syn) protein is central to the evolvement of neuropathology in this devastating disorder, but the potential of ribose-cysteine and levodopa in abating pathophysiologic changes in PD model is unknown. Crosses were set up between flies conditionally expressing a pathological variant of human α-Syn (UAS-α-Syn) and those expressing GAL4 in neurons (elav-GAL4) to generate offspring referred to as PD flies. Flies were randomly assigned to five groups (n = 40) from the total population of flies, with each group having five replicates. Groups of PD flies were treated with either 500 mg/kg ribose-cysteine diet, 250 mg/kg levodopa diet, or a combination of the two compounds for 21 days, whereas the control group (w¹¹¹⁸) and the PD group were exposed to a diet without ribose-cysteine or levodopa. In addition to various biochemical and neurochemical assays, longevity, larval motility, and gravitaxis assays were carried out. Locomotive capability, lifespan, fecundity, antioxidant state, and neurotransmitter systems were all significantly (p < 0.05) compromised by overexpression of α-Syn. However, flies treated both ribose cysteine and levodopa showed an overall marked improvement in motor functions, lifespan, fecundity, antioxidant status, and neurotransmitter system functions. In conclusion, ribose-cysteine and levodopa, both singly and in combination, potentiated a therapeutic effect on alpha-synuclein transgenic Drosophila melanogaster models of Parkinsonism.

2024-03-01·Journal of food science and technology

Copper based metal organic framework decorated with gold nanoparticles as a new electrochemical sensor material for the detection of L-Cysteine in milk samples

Article

作者: Perk, Benay ; Tepeli Buyuksunetci, Yudum ; Anik, Ulku

A facile electrochemical sensor based on carbon felt electrode (CFE) modified with gold nanoparticles decorated copper based metal organic framework (AuNPs@Cu-MOF) was achieved for the electrochemical sensing of L-Cysteine (L-Cys). For this purpose, AuNPs@Cu-MOF was synthesized and characterized. The electrochemical behaviors of L-Cys at plain and modified CFEs were investigated via cyclic voltammetry (CV). According CV results, AuNPs@Cu-MOF structure showed a catalytic effect on the oxidation of L-Cys as well as increasing the active electrode surface area by 206% compared to bare CFE. In addition, the pH effect on electrochemical determination of L-Cys at AuNPs@Cu-MOF/CFE was widely examined, and it was determined that the best oxidation peak current of L-Cys was obtained in pH 5 acetate buffer. Moreover, a linear detection range of 30-400 µM for L-Cys with a limit of detection value of 2.21 µM (n = 3) was achieved with the proposed electrochemical sensor. The developed L-Cys sensor was also applied for L-Cys detection in various milk samples and acceptable recovery values were obtained ranging from 100.05 to 108.45%.

Supplementary Information:

The online version contains supplementary material available at 10.1007/s13197-023-05866-1.

2024-02-22·Journal of the science of food and agriculture

The volatile organic compounds generated from the Maillard reaction between L-ascorbic acid and L-cysteine in hot compressed water.

Article

作者: Liang Feng ; Yan Yang ; Ya-Ting Xie ; Wen-Yi Yan ; Ying-Ke Ma ; Sheng Hu ; Ai-Nong Yu

BACKGROUND:

HCW, also known as subcritical water (SCW), refers to high-temperature compressed water in a special physical and chemical state. It is an emerging technology for natural product extraction. The volatile organic compounds (VOCs) generated from the Maillard reaction between L-ascorbic acid (ASA) and L-cysteine (Cys) have attracted significant interest in the flavor and fragrance industry. This study aimed to explore the formation mechanism of VOCs from ASA and Cys and examine the effects of reaction parameters such as temperature, time, and pH in HCW.

RESULTS:

The identified VOCs were predominantly thiophene derivatives, polysulfides, and pyrazine derivatives in HCW. The findings indicated that thiophene derivatives were formed under various pH conditions, with polysulfide formation favored under acidic conditions and pyrazine derivative formation preferred under weak alkaline conditions, specifically at pH 8.0.

CONCLUSION:

The Maillard reaction between ASA and Cys mainly produced thiophenes derivatives, poly-sulfides, and pyrazines derivatives in HCW. The generation mechanism significantly was significantly depends on the surrounding pH condition. This article is protected by copyright. All rights reserved.

项与盐酸半胱氨酸相关的新闻（医药）

2024-03-22

·药精通Bio

读懂此图，教你培养完美肿瘤类器官（附六大肿瘤类器官培养 protocol）

深度聚焦类器官与3D培养论坛，OTC2024论坛合作详询：王晨 180 1628 8769近期，国人团队发表重要综述 Organoids: opportunities and challenges of cancer therapy，仅需一张图简明扼要地总结了不同肿瘤类器官培养方法的关键。从肿瘤组织中通过机械或酶解获得肿瘤细胞，用几乎相同的基础培养基进行培养，而细胞因子是肿瘤类器官培养成功的决定因素之一。不同肿瘤类器官培养方法的简要概述（源自参考文献：Organoids opportunities and challenges of cancer therapy）接下来，我们将重点介绍几种重要的肿瘤类器官培养基组分及方案。1. 胃癌类器官通过构建源自胃癌患者的类器官（PDO），能够直接对临床样本进行研究。Vlachogiannis 等人为了成功培养胃癌类器官，除了使用 ADMEM/F12 培养基和基底膜基质外，还添加了 EGF、Noggin、R-Spondin 1、FGF-10 等细胞因子。胃癌患者的类器官（PDO）培养步骤：如下实验流程和数据均引用Vlachogiannis等人发表的文章（doi: 10.1126/science.aao2774），供参考。1，配置 PDO 培养基：改良的 DMEM/F12 基础培养基，添加 1x B27 additive、1x N2 additive、0.01% BSA、2 mM L-Glutamine、100 units/ml 青霉素-链霉素，以及如下表的细胞因子或其他添加物：*HGF 仅用于胆管癌类器官。2，样本收集：通过图像引导或内窥镜手术收集活检样品。收获后，样本置于冷 PBS 中，并在冰上运输到实验室，20 分钟～2 小时内将组织切片切碎。3，样本处理：组织切碎后，加入 5 ml 5 mM 的 PBS/EDTA，室温静置 15 分钟。用含有 2x TrypLe 的 1 mM PBS/EDTA 5 ml 在 37 ℃ 消化 1 小时。施加机械力（如吹液）以促进消化。4，接种细胞：用改良的 DMEM/F12 培养基收集分离的细胞，并在 4 ℃，1,200 转/分离心 5 分钟，使细胞成球状，用 120 µl GFR 基质胶重悬，接种到 24 孔或 48 孔的平底细胞培养板中。5，将基质在 37 ℃、5% CO2 的细胞培养箱中培养 20 分钟，进行固化。然后添加 500 µl 配置好的 PDO 培养基，要完全覆盖培养孔。6，每两天换一次培养基。2. 结直肠癌类器官患者来源的肿瘤类器官已成为结直肠癌临床前研究的一种出色的模型，原代上皮细胞可在特定生长因子的控制下和 3D 细胞外基质（ECM）中成功培养类器官。肠上皮细胞的自我更新由位于隐窝中的 Lgr5 干细胞驱动。Wnt 信号是维持隐窝干细胞活跃所必需的，Wnt 激动剂 R-Spondin 1 是 Lgr5 的配体，诱导隐窝增生。EGF 信号与肠道增殖有关，Noggin 诱导隐窝数量的急剧增加。因此 van de Wetering 等人开发的结直肠癌类器官培养体系如下：（如下实验流程和数据均引用 van de Wetering 等人发表的文章（doi: 10.1016/j.cell.2015.03.053），供参考。）1. 基础培养基：改良的 DMEM/F12 培养基，添加青霉素-链霉素、10 mM HEPES 和 Glutamax。2. 人肠干细胞培养基：Wnt 条件培养基、R-Spondin 1 条件培养基、Noggin 条件培养基和基础培养基按照 50%、20%、10% 和 20% 的体积比混合，然后加入 1x B27、1.25 mM n-Acetyl Cysteine、10 mM Nicotinamide、50 ng/ml 人 EGF、10 nM Gastrin、500 nM A83-01、3 uM SB202190、10 nM Prostaglandine E2 和 100 mg/ml Primocin。3. H&E 染色分析：通过 H&E 染色对培养后的原代肿瘤细胞和类器官进行分析，发现大多数类器官的特征是存在一个或多个管腔，与原代肿瘤的管状结构相似（如 P8 和 P19b），没有管腔的原代肿瘤会形成没有管腔的紧凑型类器官（P19a）。肿瘤类器官与原代肿瘤上皮相似3. 肝癌类器官肿瘤类器官的培养非常复杂，如肝癌类器。原发性肝细胞癌主要分为肝细胞癌（HCC）、胆管细胞癌（CC）和混合型（CHC）。在肝细胞癌类器官培养中需要使用地塞米松但是胆管细胞性肝细胞癌类器官培养则需加入 R-spondin 1 来维持细胞生长。Broutier 等人发现原发性肝癌（PLC）组织培养获得的类器官可以在体外真实地模拟人类 PLC 的遗传复杂性。构建的类器官包含 PLC 常见的三种亚型：HCC、CC 和 CHC，再现亲本肿瘤的组织结构和表达谱，同时保留患者及亚型之间的差异性，有利于肝癌相关基因和潜在生物标志物的鉴定。肝癌类器官分类培养基：如下实验流程和数据均引用 Broutier 等人发表的文章（doi: 10.1038/nm.4438），供参考。1，基础培养基：改良的 DMEM/F12 培养基，添加 1% 青霉素-链霉素、1% Glutamax、10 mM HEPES 和 1:50 B27 添加剂。2，经典人肝类器官分离培养基：Wnt 条件培养基、R-Spondin 1 条件培养基和基础培养基按照 30%、10% 和 70% 的体积比混合，然后加入 1:100 N2 添加剂、1.25 mM n-Acetyl Cysteine、10 mM Nicotinamide、10 nM 重组人 Gastrin I、50 ng/ml 重组人EGF、100 ng/ml 重组人 FGF10、25 ng/ml 重组人 HGF、25 ng/ml Noggin、5 uM A83-01、10 uM Forskolin、10 uM Y27632。3，肿瘤细胞特异性分离培养基：经典人肝类器官分离培养基中添加 3 mM 地塞米松。4，肝癌类器官培养基：基础培养基、10% （体积比）R-Spondin 1 条件培养基、1:100 N2 添加剂、1.25 mM n-Acetyl Cysteine、10 mM Nicotinamide、10 nM 重组人 Gastrin I、50 ng/ml 重组人 EGF、100 ng/ml 重组人 FGF10、25 ng/ml 重组人 HGF、5 uM A83-01、10 uM Forskolin。4. 乳腺癌类器官乳腺癌类器官的培养基与其他肿瘤的略有不同。Seino 等人在类器官培养基优化过程中发现，Neuregulin 1 与乳腺发育和肿瘤发生有关，将其添加到培养基中可维持类器官有效再生和长期增殖（超过 20 代）。Wnt-3A 对类器官培养条件改善不明显。EGF 对乳腺癌类器官培养具有双重作用，高于 5 ng/ml 促进细胞增殖，但会导致类器官解体，失去自组织功能，低浓度则相反。Hans Clevers 等构建乳腺癌类器官的培养基组分：（如下实验流程和数据均引用 Hans Clevers 等人发表的文章（ doi 10.1016/j.cell.2017.11.010 ），供参考。）a：类器官出现解体时减少或不添加，b：浓度超过 1 μM 会降低类器官生成效率5. 肺癌类器官培养肺癌是全球常见的恶性实体肿瘤，但是肺癌早期临床表现不明显，容易与其他感染混淆而被忽视，一般确诊时已发展到中晚期，因此，及时准确地筛选合适的抗肿瘤药物对肺癌患者非常重要。1 份从原发性肿瘤组织中衍生出肺腺癌类器官（成功率达到 80%）的培养基组分：（如下实验流程和数据均引用Zhichao Li等人发表的文章（10.1016/j.xpro.2020.100239 ），供参考。）试剂详细配制指南关键：所有操作都应在二级生物安全柜中进行，以保持无菌状态。1200 μg/mL FGF-10 (10,000 X)a. 取100 μg溶于0.5 mL 无菌的 PBS（加 0.1% BSA，质量/体积）。b. 每管 5 μL，将溶液分装到 1.5 mL 的离心管中，可在 -20 ℃ 保持 3 个月。2250 μg/mL FGF-7 (12,500 X)a. 取 100 μg 溶于 0.4 mL 无菌的 PBS（加 0.1% BSA，质量/体积）。b. 每管 4 μL，将溶液分装到 1.5 mL 的离心管中，可在 -20 ℃ 保持 3 个月。3100 μg/mL R-spondin 1 (200 X)a. 取 250 μg 溶于 2.5 mL 无菌的 PBS（加 0.1% BSA，质量/体积）。b. 每管 250 μL，将溶液分装到 1.5 mL 的离心管中，可在 -20 ℃ 保持 1 个月。4100 μg/mL Noggin (1,000 X)a. 取 100 μg 溶于 1 mL 无菌的 PBS（加 0.1% BSA，质量/体积）。b. 每管 50 μL，将溶液分装到 1.5 mL 的离心管中，可在 -20 ℃ 保持 2 个月。5100 mM Y-27632 dihydrochloride (10,000 X)a. 取 50 mg 溶于 1.56 mL 的超纯水中。b. 用 0.22 μm 的过滤器过滤溶液。c. 每管 5 μL，将溶液分装到 1.5 mL 的离心管中，可在 -20 ℃ 保持 6 个月。6500 mM N-Acetylcysteine (400 X)a. 取 408 mg 溶于超纯水，至终体积为 5 mL。b. 用 0.22 μm 的过滤器过滤溶液。c. 每管 125 μL，将溶液分装到 1.5 mL 的离心管中，可在 -20 ℃ 保持 3 个月。72 M Nicotinamide (200 X)a. 取 6.1 g 溶于 1x PBS，至终体积为 25 mL。b. 用 0.22 μm 的过滤器过滤溶液。c. 每管 250 μL，将溶液分装到 1.5 mL 的离心管中，可在 -20 ℃ 保持 3 个月。830 mM SB202190 单盐酸盐水合物 (3,000 X)a. 取 5 mg 溶于 0.453 mL 的二甲基亚砜（DMSO）中b. 每管 16.67 μL，将溶液分装到 1.5 mL 的黑色离心管中，可在 -20 ℃ 保持 3 个月。关键：不要用聚偏二氟乙烯（PVDF）膜过滤含有 DMSO 的溶液，因为 DMSO 可以溶解这种膜。关键：使用黑色离心管，防止该溶液受到光照。建议：过滤 DMSO 溶液，可以使用 DMSO 安全过滤器或尼龙膜过滤器。925 mM A83-01 (50,000 X)a. 取 5 mg 溶于 0.474 mL 的二甲基亚砜（DMSO）中b. 每管 2 μL，将溶液分装到 1.5 mL 的离心管中，可在 -20 ℃ 保持 3 个月。10分装基质胶a. 取一瓶新的基质胶，放于 4 ℃ 冰箱中解冻 8 小时。b. 将瓶子倒置几次，使其充分混合。c. 取 1.5 mL 的离心管置于冰上冷却。d. 将基质胶和 1.5 mL 的离心管置于冰上，按实验所需体积将基质胶分装到 1.5 mL 的离心管中，并在 -20 ℃ 保存。关键：移液器枪头应在抽吸基质胶之前，为了防止基质胶黏在枪头上，应通过多次抽吸和排出冷的无菌 PBS（含 01.% BSA）进行预湿和预冷。注意：基质胶是一种重组基膜制剂，富含胞外蛋白，应避免反复冻融。6. 卵巢癌类器官目前，培养卵巢癌类器官的方法有多种，但成功率较高的是使用基质胶和改良 DMEM/F12 基础培养基的组合，并辅以 EGF、FGF2、FGF10、Noggin、Rspondin1、p38 抑制剂等，可使类器官培养成功率达到 80%～90%。Senkowski 等人为了获得高级别浆液性卵巢癌（HGSC）类器官的长期培养方案，对培养基添加组分进行优化，获得两种成功率更高的配方。并且体外培养的 HGSC 类器官保留了原始肿瘤的基因组和表型特征，其药物反应与临床结果具有相关性。培养基优化主要涉及添加物种类和浓度，尤其是细胞因子类。研究人员在改良型 DMEM/F12 培养基中加入 Glutamax、Primocin、N-acetyl-cysteine 和 B27 添加剂构成基础培养基。通过分析添加剂对 HGSC 原代细胞短期类器官形成和生长的影响，使用添加 FGF-10、SB202190 和 A83-10 的基础培养基（M 0.1）进行优化。但 M 0.1 培养基能促进生长，但在传代后并不能维持生长。进一步确定 HGSC 类器官培养基配方中加入了 5 mM 烟酰胺，形成培养基 M1。在培养基 M1 中添加 EGF、Heregulin-β-1、Hydrocortisone 和 Forskolin，形成培养基 M2。对于不同的样品，M2 可能具有促进或抑制类器官生长的「矛盾」结果，因此为了最大限度地提高类器官形成，每个 HGSC 样品应在两种不同的培养基 M1 和 M2 中平行培养。类器官形成和长期培养测试的主要因子及其影响概述7. 其他肿瘤类器官膀胱癌类器官利用 CTOS 培养基进行培养，主要成分为改良型 DMEM/F12、B27、A83-01，需补充添加 FGF10、FGF7、HER3。培养前列腺类器官时，一般会加入 FGF10、FGF2、p38 抑制剂 SB202190、Rho 激酶抑制剂等，而小鼠前列腺癌类器官无需添加 FGF10、FGF2、PGE2、烟酰胺和 SB202190。文章来源：生物学霸END深度聚焦类器官与3D培养论坛，OTC2024论坛合作详询：王晨 180 1628 8769戳“阅读原文”立即领取限量免费参会名额!

临床研究

2024-02-09

·药渡

抗体偶联药物（ADC）定量生物分析及自动化助力药代动力学评估

前言抗体偶联药物（Antibody-Drug Conjugates，ADC）是结合了单克隆抗体的高度靶向性和小分子毒素的强细胞毒性的新疗法，由于其精准而强大的作用机理，已成为抗肿瘤抗体药物开发研究的新热点和重要趋势[1]。ADC药物能够有效地杀伤癌细胞，同时不伤害正常组织，从而提高抗癌治疗的治疗指数，被誉为“精确制导的魔法子弹”。ADC由三个部分组成 (图1)[2]，包括靶向性的抗体部分、发挥最主要功能的小分子载荷以及连接两者的连接子。ADC中的单克隆抗体通过与肿瘤细胞上过度表达的靶抗原结合后，ADC被内化到肿瘤细胞中，由于溶酶体蛋白酶的存在，细胞毒性有效载荷（Payload）与抗体部分分离，细胞毒性载荷启动细胞损伤或凋亡以杀死癌细胞[3]。如有效载荷可以扩散到细胞外基质，杀死邻近癌细胞，称之为旁观者效应（Bystander Effect），这进一步增强了ADC的功效。全面地了解ADC药物的药代动力学特征，对于ADC药物的设计和开发至关重要，本文将重点介绍ADC药物的药代动力学评估生物分析策略和ADC药物定量生物分析平台，为药物研发者提供思路。图1. ADC药物的结构[2]ADC药物和传统的抗体药物或小分子药物相比，具有结构复杂的特点，多种因素会影响其疗效，同时ADC药物体内动态过程多样，从分布、解偶联、代谢和消除评价ADC药物的暴露-功效和暴露-安全关系，对于药物设计和后续开发均至关重要[4]。ADC药物在各种组织中的吸收、分布、代谢和排泄（Absorption，Distribution，Metabolism，Excretion，ADME）直接影响ADC的疗效和安全性，相比其他药物更为复杂，也更具挑战[5]。ADC药物不仅需要分析大分子（总抗体及偶联抗体）还需要分析小分子（游离载荷及偶联载荷）成分，ADC药物DMPK研究策略（图2）包括体外稳定性和payload释放实验、体内PK/PD研究等等。药明康德DMPK可以根据客户项目的需求，提出个性化的ADC药物药代动力学研究策略和具体的实验方案，加速药物的研发和注册申报。图2. ADC药物DMPK研究策略评估总抗体（Total Antibody）、偶联抗体 (Conjugated Antibody)、游离载荷（Free Payload）和相关代谢物的定量生物分析贯穿ADC药物早期发现/筛选、临床前和临床的各个阶段。为了正确评估ADC药物的ADME特性，需要综合性的生物分析手段。在ADC药物生物分析中，配体结合分析（Ligand Binding Assay，LBA）是最常用的生物分析方法。LBA作为大分子定量研究领域的“金标准”，具有高灵敏度和高通量等优势，常用于检测总抗体、偶联抗体。分析难点在于需要针对不同的种属和基质类型进行关键试剂优化，因此LBA方法开发周期长，同时无法提供ADC的结构信息和相关的生物转化信息。如若无法获得关键试剂，可以使用替代方法进行检测。随着质谱技术的不断发展，液相色谱-串联高分辨质谱(Liquid Chromatography-High Resolution Mass Spectrometry，LC-HRMS) 技术正越来越多地被用于大分子表征和定量，作为替代方法，LC-HRMS主要用于总抗体和偶联抗体的测定，具有时间周期短，通用性好等优点。关于不同生物分析平台详细介绍，欢迎参考我们此前发布的文章：整合性生物分析在抗体偶联药物（ADC）DMPK研究中的应用。下文根据分析物种类的不同，分别介绍对应的定量生物分析方法。1游离载荷（Free Payload）LC-MS/MS定量分析影响ADC毒性很大因素在于游离载荷，因为游离有效载荷可能介导非预期的脱靶毒性，因此监测游离载荷十分必要，这也成为ADC研究中非常重要的一部分。游离载荷在血浆中浓度非常低，对灵敏度提出了更大的挑战，我们使用灵敏度较高的AB 6500、AB 7500进行Free Payload的定量分析。图3为20 pg/mL MMAE在大鼠血浆样品中的LC-MS/MS色谱图，定量低限仍有下降空间，线性满足方法要求。游离载荷的LC-MS/MS定量分析经常应用于判断ADC药物储存过程中是否释放Free Payload、抗体和毒素的连接子血浆稳定性、毒素本身生物基质中稳定性、小分子PK和组织分布等实验中。图3. 20 pg/mL MMAE大鼠血浆样品中的LC-MS/MS色谱图和标准曲线2总抗体（Total Antibody）和偶联位点分析总抗体通常使用LBA平台进行分析。LBA分为特异性方法（Specific Method）或者通用性方法（Generic Method），通常会准备至少两套关键试剂，确保方法开发顺利完成。同时在方法开发中需要考察方法对于完整ADC药物和裸抗的信号响应是否符合一般药代动力学的规律，也应该通过桥接实验考察裸抗与ADC信号的一致性。关于LBA平台详细策略，欢迎参考我们此前发布的文章：基于LBA技术的抗体类药物临床前PK生物分析策略与路径。总抗体分析也可以使用特征性肽段（Surrogate Peptide）方法进行测定，如图4所示，为了减少来自基质的干扰，使用免疫亲和磁珠对ADC进行富集，再经胰蛋白酶酶解后，生成特征多肽，利用LC-MS/MS进行定量分析。该方法灵敏度高，特异性强。目前项目组可以根据抗体序列利用软件进行特征肽段方法开发及质谱方法优化，同时利用肽图可以对偶联位点进行监测。目前该方法已经成功用于体内PK实验和体外血浆稳定性实验（如图5所示），方法的定量低限在ng级别同时线性符合方法学要求。（左右滑动，查看更多）图4. Surrogate Peptide定量流程图5. 不同种属特征肽段定量浓度-时间分布及定量低限和标准曲线图3偶联抗体（Conjugated ADC）分析偶联抗体通常使用LBA平台进行分析，利用抗载荷抗体（Anti Payload Antibody）进行捕获，目前我们针对热门的载荷均有试剂备库，同时已经建立in house方法有MMAE、MMAF、DM1/DM4、SN-39和DXd等，满足快速分析的需求。如果无法获得抗载荷抗体，可以使用替代方法来进行分析偶联抗体。LC-HRMS方法通过借助对ADC抗体部分具有特异性结合的抗体或抗原，从生物基质中将目标物捕获出来，如图6所示，LC-HRMS方法在完整（Intact）或亚基（Subunit）水平分析ADC药物，通过计算得出偶联抗体的值。图6. 不同偶联方式ADC药物的LC-HRMS分析流程4DAR值分析药物抗体比(Drug-Antibody-Ratio， DAR)表示每个ADC药物抗体部分偶联载荷的数量，DAR值随着药物代谢不断变化，是描述ADC药物安全性和有效性的重要参数。疏水相互作用色谱（Hydrophobic Interaction Chromatography，HIC）被认为是半胱氨酸偶联ADC药物DAR值分析的金标准[6]，但该方法对ADC需求量大（20-50 μg）且需要样品中有较高的ADC浓度[7]。近年来，LC-HRMS以其独特的优势，在Intact和Subunit水平分析ADC药物DAR值提供了一种可行的方法[8]，与HIC-UV法相比，LC-HRMS法所需供试品更少，可以实现不同DAR值ADC的表征。关于DAR分析详细策略，欢迎参考我们此前发布的文章：基于高分辨质谱技术的完整蛋白定量定性一体化生物分析。综上所述，测试不同的分析物需要选择不同的平台或方法，如表1所示，每种平台或方法都有其优势及其适用场景，针对客户不同需求，提出适合的实验方案。表1. ADC药物分析平台选择自动化-提升ADC免疫捕获效率和精度从复杂的生物基质中提取目标ADC药物通常利用磁珠进行免疫亲和捕获，但是该方法关键控制点多、依赖人员经验，存在实验流程长、数据重现性差等缺点，目前药明康德药性评价部Non-GLP生物分析团队采用微色谱（IMCSTIPS）与自动化结合，将免疫亲和捕获时间从手动8 h缩短到自动2 h，同时提升数据的精度和交付的质量。如图7所示，我们利用通用型和特异性的捕获试剂，同时尝试了常规血清和IgG deplete血清分别进行自动化免疫捕获效率的测试，并利用LC-HRMS进行检测。从图7中可以看出，如利用特异性的捕获试剂在常规血清中捕获效率80%左右；如果使用IgG deplete血清和特异性的捕获试剂，捕获效率可达90%。图7. 自动化-免疫亲和捕获效率图团队开发并验证了自动化-免疫亲和捕获用于单抗类药物和ADC类药物的PK分析能力。以DS-8201为例，如图8所示，利用Biotinylated-Her2和IMCS进行免疫亲和捕获，线性范围在1-50 µg/mL呈现良好的线性回归。图8. 大鼠基质中利用自动化-免疫亲和捕获DS-8201轻链和重链线性图自动化数据捕捉-提升数据处理效率和精度ADC在LC-HRMS产生的海量数据，手动收集数据时间长，且易错不准确，药明康德DMPK利用自编代码进行自动数据整理，降低数据收集的时间，从手动整理数据8 h降低至自动化5 min，提升数据抓取效率和准确度。结语ADC药物结构复杂，体内过程多样，ADC药物的稳定性、安全性和有效性需要多种生物学分析平台共同助力完成，以观察ADC的独特生物转化和获得丰富、准确的数据。随着更多创新的偶联策略、抗体支架和新型弹头被用于下一代ADC，生物分析技术的未来发展将需要解决这些日益复杂的问题。参考文献：[1] Yu J, Wu S, Li R, et al. Novel ADCs and combination therapy in urothelial carcinoma: latest updates from the 2023 ASCO-GU Cancers Symposium[J].Journal of Hematology & Oncology, 2023, 16(1).[2] Fu Z, Li S, Han S, et al. Antibody drug conjugate: the "biological missile" for targeted cancer therapy [J]. Sig Transduct Target Ther , 2022(004):007.[3] Samantasinghar A, Sunildutt N P, Ahmed F, et al. A comprehensive review of key factors affecting the efficacy of antibody drug conjugate[J]. Biomedicine & Pharmacotherapy, 2023(161): 114408.[4] Mou S, Huang Y, Rosenbaum A. ADME Considerations and Bioanalytical Strategies for Pharmacokinetic Assessments of Antibody-Drug Conjugates[J]. Antibodies, 2018, 7(4).[5] Kraynov E, Kamath A V, Walles M, et al. Current Approaches for Absorption, Distribution, Metabolism, and Excretion Characterization of Antibody-Drug Conjugates: An Industry White Paper. Drug Metab. Dispos. (2016) 44, 617-623.[6] Ouyang J. Drug-to-antibody ratio (DAR) and drug load distribution by hydrophobic interaction chromatography and reversed phase high-performance liquid chromatography[J]. Methods in Molecular Biology, 2013, 1045:275.[7] Li K, Zhang Z, Lin Z J, et al. Accurate determination of drug-to-antibody ratio of interchain cysteine-linked antibody–drug conjugates by LC-HRMS[J]. Analytical and Bioanalytical Chemistry, 2019, 412(9):1-8.[8] Martelet A, Garrigue V, Zhang Z, et al. Multi-attribute method based characterization of antibody drug conjugates (ADC) at the intact and subunit levels[J]. Journal of pharmaceutical and biomedical analysis, 2021, 201:114094. （下滑查看更多）作者：曲栗，李陟昱，邢丽丽编辑：方健，钱卉娟设计：倪德伟，张莹莹免责声明“药渡”公众号所转载该篇文章来源于其他公众号平台，主要目的在于分享行业相关知识，传递当前最新资讯。图片、文章版权均属于原作者所有，如有侵权，请及时告知，我们会在24小时内删除相关信息。微信公众号的推送规则又双叒叕改啦，如果您不点个“在看”或者没设为"星标"，我们可能就消散在茫茫文海之中~点这里，千万不要错过药渡的最新消息哦！👇👇

抗体药物偶联物

2024-02-04

·生物制品圈

抗体人春节充电必备|ADC药物书籍：研发、生产、注册、临床及市场

抗体偶联药物（ADC）是一类意义非凡的高效生物药，有望成为广泛应用的癌症治疗方法。ADC是由单克隆抗体与细胞毒性药物通过生物活性连接子偶联而成的化合物分子。与化疗不同，ADC的研究理念是仅标记并杀死癌细胞，因而成为靶向性较高的治疗方法。ADC在癌症领域发挥着举足轻重的作用，全球市场上约有12种ADC用于血液瘤和恶性实体瘤。辉瑞公司、基因泰克公司、吉利德科学公司以及许多其他药企都在大力投资ADC，市场竞争处于白热化状态。作为靶向肿瘤的生物治疗药物，ADC的治疗潜力非比寻常；ADC只有在内化到肿瘤细胞后才会启动其作用机制，然后释放细胞毒性有效载荷，从而选择性地杀死恶性肿瘤细胞。ADC无疑是近几年最火的领域之一，2022年全球销售规模超过70亿美元，去年前九个月主要产品销售额已超过70亿美元，全年有望突破百亿美元大关，目前全球已经有15个ADC获批上市，在中国获批的有7个。目前比较热门的靶点有CD22、HER2、EGFR、Trop-2、BCMA、Nectin-4等，常见适应症主要包括乳腺癌、肺癌、胃癌和血癌等肿瘤。ADC赛道席卷全球，在研的ADC药物多达几百款，国内外药企不惜重金纷纷布局ADC，如辉瑞斥资430亿美元收购Seagen，近期默沙东以高达220亿美元与第一三共达成合成协议，共同开发第一三共的三款ADC产品。国内也有通过自研、并购或License in等模式布局ADC管线的药企，其中科伦药业的HER2 ADC产品A166正在申报上市，科伦药业的SKB264、恒瑞医药的SHR-A1811、石药的DP303c和乐普生物的MRG002等都处于临床3期。越来越多的ADC的获批上市，截至目前，全球获批上市ADC药物有15个，在中国获批上市的产品有7个。也有多款药物进入临床阶段，其中靶向HER2的ADC研究最火热，其次是Trop-2等。图1. 获批上市的ADC药物（药时代整理）图2. 部分在研的临床3期ADC药物（药时代整理）图3. 部分临床2期在研的ADC药物（药时代整理）图4. 部分临床1期在研的ADC药物（药时代整理）值新书佳节之际，小编精选了ADC药物在研发、生产、注册、临床及市场的8本书籍和大家一起学习交流，有兴趣的老师可以扫码登记，符合要求方可加入我们学习行列。快快加入我们喔，一起过一个充实而又愉快的春节！Antibody-Drug Conjugates and Cellular Metabolic DynamicsThis book summarizes the related research achievements in Antibody-drug conjugates (ADCs) and their cell metabolism kinetics. The book has three main parts. The first part describes the basic theory of ADCs, including the basic concept and structure of ADCs, and the relationship between the targets of ADCs and their specific functions. The second part mainly introduces the endocytosis and intracellular metabolism of ADCs, including the relationship between endocytosis and ADC activity, the endocytosis and intracellular transport of ADCs, the distribution and metabolism of ADC in vivo. Then it discusses the new formats and research technology of ADCs, including the application of miniaturized antibodies in ADC synthesis, novel carriers for ADC design, the technology and application of site-specific conjugation, and approaches for analyzing the drug: antibody ratio (DAR), the study of pharmacokinetics of ADCs. This book combines the basic theory with the research technology. It can be used as a reference book for students, teachers and researchers of biomedical field.Chemical Linkers in Antibody-Drug Conjugates (ADCs) 2021The covalent conjugation of potent cytotoxic agents to monoclonal antibodies, known as antibody-drug conjugates (ADCs) is a powerful approach in the field of targeted treatment of cancer. Clearly, both monoclonal antibody and cytotoxic payload are crucial elements in determining the clinical value of an ADC and have receive ample attention. However, the structural element connecting the two –the chemical linker– also plays an essential role in mode-of-action, efficacy, pharmacokinetics and safety profile of an ADC, but is often underappreciated in considerations of ADC design.Chemical Linkers in Antibody–Drug Conjugatesaims to shine a detailed light on the various key attributes of chemical linkers in ADCs, for drug-to-antibody ratio, for stability, for release mechanism of payload, for pharmacokinetics, for stability determination, and for efficacy and safety. Ideal for postgraduate students and active researchers in drug discovery and development, this book provides a comprehensive description of linkers used in ADCs (clinical and late preclinical), insight into key quality attributes of linkers for ADCs, and aids the reader in understanding the role of linker chemistry and designing new ADCs.Antibody-Drug Conjugates: Methods and ProtocolsThis volume looks at key methodologies that are commonly used across antibody drug conjugates (ADCs) programs. The chapters in this book cover topics such as conjugations to endogenous cysteine residues; click chemistry conjugations; antibody conjugations via glycosyl remodeling; analysis of ADCs by native mass spectrometry; characterization of ADCs by capillary electrophoresis; LC/MS methods for studying lysosomal ADC catabolism; and determination of ADC concentration by ligand-binding assays. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and practical, Antibody-Drug Conjugates: Methods and Protocols is a valuable resource that aims to lower the “activation barrier” when undertaking a new discipline, and provides a “toolbox” for the next generation of ADC scientists.Next Generation Antibody Drug Conjugates (ADCs) and ImmunotoxinsThis book describes the newest developments in antibody drug conjugates and immunotoxins, paving their way to clinical application. Lessons learned from the current state of the art are used to further improve our understanding of their mechanisms of action and off target activities.The book introduces scientists to all of the prerequisites that must be properly addressed, including identification of the right target, specific traits of target binding antibodies, proper selection of the toxic payload, internalization induced by binding, and next generation conjugation and linker technologies. These knowledge-based, revolutionary new drug principles will form the cornerstone of the future standard of care and will lead to major advances in application, as well as improved quality of life and patient survival rates. This book will be of interest to biotech companies and researchers working in the fields of immunology, pharmacology, and oncology.Antibody-Drug Conjugates: Fundamentals, Drug Development, and Clinical Outcomes to Target CancerProviding practical and proven solutions for antibody-drug conjugate (ADC) drug discovery success in oncology, this book helps readers improve the drug safety and therapeutic efficacy of ADCs to kill targeted tumor cells.• Discusses the basics, drug delivery strategies, pharmacology and toxicology, and regulatory approval strategies• Covers the conduct and design of oncology clinical trials and the use of ADCs for tumor imaging• Includes case studies of ADCs in oncology drug development• Features contributions from highly-regarded experts on the frontlines of ADC research and developmentAntibody-Drug Conjugates and Immunotoxins: From Pre-Clinical Development to Therapeutic ApplicationsThis volume gathers the leading research on antibody-drug conjugates and immunotoxins. Following a rigorous overview, the volume delves into focused sections on all aspects of ADCs and ITs from clinical development through to targeted therapeutic applications and the latest technologies.Antibody-Drug Conjugates: The 21st Century Magic Bullets for CancerThis authoritative volume provides a holistic picture of antibody-drug conjugates (ADCs). Fourteen comprehensive chapters are divided into six sections including an introduction to ADCs, the ADC construct, development issues, landscape, IP and pharmacoeconomics, case studies, and the future of the field. The book examines everything from the selection of the antibody, the drug, and the linker to a discussion of developmental issues such as formulations, bio-analysis, pharmacokinetic-pharmacodynamic relationships, and toxicological and regulatory challenges. It also explores pharmacoecomonics and intellectual properties, including recently issued patents and the cost analysis of drug therapy. Case studies are presented for the three ADCs that have received FDA approval: gemtuzumab ozogamicin (Mylotarg®), Brentuximab vedotin (Adcetris®), and ado-trastuzumab emtansine (Kadcyla®), as well as an ADC in late-stage clinical trials, glembatumumab vedotin (CDX-011). Finally, the volume presents a perspective by the editors on the future directions of ADC development and clinical applications.Antibody-Drug Conjugates is a practical and systematic resource for scientists, professors, and students interested in expanding their knowledge of cutting-edge research in this exciting field.Innovations for Next-Generation Antibody-Drug Conjugates Antibody-drug conjugates (ADCs) stand at the verge of a transformation. Scores of clinical programs have yielded only a few regulatory approvals, but a wave of technological innovation now empowers us to overcome past technical challenges. This volume focuses on the next generation of ADCs and the innovations that will enable them. The book inspires the future by integrating the field’s history with novel strategies and cutting-edge technologies. While the book primarily addresses ADCs for solid tumors, the last chapter explores the emerging interest in using ADCs to treat other diseases.The therapeutic rationale of ADCs is strong: to direct small molecules to the desired site of action (and away from normal tissues) by conjugation to antibodies or other targeting moieties. However, the combination of small and large molecules imposes deep complexity to lead optimization, pharmacokinetics, toxicology, analytics and manufacturing. The field has made significant advances in all of these areas by improving target selection, ADC design, manufacturing methods and clinical strategies. These innovations will inspire and educate scientists who are designing next-generation ADCs with the potential to transform the lives of patients.识别微信二维码，添加生物制品圈小编，符合条件者即可加入生物制品微信群！请注明：姓名+研究方向！版权声明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。

抗体药物偶联物引进/卖出临床2期并购临床3期

100 项与盐酸半胱氨酸相关的药物交易

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外链

KEGG	Wiki	ATC	Drug Bank
D02326	盐酸半胱氨酸	A16AA	DB00151

研发状态

适应症	最高研发状态	国家/地区	公司
肿瘤	批准上市	中国更多	陕西兴邦药业有限公司更多
水电解质代谢紊乱	批准上市	美国更多	Exela Pharma Sciences LLC 更多
营养紊乱	批准上市	美国更多	Exela Pharma Sciences LLC 更多
肝病	批准上市	美国更多	Exela Pharma Sciences LLC 更多
膳食补充剂	撤市	美国更多	Baxter Healthcare Corp. 更多

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临床结果

适应症

分期

评价

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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


ARVO2015	N/A	-	-	L-cysteine	積襯壓窪醖衊獵襯壓願(選構醖醖遞艱獵製壓鬱) = 網糧鹹艱築膚餘淵膚鏇築鬱觸鏇範鹹齋廠蓋齋 (鏇醖衊顧蓋衊淵觸醖鑰 )	-	2015-06-01
				Anandamide	積襯壓窪醖衊獵襯壓願(選構醖醖遞艱獵製壓鬱) = 齋繭遞鏇醖簾鏇繭餘繭築鬱觸鏇範鹹齋廠蓋齋 (鏇醖衊顧蓋衊淵觸醖鑰 )
AUA2018	N/A	肾结石	-	Non-calcified stones (struvite, cysteine, uric acid)	繭餘獵艱製齋醖鏇衊蓋(積構繭艱壓願廠選艱積) = 壓簾窪網糧夢觸襯築選膚艱憲壓淵艱襯壓鏇齋 (艱淵積醖積簾鏇鏇鏇鏇 )	-	2018-04-01
EULAR2010	N/A	L-cysteine \| cystathione-γ-lyase (CSE) \| cystathione-β-synthase (CBS)	-	GYY4137	窪獵鬱糧艱獵顧網選壓(壓襯繭鑰夢廠築築選繭) = GYY4137 also inhibited TNFα (1ng/ml) induced expression of iNOS and COX-2 膚襯獵糧網願網顧鏇築 (餘廠壓獵壓築積鑰製範 ) 更多	-	2010-06-16
				L-cysteine
ARVO2011	N/A	视网膜疾患 CD34+	-	Cysteine-Rich Protein 61 (CCN1/Cyr61)	(願廠鏇簾顧積獵衊構範) = 襯鬱鏇簾糧醖鏇獵鹹獵鹹憲鏇鏇遞齋糧觸獵窪 (簾積蓋繭夢積鏇壓鏇製 )	-	2011-04-01
				Cysteine-Rich Protein 61 (CCN1/Cyr61)	(願廠鏇簾顧積獵衊構範) = 夢窪艱淵範壓築衊憲鑰鹹憲鏇鏇遞齋糧觸獵窪 (簾積蓋繭夢積鏇壓鏇製 )