A Specific Mechanism for non-specific inhibition

非特异性抑制的特异性机制

• 批准号: 9182837
• 负责人: Brian K Shoichet
• 金额: $ 31.77万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9182837
• 关键词: Adsorption; Animals; Antibodies; Antibody-drug conjugates; Attention; Behavior; Binding; Biological; Biology; Cell Culture Techniques; Cell Nucleus; Cells; Cellular Assay; Characteristics; Chemicals; Clinical; Clinical Trials; Colloids; Deposition; Drug Delivery Systems; Drug Formulations; Drug Kinetics; ERBB2 gene; Environment; Feasibility Studies; Fluorescence; Fulvestrant; Intestines; Itraconazole; Libraries; Liquid substance; Membrane; Membrane Proteins; Modeling; Mono-S; Morphologic artifacts; Pharmaceutical Preparations; Physical Chemistry; Plasma; Play; Property; Proteins; Reagent; Research; Role; Specificity; Staging; Staining method; Stains; Structure; Techniques; Testing; abstracting; antibody conjugate; drug discovery; drug distribution; experience; improved; in vivo; mouse model; particle; programs; protein aggregate; screening; small molecule; tumor

Abstract/ Project Summary Many organic molecules aggregate at micromolar and sub-micromolar concentrations; these colloidal aggregates are the dominant artifact in early drug discovery and chemical biology. The colloids form a state intermediate between true solution and precipitate, with properties that differ from either. They inhibit both soluble and membrane proteins by direct binding followed by partial denaturation. These are the conclusions of the first decade of this program of research. There is no modern screening group that is unaware of colloidal aggregation, and all use the techniques developed by this project to detect them. A surprise of the last period was the range of conditions in which the colloids were stable, from cell culture, to plasma, to simulated intestinal fluid. In the next period, we investigate colloidal aggregates in cellular assays, drug repurposing, drug formulation, and animal pharmacokinetics. More ambitiously still, we attempt to optimize these unusual particles for stability and antibody loading, for delivery. The specific aims are: 1. The impact and mechanism of colloidal aggregation at increasing levels of biological complexity. We investigate a. the persistence of colloid aggregation among late stage clinical candidates, collaborating with Novartis. b. Similarly, we test for aggregation of about 100 likely-to-aggregate drugs in repurposing libraries. Experience with itraconazole suggests that repurposing screens can mislead all the way to clinical trials, if uncontrolled for mechanism. c. We continue to investigate the structure and mechanism of aggregates, using SAXS to explore whether they are hollow or filled, whether they adsorb or absorb proteins, whether they are micro-crystaline or fluid, and their characteristic inhibitory incubation effects. Finally, we investigate d. The effect of aggregation on whole animal pharmacokinetics. 2. Exploiting of colloidal aggregates for protein delivery. An unexpected observation from the last period was that colloidal aggregates could be co-formulated for stability, homogeneity, and protein loading. Exploiting this observation, we investigate a. Colloidal drug antibody conjugates that specifically deliver drug payloads to cells. In these particles, the delivery vehicle—the colloid—is almost 100% drug. Cell delivery deposits 107 drug molecules per colloid. b. optimization of co-formulated colloids for stability, protein loading, size, and mono-dispersity, each key to pragmatic use of the particles. Finally, we c. Investigate these colloid antibody conjugates as vehicles to deliver large drug payloads to tumors in a mouse model. Extensive preliminary results support the feasibility of these studies.

摘要/项目摘要 许多有机分子在微摩尔和亚微摩尔浓度下聚集；这些胶体 聚集体是早期药物发现和化学生物学中的主要伪影。胶体形成状态 在真实溶液和沉淀之间中间，与两个不同的特性。他们抑制两者 通过直接结合，然后进行部分变性，可溶性和膜蛋白。这些是结论 该研究计划的头十年。没有现代筛选小组不知道 胶体聚集，所有人都使用该项目开发的技术来检测它们。 最后一个时期的一个惊喜是，从细胞培养到稳定的气体稳定的条件范围 等离子体，用于模拟肠液。在下一个时期，我们研究了细胞测定中的胶体聚集体， 药物重新利用，药物配方和动物药代动力学。更加雄心勃勃，我们试图 优化这些异常颗粒的稳定性和抗体加载，以递送。具体目的是： 1。胶体聚集在生物学水平升高时的影响和机制 复杂。我们调查一个。晚期临床候选者中结石聚集的持续性， 与诺华合作。 b。同样，我们测试了大约100种可能聚集的药物的聚集 重新利用库。 Itraconazole的经验表明，重新利用屏幕可能会一直误导 进行临床试验，如果无法控制机制。 c。我们继续研究 聚集体，使用SAXS探索它们是空心还是填充，无论它们是吸附还是吸收蛋白质， 它们是微晶还是液体，及其特征性抑制孵育作用。最后，我们 调查d。聚集对整个动物药代动力学的影响。 2。利用胶体聚集体进行蛋白质递送。最后的意外观察 周期是可以将胶体聚集体共同成型，以达到稳定性，同质性和蛋白质负荷。 利用这一观察结果，我们研究了。胶体药物抗体偶联物，专门输送药物 有效载荷给细胞。在这些颗粒中，递送载体（结石）几乎是100％的药物。电池输送 每个胶体沉积107个药物分子。 b。优化协调胶体的稳定性，蛋白质载荷， 大小和单分散性，每个键的颗粒使用键。最后，我们c。研究这些胶体 抗体偶联作为车辆，以在小鼠模型中为肿瘤传递大型药物有效载荷。 广泛的初步结果支持这些研究的可行性。

项目成果

An Aggregation Advisor for Ligand Discovery.

• DOI: 10.1021/acs.jmedchem.5b01105
• 发表时间: 2015-09-10
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Irwin JJ;Duan D;Torosyan H;Doak AK;Ziebart KT;Sterling T;Tumanian G;Shoichet BK
• 通讯作者: Shoichet BK

Colloidal aggregation: from screening nuisance to formulation nuance.

• DOI: 10.1016/j.nantod.2018.02.011
• 发表时间: 2018-04
• 期刊: Nano today
• 影响因子: 17.4
• 作者: Ganesh AN;Donders EN;Shoichet BK;Shoichet MS
• 通讯作者: Shoichet MS

Promiscuous aggregate-based inhibitors promote enzyme unfolding.

• DOI: 10.1021/jm801605r
• 发表时间: 2009-04-09
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Coan KE;Maltby DA;Burlingame AL;Shoichet BK
• 通讯作者: Shoichet BK

Novel cruzain inhibitors for the treatment of Chagas' disease.

• DOI: 10.1111/j.1747-0285.2012.01416.x
• 发表时间: 2012-09
• 期刊: Chemical biology & drug design
• 影响因子: 3
• 作者: Rogers KE;Keränen H;Durrant JD;Ratnam J;Doak A;Arkin MR;McCammon JA
• 通讯作者: McCammon JA

Colloidal drug formulations can explain "bell-shaped" concentration-response curves.

• DOI: 10.1021/cb4007584
• 发表时间: 2014-03-21
• 期刊: ACS CHEMICAL BIOLOGY
• 影响因子: 4
• 作者: Owen, Shawn C.;Doak, Allison K.;Ganesh, Ahil N.;Nedyalkova, Lyudmila;McLaughlin, Christopher K.;Shoichet, Brian K.;Shoichet, Molly S.
• 通讯作者: Shoichet, Molly S.