Small-Molecule Antagonists of Ral GTPases in Cancer

癌症中 Ral GTP 酶的小分子拮抗剂

基本信息

批准号：
9896783
负责人：
Samy Meroueh
金额：
$ 58.67万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9896783
关键词：
Actins Affinity Binding Bioavailable Biochemical Biological Assay Biology Biophysics Caco-2 Cells Calorimetry Cancer cell line Cell Proliferation Cell Survival Cell physiology Cells Clinic Clinical Trials Computing Methodologies Cultured Cells Cytoskeletal Modeling Data Development Docking Drug Kinetics Endocytosis Ensure Enzyme-Linked Immunosorbent Assay Evaluation Excretory function Exhibits Exocytosis Fluorescence Polarization Free Energy GTP Binding Growth Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases Human Impairment In Vitro Investigational Drugs KRAS2 gene Lead Libraries Lung Adenocarcinoma Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of urinary bladder Mediating Metabolic Metabolism Mission Mutation NMR Spectroscopy Non-Small-Cell Lung Carcinoma Oral Parents Patients Permeability Positioning Attribute Process Property Proteins Public Health RALGDS gene RNA Interference Ras Inhibitor Ras Signaling Pathway Relaxation Risk Role Signal Transduction Solubility Solvents Structure Surface Plasmon Resonance Testing Therapeutic Therapeutic Agents Titrations Toxic effect Transgenic Mice United States National Institutes of Health Work X-Ray Crystallography Xenograft Model Xenograft procedure absorption base biophysical techniques blood-brain barrier penetration cancer cell clinical investigation combinatorial design efficacy study expectation flexibility in vivo inhibitor/antagonist lung tumorigenesis mortality mouse model nanomolar novel protein protein interaction public health relevance receptor screening small molecule small molecule inhibitor tool transcription factor tumor tumor growth tumor xenograft virtual

项目摘要

DESCRIPTION (provided by applicant): Ral GTPase-regulated signaling networks control diverse set of cellular processes such as exocytosis, endocytosis, transcription factor activation, and actin cytoskeletal reorganization. Substantial evidence, including from our own work, supports a role for Ral in Ras-driven cancers such as non-small cell lung cancer (NSCLC). We found that a small molecule (BQU57) that inactivates Ral by promoting its GDP-bound state significantly impaired NSCLC proliferation in anchorage-dependent and independent assays. BQU57 was selective for Ral relative to the GTPases Ras and RhoA and inhibited tumor xenograft growth to a similar extent to the depletion of Ral using RNA interference. In addition to BQU57, we identified a small-molecule orthosteric antagonist of the protein-protein interaction between Ral and its effector protein RalBP1. The compound impairs Ral activation in NSCLC cells and inhibits proliferation in anchorage-dependent and independent assays. Our long-term objective is to develop small-molecule Ral antagonists that are suitable for treating NSCLC in the clinic. Our short-term objective is to develop derivatives of our two parental structures to probe Ral function in vivo and set the stage for clinical investigation of promising compounds. Our central hypothesis is that structure-guided computational design of derivatives of Ral antagonists will lead to small molecules that exhibit higher affinity, better pharmacokinetic (PK) properties and greater efficacy in vivo. Our preliminary data strongly positions us to test our hypothesis. BQU57 already exhibits efficacy in vivo and our orthosteric inhibitors show promising inhibition of Ral activity and cell viability in NSCLC. In our first aim, we employ structure-based computational methods and absorption, distribution, metabolism, and excretion (ADME) predictions to design derivatives for each class of small molecules. In the second aim, we synthesize 20-25 derivatives every year for each of the two structural classes and evaluate these compounds for binding and inhibition using biochemical and biophysical approaches. 1H-15N transverse relaxation-optimized (TROSY) NMR spectroscopy and X-ray crystallography are used to provide structural evidence of direct binding. In the third aim, we evaluate compounds for their effect on Ral activation, and assess their activity in a panel of NSCLC cell lines using anchorage-dependent and independent assays. We select the most promising compound for each of the two classes of Ral antagonists and characterize stability in vitro and PK parameters in vivo. Efficacy studies will be carried out first using human xenografts, followed by evaluation in transgenic mouse models for the most promising compounds. We also perform combination studies of our Ral antagonists with inhibitors of Ras signaling pathways. We expect that this work will lead to small-molecule Ral antagonists with nanomolar range affinity that are orally bioavailable, possess suitable PK properties, and exhibit substantial efficacy in blocking Ral driven tumor formation and growth.

描述（由适用提供）：RAL GTPase调节的信号网络控制各种细胞过程，例如胞吐作用，内吞作用，转录因子激活，和肌动蛋白细胞骨架重组。包括我们自己的工作在内的大量证据支持RAL在RAS驱动的癌症（例如非小细胞肺癌（NSCLC））中的作用。我们发现，通过促进其GDP结合的状态使RAL失活的小分子（BQU57）在锚固依赖性和独立测定中显着损害了NSCLC增殖。 BQU57相对于GTPases Ras和RhoA具有选择性，并在使用RNA干扰的情况下抑制了肿瘤异种移植的生长。除BQU57外，我们还确定了RAL及其效应蛋白RALBP1之间蛋白质 - 蛋白质相互作用的小分子正常拮抗剂。该化合物会损害NSCLC细胞中的RAL激活，并抑制锚定依赖性和独立测定中的增殖。我们的长期目标是开发适合在诊所治疗NSCLC的小分子RAL拮抗剂。我们的短期目标是开发两个父母结构的衍生物，以在体内探测功能，并为有希望化合物的临床研究奠定了基础。我们的中心假设是，RAL拮抗剂的衍生物的结构引导的计算设计将导致小分子暴露了更高的亲和力，更好的药代动力学（PK）特性和更高的体内效率。我们的初步数据强烈定位我们以检验假设。 BQU57已经在体内表现出效率，我们的正常抑制剂显示出有望抑制NSCLC中RAL活性和细胞生存能力的抑制作用。在我们的第一个目标中，我们采用基于结构的计算方法和分析，分布，代谢和极端（ADME）对每类小分子设计衍生物的预测。在第二个目标中，我们每年为两个结构类别中的每一个合成20-25个衍生物，并使用生化和生物物理方法评估这些化合物以结合和抑制。 1H-15N横向弛豫优化（TROSY）NMR光谱和X射线晶体学用于提供直接结合的结构证据。在第三个目标中，我们评估化合物对RAL激活的影响，并使用锚固依赖性和独立测定法评估其在NSCLC细胞系中的活性。我们为两类RAL拮抗剂选择了最有希望的化合物，并在体内表征了体外和PK参数的稳定性。功效研究将首先使用人异种移植，然后在转基因小鼠模型中评估最有前途的化合物。我们还对RAS信号通路的抑制剂进行了RAL拮抗剂的组合研究。我们预计这项工作将导致具有纳摩尔范围亲和力的小分子RAL拮抗剂，这些拮抗剂具有口服生物可利用，具有合适的PK特性，并在阻断RAL驱动的肿瘤形成和生长方面暴露了很高的效率。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A Computational Investigation of Small-Molecule Engagement of Hot Spots at Protein-Protein Interaction Interfaces.

DOI：
10.1021/acs.jcim.7b00181
发表时间：
2017-09-25
期刊：
Journal of chemical information and modeling
影响因子：
5.6
作者：
Xu D;Si Y;Meroueh SO
通讯作者：
Meroueh SO

Mimicking Intermolecular Interactions of Tight Protein-Protein Complexes for Small-Molecule Antagonists.

模拟小分子拮抗剂的紧密蛋白质-蛋白质复合物的分子间相互作用。

DOI：
10.1002/cmdc.201700572
发表时间：
2017-11-08
期刊：
ChemMedChem
影响因子：
3.4
作者：
Xu D;Bum-Erdene K;Si Y;Zhou D;Ghozayel MK;Meroueh SO
通讯作者：
Meroueh SO

Covalent Fragment Screening Identifies Rgl2 RalGEF Cysteine for Targeted Covalent Inhibition of Ral GTPase Activation.

DOI：
10.1002/cmdc.202100750
发表时间：
2022-03-18
期刊：
CHEMMEDCHEM
影响因子：
3.4
作者：
Bum-Erdene, Khuchtumur;Ghozayel, Mona K.;Xu, David;Meroueh, Samy O.
通讯作者：
Meroueh, Samy O.

Chemical Space Overlap with Critical Protein-Protein Interface Residues in Commercial and Specialized Small-Molecule Libraries.

商业和专业小分子文库中的化学空间与关键蛋白质-蛋白质界面残基重叠。

DOI：
10.1002/cmdc.201800537
发表时间：
2019
期刊：
ChemMedChem
影响因子：
3.4
作者：
Si,Yubing;Xu,David;Bum-Erdene,Khuchtumur;Ghozayel,MonaK;Yang,Baocheng;Clemons,PaulA;Meroueh,SamyO
通讯作者：
Meroueh,SamyO