Targeting Glucose Transporters Using Rapafucins

使用雷帕夫星靶向葡萄糖转运蛋白

• 批准号: 10335197
• 负责人: Jun O. Liu
• 金额: $ 33.57万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10335197
• 关键词: 3-Dimensional; Action Potentials; Affect; Amino Acids; Animals; Antineoplastic Agents; Binding; Biological Assay; Biological Availability; Biological Models; Biology; Biotin; Breast Cancer Cell; Breast Cancer Model; Calcineurin; Calcium; Cancer cell line; Cells; Chemicals; Complex; DNA Binding; Energy-Generating Resources; Exhibits; FK506; FRAP1 gene; Family; Fructose; Future; Glucose; Glucose Transporter; Growth; Immunophilins; Immunosuppression; Immunosuppressive Agents; In Vitro; Integral Membrane Protein; Lead; Libraries; Life; Ligands; Macrocyclic Compounds; Mass Spectrum Analysis; Mediating; Molecular; Molecular Chaperones; NF-kappa B; Names; Natural Products; Nature; Nutrient; Pathologic Processes; Pharmaceutical Preparations; Phenformin; Physiological Processes; Play; Post-Translational Protein Processing; Protein Dephosphorylation; Protein Isoforms; Proteins; Proteomics; Receptor Signaling; Reporter Genes; Resistance; Role; SLC2A1 gene; Series; Signal Transduction; Sirolimus; Site; Skin Transplantation; Specificity; Sum; T Cell Receptor Signaling Pathway; T-Cell Receptor; T-Lymphocyte; Tacrolimus Binding Proteins; Testing; Transcription Factor AP-1; Xenograft Model; Xenograft procedure; aerobic glycolysis; analog; anti-cancer; cancer cell; combinatorial; dimer; drug discovery; glucose uptake; in vivo; inhibitor; insight; mTOR Signaling Pathway; malignant breast neoplasm; member; metabolome; metabolomics; mutant; novel; overexpression; protein protein interaction; recruit; screening; side effect; small molecule inhibitor; solute; synergism; tool; transcription factor; translational potential; transplant model; triple-negative invasive breast carcinoma; tumor growth; uptake

A major challenge in drug discovery is to identify small molecule inhibitors for the challenging targets including protein-protein interactions and multi-pass transmembrane proteins. Nature has evolved an ingenious solution to this problem as exemplified by the immunophilin ligand family of macrocyclic natural products, including rapamycin and FK506. Aside from their own larger sizes that enable more extensive interactions with proteins, these natural products also recruit the FKBP-family of chaperones to form much larger dimeric complexes before associating with their respective targets, mTOR and calcineurin. Inspired by this unique mode of action, we generated a 45,000-compound library of rapamycin-like macrocycles, named rapafucins, by fusing the FKBP-binding domain of rapamycin with a combinatorial tetrapeptide library. Screening of this library has led to the discovery of multiple members of the solute carrier (SLC) transporter superfamily, including members of the glucose transporter (GLUT) family. Two distinct inhibitors of GLUT were identified, rapaglutin A (RgA) and rapaglutin E (RgE). Whereas RgE is highly specific for GLUT1, RgA seems to inhibit multiple GLUT isoforms. RgA has been shown to be efficacious in a xenograft model of breast cancer while RgE was found to inhibit intracellular T cell receptor signaling by blocking the DNA- binding activity of NFAT. We will further characterize the antitumor activity of RgA by systematically assessing its effect on cellular metabolomic profiles and the AMPK-mTOR signaling pathway. In preliminary studies, we have found that RgA and phenformin have synergy in inhibiting triple negative breast cancer cells (TNBC). We will further delineate the mechanism of synergy and the effect of the RgA-phenfomin combination for inhibiting TNBC tumor growth in vivo. That RgE inhibited calcium stimulated DNA-binding activity of NFAT suggests a potential role of GLUT1 in regulating intracellular T cell receptor signaling. We will elucidate the mechanism underlying the immunsuppressive activity of RgE and investigate the potential of RgE as an immunosuppressant in vivo. Lastly, using a newly developed microarray platform, we will screen rapafucin libraries against each of the 14 isoforms of GLUTs in search of specific inhibitors of different isoforms of GLUT, which can become new chemical tools for studying the biology of GLUT.

药物发现的一个主要挑战是识别小分子抑制剂 具有挑战性的目标，包括蛋白质-蛋白质相互作用和多通道 跨膜蛋白。大自然已经进化出一个巧妙的解决方案来解决这个问题 如大环天然产物的亲免素配体家族所示， 包括雷帕霉素和FK506。除了它们自身较大的尺寸之外， 与蛋白质更广泛的相互作用，这些天然产物还招募 FKBP-分子伴侣家族在形成更大的二聚体复合物之前 与各自的目标 mTOR 和钙调神经磷酸酶相关联。受此启发 独特的作用模式，我们生成了 45,000 种雷帕霉素类化合物库 通过融合雷帕霉素的 FKBP 结合结构域，命名为 rapafucins 大环化合物 具有组合四肽文库。该文库的筛选导致 发现溶质载体（SLC）转运蛋白超家族的多个成员， 包括葡萄糖转运蛋白 (GLUT) 家族的成员。两种不同的抑制剂 确定了 GLUT 的雷帕凝集素 A (RgA) 和雷帕集素 E (RgE)。而RgE RgA 对 GLUT1 具有高度特异性，似乎可以抑制多种 GLUT 同工型。红细胞A 已被证明在乳腺癌异种移植模型中有效，而 RgE 发现通过阻断 DNA- 来抑制细胞内 T 细胞受体信号传导 NFAT 的结合活性。我们将进一步表征RgA的抗肿瘤活性 通过系统地评估其对细胞代谢组谱的影响和 AMPK-mTOR 信号通路。在初步研究中，我们发现RgA 与苯乙双胍协同抑制三阴性乳腺癌细胞 （TNBC）。我们将进一步阐明协同作用的机制和效果 RgA-phenfomin 组合用于抑制体内 TNBC 肿瘤生长。那个RgE 抑制钙刺激的 NFAT DNA 结合活性表明其潜在作用 GLUT1 在调节细胞内 T 细胞受体信号传导中的作用。我们将阐明 RgE 免疫抑制活性的机制并研究 RgE 作为体内免疫抑制剂的潜力。最后，使用新的 开发的微阵列平台，我们将针对每个筛选雷帕夫星文库 GLUT 的 14 种同工型，寻找不同同工型的特异性抑制剂 GLUT，可以成为研究GLUT生物学的新化学工具。