Targeting Glucose Transporters Using Rapafucins

使用雷帕夫星靶向葡萄糖转运蛋白

• 批准号: 10557907
• 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 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10557907
• 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; 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; nuclear factors of activated T-cells; 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; tumor growth; uptake; 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; 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; nuclear factors of activated T-cells; 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; 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.

药物发现的主要挑战是确定小分子抑制剂 具有挑战性的靶标，包括蛋白质 - 蛋白质相互作用和多通蛋白 跨膜蛋白。大自然已经进化了解决这个问题的巧妙解决方案 如大环天然产物的免疫蛋白配体家族所示， 包括雷帕霉素和FK506。除了自己的较大尺寸以外 这些天然产品与蛋白质的相互作用更广泛，还招募了 伴侣的FKBP家庭以形成更大的二聚体复合物 与各自的靶标MTOR和钙调神经酶相关。受到启发 独特的作用方式，我们生成了一个45,000个类似雷帕霉素的库库 大环，名为Rapafucins，通过融合雷帕霉素的FKBP结合域 与组合四肽库。该库的筛选导致了 发现溶质载体（SLC）转运蛋白超级家族的多个成员， 包括葡萄糖转运蛋白（GLUT）家族的成员。两个不同的抑制剂 鉴定出Glut的rapaglutin A（RGA）和Rapaglutin E（RGE）。而RGE RGA对于GLUT1高度特异性，似乎抑制了多个GLUT同工型。 RGA 在乳腺癌的异种移植模型中，已显示出有效的 发现通过阻止DNA-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 苯甲酸苯甲酸苯甲酸苯甲酸苯甲酸苯甲酸疫苗在抑制三重阴性乳腺癌细胞方面具有协同作用 （TNBC）。我们将进一步描述协同的机制和 RGA-phenfomin组合可抑制体内TNBC肿瘤的生长。那个RGE NFAT抑制钙刺激的DNA结合活性表明潜在的作用 GLUT1调节细胞内T细胞受体信号传导。我们将阐明 RGE免疫抑制活性的基础机制并研究了 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，我们将筛选Rapafucin库与每个库 GLUT的14种同工型寻找不同同工型的特定抑制剂 Glut，它可以成为研究大量生物学的新化学工具。