Advancing Small Molecule Read Through Compounds to Prevent and/or Treat Heritable Pulmonary Artery Hypertension

推进小分子化合物读取以预防和/或治疗遗传性肺动脉高血压

基本信息

批准号：
10011012
负责人：
VIRENDER K REHAN
金额：
$ 25.21万
依托单位：
ADVENT THERAPEUTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-15 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10011012
关键词：
Activities of Daily Living Acute Address Aftercare Amino Acids Anatomy Antibiotics BMPR2 gene Biochemical Biological Assay Blood Blood Cells Bone Morphogenetic Protein Receptor Gene Cardiovascular Diseases Catheterization Cell membrane Cell surface Cells Cellular Assay Cessation of life Characteristics Childhood Chronic Chronic Toxicity Tests Collaborations DNA Sequence Alteration Data Development Disease Dose Endothelial Cells Endothelium Exhibits Functional disorder Gene Targeting Genes Heart Heart failure Hereditary Disease Heritability Human In Vitro Individual Lead Legal patent Length Life Lung Medical Messenger RNA Methods Mus Mutation Names Neonatal Nonsense Mutation Orphan Drugs Pathology Pathway interactions Patients Permeability Pharmaceutical Preparations Phosphorylation Positioning Attribute Prevalence Production Proteins Publishing Pulmonary Hypertension Pulmonary artery structure Pulmonary vessels Rare Diseases Research Personnel Ribosomes Rodent Series Signal Pathway Signal Transduction Site Smooth Muscle Myocytes Solubility Terminator Codon Testing Therapeutic Tissues Toxic effect Transcript Treatment Effectiveness Treatment Efficacy Universities Up-Regulation base bone morphogenetic protein receptors cadherin 5 clinically relevant dalton effective therapy efficacy testing experimental study hemodynamics high throughput screening improved in vitro Assay in vivo mRNA Expression metabolic profile mortality mouse model multidisciplinary mutant novel therapeutics patient population premature pressure prevent protein expression pulmonary arterial hypertension receptor receptor expression reflectance confocal microscopy small molecule small molecule libraries

项目摘要

Advent Therapeutics is a company that develops new therapies for neonatal, pediatric, and Orphan Drug “niche” markets to address serious unmet medical needs. Our team proposes early stage development of a new treatment for hereditable pulmonary arterial hypertension (hPAH), a rare, hereditary disorder that involves progressive elevation of pulmonary arterial pressures, right heart failure, and death. There are no effective treatments. hPAH is often caused by premature termination codons (PTCs) in the mRNA encoded by nonsense mutations in the type II bone morphogenetic protein receptor (BMPR2) gene. Certain antibiotics possess the capacity to induce “readthrough” of PTCs and thus produce a full-length protein, but are unacceptably toxic. To obviate this toxicity, we first performed a high-throughput screen on a 70,000-compound small molecule library, using a readthrough assay that indicated hits. We derivatized and patented hits, achieved in vitro, ex vivo, and in vivo proof-of-concept using PTCs in disease-causing genes other than BMPR2, and published these results. We next selected a molecule named GJ103 from among the derivatized compounds for further development, based on readthrough efficiency and favorable physical and solubility characteristics, and began testing in hPAH, using blood outgrowth endothelial cells obtained from patients with hPAH that contained disease-relevant PTCs in BMPR2. GJ103 treatment induced significant expression of BMPR2 protein. Further testing with mouse cells corroborated the results with human cells. Next, we collaborated with the Morell lab (University of Cambridge, UK) to test GJ103 in mice they created that develop hPAH by virtue of one of two clinically-relevant nonsense mutations in Bmpr2 (Bmpr2+/R899X and Bmpr2+/R584X mice). After performing preliminary dose-finding studies, GJ103 induced up to half of wild-type BMPR2 protein levels in vivo. Preliminary acute and chronic toxicity testing failed to show evidence of toxicity. BMPR2 protein induced by GJ103 demonstrated its functional effects in an LPS permeability assay in vitro and in vivo, and confocal microscopy studies indicated the protein appeared to traffic properly to the cell surface of endothelial and smooth muscle cells. Downstream upregulation of BMP signaling intermediaries (SMADs and phosphoSMADs) and BMP pathway gene targets (Id1, Id2, and VE- cadherin) indicated functional activation of BMP signaling. Here we propose to test the critical question for using GJ103 to treat hPAH: can GJ103-induced expression of BMPR2 prevent or slow development of hPAH in Bmpr2+/R899X or Bmpr2+/R584X mice if treatment is initiated early enough? In Aim 1, we will collaborate with the Soban lab (UCLA) to test whether GJ103 can induce expression of BMPR2 sufficient to slow or halt development of hPAH in mice. Dependent variables include testing right heart hemodynamics and remodeling using echo and direct catheterization in Bmpr2+/R899X and Bmpr2+/R584X mice treated with GJ103. Aim 2 proposes concomitant expression analyses with the Rehan lab (LA BioMed) of BMPR2 protein, which should be elevated if GJ103 is successful. Our studies might represent the first hope for individuals suffering from hPAH.

Advent Therapeutics 是一家开发新生儿、儿科和孤儿药新疗法的公司我们的团队提出了一种新的早期开发方案，旨在解决严重的未满足的医疗需求。遗传性肺动脉高压 (hPAH) 的治疗，这是一种罕见的遗传性疾病，涉及肺动脉压进行性升高，右心衰竭，而死亡尚无特效药。 hPAH 通常是由无义编码的 mRNA 中的过早终止密码子 (PTC) 引起的。 II 型骨形态发生蛋白受体 (BMPR2) 基因的突变某些抗生素具有该突变。诱导 PTC“通读”从而产生全长蛋白质的能力，但具有不可接受的毒性。为了避免这种毒性，我们首先对 70,000 种化合物的小分子库进行了高通量筛选，使用指示命中的通读分析，我们对命中进行了衍生和专利化，在体外、离体和体外实现。在 BMPR2 以外的致病基因中使用 PTC 进行体内概念验证，并发表了这些结果。接下来我们从衍生化合物中选择了一种名为 GJ103 的分子进行进一步开发，基于通读效率和有利的物理和溶解度特性，并开始在 hPAH 中进行测试，使用从 hPAH 患者获得的血液生长内皮细胞，其中含有与疾病相关的 PTC 在 BMPR2 中，GJ103 治疗诱导了 BMPR2 蛋白的显着表达。接下来，我们与莫雷尔实验室（剑桥大学）合作证实了这一结果。英国）在他们创造的小鼠中测试 GJ103，这些小鼠由于两个临床相关的无意义之一而发展出 hPAH Bmpr2（Bmpr2+/R899X 和 Bmpr2+/R584X 小鼠）的突变在进行初步剂量探索研究后， GJ103 体内诱导的 BMPR2 蛋白水平高达野生型的一半。未能证明 GJ103 诱导的 BMPR2 蛋白具有其功能效应。体外和体内 LPS 渗透性测定以及共聚焦显微镜研究表明该蛋白质似乎 BMP 的下游上调。信号传导中介体（SMAD 和磷酸化 SMAD）和 BMP 通路基因靶标（Id1、Id2 和 VE- 钙粘蛋白）表明 BMP 信号传导的功能激活，在此我们建议测试使用的关键问题。 GJ103治疗hPAH：GJ103诱导的BMPR2表达能否预防或减缓hPAH的发展 Bmpr2+/R899X 或 Bmpr2+/R584X 小鼠是否尽早开始治疗？在目标 1 中，我们将与 Soban 实验室 (UCLA) 测试 GJ103 是否可以诱导 BMPR2 的表达，足以减缓或停止小鼠 hPAH 的发展因变量包括测试右心血流动力学和重构。在接受 GJ103 建议治疗的 Bmpr2+/R899X 和 Bmpr2+/R584X 小鼠中使用回声和直接导管插入术。与 Rehan 实验室 (LA BioMed) 进行 BMPR2 蛋白的伴随表达分析，该蛋白应该升高如果 GJ103 成功，我们的研究可能为 hPAH 患者带来第一个希望。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Maternal food restriction-induced intrauterine growth restriction in a rat model leads to sex-specific adipogenic programming.

DOI：
10.1096/fj.202000985rr
发表时间：
2020-12
期刊：
FASEB journal : official publication of the Federation of American Societies for Experimental Biology
影响因子：
0
作者：
Sreekantha S;Wang Y;Sakurai R;Liu J;Rehan VK
通讯作者：
Rehan VK

Perinatal exposure to nicotine alters spermatozoal DNA methylation near genes controlling nicotine action.