High throughput screening of the Prp8 intein splicing inhibitors for pathogenic fungi

病原真菌 Prp8 内含肽剪接抑制剂的高通量筛选

基本信息

批准号：
10382470
负责人：
HONGMIN LI
金额：
$ 56.93万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10382470
关键词：
Affect Affinity Animal Model Antifungal Agents Antineoplastic Agents Binding Biochemical Biological Assay Biology Biophysics C-terminal Cause of Death Cells Cessation of life Chemical Structure Cisplatin Combined Modality Therapy Complex Cryptococcal Meningitis Cryptococcosis Cryptococcus Cryptococcus gattii Cryptococcus neoformans Crystallization Crystallography Development Disease Disease Outbreaks Drug Kinetics Drug Synergism Drug Targeting Drug resistance Elements Essential Genes Exteins FDA approved Genes Goals Growth HIV Human Immune Immune system Immunocompetent Immunocompromised Host In Vitro Individual Infection Lead Ligation Luciferases Lung Microbe Mutagenesis N-terminal Organ Transplantation Organism Pathogenicity Pathway interactions Peptides Persons Pharmaceutical Preparations Pharmacology Pharmacotherapy Protein Splicing Proteins Public Health RNA Splicing Resistance Severe Combined Immunodeficiency Site Speed Spliceosomes Structure Structure-Activity Relationship Testing Toxic effect Trypsin Validation Variant Work analog base chemical synthesis combat cytotoxicity drug testing fungus high throughput screening improved in silico in vivo inhibitor innovation intein invention microbial mortality new therapeutic target novel novel therapeutics pathogen pathogenic fungus resistant strain screening small molecule

项目摘要

Abstract: The fungi Cryptococcus neoformans and C. gattii cause cryptococcal meningitis (CM) and pulmonary cryptococcosis, which mainly occurs in immunocompromised people, such as those with HIV, severe combined immunodeficiency, or post organ-transplant status. Recent outbreaks of infections of C. gattii in immune competent people raise significant concerns about the overall threat of cryptococcal species to public health. There are over 1 million annual cases of CM worldwide, with estimated deaths of 700,000 per year. Treatment of CM has become increasingly difficult with a mortality rate over 50%, due to the emergence of drug-resistant strains, making the development of new treatments for cryptococcal infections imperative. Cryptococcal fungi contain protein self-splicing elements, called inteins, which are internal protein elements that self-excise from their host proteins and catalyze ligation of the flanking sequences (exteins) with a natural peptide bond. Because inteins do not exist in multi-cellular organisms, such as humans, and often disrupt the functions of critical microbial genes, they are attractive drug targets. Cryptococcal species have inteins in the essential gene, Prp8, a component of the spliceosome. Intein inhibitors have many advantages over traditional anti-fungal drugs. Because the Prp8 inteins of cryptococcal species share high sequence similarity and have similar splicing mechanisms, it is highly possible that a single intein splicing inhibitor can inhibit the splicing activities of all Prp8 inteins. Thus, Prp8 intein splicing inhibitors will be “broad-spectrum” against all Prp8 intein-containing fungal pathogens. On the other hand, because neither human nor microbes normally associated with humans have inteins, intein inhibitors would be also function as “narrow-spectrum” antifungals specific only for intein-containing pathogens such as cryptococcal species. In addition, inteins as novel drug targets have an inhibition mechanism different from those of all known drugs. Thus, intein inhibitors will provide a potentially fresh approach and may synergize with existing drugs in treating the infected. In our preliminary results, cisplatin was found not only to inhibit the intein splicing in vitro, but also to reduce the fungal burden in vivo. The major goal of this proposal is to develop and perform high throughput screening assays to identify and characterize compounds that inhibit the Prp8 intein splicing by a combination of biophysical, biochemical, computational, pharmacological, medicinal, cellular and in vivo approaches. Hit compounds with activity better than the existing drugs will be tested for reduction of growth of cryptococcal species using various assays and resistant variants. This study will generate information on inhibitor-intein interaction at structurally significant sites and potentially lead to novel therapies for cryptococcal infections.

摘要：新型隐球菌和格特隐球菌可引起隐球菌性脑膜炎（CM）和肺结核。隐球菌病，主要发生在免疫功能低下的人群中，例如艾滋病毒感染者，严重联合免疫缺陷或器官移植后状态最近爆发的格特隐球菌感染。有能力的人士对隐球菌物种对公众健康的总体威胁表示严重关切。全球每年有超过 100 万例 CM 病例，估计每年有 70 万人死亡。由于耐药性的出现，CM的治疗变得越来越困难，死亡率超过50% 菌株，使得开发隐球菌感染的新疗法势在必行。含有蛋白质自剪接元件，称为内含肽，它们是内部蛋白质元件，可以从它们的宿主蛋白并催化侧翼序列（外显子）与天然肽键的连接。内含肽不存在于多细胞生物体中，例如人类，并且经常破坏关键的功能微生物基因，它们是有吸引力的药物靶标。隐球菌物种的必需基因 Prp8 具有内含肽，剪接体的一个组成部分，与传统的抗真菌药物相比，内含肽抑制剂具有许多优点。因为隐球菌种的Prp8内含肽具有高度的序列相似性并且具有相似的剪接从机制上看，单一内含肽剪接抑制剂很可能抑制所有 Prp8 的剪接活性因此，Prp8 内含肽剪接抑制剂将对所有含有 Prp8 内含肽的真菌具有“广谱”作用。另一方面，因为人类或通常与人类相关的微生物都没有。内含肽，内含肽抑制剂也可用作仅针对含内含肽的“窄谱”抗真菌药此外，内含肽作为新的药物靶点具有抑制机制。与所有已知药物不同，因此，内含肽抑制剂将提供一种潜在的新鲜方法并可能。在我们的初步结果中，发现顺铂不仅可以与现有药物协同治疗感染者。抑制体外内含肽剪接，同时减少体内真菌负担该提案的主要目标是。开发和执行高通量筛选测定，以识别和表征抑制化合物 Prp8 内含肽剪接通过生物物理、生物化学、计算、药理学、医学、将测试活性优于现有药物的细胞和体内方法。本研究将使用各种测定法和耐药变体来减少隐球菌物种的生长。有关抑制剂与内含肽在结构上重要位点相互作用的信息，可能会带来新的治疗方法隐球菌感染。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Antiviral Agents against Flavivirus Protease: Prospect and Future Direction.

针对黄病毒蛋白酶的抗病毒药物：前景和未来方向。

DOI：
发表时间：
2022-02-25
期刊：
影响因子：
0
作者：
Samrat, Subodh K;Xu, Jimin;Li, Zhong;Zhou, Jia;Li, Hongmin
通讯作者：
Li, Hongmin

An alternative domain-swapped structure of the Pyrococcus horikoshii PolII mini-intein.

堀越火球菌 PolII 迷你内含肽的另一种结构域交换结构。

DOI：
发表时间：
2021-06-03
期刊：
影响因子：
4.6
作者：
Williams, Jennie E;Jaramillo, Mario V;Li, Zhong;Zhao, Jing;Wang, Chunyu;Li, Hongmin;Mills, Kenneth V
通讯作者：
Mills, Kenneth V

Crystal structure and transient dimerization for the FKBP12 protein from the pathogenic fungus Candida auris.

来自致病真菌耳念珠菌的 FKBP12 蛋白的晶体结构和瞬时二聚化。