Slow release of isotopically labelled hydrogen sulfide to track its location in animals and tissues

缓慢释放同位素标记的硫化氢以追踪其在动物和组织中的位置

基本信息

批准号：
10382220
负责人：
NED B BOWDEN
金额：
$ 7.59万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-05 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10382220
关键词：
Adhesions Affect Alzheimer&apos s Disease Amino Acids Animal Disease Models Animal Model Animals Arthritis Autoradiography Biology Cell membrane Cells Chemicals Cysteine Degenerative polyarthritis Development Disease Dose Enzymes Fluorescence Spectroscopy Glaucoma Glutathione Goals Half-Life Heart failure Human Hydrogen Sulfide Hypertension Image Individual Integral Membrane Protein Investigation Isotope Labeling Label Leukocytes Location Malignant Neoplasms Mass Spectrum Analysis Medicine Methionine Methods Myocardial Infarction Neuroglia Neurons Organ Outcome Parkinson Disease Pathway interactions Persons Pharmaceutical Preparations Phase Phase I Clinical Trials Phase II Clinical Trials Proteins Public Health Radioactive Reaction Reactive Oxygen Species Reagent Research Rheumatoid Arthritis Scientist Sepsis Signal Pathway Stimulus Structure Sulfate Sulfhydryl Compounds Testing Time Tissue Model Tissues Work animal tissue base biological adaptation to stress cell type chemical release chemical synthesis clinical application clinical development diallyl trisulfide endoplasmic reticulum stress in vivo innovation light microscopy mouse model neuron loss novel therapeutics persulfides phase II trial radiotracer response success therapy development

项目摘要

Hydrogen sulfide is a key gasotransmitter in human cells, and the addition of exogenous hydrogen sulfide at single micromolar or lower concentrations is actively being explored for clinical applications. Three drugs based on the release of hydrogen sulfide are in phase I or II clinical trials. Although the location of where exogenous hydrogen sulfide partitions inside of cells can be tracked using several two-step tagging methods, tracking the location of exogenous hydrogen sulfide in tissue or animal models cannot be done. This limits the development of therapies based on the slow release of hydrogen sulfide because it cannot be tracked once it is administered to an animal or tissue. The long-term goal is to investigate the mechanisms of how exogenous hydrogen sulfide is transported and accumulates in the body. The overall objective of this application is to synthesize chemicals widely used to add exogenous hydrogen sulfide, so these chemicals release radioactive 35S labelled hydrogen sulfide which will allow it to be tracked on a tissue and animal level for the first time. The central hypothesis is that 35S labeled sulfate can be converted into several key chemicals that release hydrogen sulfide in response to different stimuli, and that the concentration and amount of 35S labelled hydrogen sulfide can be rationally controlled. This proposal builds on prior work by others that used 35S labelled methionine, cysteine, and other chemicals to investigate where they partitioned in animal and tissue models using scintillation and light microscopy autoradiography. Despite the success of the use of these 35S labelled chemicals, no one has described the synthesis of chemicals that slowly release 35S labelled hydrogen sulfide despite its importance in medicine. The rationale for completing this work is that it will allow scientists to track where exogenous hydrogen sulfide partitions in tissue and animal models for the first time to thoroughly investigate the origins of the effect of exogenous hydrogen sulfide in treating diseases in animal models. The central hypothesis will be tested by pursuing two specific aims: 1) Identify the reactions to synthesize GYY- 4137, dialkyldithiophosphates, caged perthiols, and DTT to release 35S labelled hydrogen sulfide at controlled rates and doses; and 2) Identify the reactions to synthesize diallyl trisulfide and trisulfides of cysteine and glutathione that will release 35S labelled hydrogen sulfide in the presence of thiols. The research proposed in this application is innovative, in the applicant's opinion, because it challenges the status quo of tracking the location of exogenous hydrogen sulfide by the synthesis of chemicals that will allow it to be tracked on the tissue and animal levels for the first time. This proposal combines scintillation and light microscopy autoradiography that were used to image 35S labelled amino acids with the first synthesis of chemicals to slowly release 35S labelled hydrogen sulfide to solve a key problem in biology. The proposed research is significant because it will advance research already underway to understand the effects of exogenous H2S and to more rapidly develop clinical applications of these chemicals.

硫化氢是人类细胞中的关键焦虑剂，在为临床应用积极探索单微摩尔或较低浓度。三种药物根据硫化氢的释放，在I期或II期临床试验中。虽然位置的位置可以使用几种两步标记方法跟踪细胞内部的外源性硫化氢分区，无法完成跟踪硫化物在组织或动物模型中的外源性氢的位置。这限制了基于硫化氢的缓慢释放的疗法开发，因为它一旦无法跟踪用于动物或组织。长期目标是研究外源的机制硫化氢被运输并积聚在体内。该应用程序的总体目的是合成化学品广泛用于添加外源性硫化物的化学物质，因此这些化学物质释放了放射性 35S标记为硫化氢，将首次在组织和动物水平上跟踪它。这中心假设是35s标有硫酸盐可以转换为几种释放的关键化学物质硫化氢响应于不同的刺激，并标记为35s的浓度和量硫化氢可以合理控制。该提案建立在使用35s的其他人的先前工作的基础上标记为蛋氨酸，半胱氨酸和其他化学物质，以研究它们在动物和组织中分配的位置使用闪烁和光显微镜放射自显影的模型。尽管使用了这35秒标有化学物质，没有人描述化学物质的合成，该化学物质慢慢释放了35s的氢硫化物尽管在医学上很重要。完成这项工作的理由是，它将允许科学家跟踪第一次在组织和动物模型中的外源性硫化物分区以彻底彻底研究硫化物外源性氢在动物模型中治疗疾病中的作用的起源。这将通过追求两个具体目的来检验中心假设：1）确定对gyy-合成的反应 4137，dialkyldithiophathates，笼中的perthiols和DTT，以在受控的价格和剂量； 2）确定对半胱氨酸和半胱氨酸的三硫化物和三硫化物的反应谷胱甘肽将在存在硫醇的情况下释放35s标记的硫化氢。该研究提出了申请人认为，该应用程序具有创新性，因为它挑战了跟踪的现状通过合成化学物质的硫化物外源性氢的位置，将其跟踪第一次组织和动物水平。该建议结合了闪烁和光学显微镜放射自显影用于图像35s标记的氨基酸，并首先合成化学物质慢慢释放35S标记硫化氢，以解决生物学中的关键问题。拟议的研究是意义重大，因为它将推进已经正在进行的研究，以了解外源H2S的影响和更快地开发了这些化学物质的临床应用。