Raman spectroscopic analysis of drug beta-lactamase interacteractions

药物β-内酰胺酶相互作用的拉曼光谱分析

• 批准号: 8461167
• 负责人: PAUL R CAREY
• 金额: $ 31.33万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-01 至 2014-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8461167
• 关键词: Active Sites; Antibiotic Resistance; Antibiotics; Automobile Driving; Bacteria; Bacterial Infections; Carbapenems; Catechols; Clinic; Clinical; Complement; Complex; Crystallography; Data; Data Collection; Disease Outbreaks; Drops; Drug Design; Drug resistance; Elements; Enzyme Inhibition; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Event; Exercise; Freezing; Glass; Goals; Health; Human; Imipenem; Inhibitory Concentration 50; Kinetics; Knowledge; Lactamase; Lead; Life; Life Cycle Stages; Link; Meropenem; Microscope; Microscopy; Molecular Conformation; Pathway interactions; Penicillins; Pharmaceutical Preparations; Phase; Population; Positioning Attribute; Raman Spectrum Analysis; Reaction; Reporting; Resistance; Roentgen Rays; Role; Route; Sampling; Snow; Solutions; Source; Stable Populations; Structure; Substrate Specificity; Sulfones; Technology; Therapeutic; Time; Tube; Work; X-Ray Crystallography; absorption; aqueous; bacterial resistance; base; beta-Lactamase; beta-lactamase PSE-2; chemical reaction; cold temperature; design; inhibitor/antagonist; insight; news; next generation; novel; public health relevance; research study; time use

DESCRIPTION (provided by applicant): This proposal sets out to study the chemical reactions between compounds that are good inhibitor candidates and ?-lactamase enzymes. In the clinic, bacterial infections are becoming increasingly resistant to antibiotic-based treatments. A major source of resistance is the class of ? -lactamase enzymes produced by the bacteria. These enzymes hydrolyze, and thus inactivate, penicillin-like molecules before they can attack the bacteria. Here we set out to identify the intermediates formed by mechanism-based inhibitors that block the action of clinically problematic ? -lactamases. A focal point of the study will be the elucidation of the reactions of lead inhibitors with class A and class D ? -lactamases that function as carbapenemases (KPC-2 and OXA-24) and extended spectrum ? -lactamases (SHV-2, SHV-5 and OXA 10). A major thrust involves the use of Raman crystallography, where the reactions of the inhibitors within single crystals of the enzyme are followed in real time using a Raman microscope. This provides information on the identity and conformation of intermediates on the reaction pathway as well as kinetic data on their populations. The Raman data also provide an immediate link to X-ray crystallography with its detailed structural insights, and the two approaches are highly synergistic. In addition, the Raman analysis will be extended to solution studies, linking our knowledge of the reactions in crystals to the corresponding reaction in the aqueous phase. This is important because it is becoming clear that, for some inhibitors, the results from solution studies differs from those in crystals. Initially, our studies will use lead compounds that are 6-(pyridylmethylidene) penam sulfones possessing favorable IC50 values. Our results will identify reaction intermediates that block the active site under biologically relevant conditions and provide input into the design of better inhibitors, and thence potential therapeutic compounds.

描述（由申请人提供）：该提案旨在研究良好的候选抑制剂化合物与β-内酰胺酶之间的化学反应。在临床上，细菌感染对抗生素治疗的耐药性越来越大。阻力的主要来源是？ -细菌产生的内酰胺酶。这些酶会水解青霉素样分子，从而使其失去活性，然后再攻击细菌。在这里，我们着手鉴定由基于机制的抑制剂形成的中间体，这些抑制剂可以阻止临床问题的作用？ -内酰胺酶。该研究的一个重点是阐明先导抑制剂与 A 类和 D 类药物的反应？ -具有碳青霉烯酶功能的内酰胺酶（KPC-2 和 OXA-24）和扩展谱？ -内酰胺酶（SHV-2、SHV-5 和 OXA 10）。主要推动力涉及拉曼晶体学的使用，其中使用拉曼显微镜实时跟踪酶单晶内抑制剂的反应。这提供了有关反应途径中中间体的身份和构象的信息以及其群体的动力学数据。拉曼数据还提供了与 X 射线晶体学的直接联系及其详细的结构见解，并且这两种方法具有高度的协同作用。此外，拉曼分析将扩展到溶液研究，将我们对晶体反应的了解与水相中的相应反应联系起来。这很重要，因为越来越清楚的是，对于某些抑制剂，溶液研究的结果与晶体中的结果不同。最初，我们的研究将使用具有良好 IC50 值的 6-（吡啶亚甲基）青南砜先导化合物。我们的结果将鉴定在生物学相关条件下阻断活性位点的反应中间体，并为更好的抑制剂的设计以及潜在的治疗化合物的设计提供输入。

项目成果

Sulbactam forms only minimal amounts of irreversible acrylate-enzyme with SHV-1 beta-lactamase.

舒巴坦仅与 SHV-1 β-内酰胺酶形成极少量的不可逆丙烯酸酯酶。

• DOI: 10.1021/bi7006146
• 发表时间: 2007
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Totir,MonicaA;Helfand,MarionS;Carey,MarianneP;Sheri,Anjaneyulu;Buynak,JohnD;Bonomo,RobertA;Carey,PaulR
• 通讯作者: Carey,PaulR

Deacylation and reacylation for a series of acyl cysteine proteases, including acyl groups derived from novel chromophoric substrates.

一系列酰基半胱氨酸蛋白酶的脱酰化和再酰化，包括源自新型发色底物的酰基。

• DOI: 10.1021/bi960648h
• 发表时间: 1996
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Doran,JD;Tonge,PJ;Mort,JS;Carey,PR
• 通讯作者: Carey,PR

Measuring propargyl-linked drug populations inside bacterial cells, and their interaction with a dihydrofolate reductase target, by Raman microscopy.

通过拉曼显微镜测量细菌细胞内炔丙基连接的药物群体及其与二氢叶酸还原酶靶标的相互作用。

• DOI: 10.1021/acs.biochem.5b00202
• 发表时间: 2015
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Heidari-Torkabadi,Hossein;Che,Tao;Lombardo,MichaelN;Wright,DennisL;Anderson,AmyC;Carey,PaulR
• 通讯作者: Carey,PaulR

Kinetic crystallography by Raman microscopy.

• DOI: 10.1016/j.bbapap.2010.08.006
• 发表时间: 2011-06
• 期刊: Biochimica et biophysica acta
• 作者: Carey PR;Chen Y;Gong B;Kalp M
• 通讯作者: Kalp M

A catalytic metal ion interacts with the cleavage Site G.U wobble in the HDV ribozyme.

• DOI: 10.1021/bi8020108
• 发表时间: 2009-02-24
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Chen, Jui-Hui;Gong, Bo;Bevilacqua, Philip C.;Carey, Paul R.;Golden, Barbara L.
• 通讯作者: Golden, Barbara L.