黄嘌呤氧化酶抑制剂的研究进展

痛风症是一种因嘌呤代谢紊乱导致尿酸生成异常的常见病。以黄嘌呤氧化酶为靶点的药物是一类重要的抗痛风药物。有效抑制黄嘌呤氧化酶的活性可减少黄嘌呤向尿酸的转化,从而降低体内尿酸的水平,达到治疗痛风的目的。本文就近几年黄嘌呤氧化酶抑制剂的研究进展进行综述。     

高尿酸血症治疗药物黄嘌呤氧化酶抑制剂的研究进展

高尿酸血症是由多种原因引起的代谢性疾病,黄嘌呤氧化酶(XO)是药物治疗高尿酸血症的重要靶点。随着黄嘌呤氧化酶晶体结构的解析、计算机辅助药物设计和高通量筛选等技术的运用,近年涌现了许多具有良好的XO抑制活性的化合物。中药也是我国治疗高尿酸血症的一种主要方式,对天然化合物进行结构修饰也是寻找新化合物的重要方法。该文基于高尿酸血症的产生机制,对具有良好XO抑制活性化合物进行综述。     

黄嘌呤氧化酶抑制剂/超氧阴离子清除剂双靶点高通量筛选模型的建立

本文建立了适用于同时筛选黄嘌呤氧化酶抑制剂和超氧阴离子清除剂的双靶点高通量复合筛选模型。在黄嘌呤氧化酶超氧阴离子生成体系中,加入WST-1作为超氧阴离子生成量的探针,以反应体系中标识性产物尿酸为黄嘌呤氧化酶活性指示剂,采用Spectra Max M5酶标测试仪,同时检测两种指示剂的浓度变化,通过对反应体系中的影响因素进行优化建立双靶点HTS筛选模型,并利用阳性药物对该模型进行评价。在反应体系中,反应终体积50μL,黄嘌呤氧化酶4 m U·m L-1、黄嘌呤250μmol·L-1、WST-1浓度为100μmol·L-1,黄嘌呤氧化酶抑制剂筛选模型的Z’-因子为0.537 4,S/N为47.519 9;超氧阴离子清除剂筛选模型的Z’-因子为0.507 4,S/N为5.388 9。结果表明,本文建立的黄嘌呤氧化酶抑制剂/氧自由基清除剂高通量筛选模型具有稳定性好、成本较低和重复性高等特点,可以广泛应用于药物筛选。     

黄嘌呤氧化酶抑制剂的研究进展

黄嘌呤氧化还原酶（Xanthine oxidoreductase,XOD)是一种高度通用的黄素酶,在不同物种之间以及生物各种组织中普遍存在。痛风是由于体内尿酸代谢障碍,导致尿酸水平过高,影响机体的正常运行。抑制黄嘌呤氧化还原酶可以降低尿酸水平,发挥抗高尿酸的作用。现如今临床上用于治疗痛风、高尿酸血症最有效的药物还是黄嘌呤氧化还原酶抑制剂。本文归纳了黄嘌呤氧化还原酶抑制剂的药理作用和临床应用,并对近年来已经上市和处于开发阶段的黄嘌呤氧化还原酶抑制剂进行总结,综述了黄嘌呤氧化还原酶抑制剂的最新进展。     

菊花中的黄嘌呤氧化酶抑制剂

黄嘌呤氧化酶（XO）是将黄嘌呤和次黄嘌呤氧化催化为尿酸引起高尿酸血症和痛风病的关键酶。为寻找新的植物来源的XO抑制剂，作者在筛选288种植物提取物中，     

金线莲提取物对黄嘌呤氧化酶活性及高尿酸血症小鼠的影响

目的 研究金线莲提取物体外抑制黄嘌呤氧化酶活性的作用和体内降尿酸的作用.方法 用黄嘌呤氧化酶(XOD)试剂盒测定不同浓度金线莲乙醇提取物和水提取物对黄嘌呤氧化酶活性的抑制作用;采用小鼠高尿酸血症模型研究了金线莲水提物对尿酸和肌酐水平的影响.结果 金线莲水提物对黄嘌呤氧化酶(XOD)具有较强的抑制作用,金线莲水提物低、高剂量组均显著降低血清尿酸水平(P<0.01)和血清肌酐水平(WTBXP<0.01).结论 金线莲水提物对小鼠高尿酸血症模型具有一定的治疗作用.     

分子对接结合体外筛选挖掘来自中药的黄嘌呤氧化酶抑制剂

目的:基于分子对接和体外活性实验策略挖掘抑制黄嘌呤氧化酶(xanthine oxidase, XO)的中药活性分子,以期获得活性较好的先导化合物。方法:以Auto dock分子对接软件对传统中药数据库中6味常用清热利湿药的137个化合物与XO进行分子模拟对接,然后运用体外酶抑制活性实验对分子模拟对接结果较好的化合物进行二次筛选。结果:本研究首先以XO为靶点对中医治疗痛风过程中的6味常用清热利湿药黄柏、知母、忍冬藤、赤芍、虎杖和葎草中共137个化合物进行对接模拟,从中挑选13个打分值较高的化合物进行体外酶活性筛选,最终明确了槲皮万寿菊苷、去氧胆酸和石蒜碱3个化合物对XO蛋白酶具有较好的抑制活性。其中槲皮万寿菊苷、去氧胆酸和石蒜碱的半数抑制浓度(IC₅₀)值分别是19.18,24.47,27.78μmol·L^-1。结论:从6味常用清热利湿中药中挖掘到槲皮万寿菊苷、去氧胆酸和石蒜碱3个XO抑制剂潜在活性分子,可以作为XO抑制剂发挥抗痛风作用。     

黄嘌呤氧化酶抑制剂非布司他衍生物的研究进展

黄嘌呤氧化酶抑制剂非布司他是经典的抗痛风药物,治疗效果显著且耐受性好.非布司他及其衍生物在结构上可以分为两个主要部分,分别为取代苯环和羧基取代的五元或六元杂环.本文综述了近10年来非布司他衍生物的研究进展,针对各类衍生物的构效关系进行了分类概述.探究黄嘌呤氧化酶抑制剂的作用机制和构效关系,对合理设计和开发抗痛风的新化学...     

茵连痛风颗粒中黄嘌呤氧化酶抑制剂的虚拟筛选

目的 探究中药复方茵连痛风颗粒中对黄嘌呤氧化酶有抑制作用的活性成分。方法 依据文献报道,建立了茵连痛风颗粒中所含化合物的结构数据库;以黄嘌呤氧化酶为靶标,使用AutoDock Vina 软件对这些化合物进行虚拟筛选,并用AutoDock Tool对代表性活性成分与黄嘌呤氧化酶的作用模式进行了分析。结果 茵连痛风颗粒所含物质中有13个化合物与黄嘌呤氧化酶的结合能在-9.0 kcal·mol^-1以下,其中主要为黄酮类成分。结合模式分析表明这些活性分子可以与黄嘌呤氧化酶的活性位点形成-相互作用、疏水相互作用和氢键相互作用。结论 茵连痛风颗粒中含有黄嘌呤氧化酶抑制剂,可通过降低尿酸的生成发挥抗痛风作用。     

黄嘌呤氧化还原酶抑制剂的构效关系研究进展

介绍了近年来黄嘌呤氧化还原酶及其抑制剂的研究进展。从构效关系的角度出发,概述与黄嘌呤氧化还原酶底物结构类似的抑制剂和非嘌呤类抑制剂的特点,为新的黄嘌呤氧化还原酶抑制剂的研发提供参考。     

Establishment of a high-throughput screening system for caspase-3 inhibitors

In most tissues, apoptosis plays a pivotal role in normal development and for regulating cell number, thus inappropriate apoptosis underlies a variety of diseases. Caspase-3 is one of a family of caspases that are mainly involved in the apoptotic signal transduction pathway, where caspase-3 acts as an effect molecule to proteolytically cleave intracellular substrates that are necessary for maintaining cell survival. Recent evidences show that apoptotic cell death can be blocked by inhibiting caspase-3, suggesting its inhibitors have potential to be therapeutic drugs for the diseases related with inappropriate apoptosis. We have established a screening system to search caspase-3 inhibitors from chemical libraries stocked in our institute. The enzyme assay is configured entirely in 96-well format, which is easily adapted for high throughput screening. Before performing mass screening, 80 in-house compounds were screened as a preliminary experiment, and we found that morin hydrate inhibited caspase-3 by 66.4% at the final concentration of 20 microM.     

Progress towards the discovery of xanthine oxidase inhibitors

Xanthine oxidase (XO) is a highly versatile flavoprotein enzyme, ubiquitous among species (from bacteria to human) and within the various tissues of mammals. The enzyme catalyses the oxidative hydroxylation of purine substrates at the molybdenum centre (the reductive half-reaction) and subsequent reduction of O(2) at the flavin centre with generation of reactive oxygen species (ROS), either superoxide anion radical or hydrogen peroxide (the oxidative half-reaction). Many diseases, or at least symptoms of diseases, arise from a deficiency or excess of a specific metabolite in the body. For an example of an excess of a particular metabolite that produces a disease state is the excess of uric acid which can led to gout. Inhibition of XO decreases the uric acid levels, and results in an antihyperuricemic effect. Allopurinol, first synthesised as a potential anticancer agent, is nowadays a clinically useful xanthine oxidase inhibitor used in the treatment of gout. There is overwhelming acceptance that xanthine oxidase serum levels are significantly increased in various pathological states like hepatitis, inflammation, ischemia-reperfusion, carcinogenesis and aging and that ROS generated in the enzymatic process are involved in oxidative damage. Thus, it may be possible that the inhibition of this enzymatic pathway would be beneficial. In this review the State of the Art will be presented, which includes a summary of the progress made over the past years in the knowledge of the structure and mechanism of the enzyme, associated pathological states, and in the efforts made towards the development of new xanthine oxidase inhibitors.     

BACE1抑制剂分子水平高通量筛选体系的建立

目的建立体外分子水平β-分泌酶（BACE1）抑制剂高通量筛选体系。方法采用均相时间分辨荧光法（HTRF）,通过优化反应时间、酶浓度、检测仪器等实验条件,建立BACE1抑制剂高通量筛选体系,根据待测样品对BACE1活性抑制程度筛选其抑制剂。结果采用HTRF法建立了BACE1抑制剂高通量筛选体系,其中反应时间确定为6h、BACE1浓度为670U/L,采用VICTOR3酶标仪测得信噪比为649.6,信背比为44.6,Z′因子为0.91,变异系数小于10%,并筛选到一系列IC50小于10-6mol/L的化合物。结论采用HTRF法建立的BACE1抑制剂筛选体系灵敏度高、特异性和稳定性好,可用于BACE1抑制剂的高通量筛选。     

Therapeutic voyage of synthetic and natural xanthine oxidase inhibitors

Xanthine oxidase (XO) inhibitors are commonly used to treat gout, nephropathy, and renal stone diseases related to hyperuricemia. However, recent research has shown that these inhibitors may also have potential benefits in preventing vascular diseases, including those affecting the cerebrovasculature. This is due to emerging evidence suggesting that serum uric acid is involved in the growth of cardiovascular disease, and XO inhibition can reduce oxidative stress in the vasculature. There is a great interest in the development of new XO inhibitors for the treatment of hyperuricemia and gout. The present review discusses the many synthetic and natural XO inhibitors that have been developed which are found to have greater potency.     

2-Alkyloxyalkylthiohypoxanthines as new potent inhibitors of xanthine oxidase.

The title compounds were prepared and tested as xanthine oxidase (XO) inhibitors. Results evidenced that potency was related to the position of the oxygen atom in the 2-linear chain and that it grew with distance from the sulfur atom until it became equipotent to 2-n-hexylthiohypoxanthine. Enzymatic oxidation on C(2) occurred in the 8-alkylthiohypoxanthines. On the contrary, oxidation on C(8) did not occur in the 2-alkythioderivatives, demonstrating that the chain forced these molecules to form a complex with molybdenum(VI) involving only the N(3) and N(9) nitrogen atoms.     

An updated patent review: xanthine oxidase inhibitors for the treatment of hyperuricemia and gout (2011-2015)

Introduction: Xanthine oxidase (XO) is a versatile molybdoflavoprotein, widely distributed, occurring in milk, kidney, lung, heart, and vascular endothelium. Catalysis by XO to produce uric acid and reactive oxygen species leads to many diseases. Anti hyperuricemic therapy by xanthine oxidase inhibitors has been mainly employed for the treatment of gout.

Area covered: This review covers the patent literature (2011–2015) and also presents the interesting strategies/rational approaches employed for the design of xanthine oxidase inhibitors reported recently.

Expert opinion: Recent literature indicates that various non purine scaffolds have been extensively investigated for xanthine oxidase inhibition. The significant potential endowed by heteroaryl based compounds, in particularly fused heterocycles clearly highlights their clinical promise and the need for detailed investigation. Studies by various research groups have also revealed that the flavone framework is open for isosteric replacements and structural modifications for yielding potent non purine xanthine oxidase inhibitors. In addition, various plant extracts recently reported to possess significant xanthine oxidase inhibitory potential presents enough promise to initiate a screening program for the identification of other plant extracts and phytoconstituents possessing inhibitory potential towards the enzyme.