Inhibitors of Phosphatidylinositol-specific Phospholipase C with Myo-inositol Scaffold.

IF 1.9 4区 医学 Q3 CHEMISTRY, MEDICINAL
Christian Bierkamp, Walburga Hanekamp, Christoph Arenz, Matthias Lehr
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引用次数: 0

Abstract

Background: Phosphatidylinositol-specific phospholipase C (PI-PLC) enzymes catalyze the conversion of phosphatidylinositol-4,5-bisphosphate into the second messengers diacylglycerol and inositol- 1,4,5-trisphosphate, both of which play crucial roles in regulating biochemical processes. Despite the wellestablished link between elevated PI-PLC activity and pathophysiological conditions, no PI-PLC inhibitors are currently in clinical development. Moreover, existing inhibitors demonstrate only limited potency.

Objective: Due to the structural similarity with known inhibitors with a myo-inositol backbone, DL-1-Ododecylsulfonyl- myo-inositol-3,5-bisphosphate, designated as acid sphingomyelinase inhibitor, and derivatives thereof should be tested for inhibition of PI-PLC activity.

Methods: The newly synthesized compounds were evaluated for their ability to inhibit PI-PLC activity in porcine platelet lysate and porcine brain homogenate, as well as their inhibitory potency against the recombinant isoenzymes PLCγ1 and PLCγ2. The assay measured the release of diacylglycerol from L-α- phosphatidylinositol using HPLC coupled with MS detection. Furthermore, the specificity of selected compounds was assessed by determining their inhibitory potency against other surface-active enzymes through HPLC-based assays.

Results: It was found that DL-1-O-dodecylsulfonyl-myo-inositol-3,5-bisphosphate inhibits PI-PLC activity at micromolar concentrations. However, its maximum achievable inhibitory effect was limited to approximately 70%. Through structural modifications, inhibitors were developed that led to near complete inhibition of PIPLC activity. The study also revealed that the alleged PI-PLC inhibitor U73122, still frequently cited in the literature to demonstrate PI-PLC involvement in biochemical processes, is unsuitable for this purpose. Consistent with observations by others, its inhibitory activity in bionucleophile-containing cell or tissue preparations was found to be significantly lower than its activity against purified PI-PLC enzymes. Additionally, U73122 was shown to inhibit other enzymes, such as cytosolic phospholipase A2α, fatty acid amide hydrolase, and monoacylglycerol lipase, which, like PI-PLC, metabolize lipophilic substrates. In contrast, the newly developed myo-inositol derivatives exhibited reduced sensitivity to bionucleophiles and significantly improved selectivity against the tested surface-active enzymes compared to U73122.

Conclusion: New compounds exhibiting significant inhibitory activity against PI-PLC have been identified. The findings could prove valuable in the development of clinically applicable PI-PLC inhibitors, particularly for the treatment of cancer. Additionally, the myo-inositol derivatives developed demonstrated greater suitability for studying PI-PLC's role in physiological processes in tissue homogenates compared to the maleimide derivative U73122, which is commonly used for this purpose in scientific research. This advantage arises from the fact that U73122 is a non-specific 'pan-assay interference compound' (PAIN).

肌醇支架对磷脂酰肌醇特异性磷脂酶C的抑制作用。
背景:磷脂酰肌醇特异性磷脂酶C (PI-PLC)酶催化磷脂酰肌醇-4,5-二磷酸转化为第二信使二酰基甘油和肌醇- 1,4,5-三磷酸,两者在调节生化过程中起着至关重要的作用。尽管PI-PLC活性升高与病理生理状况之间存在良好的联系,但目前还没有PI-PLC抑制剂处于临床开发阶段。此外,现有的抑制剂仅显示有限的效力。目的:dl -1-十二烷基磺酰基-肌醇-3,5-二磷酸被指定为酸性鞘磷脂酶抑制剂,由于其结构与已知的肌醇骨架抑制剂相似,因此应测试其衍生物对PI-PLC活性的抑制作用。方法:研究新合成的化合物对猪血小板裂解液和猪脑匀浆中PI-PLC活性的抑制作用,以及对重组同工酶plc - γ1和plc - γ2的抑制作用。采用高效液相色谱联用质谱法测定L-α-磷脂酰肌醇中二酰基甘油的释放量。此外,通过高效液相色谱法测定所选化合物对其他表面活性剂的抑制效力,从而评估所选化合物的特异性。结果:发现dl -1- o-十二烷基磺酰基肌醇-3,5-二磷酸在微摩尔浓度下抑制PI-PLC活性。然而,其最大可达到的抑制效果被限制在约70%。通过结构修饰,开发了抑制剂,几乎完全抑制了PIPLC的活性。该研究还揭示了所谓的PI-PLC抑制剂U73122,在文献中仍然经常被引用来证明PI-PLC参与生化过程,不适合用于此目的。与其他人的观察结果一致,其对含生物嗜核细胞或组织制剂的抑制活性明显低于其对纯化PI-PLC酶的抑制活性。此外,U73122被证明可以抑制其他酶,如胞质磷脂酶A2α、脂肪酸酰胺水解酶和单酰基甘油脂肪酶,这些酶和PI-PLC一样,代谢亲脂底物。相比之下,与U73122相比,新开发的肌醇衍生物对生物亲核细胞的敏感性降低,对被测表面活性剂的选择性显著提高。结论:已鉴定出对PI-PLC具有明显抑制活性的新化合物。这些发现可能对临床应用的PI-PLC抑制剂的开发有价值,特别是对癌症的治疗。此外,与通常用于科学研究的马来酰亚胺衍生物U73122相比,所开发的肌醇衍生物更适合于研究PI-PLC在组织匀浆中生理过程中的作用。这一优势源于U73122是一种非特异性“泛分析干扰化合物”(PAIN)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Medicinal Chemistry
Medicinal Chemistry 医学-医药化学
CiteScore
4.30
自引率
4.30%
发文量
109
审稿时长
12 months
期刊介绍: Aims & Scope Medicinal Chemistry a peer-reviewed journal, aims to cover all the latest outstanding developments in medicinal chemistry and rational drug design. The journal publishes original research, mini-review articles and guest edited thematic issues covering recent research and developments in the field. Articles are published rapidly by taking full advantage of Internet technology for both the submission and peer review of manuscripts. Medicinal Chemistry is an essential journal for all involved in drug design and discovery.
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