Olga V. Oskolkova , Bernd Gesslbauer , Valery Bochkov
{"title":"烷基酰胺支架上氧化磷脂的简化合成","authors":"Olga V. Oskolkova , Bernd Gesslbauer , Valery Bochkov","doi":"10.1016/j.mex.2025.103288","DOIUrl":null,"url":null,"abstract":"<div><div>Oxidized phospholipids (OxPLs), containing oxidized fatty acids (oxylipins), play a significant role in various diseases. However, studying the structure-activity relationships of OxPLs and their signaling mechanisms is challenging due to the complexity of the chemical synthesis of structurally distinct lipid species. In this study, we aimed to develop a method for attaching free oxylipins to a lysophospholipid to form OxPLs. We hypothesized that oxylipins could be conjugated to PLs <em>via</em> a known chemical reaction between activated esters of carboxylic acids and amino groups. The carboxyl groups of oxylipins were activated using N-hydroxysuccinimide and a coupling reagent, then conjugated to a lyso-phosphatidylcholine containing NH<sub>2</sub>-groupd at <em>sn</em>-2 position, forming amide bonds. All reactions were performed under mild conditions and demonstrated high yields. To prevent acyl migration, the <em>sn</em>-1 position of PLs was modified with an alkyl residue linked <em>via</em> an ether bond. Several oxylipin-containing PLs were successfully synthesized, isolated, and characterized. The anti-TLR4 and endothelial barrier-protective activities of these alkyl-amide OxPLs were found to be equivalent to diacyl-OxPLs. This method enables efficient synthesis of modified OxPLs for biological testing. The combination of ether and amide bonds enhances biological stability and simplifies effect analysis.<ul><li><span>•</span><span><div>The method describes the preparation of a single precursor for multiple choline PLs, specifically 2-deoxy-2-amino-1-lyso-<em>sn</em>-3-glycerophosphocholine, followed by the attachment of oxylipins to it.</div></span></li><li><span>•</span><span><div>No protection-deprotection steps are needed for oxylipins for the synthesis of phosphatidylcholines.</div></span></li><li><span>•</span><span><div>Isolation of compounds is performed using fast liquid-liquid and solid-phase extractions.</div></span></li></ul></div></div>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"14 ","pages":"Article 103288"},"PeriodicalIF":1.6000,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Simplified synthesis of oxidized phospholipids on alkyl-amide scaffold\",\"authors\":\"Olga V. Oskolkova , Bernd Gesslbauer , Valery Bochkov\",\"doi\":\"10.1016/j.mex.2025.103288\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>Oxidized phospholipids (OxPLs), containing oxidized fatty acids (oxylipins), play a significant role in various diseases. However, studying the structure-activity relationships of OxPLs and their signaling mechanisms is challenging due to the complexity of the chemical synthesis of structurally distinct lipid species. In this study, we aimed to develop a method for attaching free oxylipins to a lysophospholipid to form OxPLs. We hypothesized that oxylipins could be conjugated to PLs <em>via</em> a known chemical reaction between activated esters of carboxylic acids and amino groups. The carboxyl groups of oxylipins were activated using N-hydroxysuccinimide and a coupling reagent, then conjugated to a lyso-phosphatidylcholine containing NH<sub>2</sub>-groupd at <em>sn</em>-2 position, forming amide bonds. All reactions were performed under mild conditions and demonstrated high yields. To prevent acyl migration, the <em>sn</em>-1 position of PLs was modified with an alkyl residue linked <em>via</em> an ether bond. Several oxylipin-containing PLs were successfully synthesized, isolated, and characterized. The anti-TLR4 and endothelial barrier-protective activities of these alkyl-amide OxPLs were found to be equivalent to diacyl-OxPLs. This method enables efficient synthesis of modified OxPLs for biological testing. The combination of ether and amide bonds enhances biological stability and simplifies effect analysis.<ul><li><span>•</span><span><div>The method describes the preparation of a single precursor for multiple choline PLs, specifically 2-deoxy-2-amino-1-lyso-<em>sn</em>-3-glycerophosphocholine, followed by the attachment of oxylipins to it.</div></span></li><li><span>•</span><span><div>No protection-deprotection steps are needed for oxylipins for the synthesis of phosphatidylcholines.</div></span></li><li><span>•</span><span><div>Isolation of compounds is performed using fast liquid-liquid and solid-phase extractions.</div></span></li></ul></div></div>\",\"PeriodicalId\":18446,\"journal\":{\"name\":\"MethodsX\",\"volume\":\"14 \",\"pages\":\"Article 103288\"},\"PeriodicalIF\":1.6000,\"publicationDate\":\"2025-03-27\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"MethodsX\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2215016125001347\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MULTIDISCIPLINARY SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"MethodsX","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2215016125001347","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MULTIDISCIPLINARY SCIENCES","Score":null,"Total":0}
Simplified synthesis of oxidized phospholipids on alkyl-amide scaffold
Oxidized phospholipids (OxPLs), containing oxidized fatty acids (oxylipins), play a significant role in various diseases. However, studying the structure-activity relationships of OxPLs and their signaling mechanisms is challenging due to the complexity of the chemical synthesis of structurally distinct lipid species. In this study, we aimed to develop a method for attaching free oxylipins to a lysophospholipid to form OxPLs. We hypothesized that oxylipins could be conjugated to PLs via a known chemical reaction between activated esters of carboxylic acids and amino groups. The carboxyl groups of oxylipins were activated using N-hydroxysuccinimide and a coupling reagent, then conjugated to a lyso-phosphatidylcholine containing NH2-groupd at sn-2 position, forming amide bonds. All reactions were performed under mild conditions and demonstrated high yields. To prevent acyl migration, the sn-1 position of PLs was modified with an alkyl residue linked via an ether bond. Several oxylipin-containing PLs were successfully synthesized, isolated, and characterized. The anti-TLR4 and endothelial barrier-protective activities of these alkyl-amide OxPLs were found to be equivalent to diacyl-OxPLs. This method enables efficient synthesis of modified OxPLs for biological testing. The combination of ether and amide bonds enhances biological stability and simplifies effect analysis.
•
The method describes the preparation of a single precursor for multiple choline PLs, specifically 2-deoxy-2-amino-1-lyso-sn-3-glycerophosphocholine, followed by the attachment of oxylipins to it.
•
No protection-deprotection steps are needed for oxylipins for the synthesis of phosphatidylcholines.
•
Isolation of compounds is performed using fast liquid-liquid and solid-phase extractions.