Enzymatic synthesis of DHA-lysophosphatidylcholine and evaluation of its effect on the MDA-MB-231 human breast cancer cell line

• Main Author: Mohamad Ali, Dalal
• Other Authors: Biologie des Organismes, Stress, Santé, Environnement Le Mans Université (BiOSSE), Le Mans Université (UM), Le Mans Université, Lionel Ulmann, Gaëlle Pencreac'h, Laurent Poisson
• Format: Doctoral or Postdoctoral Thesis
• Language: English
• Published: HAL CCSD 2022
• Subjects: DHA-Lysophosphatidylcholine; Solvent free esterification; Immobilized lipase; Design of experiments; Response surface methodology; MDA-MB-231 cell line; Cell viability; Cell death mechanisms; Estérification sans solvant; Lipase immobilisée; Plan d'expériences; Méthode de réponse de surface; Lignée cellulaire MDA-MB-231; Viabilité cellulaire; Mécanismes de mort cellulaire; [SDV.BC]Life Sciences [q-bio]/Cellular Biology; [CHIM.THER]Chemical Sciences/Medicinal Chemistry; Antarc*; Antarctica
• Online Access: https://theses.hal.science/tel-03934821; https://theses.hal.science/tel-03934821/document; https://theses.hal.science/tel-03934821/file/2022LEMA1003.pdf

This work studies the synthesis of DHA-lysophosphatidylcholine (LPC-DHA) by solvent-free lipase-catalyzed esterification and the ability of LPC-DHA to reduce cell viability of the human cancer cell line MDA-MB-231. Optimization of esterification parameters between glycerophosphatidylcholine (GPC) and docosahexaenoic acid (DHA) was performed using response surface methodology (RSM). Two responses were measured: the conversion yield of GPC and the concentration of LPC-DHA in the medium. A GPC conversion yield of 67% is obtained under the following reaction conditions: a DHA/GPC molar ratio of 17, a reaction temperature of 36°C and a Novozym® 435 (immobilized lipase B from Candida antarctica) load of 15%. The maximum concentration of LPC-DHA obtained is 245 mM under the following conditions: a DHA/GPC molar ratio of 4, temperature of 36°C and a Novozym® 435 load of 15%. NMR characterization confirmed that the synthesized compound is pure and unoxidized sn-1 LPC-DHA In vitro study of the effect of different LPCs and different lipid vectors of DHA on MDA-MB-231 showed that LPC-DHA was the most effective in reducing cell viability, with IC50 for LPC-DHA, PC-DHA, MAG-DHA, and free DHA of 19 µM, 50 µM, 300 µM, and 347 µM, respectively. LPC-DHA and PC-DHA reduce cell viability primarily by inducing oxidative stress and plasma membrane damage. DHA and MAG-DHA also induced oxidative stress. In conclusion, this work led to the synthesis of LPC-DHA under favorable conditions and demonstrated the very interesting effect of LPC-DHA in reducing the viability of MDA-MB-231 breast cancer cells. Ce travail étudie la synthèse de DHA-lysophosphatidylcholine (LPC-DHA) par estérification enzymatique, catalysée par une lipase et sans solvant, ainsi que la capacité de la LPC-DHA à réduire la viabilité cellulaire de la lignée cancéreuse humaine MDA-MB-231. Une optimisation des paramètres de l’estérification entre la glycerophosphatidylcholine (GPC) et l’acide docosahexaénoïque (DHA, C22 :6 -3) a été réalisée selon la méthodologie des ...