Levomefolic acid is an orally available, bioactive form of folic acid that crosses the blood-brain barrier[1]. Levomefolic acid, a ligand with low binding energy, can form three hydrogen bonds with Arg169, Asp305 and Tyr276 of COVID-19 PLpro protein and form π-π stacking interaction with Tyr271[2]. Levomefolic acid has been widely used as a model compound and an internal reference to develop novel LC-MS/MS methods for the identification of related compounds[3].
In vitro, Levomefolic acid treatment at 10µg/ml for 24 hours significantly inhibited Caco-2 cell proliferation and induced cell cycle arrest[4]. Treatment of BEAS-2B cells with 0.012g/L Levomefolic acid for 24h protected BEAS-2B cells from oxidative stress and inflammatory injury induced by glucose (50mM) and restored cell viability[5]. Treatment with 50µM Levomefolic acid for 48h significantly induced DNA damage in MRC5 and IMR90 cells, resulting in decreased MTR gene expression and increased MTHFD1, EGFR, XRCC1, and DNA Polβ gene expression[6].
In vivo, intraperitoneal injection of Levomefolic acid (3µg/kg) at 30 minutes before ischemia and again 3 hours after reperfusion can improve renal function in Sprague-Dawley rats, alleviate oxidative stress, restore glutathione levels in the kidneys, and reduce lipid peroxidation[7]. Within 5 weeks after weaning, oral administration of 4ml of drinking water containing a dose of 47μg/ml of Levomefolic acid daily significantly increased the body weight of female mice and promoted the maturation of oocytes[8].
References:
[1] Lam N S K, Long X X, Li X, et al. The potential use of folate and its derivatives in treating psychiatric disorders: A systematic review[J]. Biomedicine & Pharmacotherapy, 2022, 146: 112541.
[2] Hosseini M, Chen W, Xiao D, et al. Computational molecular docking and virtual screening revealed promising SARS-CoV-2 drugs[J]. Precision clinical medicine, 2021, 4(1): 1-16.
[3] Golja M V, Trontelj J, Geršak K, et al. Simultaneous quantification of intracellular concentrations of clinically important metabolites of folate-homocysteine cycle by LC-MS/MS[J]. Analytical biochemistry, 2020, 605: 113830.
[4] Akoglu B, Milovic V, Caspary W F, et al. Hyperproliferation of homocysteinetreated colon cancer cells is reversed by folate and 5-methyltetrahydrofolate[J]. European journal of Nutrition, 2004, 43(2): 93-99.
[5] Pathikkal A, Puthusseri B, Divya P, et al. Folate derivatives, 5-methyltetrahydrofolate and 10-formyltetrahydrofolate, protect BEAS-2B cells from high glucose–induced oxidative stress and inflammation[J]. In Vitro Cellular & Developmental Biology-Animal, 2022, 58(5): 419-428.
[6] Karkera C, Senejani A G. Evaluating toxicity and level of DNA damage in human fetal lung cells upon exposure to 5-methyltetrahydrofolate (bioactive folate)[J]. Nutrition and Health, 2024: 02601060241302895.
[7] Wijerathne C U B, Au-Yeung K K W, Siow Y L, et al. 5-methyltetrahydrofolate attenuates oxidative stress and improves kidney function in acute kidney injury through activation of Nrf2 and antioxidant defense[J]. Antioxidants, 2022, 11(6): 1046.
[8] Nasr‐Esfahani F, Jafarpour F, Varnosfaderani S R, et al. Differential Effects of Folic Acid and 5‐Methyltetrahydrofolate on Mice Oocytes and Embryo Development In Vitro and In Vivo: Implications for Reproductive Health[J]. The FASEB Journal, 2025, 39(20): e71090.
Levomefolic acid是一种具有口服活性且可通过血脑屏障的叶酸生物活性形式[1]。Levomefolic acid能以低结合能作为配体，与COVID-19 PLpro蛋白的Arg169、Asp305和Tyr276形成三个氢键，并与Tyr271产生π-π堆叠相互作用[2]。Levomefolic acid已作为模型化合物和内参物广泛应用于开发鉴定相关化合物的新型LC-MS/MS方法[3]。
在体外，使用10µg/ml的Levomefolic acid处理Caco-2细胞24小时，可显著抑制Caco-2细胞增殖并诱导细胞周期阻滞[4]。用0.012g/L的Levomefolic acid处理BEAS-2B细胞24小时，能保护细胞免受葡萄糖(50mM)诱导的氧化应激和炎症损伤，并恢复细胞活力[5]。以50µM的Levomefolic acid处理MRC5和IMR90细胞48小时，可显著诱导DNA损伤，导致MTR基因表达下调，同时上调MTHFD1、EGFR、XRCC1和DNA Polβ基因表达[6]。
在体内，在缺血前30分钟及再灌注后3小时分别腹腔注射3µg/kg剂量的Levomefolic acid，可改善Sprague-Dawley大鼠的肾功能，减轻氧化应激，恢复肾脏谷胱甘肽水平并降低脂质过氧化[7]。在小鼠断奶后5周内每日口服4ml含47μg/ml的Levomefolic acid的饮用水，可显著增加雌鼠体重并促进卵母细胞成熟[8]。