Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, 1200 Cailun Road, Shanghai, 201203, China.
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Compound Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, 1200 Cailun Road, Shanghai, 201203, China.
J Ethnopharmacol. 2022 Oct 28;297:115569. doi: 10.1016/j.jep.2022.115569. Epub 2022 Jul 19.
Asari Radix et Rhizoma (ARR), including 3 major plants of genus Asarum Linn, A. heterotropoides Fr. Schmidt var. mandshuricum (Maxim.) Kitag., A. sieboldii Miq. f. sieboldii and A. sieboldii Miq f. seoulense (Nakai) C. Y. Cheng et C. S. Yang, is one of the most important traditional herbal medicine in Asia with tremendous pharmacological activities. For a long time, researchers focus attention on studing asarinin and essential oils, the indicating ingredients of ARR, but paid less attention to another characteristic component, alkamides. The role of alkamides in the major efficacy of ARR medication remains to be elucidated.
This study aims to investigate the contribution of alkamides in the efficacy of ARR according to the evaluation of antinociceptive and anti-inflammatory effects and in vivo pharmacokinetics processes.
For pharmacodynamic study, the analgesic and anti-inflammatory effects of alkamides-enriched fraction (ARRA) were comparatively evaluated by writhing test, hot plate test, and ear swelling test in mice after oral administration. For pharmacokinetic study, an UHPLC-MS/MS method was developed for the simultaneous determination of N-isobutyl-2E,4E,8Z,10Z/E-dodecatetraenamide (DDA) and other 6 major characteristic ingredients of ARR in rat plasma. The analytical method was validated and successfully applied to the pharmacokinetic study of ARR extract and DDA.
Pharmacodynamic study show that the ARR and ARRA can significantly inhibit the writhing times of mice caused by acetic acid administration, increase the pain threshold of thermal stimulation, and inhibit xylene treated ear swelling degree by reduce PGE and TNF-α levels in the inflamed tissue. For pharmacokinetic study, the pharmacokinetic parameters of Vd/F and CL/F after intravenous administration in rats of DDA are 63.94 ± 32.12 L/kg and 0.33 ± 0.06 L/min/kg, respectively. The plasma drug concentration declined with the T value of 2.25 ± 0.96 h, and the MRT was 2.23 ± 1.02 h. The absolute bioavailability of DDA after oral administration was calculated as 10.73%. DDA, methyleugenol, and asarinin have relatively high AUC values when the ethanol and water extract of ARR is orally administered.
ARRA is a kind of active ingredients with potential analgesic and anti-inflammatory effects that played a significant role in the major efficacy of ARR. DDA, the major compound of ARRA, has a high level of exposure in vivo, which could be is suitable for the pharmacokinetic marker or new quality marker of ARR.
细辛根和根茎（ARR），包括细辛属的 3 种主要植物，即细辛 Asarum heterotropoides Fr. Schmidt var. mandshuricum（Maxim.）Kitag.、北细辛 A. sieboldii Miq. f. sieboldii 和汉城细辛 A. sieboldii Miq. f. seoulense（Nakai）C. Y. Cheng et C. S. Yang，是亚洲最重要的传统草药之一，具有巨大的药理学活性。长期以来，研究人员一直关注细辛素和精油，这是 ARR 的指示成分，但对另一种特征成分——酰胺类化合物关注较少。酰胺类化合物在 ARR 药物主要疗效中的作用仍有待阐明。
本研究旨在通过口服后扭体试验、热板试验和耳肿胀试验，评估酰胺类化合物在 ARR 疗效中的作用，评价其镇痛和抗炎作用及体内药代动力学过程。
在药效学研究中，比较酰胺类化合物富集部分（ARRA）对乙酸给药引起的小鼠扭体次数、热刺激痛阈和二甲苯处理耳肿胀程度的影响。在药代动力学研究中，建立了一种同时测定大鼠血浆中 N-异丁基-2E,4E,8Z,10Z/E-十二碳四烯酰胺（DDA）和 ARR 其他 6 种主要特征成分的 UHPLC-MS/MS 方法。对分析方法进行了验证，并成功应用于 ARR 提取物和 DDA 的药代动力学研究。
药效学研究表明，ARR 和 ARRA 能显著抑制醋酸致小鼠扭体次数，提高热刺激痛阈，降低炎性组织中 PGE 和 TNF-α水平，抑制二甲苯致耳肿胀程度。药代动力学研究结果表明，DDA 静脉给药后大鼠的 Vd/F 和 CL/F 药代动力学参数分别为 63.94±32.12 L/kg 和 0.33±0.06 L/min/kg。DDA 的血浆药物浓度随 T 值的变化而下降，MRT 为 2.23±1.02 h。DDA 口服绝对生物利用度为 10.73%。当 ARR 的乙醇和水提取物口服时，DDA、甲基丁香酚和细辛素具有相对较高的 AUC 值。
ARRA 是一种具有潜在镇痛和抗炎作用的活性成分，在 ARR 的主要疗效中发挥了重要作用。DDA 作为 ARRA 的主要化合物，在体内具有较高的暴露水平，可作为 ARR 的药效标志物或新的质量标志物。
