Villegas-Sánchez Eder, Macías-Alonso Mariana, Osegueda-Robles Soraya, Herrera-Isidrón Lisset, Nuñez-Palenius Hector, González-Marrero Joaquín
Instituto Politécnico Nacional, UPIIG, Av. Mineral de Valenciana, No. 200, Col. Fracc, Industrial Puerto Interior, Silao 36275, Mexico.
División de Ciencias de la Vida, Campus Irapuato-Salamanca, Universidad de Guanajuato, Ex-Hacienda El Copal. Km. 9 Carr. Irapuato-Silao, Irapuato 36824, Mexico.
Pharmaceuticals (Basel). 2021 Jul 29;14(8):747. doi: 10.3390/ph14080747.
Emerging infectious diseases have become a major global problem with public health and economic consequences. It is an urgent need to develop new anti-infective therapies. The natural diterpene carnosol exhibit a wide variety of interesting antibacterial and antiviral properties, and it is considered a theoretical inhibitor of COVID-19 M. However, this compound is present in the family Lamiaceae in low quantities. To obtain carnosol in concentrations high enough to develop pharmacological studies, we evaluated the efficiency of a micropropagation protocol of using a solid medium and a temporary immersion system (TIS), as well as the effect of 6-benzylaminopurine (6-BAP) and α-naphthaleneacetic acid (NAA) on the growth of shoots. Moreover, we developed and validated an analytical method to quantify carnosol using the H-point standard additions method in the high-performance liquid chromatography diode array detector (HPLC-DAD). After 30 days of culture, TIS produced the maximum number of shoots per explant (24.33 ± 1.15) on a liquid medium supplemented with 6-BAP at 5.0 mg L. Next, we also evaluated the effect of immersion time and frequency for TIS. After 72 days of culture, the best results were obtained with an immersion cycle of 1 min every 12 h, yielding 170.33 ± 29.40 shoots. The quantification of carnosol on the samples was performed at a flow rate of 1.2 mL min using binary isocratic mobile phase system 60:40 (/) 10 mM formic acid (pH 3.0) (A) and acetonitrile (B) on a reverse-phase column. The content of carnosol in the in vitro cultures was around 8-fold higher than in the wild plant. The present study represents an efficient alternative method to obtain carnosol for its pre-clinical and clinical development.
新发传染病已成为一个具有公共卫生和经济影响的重大全球问题。开发新的抗感染疗法迫在眉睫。天然二萜类化合物鼠尾草酸具有多种有趣的抗菌和抗病毒特性，被认为是新冠病毒M的理论抑制剂。然而，这种化合物在唇形科植物中含量较低。为了获得足够高浓度的鼠尾草酸以开展药理学研究，我们评估了使用固体培养基和临时浸没系统（TIS）的微繁殖方案的效率，以及6-苄基腺嘌呤（6-BAP）和α-萘乙酸（NAA）对芽生长的影响。此外，我们开发并验证了一种在高效液相色谱二极管阵列检测器（HPLC-DAD）中使用H点标准加入法对鼠尾草酸进行定量的分析方法。培养30天后，TIS在添加了5.0 mg/L 6-BAP的液体培养基上产生的每个外植体芽数最多（24.33±1.15）。接下来，我们还评估了TIS的浸没时间和频率的影响。培养72天后，每12小时浸没1分钟的循环获得了最佳结果，产生了170.33±29.40个芽。在反相柱上，使用二元等度流动相系统60:40（/）10 mM甲酸（pH 3.0）（A）和乙腈（B），以1.2 mL/min的流速对样品中的鼠尾草酸进行定量。体外培养物中鼠尾草酸的含量比野生植物中的高约8倍。本研究为获得用于临床前和临床开发的鼠尾草酸提供了一种有效的替代方法。
