Abstract：
Gamma-terpinene (γ-TPN) is a cyclohexane monoterpene isolated from plant essential oils, such as tea tree (Melaleuca alternifolia), oregano (Origanum vulgare), rosemary (Rosmarinus officinalis L.), thyme (Thymus vulgaris Marchand), and eucalyptus (Eucalyptus sp.). Terpenes are widely studied molecules pharmacologically active on the cardiovascular system, hemostasis, and antioxidant actions. Herein, it was investigated the cytotoxic and antiplatelet activity of γ-TPN using different non-clinical laboratory models. For in silico evaluation, the PreADMET, SwissADME, and SwissTargetPrediction softwares were used. Molecular docking was performed using the AutoDockVina and BIOVIA Discovery Studio databases. The cytotoxicity of γ-TPN was analyzed by the MTT assay upon normal murine endothelial SVEC4-10 and fibroblast L-929 cells. Platelet aggregation was evaluated with platelet-rich (PRP) and platelet-poor (PPP) plasma from spontaneously hypertensive rats (SHR), in addition to SVEC4-10 cells pre-incubated with γ-TPN (50, 100, and 200 µM) for 24 h. SHR animals were pre-treated by gavage with γ-TPN for 7 days and divided into four groups (negative control, 25, 50, and 100 mg/kg). Blood samples were collected to measure nitrite using the Griess reagent. Gamma-TPN proved to be quite lipid-soluble (Log P = +4.50), with a qualified profile of similarity to the drug, good bioavailability, and adequate pharmacokinetics. It exhibited affinity mainly for the P2Y12 receptor (6.450 ± 0.232 Kcal/mol), moderate cytotoxicity for L-929 (CC50 = 333.3 µM) and SVEC 4-10 (CC50 = 366.7 µM) cells. The presence of γ-TPN in SVEC 4-10 cells was also able to reduce platelet aggregation by 51.57 and 44.20% at lower concentrations (50 and 100 µM, respectively). Then, γ-TPN has good affinity with purinergic receptors and an effect on the reversal of platelet aggregation and oxidative stress, being promising and safe for therapeutic targets and subsequent studies on the control of thromboembolic diseases.
摘要：
γ-萜品烯（γ-TPN）是从植物精油中分离出的环己烷单萜，如茶树（互叶白千层），牛至(Ohoganumvulgare),迷迭香（迷迭香），百里香，和桉树(桉树).萜烯是广泛研究的分子，对心血管系统具有药理活性，止血,和抗氧化作用。在这里，使用不同的非临床实验室模型研究了γ-TPN的细胞毒性和抗血小板活性.对于计算机评估，PreADMET,Swissadme,并使用SwissTargetPrediction软件。使用AutoDockVina和BIOVIADiscoveryStudio数据库进行分子对接。通过MTT测定法分析γ-TPN对正常鼠内皮SVEC4-10和成纤维细胞L-929细胞的细胞毒性。使用来自自发性高血压大鼠（SHR）的富血小板（PRP）和贫血小板（PPP）血浆评估血小板聚集，除了与γ-TPN（50、100和200µM）预孵育24小时的SVEC4-10细胞外。SHR动物通过用γ-TPN管饲法预处理7天，并分为四组（阴性对照，25、50和100mg/kg)。收集血样以使用Griess试剂测量亚硝酸盐。γ-TPN被证明是相当脂溶性的（LogP=+4.50），具有与药物相似的合格概况，良好的生物利用度，和足够的药代动力学。主要对P2Y12受体有亲和力(6.450±0.232Kcal/mol),L-929(CC50=333.3µM)和SVEC4-10(CC50=366.7µM)细胞的中等细胞毒性。SVEC4-10细胞中γ-TPN的存在也能够在较低浓度（50和100μM，分别)。然后，γ-TPN与嘌呤受体具有良好的亲和力，对血小板聚集和氧化应激的逆转有作用，有希望的和安全的治疗目标和随后的研究控制血栓栓塞性疾病。
