Faculty of Pharmacy, Widya Mandala Catholic University Surabaya, Jalan Kalisari Selatan 1, Surabaya, 60237, East Java, Indonesia.
Institute of Anatomy and Cell Biology, Julius-Maximilians-University Würzburg, Würzburg, Germany.
Prostaglandins Other Lipid Mediat. 2021 Jun;154:106549. doi: 10.1016/j.prostaglandins.2021.106549. Epub 2021 Apr 5.
Salicylic acid derivate is very popular for its activity to suppress pain, fever, and inflammation. One of its derivatives is acetylsalicylic acid (ASA) which has been reported repeatedly that, as a non-steroidal anti-inflammatory drug (NSAID), it has a cardioprotective effect. Although ASA has various advantages, several studies have reported that it may induce severe peptic ulcer disease. We recently synthesized a new compound derived from salicylic acid, namely 2-((3-(chloromethyl)benzoyl)oxy)benzoic acid (3-CHCl) which still has the benefit of acetylsalicylic acid as an analgesic and antiplatelet, but lacks its harmful side effects (Caroline et al., 2019). In addition, in silico studies of 3-CHCl showed a higher affinity towards protein receptor cyclooxygenase-2 (COX-2; PDB: 5F1A) than ASA. We hypothesized that 3-CHCl inhibits the COX-2 activity which could presumably decrease the inflammatory responses. However, no knowledge is available on the anti-inflammatory response and molecular signaling of this new compound. Hence, in this study, we investigated the potential functional relevance of 3-CHCl in regulating the inflammatory response in lipopolysaccharide (LPS)-induced rats. The results of this study show that this compound could significantly reduce the inflammatory parameter in LPS-induced rats.
Rats were induced with LPS of 0.5 mg/kg bw intravenously, prior oral administration with vehicle (3% Pulvis Gummi Arabicum / PGA), 500 mg/60 kg body weight (bw; rat dosage converted to human) of 3-CHCl and ASA. The inflammatory parameters such as changes in the temperature of septic shock, cardiac blood plasma concentrations of IL-1β and TNF-α (ELISA), blood inflammation parameters, white blood cell concentrations, and lung histopathology were observed. Meanwhile, the stability of 3-CHCl powder was evaluated.
After the administration of 500 mg/60 kg bw of 3-CHCl (rat dosage converted to human) to LPS-induced rats, we observed a significant reduction of both TNF-α (5.70+/-1.04 × 10 pg/mL, p=<0.001) and IL-1β (2.32+/-0.28 × 10 pg/mL, p=<0.001) cardiac blood plasma concentrations. Besides, we found a reduction of white blood cell concentration and the severity of lung injury in the 3-CHCl group compared to the LPS-induced rat group. Additionally, this compound maintained the rat body temperature within normal limits during inflammation, preventing the rats to undergo septic shock, characterized by hypothermic (t = 120 min.) or hyperthermic (t = 360 min) conditions. Furthermore, 3-CHCl was found to be stable until 3 years at 25°C with a relative humidity of 75 ± 5%.
3-CHCl compound inhibited inflammation in the LPS-induced inflammation response model in rats, hypothetically through binding to COX-2, and presumably inhibited LPS-induced NF-κβ signaling pathways. This study could be used as a preliminary hint to investigate the target molecular pathways of 3-CHCl as a novel and less toxic therapeutical agent in alleviating the COX-related inflammatory diseases, and most importantly to support the planning and development of clinical trial.
水杨酸衍生物因其具有抑制疼痛、发热和炎症的活性而广受欢迎。其衍生物之一是乙酰水杨酸（ASA），已有多项研究报道称，作为一种非甾体抗炎药（NSAID），它具有心脏保护作用。尽管 ASA 具有多种优势，但已有几项研究报道称，它可能会引起严重的消化性溃疡疾病。我们最近合成了一种源自水杨酸的新化合物，即 2-（（3-（氯甲基）苯甲酰基）氧基）苯甲酸（3-CHCl），它仍然具有 ASA 作为镇痛药和抗血小板的作用，但缺乏其有害的副作用（Caroline 等人，2019 年）。此外，3-CHCl 的计算机模拟研究表明，它与环氧化酶-2（COX-2；PDB：5F1A）的蛋白受体结合的亲和力高于 ASA。我们假设 3-CHCl 抑制 COX-2 活性，这可能会降低炎症反应。然而，对于这种新化合物的抗炎反应和分子信号通路，目前还没有相关知识。因此，在这项研究中，我们研究了 3-CHCl 在调节脂多糖（LPS）诱导的大鼠炎症反应中的潜在功能相关性。研究结果表明，该化合物可显著降低 LPS 诱导的大鼠的炎症参数。
大鼠静脉注射 0.5mg/kg bw 的 LPS，预先口服给予 3%阿拉伯胶（PGA）作为载体、500mg/60kg 体重（大鼠剂量转换为人）的 3-CHCl 和 ASA。观察了败血症休克时体温变化、心脏血血浆中白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）浓度（ELISA）、血液炎症参数、白细胞浓度和肺组织病理学等炎症参数。同时，评估了 3-CHCl 粉末的稳定性。
给 LPS 诱导的大鼠口服 500mg/60kg bw 的 3-CHCl（大鼠剂量转换为人）后，我们观察到 TNF-α（5.70+/-1.04×10pg/mL，p=<0.001）和 IL-1β（2.32+/-0.28×10pg/mL，p=<0.001）心脏血血浆浓度明显降低。此外，与 LPS 诱导的大鼠组相比，3-CHCl 组的白细胞浓度和肺损伤严重程度均有所降低。此外，该化合物在炎症期间将大鼠体温维持在正常范围内，防止大鼠发生败血症性休克，其特征为体温过低（t=120min.）或体温过高（t=360min.）。此外，3-CHCl 在 25°C 下相对湿度为 75±5%时可稳定 3 年。
3-CHCl 化合物抑制了 LPS 诱导的炎症反应模型中的炎症反应，推测是通过与 COX-2 结合，从而抑制 LPS 诱导的 NF-κβ信号通路。这项研究可以作为探索 3-CHCl 作为一种新型、毒性较小的治疗药物在缓解 COX 相关炎症性疾病中的靶向分子通路的初步线索，最重要的是为规划和开展临床试验提供支持。
