2-(3-(Chloromethyl)benzoyloxy)benzoic Acid Reduces Prostaglandin E-2 Concentration, NOX2 and NFKB Expression, ROS Production, and COX-2 Expression in Lipopolysaccharide-Induced Mice.

INTRODUCTION: Inflammation is a fundamental response to various insults, including microbial invasion and tissue injury. While aspirin (ASA) has been widely used for its anti-inflammatory properties, its adverse effects and limitations highlight the need for novel therapeutic alternatives. Recently, a novel salicylic acid derivative, 2-((3-(chloromethyl)benzoyl)oxy)benzoic acid (3-CH2Cl), has emerged as a potential substitute for ASA, offering a simpler, environmentally friendly synthesis and a promising safety profile. AIM OF THE STUDY: This research aims to evaluate the anti-inflammatory mechanism of 3-CH2Cl in a lipopolysaccharide (LPS)-induced mouse model, focusing on its effects on prostaglandin E-2 (PGE-2) concentration, NOX2 and NFkB expression, ROS production, and COX-2 expression. MATERIAL AND METHODS: Utilizing BALB/C mice subjected to LPS-induced inflammation, we investigated the therapeutic potential of 3-CH2Cl. The study included synthesis and tablet preparation, experimental design, peripheral blood plasma PGE-2 measurement, splenocyte isolation and COX-2 expression analysis, nitric oxide and ROS measurement, and immunohistochemical analysis of NOX2 and NFkB expression. RESULTS: 3-CH2Cl significantly reduced PGE-2 levels (p=0.005), NO concentration in liver homogenates (p=0.005) and plasma (p=0.0011), and expression of NOX2 and NFkB in liver (p<0.0001) and splenocytes (p=0.0036), demonstrating superior anti-inflammatory activity compared to ASA. Additionally, it showed potential in decreasing COX-2 expression in splenocytes. CONCLUSION: 3-CH2Cl exhibits potent anti-inflammatory properties, outperforming ASA in several key inflammatory markers in an LPS-induced inflammation model. The reduction of COX-2 expression, alongside the reduction of pro-inflammatory cytokines and oxidative stress markers, suggest it as a promising therapeutic agent for various inflammatory conditions.

Synthesis, Characterization, and Application of 2-((3-(Chloromethyl)-benzoyl)oxy)benzoic Acid: A Review.

Salicylic acid (SA) derivate is well-known for its anti-inflammatory and analgesic activity through cyclooxygenase (COX)-inhibition. Previous studies pointed toward gastric toxicity induced by most salicylic acid derivative compounds, particularly acetylsalicylic acid (ASA). Despite the adverse effect, ASA is still used due to price affordability and additional advantages in preventing platelet aggregation. Recently, a novel salicylic acid derivative called 2-((3 (chloromethyl)benzoyl)oxy)benzoic acid (3-CH2Cl) was introduced as a potential alternative compound to substitute ASA. Preliminary assessment results of COX-2 specificity, toxicity profile, analgesic, anti-inflammatory, and antiplatelet activity have made 3-CH2Cl a promising compound for "new" drug development. This review focuses on the discovery, potential activity, and benefits of 3-CH2Cl and the possible molecular mechanisms of its regulations in health and disease. Thus, this review may prove to be beneficial for the utilization of 3-CH2Cl as a potential alternative drug to substitute ASA.

Tablet Formulation of 2-((3-(Chloromethyl)benzoyl)oxy)benzoic Acid by Linear and Quadratic Models.

PURPOSE: This research determines the effect of sodium lauryl sulfate (SLS) as a surfactant, croscarmellose sodium (CS) as a disintegrating agent, and SLS-CS combinations on 2-((3-(chloromethyl)benzoyl)oxy)benzoic acid (3CH2Cl) (log P = 3.73) tablet formulations. In addition, this study aims to determine the optimum of the 3CH2Cl tablet formula. METHODS: The tablets are manufactured through direct compression according to the simplex lattice design. The optimal SLS and CS concentration was determined in vitro using linear and quadratic models to achieve better tablet disintegration and dissolution. RESULTS: The same linear and quadratic coefficient profiles of SLS and CS indicate that the combined coefficient of SLS-CS with a quadratic model can be used to predict the effect of the SLS-CS combination. Based on the linear model coefficients, SLS and CS increase the value of flow time (9.35; 7.65), Carr index (26.17; 21.17), hardness (9.84; 7.44), friability (0.38; 0.31), disintegrating time (5.74; 2.62), and drug release (84.28; 58.65). The quadratic model coefficient shows that SLS-CS combinations increase flow time (0.60), Carr index (2.00), hardness (1.00), and disintegrating time (1.04). Meanwhile, they decrease friability (-0.02) and drug release (-9.10). CONCLUSIONS: SLS, CS, and SLS-CS combinations affect the quality of tablet mass and tablets. The optimum tablet formula was 3CH2Cl (300 mg), Ne (9.38%), SLS (0.92%), CS (2.33%), MCC (5%), and SDL (ad 800 mg). 3CH2Cl has analgesic activity despite the presence of tablet excipients. The 3CH2Cl tablet is an innovative formulation and a new alternative for future analgesic drugs.

Anti-inflammatory activity of 2-((3-(chloromethyl)benzoyl)oxy)benzoic acid in LPS-induced rat model.

INTRODUCTION: 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-CH2Cl) 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-CH2Cl showed a higher affinity towards protein receptor cyclooxygenase-2 (COX-2; PDB: 5F1A) than ASA. We hypothesized that 3-CH2Cl 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-CH2Cl 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. MATERIAL AND METHODS: 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-CH2Cl and ASA. The inflammatory parameters such as changes in the temperature of septic shock, cardiac blood plasma concentrations of IL-1ß and TNF-α (ELISA), blood inflammation parameters, white blood cell concentrations, and lung histopathology were observed. Meanwhile, the stability of 3-CH2Cl powder was evaluated. RESULT: After the administration of 500 mg/60 kg bw of 3-CH2Cl (rat dosage converted to human) to LPS-induced rats, we observed a significant reduction of both TNF-α (5.70+/-1.04 × 103 pg/mL, p=<0.001) and IL-1ß (2.32+/-0.28 × 103 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-CH2Cl 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-CH2Cl was found to be stable until 3 years at 25°C with a relative humidity of 75 ± 5%. CONCLUSION: 3-CH2Cl compound inhibited inflammation in the LPS-induced inflammation response model in rats, hypothetically through binding to COX-2, and presumably inhibited LPS-induced NF-κß signaling pathways. This study could be used as a preliminary hint to investigate the target molecular pathways of 3-CH2Cl 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.

Phytochemical screening and preliminary clinical trials of the aqueous extract mixture of Andrographis paniculata (Burm. f.) Wall. ex Nees and Syzygium polyanthum (Wight.) Walp leaves in metformin treated patients with type 2 diabetes.

BACKGROUND: Our previous preclinical study showed that the extract mixture (EM) of Andrographis paniculata (Burm. f.) Wall. ex Nees (AP) and Syzygium polyanthum (Wight.) Walp (SP) leaves had antidiabetic effects and were beneficial on alloxan-induced diabetic rats. PURPOSE: The objectives of this study were to: 1) identify the phytochemical compounds present in aqueous extract of AP and SP and 2) examine the benefits of the EM of AP and SP leaves in lowering blood glucose in the presence of standard antidiabetic treatment using metformin in type 2 diabetic patients in Indonesian Traditional Medicine Polyclinic of Dr. Soetomo General Hospital in Surabaya. METHODS: Phytochemical analysis of aqueous leaf extract of AP and SP was performed using standard chemical tests, TLC, and GC-MS. Furthermore, a total of 54 subjects with T2DM participated in this study and were randomly assigned to either the intervention group supplemented with the extract mixture of AP and SP at a dose 900  mg/day for 8 weeks, or the control group which received placebo tablets in a randomized placebo-controlled double-blinded parallel clinical trial. Both groups received metformin at dose 1000  mg/day. Body weight, blood pressure, fasting blood glucose, postprandial glucose, haemoglobin A1c, triglycerides, total cholesterol, low density lipoprotein, high density lipoprotein, and markers of liver and kidney damage were measured. RESULTS: The results of phytochemical analysis showed that the glycosides, terpenoids, alkaloids, flavonoids, saponins, and tannins were found to be present in the extract mixture. GC-MS analyses of AP and SP showed the presence of 19 and 12 peaks, respectively. Methyl ester of 9-octadecenoic and eicosanoic acid were determined as the main constituents of both species. Moreover, the results of clinical study suggested that the extract mixture improved the decrease of fasting blood glucose and postprandial glucose, significantly lowered body mass index compared with the control group. The EM appeared beneficial for SGPT values and uric acid levels. CONCLUSION: Overall, the results of this study suggested the potential beneficial effects of the extract mixture for use as complementary medicine alongside conventional treatment of metformin. The extract mixture contained many highly potent compounds for treating T2DM and preventing short- and long-term risk complications of diabetes.