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.

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.

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.

Pre-meal high-performance inulin supplementation reduce post-prandial glycaemic response in healthy subjects: A repeated single-arm clinical trial.

BACKGROUND AND AIMS: High-performance (HP) inulin, a dietary fiber consists of more than 10 fructose polymers, have been shown to reduce post-prandial glycaemic response (PPGR) and could prevent the occurrence of Type-2 diabetes mellitus (T2DM). Currently, there are no data on whether pre-meal HP inulin supplementation could decrease PPGR. METHODS: 8 healthy adults consumed 20 g of formula that contain 60.2% inulin (w/w) dissolved in water. Blood glucose was measured in fasted participants and at 30-120 min after starting to eat a prepared meal. This test was repeated every week with different supplement formulas. CONCLUSION: pre-meal HP Inulin formula supplementation could suppress the post-prandial glycaemic response.

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.

The combination of isomalto-oligosaccharides (IMO)-based dietary fiber and hypocaloric high-protein diet could improve the anthropometric profile and fasting plasma glucose of healthy adults: A repeated single-arm clinical trial.

Background and aims: Meals with high protein and fiber could reduce weight and improve diabetes risk factors. Isomalto-oligosaccharide (IMO), a form of dietary fiber, could induce the afferent signal that causes appetite suppression. However, the direct effect of fiber supplementation in the form of IMO combined with a high-protein diet (HPF) on those parameters is still unknown. This study aims to investigate the effect of HPF on anthropometric parameters and blood glucose regulation of healthy subjects. . Methods: Thirteen healthy subjects were given a hypocaloric high protein diet (HPD) mixed with their prepared meals for two weeks. Followed by the HPF diet for another two weeks. Their anthropometric parameters, such as body composition (total body weight, body fat percentage, and fat-free mass), BMI and waist circumference, and fasting plasma glucose, were measured. Results: Compared to pre-intervention, HPF could significantly (p ≤ 0.004) reduce the anthropometric parameters and fasting plasma glucose. Compared to HPD, HPF could significantly (p ≤ 0.005) reduce more total body weight, body fat percentage, and BMI. In addition, HPF could induce more satiety than HPD (higher VAS score). Conclusion: HPF could improve the subject's anthropometric parameters which is obviously beneficial in preventing the risk of developing diabetes.