In vitro inhibition potency of malononitrile derivatives on the activity of two pentose phosphate pathway enzymes: accompanied by molecular docking evaluation.

Many disorders, including cancer and malaria, could be targeted via the pentose phosphate pathway (PPP), whose products are key in biosynthetic reactions in cells. The goal of this study was to find new PPP inhibitors. The inhibition effects of malononitrile derivatives on Glucose 6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) were analyzed through in vitro experiments. Besides, molecular docking studies were performed to predict the interactions having role in inhibition of compounds. K i constants of derivatives were found between 4.24 ± 0.46-69.63 ± 7.75 µM for G6PD and 1.91 ± 0.12-95.07 ± 11.08 µM for 6PGD. Derivatives indicated non-competitive inhibition on both enzymes except for compound 4. The findings of the molecular docking studies revealed that free-binding energy estimations agreed with in vitro data. The structure of these malononitrile derivatives may guide for drug discovery in targeting the PPP.

In vitro Inhibition Profiles and Molecular Docking Analysis of Benzohydrazide Derivatives on Red Blood Cell Carbonic Anhydrases Isozymes.

BACKGROUND: Carbonic anhydrases (CAs, EC 4.2.1.1) are metalloenzymes that contain zinc ions on the active side and convert carbon dioxide to bicarbonate in metabolism. Human CA-I and CA-II, which are the most abundant CA isozymes in erythrocytes, have been therapeutic targets in the treatment of glaucoma, hypertension, ulcer, osteoporosis, and, neurological disorders. Benzohydrazides are biologically active compounds, and their various pharmacological effects have been reported. AIM: In light of this, the objective of this study was to investigate the in vitro effects of benzohydrazide derivatives on the activities of hCA-I and hCA-II, determine the compounds as selective inhibitors for these isoenzymes, and estimate the inhibition mechanism through molecular docking studies. METHODS: In this work, we synthesized the 10 different derivatives of benzohydrazide containing various functional group of different positions. RESULTS: As a result, all benzohydrazide derivatives inhibited both isozymes in vitro and 2-amino 3- nitro benzohydrazide (10) was found to be the most efficient inhibitor of both hCA isozymes with the IC50 values of 0.030 and 0.047 µM, respectively. In the molecular docking studies, 3-amino 2- methyl benzohydrazide (3) had the lowest estimated free binding energies against hCA isozymes as -6.43 and -6.13 kcal/mol. CONCLUSION: In this study, hCA-I & II isozymes were isolate from human erythrocytes. CA isozymes are one of these target enzymes. WBC hope that the benzohydrazide derivatives, can guide remedies targeting carbonic anhydrase.

Design and Synthesis of Novel Dual Cholinesterase Inhibitors: In Vitro Inhibition Studies Supported with Molecular Docking.

The major cholinesterase enzymes, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), are important in the therapy of Alzheimer's disease (AD) based on the cholinergic hypothesis. As a result, in recent years, the investigation of dual cholinesterase inhibition methods has become important among scientists. In this study, novel N-(4-chlorobenzyl)-3,4-dimethoxy-N-(m-substituted)benzamide derivatives were synthesized. Then, inhibitory properties of these derivatives were examined in human AChE and BuChE in vitro and possible interactions were determined by molecular docking studies. All benzamide derivatives were exhibited dual inhibitory character and high BBB permeability. The most effective inhibitor was found as N7 for both AChE and BuChE with IC50 values of 1.57 and 2.85 µM, respectively. Besides the most potent inhibitor was predicted as N7 in terms of binding energies with -12.18 kcal/mol and -9.92 kcal/mol, respectively. The reason for these results is that bromine (N7) is the bulkiest molecule among the other substituted groups. These derivatives could be exploited to develop new medications for the treatment of central nervous system-related diseases as AD by acting as dual inhibitors of AChE and BChE.

Methyl benzoate derivatives as inhibitors of pentose phosphate pathway, which promotes cancer progression and drug resistance: An in silico study supported by in vitro results.

The pentose phosphate pathway (PPP), whose products are vital in biosynthetic events, is targeted in the treatment of many diseases such as cancer and malaria. The objective of this study was to identify new PPP inhibitors. The inhibition effects of methyl 4-amino benzoates on glucose 6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) were analyzed through in vitro experiments and molecular docking studies were used to estimate inhibition mechanisms. IC50 values of compounds were found between 100.8 and 430.8 µM for G6PD and 206 and 693.2 µM for 6PGD. Molecular docking analysis showed that compound 1 was found the most effective inhibitor against hG6PD and compound 4 had the highest inhibitory potency against h6PGD with the estimated binding energy of -6.71 and -7.61 kcal/mol, respectively. In conclusion, it was determined that in vitro and in silico outcomes of the study were highly correlated with each other. The structure of these benzoates may aid in the development of drugs that target the PPP.

Antibacterial properties and carbonic anhydrase inhibition profiles of azido sulfonyl carbamate derivatives.

Aim: The aim of this study was to identify inhibition of carbonic anhydrase I and II (CA I and II) isozymes by azido sulfonyl carbamates through both in vitro and in silico approaches and also to determine the drug-likeness properties and antibacterial activities of azido sulfonyl carbamates. Methods & Results:In vitro inhibition and molecular docking studies of azido sulfonyl carbamate derivatives (1-4) on isozymes were performed. Except for derivative 4, all derivatives inhibited human CA I and II. Almost all compounds had antibacterial effects. The docking results showed that compound 3 had the best results, with binding energy of -8.20 kcal/mol for human CA I and -8.24 kcal/mol for human CA II. Conclusion: Molecule 4 inhibited only CA I. Its usage as a potential chemotherapeutic agent specific to the CA I isozyme may be considered.

A study on the effects of inhibition mechanism of curcumin, quercetin, and resveratrol on human glutathione reductase through in vitro and in silico approaches.

Glutathione reductase (GR) is a major antioxidant enzyme essential to maintain GSH/GSSG ratio by catalyzing recovery of reduced glutathione (GSH) from oxidized glutathione (GSSG). Because of this vital task, the inhibition of GR is an important target in the treatment of many diseases, so we aimed to identify natural and new GR inhibitors to be guide for drug design.For this purpose, two different approaches were used. The first one is in vitro inhibition, the first phase of which was the purification of the enzyme from human erythrocyte by 2', 5'-ADP Sepharose 4B affinity chromatography, and then the in vitro inhibition effects of curcumin, quercetin, and resveratrol were examined. The second one is in silico study, which was performed to elucidate the drug-likeness, active site identification and inhibition mechanisms of these compounds.hGR was isolated from human erythrocytes with 7.036 EU/mg protein specific activity and 48.97% yield. Then, IC50 values were as 17.25 ± 3.8 µM, 57.8 ± 14.2 µM, and 520 ± 96.7 µM for curcumin, quercetin, and resveratrol respectively. Docking studies of compounds were performed against hGR receptors with induced-fit docking method. The compound showed Glide score as 10.519 kcal/mol, -9.789, and -8.133 respectively.In conclusion, it was seen that curcumin is the much better inhibitor than quercetin and resveratrol for hGR according to both in vitro and in silico studies. Curcumin, a potential inhibitor of hGR, can be used in drug design to target the glutathione system in cellular injury.Communicated by Ramaswamy H. Sarma.

Radical Scavenging and Antiacetylcholinesterase Activities of Ethanolic Extracts of Carob, Clove, and Linden.

CONTEXT: Acetylcholine (ACh) breaks down in a very short time in diseases related to memory loss. It's a neurotransmitter involved in cholinergic transmission in the brain. Acetylcholinesterase (AChE) hydrolyzes ACh. When AChE is inhibited, the ACh levels increase in the cholinergic synapses. The investigation of natural AChE inhibitors with minimal side effects has become important. CONTEXT: Objective • This study intended to determine the total phenolic content, total flavonoid contents, radical scavenging activities, and antiacetylcholinesterase activities of ethanolic extracts of carob pods (ceratonia siliqua), clove buds (eugenia aromatica), and linden flowers (tilia cordata). CONTEXT: Design • The research team designed an in-vitro study. CONTEXT: Setting • The study took place at a biochemistry research laboratory where purification of enzymes and studies on their kinetic properties and inhibitions are carried out. CONTEXT: Outcome measures • The antioxidant properties of the extracts including the total phenolic content (TPC), total flavonoid content (TFC), and free radical scavenging activities, were determined. The AChE enzyme was partially purified by DE-52 anion exchange chromatography from human erythrocytes. Besides, The AChE inhibitory properties of the ethanolic extracts were investigated. CONTEXT: Results • The TPCs of the carob pods, clove buds, and linden flowers were 46.78 ± 0.020, 103.57 ± 0.020, and 28.81 ± 0.031, mg GAE/L, respectively. The TFCs were 27.35 ± 0.021, 30.85 ± 0.017, and 32.12 ± 0.022 mg QE/L, respectively. While the extracts of carob pods and linden flowers inhibited AChE, with IC50s of 0.838 mg/ml and 0.156 mg/ml, respectively, clove buds didn't show inhibitory effect. CONTEXT: Conclusion • Although the clove buds had the maximum TPC; 1,1-diphenyl-2-picryl hydrazyl (DPPH); and 2,2'-azino-bis [3-ethylbenzothiazoline-6-sulphonic acid] (ABTS+) radical scavenging activity, it didn't show anticholinesterase activity.

An in vivo and in vitro comparison of the effects of amoxicillin, gentamicin, and cefazolin sodium antibiotics on the mouse hepatic and renal glutathione reductase enzyme.

Despite the fact that the use of antibiotics is increasing worldwide, it is clear that antibiotics can lead to oxidative stress. This is the first study to make a comparison of the impact of frequently prescribed antibiotics, including amoxicillin, gentamicin, and cefazolin sodium, on the gene, protein, and activity of glutathione reductase (GR), which is one of the primary antioxidant enzymes, in mouse liver and kidney tissues. First, the GR enzyme was purified by the 2',5'-ADP Sepharose 4B affinity chromatography with a specific activity of 84.615 EU/mg protein and 9.63 EU/mg protein from the mouse liver and kidney, respectively. The in vitro inhibitory effects of the antibiotics in question was determined. While cefazolin sodium did not exhibit any inhibitory effect, gentamicin and amoxicillin inhibited GR activity in both tissues. Furthermore, the in vivo effects of these drugs were investigated, and amoxicillin and cefazolin sodium-inhibited GR activity in both liver and kidney tissues, while gentamicin did not have any effect on the kidney. Besides, while gentamicin downregulated and cefazolin sodium upregulated Gr gene expression, amoxicillin did not alter it. Protein expression was only affected by the administration of cefazolin sodium in the kidney. This study is important as it demonstrates that while amoxicillin and gentamicin showed parallel effects on the GR activity in liver and kidney tissues both in vitro and in vivo, cefazolin sodium had a very strong effect on hepatic and renal GR in vivo. Furthermore, the antibiotics used in this study induced oxidative stress in both tissues.