Methylenedioxy Piperamide-Derived Compound D5 Regulates Inflammatory Cytokine Secretion in a Culture of Human Glial Cells.

Neuroinflammation is the cornerstone of most neuronal disorders, particularly neurodegenerative diseases. During the inflammatory process, various pro-inflammatory cytokines, chemokines, and enzymes-such as interleukin 1-ß (IL1-ß), tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), inducible nitric oxide synthases (iNOS), inhibitory kappa kinase (IKK), and inducible nitric oxide (NO)-are over-expressed in response to every stimulus. Methods: In the present study, we focused on the anti-neuroinflammatory efficacy of (2E,4E)-N,5-bis(benzo[d][1,3]dioxol-5-yl)penta-2,4-dienamide, encoded D5. We investigated the efficacy of D5 on the upstream and downstream products of inflammatory pathways in CHME3 and SVG cell lines corresponding to human microglia and astrocytes, respectively, using various in silico, in vitro, and in situ techniques. Results: The results showed that D5 significantly reduced the level of pro-inflammatory cytokines by up-regulating PPAR-γ expression and suppressing IKK-ß, iNOS, NO production, and NF-κB activation in inflamed astrocytes (SVG) and microglia (CHME3) after 24 h of incubation. The data demonstrated remarkably higher efficacy of D5 compared to ASA (Aspirin) in reducing NF-κB-dependent neuroinflammation. Conclusions: We observed that the functional-group alteration had an extreme influence on the levels of druggability and the immunomodulatory properties of two analogs of piperamide, D5, and D4 ((2E,4E)-5-(benzo[d][1,3]dioxol-5-yl)-N-(4-(hydroxymethyl)phenyl)penta-2,4-dienamide)). The present study suggested D5 as a potential anti-neuroinflammatory agent for further in vitro, in vivo, and clinical investigations.

The inhibitory role of benzo-dioxole-piperamide on the phosphorylation process as an NF-Kappa B silencer.

NF-κB contributes to the biosynthesis of various chemokines, cytokines, and enzymes. It plays many crucial roles in the upstream neuroinflammatory pathways. Briefly, the inhibitory IkB subunit is cleaved and phosphorylated by the IKK-α/ß enzyme. It leads to the activation and translocation of the NF-κB (p50/p65) complex into the nucleus. Subsequently, the activated NF-κB interacts with the genomic DNA and contributes to expressing various proinflammatory cytokines. In the present study, we developed a novel NF-κB inhibitor encoded (D5) and investigated the efficacy of our druggable compound through several in silico, in vitro, and in situ analysis. The results demonstrated that D5 not only inhibited the mRNA expression of the IKK-α/ß enzyme (around 86-96% suppression rate for both cell lines at 12 and 24 h time frames) but also by interacting to the active site of the mentioned kinase (dock score -6.14 and binding energy -23.60 kcal/mol) reduced the level of phosphorylated IkB-α in the cytosol around 96-99% and p65 subunit in the nucleus around 73-90% (among all groups in 12 and 24 h time points). Additionally, the results indicated that D5 suppressed the NF-κB target mRNA levels of TNF-α and IL-6 in a total average of around 92%. Overall, The results demonstrated that D5 in a considerably lower concentration than Dis (0.71 µM vs. 52.73 µM) showed significantly higher inhibitory efficacy on NF-κB translocation approx. 200-300%. The results suggested D5 as a potent NF-κB silencer, but further investigations are required to validate our outcomes.

The critical role of piperamide derivative D4 in the regulation of inflammatory response by the microglia and astrocytic glial cells.

Acute and chronic inflammation in the central nervous system plays a critical role in the development of neurodegenerative disorders. Various pro-inflammatory cytokines, chemokines, and enzymes such as TNF-α, IL1-ß, IL-6, COX-1, COX-2, iNOS, IKK, and inducible nitric oxide are expressed in several signalling pathways, and mediate the neuroinflammatory process. ROS and NF-kB nuclear translocation are the two fundamental pathways involved in neuroinflammatory pathogenesis in neuronal and glial cells. In recent years several compoundswere designed to affect the neuroinflammation and suppress neurodegenerative process. Derivatives of natural products (NPs) attract the most attention of drug developers and industries due to their safety and lesser side effects in comparison with generic drugs. One of the most well-known NP is piperine, which is a yellow crystalline alkaloid extracted from black and white pepper. Recently, we developed a novel piperine derivative (((2E,4E)-5-(benzo[d][1,3]dioxol-5-yl)-N-(4-(hydroxymethyl)phenyl)penta-2,4-dienamide, D4) to enhance the specificity and efficacy of the base molecule. Next, we evaluated the potential anti-inflammatory properities of D4 in CHME3 and SVG cell-lines corresponding to human microglia and astrocytes, respectively. Our results indicated that D4 inhibited NF-kB translocation pathway, and significantly reduced transcript and protein levels of pro-inflammatory cytokines in comparison with Aspirin, as a well-known non-selective NSAID. Furthermore, in silico study showed excellent D4 bioavailability in oral administration. The results of the present study suggest a novel molecule with high anti-neuroinflammatory potency for further pre-clinical tests and pharmacological drug investigation.

Impact of novel N-aryl substituted piperamide on NF-kappa B translocation as a potent anti-neuroinflammatory agent.

NF-kB translocation is the key point in the upstream neuroinflammatory pathways. It plays an import role in the pro-inflammatory chemokine, cytokine, and various enzyme expressions, consequently leading to the inflammatory response of the innate immune system. The NF-kB complex consists of structural homolog subunits, including c-Rel, RelB, p52, p65, and p50. Among the p65 subunit has a vital function of NF-kB translocation and DNA binding. NF-kB translocation may occur due to acetylation and phosphorylation LYS 310 and SER311 amino acids in chain A of the p65 subunit in response to IKK-α/ß activity. Therefore, there are two ways to inhibit the NF-kB translocation, either directly blocking the active sites of IKK-α/ß enzymes or protecting the LYS 310 and SER311 of p65 subunit from acetylation and phosphorylation. NF-kB translocation inhibitors can maintain the NF-kB complex in the inactive form inside the cytosol. In this study, we have designed and developed an NF-kB translocation inhibitor, D4. We have performed various in silico, in vitro and in situ studies on the anti-neuroinflammatory function of D4. It showed the ability to inhibit IKK-α/ß in both genome and proteome levels and protect LYS310 of the p65 subunit of NF-kB from the acetylation process. Therefore, we can suggest D4 as the promising anti-neuroinflammatory agent with a function on the upstream process of inflammatory pathways.

Rational Druggability Investigation Toward Selection of Lead Molecules: Impact of the Commonly Used Spices on Inflammatory Diseases.

Herbal remedies and phytochemicals have been used in traditional medicine. Most of the herbs used in human diet have some major effective elements that can affect various pathways in the human body and play a therapeutic role in healing disorders or diseases. Among the inflammatory diseases, worldwide common disorders possess well-known pathways that can be controlled by diet and behavior. There are some well-established targets that are used for anti-inflammatory drugs like cyclooxygenase type 1 and 2 (COX-1 and COX-2), lipoxygenase, prostaglandin D2 receptor, DP1, CRTH2, and so on. In this article, we investigated the role of phytochemicals, extracted from different commonly used spices in the food industry, in preventing or healing the inflammatory disorders. The ability of such bioactives to inhibit COX-2 enzyme has been investigated and compared with marketed selective and nonselective NSAIDs, aspirin and celecoxib. Thereafter, the pharmacokinetic and pharmacodynamic properties of such ingredients have been evaluated for their druggability potential. The results indicated that piperine showed the best ADME (absorption, distribution, metabolism, and excretion) and toxicity profiles among all bioactives. Also, it possessed better affinity value, -7.80518 kcal/mol and energy binding -85.08 kcal/mol, in inhibition of COX-2 with PDB Id: 1CVU rather than other compounds and significantly the higher dock score than aspirin, close to celecoxib. Therefore, piperine has been suggested to be used as the major ingredient in daily diet as a potent anti-inflammatory and anticancer agent.

Impact of novel N-aryl piperamide NO donors on NF-κB translocation in neuroinflammation: rational drug-designing synthesis and biological evaluation.

NO donor drugs showed a significant therapeutic effect in the treatment of many diseases, such as arteriopathies, various acute and chronic inflammatory conditions, and several degenerative diseases. NO-releasing anti-inflammatory drugs are the prototypes of a novel class of compounds, combining the pharmacological activities of anti-inflammatory and anti-nociceptive of drugs with those of NO, thus possessing potential therapeutic applications in a great variety of diseases. In this study, we designed and predicted biological activity by targeting cyclooxygenase type 2 (COX-2) and NF-κB subunits and pharmacological profiling along with toxicity predictions of various N-aryl piperamides linked via an ester bond to a spacer that is bound to a NO-releasing moiety (-ONO2). The result of absorption, distribution, metabolism and excretion and Docking studies indicated that among 51 designed molecules PA-3'K showed the best binding potential in both the substrate and inhibitory binding pocket of the COX-2 enzyme with affinity values of -9.33 and -5.12 for PDB ID 1CVU and 3LN1, respectively, thereby having the potential to be developed as a therapeutic agent. The results of cell viabilities indicated that PA-3'k possesses the best cell viability property with respect to its dose (17.33 ng/ml), with 67.76% and 67.93% viable cells for CHME3 and SVG cell lines, respectively.

Risk of Late-Onset Alzheimer's Disease by Plasma Cholesterol: Rational In Silico Drug Investigation of Pyrrole-Based HMG-CoA Reductase Inhibitors.

Alzheimer's disease (AD), a worldwide renowned progressive neurodegenerative disorder, is the most common cause of dementia. There are several studies on the important role of cholesterol metabolism in AD pathogenesis, which indicated that the high concentrations of serum cholesterol increase the risk of AD. Biosynthesis of the plasma cholesterol and other isoprenoids is catalyzed by 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) through the conversion of HMG-CoA to mevalonic acid in mevalonate pathway. Normally, the high level of plasma cholesterol is downregulated by HGMCR inhibition as the result of degradation of LDL, but in abnormal conditions, for example, high blood glucose, the HMGCR over activated resulting in uncontrolled blood cholesterol. Selective HMGCR inhibitor drugs such as statins, which increase the catabolism of plasma LDL and reduce the plasma concentration of cholesterol, have been investigated as a possible treatment for AD. In the present study, we have identified the binding modes of 22 various derivatives of 3-sulfamoylpyrroles 16, prepared via a [3 + 2] cycloaddition of a münchnone with a sulfonamide-substituted alkyne, by using efficient biocomputational tools. Out of 22, 5 ligands, with code numbers 5b, 5c, 5d, 5i, and 5j, possessed most absorption, distribution, metabolism, and excretion (ADME) and toxicity profiles in acceptable ranges. Among ligands, 5j (sodium (3R,5R)-7-(3-(N,N-dimethylsulfamoyl)-5-(4-fluorophenyl)-2-isopropyl-4-phenyl-1H-pyrrol-1-yl)-3,5-dihydroxyheptanoate) could inhibit HMGCR enzyme in inhibitory binding site with affinity value -12.17 kcal/mol and binding energy -94.10 kcal/mol through 5 hydrogen bonds. It showed the best ADME and toxicity profiling and higher affinity values than other potent candidate and market drugs such as atorvastatin and rosuvastatin. Therefore, it is suggested for further in vivo investigation, the druggability of 5j and its cholesterol regulatory impact on AD.

Chemometric profile & antimicrobial activities of leaf extract of Calotropis procera and Calotropis gigantea.

Calotropis procera and Calotropis gigantea are medicinal plant having therapeutic value. The leaf extracts of C. procera have been investigated, its pharmacological actions in detail and leaf extracts of C. gigantea were not studied till date. The objective of present work was to find the bioactive constituents present in the ethanolic leaf extract of C. procera and C. gigantea to evaluate their antibacterial and anifungal activities. The major phytochemical groups in C. procera ethanolic leaf extracts were fatty acid ethyl ester (21.36%), palmitic acid ester (10.24%), linoleic acid (7.43%) and amino acid (8.10%) respectively, whereas ethanolic leaf extracts of C. gigantea contain palmitic acid (46.01%), diterpene (26.53%), triterpene (17.39%), linoleic acid (5.13%) as the major phytochemical groups. Ethanol extract of C. procera leaves showed the highest inhibition (11 mm) against Escherichia coli, while ethanolic extract of C. gigantea leaves inhibited Klebsiella (20 mm). These findings will use in new directions in pharmacological investigations.

Drug Targets for Cardiovascular-Safe Anti-Inflammatory: In Silico Rational Drug Studies.

Cyclooxygenase-2 (COX-2) plays an important role in memory consolidation and synaptic activity, the most fundamental functions of the brain. It converts arachidonic acid to prostaglandin endoperoxide H2. In contrast, if over-expressed, it causes inflammation in response to cytokine, pro-inflammatory molecule, and growth factor. Anti-inflammatory agents, by allosteric or competitive inhibition of COX-2, alleviate the symptoms of inflammation. Coxib family drugs, particularly celecoxib, are the most famous anti-inflammatory agents available in the market showing significant inhibitory effect on COX-2 activity. Due to high cardiovascular risk of this drug group, recent researches are focused on the investigation of new safer drugs for anti-inflammatory diseases. Natural compounds, particularly, phytochemicals are found to be good candidates for drug designing and discovery. In the present study, we performed in silico studies to quantitatively scrutinize the molecular interaction of curcumin and its structural analogs with COX-2, COX-1, FXa and integrin αIIbßIII to investigate their therapeutic potential as a cardiovascular-safe anti-inflammatory medicine (CVSAIM). The results of both ADMET and docking study indicated that out of all the 39 compounds studied, caffeic acid had remarkable interaction with proteins involved in inflammatory response. It was also found to inhibit the proteins that are involved in thrombosis, thereby, having the potential to be developed as therapeutic agent.

Molecular modeling, docking and ADMET studies towards development of novel Disopyramide analogs for potential inhibition of human voltage gated sodium channel proteins.

The sodium "channelopathies" are the first among the ion channel diseases identified and have attracted widespread clinical and scientific interests. Human voltage gated sodium channels are sites of action of several antiarrhythmic drugs, local anesthetics and related antiepileptic drugs. The present study aims to optimize the activity of Disopyramide, by modification in its structures which may improve the drug action by reducing its side effects. Herein, we have selected Human voltage-gated sodium channel protein type 5 as a potent molecular target. Nearly eighty analogs of Disopyramide are designed and optimized. Thirty are selected for energy minimization using Discovery studio and the LigPrep 2.5. Prior to docking, the active sites of all the proteins are identified. The processing, optimization and minimization of all the proteins is done in Protein preparation wizard. The docking study is performed using the GLIDE. Finally top five ranked lead molecules with better dock scores are identified as having strong binding affinity to 2KAV protein than Disopyramide based on XP G scores. These five leads are further docked with other similar voltage gated sodium channel proteins (PDB IDs: 2KBI, 4DCK, 2L53 and 4DJC) and the best scoring analog with each protein is identified. Drug likeliness and comparative bioactivity analysis for all the analogs is done using QikProp 3.4. Results have shown that the top five lead molecules would have the potential to act as better drugs as compared to Disopyramide and would be of interest as promising starting point for designing compounds against various Sodium channelopathies.