Cinnamic acids as promising bioactive compounds for cancer therapy by targeting MAPK3: a computational simulation study.

Matrix metalloproteinase-9 (MMP-9) is a zinc and calcium-dependent proteolytic enzyme involved in extracellular matrix degradation. Overexpression of MMP-9 has been confirmed in several disorders, including cancers, Alzheimer's disease, autoimmune diseases, cardiovascular diseases, and dental caries. Therefore, MMP-9 inhibition is recommended as a therapeutic strategy for combating various diseases. Cinnamic acid derivatives have shown therapeutic effects in different cancers, Alzheimer's disease, cardiovascular diseases, and dental caries. A computational drug discovery approach was performed to evaluate the binding affinity of selected cinnamic acid derivatives to the MMP-9 active site. The stability of docked poses for top-ranked compounds was also examined. Twelve herbal cinnamic acid derivatives were tested for possible MMP-9 inhibition using the AutoDock 4.0 tool. The stability of the docked poses for the most potent MMP-9 inhibitors was assessed by molecular dynamics (MD) in 10 nanosecond simulations. Interactions between the best MMP-9 inhibitors in this study and residues incorporated in the MMP-9 active site were studied before and after MD simulations. Cynarin, chlorogenic acid, and rosmarinic acid revealed a considerable binding affinity to the MMP-9 catalytic domain (ΔGbinding < -10 kcal/ mol). The inhibition constant value for cynarin and chlorogenic acid were calculated at the picomolar scale and assigned as the most potent MMP-9 inhibitor from the cinnamic acid derivatives. The root-mean-square deviations for cynarin and chlorogenic acid were below 2 Å in the 10 ns simulation. Cynarin, chlorogenic acid, and rosmarinic acid might be considered drug candidates for MMP-9 inhibition.

Chlorogenic Acids from Green Coffee Extract are Highly Bioavailable in Humans

Capparis spinosa L. (caper) is an important medicinal plant whose bioactive compounds vary significantly depending on its growth stage. This directly affects its pharmaceutical and nutritional value. Collecting C. spinosa at the optimal growth stage is essential to achieving high phytochemical quality and meeting consumer needs. This study aimed to evaluate the variation of these active compounds in the aerial parts of C. spinosa across four phenological stages (vegetative, flowering, unripe fruit, and ripe fruit). The result showed that EO content was highest in unripe fruits (0.18%) and lowest in the flowering stage (0.07%) in leaves, while extract yield was highest in leaves of the ripe fruit stage (14.65%) followed by the flowering stage in flowers (12.66%). Flowering stage leaves showed the highest total phenol (56.20 mg GAE/g) and flavonoid (17.10 mg QE/g) content, while the lowest concentrations were found in the ripe fruit stage of the leaves. EO analysis showed that methyl isothiocyanate reached the highest concentration in flowers at the flowering stage (41.6%), while isopropyl isothiocyanate reached the highest concentration in leaves at the ripe fruit stage (36.2%). Isobutyl isothiocyanate was found exclusively in fruits, with the highest concentration in ripe fruits (9.2%). Dimethyltrisulphide showed a maximum concentration in leaves at the vegetative stage, decreasing by 76.6% as the plant developed towards the ripe fruit stage. The dominant phenolic acids varied between phenological stages: cinnamic acid at the vegetative stage; rosmarinic and cinnamic acids at the flowering stage in leaves; caffeic and cinnamic acids in flowers; vanillic, cinnamic, and rosmarinic acids at the unripe fruit stage in leaves and fruits; and rosmarinic, cinnamic, and vanillic acids in ripe fruits. The results indicate that harvesting C. spinosa at the vegetative stage and in the leaves of the flowering stage is optimal for maximum secondary metabolite yield, providing valuable guidance for growers targeting food and pharmaceutical applications.

Five wild edible mushrooms (Amanita caesarea, Boletus edulis, Cantharellus cibarius, Lactarius indigo and Ramaria sp.) from Mexico were studied for their total phenolic content, antioxidant activity, flavonoids, ascorbic, gallic, cinnamic and chlorogenic acids. B. edulis showed the highest contents expressed in dry weight, of flavonoids (0.92 mg of quercetin equivalents/g), total phenolic acids (8.66 mg of gallic acid equivalents/g), chlorogenic acid (1001.67 ?g/g) and cinnamic acid (10.68 ?g/g) it also presented the greatest antioxidant activity (48.6% DPPH scavenging activity and 59.48% reductive power) from the species analysed. Overall, Mexican edible mushrooms had high concentrations of chlorogenic and ascorbic acids, and flavonoids. This study constitutes the first report in Mexico about nutraceuticals from wild edible mushrooms.

Phenolic acids and phenylpropanoids have an important biological activity and are therapeutic agents of crude drugs. Development of validated analysis techniques of these phytotherapeutic agents (fingerprinting and assay procedures) is an important practice for efficacy, safety, and quality control of herbal drug preparations. The aim of the present work was to study analytical capabilities of the evaluation of selected phenolic acids and phenylpropanoids: caffeic acid, chlorogenic acid, cinnamic acid, coumaric acid, ferulic acid, gallic acid, protocatechuic (3,4-dihydroxybenzoic) acid, rosmarinic acid, vanillic acid, and vanillin. Optimization and validation procedures of rapid and simple method of reversed-phase high-performance liquid chromatography were carried out. The mobile phase of the optimized chromatographic method consisted of methanol and 0.5% acetic acid solvent in water. For the application of method, two kinds of raw materials were chosen: propolis and the Herba Origani. Coumaric acid is the dominating phenolic acid of propolis (2785 microg/g). Results of analysis of Herba Origani demonstrated high quantities (6376 microg/g) of rosmarinic and protocatechuic (1485 microg/g) acids in the samples.

Background: A common oral inflammatory disease known as apical periodontitis (AP) is caused by the intrusion of microorganisms into the dental pulp, resulting in an inflammatory response and bone degradation in periapical tissues. A growing body of evidence indicates that the receptor-interacting serine-threonine kinase 3 (RIPK3) is closely associated with AP. Objectives: This study sought to address the requirement for effective RIPK3 inhibitors by examining the potential of cinnamic acid natural metabolites capable of inhibiting RIPK3. Methods: The binding affinity of 20 cinnamic acids to the RIPK3 active site was evaluated by using AutoDock 4.0 software. The most favorable scores were assigned to the highest-ranking cinnamic acids based on their ΔGbinding values to the RIPK3 catalytic domain. A 100-nanosecond (ns) computer simulation was performed using molecular dynamics for the most efficacious inhibitor of RIPK3, and the findings were contrasted with those obtained for free RIPK3. The Discovery Studio Visualizer tool was employed to showcase the interactions between the RIPK3 active site and the highest-ranking metabolites. Results: The binding affinity of cynarin, rosmarinic acid (RosA), and chlorogenic acid (CGA) to the RIPK3 active site was noteworthy, as the ΔGbinding values were&lt;-10 kcal/mol. Furthermore, cynarin exhibited inhibition constant values at the picomolar range. Upon complexation with cynarin, the RIPK3 conformation attained stability after approximately 25 ns of simulation. Conclusion: In general, cynarin, RosA, and CGA have the potential to be therapeutically beneficial in treating AP due to their ability to inhibit RIPK3.

Leaf disks of Nicotiana tabacum var. Delcrest were supplied with α-C14-labelled cinnamic, p-coumaric, and caffeic acids, p-coumaroyl-quinic acid, and the 1-glucose esters of cinnamic and p-coumaric acids. Considerable incorporation of radioactivity into chlorogenic acid occurred in 40 hours from all substrates except caffeic acid. In particular, p-coumaric acid was found to be a good precursor of chlorogenic acid in contrast to the findings of other workers with potato tuber tissue. More p-coumaric acid than cinnamic acid is converted to chlorogenic acid, although the reverse holds for conversion to p-coumaroyl-quinic acid. Chlorogenic acid synthesis from caffeic acid involves breakdown and resynthesis of the acid supplied, leading to the incorporation of radioactivity into other positions in the caffeic acid molecule and into the quinic acid moiety. Cinnamic and p-coumaric acids, on the other hand, are incorporated essentially intact into both p-coumaroyl-quinic and chlorogenic acids. These findings are discussed in relation to the pathway of chlorogenic acid biosynthesis postulated for potato tuber tissue.

Objectives: In this study, using HPLC-ED system, analysis of chlorogenic acid (CGA), gallic acid (GA), rosmarinic acid (RA) and caffeic acid (CA), and rutin was carried out in water extracts of sage and different thyme. Methods: Analyses of CGA, GA, RA, CA and rutin were performed of herbs sage and thyme. The drug (1g) was powdered and extracted with pure water (9ml). Afterward 1ml of that extract was decanted and centrifuged. Supernatant was used for analysis. The standard solutions were of CGA, GA, RA, and CA dissolved in mobile phase, and rutin was dissolved pure water. HPLC conditions were following: Mobile phase methanol-acetonitrile-water-acetic acid (20+10+70+1); ED detector with range 50nA, potential +0.840 V, filter 0.02Hz; flow rate 1ml/min; temperature 25°C, Column: ODS hypersil. Results: The content in (mg/g) of CGA was in the sage 0.36, GA 0.09, RA 5.45, CA 0.73, and rutin 11.8, thyme: (Thymus vulgaris L. CGA 1.21, GA 0.3, RA 4.27, CA 0.66, and rutin 24.9, Thymus serpyllum L. CGA 4.6, GA 0.95, RA 17.27, CA 1.36, and rutin 17.27, Thymus sibthorii Benth. CGA 1.44, GA 0.3, RA 5.5, CA 0.95, and rutin 22.64). Conclusion: The highest content of CGA, RA, CA and GA was determined in herbs of Thymus serpyllum L., and highest content of rutin was determined in Thymus vulgaris L.

The ERK1/2 MAP kinase pathway is an evolutionarily conserved signaling module that controls many fundamental physiological processes. Deregulated activity of ERK1/2 MAP kinases is associated with developmental syndromes and several human diseases. Despite the importance of this pathway, a comprehensive picture of the natural substrate repertoire and biochemical mechanisms regulated by ERK1/2 is still lacking. In this study, we used large-scale quantitative phosphoproteomics and bioinformatics analyses to identify novel candidate ERK1/2 substrates based on their phosphorylation signature and kinetic profiles in epithelial cells. We identified a total of 7936 phosphorylation sites within 1861 proteins, of which 155 classify as candidate ERK1/2 substrates, including 128 new targets. Candidate ERK1/2 substrates are involved in diverse cellular processes including transcriptional regulation, chromatin remodeling, RNA splicing, cytoskeleton dynamics, cellular junctions and cell signaling. Detailed characterization of one newly identified substrate, the transcriptional regulator JunB, revealed that ERK1/2 phosphorylate JunB on a serine adjacent to the DNA-binding domain, resulting in increased DNA-binding affinity and transcriptional activity. Our study expands the spectrum of cellular functions controlled by ERK1/2 kinases.

Hydroxycinnamic acid derivatives are naturally occurring substances found in fruits, vegetables, and flowers and are consumed as dietary phenolic compounds. The effect of cinnamic acid, ferulic acid, p-coumaric acid, eugenol, chlorogenic acid, and caffeic acid, alone and in combination with two commercial oral hypoglycemic drugs (OHD), namely, thiazolidinedione (THZ) and metformin, on the uptake of 2-deoxyglucose (2DG) by 3T3-L1 adipocytes is studied. All of the phytochemicals other than cinnamic acid show synergistic interaction in 2DG uptake with both of the OHDs. THZ (20 μM) in combination with ferulic acid (25 μM) or p-coumaric acid (25 μM) increases 2DG uptake by 7- or 6.34-fold, respectively, with respect to control, whereas metformin (20 μM), along with ferulic acid (25 μM) or cinnamic acid (25 μM), increases 2DG uptake by 6.45- or 5.87-fold, respectively, when compared to control. Chlorogenic and cinnamic acids increased the expression of PPARγ, whereas other hydroxycinnamic acids enhanced the expression of PI3K, indicating different mechanisms of action between these compounds. These phytochemicals were able to reduce the expressions of the fatty acid synthase and HMG CoA reductase genes, indicating that they may be able to reduce the secondary complications caused by the accumulation of lipids. These studies suggest that hydroxycinnamic acid derivatives may be beneficial for the treatment of diabetes mellitus. They may act as a supplement with commercial drugs and may reduce the secondary complications caused by OHDs.

L'oxydation des lipides insatures, qui constitue l'une des causes majeures de l'alteration des produits alimentaires et cosmetiques, peut etre limitee par l'emploi d'antioxydants. Les composes phenoliques, et plus particulierement les acides phenoliques et les flavonoides, suscitent un interet grandissant pour leurs proprietes antioxydantes. Cependant, la mise en oeuvre de ces molecules polaires dans les matrices lipidiques (de type emulsion ou autre) est delicate et peut s'accompagner d'une diminution de leur efficacite. Pour contourner ce probleme, l'une des strategies consiste a ajuster la polarite de ces molecules par le greffage de chaines aliphatiques de longueurs differentes. La premiere partie de ce travail a ete consacree a la synthese enzymatique d'une serie d'esters lipophiles des acides chlorogenique et rosmarinique. Concernant l'acide chlorogenique, les meilleurs resultats ont ete obtenus en utilisant une strategie en deux etapes. Cette strategie correspond a une esterification prealable au methanol en presence d'une resine sulfonique suivie d'une transesterification enzymatique (lipase B de Candida antarctica) du chlorogenate de methyle en milieu fondu, avec des alcools de longueurs de chaine comprise entre 4 et 16 atomes de carbone. En conditions optimales, les vitesses initiales de transesterification s'averent jusqu'a deux fois superieures aux vitesses d'esterification enzymatique directe. Par ailleurs, la strategie en deux etapes conduit a des rendements globaux de 61 a 93% contre 40 a 60% dans le cas de l'esterification directe. Concernant l'acide rosmarinique, seule l'esterification directe en presence de resine sulfonique a permis d'atteindre des rendements satisfaisants (de 82% a 99%, selon la longueur de la chaine greffee). Dans la deuxieme partie de ce travail, nous avons evalue, d'un point de vue cinetique et stationnaire, l'impact de la lipophilisation sur la capacite des nouvelles molecules a pieger le radical stable 2,2-diphenyl-1-picryihydrazyl (DPPH*). Les donnees cinetiques montrent que l'acide chlorogenique se stabilise sous sa forme quinone, tandis que les esters tendent a former des dimeres a partir de leurs formes semi-quinone. Les resultats obtenus a l'etat stationnaire indiquent que les acides chlorogenique et rosmarinique, mais egalement leurs esters . respectifs, ont une capacite reductrice superieure a la capacite theorique imputable aux nombre d'hydroxyles phenoliques. Ces resultats peuvent etre expliques en considerant l'existence de reactions de regeneration et/ou de dimerisation. Enfin, la troisieme partie de ce travail a ete consacree a l'evaluation de la capacite antioxydante des derives lipophilises d'acides chlorogenique et rosmarinique en milieu emulsionne, par une nouvelle methodologie appelee test des Menes conjugues autoxydables (CAT). Les resultats ont montre que l'augmentation de la longueur de chaine aliphatique ne conduisait pas forcement a une amelioration de la capacite antioxydante comme le predit le «Paradoxe Polaire». En effet, un maximum de la capacite antioxydante pour des longueurs de chaine de 12 et 8 atomes de carbone a ete observe pour les derives lipophiles des acides chlorogenique et rosmarinique respectivement. Les principales hypotheses avancees pour expliquer ce comportement inattendu sont basees sur la repartition des antioxydants et de leurs formes oxydees entre les diverses phases du systeme reactionnel. (Resume d'auteur)

Disruption of fungal cell wall should be an effective intervention strategy. However, the cell wall-disrupting echinocandin drugs, such as caspofungin (CAS), cannot exterminate filamentous fungal pathogens during treatment. For potency improvement of cell wall-disrupting agents (CAS, octyl gallate (OG)), antifungal efficacy of thirty-three cinnamic acid derivatives was investigated against Saccharomyces cerevisiaeslt2Δ, bck1Δ, mutants of the mitogen-activated protein kinase (MAPK), and MAPK kinase kinase, respectively, in cell wall integrity system, and glr1Δ, mutant of CAS-responsive glutathione reductase. Cell wall mutants were highly susceptible to four cinnamic acids (4-chloro-α-methyl-, 4-methoxy-, 4-methyl-, 3-methylcinnamic acids), where 4-chloro-α-methyl- and 4-methylcinnamic acids possessed the highest activity. Structure-activity relationship revealed that 4-methylcinnamic acid, the deoxygenated structure of 4-methoxycinnamic acid, overcame tolerance of glr1Δ to 4-methoxycinnamic acid, indicating the significance of para substitution of methyl moiety for effective fungal control. The potential of compounds as chemosensitizers (intervention catalysts) to cell wall disruptants (viz., 4-chloro-α-methyl- or 4-methylcinnamic acids + CAS or OG) was assessed according to Clinical Laboratory Standards Institute M38-A. Synergistic chemosensitization greatly lowers minimum inhibitory concentrations of the co-administered drug/agents. 4-Chloro-α-methylcinnamic acid further overcame fludioxonil tolerance of Aspergillus fumigatus antioxidant MAPK mutants (sakAΔ, mpkCΔ). Collectively, 4-chloro-α-methyl- and 4-methylcinnamic acids possess chemosensitizing capability to augment antifungal efficacy of conventional drug/agents, thus could be developed as target-based (i.e., cell wall disruption) intervention catalysts.

COVID-19-associated mucormycosis (CAM) has emerged as a concerning complication during the COVID-19 pandemic. In this study, we explored the potential of phytochemicals and flavonoids identified through High-Performance Liquid Chromatography (HPLC) analysis of Thymus vulgaris plant extract against key proteins of CAM, namely heat shock protein A5 (GPR78) and epidermal growth factor receptor (EGFR). HPLC analysis revealed the presence of bioactive compounds, including chlorogenic acid, cinamic acid, quercetin, coumaric acid, gallic acid, and syringic acid. To assess their efficacy against CAM, computational analyses were performed, including molecular docking analysis, pharmacophore characterization, ADME and molecular dynamics simulations. The results demonstrated that chlorogenic acid exhibited strong binding affinity against EGFR with a docking score of -7.6 kcal/mol, while quercetin showed favorable binding affinity against HSP A5 (GPR78) with a docking score of -10.1 kcal/mol. Both chlorogenic acid and quercetin displayed promising ADME properties, indicating their potential as drug candidates. Nevertheless, it was observed that chlorogenic acid did not adhere to Lipinski’s rule, and its gastrointestinal (GI) absorption was relatively low when compared to quercetin. Unlike chlorogenic acid, quercetin does conform to Lipinski’s rule and showed high GI absorption. Moreover, pharmacophore characterization of both drug candidates revealed a substantial number of binding sites, suggesting the likelihood of stable bond formation. Normal mode analysis revealed higher eigenvalues for the quercetin–HSPA5 complex compared to the chlorogenic acid–EGFR complex, indicating greater structural rigidity and stability. Overall, our findings highlight the potential of chlorogenic acid and quercetin as promising drug candidates against CAM. Furthermore, in-vitro and in-vivo studies are needed to validate their efficacy and safety for clinical use in treating mucormycosis associated with COVID-19. These findings may offer valuable insights into the development of novel therapeutic options to combat this challenging co-infectious disease.

Lepidium sativum L. (Brassicaceae), garden cress or pepper grass, is the source of valuable biologically active compounds [1]. It is used in folk medicine for cancer, uterine tumors, polyps, and other neoplasms. It is used in medicine as the alcoholic tincture, which possesses sedative and anticonvulsive activity [2]. Herein we communicate results from investigations of phenolic compounds isolated from L. sativum seeds grown in culture by us. Ground air-dried raw material (seeds, 1.0 g) was extracted successively with hexane, CCl4, and CHCl3 (20 mL each), discarding the produced extracts. Then, the remaining raw material was extracted with boiling ethanol (70%) for 1 h. The extract was cooled, filtered through paper into a volumetric flask (100-mL), and adjusted to the mark with ethanol (70%). The resulting extract (2 mL) was placed into a volumetric flask (25-mL) and adjusted with the same solvent to the mark (stock solution). A series of reference solutions in ethanol (70%) was prepared in parallel. These included rutin, quercetin, luteolin, luteolin-7-glycoside, hyperoside, hesperidin, apigenin, vicenin, vitexin, 4-hydroxycoumarin, scopoletin, robinin, dicoumarin, coumarin, umbelliferone, dihydroquercetin, catechin, orientin, and gallic, chlorogenic, neochlorogenic, cinnamic, caffeic, ferulic, salicylic, and ellagic acids. The stock and reference solutions (20 μL) were injected into a chromatograph. The qualitative composition of the phenolic compounds from L. sativum seeds was studied on a Gilston Model 305 (France) high-performance liquid chromatograph (HPLC) with a Rheodyne 7125 (USA) manual injector with subsequent computer processing of the results using the program Multichrom for Windows. The mobile phase was CH3OH:H2O:H3PO4 (400:600:5). The analysis was performed at room temperature. The eluent flow rate was 0.5 mL/min; analysis time, 120 min. Detection used a Gilston Model 151 UV/Vis UV detector operating at 254 nm. A total of 12 compounds was isolated by the HPLC analysis of L. sativum seeds. Of these, gallic acid (9.44%, content in the isolated mixture by internal normalization method), chlorogenic acid (14.77%), ferulic acid (5.63%), neochlorogenic acid (2.22%), luteolin-7-glycoside (14.67%), dihydroquercetin (4.37%), and quercetin (3.15%) were identified. Quantitative analysis of phenolic compounds from L. sativum seeds was based on the UV absorption spectra of this group of compounds. The absorption spectra were defined using the extract (70% ethanol) of seeds and a Helios (USA) selfrecording spectrophotometer in the wavelength range 210-400 nm. According to preliminary results on the identification by HPLC of phenolic compounds from L. sativum seeds, chlorogenic acid was the predominant component. Therefore, the absorption spectrum of chlorogenic acid in ethanol (70%) was recorded in parallel as a working standard in the same wavelength range. Absorption spectra of the studied solution and the chlorogenic acid solution had a common absorption maximum at 329 ± 2 nm. This enabled us to calculate the total phenolic compounds in L. sativum seeds using chlorogenic acid as a working standard. The studied solution was prepared by the method given above (weight 1.0 g). The working standard (chlorogenic acid, 0.02 g) was placed in a volumetric flask (50-mL), treated with ethanol (70%, 30 mL), stirred until dissolved, and adjusted to the mark (solution A). Solution A (1 mL) was placed in a volumetric flask (50-mL) and adjusted with the same solvent to the mark (working standard). The optical density of the resulting solutions was measured at 329 nm relative to ethanol (70%). The contents of total phenolic compounds calculated as chlorogenic acid in L. sativum seeds was 0.85%.

Chlorogenic acid, the ester of caffeic acid with quinic acid, is one of the most abundant polyphenols in the human diet. The antioxidant and anticarcinogenic properties of chlorogenic acid have been established in animal studies. However, little is known about the molecular mechanisms through which chlorogenic acid inhibits carcinogenesis. In this study, we found that chlorogenic acid inhibited the proliferation of A549 human cancer cells in vitro. The results of the soft agar assay indicated that chlorogenic acid suppressed 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced neoplastic transformation of JB6 P+ cells in a dose-dependent manner. Pretreatment of JB6 cells with chlorogenic acid blocked UVB- or TPA-induced transactivation of AP-1 and NF-κB over the same dose range. At low concentrations, chlorogenic acid decreased the phosphorylation of c-Jun NH2-terminal kinases, p38 kinase, and MAPK kinase 4 induced by UVB/12-O-tetradecanoylphorbol-13-acetate, yet higher doses were required to inhibit extracellular signal-regulated kinases. Chlorogenic acid also increased the enzymatic activities of glutathione S-transferases (GST) and NAD(P)H: quinone oxidoreductase. Further studies indicated that chlorogenic acid could stimulate the nuclear translocation of Nrf2 (NF-E2-related factor) as well as subsequent induction of GSTA1 antioxidant response element (ARE)-mediated GST activity. The phosphatidylinositol 3-kinase pathway might be involved in the activation of Nrf2 translocation. These results provide the first evidence that chlorogenic acid could protect against environmental carcinogen-induced carcinogenesis and suggest that the chemopreventive effects of chlorogenic acid may be through its up-regulation of cellular antioxidant enzymes and suppression of ROS-mediated NF-κB, AP-1, and MAPK activation.