Unraveling the mechanism of carbonic anhydrase IX inhibition by alkaloids from Ruta chalepensis: A synergistic analysis of in vitro and in silico data.

Due to the pivotal role of carbonic anhydrase IX (CA IX) in pathological conditions, there's a pressing need for novel inhibitors to improve patient outcomes and clinical management. Herein, we investigated the inhibitory efficacy of six alkaloids from Ruta chalepensis against CA IX through in vitro inhibition assay and computational modeling. Skimmianine and maculosidine displayed significant inhibitory activity in vitro, with low IC50 values of 105.2 ± 3.2 and 295.7 ± 14.1 nM, respectively. Enzyme kinetics analyses revealed that skimmianine exhibited a mixed inhibition mode, contrasting with the noncompetitive inhibition mechanism observed for the reference drug (acetazolamide), as indicated by intersecting lines in the Lineweaver-Burk plots. The findings of docking calculations revealed that skimmianine and maculosidine exhibited extensive polar interactions with the enzyme. These alkaloids demonstrate substantial binding interactions and occupy identical binding site as acetazolamide, thereby enhancing their efficacy as inhibitors of CA IX. Utilizing a 100 ns molecular dynamics (MD) simulation, the dynamic interactions between isolated alkaloids and CA IX were intensively assessed. Analysis of diverse MD parameters revealed that skimmianine and maculosidine displayed consistent trajectories and notable energy stabilization during their interaction with CA IX. The findings of MM/PBSA analysis depicted the minimum binding free energy for skimmianine and maculosidine. In addition, the Potential Energy Landscape (PEL) analysis revealed distinct and stable conformational states for the CA IX-ligand complexes, with Skimmianine showing the most stable and lowest energy configuration. These computational findings align with experimental results, emphasizing the potential efficacy of skimmianine and maculosidine as inhibitors of CA IX.

Chemical Composition, Antioxidant, and Antibacterial Activity of Ruta chalepensis L. Ethanolic Extract.

Ruta chalepensis L. is a versatile herb used in culinary arts and traditional medicine. The study aimed to determine the chemical composition of an ethanolic extract from R. chalepensis and the total phenolic and flavonoid content. Additionally, the extracts' antimicrobial and antioxidant activities were tested. The disc diffusion method and minimum inhibitory concentration (MIC) were used to test the antibacterial properties on four types of bacteria: Escherichia coli, Proteus penneri, Bacillus cereus, and Staphylococcus aureus. A colorimetric assay was used to evaluate the total phenolic and flavonoid content, and the DPPH method was used to assess the antioxidant activity. The phytochemical constituents were determined using LC-MS/MS. The results indicated that R. chalepensis ethanolic extract had 34 compounds, and the predominant compounds were quercetin (9.2 %), myricetin (8.8 %), and camphene (8.0 %). Moreover, the extract had a good level of polyphenols and flavonoids, as demonstrated by inhibiting free radicals (DPPH) (IC50 was 41.2±0.1). Also, the extract exhibited robust antimicrobial activity against P. penneri and S. aureus with an MIC of 12.5 and 25.0 µg/mL, respectively. In conclusion, the results suggest that the R. chalepensis ethanolic extract has good antioxidant and antibacterial properties that could be utilized to develop new antibacterial agents.

Antifungal Mechanism of Ruta graveolens Essential Oil: A Colombian Traditional Alternative against Anthracnose Caused by Colletotrichum gloeosporioides.

Here, we report for the first time on the mechanisms of action of the essential oil of Ruta graveolens (REO) against the plant pathogen Colletotrichum gloeosporioides. In particular, the presence of REO drastically affected the morphology of hyphae by inducing changes in the cytoplasmic membrane, such as depolarization and changes in the fatty acid profile where straight-chain fatty acids (SCFAs) increased by up to 92.1%. In addition, REO induced changes in fungal metabolism and triggered apoptosis-like responses to cell death, such as DNA fragmentation and the accumulation of reactive oxygen species (ROS). The production of essential enzymes involved in fungal metabolism, such as acid phosphatase, ß-galactosidase, ß-glucosidase, and N-acetyl-ß-glucosaminidase, was significantly reduced in the presence of REO. In addition, C. gloeosporioides activated naphthol-As-BI phosphohydrolase as a mechanism of response to REO stress. The data obtained here have shown that the essential oil of Ruta graveolens has a strong antifungal effect on C. gloeosporioides. Therefore, it has the potential to be used as a surface disinfectant and as a viable replacement for fungicides commonly used to treat anthracnose in the postharvest testing phase.

Encapsulation of Ruta essential oil in chitosan and alginate matrices as an ecological alternative for the control of nematodes.

Controlled release formulations of Ruta essential oil obtained by ionic gelation were developed. The presence of rue essential oil in the alginate and chitosan capsules was evidenced by Fourier transform infrared spectroscopy and differential scanning calorimetry. Release studies revealed that in acidic conditions (pH 4.2), the CHS-REO particles reached a Sw of 240% (w/w) in 30 days and 101% (w/w) for ALG-REO particles, generating a RR of 23.7% for CHS-REO and 20.4% for ALG-REO. On the other hand, at pH 6.8 it favored the Sw for ALG-REO 840% (w/w) and therefore the RR (45.6%) and disfavored the Sw of CHS-REO generating low RR (16.9%). Encapsulated rue essential oil showed equal or superior nematicidal activity against the nematode Melodogyne ssp., compared to free oil and a synthetic nematicide such as Carbofuran, without having a phytotoxic effect on the plant. This study revealed that REO encapsulated in biopolymeric matrices can be used as new nematicide formulations.

Studies on the Accumulation of Secondary Metabolites and Evaluation of Biological Activity of In Vitro Cultures of Ruta montana L. in Temporary Immersion Bioreactors.

The present work focuses on in vitro cultures of Ruta montana L. in temporary immersion PlantformTM bioreactors. The main aim of the study was to evaluate the effects of cultivation time (5 and 6 weeks) and different concentrations (0.1-1.0 mg/L) of plant growth and development regulators (NAA and BAP) on the increase in biomass and the accumulation of secondary metabolites. Consequently, the antioxidant, antibacterial, and antibiofilm potentials of methanol extracts obtained from the in vitro-cultured biomass of R. montana were evaluated. High-performance liquid chromatography analysis was performed to characterize furanocoumarins, furoquinoline alkaloids, phenolic acids, and catechins. The major secondary metabolites in R. montana cultures were coumarins (maximum total content of 1824.3 mg/100 g DM), and the dominant compounds among them were xanthotoxin and bergapten. The maximum content of alkaloids was 561.7 mg/100 g DM. Concerning the antioxidant activity, the extract obtained from the biomass grown on the 0.1/0.1 LS medium variant, with an IC50 0.90 ± 0.03 mg/mL, showed the best chelating ability among the extracts, while the 0.1/0.1 and 0.5/1.0 LS media variants showed the best antibacterial (MIC range 125-500 µg/mL) and antibiofilm activity against resistant Staphylococcus aureus strains.

Headspace Solid Phase Micro-Extraction of Volatile Constituents Produced from Saudi Ruta chalepensis and Molecular Docking Study of Potential Antioxidant Activity.

Ruta chalepensis L., commonly known as Shazab in Saudi Arabia, is one of the famous culinary plants belonging to the Rutaceae family. It is commonly used in ethnomedicine in treating numerous diseases. This study was performed to characterize the essential oil isolated from Saudi species using a relatively new advanced headspace solid-phase microextraction technique. Following that, the antioxidant activity of the extracted oil was assessed using in vitro techniques such as the DPPH and nitric oxide scavenging tests, as well as the reducing power FRAP study and the molecular docking tool. The essential oil yield of the dried plant was 0.83% (v/w). Gas chromatography joined with a mass spectrometer was used to determine the chemical composition of the pale-yellow essential oil. Sixty-eight constituents were detected, representing 97.70% of the total oil content. The major constituents were aliphatic ketones dominated by 2-undecanone (37.30%) and 2-nonanone (20.00%), with minor constituents of mono and sesquiterpenoids chemical classes. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is one of the major causes of many contemporary diseases due to its ability to create a reactive oxygen species (ROS). Thus, molecular docking was used to confirm that some oil phytoconstituents have good docking scores compared to the standard antioxidant drug (Vitamin C), indicating great binding compatibility between the (NADPH) oxidase receptor site and the ligand. In conclusion, our findings suggest that the oil could be used safely and as a cost-effective remedy in treating various modern diseases caused by free radical formation.

QM/MM Modeling of the Flavin Functionalization in the RutA Monooxygenase.

Oxygenase activity of the flavin-dependent enzyme RutA is commonly associated with the formation of flavin-oxygen adducts in the enzyme active site. We report the results of quantum mechanics/molecular mechanics (QM/MM) modeling of possible reaction pathways initiated by various triplet state complexes of the molecular oxygen with the reduced flavin mononucleotide (FMN) formed in the protein cavities. According to the calculation results, these triplet-state flavin-oxygen complexes can be located at both re-side and si-side of the isoalloxazine ring of flavin. In both cases, the dioxygen moiety is activated by electron transfer from FMN, stimulating the attack of the arising reactive oxygen species at the C4a, N5, C6, and C8 positions in the isoalloxazine ring after the switch to the singlet state potential energy surface. The reaction pathways lead to the C(4a)-peroxide, N(5)-oxide, or C(6)-hydroperoxide covalent adducts or directly to the oxidized flavin, depending on the initial position of the oxygen molecule in the protein cavities.

The Influence of Exogenous Phenylalanine on the Accumulation of Secondary Metabolites in Agitated Shoot Cultures of Ruta graveolens L.

This study aimed to examine the influence of the addition of a precursor (phenylalanine) on the accumulation of secondary metabolites in agitated shoot cultures of Ruta graveolens. Cultures were grown on Linsmaier and Skoog (LS) medium, with plant growth regulators (0.1 mg/L α-naphthaleneacetic acid-NAA-and 0.1 mg/L 6-benzylaminopurine-BAP). Phenylalanine was added to the cultures at a concentration of 1.25 g/L after 4 and 5 weeks of growth cycles. Biomass was collected after 2, 4, and 7 days of precursor addition. Both control and experimental cultures had the same secondary metabolites accumulated. Using the HPLC method, linear furanocoumarins (bergapten, isoimperatorin, isopimpinellin, psoralen, and xanthotoxin), furoquinoline alkaloids (γ-fagarine, 7-isopentenyloxy-γ-fagarine, and skimmianine), and catechin were detected and quantified in the methanolic extracts. In turn, phenolic acids, such as gallic acid, protocatechuic acid, p-hydroxybenzoic acid, syringic acid, p-coumaric acid, and ferulic acid were detected in hydrolysates. The production of phenolic acids and catechin (1.5-fold) was significantly increased by the addition of precursor, while there was no significant effect on the production of coumarins and alkaloids. The highest total content of phenolic acids (109 mg/100 g DW) was obtained on the second day of phenylalanine addition (the fourth week of growth cycles). The dominant phenolic compounds were p-coumaric acid (maximum content 64.3 mg/100 g DW) and ferulic acid (maximum content 35.6 mg/100 g DW). In the case of catechins, the highest total content (66 mg/100 g DW) was obtained on the third day of precursor addition (the fourth week of growth cycles). This study is the first to document the effect of feeding the culture medium with phenylalanine on the accumulation of bioactive metabolites in in vitro cultures of R. graveolens.

Potential Protective Effects of Ruta Chalepensis L. Extracts Against Gentamicin-Induced Nephrotoxicity via Reduction on Apoptotic, DNA Damage and Oxidative Stress Markers in Mice.

In this study, the protective effects of Ruta chalepensis L. extracts on the extent of tissue damage in gentamicin-induced nephrotoxicity have been investigated. Ruta chalepensis L. extracts were prepared by subcritical water and ultrasound-assisted organic solvent extraction methods. Protective activity of Ruta chalepensis L. extracts on Gentamicin-induced nephrotoxicity is investigated by apoptotic, DNA damage, oxidative stress markers and evaluating histopathological in kidney tissue of mice. Gentamicin significantly increased Caspase-3 and -8 activities, NO levels, serum creatinine and BUN, while 8-OHdG and MDA levels were significantly decreased with Ruta chalepensis L. extract treatment. In addition, Ruta chalepensis L. extracts treatment significantly increased CAT and SOD activities. Histopathological alterations in Gentamicin group were significantly diminished by application of Ruta chalepensis L. extracts. These results suggest that treatment with Ruta chalepensis L. extracts may ameliorate renal dysfunction and structural damage through the reduction of oxidative stress and apoptosis in the kidney.

THE EFFECT OF RUTA GRAVEOLENS L. ETHANOLIC EXTRACTS ON SKIN ISOLATES OF STAPHYLOCOCCI AND PROPIONIBACTERIUM ACNES.

OBJECTIVE: The aim: To define antimicrobial properties of 50%, 70% and 90% ethanolic extracts of Ruta graveolens L. against macrolide resistant (MLS-resistant) skin isolates of staphylococci and Propionibacterium acnes, and to determine MIC and MBC of investigated extracts. PATIENTS AND METHODS: Materials and methods: Extracts were prepared by the method of maceration. Bacterial cultures were identified by biochemical microtests. Identification of MLS-resistance type was performed by using disc-diffusion method. The MIC and MBC were determined by serial two-fold dilution of ethanolic extracts of Ruta graveolens L. in MHB and HBB for staphylococci and P. acnes, respectively. Bacterial growth in each well was assayed by absorption at 495 nm, using a spectrophotometer SynergyTMHTX S1LFTA (BioTek Instruments, Inc., USA). RESULTS: Results: All ethanolic extracts of garden ruta were active against all staphylococci and P. acnes skin isolates and showed exclusively bactericidal activity (MBC/MIC ratios ranged from 1 to 2) against all investigated strains. 90% extract of Ruta graveolens L. showed better results than 50% and 70% extracts - average MIC and MBC concentations for P. acnes strains were 1.38«0.66 mg/mL and for staphylococcal strains average MIC was 2.1«1.16 mg/mL and MBC - 2.86«1.2 mg/mL. CONCLUSION: Conclusions: 50%, 70% and 90% ethanolic garden ruta extracts showed moderate antibacterial activity against main skin pathogens, responsible for acnes vulgaris development - S. epidermidis, S. aureus and P. acnes. No difference in susceptibility between resistance and sensitive strains of staphylococci and P. acnes indicate that acquired MLS-resistance of investigated skin isolates does not affect on the level of their sensitivity to ruta extracts.

The role of iron nanoparticles on morpho-physiological traits and genes expression (IRT1 and CAT) in rue (Ruta graveolens).

The iron nanoparticles with different physic-chemical properties induce inconsistent effects on various studied plant species. Thus, the effect of ferric oxide (Fe2O3) nanoparticles was compared with Fe2O3 microparticles and FeSO4complexes of EDTA for major physiological and gene expression in Rue (Ruta graveolens). Iron root content increased as Fe-MPs + EDTA ˂˂ Fe-NPs + EDTA˂ FeSO4 + EDTA. The shoot's iron remained unchanged or slightly increased under most of FeSO4 and Fe-MPs + EDTA treatments. Under Fe-NPs + EDTA treatment, 50 and 250 µM concentration decreased on shoot iron by 23.2% and 19.4% compared to control, respectively. But the shoot iron at 500 µM NPs was 28.2% higher than that of the control. A 46-58 fold lower Fe translocation was observed under Fe-NPs + EDTA than Fe-MPs + EDTA. The effect of Fe-NPs + EDTA was more significant on plant fresh and dry mass than the control. All treatments showed an increase in anthocyanin by 19-84% in leaves compared to the control. The Fe-NPs + EDTA and MPs + EDTA induced similar effects on enhanced growth parameters, total chlorophyll, catalase enzyme activity, gene, and reduced chlorophyll a/b and oxidants. Catalase enzyme activity in FeSO4 and MPs + EDTA was similar, and in Fe-NPs + EDTA treatments were influenced by coarse and fine regulation mechanisms, respectively. Iron MPs + EDTA had a more negative effect on IRT1 relative gene expression in roots as compared to other iron forms. The IRT1 relative gene expression in shoots was positively affected by 31-81% under all treatment types (except control and 250 µM Fe-NPs + EDTA, and 250 µM MPs + EDTA). These results could reveal the potential mechanism of plant response to nanoparticles.

Ruta graveolens, Peganum harmala, and Citrullus colocynthis methanolic extracts have in vitro protoscolocidal effects and act against bacteria isolated from echinococcal hydatid cyst fluid.

Echinococcosis is a common and endemic disease that affects both humans and animals. In this study, the in vitro activities of methanolic extracts of Ruta graveolens, Peganum harmala aerial parts, and Citrullus colocynthis seeds against protoscolosis and isolated bacterial strains from hydatid cysts were assessed using disc diffusion methods and Minimum Inhibitory Concentration (MIC). The chemical composition of three methanolic extracts was studied using LC-MS. After 3 h of exposure to 40 mg/mL R. graveolens extract, a tenfold protoscolocidal effect was seen when compared to the convintional medication (ABZ) for the same duration (P < 0.05). The bacteria listed below were isolated from hydatid cyst fluid collected from a variety of sick locations, including the lung and liver. Micrococcus spp., E. coli, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter amnigenus, Pseudomonas aeruginosa, Staphylococcus xylosus, and Achromobacter xylosoxidans are among the bacteria that have been identified. The most effective extract was R. graveolens, followed by P. harmala and C. colocynthis, according to the results of antibacterial activity using the disc diffusion method. R. graveolens extract had the lowest MIC values (less than 2 mg/mL) against all microorganisms tested. This shows that the R. graveolens extract has additional properties, such as the ability to be both scolocidal and bactericidal. Because these bacteria are among the most prevalent pathogenic bacteria that increase the risk of secondary infection during hydatid cysts, the results of inhibitory zones and MICs of the R. graveolens methanol extract are considered highly promising.

Antifungal, Repellency, and Insecticidal Activities of Cymbopogon distans and Ruta graveolens Essential Oils and Their Main Chemical Constituents.

Essential oils produced by Cymbopogon distans and Ruta graveolens with a similar Chinese name could be explained as book fragrance in the Chinese idiom 'shu xiang men di', namely, a wealthy intellectual family according to ancient Chinese. Therefore, volatile oils from these two plants and their main compounds were tested to explore their antifungal, repellent, and insecticide actions. In this study, the essential olis (EO) of C. distans exhibited significant antifungal activity against Rhizopus stolonifera (97 %), Mucor racemosus (97 %), and Trichoderma viride (84 %); its main compounds exhibit interesting activity, such as methyleugenol (87 %) and elemicine (85 %) against T. viride and butyl hydroxytoluene against M. racemosus (90 %) and R. stolonifera (95 %). The EO of R. graveolens and other major chemical constituents showed weak inhibitory effects against other fungi (Aspergillus flavus and Fusarium oxysporum). Then, EO (C. distans and R. graveolens) and its main compounds exhibited obvious repellent activity (more than 85 %) at a concentration of 16 nL/cm2 , which was consistent with the repellency of the positive control (DEET). In terms of insecticidal activity, the mortality of C. distans volatile oil against R. padi (56.4 %) was lower than that of R. graveolens volatile oil (92.4 %), and all compounds showed weak lethal effects. These results provide a natural substance for controlling fungi and insects when storing books and that can be used as a biological pesticide for industrial production. Through our study, the book fragrance in the Chinese idiom 'shu xiang men di' was speculated to be the EO odor of C. distans.

Phenolic Content and in Vitro Antioxidant, Anti-Inflammatory and antimicrobial Evaluation of Algerian Ruta graveolens L.

Plants constitute a valuable source of natural antioxidants such as polyphenols and are responsible for exhibiting many biologically significant functions. Ruta species including Ruta chalepensis L. and Ruta graveolens L. are widespread species in Algeria and are used as medicinal plants to treat various diseases; however, so far, most of the conducted studies focused on analyzing alkaloids and essential oils mostly on R. chalepensis. The aim of the present research is to investigate the phenolic profile of the aerial parts of Ruta graveolens L. from Algeria and assess its in vitro antioxidant, anti-inflammatory and antimicrobial properties. The total polyphenols and flavonoids were assessed using colorimetric methods, and the individual polyphenols were identified and quantified using HPLC-DAD-ESI-MS. The antioxidant activity was evaluated with DPPH and ß-carotene tests, and the anti-inflammatory activity with inhibition of bovine serum albumin denaturation and HRBC membrane stabilization methods. The results showed that Ruta graveolens extract is rich in phenolic compounds with a total phenol and flavonoid contents of 41.63±0.394 mg GAE/gE and 13.97±0.33 mg EQ/gE, respectively. Nine phenolic compounds were determined, including three phenolic acids and six flavonoids. Rutin was the major phenolic compound in Ruta graveolens (464.95 µg/g), followed by syringic acid (179.74 µg/g), and naringenin (109.78 µg/g). R. graveolens phenolic extract also showed good antioxidant activity with values of 0.77 mM TE/g DW and 0.37 mM ß-CE/g DW with DPPH and ß-carotene tests, respectively. For the anti-inflammatory activity, the highest tested concentration (200 µg/mL) gave 50.61 % of inhibition of the denaturation of albumin and 44.12 % of membrane stabilization. With regards to antimicrobial results, Staphylococcus aureus was the most sensitive bacteria with an inhibition zone of 14.37 mm and MIC value of 0.625 mg/mL, followed by Listeria monocytogenes (11.75 mm and MIC=1.25 mg/mL), and Escherichia coli (10.25 mm and MIC=1.25 mg/mL).

Biological toxicity of Ruta graveolens essential oil against three species of diptera Drosophila melanogaster, Culex pipiens and Culiseta longiareolata.

BACKGROUND & OBJECTIVES: Recently, the use of biodegradable and environment friendly plant-based bioinsecticides has received a great deal of attention from researchers to control insect disease vectors. The aim of this research is to determine the larvicidal efficacy of Ruta graveolens essential oil against third instar larvae of two species of mosquito (Culex pipiens and Culiseta longiareolata) and a biological model Drosophila melanogaster. METHODS: Culiseta longiareolata and Culex pipiens larvae were collected from untreated areas located in Tebessa and Drosophila melanogaster, the wild strain collected from rotten apples in the Tebessa region. Ruta graveolens essential oil has been tested at different concentrations between 2.5µ/mL and 140µL/mL against third instar larvae of the three species under standard laboratory conditions according to the recommendations from the Word Health Organization. The effects were examined on mortality, growth and the main components (proteins, carbohydrates, lipids). RESULTS: The essential oil showed larvicidal activity with LC50 and LC90 values (10.85µL/mL, 70.95µL/mL and 39.4µL/mL), (26µL/mL, 144.5µL/mL and 89.57µL/mL) against third instar larvae of Drosophila melanogaster, Culex pipiens and Culiseta longiareolata respectively. In addition, it disrupted the growth and several morphological malformations were observed. It also affected growth and the main components (proteins, carbohydrates, lipids). INTERPRETATION & CONCLUSION: The essential oil affected growth and energy reserves for all three species. The results indicated that the essential oil of Ruta graveolens has good potential as a source of natural larvicides.

Synergistic Effect of Biosynthesized Silver Nanoparticles and Natural Phenolic Compounds against Drug-Resistant Fish Pathogens and Their Cytotoxicity: An In Vitro Study.

Fish pathogens causing disease outbreaks represent a major threat to aquaculture industry and food security. The aim of the presented study is to develop safe and effective bioactive agents against two bacterial isolates: Aeromonas hydrophila and Pseudomonas fluorescens. We employed a broth microdilution method to investigate the antibacterial effect of biosynthesized silver nanoparticles (AgNPs); rutin, a natural flavonoid extracted from Ruta graveneoles; and heliomycin, a secondary metabolite produced by marine actinomycetes AB5, as monotherapeutic agents. Moreover, AgNPs in combination with rutin (AgNP + R) and heliomycin (AgNPs + H) were examined for their synergistic effect. The cytotoxic effect of individual bioactive compounds and in combination with AgNPs was investigated on epithelioma papulosum cyprini (EPC) fish cell lines. Individual treatment of AgNPs, rutin, and heliomycin exhibited a dose-dependent antimicrobial activity against A. hydrophila and P. fluorescens. Rutin minimum inhibitory concentration (MIC) showed the lowest cytotoxicity when tested on EPC cell lines, while heliomycin MIC was highly cytotoxic. Combined subtherapeutic doses of AgNPs + R and AgNPs + H displayed additive and synergistic effects against A. hydrophila and P. fluorescens, respectively, with improved results and relative safety profile. The study findings demonstrate that a combination of AgNPs and natural bioactive compounds may represent novel therapeutics fighting fish pathogens potentially affecting the fish farming industry.

Identification of Alkaloid Compounds Arborinine and Graveoline from Ruta angustifolia (L.) Pers for their Antifungal Potential against Isocitrate lyase (ICL1) gene of Candida albicans.

Candida albicans has been reported globally as the most widespread pathogenic species contributing candidiasis from superficial to systemic infections in immunocompromised individuals. Their metabolic adaptation depends on glyoxylate cycle to survive in nutrient-limited host. The long term usage of fungistatic drugs and the lack of cidal drugs frequently result in strains that could resist commonly used antifungals and display multidrug resistance (MDR). In search of potential therapeutic intervention and novel fungicidals, we have explored a plant alkaloids, namely arborinine and graveoline for its antifungal potential. Alkaloids belongs to Rutaceae family have been reported with numerous antimicrobial activities. In this study, we aimed to isolate and identify the antifungal active alkaloids of R. angustifolia and assess antifungal effect targeting C. albicans isocitrate lyase (ICL) gene which regulates isocitrate lyase, key enzyme in glyoxylate cycle contributing to the virulence potential of C. albicans. Alkaloids were extracted by bioassay guided isolation technique which further identified by TLC profile and compared with the standard through HPLC and NMR analysis. The antifungal activities of the extracted alkaloids were quantified by means of MIC (Minimum Inhibitory Concentration). The gene expression of the targeted gene upon treatment was analysed using RT-qPCR and western blot. Additionally, this study looked at the drug-likeness and potential toxicity effect of the active alkaloid compounds in silico analysis. Spectroscopic analysis showed that the isolated active alkaloids were characterized as acridone, furoquinoline, 4-quinolone known as arborinine and graveoline. Results showed that each compound significantly inhibited the growth of C. albicans at the dose of 250 to 500 µg/mL which confirm its antifungal activity. Each alkaloid was found to successfully downregulate the expression of both ICL1 gene CaIcl1 protein. Finally, ADMET analysis suggests a good prediction of chemical properties, namely absorption, distribution, metabolism, excretion and toxicity (ADMET) that will contribute in drug discovery and development later on.

Ruta Essential Oils: Composition and Bioactivities.

Ruta L. is a typical genus of the citrus family, Rutaceae Juss. and comprises ca. 40 different species, mainly distributed in the Mediterranean region. Ruta species have long been used in traditional medicines as an abortifacient and emmenagogue and for the treatment of lung diseases and microbial infections. The genus Ruta is rich in essential oils, which predominantly contain aliphatic ketones, e.g., 2-undecanone and 2-nonanone, but lack any significant amounts of terpenes. Three Ruta species, Ruta chalepensis L., Ruta graveolens L., and Ruta montana L., have been extensively studied for the composition of their essential oils and several bioactivities, revealing their potential medicinal and agrochemical applications. This review provides a systematic evaluation and critical appraisal of publications available in the literature on the composition and bioactivities of the essential oils obtained from Ruta species and includes a brief outlook of the potential applications of nanotechnology and chitosan-based products of Ruta essential oils.

Anti-MRSA Constituents from Ruta chalepensis (Rutaceae) Grown in Iraq, and In Silico Studies on Two of Most Active Compounds, Chalepensin and 6-Hydroxy-rutin 3',7-Dimethyl ether.

Ruta chalepensis L. (Rutaceae), a perennial herb with wild and cultivated habitats, is well known for its traditional uses as an anti-inflammatory, analgesic, antipyretic agent, and in the treatment of rheumatism, nerve diseases, neuralgia, dropsy, convulsions and mental disorders. The antimicrobial activities of the crude extracts from the fruits, leaves, stem and roots of R. chalepensis were initially evaluated against two Gram-positive and two Gram-negative bacterial strains and a strain of the fungus Candida albicans. Phytochemical investigation afforded 19 compounds, including alkaloids, coumarins, flavonoid glycosides, a cinnamic acid derivative and a long-chain alkane. These compounds were tested against a panel of methicillin-resistant Staphylococcus aureus (MRSA) strains, i.e., ATCC 25923, SA-1199B, XU212, MRSA-274819 and EMRSA-15. The MIC values of the active compounds, chalepin (9), chalepensin (10), rutamarin (11), rutin 3'-methyl ether (14), rutin 7,4'-dimethyl ether (15), 6-hydroxy-rutin 3',7-dimethyl ether (16) and arborinine (18) were in the range of 32-128 µg/mL against the tested MRSA strains. Compounds 10 and 16 were the most active compounds from R. chalepensis, and were active against four out of six tested MRSA strains, and in silico studies were performed on these compounds. The anti-MRSA activity of compound 16 was comparable to that of the positive control norfloxacin (MICs 32 vs 16 µg/mL, respectively) against the MRSA strain XU212, which is a Kuwaiti hospital isolate that possesses the TetK tetracycline efflux pump. This is the first report on the anti-MRSA property of compounds isolated from R. chalepensis and relevant in silico studies on the most active compounds.

Chalepin and Chalepensin: Occurrence, Biosynthesis and Therapeutic Potential.

Dihydrofuranocoumarin, chalepin (1) and furanocoumarin, chalepensin (2) are 3-prenylated bioactive coumarins, first isolated from the well-known medicinal plant Ruta chalepensis L. (Fam: Rutaceae) but also distributed in various species of the genera Boenminghausenia, Clausena and Ruta. The distribution of these compounds appears to be restricted to the plants of the family Rutaceae. To date, there have been a considerable number of bioactivity studies performed on coumarins 1 and 2, which include their anticancer, antidiabetic, antifertility, antimicrobial, antiplatelet aggregation, antiprotozoal, antiviral and calcium antagonistic properties. This review article presents a critical appraisal of publications on bioactivity of these 3-prenylated coumarins in the light of their feasibility as novel therapeutic agents and investigate their natural distribution in the plant kingdom, as well as a plausible biosynthetic route.