Novel Amides Derivative with Antimicrobial Activity of Piper betle var. nigra Leaves from Indonesia.

Piper betle var. nigra is a tropical plant closely related to the common piper. P. betle has also been dubbed a promising source of natural antioxidants in herbal health products, antibacterial, antifungal, antimalarial, cytotoxic activity against the cancer cell lines K562 and HL-60, and antileishmanial. The aim of this study to observation Antimicrobial activity and isolation of chemical compound. The antimicrobial activity of P. betle extract was performed by well diffusion method against two oral pathogenic bacteria (Streptococcus mutans and Streptococcus sanguinis) and opportunistic pathogenic yeast (Candida albicans). The inoculum (bacterial and yeast suspension) was prepared from a 24-h culture on NB for bacterial suspension and on TSB for yeast suspension. Extraction and isolation using various method of chromatography. Isolated compounds were characterized by spectroscopic means. Our study showed antimicrobial activity from crude ethanol extract of leaves P. betle L. var. nigra against two oral pathogenic bacteria and opportunistic pathogenic yeast with concentration 0.5% and 1%. The first report of two new amides derivatives, piperenamide A (1) and piperenamide B (2) in P. betle L. var. nigra.

Simultaneous Natural Deep Eutectic Solvent-Based Ultrasonic-Assisted Extraction of Bioactive Compounds of Cinnamon Bark and Sappan Wood as a Dipeptidyl Peptidase IV Inhibitor.

Cinnamon bark (Cinnamomum burmannii) and sappan wood (Caesalpinia sappan) have been reported to be beneficial for Type-2 Diabetes Mellitus (T2DM) and the combination is commonly used by Indonesian herbal industries. In the present study, the simultaneous extraction of bioactive compounds from both plants was conducted using natural deep eutectic solvent (NADES), their content analyzed using high-performance liquid chromatography (HPLC), and their dipeptidyl peptidase IV (DPP IV) inhibitory activity evaluated. An additional in silico molecular docking analysis was conducted to ensure their activity. The results showed that NADES (with a composition of choline chloride-glycerol) extraction from cinnamon and sappan wood had DPP IV inhibitory activity of 205.0 and 1254.0 µg/mL, respectively. Brazilin as a marker substance from sappan wood was responsible for the DPP IV inhibitory activity, while none of the marker substances chosen for cinnamon bark (trans-cinnamaldehyde, coumarin, and trans-cinnamic acid) were found to have significant DPP IV inhibitory activity. These results were confirmed by molecular docking conducted in brazilin, trans-cinnamaldehyde, coumarin, and trans-cinnamic acid.

Garcinia dulcis and Garcinia forbesii King fruit peel extract: Secondary metabolite composition, antioxidant, and elastase inhibitory activity evaluation.

Garcinia dulcis and Garcinia forbesii King are native plants from Indonesia and have tremendous potential as a source of raw medicines based on local wisdom. However, scientific data for strengthening pharmaceuticals are still limited. Therefore, it is necessary to conduct a study to strengthen and develop the potential of both plants using the approach of traditional medicine. This study aimed to explore the secondary metabolite composition and biological activity (antioxidant and antielastase) of both plants. Both samples were extracted using 70% ethanol and microwave-assisted extraction with a microwave power of 120 watts for 15 min. The extract obtained was then screened for phytochemicals using specific reagents. The total phenolic content (TPC) was determined using spectrophotometry with a 96-well microplate reader method. The total flavonoid content (TFC) was determined using the colorimetric method, whereas metabolite profiling analysis was conducted using the UPLC-QToF-MS/MS system. Meanwhile, biological activity was tested for antioxidant activity and antielastase as measured by a microplate reader 96-well spectrophotometry method at specific wavelengths. According to the results, G. dulcis and G. forbesii fruit peel extracts showed positive detection of particular secondary metabolites. TPC and TFC values were 13.98 ± 1.90 mg GAE/g and 10.33 ± 1.90 mg QE/g for G. dulcis and 11.98 ± 2.04 mgGAE/g and 1.96 ± 0.36 mgQE/g for G. forbesii. Metabolite profiling detected some compounds from G. dulcis, including ephedrannin B, hinokiflavone, mahuannin J, and candidate mass C9H12O8, and G. forbesii, including 5-Hydroxy-7,8,2'- trimethoxyflavone, lucialdehyde B, candidate mass C21H39NO4, candidate mass C14H10O6, and candidate mass C14H12O6. Meanwhile, the biological activities (antioxidant and antielastase) were 137.721 µg/mL and 108.893 µg/mL for G. dulcis and 481.948 µg/mL and 250.611 µg/mL for G. forbesii, respectively. Both plants showed different profiles of secondary metabolites and biological activities (antioxidant and antielastase) according to their respective characteristics.

NMR-based metabolic profiling and antioxidant activity of Eleutherine bulbosa bulb ethanol extract.

The metabolite profile of the ethanol extract from Eleutherine bulbosa was assessed using the 1D NMR approach amalgamated with 2D NMR. E. bulbosa, an indigenous plant found in Indonesia, possesses considerable promise as a raw material for medicinal products. We examined the antioxidant capacity of the E. bulbosa extract using DPPH and ABTS methods. The antioxidant evaluation demonstrated that the ethanol extract exhibits strong antioxidant properties. Furthermore, we have successfully identified thirteen metabolites in the ethanol extract without separation. These metabolites include eleutherol A, eleutherol B, eleutherol C, eleuthione B, eleuthione C, hongconin, karwinaphthol, isoeleuthoside C, quinic acid, chlorogenic acid, isoeleutherin, cyanidin-3-o-ß-glucopyranoside, and kadsuric acid. Metabolite Cyanidin-3-o-ß-glucopyranoside (4.89 mM ± 0.02) was the highest metabolite contained in this extract. This result showed that NMR methods could detect and measure metabolites in the E. bulbosa ethanol extract without requiring separation.

Natural Deep Eutectic Solvent based Ultrasound-assisted extraction: A green approach for extraction of sulfhydryl and mimosine from Leucaena leucocephala (Lam) de Wit seeds.

Leucaena leucocephala (Lam.) de Wit seeds, also known as river tamarind, contain sulfhydryl compounds that exhibit antioxidant effects. However, these seeds also possess a toxic effect from mimosine. In this study, the river tamarind seeds were extracted using a natural deep eutectic solvent (NADES) based UAE. Among six NADES compositions screened, choline chloride-glycerol (ChCl-Gly) and choline chloride-sucrose (ChCl-Suc) were selected to be further optimized using a Box-Behnken Design in the RSM. The optimization of total sulfhydryl content was performed in 17 runs using three variables, namely water content in NADES (39%, 41%, and 43%), extraction time (5, 10, and 15 min), and the liquid-solid ratio (3, 5, and 7 mL/g). The highest concentration of sulfhydryls was obtained from ChCl-Gly-UAE (0.89 mg/g sample) under the conditions of a water content in NADES of 41% (v/v) and a liquid-solid ratio of 3 mL/g for 15 min, followed by that of from ChCl-Suc-UAE extract under the conditions of water content in NADES of 43% (v/v) and the liquid-solid ratio of 3 mL/g for 10 min with total sulfhydryl level was 0.67 mg/g sample. The maceration method using 30% ethanol resulted in the lowest level of sulfhydryls with a value of 0.52 mg/g. The mimosine compounds obtained in the NADES-based UAE (ChCl-Suc and ChCl-Gly) extracts were 4.95 and 7.67 mg/g, respectively, while 12.56 mg/g in the 30% ethanol-maceration extract. The surface morphology of L. leucocephala seed before and after extraction was analyzed using scanning electron microscopy. Therefore, it can be concluded that the use of ChCl-Suc and ChCl-Gly in NADES-based UAE is more selective in attracting sulfhydryl compounds than that of 30% ethanol-maceration extraction.

Optimization of microwave-assisted extraction on polyphenol metabolite from Eleutherine bulbosa (Mill.) urb. bulbs using response surface methodology.

Eleutherine bulbosa bulbs, an endemic plant in Indonesia, have enormous potential as raw materials for pharmaceutical products. Therefore, it is necessary to strengthen and develop extraction methods that are easy, rapid, and efficient to enrich targeted secondary metabolites. This study aims to optimize the microwave-assisted extraction (MAE) method conditions for polyphenol metabolite from E. bulbosa bulbs. The MAE method (with different conditions) was applied to extract total polyphenol content (TPC) from E. bulbosa bulbs. TPC values were determined using a 96-well microplate reader spectrophotometry method and Folin-Ciocalteu reagent. The variables of MAE, as an experimental design-independent variable, were involved. The MAE method condition was optimized using response surface methodology (RSM) and Box-Behnken design based on the TPC value. The MAE condition was optimized with 60% ethanol, sample-solvent ratio of 1:10 g/mL, and 50% Watts of microwave power for 10 min. The quadratic regression analysis was achieved to predict the TPC value using the equation: TPC value = 28.63-5.545A +2.211B -0.741C +1.995D - 4.045AB +0.856AC -7.541BC +1.961CD -8.342A2-0.071B2 +1.840C2-1.535D2. For the scale-up confirmation test, a 50-g sample was used to prove the validity of the equation to predict the TPC value, yielding 35.33 ± 2.13 mg gallic acid equivalent/g samples. The optimum of the MAE condition recommended based on the results of RSM analysis can be applied directly to the enrichment of polyphenols metabolite constituent of E. bulbosa easily, cheaply, quickly, and efficiently.

Phytochemical, in vitro radical scavenging and in vivo oxidative stress analysis of peppermint (Mentha piperita L.) leaves extract.

This current work aims to determine phytochemicals, in vitro radical scavenging, and in vivo oxidative stress reduction activities of peppermint (Mentha piperita L.) ethanolic extract (PEE). The Clule method was used to determine the phytochemical content. An in vitro antioxidant with radical scavenging activity was measured using 2,2-diphenyl-1-picrylhydrazyl. An in vivo antioxidant with oxidative stress reduction was carried out for 10 days on 25 male Sprague-Dawley rats (divided into five groups). Every day, each group was given positive control, negative control, 5, 10, and 20 mg/200 gr of body weight (BW) of the extract. The blood plasma was taken for malondialdehyde analysis. A phytochemical identification of PEE revealed more compounds, such as flavonoids, alkaloids, steroids, essential oils, and tannin. PEE exhibits significant in vitro radical scavenging activity, with an IC50 value of 126.695 µg/mL. In the in vivo antioxidant with oxidative stress reduction experiments, 5 mg/200 gr BW was the most effective dose, as evidenced by a considerable drop in malondialdehyde level (0.312 nmol/mL) after and before treatment. In conclusion, PPE has the potential to be developed as a herbal antioxidant based on in vitro and in vivo test results.

New robustaflavone from Garcinia latissima Miq. leave and Its antibacterial activity.

Isolation and determination of antibacterial compounds from plants are essential to obtain a new antibacterial as a substitute for conventional resistant antibiotics. This study aims to isolate and identify a new robustaflavone as antibacterial activity from Garcinia latissima Miq. leave. In this study, the isolation process was carried out using column chromatography followed by preparative thin layer chromatography (TLC) based on the TLC profile. The fraction D was tested for anti-bacterial Bacillus subtilis using the TLC bioautography method. The isolates obtained were then identified using 1H-NMR, 13C-NMR, distortionless enhancement by polarization transfer, heteronuclear single quantum coherence, and heteronuclear multiple bond coherence. The Activity assay of the isolate was performed using the microdilution method. A pure compound obtained the result of the separation process with eluent n-hexane: Ethyl acetate (3:2) with Rf 0.6. This spot follows the spot in the contact bioautographic result of fraction D, the spot with Rf 0.6 gives an inhibition zone. After identifying and purifying the isolate were known as Robustaflavone, this compound has activity against B. subtilis with a (minimum inhibitory concentration) value of 2500 ppm. Robustaflavone successfully isolated and identified from G. latissima leave and its antibacterial activity.

In vitro studies on the cytotoxicity, elastase, and tyrosinase inhibitory activities of tomato (Solanum lycopersicum Mill.) extract.

Tomatoes (Solanum lycopersicum Mill.), a common vegetable in Indonesia, contain high levels of lycopene, which is good for the body. This research further investigates the activity of polar and nonpolar fractions of tomatoes as elastase and tyrosinase inhibitory, and cytotoxic agents. The extraction procedure used is maceration, fractionation through liquid-liquid fractionation, purification of phytochemical substances is achieved through the application of thin layer chromatography. Elastase and tyrosinase inhibitory activity was analyzed using spectrophotometry and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cytotoxic assay. The result showed that the extract yield was 0.004%. The percentage of polar fraction from the extract was 2.58%, while the nonpolar fraction was 0.69%. The elastase inhibitory activity of polar and nonpolar fractions of tomato extract is 87.21% ± 7.57% and 73.12% ± 7.44%, respectively, The elastase inhibitory activity of polar and nonpolar fractions of tomato extract is 87.21% ± 7.57% and 73.12% ± 7.44%, respectively. The fractions had higher the anti-elastase activity than the positive control quercetin (65.97% ± 3.00%). The tyrosinase inhibitory activity of polar and nonpolar fractions of tomato extract is 23.71% ± 7.91% and 41.16% ± 5.41% (kojic acid as standard is 65.07% ± 0.86%), respectively. The IC50 of the cytotoxic assay to NIH 3T3 mouse embryonic fibroblast cells of the polar and nonpolar fraction of tomato extract is 1820.90 µg/mL and 1643.86 µg/mL, respectively.

Dipeptidyl peptidase IV inhibition of phytocompounds from Artocarpus champeden (Lour.) Stokes: In silico molecular docking study and ADME-Tox prediction approach.

The present study examines the potential activity prediction based on free binding energy (ΔG) and interaction confirmation of phytocompounds from Artocarpus champeden (Lour.) Stokes with macromolecule protein receptor of dipeptidyl peptidase IV (DPP-IV) using in silico molecular docking studies and physicochemical and pharmacokinetic properties (ADME-Tox) prediction approaches. The active subsites of the DPP-IV receptor macromolecule protein Protein Data Bank (ID: 1 × 70) were docked using Autodock v4.2.6 (100 docking runs). A grid box of 52 × 28 × 26 Å points spaced by 0.37 Å was centered on the active site of x = 40.926 Å; y = 50.522 Å; z = 35.031 Å. For ADME-Tox prediction, Swiss ADME online-based application programs were used. The results show that 12 pythocompounds from A. champeden have the potential as DPP-IV inhibitors based on ΔG value and interaction conformation. There are five pythocompounds with lower ΔG values and inhibition constants than the native ligand and seven pythocompounds with ΔG values and inhibition constants close to the native ligand. The 12 compounds form an interaction conformation at the active subsites of the DPP-IV receptor. At the same time, the results of the ADME-Tox prediction analysis showed that the 12 compounds had different physicochemical and pharmacokinetic properties.

Diallyl Disulfide: A Bioactive Garlic Compound with Anticancer Potential.

Cancer is a life-threatening disease caused by the uncontrolled division of cells, which culminates in a solid mass of cells known as a tumor or liquid cancer. It is the leading cause of mortality worldwide, and the number of cancer patients has been increasing at an alarming rate, with an estimated 20 million cases expected by 2030. Thus, the use of complementary or alternative therapeutic techniques that can help prevent cancer has been the subject of increased attention. Garlic, the most widely used plant medicinal product, exhibits a wide spectrum of biological activities, including antibacterial, hypo-lipidemic, antithrombotic, and anticancer effects. Diallyl disulfide (DADS) is a major organosulfur compound contained within garlic. Recently, several experimental studies have demonstrated that DADS exhibits anti-tumor activity against many types of tumor cells, including gynecological cancers (cervical cancer, ovarian cancer), hematological cancers (leukemia, lymphoma), lung cancer, neural cancer, skin cancer, prostate cancer, gastrointestinal tract and associated cancers (esophageal cancer, gastric cancer, colorectal cancer), hepatocellular cancer cell line, etc. The mechanisms behind the anticancer action of DADS include epithelial-mesenchymal transition (EMT), invasion, and migration. This article aims to review the available information regarding the anti-cancer potential of DADS, as well as summarize its mechanisms of action, bioavailability, and pharmacokinetics from published clinical and toxicity studies.

Mitragyna Species as Pharmacological Agents: From Abuse to Promising Pharmaceutical Products.

Mitragyna is a genus belonging to the Rubiaceae family and is a plant endemic to Asia and Africa. Traditionally, the plants of this genus were used by local people to treat some diseases from generation to generation. Mitragyna speciosa (Korth.) Havil. is a controversial plant from this genus, known under the trading name "kratom", and contains more than 40 different types of alkaloids. Mitragynine and 7-hydroxymitragynine have agonist morphine-like effects on opioid receptors. Globally, Mitragyna plants have high economic value. However, regulations regarding the circulation and use of these commodities vary in several countries around the world. This review article aims to comprehensively examine Mitragyna plants (mainly M. speciosa) as potential pharmacological agents by looking at various aspects of the plants. A literature search was performed and information collected using electronic databases including Scopus, ScienceDirect, PubMed, directory open access journal (DOAJ), and Google Scholar in early 2020 to mid-2021. This narrative review highlights some aspects of this genus, including historical background and botanical origins, habitat, cultivation, its use in traditional medicine, phytochemistry, pharmacology and toxicity, abuse and addiction, legal issues, and the potential of Mitragyna species as pharmaceutical products.

In silico identification of natural products from Centella asiatica as severe acute respiratory syndromecoronavirus 2 main protease inhibitor.

Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) main protease (S-CoV-2 Mpro) is one of the main targets in designing antiviral against SARS-CoV-2. Centella asiatica contains several triterpenoids, polyacetylenes, and benzoic ester derivative with various biological activities including anti-inflammation and antiviral. Triterpenoids from C. asiatica could act as inhibitors of S-CoV-2 Mpro. The main objective of this study was to identify potential natural products from C. asiatica as S-CoV-2 Mpro inhibitor with better pharmacokinetic through in silico molecular docking method. : As much as 11 compounds from C. asiatica were docked with S-CoV-2 Mpro (PDB ID: 6LU7) using AutoDock v4.2.6. Pharmacokinetic parameters of these compounds were assessed using SwissADME (free access webserver). Molecular docking results of 11 natural products indicated that asiatate 6 and asiatate 10 have strong interaction with quite similar binding free energy compared to native ligand (‒9.00 and‒9.58 kcal/mol compared to ‒9.18 kcal/mol, respectively) with proper interaction to the catalytic dyad (His41 and Cys145). Pharmacokinetic analysis revealed that asiatate 4, asiatate 10, and asiatate 11 have poor pharmacokinetic properties. These results indicated that asiatate 6 could be recommended for further study as S-CoV-2 Mpro inhibitor.

Optimization of betaine-sorbitol natural deep eutectic solvent-based ultrasound-assisted extraction and pancreatic lipase inhibitory activity of chlorogenic acid and caffeine content from robusta green coffee beans.

Natural deep eutectic solvent (NADES) is an alternative approach in natural product extraction with various advantages, including low toxicity, biodegradable, and suitable phytochemical compounds in a wide range of polarity. Chlorogenic acid (CGA) and caffeine, a well-known compound in the coffee bean, have various potential health benefits. This study aims to optimize the betaine-sorbitol NADES-based ultrasound-assisted extraction (UAE) method of CGA and caffeine from Robusta green coffee beans and determine the inhibitory activity of robusta green coffee beans extract of the betaine-sorbitol NADES-UAE from the optimum condition on pancreatic lipase in vitro and in silico. The betaine-sorbitol NADES-UAE factors as experimental design variable parameters include betaine-sorbitol ratio (0.5:1.2, 1.25:1.2, and 2:1.2 mol), extraction time (10, 35, and 60 min), and solid-liquid ratio (1:10, 1:20, and 1:30 g/mL). Response surface methodology and Box-Behnken Design were used to optimize the extraction process. The response surface was calculated by using CGA and caffeine content as response values. CGA and caffeine content was determined by High-Performance Liquid Chromatography. Whereas in vitro lipase inhibitory activity assay examined by spectrophotometric measurement and in silico molecular docking analysis on PDB ID: 1LPB. According to the results, the optimum conditions of the betaine-sorbitol NADES-UAE have obtained the betaine-sorbitol ratio of 1.25: 1.2 mol, solid-liquid ratio of 1:30 mg/mL, and 60 min extraction time. Furthermore, obtained Robusta green coffee extract from the optimum condition of the betaine-sorbitol NADES-UAE showed high potential to inhibit lipase activity with IC50 of 18.02 µg/ml, comparable with IC50 of standard CGA (11.90 µg/ml) and caffeine (15.59 µg/ml), where potential interaction of both standards was confirmed using molecular docking analysis. Our finding demonstrated the optimum condition of the betaine-sorbitol NADES-UAE method for CGA and caffeine extraction and the potential pancreatic lipase inhibition activity from the Robusta green coffee bean.

Current State and Promising Opportunities on Pharmaceutical Approaches in the Treatment of Polymicrobial Diseases.

In recent years, the emergence of newly identified acute and chronic infectious disorders caused by diverse combinations of pathogens, termed polymicrobial diseases, has had catastrophic consequences for humans. Antimicrobial agents have been clinically proven to be effective in the pharmacological treatment of polymicrobial diseases. Unfortunately, an increasing trend in the emergence of multi-drug-resistant pathogens and limited options for delivery of antimicrobial drugs might seriously impact humans' efforts to combat polymicrobial diseases in the coming decades. New antimicrobial agents with novel mechanism(s) of action and new pharmaceutical formulations or delivery systems to target infected sites are urgently required. In this review, we discuss the prospective use of novel antimicrobial compounds isolated from natural products to treat polymicrobial infections, mainly via mechanisms related to inhibition of biofilm formation. Drug-delivery systems developed to deliver antimicrobial compounds to both intracellular and extracellular pathogens are discussed. We further discuss the effectiveness of several biofilm-targeted delivery strategies to eliminate polymicrobial biofilms. At the end, we review the applications and promising opportunities for various drug-delivery systems, when compared to conventional antimicrobial therapy, as a pharmacological means to treat polymicrobial diseases.

Review of Angiotensin-converting Enzyme Inhibitory Assay: Rapid Method in Drug Discovery of Herbal Plants.

The renin-angiotensin-aldosterone system is a signaling pathway which responsible in the blood pressure regulation. Angiotensin-converting enzyme (ACE) is one of the key elements responsible for the hypertensive mechanism. It converts angiotensin-I to angiotensin-II. The discovery history of the ACE inhibitory activity assay method has been through a long stage for decades and development continues until today. The ACE inhibitory activity has become an effective screening method in the search for new antihypertensive agents from herbal plants. Some of in vitro assay methods were used to examine the activity of ACE inhibitors based on the substrate usage, such as; Cushman and Cheung Method using a substrate hippuryl-histidyl-leucine (HHL), Holmquist method using a substrate furanacryloyl-tripeptide, Elbl and Wagner method using a substrate benzoil-[l-14C] glicyl-L-histidine-L-leucine, Carmel and Yaron method using a substrate o-aminobenzoylglycyl-p-nitrophenylalanilproline, and Lam method using 3-hydroxybutyrylglycyl-glycyl-glycine as substrate. Several different methods to measure the results of enzymatic reactions or separating substrate with products, including spectrophotometric, fluorometric, high-performance liquid chromatography, electrophoresis, and radiochemistry. Application of the test method for screening the ACE inhibitors activity and investigation of active compounds from natural products can be done easily with this method, it is very helpful in research because the results obtained are simple, accurate, and rapid.