In vitro Evaluation of Skin Related Enzyme Inhibitory Effect and Emulgel Formulation Development Studies of Onobrychis Argyrea subsp. Argyrea with Phytochemical Analysis.

Species of Onobrychis have been used to treat skin disorders such as wounds and cuts in folk medicine and Onobrychis argyrea subsp. argyrea (OA) commonly known as 'silvery sainfoin', is a member of this genus. In this study, it was aimed to investigate the skin-related biological activities and phytochemical characterization of OA. Moreover, an emulgel formulation was developed from the main methanolic extract of the plant (OAM). Initially, to identifiy of the active fractions, aerial parts of the plant material was extracted with methanol and fractionated by n-hexane, chloroform, ethyl acetate and n-butanol, respectively. Antioxidant activity was determined by CUPRAC, TOAC, FRAP and DPPH assays. Thereafter, the inhibition potential of OAM, novel formulation and all fractions was measured against elastase, collagenase, tyrosinase and hyaluronidase enzymes. OAM was analyzed and characterized by LC/MS-MS. The major bioactive flavonoids which are rutin and isoquercetin were measured and compared as qualitative and quantitative via high performance thin layer chromatography (HPTLC) analysis in OAM and fractions. The results showed that extracts of OA can be a potential cosmeceutical agent for skin related problems.

Antibacterial, antifungal and enzyme inhibitory effects of selected plants from Turkey.

In this study, antibacterial, antifungal, antihyaluronidase, anticollagenase and antielastase activity of Hypericum bithynicum, Malva neglecta, Morus alba, Rubus discolor, Sambucus ebulus and Smilax excelsa were investigated. Methanol extracts of M. neglecta and R. discolor and all extracts of H. bithynicum were more active against Staphylococcus epidermidis. Similarly, water extracts of M. alba and S. ebulus were more active against Streptococcus pneumonia. Additionally, S. ebulus and S. excelsa had prominent antifungal activity on Candida albicans. Besides, methanol extract of M. neglecta and n-hexane extract of H. bithynicum were determined to have significant antihyaluronidase activity. Only R. discolor showed significant antielastase effect.

Anti-Ageing Potential of S. euboea Heldr. Phenolics.

In recent years, the use of Sideritis species as bioactive agents is increasing exponentially. The present study aimed to investigate the chemical constituents, as well as the anti-ageing potential of the cultivated Sideritis euboea Heldr. The chemical fingerprinting of the ethyl acetate residue of this plant was studied using 1D and 2D-NMR spectra. Isomeric compounds belonging to acylated flavone derivatives and phenylethanoid glycosides were detected in the early stage of the experimental process through 2D-NMR techniques. Overall, thirty-three known compounds were isolated and identified. Some of them are reported for the first time not only in S. euboea, but also in genus Sideritis L. The anti-ageing effect of the ethyl acetate residue and the isolated specialized products was assessed as anti-hyaluronidase activity. In silico docking simulation revealed the interactions of the isolated compounds with hyaluronidase. Furthermore, the in vitro study on the inhibition of hyaluronidase unveiled the potent inhibitory properties of ethyl acetate residue and apigenin 7-O-ß-d-glucopyranoside. Though, the isomers of apigenin 7-O-p-coumaroyl-glucosides and also the 4'-methyl-hypolaetin 7-O-[6'''-O-acetyl-ß-d-allopyranosyl]-(1→2)-ß-d-glucopyranoside exerted moderate hyaluronidase inhibition. This research represents the first study to report on the anti-hyaluronidase activity of Sideritis species, confirming its anti-inflammatory, cytotoxic and anti-ageing effects and its importance as an agent for cosmetic formulations as also anticancer potential.

Antihyaluronidase and Alkaline Phosphatase (ALP) Activities of Medicinal Plants to Combat Echis carinatus Venom-Induced Toxicities.

Snakebite is one of the most neglected diseases of developing countries. Deaths due to snakebite envenoming are quite high in Pakistan, and many deaths are caused by Echis carinatus envenomation. Traditional use of medicinal plants against snakebites is a common practice in Pakistan due to countless benefits. The current study was performed with the objective to evaluate eighteen Pakistani medicinal plants inhibitory potential against hyaluronidase and alkaline phosphatase enzymes of Pakistani Echis carinatus venom. Hyaluronidase activity (0.2-1.6 mg/0.1 mL) and alkaline phosphatase activity (0.1-0.8 mg/0.1 mL) were measured in dose-dependent manner. Crude methanolic extracts of medicinal plants were used for in vitro investigation of their inhibitory activity against toxic enzymes. All active plants were fractioned using different solvents and were again analyzed for inhibitory activity of same enzymes. Results indicated all plants were able to neutralize hyaluronidase that Swertia chirayita (Roxb. ex Flem.) Karst., Terminalia arjuna Wight and Arn, Rubia cordifolia Thumb., and Matthiola incana (L.) R.Br. inhibited maximum hyaluronidase activity equivalent to standard reference (p > 0.5). Pakistani medicinal plants are dense with natural neutralizing metabolites and other active phytochemicals which could inhibit hyaluronidase activity of Pakistani Echis carinatus venom. Further advanced studies at molecular level could lead us to an alternative for envenoming of Pakistani Echis carinatus venom.

A Novel Apilic Antivenom to Treat Massive, Africanized Honeybee Attacks: A Preclinical Study from the Lethality to Some Biochemical and Pharmacological Activities Neutralization.

Massive, Africanized honeybee attacks have increased in Brazil over the years. Humans and animals present local and systemic effects after envenomation, and there is no specific treatment for this potentially lethal event. This study evaluated the ability of a new Apilic antivenom, which is composed of F(ab')2 fraction of specific immunoglobulins in heterologous and hyperimmune equine serum, to neutralize A. mellifera venom and melittin, in vitro and in vivo, in mice. Animal experiments were performed in according with local ethics committee license (UFRJ protocol no. DFBCICB072-04/16). Venom dose-dependent lethality was diminished with 0.25-0.5 µL of intravenous Apilic antivenom/µg honeybee venom. In vivo injection of 0.1-1 µg/g bee venom induced myotoxicity, hemoconcentration, paw edema, and increase of vascular permeability which were antagonized by Apilic antivenom. Cytotoxicity, assessed in renal LLC-PK1 cells and challenged with 10 µg/mL honeybee venom or melittin, was neutralized by preincubation with Apilic antivenom, as well the hemolytic activity. Apilic antivenom inhibited phospholipase and hyaluronidase enzymatic activities. In flow cytometry experiments, Apilic antivenom neutralized reduction of cell viability due to necrosis by honeybee venom or melittin. These results showed that this antivenom is effective inhibitor of honeybee venom actions. Thus, this next generation of Apilic antivenom emerges as a new promising immunobiological product for the treatment of massive, Africanized honeybee attacks.

Effect of phenolic compounds from the leaves of Psidium guajava on the activity of three metabolism-related enzymes.

Enzyme activity modulation by synthetic compounds provide strategies combining the inhibitory and therapeutic mode of action of the confirmed inhibitors. However, natural modulators could offer a valuable alternative for synthetic ones for the treatment of different chronic diseases (diabetes, hypertension, cancer); due to the numerous side effects of the latter. In vitro screening assays were conducted for Psidium guajava leaf methanolic extract against three metabolism-related enzymes; α-amylase, tyrosinase, and hyaluronidase. The obtained results showed that the examined extract retained weak and moderate multitarget inhibition against α-amylase, tyrosinase, and hyaluronidase, respectively; however, the leaf fractions exhibited stronger inhibitions for the three investigated enzymes. Fractionation of P. guajava leaf extract revealed that anthraquinones and ellagic acid are of the major active compounds with inhibitory activities for α-amylase, tyrosinase, and hyaluronidase. Kinetic studies showed that quinalizarin inhibition is competitive for both α-amylase and hyaluronidase, and ellagic acid inhibition for tyrosinase and hyaluronidase is competitive and un-competitive, respectively. The molecular docking studies of quinalizarin and ellagic acid with α-amylase, tyrosinase, and hyaluronidase showed high binding energies with different bonds stabilizing the ligand-protein complex. Compiling all obtained results led to conclude that both P. guajava leaf fractions, quinalizarin and ellagic acid, have multitarget activities with potential therapeutic applications in many metabolic disorders.

Hyaluronidase inhibitory saponins and a trypanocidal isoflavonoid from the aerial parts of Oxytropis lanata.

A chemical examination of an extract from the aerial part of Oxytropis lanata led to the isolation and identification of 36 compounds, including saponins, isoflavonoids, oxazoles, and glycosides. The three among them were previously unreported oleanane-type saponins. In trypanocidal screening, 5,7,4'-trihydroxyisoflavone showed inhibitory activity against Trypanosoma congolense (IC50 = 10.5 µM), the causative agent of African trypanosomosis in animals; this activity was similar to that of active compounds from the roots of this plant. O. lanata is known to be a traditional medicinal plant in Mongolia for the treatment of inflammatory diseases. The anti-hyaluronidase effect of saponins 3, 5, 8, and 9, (IC50 = 0.15-0.22 mM) was stronger than that of sodium cromoglicate, which was used as a reference drug (IC50 = 0.37 mM). The chemical structures of the new saponins were determined based on HRFABMS, 1H and 13C NMR, 1H-1H COSY, HMQC, HMBC, and ROESY spectroscopic data along with chemical procedures.

Enzyme inhibitory assessment of the isolated constituents from Plantago holosteum Scop.

Plants of the Plantago genus are widely used in Turkish folk medicine especially for the treatment of wound, abscess, and inflammation. The aqueous extract and five phenylethanoid glycosides acteoside (1), arenarioside (2), echinacoside (3), isoacteoside (4), and leucosceptoside A (5) isolated from the aerial parts and roots of Plantago holosteum Scop. (Plantaginaceae) were tested for their possible inhibitory activity against hyaluronidase, elastase, and collagenase, related to wound pathogenesis. Even though the aqueous extract prepared from the aerial parts (36.26%) and roots (47.01%) and the isolated compounds acteoside (29.13%), echinacoside (28.73%), and isoacteoside (31.69%) exerted a notable inhibition, arenarioside and leucosceptoside A were found inactive in the hyaluronidase enzyme inhibition test. Similar results were obtained from the collagenase enzyme inhibition test. The aqueous extract prepared from the aerial parts (31.09%) and roots (35.17%), echinacoside (25.13%), and isoacteoside (23.85%) exerted a notable inhibition in this test. However, none of the extracts and isolated compounds displayed elastase enzyme inhibitory activity. The experimental data demonstrated that P. holosteum displayed a remarkable enzyme inhibitory activity against hyaluronidase and collagenase. This paper is the first report regarding the in vitro enzyme inhibitory activity of P. holosteum.

Chemical Constituents, Antioxidant, Anti-MMPs, and Anti-Hyaluronidase Activities of Thunbergia laurifolia Lindl. Leaf Extracts for Skin Aging and Skin Damage Prevention.

This study aimed to investigate the potential usage of Thunbergia laurifolia Lindl. leaf extracts in the cosmetic industry. Matrix metalloproteinases (MMPs) and hyaluronidase inhibition of T. laurifolia leaf extracts, prepared using reflux extraction with deionized water (RE) and 80% v/v ethanol using Soxhlet's apparatus (SE), were determined. Rosmarinic acid, phenolics, and flavonoids contents were determined using high-performance liquid chromatography, Folin-Ciocalteu, and aluminum chloride colorimetric assays, respectively. Antioxidant activities were determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and linoleic acid-thiocyanate assays. MMP-1 inhibition was investigated using enzymatic and fluorescent reactions, whereas MMP-2, MMP-9, and hyaluronidase inhibition were investigated using gel electrophoresis. Cytotoxicity on human fibroblast cell line was also investigated. The results demonstrated that SE contained significantly higher content of rosmarinic acid (5.62% ± 0.01%) and flavonoids (417 ± 25 mg of quercetin/g of extract) but RE contained a significantly higher phenolics content (181 ± 1 mg of gallic acid/g of extract; p < 0.001). SE possessed higher lipid peroxidation inhibition but less DPPH• scavenging activity than RE. Both extracts possessed comparable hyaluronidase inhibition. SE was as potent an MMP-1 inhibitor as gallic acid (half maximal inhibitory concentration values were 12.0 ± 0.3 and 8.9 ± 0.4 mg/cm3, respectively). SE showed significantly higher MMP-2 and MMP-9 inhibition than RE (p < 0.05). Therefore, SE is a promising natural anti-ageing ingredient rich in rosmarinic acid and flavonoids with antioxidant, anti-hyaluronidase, and potent MMPs inhibitory effects that could be applied in the cosmetic industry.

Anti-inflammatory and anti-nociceptive property of Capparis tomentosa Lam. root extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Capparis tomentosa Lam. root is one of the commonly used traditional medicines for the treatment of pain and inflammatory conditions by the Kunama ethnic group of Ethiopia. Yet, its ethnomedicinal claim has not been scientifically investigated. AIM OF THE STUDY: This study aims to evaluate the anti-inflammatory and anti-nociception activities of root extracts of Capparis tomentosa Lam. MATERIALS AND METHODS: The anti-inflammatory activity of the 70% ethanolic extracts and solvent fractions was determined using invitro hyaluronidase inhibition assay and in vivo carrageenan-induced paw edema in Swiss albino mice model. The anti-nociception effect was determined using the acetic acid-induced writhing model and tail immersion model in Swiss albino mice. RESULTS: The root extract of Capparis tomentosa has exhibited significant hyaluronidase inhibition activity which was comparable with the reference drug, indomethacin. Significant inhibition of carrageenan-induced paw edema was observed in all the test sample treated mice compared to the negative control with the highest activity observed from butanol fraction which was equivalent to the effect of acetylsalicylic acid. All the root extract and solvent fractions have possessed a significant and dose-dependent abdominal writhing inhibition with the highest effect observed in the aqueous fraction. Additionally, the crude extract was found better in increasing the reaction latency of the mice to the hot water stimulus than the solvent fractions which were significant after 60-120 min compared to the distilled water treated group. The root extract and solvent fractions were also found safe at a single oral dose of 2 mg/g in Swiss albino mice. CONCLUSION: The findings of the present study may partially support the folkloric use of Capparis tomentosa root for the treatment of pain and inflammation.

Acylated iridoid glycosides with hyaluronidase inhibitory activity from the rhizomes of Picrorhiza kurroa Royle ex Benth.

Seven new acylated iridoid glycosides, picrorhizaosides A-G (1-7), were isolated from the methanol extract of the rhizomes of Picrorhiza kurroa Royle ex Benth. (Plantaginaceae), in addition to six known iridoid glycosides (8-13). The structures of these new iridoids, including their stereochemistry, were determined based on chemical and physicochemical evidence derived from NMR and MS analysis. Of the isolates, picrorhizaosides D (4, IC50 = 43.4 µM) and E (5, 35.8 µM); picrosides I (8, 60.7 µM), II (9, 22.3 µM), and IV (11, 59.2 µM); and minecoside (13, 57.2 µM), exhibited a similar or stronger hyaluronidase inhibitory activity than those of the antiallergic medicines disodium cromoglycate (64.8 µM), ketotifen fumarate (76.5 µM), and tranilast (227 µM).

Effect of phenolic compounds from the rind of Punica granatum on the activity of three metabolism-related enzymes.

Enzyme activity modulation by synthetic compounds provide strategies combining the inhibitory and therapeutic mode of action of the confirmed inhibitors. However, natural modulators could offer a valuable alternative for synthetic ones for the treatment of different chronic diseases (diabetes, hypertension, cancer) due to the numerous side effects of the latter. In vitro screening assays were conducted for Punica granatum rind methanolic extract against three metabolism-related enzymes: α-amylase, tyrosinase, and hyaluronidase. The obtained results showed that the examined extract retained high multitarget inhibition with inhibition percentages 31.5 ± 1.3%, 75.9 ± 4.7%, and 68.5 ± 5.3% against α-amylase, tyrosinase, and hyaluronidase, respectively. Bioguided fractionation of P. granatum rind extract revealed that quercetin is the major active compound with inhibitory activities: 54.3 ± 2.7%, 94.2 ± 3.5%, and 90.9 ± 2.7% against α-amylase, tyrosinase, and hyaluronidase, respectively. Kinetic studies of enzymes showed that quercetin inhibition was noncompetitive, uncompetitive, and competitive for α-amylase, tyrosinase, and hyaluronidase, respectively. The molecular docking of quercetin with α-amylase and hyaluronidase showed high binding energy with different bonds stabilizing the ligand-protein complex. Compiling all obtained results led to conclude that both P. granatum rind extract and quercetin have multitarget activities with potential therapeutic applications in many metabolic disorders.

Combining in silico and in vitro approaches to identification of potent inhibitor against phospholipase A2 (PLA2).

Phospholipase A2 (PLA2) is the main constituent of snake venom. PLA2 enzymes catalyze the Ca2+ dependent hydrolysis of 2-acyl ester bonds of 3-sn-phospholipids, releasing fatty acids and lysophospholipids. Inside the body of the victim, PLA2 from snake venom induces either direct or indirect pathophysiological effects, including anticoagulant, inflammatory, neurotoxic, cardiotoxic, edematogenic, and myotoxic activities. Therefore, there is a need to find the potential inhibitors against PLA2 responsible for snakebite. In this study, we employed in silico and in vitro methods to identify the potential inhibitor against PLA2. Virtual screening and molecular docking studies were performed to find potent inhibitor against PLA2 using Traditional Chinese Medicine Database (TCM). Based on these studies, Scutellarin (TCM3290) was selected and calculated by density functional theory calculation at B3LYP/6-31G**++ level to explore the stereo-electronic features of the molecule. Further, molecular docking and DFT of Minocycline was carried out. Quantum polarized ligand docking was performed to optimize the geometry of the protein-ligand complexes. The protein-ligand complexes were subjected to molecular dynamics simulation and binding free energy calculations. The residence time of a protein-ligand complex is a critical parameter affecting natural influences in vitro. It is nonetheless a challenging errand to expect, regardless of the accessibility of incredible PC assets and a large variety of computing procedures. In this metadynamics situation, we used the conformational flooding technique to deal with rank inhibitors constructions. The systematic free energy perturbation (FEP) protocol and calculate the energy of both complexes. Finally, the selected compound of TCM3290 was studied in vitro analysis such as inhibition of PLA2 activity, hyaluronidase activity and fibrinogenolytic activity. The TCM3290 had a more binding affinity compare to Minocycline, and interacted with the key residues of TYR63 and GLY31. DFT represented the highest HOMO and LUMO energy of 0.15146 eV. MD simulation with 100 ns proved that an inhibitor binding mode is more stable inside the binding site of PLA2. In vitro analysis shows that TCM3290 significantly neutralized by PLA2. The above observations confirmed that Scutellarin (TCM3290) had a potent snake venom neutralizing capacity and could hypothetically be used for therapeutic drives of snakebite envenomation.

Efficient identification of Acetylcholinesterase and Hyaluronidase inhibitors from Paeonia parnassica extracts through a HeteroCovariance Approach.

ETHNOPHARMACOLOGICAL RELEVANCE: On the basis of the relevant reference in the poem Theriaca of the ancient Greek physician Nicander and its traditional use, Paeonia parnassica was selected for the evaluation of two extracts obtained from the roots and aerial parts to inhibit hydrolytic enzymes involved in snake envenomation. The secondary metabolites which contribute to these activities were detected through a novel HeteroCovariance NMR based approach. Afterwards these ingredients were isolated, identified and evaluated for their inhibitory potency. AIM OF THE STUDY: The identification of acetylcholinesterase and hyaluronidase inhibitors from Paeonia parnassica extracts was used as a case study for the introduction of a recently developed methodology to evaluate ethnopharmacological data and exploit them for the discovery of bioactive natural compounds. This process is based on the fractionation of the selected extracts and the simultaneous phytochemical analysis and biological assessment of the resulting fractions, which permits the rapid detection of the specified secondary metabolites prior to any laborious and time-consuming purification. MATERIALS AND METHODS: The roots and aerial parts of P. parnassica were extracted using methanol: water 50:50 and the two resulted extracts were fractionated by Centrifugal Partition Chromatography. The obtained fractions were evaluated in-vitro for their ability to inhibit acetylcholinesterase and hyaluronidase enzymes and their 1H NMR spectra were recorded. The biological activity was statistically correlated with the spectral data through the HeteroCovariance Approach (HetCA). Finally the purification, identification and biological evaluation of targeted secondary metabolites were carried out. RESULTS: The general chemical structures and some explicit secondary metabolites which contribute (e.g. gallotannins, gallic acid derivatives) or not (characteristic "cage-like" monoterpenes of the genus, glycosylated flavonoids) to the anti-acetylcholinesterase and anti-hyaluronidase activities were detected through HetCA. The consequent isolation and biological evaluation of targeted compounds were performed in order to validate the effectiveness and precision of the methodology. This procedure revealed the most active ingredients of both extracts obtained from roots and aerial parts against the above mentioned biological targets, as well as other compounds possessing moderate activity. CONCLUSIONS: The results of this study contributed to the verification of the ancient text Theriaca regarding the use of Paeonia parnassica to treat the snake bite symptoms. Furthermore, the ingredients of the Paeonia parnassica extracts, which were responsible for their anti-cholinesterase and anti-hyaluronidase activities, were determined applying a HetCA methodology before their isolation. Therefore, the current work provides clear evidence that HetCA could consist an efficient tool for the exploitation of traditional medicine information in order to discover bioactive natural compounds and develop new pharmacotherapies which serve the needs of contemporary medicine.

Anti-inflammatory activity of saponins from roots of Impatiens parviflora DC.

Two triterpene saponins (IPS-1, IPS-2) for the first time were isolated from the roots of Impatiens parviflora DC. (Balsaminaceae). Their anti-inflammatory activity was evaluated by means of two in vitro models: anti-hyaluronidase and anti-denaturation assays. Both saponins were shown to be potent hyaluronidase inhibitors that affect the enzyme in a dose-dependent manner. The anti-hyaluronidase effect of IPS-2 (IC50 = 286.7 µg/mL) was higher than that of the reference drug: escin (IC50 = 303.93 µg/mL). Both saponins protected bovine serum albumin from heat-induced denaturation in a dose-dependent manner. IPS-1 demonstrated higher anti-denaturation effect (IC50 = 86.7 µg/ml) than IPS-2 (IC50 = 109.76 µg/mL) or the standard drug: acetylsalicylic acid (IC50 = 262.22 µg/mL). In conclusion, potent activity of IPS-1, IPS-2 in both in vitro assays shows that saponins from I. parviflora have anti-inflammatory activity. The obtained results allow to suggest that such compounds may be beneficial in inflammatory conditions, especially associated with excessive degradation of hyaluronic acid.

Inhibitory activity of Podospermum canum and its active components on collagenase, elastase and hyaluronidase enzymes.

Present study is aimed to investigate in vitro inhibitory effects of the extract prepared from the aerial parts of Podospermum canum (syn: Scorzonera cana var. jacquiniana) (Asteraceae) on hyaluronidase, collagenase, and elastase enzymes using a bioassay-guided fractionation. Inhibitory effects of the extract, sub-extracts, fractions obtained by column chromatography, and isolated compounds on collagenase, elastase, and hyaluronidase were performed by using in vitro enzyme inhibitory assays based on spectrophotometric evaluation. The methanolic extract obtained from P. canum exhibited strong inhibitory activities on elastase and collagenase while the insignificant activity was observed on hyaluronidase. Through bioactivity-guided fractionation, the ethyl acetate and remaining water sub-extracts obtained from the methanolic extract displayed significant inhibitory activities on collagenase and elastase, while petroleum ether and chloroform extracts did not show any inhibitory activity. Eleven known compounds: arbutin, 6́-O-caffeoylarbutin, cichoriin, 3,5-dicaffeoylquinic acid methyl ester, apigenin 7-O-ß-glucoside, luteolin 7-O-ß-glucoside, apigenin 7-O-ß-rutinoside, isoorientin, orientin, vitexin, procatechuic acid, and new compound 4-hydroxy-benzoic acid 4-(6-O-α-rhamnopyranosyl-ß-glucopyranosyl) benzyl ester have been obtained from ethyl acetate sub-extract. Results of the present study have revealed that apigenin 7-O-ß-glucoside, luteolin 7-O-ß-glucoside, apigenin 7-O-ß-rutinoside, and isoorientin showed potent enzyme inhibitory activities. However, methanolic extract of P. canum displayed a greater inhibitory activity than fractions and isolated compounds both on collagenase and elastase.

Investigation of anti-inflammatory and anti-cancer activity of Justicia adathoda metabolites.

To analyse metabolic compounds of Justicia adathoda to evaluate against pathogens, inflammation and cervical cancer. The investigation exposed that the extracts of Justicia adathoda have potent metabolic to eradicate the human diseases. The antibacterial, tumorolytic and anti-inflammatory activity of ethyl acetate and aqueous extracts of Justicia adathoda (leaves) were assessed. In vitro anti-inflammatory activity was assessed by standard procedures. Justicia adathoda metabolic exhibit anticancer activity in human cervical cancer cell line (HeLa) (in vitro) analysis. Flavonoids, saponins, alkaloids, amino acids, tannins and terpenoids were present in both the extracts. The active components present in the extracts were found to be amino acids, alkaloids, lipids and triterpenoids which have antibacterial activity shows inhibition against Salmonella and Escherichia coli. Justicia adathoda possesses significant anti-inflammatory activity and it was confirmed by in-vitro analysis. The anticancer activity was found effective in human cervical cancer cell line (HeLa) (in-vitro) analysis. From the investigation could conclude that the metabolic compounds Justicia adathoda is effective against Anti-inflammation and ethyl acetate extract of Justicia adathoda are effective for Cancer.

Inhibitory activity of Scorzonera latifolia and its components on enzymes connected with healing process.

ETHNOPHARMACOLOGICAL RELEVANCE: Scorzonera latifolia (Fisch. & Mey.) DC. (Asteraceae) grows naturally in Eastern Anatolia, northeastern Iran, and Caucasus. Latex of S. latifolia roots is used in Turkish folk medicine for its analgesic effects, externally to cure infertility in women, and internally as an antihelmintic. The milk obtained from the stem of the Scorzonera species is used for wound healing activity. Antinociceptive, anti-inflammatory, wound-healing, antioxidant, and antimicrobial activities have previously been reported for S. latifolia. AIM OF THE STUDY: A methanol extract of the aerial parts of Scorzonera latifolia that had been shown to possess wound-healing activity, was used to elucidate the possible mechanism of the wound-healing activity and to identify the compound(s) responsible for the effect by means of bioassay-guided fractionation. MATERIALS AND METHODS: The wound-healing activity potential of methanol extract of S. latifolia was detected by evaluating the inhibitory activity on the collagenase, hyaluronidase and elastase, which play important roles in the wound-healing process. Succesive fractionation of the methanol extract using petroleum ether, chloroform, ethyl acetate, respectively, and the residual wateryielded four respective fractions. The ethyl acetate part, which was determined as the most active fraction, was selected for further separation using chromatographic techniques. RESULTS: Ethylacetate fraction exhibited significant inhibitory activities on collagenase and elastase. Chromatographic separation of the ethylacetate extract yielded an active subfraction, from which was used to isolate quercetin-3-O-ß-apiofuranosyl-(1'''→2'')-ß-D-glucopyranoside (1), quercetin-3-O-α-rhamnopyranosyl-(1→6)-ß-D-galactopyranoside (2), isoorientin (3), and 7-methylisoorientin (4). Of the compounds tested, 7-methylisoorientin (4) exerted inhibitory activity on collagenase and elastase, while quercetin-3-O-ß-apiofuranosyl-(1'''→2'')-ß-glucopyranoside (1) inhibited collagenase only. None of the fractions, or isolated compounds showed any inhibitory effect on hyaluronidase. It must be mentioned, that in vitro tests showed that compounds 1-4 inhibit the collagenase and elastase and could help wound-healing process. However, the inhibititory effect of the methanol extract appears to be greater than that of both of the ethylacetate fraction, subfraction G and the isolated compounds, which suggest that a synergistic interaction of several compounds could be responsible for the wound-healing activity of the aerial parts of S. latifolia.

Effect of diclofenac etalhyaluronate (SI-613) on the production of high molecular weight sodium hyaluronate in human synoviocytes.

BACKGROUND: We have reported that a single intra-articular injection of diclofenac etalhyaluronate (SI-613) exerted a potent and long-lasting analgesic effect in experimental arthritis models. In the present study, we investigated the effect of SI-613 on the production of high molecular weight hyaluronic acid (HMW-HA) in synoviocytes from osteoarthritis (OA) patients and compared its efficacy with that of hyaluronic acid (HA). METHODS: We compared the effect of SI-613, HA, and diclofenac sodium (DF-Na) on high molecular weight HA production by human synoviocytes. RESULTS: SI-613 and exogenous HA induced the production of high molecular weight HA in synoviocytes from OA patients, whereas DF-Na had no effect. The molecular weight of newly produced HA was about 1000 kDa in the HA-treated synoviocytes and much higher than 2400 kDa in the SI-613-treated cells. The effect of the mixture of HA and DF-Na was similar to that of HA alone in that the molecular weight of newly produced HA was around 1000 kDa. SI-613 significantly suppressed hyaluronidase 2 (HYAL2) mRNA expression and significantly enhanced hyaluronan synthase 2 (HAS2) mRNA expression. HA had no effect on the expression levels of HYAL and HAS. CONCLUSION: The present results clearly demonstrate that SI-613 induces the production of high molecular weight HA in synoviocytes from OA patients, suggesting the long-lasting analgesic and disease modifying effect of SI-613 for OA. Taken together with the anti-inflammatory and analgesic effects we recently reported for the intra-articular administration of SI-613 to experimental animal models, SI-613 holds great promise for the treatment of knee osteoarthritis.

Hyal-1 inhibitors from the leaves of Phyllanthus muellerianus (Kuntze) Excell.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves and twigs from Phyllanthus muellerianus Kuntze Excell are known to exert anti-inflammatory and antipyretic properties as well as wound healing properties. During a wide screening for human hyaluronidase-1 inhibitors from natural sources leaf extracts from P. muellerianus turned out to show basic anti-hyaluronidase activity. A detailed investigation of this effect should rationalize the potential anti-inflammatory activity of the extract for improved wound healing. AIM OF THE STUDY: The following study aimed to characterize the anti-Hyal-1 activity of the extract from P. muellerianus and to pinpoint the responsible natural products responsible for this bioactivity. MATERIALS AND METHODS: Using cell surface displayed human Hyal-1 on Escherichia coli, the activity of inhibitors was determined by the stains-all assay method. A hydroalcoholic extract PWE from P. muellerianus was subjected to bioactivity-guided fractionation. Active compounds were characterized by means of mass spectrometry and NMR. RESULTS: PWE exerts a concentration dependent inhibition of Hyal-1 with an IC50 of 80 µg/mL. Bioassay-guided fractionation revealed 13 compounds from the two most active fractions, mainly ellagitannins and flavonoid glycosides. The most activeHyal-1 inhibitor was found to be the ellagitannin chebulanin 10 (IC50 132 µM). This represents the first description of chebulanin in P. muellerianus. CONCLUSIONS: The hydroalcoholic extract of P. muellerianus, as well as several subfractions obtained during bioassay-guided fractionation showed strong activity against Hyal-1. The main activity can be correlated to the ellagitanin chebulanin. Additionally, also synergistic effects are observed, indicating that the traditional use of aqueous extracts of P. muellerianus is justified, rather than the use of the isolated tannins. The traditional use of the plant as an anti-inflammatory agent for improved wound-healing can be rationalized by the anti-Hyal-1 activities of its constituents.