Elastase Inhibitory Activity Research Articles

화장품 소재로서의 꽃 10 종 에탄올추출물 생리활성 특성연구

Background: We investigated the antioxidant, anti-wrinkles, whitening, and moisturizing properties and amounts of phenolic compounds of ethanol extracts from flowers of 10 resource plants from Namwon and Mt. Jiri., Korea.<BR>Methods and Results: We measured antioxidant efficacy based on the total polyphenol, and total flavonoid content, and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. We evaluated the inhibitory effect on melanin synthesis and tyrosinase activity for the whitening effect. Furthermore, we analyzed the elastase and matrix metalloproteinase-1 (MMP-1) inhibition activity for anti-wrinkle capacity. To evaluate the moisturizing effect, we examined hyaluronan synthase (HAS) mRNA expression. In addition, the 19 phenolic compounds were detected using high performace liquid chromatography (HPLC). Among the 10 flowers, the antioxidant effect was high in the order of Rosa multiflora, Nelumbo nucifera, and Elsholtzia splendens. Whitening effect was high in the order of N. nucifera, R. multiflora, and Dendranthema zawadskii. As for the anti-wrinkle property, N. nucifera was the most effective followed by R. multiflora. Taraxacum coreanum was the best for moisturizing effect, followed by D. zawadskii, and E. splendens. Seven phenolic compounds were detected in the extracts of the 10 flowers.<BR>Conclusions: Overall, the extracts of five flowers extracts showed strong potential as antioxidant, whitening, anti-wrinkle, and moisturizing functional cosmetic agents.

Physicochemical Properties of Mucin Extracted from &lt;i&gt;Misgurnus mizolepis&lt;/i&gt; Epidermal Mucus

The purpose of this study was to investigate the extraction yield and physicochemical characteristics of mucin from the epidermal mucus of Misgurnus mizolepis. Ninety percent ethanol produced the highest yield of mucin. At higher concentrations, mucin had more elastic properties, as revealed by dynamic viscoelasticity measurements. Differential scanning calorimetry (DSC) demonstrated that mucin is a thermostable glycoprotein. Denaturation enthalpy increased as mucin concentration increased. Mucin inhibited the growth of both Escherichia coli and Bacillus cereus, but had better antimicrobial activity in the former. Mucin also exhibited antioxidant activity, equivalent to 66.64% of that of L-cysteine. Mucin inhibited elastase activity and MMP-1 & MMP-2 expression activity, and induced hyaluronic acid expression. It is indicating that mucin is effective in moisturizing and anti-aging the skin. Thus, mucin from the epidermal mucus of Misgurnus mizolepis could have great potential as a non-food material in various industries.

Anti-inflammatory effects of troxerutin are mediated through elastase inhibition

Context Obesity is a chronic low-grade inflammatory state associated with immune cell infiltration into the adipose tissue (AT). We hypothesize that the anti-obesity and anti-inflammatory effects of troxerutin (TX) are mediated through inhibition of elastase. Objective To determine the inhibitory effect of TX on elastase in vitro and in tumor necrosis factor alpha (TNFα) induced 3T3-L1 adipocytes and the molecular interaction of TX with human neutrophil elastase (HNE). Materials and Methods Differentiated 3T3-L1 adipocytes were pretreated with TX, elastatinal (ELAS) or sodium salicylate (SAL) before exposure to TNFα. Lipid accumulation, reactive oxygen species (ROS) generation and oxidant-antioxidant balance were examined. The mRNA and protein expression of TNFα, interleukin-6, monocyte chemoattractant protein-1, adiponectin, leptin, resistin, chemerin, and elastase were analyzed. Elastase inhibition by TX and ELAS in a cell free system and docking studies for HNE with TX and ELAS were performed. Results TX, ELAS or SAL pretreatment had lowered lipid droplets formation and TG content. TX suppressed ROS generation, oxidative stress and improved antioxidant status. The expression of inflammatory cytokines and elastase was downregulated while that of adiponectin was upregulated by TX. The concentration required to produce 50% inhibition in vitro (IC50) was 11.5 μM for TX and 16.9 μM for ELAS. TX showed hydrogen bonding and hydrophobic interactions with elastase. Discussion TNFα induces inflammation of 3T3-L1 cells through elastase activation. TX inhibits elastase activity, downregulates expression and binds with elastase. Conclusion The antioxidant and anti-inflammatory activities of TX in AT could be of relevance in the management of obesity.

Protection against UVB-Induced Photoaging by Nypa fruticans via Inhibition of MAPK/AP-1/MMP-1 Signaling.

Ultraviolet B (UVB) irradiation is major causative factor in skin aging. The aim of the present study was to investigate the protective effect of a 50% ethanol extract from Nypa fruticans (NF50E) against UVB-induced skin aging. The results indicated that NF50E exerted potent antioxidant activity (IC50 = 17.55 ± 1.63 and 10.78 ± 0.63 μg/mL for DPPH and ABTS-radical scavenging activity, respectively) in a dose-dependent manner. High-performance liquid chromatography revealed that pengxianencin A, protocatechuic acid, catechin, chlorogenic acid, epicatechin, and kaempferol were components of the extract. In addition, the extract exhibited elastase inhibitory activity (IC50 = 17.96 ± 0.39 μg/mL). NF50E protected against UVB-induced HaCaT cell death and strongly suppressed UVB-stimulated cellular reactive oxygen species generation without cellular toxicity. Moreover, topical application of NF50E mitigated UVB-induced photoaging lesions including skin erythema and skin thickness in BALB/C mice. NF50E treatment inhibited UVB-induced collagen degradation as well as MMP-1 and IL-1β expressions and significantly stimulated SIRT1 expression. Furthermore, the extract treatment markedly suppressed the activation of NF-κB and AP-1 (p-c-Jun) by deactivating the p38 and JNK proteins. Taken together, current data suggest that NF50E exhibits potent antioxidant potential and protection against photoaging by attenuating MMP-1 activity and collagen degradation possibly through the downregulation of MAPK/NF-κB/AP-1 signaling and SIRT1 activation.

Evaluation of collagenase, elastase and tyrosinase inhibitory activities of Cotinus coggygria Scop. through in vitro and in silico approaches

Cotinus coggygria Scop. (Anacardiaceae, syn: Rhus cotinus L.) is known as “boyacı sumağı, sarı boya, duman ağacı” in Turkish and “smoke tree” in English. It is commonly grown in Southern Europe and Anatolia. The leaves have been used due to its antiseptic, hemostatic, antipyretic, and wound healing effects as a 5% infusion in traditional medicine. It has also been reported to be used against skin disorders in Russia. Based on this information, the ethanol extracts prepared from the pedicels and leaves of C. coggygria were investigated for their elastase, collagenase, and tyrosinase inhibitory effects, which are enzymes related to anti-aging, using ELISA microtiter assays. Based on our results, the ethanol extracts prepared from the leaves and pedicels of C. coggygria had low elastase (28.16% ± 2.91 and 25.76% ± 1.71, respectively), moderate collagenase (47.78% ± 4.90 and 46.51% ± 3.15, respectively), and tyrosinase (57.94% ± 0.67 and 46.20% ± 0.92, respectively) inhibition at final concentration (666 µg/mL). The ethanol extract prepared from the pedicels of C. coggygria was subjected to bioactivity-guided fractionation, which led to isolation of methyl gallate, astragalin, isoquercetin, and hyperoside from the active fractions. In addition to the enzyme assays, in order to understand the inhibition mechanisms of the compounds inside the ligand-binding domains, the interactions were simulated and the key amino acids contributing to the hydrogen bonds and non-polar interactions with the ligands were reported.

Synthesis and characterization of new 4H-chromene-3-carboxylates ensuring potent elastase inhibition activity along with their molecular docking and chemoinformatics properties.

A new series of 4H-chromene-3-carboxylate derivatives were synthesized using multicomponent reaction of salicylaldehyde, ethyl acetoacetate and dimedone in ethanol with K3PO4 as a catalyst at 80°C. The structures of all newly synthesized compounds were confirmed by spectral techniques viz. IR, 1H NMR, 13C NMR, and LCMS analysis. The newly synthesized compounds 4a to 4j were screened against elastase enzyme. Interestingly, all these compounds found to be potent elastase inhibitors with much lower IC50 value. The compound 4b was found to be most potent elastase inhibitor (IC50=0.41±0.01µM) amongst the synthesized series against standard Oleanolic Acid (IC50 value=13.45±0.0µM). The Kinetics mechanism for compound 4b was analyzed by Lineweaver-Burk plots which revealed that compound inhibited elastase competitively by forming an enzyme-inhibitor complex. Along with this, all the synthesized compounds (4a - 4j) exhibits excellent DPPH free radical scavenging ability. The inhibition constant Ki for compound 4b was found to be 0.6µM. The computational study was comprehensible with the experimental results with good docking energy values (Kcal/mol). Therefore, these molecules can be considered as promising medicinal scaffolds for the treatment of skin-related maladies.

Antioxidant and Elastase Inhibitor from Black Soybean (Glycine max L.) and Its Compound (Daidzein)

Background: Skin aging occurs along with age. Genetic, nutrients, hormones, and sun exposure can affect directly and indirectly the structure of the skin. These things will induce an increase in free radicals which disrupts one of the most important enzymes in the extracellular matrix which is elastase enzyme. High enzyme synthesis will cause skin wrinkle. Free radicals can be inhibited by the presence of antioxidants. The plant contains natural phytochemical compounds which act as antioxidants and anti aging. Black soybeans are rich in daidzein compound, which protect against free radicals and prevent premature aging.Objective: This research evaluates the antioxidant and antiaging potential of Black Soybean (Glycine max L.) extract and its compound, daidzein.Methods: Analysis of antioxidants from black soybean extract and daidzein were carried out using ABTS scavenging activity assay. The antiaging assay was carried out through inhibition of elastase enzyme.Results: Black soybean extract had lower IC50 value of ABTS scavenging activity around 77.39±4.05 μg/ml better than daidzein with IC50 of 83.34±3.89 μg/ml. The results of elastase inhibition activity assay showed that daidzein compound has a lower IC50 value, 57.35±5.64 μg/ml compared to black soybean extract with IC50 value, 93.36±6.39 μg/ml.Conclusion: Black soybean extract had higher antioxidant. Daidzein had better elastase inhibition activity compared to black soybean extract.

Phytochemical analysis of olive flowers' hydroalcoholic extract and in vitro evaluation of tyrosinase, elastase and collagenase inhibition activity

Olea europaea L. is historically one of the most important trees of the Mediterranean countries. Increasing scientific interest regarding its fruits, leaves and olive oil has led to the elucidation of several phytochemical and biological characteristics. However, the phytochemical and biological studies regarding olive flowers remain limited. The aim of the present study was the phytochemical characterization of olive flowers' hydroalcoholic extract from Greek variety Lianolia, the effective isolation of the major secondary metabolites and evaluation of their inhibition activity against tyrosinase, elastase and collagenase. UPLC-HRMS/MS analysis was used to investigate the chemical composition of hydroalcoholic extract resulting in the identification of sixty-three secondary metabolites witch mainly belong to phenilethanoids, triterpenoids, flavonoids and secoiridoids. The orthogonial combination of Centrifugal Partition Chromatography and preparative HPLC in the same purification process led to the isolation of nine major compounds of the extract including two triterpenic acids, two flavonoid glycosides and five secoiridoid derivatives. From them, oleofloside A and oleofloside B are new natural products. Although, the hydroalcoholic extract and isolated secoiridoids exhibited weak or no inhibition activity towards tyrosinase and elastase, they exhibit remarkable anti-collagenase activity with 2΄-ethoxyoleuropein being the most active compound.

IN VITRO SCREENING OF ANTIOXIDANT AND ANTIAGING POTENTIAL OF CUCUMIS SATIVUS FRUIT EXTRACT

Objective: The present study was designed to screen the anti-aging and anti-wrinkle potential of Cucumis sativus fruit through in vitro estimation of antioxidant, anti-hyaluronidase, anti-elastase, anti-collagenase/anti-matrix metalloproteinase (MMP)-1, and anti-tyrosinase activity.&#x0D; Methods: Raw juice of cucumber was taken, filtered and fractionated with ethyl acetate and n-butanol. The obtained extracts were then evaluated for their antioxidant potential through 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay taking ascorbic acid as positive control and other enzymatic activities in reference to hyaluronidase inhibition, MMP-1/collagenase inhibition, and elastase inhibition taking catechin as reference standard whereas for tyrosinase inhibition the standard used was quercetin.&#x0D; Results: All the evaluations were performed in triplicates and results were noted down. It was observed that aqueous extract of C. sativus fruits showed a maximum DPPH radical scavenging activity (p&lt;0.0001), half-maximal inhibitory concentration (IC50) at a concentration of 122.67 μg/ml. The ethyl acetate fraction of C. sativus fruits exhibited maximum hyaluronidase (p&lt;0.0001), MMP-1/collagenase (p&lt;0.04), and tyrosinase (p&lt;0.04) inhibitory activity, IC50 at a concentration of 59.54, 45.79, and 24.46 μg/ml, respectively. The elastase (p&lt;0.0001) inhibitory activity by n-butanol fraction of C. sativus fruits extract was maximum, IC50 at a concentration of 52.76 μg/ml.&#x0D; Conclusion: A potent anti-aging and anti-wrinkle properties were well demonstrated by C. sativus, as depicted from the results obtained.

In vitro screening of elastase, collagenase, hyaluronidase, and tyrosinase inhibitory and antioxidant activities of 22 halophyte plant extracts for novel cosmeceuticals

BackgroundHalophyte plant (HPs), a salt-resistant flora, has been reported to provide several health benefits, but the knowledge of its cosmeceutical potential is still ambiguous. Here, 70% ethanol extracts of 22 HPs collected from along the coast of South Korea were investigated for their potentials of antioxidant, anti-aging, and whitening properties for use as materials in novel cosmeceuticals.MethodsAntioxidant activities were determined by DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical and hydrogen peroxide scavenging assays, and skin aging-related enzyme activities (anti-elastase, anti-collagenase, anti-hyaluronidase, and anti-tyrosinase) were evaluated by using the spectrophotometric method.ResultsAmong the 22 HPs, we found that Ischaemum antephoroides f. coreana and Atriplex gmelinii extracts presented the strongest scavenging effects against DPPH free radical and hydrogen peroxide, respectively. Our finding additionally suggested that Salicornia europaea extract might provide a major source of anti-elastase and anti-hyaluronidase; meanwhile, Rosa rugosa extract showed the highest anti-collagenase effect. Furthermore, the highest tyrosinase inhibitory activity was possessed by Spartina anglica extract.ConclusionThese findings may suggest that halophyte plants showing biological activities may be potent inhibitors of tyrosinase, elastase, collagenase, and hyaluronidase and could be useful for application in cosmeceuticals.

Investigation of Antioxidant and Elastase Inhibitory Activities of Geum urbanum Aerial Parts, Chemical Characterization of Extracts Guided by Chemical and Biological Assays

This study presents the bioguided chemical investigation of the 80% aqueous methanol extract of Geum urbanum aerial parts. Liquid–liquid partitioning of this extract in solvents of increasing polarity combined with biological screening showed that the ethyl acetate (EtOAc) soluble fraction was the most active part of the extract. This fraction was chemically profiled by a 13C nuclear magnetic resonance (NMR)-based dereplication method, resulting in the identification of 14 compounds. The dereplication process was followed by the purification of unknown and minor compounds of the EtOAc fraction. A new glycosylated phenol, namely, 3-(3,4-dihydroxyphenyl)propyl-α-l-rhamnopyranoside, together with 6 known compounds were isolated. Their structures were elucidated by spectroscopic methods including NMR and high-resolution electrospray ionization mass spectrometry. The antioxidant activity of fractions and isolated compounds were evaluated by 2,2,1-diphenyl-1-picrylhydrazyl and hydroxyl radical scavenging, and by cupric ion reducing antioxidant capacity assays. In parallel, their enzyme inhibitory property against human neutrophil elastase was assessed. Four subfractions, essentially containing polyphenols and triterpenes, exhibited a significant elastase inhibitory activity and an ellagitannin showed a very high radical scavenging activity.

Expression and purification of recombinant human serpin B1 yields novel molecules with altered protease inhibitory activities: Functional implications

Serpin B1 regulates the innate immune system by inhibiting serine and cysteine proteases that control programmed cell death and proliferation pathways. To provide recombinant human proteins for in vitro and in vivo studies we expressed and purified wild-type human serpin B1 and a C344A variant in the yeast S. cerevisiae. Both proteins expressed well and inhibited elastase and chymotrypsin. However, purification of wild-type serpin B1 in the absence of a reducing agent resulted in the specific loss of elastase - but not chymotrypsin - inhibition, concomitant with the formation of two higher molecular weight forms of the protein - a modified monomer and a dimer created via an intermolecular disulfide bond formed between C344 in respective serpin B1 monomers. In contrast to fully reduced serpin B1, both modified forms were good elastase substrates and catalytically cleaved at multiple adjacent sites within the reactive site loop. In contrast, purification of the C344A variant in the absence of a reducing agent yielded only one form of the protein which retained elastase and chymotrypsin inhibitory properties when purified. Furthermore, the elastase inhibitory activity of wild-type serpin B1, but not the C344A variant, was sensitive to oxidation. Thus, wild-type human serpin B1 should be formulated with a pharmaceutically acceptable reducing agent to protect C344 against post-translational oxidative modifications. Alternatively, the C344A variant of this protein may prove to be a suitable drug development candidate. These findings also suggest that inactivation of serpin B1 by oxidation may have a physiological role to play during inflammation.

Tutuilamides A-C: Vinyl-Chloride-Containing Cyclodepsipeptides from Marine Cyanobacteria with Potent Elastase Inhibitory Properties.

Marine cyanobacteria (blue-green algae) have been shown to possess an enormous capacity to produce structurally diverse natural products that exhibit a broad spectrum of potent biological activities, including cytotoxic, antifungal, antiparasitic, antiviral, and antibacterial activities. Using mass-spectrometry-guided fractionation together with molecular networking, cyanobacterial field collections from American Samoa and Palmyra Atoll yielded three new cyclic peptides, tutuilamides A-C. Their structures were established by spectroscopic techniques including 1D and 2D NMR, HR-MS, and chemical derivatization. Structure elucidation was facilitated by employing advanced NMR techniques including nonuniform sampling in combination with the 1,1-ADEQUATE experiment. These cyclic peptides are characterized by the presence of several unusual residues including 3-amino-6-hydroxy-2-piperidone and 2-amino-2-butenoic acid, together with a novel vinyl chloride-containing residue. Tutuilamides A-C show potent elastase inhibitory activity together with moderate potency in H-460 lung cancer cell cytotoxicity assays. The binding mode to elastase was analyzed by X-ray crystallography revealing a reversible binding mode similar to the natural product lyngbyastatin 7. The presence of an additional hydrogen bond with the amino acid backbone of the flexible side chain of tutuilamide A, compared to lyngbyastatin 7, facilitates its stabilization in the elastase binding pocket and possibly explains its enhanced inhibitory potency.

Inhibition of pancreatic elastase in silico and in vitro by Rubus rosifolius leaves extract and its constituents.

ABSTRACTObjective:Elastases are protease enzymes, which mainly hydrolyze proteins of the connective tissue, so they have a significant impact on human disease. Rubus rosifolius is one of the Rubus species found in Indonesian mountains, and it has potential as an elastase inhibitor. The objective of this research was to examine the in vitro elastase inhibitor activity of R. rosifolius leaves and to dock different ligands of its constituents against target protein of Porcine Pancreatic Elastase (PPE) receptor.Method:Dried leaves powder of R. rosifolius was extracted using Soxhlet apparatus with n-hexane, ethyl acetate, and methanol. The extract was evaporated, and in vitro elastase inhibitor activity was determined using PPE with the quercetin used as control positive. Selected nine constituents of R. rosifolius were evaluated on the docking behavior of elastase receptor using Protein–Ligand ANT System (PLANTS) computational software with PPE enzyme with Protein Data Bank (PDB) file 1BRU.Result:The methanol extract showed significantly inhibited elastase with IC50 186.13 μg/mL, but ethyl acetate extract showed weak activity, and n-hexane extract did not show any activity. Docking studies and binding free energy calculations and hydrogen bonding with some amino acids revealed that ellagic acid showed the least binding energy for the target enzyme.Conclusion:This research has opened new insights into understanding that constituents of R. rosifolius methanol extract are potential inhibitors against elastase, and suggested the active compound is ellagic acid.

적과 복숭아 추출물의 유용 성분 분석 및 추출조건별 기능성 소재 활용

Peaches are one of the most abundant cultivated fruits in Korea. There have been many studies on using it as functional materials; however, there has been few reports on immature peaches collected during the thinning out process. Herein we analyzed useful substances extracted under specific conditions from of thinned-out peaches and tested them for the suitability as cosmetic materials. NMR analysis revealed 3-O-cafffeoyl quinic acid as the major substance in the extracts. At PH-10, it showed positive response similar to ascorbic acid, used as a control, in antioxidant activity tests like DPPH and ABTS+ radical scavenging tests. The elastase and collagenase inhibitory activity tests showed minor wrinkle improvements for all the test groups. No antibacterial response was observed for five germ groups. These results show that immature peaches are applicable as functional materials because the major constituent of its extracts (chlorogenic acid) can act as an antioxidant at high concentrations. It could be of benefit to study immature peach extracts in more detail to potentially reveal further value-adding properties.

Assessment of fermented broken rice extracts for their potential as functional ingredients in cosmeceutical products

Rice-derived bioactive compounds are now widely used as active ingredients in health foods, pharmaceutical and cosmeceutical products and as food additives. Polyphenolic compounds, gamma oryzanol, and vitamin E are among the bioactive constituents in rice that have important antioxidant, anti-aging, and anti-pigmentation properties among others. In this present study, broken rice was subjected to solid-state fermentation for 20 days by using two types of fungi, Aspergillus niger and Rhizopus oligosporus. Cosmeceutical-related functionalities of the fermented broken rice extracts were tested by evaluating antioxidant activities as well as tyrosinase and elastase inhibition activities. Total phenolic content and total flavonoid content of fermented broken rice extracts were also measured. Fermentation caused a substantial increase in the total phenolic and flavonoid content of broken rice. Ferric-reducing antioxidant activity was the highest (18.47 mM FE(II)/g sample) in broken rice fermented with R. oligosporus on day 20. Fermentation of broken rice with the same fungus resulted in higher DPPH radical-scavenging antioxidant activity than A. niger, with the highest activity recorded was 94.17%. Broken rice fermented with R. oligosporus for 20 days had the highest tyrosinase and elastase inhibition activities with the value of 46% and 51.9%, respectively. The results demonstrate that the fermentation process increased the biological activities and the numbers of bioactive compounds for potential use in the cosmeceutical industry. This undervalued by-product of the rice milling industry can effectively be used for the production of high value products through fungal solid-state fermentation.

Anti-elastase activity of methanolic and ethyl acetate extract from Garcinia latissima Miq.

Elastase is a proteinase enzyme that can reduce elastin by dividing specific peptide bonds. Therefore, the inhibition of elastase activity in the dermis layer can be used to maintain skin elasticity. Materials that can inhibit elastase activity can be a cosmetic ingredient in dealing with skin aging. Increased desire to maintain healthy skin without chemicals, encourages the use of materials from natural sources such as plants from Indonesia, a country that has high biodiversity, including Garcinia latissima Miq. from the Clusiaceae family. The purpose of this study was to explore and sought an explanation as to why G. latissima Miq. inhibits elastase enzyme. Plant extraction was carried out using maceration successively methods. The inhibition activity of elastase enzyme was carried out by measuring the kinetic enzyme N-succ-(Ala)-3-Nitroanilide conversion to p-nitroaniline (substrate) spectrophotometry at 405 nm and using porcine pancreatic elastase (PPE) as an enzyme. The results showed that the G. latissima Miq. methanol extract and ethyl acetate extract were active as elastase enzyme inhibitors. G. latissima Miq. extract can maintain skin elasticity.

Bio-guided isolation of new phenolic compounds from Hippocrepis emerus flowers and investigation of their antioxidant, tyrosinase and elastase inhibitory activities

This study presents the bio-guided chemical investigation of a 80% methanol extract of Hippocrepis emerus flowers, a perennial non-climbing shrub. Liquid-liquid partitioning in solvents of increasing polarity combined to biological screening enabled to determine the EtOAc and n-BuOH soluble fractions as the most active parts of the extract. These fractions were chemically profiled by using a 13C NMR-based dereplication method, resulting in the identification of twenty-six compounds. The dereplication process was completed by purification of some unknown or minor compounds of the n-BuOH fraction. Three new glycosylated flavonoids, namely kaempferol-3-O-β-d-glucopyranosyl-7-O-β-d-glucopyranosyl-(1→3)-α-l-rhamnopyranoside (1), isorhamnetin-3-O-β-d-glucopyranosyl-7-O-β-d-glucopyranosyl-(1→3)-α-l-rhamnopyranoside (2) and quercetin-3-O-β-d-glucopyranosyl-7,4′-O-α-l-dirhamnopyranoside (3), together with twelve known compounds (4 – 15) were isolated. Their structures were elucidated by spectroscopic methods including NMR, HR-ESI-MS and UV. The antioxidant activity of fractions and isolated compounds were evaluated using DPPH and hydroxyl radical scavenging and CUPRAC assays. In parallel, their inhibitory properties against mushroom tyrosinase and human neutrophil elastase enzymes were assessed. Three quercetin glycosides exhibited a significant radical-scavenging activity and two flavonoids showed a moderate elastase inhibitory activity.

Production of α2,6-sialylated and non-fucosylated recombinant alpha-1-antitrypsin in CHO cells

Alpha-1-antitrypsin (A1AT) is an abundant serum inhibitor of serine proteases. A1AT deficiency is a common genetic disorder which is currently treated with augmentation therapies. These treatments involve weekly injections of patients with purified plasma-derived A1AT. Such therapies can be extremely expensive and rely on plasma donors. Hence, large-scale production of recombinant A1AT (rA1AT) could greatly benefit these patients, as it could decrease the cost of treatments, reduce biosafety concerns and ensure quantitative and qualitative controls of the protein. In this report, we sought to produce α2,6-sialylated rA1AT with our cumate-inducible stable CHO pool expression system. Our different CHO pools could reach volumetric productivities of 1,2 g/L. The human α2,6-sialyltransferase was stably expressed in these cells in order to mimic elevated α2,6-sialylation levels of native A1AT protein. Sialylation of the recombinant protein was stable over the duration of the fed-batch production phase and was higher in a pool where cells were sorted and enriched by FACS based on cell-surface α2,6-sialylation. Addition of ManNAc to the cell culture media during production enhanced both α2,3 and α2,6 A1AT sialylation levels whereas addition of 2F-peracetylfucose potently inhibited fucosylation of the protein. Finally, we demonstrated that rA1AT proteins exhibited human neutrophil elastase inhibitory activities similar to the commercial human plasma-derived A1AT.

Elastase, Hyaluronidase and Tyrosinase inhibitor activities antiaging of Curcuma mangga Val. extract and its fractions

Curcuma mangga Val. has bioactive compounds such as curcumin. Curcumin is a natural antioxidant and known as antiaging agent. Curcumin can stimulate the synthesis of collagen type I, inhibit melanogenesis and help to maintain correct skin hydration. This show C.mangga Val. has potential as antioxidant and antiaging agents. This research aims to find out the antiaging activities of C. mangga ethanolic extracts (CMEE) and its fractions. In this study, the antiaging activity was performed by measuring inhibition of elastase, hyaluronidase, and tyrosinase inhibitory activity assay. Ascorbic acid using as a control. In elastase inhibitory activity, ascorbic acid has the highest activity (15.53 µg/mL) but ethyl acetate fraction of C. Mangga (EACM) also has high activity (26.34 µg/mL) compared to other fractions and CMEE. In hyaluronidase inhibitory activity, ascorbic acid has the highest activity (27.67 µg/mL) but CMEE also has high activity (80.04 µg/mL) compared to other fractions, while in tyrosinase inhibitory activity EACM has the highest value (40.34 µg/mL) compared to ascorbic acid (65.99 µg/mL). In summary, C. mangga extracts have anti-hyaluronidase, while EACM also has good antiaging properties through anti-elastase and antityrosinase activity.