Investigation on the in vitro metabolism of bicyclol using liver microsomes, hepatocytes and human recombinant cytochrome P450 enzymes.

In vitro metabolism of bicyclol was studied using liver microsomes, hepatocytes and human recombinant cytochrome P450 enzymes. Liquid chromatography-benchtop orbitrap mass spectrometry technique was utilised to identify the metabolites.A total of 19 metabolites, including 5 new metabolites (M2, M3, M4, M5 and M16) were tentatively identified. Among these metabolites, M6&M8 (demethylenation), M9&M10 (demethylation) and M19 (glucuronidation) were the major metabolites.In glutathione (GSH)-supplemented liver microsomes, 5 new GSH conjugates were found and tentatively identified. The formation was assumed to be through demethylenation of methylenedioxyphenyl to form catechol derivatives, which further underwent oxidation to form ortho-quinone intermediates, reacting with GSH to form stable adducts.CYP3A4 and 2C19 were demonstrated to be the major enzymes responsible for the bioactivation of bicyclol.This study provided valuable information on the metabolic fate of bicyclol in liver microsomes and hepatocytes, and the bioactivation pathways were reported for the first time, which would be helpful for us to understand the potential drug-drug interactions and the possible side effect of this drug.

Identification and Strategies to Mitigate High Total Clearance of Benzylamine-Substituted Biphenyl Ring Systems.

For most oral small-molecule projects within drug discovery, the extent and duration of the effect are influenced by the total clearance of the compound; hence, designing compounds with low clearance remains a key focus to help enable sufficient protein target engagement. Comprehensive understanding and accurate prediction of animal clearance and pharmacokinetics provides confidence that the same can be observed for human. During a MERTK inhibitor lead optimization project, a series containing a biphenyl ring system with benzylamine meta-substitution on one phenyl and nitrogen inclusion as the meta atom on the other ring demonstrated multiple routes of compound elimination in rats. Here, we describe the identification of a structural pharmacophore involving two key interactions observed for both the MERTK program and an additional internal project. Four strategies to mitigate these clearance liabilities were identified and systematically investigated. We provide evidence that disruption of at least one of the interactions led to a significant reduction in CL that was subsequently predicted from rat hepatocytes using in vitro/in vivo extrapolation and the well-stirred scaling method. These tactics will likely be of general utility to the medicinal chemistry and DMPK community during compound optimization when similar issues are encountered for biphenyl benzylamines.

Fimasartan ameliorates renal ischemia reperfusion injury via modulation of oxidative stress, inflammatory and apoptotic cascades in a rat model.

Ischemia-reperfusion injury (IRI) can be defined as changes in the functions and structures of the tissues resulting from the restoration of blood after a period of ischemia. This study aimed to assess the potential protective effect of Fimasartan (angiotensin receptor antagonist) in the bilateral renal IRI in male rats through its potential effect on renal functions, modulation of the inflammatory cascade, oxidative stress, and apoptotic effect. The animals were equally assigned into four groups. The sham (negative control) group was exposed to surgical conditions without induction of IRI. The control group was exposed to ischemia by occluding the renal pedicles by clamps for 30 min, followed by restoration of blood for 2h. The vehicle-treated group received dimethyl sulfoxide (DMSO) by intraperitoneal injection (IP) 30 minutes before clamping. Fimasartan-treated group: rats pretreated with Fimasartan a dose of 3 mg/kg IP; this was half hour before occluding the renal pedicles. Animals were then exposed to 30 min ischemia (clamping the renal pedicles) followed by 2h reperfusion by releasing the clamps. Blood samples were collected to examine the levels of serum urea and creatinine. Renal tissue was used to measure the levels of cytokines (TNFα, IL-6) and total antioxidant capacity (TAC). Immunohistochemistry was used to assess the levels of Bax, caspase 3, and Bcl-2. Histopathological analyses were performed to detect the parenchymal injury. The present study shows that pretreatment with Fimasartan improves kidney function through its effects on oxidative stress, cytokines, and apoptotic markers.

LCZ696 Protects against Diabetic Cardiomyopathy-Induced Myocardial Inflammation, ER Stress, and Apoptosis through Inhibiting AGEs/NF-κB and PERK/CHOP Signaling Pathways.

The present study is designed to determine the effect of LCZ696 on DCM in rats and investigate the underlying mechanism involved. Diabetes was induced by feeding rats with a high-fat diet for six weeks following a single injection of STZ (30 mg/kg). Diabetic rats were divided into three groups (n = 10). LCZ696 and valsartan treatment was started two weeks after diabetic induction and continued for eight weeks. At the end of the treatment, serum and cardiac tissues were analyzed by RT-PCR, Western blot, and ELISA kits. LCZ696 and valsartan ameliorated DCM progression by inhibiting AGEs formation at activity levels; pro-apoptotic markers (BAX/Bcl2 ratio and caspase-3) in mRNA and protein expressions, the NF-κB at mRNA; and protein levels associated with the restoration of elevated proinflammatory cytokines such as the TNF-α, IL-6, and IL-1ß at the activity level. Furthermore, LCZ696 and valsartan contribute to restoring the induction of ER stress parameters (GRP78, PERK, eIF2a, ATF4, and CHOP) at mRNA and protein levels. LCZ696 and valsartan attenuated DCM by inhibiting the myocardial inflammation, ER stress, and apoptosis through AGEs/NF-κB and PERK/CHOP signaling cascades. Collectively, the present results reveal that LCZ696 had a more protective solid effect against DCM than valsartan.

Magnolol and Honokiol Inhibited the Function and Expression of BCRP with Mechanism Exploration.

Breast cancer resistance protein (BCRP), one of the ATP-binding cassette (ABC) transporters, was associated with the multidrug resistance (MDR) of chemotherapy. Magnolol (MN) and honokiol (HK) are major bioactive polyphenols of Magnolia officinalis. This study investigated the effects of MN and HK on the function and expression of BCRP for the purpose of developing BCRP inhibitor to overcome MDR. Cell lines including MDCKII-BCRP and MDCKII-WT were used for evaluating the function and expression of BCRP. The results showed that MN (100-12.5 µM) and HK (100-12.5 µM) significantly decreased the function of BCRP by 80~12% and 67~14%, respectively. In addition, MN and HK were verified as substrates of BCRP. Furthermore, MN and HK reduced the protein expression of BCRP, and inhibited the phosphorylation of epidermal growth factor receptor (EGFR) and phosphatidylinositol 3-kinase (PI3K). In conclusion, both MN and HK decreased the function and expression of BCRP via EGFR/PI3K signaling pathway. Therefore, both compounds were promising candidates for reversing the MDR of chemotherapy.

Antioxidant and tyrosinase inhibitory activities of traditional fermented Rosa from Dali Bai communities, Northwest Yunnan, China.

Traditional fermented Rosa (TFR) is a typical food and medical product among the Dali Bai people, and its popularity is growing. A few studies have looked into TFR's medicinal advantages, linked germplasm resources, traditional processing procedures, and functional food qualities. Our goal was to look into Rosa's traditional processing, examine the dominant strains in TFR, and prove how these strains affected antioxidant and tyrosinase inhibitory activities. We used a snowball selection strategy to pick 371 informants for a semi-structured interview, supplemented with direct observations and sample collection. A microbial strain was isolated and identified from a TFR sample collected in the field. We synthesized TFR in the lab using the traditional way. Both of 2, 2-diphenyl-1 picrylhydrazyl (DPPH) free radical scavenging and tyrosinase inhibitory properties of the fermented solution of Rosa 'Dianhong' have been tested in this study. Altogether 15 species belonging to the genus Rosa, which are utilized in herbal medicine and fermented foods. Rosa 'Dianhong' was the Bai community's principal species with considerable cultural value and consumption. Raw Rosa petals included 15 major flavonoids and phenols, which were identified as TFR's active components. TFR-1 was discovered to be the dominating microbial strain in TFR, increasing total phenolic and flavonoid content in the fermented solution of Rosa 'Dianhong' by 0.45 mg GAE/ml and 0.60 mg RE/ml, respectively, after 30 days. TFR-1 also exhibited promising activity in terms of DPPH free radical scavenging and tyrosinase inhibition. TFR showed potent antioxidant and free-radical scavenger properties and is beneficial in skincare and nutrition, according to the findings. TFR's medicinal and edible properties suggest that it could be used as a cosmetic or nutraceutical product.

Carbon dots derived from Beta vulgaris: evaluation of its potential as antioxidant and anticancer agent.

Carbon dots (CDs) endowed with outstanding physico-chemical characteristics expeditiously garnered tremendous popularity in the scientific community. CDs can be synthesized from a variety of natural resources and can replace metal semiconductor quantum dots in the range of applications such as bio-imaging, sensing and catalysis. Herein, CDs are green synthesized fromBeta vulgarisvia a single step hydrothermal approach (b-CDs). The synthesized carbon dots are characterized using UV-visible spectrophotometry, Fluorescence spectroscopy, High resolution transmission electron microscopy (HR-TEM), Fourier transform infrared spectroscopy (FT-IR), x-ray diffraction technique (XRD) and Raman spectroscopy. The b-CDs hence developed exhibited the signature 'excitation-dependent fluorescence emission' with its most intense emission in the green region. The quantum yield for the b-CDs obtained by this synthetic approach evinced an appreciable value of 11.6%. The antioxidant property of b-CDs are evaluated using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay to obtain a maximum scavenging activity of 94.5% at a concentration of 1000µg ml-1and its underlying mechanisms are illustrated. The blood compatibility of b-CDs are assessed using haemolysis assay and the cytotoxicity evaluated using MTT assay shows significant cell growth-inhibition against the human breast cancer (MCF-7) and hepatocellular carcinoma (HepG2) cell lines. This succinct study demonstrates the inherent therapeutic potential of biocompatible carbon dots.

Application of the Life Cycle Management of Analytical methods concept to a HPTLC-DPPH assay method for acteoside content in industrial extracts of Plantago lanceolata L.

Analytical methods used for quality control of plants and plant extracts are based on the identification and quantification of chemical markers to manage batch reproducibility and efficacy. The aim of this work was to assess the performance of a High Performance Thin Layer Chromatography (HPTLC) method developed for quality control of industrial dry extracts of ribwort plantain (P. lanceolata L.), using 2,2-diphenyl 1-picrylhydrazyle (DPPH) effect directed chemical reaction for antioxidant activity of acteoside, a phenylethanoid glycoside commonly used as a marker for P. lanceolata L., and to demonstrate the applicability of the Life Cycle Management of Analytical Methods concept to quantitative HPTLC-DPPH methods. The first step was the determination of the Analytical Target Profile (ATP) and Target Measurement Uncertainty (TMU), taking into account the quality control requirements for such extracts and the detection method applicable range. Once the desired range was established, an evaluation of the calibration function was conducted using several calibration models. Due to the lack of reference samples, spiked samples were used to evaluate the accuracy of the method by means of Total Analytical Error (TAE) determination, using prediction intervals calculation for the selected calibration functions. Measurement Uncertainty (MU) was also estimated, allowing the final choice of the calibration function to be used for quality control, giving the most fit for purpose performance level in accordance with the product specifications. As Life Cycle Management of the method also includes its routine use, the Measurement Uncertainty was checked on spiked and unspiked extract samples with different dilution levels, in order to verify the accordance of results between spiked and unspiked samples and to prepare a replication strategy to be applied during the routine use of the method.

Radical scavenging activity and metabolomic profiling study of ylang-ylang essential oils based on high-performance thin-layer chromatography and multivariate statistical analysis.

Ylang-ylang (YY) essential oil (EO) is distilled from the fresh-mature flowers of the Annonaceae family tropical tree Cananga odorata [Lam.] Hook. f. & Thomson, and is widely used in perfume and cosmetic industries for its fragrant character. Herein, two different metabolomic profiles obtained using high-performance thin-layer chromatography (HPTLC), applying different stains, namely 2,2-diphenyl-1-picrylhydrazyl (DPPH·) and p-anisaldehyde, were used for discrimination of 52 YY samples across geographical origins and distillation grades. The first profile is developed using the DPPH· stain based on the radical scavenging activity (RSA) of YY EOs. Results of the HPTLC-DPPH· assay confirmed that RSA of YY EOs is in proportion to the length of distillation times. Major components contributing to the RSA of YY EOs were tentatively identified as germacrene D and α-farnesene, eugenol and linalool, by gas chromatography-mass spectrometry (GC-MS) and GC-flame ionisation detector (GC-FID). The second profile was developed using the general-purpose p-anisaldehyde stain based on the general chemical composition of YY EOs. Untargeted metabolomic discrimination of YY EOs from different geographical origins was performed based on the HPTLC-p-anisaldehyde profiles, followed by principal component analysis (PCA). A discrimination and prediction model for identification of YY distillation grade was developed using PCA and partial least squares regression (PLS) based on binned HPTLC-ultraviolet (254 nm) profiles, which was successfully applied to distillation grade determination of blended YY Complete EOs.

Potential Antioxidant and Anti-Inflammatory Function of Gynura procumbens Polyphenols Ligand.

The antioxidant and anti-inflammatory potentials of polyphenols contained in Gynura procumbens (GP) extract were systematically analyzed. Polyphenols in GP were analyzed for nine peaks using high-performance liquid chromatography (HPLC) combined with mass spectrometry (MS), and quantitatively determined through each standard. A total of nine polyphenolic compounds were identified in the samples and their MS data were tabulated. To determine the potential of bioactive ingredients targeting DPPH and COX-2, we analyzed them by ultrafiltration combined with LC. The results identified the major compounds exhibiting binding affinity for DPPH and COX-2. Caffeic acid, kynurenic acid, and chlorogenic acid showed excellent binding affinity to DPPH and COX-2, suggesting that they can be considered as major active compounds. Additionally, the anti-inflammatory effect of GP was confirmed in vitro. This study will not only be used to provide basic data for the application of GP to the food and pharmaceutical industries, but will also provide information on effective screening methods for other medicinal plants.

Pharmacokinetics-Driven Evaluation of the Antioxidant Activity of Curcuminoids and Their Major Reduced Metabolites-A Medicinal Chemistry Approach.

Curcuminoids are the main bioactive components of the well-known Asian spice and traditional medicine turmeric. Curcuminoids have poor chemical stability and bioavailability; in vivo they are rapidly metabolized to a set of bioreduced derivatives and/or glucuronide and sulfate conjugates. The reduced curcuminoid metabolites were also reported to exert various bioactivities in vitro and in vivo. In this work, we aimed to perform a comparative evaluation of curcuminoids and their hydrogenated metabolites from a medicinal chemistry point of view, by determining a set of key pharmacokinetic parameters and evaluating antioxidant potential in relation to such properties.Reduced metabolites were prepared from curcumin and demethoxycurcumin through continuous-flow hydrogenation. As selected pharmacokinetic parameters, kinetic solubility, chemical stability, metabolic stability in human liver microsomes, and parallel artificial membrane permeability assay (PAMPA)-based gastrointestinal and blood-brain barrier permeability were determined. Experimentally determined logP for hydrocurcumins in octanol-water and toluene-water systems provided valuable data on the tendency for intramolecular hydrogen bonding by these compounds. Drug likeness of the compounds were further evaluated by a in silico calculations. Antioxidant properties in diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and oxygen radical absorbance capacity (ORAC) assays were comparatively evaluated through the determination of ligand lipophilic efficiency (LLE). Our results showed dramatically increased water solubility and chemical stability for the reduced metabolites as compared to their corresponding parent compound. Hexahydrocurcumin was found the best candidate for drug development based on a complex pharmacokinetical comparison and high LLE values for its antioxidant properties. Development of tetrahydrocurcumin and tetrahydro-demethoxycurcumin would be limited by their very poor metabolic stability, therefore such an effort would rely on formulations bypassing first-pass metabolism.

Phytochemical, antibacterial, antioxidant and cytoxicity investigation of Tarenna grandiflora.

The phytochemical investigation of Tarenna grandiflora led to the isolation of 18 known compounds of which were four flavones, three anthraquinones, one phenyl propanoic derivative, five triterpenoids, four steroids and a mixture of glucose. Luteolin (1) and soranjidiol (6) were allylated and/or prenylated to give four new semisynthesized derivatives which were fully characterized as 7,3',4'-O-triallylluteolin (1a), luteolin-7,3',4'-O-triprenyls (1b), luteolin-5,7,3',4'-O-tetraprenyls (1c) and 6-O-allylsoranjidiol (6a). Their structures were established using spectroscopic analysis including 1D, 2D NMR and MS data. The cytotoxic, antioxidant and antimicrobial activities of extracts, fractions, isolated compounds and semi-synthesized derivatives were evaluated. The petroleum ether and EtOAc extracts exhibited good cytotoxic potency on KB-3-1 cell line with IC50 of >0.1 and 0.025 mg/mL respectively, while compounds 1b and 11 were the most active (IC50 > 0.0001 M). Compounds 1 and 3 showed the best antioxidant activities (45.5 and 55.8 µM); while compounds 9 and 12 showed the best antibacterial activities with MICs values ranges from 8.55 to 132 µM.

Talinum paniculatum (Jacq.) Gaertn. leaves - source of nutrients, antioxidant and antibacterial potentials.

BACKGROUND: The diet of most of the population is limited to a reduced number of plants, even in areas that have a varied and extensive diversity, such as Brazil. Unconventional Food Plants (UFPs) are plants considered exotic, native, and wild that grow naturally and can be used as food. Among these is Talinum paniculatum (Jacq.) Gaertn., which is widespread throughout Brazil and can be a potential source of nutrients. Due to the potential of utilization of UFPs in human food and the lack of studies regarding the composition of T. paniculatum, this study aimed to assess the nutritional value of T. paniculatum leaves, their antioxidant capacity, and their antimicrobial activity for possible use in food. METHODS: The characterization of the leaves of T. paniculatum was carried out through analyses of proximal composition, color, ascorbic acid, mineral profile, and antinutritional factors showing the presence of condensed and hydrolysable tannins and nitrates in low concentrations. Solvents of water, ethanol, ethanol/water, methanol, methanol/water, methanol/acetic acid and acetone/water/acetic acid were used to evaluate the extraction yield of phenolic compounds, antioxidant capacity, and antibacterial activity of the extracts. RESULTS: High contents of protein (18.61 g 100 g-1), insoluble dietary fiber (34.75 g 100 g-1), ascorbic acid (81.03 mg 100 g-1), magnesium, potassium, and calcium (649.600, 411.520 and 228.117 mg 100 g-1, respectively) were observed. Extraction using the mixture of solvents of methanol/acetic acid showed the highest yield of phenolic compounds (432.73 mg EAG 100 g-1) and antioxidant capacity using the DPPH assay (3144.92 mg 100 g-1). Bacillus cereus growth was inhibited by the T. paniculatum extracts. CONCLUSIONS: T. paniculatum leaves are a source of nutrients and their extracts have antioxidant and antibacterial potentials which can be used as supplements in food to improve one's health.

Optimization of ultrasound-assisted extraction of polyphenols from globe artichoke (Cynara scolymus L.) bracts residues using response surface methodology.

BACKGROUND: The globe artichoke (Cynara scolymus L.) is a rich source of phenolic compounds which may be extracted by ultrasound technology and used as a medicinal alternative. The objective of this work was to determine the radiation amplitude (%), ethanol concentration (%), and time extraction (min) required to guarantee an elevated content of polyphenol compounds. METHODS: The optimal extraction conditions were assessed through the Box-Wilson design and by applying Composite Face Centered (CCFC) and total phenolic compounds (TPC) as the response variables. RESULTS: A quadratic model was adequate, with R2 = 0.993. The optimal conditions were a radiation amplitude of 97%, an ethanol concentration of 53%, and an extraction time of 9.7 min. The optimized extract of artichoke bracts (Cynara scolymus L.) showed a TPC of 25.13 (±0.030) mg GAE/g, an antioxidant activity DPPH of 39.79 (±0.014) mmol Trolox equivalents (TE), and an antioxidant capacity TEAC of 33.98 (±0.03) mmol Trolox equivalents. CONCLUSIONS: The results showed values closely related to the expected values, indicating that the models were well-developed.

Antibacterial, antifungal and antioxidant activities of whole plant chemical constituents of Rumex abyssinicus.

BACKGROUND: Antibiotic resistance has contributed to the burden of infectious diseases both in the hospital and community setting, and represents a great threat to public health. Previous studies have revealed the role of reactive oxygen species as intermediate mediators of tissue damage, following antibiotherapies, indicating the need of associating antioxidants to these treatments. Therefore, the present work was designed to study the antibacterial, antifungal and antioxidant activities of extracts and compounds from Rumex abyssinicus Jacq. (Polygonaceae), as well as to investigate the antibacterial mechanisms of action of the most effective agents. METHODS: The plant extracts were prepared by maceration in organic solvents followed by column chromatography of the EtOAc fraction and purification of different fractions which led to the isolation and characterization of pure compounds. The antimicrobial activities of the extracts/compounds and their combinations with ciprofloxacin and fluconazole were evaluated using the broth microdilution method by determining the minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC). The effects of the extracts on the bacterial cell membrane and microbial respiratory chain dehydrogenase enzyme activity were determined by spectrophotometric methods. Antioxidant activity was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and gallic acid equivalent antioxidant capacity (GAEAC) assays. RESULTS: Chrysophanol (1), physcion (2), Ergosta-6,22-diene-3,5,8-triol (3), emodin (4), 6-hydroxyemodin (citreorosein) (5), chrysophanein (6) and physcionin (7) were isolated from EtOAc fraction of R. abyssinicus and displayed different degrees of antimicrobial activities (MIC = 8-256 µg/mL). The MeOH extract and compounds 2 and 4 exhibited synergistic effects with ciprofloxacin and fluconazole. Compounds 1, 2 and the combined mixture of 6 + 7 displayed the highest antioxidant activity (GAEAC = 83.38-106.03 µg/mL). CONCLUSION: R. abyssinicus is a potential source of antibacterial, antifungal and antioxidant agents. The antibacterial mechanisms of action of the MeOH extract and compound 2 are due to disruption of the cytoplasmic membrane and inhibition of the microbial respiratory chain dehydrogenase enzyme activity. To the best of our knowledge, this is the first report of test samples and ciprofloxacin / fluconazole association against MDR strains. The observed activity of the isolated compounds against bacteria and fungi including MDR strains deserves further exploration.

Biological evaluation of Safrole oil and Safrole oil Nanoemulgel as antioxidant, antidiabetic, antibacterial, antifungal and anticancer.

BACKGROUND: Safrole is a natural compound extracted from various plants, and has shown various biological activities. The current study aimed to investigate the antioxidant, antidiabetic, antimicrobial, and anticancer activity of safrole oil and to study the influence of safrole nanoemulgel on these activities. METHODS: The antioxidant and antidiabetic in-vitro assays were conducted using standard biomedical methods. The safrole oil nanoemulgel was developed using a self-emulsifying technique. Then the antimicrobial activity of the safrole oil and safrole nanoemulgel were performed on different microbial species, and cytotoxicity was determined against Hep3B cancer cell lines using the MTS assay. RESULTS: Safrole oil showed moderate antioxidant activity compared with standard Trolox, with IC50 value 50.28 ± 0.44 and 1.55 ± 0.32 µg/ml, respectively. Moreover, it had potent α-amylase inhibitory activity (IC50 11.36 ± 0.67 µg/ml) compared with Acarbose (IC50 value 5.88 ± 0.63). The safrole nanoemulgel had pseudo-plastic behaviour, droplet sizes below 200 nm, a polydispersity index (PDI) below 0.3, and a zeta potential of less than - 30 mV. Safrole oil has potential antimicrobial and anticancer activities, and these activities were improved with safrole nanoemulgel. CONCLUSION: The safrole oil may be applied for the prevention and treatment of oxidative stress, diabetes, different microbial species and cancer, and these activities could be improved by nano-carriers.

Post-translational modifications and antioxidant properties of different therapeutic human serum albumins.

Human serum albumin (HSA) is widely used for the treatment of diverse clinical conditions to restore plasma volume, manage burns and treat hypoproteinemia.Although the HSA preparations should ideally preserve its functionality, the structural integrity and antioxidant properties of HSA may be compromised as a result of the manufacturing process. The present study examined seven commercially available HSA preparations for clinical use to investigate their post-translational modifications (PTMs) and antioxidant activity, including DPPH radical-scavenging, peroxyl radical antioxidant and metal binding activities, by means of mass spectrometry and Ellman's assay. The results confirmed that most of the PTMs of HSA and especially the oxidation of the free thiol residue varied between the different commercial albumins and the percentage of these PTMs were higher than those of physiological HSA. Moreover, HSA-DA isoform was increased at the end of the stability time and new oxidative modifications occurred in these samples. In conclusion, the bioprocesses for production of commercial albumins are responsible of their wide heterogeneity, being the ethanol fractionation and their storage conditions the more critical phases. Nonetheless, the Kedrion albumin shows a high content of free thiol and a lower concentration of PTMs than other commercial albumins.

RM12 similar to substance P from tachykinin of freshwater murrel Channa striatus influence intracellular ROS in vitro fish erythrocytes and developmental toxicity and antioxidant enzymes in vivo zebrafish embryo.

In this study, substance P, an antioxidant peptide of tachykinin, was identified using bioinformatics tools from the earlier established muscle transcriptome of a freshwater murrel Channa striatus and the peptide was named RM12. The antioxidant properties of RM12 were screened using various colorimetric assays. The toxicity of RM12 was experimented using fish erythrocytes, and it is observed that the maximum concentration (320 µM) of RM12 was found to have 15 or 20% of hemolytic activity; however, it was not significant with other tested concentrations (10, 20, 40, 80, and 160 µM). Further, the in vivo antioxidant properties of RM12 were experimented on zebrafish embryo, the intracellular ROS level was estimated by 5 mM H2O2 stress in the zebrafish embryo, and inhibition of apoptosis was evaluated. The antioxidant enzymes were extracted from the H2O2-stressed zebrafish embryo, and the intracellular ROS was eliminated due to RM12. Collectively, the experiment showed that the substance P from the freshwater murrel C. striatus possessed potent antioxidant properties; thus, it can further be focused to develop it as antioxidant molecule in aquaculture organisms.

Discovery of G Protein-Biased Ligands against 5-HT7R.

There has been significant attention concerning the biased agonism of G protein-coupled receptors (GPCRs), and it has resulted in various pharmacological benefits. 5-HT7R belongs to a GPCR, and it is a promising pharmaceutical target for the treatment of neurodevelopmental and neuropsychiatric disorders. Based on our previous research, we synthesized a series of 6-chloro-2'-methoxy biphenyl derivatives 1, 2, and 3 with a variety of amine scaffolds. These compounds were evaluated for their binding affinities to 5-HTR subtypes and their functional selectivity toward the Gs protein and the ß-arrestin signaling pathways of 5-HT7R. Among them, 2-(6-chloro-2'-methoxy-[1,1'-biphenyl]-3-yl)-N-ethylethan-1-amine, 2b, was found to be a G-protein-biased ligand of 5-HT7R. In an in vivo study with Shank3 transgenic mice, the self-grooming behavior test was performed with 2b, which increased the duration of self-grooming. The experiments further suggested that 5-HT7R is associated with autism spectrum disorders (ASDs) and could be a therapeutic target for the treatment of stereotypy in ASDs.

Novel Dual-Target µ-Opioid Receptor and Dopamine D3 Receptor Ligands as Potential Nonaddictive Pharmacotherapeutics for Pain Management.

The need for safer pain-management therapies with decreased abuse liability inspired a novel drug design that retains µ-opioid receptor (MOR)-mediated analgesia, while minimizing addictive liability. We recently demonstrated that targeting the dopamine D3 receptor (D3R) with highly selective antagonists/partial agonists can reduce opioid self-administration and reinstatement to drug seeking in rodent models without diminishing antinociceptive effects. The identification of the D3R as a target for the treatment of opioid use disorders prompted the idea of generating a class of ligands presenting bitopic or bivalent structures, allowing the dual-target binding of the MOR and D3R. Structure-activity relationship studies using computationally aided drug design and in vitro binding assays led to the identification of potent dual-target leads (23, 28, and 40), based on different structural templates and scaffolds, with moderate (sub-micromolar) to high (low nanomolar/sub-nanomolar) binding affinities. Bioluminescence resonance energy transfer-based functional studies revealed MOR agonist-D3R antagonist/partial agonist efficacies that suggest potential for maintaining analgesia with reduced opioid-abuse liability.