Half-maximal Effective Concentration Research Articles

The number of test organisms might influence the toxicity evaluation of hydrophobic micropollutants

Evaluating the toxicity of micropollutants forms the basis for understanding their potential risks to the ecosystem and/or human health. To accurately evaluate the toxicity of micropollutants in toxicity tests, many factors have been carefully considered, while the impact of the number of test organisms on toxicity results has rarely been taken into account. In this study, the role of the organism number on the developmental toxicity of five micropollutants was investigated using embryos of the marine polychaete Platynereis dumerilii. The toxicity of hydrophobic micropollutants was found to decrease significantly with increasing the number of embryos used in the test. A quantitative model was developed to better describe how the number of embryos affected developmental toxicity. The model showed a satisfactory fit to the raw data in all scenarios tested. The intrinsic half-maximal effective concentration EC50,int was then determined using the model. For a given compound, the EC50,int was a stable parameter that did not depend on the number of test embryos and thus provided an indication of the intrinsic toxicity of the compounds tested. Compared with the EC50 values determined with the commonly used embryo number (around 120), the EC50,int values of all tested hydrophobic micropollutants were lower. The more hydrophobic the compounds tested, the more pronounced the reduction in toxicity. This suggested that hydrophobic micropollutants could be more toxic than reported in the literature. Some suggestions were also made to eliminate the effect of the number of organisms used in the toxicity evaluation.

Using Folsomia candida (Collembola) for the ecological assessment of sediment samples from three rivers from the QwaQwa region, South Africa

AbstractSince the establishment of a free economic zone and a simultaneous increase in the population of the QwaQwa region, aquatic systems of the area were expected to undergo an increase in metal contamination. The aims of this study were to assess the quality of sediments from the Metsi-Matsho, Namahadi and Elands Rivers of the QwaQwa region, and to investigate their impact on the survival and reproduction of the collembolan Folsomia candida. Due to the lack of freshwater sediment quality guidelines in South Africa, metal contents of the sediments were analysed and critically interpreted using the national action list for the screening of dredged sediment proposed for marine disposal, and international sediment quality guidelines (SQGs). In the laboratory, F. candida was exposed to 0, 50, 75 and 100% of the sediment samples for 28 days at 20 °C. Sediments were variably contaminated with Cr, with the Namahadi River middle site having the highest concentration (569 mg.kg−1, dry weight) followed by the lower site on the Elands River (529 mg.kg−1, dry weight), both above the recommended prohibition level of 500 mg Cr.kg−1. According to the consensus-based SQGs, only the concentrations of Cr and Ni (upper reach of Metsi-Matsho River) were higher than probable effect concentration, denoting that harmful effects of those metals are likely to occur. Of the two endpoints assessed, reproduction was the most sensitive with the upper reach of the Metsi-Matsho River (half maximal effective concentration, EC50 = 24% sediment) being the most deleterious site to the reproduction of F. candida. Ecotoxicological bioassays using F. candida could be suitable complements to chemical analysis in the assessment of the ecological risks of freshwater sediments as this collembolan species is sensitive to sediments with variable ranges of metal contamination.

Anti-Leishmanial Effects of a Novel Biocompatible Non-Invasive Nanofibers Containing Royal Jelly and Propolis against Iranian Strain of Leishmania major (MRHO/IR/75/ER): an In-Vitro Study

Background: Current medications especially the pentavalent antimonial compounds have been used as the first line treatment of cutaneous leishmaniasis (CL), but they have limitations due to serious side effects such as drug resistance, cardio and nephrotoxicity, and high costs. Hence, the demand to find more usable drugs is evident. Synthesis and devel­opment of natural, effective, biocompatible, and harmless compounds against Leishmania major is the principal priority of this study.
 Methods: By electrospinning method, a new type of nanofiber were synthesized from royal jelly and propolis with dif­ferent ratios. Nanofibers were characterized by Scanning Electron Microscope (SEM), Transmission Electron Micros­copy (TEM), Thermogravimetric Analysis (TGA), Contact angle, and Fourier-transform infrared spectroscopy (FTIR). The Half-maximal inhibitory concentration (IC50), Half-maximal effective concentration (EC50) and the 50% cytotoxic concentration (CC50) for different concentrations of nanofibers were determined using quantitative calorimetric meth­ods. Inductively coupled plasma-optical emission spectrometry (ICP-OES) and flow cytometry were performed as com­plementary tests.
 Results: The results showed that the proposed formulas provide a new achievement that, despite the significant killing activity on L. major, has negligible cytotoxicity on the host cells. Royal jelly nanofibers have significantly shown the best 72 hours results (IC50= 35 μg/ml and EC50=16.4 μg/ml) and the least cytotoxicity.
 Conclusion: This study presents a great challenge to introduce a new low-cost treatment method for CL, accelerate wound healing, and reduce scarring with minimal side effects and biocompatible materials. Royal jelly and propolis nanofibers significantly inhibit the growth of L. major in-vitro.

Sensory and molecular insights into the bitterness of Wuyi rock tea

Wuyi Rock Tea (WRT) is widely appreciated for its unique rocky flavor and floral essence. However, the characterization of taste attributes, particularly bitterness, in this premium tea remains largely underexplored. This study aims to assess the perception of bitterness in WRT through an integrated approach, including sensory evaluation, phenolic compound analysis, and cell-based calcium imaging assay. Sensory evaluation revealed distinct bitterness intensities among nine selected typical WRT samples. Utilizing UHPLC-Q-Orbitrap-MS, 54 phenolics were tentatively identified, comprising of 24 flavanols, 18 flavonols, and 12 phenolic acids. The cell-based calcium imaging assay revealed diverse intracellular calcium responses and half-maximum effective concentration (EC50) values across the tested samples. Notably, correlation analysis unveiled a strong negative correlation between sensory scores and EC50 values. Furthermore, 11 individual phenolic compounds, such as epigallocatechin gallate and epicatechin gallate, demonstrated significant correlation with sensory scores and bitter taste receptor activation, suggesting their potential role as key contributors shaping the bitter taste perception of WRT. This study provides comprehensive insights into the nuanced bitterness of WRT, bridging sensory and molecular perspectives to enhance our understanding of the taste profile of this premium tea.

Стрессорное воздействие биоцидных метаболитов актинобактерии Streptomyces geldanamycininus Z374 на цианобактерии Microcystis aeruginosa

Over the past decades, as a result of anthropogenic pollution and global warming, the frequency, intensity and geographical distribution of the mass development of microalgae and cyanobacteria in water bodies have significantly increased. The Microcystis species are responsible for most cases of the cyanobacterial blooms in freshwater bodies worldwide. Microcystis aeruginosa is the most commonly observed of them. In this work, we assessed the stress effect of isolated from actinobacteria Streptomyces geldanamycininus cyanocidal crude biocide Z374 (CB Z374) on the cyanobacteria M. aeruginosa. The results showed that CB Z374 significantly inhibited M. aeruginosa biomass. The CB Z374 half maximal effective concentration (EC50) for M. aeruginosa after 96 hours of treatment was 5 mg/L. The growth of M. aeruginosa with CB Z374 is characterized by an increase in the duration of the lag-phase, a decrease in the specific growth rate and biomass yield compared to the control. CB Z374 caused oxidative stress in cyanobacteria cells, determined by the elevated levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA) up to 1.6 times, as well as an increased permeability of M. aeruginosa cell membranes by 1.5 times. The levels of superoxide dismutase (SOD) and catalase (CAT) activities and reduced glutathione (GSH) content raised in response to oxidative stress. M. aeruginosa growth inhibition was accompanied by a photosynthesis modification: the photosynthetic pigments phycobiliproteins and carotenoids content increases by 2.3 and 1.6 times respectively and the chlorophyll a content decreases by 1.2 times. CB Z374 inhibited M. aeruginosa photosynthesis, which was expressed in a decline in the maximum quantum efficiency of photosystem II – Fv/Fm by 10% and a reduction in the electron transfer rate (ETR) by 18%. We assume that oxidative stress and, as a result, cell membranes damage and photosynthesis inhibition are the main mechanisms of the cyanocidal action of CB Z374 on M. aeruginosa.

Abstract 1032: Development of a novel companion diagnostic immunohistochemistry antibody for claudin 18.2-targeted therapies

Abstract Background: CLDN18.2, the isoform 2 of CLDN18, is physiologically confined to gastric mucosa tight junctions, the epitopes of which would be exposed on the cancer cell surface upon malignant transformation and is highly expressed in a significant proportion of cancers, e.g., gastric and pancreatic adenocarcinomas, which makes it a promising drug target for cancer therapy. Currently, multiple drug modalities, such as monoclonal antibodies, ADCs, and CARTs targeting CLDN18.2 are being evaluated in clinical trials. A substantial correlation exists between the expression of CLDN18.2 and the efficacy of the these therapies. However, to date, no companion diagnostic (CDx) antibodies that are specific to CLDN18.2 have been approved. Therefore, it is important to promptly develop an anti-CLDN18.2 CDx antibody with a high specificity, sensitivity, and suitability for immunohistochemistry (IHC). In this study, we present the discovery and validation of a novel, highly sensitive IHC antibody that selectively identifies CLDN18.2. Methods: 3 CLDN18.2 specific peptide fragments from extracellular domain of human CLDN18.2 were conjugated with KLH. Antibodies were discovered by immunizing BALB/c and SJL mice with these peptides-KLH conjugates. Screening and selection process was relied on positive ELISA results against the immunogen, as well as FACS and Acumen Explorer Microplate staining of CLDN18.2-experssing cells. Hit antibodies were evaluated for IHC on formalin-fixed paraffin-embedded CHOK1 cells expressing full-length CLDN18.2 or CLDN18.1. The antibodies were further validated on human gastric cancer tissue samples to establish the analytical sensitivity and specificity of the IHC assay. Results: The mAb clone 43F11 exhibited binding to a fusion polypeptide comprising the first extracellular loop of human CLDN18.2 (aa32-53) at a half maximal effective concentration (EC50) of 0.212 nM. 43F11 showed positive cell surface IHC staining on CLDN18.2-experssing cells after fixation but demonstrated no staining on CLDN18.1-experssing cells. The IHC test method was established by utilizing clinical tumor tissues and PDX samples that had predetermined expression levels of CLDN18.2. The 43F11 antibody accurately identified the expression level of CLDN18.2 in the IHC assay, with a minimum concentration of 0.25 μg/mL. When compared to the commercial CLDN18.2 IHC antibody EPR19202, 43F11 demonstrated greater sensitivity, allowing for positive staining on cancer tissues that have significantly lower levels of CLDN18.2 expression. Conclusion: 43F11 exhibits specific recognition of human CLDN18.2 in IHC assays, while showing no reactivity towards CLDN18.1. It demonstrated superior sensitivity compared to the benchmark antibody. These data suggest that 43F11 has the capacity to serve as an effective diagnostic tool for identifying patients who positively express CLDN18.2. Citation Format: Jishun Chen, Suya Bai, Yu Bai, Zaoshun Hu, Peng Chen, Jay Mei, Bo Shan, Bing Hou. Development of a novel companion diagnostic immunohistochemistry antibody for claudin 18.2-targeted therapies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1032.

Population Pharmacodynamic Dose-Response Analysis of Serum Potassium Following Dosing with Sodium Zirconium Cyclosilicate.

Sodium zirconiumcyclosilicate (SZC) is an approved oral treatment for hyperkalemia that selectively binds potassium (K+) in the gastrointestinal tract and removes K+ from the body through increased fecal excretion. Here, we describe the population pharmacodynamic (PopPD) response of serum K+ concentration in patients with hyperkalemia who are treated with SZC, estimate the impact of patients' intrinsic and extrinsic factors, and compare predicted serum K+ responses between 5 g alternate daily (QOD) and 2.5 g once daily (QD) maintenance doses. PopPD analysis was based on pooled data from seven phase II and III clinical trials for SZC. A semi-mechanistic longitudinal mixed-effects (base) model was used to characterize serum K+ concentration after SZC dosing. Indirect-response, virtual pharmacokinetics-pharmacodynamics (PK-PD) modeling was used to mimic the drug exposure compartment. Full covariate modeling was used to assess covariate impact on the half-maximal effective concentration of drug (EC50), placebo response, and Kout. Models were evaluated using goodness-of-fit plots, relative standard errors, and visual predictive checks, and data were stratified to optimize model performance across subgroups. Covariate effects were evaluated based on the magnitude of change in serum K+ between baseline and end of correction phase dosing (48 h, SZC 10 g three times a day) and maintenance phase dosing (28days, SZC 10 g QD) using a reference subject. The analysis data set included 2369 patients and 25,764 serum K+ observations. The mean (standard deviation) patient age was 66.0 (12)years, 61% were male, 68% were White, 34% had congestive heart failure, and 62% had diabetes. Mean (standard deviation) serum K+ at baseline was 5.49 (0.43)mmol/L. Both the base and full covariance models adequately described observed data. In the final model, there was a sigmoid exposure response on Kin, with EC50 of 32.8 g and a Hill coefficient of 1.36. The predicted placebo-adjusted dose-responses of serum K+ change appeared nearly linear in the correction and maintenance phases. No clinically meaningful difference in placebo-adjusted serum K+ change from baseline at 28days was observed between maintenance regimens of SZC 5g QOD and 2.5g QD. A greater SZC treatment response was associated with high serum K+ at baseline, advanced age, lower body weight, lower estimated glomerular filtration rate, and Black/African American and Asian race, compared with the reference patient. The impact of heart failure status and diabetes status was only minor. The PopPD model of SZC adequately described changes in serum K+ concentration during correction and maintenance phase dosing. A greater treatment response was associated with various covariates, but the impact of each was modest. Overall, these findings suggest that no adjustment in SZC dose is needed for any of the covariates evaluated.

Promising aryl selenoate derivatives as antileishmanial agents and their effects on gene expression.

Leishmaniasis remains one of the main public health problems worldwide, with special incidence in the poorest populations. Selenium and its derivatives can be potent therapeutic options against protozoan parasites. In this work, 17 aryl selenoates were synthesized and screened against three species of Leishmania (Leishmania major, Leishmania amazonensis, and Leishmania infantum). Initial screening in promastigotes showed L. infantum species was more sensitive to selenoderivatives than the others. The lead Se-(2-selenocyanatoethyl) thiophene-2-carboselenoate (16) showed a half-maximal effective concentration of 3.07 µM and a selectivity index > 32.57 against L. infantum promastigotes. It was also the most effective of all 17 compounds, decreasing the infection ratio by 90% in L. infantum-infected macrophages with amastigotes at 10 µM. This aryl selenoate did not produce a hemolytic effect on human red blood cells at the studied doses (10-100 µM). Furthermore, the gene expression of infected murine macrophages related to cell death, the cell cycle, and the selenoprotein synthesis pathway in amastigotes was altered, while no changes were observed in their murine homologs, supporting the specificity of Compound 16 against the parasite. Therefore, this work reveals the possible benefits of selenoate derivatives for the treatment of leishmaniasis.

Arthrospira maxima extract prevents and cures Zika virus infection: In vitro analysis with VERO cells

Zika virus (ZIKV) infections have increased in recent years due to the lack of vaccines, specific antiviral treatments, and the difficulty controlling Aedes vectors. This infection leads to serious neurological disorders, such as microcephaly and Guillain-Barré syndrome. Although higher plants are widely recognized as sources of bioactive compounds, there is relatively limited knowledge of such molecules in algae and microalgae. In this article, we evaluated Arthrospira maxima extract for its antiviral properties against ZIKV. We performed in vitro experiments in African green monkey kidney (VERO) cells. A. maxima extract showed a strong dose-dependent inhibition of ZIKV replication. Based on the half-maximal effective concentration, microalgal extract (1.65 ± 0.08 μg/mL) was more effective than the reference drug ribavirin (3.4 ± 0.8 μg/mL). The cytotoxicity assay showed that the microalgal extract was safer than ribavirin: The selectivity index of the extract was 5 times higher than ribavirin. The microalgal extract also had a protective and preventive role in inhibiting ZIKV binding to VERO cells. After pretreating these cells with 2.5, 5, 10, or 20 μg/mL of A. maxima extract for 2 h, the highest concentration had inhibited approximately 80 % viral adsorption. Finally, to investigate the direct impacts of A. maxima extract on the virus, we incubated ZIKV with 10 μg/mL of extract before infection (−2 or − 1 h), during infection, and at 2–8 h post infection. There was unprecedented virucidal activity of A. maxima algal extract against ZIKV at all times. In conclusion, A. maxima extract counteracts viral exertion and exerts virucidal activity to prevent ZIKV infection.

Sex differences in adipose insulin resistance are linked to obesity, lipolysis and insulin receptor substrate 1.

Insulin resistance is more prominent in men than women. If this involves adipose tissue is unknown and was presently examined. AdipoIR (in vivo adipose insulin resistance index) was measured in 2344 women and 787 men. In 259 of the women and 54 of the men, insulin induced inhibition of lipolysis (acylglycerol breakdown) and stimulation of lipogenesis (glucose conversion to acylglycerols) were determined in subcutaneous adipocytes; in addition, basal (spontaneous) lipolysis was also determined in the fat cells. In 234 women and 115 men, RNAseq expression of canonical insulin signal genes were measured in subcutaneous adipose tissue. Messenger RNA transcripts of the most discriminant genes were quantified in 175 women and 109 men. Men had higher AdipoIR values than women but only when obesity (body mass index 30 kg/m2 or more) was present (p < 0.0001). The latter sex dimorphism was found among physically active and sedentary people, in those with and without cardiometabolic disease and in people using nicotine or not (p = 0.0003 or less). In obesity, adipocyte insulin sensitivity (half maximum effective hormone concentration) and maximal antilipolytic effect were tenfold and 10% lower, respectively, in men than women (p = 0.005 or less). Basal rate of lipolysis was two times higher in men than women (p > 0.0001). Sensitivity and maximum effect of insulin on lipogenesis were similar in both sexes (p = 0.26 and p = 0.18, respectively). When corrected for multiple comparison only RNAseq expression of insulin receptor substrate 1 (IRS1) was lower in men than women (p < 0.0001). The mRNA transcript for IRS1 was 60% higher in women than men (p < 0.0001). In obesity, adipose tissue insulin resistance is more pronounced in men than in women. The mechanism involves less efficient insulin-mediated inhibition of adipocyte lipolysis, increased basal rate of lipolysis and decreased adipose expression of a key element of insulin signaling, IRS1.

Novel 5-Sulfonyl-1,3,4-thiadiazole-Substituted Flavonoids as Potential Bactericides and Fungicides: Design, Synthesis, Three-Dimensional Quantitative Structure-Activity Relationship Studies.

Flavonoids, ubiquitous natural products, provide sources for drug discovery owing to their structural diversity, broad-spectrum pharmacological activity, and excellent environmental compatibility. To develop antibacterial and antifungal agents with novel mechanisms of action and innovative structures, a series of novel 5-sulfonyl-1,3,4-thiadiazole-substituted flavonoids were designed and synthesized, and their biological activities against seven agriculturally common phytopathogenic microorganisms were evaluated. The results of the antimicrobial bioassay showed that most of the target compounds displayed excellent inhibitory effects against Xanthomonas oryzae, Rhizoctonia solani, and Colletotrichum orbiculare. Compounds 1, 3, 7, 9, 13, and 14 exhibited remarkable antibacterial activity against X. oryzae pv. oryzae with EC50 values below 10 μg/mL, which were superior to bismerthiazol (70.89 μg/mL). Compound 2 (EC50 = 0.41 μg/mL) displayed the most effective inhibitory potency against R. solani in vivo, comparable protective effects with the positive control carbendizam. Preliminary mechanistic studies indicated that compound 2 induced disordered entanglement of hyphae, shrinkage of hyphal surfaces, extravasation of cellular contents, and vacuole swelling and rupture, which disrupted normal hyphal growth. Subsequently, compounds 35-53 with good antifungal activity were designed and synthesized based on reliable three-dimensional quantitative structure-activity relationship (3D-QSAR) models. Compound 49 showed high efficacy and superior antifungal activity against R. solani, with an EC50 value of 0.28 μg/mL and a half-maximal effective concentration of 0.46 μg/mL.

The fabrication method impacts biological properties of microfibrous three-dimensional in vitro cell culture scaffolds and their applications in drug screening

Two-dimensional (2D) cell culture models cannot accurately emulate the three-dimensional (3D) in vivo microenvironment; hence, the shift from compound screening in traditional 2D well plates to 3D cell culture models is forthcoming. New scaffold designs must comply with the 3Rs principle to reduce the use of animal-derived products, such as natural proteins. In line with this motivation, we engineered and compared various fibrous scaffolds featuring diverse fibre networks (coarse and fine fibres) and fibre surface properties (porous vs. smooth fibres), and tested them in in vitro cell culture. We fabricated coarse solid fibre scaffolds via a melt-based electrohydrodynamic process, which resulted in fibres with a diameter of 10.6 ± 1.9 µm and a porosity of 87.1 ± 0.9%. Using solution-based cryo-electrohydrodynamic process, we fabricated fine porous fibre and fine solid fibre scaffolds that exhibited fibres of 2.01 ± 0.5 µm and 1.72 ± 0.6 µm in diameter, with porosities of 96.5 ± 0.3% and 94.4 ± 0.5%, respectively. The scaffolds underwent only chemical modification via an alkaline treatment to enhance their hydrophilicity. We used mouse fibroblast L929 cells and human triple-negative breast cancer MDA-MB-231 cells as cell models, and used the MTT assay to measure their viability. After seven days of cell culture, all scaffolds were thoroughly penetrated. However, the coarse scaffolds created by the melt-based electrohydrodynamic process and the porous fibre scaffolds produced by the solution-based cryo-electrohydrodynamic process outperformed the simpler morphology of smooth fine fibres. Furthermore, the 50% half-maximum effective concentration (EC50) of doxorubicin tested in human glioblastoma (U87-MG) and human triple-negative breast cancer (MDA-MB-231) cell lines in 3D cell culture models was approximately twice as high as that achieved in conventional 2D cell cultures. Therefore, we infer that poly (ɛ-caprolactone) (PCL) microfibre scaffolds, devoid of animal-derived product surface treatment, may provide a favourable 3D milieu for various cell types for in vitro studies.

Ultrasonic-assisted extraction and UHPLC determination of ascorbic acid, polyphenols, and half-maximum effective concentration in Citrus medica and Ziziphus spina-christi fruits using multivariate experimental design

This study aimed to determine the concentrations of ascorbic acid and polyphenols in fruits and peels of Citrus medica and Ziziphus spina-christi grown in Ethiopia. Conditions of ultrasound-assisted extraction (UAE) and ultra-high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD) were optimized, using a multivariate experimental design. The optimum conditions of UAE were 15 min extraction time at 35 ℃, with 75 % aqueous methanol as solvent, and a fruit powder-to-solvent ratio (m/v) of 1:15. Among the different drying conditions investigated, freeze-drying was found to be appropriate for analyzing ascorbic acid, polyphenols, and antioxidant potential. The overall ranges, across the fruits and peels, of ascorbic acid, total polyphenols, and antioxidant potentials (EC50) obtained were 8.7 ± 1.4–91.2 ± 2.6 mg/100 g, 253.0 ± 6.3–764.1 ± 25.8 mg GAE/100 g and 2.4 ± 0.1–26.1 ± 2.9 mg/mL, respectively. This indicates that the fruits and peels of the studied plants are advantageous as sources of ascorbic acid and polyphenols.

A mini-review of the anti-SARS-CoV-2 potency of Amaryllidaceae alkaloids

BackgroundNature has perennially served as an infinite reservoir of diverse chemicals with numerous applications benefiting humankind. In recent years, due to the emerging COVID-19 pandemic, there has been a surge in studies on repurposing natural products as anti-SARS-CoV-2 agents, including plant-derived substances. Among all types of natural products, alkaloids remain one of the most important groups with various known medicinal values. The current investigation focuses on Amaryllidaceae alkaloids (AAs) since AAs have drawn significant scientific attention as anti-SARS-CoV-2 agents over the past few years. Purpose and study designThis study serves as a mini-review, summarizing recent advances in studying the anti-SARS-CoV-2 potency of AAs, covering two aspects: structure-activity relationship and mechanism of action (MOA). MethodsThe study covers the period from 2019 to 2023. The information in this review were retrieved from common databases including Web of Science, ScienceDirect, PubMed and Google scholar. Reported anti-SARS-CoV-2 potency, cytotoxicity and possible biological targets of AAs were summarized and classified into different skeletal subclasses. Then, the structure-activity relationship (SAR) was explored, pinpointing the key pharmacophore-related structural moieties. To study the mechanism of action of anti-SARS-CoV-2 AAs, possible biological targets were discussed. ResultsIn total, fourteen research articles about anti-SARS-CoV-2 was selected. From the SAR point of view, four skeletal subclasses of AAs (lycorine-, galanthamine-, crinine- and homolycorine-types) appear to be promising for further investigation as anti-SARS-CoV-2 agents despite experimental inconsistencies in determining in vitro half maximal inhibitory effective concentration (EC50). Narciclasine, haemanthamine- and montanine-type skeletons were cytotoxic and devoid of anti-SARS-CoV-2 activity. The lycorine-type scaffold was the most structurally diverse in this study and preliminary structure-activity relationships revealed the crucial role of ring C and substituents on rings A, C and D in its anti-SARS-CoV-2 activity. It also appears that two enantiomeric skeletons (haemanthamine- and crinine-types) displayed opposite activity/toxicity profiles regarding anti-SARS-CoV-2 activity. Pharmacophore-related moieties of the haemanthamine/crinine-type skeletons were the substituents on rings B, C and the dioxymethylene moiety. All galanthamine-type alkaloids in this study were devoid of cytotoxicity and it appears that varying substituents on rings C and D could enhance the anti-SARS-CoV-2 potency. Regarding MOAs, initial experimental results suggested Mpro and RdRp as possible viral targets. Dual functionality between anti-inflammatory activity on host cells and anti-SARS-CoV-2 activity on the SARS-CoV-2 virus of isoquinoline alkaloids, including AAs, were suggested as the possible MOAs to alleviate severe complications in COVID-19 patients. This dual functionality was proposed to be related to the p38 MAPK signaling pathway. ConclusionOverall, Amaryllidaceae alkaloids appear to be promising for further investigation as anti-SARS-CoV-2 agents. The skeletal subclasses holding the premise for further investigation are lycorine-, crinine-, galanthamine- and homolycorine-types.

The antioxidant and anticancer activity of Quercus coccifera plant leaves extracts

Quercus species are one of the medicinal plants that commonly used in the treatment of different diseases. Quercus coccifera (Q. coccifera) is part of the Quercus species which grow in Jordan and used in traditional folklore medicine. The aim of this study is to confirm the ability of (Q. coccifera) leaves extracts to exert anticancer activity.In this study, an extraction method of the dried-leaves using different polarity solvents was used. Extracts were pre-evaluated for antioxidant and anticancer activities while active extracts were used to measure half maximal effective concentration (EC50) against 2,2-Diphenyl-1-picrylhydrazyl (DPPH), and Half-maximal inhibitory concentration (IC50) against cancer cells.Methanol, boiled and microwaved water extracts had greater than 80 % antioxidant activity, and the strongest activity, of more than 99 %, was boiled water extract. Similarly, the pre-evaluation treatments of cancer cell lines indicated a strong biological activity of more than 70 % from the previously mentioned extracts, and the highest activity, of greater than 90 %, was from boiled water extracts against all cancer cell lines. The highest EC50 against DPPH was obtained by using 0.009 mg/ml boiled water extracts, which was lower than positive control quercetin. In the same manner, lung, breast, and prostate cancer cell lines were highly affected by boiled water extracts with IC50 of 14.1, 7.2, and 25.1 µg/ml, respectively, and a selectivity index (SI) of greater than 4.71.Q. coccifera leaves extracts show promising ability to be a source of a new anticancer therapeutics.

Synthesis and Biological Evaluation of Telmisartan Alkylamine Derivatives as Potential Anticancer Agents.

Telmisartan is an angiotensin II receptor type 1 (AT1) antagonist with anticancer properties against solid and hematological cancer cell lines. Using telmisartan as a template, we developed alkylamine derivatives with reduced AT1 activity but increased anticancer activity. Synthesis of candidate compounds was carried out via hexafluorophosphate benzotriazole tetramethyl uronium coupling reaction, then their inhibition of cell proliferation was determined via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and colony-formation assay was carried out on the lead candidate compound 8 Cell death via apoptosis or necrosis by compound 8 was determined by flow cytometry using annexin V and propidium iodide, tolerability dosing was carried out in ICR mice, and tumor-reduction properties were determined in an MDA-MB-231 xenograft model. Some of the synthesized candidates exhibited good inhibition of cell proliferation with low micromolar half maximal effective concentrations in triple-negative breast cancer cell lines MDA-MB-231 and 4T1. Compound 8 exhibited lower affinity towards AT1 than parent telmisartan, inhibition of colony formation, and cell-cycle analysis revealed apoptosis as potentially important in causing cell death. In vivo evaluation with compound 8 indicated that it was well tolerated at high concentrations in healthy mice. Additionally, compound 8 showed higher growth inhibition in the MDA-MB-231 tumor xenograft mouse model compared to telmisartan. Our study indicated that alkylamine derivatives of telmisartan exhibited good solubility and higher inhibition of cancer cell proliferation than telmisartan. Compound 8 was found to be a good lead compound, with potential for development as an anticancer agent.

Small-molecule positive allosteric modulation of homomeric kainate receptors GluK1-3: development of screening assays and insight into GluK3 structure.

The kainate receptors GluK1-3 (glutamate receptor ionotropic, kainate receptors 1-3) belong to the family of ionotropic glutamate receptors and are essential for fast excitatory neurotransmission in the brain, and are associated with neurological and psychiatric diseases. How these receptors can be modulated by small-molecule agents is not well understood, especially for GluK3. We show that the positive allosteric modulator BPAM344 can be used to establish robust calcium-sensitive fluorescence-based assays to test agonists, antagonists, and positive allosteric modulators of GluK1-3. The half-maximal effective concentration (EC50) of BPAM344 for potentiating the response of 100 μm kainate was determined to be 26.3 μm for GluK1, 75.4 μm for GluK2, and 639 μm for GluK3. Domoate was found to be a potent agonist for GluK1 and GluK2, with an EC50 of 0.77 and 1.33 μm, respectively, upon co-application of 150 μm BPAM344. At GluK3, domoate acts as a very weak agonist or antagonist with a half-maximal inhibitory concentration (IC50) of 14.5 μm, in presence of 500 μm BPAM344 and 100 μm kainate for competition binding. Using H523A-mutated GluK3, we determined the first dimeric structure of the ligand-binding domain by X-ray crystallography, allowing location of BPAM344, as well as zinc-, sodium-, and chloride-ion binding sites at the dimer interface. Molecular dynamics simulations support the stability of the ion sites as well as the involvement of Asp761, Asp790, and Glu797 in the binding of zinc ions. Using electron microscopy, we show that, in presence of glutamate and BPAM344, full-length GluK3 adopts a dimer-of-dimers arrangement.

Novel quinoline derivatives with broad-spectrum antiprotozoal activities.

Several quinoline derivatives incorporating arylnitro and aminochalcone moieties were synthesized and evaluated in vitro against a broad panel of trypanosomatid protozoan parasites responsible for sleeping sickness (Trypanosoma brucei rhodesiense), nagana (Trypanosoma brucei brucei), Chagas disease (Trypanosoma cruzi), and leishmaniasis (Leishmania infantum). Several of the compounds demonstrated significant antiprotozoal activity. Specifically, compounds 2c, 2d, and 4i displayed submicromolar activity against T. b. rhodesiense with half-maximal effective concentration (EC50 ) values of 0.68, 0.8, and 0.19 µM, respectively, and with a high selectivity relative to human lung fibroblasts and mouse primary macrophages (∼100-fold). Compounds 2d and 4i also showed considerable activity against T. b. brucei with EC50 values of 1.4 and 0.4 µM, respectively.

Chemo-рroteomic analysis of the pharmacological properties of vitamin В12 derivatives

Background. Chemical derivatives of vitamin B12 are characterized by a wide range of pharmacological effects. It is important to learn how to establish relationships between changes of the corrin ring structure in vitamin B12 derivatives and changes in pharmacological properties.Objective: to evaluate the interaction of six vitamin B12 derivatives (aquacobalamin, diaquacobinamide, aquacyano-forms of heptaethanolamine, heptaethylenediamine, heptamethyl and heptabutyl cobyrinates) with human proteome proteins.Material and methods. Using the method of chemoinformational (chemoproteomic) analysis, implemented within the framework of algebraic recognition theory and topological data analysis, the constants of half maximal inhibitory concentration (IC50) and half maximal effective concentration (EC50) of human proteome proteins were assessed.Results. Significant differences were found in the interactions of the studied molecules with 1200 proteins. It was shown that the chemoproteomic profiles of each of the compounds form three groups of molecules with similar proteomic properties: (1) aquacobalamin, (2) diaquacobinamide, aquacyano-forms of heptaethanolamine and heptaethylenediamine cobyrinates, (3) aquacyano-forms of heptamethyl and heptabutyl cobyrinates. A more detailed analysis of the chemoproteomic profiles of the studied compounds using the GO (Gene Ontology) nomenclature of biological functions of proteins made it possible to identify functional GO categories indicating differences in the biological effects of the studied compounds: neuroprotective regulation of neurotransmitter activity (serotonin receptor activity, cholinergic synapses, regulation of dopamine secretion, receptor thyroid hormones), reduction of inflammation (inhibition of cytokine biosynthesis, including tumor necrosis factor alpha and interleukin 1 beta, I-kappa-B kinases / nuclear factor kappa В, leukocyte migration), etc.Conclusion. Based on the obtained data conclusions were drawn about the potential effects and safety of the studied substances.

Structure-Guided Discovery of PD-1/PD-L1 Interaction Inhibitors: Peptide Design, Screening, and Optimization via Computation-Aided Phage Display Engineering.

Cancer immunotherapy harnesses the immune system to combat tumors and has emerged as a major cancer treatment modality. The PD-1/PD-L1 immune checkpoint modulates interactions between tumor cells and T cells and has been extensively targeted in cancer immunotherapy. However, the monoclonal antibodies known to target this immune checkpoint have considerable side effects, and novel PD-1/PD-L1 inhibitors are therefore required. Herein, a peptide inhibitor to disrupt PD-1/PD-L1 interactions was designed through structure-driven phage display engineering coupled to computational modification and optimization. BetaPb, a novel peptide library constructed by using the known structure of PD-1/PD-L, was used to develop inhibitors against the immune checkpoint, and specific peptides with high affinity toward PD-1 were screened through enzyme-linked immunosorbent assays, homogeneous time-resolved fluorescence, and biolayer interferometry. A potential inhibitor, B8, was preliminarily screened through biopanning. The binding affinity of B8 toward PD-1 was confirmed through computation-aided optimization. Assessment of B8 variants (B8.1, B8.2, B8.3, B8.4, and B8.5) demonstrated their attenuation of PD-1/PD-L1 interactions. B8.4 exhibited the strongest attenuation efficiency at a half-maximal effective concentration of 0.1 μM and the strongest binding affinity to PD-1 (equilibrium dissociation constant = 0.1 μM). B8.4 outperformed the known PD-1/PD-L1 interaction inhibitor PL120131 in disrupting PD-1/PD-L1 interactions, revealing that B8.4 has remarkable potential for modification to yield an antitumor agent. This study provides valuable information for the future development of peptide-based drugs, therapeutics, and immunotherapies for cancer.