Development Of New Drugs Research Articles

Therapeutic potential of rutin in premenstrual depression: evidence from in vivo and in vitro studies

IntroductionPremenstrual dysphoric disorder (PMDD) is a cyclical mood disorder that severely affects the daily life of women of reproductive age. Most of the medications being used clinically have limitations such as low efficacy, side effects, and high cost, so there is an urgent need to discover safer and more effective medications. Rutin is a natural flavonol glycoside with various pharmacological properties including antidepressant. The study of the efficacy and mechanism of action of rutin in PMDD-depressed subtype model rats plays an important role in the discovery of new drugs for the treatment of PMDD.MethodsBinding of rutin to gamma-aminobutyric acid type A receptors (GABAA receptors) was probed using molecular docking, microscale thermophoresis, radioactive receptor ligand binding assay and cell membrane clamp experiment. Behavioral tests in mice were performed to screen the optimal dose of rutin. Behavioral tests were performed to evaluate the effects of rutin on depressed mood, memory impairment, and social impairment in PMDD-depressed subtype model rats. HE staining and Golgi staining were performed to observe the neuronal damage in rat hippocampus. UHPLC-MS/MS targeted metabolomics was performed to detect the changes of neurotransmitter content in rat hippocampus. PCR array to detect the effect of rutin on mRNA expression of GABAA receptor partial subunits in rat hippocampus.ResultsThe docking score of rutin with the GABAA receptor benzodiazepine site was −11.442 and the gliding score was −11.470. The Kd of rutin with the GABAA receptor (α1β2γ2) was 1.17 ± 0.89 μM. Rutin competed with [H3]-flunitrazepam for the GABAA receptor benzodiazepine site and inhibited the inward flow of chloride ions (P < 0.05). In PMDD-depressed subtype rats, rutin alleviated depressed mood, memory impairment and social impairment, ameliorated hippocampal neuronal damage and reduces gamma-aminobutyric acid (GABA) and acetylcholine (ACh) levels (P < 0.05). Moreover, we found that rutin did not affect the relative mRNA expression of GABAA receptor subunits in rat hippocampus.DiscussionOverall, rutin alleviated depressed mood, memory impairment and social impairment in PMDD-depressed subtype rats, which may be related to binding to GABAA receptor benzodiazepine sites, inhibiting chloride ions inward flow, ameliorating hippocampal neuronal damage and reducing GABA and ACh levels. The results of this study provide an experimental basis and scientific evidence for the development of new drugs for the treatment of PMDD.

Global longitudinal strain as an early marker of cardiac damage after cardiotoxic medications, a state of the art review.

Ejection fraction (EF) is the principal parameter used clinically to assess cardiac function and provides prognostic information. However, significant myocardial damage can be present despite preserved EF. Recently, the measurement of left ventricle (LV) deformation by global longitudinal strain (GLS) has been introduced as a novel early marker of cardiac dysfunction. Cardiotoxicity is a frequent side effect of several drugs most notably those used in the treatment of cancer. Although oncology drugs remain the best known cardiotoxic medications, many other drugs can potentially affect LV function. The early recognition of LV dysfunction due to cardiotoxicity is important and of increasing clinical relevance particularly with the rapid pace of development of new drugs. The aim of our review is to provide an overview of the current literature regarding utility of GLS to assess drug-induced myocardial damage. We propose that GLS is a sensitive early marker of myocardial dysfunction associated with the use of certain medications with high risk of cardiotoxicity. Thus, the use of this technique can potentially alert the clinician to myocardial toxicity before reductions in EF are seen.

Plackett-Burman Design Combined with Response Surface Methodology to Recover Polysaccharides with Cardiovascular Protective Potential from Waste Zizania latifolia Bracts.

Zizania latifolia is the second aquatic vegetable in China. The circular valorization of its waste bracts remains ongoing concern. In this work, the cellulase-microwave-assisted extraction (CMAE) of polysaccharides from waste Z. latifolia bracts (PWZLBs) was explored. Seven parameters were selected via single-factor test, of which three significant parameters were screened out using Plackett-Burman design, followed by response surface methodology optimization. The optimal CMAE for PWZLBs were: cellulase addition of 0.5%, microwave time of 7 min, and microwave power of 425 W, resulting in a yield of 0.82 ± 0.08%. Four polysaccharide fractions (PWZLBs-1 ~ 4) were isolated from PWZLBs, of which PWZLBs-1 accounted for major proportion and exerted higher scavenging capacities on diphenyl picryl hydrazinyl and hydroxyl radicals. More importantly, PWZLBs-1 elicited anticoagulation via prolonging thrombin time and prothrombin time, exhibiting potential for cardiovascular protection. Various characterizations confirmed that PWZLBs-1 is a heteropolysaccharide containing uronic acids and sulfates, with galactose (34.3%) as the predominant monosaccharide, and has a molecular weight of 8061 kDa. This work provides clues for the circular valorization of waste Z. latifolia bracts and offers potential opportunities for the development of new cardiovascular protective drugs.

Prospects for the development of new drugs for the treatment of osteoporosis

With the population aging, the prevalence of osteoporosis is rapidly increasing, which has become serious public health problem. At present, there is a severe shortage of effective anti-osteoporotic drugs in China, making it difficult to control the growing trend of osteoporosis. The key pathological mechanism of osteoporosis is bone homeostasis imbalance. It is necessary to continuously study the bone cell biology, important pathway regulating bone homeostasis, and its regulatory network constituted by bone-muscle-vessel-immune system-nerve-gut microbiota, in order to identify key targets for new drug, and develop more novel effective drugs to increase bone mineral density, improve bone strength, reduce fracture risk and protect bone health.

Atractylodin alleviates polycystic ovary syndrome by inhibiting granule cells ferroptosis through pyruvate dehydrogenase kinase 4-mediated JAK-STAT3 pathway.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder, and its close relationship with oxidative stress has been well-documented. Atractylodin (ATR) plays a role in the treatment of many diseases through its antioxidant function. However, its function in PCOS remains unexplored. In this study, the function and underlying mechanisms of ATR in mitigating PCOS symptoms were investigated. A mouse model of PCOS induced using DHEA and a high-fat diet was established, and many factors such as hormone levels (FSH, LH, testosterone, and progesterone), the estrous cycle, and ovarian shape were evaluated. In vitro, PCOS model was established by DHEA-induced KGN cell, and the effects of ATR on ferroptosis and oxidative stress markers were explored. Specifically, the viability of KGN cells treated with ATR was assessed using the CCK-8 assay, and the levels of malondialdehyde (MDA), glutathione (GSH), and reactive oxygen species (ROS) were measured to evaluate oxidative stress. Expression of ferroptosis-related genes (NRF2, GPX4, SLC7A11) and PDK4 was analyzed by qRT-PCR and Western blotting. PDK4's interaction with ATR was examined through molecular docking and confirmed by surface plasmon resonance (SPR) analysis. Our data show that the treatment of ATR markedly increased hormone levels and improved normal estrous cycles. Moreover, ATR was found to improve ovarian morphology by decreasing cystic dilatation and increasing the number of corpora lutea. Mechanistically, our research found that ATR regulates the expression of PDK4 by binding to its GLY331 and inhibits granulosa cell ferroptosis by regulating the JAK-STAT3 pathway mediated by PDK4. In conclusion, our study suggest that ATR may be a therapeutic option for managing PCOS and PDK4 could be a target for the development of new drugs for PCOS.

Progress in the Study of TAp73 and Sperm Apoptosis.

The study of the mechanism of oligoasthenospermia, which is a major cause of male infertility, has been the focus of research in the field of male reproduction. TAp73, a member of the p53 family of oncogenes, is endowed with tumor-suppressing activity due to its structural and functional homology with p53. It has been found that TAp73, plays a key role in spermatogenesis and maintaining male reproduction. When TAp73 is low-expressed or absent, the process of spermatogenesis is severely impaired, and mice deficient in TAp73 exhibit spermatogonial DNA damage, disturbed apical cytoplasmic specialization, and spermatocyte malformations resulting in reduced male fertility. Nevertheless, when TAp73 is overexpressed, it not only drives exogenous death receptors to regulate germ cell apoptosis, but also interacts with its various substrate proteins to promote the translocation of cytoplasmic Bax proteins to the mitochondria, resulting in the upregulation of the Bax/Bcl-2 ratio on the mitochondrial membrane and triggering a series of mitochondrial apoptotic effects. In this article, we will analyze the mechanism of TAp73 and sperm apoptosis, and elaborate the mechanism of TAp73 upregulation, exogenous apoptosis pathway and mitochondrial apoptosis pathway to systematically explain that the process of apoptosis induced by high expression of TAp73 is not fixed and single, but is interconnected, so as to provide a basis for the treatment of oligoasthenospermia and the research and development of new drugs using TAp73 as a target.

A Review on Pyrazole Derivatives Used in the Treatment of Rheumatoid Arthritis: Recent Advancement and Drug Development.

Rheumatoid arthritis (RA) is an autoimmune disorder where inflammation and destruc-tion of bone are the hallmarks of the disease. This review focuses on the etiology, pathophysiolo-gy, and treatment strategies for RA, along with the different approaches used for the synthesis of pyrazoles, the characterization of various properties, and their biological significance for curing RA. The activated immune system of the body causes inflammation of the synovial joint due to the interaction of immune cells, such as T and B lymphocytes, macrophages, plasma cells, den-dritic cells and mast cells. The treatment for RA has been revolutionized with the discovery of new chemical compounds and an understanding of their mechanism in the treatment of the dis-ease. Pyrazoles are the starting materials for the synthesis of heterocyclic compounds and possess great relevance in the pharmaceutical field for the development of new drugs. They are versatile bio-scaffolds in medicinal chemistry and organic synthesis. This has been followed by a deep analysis of pyrazoles and their derivatives on the basis of medical significance in the treatment of RA. This follow-up and information may help the chemists, scientists, and researchers to generate new pyrazole compounds with high efficacy for better treatment of patients with RA. We summa-rize the review with an understanding of the core of pyrazoles and a claim that their derivatives may be helpful in the development of efficient drugs against RA.

Advances in antibacterial agents for Mycobacterium fortuitum.

Mycobacterium fortuitum is an emerging human pathogen, characterized by an increase in prevalence and antibacterial resistance over the years, highlighting the need for the development of new drugs against this rapidly growing nontuberculous mycobacterium (NTM). To support this crusade, this review summarizes findings from the past two decades concerning compounds with antimycobacterial activity against M. fortuitum. It identifies the most promising and effective chemical frameworks to inspire the development of new therapeutic alternatives for infections caused by this microorganism. Most compounds effective against M. fortuitum are synthetic, with macozinone, featuring a 2-piperazine-benzothiazinone framework, standing out as a notable drug candidate. Among natural products, the polyphenolic polyketide clostrubin and the sansanmycin peptide analogs have shown efficacy against this NTM. Some compounds' mechanisms of action on M. fortuitum have been studied, including NITD-916, which acts as an enoyl-acyl carrier protein reductase inhibitor, and TBAJ-5307, which inhibits F-ATP synthase. Moreover, this review discusses the pathogenic molecular mechanisms and potential therapeutic targets within this mycobacterium.

Isolation and biomimetic synthesis of acylphloroglucinol meroterpenoids as anti-breast cancer agents from Dryopteris crassirhizoma.

Two new acylphloroglucinol-nerolidol meroterpenoids (APNMs) [(±)-1 and (±)-2], with a skeleton of mixed sesquiterpene and dimeric acylphloroglucinol biosynthetic origin, were isolated from the medicinal pteridophyte Dryopteris crassirhizoma. Inspired by the proposed biosynthetic pathway, we initially completed the biomimetic syntheses of eight optically active Dryopteris APNMs (1-8) in a one-pot domino reaction. The structures of APNMs 1-8 including their absolute configurations were unambiguously established by a combination of NMR analysis, ECD calculations, and synthetic methods. Furthermore, an underlying method based on NMR data to assign the stereochemistry of the long-chain alcohol moieties in APNMs was revealed. Anti-triple negative breast cancer (anti-TNBC), antifungal and antibacterial activities of the synthetic meroterpenoids were evaluated. All tested compounds exhibited cytotoxicity against TNBC cells MDA-MB-231 with IC50 values in the range of 1.22-27.43 μM, with (-)-8 being the most potent antitumor agent (IC50: 0.48 μM; selectivity index: 32.04). This study resulted in the biomimetic synthesis of APNMs for the first time and positioned APNMs from D. crassirhizoma as a new class of promising candidates for the development of new anti-TNBC drugs.

Insights into in vitro and in silico studies of α-glucosidase inhibitors isolated from the leaves of Grewia optiva (Malvaceae).

α-Glucosidase plays a critical role in glucose metabolism by breaking down complex carbohydrates into simpler sugars for intestinal absorption. Due to the side effects of current α-glucosidase inhibitors, there is increasing interest in exploring alternative therapeutic options. Inspired by the traditional uses of Grewia optiva J.R.Drumm. ex Burret (Malvaceae family) as an anti-diabetic herb, we isolated gnaphaffine A (1), a cyclic glycosylated homolignan, together with kaempferol derivatives (trans-tiliroside 2, cis-tiliroside 3, and astragalin 4) from the ethyl acetate fraction. In vitro antioxidant assays revealed that 1 exhibited anti-DPPH• and anti-ABTS+• activity (IC50 of 39.42 and 52.84μg/mL, respectively), comparable to ascorbic acid (IC50 of 43.34 and 47.56μg/mL, respectively). Moreover, 1 demonstrated a seven-fold stronger inhibition of α-glucosidase activity than acarbose (IC50 of 8.2 and 57.8μg/mL, respectively). Importantly, 1 was non-toxic to AC16 normal cardiomyocyte cell lines. Computational analyses identified two key factors contributing to the α-glucosidase inhibitory activity of 1: (a) hydrogen bonding interactions with catalytic residues (E277 and D352) and (b) a calculated ∆Gbind of -51.20kcal/mol. Furthermore, 3 showed the most favorable in silico binding profile, with the highest ∆Gbind (-55.89kcal/mol) and higher hydrogen bond occupancy compared to 1 and 2. These findings suggest that 1 and 3 may serve as promising lead compounds for the development of new α-glucosidase drugs.

The effect and mechanism of sanguinarine against PCV2 based on the analysis of network pharmacology and TMT quantitative proteomics.

Porcine circovirus type 2 (PCV2) is highly prevalent in nature and serves as the primary pathogen responsible for porcine circovirus-associated disease (PCVD/PCVAD), posing a significant threat to pig production. Currently, vaccination alone could not provide the complete protection for PCV2 infection. The active ingredients of traditional Chinese medicine have shown a positive effect in combating viral infections. This study employed tandem mass tag (TMT) labeled proteomic analyses and network pharmacology to examine the effect and mechanism of sanguinarine against PCV2. IFIH1, IFITM1, p38α, and JNK were identified as the key targets of sanguinarine against PCV2 based on proteomics and network pharmacology. Using PCV2-infected PK-15 cells, it was discovered that sanguinarine inhibited the expression of the PCV2 CAP gene by upregulating IFIH1, thereby promoting STAT1 phosphorylation and activating MAVS expression. This, in turn, facilitated IRF3 phosphorylation, leading to increase IFITM1 expression. Simultaneously, sanguinarine suppressed the expression of the PCV2 CAP gene by inhibiting the expression of p38α, JNK, and p-JNK protein. In conclusion, the results of this study suggest that sanguinarine exerts anti-PCV2 effects through different targets and pathways, which lays the foundation for the subsequent development of new anti-PCV2 veterinary drugs.

New C-linked diarylheptanoid dimers as potential α-glucosidase inhibitors evidenced by biological, spectral and theoretical approaches.

Diabetes mellitus (DM) is a chronic metabolic disorder characterized by elevated blood glucose levels, generally due to defects of insulin action or secretion. Inhibition of α-glucosidase, an enzyme responsible for carbohydrate degradation, is a promising strategy for managing postprandial hyperglycemia in diabetic patients. In this study, two new C-linked diarylheptanoid dimers, kaemgalanganols A (1) and B (2), were isolated from K. galanga, which showed obvious inhibitory activity on α-glucosidase but weak activity on protein tyrosine phosphatase 1B (PTP1B). Kaemgalanganol B had an IC50 value of 35.1μM against α-glucosidase, obviously more potent than kaemgalanganol A (IC50=78.5μM) and acarbose (IC50=363.0μM). Enzyme kinetic study indicated that 2 was a reversible mixed-type inhibitor of α-glucosidase via non-competitive and anti-competitive inhibition modes. Fluorescence quenching and UV-visible spectroscopic study revealed that fluorescence quenching mechanism of 2 on α-glucosidase is a combination of dynamic quenching and static quenching, accompanied by non-radiative energy transfer. Compound 2 formed complex with α-glucosidase closer to the Tyr residue, and induced changes in both the microenvironment and peptide backbone. Surface hydrophobicity and CD spectra measurement indicated that 2 affected the function of α-glucosidase by decreasing the surface hydrophobicity of α-glucosidase as well as altering the secondary structure instead of the overall three-dimensional framework, which is consistent with the results of fluorescence experiment. Molecular docking manifested that compound 2 had a strong binding affinity (-7.27kcal/mol) with α-glucosidase, higher than 1 (-9.82kcal/mol) and acarbose (-4.48kcal/mol), consistent with the enzyme inhibitory assay. Besides hydrogen bonds, electrostatic interactions and hydrophobic interactions played important roles in the binding of 2 with α-glucosidase. This study disclosed the inhibitory activity and mechanism of 2 against α-glucosidase, which provides a theoretical basis for the development of new antidiabetic drugs form K. galanga.

Design, synthesis, and evaluation of dehydroabietyl imidazolidine-2,4-diones, 2,4,5-triones, and 2-thioxoimidazolidine-4,5-diones as TDP1 inhibitors and dual TDP1/TDP2 inhibitors.

Tyrosyl DNA phosphodiesterases 1 and 2 (TDP1 and TDP2), which are enzymes involved in the repair of DNA, are regarded as promising targets for the development of new anticancer drugs. In this study, a series of imidazolidine-2,4-diones, 2,4,5-triones, and 2-thioxoimidazolidine-4,5-diones based on dehydroabietylamine (DHAAm) were synthesized. The inhibitory activity of the new compounds against TDP1 and TDP2, as well as their cytotoxic characteristics, were evaluated. All types of heterocyclic DHAAm derivatives demonstrated effective inhibition of TDP1 in the micromolar range, with IC50 values in the range of 0.63-4.95 µM. It was observed that only the 2-thioxoimidazolidine-4,5-diones were TDP2 inhibitors, representing the first class of dual TDP1/TDP2 inhibitors among DHAAm derivatives. The findings of this study may contribute to an enhanced comprehension of the subsequent design of novel dual TDP1/TDP2 inhibitors for the further development of new antitumor agents.

Unlocking the potential of Traditional Chinese Medicine (TCM): Shipi Xiaoji formula (SPXJF) as a novel ferroptosis inducer in hepatocellular carcinoma.

Hepatocellular Carcinoma (HCC) is a major health concern with limited treatment options. Traditional Chinese Medicine (TCM) offers potential therapeutic approaches for HCC, and SPXJF, a TCM formula, has shown promise in clinical observations for prolonging the survival of liver cancer patients. To investigate the anti-tumor effects of SPXJF on HCC cells and explore its potential mechanism, focusing on ferroptosis induction. LC/Q-TOF-MS was used for compound identification. Cell viability assays, EdU proliferation assay, colony formation assay, wound healing assay, Transwell assay, and Western-blotting were conducted to evaluate the effects of SPXJF on HCC cell proliferation, migration, and invasion. Bioinformatics analysis and RT-PCR were employed to identify potential ferroptosis-related genes and validate the results. Ferroptosis induction was investigated using ferroptosis inhibitors, ROS and lipid peroxidation detection, and TEM. In vivo experiments using a subcutaneous xenograft tumor model confirmed the anti-tumor effects of SPXJF and its ability to induce ferroptosis in HCC. SPXJF effectively inhibited the proliferation, migration, and invasion of HCC cells in vitro. The mechanism of action was found to be related to the induction of ferroptosis, as evidenced by increased intracellular Fe2+ and ROS levels, decreased GSH levels, altered mitochondrial morphology, and upregulation of ferroptosis-inducing proteins ACSL4 and LPCAT3, along with downregulation of ferroptosis-inhibiting proteins xCT and GPX4. Bioinformatics analysis and RT-PCR further identified GSTZ1, CDC25A, AURKA, NOX4, and CAPG as potential ferroptosis-related genes regulated by SPXJF. In vivo experiments confirmed the anti-tumor effects of SPXJF and its ability to induce ferroptosis in HCC. SPXJF exerts anti-tumor effects on HCC cells by inducing ferroptosis, and its mechanism of action involves the regulation of ferroptosis-related genes and proteins. This study provides a theoretical basis for the clinical treatment of HCC and the development of new anti-cancer drugs, offering a valuable contribution to the field of ethnopharmacology.

Triple-helix β-glucan-based self-assemblies, synthesis, characterization and anticarcinogenic effect.

Triple negative breast cancer (TNBC) seriously endangers women's life and health due to its high invasion and mortality. Reactive oxygen species (ROS) mediated tumor cells apoptosis is considered an effective anticancer approach. Herein, we designed a natural active triple helix β-Glucan (BFP) wrapped single walled carbon nanotubes (SWNTs)-loaded doxorubicin (DOX) self-assembly (BSD) via generating excess ROS to induce oxidative stress damage for TNBC therapy. BSD could directly consume glutathione (GSH) to promote ROS. In vitro results demonstrated that BSD exhibited obvious antitumor effects to breast cancer cells by promoting apoptosis. Un-targeted metabolomics under molecular level identified the specific metabolic targets and unveiled that BSD markedly disturbed multiple metabolic pathways, including purine metabolism, pentose phosphate pathway, glutathione metabolism pathways, amino sugar and nucleotide sugar metabolism and energy metabolism, led to the inhibition of DNA and RNA synthesis, the generation of ROS, the exacerbation of DNA damage, the disruption of cell membrane integrity and the decrease of ATP. In vitro and in vivo oxidative stress assays further verified that BSD significantly promoted intracellular oxidative stress and resulted in cell damage. This study provides theoretical basis for the development and screening of new drugs based on ROS therapy for TNBC.

Establishment and optimization of a novel mouse model of hyperuricemic nephropathy

Hyperuricemia is a metabolic disorder characterized by elevated serum uric acid levels. Soluble urate can activate immune responses, and the excessive accumulation of urate in the kidneys results in hyperuricemic nephropathy (HN). However, the lack of an established HN model is a major obstacle to advancing research into the pathogenesis of HN and the development of novel drugs. In this study, we generated and evaluated an optimized mouse model of HN by the combined administration of potassium oxonate and hypoxanthine at various dosages. Our results demonstrated that intraperitoneal injection of 200 mg/kg potassium oxonate with gavage of 500 mg/kg hypoxanthine caused renal injury in mice, as evidenced by the elevation in serum uric acid, serum creatinine, and 24 h albuminuria levels, as well as pathological changes in renal histology. Intraperitoneal injection of 200 mg/kg potassium oxonate with gavage of 500 mg/kg hypoxanthine markedly increased the production of uric acid, inhibited uricase activity, and disrupted uric acid transporters. This led to supersaturated urate deposition in the kidneys, triggering renal inflammation and fibrosis, thereby promoting HN progression. In conclusion, we successfully established a stable and efficient mouse model that can mimic the pathogenesis of HN. This novel model may facilitate the discovery of therapeutic targets and the development of new drugs for the treatment of HN.

Expose the bioactive properties of Picrorhiza kurroa root extract oil (PKEO): phytochemical composition and therapeutic activities

Background: The present study aims to explore the bioactive properties of essential Oil (PKEO) derived from Picrorhiza kurroa (commonly known as kutki), a medicinal plant known for its therapeutic potential. Picrorhiza kurroa essential oil has a distinct chemical profile, which sets it apart from other essential oils. The bioactive compounds present in Picrorhiza kurroa essential oil may lead to the development of new drugs, particularly for treating inflammatory and oxidative stress-related disorders. The research aims to study the extraction, phytochemical composition, and various biological activities of PKEO. Methodology: Oil obtained through hydro-distillation contains various phytochemical compounds, including steroids, triterpenoids, alkaloids, phenols, proteins, flavonoids, and tannins. Its bioactivity and aroma are attributed to its phenolic and sesquiterpene esters. Results and Discussion: The total phenolic content is 250.47 μg GAE/g, and the total flavonoid content is 245.26 μg QE/g. UV-visible and IR spectroscopic analyses confirm the presence of phenolic and terpenoid ester functional groups. PKEO has moderate antioxidant activity, with IC50 values of 98.19 µg/mL in DPPH scavenging and 42.72% inhibition in the ABTS assay. It also exhibits dose-dependent inhibition of protein denaturation and HRBC stabilizing activity. Antimicrobial tests show PKEO inhibits E. coli growth, indicating potential antibacterial properties. Conclusion: These findings highlight PKEO's promising bioactive profile, suggesting potential therapeutic and cosmetic formulation applications. The antifungal activity also shows the potential antifungal effects of the PKEO.

Fast and Efficient Screening and Separation Based on AUF‐MS Combined With Molecular Docking Technology and Network Pharmacology Method for Potential Xanthine Oxidase Inhibitors From Pinelliae Rhizoma Praeparatum

ABSTRACTIntroductionStudies show that Pinelliae Rhizoma Praeparatum (PRP) has some pharmacological effects in enhancing immunity and against gout.ObjectivesA mathematical model was created for extraction process optimization, analysis and identification, activity screening, and isolation and purification; moreover, the mechanism of action was studied.MethodsFirst, the extraction of PRP was investigated using the gray wolf optimization mathematical regression model; the extraction variables were optimized to maximize the yield. Second, we used network pharmacological analysis to predict potential targets for PRP in treating gout; xanthine oxidase inhibitors (XODIs) were rapidly screened using AUF‐MS and enzyme‐catalyzed reaction kinetics. The potential antigout effects of the obtained active substances were verified using molecular docking and molecular dynamics simulation analysis. Finally, with activity screening as the guide, an HSCCC method combined with consecutive injection using the UNIFAC mathematical model and semipreparative HSCCC was successfully developed for the separation and purification of XODIs.ResultsThe results verified that uridine, guanosine, adenosine, liquiritin, and liquiritigenin of PRP exhibited high biological affinity toward XOD.ConclusionThis study clarifies the mechanisms of action of a medicinal plant of interest at the molecular level and can provide more opportunities for the discovery and development of new therapeutic drugs from other food resources.

Uncovering the Mechanism of Action of Antiprotozoal Agents: A Survey on Photoaffinity Labeling Strategy

Plasmodium, Leishmania, and Trypanosoma parasites are responsible for infectious diseases threatening millions of people worldwide. Despite more recent efforts devoted to the search for new antiprotozoal agents, efficacy, safety, and resistance issues still hinder the development of suited therapeutic options. The lack of robustly validated targets and the complexity of parasite’s diseases have made phenotypic screening a preferential drug discovery strategy for the identification of new chemical entities. However, via this approach, no information on biological target(s) and mechanisms of action of compounds are provided. Among the target deconvolution strategies useful to fill this gap, photoaffinity labeling (PAL) has emerged as one of most suited to enable investigation in a complex cellular environment. More recently, PAL has been exploited to unravel the molecular basis of bioactive compounds’ function in live parasites, allowing elucidation of the mechanism of action of both approved drugs and new chemical entities. Besides highlighting new potential drug targets, PAL can provide valuable information on efficacy and liabilities of small molecules at the molecular level, which could be exploited to greatly facilitate the rational optimization of compounds in terms of potency and safety. In this review, we will report the most recent studies that have leveraged PAL to disclose the biological targets and mechanism of action of phenotypically active compounds targeting kinetoplastid diseases (i.e., human African trypanosomiasis, leishmaniasis, and Chagas disease) and malaria. Moreover, we will comment on potential perspectives that this innovative approach can provide in aiding the discovery and development of new antiprotozoal drugs.

Quantitative Live Imaging Reveals Phase Dependency of PDAC Patient-Derived Organoids on ERK and AMPK Activity.

Patient-derived organoids represent a novel platform to recapitulate the cancer cells in the patient tissue. While cancer heterogeneity has been extensively studied by a number of omics approaches, little is known about the spatiotemporal kinase activity dynamics. Here we applied a live imaging approach to organoids derived from 10 pancreatic ductal adenocarcinoma (PDAC) patients to comprehensively understand their heterogeneous growth potential and drug responses. By automated wide-area image acquisitions and analyses, the PDAC cells were non-selectively observed to evaluate their heterogeneous growth patterns. We monitored single-cell ERK and AMPK activities to relate cellular dynamics to molecular dynamics. Furthermore, we evaluated two anti-cancer drugs, a MEK inhibitor, PD0325901, and an autophagy inhibitor, hydroxychloroquine (HCQ), by our analysis platform. Our analyses revealed a phase-dependent regulation of PDAC organoid growth, where ERK activity is necessary for the early phase and AMPK activity is necessary for the late stage of organoid growth. Consistently, we found PD0325901 and HCQ target distinct organoid populations, revealing their combination is widely effective to the heterogeneous cancer cell population in a range of PDAC patient-derived organoid lines. Together, our live imaging quantitatively characterized the growth and drug sensitivity of human PDAC organoids at multiple levels: in single cells, single organoids, and individual patients. This study will pave the way for understanding the cancer heterogeneity and promote the development of new drugs that eradicate intractable cancer.