Pharmacological Evaluation Research Articles

UHPLC‐QTOF‐MS‐Based Metabolic Profiling, Estimation of Antioxidants and Anticancer Potential Assessment of Dicliptera bupleuroides Nees by Caspase Activation, and ROS and MMP Evaluation by Using Fluorescence Microscopy

ABSTRACTDicliptera bupleuroides has been used traditionally for treating various ailments in many countries. Its detailed phytochemical profiling and pharmacological evaluation in treating cancer based on reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) analysis along with caspase activation studies has not been done previously. This study was conducted for detailed phytochemical profiling of D. bupleuroides by using ultra‐high performance liquid chromatography‐quadrupole time‐of‐flight mass spectrometry (UHPLC‐QTOF‐MS) and to evaluate its antioxidant and anticancer potential against hepatocellular carcinoma (HepG2) cells. The extract was subjected to UHPLC‐QTOF‐MS analysis for metabolite profiling, and GNPS Molnet enhancer determined its major phytochemical classes. Phytochemical tests and multimode antioxidant assays were also executed. The extract was pharmacologically evaluated for its anticancer potential against HepG2 cells using CCK‐8 assay. Intracellular ROS generation and loss in MMP were studied by fluorescence microscopy. Network pharmacology and molecular docking of selected compounds were executed. The extract contains total phenolic content as 58.37 ± 0.32 μg GAE/mg extract, and remarkable antioxidant potential was revealed in total antioxidant contents (TAC) and total reducing power (TRP) assays, i.e., 70.4 ± 0.4, 83.2 ± 1.9 μg ascorbic acid equilant (AAE) Borges/mg extract. The UHPLC‐QTOF‐MS analysis showed that the extract contains flavonoid glycosides, phenolic acids, flavans, O‐methylated flavonoids, linoleic acids, terpene glycosides, triterpenoid saponins, and certain other phytoconstituents. Maximum inhibition of 77.13% ± 1.60% against HepG2 cells was observed at 24 h. Loss in MMP by JC‐1 (5,5′,6,6′‐tetrachloro‐1,1′,3,3′‐tetraethylbenzimi‐ dazolylcarbocyanine iodide) staining and increased ROS generation, and caspase‐3, caspase‐8 activation by extract significantly illustrated the cancer cell death by apoptosis. D. bupleuroides could be a potential source of safe and therapeutically active compounds that can be purified to develop safer, selective, and efficacious anticancer agents with fewer side effects.

Pharmacological Evaluation of Respiratory Safety Following a Single Intravenous Administration of Theophylline in Sprague-Dawley Rats.

Drinking tea is an important cultural and sensory activity in modern society. However, tea consumption may influence biological functions because of its high polyphenol and methylxanthine content. Theophylline is a methylxanthine that plays a significant role in respiratory physiology and is used clinically as a bronchodilator. In this study, we assessed the pharmacological safety of theophylline by investigating its effects on the respiratory responses of experimental rodents. Six-week-old male Sprague-Dawley rats were intravenously administered 0, 2.5, 5, or 10 mg/kg theophylline. After systemic theophylline exposure, the respiratory rate, tidal volume, and minute volume were monitored and statistically analyzed. There was a significant increase in respiratory rate and minute volume with no change in tidal volume at theophylline doses of >5 mg/kg. Changes in respiratory rate and minute volume peaked at 0.25 h after exposure; the respiratory rate increased by 58.2% and 88.9%, and the minute volume increased by 50.5% and 72.8% with the 5 and 10 mg/kg doses, respectively, compared with the vehicle control. These results provide fundamental scientific data that can be used in clinical applications.

Isolation and Bioassay of Linear Veraguamides from a Marine Cyanobacterium (Okeania sp.)

Marine cyanobacteria have gained momentum in recent years as a source of novel bioactive small molecules. This paper describes the structure elucidation and pharmacological evaluation of two new (veraguamide O (1) and veraguamide P (2)) and one known (veraguamide C (3)) analogs isolated from a cyanobacterial collection made in the Las Perlas Archipelago of Panama. We hypothesized that these compounds would be cytotoxic in cancer cell lines. The compounds were screened against HEK-293, estrogen receptor positive (MCF-7), and triple-negative breast cancer (MDA-MB-231) cells as well as against a broad panel of membrane-bound receptors. The planar structures were determined based on NMR and MS data along with a comparison to previously isolated veraguamide analogs. Phylogenetic analysis of the collection suggests it to be an Okeania sp., a similar species to the cyanobacterium reported to produce other veraguamides. Veraguamide O shows no cytotoxicity (greater than 100 μM) against ER-positive cells (MCF-7) with 13 μM IC50 against MDA-MB-231 TNBC cells. Interestingly, these compounds show affinity for the sigma2/TMEM-97 receptor, making them potential leads for the development of non-toxic sigma 2 targeting ligands.

Quality Consistency Evaluation of Chemical Composition and Pharmacology of Pheretima aspergillum Dispensing Granules and Traditional Decoction Based on HPLC, GC-IMS, and Animal Experiments.

This study investigates the consistency of quality between the traditional decoction (TD) of Pheretima aspergillum and its dispensing granule decoction (DGD) by examining their chemical composition and antithrombotic efficacy. We first determined the nucleoside components, protein content, and volatile compounds in 10 batches of TD and 9 batches of DGD sourced from three manufacturers using high-performance liquid chromatography (HPLC), UV-vis spectrophotometry, and gas chromatography ion mobility spectrometry (GC-IMS). Next, we assessed antithrombotic efficacy by measuring the relative length of tail thrombosis and calculating the thrombus inhibition rates at 24 and 48h. Our comprehensive analysis revealed that the concentrations of hypoxanthine, uridine, inosine, adenosine, and proteins in TD were significantly higher than those in DGD. Moreover, the types and concentrations of volatile compounds in TD and DGD exhibited substantial differences, with 19 volatile compounds identified as differentially present between the two decoctions. Despite these compositional differences, both decoction types demonstrated equivalent antithrombotic efficacy. Overall, the quality of DGD from manufacturers A and B aligned closely with that of TD. However, TD exhibited a higher overall quality compared to DGD, while both decoctions maintained consistent antithrombotic efficacy.

Abstract WP364: Translational Relevance and Versatility of the Thromboembolic Stroke Model in Mice: Implications for Pharmacological and Safety Evaluations

Abstract WP364: Translational Relevance and Versatility of the Thromboembolic Stroke Model in Mice: Implications for Pharmacological and Safety Evaluations

Design, synthesis, and pharmacological evaluation of triazine-based PI3K/mTOR inhibitors for the potential treatment of non-small cell lung cancer.

Dysregulated activation of the PI3K/AKT/mTOR pathway is crucial in the development of cancer, and disrupting it could potentially lead to cancer suppression, making it a viable strategy for cancer treatment. Here, as a consecutive work of our team, we described the identification and optimization of PI3K/mTOR inhibitors based on triazine scaffold, which exhibited potent PI3K/mTOR inhibitor activity. The systematically structure-activity relationship (SAR) results demonstrated that compound 5nh displayed high efficacy against PI3Kα and mTOR, with the IC50 values of 0.45nM and 2.9nM, respectively. Importantly, compared to the lead compound PKI-587, 5nh demonstrated significant inhibitory activity against non-small-cell lung cancer (NSCLC) cell lines, particularly HCC-827, with a 43-fold increase (3.5nM vs 150nM). Additionally, the compound showed effective inhibition against the EGFR-resistant variant HCC-827(GR) cell line. Mechanism validation demonstrated that 5nh significantly interfered with the PI3K/AKT/mTOR signaling pathway in HCC-827cells. Furthermore, the oral pharmacokinetic properties of 5nh had been observably improved, with AUC0-t and Cmax increasing by 13-16 times at a dose of 10mg/kg in mice. Importantly, the in vivo efficacy study demonstrated that orally treatment of 5nh led to significant tumor growth suppression, with a TGI value of 84.4%. Collectively, our systematically medicinal chemistry campaigns suggested that 5nh, a novel oral available triazine derivative, held promise as a candidate for therapy of NSCLC by targeting the PI3K/AKT/mTOR cascade.

Assessment of the Properties and Mechanism of Three Lotus (Nelumbo nucifera Gaertn.) Parts for Sleep Improvement.

Lotus leaves (LL), petals (LP), and seeds (LS) are believed to have properties that can improve sleep. However, their efficacy in improving sleep has not been fully validated. This study aimed to investigate the multitarget mechanisms of extracts from these lotus parts for sleep improvement using chemical analysis, bioactivity assessment, meta-analysis, network pharmacology evaluation, and molecular docking. The chemical components in LL, LP, and LS were detected using ultra-high-performance liquid chromatography-ultraviolet-quadrupole time-of-flight tandem mass spectrometry, their total flavonoid and phenolic contents were determined, and their 2,2-diphenyl-1-picrylhydrazyl scavenging activity was estimated. Subsequently, relevant literature was collected for meta-analysis. Finally, potential targets were predicted using network pharmacology, and the results were validated by molecular docking. 48 compounds were identified from the lotus extracts. LL had the highest total flavonoid content, while LS had the highest total phenolic content. Extracts from both parts exerted significant antioxidant effects. A meta-analysis identified the potential of lotus to improve sleep. Armepavine, asimilobine, nuciferin, and luteolin were identified as key components, and AKT1, EGFR, DRD2, and PIK3R1 were revealed as key targets. PI3K-Akt was identified as a key signaling pathway. This study provides an important reference for studying the role of the lotus in improving sleep quality.

Dimeric dipeptide mimetics of BDNF loop 4 are potential antidepressants with novel mechanisms of action

This review covers original research focused on the design, synthesis, and pharmacological evaluation of an innovative dimeric dipeptide mimetic of brain-derived neurotrophic factor (BDNF) loop 4 bis-(N-monosuccinyl-L-seryl-L-lysine) hexamethylenediamide (GSB-106). Developed using a proprietary approach for creating low-molecular-weight neurotrophin mimetics, GSB-106 displayed marked antidepressant-like activity following systemic and oral administration. The article details its pharmacological properties in vitro and in vivo, mechanism of action, development of a tablet form of the dipeptide, and findings from toxicological and pharmacokinetic studies.

Exploring the Therapeutic Potential of Benzimidazole Hybrids as Antimicrobial Agents: An In-Depth Analysis

Background: Heterocyclic compounds play an essential role in biological sys-tems and occur widely in nature. They are fundamental in the development of pharma-ceuticals aimed at combating microbial infections and other biological activities. Phar-macological evaluations have demonstrated their efficacy against diverse bacterial strains. This study investigates the antimicrobial properties of various benzimidazole hy-brids. The findings highlight the significant influence of substituting nitrogenous scaf-folds with various heteroatoms on the potential development of new antimicrobial agents. Objective: This review article is expected to make a substantial contribution to the ad-vancement of efficacious antibacterial medications. The research's goal is to improve the efficacy of combating bacterial infections by utilizing the potent properties of benzimid-azole-based hybrid scaffolds. In the end, this will aid in reducing the global incidence of this contagious illness. Methods: Several nitrogen-containing heterocyclic compounds display substantial poten-tial as antibacterial agents. These compounds possess fused benzene and imidazole nu-clei. These nuclei could change the number of electrons they have, which in turn affects their physiochemical characteristics. The versatility of drugs arises from their capacity to interact with receptors in various modalities, which is a key factor in drug pharmacolog-ical screening. Benzimidazole-based hybrids have demonstrated a wide range of pharma-cological effects, including antibacterial, anti-HIV, anticancer, antimalarial, antiviral, an-tifungal, antioxidant, anti-inflammatory, and anti-tubercular activities. Results: Pyrazole, imidazole, oxazole, thiazole, indole, and benzimidazole are examples of compounds that include nitrogen species. These nitrogen-containing compounds en-gage in metabolic interactions with other molecules within the cell. Nevertheless, an over-abundance of reactive nitrogen species can cause cytotoxicity, causing harm to vital bio-logical macromolecules. But benzimidazole is traditionally the most effective, with a wide range of important qualities, including antibacterial, anti-HIV, anticancer, antima-larial, antiviral, antifungal, antioxidant, anti-inflammatory, and anti-tubercular activities. Conclusion: This study focuses on the efficacy of novel benzimidazole-based hybrid scaffolds in inhibiting microbial growth or eradicating microorganisms. The study pri-marily focuses on recent studies carried out from 2009 to 2024. The study highlights the effectiveness of different benzimidazole-based hybrids using MIC values. More in-depth studies also show that adding electron-withdrawing groups (EWGs) to the nitrogenous framework might make them more effective. Further research is necessary to design strong, least-toxic benzimidazole-based hybrids that can either kill or inhibit multidrug-resistant (MDR) bacteria.

Isolation and Bioassay of Linear Veraguamides from a Marine Cyanobacterium ( Okeania sp. ).

Marine cyanobacteria have gained momentum in recent years as a source of novel bioactive small molecules. This paper describes the structure elucidation and pharmacological evaluation of two new, veraguamide O ( 1 ) and veraguamide P ( 2 ), and one known, veraguamide C( 3 ), analogs isolated from a cyanobacterial collection made in the Las Perlas Archipelago of Panama. We hypothesized that these compounds would be cytotoxic in cancer cell lines. The compounds were screened against HEK-293, estrogen receptor positive (MCF-7), and triple-negative breast cancer (MDA-MB-231) cells as well as against a broad panel of membrane bound receptors. The planar structures were determined based on NMR and MS data along with comparison to previously isolated veraguamide analogs. Phylogenetic analysis of the collection suggests it to be an Okeania sp., a similar species to the cyanobacterium reported to produce other veraguamides. Veraguamide O shows no cytotoxicity (greater than 100 M) against ER positive cells (MCF-7) with 13 M IC 50 against MDA-MB-231 TNBC cells. Interestingly, these compounds show affinity for the sigma2/TMEM-97 receptor making them potential leads for development of non-toxic sigma 2 targeting ligands.

Pharmacological Evaluation of Anti-depressant activity of Aegle Marmelos ethanolic leaves extract in mice

Bael (A. marmelos) is an important medicinal herb which has been and extensively used in Ayurveda, Siddha and other medicinal systems. The aim of this study is to evaluate the pharmacological activity of leaves part of the plant Aegle marmelos. Ethanolic extract of Aegle marmelos leaves prepared by successive extraction technique. This research carroed out to find out the Percentage Yield of Extract of Aegle marmelos, determination of LD50 of the EEAML as per OECD guidelines and to investigate the anti-depressant effect of Aegle marmelos leaves extract using tail suspension test (TST) and forced swim test (FST) parameters at dose levels of 200 mg/kg and 400 mg/kg respectively in albino swiss mice. Aegle marmelos leaves extract showed significant antidepressant activity probably due to GABA facilitatory action of phytoconstituents such as flavonoids, tannic acid, marmesinin, phenols, saponin etc. Results suggested that cortisol and corticosterone responded differently to severe stressors with cortisol being a quicker responder than corticosterone. Keywords: Aegle Marmelos, phytoconstituents, Anti-depressant activity

Design, Synthesis, and Pharmacological Evaluation of Nonsteroidal Tricyclic Ligands as Modulators of GABAA Receptors.

GABAA receptors (GABAARs) are the major elements of inhibitory neurotransmission in the central nervous system (CNS). They are established targets for regulation by endogenous brain neuroactive steroids (NASs) such as pregnanolone. However, the complexity of de novo synthesis of NAS derivatives has hindered attempts to circumvent the principal limitations of using endogenous NASs, including selectivity and limited oral bioavailability. In this study, we designed a series of tricyclic compounds, inspired by the structures of pregnanolone and pregnenolone sulfate, to explore novel nonsteroidal alternatives. Using patch clamp electrophysiology, we demonstrate that these compounds exhibit either positive or negative allosteric modulation of GABAARs. Specifically, we discover a positive allosteric modulator (PAM) and a series of tricyclic sulfate-based negative allosteric modulators (NAMs) all active at the micromolar level. This research has significantly broadened the chemical diversity of ligands targeting GABAARs offering potential for efficacious allosteric modulators while avoiding the complexity of NAS synthesis.

Spinal Involvement of TRPV1 and PI3K/AKT/mTOR Pathway During Chronic Postoperative Pain in Mice.

Chronic postoperative pain (CPOP) is among the main consequences of surgical procedures, directly affecting the quality of life. Although many strategies have been used to treat this symptom, they are often ineffective. Thus, studies investigating CPOP-associated mechanisms may help to develop more effective treatment strategies. Therefore, the present study investigated the spinal participation of the transient potential receptor vanilloid type 1 (TRPV1) and PI3K/AKT/mTOR pathway activation during CPOP. In this study C57BL/6 male mice were used, and CPOP was induced by muscle retraction and incision. The nociceptive threshold was measured by the von Frey filament test. For pharmacological evaluation, TRPV1 and PI3K/AKT/mTOR inhibitors were administered intrathecally. TRPV1 and PI3K/AKT/mTOR protein levels were evaluated by Western blotting. The results showed that CPOP increased TRPV1 and mTOR protein levels, and pretreatment with the specific inhibitors alleviated CPOP. In addition, pretreatment with the TRPV1 antagonist SB-366791 attenuated mTOR protein levels. The results suggest that TRPV1 and the PI3K/AKT/mTOR pathway are involved in CPOP at the spinal level, and TRPV1 may activate mTOR during this process.

Uncovering the Mechanisms of BaBaoWuDanYaoMo Against Influenza A Virus and Virus-Induced Inflammation Based on Network Pharmacology and Pharmacological Evaluation.

The pro-inflammatory response triggered by influenza viruses can contribute to viral pneumonia, even death. The effect and mechanism of BaBaoWuDanYaoMo (BB) against influenza virus remains obscure. To predict its therapeutic targets via network pharmacology and verify the therapeutic effect and the mechanism of BB against influenza virus in vitro and in vivo. The potential active and underlying mechanism of BB in the treatment of influenza virus was predicted through network pharmacological strategies and Molecular Docking. The protective and anti-inflammatory effects of BB were determined by cytopathic effect (CPE), quantitative real-time PCR, mitochondrial membrane potentials and Western blotting assay in vitro. BALB/c mice were intranasally infected with A/PR/8/34 (H1N1) (PR8) and orally administrated BB (500 mg/kg, 250 mg/kg and 125 mg/kg) or oseltamivir daily. The normal group was orally administrated PBS for 5 days. Lung indexes, histological changes and cytokines were determined on the 6th day. BB could effectively inhibit A/Puerto Rico/8/1934 (H1N1), A/GZ/GIRD07/09 (H1N1), A/HK/Y280/97 (H9N2) and A/Aichi/68 (H3N2) with IC50 values of 116.5 ± 2.2, 59.86 ± 8.33, 102.87 ± 6.66 and 66.43 ± 6.785μg/mL, respectively. It could inhibit PR8-induced apoptosis and inhibit the expression of apoptosis markers (cleaved PARP). BB inhibited the mRNA expression of MCP-1/CCL-2, IL-6, CXCL-8/IL-8, TNF-α and CXCL-10/IP-10, and downregulated the protein expression of phosphorylated AKT/p38 and TLR3 in vitro. BB (500 and 250 mg/kg) could improve pulmonary histopathological changes, decrease the lung index and suppress the mRNA expression of CXCL1/KC, TNF-α, CXCL9/MIG and IL-1β in vivo. BB has a protective effect on PR8-induced acute lung injury (ALI) probably via inhibition of apoptosis process and interfering with TLR3, p38 MAPK and PI3K-AKT signaling pathways. This study provided a potential treatment for influenza from BB, and network pharmacology provided a strategy to explore active components and mechanism of TCMs against influenza virus infection.

Chemical diversity of the herbal decoction of Plectranthus ornatus and its anti-nociceptive and anti-inflammatory activities in zebrafish models.

Plectranthus ornatus is a medicinal plant originally from Africa but adapted to Brazil's climate conditions. It is recognized for its therapeutic properties, particularly for treating liver and stomach diseases, gastritis control, heartburn, and hangover. Therefore, studies on its chemical composition and pharmacological evaluation are important for the safe use of the plant. To investigate the chemical composition and pharmacological activity of leaf decoction of P. ornatus. The lyophilized herbal decoction from P. ornatus leaves was extracted with MeOH (methodology A), and CHCl3 and n-BuOH (methodology B). The compounds were isolated using chromatographic techniques. Their structures were determined using spectroscopic methods (1H and 13C NMR, IR, and HRESIMS) and comparison with published data. The lyophilized herbal decoctions DPO A (decoction from methodology A) and DPO B (methodology B), and compounds 8, 9, 15, and 16 (4, 20, and 40 mg/kg) were evaluated for their toxicity, anti-nociceptive, and anti-inflammatory effects in experimental adult zebrafish (Danio rerio) models. The present study focused on the CHCl3 and MeOH soluble fractions from the lyophilized leaf decoction leading to the isolation of five new diterpenes (1, 4-7) and two new glucuronide flavonoid derivatives (2 and 3). In addition, four diterpenes (14-17), two glucuronide flavonoid derivatives (10 and 11), three phenolics (8, 12, and 13), and one disaccharide (9) previously reported were also isolated. In zebrafish essays, all samples showed no toxicity and exhibited an anti-nociceptive effect in at least one of the tested doses: 9, 15, 16, and DPO B (4 mg/kg), 8 (20 mg/kg), and DPO A (40 mg/kg). Moreover, the compounds 15 (4, 20, and 40 mg/kg) and 16 (4, 20, and 40 mg/kg) exhibited anti-inflammatory effects. The lyophilized decoctions (DPO A and DPO B) including the compounds 8, 9, 15, and 16 exhibited significant anti-nociceptive effects in adult zebrafish and showed no toxicity. Since pain can be a symptom of liver, stomach, and gastrointestinal disorders, and all the samples proved to be non-toxic, the herbal decoction of P. ornatus leaf could be considered a potential therapeutic option in pain management, supporting the ethnopharmacological use of the plant.

Synthesis, pharmacological evaluation, and molecular docking studies of some 1,3,4-oxadiazolyl-5-yl thiones bearing halo-nitrophenyl hydrazides derivatives as antioxidant and antimicrobial agents

Synthesis, pharmacological evaluation, and molecular docking studies of some 1,3,4-oxadiazolyl-5-yl thiones bearing halo-nitrophenyl hydrazides derivatives as antioxidant and antimicrobial agents

Anticoagulants: From chance discovery to structure-based design.

Taking a historical perspective, we review the discovery, pharmacology, and clinical evaluation of the old and new anticoagulants that have been approved for clinical use. The drugs are discussed chronologically, starting in the 1880s, and progressing through to 2024. The innovations in technology used to develop novel anticoagulants came in fits and starts and reflected the advances in science and technology over these decades, whereas the shift from anecdote to evidence-based use of anticoagulants was delayed until the principles of epidemiology and biostatistics were introduced into clinical trial design and to the approval process. Hirudin, heparin, and vitamin K antagonists were discovered by chance, and were used clinically before their mechanism of action was elucidated and before their net clinical benefits were evaluated in randomized clinical trials. Subsequent anticoagulants were designed based on a better understanding of the structure and function of coagulation proteins, including antithrombin, thrombin, and factor Xa, and underwent more rigorous preclinical and clinical evaluation before regulatory approval. By simplifying oral anticoagulation, the direct oral anticoagulants have revolutionized anticoagulation care and have enhanced the uptake of anticoagulation, but bleeding has not been eliminated and there is a need for more effective and convenient anticoagulants for thrombosis triggered by the contact pathway of coagulation. The newly developed factor XIa and XIIa inhibitors have the potential to address these unmet clinical needs and are undergoing clinical evaluation for several indications. SIGNIFICANCE STATEMENT: Anticoagulant therapy is the cornerstone of treatment and prevention of thrombosis, which remains a leading cause of morbidity and mortality worldwide. Elucidation of the structure and function of coagulation enzymes, their cofactors, and inhibitors, coupled with advances in structure-based design led to the discovery of more convenient, safer, and more effective anticoagulants that have revolutionized the management of thrombotic disorders.

Systems pharmacology and experimental evaluation to investigate the therapeutic targets of Chinese medicine QiShenYiQi in the treatment of ischemic stroke

Systems pharmacology and experimental evaluation to investigate the therapeutic targets of Chinese medicine QiShenYiQi in the treatment of ischemic stroke

Design, synthesis and pharmacological evaluation of triazolopyrimidines derivatives as novel agonists for benzodiazepine receptor

Design, synthesis and pharmacological evaluation of triazolopyrimidines derivatives as novel agonists for benzodiazepine receptor

Synthesis, Characterization, and in vitro pharmacological evaluation of Zinc(II) complexes of cycloalkanes and bioactive nitrogen donor heterocycles

Synthesis, Characterization, and in vitro pharmacological evaluation of Zinc(II) complexes of cycloalkanes and bioactive nitrogen donor heterocycles