Biochemical Targets Research Articles

Metformin as a potential antidepressant: Mechanisms and therapeutic insights in depression.

Depression is one of the most disabling psychiatric disorders, whose pathophysiology has not been fully understood. Increasing numbers of preclinical studies have highlighted that metformin, as the first-line hypoglycaemic agent, has a potential pleiotropic effect on depression. Moreover, there is emerging evidence that metformin shows antidepressant activity and improves depressive symptoms in rodent models of depression. However, the exact role and underlying mechanism of metformin in depression remain unclear and still need to be investigated. Recent studies suggest that metformin not only improves neuronal damage and structural plasticity in the hippocampus but also enhances the antidepressant effect of antidepressants. Therefore, in this review, we summarize the existing evidence for the use of metformin as a psychopharmaceutical and elaborate on the underlying mechanisms of metformin in mitigating the onset and progression of depression, as well as the associated biochemical signaling pathways and targets involved in the pathogenesis of depression. After reviewing several studies, we conclude that metformin helps reduce depressive symptoms by targeting multiple pathways, including the regulation of neurotransmitters, enhanced neurogenesis, anti-inflammatory effects, and changes in gut microbiota. We aim to gain a deeper understanding of the mechanism of action of metformin and provide new insights into its clinical value in the prevention and therapy of depression.

Biochemical targets of chick embryos affected by sub-lethal concentrations of lambda-cyhalothrin and imidacloprid.

The insecticides Lambda-cyhalothrin (LCT) and imidacloprid (IMD) are extensively utilized in Egyptian agriculture. Embryonic chicken is a readily accessible model organism commonly employed in various studies. Eggs of (Gallus Gallus) chicken were immersed in an aqueous solution of two sub-lethal concentrations (0.375 and 0.0375mg/L for LCT; 0.05 and 0.005mg/L for IMD) for 30 sec on the fourth day of incubation of chick embryos. Significant reductions of acetylcholinesterase (AChE) activity of brain 18- and 21-day chicks were observed in the groups treated with LCT and IMD dependent on concentrations. There were significant changes (reduction or enhancement) in serum activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) by LCT and IMD at the used concentrations, while non-significant stimulation in the AST/ALT ratio (AAR) was found. In 18th - day embryos, the activities of glutathione S-transferase (GST) and acid phosphatase (ACP) were not significantly changed by LCT but were significantly increased by IMD. Liver alkaline phosphatase (ALP) activity showed no significant change except IMD at 0.05mg/L. However, serum enzyme activity was significantly reduced in all groups. In addition, the tested insecticides caused notable increases in the creatinine and total protein content. The protein profile; proteins separation with sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed an increase in four bands that are consistent with the results of the biomarkers level. Findings indicate that even the pesticide's low concentrations are not safe and may lead to severe damage to the embryos and may lead to significant harm or developmental disruption in the embryos.

Neuromodulating Alkaloids from Millipede Defensive Secretions.

Millipedes have long been known to produce structurally diverse chemical defenses, including hydrogen cyanide, terpenoid alkaloids, and oxidized aromatics. Although the hydrogen cyanide and oxidized aromatic producing millipedes have been well studied, less than 10% of the terpenoid alkaloid producers have been chemically investigated. Several previous studies have shown that alkaloids disorient predators, but their biochemical target is currently unknown. Herein, we investigated the defensive secretions of a colobognath millipede, Ischnocybe plicata, and elucidated the constitution, absolute configuration, and conformation of four new highly oxidized terpenoid alkaloids, termed ischnocybines, using a range of analytical techniques. The ischnocybines are actively secreted from the defensive glands and were shown to disorient ants, a likely common predator. Evaluation of the ischnocybines in a panel of neuroreceptors revealed that ischnocybine A possesses potent (Ki 13.6 nM) and selective (100-fold) binding affinity for sigma-1, an orphan neuroreceptor, over sigma-2. These molecules represent the most complex alkaloids to be discovered from millipedes and provide the first potential insights into a biochemical target responsible for their defensive properties.

Preclinical delayed toxicity studies of BCMA CAR T-cell injection in B-NDG mice with multiple myeloma.

Based on the efficacy data from the previous study of B-cell maturation antigen (BCMA) chimeric antigen receptor (CAR) T-cell injection, we further examined the delayed toxicity for 8 weeks after a single dose of BCMA CAR T-cell injection to observe possible toxic reactions. B-NDG mice transplanted with multiple myeloma (MM) cells were given a single dose of BCMA CAR T-cell injection at two dosages or human normal T cells and then subjected to examinations including clinical signs, weight and food intake measurements, haematology, blood biochemical analysis, cytokine assay, T-lymphocyte subpopulation quantification and histopathology on days 28 and 56 after dosing. In addition, quantitative polymerase chain reaction (qPCR) was used to quantify DNA fragments in different tissues to assess the tissue distribution of CAR and provide a basis for its preclinical safety evaluation and clinical dosing. In the delayed toxicity study, no mortality or significant toxic effects such as reductions in food intake, body weight, relevant biochemical parameters and target organ weights were observed in the BCMA CAR T-cell-treated groups. Compared to the model group, restorative changes in clinical signs and clinicopathology indicating therapeutic effects were seen in the BCMA CAR T-cell-treated groups. Human-derived cytokines interleukin-2 (IL-2), IL-4, IL-6, IL-12, IL-10, tumor necrosis factor α (TNF-α), and interferon-γ (IFN-γ) could be detected in all cancer cell-bearing mice by cytokine level measurement. IFN-γ levels showed a geometric increase due to the graft versus host disease (GVHD) response induced in the mice, while the levels of the other cytokines did not show significant changes. Histopathological examination indicated that the BCMA CAR T-cell treatment groups showed mixed cellular infiltration of human-derived T cells, cancer cells, and inflammatory cells in several target organs including the liver, spleen, lung, and kidney, and some of them showed mild tissue damage, but the number of the animals and the severity of damage were significantly less than those of the T-cell control group as well as the model group. The results of the tissue distribution study showed that BCMA CAR T cells were mainly concentrated in the kidney, lung, bone marrow and the related immune organs/tissues, and the distribution of BCMA CAR T cells was highly consistent with that of MM cells, suggesting that BCMA CAR T cells could follow the cancer cells during metastatic targeting of the tissues. The present study demonstrated a low toxicity of BCMA CAR T-cell injection, with manageable side effects and good anticancer activity and without observable adverse effects. This study provides data to support future clinical studies of BCMA CAR T-cell injection for MM.

Cannabinoid-Inspired Inhibitors of the SARS-CoV-2 Coronavirus 2'-O-Methyltransferase (2'-O-MTase) Non-Structural Protein (Nsp10-16).

The design and synthesis of antiviral compounds were guided by computationally predicted data against highly conserved non-structural proteins (Nsps) of the SARS-CoV-2 coronavirus. Chromenephenylmethanone-1 (CPM-1), a novel biphenylpyran (BPP), was selected from a unique natural product library based on in silico docking scores to coronavirus Nsps with high specificity to the methyltransferase protein (2'-O-MTase, Nsp10-16), which is responsible for viral mRNA maturation and host innate immune response evasion. To target the 2'-O-MTase, CPM-1, along with intermediate BPP regioisomers, tetrahydrophenylmethanones (TPMs), were synthesized and structurally validated via nuclear magnetic resonance (NMR) data and DP4+ structure probability analyses. To investigate the activity of these BPPs, the following in vitro assays were conducted: SARS-CoV-2 inhibition, biochemical target validation, mutagenicity, and cytotoxicity. CPM-1 possessed notable activity against SARS-CoV-2 with 98.9% inhibition at 10 µM and an EC50 of 7.65 µM, as well as inhibition of SARS-CoV-2's 2'-O-MTase (expressed and purified) with an IC50 of 1.5 ± 0.2 µM. In addition, CPM-1 revealed no cytotoxicity (CC50 of >100 µM) or mutagenicity (no frameshift or base-pair mutations). This study demonstrates the potential of computational modeling for the discovery of natural product prototypes followed by the design and synthesis of drug leads to inhibit the SARS-CoV-2 2'-O-MTase.

Exploring the 3,5-Dibromo-4,6-dimethoxychalcones and Their Flavone Derivatives as Dual α-Glucosidase and α-Amylase Inhibitors with Antioxidant and Anticancer Potential.

The rising levels of type 2 diabetes mellitus (T2DM) and the poor medical effects of the commercially available antidiabetic drugs necessitate the development of potent analogs to treat this multifactorial metabolic disorder. It has been demonstrated that targeting two or more biochemical targets associated with the onset and progression of diabetes along with oxidative stress and/or cancer could be a significant strategy for treating complications related to this metabolic disorder. The 3,5-dibromo-4,6-dimethoxychalcones (2a-f) and the corresponding flavone derivatives (3a-f) were synthesized and characterized using spectroscopic (NMR, HR-MS and FT-IR) techniques. The inhibitory effect of both series of compounds against α-glucosidase and α-amylase was evaluated in vitro through enzymatic assays. Selected compounds were also evaluated for potential to activate or inhibit superoxide dismutase. Compound 3c was selected as a representative model for the flavone series and evaluated spectrophotometrically for potential to coordinate Cu(II) and/or Zn(II) ions implicated in the metal-catalyzed free radical generation. A plausible mechanism for metal-chelation of the test compounds is presented. Furthermore, the most active compounds from each series against the test carbohydrate-hydrolyzing enzymes were selected and evaluated for their antigrowth effect on the human breast (MCF-7) and lung (A549) cancer cell lines and for cytotoxicity against the African Green Monkey kidney (Vero) cell line. The parent chalcone 2a and flavone derivatives 3a, 3c and 3e exhibited relatively high inhibitory activity against the MCF-7 cells with IC50 values of 4.12 ± 0.55, 8.50 ± 0.82, 5.10 ± 0.61 and 6.96 ± 0.66 μM, respectively. The chalcones 2a and 2c exhibited significant cytotoxicity against the A549 cells with IC50 values of 7.40 ± 0.67 and 9.68 ± 0.80 μM, respectively. Only flavone 3c exhibited relatively strong and comparable cytotoxicity against the MCF-7 and A549 cell lines with IC50 values of 6.96 ± 0.66 and 6.42 ± 0.79 μM, respectively. Both series of compounds exhibited strong activity against the MCF-7 and A549 cell lines compared to the analogous quercetin (IC50 = 35.40 ± 1.78 and 35.38 ± 1.78 μM, respectively) though moderate compared to nintedanib (IC50 = 0.53 ± 0.11 and 0.74 ± 0.15 μM, respectively). The test compounds generally exhibited reduced cytotoxicity against the Vero cells compared to this anticancer drug. Molecular docking revealed strong alignment of the test compounds with the enzyme backbone to engage in hydrogen bonding interaction/s and hydrophobic contacts with the residues in the active sites of α-glucosidase and α-amylase. The test compounds possess favorable drug-likeness properties, supporting their potential as therapeutic candidates against T2DM.

INHIBITION OF ESTROGEN RECEPTOR ALPHA (ERα) BY BIOACTIVE COMPOUNDS FROM TERMINALIA ARJUNA (Roxb. ex DC.) Wight & Arn.: A MOLECULAR DOCKING STUDY

Estrogen receptor alpha (ERα) plays a critical role in breast cancer. Its overabundance can be driven by factors that stimulate estrogen hormone gene expression in living organisms. This, in turn, can lead to the development of several desirable properties by the cancer cells, impairing the maintenance of a regular mammary gland in females. Consequently, ERα offers a wide range of potential biochemical therapeutic targets for clinical research. Terminalia arjuna, a widely accepted medicinal plant in traditional medicine, has shown promise in treating various critical diseases. Our previous studies using swissADME identified 20 bioactive compounds in T. arjuna with favorable pharmacokinetic properties. This study aimed to evaluate the potential of these bioactive compounds against ERα (PDB ID: 3ERT) using molecular docking studies with Autodock 4.2.6. The docking results revealed high binding affinities for the designed compounds, ranging from -3.1 to -9.4 kcal/mol. These findings suggest that T. arjuna derived compounds could be significant for the development of novel and improved anti-breast cancer agents.

7787 Primary Aldosteronism Medical Treatment Outcomes (PAMO): An International Consensus on Outcome Assessment Criteria and Multicentre Study

Abstract Disclosure: J. Yang: None. J. Burrello: None. J. Goi: None. T.A. Williams: None. W.F. Young: Consulting Fee; Self; Arena Pharmaceuticals, Inc.. Other; Self; Crinetics Pharmaceuticals. P.J. Fuller: None. P. Investigators: None. Primary aldosteronism (PA) is the most common endocrine cause of hypertension. Mineralocorticoid receptor antagonists, at adequate doses, lower blood pressure and cardiovascular risk in patients with PA. However, there is a lack of a standardized approach to medical therapy and subsequent outcomes assessment. We aimed to: 1) establish criteria for assessing outcomes of targeted medical treatment of PA, and 2) evaluate outcomes using these criteria across an international cohort. A panel of 31 PA experts from 31 centres across four continents used the Delphi method, including three rounds of online questionnaires, to reach consensus. Clinical data were then collected from consecutive patients with PA who started targeted medical treatment between 2016 and 2021 at the participating centres. Consensus was reached for six outcomes, including complete, partial, or absent biochemical response, and complete, partial, or absent clinical response. “Complete biochemical response” was defined as “correction of hypokalemia without potassium supplementation, and normalization of plasma renin activity or concentration”; while “absent biochemical response” was defined as “persistent hypokalemia and/or persistently suppressed or low plasma renin activity or concentration without change from baseline”. “Complete clinical response” was defined as “normal blood pressure without additional antihypertensive medications”; while “absent clinical response” was defined as “the same or increased blood pressure compared to before targeted treatment with same or more antihypertensive medications, not counting targeted therapy”. Of 1483 patients (mean age 52y, 48% female) from 28 centers, 232 (16%) had a complete clinical response, and 719 (49%) had a complete biochemical response at the most recent follow-up (median 29 months after treatment initiation). Compared to those with partial or absent clinical response, patients with complete clinical response had a significantly higher proportion of women and lower proportion with diabetes, stroke or left ventricular hypertrophy. They also had a significantly shorter duration of hypertension, lower BMI, higher eGFR and lower defined daily dose of antihypertensives at baseline. In contrast, these differences were not observed in those with a complete, partial or absent biochemical response. The defined daily dose of targeted therapy did not differ across the various response categories. In conclusion, we have established an international expert consensus for standardized criteria to evaluate the response to targeted medical treatment of PA, facilitate comparisons of outcome data and guide clinicians who manage PA. In a retrospective international cohort, less than half of medically-treated patients reached the established biochemical treatment targets. Further work is needed to increase the rate of complete response. Presentation: 6/3/2024

Statistical estimation theory detection limits for label-free imaging.

The emergence of label-free microscopy techniques has significantly improved our ability to precisely characterize biochemical targets, enabling non-invasive visualization of cellular organelles and tissue organization. However, understanding each label-free method with respect to the specific benefits, drawbacks, and varied sensitivities under measurement conditions across different types of specimens remains a challenge. We link all of these disparate label-free optical interactions together and compare the detection sensitivity within the framework of statistical estimation theory. To achieve this goal, we introduce a comprehensive unified framework for evaluating the bounds for signal detection with label-free microscopy methods, including second-harmonic generation, third-harmonic generation, coherent anti-Stokes Raman scattering, coherent Stokes Raman scattering, stimulated Raman loss, stimulated Raman gain, stimulated emission, impulsive stimulated Raman scattering, transient absorption, and photothermal effect. A general model for signal generation induced by optical scattering is developed. Based on this model, the information obtained is quantitatively analyzed using Fisher information, and the fundamental constraints on estimation precision are evaluated through the Cramér-Rao lower bound, offering guidance for optimal experimental design and interpretation. We provide valuable insights for researchers seeking to leverage label-free techniques for non-invasive imaging applications for biomedical research and clinical practice.

Ultrasensitive optical fiber SPR sensor enhanced by Au-NPs-film modified with functionalized CQDs for label-free detecting cobalt (II) ion

BackgroundCobalt, an essential trace element, is vital for maintaining human nervous system function, aiding in DNA synthesis, and contributing to red blood cell production. It is helpful for disease diagnosis and treatment plan evaluation by precisely monitoring its concentration changes in the human body. Despite extensive efforts made, due to its ultra-low concentration, the current limit of detection (LOD) as reported is still inadequate and cannot be satisfied with the precise clinical applications. Therefore, it is crucial to develop novel label-free sensors with high sensitivity and excellent selectivity for detecting trace amounts of Co2+. ResultsHere, an ultrasensitive optical fiber SPR sensor was designed and fabricated for label-free detection of Co2+ with ultra-low concentration. It is achieved by modifying the carboxyl-functionalized CQDs on the AuNPs/Au film-coated hetero-core fiber, which can specifically capture the Co2+, leading to changes in the fiber's surface refractive index (RI) and subsequent SPR wavelength shifts in the transmission spectrum. Both the Au film and AuNPs on the fiber are modified with CQDs, leveraging their large surface area to enhance the number of active sites and probes. The sensor exhibits an ultra-high sensitivity of approximately 6.67 × 1019 nm/M, and the LOD is obtained as low as 5.36 × 10−20 M which is several orders of magnitude lower compared to other conventional methods. It is also experimentally demonstrated that the sensor possesses excellent specificity, stability, and repeatability, which may be adapted for detecting real clinical samples. SignificanceThe CQDs-functionalized optical fiber SPR sensor exhibits substantial potential for precisely detecting Co2+ of trace amounts, which is especially vital for scarce clinical samples. Additionally, the sensing platform with sample sensor fabrication and measurement configuration introduces a novel, highly sensitive approach to biochemical analysis, particularly adapting for applications involving the detection of trace targets, which could also be employed to detect various biochemical targets by facile modification of CQDs with specific groups or biomolecules.

Causal relationship between prostatic diseases and prostate cancer: a mendelian randomization study

BackgroundAlthough it is thought that prostatitis or benign prostatic hyperplasia (BPH) is related to prostate cancer (PCa), the underlying causal effects of these diseases are unclear.MethodsWe assessed the causal relationship between prostatitis or BPH and PCa using a two-sample Mendelian randomization (MR) approach. The data utilized in this study were sourced from genome-wide association study. The association of genetic variants from cohorts of prostatitis or BPH and PCa patients was determined using inverse-variance weighted and MR Egger regression techniques. The direction of chance was determined using independent genetic variants with genome-wide significance (P < 5 × 10–6). The accuracy of the results was confirmed using sensitivity analyses.ResultsMR analysis showed that BPH had a significant causal effect on PCa (Odds Ratio = 1.209, 95% Confidence Interval: 0.098–0.281, P = 5.079 × 10− 5) while prostatitis had no significant causal effect on PCa (P > 0.05). Additionally, the pleiotropic test and leave-one-out analysis showed the two-sample MR analyses were valid and reliable.ConclusionsThis MR study supports that BPH has a positive causal effect on PCa, while genetically predicted prostatitis has no causal effect on PCa. Nonetheless, further studies should explore the underlying biochemical mechanism and potential therapeutic targets for the prevention of these diseases.

IGF-I assay methods and biologic variability: evaluation of acromegaly treatment response.

Serum insulin-like growth factor (IGF-I) is the primary biochemical measure of disease activity in patients with acromegaly, and the 2014 Endocrine Society guidelines recommended normal age-adjusted serum IGF-I as the biochemical target of treatment. However, quantification and interpretation of IGF-I levels are subject to limitations that may affect therapeutic decisions. Techniques for measuring IGF-I have evolved greatly over the past 40 years and continue to do so. Results can vary substantially for different assays, procedures, and laboratories. For any assay, the interpretation of IGF-I values requires robust reference ranges. Using currently available large normative databases, the upper limit of normal (ULN) for IGF-I in middle-aged and elderly individuals is lower than historical reference ranges. Thus, the goal of achieving IGF-I < 1× ULN is more demanding than in the past, and some patients with acromegaly who were classified as "normal" (IGF-I < 1× ULN) in previous studies would be reclassified as above the ULN based on newer normative data. In addition, substantial intra-individual, week-to-week variation in serum IGF-I levels (unrelated to assay performance) has been observed. With changes over time in the measurement of IGF-I and the advent of updated reference ranges derived from large normative databases, it is difficult to justify rigid adherence to the goal of maintaining IGF-I below the ULN for all patients with acromegaly. Instead, symptoms, comorbidities, and quality of life should be considered, along with growth hormone and IGF-I levels, when evaluating the need for further treatment.

In Vivo Evaluation of 68Ga-Labeled NOTA-EGFRvIII Aptamer in EGFRvIII-Positive Glioblastoma Xenografted Model

EGFRvIII is expressed only in tumor cells and strongly in glioblastoma and is considered a promising target in cancer diagnosis and therapy. Aptamers are synthetic single-stranded oligonucleotides that bind to biochemical target molecules with high binding affinity and specificity. This study examined the potential of the 68Ga-NOTA-EGFRvIII aptamer as a nuclear imaging probe for visualizing EGFRvIII-expressing glioblastoma by positron emission tomography (PET). EGFRvIII aptamer was selected using the SELEX technology, and flow cytometry and fluorescence microscopy verified the high binding affinity to EGFRvIII positive U87MG vIII 4.12 glioma cells but not to EGFRvIII negative U87MG cells. The EGFRvIII aptamer was conjugated with a chelator (1,4,7-triazanonane-1,4,7-triyl)triacetic acid (NOTA) for 68Ga-labeling. The 68Ga-NOTA-EGFRvIII aptamer was prepared using the preconcentration-based labeling method with a high radiolabeling yield at room temperature. Ex vivo biodistribution analyses confirmed the significantly higher tumor uptake of the 68Ga-NOTA-EGFRvIII aptamer in EGFRvIII-expressing xenograft tumors than that in EGFRvIII negative tumors, confirming the specific tumor uptake of the 68Ga-NOTA-EGFRvIII aptamer in vivo. PET imaging studies revealed a high retention rate of the 68Ga-NOTA-EGFRvIII aptamer in U87MG vIII 4.12 tumors but only low uptake levels in U87-MG tumors, suggesting that the 68Ga-NOTA-EGFRvIII aptamer may be used as a PET imaging agent for EGFRvIII-expressing glioblastoma.

Multi-analyte responsive luminescent probes for the detection of biochemical targets in cell models

Luminescence has found wide application in biology, biotechnology and medicine. Particularly, fluorescent and bioluminescent probes allow visualization of molecular targets at the cellular level and even macromolecules or single small-molecule analytes. Among the most reliable tools for visualization of molecular targets are so-called responsive probes, which change the intensity and colour of the emitted signal after interaction with a molecular target. The majority of such probes allow detection of a single analyte. Meanwhile, most of the processes in the human body involve multiple elements. To better understand these mechanisms, it is possible to use several responsive probes simultaneously. However, this poses a risk of their different uptake by cells or different metabolism. In order to provide a more reliable response, so-called multi-analyte and multi-responsive probes are being developed. Examples of such probes will be discussed in the article, divided based on their response mechanism and detected changes within the cell.

Oral Estrogen Receptor PROTAC Vepdegestrant (ARV-471) Is Highly Efficacious as Monotherapy and in Combination with CDK4/6 or PI3K/mTOR Pathway Inhibitors in Preclinical ER+ Breast Cancer Models.

Estrogen receptor (ER) alpha signaling is a known driver of ER-positive (ER+)/human epidermal growth factor receptor 2 negative (HER2-) breast cancer. Combining endocrine therapy (ET) such as fulvestrant with CDK4/6, mTOR, or PI3K inhibitors has become a central strategy in the treatment of ER+ advanced breast cancer. However, suboptimal ER inhibition and resistance resulting from the ESR1 mutation dictates that new therapies are needed. A medicinal chemistry campaign identified vepdegestrant (ARV-471), a selective, orally bioavailable, and potent small molecule PROteolysis-TArgeting Chimera (PROTAC) degrader of ER. We used biochemical and intracellular target engagement assays to demonstrate the mechanism of action of vepdegestrant, and ESR1 wild-type (WT) and mutant ER+ preclinical breast cancer models to demonstrate ER degradation-mediated tumor growth inhibition (TGI). Vepdegestrant induced ≥90% degradation of wild-type and mutant ER, inhibited ER-dependent breast cancer cell line proliferation in vitro, and achieved substantial TGI (87%-123%) in MCF7 orthotopic xenograft models, better than those of the ET agent fulvestrant (31%-80% TGI). In the hormone independent (HI) mutant ER Y537S patient-derived xenograft (PDX) breast cancer model ST941/HI, vepdegestrant achieved tumor regression and was similarly efficacious in the ST941/HI/PBR palbociclib-resistant model (102% TGI). Vepdegestrant-induced robust tumor regressions in combination with each of the CDK4/6 inhibitors palbociclib, abemaciclib, and ribociclib; the mTOR inhibitor everolimus; and the PI3K inhibitors alpelisib and inavolisib. Vepdegestrant achieved greater ER degradation in vivo compared with fulvestrant, which correlated with improved TGI, suggesting vepdegestrant could be a more effective backbone ET for patients with ER+/HER2- breast cancer.

Synthesis, structure of the N-(Alkyl/Arylsulfonyl) substituted 5-(Bromo/Iodo)-3-methylindazoles and bioactivity screening against some of the biochemical targets linked to type 2 diabetes mellitus

The indazole scaffold and sulfonamide moiety have evoked high interest in medicinal chemistry especially in the context of antidiabetic drug development. A series of the C-5 unsubstituted (2a–f), 5‑bromo (2g–l) and 5-iodo (2m–r) substituted 1-(alkyl/arylsulfonyl)-3-methylindazoles were prepared, characterized and then evaluated through enzymatic assay in vitro for anti-α-glucosidase activity and for potential free radical scavenging properties. Compounds 2b, 2d, 2f–h, 2k, 2p and 2q exhibited strong activity against α-glucosidase compared to acarbose (IC50 = 1.30 ± 0.02 µM) with IC50 values in the range of 0.72 ± 0.03 – 1.20 ± 0.01 µM. These compounds exhibited increased or strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) and/or nitric oxide (NO) free radical scavenging activities. The compounds were, in turn, evaluated through enzymatic assays in vitro for the potential to inhibit or activate superoxide dismutase (SOD), lipoxygenase-15 (LOX-15) and/or vascular endothelial growth factor receptor-2 (VEGFR-2) activities. An in vitro cell-based antioxidant activity assay involving lipopolysaccharide induced reactive oxygen species (ROS) production in the human breast (MCF-7) cancer cell line revealed that compounds 2d, 2f and 2h exhibit moderate activity (35.5 %, 47 % and 28 %, respectively) compared to staurosporine (24 %). Compounds 2d, 2f and 2h significantly reduced the ROS level in the human embryonic kidney (HEK293-T) cells (65.5 %, 58 % and 48 %, respectively) compared to staurosporine (34 %). The planar benzo-fused heterocyclic scaffold and the distorted tetrahedral geometry of the N-sulfonyl moiety are predicted to facilitate hydrophobic and/or hydrogen bonding interactions to stabilize protein-ligand interaction in the receptors. The ADME analysis predicted that the test compounds have drug-like properties, a safe toxicity profile in silico without any carcinogenic and mutagenicity side effect. Molecular modelling was also used to attempt to rationalize the biological activity of these compounds and to correlate the results of in vitro and in silico studies.

Role of Colchicine in Cardiovascular Disorders.

Inflammation has played a pivotal role in atherosclerosis and other cardiovascular disorders, prompting the exploration of anti-inflammatory therapies to improve cardiovascular outcomes. Colchicine, a well-established agent in conditions such as gout and familial Mediterranean fever, has emerged as a promising novel anti-inflammatory agent in the realm of cardiovascular diseases. Its ability to target both traditional risk factors and residual inflammatory risk marks a significant advancement in cardiovascular prevention strategies, indicating a new era in cardiovascular care. Landmark trials have supported the efficacy and safety of low-dose colchicine in reducing major adverse cardiovascular events when combined with standard therapies. In addition, its endorsement by major cardiovascular societies underscores its significance as the first targeted anti-inflammatory therapy for cardiovascular disease. However, careful monitoring for drug interactions and adverse effects, particularly on kidney and liver function, is essential for safe use. In this review, we aim to comprehensively summarize the mechanisms of action of colchicine, its molecular and biochemical targets in various cardiovascular conditions, and its pharmacokinetics, and delve deeply into the existing evidence on its safety and efficacy in the treatment of cardiovascular disorders, including coronary artery disease, pericarditis, atrial fibrillation, and heart failure.

Biochemical bases of the organoprotective properties of metformin

Metformin is recognized as the "gold standard" for the treatment of type 2 diabetes mellitus (DM2). Its hypoglycemic properties are realized by reducing insulin resistance, contributing to the utilization of glucose in liver cells, muscles and adipose tissue; suppression of gluconeogenesis in the liver. Due to the wide range of biochemical targets of its pharmacological action, the scientific community actively discusses the use of metformin as an organoprotective drug. Now, the beneficial effect of metformin on the course of neuropathy, diseases of the cardiovascular system, normalization of the composition and metabolic activity of the intestinal microbiota, the course of metabolically associated fatty liver disease has already been proven, the nephroprotective effect has also been proven, and the study of the anti-oncogenic properties of metformin continues. The number of publications devoted to research on the positive effects of metformin on various organs and systems is actively increasing, and data on the identification of new effects are regularly published. The purpose of this literature review is to analyze the pleiotropic effects of metformin at the level of biochemical interactions for a more detailed understanding of the principles of their implementation.

CLINICAL CHARACTERISTICS OF PATIENTS WITH FIBROMUSCULAR DYSPLASIA WITH AND WITHOUT SPONTANEOUS CERVICAL ARTERY DISSECTION IN THE ARCADIA-POL STUDY

Objective: Fibromuscular dysplasia (FMD) is idiopathic, segmental, non-inflammatory, non-atherosclerotic disease affecting small and medium-sized arteries. FMD causes artery stenosis, although it may also lead to aneurysm, dissection, or arterial tortuosity. The aim of the study was to evaluate clinical characteristics of patients with spontaneous cervical artery dissection (SCeAD) and to assess prevalence of SCeAD among patients with FMD. Design and method: The ARCADIA-Pol is an ongoing study, initiated in 2015 at the Department of Hypertension, National Institute of Cardiology in Warsaw, Poland. Patients enrolled in the study met the inclusion criteria: presence or suspicion of FMD and/or radiologically documented episode of spontaneous dissection of any artery. Each patient underwent: clinical and 24h ambulatory blood pressure monitoring, biochemical tests, echocardiography, US Doppler of renal and carotid arteries and whole body angio-CT. We present data of 435 currently enrolled patients. Results: Among 435 patients included in the analysis SCeAD was confirmed in 77 subjects, most often in internal carotid arteries (68.8%). Mean age was 43,9 ± 9,6 years, majority of patients were women (64.9%). Hypertension (HT) was present in 59.7% patients, hypercholesterolemia in 44,2%. Among 77 subjects with SCeAD, the presence of typical FMD changes was confirmed in 35.1% cases. Patients with SCeAD and FMD were older and more often female as compared to those with SCeAD and without FMD. There were no differences in biochemical parameters or target organ damage parameters between SCeAD patients with and without FMD. Among 281 subjects with FMD, an episode of SCeAD was documented in 9.6% patients. Patients with FMD and SCeAD less often had HT. Women with FMD and SCeAD had more often a history of contraceptive pills use than those without SCeAD. No other differences were found between compared groups. Conclusions: FMD changes often coexist in patients with SCeAD (35.1%). In FMD patients complication of SCeAD occur relatively often (9.6%), which leads to conclusion that FMD is generalized arteriopathy. It is recommended to perform radiological screening in FMD patients to look for arterial complications in other vascular beds.

The Discovery of New Inhibitors of Insulin-Regulated Aminopeptidase by a High-Throughput Screening of 400,000 Drug-like Compounds.

With the ambition to identify novel chemical starting points that can be further optimized into small drug-like inhibitors of insulin-regulated aminopeptidase (IRAP) and serve as potential future cognitive enhancers in the clinic, we conducted an ultra-high-throughput screening campaign of a chemically diverse compound library of approximately 400,000 drug-like small molecules. Three biochemical and one biophysical assays were developed to enable large-scale screening and hit triaging. The screening funnel, designed to be compatible with high-density microplates, was established with two enzyme inhibition assays employing either fluorescent or absorbance readouts. As IRAP is a zinc-dependent enzyme, the remaining active compounds were further evaluated in the primary assay, albeit with the addition of zinc ions. Rescreening with zinc confirmed the inhibitory activity for most compounds, emphasizing a zinc-independent mechanism of action. Additionally, target engagement was confirmed using a complementary biophysical thermal shift assay where compounds causing positive/negative thermal shifts were considered genuine binders. Triaging based on biochemical activity, target engagement, and drug-likeness resulted in the selection of 50 qualified hits, of which the IC50 of 32 compounds was below 3.5 µM. Despite hydroxamic acid dominance, diverse chemotypes with biochemical activity and target engagement were discovered, including non-hydroxamic acid compounds. The most potent compound (QHL1) was resynthesized with a confirmed inhibitory IC50 of 320 nM. Amongst these compounds, 20 new compound structure classes were identified, providing many new starting points for the development of unique IRAP inhibitors. Detailed characterization and optimization of lead compounds, considering both hydroxamic acids and other diverse structures, are in progress for further exploration.