Bioorganic & Medicinal Chemistry最新文献

{"title":"Carboxylesterase 1d-mediated aglycone recognition is crucial for substrate processing and allosteric activation of endo-α-mannosidase in the endoplasmic reticulum","authors":"Akito Taira ,&nbsp;Taiki Kuribara ,&nbsp;Mitsuaki Hirose ,&nbsp;Kiichiro Totani","doi":"10.1016/j.bmc.2026.118547","DOIUrl":"10.1016/j.bmc.2026.118547","url":null,"abstract":"<div><div>Most nascent glycoproteins entering the endoplasmic reticulum (ER) undergo quality control <em>via</em> the calnexin/calreticulin (CNX/CRT) cycle, wherein Glc₁Man₉GlcNAc₂ (G1M9)-type glycans play a crucial role in monitoring protein folding. We have recently identified an <em>endo</em>-α-mannosidase activity within the ER, designated as ER-EM, which facilitates the release of misfolded glycoproteins from this cycle by converting G1M9-proteins into Man_8AGlcNAc₂ (M8A)-proteins in a single step. ER-EM appears to function as a complex comprising UDP-Glc:glycoprotein glucosyltransferase 1 (UGGT1), ERp57, and carboxylesterase 1D (Ces1d), although the role of Ces1d—primarily recognized for its involvement in lipid metabolism—in glycan-associated substrate recognition remains unclear. To elucidate the molecular basis of Ces1d-dependent recognition, we semi-synthesized a glycoprobe, <strong>G1M9-va-JW972</strong>, by conjugating the Ces1d-specific inhibitor JW972 to the aglycone of G1M9 using a linker <em>via</em> a click reaction. In the ER-EM reaction with this probe, <strong>M8A-va-piperidine</strong> was detected as an ER-EM product with the aglycone structural conversion <em>via</em> Ces1d-mediated hydrolysis of the JW972 moiety, demonstrating recognition of the substrate aglycone by the Ces1d component of ER-EM complex. Inhibition of the lipolysis site of Ces1d with WWL229 significantly reduced ER-EM activity, indicating that this site is essential for recognizing hydrophobic aglycones. Furthermore, the inactive substrate <strong>GlcMan-4MU</strong> was efficiently hydrolyzed in the presence of the Ces1d lipolysis site-specific inhibitor WWL229, demonstrating that the association of the hydrophobic compound WWL229 with the Ces1d lipolysis site contributes to allosteric activation of ER-EM. Our findings provide important insights into the functional regulation of ER-EM complex, a novel therapeutic target for protein misfolding diseases.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118547"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145922154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design, synthesis and biological evaluation of novel quinolinone derivatives as DprE1 inhibitors against Mycobacterium tuberculosis","authors":"Guoquan Wan ,&nbsp;Chao Gao ,&nbsp;Fei Teng ,&nbsp;Qifan Tang ,&nbsp;Jumei Zeng ,&nbsp;Luoting Yu","doi":"10.1016/j.bmc.2026.118551","DOIUrl":"10.1016/j.bmc.2026.118551","url":null,"abstract":"<div><div>Tuberculosis remains a global health crisis, exacerbated by the emergence of drug-resistant strains and limitations of current therapies. Aiming to the essential enzyme DprE1 in <em>Mycobacterium tuberculosis</em>, we designed and synthesized 34 novel quinolinone derivatives as non-covalent inhibitors. Among them, compound <strong>27</strong> demonstrated remarkable activity against Mtb H37Ra, with an MIC value of 0.2 ng/mL, and displayed low cytotoxicity against A549 cells. Further resistance profiling and molecular docking studies confirmed DprE1 as the primary target of the compounds, with the Y314H mutation being responsible for the development of resistance. These findings highlighted compound <strong>27</strong> as a suitable lead for developing novel and effective DprE1 inhibitors.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118551"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145916305","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design, synthesis, and biological evaluation of 2-amino-1,3,4-thiadiazoles as new potential EGFR inhibitors for treatment of cancer","authors":"Sachin M. Thorat ,&nbsp;Summon Koul ,&nbsp;Sachin Kote ,&nbsp;Ramakrishna G. Bhat","doi":"10.1016/j.bmc.2025.118527","DOIUrl":"10.1016/j.bmc.2025.118527","url":null,"abstract":"<div><div>1,3,4-Thiadiazoles are known to exhibit a wide array of biological activities such as diuretic, antifungal, anticonvulsant and anticancer properties. Since 1,3,4-thiadiazoles are structurally considered as bio-isosteres of pyrimidine, this structural framework along with its capability to bind to various biological targets has led to the development of many FDA approved drugs. The current work focuses on the identification of an initial hit from an existing library of thiadiazoles and its optimization to a potential lead molecule. The compounds were evaluated for in-vitro cytotoxicity in four human cancer cell lines namely HepG2, MCF7, HCT116 and A549 as well as in Vero cell lines. Four compounds <strong>11h</strong>–<strong>11k</strong> showed potency comparable with or better than the positive control Erlotinib and exemplary selectivity towards normal cells. These compounds were then screened to assess their inhibitory potential in wild type and mutated EGFR (T790M). Compound <strong>11j</strong> was found to be potent in these assays. Further computational approach driven SAR around this compound led to the identification of a more potent compound <strong>14h</strong> that showed sub-micromolar potency in wild type EGFR with much improved potency against T790M. Apoptotic effect of <strong>14h</strong> was investigated using annexin V-FITC/PI dual staining assay in which <strong>14h</strong> was found to trigger apoptotic pathways effectively with a significantly higher proportion of cells undergoing both early and late apoptotic events, underscoring its potential as a therapeutic agent targeting programmed cell death. Western blot analysis of <strong>14h</strong> indicates that it primarily impairs EGFR phosphorylation, supporting its potential as a targeted modulator of EGFR signalling.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118527"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145831748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis and biological evaluation of novel C1 or C3 amino derivatives of A-ring 1,2,3-trisubstituted 19-nor-vitamin D3","authors":"Tania González-García ,&nbsp;Lieve Verlinden ,&nbsp;Stefanie Doms ,&nbsp;Annemieke Verstuyf ,&nbsp;Susana Fernández ,&nbsp;Miguel Ferrero","doi":"10.1016/j.bmc.2025.118523","DOIUrl":"10.1016/j.bmc.2025.118523","url":null,"abstract":"<div><div>Two new analogs of 1α,25-dihydroxy-19-<em>nor</em>-vitamin D₃ with an additional hydroxyl group at position 2 and an amino group at C1 or C3 using quinic acid as starting substrate for the A-ring precursor are described. Key step was the deprotection of the <em>N</em>-benzyl group mediated by an efficient biocatalytic system of Laccase from <em>Trametes versicolor</em> together with TEMPO. The synthesized analogs did not exhibit any significant affinity for binding to either the VDR or hDBP when compared to the natural hormone and 1α,25-dihydroxy-19-<em>nor</em>-vitamin D₃. In terms of their antiproliferative activity in MCF-7 and HL-60 cells, the reported influence of the hydroxyl group at C1 on biological activity of vitamin D analogs is observed. The 1α,2α-dihydroxy-3α-amino derivative was most active than 1β-Amino-2β,25-dihydroxy-19-<em>nor</em>-vitamin D₃, exhibiting an antiproliferative activity in breast cancer MCF-7 cells at a concentration of 10⁻⁶ M comparable with 1α,25-(OH)₂-D₃ and 1α,25-(OH)₂–19-<em>nor</em>-D₃, but this compound was five times less potent than the natural hormone at the EC₅₀ concentration.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118523"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145779662","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discovery and in vivo evaluation of a fluorine-18 pro-drug tracer for imaging sphingosine-1-phosphate receptor 2 in the brain","authors":"Lin Qiu ,&nbsp;Wenjuan Zhou ,&nbsp;Hong Chen ,&nbsp;Hao Jiang ,&nbsp;Jiawen Lang ,&nbsp;Zhimin Xing ,&nbsp;Madison Heady ,&nbsp;Robert J. Gropler ,&nbsp;Caroline Guglielmetti ,&nbsp;Joel S. Perlmutter ,&nbsp;Zhude Tu","doi":"10.1016/j.bmc.2025.118536","DOIUrl":"10.1016/j.bmc.2025.118536","url":null,"abstract":"<div><div>Sphingosine-1-phosphate receptor 2 (S1PR2) plays an important role in demyelinating central nervous system (CNS) disease, such as multiple sclerosis (MS). To identify a suitable PET radiotracer for imaging S1PR2 in the brain, we synthesized thirteen new phthalazinyl-indazole containing S1PR2 ligands and determined their S1PR2 binding potencies <em>in vitro</em>. A racemic carboxylic acid and its enantiomeric isomers, (±)-<strong>5d</strong>, (−)-<strong>5d</strong>, (+)-<strong>5d</strong>, and 3-fluoropropoxyl analogues <strong>5e</strong>, and (±)-<strong>5f</strong> are potent with IC₅₀ values of 33.4, 13.4, 12.1, 27.7, and 16.4 nM for S1PR2, respectively. These compounds selectively bind S1PR2 over other S1PRs with IC₅₀s &gt;1000 nM. Three tracers (±)-[¹⁸F]<strong>5d</strong>, (−)-[¹⁸F]<strong>5d</strong>, (+)-[¹⁸F]<strong>5d</strong> were radiosynthesized with radiochemical yields of 38 %, 20 %, and 20 %, respectively, high radiochemical purity (&gt;95 %), and high molar activity (&gt;39 GBq/μmol, decay corrected to EOS). In addition, (±)-[¹⁸F]<strong>7a</strong>, a methyl ester pro-drug tracer of (±)-[¹⁸F]<strong>5d</strong>, was designed and synthesized with a radiochemical yield of 38 %, high radiochemical purity (&gt;95 %), and high molar activity (&gt;37 GBq/μmol, decay corrected to EOS). Our PET studies indicated (±)-[¹⁸F]<strong>5d</strong> had low mouse brain uptake with 0.35 %ID/g at 40–60 min post-injection, however, the pro-drug tracer (±)-[¹⁸F]<strong>7a</strong> had 11-fold higher brain uptake (4.10 %ID/g) than that of (±)-[¹⁸F]<strong>5d</strong> at 40–60 min post-injection. <em>Ex vivo</em> biodistribution revealed (±)-[¹⁸F]<strong>5d</strong> had consistently low brain uptake and no defluorination <em>in vivo</em> in SD rats. The radiometabolic analysis confirmed that (±)-[¹⁸F]<strong>7a</strong> was metabolized fast to form (±)-[¹⁸F]<strong>5d</strong> in rat plasma, and (±)-[¹⁸F]<strong>5d</strong> was ∼30 % of the total radioactivity at 45 min post-injection in rat brain. (±)-[¹⁸F]<strong>5d</strong> demonstrated relatively stable in nonhuman primate plasma <em>in vivo</em>. Furthermore, a PET brain study of (±)-[¹⁸F]<strong>7a</strong> showed that cuprizone-fed mice had reduced radioactive brain uptake compared with control mice, suggesting downregulation of S1PR2 expression in the brain of this demyelinating disease mouse model. Together, (±)-[¹⁸F]<strong>7a</strong> is a potential pro-drug PET radiotracer for imaging S1PR2 expression in the brain supporting further optimization of an S1PR2-specific radiotracer with appropriate <em>in vivo</em> radiolabeled metabolism kinetics.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118536"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145831780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel 7-Substituted-1,2,4-Triazolopyrimidines targeting Nav1.2 channels as low-neurotoxicity antiepileptic agents","authors":"Yuxi Zhou ,&nbsp;Weina Wang ,&nbsp;Cui Lv ,&nbsp;Jiaqi Wu ,&nbsp;Shuo Lv ,&nbsp;Luqian He ,&nbsp;Xinyu Zhao ,&nbsp;Xi Qin ,&nbsp;Binbin Tian ,&nbsp;Renhao Chen ,&nbsp;Chuanlong Guo ,&nbsp;Longjiang Huang ,&nbsp;Haibo Yu","doi":"10.1016/j.bmc.2026.118550","DOIUrl":"10.1016/j.bmc.2026.118550","url":null,"abstract":"<div><div>In pursuit of novel antiepileptic agents with improved safety profiles, a series of novel 7-substituted-[1,2,4]triazolo[1,5-<em>a</em>]pyrimidine derivatives were synthesized and evaluated for antiepileptic activity using the subcutaneous pentylenetetrazole (Sc-PTZ) and maximal electroshock seizure (MES) models <em>in vivo</em>. Compound <strong>6c</strong> showed significant antiepileptic effects in the MES model, with an ED₅₀ value of 13.70 mg/kg. Its TD₅₀ was greater than 261.85 mg/kg, yielding a protective index (PI) &gt;19.11. The antiepileptic efficacy of <strong>6c</strong> surpassed that of the reference drugs phenytoin and valproate. Notably, <strong>6c</strong> exhibited no neurotoxicity at its maximum soluble concentration, indicating a favorable safety profile. Docking and molecular dynamics (MD) simulations revealed stable binding of <strong>6c</strong> within the cavity of the Nav1.2 subunit (PDB: <span><span>6J8E</span><svg><path></path></svg></span>). Further electrophysiological evaluation demonstrated state-dependent blockade of Nav1.2 channels: at 10 μM, compound <strong>6c</strong> inhibited the inactivated state by 65 ± 18 % (<em>n</em> = 3), with dose-dependent inhibition characterized by an IC₅₀ value of 5.39 ± 0.81 μM (<em>n</em> = 5). These results suggest that compound <strong>6c</strong> is a promising lead candidate for the development of novel antiepileptic agents with high efficacy and low neurotoxicity. Preliminary <em>in vitro</em> assessment in rat liver microsomes indicated moderate metabolic stability for <strong>6c</strong>, supporting its potential for further development.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118550"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145922074","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discovery of valine thiazole-containing derivatives as potent reversers of P-glycoprotein–mediated multidrug resistance","authors":"Sihan Qiang ,&nbsp;Ke Wang ,&nbsp;Shanshan Wang ,&nbsp;Zongliang Liu ,&nbsp;Xin Yu ,&nbsp;Zuguang Yin ,&nbsp;Yuxin Feng ,&nbsp;Huaiyu Wang ,&nbsp;Huili Liu ,&nbsp;Hongbo Wang ,&nbsp;Yi Liu","doi":"10.1016/j.bmc.2025.118522","DOIUrl":"10.1016/j.bmc.2025.118522","url":null,"abstract":"<div><div>The development of multidrug resistance (MDR) to anticancer agents following chronic treatment remains a significant obstacle in cancer chemotherapy. Overexpression of the energy-dependent drug efflux transporter P-glycoprotein (P-gp) has been identified as a major contributor to MDR. Inspired by naturally occurring MDR-reversing cyclic peptides, a series of readily accessible valine thiazole-containing derivatives were designed and synthesized as effective reversal agents against P-gp–mediated MDR based on a hybridization strategy. Structure–activity relationship (SAR) analysis demonstrated that linear valine thiazole derivatives possess greater MDR reversal activity than their macrocyclic analogs. Compound <strong>17e</strong> displayed promising synergistic activity by significantly enhancing the therapeutic efficacy of paclitaxel (PTX) against KBV cells (IC₅₀ = 2.8 nM, reversal fold = 288.5) without inducing obvious toxicity. <strong>17e</strong> exhibited approximately 8-fold greater potency than verapamil (VRP), 2-fold greater potency than the third-generation P-gp inhibitor tariquidar (TAR) in KBV cells. Mechanistic studies confirmed that <strong>17e</strong> inhibits the efflux function of P-gp in a concentration-dependent manner without affecting its expression, thereby increasing intracellular chemotherapeutic drug accumulation and causing G₂/M phase arrests, which is associated with increased apoptosis. Taken together, the linear valine thiazole derivative <strong>17e</strong> represents a promising starting scaffold for developing novel P-gp inhibitors against MDR.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118522"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sequence-dependent RNA cleavage by the 8–17 DNAzyme: Identification of preferred tetranucleotide motifs","authors":"Satoko Nakajima ,&nbsp;Shuhei Miyakawa ,&nbsp;Shu Ohno ,&nbsp;Haruki Ikemoto ,&nbsp;Rikuto Maruyama ,&nbsp;Kosuke Chiba ,&nbsp;Ryota Suginaka ,&nbsp;Chihiro Oda ,&nbsp;Yuuya Kasahara ,&nbsp;Kaori Fukuzawa ,&nbsp;Satoshi Obika ,&nbsp;Takao Yamaguchi","doi":"10.1016/j.bmc.2025.118520","DOIUrl":"10.1016/j.bmc.2025.118520","url":null,"abstract":"<div><div>In recent years, deoxyribozymes (DNAzymes) capable of catalyzing RNA cleavage have attracted growing attention for their potential therapeutic applications. Among these, the 8–17 DNAzyme is one of the most extensively studied motifs, owing to its compact catalytic core and robust catalytic activity. Although it is well established that the 8–17 DNAzyme preferentially cleaves GG and AG (5′ → 3′) dinucleotide junctions within substrate RNA strands, the influence of flanking nucleotides on catalytic efficiency has not been systematically examined. Here, we comprehensively evaluated the tetranucleotide sequence preferences of the 8–17 DNAzyme. By assessing cleavage efficiencies across diverse tetranucleotide contexts in which the scissile phosphodiester bond lies between the second and third ribonucleosides, we identified three motifs—UGGG, UGGU, and UGGA—that were cleaved with markedly enhanced efficiency. Molecular dynamics and quantum chemical calculations suggested that stable formation of the noncanonical rG12·dA24 base pair in the UGGG motif facilitates a cleavage-competent structure, whereas this base pair is relatively disrupted in the GGGG motif, a less cleavable sequence. This sequence preference was further validated using 8–17 DNAzymes targeting mouse coagulation factor XI (<em>FXI</em>) mRNA. Collectively, these findings reveal a practical sequence preference of the 8–17 DNAzyme and provide a rational framework for designing highly active DNAzyme-based therapeutics.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118520"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145753894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"New pyrazolo[1,5-a]pyrimidine derivatives as potential CDK2 inhibitors and apoptosis-driven antiproliferative agents","authors":"Ahmed Abd El-Hamed El-Rahmany ,&nbsp;Nadia A. Khalil ,&nbsp;Mohamed S. Nafie ,&nbsp;Wagdy M. Eldehna ,&nbsp;Hatem A. Abdel-Aziz ,&nbsp;Marwa S.A. Hassan","doi":"10.1016/j.bmc.2025.118535","DOIUrl":"10.1016/j.bmc.2025.118535","url":null,"abstract":"<div><div>Five new series of pyrazolo[1,5-<em>a</em>]pyrimidine derivatives were synthesized as potential anti-cancer agents targeting the CDK2 enzyme. Their cytotoxic activity was evaluated against colon cancer (HCT-116) and breast cancer (MDA-MB-231) cell lines. Among the tested compounds, 7-(4-bromophenyl)-2-(methylthio)pyrazolo[1,5-<em>a</em>]pyrimidine-3‑carbonitrile (<strong>13</strong> <strong>g</strong>), 7-(2,4-dichlorophenyl)-2-(methylthio)pyrazolo[1,5-<em>a</em>]pyrimidine-3‑carbonitrile (<strong>13j</strong>), 7-[1-(4-fluorophenyl)-5-methyl-1<em>H</em>-1,2,3-triazol-4-yl)-2-(methylthio)pyrazolo[1,5-<em>a</em>] pyrimidine-3‑carbonitrile (<strong>21c</strong>), and 7-(6-bromo-2-oxo-2<em>H</em>-chromen-3-yl)-2-(methylthio)pyrazolo[1,5-<em>a</em>] pyrimidine-3‑carbonitrile (<strong>26b</strong>) exhibited strong growth inhibition in HCT-116 cells, comparable to the reference drug roscovitine. Compounds <strong>13</strong> <strong>g</strong> and <strong>21c</strong> were identified as the most potent candidates against HCT-116 cells, with IC₅₀ values of 0.45 μM and 0.09 μM, respectively, compared to roscovitine (IC₅₀ = 0.07 μM). Moreover, compounds <strong>13</strong> <strong>g</strong>, <strong>13j</strong>, <strong>21c</strong>, and <strong>26b</strong> also displayed low toxicity toward normal WI-38 fibroblast cells, indicating superior selectivity. CDK2 inhibition assay for the most potent compounds <strong>13</strong> <strong>g</strong>, <strong>13j</strong>, and <strong>21c</strong>, demonstrated promising IC₅₀ values ranging from 18 to 150 nM, compared to roscovitine (IC₅₀ = 140 nM). Further biological evaluation revealed that compound <strong>21c</strong> (IC₅₀ = 18 nM) triggered G1-phase cell cycle arrest and promoted more apoptosis than necrosis in HCT-116 cells (total apoptosis: 32.96 % compared to 0.63 % in the control). RT-PCR analysis indicated that apoptosis was mediated through both intrinsic and extrinsic pathways, accompanied by downregulation of the anti-apoptotic gene BCL-2 (0.7-fold change). Molecular docking studies supported these results, showing favorable interactions of compound <strong>21c</strong> within the CDK2 active site. Overall, compound <strong>21c</strong> emerges as a promising lead candidate for the development of selective CDK2 inhibitors exhibiting potent anti-cancer activity and low toxicity toward normal cells.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118535"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145808835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeting microbial β-CAs: bridging in silico screening with in vitro validation","authors":"Anna Rita Tondo ,&nbsp;Marialuigia Fantacuzzi ,&nbsp;Simone Carradori ,&nbsp;Ilaria D'Agostino ,&nbsp;Andrea Angeli ,&nbsp;Claudiu T. Supuran ,&nbsp;Clemente Capasso ,&nbsp;Nicola Gambacorta ,&nbsp;Luca Piemontese ,&nbsp;Antonio Laghezza ,&nbsp;Orazio Nicolotti ,&nbsp;Paolo Tortorella ,&nbsp;Mariangela Agamennone","doi":"10.1016/j.bmc.2025.118542","DOIUrl":"10.1016/j.bmc.2025.118542","url":null,"abstract":"<div><div>Antimicrobial resistance (AMR) is one of the world's most pressing health problems and requires immediate action from the scientific community. <em>Pseudomonas aeruginosa</em> is a particularly concerning bacterium due to its high level of resistance, especially among hospitalized and immunocompromised individuals. We recently investigated PsCA3, one of three <em>P. aeruginosa</em> β‑carbonic anhydrases (β-CAs), as a potential new pharmacological target for developing innovative antibacterial drugs. In this study, we applied a consensus structure-based virtual screening approach to select the most promising PsCA3 inhibitors from an in-house library of 607 small molecules. Twenty-one diverse compounds were selected and experimentally validated through enzyme inhibition assays, which assessed the ability of all ligands to block PsCA3 activity while sparing human α‑carbonic anhydrases (<em>h</em>CAs). Next, we focused on the benzoxazinone/dihydroquinolinone scaffold, testing fifteen additional analogues and expanding the set of assayed microbial β-CAs. Notably, we identified new, potent ligands based on unexplored scaffolds that can effectively target microbial β-CAs at micromolar/submicromolar concentrations with remarkable selectivity over human CAs.</div></div>","PeriodicalId":255,"journal":{"name":"Bioorganic & Medicinal Chemistry","volume":"134 ","pages":"Article 118542"},"PeriodicalIF":3.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145881201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}