The tyrosine kinase expressed in hepatocellular carcinoma (TEC) family comprises five nonreceptor tyrosine kinases─BTK, ITK, BMX, TXK, and TEC─with key roles in immune signaling. Although BTK and ITK have been extensively studied, selective inhibitors for TEC, TXK, and BMX remain scarce. Recently, we identified 7-azaindole-based covalent BMX inhibitors with potent inhibitory activity and robust cellular target engagement but limited selectivity across TEC family members, especially BTK. Here, we describe a new generation of BMX inhibitors designed to exploit subtle structural differences between BMX and BTK by incorporating diverse N-acylamino substituents at the azaindole 5-position or variations at the linker and warhead. Our compounds display subnanomolar BMX potency and improved selectivity over BTK and other TEC kinases. Key compound 11i showed strong cellular target engagement, good in vitro metabolic stability, a favorable kinome profile, and rapid covalent inactivation kinetics, positioning it among the best BMX chemical probes currently available.

Mitogen-activated protein kinase kinase 4 (MKK4), a MAP2 kinase that activates c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase, is a key kinase of the stress-activated protein kinase (SAPK)/mitogen-activated protein kinase (MAPK) signaling network. Inhibition of MKK4 represents a novel therapeutic approach by leveraging a rerouting mechanism within the signaling network, predominantly via MKK7 and JNK1, to modulate the regenerative capacity of hepatocytes. In this study, we describe the discovery of darizmetinib (HRX215), a first-in-class MKK4 inhibitor currently in clinical development. Darizmetinib was derived from a known BRaf inhibitor, and through extensive structure-activity relationship (SAR) studies, we successfully engineered potency and selectivity for MKK4 while eliminating the original BRaf on-target activity. Preclinical proof-of-concept studies, along with in vivo evaluations of different lead candidates, identified darizmetinib, demonstrating dose-dependent efficacy across various disease-relevant pharmacological models.

Purpose:The aim of this study was to discover and optimize a novel chemical scaffold capable of selectively inhibiting p38α, a kinase involved in inflammatory and neurodegenerative diseases. Despite decades of work, most p38α inhibitors have failed clinically due to limited selectivity, compensatory signaling, and safety issues. We sought to combine computational and experimental approaches to identify potent, drug-like, and selective inhibitors suitable for further development. Methods:A consensus virtual screening workflow (ESSENCE-Dock), integrating DiffDock, LeadFinder, and GNINA, was applied to the Eurofins-Villapharma compound library. The top hit guided similarity searching and clustering to identify related analogues for structure-activity relationship studies. Binding modes and substituent contributions were analyzed using molecular modeling and molecular dynamics simulations. Biochemical HTRF assays, ADME profiling, NanoBRET intracellular target engagement, and kinome-wide screening were used to evaluate potency, cellular activity, and selectivity. Results:Virtual screening identified a previously unreported 3,5-disubstituted dihydropyrazolo[1,5-a]pyrazinone scaffold as a potent p38α inhibitor (IC50 = 26 nM). Evaluation of related analogues yielded several compounds with sub-10 nM activity. Molecular dynamics simulations supported stable binding through interactions with MET109, ASP168, and LYS53. Selected compounds demonstrated high plasma stability, moderate solubility and permeability, strong intracellular target engagement (IC50 < 10 nM), and excellent selectivity across a 468-kinase panel. Conclusion:This study identifies and characterizes a novel and drug-like p38α inhibitor scaffold with potent biochemical activity, high kinome selectivity, and confirmed intracellular target engagement. The combined computational-experimental workflow provides a strong foundation for further optimization toward therapeutic candidates for inflammatory and neurodegenerative diseases.

Despite intensive efforts to improve early cancer detection, to date 30% of all solid tumors are still diagnosed at a metastasized and incurable stage. Molecular mechanisms underlying early metastasis are poorly understood, and thus far there are no strategies to suppress it. Here, using high-plex spatial protein profiling of human and murine tumours as well as functional studies in mouse models, we show that intratumoral microenvironmental stress factors induce senescence of apoptosis resistant tumor cells, resulting in a tumor-interlacing senescence matrix. Single cell RNA sequencing analyses revealed that matrix building cells, designated as stress-induced senescent tumor cells (SITC), harbour a distinct secretory phenotype, which fails to induce a full paracrine senescence phenotype in adjacent cancer cells but instead induces a hybrid senescence/invasion phenotype characterized by intermediate p16 levels and the upregulation of gene sets known to increase invasion and metastasis. Frequency of SITC in human therapy-naïve tumours was found to correlate with their metastatic stage at diagnosis and pharmacological or genetic depletion of SITC markedly reduced metastasis in orthotopic mouse models of colorectal cancer and intrahepatic cholangiocarcinoma. Our data harbours important translational potential, as recurrent pharmacological senolytic or senomorphic treatments hold the promise to eradicate SITC from occult tumors and this way prevent their early metastatic spread, allowing to diagnose more cancers at earlier and potentially curative stages. Clinical trials to address this hypothesis are warranted. Omelyan Trompak, Svenja A. Schütte, Jorge Abreu Macedo, Sophia Scheuermann, Zexi Hu, Igor Minia, Artür Manukyan, Thales Kronenberger, Marcello Zago, Rosanna Krebs, Alice Nuernbergk, Tae-Won Kang, Carlotta Schieler, Liudmyla Taranets, Angel M. Cuesta, Michael Forster, Kristin Bieber, Wolfgang Albrecht, Elke Rist, Can Yurttas, Daniel Dauch, Bettina Weigelin, Bence Sipos, Thomas Longerich, Stephan Singer, Markus W. Löffler, Manfred Claassen, Florian Wimmers, Markus Landthaler, Alfred Königsrainer, Saskia Biskup, Stefan Laufer, Christian M. Seitz, Lars Zender. Stress-induced intratumoral senescence matrix shifts cancer evolution towards metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6419.

The Hit to Lead (H2L) process is an integral part of contemporary drug discovery, encompassing the optimisation of validated Hit structures into Lead molecules. High quality leads build confidence, through activity and property profiles as well as preliminary biological data, which might include validating pharmacologic hypotheses along the way, indicating that further investment in the structure(s) and target would be worthwhile. Leads have line of sight to a development candidate and bring an understanding of what priorities Lead Optimisation should address. In this set of best practices, we detail the essential criteria that characterise a good lead, which include establishing SAR from analogues and assessing preliminary DMPK indicators, selectivity and early safety parameters. We highlight the importance of identifying liabilities of the lead series and demonstrating that each can be individually modulated whilst maintaining on target potency. We make the case for having physicochemical properties as critical optimisation parameters and how ligand efficiency metrics can enable this. Then we go over general tactics that can be used to convert hits into a lead series. These include essential steps that, when performed early, increase the chance of success such as deconstructive SAR, pharmacophore and bioactive conformation determination and scaffold optimisation. Finally, we suggest decision-making criteria to substantiate confidence in further investment or, as importantly, making a recommendation to cease further work on a series.

INTRODUCTION:Atypical chemokine receptor 3 (ACKR3) (formerly CXCR7) regulates various biological processes through its ligands and is closely associated with numerous diseases, including inflammation, cancer, cardiovascular diseases (CVDs), pain, and neurological disorders. Therefore, ACKR3 has emerged as a potential target for disease treatment. AREAS COVERED:This review summarizes the ACKR3 modulators published in patents from 2019 to 2024 using data from Google Patents, the European Patent Office, and the World Intellectual Property Organization's online databases. This includes information on their chemical structures, syntheses, activities, and developmental stages. EXPERT OPINION:ACKR3 agonists gained traction as a treatment for cardiovascular and pain conditions. WW-12, which was derived from the chemical modifications of conolidine, became a novel small-molecule pain modulator by activating ACKR3, which in turn boosted endogenous opioid peptides for the classical opioid receptors.ACKR3 antagonist ACT-1004-1239 from Idorsia Pharmaceuticals Ltd. has demonstrated the ability to treat cancer, acute lung injury/ARDS, and autoimmune diseases, including multiple sclerosis. The outcomes of these clinical trials will direct the development and indications of future ACKR3 modulators.

Lazertinib (YH25448) is a novel third-generation tyrosine kinase inhibitor (TKI) developed as a treatment for EGFR mutant non-small cell lung cancer. To better understand lazertinib inhibition at the molecular level, we determined crystal structures of lazertinib in complex with both WT and mutant EGFR and compared its binding mode to that of structurally-related EGFR TKIs. We observe that lazertinib binds with the novel pyrazole moiety involved in hydrogen bonds and van der Waals interactions consistent with drug potency and T790M mutant selectivity. Biochemical assays and cell studies confirm that lazertinib effectively targets EGFR(L858R/T790M) and to a lesser extent against HER2 as consistent with an improved toxicity profile. The molecular basis for lazertinib inhibition of EGFR reported here highlights new strategies for structure-guided design of tyrosine kinase inhibitors.

Pharmaceutical 3D printing (3DP) not only offers the possibility of dose personalization but also the co-administration of multiple active pharmaceutical ingredients (APIs) in one combination tablet. In this study, Theophylline (TPH) and Prednisolone (PSL) were printed as bi-tablets, which are single tablets with two distinct separate compartments. New findings show that the combination therapy of TPH with systemic corticosteroids shows a highly synergistic effect in the treatment of pulmonary diseases. For TPH, a drug with a narrow therapeutic window (NTW), precise sustained release requirements are mandatory, while PSL requires immediate drug release and is individually administered in doses specifically tied to the treatment progression. The study aims to understand the extent to which the combination of two tablet compartments influences the individual drug dissolution kinetics of the respective single compartments. Utilizing a full factorial statistical experimental design, various practically relevant doses were produced, investigated for their drug release, analyzed using different mathematical model fits, and compared with respective mono-tablets. The results show that the sustained drug release of TPH is not significantly influenced by the addition of a second compartment in relationship to respective doses. Individualization of bi-tablet doses while maintaining similar release profiles is possible with the given design setup, as release curves still show high similarity. In all tablet designs, PSL release occurred sufficiently fast, with the release rate correlating to the surface area-to-volume ratio (SA/V) as the main determining parameter.

The main protease (Mpro) of SARS-CoV-2 is a key drug target for the development of antiviral therapeutics. Here, we designed and synthesized a series of small-molecule peptidomimetics with various cysteine-reactive electrophiles. Several compounds were identified as potent SARS-CoV-2 Mpro inhibitors, including compounds 8n (IC50 = 0.0752 μM), 8p (IC50 = 0.0887 μM), 8r (IC50 = 0.0199 μM), 10a (IC50 = 0.0376 μM), 10c (IC50 = 0.0177 μM), and 10f (IC50 = 0.0130 μM). Most of them additionally inhibited cathepsin L and were also active against SARS-CoV-1 and MERS-CoV Mpro. In Calu-3 cells, several inhibitors, including 8r, 10a, and 10c, displayed high antiviral activity in the nanomolar range without showing cellular toxicity. The cocrystal structure of SARS-CoV-2 Mpro in complex with 8p revealed covalent binding to the enzyme's catalytic residue Cys145 and showed specific, unprecedented interactions within the substrate binding pocket. Compounds 10c and especially 8n were effective against a panel of naturally occurring nirmatrelvir-resistant mutants, particularly E166V, and showed metabolic stability and additional favorable pharmacokinetic properties, making it a suitable candidate for further preclinical development.

The title compound, C14H9FN2O, crystallizes in the monoclinic space group P21/c. The dihedral angles between the central isoxazole ring and the 4-fluorophenyl and pyridine rings are 32.64 (5) and 32.70 (7)°, respectively.

Gliomas are the most common malignant brain tumors in adults, characterized by a high proliferation and invasiveness. Treatment remains challenging due to their immunomodulatory properties, which promote immune evasion and tumor progression. Given the substantial consumption of yerba mate in the southern hemisphere, this study evaluated the effects of caffeinated and decaffeinated yerba mate infusion extracts (ECaf and EDCaf, respectively) on glioblastoma cell lines (U87 and U251). Both extracts reduced cell viability in a dose-dependent manner, with U87 being more sensitive. ECaf and EDCaf inhibited cell migration and adhesion, particularly in U87 cells. Real-time PCR showed a reduction in CD73 and MMP2 expression in U87, whereas U251 exhibited a slight increase in MMP2 expression. CD73 enzymatic activity was reduced in U87 cells by both extracts but remained unaffected in U251 cells. Overall, the results suggest that caffeine absence does not alter the bioactivity of yerba mate extracts, highlighting their potential to modulate glioblastoma cell behavior.

Colorectal cancer (CRC) constitutes the second leading cause of cancer-related death worldwide and advanced CRCs are resistant to targeted therapies, chemotherapies and immunotherapies. p38α (Mapk14) has been suggested as a therapeutic target in CRC; however, available p38α inhibitors only allow for insufficient target inhibition. Here we describe a unique class of p38α inhibitors with ultralong target residence times (designated ULTR-p38i) that robustly inhibit p38α downstream signaling and induce distinct biological phenotypes. ULTR-p38i monotherapy triggers an uncontrolled mitotic entry by activating Cdc25 and simultaneously blocking Wee1. Consequently, CRC cells undergo mitotic catastrophe, resulting in apoptosis or senescence. ULTR-p38i exhibit high selectivity, good pharmaco-kinetic properties and no measurable toxicity with strong therapeutic effects in patient-derived CRC organoids and syngeneic CRC mouse models. Conceptually, our study suggests ultralong-target-residence-time kinase inhibitors as an alternative to covalent inhibitors, which, because of the lack of cysteine residues, cannot be generated for many kinase cancer targets. Rudalska et al. describe a novel class of p38α inhibitors with increased target residence time. They explore the drugs’ specificity, pharmacokinetics and toxicity profile and show that they are efficacious in the context of colorectal cancer.

All JNK isoforms play a specific role in various diseases. The role of the JNK2 isoform has so far received little attention compared to its JNK1 and JNK3 counterparts with JNK3 being a potential target for neurodegenerative diseases and an inhibitor with JNK1 bias being currently investigated in clinical trials. Using an iterative, structure-guided optimization approach starting from a reported reversible binding aminopyrazole-derived scaffold, novel highly potent JNK2/3 selective inhibitors were generated ("ligand-first approach"). These reversible inhibitors were further transformed to covalent inhibitors by attaching an electrophilic warhead moiety, able to address a conserved cysteine side chain present in JNKs. Reversible and covalent inhibitors presented in this study show high JNK2/3 isoform selectivity and activity in cells. The covalently acting lead compound 56d shows good kinetic data with a kinact/KI (JNK2) = 38,200 M-1 s-1 as well as cellular isoform selectivity and a clean kinome profile.

Due to the arising of clinically relevant resistant EGFR-related phenotype through innovative mechanisms, mainly EGFRL858R/T790M, the emergence of novel molecules with dual or multi-target affinity has presented a promising alternative to overcoming these resistance mechanisms. This study aimed to evaluate synthetic acrylamide quinoxaline derivatives against NSCLC cell lines with different overexpressed EGFR mutations and compare their DMPK profile. The biological activity of LASSBio-1971 and LASSBio-1974 was assessed through cytotoxicity (MTT and Sulforhodamine B assays), apoptosis induction, EGFR inhibition, cell cycle analysis (flow cytometry), immunofluorescence microscopy, cell membrane permeability (PAMPA assay), and metabolic stability in rat liver microsomes. LASSBio-1971 exhibited promising EGFR inhibition with favorable in vitro pharmacokinetic (PK) properties, including high gastrointestinal and blood-brain barrier permeability. LASSBio-1974 demonstrated nonselective mechanism inhibiting EGFR and mitotic machinery leading to apoptosis and cell cycle arrest at different phases. LASSBio-1971 and LASSBio-1974 emerge as EGFR inhibitors with equipotent cytotoxic effects on human NSCLC lines and different in PK profile. Further studies should be conducted with LASSBio-1974 to prove and understand its antimicrotubule action.

ADVERTISEMENT RETURN TO ISSUEEditorialNEXTJournal of Medicinal Chemistry Collection: Drug Discovery in GermanyStefan LauferStefan LauferMore by Stefan Lauferhttps://orcid.org/0000-0001-6952-1486, Matthias GehringerMatthias GehringerMore by Matthias Gehringerhttps://orcid.org/0000-0001-7408-5084, and Alexander TitzAlexander TitzMore by Alexander Titzhttps://orcid.org/0000-0003-0163-3419Cite this: J. Med. Chem. 2024, 67, 4, 2237Publication Date (Web):February 2, 2024Publication History Received15 January 2024Published online2 February 2024Published inissue 22 February 2024https://pubs.acs.org/doi/10.1021/acs.jmedchem.4c00122https://doi.org/10.1021/acs.jmedchem.4c00122editorialACS PublicationsCopyright © Published 2024 by American Chemical Society. This publication is available under these Terms of Use. Request reuse permissions This publication is free to access through this site. Learn MoreArticle Views4288Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InRedditEmail PDF (844 KB) Get e-AlertscloseSUBJECTS:Biotechnology,Drug discovery,Medicinal chemistry,Pharmaceuticals,Quality management Get e-Alerts

Abstract A major limitation in the treatment of cancer is the limited therapeutic index of currently available chemotherapies or molecularly targeted therapies. Drug related toxicities often necessitate restricted drug dosing regimens, resulting in subtotal tumor remissions, which represent an important cause for tumor recurrences and the development of therapy resistance. Following up on previous work suggesting Aurora kinase A (AURKA) as a promising therapeutic target for the treatment of TP53 altered liver carcinomas (Dauch D. et al., Nat. Med., 2016) and TP53; RB1 altered small cell lung carcinomas (Gong X. et al., Cancer Discovery, 2019), we here report on the development and characterization of first in class kinase sparing AURKA ligands, which selectively kill TP53 deficient liver cancer cells and TP53; RB1 deficient small cell lung cancer cells by tethering AURKA to its binding partner TPX2 in mitotic cells. Small molecule mediated stabilization of AURKA/TPX2 complexes resulted in formation of multipolar spindles and subsequent cell death through mitotic catastrophe. Mechanistically, we were able to show that the hypersensitivity of TP53 deficient liver cancer cells and TP53; Rb1 deficient small cell lung cancer cells towards AURKA ligands is due to low expression levels of the antiapoptotic protein MCL1, which is necessary for survival of cancer cells in prolonged mitosis. Our novel AURKA ligands do not affect the kinase function of AURKA and thus, in contrast to conventional AURKA kinase inhibitors, can be administered continuously to mice without inducing toxicity related to inhibition of kinase related non-canonical (non-mitotic) functions of AURKA in normal cells or TP53 and RB1 wildtype cancer cells. Preclinical therapy studies in cell-line derived xenograft (CDX) tumor models of small cell lung cancer (SCLC) and orthotopic models of TP53 altered hepatocellular carcinoma and TP53;Rb1 altered SCLC demonstrated marked monotherapeutic efficacy of AURKA ligands. Conceptually, our herein provided data suggest that small molecules designed to stabilize protein-protein interactions represent powerful anti-cancer drugs with exceptional therapeutic window and thus open a venue towards the development of novel cancer therapies. Citation Format: Athina Anastasia Moschopoulou, Melanie Henning, Benedikt Wagner, Dirk Flötgen, Juliander Reiner, Yulia Skliarenko, Stefan Zwirner, Clemens Hinterleitner, Marco Seehawer, Luana dArtista, Elke Rist, Tatu Pantsar, Thales Kronenberger, Daniel Dauch, Corinna Gehring-Khav, Antti Poso, Liudmila Andreeva, Matthias Schwab, Michael Forster, Wolfgang Albrecht, Stefan Laufer, Lars Zender. Development and characterization of kinase sparing TPX2-tethering Aurora kinase A ligands for cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4478.