Integrin-linked kinase (ILK) is a pseudokinase that directly interacts with β-integrins and plays a pivotal role in regulating focal adhesion function. ILK has been implicated in the development of various diseases, part...Integrin-linked kinase (ILK) is a pseudokinase that directly interacts with β-integrins and plays a pivotal role in regulating focal adhesion function. ILK has been implicated in the development of various diseases, particularly cancer. However, currently, no validated ligands for ILK have been reported. Here, we describe the identification of 3-cyano-quinolines that potently bind to ILK (K = ∼250 nM), and crystallographic studies revealed a type I binding mode. A medicinal chemistry campaign exploring structure-activity relationships (SAR) using a robust parallel synthesis approach provided comprehensive SAR and identified regions amenable to modification. In addition, we demonstrated that the optimized 3-cyano-quinoline (DHP) modulates actin cytoskeletal dynamics. This work highlights the first validated ILK ligands and establishes a foundation for future translational efforts, such as the development of selective PROTACs targeting ILK for degradation by the ubiquitin system.
Given the therapeutic potential of ROCK2 in pulmonary fibrosis (PF) and the significant efficacy of our previously identified lead compound , we initiated an optimization campaign focused on addressing physicochemical li...Given the therapeutic potential of ROCK2 in pulmonary fibrosis (PF) and the significant efficacy of our previously identified lead compound , we initiated an optimization campaign focused on addressing physicochemical liabilities through disrupting molecular planarity. By implementing "sp-rich" and "magic methyl" strategies, two series of ROCK2 inhibitors featuring 4,5,6,7-tetrahydropyrazolo[1,5-]pyrazine or imidazo[1,2-]pyridazine scaffolds were developed. A dedicated evaluation discovered the optimal compound , which exhibited potent ROCK2 inhibition (IC = 0.0081 μM), enhanced kinetic solubility (77 μg/mL in water), improved metabolic stability ( = 5.0 h for p.o), favorable kinase selectivity, and an acceptable safety profile. Notably, effectively suppressed TGF-β1-induced fibroblast activation and migration, while significantly attenuating bleomycin-induced PF . Moreover, mechanistic studies revealed that the antifibrotic effects of were mediated through inhibition of the TGF-β/Smad and ROCK2/STAT3 signaling pathways. Overall, these findings validate the proposed design strategy and deliver a novel candidate for ROCK2-targeted therapy in PF.
Shiroodi R, Montavon TJ, Nelson BM
… +35 more, Randolph JT, O'Flynn DE, Iu L, Hutchins CW, Baikstis T, Izquierdo-Ferrer J, Smyth D, Krishnan N, Maneshi MM, Brown JW, Afanador GA, Jain R, Karunan Partha S, Bigelow L, Nocek B, Talaga L, Gopalakrishnan SM, Balakrishnan A, Sheehan MM, Kurschner VA, Jejurkar PC, Nacham O, Hu D, Yarilina A, Bradley NP, Chapman AM, Thakur A, Panchal SC, Sunose M, Zoi I, Dart MJ, Shotwell JB, Benekareddy M, Sadowski RN, Swensen AM
Herein, we report the discovery and optimization of a series of cluster of differentiation 38 (CD38) inhibitors derived from a virtual ligand screening (VLS) campaign. VLS identified imidazopyridazine hit , which was opt...Herein, we report the discovery and optimization of a series of cluster of differentiation 38 (CD38) inhibitors derived from a virtual ligand screening (VLS) campaign. VLS identified imidazopyridazine hit , which was optimized to a novel and potent CD38 inhibitor peripheral tool () with an excellent pharmacokinetic profile. A cocrystal structure in addition to biophysical and biochemical characterization of is consistent with uncompetitive inhibition of CD38 via the formation of covalent adduct . Further structure- and property-based modifications of afforded sulfuryl pyrazole () with improved CNS distribution properties. Brain-penetrant thienopyrimidine shows robust rodent pharmacokinetics and target engagement across skin, lung, liver, and brain, making it an excellent CD38 tool for the study of indications requiring engagement of CD38 in the brain.
Phosphoinositide 3-kinase δ (PI3Kδ) has emerged as a promising therapeutic target for inflammatory respiratory diseases. Herein, we report the design and synthesis of a novel series of 4-methylquinazoline derivatives as...Phosphoinositide 3-kinase δ (PI3Kδ) has emerged as a promising therapeutic target for inflammatory respiratory diseases. Herein, we report the design and synthesis of a novel series of 4-methylquinazoline derivatives as potent and highly selective PI3Kδ inhibitors for the treatment of acute lung injury (ALI). Guided by structure-based drug design optimization of a -PI3K inhibitor scaffold, the lead compound was identified, exhibiting single-digit nanomolar potency against PI3Kδ and exceptional selectivity over other class I PI3K isoforms. Mechanistically, compound effectively modulated immune responses by suppressing pro-inflammatory M1 macrophage polarization while promoting the anti-inflammatory M2 phenotype. Possessing favorable pharmacokinetic properties, compound was evaluated in murine models of LPS-induced ALI and septic lung injury, where it significantly attenuated pulmonary edema and inflammatory infiltration. Collectively, compound represents a promising preclinical candidate for the therapeutic intervention of ALI.
Zerihun M, Chakraborty S, Abita A
… +11 more, Dey S, Lopez I, Wainer Shlomo R, Cinthakunta Sridhar MK, Davis L, Ertracht O, Atar S, Shalev DE, De Zotti M, Haileselassie B, Qvit N
Mitochondrial fission protein 1 (Fis1) and mitochondrial dynamics protein of 51 kDa (Mid51) regulate stress-induced mitochondrial fragmentation implicated in cardiovascular disease. Using homologous sequence analysis and...Mitochondrial fission protein 1 (Fis1) and mitochondrial dynamics protein of 51 kDa (Mid51) regulate stress-induced mitochondrial fragmentation implicated in cardiovascular disease. Using homologous sequence analysis and structure-guided design, we identified a linear peptide inhibitor (CVP-240) targeting the Fis1/Mid51 protein-protein interaction (PPI) and optimized it into a macrocyclic derivative (CVP-764). Both compounds bind Mid51 with high affinity, selectively disrupt Fis1/Mid51 signaling over Drp1-dependent interactions, and exhibit nanomolar binding in fluorescence polarization assays using FAM-conjugated tracers. In H9c2 cardiomyocytes, CVP-240 and CVP-764 preserve mitochondrial membrane potential, reduce reactive oxygen species, maintain mitochondrial network integrity, and improve cell viability under stress. Macrocyclization enhances proteolytic and serum stability and confers intrinsic cell permeability without the need for a cell-penetrating sequence. In silico ADMET profiling and preliminary toxicity studies support a favorable safety profile, establishing CVP-764 as a promising lead for targeting pathological mitochondrial fission.
Yang J, Zhang LY, Ni XF
… +17 more, Bai MY, Wang X, Fu AQ, Chen MH, Zhang MY, Qiao N, Wang ZX, Li QQ, Cai S, Wang Y, Yao YS, Yu YC, Chen WD, Sun SL, Zhang Z, Li NG, Shi ZH
The efficacy of FLT3 inhibitors in acute myeloid leukemia (AML) is severely limited by resistance mutations, particularly the recalcitrant gatekeeper F691L and activation loop D835V/Y variants. Herein, through the struct...The efficacy of FLT3 inhibitors in acute myeloid leukemia (AML) is severely limited by resistance mutations, particularly the recalcitrant gatekeeper F691L and activation loop D835V/Y variants. Herein, through the structure-guided optimization of our previously reported lead , we identified , a highly potent type II FLT3 inhibitor capable of overcoming these recalcitrant mutants. exhibited single-digit nanomolar potency against Ba/F3-FLT3-ITD-F691L (IC = 9.09 nM) and subnanomolar activity against Ba/F3-FLT3-ITD-D835Y (IC = 4.64 nM) and Ba/F3-FLT3-ITD-D835V (IC = 0.76 nM). , induced profound tumor regression (TGI = 125%) in the MV4-11 xenograft model (10 mg/kg) and achieved robust tumor growth suppression (TGI = 68.6%) in the Ba/F3-FLT3-ITD-F691L model (50 mg/kg), where quizartinib was ineffective. Mechanistic studies confirmed that effectively blocked FLT3 signaling and induced apoptosis without observable toxicity. Collectively, represents a promising lead candidate for drug-resistant AML.
Only a handful of bacterial strains are responsible for the majority of clinical infections, with being the single largest contributor. infections can be very hard to confirm and localize. There is a clinical need for...Only a handful of bacterial strains are responsible for the majority of clinical infections, with being the single largest contributor. infections can be very hard to confirm and localize. There is a clinical need for improved imaging technologies to detect and characterize bacterial infections allowing for prompt initiation of treatment. We generated a molecule for positron emission tomography (PET) imaging of bacterial infection by linking two clinically used small molecules, Vancomycin (Vanco) and desferrioxamine B (DFO). This siderophore-antibiotic conjugate marries the high specificity of the antibiotic for Gram-positive bacteria with the receptor targeting and chelation capabilities of the siderophore. The conjugate can be labeled with Ga for detection by PET. We report here the scalable synthesis of the heterodimeric probe [Ga][Ga(DFO-PEG-Vanco)], the biochemical validation of its mechanism of action, its biodistribution, and the ability to image infection .
Smith JM, Barlaam B, Beattie D
… +21 more, Bradshaw L, Chan HM, Cooke SL, Cronin A, Cumming I, Dean E, Debreczeni JÉ, Barco Barrantes ID, Ferguson D, Gianni D, Grondine M, Lynch JT, McWilliams L, Moore S, Raubo P, Qu Y, Robb GR, Tan L, Urosevic J, Vazquez-Chantada M, Wang P
Inhibition of the arginine methyltransferase protein arginine methyltransferase 5 (PRMT5) has emerged as a key target for cancer therapy. Leveraging the MTAP synthetic lethality mechanism, MTA-cooperative PRMT5 inhibitor...Inhibition of the arginine methyltransferase protein arginine methyltransferase 5 (PRMT5) has emerged as a key target for cancer therapy. Leveraging the MTAP synthetic lethality mechanism, MTA-cooperative PRMT5 inhibitors are showing promising potential as precision cancer treatments with a high therapeutic index. Herein, we report our efforts to further optimize our previously reported tool compound ("AZ-PRMT5i-1") toward a clinical candidate-quality profile, by addressing key shortcomings of this compound─limited aqueous solubility, low hERG receptor activity, and an unfavorable predicted human dose. Exploration of the terminal lactam substitution group and the central aromatic group of the isindolinone scaffold provided the key structure-activity relationship insights to meet these goals. The highest quality compounds in this series were identified by the use of a dose-to-human (D2H) automated model. These efforts resulted in the identification of which shows the appropriate physicochemical properties, DMPK characteristics, and PRMT5-driven activity to be selected for progression into clinical studies.
Simultaneous targeting of glycolysis and oxidative phosphorylation (OXPHOS) is an effective strategy for overcoming the metabolic plasticity of pancreatic ductal adenocarcinoma (PDAC). In this study, we present compound...Simultaneous targeting of glycolysis and oxidative phosphorylation (OXPHOS) is an effective strategy for overcoming the metabolic plasticity of pancreatic ductal adenocarcinoma (PDAC). In this study, we present compound , a rationally designed, mitochondria-targeted small molecule that disrupts PDAC energy metabolism. Compound markedly inhibited PDAC cell glycolysis and mitochondrial function, as evidenced by the PDKs inhibition and the downregulation of OXPHOS-associated proteins, including SDHB and SIRT3, in vitro and in vivo. Mechanistically, induced ferroptotic cell death, accompanied by lipid peroxidation, redox imbalance, and mitochondrial dysfunction. Importantly, also elicited hallmarks of immunogenic cell death (ICD), including calreticulin exposure and HMGB1 release. In a syngeneic PANC02 model, suppressed tumor growth with minimal systemic toxicity and recapitulated the metabolic inhibition and ICD-associated phenotypes observed in vitro. These findings support that could be used as a dual metabolic inhibitor and ICD inducer in PDAC therapy.
Phillips DP, Alper PB, Cho CY
… +71 more, Borkin D, Han D, Kochanek SE, Vidal-Biggart A, Herath A, Gurjar J, Lu W, Mathison CJN, Nelson JM, Pei W, Yan S, Nguyen BN, Hoffman T, Okram B, Nguyen TN, Jiang S, Masick BT, Wang Z, Nikulin V, Yin H, Chen Y, Juarez J, Sarkisova Y, Jia Y, Zhou V, Liu G, Liu X, Lu M, Taylor BL, Huynh L, Wang Y, Pacoma R, Baaten J, Topolewski K, Wright C, Nguyen T, Knee DA, Liu Q, Liu J, Li J, Virata C, Briones S, DiDonato M, Bursulaya B, Jones DH, Witmer D, Chu C, Jin H, Richmond W, Westling L, Hollenbeck T, Chong A, Franey S, Groessl T, Shapiro M, Effenberger K, Arroyo K, Bretz A, Honda A, Brown J, Sanchez CC, Gordon WP, Matzen JT, Osterman N, Cotesta S, Brachmann SM, Wilcken R, Zecri F, Beyer KS, Molteni V, Haling JR
The RAS Switch-II pockets' discovery enabled targeting a challenging molecular site. Covalent KRAS inhibitors inspired efforts to find compounds for other KRAS mutations, notably KRAS and KRAS. A macrocyclization strateg...The RAS Switch-II pockets' discovery enabled targeting a challenging molecular site. Covalent KRAS inhibitors inspired efforts to find compounds for other KRAS mutations, notably KRAS and KRAS. A macrocyclization strategy led to the identification of an exceptionally potent lead compound . Highly optimized interactions in the pocket yielded strong affinity against KRAS and KRAS, potent inhibition of phospho-ERK in various KRAS mutant cell lines, and tumor regression in mouse xenograft models.
Myeloperoxidase (MPO)-mediated oxidative stress drives inflammatory tissue injury, yet converting this enzyme activity into a selective and sustained imaging readout remains chemically challenging. To address this limita...Myeloperoxidase (MPO)-mediated oxidative stress drives inflammatory tissue injury, yet converting this enzyme activity into a selective and sustained imaging readout remains chemically challenging. To address this limitation, we report Mn-TyrCDTA, a manganese chelate designed to couple kinetic inertness with MPO-triggered activation and retention mechanism. Replacement of a flexible EDTA backbone with a rigidified CDTA scaffold improved the kinetic inertness 3-fold under a Zn challenge (dissociation = 61.7 min). Incorporation of a tyramine-derived phenolic moiety enabled MPO/HO-mediated, one-electron oxidation and covalent protein anchoring, resulting in a 3.6-fold relaxivity enhancement and prolonged inflamed tissue retention. In rat models of acute pancreatitis, contrast enhancement correlated with tissue MPO activity ( = 0.83), enabling quantitative disease severity stratification. Complementary Ga-TyrCDTA PET studies demonstrated enzyme-dependent tracer accumulation, and MPO inhibition reduced the imaging signal by 85% (R = 0.98). These findings establish a rational design framework for the quantitative imaging of neutrophil-driven oxidative tissue injury.
Major depressive disorder (MDD) remains a global concern, with current treatments often showing delayed or limited efficacy. This research describes the development of novel estradiol-indole hybrids designed to inhibit t...Major depressive disorder (MDD) remains a global concern, with current treatments often showing delayed or limited efficacy. This research describes the development of novel estradiol-indole hybrids designed to inhibit the serotonin transporter (SERT) and activate the estrogen receptor (ER). Among 18 synthesized compounds, exhibited potent SERT inhibition (IC = 16.92 ± 1.71 nM), strong ER agonism (EC = 3.66 ± 0.55 nM, approximately 6-fold selectivity over ERα), and favorable oral absorption and brain penetration. Notably, reduced immobility in forced swim and tail suspension tests in chronic unpredictable mild stress (CUMS) mice and improved social interaction while reducing inactivity in chronic social defeat stress (CSDS) models. Consistent with coordinated involvement of ER signaling and SERT regulation, increased hippocampal serotonin levels and altered ER/SERT expression and downstream markers such as phosphorylated cAMP response element-binding protein (p-CREB) and brain-derived neurotrophic factor (BDNF). These results position as a promising dual-target antidepressant candidate with superior efficacy.
To overcome the proteolytic instability of oncolytic peptides, we developed a novel "MeLysine-stapling" strategy through intramolecular cross-linking of two α-methyl-substituted lysine residues. Applying this strategy to...To overcome the proteolytic instability of oncolytic peptides, we developed a novel "MeLysine-stapling" strategy through intramolecular cross-linking of two α-methyl-substituted lysine residues. Applying this strategy to the cobra-derived peptide yielded a series of stabilized analogues. The dual-modified lead candidate (MeLysine-stapling and -terminal acetylation), exhibited an exceptional serum half-life exceeding 48 h─a 32-fold increase over the parent peptide. demonstrated potent broad-spectrum cytotoxicity (geometric mean IC = 3.9 μM across nine cancer cell lines) and low toxicity to human erythrocytes and normal mammalian cells. Mechanistic investigations confirmed that acts via rapid, irreversible physical disruption of cancer cell membranes. In a syngeneic 4T1 xenograft model, intravenous administration of (5 mg/kg, every other day) inhibited tumor growth by 63% without systemic toxicity or exacerbated pathologies. Collectively, this study validates MeLysine-stapling as a robust strategy for engineering stable peptide therapeutics and positions as a promising preclinical candidate.
The integrated stress response (ISR) is a highly conserved cellular pathway triggered by a variety of insults, reducing protein synthesis and inducing ATF4, leading to broadly remodeling the cellular transcriptome and me...The integrated stress response (ISR) is a highly conserved cellular pathway triggered by a variety of insults, reducing protein synthesis and inducing ATF4, leading to broadly remodeling the cellular transcriptome and metabolome. ISRIB, , the first identified eIF2B activator, attenuates the ISR restoring protein synthesis, but its poor solubility limits absorption and advancement. To improve drug-like properties, we explored replacements for both the cyclohexyl core and side chains of ISRIB. This effort initially led to truncated analogue, 2BAct, , which demonstrated improved solubility relative to ; however, cardiovascular effects in higher species limited its progression into the clinic. Potent analogue was identified with significantly improved solubility vs but was still projected to have solubility-limited absorption. A prodrug campaign resulted in the identification of compound (fosigotifator), which exhibited significantly improved solubility and is currently being investigated in the clinic.
Human Phospholipase A Group V (hPLA-G5) is elevated in inflammatory conditions and promotes neutrophil and macrophage recruitment. Its enzymatic activity activates lipid receptors and cytosolic phospholipase A (cPLA), le...Human Phospholipase A Group V (hPLA-G5) is elevated in inflammatory conditions and promotes neutrophil and macrophage recruitment. Its enzymatic activity activates lipid receptors and cytosolic phospholipase A (cPLA), leading to arachidonic acid release and PGE production─key mediators of chronic inflammation. Thus, hPLA-G5 represents a promising therapeutic target for diseases with inflammatory pain, such as in endometriosis─a highly debilitating disease characterized by the ectopic growth of endometriotic cells in the abdominal cavity. High-throughput and fragment-based screening identified structurally related small molecule hits. Optimization of the physicochemical and pharmacokinetic properties of the HTS hit led to the identification of BAY-439, a potent and selective hPLA-G5 inhibitor with single-digit nanomolar activity. Accepted as a donated chemical probe by the Structural Genomics Consortium, both BAY-439 and the inactive negative probe BAY-163 are freely available as valuable pharmacological tools to investigate the role of hPLA-G5 both under physiological and pathological conditions.
Accurate antibody structural modeling is essential for therapeutic antibody development. However, even advanced artificial intelligence models such as AlphaFold2/3 struggle to precisely predict antibody structures due to...Accurate antibody structural modeling is essential for therapeutic antibody development. However, even advanced artificial intelligence models such as AlphaFold2/3 struggle to precisely predict antibody structures due to the high flexibility of the complementarity-determining regions. Structure refinement methods can therefore play a critical role in improving the quality of predicted antibody conformations. In this paper, we present ABRefine, an end-to-end antibody structure refinement framework based on an equivariant graph transformer with rigid-body constraints. ABRefine directly predicts atomic coordinate shifts from initial structures while enforcing physically meaningful rigid-body consistency, enabling rapid and accurate refinement without conformational sampling. To our knowledge, ABRefine is also the first dedicated refinement method for nanobodies. Comprehensive evaluations on multiple antibody and nanobody benchmarks show that ABRefine outperforms existing refinement methods in both accuracy and efficiency, and notably enhances the structural quality of AlphaFold3 predictions.
Unimolecular multiagonists integrating GLP-1R/GIPR, and increasingly GCGR agonism, have altered the landscape of peptide therapeutics for metabolic syndrome. This progress has shifted the field from optimizing individual...Unimolecular multiagonists integrating GLP-1R/GIPR, and increasingly GCGR agonism, have altered the landscape of peptide therapeutics for metabolic syndrome. This progress has shifted the field from optimizing individual ligands to engineering defined receptor selectivity (balance) and, in some cases, pathway-selective signaling (bias) within a single-peptide scaffold. Despite this momentum, tuning 'balance' and 'bias' remains difficult because small sequence changes can have receptor-dependent, nonintuitive effects, and meaningful retuning often requires wholesale scaffold redesign. Here, we implement a systematic strategy based on simple N-terminal chemical modifications across dual- and triagonist templates to decouple these variables. Because the peptide N-terminus is buried within the membrane-embedded transmembrane core of class B GPCRs, localized chemical edits at this site provide a sensitive lever for reweighting receptor activation. These minimal changes can alter signaling preference without loss of efficacy, expanding the design grammar and offer rapid tuning of potency, efficacy, and bias.
As part of a therapeutic approach to idiopathic pulmonary fibrosis (IPF) using inhaled ALK5 inhibitors, which enable targeted lung delivery while minimizing systemic side effects, this work describes the optimization pro...As part of a therapeutic approach to idiopathic pulmonary fibrosis (IPF) using inhaled ALK5 inhibitors, which enable targeted lung delivery while minimizing systemic side effects, this work describes the optimization process of a previously reported series featuring a 4,6-disubstituted pyridazine core. The medicinal chemistry exploration, aimed at increasing cellular potency while keeping physicochemical and ADME properties favorable for inhalation, was directed to the functionalization of the 3-position in the pyridazine core. An efficient SAR exploration, supported by a late-stage functionalization (LSF) approach, led to the identification of a small set of compounds worthy of characterization. Compound showed a persistent and lung-restricted target engagement in a pharmacodynamic model, which well-correlated with its solubility measured in simulated lung fluid (SLF). When tested in a mouse model of lung fibrosis, showed remarkable efficacy, thus representing an advanced lead candidate for the development of topical antifibrotic therapeutics.
Homozygous deletion of the methylthioadenosine phosphorylase () gene occurs in 10-15% of all human cancers and up to 50% of high-grade malignant gliomas, representing one of the largest opportunities for precision oncolo...Homozygous deletion of the methylthioadenosine phosphorylase () gene occurs in 10-15% of all human cancers and up to 50% of high-grade malignant gliomas, representing one of the largest opportunities for precision oncology. Loss of MTAP leads to the accumulation of 5'-methylthioadenosine (MTA), which sensitizes tumor cells to inhibition of protein arginine methyltransferase 5 (PRMT5). Herein we describe the discovery of , a potent and highly selective MTA-cooperative PRMT5 inhibitor that is brain penetrant in preclinical species and currently in Phase I/II clinical studies for the treatment of advanced or metastatic solid tumors with MTAP loss, with a focus on glioblastoma.