KRAS-G12D mutations are common drivers of pancreatic and colorectal cancers, yet effective targeted therapies remain limited. This study describes the design, synthesis, and biological evaluation of two novel KRAS-G12D inhibitors, GD-2 and GD-4. Both compounds exhibited strong antiproliferative activity in AGS and ASPC1 cancer cell lines, with IC₅。 values ranging from 0.2 to 1.8 µM. The protein binding assay also demonstrated high affinity for KRAS-G12D, with dissociation constants (Kd) of 146 nM for GD-2 and 3.18 nM for GD-4. Mechanistic investigations revealed that both compounds significantly reduced downstream, as evidenced by a clear decrease in phospho-ERK expression. Additionally, molecular dynamics simulations confirmed stable binding interactions within the KRAS-G12D pocket. Collectively, these findings identify GD-2 and GD-4 as promising therapeutic candidates for KRAS-G12D-driven cancers.

Oncogenic RAS mutations, which are common in human tumors and occur in about 30 % of cancer cases, present significant challenges to effective cancer treatment. Among the KRAS family, the KRAS-G12D mutation is a promising target for treating different types of cancer. Current approaches to inhibit the KRAS-G12D mutation have shown limited success, highlighting the urgent need for innovative therapies. In this study, we employed machine learning, followed by scaffold and core hopping fragmentation, to design, synthesize, and biologically test several 1-oxa-3,7-diazaspirodecane-2-one compounds, ultimately identifying two new KRAS-G12D inhibitors. Multiple in silico evaluations were performed to explore the potential of these inhibitors and to gain a deeper structural understanding of how these compounds bind within the KRAS-G12D active site. Additionally, protein binding assays and other biological tests demonstrated that these compounds exhibit a strong protein binding affinity (Kd of 28.29, 48.17, and 85.17 nM) and high selectivity for KRAS-G12D. Subsequent cellular assays further prioritized HDB-2 and HDB-3 as potent KRAS-G12D inhibitors, each showing nanomolar IC50 values. These results suggest that these compounds could become highly effective and selective anticancer agents for targeting KRAS-G12D-driven tumors.

Nuclear export protein 1, also known as XPO1, plays a crucial role in cellular homeostasis and assists in the nucleocytoplasmic transfer of ribonucleic acids (RNAs) and proteins. In addition, this nuclear export receptor is essential for the export of a variety of cargo molecules, such as proteins implicated in the immune response, tumor suppression, and cell cycle regulation. XPO1 has emerged as a promising target to disrupt the life cycles of multiple viruses and treat cancers. In our current work, we used a computational approach consisting of pharmacophore-assisted virtual screening complemented by molecular docking, molecular dynamics, and solvation-based free-energy studies to identify new inhibitors of the XPO1 protein. The identified compounds displayed highly stable RMSD plots, hydrogen bonding interactions, and relatively good binding affinities in both docking and free energy studies. These molecules were validated in vitro against SARS-CoV-2 and cancer cell lines. The study identified novel inhibitors of the XPO1 protein with both antiviral and anticancer activities.

The emergence of C797S-driven Osimertinib resistance represents a major hurdle in the treatment of non-small cell lung cancer (NSCLC). This study presents a novel series of proteolysis-targeting chimeras (PROTACs) designed to overcome resistance by potently and selectively degrading EGFR mutants, including the C797S variant. These next-generation EGFR PROTACs demonstrated significant anti-proliferative activity across a broad spectrum of EGFR mutations, achieving subnanomolar potency in cellular proliferation and protein degradation assays alike. Notably, these compounds effectively targeted not only C797S mutants but also other clinically relevant mutations such as Exon19Del and L858R/T790M variants. The lead candidate, HDBNJ3049, exhibited superior activity across all tested EGFR mutant cell lines, including potent growth inhibition of C797S-mutant Ba/F3, and H1975 cells (GI50 as low as 17 nM and 50 nM for L858R/T790M/C797S Ba/F3 and H1975, respectively), and robust degradation of Del19/T790M/C797S mutant EGFR (DC50: 13 nM, Dmax: 98%). In vivo pharmacokinetic and pharmacodynamic studies further validated the therapeutic potential of HDBNJ3049, establishing it as a highly promising lead candidate for overcoming osimertinib resistance in NSCLC and offering new hope for patients with limited treatment options. Mohammad Hassan Baig, Yun Seong Jo, Sagar Dattatraya Nale, Chang Joong Kim, Tae Hwan Park, Ju Han Bok, Dong Min Kim, Ji Min Park, Hye Mi Kim, Jae June Dong, Byoung Gon Moon. HDBNJ3049: A novel EGFR-targeting PROTAC to overcome C797S-mediated osimertinib resistance in NSCLC [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 401.

KRAS G12D mutations, occurring in approximately 30% of solid tumors, represent a significant therapeutic target. While selective KRAS G12C inhibitors have achieved clinical success, effectively targeting the more prevalent G12D mutation remains an unmet medical need. This study investigates the potential of proteolysis-targeting chimeras (PROTACs) as a promising avenue for KRAS G12D-driven cancers, potentially offering advantages over traditional inhibitors through catalytic protein degradation. Targeted protein degradation of mutant KRAS may yield greater efficacy than simply inhibiting the protein. This study focuses on the design, synthesis, and biological evaluation of novel KRAS G12D-targeting PROTAC degraders. Through rigorous optimization, we developed HDB-82, a highly potent and selective PROTAC compound targeting KRAS G12D. HDB-82 effectively induced degradation of the KRAS G12D protein, leading to significant antiproliferative activity against KRAS G12D-mutant cancer cells in vitro and in vivo. HDB-82 demonstrated potent anti-cancer activity against various KRAS G12D-mutant cell lines, achieving low nanomolar IC50 values. It effectively degraded the KRAS G12D protein in various KRAS G12D-expressing cell lines (DC50 value ranges from picomolar to sub-nanomolar range). Mechanistic studies revealed significant suppression of downstream MAPK and PI3K/AKT signaling pathways, accompanied by robust induction of apoptosis, and superior antiproliferative activity compared to existing inhibitors. Furthermore, HDB-82 exhibited favorable pharmacokinetic and pharmacodynamic properties in mice. Intravenous administration of HDB-82 (10 mg/kg, q.w.) resulted in substantial tumor growth inhibition across multiple KRAS G12D-positive tumor models. These findings establish HDB-82 as a promising therapeutic candidate for KRAS G12D-mutant cancers, warranting further preclinical investigation and potential clinical development. Mohammad Hassan Baig, Yun Seong Jo, Sagar Dattatraya Nale, Chang Joong Kim, Tae Hwan Park, Ju Han Bok, Dong Min Kim, Hye Mi Kim, Ji Min Park, Jae June Dong, Jae Hee Cho, Sung Ill Jang, Byoung Gon Moon. HDB-82: A promising PROTAC degrader for KRAS G12D mutant cancers [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 7010.

Pathogenic coronavirus, including COVID-19, threatens human health, and there have been strong demands for efficient therapeutics. Cordyceps militaris is a medicinal mushroom that has long been used for immune enhancement, anticancer, and antiviral effects. Therefore, the inhibitory potentials of constituents of C. militaris against COVID-19 were analyzed using various virtual screening analyses. Among ten constituents of C. militaris, cordycepin, the major component, and 3'-deoxyuridine and 2'-O-methyl-adenosine showed strong binding affinity to Mpro, a potential target for COVID-19 therapeutics. Considering the structure-activity relationship, nucleosides having deoxyribose and methoxyribose moiety are important for the affinity to Mpro. Cordycepin is also bound to Mpro mutants, and the binding mechanisms between cordycepin and Mpro were investigated further by MD simulation and MM/PBSA analysis. Principal component analysis also confirmed the conformational change of Mpro by cordycepin, which inhibits the function of Mpro. In vitro, the efficacy of cordycepin was measured using Vero cells infected with SARS-CoV-2, which showed excellent inhibition with an IC50 value of 29 μM. Conclusively, the constituents of C. militaris are expected to inhibit SARS-CoV-2 replication through binding to Mpro. Therefore, C. militaris can be an essential therapeutic for coronavirus through the synergistic effect of its constituents.

Neuroblastoma (NB) is the most common extracranial childhood cancer, caused by the improper differentiation of developing trunk neural crest cells (tNCC) in the sympathetic nervous system. The N6-methyladenosine (m6A) epitranscriptomic modification controls post-transcriptional gene expression but the mechanism by which the m6A methyltransferase complex METTL3/METTL14/WTAP is recruited to specific loci remains to be fully characterized. We explored whether the m6A epitranscriptome could fine-tune gene regulation in migrating/differentiating tNCC. We demonstrate that the m6A modification regulates the expression of HOX genes in tNCC, thereby contributing to their timely differentiation into sympathetic neurons. Furthermore, we show that posterior HOX genes are m6A modified in MYCN-amplified NB with reduced expression. In addition, we provide evidence that sustained overexpression of the MYCN oncogene in tNCC drives METTL3 recruitment to a specific subset of genes including posterior HOX genes creating an undifferentiated state. Moreover, METTL3 depletion/inhibition induces DNA damage and differentiation of MYCN overexpressing cells and increases vulnerability to chemotherapeutic drugs in MYCN-amplified patient-derived xenografts (PDX) in vivo, suggesting METTL3 inhibition could be a potential therapeutic approach for NB.

The nuclear export protein 1 (XPO1) mediates the nucleocytoplasmic transport of proteins and ribonucleic acids (RNAs) and plays a prominent role in maintaining cellular homeostasis. XPO1 has emerged as a promising therapeutic approach to interfere with the lifecycle of many viruses. In our earlier study, we proved the inhibition of XPO1 as a therapeutic strategy for managing SARS-COV-2 and its variants. In this study, we have utilized pharmacophore-assisted computational methods to identify prominent XPO1 inhibitors. After several layers of screening, a few molecules were shortlisted for further experimental validation on the in vitro SARS-CoV-2 cell infection model. It was observed that these compounds reduced spike positivity, suggesting inhibition of SARS-COV-2 infection. The outcome of this study could be considered further for developing novel antiviral therapeutic strategies against SARS-CoV-2.

Abstract The occurrence of C797S mutation in epidermal growth factor receptor (EGFR) is a leading mechanism of clinically acquired resistance to third-generation EGFR inhibitors, including Osimertinib. L858R/T790M/C797S and del19/T790M/C797S are commonly observed tertiary EGFR mutants identified in Osimertinib-resistant tumors. As of now, no clinically approved treatment exists that specifically targets these mutants. Here, we report the design and synthesis of a series of highly effective next-generation EGFR degraders effectively degrading EGFR C797S-containing triple mutants. Most compounds demonstrated antiproliferation activity in the subnanomolar range when tested on Ba/F3L858R/T790M/C797S and del19/T790M/C797S cells. Not only C797S but our designed degraders also degraded a wide range of EGFR mutants, including Exon19Del and L858R/T790M (DC50 <100nM). One representative Compound, HDBNJ2812, strongly degrades L858R/T790M/C797S and del19/T790M/C797S with DC50 of 34 nM (Dmax 88.5%) and 14 nM (Dmax 99.7%), respectively. This compound potently inhibits the proliferation of Ba/F3L858R/T790M/C797S (GI50 64 nM) and del19/T790M/C797S (GI50 40 nM). HDBNJ2812 demonstrated high inhibitory potential on HCC827 (del19) and H1975 (L858R/T790M) cell lines (GI50 18.9 and 85 nM, respectively). Furthermore, this degrader demonstrates weak cytotoxicity on non-mutant EGFR-expressing cells, such as A431, WI-26 (human lung fibroblast cells), and CHO-K1 (Chinese hamster ovary cells). Additionally, the in vivo PK/PD findings complement this compound's potential to be considered further. HDBNJ2812 may serve as a lead compound to render the greater therapeutic window for treating resistant non-small cell lung cancer patients with EGFR C797S mutants. Citation Format: Mohammad Hassan Baig, Juhan Bok, Dongmin Kim, Sagar Dattatraya Nale, Yun Sung Jo, Changjoong Kim, Taehhwan Park, Jaejune Dong, Byoung Gon Moon. Design, synthesis, and evaluation of next-generation EGFR degraders to overcome osimertinib-resistance [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 4502.

Oncogenic RAS mutations, commonly observed in human tumors, affect approximately 30% of cancer cases and pose a significant challenge for effective cancer treatment. Current strategies to inhibit the KRAS G12D mutation have shown limited success, emphasizing the urgent need for new therapeutic approaches. In this study, we designed and synthesized several purine and pyrimidine analogs as inhibitors for the KRAS G12D mutation. Our synthesized compounds demonstrated potent anticancer activity against cell lines with the KRAS G12D mutation, effectively impeding their growth. They also exhibited low toxicity in normal cells, indicating their selective action against cancer cells harboring the KRAS G12D mutation. Notably, the lead compound, PU1-1 induced the programmed cell death of KRAS G12D-mutated cells and reduced the levels of active KRAS and its downstream signaling proteins. Moreover, PU1-1 significantly shrunk the tumor size in a pancreatic xenograft model induced by the KRAS G12D mutation, further validating its potential as a therapeutic agent. These findings highlight the potential of purine-based KRAS G12D inhibitors as candidates for targeted cancer therapy. However, further exploration and optimization of these compounds are essential to meet the unmet clinical needs of patients with KRAS-mutant cancers.

Neuroblastoma (NB) is the most common extracranial childhood cancer, caused by the improper differentiation of developing trunk neural crest cells (tNCC) in the sympathetic nervous system. The N 6 -methyladenosine (m 6 A) epitranscriptomic modification controls post-transcriptional gene expression but the mechanism by which the m 6 A methyltransferase complex METTL3/METTL14/WTAP is recruited to specific loci remains to be fully characterized. We explored whether the m 6 A epitranscriptome could fine-tune gene regulation in migrating/differentiating tNCC. We demonstrate that the m 6 A modification regulates the expression of HOX genes in tNCC, thereby contributing to their timely differentiation into sympathetic neurons. Furthermore, we show that posterior HOX genes are m 6 A modified in MYCN-amplified NB with reduced expression. In addition, we provide evidence that sustained overexpression of the MYCN oncogene in tNCC drives METTL3 recruitment to a specific subset of genes including posterior HOX genes creating an undifferentiated state. Moreover, METTL3 depletion/inhibition induces DNA damage and differentiation of MYCN overexpressing cells and increases vulnerability to chemotherapeutic drugs in MYCN-amplified patient-derived xenografts (PDX) cells, suggesting METTL3 inhibition could be a potential therapeutic approach for NB.

IntroductionAlzheimer’s disease (AD) is the most studied progressive eurodegenerative disorder, affecting 40–50 million of the global population. This progressive neurodegenerative disease is marked by gradual and irreversible declines in cognitive functions. The unavailability of therapeutic drug candidates restricting/reversing the progression of this dementia has severed the existing challenge. The development of acetylcholinesterase (AChE) inhibitors retains a great research focus for the discovery of an anti-Alzheimer drug.Materials and methodsThis study focused on finding AChE inhibitors by applying the machine learning (ML) predictive modeling approach, which is an integral part of the current drug discovery process. In this study, we have extensively utilized ML and other in silico approaches to search for an effective lead molecule against AChE.Result and discussionThe output of this study helped us to identify some promising AChE inhibitors. The selected compounds performed well at different levels of analysis and may provide a possible pathway for the future design of potent AChE inhibitors.

Insights into host-virus interactions during SARS-CoV-2 infection are needed to understand COVID-19 pathogenesis and may help to guide the design of novel antiviral therapeutics. N-6-Methyladenosine modification (m(6)A), one of the most abundant cellular RNA modifications, regulates key processes in RNA metabolism during stress response. Gene expression profiles observed postinfection with different SARS-CoV-2 variants show changes in the expression of genes related to RNA catabolism, including m(6)A readers and erasers. We found that infection with SARS-CoV-2 variants causes a loss of m(6)A in cellular RNAs, whereas m(6)A is detected abundantly in viral RNA. METTL3, the m(6)A methyltransferase, shows an unusual cytoplasmic localization postinfection. The B.1.351 variant has a less-pronounced effect on METTL3 localization and loss of m(6)A than did the B.1 and B.1.1.7 variants. We also observed a loss of m(6)A upon SARS-CoV-2 infection in air/liquid interface cultures of human airway epithelia, confirming that m(6)A loss is characteristic of SARS-CoV-2-infected cells. Further, transcripts with m(6)A modification are preferentially down-regulated postinfection. Inhibition of the export protein XPO1 results in the restoration of METTL3 localization, recovery of m(6)A on cellular RNA, and increased mRNA expression. Stress granule formation, which is compromised by SARS-CoV-2 infection, is restored by XPO1 inhibition and accompanied by a reduced viral infection in vitro. Together, our study elucidates how SARS-CoV-2 inhibits the stress response and perturbs cellular gene expression in an m(6)A-dependent manner.

Objectives: Diabetes mellitus (DM), cancer and cardiovascular diseases (CVD) are major contributors to human miseries and death. Additionally, previous findings stated that cancer is also known to be associ-ated with diabetes. Therefore, in the given study, facile synthesis of gold nanoparticles (AuNPs) using gly-cation reaction was achieved and further, their bioconjugation with herbal drug garcinol (G) was performed (G-AuNPs) in order to enhance the antiglycative, antidiabetic and anticancer efficacy of the garcinol. Methods: The antiglycation and antidiabetic potential of garcinol bioconjugated AuNPs (G-AuNPs) were assessed through different experiments. Further, anticancer potential against adenocarcinoma cells were also assessed. Results: The study confirmed 78.6 % bioconjugation of G over the surface of AuNPs. Furthermore, the antiglycation study depicted that G-AuNPs are found to be more effective inhibitor of the glycation reac-tion in contrast to pure G. The findings represented that G-AuNPs significantly blocks the formation of early glycation adducts and AGEs, also they mask the glycation prone free arginine and lysine residues from participating in the glycation process. Additionally, our a-amylase inhibition assay results demon-strated that pure G (IC50 -8.9 lM) has significantly higher a-amylase inhibition activity compared to standard inhibitor, acarbose (IC50 -0.118 mM). Also, the anticancer study described that G and G-AuNPs treatment resulted in death of the cells via apoptosis, which resulted in elevated permeability, condensed chromatin, deep blue fluorescent, and condensed nucleus of A549 cells. Our findings also revealed that the potential of mitochondrial membrane (DWm) of A549 cells were also disrupted at much lower concentration of G-AuNPs (13.3 lM) as compared to pure G (28.7 lM). Conclusion: The key findings of the investigation suggested that G-AuNPs acted as a potent antiglycation, antidiabetic, and anticancer agent. (c) 2023 The Authors. Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Abstract Pathogenic coronavirus, including COVID-19, threatens human health, and there are strong demands for efficient therapeutics. Cordyceps militaris is a medicinal mushroom that has long been used for immune enhancement, anticancer, and antiviral effects. Therefore, the inhibitory potentials of constituents of C. militaris against COVID-19 were analyzed using various virtual screening analyses. Among ten constituents of C. militaris, cordycepin, the major component, and 3´-deoxyuridine and 2´-O-methyl-adenosine showed strong binding affinity to Mpro, a potential target for COVID-19 therapeutics. Considering the structure activity relationship, nucleosides having deoxyribose and methoxyribose moiety are important for the affinity to Mpro. Cordycepin is also bound to Mpro mutants, and the binding mechanisms between cordycepin and Mpro were further demonstrated by MD simulation, RMSA, and MM/PBSA analysis. Principal component analysis also confirmed the conformational change of Mpro by cordycepin, which inhibits the function of Mpro. In vitro, the efficacy of cordycepin was measured using Vero cells infected with SARS-CoV-2, which showed excellent inhibition with an IC50 value of 29 μM. Conclusively, the constituents of C. militaris are expected to inhibit SARS-CoV-2 replication through binding to Mpro. Therefore, C. militaris can be an important therapeutic for coronavirus through the synergistic effect of its constituents.

Myostatin (MSTN) is a well-reported negative regulator of muscle growth and a member of the transforming growth factor (TGF) family. MSTN has important functions in skeletal muscle (SM), and its crucial involvement in several disorders has made it an important therapeutic target. Several strategies based on the use of natural compounds to inhibitory peptides are being used to inhibit the activity of MSTN. This review delivers an overview of the current state of knowledge about SM and myogenesis with particular emphasis on the structural characteristics and regulatory functions of MSTN during myogenesis and its involvements in various muscle related disorders. In addition, we review the diverse approaches used to inhibit the activity of MSTN, especially in silico approaches to the screening of natural compounds and the design of novel short peptides derived from proteins that typically interact with MSTN.

Cyclin-dependent kinases (CDKs) play significant roles in numerous physiological, and are considered an attractive drug target for cancer, neurodegenerative, and inflammatory diseases. In the present study, we have aimed to investigate the binding affinity and inhibitory potential of selonsertib toward CDK6. Using the drug repurposing approach, we performed molecular docking of selonsertib with CDK6 and observed a significant binding affinity. To ascertain, we further performed essential dynamics analysis and free energy calculation, which suggested the formation of a stable selonsertib-CDK6 complex. The in-silico findings were further experimentally validated. The recombinant CDK6 was expressed, purified, and treated with selonsertib. The binding affinity of selonsertib to CDK6 was estimated by fluorescence binding studies and enzyme inhibition assay. The results indicated an appreciable binding of selonsertib against CDK6, which subsequently inhibits its activity with a commendable IC50 value (9.8 μM). We concluded that targeting CDK6 by selonsertib can be an efficient therapeutic approach to cancer and other CDK6-related diseases. These observations provide a promising opportunity to utilize selonsertib to address CDK6-related human pathologies.

Introduction: Selonsertib, the most recently developed selective inhibitor of apoptosis signal-regulating kinase 1. We elucidated the binding characteristics, mechanism of interaction, and dynamic behaviors of selonsertib with human serum albumin (HSA), a major circulatory transport protein. Methods: Different biophysical approaches (fluorescence quenching and isothermal titration calorimetry (ITC) were combined with various in silico techniques to examine the binding of selonsertib to HSA. Molecular docking results, analysis of molecular dynamics trajectories, and essential dynamics investigations indicated the stable binding of selonsertib to HSA. Further in vitro studies were performed to validate the observed interaction. Results: ITC results confirmed the robust binding and high affinity of selonsertib and HSA. Likewise, the fluorescence quenching results highlighted the binding affinity of selonsertib and HSA. Collectively, our findings offer deeper insight into the binding mechanism of selonsertib and HSA, emphasizing the selonsertib-mediated structural changes within HSA, along with a comprehensive rationale for the biological transport and accumulation of selonsertib in the blood plasma. Conclusion: Therefore, considering the bioavailability and effectiveness of selonsertib, assessing the interactions of this inhibitor with carrier proteins is crucial to elucidate its biological processes at the molecular level. This evidence carries the considerable scientific potential for future drug design.

Glioblastoma is the most aggressive form of brain tumor, accounting for the highest mortality and morbidity rates. Current treatment for patients with glioblastoma includes maximal safe tumor resection followed by ra-diation therapy with concomitant temozolomide (TMZ) chemotherapy. The addition of TMZ to the conformal radiation therapy has improved the median survival time only from 12 months to 16 months in patients with glioblastoma. Despite these aggressive treatment strategies, patients' prognosis remains poor. This therapeutic failure is primarily attributed to the blood-brain barrier (BBB) that restricts the transport of TMZ from reaching the tumor site. In recent years, nanomedicine has gained considerable attention among researchers and shown promising developments in clinical applications, including the diagnosis, prognosis, and treatment of glioblas-toma tumors. This review sheds light on the morphological and physiological complexity of the BBB. It also explains the development of nanomedicine strategies to enhance the permeability of drug molecules across the BBB.