Growth Inhibitory Activity Research Articles

Synthesis of Azahelicenes via Friedel‐Crafts‐Type Ring‐Closing Reaction of Axially Chiral Compounds

Aza[6]helicene and aza[7]helicene were synthesized by Fridel‐Crafts‐type cyclization of axially chiral compounds. From the axially chiral compounds prepared by Fischer indole synthesis, aza[6]helicene and aza[7]helicene were synthesized. Aza[7]helicene was also synthesized from an axially chiral pyrrole compound prepared by Piloty‐Robinson‐type pyrrole synthesis through simultaneous cyclization on both sides of the pyrrole ring. Aza[6]helicene isomerized readily at room temperature, while aza[7]helicene isomerization at room temperature was less likely to proceed. The cell growth inhibitory activity of each enantiomer of N‐Me‐aza[7]helicene in HeLa cells was examined. Differences in the strength of activity between the enantiomers were observed.

Synthesis and Antibacterial Evaluation of Novel Small-Molecule Antibacterials of a Reduced Acridine Structure in S. aureus Strains Including MRSA.

The increasing antibacterial drug resistance remains a threat to global health with increasing mortality and morbidity. There is an urgent need to find novel antibacterials and develop alternative strategies to combat the increasing antibacterial drug resistance. <P> </P> Objective: We aimed to synthesize novel small-molecule antibacterials to evaluate the structuredependent antibacterial compound activities against S. aureus and MRSA. <P> </P> Method: Compounds were synthesized by primary N-alkylation to form alkyl acridinium salts that were further functionalized with substituted phenyl residues and finally purified by column chromatography. The antibacterial growth inhibition activity was determined as MIC value. <P> </P> Results: The substituent effects on the determined antibacterial growth inhibitory properties have been discussed. <P> </P> Conclusion: The best activities have been found for compounds with methoxy functions, exceeding the activities of reported novel antibacterial peptides. The compounds have also shown antibacterial drug-enhancing effects, which have been manifested as a reduction in the MIC values of the used antibiotics.

Rutinosides-derived from Sarocladium strictum 6-O-α-rhamnosyl-β-glucosidase show enhanced anti-tumoral activity in pancreatic cancer cells

BackgroundLow targeting efficacy and high toxicity continue to be challenges in Oncology. A promising strategy is the glycosylation of chemotherapeutic agents to improve their pharmacodynamics and anti-tumoral activity. Herein, we provide evidence of a novel approach using diglycosidases from fungi of the Hypocreales order to obtain novel rutinose-conjugates therapeutic agents with enhanced anti-tumoral capacity.ResultsScreening for diglycosidase activity in twenty-eight strains of the genetically related genera Acremonium and Sarocladium identified 6-O-α-rhamnosyl-β-glucosidase (αRβG) of Sarocladium strictum DMic 093557 as candidate enzyme for our studies. Biochemically characterization shows that αRβG has the ability to transglycosylate bulky OH-acceptors, including bioactive compounds. Interestingly, rutinoside-derivatives of phloroglucinol (PR) resorcinol (RR) and 4-methylumbelliferone (4MUR) displayed higher growth inhibitory activity on pancreatic cancer cells than the respective aglycones without significant affecting normal pancreatic epithelial cells. PR exhibited the highest efficacy with an IC50 of 0.89 mM, followed by RR with an IC50 of 1.67 mM, and 4MUR with an IC50 of 2.4 mM, whereas the respective aglycones displayed higher IC50 values: 4.69 mM for phloroglucinol, 5.90 mM for resorcinol, and 4.8 mM for 4-methylumbelliferone. Further, glycoconjugates significantly sensitized pancreatic cancer cells to the standard of care chemotherapy agent gemcitabine.ConclusionsαRβG from S. strictum transglycosylate-based approach to synthesize rutinosides represents a suitable option to enhance the anti-proliferative effect of bioactive compounds. This finding opens up new possibilities for developing more effective therapies for pancreatic cancer and other solid malignancies.

Abstract PO3-25-08: Artificial Intelligence-based screening of small molecule inhibitors of phosphoenolpyruvate carboxykinase 2 (PCK2), a novel cancer therapeutic target

Abstract Background: Metabolic rewiring is a hallmark of cancer that is often mediated by alterations in metabolic isozyme expression. Isozymes that are overexpressed in cancer and catalyze rate limiting steps in key metabolic processes are potential therapeutic targets. We previously identified phosphoenolpyruvate carboxykinase 2 (PCK2) as cancer dominant isoform, required for maintaining high metabolic activity and proliferation of lung, prostate, and breast cancer (BC) cells in vitro and in vivo. No PCK2 specific small molecule inhibitors exist. Methods: mRNA expression levels were assessed in the TCGA data. PCK2 MISSION shRNAs (Millipore) were used to knock-down expression of PCK2 in BC cells. Alterations in metabolic flux caused by PCK2 down-regulation were examined via 13C-labeling and mass spectrometry using a SCIEX 5500 QTRAP and SelexION instrument for mobility separation of metabolites. The AtomNetTM (Atomwise Inc.) deep convolutional neural network structure-based drug design software was used to identify candidate small molecule PCK2 inhibitors which were tested in PCK1 and PCK2 in vitro enzyme activity assays. The lead compound was assessed for drug-like properties by QickPropV4.0.001w, and its growth inhibitory activity was tested in MDA-MB-321 and BT-549 cells. Results: Analysis of mRNA levels in TNBC samples revealed a significant decrease in PCK1 expression compared to normal tissues, while PCK2 expression remained elevated or even increased. Quantification of mRNA levels in TNBC cell lines confirmed the higher expression of PCK2 compared to PCK1. To assess functional significance of PCK2 expression, we downregulated PCK2 using shRNA constructs in TNBC cell lines. Western blot analysis confirmed efficient knockdown of PCK2 protein without affecting PCK1 expression. Clonogenic assays demonstrated that PCK2-depleted BT-549 and MDA-MB-231 cells exhibited impaired proliferation, with reduced colony number and size compared to control cells. Metabolic analysis of PCK2-depleted TNBC cells demonstrated decreased levels of glycolytic and TCA intermediates and reduced flux through pyruvate carboxylase, indicating altered metabolic pathways upon PCK2 depletion. To identify potential inhibitors of PCK2, virtual screening was conducted using compound structures. We used a PCK2 protein structure homology model and designated the 3-mercaptopicolinic acid-binding site of the PCK1 structure as the site of interest to perform virtual drug screening of 7 million diverse compounds. The first screen yielded 86 structures to be tested in vitro, of which 5 demonstrated &amp;gt; 16% PCK2 inhibition. The top hit (IC50=2.4µM) was used to refine a second round of in silico screen that yielded 87 second-round candidates that were assessed for PCK2 inhibition in vitro. 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate emerged as the lead compound. QickProp predicted favorable absorption, distribution, metabolism, and excretion properties, but relatively low cell permeability (apparent Caco-2 and MDCK2 cell permeabilities 18 nm/sec and 8 nm/sec, respectively). The specificity of 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate was tested in PCK1 and PCK2 enzymatic assays and revealed IC50 of 500nM on PCK1 and an IC50 of 3 nM on PCK2. We next tested the cell growth inhibitory effect of 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate on parental MDA-MB-231 and BT-549 cells and their PCK2 knockdown clones. Cell growth and viability significantly decreased in the MDA-MD-231 and BT-549 parental cells (the IC50 for MDA-MB-231 was 173 μM) and in shRNA non-targeting control cells, but not in the PCK2 depleted clones, demonstrating PCK2-dependent inhibitory effect. Conclusion: We identified 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate as a high affinity selective PCK2 enzyme inhibitor. This compound also has significant growth inhibitory activity in BC cell lines in vitro and represents a novel therapeutic lead compound. Citation Format: Alejandro Rios Hoyo, Naing Lin Shan, Hao-Kuen Lin, Gerson Espinoza Campos, Mostafa Ahmed, Sheila Umlauf, Rebecca Cardone, Yulia V. Surovtseva, Vignesh Gunasekharan, Lajos pusztai. Artificial Intelligence-based screening of small molecule inhibitors of phosphoenolpyruvate carboxykinase 2 (PCK2), a novel cancer therapeutic target [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO3-25-08.

Design, synthesis, and biological evaluation of chalcone acetamide derivatives against triple negative breast cancer

Chalcones are chemical scaffolds found in natural products, particularly in plants, and are considered for structural diversity in medicinal chemistry for drug development. Herein, we designed and synthesised novel acetamide derivatives of chalcone, characterizing them using 1H NMR, 13C NMR, HRMS, and IR spectroscopic methods. These derivatives were then screened against human cancer cells for cytotoxicity using the SRB assay. Among the tested derivatives, 7g, with a pyrrolidine group, exhibited better cell growth inhibition activity against triple-negative breast cancer (TNBC) cells. Further assays, including SRB, colony formation, and fluorescent dye-based microscopic analysis, confirmed that 7g significantly inhibited MDA-MB-231 cell proliferation. Furthermore, 7g promoted apoptosis by upregulating cellular reactive oxygen species (ROS) levels and disrupting mitochondrial membrane potential (MMP). Elevated expression of pro-apoptotic proteins (Bax and caspase-3) and a higher Bax/Bcl-2 ratio with downregulation of anti-apoptotic (Bcl-2) protein levels were observed in TNBC cells. The above results suggest that 7g can promote cellular death through apoptotic mechanisms in TNBC cells.

Synthesis, growth inhibitory activity against tumor cells and structure-activity relationship of CGK733 and its analogs.

CGK733 was reported as a compound that inhibited ATM/ATR kinase activities and blocked their checkpoint signaling pathways with great selectivity. However, this paper was subsequently retracted, and the truth about the activity of CGK733 remains unclear. We synthesized various analogs of CGK733 with a modification of the carboxylic acid moiety and/or the aniline derivative moiety to accumulate knowledge of the structure-activity relationship of this compound. Growth inhibitory activity of CGK733 and novel 35 analogs against HeLa S3 cells was evaluated, and the structure-activity relationship revealed that analogs with 2-naphthyl or 4-fluorophenyl group instead of the benzhydryl group have activity comparable to CGK733 and that the 3-nitro group on the aniline moiety significantly affects the activity.

Repurposing of Antifungal Drug Flucytosine/Flucytosine Cocrystals for Anticancer Activity against Prostate Cancer Targeting Apoptosis and Inflammatory Signaling Pathways.

This study aimed to repurpose the antifungal drug flucytosine (FCN) for anticancer activity together with cocrystals of nutraceutical coformers sinapic acid (SNP) and syringic acid (SYA). The cocrystal screening experiments with SNP resulted in three cocrystal hydrate forms in which two are polymorphs, namely, FCN-SNP F-I and FCN-SNP F-II, and the third one with different stoichiometry in the asymmetric unit (1:2:1 ratio of FCN:SNP:H2O, FCN-SNP F-III). Cocrystallization with SYA resulted in two hydrated cocrystal polymorphs, namely, FCN-SYA F-I and FCN-SYA F-II. All the cocrystal polymorphs were obtained concomitantly during the slow evaporation method, and one of the polymorphs of each system was produced in bulk by the slurry method. The interaction energy and lattice energies of all cocrystal polymorphs were established using solid-state DFT calculations, and the outcomes correlated with the experimental results. Further, the in vitro cytotoxic activity of the cocrystals was determined against DU145 prostate cancer and the results showed that the FCN-based cocrystals (FCN-SNP F-III and FCN-SYA F-I) have excellent growth inhibitory activity at lower concentrations compared with parent FCN molecules. The prepared cocrystals induce apoptosis by generating oxidative stress and causing nuclear damage in prostate cancer cells. The Western blot analysis also depicted that the cocrystals downregulate the inflammatory markers such as NLRP3 and caspase-1 and upregulate the intrinsic apoptosis signaling pathway marker proteins, such as Bax, p53, and caspase-3. These findings suggest that the antifungal drug FCN can be repurposed for anticancer activity.

DB-1310, an ADC comprised of a novel anti-HER3 antibody conjugated to a DNA topoisomerase I inhibitor, is highly effective for the treatment of HER3-positive solid tumors

BackgroundHER3 (ErbB3), a member of the human epidermal growth factor receptor family, is frequently overexpressed in various cancers. Multiple HER3-targeting antibodies and antibody–drug conjugates (ADCs) were developed for the solid tumor treatment, however none of HER3-targeting agent has been approved for tumor therapy yet. We developed DB-1310, a HER3 ADC composed of a novel humanized anti-HER3 monoclonal antibody covalently linked to a proprietary DNA topoisomerase I inhibitor payload (P1021), and evaluate the efficacy and safety of DB-1310 in preclinical models.MethodsThe binding of DB-1310 to Her3 and other HER families were measured by ELISA and SPR. The competition of binding epitope for DB-1310 and patritumab was tested by FACS. The sensitivity of breast, lung, prostate and colon cancer cell lines to DB-1310 was evaluated by in vitro cell killing assay. In vivo growth inhibition study evaluated the sensitivity of DB-1310 to Her3 + breast, lung, colon and prostate cancer xenograft models. The safety profile was also measured in cynomolgus monkey.ResultsDB-1310 binds HER3 via a novel epitope with high affinity and internalization capacity. In vitro, DB-1310 exhibited cytotoxicity in numerous HER3 + breast, lung, prostate and colon cancer cell lines. In vivo studies in HER3 + HCC1569 breast cancer, NCI-H441 lung cancer and Colo205 colon cancer xenograft models showed DB-1310 to have dose-dependent tumoricidal activity. Tumor suppression was also observed in HER3 + non-small cell lung cancer (NSCLC) and prostate cancer patient-derived xenograft (PDX) models. Moreover, DB-1310 showed stronger tumor growth-inhibitory activity than patritumab deruxtecan (HER3-DXd), which is another HER3 ADC in clinical development at the same dose. The tumor-suppressive activity of DB-1310 synergized with that of EGFR tyrosine kinase inhibitor, osimertinib, and exerted efficacy also in osimertinib-resistant PDX model. The preclinical assessment of safety in cynomolgus monkeys further revealed DB-1310 to have a good safety profile with a highest non severely toxic dose (HNSTD) of 45 mg/kg.ConclusionsThese finding demonstrated that DB-1310 exerted potent antitumor activities against HER3 + tumors in in vitro and in vivo models, and showed acceptable safety profiles in nonclinical species. Therefore, DB-1310 may be effective for the clinical treatment of HER3 + solid tumors.

An In Vitro Study on the Antibacterial Activity of Statins Against the Most Common Bacterial Pathogens Causing Respiratory Tract Infections in a Tertiary Care Hospital.

Introduction The vast pleiotropic effect of statins has intrigued many researchers to select them as potential candidates against bacterial infections. The role of statins against bacterial pathogens remains debatable. This study was undertaken to evaluate and compare the antibacterial effect of commonly available statins against the most frequently isolatedbacterial pathogens causing respiratory tract infections in our tertiary care hospital using sputum as a sample. Materials and methods The study was conducted in the Microbiology Laboratory of our hospital. Drugs including atorvastatin, rosuvastatin, and simvastatin were purchased in pure form from Sigma Aldrich. Dimethylsulfoxide (DMSO) was used as a solvent for all three drugs. The positive controls used were gentamycin and amoxicillin for Gram-negative and Gram-positive bacteria, respectively. Data regarding all the culture and sensitivity results of sputum samples of patients admitted to the Respiratory Intensive Care Unit over the past 12 months were analyzed. The most common bacterial pathogensStaphylococcus aureus, Klebsiella pneumoniae,and Streptococcus pneumoniae isolated from sputum specimens were taken for our study. The antibacterial effect of statins was studied using two methods: the agar cup diffusion method and the broth dilution method. The zone of inhibition and minimum inhibitory concentration of the drugs were calculated and analyzed. Statistical analysis was performed using GraphPad Prism software version 10.2.0. A one-way ANOVAtest was used to determine if there was any statistical difference between the different statins and antibiotic groups. An unpaired t-test was used to determine the statistical difference between the statins. Results and discussion For the agar cup diffusion method, our results displayed a lack of antibacterial activity of all three statinsatorvastatin, rosuvastatin, and simvastatin against all three bacterial strainsStaphylococcus aureus, Klebsiella pneumoniae,and Streptococcus pneumoniae after overnight incubation by agar cup method at concentrations of3.125 μg/ml, 6.25 μg/ml, 12.5 μg/ml, 25 μg/ml and 50 μg/ml, respectively. The zone of inhibition observed was less than 4 mm (resistant) for all the serial dilutions of atorvastatin, rosuvastatin, and simvastatin. For the broth dilution method, the ANOVA test showed amoxicillin and gentamicin to have high statistically significant microbial growth inhibitory activity (p-value< 0.005) compared to atorvastatin and rosuvastatin. Statistically, though atorvastatin showed significant antimicrobial activity compared to normal saline and rosuvastatin,thiswas not considered clinically significant as the antimicrobial activity shown by atorvastatin was very negligible compared to the controls used and did not correspondto the serial dilutions of the drug. Conclusion Atorvastatin, rosuvastatin, and simvastatin lacked antibacterial activity against all three bacterial strains isolated from sputum specimens:Staphylococcus aureus, Klebsiella pneumoniae, and Streptococcus pneumoniae.Hence, the use of statins as an antimicrobial drug for respiratory tract infections has limited applications.

Synthesis and Cytotoxic Studies of Pyrrole and Pyrrolidine Derivatives in Human Tumor Cell Lines

AbstractHeterocyclic compounds such as pyrrole and pyrrolidine derivatives have a broad spectrum of biological activity, being widely used as pharmacophore to design novel bioactive compounds. In this work, sixteen pyrrole and pyrrolidine derivatives were synthesized and evaluated for their growth inhibitory activity on two human cancer cells lines: breast cancer MDA‐MB‐231 and chronic myelogenous leukemia (K562). Eight compounds showed activity against at least one tumor cell line (IC50=49.15–195.9 μM). The compounds were tested in non‐cancerous human lung fibroblasts WI‐26VA4 to evaluate their selectivity index. In addition, Hierarchical Cluster Analysis (HCA) studies were carried out in attempt to establish a structure‐activity relationship.

Combined computational and synthetic strategies for the development of potent pyrazolo-pyridine derivatives as anticancer agents

PI3K comprise a group of lipid kinases mainly made up of three classes and have a particular role in signal transduction. The aberration of class IA PI3K is most frequently shown in cancer. The computation techniques have been applied for the identification of potent compounds. This involved the generation of a library of novel compounds (2700) through R-group enumeration study. The screening process utilized a combination of pharmacophore-based virtual screening and various docking algorithms, leading to the discovery of ten novel compounds. The docking studies against PI3K kinase highlighted MSA6 and MSA7 as the most promising compounds, with docking scores of -9.03 and -9.0 kcal/mol, respectively. Results of docking studies of all derivatives confirmed satisfactory binding mode of the compounds within the active site of the target protein PI3K kinase. The screened pyrazolo-pyridine derivatives (MSA1-MSA10) were synthesized and evaluated as anti-cancer agents against NSCLC using A549 cell line. MSA6 showed the dose dependent enhancement in ROS production at concentration of 10 and 20 µM in A549 cells. MSA6 triggered caspase 3 cleavage in lung cancer cells in a dose-dependent manner. MSA6 treatment at 10 and 20 µM levels boosted NF-κβ expression in the cytoplasm while decreasing it in the nucleus, as demonstrated by flow cytometry with an anti-NF-κβ antibody. The modulation of PI3K activity have been observed at 10 µM and 20 µM concentration of MSA6. A549 cells treated with MSA6 with 20 µM showed significant reduction in wound healing in cancerous cell. A549 cells treated with 20 µM showed significantly reduction in number of colonies compared to control group. Compound MSA6 having a methyl group at the para position of phenyl ring at imine carbon and phenyl ring substitution at nitrogen of pyrazole ring exhibited remarkable growth inhibitory activity with IC50 value of 49.23 μM. Based on the above results, pyrazolo-pyridine derivatives can be used for the further development of novel PI3K kinase inhibitors as anticancer agents.

A promising perspective through potential antimicrobial, anti‐cancer, and antiviral activity against SARS‐CoV‐2 of silver(I) complexes: Synthesis, characterization, density functional theory calculations, and in‐silico molecular docking studies

This study tends to accomplish the sustainable development goal, which calls for all healthy lives. Antibiotic resistance, cancer, and severe acute respiratory syndrome coronavirus all showed high death rates in recent years. Our approach is to make better medications available to fight these feared health issues. In this context, three silver(I) complexes (1–3) based on the antibiotic gemifloxacin (HGMF) were synthesized and thoroughly examined using analytical, spectral, and thermal methods. The density functional theory suggested that complex 1 has a bent coordination environment while 2 and 3 have a distorted trigonal planar geometry. The optimal donor and acceptor centers were determined by the natural‐bond orbital analysis. Interaction of complexes with calf‐thymus DNA has been investigated and their binding constants were ordered as 3 &gt; 2 &gt; 1. The antimicrobial activity against 10 pathogenic microorganisms demonstrated that complex 3 exhibited superior activity compared with standard antibiotics. Testing against mammalian cell lines, liver carcinoma, breast carcinoma, and colon carcinoma revealed that complex 3 showed remarkable cell growth inhibition activity with IC50 values ranging from 3.0 to 9.3 μg/mL. Studies of Molecular docking against SARS‐CoV‐2 main protease Mpro (ID: 6 WTT) proved a powerful type of interaction at the active site. Complex 3 exhibits a potent inhibition effect with a total binding energy of −12.58 kcal/mol compared with the reference ligand inhibitor K36 suggesting its ability to block viral protease. This study presents promising bioactive candidates that have a practical impact on various medicinal fields.

Urease and human glioblastoma (U87) cells growth inhibitory iridoid glycoside acetates from Nyctanthes arbor-tristis Linn.

The present study was undertaken on the chemical constituents of ethanol extract of aerial parts of Nyctanthes arbor-tristis Linn., and their determination of growth inhibitory activity against glioblastoma multiforme cell line (U87) and urease inhibitory activity. Six constituents were isolated including two new arbortristoside F tetraacetate (1) and arbortristoside G heptaacetate (2) and four known arborside A tetraacetate (3), arborside C pentaacetate (4), 6,7-di-O-acetyl-6β-hydroxyloganin hexaacetate (5) and nyctanthoside heptaacetate (6) iridoid glycoside acetates. Their structures were elucidated using spectroscopic methods, including 1D and 2D NMR and mass analyses. Compounds 3 and 6 showed significant growth inhibition of U87 cell line (76.41 and 87.62% inhibition) respectively while 2, 4 and 5 showed moderate inhibition. 3 and 6 showed notable urease inhibition (IC50 = 17.2 ± 0.4 and 17.2 ± 0.7 µM) respectively, and IC50 of 2, 4 and 5 were 23.8 − 56.3 µM. Compound 1 was inactive.

Abstract LB126: Generation of a novel ROR1 x CD3 bispecific T-cell engager for better tumor killing and minimal cytokine release

Abstract Receptor tyrosine kinase-like orphan receptor 1 (ROR1) expression is a highly specific tumor associated antigen expressed in multiple hematological malignancies and solid tumors. EMB-07 is a novel monovalent asymmetrical tandem Fab (MAT-Fab) antibody composed of two different Fab fragments targeting ROR1 and CD3ε respectively, and a “knob-in-hole” Fc fragment maintaining FcRn binding and Ig-like pharmacokinetics in vivo. To avoid light chain miss-pairing, the anti-ROR1 light chain was directly connected to the N-terminus of the anti-CD3 heavy chain. The ROR1 binding arm has a high affinity of 0.52 nM and recognizes a membrane distal epitope, which is different from a reference BiTE molecule currently in clinical trial. In comparison with this BiTE molecule, EMB-07 showed similar tumor killing efficacy to ROR1 expressing tumor cell lines while it induced much lower levels of cytokines both in vitro and in vivo. Moreover, comparing with a clinical stage ROR1-vc-MMAE ADC, EMB-07 demonstrated ~500-fold increased tumor lysis potency in vitro. In PD-1 resistant tumor xenograft mice models, EMB-07 showed potent tumor growth inhibition and synergistic activity in combination with PD-1 blockade. In a preclinical GLP toxicological study, EMB-07 was well tolerated in cynomolgus monkeys with no evidence of lethality, life-threatening toxicity, or obvious EMB-07-related macroscopic and histopathological adverse findings up to 50 mg/kg. By using a highly specific IHC clone, we validated membrane-associated ROR1 expression in multiple tumor types, including triple negative breast cancer, ovarian cancer, endometrial cancer, lung adenocarcinoma, and pancreatic cancer, while the observation of its expression in normal tissues is very limited. This study supports the clinical development of EMB-07 for treating multiple solid tumors with ROR1 expression. A FIH study of EMB-07 in locally advanced/metastatic solid tumors or relapse/refractory lymphoma is currently ongoing (NCT05607498). Citation Format: Danqing Wu, Shiyong Gong, Xuan Wu, Gaowa Naren, Liqin Dong, Stephan Lensky, Chengbin Wu. Generation of a novel ROR1 x CD3 bispecific T-cell engager for better tumor killing and minimal cytokine release [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB126.

Abstract CT066: Phase 1 study of MDM2 antagonist ASTX295 in patients with solid tumors with wild-type TP53

Abstract Background: ASTX295 is a novel small molecule antagonist of human Murine Double Minute 2 (MDM2), which tightly regulates the level and activity of the p53 tumor suppressor. To minimize potential thrombocytopenia previously seen with antagonism of this target, ASTX295 was designed to have a short half-life (t1/2) with decreased bone marrow (BM) exposure. ASTX295 has proapoptotic and tumor growth inhibitory activity in preclinical cancer (CA) models. We report here the safety and preliminary efficacy of the phase 1 and dose expansion portions of Study ASTX295-01. Methods: ASTX295-01 is a first-in-human, open-label, multicenter, phase 1/2 study to assess safety, pharmacokinetics (PK), pharmacodynamics (PD), and preliminary clinical activity of ASTX295 (NCT03975387). Eligible patients (advanced solid tumors with Wild-Type [WT] TP53, ECOG PS 0-2, adequate organ and bone marrow function, etc.) had oral administration of ASTX295 starting at a daily dose of 15 mg. Dose escalation was by standard 3 + 3 with cohorts testing multiple regimens. Safety was assessed by CTCAE and disease was assessed by RECIST 1.1. Safety and efficacy and dosing recommendations were overseen by the Data Safety Review Committee. Results:83 subjects were enrolled in over 14 cohorts, evaluating daily (QD) and intermittent dosing regimens and the effect of food. The mean age of subjects on study was 60.1 year, 56.9 were male, with 2.94 (0-15) prior chemotherapy regimens. For PK exposure, the total weekly AUC for 400 mg QD and 660 twice a week (BIW) are similar. The median tmax is 3 hours and mean t1/2 is 4-6 hours. PD demonstrates p53 pathway modulation. Nausea, vomiting, diarrhea, and fatigue were dose limiting. RP2D regimens were 660 mg BIW and 400 mg QD and these regimens were expanded with a total of 25 additional subjects. The most common treatment related AEs for both regimens were nausea, diarrhea, and vomiting at 76.2%,76.2%, and 33.3% respectively, for daily and 71.4%, 47.6%, and 33.3%, for intermittent. Grade 3 GI AEs were reported in 14.3% of subjects in the daily cohort vs 9.5% in the intermittent; none had Grade ≥ 4. One subject had grade 2 thrombocytopenia. Based on decreased incidence and duration of GI AEs, 660 mg BIW was selected as the RP2D.Multiple objective responses were seen, including 1 subject with non-small cell lung CA and 3 subjects with liposarcoma (LPS), a tumor with frequent MDM2 amplification. For LPS the ORR is 7.9% (de-differentiated 7.7%, well differentiated 8.0%), with a PFS of 7.95 and 9.66 months respectively, along with a 16-week disease control rate of 69.2% and 72.0%. Five subjects with GBM were enrolled, 4 with MDM2 amplification, 3 of whom showed evidence of tumor regression, with one subject remaining on study for &amp;gt; 34 cycles. Conclusion:ASTX295 was well tolerated at doses producing p53 pathway modulation with manageable toxicities, notably avoiding significant thrombocytopenia. Preliminary single agent efficacy was observed in heavily pretreated subjects across multiple solid tumor types with WT TP53. Citation Format: Ecaterina E. Dumbrava, Fabio M. Iwamoto, Mark Agulnik, Mohammed Milhem, Anthony W. Tolcher, Rashmi Chugh, Michael J. Demeure, Alain Mita, Kurt Demel, Mark Diamond, Beloo Mirakhur, Laksmi Wilson, Aram Oganesian, Danna Chan, Harold N. Keer, Jason Taylor, Martin J. Sims, Andrea Biondo, Simone Jueliger, Alexander Spira. Phase 1 study of MDM2 antagonist ASTX295 in patients with solid tumors with wild-type TP53 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT066.

Evaluation of volatile components from the tuber, fibrous roots, bud, stem and leaf tissues of Bletilla striata for its anti-colon cancer activity.

Bletilla striata (Thunb.) Rchb.f., a medicinal plant in the Orchidaceae family, is mainly found in East Asia and has extensive pharmacological activities. Plant's volatile components are important active ingredients with a wide range of physiological activities, and B. striata has a special odor and unique volatile components. Yet it has received little attention, hindering a full understanding of its phytochemical components. Employing the ultrasonic-assisted extraction method, the volatile components of B. striata's fibrous root, bud, aerial part and tuber were extracted, resulting in yields of 0.06%, 0.64%, 3.38% and 4.47%, respectively. A total of 78 compounds were identified from their chemical profiles using gas chromatography-mass spectrometry (GC-MS), including 45 components with the main compounds of linoleic acid (content accounting for 31.23%), n-hexadecanoic acid (13.53%), and octadecanoic acid (9.5%) from the tuber, 34 components with the main compounds of eicosane, 2-methyl- (28.42%), linoelaidic acid (10.43%), linoleic acid (4.53%), and n-hexadecanoic acid (6.91%) from the fibrous root, 38 components with the main compounds of pentadeca-6,9-dien-1-ol (9.29%), n-hexadecanoic acid (11%), eicosane,2-methyl- (23.43%), and linoleic acid (23.53%) from the bud, and 27 components with the main compounds of linoelaidic acid (5.97%), n-hexadecanoic acid (15.99%), and linolenic acid ethyl ester (18.9%) from the aerial part. Additionally, the growth inhibition activity against colon cancer HCT116 cells was evaluated using sulforhodamine B (SRB) assay and the thiazolyl blue tetrazolium bromide (MTT) assay, and the accumulation of reactive oxygen species (ROS) was determined using dichloro-dihydro-fluorescein diacetate (DCFH-DA) staining and fluorescence intensity analysis. The volatile extracts exhibited significant growth inhibitory efficacy against HCT116 cells, with half-maximal inhibitory concentration (IC50) values of 3.65, 2.32, 2.42 and 3.89mg/mL in the SRB assay, and 3.55, 2.58, 3.12 and 4.80mg/mL in the MTT assay for the root, bud, aerial part, and tuber, respectively. Notably, treatment with the aerial part extract caused morphological changes in the cells and significantly raised the intracellular ROS level. In summary, the chemical profiles of the volatile components of B. striata were revealed for the first time, demonstrating a certain tissue specificity. Additionally, it demonstrated for the first time that these volatile extracts possess potent anti-colon cancer activity, highlighting the importance of these volatile components in B. striata's medicinal properties.

Design, Synthesis and Anticancer Activity of Substituted 1, 3-Thiazolidin-4-One Derivatives

Background: Cancer is the global cause of death worldwide. Anticancer drug development is the need in today’s scenario. Thiazolidine is the nucleus that shows several pharmacological activities like anticancer, anti-inflammatory, antioxidant, antibacterial, antifungal, antidiabetic, antihyperlipidemic, and antiarthritic activity. In the present work molecular docking Glide module (Schrodinger Inc., USA) has been used for ligand docking against the Polo-like kinase-1. The series of substituted 3-Benzothiazol-2-yl-2-phenyl)-thiazolidin-4-ones were synthesized by the microwave-assisted synthesis system (CEM, USA) and characterized by melting point, FT-IR, 1H NMR, 13C NMR, and HR-MS analysis. Results: Molecular docking studies shows good docking score as well as interactions. Among the synthesized compounds, BG2 had the highest docking score of -8.381, followed by BG8 (-8.19) and BG1 (-8.156). All the newly synthesized compounds were examined for their in vitro anticancer activity against breast cancer cell line MCF-7 by Sulforhodamine B (SRB) assay. Conclusion: BG1, BG2, BG3, BG4, BG5, BG8, BG9, BG10, BG11, BG12, and BG13 (GI50: &lt;80 µg/ml) exhibited significant cell growth inhibitory activity. These results indicate that compounds showing in-vitro activity by molecular docking studies and SRB assay could be lead compounds for further development of anticancer agents and suitable candidates for in-vivo anticancer activity.

Molecular Docking, Toxicity Study and Antimicrobial Assessment of Novel Synthesized 1,3-(Disubstituted)-thiazol-2-amines

In current study, a novel analogous of substituted-2-aminothiazoles (3a-o) were synthesized through a multi-step synthetic process. Structural elucidation of these newly synthesized substituted-2-aminothiazoles were achieved using combination of analytical techniques, comprising proton nuclear magnetic resonance (PNMR), mass spectrometry and FTIR. An in vitro investigation was performed to measure the efficacy of antibacterial and antimycotic characteristics of these novel compounds (3a-o). Specifically, the growth-inhibiting action against the test fungal strains, including A. niger, M. purpureos and A. flavus was examined. Additionally, their inhibitory antibacterial activity against key bacterial strains, including P. aeruginosa, S. aureus and E. coli was ascertained employing the agar diffusion technique. The results of antibacterial screening disclosed that maximum number of the thiazole derivatives viz. 3a, 3d, 3e, 3i, 3k, 3l and 3n displayed minimum inhibitory concentration of 12.5 µg/mL for E. coli. While compounds 3k and 3n displayed minimum inhibitory concentration of 12.5 µg/mL for S. aureus. A minimum inhibitory concentration of 25 µg/mL was exhibited by compounds 3i, 3l and 3n against P. aeruginosa. None of the 2-aminothiazole derivative disclosed promising action against the fungal strains. Screening for in silico ADME and toxicity studies revealed that compounds are fairly compatible and were devoid of potential toxicity except compounds 3j and 3m. The docking studies on DNA gyrase (PDB ID; 1KZN) shows favourable binding interaction comparable to the pre-occupied ligand clorobiocin.

Variations in Essential Oil Biological Activities of Female Cones at Different Developmental Stages from Azorean Cryptomeria japonica (Thunb. ex L.f.) D. Don (Cupressaceae)

In the Azores Archipelago, Cryptomeria japonica is, currently, the most cultivated forestry tree for timber production, landscaping, and gardening, generating large amounts of foliage waste that is used for local essential oils (EOs) production. However, the existing literature on the biological potential of EOs from different C. japonica foliage parts, such as female cones (FC), remains limited. Thus, in the present study, EOs extracted by hydrodistillation from Azorean C. japonica immature and mature FC (IFC and MFC), as well as some major EO components, were screened for their: (i) antioxidant capacity, evaluated by DPPH free-radical-scavenging activity (FRSA) and β-carotene-linoleic acid bleaching activity (BCBA), (ii) antimicrobial activities, assessed by the disc diffusion method against eight bacteria and one fungus, and (iii) toxicity against Artemia salina. Among both FC EO samples, the IFC EO exhibited the best DPPH–FRSA, BCBA, and growth inhibitory activity against Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, and Penicillium italicum, as well as a slightly increased toxic potential, due to their differential compositions, as assessed by GC–MS analysis. Thus, the FC’s maturation process decreased their EOs’ bioactivities. In conclusion, this finding could help in determining the optimal developmental stage for enhancing the antioxidant and antimicrobial compounds content in FC EOs. In turn, this contributes to increasing the commercial potential of C. japonica’s EO industry.

Discovery of Novel Pyrazole Acyl Thiourea Skeleton Analogue as Potential Herbicide Candidates.

To discover novel transketolase (TKL, EC 2.2.1.1) inhibitors with potential herbicidal applications, a series of pyrazole acyl thiourea derivatives were designed based on a previously obtained pyrazolamide acyl lead compound, employing a scaffold hopping strategy. The compounds were synthesized, their structures were characterized, and they were evaluated for herbicidal activities. The results indicate that 7a exhibited exceptional herbicidal activity against Digitaria sanguinalis and Amaranthus retroflexus at a dosage of 90 g ai/ha, using the foliar spray method in a greenhouse. This performance is comparable to that of commercial products, such as nicosulfuron and mesotrione. Moreover, 7a showed moderate growth inhibitory activity against the young root and stem of A. retroflexus at 200 mg/L in the small cup method, similar to that of nicosulfuron and mesotrione. Subsequent mode-of-action verification experiments revealed that 7a and 7e inhibited Setaria viridis TKL (SvTKL) enzyme activity, with IC50 values of 0.740 and 0.474 mg/L, respectively. Furthermore, they exhibited inhibitory effects on the Brassica napus acetohydroxyacid synthase enzyme activity. Molecular docking predicted potential interactions between these (7a and 7e) and SvTKL. A greenhouse experiment demonstrated that 7a exhibited favorable crop safety at 150 g ai/ha. Therefore, 7a is a promising herbicidal candidate that is worthy of further development.