Aslan, Osman Nuri; Kalın, Şeyda Nur; Kalay, Erbay; Türkeş, Cüneyt; Demir, Yeliz

In this study, a series of novel hybrid molecules ( TR1–14 ) incorporating a thiazole-hydrazine pharmacophore with a bicyclic isoindole-1,3-dione core were designed and synthesized. All compounds were unambiguously characterized by 1H/ 13C NMR, FTIR, and high-resolution mass spectrometry, and their purities (˃95%) were confirmed by HPLC. The synthesized derivatives exhibited potent α-glucosidase inhibition, with K I values ranging from 0.452 to 1.788 μM. Notably, the most active compound, TR6 ( K I: 0.452 ± 0.093 μM), demonstrated approximately 15-fold greater potency compared to the reference drug acarbose ( K I: 6.748 ± 0.167 μM), highlighting the substantial enhancement achieved through the designed scaffold modification. Structure–activity relationship analysis demonstrated that halogen substitution, particularly with chlorine and bromine, significantly enhances inhibitory potency, presumably due to strengthened hydrophobic and electronic interactions within the enzyme active site. Molecular docking and molecular dynamics simulations supported the experimental findings by revealing a conserved binding mode involving key catalytic residues (Arg600, Asp518, and Met519), as well as stabilizing water-mediated interactions, thereby providing a mechanistic rationale for the observed inhibition. In silico ADME predictions indicated favorable drug-like properties for all compounds, with full compliance with Lipinski's and Jorgensen's rules and substantially improved permeability profiles compared to acarbose. Cytotoxicity analyses demonstrated that TR3 , TR6 , TR10 , and TR11 exhibited low cytotoxic effects at concentrations above 100 μM in non-cancerous human lung epithelial BEAS-2B cells. Especially, TR5 and TR9 did not cause significant cytotoxicity even at a concentration of 200 μM in BEAS-2B cells. Overall, this integrated experimental-computational study identifies thiazole-hydrazine-isoindole hybrids, particularly TR5 and TR6 , as promising lead candidates for further optimization toward the development of next-generation α-glucosidase inhibitors with improved efficacy and drug-likeness.

El Hassani, Issam Ameziane; Altay, Ahmet; Yeniçeri, Esma; Boukharsa, Youness; Assila, Hamza; Alzahrani, Abdullah Yahya Abdullah; Moussa, Sana Ben; Ansar, M'’hammed; Karrouchi, Khalid

In this research, a series of novel pyrazole-benzofuran hydrazone hybrids (3a-h) were designed, synthesis and evaluated for their anticancer potential. All hybrid pyrazole-benzofuran derivatives were purified and characterized by using 1H NMR, 13C NMR, and ESI-HRMS analyses. Their cytotoxic potentials were evaluated against human lung carcinoma (A-549) and colorectal adenocarcinoma (HT-29) cell lines, and mouse fibroblast (3T3-L1) cells using XTT assay. The findings demonstrated that all compounds exhibited a dose-dependent anti-proliferative effect against the cancer cell lines under investigation. Compound 3d was particularly distinguished by its noteworthy IC50 value of 0.28 μM and a selectivity index of 103.85 on A549 cells. Advanced mechanistic anticancer investigations employing flow cytometry revealed that compound 3d triggered apoptosis in A549 cells by inducing mitochondrial membrane disruption and activating multiple caspases. Furthermore, both flow cytometry and Western blot analysis showed that the 3d molecule significantly suppressed the PI3K/AKT/mTOR signaling pathway, which is vital for cellular proliferation. In summary, the findings suggest that compound 3d has the potential to be evaluated as a therapeutic agent for the treatment of lung cancer.

Barlak, Neslisah; Sanli, Fatma; Celik, Suleyman; İskender, Burhan; Anil, Derya Aktas; Gambacorta, Nicola; Burmaoglu, Serdar; Nicolotti, Orazio; Algul, Oztekin; Karatas, Omer Faruk

Cancer continues to be a major worldwide health concern, with angiogenesis playing a pivotal role in its progression and metastasis. The capacity of tumors to induce angiogenesis is crucial for their proliferation and metastasis, rendering it a significant target for treatment strategies. This study involved the synthesis of sulfonamide-chalcone hybrid compounds and the evaluation of their anti-angiogenic activity by the assessment of their effects on vascular endothelial growth factor receptors (VEGFR1/VEGFR2) in human umbilical vein endothelial cells (HUVECs). Among the synthesized compounds, 28 and 31 exhibited the most promising inhibitory effects similar to sorafenib as positive control, with IC50 values superior to 10 μM. They dramatically diminished endothelial cell proliferation and tube formation without exhibiting significant cytotoxicity against healthy human cells. Molecular docking simulations validated their binding affinity to VEGFR1/VEGFR2 receptors, elucidating their molecular mechanisms. Moreover, compounds 28 and 31 exhibited significant anti-angiogenic activity in both 2D and 3D angiogenesis assays, indicating their potential as novel anti-cancer agents. These findings underscore the therapeutic potential of these compounds in blocking angiogenesis and necessitate additional in vivo research to confirm their clinical relevance.

Zognjani, Blinera; Nixha, Arleta Rifati; Duran, Hatice Esra; Arslan, Mustafa; Yıldıztekin, Gizem; Ece, Abdulilah; Türkeş, Cüneyt

Aldose reductase (ALR2; AKR1B1), a NADPH-dependent cytosolic oxidoreductase, plays a central role in the polyol pathway and is implicated in hyperglycemia-induced tissue injury. Beyond its metabolic function, elevated ALR2 expression has been reported in several malignancies, including hepatocellular and pulmonary carcinomas, highlighting its potential as a therapeutic target in metabolic-oncologic interface. In this study, a novel set of eleven N-substituted phthalimide–carboxylic acid derivatives (5a–5k) was synthesized and evaluated for ALR2 inhibition, pharmacokinetic characteristics, and cancer-selective safety. Among the series, compound 5f demonstrated the highest inhibitory potency (KI = 7.34 nM), outperforming epalrestat (KI = 232.1 nM). Glide docking positioned 5f within the ALR2 active site (GlideScore: −6.71 kcal/mol), stabilized via key contacts with Tyr48, His110, and Cys298, along with π–π stacking at Trp219. MM-GBSA analysis corroborated strong binding affinity (ΔG = −64.86 kcal/mol). DFT-derived quantum descriptors, logP, TPSA, and solvation energies supported its favorable interaction profile. ADME/Tox predictions indicated high GI absorption, no P-gp or CYP liabilities, and acceptable bioavailability. In vitro cytotoxicity assays showed negligible activity of 5f against A549 and Hep3B cancer cell lines (IC₅₀ > 160 μM) and no toxicity toward L929 fibroblasts, reflecting safety for long-term use. Transcriptomic data from CCLE and DepMap confirmed AKR1B1 overexpression in these cancer models. Network analysis linked ALR2 to redox imbalance and inflammation, suggesting its broader role in tumorigenesis.

Cihan, Cansu; Faydali, Nagihan; Erol, Meryem; Arpaci, Özlem Temiz; Muhammed, Muhammed Tilahun; Dilek, Esra

In this study, a new series of 2-substituted-5-methylsulfonylbenzoxazole derivatives (C1-C16) were successfully synthesized. The structures of the compounds were confirmed by FT-IR, 1H NMR, 13C NMR, and mass spectroscopy, and the results for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzyme inhibitory activity were reported as IC₅₀ and Ki values. The compounds' Ki values against AChE ranged from 7.55 ± 2.88 to 87.39 ± 11.67 nM, and their Ki values against BChE ranged from 3.23 ± 0.64 to 107.86 ± 13.38 nM. Accordingly, all compounds except C1, C2, C11, and C12 were found to be more effective than donepezil against AChE, and all compounds except C1, C5, and C12 were found to be more effective than donepezil against BChE. Molecular docking studies were performed for all compounds against the AChE and BChE enzymes. Conventional hydrogen bonds and π interactions (π-π stacked, π-π T-shaped, π-alkyl, π-sigma, π-cation) were observed in the interactions of the compounds with the target enzymes. Molecular dynamics (MD) simulation of complexes formed by compounds with the highest inhibition activity on the two enzymes affirmed formation of stable complexes. Additionally, the molecular electrostatic potential (MEP) map of all compounds and the extent to which the HOMO-LUMO energy gaps affect the reactivity, stability, and inhibitor profiles of these compounds were investigated using density functional theory (DFT) studies, a computational analysis method. The study proves that 2-substituted-5-methylsulfonylbenzoxazole derivatives exhibit potent and selective cholinesterase inhibition.

Gundogdu, Saliha; Duran, Hatice Esra; Arslan, Mustafa; Çetinkaya, Büşra Demir; Türkeş, Cüneyt

Metabolic reprogramming in cancer cells creates actionable vulnerabilities for precision oncology. Aldose reductase (ALR2, AKR1B1; EC 1.1.1.21), a cytosolic NADPH-dependent oxidoreductase, has emerged as a multifunctional player in tumor progression, drug resistance, and oxidative stress modulation, yet remains underexploited as a cancer target. In this study, we report the rational design, synthesis, and biological evaluation of a focused series of fluorenyl-phthalimide carboxylic-acid hybrids (5a-5l) as dual-acting agents with selective anticancer potential and potent ALR2 inhibition. Among them, compound 5a exhibited the strongest ALR2 inhibition with a KI of 8.71 ± 0.81 nM, far surpassing the clinical comparator epalrestat (KI = 232.10 ± 14.42 nM). In MCF-7 and A549 carcinoma cells, 5a demonstrated moderate antiproliferative activity (IC50 = 52.04 μM and 192.91 μM, respectively) while sparing normal bronchial epithelial cells, indicating tumor selectivity. Molecular docking and MM-GBSA calculations (ΔGbind = −44.02 kcal/mol) revealed a consistent bidentate hydrogen bonding network supporting high-affinity binding involving Tyr48, His110, and Trp111, further stabilized by crystallographic water molecules. To explore structure-activity relationships, a Random Forest regression model was developed using 24 descriptors derived from QikProp. The model achieved strong predictive performance (Q2CV = 0.71), and SHAP analysis identified water/gas partition coefficient, skin permeability, and human oral absorption as the most influential features shaping inhibitory potency. In silico ADME/Tox profiling supported the drug-like nature of compound 5a, with no major CYP or hERG liabilities predicted. Collectively, these findings position 5a as a first-in-class ALR2-targeted anticancer lead, introducing a chemically novel and mechanistically validated chemotype for further development within the framework of metabolically targeted precision therapies.

Güleç, Özcan; Duran, Hatice Esra; Arslan, Mustafa; Yıldıztekin, Gizem; Ece, Abdulilah; Türkeş, Cüneyt

Aldose reductase (ALR2) has emerged as a dual-function therapeutic target, critically involved in diabetic complications and cancer-related redox adaptation. In this study, a novel series of 15 chalcone-inspired heteroaryl–dihydronaphthalenone hybrids bearing indole, carbazole, or phenothiazine motifs (4a–4f, 8a–8e, 12a–12d) were rationally designed, synthesized, and systematically evaluated for their ALR2 inhibitory and anticancer properties. Enzymatic inhibition assays revealed low-nanomolar to low-nanomolar KI values, with compound 8b (KI = 3.59 nM, pKI = 8.44) emerging as the most potent inhibitor, outperforming the reference drug Epalrestat. SAR analysis highlighted the critical role of flexible alkyl side chains and polycyclic aromatic scaffolds in optimizing hydrophobic anchoring. Complementary in silico studies, including MM-GBSA binding energy calculations (ΔGbind = −56.37 kcal/mol), quantum descriptors (HOMO–LUMO gap, solvation energy), and ADME/Tox profiling (SwissADME, QikProp, ADMETlab), provided further mechanistic insight into selectivity and drug-likeness. Lead compound 8b also demonstrated selective antiproliferative activity toward A549 lung carcinoma cells (IC₅₀ = 197.6 μM), with no observed cytotoxicity in Hep3B hepatoma or L929 fibroblast lines. Molecular dynamics simulations confirmed binding stability, while target prediction analyses suggested low off-target risk. This multi-parametric evaluation underscores the translational potential of scaffold-tuned ALR2 inhibitors as oxidative stress–modulating agents with cancer-selective properties.

Güleç, Özcan; Türkeş, Cüneyt; Arslan, Mustafa; Demir, Yeliz; Dincer, Busra; Ece, Abdulilah; İrfan Küfrevioğlu, Ömer; Beydemir, Şükrü

Despite significant developments in therapeutic strategies, Diabetes Mellitus remains an increasing concern, leading to various complications, e.g., cataracts, neuropathy, retinopathy, nephropathy, and several cardiovascular diseases. The polyol pathway, which involves Aldose reductase (AR) as a critical enzyme, has been focused on by many researchers as a target for intervention. On the other hand, spiroindoline-based compounds possess remarkable biological properties. This guided us to synthesize novel spiroindoline oxadiazolyl-based acetate derivatives and investigate their biological activities. The synthesized molecules' structures were confirmed herein, using IR, NMR (1H and 13C), and Mass spectroscopy. All compounds were potent inhibitors with KI constants spanning from 0.186 ± 0.020 μM to 0.662 ± 0.042 μM versus AR and appeared as better inhibitors than the clinically used drug, Epalrestat (EPR, KI: 0.841 ± 0.051 μM). Besides its remarkable inhibitory profile compared to EPR, compound 6k (KI: 0.186 ± 0.020 μM) was also determined to have an unusual pharmacokinetic profile. The results showed that 6k had less cytotoxic effect on normal mouse fibroblast (L929) cells (IC50 of 569.58 ± 0.80 μM) and reduced the viability of human breast adenocarcinoma (MCF-7) cells (IC50 of 110.87 ± 0.42 μM) more than the reference drug Doxorubicin (IC50s of 98.26 ± 0.45 μM and 158.49 ± 2.73 μM, respectively), thus exhibiting more potent anticancer activity. Moreover, molecular dynamic simulations for 200 ns were conducted to predict the docked complex's stability and reveal significant amino acid residues that 6k interacts with throughout the simulation.

Rouzi, Khouloud; Altay, Ahmet; Bouatia, Mustapha; Yeniçeri, Esma; Islam, Mohammad Shahidul; Oulmidi, Afaf; El Karbane, Miloud; Karrouchi, Khalid

In this study, seven isoniazid-hydrazone derivatives (3a-g) were synthesized and their structures elucidated by chromatographic techniques, and then the antiproliferative effects of these compounds on various cancer cells were tested. The advanced anticancer mechanism of the most potent compound was then investigated. Antiproliferative activities of the synthesized compounds were evaluated on human breast cancer MCF-7, lung cancer A-549, colon cancer HT-29, and non-cancerous mouse fibroblast 3T3-L1 cell lines by XTT assay. Flow cytometry analysis were carried out to determine cell cycle distribution, apoptosis, mitochondrial membrane potential, multi-caspase activity, and expression of PI3K/AKT signaling pathway. The XTT results showed that all the title molecules displayed cytotoxic activity at varying strengths in different dose ranges, and among them, the strongest cytotoxic effect and high selectivity were exerted by 3d against MCF-7 cells with the IC50 value of 11.35 µM and selectivity index of 8.65. Flow cytometry results revealed that compound 3d induced apoptosis through mitochondrial membrane disruption and multi-caspase activation in MCF-7 cells. It also inhibited the cell proliferation via inhibition of expression of PI3K/AKT and arrested the cell cycle at G0/G1 phase. In conclusion, all these data disclosed that among the synthesized compounds, 3d is notable for in vivo anticancer studies.

Askin, Sercan; Tahtaci, Hakan; Türkeş, Cüneyt; Demir, Yeliz; Ece, Abdulilah; Akalın Çiftçi, Gülşen; Beydemir, Şükrü

Imidazole and thiadiazole derivatives display an extensive application in pharmaceutical chemistry, and they have been investigated as bioactive molecules for medicinal chemistry purposes. Classical carbonic anhydrase (CA) inhibitors are based on sulfonamide groups, but inhibiting all CA isoforms nonspecifically, thereby causing undesired side effects, is the main drawback of these types of inhibitors. Here we reported an investigation of novel 2,6-disubstituted imidazo[2,1-b][1,3,4]thiadiazole derivatives (9a-k, 10a, and 11a) and 2,5,6-trisubstituted imidazo[2,1-b][1,3,4]thiadiazole derivatives (12a-20a) that do not possess the zinc-binding sulfonamide group for the inhibition of human carbonic anhydrase (hCA, EC 4.2.1.1) I and II isoforms and also of acetylcholinesterase (AChE, EC 3.1.1.7). Imidazo[2,1-b][1,3,4]thiadiazoles demonstrated low nanomolar inhibitory activity against hCA I, hCA II, and AChE (KIs are in the range of 23.44–105.50 nM, 10.32–104.70 nM, and 20.52–54.06 nM, respectively). Besides, compound 9b inhibit hCA I up to 18-fold compared to acetazolamide, while compound 10a has a 5-fold selectivity towards hCA II. The synthesized compounds were also evaluated for their cytotoxic effects on the L929 mouse fibroblast cell line. Molecular docking simulations were performed to elucidate these inhibitors’ potential binding modes against hCA I and II isoforms and AChE. The novel compounds reported here can represent interesting lead compounds, and the results presented here might provide further structural guidance to discover and design more potent hCA and AChE inhibitors.