Identification thiazole-based analogs as potent antidiabetic agents and their molecular docking study

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Alharamlah, Hafiz Ur Rehman, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8601560/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 7 You are reading this latest preprint version Abstract In this research work fifteen thiazole-based scaffolds ( 1–15 ) were synthesized and evaluated for the α-amylase and α-glucosidase inhibitory potential. In this series all the synthesized scaffolds showed good to moderate α-amylase and α-glucosidase inhibition and their IC 50 value ranging between 3.10 ± 0.10 µM to 15.40 ± 0.50 µM for α-amylase and IC 50 value from 3.30 ± 0.10 µM to 16.40 ± 0.60 µM for α-glucosidase when compared with the standard acarbose having IC 50 value 10.30 ± 0.20 µM (for α-amylase) and 9.80.20 ± 0.20 µM (for α-glucosidase). Among the synthesized scaffolds, nine analogs ( 1 – 3 , 6 – 9 , 12 and 13 ) were found more active against both enzymes. The scaffolds 1 IC 50 value (3.10 ± 0.10 µM, 3.30 ± 0.10 µM), 2 IC 50 value (4.50 ± 0.20 µM, 4.70 ± 0.20 µM) and 3 IC 50 value (4.20 ± 0.10 µM, 5.10 ± 0.20 µM) were found most potent among this series against both α-amylase and α-glucosidase enzymes respectively. These scaffolds having fluorine at ortho , meta and para position of the phenyl ring respectively. Furthermore, the structures of all the synthesized analogs were confirmed by using 1 H-NMR, 13 C-NMR spectroscopy and HR-MS. To study the binding mode of interaction between active site of the targeted enzyme and most potent scaffold, molecular docking study were conducted. Biological sciences/Biochemistry Biological sciences/Chemical biology Physical sciences/Chemistry Biological sciences/Drug discovery Thiazole antidiabetic activity SAR molecular docking Figures Figure 1 Figure 2 Figure 3 Introduction In today's world, the development of diabetes mellitus (DM) has been identified as one of the greatest health issues worldwide. In 2011 information collected by WHO there are approximately 366 million patients affected from diabetes and by 2030 the affected people increases to 522 million worldwide 1 . Type 1 diabetes is a chronic condition in which the immune system attacks the insulin producing pancreatic beta cells. Thus to maintain the level of the pancreatic beta cells, insulin administered to cure type 1 diabetes 2 . While diabetes-II is caused when beta cells of the pancreas secrete insufficient amount of insulin 3 . The α-amylase is the digestive enzymes that catalyze the hydrolysis of carbohydrate that result in blood sugar. The alpha amylase is secreted by pancreas and salivary gland and is responsible for the hydrolysis of starch and oligosaccharide into sugar. Alpha-glucosidase enzyme, which is responsible for the breakdown of polysaccharides is present on the brush boarder of small intestine 4 . The inhibition of these enzymes plays important role in controlling blood glucose by decreasing carbohydrate metabolism 5 . Several anti-diabetic drugs are used in clinical practice nowadays such as acarbose, voglibose and miglitol that reduce the activity of α-glucosidase enzyme which then consequently interfere the carbohydrate metabolism and reduces the chance of postprandial hyper-glycaemia and hyper- insulinemia 6 , 7 . However, the use of these drugs has adverse side effect such as flatulence, abdominal pain, dizziness and diarrhea, because of microorganisms in the colon fermenting excessive carbohydrates. In rare instances, intestinal difficulties such as pneumatosis cystoids intestinalis, liver problems, particularly with acarbose, and allergic skin reactions are less frequent but more dangerous adverse effects 8 . Therefore, there is intense need to develop more potent and efficient drugs on priority basis having low side effects. Thiazole is a heterocyclic molecule having numerous pharmacological applications such as inflammatory 8 hypertension 9 and bacterial disease 10 . Amino-thiazoles are popular as estrogen receptors ligands 11 and an innovative class of adenosine receptor antagonists 12 . Thiazole moiety is present in numerous medications such as in Fanetizole an anti-inflammatory drug, Abafungin an antifungal agent, Famotidine a histamine H2 receptor antagonist, and Pamicogrel an anti-cancer agent 13 . The thiazole analogs were reported as antidiabetic and anti-Alzheimer agents 14 – 17 . The method of molecular hybridization is an important approach to research and development of new biologically active compounds, which involves the hybridization of two different active substances from the same molecule. Due to the wide range of pharmacological activities associated with thiazoles and thiourea moieties, a type of thiazole-thiourea derivative has been designed and studied for the inhibition of diabetes mellitus. The aim of this study is to identify the best thiazole-thiourea derivatives as inhibitors of type 2 and study interaction of enzymes (α-amylase and α-glucosidase) with active compounds via molecular docking study. Results and discussion Chemistry Fifteen ( 1 – 15 ) thiazoles bearing thiourea analogs were synthesized by reacting 2-(2-bromophenyl) thiazole-5-carbohydrazide (1 mmol) with various aryl isothiocyanates (1 mmol) in chloroform (10 mL) as a solvent and allowing the reaction mixture to proceed for 3–4 h. After completion reaction monitored by TLC the solvent was evaporated to afford crude products ( 1–15 ). The crude products were recrystallized in methanol to afford 2-(2-bromophenyl) thiazole-5-carbohydrazide-based thioureas ( 1–15 ) (Scheme 1 ) the reaction confirmed by using 1 HNMR 13 CNMR and Mass spectrometry. The absence of NH 2 peak around 4–5 ppm a long with additional peaks of aromatic protons as well as NH peaks in all proton spectra confirm the formation of all desired compounds, the supplementary file of spectra is attached. Biological Activities In recently many studies are carried out for the inhibition of α-amylase and α-glucosidase for the development of potent inhibitors 18 – 22 . All the synthesized scaffolds (1–15) screened for their α-amylase and α-glucosidase inhibitory potential (Table 1 ). Enzyme inhibitory activity assay results The scaffold ( 11) showed reduced inhibitory effect against each enzyme compared to acarbose. When the hydrogen of phenyl ring replaced with fluorine the scaffolds ( 1–3) showed good activity this may be due to its electronegativity of fluorine. When the hydrogen of phenyl ring replaced with chlorine the scaffolds ( 12 and 13) showed good activity this may be due to high electronegative of chlorine. When the hydrogen of phenyl ring replaced with bromine. The brominated scaffolds 4–6 showed good to moderate activity this may be due to big size as well as reduced electronegativity of bromine. When the hydrogen of phenyl ring replaced with methoxy dramatically the scaffolds ( 7–9 ) showed good activity this may be due to oxygen of methoxy attached to phenyl ring. When the hydrogen of phenyl ring replaced with methyl scaffold ( 10 ) showed comparable activity with standard. When the hydrogen of phenyl ring replaced with NO 2 the scaffolds ( 14 and 15) showed moderate activity. Structure Activity Relationship α-Amylase Inhibitory Potential All the synthesized scaffolds as shown in Fig. 1 ( 1–15 ) displayed good to moderate α-amylase inhibitory potential having IC 50 values ranging from 3.10 ± 0.10 µM to 15.40 ± 0.50 µM as compared to standard acarbose (IC 50 =10.30 ± 0.20 µM) (Table 1 ). The structure activity relationship (SAR) was recognized for all scaffolds ( 1–15 ), which displays that the inhibitory potential for the synthesized scaffolds varied due to the numbers, positioned and nature of substituent/s around the phenyl ring. Comparing scaffolds ( 1 , 2 and 3 ) having IC 50 values 3.10 ± 0.10, 4.50 ± 0.20 and 4.20 ± 0.10 µM respectively. These three scaffolds are fluorinated showing more potency than rest of this series which might be due to high electronegativity and small size of fluorine atom. Among the fluorinated scaffolds the ortho substituted scaffold is more potent than meta and para scaffolds as fellow ortho > meta > para. The scaffolds ( 12 and 13 ) having chlorine as a substituent, if they are compared with scaffolds 2 and 3 having fluorine. There is slight decrease in the potency of scaffolds 12 (6.60 ± 0.10 µM) and 13 (4.70 ± 0.10 µM) observed due to less electronegative nature of chlorine. The para substituted scaffold is more than meta substituted scaffold para > meta. The fluorine and chlorine scaffolds ( 1 , 2 , 3, 12 and 13 ) compare with bromine scaffolds ( 4, 5 and 6 ) having IC 50 values 10.40 ± 0.30 µM, 13.60 ± 0.30 µM and 9.30 ± 0.30 µM respectively. They showed weak activity. Greater decline was observed in inhibitory potential of compounds having bromine because of less electronegative and big size of bromo 23 . The para bromo substituted scaffold 6 (IC 50 value = 9.30 ± 0.30 µ M) is more active than the ortho and meta bromo substituted scaffolds 4 and 5 (IC 50 value = 10.40 ± 0.30 µM, 13.60 ± 0.30 µM) para > ortho > meta. The scaffolds having methoxy group as a substituent are 7 , 8 and 9. The IC 50 values of these scaffolds are 7.40 ± 0.20 µM, 9.70 ± 0.30 µM and 5.60 ± 0.20 µM respectively. The substituted methoxy scaffold follow activity pattern para > ortho > meta. If we compare the methyl substituted compound 10 with methoxy scaffolds ( 7–9 ) we found that that is less active than methoxy compounds this may be due to very less election withdrawing power of methyl If nitro substituted scaffolds 14 (15.40 ± 0.50 µM) and 15 (12.20 ± 0.50µM) are compared with chloro substituted scaffolds ( 12 and 13 ). The scaffolds ( 14 and 15 ) become much less potent even than the standard is only due to nature of nitro group which not forming good interaction enzyme. α-Glucosidase Inhibitory Potential All the synthesized scaffolds ( 1–15 ) showed good to moderate α-glucosidase inhibitory potential with IC 50 values between 3.30 ± 0.10 µM to 16.40 ± 0.60 µM when compared with standard acarbose having IC 50 = 9.80.20 ± 0.20 µM (Table 1 ) 24 . The difference between the inhibitory potential of the synthesized scaffolds was studied by structure activity relationship, which depend upon the numbers, nature and positions of substituent/s on the phenyl ring 25 . The scaffolds ( 1 , 2 and 3 ) in this series are fluorinated thiazole-based scaffolds having IC 50 values 3.30 ± 0.10 µM, 4.70 ± 0.20 µM and 5.10 ± 0.20 µM respectively. If we compared these three scaffolds with standard, then all these three scaffolds are more potent than standard. The high electronegativity and small size of fluorine is responsible for their more potency. The ortho (3.30 ± 0.10 µM) substituted scaffold is more potent than the meta (4.70 ± 0.20 µM) and para (5.10 ± 0.20 µM) substituted scaffolds. The scaffolds ( 12 and 13) are chlorinated scaffolds of this series having IC 50 values 7.20 ± 0.30 µM and 4.9 ± 0.20 µM respectively. If both chlorinated scaffolds compared with one another, then the para substituted scaffold is more potent than meta substituted scaffold (Fig. 1 ). Although both scaffolds are more potent than standard. If we compare with fluorinated there is slight decrease in activity this may be due to the decrease in electronegativity. Comparing scaffolds ( 4 , 5 and 6 ) having IC 50 values 11.30 ± 0.30 µM, 13.90 ± 0.40 µM and 9.50 ± 0.30 µM respectively with fluorinated scaffolds ( 1 , 2 and 3 ) and scaffolds ( 12 and 13) having chlrine. There is large decrease in their activity take place. The para bromo substituted scaffolds (9.50 ± 0.30 µM) is more potent even than standard but the ortho and meta bromo substituted scaffolds less potent than standard If the bromine is replaced by methoxy group as in scaffolds ( 7 , 8 and 9 ) having IC 50 values7.90 ± 0.30 µM, 9.60 ± 0.30 µM and 5.70 ± 0.20 µM respectively. All these three scaffolds having less IC 50 values than standard (9.80 ± 0.20 µM). Among the methoxy substituted scaffolds, the para substituted scaffold is more potent than the ortho and meta substituted scaffolds. If we compare the potency of para methyl substituted scaffold ( 10 ) the activity decreases this may be due to the low electronegativity of methyl as compared to methoxy If chlorine in the above scaffolds is replaced by nitro group as in scaffolds ( 14 and 15 ) having IC 50 values 16.40 ± 0.60 µM and 12.80 ± 0.60 µM respectively. Due to nature of nitro group attached at scaffolds ( 14 and 15 ), there is huge decease in their potency occur even though both scaffolds ( 14 and 15 ) become less potent than standard. Molecular docking studies To rationalize the structure-activity relationship (SAR) of the most active compounds, molecular docking studies were performed against the α-amylase active site to understand why different substituents and their positions on the phenyl ring led to varied inhibitory potencies. Prior to the docking simulation, a validation step was carried out on the α-amylase crystal structure (PDB ID: 4W93) to ensure the reliability and accuracy of the docking protocol for this study. The native co-crystallized ligand, Montbretin A, was extracted from the active site and then re-docked using the same parameters and grid box settings that were established for the synthesized compounds. The accuracy of the procedure was confirmed by calculating the Root Mean Square Deviation (RMSD) between the top-ranked predicted binding pose of Montbretin A and its original experimental pose within the crystal structure. The resulting low RMSD value of less than 2.0 Å (1.7 Å) indicated that the chosen docking method successfully reproduced the native binding mode, thereby validating the protocol suitability for predicting the binding interactions of the synthesized thiazole inhibitors. The most potent inhibitors ( 1 , 2 , and 3 ) are all fluorinated, yet they exhibit distinct binding modes that appear to explain their rank order of activity (Fig. 2 ). Compound 1 ( ortho -fluoro), the most active analog (IC₅₀ = 3.10 µM), binds primarily through extensive hydrophobic interactions, with its bromobenzene and thiazole moieties establishing multiple alkyl and π-alkyl contacts with key residues like Ile235, Tyr151, and Ala307. In contrast, compound 3 ( para -fluoro), another highly potent analog (IC₅₀ = 4.20 µM), utilizes a different strategy dominated by a network of conventional hydrogen bonds with Tyr151, Thr163, and Ile148, alongside electrostatic interactions. Compound 2 ( meta -fluoro) also relies on hydrophobic contacts like compound 1 but is distinguished by a unique π- sulfur interaction between its thiourea linkage and the residue Trp58. The docking results suggest that the superior activity of the fluorinated analogs is due to their ability to effectively engage the enzyme through either extensive hydrophobic contacts or a strong hydrogen-bonding network, with the specific position of the fluorine atom dictating which of these binding modes is favored and correlating with their inhibitory potentials. As for less potent analogs such as compounds 6 and 9 , a more detailed docking analysis of these compounds clarifies their moderate potency in comparison to the leading fluorinated analogs (Fig. 3 ). It observed that the para -bromo substituted compound 6 (IC₅₀ = 9.30 µM) and the para -methoxy substituted compound 9 (IC₅₀ = 5.60 µM) could possibly form hydrogen bonds with residues like His305 and Asp300, respectively, in addition to hydrophobic interactions. Compound 6 , with a para -bromo substituent, binds itself firmly through various interactions. It forms two crucial hydrogen bonds between its carbonyl oxygen and the His305 sidechain at a distance of 2.09Å, and another between its thiourea group and Asp300 at the distance of 2.99Å. The compound is further stabilized by electrostatic forces, including a π-cation interaction with His201 (3.02Å) and a π-anion interaction with Asp356 (4.15Å), alongside extensive hydrophobic contacts. Notably, its para -bromo substituent actively participates in binding through close alkyl interactions with Lys200 (3.91Å) and Ile235 (4.05Å). In a similar fashion, compound 9 , the most potent methoxy-substituted analog (IC₅₀ = 5.60 µM), formed a conventional hydrogen bond between its carbonyl oxygen and Asp300 at 2.40Å. The analysis highlights a stabilizing halogen bond between the bromine on the compound main ring with the backbone oxygen from Ile235 at 3.36Å. Its fit is reinforced by numerous hydrophobic contacts, particularly the π-alkyl interactions between its para -methoxy group and the sidechains of Trp59 (at 3.84Å and 4.81Å). While both compounds 6 and 9 form multiple significant bonds, the specific nature and geometry of these interactions result in good inhibitory activity. This detailed analysis clarifies that their binding modes, while effective, are less optimal than those of the more potent fluorinated analogs, explaining their position in the overall Materials and Methods All solvents and chemicals used in synthesis purchased from Sigma Aldrich and used without further purification. Analytical grade solvents used for reaction as well as washing. NMR spectra recorded on JEOL spectrometer at 500 MHz and 125 MHz as internal standard TMS used and DMSO-d 6 used as solvent. For HR-EIMS high-resolution mass spectra (electron impact, 60 eV) on a Finnigan MAT-311 A instrument (Germany) were used. For visualization of chromatogram, a UV lamp (Schimazdu, Germany) of wavelength 254/365 used. General procedure for the synthesis of thiazole-based scaffolds Thiazole-thiourea analogs synthesized by treating 2-(2-bromophenyl) thiazole-5-carbohydrazide (1 mmol) with numerous aryl isothiocyanates (1 mmol) in the chloroform (10 mL). The reaction mixture kept on stirring for 3–4 h. The reaction completion was carefully monitored using thin layer chromatography (TLC). After accomplishment of reaction the solvent evaporated by vacuum rotatory evaporator to afford crude products 1–15 . The crude products were recrystallized in methanol to afford 2-(2-bromophenyl) thiazole-5-carbohydrazide-based thiourea ( 1–15 ). All synthesized compounds fully characterized by using different spectroscopic methods, 1 HNMR, 13 CNMR, HREI MS. Characterization of thiazole-based scaffolds 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(2-fluorophenyl)hydrazine-1-carbothioamide ( 1 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.74 (s, 1H, NH), 9.97 (s, 1H, NH), 9.90 (s, 1H, NH), 8.51 (s, 1H, Ar), 8.45 (d, J = 1.8 Hz, 1H, Art), 8.05 (d, J = 6.5 Hz, 1H, Ar), 7.75 (d, J = 6.7 Hz, 1H, Ar), 7.51(t, J = 6.5 Hz, 2H, Ar), 7.32 (m, 2H, Ar), 6.98 (t, J = 7.8 Hz, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 213.3, 180.7, 165.1 (d, J = 153, C-F), 148.8, 141.0, 141.0, 134.4, 133.3, 131.4, 129.6, 128.6, 126.0, 125.6, 123.2, 122.6, 119.5, 113.8; HREI-MS: m/z calculated for C 17 H 12 BrFN 4 OS 2 [M] + 449.9620; found: 449.9592. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(3-fluorophenyl)hydrazine-1-carbothioamide ( 2 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.9 (s, 1H, Ar), 10.05 (s, 1H, NH), 9.85 (s, 1H, NH), 8.65 (d, J = 6.5Hz, 1H, Ar), 8.25(d, J = 1.5Hz, 1H, Ar), 7.38 (d, J = 4.5 Hz, 1H, Ar), 7.22(d, J = 2.0 Hz, 1H, Ar), 7.13(s, 1H, Ar), 7.08 (s, 1H, Ar), 7.02 (m, 2H, Ar), 6.88 (s, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 187.4, 182.1, 166.2 (d, J = 133, C-F), 160.1, 149.0, 134.4, 133.3, 131.4, 130.5, 128.7, 128.0, 125.9, 125.6, 124.0, 122.6, 115.6, 115.5; HREI-MS: m/z calculated for C 17 H 12 BrFN 4 OS 2 [M] + 449.9620; found: 449.9599. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(4-fluorophenyl)hydrazine-1-carbothioamide ( 3 ) 1 HMNR (500 MHz, DMSO-d 6 ); δ 10.81 (s, 1H, NH), 10.17 (s, 1H, NH), 9.85 (s, 1H, NH), 8.53 (s, 1H, Ar), 8.49 (m, 2H, Ar), 8.06 (dd, J = 6.0, 2.0 Hz, 2H, Ar), 8.01 (dd, J = 6.6, 2.5 Hz, 1H, Ar), 7.52 (dd, J = 6.6, 2.5 Hz, 1H, Ar), 7.63 (t, J = 6.8, 1H, Ar), 7.52 (t, J = 6.6, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 180.9, 171.0, 165.2 (d, J = 158, C-F), 160.1, 155.7, 148.7, 147.1, 140.5, 134.3, 133.3, 131.4, 129.0, 128.6, 126.1, 125.6, 122.6, 119.5; HREI-MS: m/z calculated for C 17 H 12 BrFN 4 OS 2 [M] + 449.9620; found: 449.9582. N -(2-Bromophenyl)-2-(2-(2-Bromophenyl)thiazole-5-carbonyl)hydrazine-1-carbothioamide ( 4 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ δ 10.82 (s, 1H, NH), 10.53 (s, 1H, NH), 10.24 (s, 1H, NH), 8.52 (s, 1H, Ar), 8.45 (s, 1H, Ar), 8.22 (dd, J = 7.6, 2.0 Hz, 2H, Ar), 8.06 (dd, J = 7.6, 2.1 Hz, 1H,Ar), 7.94 (d, J = 7.7 Hz, 2H,Ar), 7.75 (d, J = 6.6 Hz, 1H,Ar), 7.49 (t, J = 6.5, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ): δ 180.6, 178.4, 148.8, 146.3, 143.2, 134.3, 133.3, 132.5, 131.4, 130.2, 128.6, 126.2, 125.6, 124.7, 123.5, 122.6, 120.1; HREI-MS: m/z calculated for C 17 H 12 Br 2 N 4 OS 2 [M] + 509.8819; found: 509.8766. N -(3-Bromophenyl)-2-(2-(2-Bromophenyl)thiazole-5-carbonyl)hydrazine-1-carbothioamide ( 5 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.75 (s, 1H, NH), 9.99 (s, 1H, NH), 9.95 (s, 1H, NH), 8.51 (s, 1H, Ar), 8.46 (s, 1H, Ar), 8.06 (dd, J = 6.5, 2.0 Hz, 1H, Ar), 7.75 (dd, J = 7.6, 2.6 Hz, 2H, Ar), 7.52–7.47 (m, 2H, Ar), 7.37 (t, J = 6.7, 1H, Ar), 7.21 (dd, J = 6.8, 2.3 Hz, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ180.7, 165.1, 163.6, 151.1, 148.9, 140.7, 134.3, 132.0, 133.3, 131.4, 129.3, 128.6, 126.0, 125.6, 125.6, 122.6, 120.1; HREI-MS: m/z calculated for C 17 H 12 Br 2 N 4 OS 2 [M] + 509.8819; found: 509.8773. N -(4-Bromophenyl)-2-(2-(2-Bromophenyl)thiazole-5-carbonyl)hydrazine-1-carbothioamide ( 6 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.72 (s, 1H, Ar), 9.91 (s, 1H, NH), 9.83 (s, 1H, NH), 8.49 (s, 1H Ar), 8.45 (s, 1H Ar), 8.39 (d, J = 3.0 Hz, 1H, Ar), 8.32 (s, 1H Ar), 8.06 (dd, J = 7.7, 2.0 Hz, 1H, Ar), 8.02 (dd, J = 6.5, 2.0 Hz, 1H, Ar), 7.75 (t, J = 7.6, 1H, Ar), 7.52–7.44 (m, 1H, Ar), 7.17 (t, J = 7.3, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 180.8, 165.2, 159.6, 149.6, 139.8, 134.5, 133.3, 133.2, 131.4, 131.3, 128.6, 125.9, 125.6, 125.6, 124.0, 122.6, 122.5; HREI-MS: m/z calculated for C 17 H 12 Br 2 N 4 OS 2 [M] + 509.8819; found: 509.8781. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(2-methoxyphenyl)hydrazine-1-carbothioamide ( 7 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.68 (s, 1H, NH), 9.91 (s, 1H, NH), 9.71 (s, 1H, NH), 8.49 (s, 1H Ar), 8.46 (d, J = 3.2 Hz, 2H, Ar), 8.06 (dd, J = 6.5, 2.3 Hz, 1H, Ar), 7.75 (dd, J = 6.7, 2.5 Hz, 1H, Ar), 7.52 (t, J = 6.6, 1H, Ar), 7.31(d, J = 7.0 Hz, 2H, Ar), 6.90 (d, J = 7.5 Hz, 2H, Ar), 3.74 (s, 3H, OCH 3 ). 13 CNMR (125 MHz, DMSO-d 6 ); δ 180.6, 165.0, 161.3, 156.6, 149.1, 140.2, 134.4, 133.3, 132.0, 131.4, 131.3, 128.6, 125.8, 125.6, 122.6, 120.1, 113.1, 55.4; HREI-MS: m/z calculated for C 18 H 15 BrN 4 O 2 S 2 [M] + 461.9820; found: 461.9761. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(3-methoxyphenyl)hydrazine-1-carbothioamide ( 8 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.70 (s, 1H, NH), 9.84 (s, 1H, NH), 9.71 (s, 1H, NH), 8.50 (s, 1H, Ar), 8.45 (s, 1H, Ar), 8.06 (dd, J = 6.5, 2.2 Hz, 1H, Ar), 7.74 (dd, J = 6.6, 2.5 Hz, 1H, Ar), 7.52 (t, J = 6.6, 1H, Ar), 7.24 (t, J = 6.7, 2H, Ar), 7.09 (dd, J = 6.7, 2.3 Hz, 1H, Ar), 6.74 (dd, J = 6.9, 2.2 Hz, 1H, Ar), 3.75 (s, 3H, OCH 3 ). 13 CNMR (125 MHz, DMSO-d 6 ); δ 181.4, 165.1, 158.8, 149.1, 144.2, 140.3, 134.4, 133.3, 131.4, 130.5, 130.2, 129.8, 128.6, 128.4, 125.9, 125.6, 122.6, 55.0; HREI-MS: m/z calculated for C 18 H 15 BrN 4 O 2 S 2 [M] + 461.9820; found: 461.9777. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(4-methoxyphenyl)hydrazine-1-carbothioamide ( 9 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.89 (s, 1H, NH), 10.05 (s, 1H, NH), 9.85 (s, 1H, Ar), 9.24 (s, 1H, Ar), 8.52 (s, 1H, Ar), 8.45 (s, 1H, Ar), 8.06 (dd, J = 6.5, 2.2 Hz, 1H, Ar), 7.75 (dd, J = 6.7, 2.4 Hz, 1H, Ar), 7.51 (t, J = 6.6 Hz, 1H, Ar), 7.34 (t, J = 6.2 Hz, 1H, Ar), 7.18–7.15 (m, 1H, Ar), 6.93 (t, J = 6.4, 1H, Ar), 3.73 (s, 3H, OCH 3 ). 13 CNMR (125 MHz, DMSO-d 6 ); δ 181.0, 165.1, 155.5, 148.7, 134.4, 133.4, 131.4, 129.1, 128.7, 126.1, 125.6, 125.5, 122.6, 120.8, 119.7, 112.3, 111.4, 55.6; HREI-MS: m/z calculated for C 18 H 15 BrN 4 O 2 S 2 [M] + 461.9820; found: 461.9752. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(p-tolyl)hydrazine-1-carbothioamide ( 10 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.69 (s, 1H, NH), 9.91 (s, 1H, NH), 9.75 (s, 1H, NH), 8.49 (s, 1H Ar), 8.40 (d, J = 2.0 Hz, 1H, Ar), 8.06 (dd, J = 6.5, 2.3 Hz, 1H, Ar), 8.01 (d, J = 2.3 Hz, 1H, Ar), 7.74–7.71 (m, 1H, Ar), 7.52–7.47 (m, 2H, Ar), 7.13 (d, J = 6.9 Hz, 1H, Ar), 2.28 (s, 3H, CH 3 ). 13 CNMR (125 MHz, DMSO-d 6 ); δ 181.3, 165.2, 159.6, 149.6, 134.5, 134.4, 133.3, 133.2, 131.4, 131.3, 128.7, 125.8, 125.6, 125.6, 124.0, 122.6, 122.5, 20.5; HREI-MS: m/z calculated for C 18 H 15 BrN 4 OS 2 [M] + 445.9871; found: 445.9844. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -phenylhydrazine-1-carbothioamide ( 11 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.74 (s, 1H, NH), 9.91 (s, 1H, NH), 9.87 (s, 1H, NH), 8.50 (s, 1H Ar), 8.45 (d, J = 2.0 Hz, 1H, Ar), 8.39 (d, J = 7.5 Hz, 1H, Ar), 8.32 (m, 1H, Ar), 8.06 (d, J = 2.2 Hz, 1H, Ar), 8.02 (d, J = 7.0 Hz, 1H, Ar), 7.73 (dd, J = 7.5, 2.0 Hz, 1H, Ar), 7.49 (t, J = 6.6, 2H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 180.3, 165.2, 159.6, 149.6, 134.5, 133.2, 132.7, 131.3, 130.2, 128.6, 128.5, 128.5, 126.4, 125.6, 125.6, 124.0, 122.5; HREI-MS: m/z calculated for C 17 H 13 BrN 4 OS 2 [M] + 431.9714; found: 431.9667. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(3-chlorophenyl)hydrazine-1-carbothioamide ( 12 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.78 (s, 1H, NH), 9.93 (s, 1H, NH), 9.67 (s, 1H, NH), 8.51 (s, 1H Ar), 8.47 (s,1H, Ar), 8.06 (dd, J = 6.5, 2.2 Hz, 1H, Ar), 7.74 (dd, J = 6.6, 2.5 Hz, 1H, Ar), 7.65 (d, J = 6.0 Hz, 1H, Ar), 7.52 (t, J = 6.5 Hz, 1H, Ar), 7.42–7.37 (m, 2H, Ar), 7.21(t, J = 7.6 Hz, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 181.0, 165.0, 160.1, 149.0, 134.4, 133.3, 132.3, 132.0, 130.2, 129.8, 131.4, 128.7, 127.6, 127.6, 126.0, 125.6, 122.6; HREI-MS: m/z calculated for C 17 H 12 BrClN 4 OS 2 [M] + 465.9324; found: 465.9301. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(4-chlorophenyl)hydrazine-1-carbothioamide ( 13 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.74 (s, 1H, NH), 9.99 (s, 1H, NH), 9.90 (s, 1H, NH), 8.51 (s, 1H Ar), 8.46 (s,1H, Ar), 8.06 (dd, J = 6.5, 2.1 Hz, 1H, Ar), 7.75 (dd, J = 6.7, 2.4 Hz, 2H, Ar), 7.56 (d, J = 6.9 Hz, 1H, Ar), 7.51 (t, J = 6.5 Hz, 1H, Ar), 7.35 (dd, J = 6.9, 1.3 Hz, 1H, Ar), 7.29 (t, J = 6.6 Hz, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 170.8, 169.4, 165.2, 148.9, 147.1, 140.9, 140.5, 140.5, 134.4, 133.4, 132.2, 131.4, 128.6, 128.6, 125.7, 125.6, 122.6; HREI-MS: m/z calculated for C 17 H 12 BrClN 4 OS 2 [M] + 465.9324; found: 465.9295. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(3-nitrophenyl)hydrazine-1-carbothioamide ( 14 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.73(s, 1H, NH), 9.92 (s, 1H, NH), 9.88 (s, 1H, NH), 8.57 (s, 1H, Ar), 8.50 (s, 1H, Ar), 8.32 (d, J = 3.3 Hz, 1H, Ar), 8.06 (dd, J = 6.5, 2.2 Hz, 1H, Ar), 7.74 (dd, J = 6.6, 2.5 Hz, 2H, Ar), 7.52 (t, J = 6.6 Hz, 1H, Ar), 7.39 (d J = 8.2, 2.7 Hz, 2H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 181.7, 165.1, 149.0, 144.2, 143.5, 138.2, 134.3, 133.3, 131.4, 129.5., 129.2, 128.6, 127.7, 125.9, 125.6, 122.6, 120.3; HREI-MS: m/z calculated for C 17 H 12 BrN 5 O 3 S 2 [M] + 476.9565; found: 476.9546. 2-(2-(2-Bromophenyl)thiazole-5-carbonyl)- N -(4-nitrophenyl)hydrazine-1-carbothioamide ( 15 ) 1 HNMR (500 MHz, DMSO-d 6 ); δ 10.71 (s, 1H, NH), 9.91 (s, 1H, NH), 9.82 (s, 1H, NH), 8.50 (s, Ar), 8.46 (d, J = 1.5Hz, 1H, Ar), 8.05 (dd, J = 6.5, 2.3Hz, 1H, Ar), 7.74 (dd, J = 7.4, 2.3 Hz, 1H, Ar), 7.52–7.46 (m, 2H, Ar), 7.33 (t, J = 6.8 Hz, 2H, Ar), 7.15 (t, J = 6.0 Hz, 1H, Ar). 13 CNMR (125 MHz, DMSO-d 6 ); δ 180.2, 165.1, 149.1, 148.7, 148.7, 139.2, 134.4, 133.4, 131.4, 128.6, 127.8, 125.9, 125.6, 124.4, 123.1, 123.1, 122.6; HREI-MS: m/z calculated for C 17 H 12 BrN 5 O 3 S 2 [M] + 476.9565; found: 476.9533. Biological activity α-Amlyase and α-glucosidase inhibition assay inhibitory activity Enzyme and reagents purchased from sigma Aldrich, we used the same protocol reported in our previous paper 23 , 25 – 27 for detail kindly see supplementary data. Molecular Docking In this study AutoDock 4.2 28 was utilized to dock the small molecules into the active site of the protein. A homology model of α-glucosidase that was constructed based on the crystal structure of isomaltase from Saccharomyces cerevisiae (PDB ID: 3A4A) using SwissModel 29 . While for α-amylase enzyme, the crystal structure (PDB ID: 4W93) was downloaded from protein databank website. The structure of α-amylase enzyme was optimized by removing water molecules, adding hydrogen atoms, adding charge, and repairing end residues. For α-glucosidase enzyme, the grid map was generated based on 0.375 Å spacing between grid points, and the center of the grid box was placed at coordinate x = 14.885107, y = -11.366557, and z = 18.413239. The dimensions of the active site box were set at 60 × 60 × 60 Å. While for α-amylase enzyme, the grid map was generated using 0.375 Å spacing between grid points, and the center of the grid box was placed at coordinate x = -9.632488, y = 4.340907, and z = -23.107256. Each docked system was performed with 150 runs using Lamarckian genetic algorithm function. Protein‒ligand interactions were visualized and analyzed using Discovery Studio Visualizer 3.5. Conclusion In this work fifteen thiazole-based scaffolds ( 1 – 15 ) have been synthesized and characterized using various spectroscopic techniques including 1 HNMR, 13 CNMR, HREI MS. Then all these synthesized scaffolds were screened for their alpha amylase and alpha glucosidase inhibitory activity. All synthesized thiazole-based scaffolds displayed wide range of inhibition for both targeted enzymes. The IC 50 values for alpha amylase and alpha glucosidase are 3.10 ± 0.10 µM to 15.40 ± 0.50 µM and 3.10 ± 0.10 µM to 15.40 ± 0.50 µM respectively. The scaffolds ( 1 , 2 and 3 ) have been found most potent in the synthesized series of thiazole-based scaffolds. These are fluorinated thiazole-based scaffolds in which the fluorine might be responsible for their potency. The binding interactions between most potent scaffold and targeted enzymes have been studied through molecular docking, which confirmed the binding interaction of compounds with enzymes’ active site. Declarations Funding: This research did not receive funding. Author Contribution M. T and F. R (Synthesis); M. A and S. A. A. S. (Characterization of compounds); N. U. and F. S. A (Purification of compounds); A. S. and K. M. K. (Bioassay); S. I. (Molecular docking); M. T, M. A. and K. M. K. Writing and editing manuscript Acknowledgement Authors would like to thank Imam Abdulrahman Bin Faisal University for providing excellent lab facilities. Data Availability The datasets generated and/or analyzed during the current study are not publicly available because they are private, but are available from the corresponding author on reasonable request. References Wondafrash, D. Z. et al. Potential effect of hydroxychloroquine in diabetes mellitus: a systematic review on preclinical and clinical trial studies. J. Diabetes Res. 5214751 (2020). (2020). Evans-Molina, C. et al. The heterogeneity of type 1 diabetes: implications for pathogenesis, prevention, and treatment—2024 Diabetes, Diabetes Care, and Diabetologia Expert Forum. Diabetes Care , dci250013 (2025). Dludla, P. V. et al. Pancreatic β-cell dysfunction in type 2 diabetes: Implications of inflammation and oxidative stress. World J. Diabetes . 14 , 130 (2023). Dirir, A. M., Daou, M., Yousef, A. F. & Yousef, L. F. 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Synthesis of thiazole-based-thiourea analogs: as anticancer, antiglycation and antioxidant agents, structure activity relationship analysis and docking study. J. Biomol. Struct. Dyn. 41 , 12077–12092 (2023). Naz, H. et al. Investigation of novel benzimidazole-based indole/thiazole hybrids derivatives as effective anti-diabetics and anti-alzheimer's agents: Structure-activity relationship insight, in vitro and in silico approaches. J. Mol. Struct. 1312 , 138592 (2024). Taha, M. et al. Exploring thiazole-based Schiff base analogs as potent α-glucosidase and α-amylase inhibitor: their synthesis and in-silico study. J. Mol. Struct. 1287 , 135672 (2023). Hussain, R. et al. Multipotent cholinesterase inhibitors for the treatment of Alzheimer’s disease: synthesis, biological analysis and molecular docking study of benzimidazole-based thiazole derivatives. Molecules 27 , 6087 (2022). El-Hussieny, M. et al. Iron (III) triflate, a new efficient catalyst for the synthesis of new bis (oxy)-bis (phenylene)-ethane-Schiff bases: In vitro antioxidant, anti-inflammatory, α-Amylase, α-Glucosidase biochemical screening, and in silico study. J Mol. Struct , 143627 (2025). Nguyen, C. Q. et al. Pharmacoinformatics model based on the para-aminochalcone core to screening dual alpha-amylase/alpha-glucosidase and β-TC-6 inhibitors: Combining experimental and machine learning for diabetes drug discovery. J Mol. Struct , 143193 (2025). Sireesha, S. M., Prasad, B. D. & Design Synthesis, and Antidiabetic Evaluation of N-2-Aryl-1, 2, 3-Triazoles (NATs) as Potent α-Amylase and α-Glucosidase Inhibitors: A Computational and Biochemical Investigation. J Mol. Struct , 142771 (2025). Abdelgawad, M. A. et al. Synthesis and characterization of novel pyrazoline derivatives as dual α-amylase/α-glucosidase inhibitors: molecular modeling and kinetic study. J. Mol. Struct. 1339 , 142350 (2025). Kucuk, C., Celik, S., Yurdakul, S. & Coteli, E. A new Ag (I)-complex of 5-chloroquinolin-8-ol ligand: Synthesis, spectroscopic characterization, and DFT investigations, in vitro antioxidant (DPPH and ABTS), α-glucosidase, α-amylase inhibitory activities with protein-binding analysis. J. Mol. Struct. 1325 , 141285 (2025). Taha, M. et al. Synthesis, in vitro evaluation and molecular docking studies of hybrid 4-quinolinyl bearing 1, 3, 4-thiadiazole-2-amine as a new inhibitor of α-amylase and α-glucosidase. J. Mol. Struct. 1282 , 135173 (2023). Mukhliss, L. et al. Synthesis and novel structural hybrid analogs of oxindole derivatives bearing piperidine ring, their antidiabetics II activity and molecular docking study. J. Mol. Struct. 1332 , 141666 (2025). Saleem, F. et al. Bioevaluation of synthetic pyridones as dual inhibitors of α-amylase and α‐glucosidase enzymes and potential antioxidants. Arch. Pharm. 356 , 2200400 (2023). Javid, M. T. et al. Synthesis, in vitro α-glucosidase inhibitory potential and molecular docking study of thiadiazole analogs. Bioorg. Chem. 78 , 201–209 (2018). Rahim, F. et al. Triazinoindole analogs as potent inhibitors of α-glucosidase: Synthesis, biological evaluation and molecular docking studies. Bioorg. Chem. 58 , 81–87 (2015). Morris, G. M. et al. AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility. JCoCh 30 , 2785–2791 (2009). Waterhouse, A. et al. SWISS-MODEL: homology modelling of protein structures and complexes. NAR 46, W296-W303 (2018). Table 1 Table 1 is available in the Supplementary Files section. Schemes Scheme 1 is available in the Supplementary Files section Additional Declarations No competing interests reported. Supplementary Files Supplementarydata.pdf Scheme1.docx Table1.docx Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 14 Feb, 2026 Reviewers agreed at journal 11 Feb, 2026 Reviewers invited by journal 11 Feb, 2026 Editor invited by journal 28 Jan, 2026 Editor assigned by journal 24 Jan, 2026 Submission checks completed at journal 24 Jan, 2026 First submitted to journal 14 Jan, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8601560","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":591440307,"identity":"b68e34f2-351a-41cc-ab4a-a0bd507e9d58","order_by":0,"name":"Muhammad Taha","email":"","orcid":"","institution":"Imam Abdulrahman Bin Faisal University","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Taha","suffix":""},{"id":591440308,"identity":"2faf7fb0-aabd-4faa-92bc-a0663d80704a","order_by":1,"name":"Munther Alomari","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAklEQVRIiWNgGAWjYJACZjDJDiYl5BgYEgjqYGxG0ihhTLIWhsQGQlp0Z+Qef1xQcY+Bv5n56IafeyzS+9tzDBh+1DDI8+PQYnYjL7F5xpliBonDbGk3e55J5M4488aAsecYg+HMBlxacgybeduAbjnMY3aD54BEbsMNoC28DQyMGw7g0/IvgUEeqOXmnwMS6fJALYx/Gxjs9+PV0pDAYADUchtoS4IBUAsz0JbEDbj8cuaN4WyeYwk8hkC/3JY5IGG48cyzgsMyxySSZ+Cy5XiOwWeemgQ5uePNx26+OVAnL3c8eePDNzU2tv04vA8DPCg8oPkS+NWPglEwCkbBKMALAJAlWyKbKrupAAAAAElFTkSuQmCC","orcid":"","institution":"Abu Dhabi University","correspondingAuthor":true,"prefix":"","firstName":"Munther","middleName":"","lastName":"Alomari","suffix":""},{"id":591440309,"identity":"20906adc-6a42-465a-80e4-058d99c50913","order_by":2,"name":"Faisal S. 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2","display":"","copyAsset":false,"role":"figure","size":127157,"visible":true,"origin":"","legend":"\u003cp\u003eThe 2D-interaction diagram for compounds \u003cstrong\u003e1, 2, \u003c/strong\u003eand\u003cstrong\u003e 3\u003c/strong\u003e against \u003cem\u003eα\u003c/em\u003e-amylase.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8601560/v1/263b68f8e2687534eb8630a0.png"},{"id":103049330,"identity":"5416897c-5d0b-4dcc-972d-bb706e7c3428","added_by":"auto","created_at":"2026-02-20 07:39:53","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":182899,"visible":true,"origin":"","legend":"\u003cp\u003eThe 2D interaction diagram for compounds \u003cstrong\u003e7, 8, \u003c/strong\u003eand\u003cstrong\u003e 9\u003c/strong\u003e against \u003cem\u003eα\u003c/em\u003e-amylase.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8601560/v1/38b50d1813b0425f7bc2a048.png"},{"id":103050909,"identity":"fd314333-9201-415e-8c80-e94f213ac92d","added_by":"auto","created_at":"2026-02-20 07:56:53","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1228279,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8601560/v1/acb13cd6-a8fb-46da-b761-01d721f42686.pdf"},{"id":102771980,"identity":"f2f42cf0-6b05-4dc2-a159-906af2d25ed4","added_by":"auto","created_at":"2026-02-16 12:50:21","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1131153,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarydata.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8601560/v1/4585696038096c4d781bcff1.pdf"},{"id":102771977,"identity":"da2b6bea-3f73-4cb7-ab92-aea36f596079","added_by":"auto","created_at":"2026-02-16 12:50:20","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":26750,"visible":true,"origin":"","legend":"","description":"","filename":"Scheme1.docx","url":"https://assets-eu.researchsquare.com/files/rs-8601560/v1/e1d4795d708f87ce6f382809.docx"},{"id":102771978,"identity":"b9daaaa4-6685-435c-a1ec-fb63b085a89e","added_by":"auto","created_at":"2026-02-16 12:50:21","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":73055,"visible":true,"origin":"","legend":"","description":"","filename":"Table1.docx","url":"https://assets-eu.researchsquare.com/files/rs-8601560/v1/e7a34c9364d571f96a334455.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Identification thiazole-based analogs as potent antidiabetic agents and their molecular docking study","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIn today's world, the development of diabetes mellitus (DM) has been identified as one of the greatest health issues worldwide. In 2011 information collected by WHO there are approximately 366\u0026nbsp;million patients affected from diabetes and by 2030 the affected people increases to 522\u0026nbsp;million worldwide \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Type 1 diabetes is a chronic condition in which the immune system attacks the insulin producing pancreatic beta cells. Thus to maintain the level of the pancreatic beta cells, insulin administered to cure type 1 diabetes \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. While diabetes-II is caused when beta cells of the pancreas secrete insufficient amount of insulin \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe α-amylase is the digestive enzymes that catalyze the hydrolysis of carbohydrate that result in blood sugar. The alpha amylase is secreted by pancreas and salivary gland and is responsible for the hydrolysis of starch and oligosaccharide into sugar. Alpha-glucosidase enzyme, which is responsible for the breakdown of polysaccharides is present on the brush boarder of small intestine \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. The inhibition of these enzymes plays important role in controlling blood glucose by decreasing carbohydrate metabolism \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. Several anti-diabetic drugs are used in clinical practice nowadays such as acarbose, voglibose and miglitol that reduce the activity of α-glucosidase enzyme which then consequently interfere the carbohydrate metabolism and reduces the chance of postprandial hyper-glycaemia and hyper- insulinemia \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. However, the use of these drugs has adverse side effect such as flatulence, abdominal pain, dizziness and diarrhea, because of microorganisms in the colon fermenting excessive carbohydrates. In rare instances, intestinal difficulties such as pneumatosis cystoids intestinalis, liver problems, particularly with acarbose, and allergic skin reactions are less frequent but more dangerous adverse effects \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Therefore, there is intense need to develop more potent and efficient drugs on priority basis having low side effects.\u003c/p\u003e \u003cp\u003eThiazole is a heterocyclic molecule having numerous pharmacological applications such as inflammatory \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e hypertension \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e and bacterial disease \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. Amino-thiazoles are popular as estrogen receptors ligands \u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e and an innovative class of adenosine receptor antagonists \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. Thiazole moiety is present in numerous medications such as in Fanetizole an anti-inflammatory drug, Abafungin an antifungal agent, Famotidine a histamine H2 receptor antagonist, and Pamicogrel an anti-cancer agent \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. The thiazole analogs were reported as antidiabetic and anti-Alzheimer agents \u003csup\u003e\u003cspan additionalcitationids=\"CR15 CR16\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe method of molecular hybridization is an important approach to research and development of new biologically active compounds, which involves the hybridization of two different active substances from the same molecule. Due to the wide range of pharmacological activities associated with thiazoles and thiourea moieties, a type of thiazole-thiourea derivative has been designed and studied for the inhibition of diabetes mellitus. The aim of this study is to identify the best thiazole-thiourea derivatives as inhibitors of type 2 and study interaction of enzymes (α-amylase and α-glucosidase) with active compounds \u003cem\u003evia\u003c/em\u003e molecular docking study.\u003c/p\u003e"},{"header":"Results and discussion","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eChemistry\u003c/h2\u003e \u003cp\u003eFifteen (\u003cb\u003e1\u003c/b\u003e\u0026ndash;\u003cb\u003e15\u003c/b\u003e) thiazoles bearing thiourea analogs were synthesized by reacting 2-(2-bromophenyl) thiazole-5-carbohydrazide (1 mmol) with various aryl isothiocyanates (1 mmol) in chloroform (10 mL) as a solvent and allowing the reaction mixture to proceed for 3\u0026ndash;4 h. After completion reaction monitored by TLC the solvent was evaporated to afford crude products (\u003cb\u003e1\u0026ndash;15\u003c/b\u003e). The crude products were recrystallized in methanol to afford 2-(2-bromophenyl) thiazole-5-carbohydrazide-based thioureas (\u003cb\u003e1\u0026ndash;15\u003c/b\u003e) (Scheme \u003cspan refid=\"Sch1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) the reaction confirmed by using \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003eHNMR \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003eCNMR and Mass spectrometry. The absence of NH\u003csub\u003e2\u003c/sub\u003e peak around 4\u0026ndash;5 ppm a long with additional peaks of aromatic protons as well as NH peaks in all proton spectra confirm the formation of all desired compounds, the supplementary file of spectra is attached.\u003c/p\u003e \n\u003ch3\u003eBiological Activities\u003c/h3\u003e\n\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eIn recently many studies are carried out for the inhibition of α-amylase and α-glucosidase for the development of potent inhibitors \u003csup\u003e\u003cspan additionalcitationids=\"CR19 CR20 CR21\" citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. All the synthesized scaffolds (1\u0026ndash;15) screened for their α-amylase and α-glucosidase inhibitory potential (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e\n\u003ch3\u003eEnzyme inhibitory activity assay results\u003c/h3\u003e\n\u003cp\u003eThe scaffold (\u003cb\u003e11)\u003c/b\u003e showed reduced inhibitory effect against each enzyme compared to acarbose. When the hydrogen of phenyl ring replaced with fluorine the scaffolds (\u003cb\u003e1\u0026ndash;3)\u003c/b\u003e showed good activity this may be due to its electronegativity of fluorine. When the hydrogen of phenyl ring replaced with chlorine the scaffolds (\u003cb\u003e12\u003c/b\u003e and \u003cb\u003e13)\u003c/b\u003e showed good activity this may be due to high electronegative of chlorine. When the hydrogen of phenyl ring replaced with bromine. The brominated scaffolds \u003cb\u003e4\u0026ndash;6\u003c/b\u003e showed good to moderate activity this may be due to big size as well as reduced electronegativity of bromine. When the hydrogen of phenyl ring replaced with methoxy dramatically the scaffolds (\u003cb\u003e7\u0026ndash;9\u003c/b\u003e) showed good activity this may be due to oxygen of methoxy attached to phenyl ring. When the hydrogen of phenyl ring replaced with methyl scaffold (\u003cb\u003e10\u003c/b\u003e) showed comparable activity with standard. When the hydrogen of phenyl ring replaced with NO\u003csub\u003e2\u003c/sub\u003e the scaffolds (\u003cb\u003e14\u003c/b\u003e and \u003cb\u003e15)\u003c/b\u003e showed moderate activity.\u003c/p\u003e\n\u003ch3\u003eStructure Activity Relationship\u003c/h3\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eα-Amylase Inhibitory Potential\u003c/h2\u003e \u003cp\u003eAll the synthesized scaffolds as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e (\u003cb\u003e1\u0026ndash;15\u003c/b\u003e) displayed good to moderate α-amylase inhibitory potential having IC\u003csub\u003e50\u003c/sub\u003e values ranging from 3.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M to 15.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50 \u0026micro;M as compared to standard acarbose (IC\u003csub\u003e50\u003c/sub\u003e=10.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The structure activity relationship (SAR) was recognized for all scaffolds (\u003cb\u003e1\u0026ndash;15\u003c/b\u003e), which displays that the inhibitory potential for the synthesized scaffolds varied due to the numbers, positioned and nature of substituent/s around the phenyl ring.\u003c/p\u003e \u003cp\u003eComparing scaffolds (\u003cb\u003e1\u003c/b\u003e, \u003cb\u003e2\u003c/b\u003e and \u003cb\u003e3\u003c/b\u003e) having IC\u003csub\u003e50\u003c/sub\u003e values 3.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10, 4.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 and 4.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M respectively. These three scaffolds are fluorinated showing more potency than rest of this series which might be due to high electronegativity and small size of fluorine atom. Among the fluorinated scaffolds the \u003cem\u003eortho\u003c/em\u003e substituted scaffold is more potent than \u003cem\u003emeta\u003c/em\u003e and \u003cem\u003epara\u003c/em\u003e scaffolds as fellow ortho\u0026thinsp;\u0026gt;\u0026thinsp;meta\u0026thinsp;\u0026gt;\u0026thinsp;para.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe scaffolds (\u003cb\u003e12\u003c/b\u003e and \u003cb\u003e13\u003c/b\u003e) having chlorine as a substituent, if they are compared with scaffolds \u003cb\u003e2\u003c/b\u003e and \u003cb\u003e3\u003c/b\u003e having fluorine. There is slight decrease in the potency of scaffolds \u003cb\u003e12\u003c/b\u003e (6.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M) and \u003cb\u003e13\u003c/b\u003e (4.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M) observed due to less electronegative nature of chlorine. The \u003cem\u003epara\u003c/em\u003e substituted scaffold is more than \u003cem\u003emeta\u003c/em\u003e substituted scaffold para\u0026thinsp;\u0026gt;\u0026thinsp;meta. The fluorine and chlorine scaffolds (\u003cb\u003e1\u003c/b\u003e, \u003cb\u003e2\u003c/b\u003e, \u003cb\u003e3, 12\u003c/b\u003e and \u003cb\u003e13\u003c/b\u003e) compare with bromine scaffolds (\u003cb\u003e4, 5\u003c/b\u003e and \u003cb\u003e6\u003c/b\u003e) having IC\u003csub\u003e50\u003c/sub\u003e values 10.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M, 13.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M and 9.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M respectively. They showed weak activity. Greater decline was observed in inhibitory potential of compounds having bromine because of less electronegative and big size of bromo \u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. The \u003cem\u003epara\u003c/em\u003e bromo substituted scaffold \u003cb\u003e6\u003c/b\u003e (IC\u003csub\u003e50\u003c/sub\u003e value\u0026thinsp;=\u0026thinsp;9.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u003cem\u003e\u0026micro;\u003c/em\u003eM) is more active than the \u003cem\u003eortho and meta\u003c/em\u003e bromo substituted scaffolds \u003cb\u003e4\u003c/b\u003e and \u003cb\u003e5\u003c/b\u003e (IC\u003csub\u003e50\u003c/sub\u003e value\u0026thinsp;=\u0026thinsp;10.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M, 13.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M) para\u0026thinsp;\u0026gt;\u0026thinsp;ortho\u0026thinsp;\u0026gt;\u0026thinsp;meta.\u003c/p\u003e \u003cp\u003eThe scaffolds having methoxy group as a substituent are \u003cb\u003e7\u003c/b\u003e, \u003cb\u003e8\u003c/b\u003e and \u003cb\u003e9.\u003c/b\u003e The IC\u003csub\u003e50\u003c/sub\u003e values of these scaffolds are 7.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M, 9.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M and 5.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M respectively. The substituted methoxy scaffold follow activity pattern para\u0026thinsp;\u0026gt;\u0026thinsp;ortho\u0026thinsp;\u0026gt;\u0026thinsp;meta. If we compare the methyl substituted compound \u003cb\u003e10\u003c/b\u003e with methoxy scaffolds (\u003cb\u003e7\u0026ndash;9\u003c/b\u003e) we found that that is less active than methoxy compounds this may be due to very less election withdrawing power of methyl\u003c/p\u003e \u003cp\u003eIf nitro substituted scaffolds \u003cb\u003e14\u003c/b\u003e (15.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50 \u0026micro;M) and \u003cb\u003e15\u003c/b\u003e (12.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50\u0026micro;M) are compared with chloro substituted scaffolds (\u003cb\u003e12\u003c/b\u003e and \u003cb\u003e13\u003c/b\u003e). The scaffolds (\u003cb\u003e14\u003c/b\u003e and \u003cb\u003e15\u003c/b\u003e) become much less potent even than the standard is only due to nature of nitro group which not forming good interaction enzyme.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eα-Glucosidase Inhibitory Potential\u003c/h2\u003e \u003cp\u003eAll the synthesized scaffolds (\u003cb\u003e1\u0026ndash;15\u003c/b\u003e) showed good to moderate α-glucosidase inhibitory potential with IC\u003csub\u003e50\u003c/sub\u003e values between 3.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M to 16.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60 \u0026micro;M when compared with standard acarbose having IC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;9.80.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) \u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. The difference between the inhibitory potential of the synthesized scaffolds was studied by structure activity relationship, which depend upon the numbers, nature and positions of substituent/s on the phenyl ring \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe scaffolds (\u003cb\u003e1\u003c/b\u003e, \u003cb\u003e2\u003c/b\u003e and \u003cb\u003e3\u003c/b\u003e) in this series are fluorinated thiazole-based scaffolds having IC\u003csub\u003e50\u003c/sub\u003e values 3.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M, 4.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M and 5.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M respectively. If we compared these three scaffolds with standard, then all these three scaffolds are more potent than standard. The high electronegativity and small size of fluorine is responsible for their more potency. The \u003cem\u003eortho\u003c/em\u003e (3.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M) substituted scaffold is more potent than the \u003cem\u003emeta\u003c/em\u003e (4.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M) and \u003cem\u003epara\u003c/em\u003e (5.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M) substituted scaffolds. The scaffolds (\u003cb\u003e12\u003c/b\u003e and \u003cb\u003e13)\u003c/b\u003e are chlorinated scaffolds of this series having IC\u003csub\u003e50\u003c/sub\u003e values 7.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M and 4.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M respectively. If both chlorinated scaffolds compared with one another, then the \u003cem\u003epara\u003c/em\u003e substituted scaffold is more potent than \u003cem\u003emeta\u003c/em\u003e substituted scaffold (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Although both scaffolds are more potent than standard. If we compare with fluorinated there is slight decrease in activity this may be due to the decrease in electronegativity.\u003c/p\u003e \u003cp\u003eComparing scaffolds (\u003cb\u003e4\u003c/b\u003e, \u003cb\u003e5\u003c/b\u003e and \u003cb\u003e6\u003c/b\u003e) having IC\u003csub\u003e50\u003c/sub\u003e values 11.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M, 13.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40 \u0026micro;M and 9.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M respectively with fluorinated scaffolds (\u003cb\u003e1\u003c/b\u003e, \u003cb\u003e2\u003c/b\u003e and \u003cb\u003e3\u003c/b\u003e) and scaffolds (\u003cb\u003e12\u003c/b\u003e and \u003cb\u003e13)\u003c/b\u003e having chlrine. There is large decrease in their activity take place. The \u003cem\u003epara\u003c/em\u003e bromo substituted scaffolds (9.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M) is more potent even than standard but the \u003cem\u003eortho\u003c/em\u003e and \u003cem\u003emeta\u003c/em\u003e bromo substituted scaffolds less potent than standard\u003c/p\u003e \u003cp\u003eIf the bromine is replaced by methoxy group as in scaffolds (\u003cb\u003e7\u003c/b\u003e, \u003cb\u003e8\u003c/b\u003e and \u003cb\u003e9\u003c/b\u003e) having IC\u003csub\u003e50\u003c/sub\u003e values7.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M, 9.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 \u0026micro;M and 5.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M respectively. All these three scaffolds having less IC\u003csub\u003e50\u003c/sub\u003e values than standard (9.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M). Among the methoxy substituted scaffolds, the \u003cem\u003epara\u003c/em\u003e substituted scaffold is more potent than the \u003cem\u003eortho\u003c/em\u003e and \u003cem\u003emeta\u003c/em\u003e substituted scaffolds. If we compare the potency of \u003cem\u003epara\u003c/em\u003e methyl substituted scaffold (\u003cb\u003e10\u003c/b\u003e) the activity decreases this may be due to the low electronegativity of methyl as compared to methoxy\u003c/p\u003e \u003cp\u003eIf chlorine in the above scaffolds is replaced by nitro group as in scaffolds (\u003cb\u003e14\u003c/b\u003e and \u003cb\u003e15\u003c/b\u003e) having IC\u003csub\u003e50\u003c/sub\u003e values 16.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60 \u0026micro;M and 12.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60 \u0026micro;M respectively. Due to nature of nitro group attached at scaffolds (\u003cb\u003e14\u003c/b\u003e and \u003cb\u003e15\u003c/b\u003e), there is huge decease in their potency occur even though both scaffolds (\u003cb\u003e14\u003c/b\u003e and \u003cb\u003e15\u003c/b\u003e) become less potent than standard.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMolecular docking studies\u003c/h3\u003e\n\u003cp\u003eTo rationalize the structure-activity relationship (SAR) of the most active compounds, molecular docking studies were performed against the α-amylase active site to understand why different substituents and their positions on the phenyl ring led to varied inhibitory potencies. Prior to the docking simulation, a validation step was carried out on the α-amylase crystal structure (PDB ID: 4W93) to ensure the reliability and accuracy of the docking protocol for this study. The native co-crystallized ligand, Montbretin A, was extracted from the active site and then re-docked using the same parameters and grid box settings that were established for the synthesized compounds. The accuracy of the procedure was confirmed by calculating the Root Mean Square Deviation (RMSD) between the top-ranked predicted binding pose of Montbretin A and its original experimental pose within the crystal structure. The resulting low RMSD value of less than 2.0 \u0026Aring; (1.7 \u0026Aring;) indicated that the chosen docking method successfully reproduced the native binding mode, thereby validating the protocol suitability for predicting the binding interactions of the synthesized thiazole inhibitors.\u003c/p\u003e \u003cp\u003eThe most potent inhibitors (\u003cb\u003e1\u003c/b\u003e, \u003cb\u003e2\u003c/b\u003e, and \u003cb\u003e3\u003c/b\u003e) are all fluorinated, yet they exhibit distinct binding modes that appear to explain their rank order of activity (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Compound \u003cb\u003e1\u003c/b\u003e (\u003cem\u003eortho\u003c/em\u003e-fluoro), the most active analog (IC₅₀ = 3.10 \u0026micro;M), binds primarily through extensive hydrophobic interactions, with its bromobenzene and thiazole moieties establishing multiple alkyl and π-alkyl contacts with key residues like Ile235, Tyr151, and Ala307. In contrast, compound 3 (\u003cem\u003epara\u003c/em\u003e-fluoro), another highly potent analog (IC₅₀ = 4.20 \u0026micro;M), utilizes a different strategy dominated by a network of conventional hydrogen bonds with Tyr151, Thr163, and Ile148, alongside electrostatic interactions. Compound \u003cb\u003e2\u003c/b\u003e (\u003cem\u003emeta\u003c/em\u003e-fluoro) also relies on hydrophobic contacts like compound \u003cb\u003e1\u003c/b\u003e but is distinguished by a unique \u003cb\u003eπ-\u003c/b\u003esulfur interaction between its thiourea linkage and the residue Trp58. The docking results suggest that the superior activity of the fluorinated analogs is due to their ability to effectively engage the enzyme through either extensive hydrophobic contacts or a strong hydrogen-bonding network, with the specific position of the fluorine atom dictating which of these binding modes is favored and correlating with their inhibitory potentials.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eAs for less potent analogs such as compounds \u003cb\u003e6\u003c/b\u003e and \u003cb\u003e9\u003c/b\u003e, a more detailed docking analysis of these compounds clarifies their moderate potency in comparison to the leading fluorinated analogs (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). It observed that the \u003cem\u003epara\u003c/em\u003e-bromo substituted compound \u003cb\u003e6\u003c/b\u003e (IC₅₀ = 9.30 \u0026micro;M) and the \u003cem\u003epara\u003c/em\u003e-methoxy substituted compound \u003cb\u003e9\u003c/b\u003e (IC₅₀ = 5.60 \u0026micro;M) could possibly form hydrogen bonds with residues like His305 and Asp300, respectively, in addition to hydrophobic interactions. Compound \u003cb\u003e6\u003c/b\u003e, with a \u003cem\u003epara\u003c/em\u003e-bromo substituent, binds itself firmly through various interactions. It forms two crucial hydrogen bonds between its carbonyl oxygen and the His305 sidechain at a distance of 2.09\u0026Aring;, and another between its thiourea group and Asp300 at the distance of 2.99\u0026Aring;. The compound is further stabilized by electrostatic forces, including a π-cation interaction with His201 (3.02\u0026Aring;) and a π-anion interaction with Asp356 (4.15\u0026Aring;), alongside extensive hydrophobic contacts. Notably, its \u003cem\u003epara\u003c/em\u003e-bromo substituent actively participates in binding through close alkyl interactions with Lys200 (3.91\u0026Aring;) and Ile235 (4.05\u0026Aring;). In a similar fashion, compound \u003cb\u003e9\u003c/b\u003e, the most potent methoxy-substituted analog (IC₅₀ = 5.60 \u0026micro;M), formed a conventional hydrogen bond between its carbonyl oxygen and Asp300 at 2.40\u0026Aring;. The analysis highlights a stabilizing halogen bond between the bromine on the compound main ring with the backbone oxygen from Ile235 at 3.36\u0026Aring;. Its fit is reinforced by numerous hydrophobic contacts, particularly the π-alkyl interactions between its \u003cem\u003epara\u003c/em\u003e-methoxy group and the sidechains of Trp59 (at 3.84\u0026Aring; and 4.81\u0026Aring;). While both compounds \u003cb\u003e6\u003c/b\u003e and \u003cb\u003e9\u003c/b\u003e form multiple significant bonds, the specific nature and geometry of these interactions result in good inhibitory activity. This detailed analysis clarifies that their binding modes, while effective, are less optimal than those of the more potent fluorinated analogs, explaining their position in the overall\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eAll solvents and chemicals used in synthesis purchased from Sigma Aldrich and used without further purification. Analytical grade solvents used for reaction as well as washing. NMR spectra recorded on JEOL spectrometer at 500 MHz and 125 MHz as internal standard TMS used and DMSO-d\u003csub\u003e6\u003c/sub\u003e used as solvent. For HR-EIMS high-resolution mass spectra (electron impact, 60 eV) on a Finnigan MAT-311 A instrument (Germany) were used. For visualization of chromatogram, a UV lamp (Schimazdu, Germany) of wavelength 254/365 used.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eGeneral procedure for the synthesis of thiazole-based scaffolds\u003c/h2\u003e \u003cp\u003eThiazole-thiourea analogs synthesized by treating 2-(2-bromophenyl) thiazole-5-carbohydrazide (1 mmol) with numerous aryl isothiocyanates (1 mmol) in the chloroform (10 mL). The reaction mixture kept on stirring for 3\u0026ndash;4 h. The reaction completion was carefully monitored using thin layer chromatography (TLC). After accomplishment of reaction the solvent evaporated by vacuum rotatory evaporator to afford crude products \u003cb\u003e1\u0026ndash;15\u003c/b\u003e. The crude products were recrystallized in methanol to afford 2-(2-bromophenyl) thiazole-5-carbohydrazide-based thiourea (\u003cb\u003e1\u0026ndash;15\u003c/b\u003e). All synthesized compounds fully characterized by using different spectroscopic methods, \u003csup\u003e1\u003c/sup\u003eHNMR, \u003csup\u003e13\u003c/sup\u003eCNMR, HREI MS.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eCharacterization of thiazole-based scaffolds\u003c/h2\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(2-fluorophenyl)hydrazine-1-carbothioamide (\u003cb\u003e1\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.74 (s, 1H, NH), 9.97 (s, 1H, NH), 9.90 (s, 1H, NH), 8.51 (s, 1H, Ar), 8.45 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;1.8 Hz, 1H, Art), 8.05 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.5 Hz, 1H, Ar), 7.75 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.7 Hz, 1H, Ar), 7.51(t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;6.5 Hz, 2H, Ar), 7.32 (m, 2H, Ar), 6.98 (t, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;7.8 Hz, 1H, Ar). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 213.3, 180.7, 165.1 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;153, C-F), 148.8, 141.0, 141.0, 134.4, 133.3, 131.4, 129.6, 128.6, 126.0, 125.6, 123.2, 122.6, 119.5, 113.8; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBrFN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 449.9620; found: 449.9592.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(3-fluorophenyl)hydrazine-1-carbothioamide (\u003cb\u003e2\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.9 (s, 1H, Ar), 10.05 (s, 1H, NH), 9.85 (s, 1H, NH), 8.65 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5Hz, 1H, Ar), 8.25(d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;1.5Hz, 1H, Ar), 7.38 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;4.5 Hz, 1H, Ar), 7.22(d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;2.0 Hz, 1H, Ar), 7.13(s, 1H, Ar), 7.08 (s, 1H, Ar), 7.02 (m, 2H, Ar), 6.88 (s, 1H, Ar).\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 187.4, 182.1, 166.2 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;133, C-F), 160.1, 149.0, 134.4, 133.3, 131.4, 130.5, 128.7, 128.0, 125.9, 125.6, 124.0, 122.6, 115.6, 115.5; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBrFN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 449.9620; found: 449.9599.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(4-fluorophenyl)hydrazine-1-carbothioamide (\u003cb\u003e3\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHMNR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.81 (s, 1H, NH), 10.17 (s, 1H, NH), 9.85 (s, 1H, NH), 8.53 (s, 1H, Ar), 8.49 (m, 2H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.0, 2.0 Hz, 2H, Ar), 8.01 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 2.5 Hz, 1H, Ar), 7.52 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 2.5 Hz, 1H, Ar), 7.63 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.8, 1H, Ar), 7.52 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 1H, Ar). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); δ 180.9, 171.0, 165.2 (d, \u003cem\u003eJ\u003c/em\u003e\u0026thinsp;=\u0026thinsp;158, C-F), 160.1, 155.7, 148.7, 147.1, 140.5, 134.3, 133.3, 131.4, 129.0, 128.6, 126.1, 125.6, 122.6, 119.5; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBrFN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 449.9620; found: 449.9582.\u003c/p\u003e \u003cp\u003e \u003cem\u003eN\u003c/em\u003e-(2-Bromophenyl)-2-(2-(2-Bromophenyl)thiazole-5-carbonyl)hydrazine-1-carbothioamide (\u003cb\u003e4\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ δ\u003c/em\u003e 10.82 (s, 1H, NH), 10.53 (s, 1H, NH), 10.24 (s, 1H, NH), 8.52 (s, 1H, Ar), 8.45 (s, 1H, Ar), 8.22 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.6, 2.0 Hz, 2H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.6, 2.1 Hz, 1H,Ar), 7.94 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.7 Hz, 2H,Ar), 7.75 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6 Hz, 1H,Ar), 7.49 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 1H, Ar). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e): \u003cem\u003eδ\u003c/em\u003e 180.6, 178.4, 148.8, 146.3, 143.2, 134.3, 133.3, 132.5, 131.4, 130.2, 128.6, 126.2, 125.6, 124.7, 123.5, 122.6, 120.1; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBr\u003csub\u003e2\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 509.8819; found: 509.8766.\u003c/p\u003e \u003cp\u003e \u003cem\u003eN\u003c/em\u003e-(3-Bromophenyl)-2-(2-(2-Bromophenyl)thiazole-5-carbonyl)hydrazine-1-carbothioamide (\u003cb\u003e5\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.75 (s, 1H, NH), 9.99 (s, 1H, NH), 9.95 (s, 1H, NH), 8.51 (s, 1H, Ar), 8.46 (s, 1H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.0 Hz, 1H, Ar), 7.75 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.6, 2.6 Hz, 2H, Ar), 7.52\u0026ndash;7.47 (m, 2H, Ar), 7.37 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.7, 1H, Ar), 7.21 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.8, 2.3 Hz, 1H, Ar). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); δ180.7, 165.1, 163.6, 151.1, 148.9, 140.7, 134.3, 132.0, 133.3, 131.4, 129.3, 128.6, 126.0, 125.6, 125.6, 122.6, 120.1; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBr\u003csub\u003e2\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 509.8819; found: 509.8773.\u003c/p\u003e \u003cp\u003e \u003cem\u003eN\u003c/em\u003e-(4-Bromophenyl)-2-(2-(2-Bromophenyl)thiazole-5-carbonyl)hydrazine-1-carbothioamide (\u003cb\u003e6\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); δ 10.72 (s, 1H, Ar), 9.91 (s, 1H, NH), 9.83 (s, 1H, NH), 8.49 (s, 1H Ar), 8.45 (s, 1H Ar), 8.39 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;3.0 Hz, 1H, Ar), 8.32 (s, 1H Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.7, 2.0 Hz, 1H, Ar), 8.02 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.0 Hz, 1H, Ar), 7.75 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.6, 1H, Ar), 7.52\u0026ndash;7.44 (m, 1H, Ar), 7.17 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.3, 1H, Ar). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); δ 180.8, 165.2, 159.6, 149.6, 139.8, 134.5, 133.3, 133.2, 131.4, 131.3, 128.6, 125.9, 125.6, 125.6, 124.0, 122.6, 122.5; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBr\u003csub\u003e2\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 509.8819; found: 509.8781.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(2-methoxyphenyl)hydrazine-1-carbothioamide (\u003cb\u003e7\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.68 (s, 1H, NH), 9.91 (s, 1H, NH), 9.71 (s, 1H, NH), 8.49 (s, 1H Ar), 8.46 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;3.2 Hz, 2H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.3 Hz, 1H, Ar), 7.75 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.7, 2.5 Hz, 1H, Ar), 7.52 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 1H, Ar), 7.31(d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.0 Hz, 2H, Ar), 6.90 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.5 Hz, 2H, Ar), 3.74 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 180.6, 165.0, 161.3, 156.6, 149.1, 140.2, 134.4, 133.3, 132.0, 131.4, 131.3, 128.6, 125.8, 125.6, 122.6, 120.1, 113.1, 55.4; HREI-MS: m/z calculated for C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e15\u003c/sub\u003eBrN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003eS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 461.9820; found: 461.9761.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(3-methoxyphenyl)hydrazine-1-carbothioamide (\u003cb\u003e8\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.70 (s, 1H, NH), 9.84 (s, 1H, NH), 9.71 (s, 1H, NH), 8.50 (s, 1H, Ar), 8.45 (s, 1H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.2 Hz, 1H, Ar), 7.74 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 2.5 Hz, 1H, Ar), 7.52 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 1H, Ar), 7.24 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.7, 2H, Ar), 7.09 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.7, 2.3 Hz, 1H, Ar), 6.74 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.9, 2.2 Hz, 1H, Ar), 3.75 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); δ 181.4, 165.1, 158.8, 149.1, 144.2, 140.3, 134.4, 133.3, 131.4, 130.5, 130.2, 129.8, 128.6, 128.4, 125.9, 125.6, 122.6, 55.0; HREI-MS: m/z calculated for C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e15\u003c/sub\u003eBrN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003eS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 461.9820; found: 461.9777.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(4-methoxyphenyl)hydrazine-1-carbothioamide (\u003cb\u003e9\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.89 (s, 1H, NH), 10.05 (s, 1H, NH), 9.85 (s, 1H, Ar), 9.24 (s, 1H, Ar), 8.52 (s, 1H, Ar), 8.45 (s, 1H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.2 Hz, 1H, Ar), 7.75 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.7, 2.4 Hz, 1H, Ar), 7.51 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6 Hz, 1H, Ar), 7.34 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.2 Hz, 1H, Ar), 7.18\u0026ndash;7.15 (m, 1H, Ar), 6.93 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.4, 1H, Ar), 3.73 (s, 3H, OCH\u003csub\u003e3\u003c/sub\u003e). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); δ 181.0, 165.1, 155.5, 148.7, 134.4, 133.4, 131.4, 129.1, 128.7, 126.1, 125.6, 125.5, 122.6, 120.8, 119.7, 112.3, 111.4, 55.6; HREI-MS: m/z calculated for C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e15\u003c/sub\u003eBrN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003eS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 461.9820; found: 461.9752.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(p-tolyl)hydrazine-1-carbothioamide (\u003cb\u003e10\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.69 (s, 1H, NH), 9.91 (s, 1H, NH), 9.75 (s, 1H, NH), 8.49 (s, 1H Ar), 8.40 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;2.0 Hz, 1H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.3 Hz, 1H, Ar), 8.01 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;2.3 Hz, 1H, Ar), 7.74\u0026ndash;7.71 (m, 1H, Ar), 7.52\u0026ndash;7.47 (m, 2H, Ar), 7.13 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.9 Hz, 1H, Ar), 2.28 (s, 3H, CH\u003csub\u003e3\u003c/sub\u003e).\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 181.3, 165.2, 159.6, 149.6, 134.5, 134.4, 133.3, 133.2, 131.4, 131.3, 128.7, 125.8, 125.6, 125.6, 124.0, 122.6, 122.5, 20.5; HREI-MS: m/z calculated for C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e15\u003c/sub\u003eBrN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 445.9871; found: 445.9844.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-phenylhydrazine-1-carbothioamide (\u003cb\u003e11\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.74 (s, 1H, NH), 9.91 (s, 1H, NH), 9.87 (s, 1H, NH), 8.50 (s, 1H Ar), 8.45 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;2.0 Hz, 1H, Ar), 8.39 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.5 Hz, 1H, Ar), 8.32 (m, 1H, Ar), 8.06 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;2.2 Hz, 1H, Ar), 8.02 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.0 Hz, 1H, Ar), 7.73 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.5, 2.0 Hz, 1H, Ar), 7.49 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 2H, Ar). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 180.3, 165.2, 159.6, 149.6, 134.5, 133.2, 132.7, 131.3, 130.2, 128.6, 128.5, 128.5, 126.4, 125.6, 125.6, 124.0, 122.5; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e13\u003c/sub\u003eBrN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 431.9714; found: 431.9667.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(3-chlorophenyl)hydrazine-1-carbothioamide (\u003cb\u003e12\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.78 (s, 1H, NH), 9.93 (s, 1H, NH), 9.67 (s, 1H, NH), 8.51 (s, 1H Ar), 8.47 (s,1H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.2 Hz, 1H, Ar), 7.74 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 2.5 Hz, 1H, Ar), 7.65 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.0 Hz, 1H, Ar), 7.52 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5 Hz, 1H, Ar), 7.42\u0026ndash;7.37 (m, 2H, Ar), 7.21(t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.6 Hz, 1H, Ar).\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 181.0, 165.0, 160.1, 149.0, 134.4, 133.3, 132.3, 132.0, 130.2, 129.8, 131.4, 128.7, 127.6, 127.6, 126.0, 125.6, 122.6; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBrClN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 465.9324; found: 465.9301.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(4-chlorophenyl)hydrazine-1-carbothioamide (\u003cb\u003e13\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.74 (s, 1H, NH), 9.99 (s, 1H, NH), 9.90 (s, 1H, NH), 8.51 (s, 1H Ar), 8.46 (s,1H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.1 Hz, 1H, Ar), 7.75 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.7, 2.4 Hz, 2H, Ar), 7.56 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.9 Hz, 1H, Ar), 7.51 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5 Hz, 1H, Ar), 7.35 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.9, 1.3 Hz, 1H, Ar), 7.29 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6 Hz, 1H, Ar). \u003csup\u003e13\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 170.8, 169.4, 165.2, 148.9, 147.1, 140.9, 140.5, 140.5, 134.4, 133.4, 132.2, 131.4, 128.6, 128.6, 125.7, 125.6, 122.6; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBrClN\u003csub\u003e4\u003c/sub\u003eOS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 465.9324; found: 465.9295.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(3-nitrophenyl)hydrazine-1-carbothioamide (\u003cb\u003e14\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.73(s, 1H, NH), 9.92 (s, 1H, NH), 9.88 (s, 1H, NH), 8.57 (s, 1H, Ar), 8.50 (s, 1H, Ar), 8.32 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;3.3 Hz, 1H, Ar), 8.06 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.2 Hz, 1H, Ar), 7.74 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6, 2.5 Hz, 2H, Ar), 7.52 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.6 Hz, 1H, Ar), 7.39 (d \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;8.2, 2.7 Hz, 2H, Ar).\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 181.7, 165.1, 149.0, 144.2, 143.5, 138.2, 134.3, 133.3, 131.4, 129.5., 129.2, 128.6, 127.7, 125.9, 125.6, 122.6, 120.3; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBrN\u003csub\u003e5\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003eS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 476.9565; found: 476.9546.\u003c/p\u003e \u003cp\u003e2-(2-(2-Bromophenyl)thiazole-5-carbonyl)-\u003cem\u003eN\u003c/em\u003e-(4-nitrophenyl)hydrazine-1-carbothioamide (\u003cb\u003e15\u003c/b\u003e)\u003c/p\u003e \u003cp\u003e \u003csup\u003e1\u003c/sup\u003eHNMR (500 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 10.71 (s, 1H, NH), 9.91 (s, 1H, NH), 9.82 (s, 1H, NH), 8.50 (s, Ar), 8.46 (d, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;1.5Hz, 1H, Ar), 8.05 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.5, 2.3Hz, 1H, Ar), 7.74 (dd, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;7.4, 2.3 Hz, 1H, Ar), 7.52\u0026ndash;7.46 (m, 2H, Ar), 7.33 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.8 Hz, 2H, Ar), 7.15 (t, \u003cem\u003eJ\u0026thinsp;=\u003c/em\u003e\u0026thinsp;6.0 Hz, 1H, Ar).\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003eCNMR (125 MHz, DMSO-d\u003csub\u003e6\u003c/sub\u003e); \u003cem\u003eδ\u003c/em\u003e 180.2, 165.1, 149.1, 148.7, 148.7, 139.2, 134.4, 133.4, 131.4, 128.6, 127.8, 125.9, 125.6, 124.4, 123.1, 123.1, 122.6; HREI-MS: m/z calculated for C\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eBrN\u003csub\u003e5\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003eS\u003csub\u003e2\u003c/sub\u003e [M]\u003csup\u003e+\u003c/sup\u003e 476.9565; found: 476.9533.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eBiological activity\u003c/h2\u003e \u003cp\u003eα-Amlyase and α-glucosidase inhibition assay inhibitory activity Enzyme and reagents purchased from sigma Aldrich, we used the same protocol reported in our previous paper \u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e,\u003cspan additionalcitationids=\"CR26\" citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e for detail kindly see supplementary data.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eMolecular Docking\u003c/h2\u003e \u003cp\u003eIn this study AutoDock 4.2 \u003csup\u003e28\u003c/sup\u003e was utilized to dock the small molecules into the active site of the protein. A homology model of α-glucosidase that was constructed based on the crystal structure of isomaltase from \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e (PDB ID: 3A4A) using SwissModel \u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. While for α-amylase enzyme, the crystal structure (PDB ID: 4W93) was downloaded from protein databank website. The structure of α-amylase enzyme was optimized by removing water molecules, adding hydrogen atoms, adding charge, and repairing end residues. For α-glucosidase enzyme, the grid map was generated based on 0.375 \u0026Aring; spacing between grid points, and the center of the grid box was placed at coordinate x\u0026thinsp;=\u0026thinsp;14.885107, y = -11.366557, and z\u0026thinsp;=\u0026thinsp;18.413239. The dimensions of the active site box were set at 60 \u0026times; 60 \u0026times; 60 \u0026Aring;. While for α-amylase enzyme, the grid map was generated using 0.375 \u0026Aring; spacing between grid points, and the center of the grid box was placed at coordinate x = -9.632488, y\u0026thinsp;=\u0026thinsp;4.340907, and z = -23.107256. Each docked system was performed with 150 runs using Lamarckian genetic algorithm function. Protein‒ligand interactions were visualized and analyzed using Discovery Studio Visualizer 3.5.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn this work fifteen thiazole-based scaffolds (\u003cb\u003e1\u003c/b\u003e\u0026ndash;\u003cb\u003e15\u003c/b\u003e) have been synthesized and characterized using various spectroscopic techniques including \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003eHNMR, \u003csup\u003e13\u003c/sup\u003eCNMR, HREI MS. Then all these synthesized scaffolds were screened for their alpha amylase and alpha glucosidase inhibitory activity. All synthesized thiazole-based scaffolds displayed wide range of inhibition for both targeted enzymes. The IC\u003csub\u003e50\u003c/sub\u003e values for alpha amylase and alpha glucosidase are 3.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M to 15.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50 \u0026micro;M and 3.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M to 15.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50 \u0026micro;M respectively. The scaffolds (\u003cb\u003e1\u003c/b\u003e, \u003cb\u003e2\u003c/b\u003e and \u003cb\u003e3\u003c/b\u003e) have been found most potent in the synthesized series of thiazole-based scaffolds. These are fluorinated thiazole-based scaffolds in which the fluorine might be responsible for their potency. The binding interactions between most potent scaffold and targeted enzymes have been studied through molecular docking, which confirmed the binding interaction of compounds with enzymes\u0026rsquo; active site.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eFunding:\u003c/h2\u003e \u003cp\u003eThis research did not receive funding.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eM. T and F. R (Synthesis); M. A and S. A. A. S. (Characterization of compounds); N. U. and F. S. A (Purification of compounds); A. S. and K. M. K. (Bioassay); S. I. (Molecular docking); M. T, M. A. and K. M. K. Writing and editing manuscript\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eAuthors would like to thank Imam Abdulrahman Bin Faisal University for providing excellent lab facilities.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe datasets generated and/or analyzed during the current study are not publicly available because they are private, but are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eWondafrash, D. Z. et al. Potential effect of hydroxychloroquine in diabetes mellitus: a systematic review on preclinical and clinical trial studies. \u003cem\u003eJ. Diabetes Res.\u003c/em\u003e 5214751 (2020). (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEvans-Molina, C. et al. 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Synthesis, in vitro evaluation and molecular docking studies of hybrid 4-quinolinyl bearing 1, 3, 4-thiadiazole-2-amine as a new inhibitor of α-amylase and α-glucosidase. \u003cem\u003eJ. Mol. Struct.\u003c/em\u003e \u003cb\u003e1282\u003c/b\u003e, 135173 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMukhliss, L. et al. Synthesis and novel structural hybrid analogs of oxindole derivatives bearing piperidine ring, their antidiabetics II activity and molecular docking study. \u003cem\u003eJ. Mol. Struct.\u003c/em\u003e \u003cb\u003e1332\u003c/b\u003e, 141666 (2025).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaleem, F. et al. Bioevaluation of synthetic pyridones as dual inhibitors of α-amylase and α‐glucosidase enzymes and potential antioxidants. \u003cem\u003eArch. Pharm.\u003c/em\u003e \u003cb\u003e356\u003c/b\u003e, 2200400 (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJavid, M. T. et al. Synthesis, in vitro α-glucosidase inhibitory potential and molecular docking study of thiadiazole analogs. \u003cem\u003eBioorg. Chem.\u003c/em\u003e \u003cb\u003e78\u003c/b\u003e, 201\u0026ndash;209 (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRahim, F. et al. Triazinoindole analogs as potent inhibitors of α-glucosidase: Synthesis, biological evaluation and molecular docking studies. \u003cem\u003eBioorg. Chem.\u003c/em\u003e \u003cb\u003e58\u003c/b\u003e, 81\u0026ndash;87 (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMorris, G. M. et al. AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility. \u003cem\u003eJCoCh\u003c/em\u003e \u003cb\u003e30\u003c/b\u003e, 2785\u0026ndash;2791 (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWaterhouse, A. et al. SWISS-MODEL: homology modelling of protein structures and complexes. \u003cem\u003eNAR\u003c/em\u003e 46, W296-W303 (2018).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Table 1","content":"\u003cp\u003eTable 1 is available in the Supplementary Files section.\u003c/p\u003e"},{"header":"Schemes","content":"\u003cp\u003eScheme 1 is available in the Supplementary Files section\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Thiazole, antidiabetic activity, SAR, molecular docking","lastPublishedDoi":"10.21203/rs.3.rs-8601560/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8601560/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn this research work fifteen thiazole-based scaffolds (\u003cb\u003e1\u0026ndash;15\u003c/b\u003e) were synthesized and evaluated for the α-amylase and α-glucosidase inhibitory potential. In this series all the synthesized scaffolds showed good to moderate α-amylase and α-glucosidase inhibition and their IC\u003csub\u003e50\u003c/sub\u003e value ranging between 3.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M to 15.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50 \u0026micro;M for α-amylase and IC\u003csub\u003e50\u003c/sub\u003e value from 3.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M to 16.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60 \u0026micro;M for α-glucosidase when compared with the standard acarbose having IC\u003csub\u003e50\u003c/sub\u003e value 10.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M (for α-amylase) and 9.80.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M (for α-glucosidase). Among the synthesized scaffolds, nine analogs (\u003cb\u003e1\u003c/b\u003e\u0026ndash;\u003cb\u003e3\u003c/b\u003e, \u003cb\u003e6\u003c/b\u003e\u0026ndash;\u003cb\u003e9\u003c/b\u003e, \u003cb\u003e12\u003c/b\u003e and \u003cb\u003e13\u003c/b\u003e) were found more active against both enzymes. The scaffolds \u003cb\u003e1\u003c/b\u003e IC\u003csub\u003e50\u003c/sub\u003e value (3.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M, 3.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M), \u003cb\u003e2\u003c/b\u003e IC\u003csub\u003e50\u003c/sub\u003e value (4.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M, 4.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M) and \u003cb\u003e3\u003c/b\u003e IC\u003csub\u003e50\u003c/sub\u003e value (4.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 \u0026micro;M, 5.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 \u0026micro;M) were found most potent among this series against both α-amylase and α-glucosidase enzymes respectively. These scaffolds having fluorine at \u003cem\u003eortho\u003c/em\u003e, \u003cem\u003emeta\u003c/em\u003e and \u003cem\u003epara\u003c/em\u003e position of the phenyl ring respectively. Furthermore, the structures of all the synthesized analogs were confirmed by using \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003eH-NMR, \u003csup\u003e13\u003c/sup\u003eC-NMR spectroscopy and HR-MS. To study the binding mode of interaction between active site of the targeted enzyme and most potent scaffold, molecular docking study were conducted.\u003c/p\u003e","manuscriptTitle":"Identification thiazole-based analogs as potent antidiabetic agents and their molecular docking study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-16 12:50:16","doi":"10.21203/rs.3.rs-8601560/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2026-02-14T18:05:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"28847950360470235516772888356413814542","date":"2026-02-11T18:42:27+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-11T07:34:06+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-01-28T15:24:41+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-01-24T06:36:00+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-24T06:33:40+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2026-01-14T11:54:16+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"dc64c139-ddf1-42db-8030-55c86f21fbcc","owner":[],"postedDate":"February 16th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":62940207,"name":"Biological sciences/Biochemistry"},{"id":62940208,"name":"Biological sciences/Chemical biology"},{"id":62940209,"name":"Physical sciences/Chemistry"},{"id":62940210,"name":"Biological sciences/Drug discovery"}],"tags":[],"updatedAt":"2026-04-07T17:23:33+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-16 12:50:16","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8601560","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8601560","identity":"rs-8601560","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}