System for the preparation of piperazine-linked imidazo[2,1-b]thiazole derivatives as anticancer, antioxidant, and anti-inflammatory agents

Abstract

A system (100) for the preparation of piperazine-linked imidazo[2,1-b]thiazole derivatives as anticancer agents, antioxidants and anti-inflammatory agents, comprising: a) a reaction chamber (102) configured to enable multiple chemical reactions for the synthesis of piperazine-linked imidazo[2,1-b]thiazole derivatives; b) a heating unit (104) operatively connected to the reaction chamber (102) and configured to provide controlled temperature conditions ranging from room temperature to reflux temperature; c) a cooling unit (106) operatively connected to the reaction chamber (102) and configured to maintain cooling temperatures during operation of the reaction chamber (102); d) a stirring unit (108) operatively connected to the reaction chamber (102) and configured to provide continuous mixing of the reaction components; e) a solvent supply system (110) configured to supply reaction solvent to the reaction chamber (102); f) a reagent supply system (112) configured to supply reaction reagents to the reaction chamber (102); g) a purification unit (114) configured to purify the synthesized piperazine-linked imidazo[2,1-b]thiazole derivatives by column chromatography; and h) a sample collection unit (116) configured to collect and store the resulting product after each reaction performed in the reaction chamber, the sample collection unit (116) being connected to a delivery system configured to return the collected sample to the reaction chamber for subsequent reactions.

Description

FIELD OF THE INVENTIONThe present disclosure relates to a system for the preparation of piperazine-bound imidazo[2,1-b]thiazole derivatives as agents for combating cancer, as antioxidants and as anti-inflammatory active ingredients. More particularly, the present invention relates to the synthesis of 6-phenylimidazo[2,1-b]thiazolamide derivatives (11a-1) and to the evaluation of their in vitroanti-cancer activity against the human cancer cell lines MCF-7 (breast), K-562 (leukemia) and COLO-205 (large intestine) as well as their antioxidant and anti-inflammatory activity.BACKGROUND OF THE INVENTIONCancer is the second leading cause of death worldwide. It is anticipated that the mortality rates will exceed 13 million annually up to 2030. This is attributable in particular to an unhealthy lifestyle with poor nutrition, lack of exercise and substance abuse. Existing chemotherapeutic agents often have significant disadvantages such as lack of selectivity, high toxicity and the development of resistances. Also, widely used non-steroidal anti-rheumatizing agents (NSTAR) are associated with severe gastrointestinal and systemic side effects, limiting their long term use. Moreover, oxidative stress caused by an imbalance of reactive oxygen species (ROS) is known to play a critical role in the development of cancer, inflammation and other chronic diseases.Heterocyclic compounds containing nitrogen and sulfur atoms have gained great importance in recent years because of their diverse pharmacological profiles in medical chemistry. Imidazole and thiazole derivatives have shown strong anti-cancer, anti-inflammatory, anti-oxidative and antimicrobial properties. Moreover, incorporation of piperazine moieties into heterocyclic structures has been shown to enhance biological activity and receptor binding by improved structural flexibility and pharmacokinetic profiles.In view of the foregoing discussion, the invention provides a system for synthesizing piperazine-linked imidazo[2,1-b]thiazole derivatives. These hybrid molecules combine pharmacologically-favored frameworks in a single framework with the aim of achieving synergistic effects against cancer and inflammations while providing antioxidant protection.SUMMARY OF THE INVENTIONThe present disclosure relates to a system for the preparation of piperazine-linked imidazo[2,1-b]thiazole derivatives as anti-cancer, anti-oxidation and anti-inflammatory drugs. The system includes special equipment, including a reaction chamber for multiple chemical reactions, temperature control units, mixing mechanisms, solvent and reagent delivery systems, and cleaning components that cooperate to allow efficient and controlled synthesis of these therapeutic compounds.The present disclosure aims to provide a system for the preparation of piperazine-linked imidazo[2,1-b]thiazole derivatives as anti-cancer agents, antioxidants and inflammations. The system comprises: a) a reaction chamber configured to allow for multiple chemical reactions for the synthesis of piperazine-linked imidazo[2,1-b]thiazole derivatives; b) a heating unit operatively connected to the reaction chamber and configured to provide controlled temperature conditions from room temperature to reflux temperature; c) a cooling unit operatively connected to the reaction chamber and configured to maintain cooling temperatures during operation of the reaction chamber; d) a stirring unit operatively connected to the reaction chamber and configured to allow for continuous mixing of the reaction components; e) a solvent delivery system configured to provide reaction solvent to the reaction chamber; f) a reagent delivery system configured to provide reaction reagents to the reaction chamber; g) a purification unit configured to purify the synthesized piperazine-linked imidazo[2,1-b]thiazole derivatives by column chromatography; and h) a sample collection unit configured to collect and store the resultant product after each reaction performed in the reaction chamber, wherein the sample collection unit is connected to a supply system configured to re-supply the collected sample to the reaction chamber for subsequent reactions.An object of the present disclosure is to provide a system for preparing piperazine-linked imidazo[2,1-b]thiazole derivatives as anti-cancer agents, as antioxidants and as anti-inflammatory agents.Another object of the present disclosure is to provide an integrated system for synthesizing piperazine-linked imidazo[2,1-b]thiazole derivatives by a controlled sequential reaction process.Another object of the present disclosure is to provide a system that enables precise control of temperature, mixing and reagent delivery over multiple reaction stages to optimize yield and purity of the final compounds.Another object of the present disclosure is to provide an optimized synthetic route system for the preparation of anti-cancer, anti-oxidation and anti-inflammatory compounds by automated reaction monitoring and purification processes.In order to further clarify the advantages and features of the present disclosure, the invention will be explained in more detail with reference to specific embodiments that are illustrated in the accompanying drawings. These drawings illustrate only typical embodiments of the invention and are therefore not to be considered as limiting the scope thereof. The invention will be described and explained in more detail with reference to the accompanying drawings.BRIEF DESCRIPTION OF THE FIGURESThese and other features, aspects, and advantages of the present disclosure will become more fully understood when the following detailed description is read with reference to the accompanying drawings, in which like characters represent like parts throughout. The following applies here: FIG. 1 illustrates a block diagram of a system for preparing piperazine-linked imidazo[2,1-b]thiazole derivatives as anti-cancer agents, antioxidant, and anti-inflammatory agents according to an embodiment of the present disclosure; FIG. 2A illustrates a scheme for synthesizing (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone and its acetamide derivatives according to an embodiment of the present disclosure; and FIG. 2B illustrates structures of reagents used for the synthesis of (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone and its acetamide derivatives according to an embodiment of the present disclosure.Those skilled in the art will also appreciate that the elements in the drawings are shown for simplicity and are not necessarily to scale. For example, the flowcharts illustrate the method using the key steps to improve understanding of aspects of the present disclosure. In addition, regarding the construction of the apparatus, individual or multiple components of the apparatus may be represented by conventional symbols in the drawings. The drawings may only show the specific details relevant to understanding the embodiments of the present disclosure in order not to obscure the drawings with details readily apparent to those skilled in the art after the present description.DETAILED DESCRIPTION:In order to aid in the understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the drawings and will be described in an comprehensible manner. However, the scope of the invention is not limited thereby. Changes and further modifications of the illustrated system, as well as further applications of the principles of the invention, are possible, as would normally occur to a person skilled in the art.It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the invention and are not intended to be limiting thereof.References throughout this specification to "one aspect," "another aspect," or similar language mean that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, the phrases "in one embodiment," "in another embodiment," and similar phrases in this specification may or may not refer to the same embodiment.The terms "comprises," "comprising," or other variations thereof are intended to cover a non-exclusive inclusion, such that a process or method comprising a list of steps may include not only those steps, but also other steps not expressly listed or inherent in that process or method. Likewise, the phrase "comprises... for" one or more devices, subsystems, elements, structures, or components does not exclude, without further limitations, the existence of other devices, subsystems, elements, structures, components, or additional devices, subsystems, elements, structures, or components.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as understood by one of ordinary skill in the art. The systems, methods, and examples provided herein are for illustrative purposes only and are not to be considered limiting.Embodiments of the present disclosure will be described in detail below with reference to the accompanying drawings.FIG. 1 illustrates a block diagram of a system (100) for preparing piperazine-linked imidazo[2,1-b]thiazole derivatives as anti-cancer agents, antioxidant, and anti-inflammatory agents according to an embodiment of the present disclosure.Referring to FIG. 1, the system (100) comprises: a) a reaction chamber (102) configured to allow multiple chemical reactions for the synthesis of piperazine-linked imidazo[2,1-b]thiazole derivatives; b) a heating unit (104) operatively connected to the reaction chamber (102) and configured to provide controlled temperature conditions ranging from room temperature to reflux temperature; c) a cooling unit (106) operatively connected to the reaction chamber (102) and configured to maintain cooling temperatures during operation of the reaction chamber (102); d) a stirring unit (108) operatively connected to the reaction chamber (102) and configured to provide continuous mixing of the reaction components; e) a solvent delivery system (110) configured to deliver reaction solvent to the reaction chamber (102); f) a reagent supply system (112) configured to supply reaction reagents to the reaction chamber (102); g) a purification unit (114) configured to purify the synthesized piperazine-linked imidazo[2,1-b]thiazole derivatives by column chromatography; and h) a sample collection unit (116) configured to collect and store the resulting product after each reaction performed in the reaction chamber, wherein the sample collection unit (116) is connected to a delivery system configured to deliver the collected sample to the reaction chamber again for subsequent reactions.In one embodiment, the reaction chamber (102) is configured to facilitate the synthesis of ethyl 2-aminothiazole-4-carboxylate (Compound 3), wherein the reaction between ethyl 3-bromo-2-oxopropanoate and thiourea is carried out at 100° C. for 3 hours to produce ethyl 2-aminothiazole-4-carboxylate.In one embodiment, reaction chamber (102) is further configured to facilitate the reaction between ethyl 2-aminothiazole-4-carboxylate (Compound 3) and 2-bromo-1-phenylethanone in the presence of sodium bicarbonate at reflux temperature for 8 hours to synthesize ethyl 6-phenylimidazo[2,1-b]thiazole-3-carboxylate (Compound 5).In one embodiment, the reaction chamber (102) is further configured to allow a reaction for converting ethyl 6-phenylimidazo[2,1-b]thiazole-3-carboxylate (Compound 5) to 6-phenylimidazo[2,1-b]thiazole-3-carboxylic acid (Compound 6) using lithium hydroxide at room temperature for 12 hours, wherein ethyl 6-phenylimidazo[2,1-b]thiazole-3-carboxylate (Compound 5) is hydrolyzed in the presence of THF, methanol, water, and lithium hydroxide.In one embodiment, reaction chamber (102) is further configured to allow a reaction between 6-phenylimidazo[2,1-b]thiazole-3-carboxylic acid (Compound 6) and tert-butylpiperazine-1-carboxylate (Compound 7) in the presence of DMF, HATU, and DIPEA to synthesize tert-butyl 4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazine-1-carboxylate (Compound 8).In one embodiment, reaction chamber (102) is further configured to allow a reaction to convert tert-butyl 4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazine-1-carboxylate (Compound 8) to (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone hydrochloride (Compound 9) using HCl in 1,4-dioxane at room temperature for 12 hours.In one embodiment, system (100) further comprises a secondary reaction chamber (102a) configured to facilitate the synthesis of substituted 2-chloro-N-phenylacetamide compounds [10 (al)] by reaction of trifluoroacetyl chloride with substituted anilines in the presence of DMF and DIPEA.In one embodiment, reaction chamber (102) is also configured to facilitate synthesis of the final target compound by allowing a reaction between (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone hydrochloride (compound 9) and substituted 2-chloro-N-phenylacetamide compounds [10 (al)] at 60°C for 8 hours.In one embodiment, the system (100) further comprises a precipitation unit (118) connected to the reaction chamber (102), a purification unit (114), and a sample collection unit (116) configured to isolate reaction products by adding reaction mixtures to ice-cold water and collecting the resulting precipitates.In one embodiment, the system (100) further comprises a monitoring unit (120) connected to the reaction chamber (102) and configured to track the course of the reaction by thin layer chromatography (TLC) with visualization under UV light, iodine, and ninhydrin.The present invention relates to a specific system for the synthesis of piperazine-linked imidazo[2,1-b]thiazole derivatives having significant pharmacological properties. The frog of the system is a versatile reaction chamber in which several chemical transformations required for the synthetic route can be carried out. This reaction chamber is equipped with an integrated heating unit which can ensure precisely controlled temperature conditions from room temperature to reflux, which are essential for various reactions in the synthetic route. For temperature sensitive reactions requiring cooling, especially during the addition of reagents, the system contains a special cooling unit which keeps the reaction mixture at 0°C thus preventing undesirable side reactions and ensuring selectivity. The system has a sophisticated agitator unit connected to the reaction chamber which provides continuous and uniform mixing of the reaction components, thereby improving reaction efficiency and yield. The automated solvent delivery system delivers various reaction media including ethanol, THF, methanol, DMF and 1,4-dioxane as needed for the various reaction phases, while the reagent delivery system delivers important reagents including thiourea, sodium bicarbonate, lithium hydroxide, HATU, DIPEA and HCl precisely at the proper time and at the proper concentration.FIG. 2A illustrates a scheme for synthesizing (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone and its acetamide derivatives according to an embodiment of the present disclosure.FIG. 2B illustrates structures of reagents used for the synthesis of (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone and its acetamide derivatives according to an embodiment of the present disclosure.Referring to Figs. 2A and 2B, the piperazine-bound imidazo[2,1-b]thiazole derivative synthesizing system comprises an integrated structure having a reaction chamber, a heating unit, a cooling unit, a stirring unit, a solvent supply system, a reagent supply system, a cleaning unit, a sampling unit, a precipitation unit, and a monitoring unit. All components are operatively linked to allow sequential synthesis, isolation and purification of the target compounds. In Figure 2B: a) ethanol, 100°C, RT, overnight, 50-70%; b) EtOH, NaHCO3, reflux, 8 h, 86%; c) THF, methanol, H 2 O, LiOH, RT, 12 h; d) DMF, HATU, DIPEA, RT, 12 h, 81%; e) 1,4-dioxane, HCl, room temperature, 12 h; f) DMF, DIPEA, 60°C, 8 h. In addition, the reaction depicted under the reagent structure shows the synthesis of reagent (f).The reaction chamber allows the reaction between thiourea and ethyl 3-bromo-2-oxopropanoate to synthesize ethyl 2-aminothiazole-4-carboxylate (Compound 3). Thiourea is placed in ethanol and ethyl 3-bromo-2-oxopropanoate is added portionwise over a period of 20 minutes. The heating unit maintains the reaction temperature at 100° C. for three hours while the stirring unit provides continuous mixing. After completion of the reaction, the cooling unit brings the reaction mixture to room temperature. A cream-colored precipitate is formed, which is collected by the sampling unit. The precipitate is washed with 200 ml hexane to give a white solid identified as compound 3.The reaction chamber then allows reaction between compound 3 and 2-bromo-1-phenylethanone in the presence of sodium bicarbonate and ethanol to synthesize ethyl 6-phenylimidazo[2,1-b]thiazole-3-carboxylate (compound 5). Sodium bicarbonate is introduced in a molar ratio of 2:1 relative to compound 3. The heating unit keeps the reaction at reflux temperature for 8 hours with constant stirring. After cooling, ethyl acetate is added to the reaction mixture. The purification unit washes the mixture with water, dries the organic layer over sodium sulfate and concentrates it under vacuum. The crude product is then subjected to column chromatography using silica gel (100-200 mesh) and 50% ethyl acetate in hexane as eluant. The purified compound is dried and collected as compound 5.The reaction chamber also facilitates the hydrolysis of compound 5 to obtain 6-phenylimidazo[2,1-b]thiazole-3-carboxylic acid (compound 6). A stirred solution of compound 5 is prepared in tetrahydrofuran (THF) and MeOH. Lithium hydroxide and water are added to this solution and the mixture is stirred at room temperature for 12 hours. The monitoring unit keeps track of the course of the reaction by thin-layer chromatography (TLC). After completion, the reaction mixture is concentrated under vacuum. The resulting residue is dissolved in 25 ml of water and acidified to pH 2-3 with 2N hydrochloric acid to effect precipitation. The precipitation unit isolates the precipitate, which is filtered, dried under vacuum and triturated with 30 ml hexane to give pure compound 6.The reaction chamber then facilitates coupling of compound 6 with tert-butylpiperazine-1-carboxylate (compound 7) to synthesize tert-butyl 4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazine-1-carboxylate (compound 8). A stirred solution of compound 6 is prepared in dimethylformamide (DMF). HATU and DIPEA are added at 0°C. After stirring at room temperature for 5 minutes, compound 7 is added. The mixture is stirred for 12 hours. The monitor tracks the course of the reaction by TLC using 70% ethyl acetate in hexane and visualization under UV light and iodine. After completion, the reaction mixture is poured into ice cold water and extracted with ethyl acetate. The organic layer is dried over sodium sulfate, concentrated under vacuum, and the purification unit performs column chromatography to isolate compound 8.The reaction chamber facilitates deprotection of compound 8 to (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone hydrochloride (compound 9). Compound 8 is dissolved in 10 ml of 4 M HCl in 1,4-dioxane. The reaction is stirred at room temperature for 12 hours. The monitoring unit follows the course of the reaction by means of TLC with 60% ethyl acetate in hexane. After completion of the reaction, the reaction mixture is concentrated to dryness under vacuum. The crude product is triturated with 25 ml hexane to give an off-white solid identified as compound 9.The reaction chamber facilitates the condensation of compound 9 with various substituted 2-chloro-N-phenylacetamide derivatives (compounds 10a-l) to obtain the final target compounds (compounds 11a-l). A stirred solution of compound 9 in DMF is prepared and DIPEA is added at 0°C. The mixture is stirred at room temperature for 5 minutes, followed by addition of compound 10a-L. The reaction mixture is stirred at 60° C. for 8 hours. The monitor tracks progress by TLC using 5% methanol in dichloromethane (DCM), with visualization under UV light, iodine and ninhydrin. After completion, the reaction mixture is poured into ice cold water and extracted with ethyl acetate. The organic layer is dried over sodium sulfate, concentrated under vacuum, and the purification unit isolates compounds 11a-l by column chromatography. A secondary reaction chamber is available for the synthesis of substituted 2-chloro-N-phenylacetamide derivatives (compounds 10a-l). Substituted aniline is added to the chamber and dissolved in DMF. DIPEA is added at 0°C and the mixture is stirred at room temperature for 5 minutes. Subsequently, trifluoroacetyl chloride is added at 0°C and the reaction is stirred at room temperature for 3 to 10 hours. The monitoring unit tracks the course of the reaction by means of DC. After completion of the reaction, the mixture is poured into ice cold water and the precipitation unit collects the precipitate. The solid is filtered, washed with 50 ml cold water and triturated with 25 ml hexane to give the final substituted compound (10a-l).In one embodiment, all synthesized imidazo[2,1-b]thiazole derivatives are characterized by IR, 1 H-NMR, 13 C-NMR, and ESI-MS spectroscopy methods. In the IR spectra, the presence of a (-CN) stretch absorption band at about 2222 cm -1 is seen, while amide spectroscopy at about 1696 cm -1 The 1 H-NMR signal of 11a at δ 2.65 ppm indicates the presence of 2-CH 2groups of the piperazine ring. The signal at δ 3.29 confirms the presence of a -CH 2 group. The multiplet corresponding to δ 3.75-3.77 ppm indicates the presence of other 2-CH 2groups of the piperazine ring. Multiplet peaks at δ 7.26-7.28, 7.38-7.41 and 7.78-7.80 indicate the presence of a monosubstituted benzene ring. Two imidazo[2,1-b]thiazole-CH singlet peaks appear at δ 7.71 and 8.24, respectively. In addition, two doublets at δ 7.84-7.86 and 7.88-7.89 ppm correspond to the parasubstituted benzene ring. The singlet peak at δ 10.24 is due to the amide peak. In the 13 C-NMR spectrum of the imidazo[2,1-b]thiazole compounds 11a, two singlet signals are found at δ 105.62 and 110.50 ppm, respectively, owing to the two CH carbons of the imidazo[2,1-b]thiazole skeleton. The signals of the remaining three quaternary carbon atoms of the imidazo[2,1-b]thiazole backbone are δ 119.50 ppm, 124.84 ppm, 127.65 ppm and 134.44 ppm. For the amide carbon, the signal is lowest at δ 158.75 and 169.49, respectively. The -CN carbon is at δ116.93 ppm. The aromatic carbon exhibits intense peaks at 119.96 ppm, 125.30 ppm, 129.10 ppm and 133.68 ppm each having two carbon atoms. The characterization details of the individual compounds of the synthesized imidazo[2,1-b]thiazole derivatives are listed below:N-(4-Cyanophenyl)-2-(4-(6-phenylimidazo[2,1-blthiazole-3-carbonyl)piperazin-1-yl)acetamide (11a)Cream-coloured solid; melting point 181.2°C; IR (KBr, cm -1) vmax: 3276, 2922, 2222, 1696, 1632, 1384; 1H NMR (500 MHz, DMSO-d6) δ 2.6 (s, 4H, -CH2-), 3.29 (m, 2H, -CH2-), 3.75-3.77 (m, 4H, CH2), 7.27-7.28 (m, 1H, Ar-H), 7.38-7.41 (m, 2H, Ar-H), 7.71 (s, 1H, Ar-H), 7.78-7.80 (d, J=8 Hz, 2H, ArH), 7.84-7.86 (d, 2H, ArH), 7.99-8.04 (m, 2H, ArH), 7.88-7.89 (d, 2H, ArH), 8.24 (s, 1H, Ar-H); 13C NMR (100 MHz, DMSO-d6): δ 17.69, 21.52, 29.47, 30.57, 48.95, 52.86, 61.67, 105.62, 110.50, 116.93, 119.50, 119.96, 124.84, 125.30. 127.65, 129.10, 133.68, 134.44, 143.32, 146.81, 148.35, 158.75, 169.49; ESI-MS (m / z): 471.15 (M+1).N-(2,4-Dichlorophenyl)-2-(4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11b)Cream-coloured solid, melting point 201.4°C; IR (KBr, cm-1) v max: 3415, 3265, 2923, 1694, 1630, 1384; 1H NMR (500 MHz, DMSO-d6) δ 2.71 (s, 4H, -CH2-), 3.30 (s, 2H, -CH2-), 3.81 (m, 4H, CH2), 7.26-7.29 (m, 1H, Ar-H), 7.38-7.41 (m, 4H, Ar-H), 7.74 (s, 1H, Ar-H), 7.88-7.90 (d, J=8 Hz, 2H, ArH), 7.19-7.20 (d, 1H, ArH), 7.25 (s, 1H, ArH), 10.05 (s, 1H, Ar-H); 13C-NMR (100 MHz, DMSO-d6): δ 21.52, 52.96, 61.35, 110.50, 117.07, 120.80, 122.44, 124.83, 125.31, 127.65, 128.97, 129.11, 132.11, 134.45, 136.82, 146.82, 148.37, 158.87, 168.89; ESI-MS (m / z): 514.08 (M+1).N-(2-Ethyl-6-methylphenyl)-2-(4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11c)Cream-coloured solid; melting point 188.9 °C; IR (KBr, cm-1) v max: 3424, 3266, 2923, 2426, 1631, 1384; 1H-NMR (500 MHz, DMSO-d6) δ 1.09-1.12 (t, 3H, -CH3), 3.15 (s, 3H, -CH3), 2.68 (m, 4H, CH2), 3.23 (s, 2H, CH2), 3.79-3.81 (m, 4H, CH2), 7.08-7.10 (m, 2H, Ar-H), 7.13-7.14 (m, 1H, Ar-H), 7.25-7.28 (m, 2H, Ar-H), 7.72 (s, 1H, Ar -H), 7.88-7.90 (d, J=8 Hz, 2H, ArH), 8.25(s, 1H, Ar -H), 9.29(s, 1H, Ar -H); 13C-NMR (100 MHz, DMSO-d6): δ 15.03, 18.76, 24.93, 53.30, 61.30, 110.50, 116.94, 124.84, 125.31. 126.41, 127.26, 127.64, 128.12, 129.10, 134.46, 134.90, 136.11, 141.52, 146.81, 148.37, 158.77, 168.69; ESI-MS (m / z): 488.20 (M+1).N-(4-Fluoro-2-methoxyphenyl)-2-(4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11d)White solid; melting point 191.2°C; IR (KBr, cm-1) v max: 3305, 2921, 2426, 1683, 1632, 1384; 1H NMR (500 MHz, DMSO-d6) δ 2.64 (s, 4H, -CH2-), 3.26 (m, 2H, -CH2-), 3.76-3.78 (m, 4H, CH2), 6.91-6.94 (m, 1H, Ar-H), 7.25-7.28 (m, 1H, Ar-H), 7.38-7.44 (m, 4H, Ar-H), 7.71 (s, 1H, Ar-H), 7.88-7.89 (d, J=8 Hz, 2H, ArH), 8.24 (s, 1H, Ar-H), 10.17 (s, 1H, Ar-H); 13c nmr (100 mhz, DMSO-d6): δ 53.21, 56.17, 61.83, 98.70, 98.97, 106.83, 107.05, 109.54, 115.45, 120.59, 120.68, 123.04, 123.07, 124.44, 125.28, 127.65, 128.74, 133.70, 147.93, 148.31, 149.25, 149.35, 158.16, 159.30, 160.37, 166.99; ESI-MS (m / z): 494.16 (M+1).N-(3,5-difluorophenyl)-2-(4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11e)Tacky solid; IR (KBr, cm-1) v max: 3422, 3285, 2923, 2426, 1699, 1630, 1384; 1H NMR (500 MHz, DMSO-d6): δ 2.66 (s, 4H, -CH2-), 3.22 (s, 2H, -CH2-), 3.77-3.78 (m, 4H, CH2), 3.92 (s, 3H, -CH3), 6.75-6.79 (m, 1H, Ar-H), 7.01-7.04 (m, 1H, Ar-H), 7.26-7.29 (m, 1H, Ar-H), 7.38-7.41 (m, 2H, Ar-H), 7.73 (s, 1H, Ar-H), 7.88-7.90 (d, J=8 Hz, 2H, ArH), 8.10-8.13 (d, 1H, ArH), 8.26 (s, 1H, Ar-H), 9.58 (s, 1H, Ar-H), 13C-NMR (100 MHz, DMSO-d6): δ 53.38, 61.83, 99.83, 102.39, 102.69, 109.50, 115.54, 124.35, 125.29, 127.65, 128.74, 133.73, 139.28, 148.06, 148.33, 159.29, 162.11, 164.42167.48; ESI-MS (m / z): 482.14 (M+1).N-(4-Chloro-3-methylphenyl)-2-(4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11f)Pale yellow solid; melting point 192.8°C; IR (KBr, cm -1) v max: 3318, 2922, 2426, 1678, 1631, 1384; 1H NMR (500 MHz, CDCl3): δ 2.37 (s, 4H, -CH2-), 2.73-2.76 (m, 4H, -CH2-), 3.24 (s, 2H, -CH2-), 3.89-3.91 (m, 4H, CH2), 7.05 (m, 1H, Ar-H), 7.26-7.29 (m, 2H, Ar-H), 7.30-7.32 (m, 1H, Ar-H), 7.34-7.37 (m, 2H, Ar-H), 7.39-7.47 (m, 1H, Ar-H), 7.82-7.84 (d, 2H, ArH), 8.00 (s, 1H, ArH), 8.86 (s, 1H, Ar-H),; 13C-NMR (100 MHz, DMSO-d6): δ20.23, 29.69, 53.34, 61.85, 109.55, 115.57, 118.29, 121.86, 124.35, 125.28, 127.65, 128.74, 129.48, 129.77, 133.71, 135.70, 136.89, 147.95, 148.33, 159.23, 167.27,; ESI-MS (m / z): 494.13 (M+1).2-(4-(6-Phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)-N-(p-tolyl)acetamide (11 g) cream-coloured solid, melting point 200.4°C; IR (KBr, cm-1) v max: 3432, 3290, 2923, 2426, 1694, 1633, 1384; 1H NMR (500 MHz, CDCl3): δ 2.75-2.76 (m, 4H, -CH2-), 3.26 (m, 2H, -CH2-), 3.90 (m, 4H, CH2), 7.05 (m, 1H, Ar-H), 7.26-7.31 (m, 2H, Ar-H), 7.39-7.43 (m, 2H, Ar-H), 7.60-7.62 (d, J=8 Hz, 2H, ArH), 7.69-7.71 (d, 2H, ArH), 7.82-7.84 (m, 2H, ArH), 7.99 (s, 1H, Ar-H), 9.06 (s, 1H, Ar-H); 13C-NMR (100 MHz, CDCl3): δ 29.70, 53.37, 61.90, 109.54, 115.60, 119.17, 124.32, 125.30, 126.19, 126.39, 126.43, 127.66, 128.74, 133.71, 140.23, 147.99, 148.35, 159.26, 167.66; ESI-MS (m / z): 514.14 (M+1).N-(4-( tert-butyl)phenyl)-2-(4-(6-phenylimidazo[2,1- b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11 h)Cream-colored solid; melting point 201.3° C.; IR (KBr, cm -1) v max: 3454, 3305, 2957, 2426, 1680, 1633, 1384; 1H NMR (400 MHz, CDCl3): δ 1.31 (s, 9H, -CH3), 2.75 (s, 4H, -CH2-), 3.25 (m, 2H, -CH2-), 3.89 (m, 4H, CH2), 7.05 (s, 1H, Ar-H), 7.28-7.31 (m, 1H, Ar-H), 7.31-7.43 (m, 4H, Ar-H), 7.47-7.49 (d, 2H, Ar-H), 7.83-7.85 (d, J=8 Hz, 2H, ArH), 8.00 (s, 1H, Ar-H), 8.83 (s, 1H, Ar-H); 13C-NMR (100 MHz, CDCl3): δ 29.70, 31.35, 34.42, 53.33, 61.90, 109.55, 115.48, 119.43, 124.39, 125.28, 125.95, 127.61, 128.73, 133.79, 134.56, 147.69, 147.98, 148.34, 159.24, 167.16; ESI-MS (m / z): 502.22 (M+1).N-(3,5-Dichlorophenyl)-2-(4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11i)White solid; melting point 200.3°C; IR (KBr, cm-1) v max: 3440, 3283, 2923, 2426, 1697, 1626, 1384; 1H NMR (400 MHz, DMSO-d6) δ 2.74 (s, 4H, -CH2-), 3.25 (m, 2H, -CH2-), 3.91 (m, 4H, CH2), 7.05 (m, 1H, Ar-H), 7.30-7.32 (m, 2H, Ar-H), 7.39-7.43 (m, 3H, Ar-H), 7.73-7.74 (m, 2H, Ar-H), 7.82-7.84 (d, J=8 Hz, 2H, ArH), 8.00 (s, 1H, Ar-H), 8.97 (s, 1H, Ar-H); 13c nmr (100 mhz, DMSO-d6): δ 29.69, 30.57, 48.95, 53.32, 61.76, 109.53, 115.58, 121.01, 123.12, 124.33, 125.27, 127.64, 128.73, 129.89, 133.74, 139.37, 147.97, 148.32, 159.23, 167.54; ESI-MS (m / z): 514.8(M+1).N-(3-Cyanophenyl)-2-(4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11j)Tacky solid; (KBr, cm-1) v max: 3442, 3284, 2922, 2426, 1668, 1631, 1384; 1H-NMR (400 MHz, CDCl3): δ 2.68 (s, 4H, -CH2-), 3.20 (m, 2H, -CH2-), 3.84 (m, 4H, CH2), 7.03 (m, 1H, Ar-H), 7.24-7.27 (m, 1H, Ar-H), 7.35-7.43 (m, 4H, Ar-H), 7.75-7.80 (m, 3H, ArH), 7.98-8.00 (d, 2H, ArH), 9.14 (s, 1H, Ar-H); 13C-NMR (100 MHz, CDCl3): δ 31.46, 36.54, 53.28, 61.72, 109.64, 112.91, 115.72, 118.48, 122.65, 123.73, 124.30, 125.24, 127.66, 127.74, 127.83, 128.75, 130.00, 133.71, 138.19, 147.80, 148.33, 159.13, 162.60, 167.93; ESI-MS (m / z): 471.15 (M+1).N-(3-isopropoxyphenyl)-2-(4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)acetamide (11k)Pale yellow solid; melting point 200, 7°C; IR (KBr, cm-1) v max: 3298, 2958, 2426, 1680, 1632, 1384; 1H NMR (400 MHz, CDCl3): δ 1.25-1.27 (s, 6H, -CH3), 2.75-2.77 (m, 4H, -CH2-), 2.88-2.96 (m, 2H, -CH2-), 3.25 (m, 2H, -CH2-), 3.91 (m, 4H, CH2), 7.02-7.05 (m, 2H, Ar-H), 7.28-7.32 (m, 2H, Ar-H), 7.39-7.43 (m, 4H, Ar-H), 7.83-7.85 (m, 2H, Ar-H), 8.00 (s, 1H, ArH), 8.86 (s, 1H, Ar-H); 13C-NMR (100 MHz, CDCl3): δ 23.93, 29.69, 34.16, 53.33, 61.97, 109.54, 115.48, 117.14; ESI-MS (m / z): 504.20 (M+1).2-(4-(6- Phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazin-1-yl)-N-(thiazol-2-yl)acetamide (111)Tacky solid; IR (KBr, cm -1) v max: 3276 2922, 2222, 1696, 1632, 1384; 1H NMR (400 MHz, CDCl3): δ 2.72 (s, 4H, -CH2-), 3.35 (m, 2H, -CH2-), 3.90 (m, 4H, CH 2), 7.02-7.04 ( m, 2H, Ar-H), 7.26-7.30 (m, 1H, Ar-H), 7.38-7.46 (m, 1H, Ar-H), 7.81-7.83 (d, J=8 Hz, 2H, ArH), 7.99 (s, 1H, Ar-H); 13c nmr (100 mhz, DMSO-d6) δ 29.68, 53.43, 60.87, 109.56, 114.05, 115.55, 124.39, 125.28, 127.60, 128.72, 133.75, 137.20, 147.93, 148.35, 157.67, 159.16, 167.53; ESI-MS (m / z): 453.55 (M+1).In one embodiment, the synthesized series of piperazine-linked imidazo[2,1-b]thiazole derivatives [11(al)] is evaluated for their potential anti-cancer, anti-oxidation and anti-inflammatory activities. The synthesized compounds were extensively characterized and subjected to various biological tests to explore their pharmacological potential. Anti-cancer activity was evaluated using the sulforhodamine B test (SRB) on three human cancer cell lines: MCF-7 (breast cancer), COLO-205 (colon cancer) and K-562 (leukemia). The SRB assay measured the growth inhibition (GI50) of the compounds at various concentrations and determined their cytotoxic activity as compared to the reference drug adriamycin. In addition, the apoptosis-inducing ability of selected compounds in A549 lung cancer cells was evaluated using DAPI staining to identify nuclear condensation and apoptotic morphological changes. In addition, cell cycle arrest analysis was performed by flow cytometry using propidium iodide staining, particularly for compound 11a, to determine its effect on cell cycle distribution. For evaluation of antioxidant potential, the synthesized compounds were tested by a DPPH radical scavenger test, a FRAP test and a nitrogen oxide scavenger test. These tests measured the ability of the compounds to neutralize free radicals, reduce iron ions, and inhibit nitric oxide production, respectively, using gallic acid and ascorbic acid as standards. Anti-inflammatory potential was evaluated using the bovine serum albumin (BSA) antidenature assay, in which the inhibition of protein denaturation by the compounds was measured and compared to the standard drug diclofenac. An antiangiogenesis assay in the egg cell using the chorioallantoic membrane (CAM) of fertilized chicken eggs was carried out to examine the ability of the compounds (in particular 11a, 11e and 11f) to inhibit the formation of new blood vessels. This test was an important step in evaluating the antiangiogenic potential of the test compounds in a preclinical model. Finally, molecular docking studies were performed with the GOLD software to predict the binding affinity and interaction of the synthesized compounds with the BCR-ABL tyrosine kinase (PDB ID: 3IK3). Docking analysis compared the interactions of test compounds with those of the reference drug adriamycin, thus providing insight into the possible mechanism of cancer inhibitory activity at the molecular level. Together, these evaluations provide a comprehensive pharmacological profile of the synthesized imidazo[2,1-b]thiazole derivatives and demonstrate their potential as multifunctional therapeutics.The results of evaluation of the anti-cancer, anti-oxidative and anti-inflammatory properties of the produced imidazo[2,1-b]thiazole-N-phenylacetamidepiperazine derivatives (11a-111) gave the results described below.In anti-cancer screening, most synthesized compounds exhibited strong cytotoxic activity against the MCF-7 cell line (breast cancer) with GI50values below 10 μg / ml, comparable to the standard drug adriamycin. Exceptions were compounds 11d and 111 with their lower potency. In addition, compounds 11e, 11f, 11g and 11h exhibited high selectivity and activity against the K-562 cell line (leukemia), while all synthesized compounds exhibited the lowest cytotoxicity against the COLO-205 cell line (colon cancer). Anti-inflammatory activity was evaluated by BSA protein antidenature assay. Compounds 11a, 11c, 11d, 11e, 11f, 11i, 11j and 11k exhibited significant activity, indicating good anti-inflammatory potential. The compounds also exhibited remarkable antioxidant activity, which was confirmed by DPPH, FRAP and nitric oxide capture assays. Of all, compounds 11a, 11e and 11f with their multiple targeting biological profiles have consistently proved to be the most promising. In addition, molecular docking studies revealed strong binding affinities of 11a, 11e and 11f to the active site of BCR-ABL tyrosine kinase (PDB ID: 3IK3), with docking poses and interactions similar to those of the co-crystallized ligand (AP24534). These compounds also showed favourable ADMET profiles, including good absorption, distribution and metabolism with reduced risks of cardiotoxicity and liver toxicity compared to AP24534. Mechanistic evaluations also supported their pro-apoptotic potential, with DAPI staining showing clear apoptotic characteristics in A549 cells treated with compounds 11a and 11f. The in-ova CAM assay showed compound 11a to have strong antiangiogenic activity, indicating low toxicity and effective inhibition of blood vessel formation. In addition, cell cycle analysis confirmed that compound 11a induced a G2 / M phase arrest, a characteristic of antiproliferative activity. Compounds 11a, 11e and 11f have proven to be promising multifunctional guide molecules which combine potent anti-cancer, anti-inflammatory and anti-oxidative properties with a favourable safety profile. Their selective action, particularly against breast cancer and leukaemia cell lines, makes them attractive candidates for progression to effective cancer medications.The drawings and the foregoing description show examples of embodiments. Those skilled in the art will appreciate that one or more of the described elements may well be combined into a single functional element. Alternatively, certain elements may be divided into multiple functional elements. Elements of one embodiment may be added to another embodiment. For example, the order of the processes described herein may be changed and is not limited to the manner described herein. Moreover, the actions of a flow chart need not be performed in the order shown; nor do all actions necessarily need to be performed. Also, actions that are not dependent on other actions may be performed in parallel with the other actions. The scope of the embodiments is by no means limited by these specific examples. Numerous variations, whether or not explicitly stated in the specification, such as differences in structure, dimensions, and material use, are possible. The scope of the embodiments is at least as broad as recited in the following claims.Advantages, other advantages and solutions to problems have been described above with reference to specific embodiments. However, the advantages, merits, solutions to problems and any components that may result in an advantage, merit or solution being introduced or enhanced are not to be understood as critical, required or essential features or components of individual or all claims.REFERENCES100 A system for preparing piperazine-linked imidazo[2,1-B]thiazole derivatives as anti-cancer, antioxidant and anti-inflammatory agents. 102 Reaction chamber 102 aSecondary reaction chamber 104 Heater 106 Cooling unit 108 Agitator 110 Solvent supply system 112 Reagent supply system 114 Cleaning unit 116 Sampling unit 118 Precipitation unit 120 Monitoring unit

Claims

A system (100) for preparing piperazine-linked imidazo[2,1-b]thiazole derivatives as anti-cancer agents, as an antioxidant, and as an anti-inflammatory agent, comprising: a) a reaction chamber (102) configured to allow for multiple chemical reactions for synthesizing piperazine-linked imidazo[2,1-b]thiazole derivatives; b) a heating unit (104) operatively connected to the reaction chamber (102) and configured to provide controlled temperature conditions ranging from room temperature to reflux temperature; c) a cooling unit (106) operatively connected to the reaction chamber (102) and configured to maintain cooling temperatures during operation of the reaction chamber (102); d) a stirring unit (108) operatively connected to the reaction chamber (102) and configured to provide continuous mixing of the reaction components; e) a solvent supply system (110) configured to supply reaction solvent to the reaction chamber (102); f) a reagent supply system (112) configured to supply reaction reagents to the reaction chamber (102); g) a purification unit (114) configured to purify the synthesized piperazine-linked imidazo[2,1-b]thiazole derivatives by column chromatography; and h) a sample collection unit (116) configured to collect and store the resulting product after each reaction performed in the reaction chamber, wherein the sample collection unit (116) is connected to a supply system configured to supply the collected sample back to the reaction chamber for subsequent reactions.The system (100) of claim 1, wherein the reaction chamber (102) is configured to facilitate the synthesis of ethyl 2-aminothiazole-4-carboxylate (Compound 3), wherein the reaction between ethyl 3-bromo-2-oxopropanoate and thiourea is performed at 100°C for 3 hours to produce ethyl 2-aminothiazole-4-carboxylate.The system (100) of claim 1, wherein the reaction chamber (102) is further configured to facilitate the reaction between ethyl 2-aminothiazole-4-carboxylate (Compound 3) and 2-bromo-1-phenylethanone in the presence of sodium bicarbonate at reflux temperature for 8 hours to synthesize ethyl 6-phenylimidazo[2,1-b]thiazole-3-carboxylate (Compound 5).The system (100) of claim 1, wherein the reaction chamber (102) is further configured to allow a reaction for converting ethyl 6-phenylimidazo[2,1-b]thiazole-3-carboxylate (Compound 5) to 6-phenylimidazo[2,1-b]thiazole-3-carboxylic acid (Compound 6) using lithium hydroxide at room temperature for 12 hours, wherein ethyl 6-phenylimidazo[2,1-b]thiazole-3-carboxylate (Compound 5) is hydrolyzed in the presence of THF, methanol, water, and lithium hydroxide.The system (100) of claim 1, wherein the reaction chamber (102) is further configured to allow a reaction between 6-phenylimidazo[2,1-b]thiazole-3-carboxylic acid (Compound 6) and tert-butylpiperazine-1-carboxylate (Compound 7) in the presence of DMF, HATU, and DIPEA to synthesize tert-butyl 4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazine-1-carboxylate (Compound 8).The system (100) of claim 1, wherein the reaction chamber (102) is further configured to allow a reaction to convert tert-butyl 4-(6-phenylimidazo[2,1-b]thiazole-3-carbonyl)piperazine-1-carboxylate (Compound 8) to (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone hydrochloride (Compound 9) using HCl in 1,4-dioxane at room temperature for 12 hours.The system (100) of claim 1, further comprising a secondary reaction chamber (102a) configured to facilitate the synthesis of substituted 2-chloro-N-phenylacetamide compounds (10 (al)) by the reaction of trifluoroacetyl chloride with substituted anilines in the presence of DMF and DIPEA.The system (100) of claim 1, wherein the reaction chamber (102) is further configured to facilitate synthesis of the final target compound by allowing a reaction between (6-phenylimidazo[2,1-b]thiazol-3-yl)(piperazin-1-yl)methanone hydrochloride (compound 9) and substituted 2-chloro-N-phenylacetamide compounds (10 (al)) at 60°C for 8 hours.The system (100) of claim 1, further comprising a precipitation unit (118) coupled to the reaction chamber (102), a cleaning unit (114), and a sample collection unit (116) configured to isolate reaction products by adding reaction mixtures to ice-cold water and collecting the resulting precipitates.The system (100) of claim 1, further comprising a monitoring unit (120) connected to the reaction chamber (102) and configured to track the course of the reaction using thin layer chromatography (TLC) with visualization under UV light, iodine, and ninhydrin.

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