Apparatus and composition for the synthesis of acetamide derivatives as antioxidants and antimicrobials in two pots

Abstract

A composition for the synthesis of a cetamide derivative as an antioxidant and antimicrobial agent, the composition comprising: 8-10 g 2,3-Dimethylaniline; 5-6 ml chloroacetyl chloride; 95-105 ml toluene; and 1-2 g 2-amino-4,6-dihydroxypyrimidine.

Description

FIELD OF THE INVENTION

[0001] The present disclosure relates to a two-pot synthesis method and a composition for synthesizing derivative compounds. More specifically, the present invention discloses a two-pot synthesis method and a composition for synthesizing acetamide derivatives as antioxidants and antimicrobials. BACKGROUND OF THE INVENTION

[0002] A broad spectrum of microorganisms, including bacteria, viruses, protozoa, and fungi, are increasingly developing resistance to drugs used to treat infections, posing a significant global challenge in the treatment of infectious diseases. The World Health Organization (WHO) recognizes growing resistance to antimicrobial drugs as the most serious threat to human health. Multidrug-resistant microorganisms have reached critical levels worldwide, underscoring the urgent need for antimicrobials with increased efficacy.

[0003] Cancer remains a serious global health problem and one of the leading causes of death. Researchers have tirelessly searched for effective clinical approaches and new anti-cancer agents over the past decades. Inflammatory cells, cytokines, chemokines, and enzymes play a role in promoting cancer development from inflammation. This suggests a fascinating connection between cancer and inflammation.

[0004] Biochemical studies of structures containing nitrogen heteroatoms have demonstrated their ability to uncouple oxidative phosphorylation, inhibit mucopolysaccharide biosynthesis, and, in particular, inhibit prostaglandin biosynthesis at the COX stage. This contributes to their anti-inflammatory and analgesic effects. Tissue inflammation can result from an infection that promotes bacterial growth and leads to chronic inflammation. A drug with antibacterial and anti-inflammatory properties can act synergistically, combating both inflammation and infection.

[0005] Nitrogen-containing heterocyclic pyrimidine derivatives are important in medicinal chemistry due to their presence in nucleic acids. Literature reviews show that compounds containing a pyrimidine core exhibit diverse pharmacological activities, including antibacterial, antifungal, antileishmanial, anti-mycobacterial, anti-inflammatory, anticancer, antiviral, antituberculous, antimalarial, antioxidant, CNS depressant, and calcium channel blocking effects. Interest in pyrimidine derivatives, driven by their extensive clinical applications, has led to the preparation and evaluation of numerous molecules as potent anti-inflammatory and anticancer agents. This study highlights the importance of pyrimidine as a crucial pharmacophore in the discovery of new drugs.

[0006] Therefore, to meet the above requirements, there is a need to develop a two-pot synthesis device and composition to synthesize an acetamide derivative by combining different pharmacophores in heterocyclic structures to obtain compounds with potential antioxidant and antimicrobial activity.

[0007] The technical advances disclosed by the present invention overcome the limitations and disadvantages of existing and conventional systems and methods. SUMMARY OF THE INVENTION

[0008] The present disclosure relates to a two-pot synthesis process and composition for the synthesis and characterization of a drug molecule - a pyrimidine acetamide derivative containing a pyrimidine residue with improved pharmacological properties.

[0009] The present disclosure describes a composition for synthesizing an acetamide derivative as an antioxidant and antimicrobial agent, the composition comprising: 8-10 g of 2,3-dimethylaniline, 5-6 ml of chloroacetyl chloride, 95-105 ml of toluene, and 1-2 g of 2-amino-4,6-dihydroxypyrimidine.

[0010] The present disclosure is intended to provide a two-pot synthesis apparatus for synthesizing an acetamide derivative as an antioxidant and antimicrobial agent. The apparatus comprises a flask configured to prepare 2-chloro-N-(2,3-dimethylphenyl)acetamide by mixing 9.0 g of 2,3-dimethylaniline with 5.24 mL of chloroacetyl chloride along with 100 mL of toluene; and a round-bottom flask filled with 1.50 g of 2-chloro-N-(2,3-dimethylphenyl)acetamide, which has been reacted with 1.5 g of 2-amino-4,6-dihydroxypyrimidine in the presence of sodium hydride with dimethylformamide (DMF) to form 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide residue.

[0011] An object of the present disclosure is to provide a two-pot synthetic route for the synthesis of acetamide derivatives as an antioxidant and antimicrobial agent;

[0012] Another objective of the present disclosure is to evaluate the pharmacological potential of synthesized 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide by antioxidant and antimicrobial activities in vitro.

[0013] A further aim of the present disclosure is the use of the intermediate 2-chloro-N-(2,3-dimethylphenyl)acetamide obtained in the first step for the synthesis of acetamide derivatives with a pyrimidine skeleton.

[0014] Another objective of the present disclosure is to determine efficacy as an effective drug molecule against fungal infections, diabetes, inflammation and cancer.

[0015] To further clarify the advantages and features of the present disclosure, a more detailed description of the invention will be given with reference to specific embodiments thereof illustrated in the accompanying drawings. It should be noted that these drawings represent only typical embodiments of the invention and are therefore not to be considered limiting its scope. The invention will be described and explained in additional detail and in greater detail with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE CHARACTERS

[0016] These and other features, aspects, and advantages of the present disclosure will be better understood when the following detailed description is read with reference to the accompanying drawings, in which like characters represent like parts throughout. Fig. 1 shows a block diagram of a two-pot synthesis apparatus for synthesizing an acetamide derivative as an antioxidant and antimicrobial agent according to an embodiment of the present disclosure; Fig. Figure 2 shows a schematic synthesis of 2-chloro-N-(2,3-dimethylphenyl)acetamide according to an embodiment of the present disclosure; and Fig. Figure 3 illustrates a structure of 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide according to one embodiment of the present disclosure.

[0017] Furthermore, those skilled in the art will appreciate that elements in the drawings are shown for convenience and may not necessarily be drawn to scale. For example, the flowcharts illustrate the method by key steps to enhance understanding of aspects of the present disclosure. Moreover, with respect to device construction, one or more components of the device may have been represented in the drawings by conventional symbols, and the drawings may show only those specific details relevant to understanding embodiments of the present disclosure in order not to clutter the drawings with details that would be readily apparent to those skilled in the art who would benefit from the description herein. DETAILED DESCRIPTION:

[0018] To facilitate an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the drawings and described in specific language. It is to be understood, however, that this is not intended to limit the scope of the invention, as such changes and further modifications to the illustrated system and such further applications of the principles of the invention as illustrated therein are contemplated as would normally occur to one skilled in the art to which the invention pertains.

[0019] It will be understood by one 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.

[0020] References in 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. Therefore, occurrences of the phrase "in one embodiment," "in another embodiment," and similar language throughout this specification may or may not all refer to the same embodiment.

[0021] The terms "comprises," "comprising," or other variations thereof are intended to cover non-exclusive inclusion, such that a process or method comprising a list of steps not only includes those steps, but may also include other steps not expressly listed or inherent in such process or method. Likewise, one or more devices or subsystems or elements or structures or components preceded by "comprises...a" does not preclude, without further limitation, the existence of other devices or other subsystems or other elements or other structures or other components or additional devices or additional subsystems or additional elements or additional structures or additional components.

[0022] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The system, methods, and examples provided herein are for illustrative purposes only and should not be considered limiting.

[0023] Embodiments of the present disclosure will be described in detail below with reference to the accompanying drawings.

[0024] A composition for synthesizing an acetamide derivative as an antioxidant and antimicrobial agent, the composition comprising: 8-10 g of 2,3-dimethylaniline; 5-6 ml of chloroacetyl chloride; 95-105 ml of toluene; and 1-2 g of 2-amino-4,6-dihydroxypyrimidine. Specifically, the composition comprises: 9 g of 2,3-dimethylaniline; 5.24 ml of chloroacetyl chloride; 100 ml of toluene; and 1 g of 2-amino-4,6-dihydroxypyrimidine.

[0025] Fig. 1 shows a block diagram of a two-pot synthesis apparatus (100) for synthesizing an acetamide derivative as an antioxidant and antimicrobial agent, the apparatus (100) comprising: a flask (102), a dropping funnel (104), a round-bottom flask (106), a condenser (108), and a purification unit (110).

[0026] The flask (102) is used to prepare 2-chloro-N-(2,3-dimethylphenyl)acetamide by mixing 8 - 10 g of 2,3-dimethylaniline with 5 - 6 ml of chloroacetyl chloride and 95 - 105 ml of toluene.

[0027] A dropping funnel (104) is attached to the opening of the flask (102) to add dropwise a mixture of 5-6 ml of chloroacetyl chloride together with 95-105 ml of toluene into the flask (102) containing 2,3-dimethylaniline, wherein a condenser (108) is connected to the flask (102) to introduce 2,3-dimethylaniline into the flask (102).

[0028] The round-bottomed flask (106) is operatively connected to the flask (102) charged with 1-2 g of 2-chloro-N-(2,3-dimethylphenyl)acetamide, which reacts with 1-2 g of 2-amino-4,6-dihydroxypyrimidine in the presence of sodium hydride with dimethylformamide (DMF) to form the 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide residue.

[0029] The flask (102) is heated under reflux for 6 hours after mixing 2,3-dimethylaniline with chloroacetyl chloride and toluene to obtain the intermediate 2-chloro-N-(2,3-dimethylphenyl)acetamide.

[0030] The purification unit (110) is operatively connected to the round-bottom flask (106) to purify the 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide residue by the silica gel column chromatography method and obtain the pure product of the acetamide derivative.

[0031] In an exemplary embodiment, the flask (102) is used to prepare 2-chloro-N-(2,3-dimethylphenyl)acetamide by mixing 9 g of 2,3-dimethylaniline with 5.24 ml of chloroacetyl chloride together with 100 ml of toluene, wherein a dropping funnel (104) is attached to the opening of the flask (102) to add dropwise 5.24 ml of chloroacetyl chloride together with 95-105 ml of toluene into the flask (102) containing 2,3-dimethylaniline.

[0032] The round-bottomed flask (106) is operatively connected to the flask (102) which is charged with 1.5 g of 2-chloro-N-(2,3-dimethylphenyl)acetamide (intermediate prepared in the first step) and reacted with 1-2 g of 2-amino-4,6-dihydroxypyrimidine in the presence of sodium hydride with dimethylformamide (DMF) to form the 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide residue (final product).

[0033] In one embodiment, thin-layer chromatography (TLC) is performed using F254 plates (silica gel coated with a fluorescent indicator), and column chromatography is performed using silica gel 60 (0.063-0.2 mm). Using potassium bromide (KBr) pellets, infrared (IR) spectra are measured using a 470 Fourier transform spectrophotometer. Nuclear magnetic resonance (1H-NMR) spectra are recorded using a 400 MHz spectrophotometer, with chemical shifts expressed in ppm, using tetramethylsilane (TMS) as an internal standard. Electrospray ionization mass spectrometry (ESI-MS) spectra are recorded using a high-performance liquid chromatography (HP-LC) system, and a 6320D spectrophotometer is used to detect 2,2-diphenyl-1-picrylhydrazyl (DPPH).

[0034] Fig. Figure 2 shows the structure of 2-chloro-N-(2,3-dimethylphenyl)acetamide and the schematic synthesis of 2-chloro-N-(2,3-dimethylphenyl)acetamide. 9.0 g of 2,3-dimethylaniline is added to a flask equipped with a condenser and a dropping funnel. Then, 5.24 ml of chloroacetyl chloride along with 100 ml of toluene is added dropwise. The flask is refluxed for 6 hours to obtain the intermediate 2-chloro-N-(2,3-dimethylphenyl)acetamide.

[0035] Fig.Figure 3 shows a structure of 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide, where a round-bottomed flask is charged with 1.50 g of the intermediate, i.e., 2-chloro-N-(2,3-dimethylphenyl)acetamide. This intermediate is reacted with 1.5 g of 2-amino-4,6-dihydroxypyrimidine in the presence of sodium hydride and dimethylformamide (DMF). The residue is purified by silica gel column chromatography to afford the pure product. The final product, 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide, is obtained in good yield.

[0036] In one embodiment, the presence of a hydroxyl group and an amine group is observed, along with the presence of a CH stretch of the alkane group (methyl group). The presence of a nitrogen-bonded carbonyl group (amide group) is observed. The peak corresponds to a phenylalkyl ether bond.

[0037] In one embodiment, the chemical structure of the synthesized molecule is then determined using various analytical techniques such as 1H NMR, FT-IR, and HPLC-MS. 1H NMR results revealed characteristic peaks of the functional groups present in the molecules. FT-IR studies also confirmed the presence of the prominent functional groups. Furthermore, the mass spectrometer showed a specific peak in the positive mode of the spectra, which confirmed the molecular weight of the molecule with the calculated one.

[0038] In one embodiment, biological activities, i.e., antioxidant and antimicrobial, are performed to evaluate the pharmacological potential of 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide.

[0039] The antioxidant activity of 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide is determined using the DPPH assay. Different concentrations of the sample are added to test tubes containing 0.1 ml of methanol. Then, 3 ml of DPPH solution with a preset absorbance of 1 is added and incubated for 15 minutes in the dark. The absorbance is measured at 517 nm with methanol, using gallic acid as the reference standard. Inhibition percentage=[control antibody−sample antibody / control antibody]*100 Antimicrobial activity.

[0040] The antimicrobial efficacy of 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide is investigated using the agar diffusion method against S. aureus, P. aeruginosa, and C. albicans, with tetracycline serving as a control. The compound and tetracycline (10 mg each) are diluted in 1 ml of DMSO (dimethyl sulfoxide). Aliquots of the sample and control are prepared by pipetting 10 µl, 20 µl, 30 µl, and 40 µl, respectively, and made up to 50 µl with DMSO. After autoclaving at 121°C for 15 minutes, the bacterial plates are incubated for 24 hours at 37°C and the fungal plates for 72 hours at 25°C. The inhibition zone is then measured in millimeters.

[0041] The drawings and the foregoing description provide examples of embodiments. Those skilled in the art will recognize that one or more of the described elements may well be combined into a single functional element. Alternatively, certain elements may be separated into multiple functional elements. Elements from one embodiment may be added to another embodiment. For example, orders of the processes described herein may be changed and are not limited to the manner described herein. Furthermore, the actions of any flowchart need not be implemented in the order shown; nor do all actions necessarily need to be performed. Also, those actions that are not dependent on other actions may be performed in parallel with the other actions. The scope of the embodiments is in no way limited by these specific examples.Numerous variations, whether explicitly stated in the specification or not, such as differences in structure, dimensions, and use of materials, are possible. The scope of the embodiments is at least as broad as indicated in the following claims.

[0042] Advantages, other benefits, and solutions to problems have been described above with reference to specific embodiments. However, the advantages, benefits, solutions to problems, and any components that may cause an advantage or solution to occur or become more apparent are not to be construed as a critical, required, or essential feature or component of any or all of the claims. REFERENCES 100 A two-pot synthesis process for the preparation of an acetamide derivative as an antioxidant and antimicrobial agent. 102 bottles 104 dropping funnels 106 round-bottom flasks 108 Capacitor 110 cleaning unit

Claims

[1] A composition for synthesizing a cetamide derivative as an antioxidant and antimicrobial agent, the composition comprising: 8-10 g 2,3-dimethylaniline; 5-6 ml chloroacetyl chloride; 95-105 ml toluene; and 1-2 g 2-amino-4,6-dihydroxypyrimidine. [2] Composition according to claim 1, wherein the composition comprises in particular: 9 g 2,3-dimethylaniline; 5.24 ml chloroacetyl chloride; 100 ml toluene; and 1.5 g 2-amino-4,6-dihydroxypyrimidine. [3] A two-pot synthesis apparatus (100) for synthesizing an acetamide derivative as an antioxidant and antimicrobial agent, the apparatus (100) comprising: a flask (102) for preparing 2-chloro-N-(2,3-dimethylphenyl)acetamide by mixing 8-10 g of 2,3-dimethylaniline with 5-6 ml of chloroacetyl chloride and 95-105 ml of toluene; a dropping funnel (104) attached to the opening of the flask (102) for adding 5-6 ml of chloroacetyl chloride together with 95-105 ml of toluene dropwise into the flask (102) filled with 2,3-dimethylaniline; and a round-bottomed flask (106) operatively connected to the flask (102) charged with 1-2 g of 2-chloro-N-(2,3-dimethylphenyl)acetamide which has been reacted with 1-2 g of 2-amino-4,6-dihydroxypyrimidine in the presence of sodium hydride with dimethylformamide (DMF) to form the residue 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide. [4] The apparatus of claim 3, wherein a cooler (108) is connected to the flask (102) for introducing 2,3-dimethylaniline into the flask (102). [5] The apparatus of claim 3, wherein after mixing the 2,3-dimethylaniline with chloroacetyl chloride and toluene, the flask (102) is heated under reflux for 6 hours to obtain the intermediate 2-chloro-N-(2,3-dimethylphenyl)acetamide. [6] The apparatus of claim 3, wherein a purification unit (110) is operatively connected to the round-bottomed flask (106) for purifying the 2-[(4,6-dihydroxypyrimidin-2-yl)amino]-N-(2,3-dimethylphenyl)acetamide residue by the technique of silica gel column chromatography to obtain the pure product.

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