Ethyl chloroacetate

Chemicals: chemicals of this work (4-(N,N-dimethylaminobenzaldhyde, Cyanothioacetamide, Piperidine, Methyl iodide, Ethyl Chloroacetate, 2-Chloroacetamide, Chloroacetonitrile or N-aryl-2-Chloroacetamides, Ethanol, Sodium acetate.3H₂O, Sodium carbonate) were purchased from Sigma Aldrich Co.
Instrumentations: Melting points were determined on a Gallan-Kamp apparatus and are uncorrected. The purity of the compounds was ensured by TLC and the spectroscopic analysis.
IR spectra were recorded on a Shimadzu 470 IR-spectrophotometer (KBr; ν_max in cm⁻¹). The ¹H and ¹³C NMR spectra were recorded on Varian A5 500 MHz spectrometer using DMSO-d₆ as a solvent and tetramethylsilane (TMS) as an internal reference. Coupling constants (J values) are given in Hertz (Hz). Elemental analyses were performed on a Perkin Elmer 2400 LS Series CHN/O analyzer.
Cell lines: The in vitro human breast cancerous cell line (MCF7), lung cancerous  cell lines (A549) and normal cell lines were purchased from Serum and Vaccine formulation in Cairo-Egypt.
Molecular docking: Molecular docking studies were performed in (I Mole Lab for bioinformatics, Cairo, Egypt).
Softwares: The biological data was analyzed and plot by Graphpad prism, Cell qust, ANOVA, Origin Lab, AutoDock Vina 1.1.2, Mestrenova and Excel software.

All melting points are uncorrected. Compounds prepared by different procedures were characterized by mixed melting points, TLC, and IR. IR spectra (KBr) were recorded on a Fourier transform infrared spectrophotometer (IRAffinity-1S from Shimadzu Corporation). NMR spectra were measured with a Varian Gemini 300 spectrometer (300 MHz ¹H NMR), and chemical shifts are given in ppm from TMS. ¹³C NMR spectra were recorded with a Varian Mercury 300 spectrometer (300 MHz ¹H NMR, 75 MHz ¹³C NMR). The ¹H NMR samples were measured in 5 mm NMR tubes with a concentration of about 20 mg of the product in 0.6 ml of solvent (DMSO-d₆) and about 75–100 mg of the product in about 0.6 ml of solvent for ¹³C NMR. Mass spectra were recorded on DI Analysis Shimadzu QP-2010 Plus. Elemental analyses were carried out at the Microanalytical Center, Cairo University. 1,4-Cyclohexanedione, ethyl chloroacetate, chloroacetic acid, ethyl bromoacetate, α-chloroacetylacetone, and thiosemicarbazide were used as purchased from Aldrich. The starting materials 4a–g were prepared as reported in the literature (Eweiss and Osman, 1980 (link); Farag and Algarib, 1988 (link)).

Triazol was purchased from Sigma Chemical Co. (St. Louis, MO). The Girard
reagent GTr was synthetized dissolving Triazol (7.5 x 10^-3 moles)
in ethanol (20 mL) and mixed with ethyl chloroacetate (Aldrich) (9.4 x
10^-3 moles). The reaction mixture was stirred in reflux at
90°C for 72 hours and then was settled on ice and mixed with hydrazine
hydrate (2.9 x 10^-2 moles) under agitation for 20 minutes. The
solvent was evaporated in an ice bath, the residue was recrystallized in
methanol, and solid purified by column chromatography on alumina.