Compound, preparation method and application thereof, and pharmaceutical composition comprising compound and N-oxalyl glycyl glycine ethyl ester

Abstract

The invention relates to the technical field of medicinal chemistry, in particular to a compound as well as a preparation method and application thereof, and a pharmaceutical composition comprising the compound and N-acyl glycyl glycine ethyl ester. The structural general formula of the compound is shown in the specification, wherein R1 is selected from any one of a hydroxyl group, a methoxy group, an ethyoxyl group, a propoxy group, a benzyloxy group and-NHCH2CONHCH2COOCH2CH3, and R2 is selected from any one of a hydroxyl group, a methoxy group, an ethyoxyl group, a propoxy group, a benzyloxy group and- R2 is selected from any one of hydroxyl, methoxyl and ethoxyl; and R1 and R2 are not hydroxyl at the same time. On the basis of N-oxalic acid-acyl glycyl glycine ethyl ester, research is continued, impurities in N-oxalic acid-acyl glycyl glycine ethyl ester are separated and synthesized, and the compound obtained through separation serves as a reference substance to be applied to quality control of N-oxalic acid-acyl glycyl glycine ethyl ester. And the quality and the safety of the N-oxalic acid-acyl glycyl glycine ethyl ester can be evaluated more accurately.

Description

Technical Field

[0001] This application relates to the technical field of medicinal chemistry, and more particularly to a compound, its preparation method and application, and pharmaceutical compositions including the compound and N-oxaloylglycylglycine ethyl ester. Background Technology

[0002] N-oxoylglycine ethyl ester is an N-oxoyl dipeptide derivative that reduces the delta and sigma indices of brain waves and increases the theta index, and can be used as a wakefulness-promoting drug.

[0003] Patent CN101747226A discloses a specific synthetic method for N-oxaloylglycylglycine ethyl ester as an N-oxaloyl dipeptide derivative, and also discloses its awakening experimental data in adult rabbits. The synthetic method involves preparing benzyl oxalate monomethyl ester monoacyl chloride from benzyl alcohol and oxalyl chloride as starting materials, then condensing it with glycylglycine ethyl ester hydrochloride in triethylamine to obtain benzyloxyoxaloylglycylglycine ethyl ester, followed by palladium-catalyzed hydrogenation to obtain N-oxaloylglycylglycine ethyl ester. The specific synthetic process is shown below:

[0004]

[0005] In order to better control the quality of N-oxoylglycylglycine ethyl ester, it is necessary to study its impurities and analogues and control them within a safe and reasonable range to ensure the quality and safety of the drug. Summary of the Invention

[0006] This application provides a compound, its preparation method, and its application, including a pharmaceutical composition comprising the compound and N-oxaloylglycylglycine ethyl ester. Based on the pharmaceutical compound N-oxaloylglycylglycine ethyl ester, this application further studies the compound by isolating and synthesizing its impurities. The compound obtained in this application is used as a reference standard for the quality control of N-oxaloylglycylglycine ethyl ester, enabling a more accurate assessment of its quality and safety.

[0007] In a first aspect, this application provides a compound, which adopts the following technical solution:

[0008] A compound having the following general structural formula:

[0009]

[0010] Wherein, R1 is selected from any one of hydroxyl, methoxy, ethoxy, propoxy, benzyloxy, and -NHCH2CONHCH2COOCH2CH3; R2 is selected from any one of hydroxyl, methoxy, and ethoxy; and R1 and R2 are not both hydroxyl, nor are they both benzyloxy and ethoxy.

[0011] Optionally, the compounds include compound B, compound C, compound D, compound E, compound F, compound G, compound H, compound I, and compound J.

[0012] In one specific embodiment, when R1 is -OCH2CH3 and R2 is -OCH2CH3, the compound is compound B.

[0013] The structural formula of compound B is as follows:

[0014] In one specific embodiment, when R1 is -NHCH2CONHCH2COOCH2CH3 and R2 is -OCH2CH3, the compound is compound D.

[0015] The structural formula of compound D is as follows:

[0016] In one specific embodiment, when R1 is -OCH2CH3 and R2 is -OH, the compound is compound E.

[0017] The structural formula of compound E is as follows:

[0018] In one specific embodiment, when R1 is -OCH3 and R2 is -OH, the compound is compound F.

[0019] The structural formula of compound F is as follows:

[0020] In one specific embodiment, when R1 is -OCH3 and R2 is -OCH3, the compound is compound G.

[0021] The structural formula of compound G is as follows:

[0022] In one specific embodiment, when R1 is -OCH3 and R2 is -OCH2CH3, the compound is compound H.

[0023] The structural formula of compound H is as follows:

[0024] In one specific embodiment, when R1 is -OCH2CH3 and R2 is -OCH3, the compound is compound I.

[0025] The structural formula of compound I is as follows:

[0026] In one specific embodiment, when R1 is -OCH2CH2CH3 and R2 is -OCH2CH3, the compound is compound J.

[0027] The structural formula of compound J is as follows:

[0028] Optionally, the preparation methods of compounds B and H are as follows: oxaloyl chloride monoethyl ester or oxaloyl chloride monomethyl ester is obtained by condensation with glycyl glycinate ethyl ester hydrochloride under alkaline conditions.

[0029] In the preparation methods of compounds B and H:

[0030] Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran. In one specific embodiment, the selected reaction solvent is dichloromethane.

[0031] Optionally, the alkaline condition is any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. In one specific embodiment, the alkaline condition is triethylamine.

[0032] Optionally, in the preparation method, the molar ratio of oxaloyl chloride monomethyl(ethyl) ester: glycyl glycinate hydrochloride: triethylamine is 1:(1-1.5):(2-3). In one specific embodiment, the molar ratio is 1:1:2.

[0033] Optionally, the extraction solvent used in the preparation method is any one or more of ethanol, ethyl acetate, methyl acetate, acetone, and butyl acetate. In one specific embodiment, the extraction solvent used is a mixture of ethanol and ethyl acetate.

[0034] Optionally, the purification and recrystallization method in the preparation method may use any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile. In one specific embodiment, anhydrous ethanol may be used as the purification and recrystallization method.

[0035] Optionally, the preparation method of compound D is as follows: oxalyl chloride and glycyl glycine ethyl ester hydrochloride are condensed under alkaline conditions.

[0036] In the preparation method of compound D:

[0037] Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran. In one specific embodiment, the selected reaction solvent is dichloromethane.

[0038] Optionally, the alkaline condition is any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. In one specific embodiment, the alkaline condition is triethylamine.

[0039] Optionally, the preliminary purification method in the preparation method involves dissolving the ester-soluble compound in ethyl acetate, methyl acetate, acetone, or butyl acetate, and then dissolving the water-soluble compound in water at 60-90°C to obtain the crude product. In one specific embodiment, the preliminary purification method involves dissolving the ester-soluble compound in ethyl acetate, then dissolving the water-soluble compound in water at 70°C, with the remaining solid being the crude product.

[0040] Optionally, the further purification method in the preparation method involves thermal dissolution of DMF or DMSO, decolorization and filtration with activated carbon, addition to water to precipitate the solid, and then slurrying with anhydrous ethanol, methanol, or isopropanol to obtain the final product. In one specific embodiment, anhydrous ethanol is used for slurrying.

[0041] Optionally, the preparation methods of compounds E and F are as follows: oxaloyl monoethyl chloride or oxaloyl monomethyl chloride is condensed with glycyl glycinate benzyl toluenesulfonate under alkaline conditions to obtain ethoxyoxaloyl glycyl glycinate benzyl ester or methoxyoxaloyl glycyl glycinate benzyl ester; then hydrogenated to remove the benzyl group to obtain the compounds.

[0042] In the preparation methods of compounds E and F:

[0043] Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran. In one specific embodiment, the selected reaction solvent is dichloromethane.

[0044] Optionally, the alkaline condition is any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. In one specific embodiment, the alkaline condition is triethylamine.

[0045] Optionally, in the preparation method, the molar ratio of oxaloyl chloride monomethyl(ethyl) ester: glycyl glycinate benzyl toluenesulfonate: triethylamine is 1:(1-1.5):(2-3). In one specific embodiment, the molar ratio is 1:1:2.

[0046] Optionally, the extraction solvent used in the preparation method is any one or more of ethyl acetate, methyl acetate, acetone, and butyl acetate. In one specific embodiment, the extraction solvent used is ethyl acetate.

[0047] Optionally, the purification and recrystallization method in the preparation method may use any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile. In one specific embodiment, anhydrous ethanol may be used as the purification and recrystallization method.

[0048] Optionally, the reaction solvent for the debenzylation reaction in the preparation method is any one or more of n-propanol, anhydrous ethanol, anhydrous methanol, ethyl acetate, and tetrahydrofuran. In one specific embodiment, the reaction solvent is anhydrous ethanol.

[0049] Optionally, the catalyst used for debenzylation in the preparation method is any one or more of palladium on carbon, palladium chloride, and palladium oxide. In one specific embodiment, the catalyst used is palladium on carbon.

[0050] Optionally, the purification and recrystallization method used for debenzylation in the preparation method may be any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile. In one specific embodiment, anhydrous ethanol is used as the purification and recrystallization method.

[0051] Optionally, the preparation methods of compound G and compound I are as follows: oxaloyl chloride monomethyl ester or oxaloyl chloride monoethyl ester is obtained by condensation with glycyl glycine methyl ester hydrochloride under alkaline conditions.

[0052] In the preparation methods of compounds G and I:

[0053] Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran. In one specific embodiment, the selected reaction solvent is dichloromethane.

[0054] Optionally, the alkaline condition is any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. In one specific embodiment, the alkaline condition is triethylamine.

[0055] Optionally, in the preparation method, the molar ratio of oxaloyl chloride monomethyl(ethyl) ester: glycyl glycine methyl ester hydrochloride: triethylamine is 1:(1-1.5):(2-3). In one specific embodiment, the molar ratio is 1:1:2.

[0056] Optionally, the extraction solvent used in the preparation method is any one or more of ethanol, ethyl acetate, methyl acetate, acetone, and butyl acetate. In one specific embodiment, the extraction solvent used is a mixture of ethanol and ethyl acetate.

[0057] Optionally, the purification and recrystallization method in the preparation method may use any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile. In one specific embodiment, anhydrous ethanol may be used as the purification and recrystallization method.

[0058] Optionally, the preparation method of compound J is as follows: an intermediate is prepared by condensation of oxalyl chloride monoallyl ester and glycyl glycine ethyl ester hydrochloride under alkaline conditions, followed by hydrogenation.

[0059] In the preparation method of compound J,

[0060] Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran. In one specific embodiment, the selected reaction solvent is dichloromethane.

[0061] Optionally, the alkaline condition is any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. In one specific embodiment, the alkaline condition is triethylamine.

[0062] Optionally, in the preparation method, the molar ratio of oxalyl chloride monoallyl ester: glycyl glycinate ethyl hydrochloride: triethylamine is 1:(1-1.5):(2-3). In one specific embodiment, the molar ratio is 1:1:2.

[0063] Optionally, the extraction solvent used in the preparation method is any one or more of ethyl acetate, methyl acetate, acetone, and butyl acetate. In one specific embodiment, the extraction solvent used is ethyl acetate.

[0064] Optionally, the purification and recrystallization method for the intermediate in the preparation method may use any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile. In one specific embodiment, anhydrous ethanol is used as the purification and recrystallization method for the intermediate.

[0065] Optionally, the hydrogenation reaction solvent in the preparation method is any one or more of n-propanol, anhydrous ethanol, anhydrous methanol, ethyl acetate, and tetrahydrofuran. In one specific embodiment, the hydrogenation reaction solvent is anhydrous ethanol.

[0066] Optionally, the catalyst for the hydrogenation reaction in the preparation method is any one or more of palladium on carbon, palladium chloride, and palladium oxide. In one specific embodiment, the catalyst is palladium on carbon.

[0067] Optionally, the purification and recrystallization method after hydrogenation in the preparation method may use any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile. In one specific embodiment, anhydrous ethanol is used as the purification and recrystallization method after hydrogenation.

[0068] Furthermore, when R1 is -OH and R2 is -OH, the compound is compound A.

[0069] The structural formula of compound A is as follows:

[0070] Optionally, the preparation method of compound A is as follows: using N-oxaloylglycylglycine ethyl ester as raw material, it is subjected to hot hydrolysis, pH adjustment, and filtration in an alkaline solution, and the filtrate is added to a precipitation solvent to induce crystallization.

[0071] In the preparation method of compound A:

[0072] Optionally, the alkaline solution is any one or more aqueous solutions of sodium hydroxide, potassium hydroxide, lithium hydroxide, ammonia, sodium carbonate, potassium carbonate, sodium bicarbonate, and potassium bicarbonate. In one specific embodiment, the alkaline solution is a 1-2 mol / L sodium carbonate solution, at which concentration the reaction is mild and complete.

[0073] Optionally, the hot hydrolysis temperature is 40-60℃.

[0074] Optionally, the precipitation solvent is any one or more of anhydrous ethanol, n-propanol, isopropanol, n-butanol, acetone, and acetonitrile, which are at least six times the volume of the filtrate. In one specific embodiment, the precipitation solvent is anhydrous ethanol.

[0075] Furthermore, when R1 is -OCH2C6H5 and R2 is -OCH2CH3, the compound is compound C.

[0076] The structural formula of compound C is as follows:

[0077] Optionally, the preparation method of compound C is as follows: first, oxaloyl chloride is reacted with benzyl alcohol to obtain oxalate monobenzyl chloride, and then condensed with glycyl glycine ethyl ester hydrochloride under alkaline conditions.

[0078] Secondly, this application provides an application of a compound in evaluating drug quality and safety, employing the following technical solution:

[0079] The application of a compound in evaluating drug quality and safety, the general structural formula of which is shown below:

[0080]

[0081] Wherein, R1 is selected from any one of hydroxyl, methoxy, ethoxy, propoxy, benzyloxy, and -NHCH2CONHCH2COOCH2CH3; R2 is selected from any one of hydroxyl, methoxy, and ethoxy.

[0082] Secondly, this application provides a pharmaceutical composition, which adopts the following technical solution:

[0083] A pharmaceutical composition comprising N-oxoylglycylglycine ethyl ester or a pharmaceutically acceptable salt thereof and a compound with the following general structural formula;

[0084]

[0085] Wherein, R1 is selected from any one of hydroxyl, methoxy, ethoxy, propoxy, benzyloxy, and -NHCH2CONHCH2COOCH2CH3; R2 is selected from any one of hydroxyl, methoxy, and ethoxy.

[0086] Optionally, the content of the compound in the pharmaceutical composition is not higher than 2.0%.

[0087] In summary, this application includes at least one of the following beneficial technical effects:

[0088] This application provides a compound and its preparation method. The preparation method has mild and easy-to-operate reaction conditions, and the product has high yield, high purity, and low cost. It provides a guarantee for the qualitative and quantitative study of N-oxaloylglycylglycine ethyl ester raw materials and related substances, and has significant implementation value and social and economic benefits.

[0089] Compounds A, B, C, D, E, F, G, H, I, and J are impurities generated during the preparation and storage of the drug N-oxaloylglycylglycine ethyl ester. They are used as reference standards in production process quality monitoring and product quality research, and for system suitability, quantitative, and qualitative detection in liquid chromatography. Attached Figure Description

[0090] Figure 1 Here is the HNMR spectrum of compound A.

[0091] Figure 2 This is a partial mass spectrum of compound A (MALDI-TOF MS positive charge mode).

[0092] Figure 3This is the HNMR spectrum of compound B.

[0093] Figure 4 This is a partial mass spectrum of compound B (MALDI-TOF MS, positive charge mode).

[0094] Figure 5 The results are the base peak ion chromatogram and UV chromatogram of compound H.

[0095] Figure 6 The results are the base peak ion chromatogram and UV chromatogram of compound C.

[0096] Figure 7 The results are the base peak ion chromatogram and UV chromatogram of compound D.

[0097] Figure 8 The results are the base peak ion chromatogram and UV chromatogram of compound E.

[0098] Figure 9 The results are the base peak ion chromatogram and UV chromatogram of compound F.

[0099] Figure 10 The results are the base peak ion chromatogram and UV chromatogram of compound G.

[0100] Figure 11 The results are the base peak ion chromatogram and UV chromatogram of compound I.

[0101] Figure 12 The results are the base peak ion chromatogram and UV chromatogram of compound J.

[0102] Figure 13 This is a chromatogram showing the system suitability of drugs and compounds A, B, C, D, and E.

[0103] Figure 14 This is a liquid chromatogram of the drug and compound G.

[0104] Figure 15 The chromatograms are of the drug and compounds H, F, and J. Detailed Implementation

[0105] Before describing the embodiments of this application in detail, it should be understood that the terminology used herein is for the purpose of describing a particular embodiment only. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the term pertains.

[0106] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this application, unless otherwise stated, "multiple" means two or more.

[0107] The endpoints and any values ​​of the ranges disclosed in this application are not limited to the precise ranges or values, and such ranges or values ​​should be understood to include values ​​close to such ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0108] In this application, the terms "comprising" or "including" are open-ended expressions, meaning they include the content specified in this application but do not exclude other aspects.

[0109] To make the objectives, technical solutions, and advantages of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application. The embodiments described below are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0110] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0111] The present application will be further described in detail below with reference to the embodiments and test results.

[0112] Example 1

[0113] This embodiment provides a compound. The compound is compound A, namely (2-[2-(carboxyformamido)acetamido]acetic acid), with the molecular formula C6H8N2O6, molecular weight 204.1, and the structural formula as follows:

[0114]

[0115] The compound is prepared by using N-oxoylglycine ethyl ester as raw material, which is subjected to hot hydrolysis in an alkaline solution, pH adjustment, and filtration. The filtrate is then added to a precipitating solvent to induce crystallization.

[0116] The specific preparation method of this compound is as follows: 10g of N-oxaloylglycylglycine ethyl ester product is added to an Erlenmeyer flask, and 50ml of 2mol / L sodium carbonate solution is added to dissolve it. The mixture is reacted in a water bath at 50℃ for 6 hours. Excess trifluoroacetic acid is slowly added to the filtrate to adjust the pH value to ≤1. The solid is filtered out, and more than 6 times the volume of anhydrous ethanol is added to the filtrate. The mixture is stirred to precipitate crystals, filtered, and dried to obtain the product.

[0117] The reaction process is illustrated in the diagram below:

[0118]

[0119] The product was tested using nuclear magnetic resonance (NMR) and mass spectrometry (MS), and the test results are as follows: Figure 1 and Figure 2 As shown.

[0120] Depend on Figure 1 and Figure 2 The ¹H NMR (DMSO, 400 MHz) values ​​were 8.75–8.68 (m, 1H), 8.23 ​​(t, J = 5.72 Hz, 1H), and 3.86–3.58 (m, 4H); the MS (m / z) value was 205.06 (M+H)+. Therefore, the above compound can be successfully prepared using the method described in this application.

[0121] Example 2

[0122] This embodiment provides a compound.

[0123] The compound is prepared by condensing oxaloyl chloride monoethyl ester or oxaloyl chloride monomethyl ester with glycyl glycinate ethyl ester hydrochloride under alkaline conditions.

[0124] The specific preparation method is as follows:

[0125] 10 g glycyl glycine ethyl ester hydrochloride (0.051 mol) was dissolved in 300 ml dichloromethane by adding 14.2 ml (0.102 mol) triethylamine and stirring. 0.051 mol oxaloyl chloride monoethyl ester (5.7 ml) or oxaloyl chloride monomethyl ester (6.25 g) was added dropwise at 5-15℃. After reacting at low temperature for 2 h, the mixture was raised to room temperature and reacted overnight. The dichloromethane was removed by concentration, and the product was extracted with ethyl acetate / anhydrous ethanol (2 / 1). The extract was concentrated to an appropriate volume, frozen to crystallize, filtered to obtain a solid, and recrystallized with anhydrous ethanol to obtain the final product.

[0126] (1) When prepared using oxaloyl chloride monoethyl ester and glycyl glycinate ethyl ester hydrochloride under alkaline conditions, compound B is obtained, which is ethyl 2-[2-(2-ethoxy-2-oxoacetamido)acetamido], molecular formula: C 10 H 16N2O6, molecular weight: 260.3, structural formula as follows:

[0127]

[0128] The reaction process is illustrated in the diagram below:

[0129]

[0130] The product was tested using nuclear magnetic resonance (NMR) and mass spectrometry (MS), and the test results are as follows: Figure 3 and Figure 4 As shown.

[0131] Depend on Figure 3 and Figure 4 It can be seen that H-NMR (DMSO, 400MHz): 9.03 (t, J = 8Hz, 1H), 8.38 (t, J = 4Hz, 1H), 4.24 (q, J = 8Hz, 2H), 4.09 (q, J = 8Hz, 2H) ), 3.83 (d, J = 4Hz, 2H), 3.77 (d, J = 4Hz, 2H), 1.28 (t, J = 4Hz, 3H), 1.19 (t, J = 4Hz, 3H); MS (m / z): 261.27 (M+H) + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0132] (2) When prepared using monomethyl oxaloyl chloride and glycyl glycinate hydrochloride under alkaline conditions, compound H is obtained, which is ethyl 2-[2-(2-methoxy-2-oxoacetamido)acetamido], molecular formula: C9H 14 N2O6, molecular weight: 246.2, structural formula as follows:

[0133]

[0134] The reaction process is illustrated in the diagram below:

[0135]

[0136] The product was analyzed using LC-MS / MS. Instrument and parameters: Liquid chromatography (Thermo U3000); mobile phase A: 0.1% FA in water, mobile phase B: 0.1% FA in acetonitrile; flow rate: 1 mL / min; wavelength: 214 nm; liquid phase gradient: phase B 0-60%, time 2-20 min; mass spectrometry (Thermo Q Exactive): spray voltage 2.0 kV, first-stage scan resolution 70000, second-stage scan resolution 17500. Detection results are as follows: Figure 5 As shown.

[0137] Depend on Figure 5It can be seen that the relative peak area of ​​the principal component is 100.00%, and the MS (m / z) is 247.09 (M+H). + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0138] Example 3

[0139] This embodiment provides a compound. The compound is compound C, namely benzyloxyoxaloylglycine ethyl ester, with the molecular formula: C2 15 H 18 N2O6, molecular weight: 322.3, structural formula as follows:

[0140]

[0141] The compound is prepared by first reacting oxaloyl chloride with benzyl alcohol to obtain oxalate monobenzyl chloride, and then condensing it with glycyl glycin ethyl ester hydrochloride under alkaline conditions.

[0142] The specific preparation method of this compound is as follows: The ratio of oxalyl chloride to benzyl alcohol is 1.25:1.0. Oxaloyl chloride and 5 times the volume of dichloromethane are added to a three-necked flask. Benzyl alcohol is added dropwise while maintaining the temperature at 5-15℃. After the addition is complete, the temperature is maintained for another 1 hour, and then the reaction is allowed to proceed to room temperature overnight. The solvent and excess oxalyl chloride are removed by concentration. The mixture is then distilled under reduced pressure, and the fraction collected at 73-75℃ / 0.3 mmHg is the monobenzyl oxalate monoacyl chloride. 9.83 g (0.05 mol) glycyl glycine ethyl ester hydrochloride was dissolved in 295 ml of dichloromethane under stirring, and 13.8 ml (0.1 mol) of triethylamine was added dropwise. The temperature was controlled at 5-15℃, and 9.93 g (0.05 mol) monobenzyl oxalate monoacyl chloride was added dropwise. After the addition was completed, the temperature was controlled at 2 h, and then the reaction was raised to room temperature and allowed to proceed overnight. The dichloromethane was removed by concentration, and the mixture was extracted with ethyl acetate under stirring. The extract was washed twice with water, dried, concentrated to an appropriate amount, refrigerated to crystallize, filtered, and dried to obtain the finished product.

[0143] The reaction process is illustrated in the diagram below:

[0144]

[0145] The product was analyzed using LC-MS / MS. Instrument and parameters: Liquid chromatography (Thermo U3000); mobile phase A: 0.1% FA water; mobile phase B: 0.1% FA acetonitrile; flow rate: 0.3 mL / min; wavelength: 214 nm; liquid phase gradient: B phase 2-60%, time 5-20 min; mass spectrometry (Thermo Q Exactive): spray voltage 2.0 kV; primary scan resolution 70000; secondary scan resolution 17500. Detection results are as follows: Figure 6 As shown.

[0146] Depend on Figure 6The relative peak area of ​​the principal component is 97.23%, and the MS (m / z) value is 323.12 (M+H). + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0147] Example 4

[0148] This embodiment provides a compound. The compound is compound D, specifically (2-{2-[({[(2-ethoxy-2-oxoethyl)carbamoyl]methyl}carbamoyl)formamido]acetamido}ethyl acetate), molecular formula: C 14 H 22 N4O8, molecular weight: 374.4, structural formula as follows:

[0149]

[0150] The compound was prepared by condensation of oxaloyl chloride and glycyl glycinate ethyl hydrochloride under alkaline conditions.

[0151] The specific preparation method of this compound is as follows: 10g of glycyl glycine ethyl ester hydrochloride (0.051mol) was stirred with 300ml of dichloromethane, and 28.4ml (0.204mol) of triethylamine was added and stirred until completely dissolved. 2.18ml (0.0255mol) of oxaloyl chloride was added dropwise at 5-15℃. The reaction was carried out at low temperature for 2 hours, and then raised to room temperature and reacted overnight. The dichloromethane was removed by concentration. The solid was first dissolved in ethyl acetate with stirring to dissolve impurities, and then dissolved in hot water at 70℃ with stirring. The insoluble matter was dried to obtain the crude product. The crude product was completely dissolved in 30 times its volume of DMF at 70℃. After decolorization with activated carbon, the filtrate was added to an equal volume of water to induce crystallization. The mixture was filtered, and the filter cake was slurried with anhydrous ethanol. The insoluble matter was filtered out and dried to obtain the product.

[0152] The reaction process is illustrated in the diagram below:

[0153]

[0154] The product was analyzed using LC-MS / MS. Instrument and parameters: Liquid chromatography (Thermo U3000); mobile phase A: 0.1% FA water; mobile phase B: 0.1% FA acetonitrile; flow rate: 1 mL / min; wavelength: 226 nm; column temperature: 35℃; liquid phase gradient: B phase 50-80%, time 2-20 min; mass spectrometry (Thermo Q Exactive): spray voltage 2.0 kV, first-stage scan resolution 70000, second-stage scan resolution 17500. Detection results are as follows: Figure 7 As shown.

[0155] Depend on Figure 7It can be seen that the relative peak area of ​​the principal component is 100%, and the MS (m / z) value is 375.25 (M+H). + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0156] Example 5

[0157] This embodiment provides a compound.

[0158] The compound is prepared by condensing oxaloyl chloride monoethyl ester or oxaloyl chloride monomethyl ester with glycyl glycine benzyl ester p-toluenesulfonate under alkaline conditions to obtain ethoxyoxaloyl glycine benzyl ester or methoxyoxaloyl glycine benzyl ester; then hydrogenating to remove the benzyl group to obtain the compound.

[0159] The specific preparation method is as follows:

[0160] 3.94 g glycyl glycin benzyl ester p-toluenesulfonate (0.01 mol) was stirred with 120 ml dichloromethane. 2.78 ml triethylamine (0.02 mol) was added, and the mixture was completely dissolved. Under an ice-water bath, 1.37 g (0.01 mol) of oxaloyl chloride monoethyl ester or 1.23 g (oxaloyl chloride monomethyl ester) was added dropwise. The mixture was reacted at low temperature for 2 hours, then at room temperature overnight. The product was concentrated to remove dichloromethane, and ethyl acetate was added and stirred to extract the product. The extract was washed twice with water, dried over anhydrous sodium sulfate, evaporated to dryness, and recrystallized from anhydrous ethanol to obtain a white solid. Deprotection: The product was dissolved in anhydrous ethanol, and 0.1 times the volume of 10% palladium on carbon was added. After several gas changes, hydrogen gas was passed through, and the mixture was reacted overnight at room temperature. The palladium on carbon was filtered off, the solvent was concentrated to a suitable volume, and crystals were precipitated to obtain the final product.

[0161] (1) When oxaloyl chloride monoethyl ester and glycyl glycine benzyl ester p-toluenesulfonate are condensed under alkaline conditions to obtain ethoxyoxaloyl glycine benzyl ester; and then hydrogenated to remove the benzyl group to obtain compound E, which is (2-[2-(2-ethoxy-2-oxoacetamido)acetamido]acetic acid, molecular formula: C8H 12 N₂O₆, molecular weight: 232.2, structural formula as follows:

[0162]

[0163] The reaction process is illustrated in the diagram below:

[0164]

[0165] The product was analyzed using LC-MS / MS. Instrument and parameters: Liquid chromatography (Thermo U3000); mobile phase A: 0.1% FA water; mobile phase B: 0.1% FA acetonitrile; flow rate: 0.3 mL / min; wavelength: 214 nm; liquid phase gradient: B phase 2-60%, time 5-20 min; mass spectrometry (Thermo Q Exactive): spray voltage 2.0 kV; primary scan resolution 70000; secondary scan resolution 17500. Detection results are as follows: Figure 8 As shown.

[0166] Depend on Figure 8 The relative peak area of ​​the principal component is 99.66%, and the MS (m / z) value is 233.08 (M+H). + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0167] (2) When oxaloyl chloride monomethyl ester and glycyl glycinate benzyl ester p-toluenesulfonate are condensed under alkaline conditions to obtain methoxyoxaloyl glycyl glycinate benzyl ester; and then hydrogenated to remove the benzyl group, compound F is obtained, which is (2-[2-(2-methoxy-2-oxoacetamido)acetamido]acetic acid, molecular formula: C7H 10 N2O6, molecular weight: 218.2, structural formula as follows:

[0168]

[0169] The reaction process is illustrated in the diagram below:

[0170]

[0171] The product was analyzed using LC-MS / MS. Instrument and parameters: Liquid chromatography (Thermo U3000); mobile phase A: 0.1% FA water; mobile phase B: 0.1% FA acetonitrile; flow rate: 0.3 mL / min; wavelength: 214 nm; liquid phase gradient: B phase 2-60%, time 5-20 min; mass spectrometry (Thermo Q Exactive): spray voltage 2.0 kV; primary scan resolution 70000; secondary scan resolution 17500. Detection results are as follows: Figure 9 As shown.

[0172] Depend on Figure 9 The relative peak area of ​​the principal component is 99.50%, and the MS (m / z) value is 219.06 (M+H). + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0173] Example 6

[0174] This embodiment provides a compound.

[0175] The compound is prepared by condensing oxaloyl chloride monomethyl ester or oxaloyl chloride monoethyl ester with glycyl glycine methyl ester hydrochloride under alkaline conditions.

[0176] The specific preparation method is as follows:

[0177] 9.13 g glycyl glycine methyl ester hydrochloride (0.05 mol) was dissolved in 300 ml of dichloromethane with 13.9 ml (0.1 mol) triethylamine and stirred until completely dissolved. 0.05 mol oxaloyl chloride monomethyl ester (6.13 g) or oxaloyl chloride monoethyl ester (6.83 g) was added dropwise at 5-15℃. After reacting at low temperature for 2 h, the mixture was raised to room temperature and reacted overnight. The dichloromethane was removed by concentration, and the product was extracted with ethyl acetate / anhydrous ethanol (1 / 1). The extract was concentrated to an appropriate volume, frozen to crystallize, filtered to obtain a solid, and recrystallized with anhydrous ethanol to obtain the final product.

[0178] (1) When oxaloyl chloride monomethyl ester and glycyl glycine methyl ester hydrochloride are condensed under alkaline conditions, compound G is obtained, which is methyl 2-[2-(2-methoxy-2-oxoacetamido)acetamido]acetate, with the molecular formula: C8H 12 N₂O₆, molecular weight: 232.2, structural formula as follows:

[0179]

[0180] The reaction process is illustrated in the diagram below:

[0181]

[0182] The product was analyzed using LC-MS / MS. Instrument and parameters: Liquid chromatography (Thermo U3000); mobile phase A: 0.1% FA in water, mobile phase B: 0.1% FA in acetonitrile; flow rate: 1 mL / min; wavelength: 214 nm; liquid phase gradient: phase B 0-60%, time 2-20 min; mass spectrometry (Thermo Q Exactive): spray voltage 2.0 kV, first-stage scan resolution 70000, second-stage scan resolution 17500. Detection results are as follows: Figure 10 As shown.

[0183] Depend on Figure 10 It can be seen that the relative peak area of ​​the principal component is 100.00%, and the MS (m / z) is 233.06 (M+H). + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0184] (2) When oxaloyl chloride monoethyl ester and glycyl glycine methyl ester hydrochloride are condensed under alkaline conditions, compound I is obtained, which is methyl 2-[2-(2-ethoxy-2-oxoacetamido)acetamido]acetate, with the molecular formula: C9H 14N2O6, molecular weight: 246.2, structural formula as follows:

[0185]

[0186] The reaction process is illustrated in the diagram below:

[0187]

[0188] The product was analyzed using LC-MS / MS. Instrument and parameters: Liquid chromatography (Thermo U3000); mobile phase A: 0.1% FA in water, mobile phase B: 0.1% FA in acetonitrile; flow rate: 1 mL / min; wavelength: 214 nm; liquid phase gradient: phase B 0-60%, time 2-20 min; mass spectrometry (Thermo Q Exactive): spray voltage 2.0 kV, first-stage scan resolution 70000, second-stage scan resolution 17500. Detection results are as follows: Figure 11 As shown.

[0189] Depend on Figure 11 The relative peak area of ​​the principal component is 90.28%, and the MS (m / z) value is 247.07 (M+H). + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0190] Example 7

[0191] This embodiment provides a compound. The compound is compound J, specifically ethyl acetate (2-[2-(2-n-propoxy-2-oxoacetamido)acetamido], with the molecular formula: C2. 11 H 18 N2O6, molecular weight: 274.3, structural formula as follows:

[0192]

[0193] The compound is prepared by condensing oxaloyl chloride monoallyl ester and glycyl glycinate ethyl ester hydrochloride under alkaline conditions to obtain an intermediate, followed by hydrogenation.

[0194] The specific preparation method of this compound is as follows:

[0195] 10g glycyl glycine ethyl ester hydrochloride (0.051mol) was dissolved in 300ml dichloromethane with 14.2ml (0.102mol) triethylamine by stirring. 7.58g (0.051mol) oxalyl chloride monoallyl ester was added dropwise at 5-15℃. After reacting at low temperature for 2 hours, the mixture was allowed to react overnight at room temperature. The product was concentrated to remove dichloromethane, and ethyl acetate was added to extract the product. The extract was washed twice with water, dried, concentrated to an appropriate volume, refrigerated to crystallize, filtered, dried, and recrystallized with anhydrous ethanol to obtain the final product.

[0196] Dissolve the finished product in anhydrous ethanol, add 0.1 times the volume of 10% palladium on carbon, purge several times, and then pass hydrogen gas through the solution to react overnight at room temperature. Filter off the palladium on carbon, concentrate the solvent to a suitable volume, cool to allow crystals to precipitate, and filter to obtain the finished product. Recrystallize from anhydrous ethanol.

[0197] The reaction process is illustrated in the diagram below:

[0198]

[0199] The product was analyzed using LC-MS / MS. Instrument and parameters: Liquid chromatography (Thermo U3000); mobile phase A: 0.1% FA in water, mobile phase B: 0.1% FA in acetonitrile; flow rate: 1 mL / min; wavelength: 214 nm; liquid phase gradient: phase B 0-60%, time 2-20 min; mass spectrometry (Thermo Q Exactive): spray voltage 2.0 kV, first-stage scan resolution 70000, second-stage scan resolution 17500. Detection results are as follows: Figure 12 As shown.

[0200] Depend on Figure 12 The relative peak area of ​​the principal component is 90.91%, and the MS (m / z) value is 275.10 (M+H). + Therefore, the above-mentioned compound can be successfully prepared using the preparation method of this application.

[0201] Example 8

[0202] This embodiment provides an example of using a compound to evaluate the quality and safety of a drug. The drug is N-oxaloylglycine ethyl ester.

[0203] The specific details are as follows: The drug N-oxaloylglycylglycine ethyl ester is prepared in ethanol. The main impurities are compounds A, B, C, D, and E. Compound A is a drug degradation impurity; compounds B (from the reaction of the product with ethanol), C, and D are process impurities; and compound E is a degradation impurity of compound B. These impurities are unavoidable in pharmaceuticals, and their content must be strictly limited to ensure drug quality and safety.

[0204] Weigh appropriate amounts of N-oxaloylglycylglycine ethyl ester and compounds A, B, C, D, and E to dissolve them, preparing a system suitability solution. Liquid chromatography was performed using a Thermo U3000 column packed with C18 material. Mobile phase A consisted of 0.25 mol / L formic acid solution, and mobile phase B consisted of acetonitrile at a flow rate of 1.0 mL / min. The detection wavelength was 214 nm. A gradient was applied to phase B from 0-40% for 0-40 min, followed by phase B from 40-0% for 40-60 min. The detection results are as follows: Figure 13 As shown.

[0205] Depend on Figure 13 It can be seen that, under suitable liquid chromatography conditions, the drug and compounds A, B, C, D, and E meet the system suitability requirements in terms of resolution and theoretical plate number, and can be used as system suitability references to confirm the effectiveness and stability of the liquid chromatography analysis system. Furthermore, the locations of compounds A, B, C, D, and E in the liquid chromatography were determined, and their retention times were compared with those of impurities in the test sample to identify the types and quantitative ranges of impurities. This further confirms whether the drug meets the proposed quality standards.

[0206] The drug N-oxoylglycylglycine ethyl ester was prepared in methanol. The main impurities were compounds A, H, C, D, F, and G. Compounds H (the product reacts with the solvent methanol), C and D were process impurities, compound F was a degradation impurity of compound H, and compound G was a transesterification product in methanol.

[0207] The drug N-oxaloylglycylglycine ethyl ester is prepared in the solvent n-propanol. The largest impurity is compound J (the product reacts with the solvent n-propanol). Different impurities are generated in different processes, and it is necessary to identify these impurities and establish reasonable limit standards.

[0208] Under the same liquid phase conditions, an appropriate amount of N-oxoylglycylglycine ethyl ester and compound G were dissolved, and the injection detection results were as follows: Figure 14 As shown. Dissolve appropriate amounts of N-oxaloylglycylglycine ethyl ester with compounds H, F, and J. The injection and detection results are as follows. Figure 15 As shown.

[0209] Depend on Figure 14 and 15 It can be seen that compounds H, F, G, and J were located in liquid chromatography, and their relative retention times were obtained. By comparing these times with the retention times of impurities in the test sample, the types and quantitative ranges of the impurities can be determined. This confirms that the drug meets the proposed quality standards.

[0210] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0211] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. A compound, characterized in that, The general structural formula of the compound is shown below: Wherein, R1 is selected from any one of hydroxyl, methoxy, ethoxy, propoxy, benzyloxy, and -NHCH2CONHCH2COOCH2CH3; R2 is selected from any one of hydroxyl, methoxy, and ethoxy; and R1 and R2 are not both hydroxyl, nor are they both benzyloxy and ethoxy.

2. The compound according to claim 1, characterized in that, The compounds include compounds B, D, E, F, G, H, I, and J; The structural formulas are shown below: Compound B Compound D Compound E Compound F Compound G Compound H Compound I Compound J 3. The compound according to claim 2, characterized in that, The specific preparation methods of compounds B and H are as follows: they are obtained by condensation of oxaloyl chloride monoethyl ester or oxaloyl chloride monomethyl ester with glycine ethyl ester hydrochloride under alkaline conditions; Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran; Optionally, the alkaline conditions are any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. Optionally, in the preparation method, the molar ratio of oxaloyl chloride monomethyl(ethyl) ester: glycyl glycine ethyl ester hydrochloride: triethylamine is 1:(1-1.5):(2-3). Optionally, the extraction solvent used in the preparation method is any one or more of ethanol, ethyl acetate, methyl acetate, acetone, and butyl acetate; Optionally, the purification and recrystallization method in the preparation method may be any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile.

4. The compound according to claim 2, characterized in that, The specific method for preparing compound D is as follows: It is obtained by condensation of oxaloyl chloride and glycyl glycine ethyl ester hydrochloride under alkaline conditions; Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran; Optionally, the alkaline conditions are any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. Optionally, the preliminary purification method in the preparation method is to dissolve the ester-soluble compound with ethyl acetate, methyl acetate, acetone, and butyl acetate, and dissolve the water-soluble compound with water at 60-90°C, with the remaining solid being the crude product; Optionally, the further purification method in the preparation method involves thermal dissolution of DMF or DMSO, decolorization and filtration with activated carbon, addition to water to precipitate the solid, and then slurrying with anhydrous ethanol, methanol or isopropanol to obtain the final product.

5. The compound according to claim 2, characterized in that, The specific preparation methods of compounds E and F are as follows: oxaloyl chloride monoethyl ester or oxaloyl chloride monomethyl ester is condensed with glycyl glycine benzyl ester p-toluenesulfonate under alkaline conditions to obtain ethoxyoxaloyl glycine benzyl ester or methoxyoxaloyl glycine benzyl ester; then hydrogenated to remove the benzyl group to obtain the compounds. Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran; Optionally, the alkaline conditions are any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. Optionally, in the preparation method, the molar ratio of oxaloyl chloride monomethyl(ethyl) ester: glycyl glycinate benzyl ester p-toluenesulfonate: triethylamine is 1:(1-1.5):(2-3). Optionally, the extraction solvent used in the preparation method is any one or more of ethyl acetate, methyl acetate, acetone, and butyl acetate; Optionally, the purification and recrystallization method in the preparation method may use any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile. Optionally, the reaction solvent for debenzyl ester protection in the preparation method is any one or more of n-propanol, anhydrous ethanol, anhydrous methanol, ethyl acetate, and tetrahydrofuran; Optionally, the catalyst used for debenzyl ester protection in the preparation method is any one or more of palladium on carbon, palladium chloride, and palladium oxide; Optionally, the purification and recrystallization method used for debenzylidene protection in the preparation method may be any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile.

6. The compound according to claim 2, characterized in that, The specific preparation methods of compound G and compound I are as follows: oxaloyl chloride monomethyl ester or oxaloyl chloride monoethyl ester is obtained by condensation with glycyl glycine methyl ester hydrochloride under alkaline conditions; Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran; Optionally, the alkaline conditions are any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. Optionally, in the preparation method, the molar ratio of oxaloyl chloride monomethyl(ethyl) ester: glycyl glycine methyl ester hydrochloride: triethylamine is 1:(1-1.5):(2-3). Optionally, the extraction solvent used in the preparation method is any one or more of ethyl acetate, methyl acetate, acetone, and butyl acetate; Optionally, the purification and recrystallization method in the preparation method may be any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile.

7. The compound according to claim 2, characterized in that, The specific preparation method of compound J is as follows: an intermediate is prepared by condensation of oxalyl chloride monoallyl ester and glycyl glycine ethyl ester hydrochloride under alkaline conditions, followed by hydrogenation to obtain the compound J. Optionally, the reaction solvent selected in the preparation method is any one or more of dichloromethane, trichloromethane, dichloroethane, and tetrahydrofuran; Optionally, the alkaline conditions are any one or more of triethylamine, diisopropylethylamine, potassium bicarbonate, sodium bicarbonate, ammonia, sodium carbonate, and potassium carbonate. Optionally, in the preparation method, the molar ratio of oxalyl chloride monoallyl ester: glycyl glycine ethyl ester hydrochloride: triethylamine is 1:(1-1.5):(2-3). Optionally, the extraction solvent used in the preparation method is any one or more of ethyl acetate, methyl acetate, acetone, and butyl acetate; Optionally, the purification and recrystallization method for the intermediate in the preparation method may use any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile. Optionally, the hydrogenation reaction solvent in the preparation method is any one or more of n-propanol, anhydrous ethanol, anhydrous methanol, ethyl acetate, and tetrahydrofuran; Optionally, the catalyst for the hydrogenation reaction in the preparation method is any one or more of palladium on carbon, palladium chloride, and palladium oxide; Optionally, the purification and recrystallization method after hydrogenation reaction in the preparation method may be any one or more of anhydrous ethanol, methanol, isopropanol, n-propanol, ethyl acetate, and acetonitrile.

8. The application of a compound in evaluating drug quality and safety, characterized in that, The general structural formula of the compound is shown below: Wherein, R1 is selected from any one of hydroxyl, methoxy, ethoxy, propoxy, benzyloxy, and -NHCH2CONHCH2COOCH2CH3; R2 is selected from any one of hydroxyl, methoxy, and ethoxy.

9. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises N-oxoylglycine ethyl ester or a pharmaceutically acceptable salt thereof, and compounds with the following general structural formula; Wherein, R1 is selected from any one of hydroxyl, methoxy, ethoxy, propoxy, benzyloxy, and -NHCH2CONHCH2COOCH2CH3; R2 is selected from any one of hydroxyl, methoxy, and ethoxy.

10. The pharmaceutical composition according to claim 9, characterized in that, The content of the compound in the pharmaceutical composition is not higher than 2.0%.

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