Study On Synthesis And Quality Control Of Broad-Spectrum Antiviral Drug EIDD-2801

The target drug of this study,EIDD-2801,is a chemically synthesized antiviral innovative drug that has not been listed at home or abroad.It is an isobutyl precursor of the ribonucleotide analogue EIDD-1931（N4-hydroxycytidine,NHC）.It is a broad-spectrum antiviral drug with inhibitory effect on influenza virus,Ebola,equine encephalitis virus,hepatitis C virus,coronavirus and other viruses.It has the advantages of high metabolic stability,high bioavailability,high lung distribution rate,broad spectrum and low toxicity.EIDD-2801 was approved by FDA on April 17,20,and entered into clinical trials urgently.This project is divided into two parts.Firstly,EIDD-2801 API is synthesized.In the second part,the study on quality of EIDD-2801,which was synthesized independently,including the physical and chemical properties of the drug,the determination of content and the inspection of related substances,and the stability of the drug was studied.The first part of this project:synthesis of drug EIDD-2801.In this chapter,the synthetic process was explored based on the patent route of the original research company.First,using uridine as raw material,acetone was used to protect the hydroxyl group of sugar ring,and then using isobutyric anhydride to replace methyl alcohol to produce esterification compounds.Then nucleophilic substitution occurs in the presence of phosphorus trichloride,the carbonyl group in the six-member ring was replaced with triazolium,and then in an aqueous solution of hydroxylamine,triazole was converted to hydroxylamine,and then the hydroxyl group in the protective sugar ring was reduced in the action of trifluoroacetic acid and an appropriate amount of water to obtain the crude product of EIDD-2801.After purification,EIDD-2801 was obtained with a yield of 42.42%in terms of uridine and a chromatographic purity of 99.81%in the area normalization method.The structure was characterized by UV,IR,¹H-NMR,¹³C-NMR,HPLC and HPLC-MS,and the results show that the structure is consistent with EIDD-2801 molecular formula.The second part of this topic:quality research of EIDD-2801.In this chapter,the character and physical and chemical properties of the drug of the self-made EIDD-2801 were studied by referring to the fourth part of Chinese Pharmacopoeia（2020 edition）.The general impurity of this product including moisture,dry weight loss,incandescent residue,residual solvent and so on were systematically studied,the determination method of residual solvent was established,and the methodology was verified.According to the structure and physical and chemical properties of EIDD-2801,the related substances detection and the content determination method of EIDD-2801 were established.The chromatographic conditions were as follows:Phenomenex Gemini 5μm C₁₈110A（250×4.6mm）;detection wavelength:235 nm;flow rate:1 m L/min;solvent:30%methanol in water;mobile phase:methanol-water（30:70,v/v）;column temperature:30℃;sample size:10μl;the injection concentration was 0.2mg/m L,and the methodology was verified.According to the stability guidelines of Chinese Pharmacopoeia（2020 edition）,the stability of EIEE-2801 API was studied,and the structure of the main impurities was studied.The results show that it is confirmed that the product have the same structural formula and molecular weight as the target compound EIDD-2801 after structural study.It is white powder crystal,odorless,almost without moisture absorption,soluble in dimethyl sulphoxide,dissolved in methanol,slightly dissolved in water,and slightly dissolved in acetonitrile.The specific rotation is-18.24°,and the melting point is 156～158℃.All the residual solvents are lower than the limit requirements of Chinese Pharmacopeia（2020 Edition）.The main impurity is EIDD-1931（N4-hydroxycytidine,NHC）,which is the hydrolyzed product generated by the ester bond fracture of EIDD-2801.The content determination results of three batches of API show that the content is 102.77%,99.56%and 99.37%respectively.Within three months,stability test results show that the content of this product is stable,no obvious degradation phenomenon.In conclusion,the results show that the established determination methods are applicable to the quality control of EIDD-2801,and provide a good research basis for the establishment of quality standard and process optimization of EIDD-2801.In the quality research,the same conditions and methods are used to simultaneously determine the relevant substances and content of this product,which improves the efficiency of quality control and is beneficial to cost saving and environmental protection.