Study On The Synthesis Process And Related Substances Of Ticagrelor

Ticagrelor,the?1S,2S,3R,5S?-3-[7-[?1R,2S?-2-?3,4-Difluorophenyl?cyclopropylamino]-5-?propylsulfanyl?-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl]-5-?2-hydroxyethoxy?cyclopentane-1,2-diol,is the first small molecule anticoagulant drug developed by astrazeneca in the United Kingdom with reversible binding,direct effect and oral administration.Clinical studies have shown that ticagrelor has the advantages of quickly exert the effect,small individual difference and reduced bleeding risk.It is widely used at home and abroad.The synthetic route of ticagrelor is complex and its chemical structure contains six chiral centers,so there may be several organic impurities in the finished product,including non-isomer impurities and isomer impurities.Firstly,the synthetic route of ticagrelor was determined and the synthesis process suitable for industrial production was established.On this basis,the possible organic impurities in the finished product were analyzed and HPLC methods for the determination of related substances and isomers were established.This study provides an important basis for developing more comprehensive quality control measures of ticagrelor.It also provides theoretical basis and means for evaluating the drug quality of ticagrelor.PART 1 Determine the synthetic route and process of ticagrelorObjective:To determine the synthetic route and process for industrial production of ticagrelor.It provides an example for the study of impurities.Methods:By quering literature and analyzing research,we determined the synthetic route adopted by most manufacturers and established the synthesis process suitable for industrial production.The process route was used to simulate the synthesis of the target product.We also confirmed the structure of the product.Results:Three key intermediates-pyrimidine ring,five-membered ring and three-membered ring were used as starting materials.Ticagrelor was synthesized by four steps:nucleophilic substitution,diazotization loop,nucleophilic substitution and deprotection.It was detected by IR,UV,MS and NMR and the chemical structure of the target product was determined.Conclutions:The process is simple and feasible,suitable for enlarged production.Qualified target products can be prepared and used for impurity analysis and validation.PART 2 Study on the related substances of ticagrelor1.Determination of the related substances of ticagrelor by High Performance Liquid Chromatography.Objective:To establish a HPLC method for the determination of various related substances in ticagrelor.Methods:Chromatographic condition consisted of a reversed-phase analytical column of YMC-Pack ODS-AQ C-18?150×4.6 mm,3?m?.The gradient elution consisted of 10 mmol/L sodium dihydrogen phosphate buffer and acetonitrile.The detection wavelength is 242 nm and the column temperature is 40?.The flow rate is 1.0 m L/min and sample quantity is 10?l.Results:Under the established chromatographic conditions,the impurities can be completely separated from each other and from ticagrelor.This indicates that the specificity of the method is good.The linear relationship between the impurities and ticagrelor was good in the studied concentration range.The regression coefficient was greater than 0.999.Precision,repeatability,accuracy,durability,detection limit and quantification limit all meet the requirements.The content of each impurity in three batches of ticagrelor samples was less than 0.05%,the sum of the impurities is less than 0.3%.The impurity profile is almost consistent with the original product.Conclutions:The method can control all possible non-isomer impurities.The method has good specificity,reproducibility,sensitivity and accuracy.It is applicable to the quality control of the related substances of ticagrelor.2.Determination of the isomers of ticagrelor by High Performance Liquid Chromatography.Objective:To establish a HPLC method for the determination of the isomers of ticagrelor.Methods:We used a chiral chromatographic column that filled with three-?3,5-dimethylcarbamate?amylose bonded silica gel.The mobile phase is n-hexane-anhydrous ethanol-trifluoroacetic acid?85:15:0.5?and the detection wavelength is 255 nm.The flow rate is 1.0 m L/min and the column temperature is 40?.Results:Under the established chromatographic conditions,the isomers have good specificity and separation from ticagrelor.In the corresponding linear range,the linear relationship is good.The detection limits of ticagrelor,Y_1,Y_2,Y₂?,Y_3,Y₃?,Y_4,Y₄?are 0.0032%,0.0057%,0.0030%,0.0034%,0.0033%,0.0034%,0.0032%and 0.0053%respectively;Limit of quantitations are 0.013%,0.023%,0.012%,0.014%,0.013%,0.014%,0.013%and 0.021%respectively.These all can meet the detection sensitivity requirements.Repeatability,precision and accuracy meet the requirements.The chromatographic conditions had no significant effect on the separation degree between peaks.The method has good durability.Conclutions:The method can effectively separate the isomer impurities of ticagrelor.The method has good specificity,reproducibility,sensitivity and accuracy.It is applicable to the quality control of the isomers of ticagrelor.