Study On The Pilot Process(Including Wastes Treatment) And The Product Quality Of Aditoprim API

Aditoprim is a novel benzylamine pyrimidine antibacterial synergist with higher safety and better antibacterial effect than other drugs in this class,which is developed as I class new veterinary pharmaceuticals.Up to now,it has not been listed in any county.Pharmacological research on Aditoprim is an important foundation to ensure its safety,effectiveness and quality control.Based on the previous process research and quality research,according to the guiding principles of veterinary drug preparation and quality research,this study carried out pilot-scale process research and GMP pilot-scale production of Aditoprim,and established quality standards on the basis of total quality research on pilot-scale products.Meanwhile,the wastes treatment process in the production process of Aditoprim was studied.Through the above research,we can provide qualified drugs for the formulation development and later clinical experiment of Aditoprim.1. Process study of Aditoprim APIAditoprim API synthesis process is based on p-dimethylaminobenzaldehyde as API,after reduction,bromo-,methoxy,cyanide reduction-reduction amine,condensation,cyclization reaction and six-step reactions to obtain the target product.The synthetic process has been completed in our lab,but in condensation,cyclization reaction,there are many impurities and low yield in these two steps.In order to better adapt to the pilot production,improve the yield and ensure the quality of products iare important.This study aimed at the optimization of these two steps.The two-step reaction was optimized by orthogonal test to determine the condensation with reference to TMP and Bromoprim.Processes:acrylonitrile:sodium methoxide:ADP_4,The feeding ratio is 1.1:2.5:1.0,the reaction temperature is 35℃,the reaction time is 6.5 h;the cyclization process is:guanidine nitrate:sodium methoxide:ADP_5,The reaction temperature was 120℃and the reaction time was 8 h.And the optimized two-step reaction yield increased to75.9%～82.8%and 84.4%～87.5%respectively.The refining process of Aditoprim is recrystallization,From the crystalline solvent to the refining process was optimized by solvent usage and crystallization temperature.Results:60%methanol/water was used as solvent,solvent usage 1.2 g/m L,reflux 1 h,filtration while hot,-10℃static crystallization,the obtained crystallization after filtration is obtained at 60℃vacuum drying for 12 h.And the yield was 96.1%～98.4%,white powder crystal,content was 98.8%～99.3%.The relevant substances are inspected in accordance with the quality standard.The complete process of Aditoprim synthesis was verified after the process optimization.Firstly,the production was gradually enlarged from 200g to 4000g in the laboratory.The results showed that the appearance color of each batch of products was white powder crystal,the content was between 98.8%～99.3%,and the inspection of related substances met the quality standard;Then according to the new drug approval requirements,three consecutive batches of Aditoprim API carry on GMP pilot test in September 2018 in Jingshan Ruisheng Pharmaceutical Co Ltd,obtained Aditoprim API28.27 kg,26.17 kg,26.95 kg,batch numbers of 20180915,20180920,20180925respectively.2. Quality study of pilot product of Aditoprim APIOur group has carried on the preliminary quality research to Aditoprim in the early stage,has established the quality standard draft.This research mainly carries on the inspection to the process verification product,especially the pilot test product according to the quality standard draft,focuses on the inspection appearance,the related substance and the content.By analyzing the related substances of different batches of laboratory products and GMP pilot-scale products,the impurity spectrum is further analyzed,and the inspection method of related substances is determined;The content of batch products is determined by HPLC and non-water titration respectively.The results showed that there were 3 related substances in different batches of the Aditoprim was optimized process,the retention time was 8.4 min,16.0 min,27.2 min,which were same with the previous quality study,and the impurity content was between 0.14%～0.22%,0.15%～0.25%,0.15%and 0.20%respectively.The content of three batches of GMP pilot-scale products was determined by non-aqueous titration,which was 99.05%,98.76%and 99.28%respectively,and determined by non-aqueous titration method,which was 99.13%,98.82%and 99.31%respectively.The melting point,dry weight loss and incandescent residue of three batches of GMP pilot-scale products were examined according to the draft quality standard.The results showed that the melting points were 215.4℃～216.7℃,217.3℃～218.4℃and215.4℃～216.7℃respectively.The dry weight loss was less than 1.0%and the incandescent residue was less than 0.3%.On the basis of the above research on the quality of pilot-scale Aditoprim API,the quality standard of Aditoprim API was further formulated.3. Study on the stability of Aditoprim APIAccording to the Technical Guidelines for the Study of the Stability of Veterinary Chemical Drugs,the influence factor test,accelerated test GMP long-term stability test were carried out on the pilot-scale product of Aditoprim API.Using 20180915 batch as sample,under the conditions of 60℃temperature（high temperature）,90±5%relative humidity（high humidity）,450±500 lx of light intensity（light）for 10 days,On 5 days and10 days,detect the influencing factors according to the stability research project.The results show that under high temperature,0.12%increase in related substances,the content decreased by 0.13%.The related substances increased by 0.49%under strong light,The content decreased by 0.40%.The related substances increased by 0.36%under high humidity,water increased by 0.42%,content decreased by 0.85%.The experimental results of the influencing factors proved that the high temperature of the drug was relatively.The content decreased obviously under light and high humidity.Therefore,it should be kept dry and avoid light.Accelerated test using three batches GMP pilot API,As pertentative packing（the outer packing is molded into small drums,built-in foil packaging）,6 months at a temperature of 40±2℃and a relative humidity of 75±5%.At the end of 1,2,3 and 6 months,the stability test items were investigated.The results of three batches of samples were compared with those of 0 month,no change in color,no change in substance content or type,the contents decreased by 0.18%,0.12%and 0.15%respectively.Three batches of long-term test according to tentative packing（the outer packing used is moulded small barrel,built-in foil packaging）,under conditions of25±2℃,60%relative humidity±10%,In 0,3,6,9,12,24 months,sampling and stability tests,12 months long-term stability study has been conducted,compared with the initial results of 0 month,the results of three batches of samples in each test index were compared.No change in color,no change in substance content or type,the content decreased by 0.11%,0.09%and 0.12%respectively.The long-term stability investigation continues.4.Study on the wastes treatment of Aditoprim APIThe wastes in the production process of Aditoprim API are mainly liquid wastes and residue wastes.The main solvents used include toluene,methanol,formic acid and ethylene glycolmonomethyl ether.In which methanol and ethylene glycol monomethyl ether due to water content requirements,it is necessary to separate and remove water by distillation and azeotropic method,the rest of the solvent is applied directly.The yield of each solvent was investigated after 3 times of repeated application.The results showed that the application of solvent had no obvious effect on the process.Each step extraction water washing liquid,solid product water washing liquid wastewater centralized treatment,the p H value of three pilot-scale liquid wastes treatment in the laboratory was10.71～11.23,the chroma was 61～73,the suspended substance was 16.6～25.9,the five-day biochemical oxygen demand（BOD₅）was 219,the chemical oxygen demand（CODcr）was 3906～4512,the total organic nitrogen（TON）was 39～42,TOC was176～186,After four steps of evaporation,desalination,p H condensation,Fenton oxidation and microbial treatment,The p H value of three batches of liquid wastes was6.97～7.68,chroma was 7,suspended substance was 0.0,5 days biochemical oxygen demand（BOD₅）was 11,chemical oxygen demand（CODcr）was 11～14,total organic nitrogen（TON）was 6～7,total organic carbon（TOC）was 10～12,after four-step treatment of three batches of wastewater,each index is in accordance with the discharge standard（GB 21904-2008）of water pollutants in chemical synthetic pharmaceutical industry.Direct emissions;the residue waste includes the third step containing copper,sodium bromide,activated carbon filter cake and the fourth step deactivating Renny nickel.The waste residue in the third step can be dissolved and refined to obtain different grades of copper and sodium bromide.The raney-nickel was activated by immersion in 30%sodium hydroxide solution for 24 h.,the activity was determined 51 m LH₂/g-Raney-Ni,above quality standards（42 m LH₂/g-Raney-Ni）,can be applied.This process has achieved full recovery and utilization of residue waste and solvent,reached the goal of economic and effective treatment,achieved full and effective treatment of liquid waste,reached the principle of green environmental protection treatment,and has important guiding significance and application value for the treatment of wastes in the industrial production of Aditoprim API,and provided reference for the treatment technology of wastes in the same industry.This study studied the pilot-scale production process and GMP pilot-scale production of Aditoprim API by the first time,carried out quality research on pilot-scale products,and carried out stability tests on three batches of GMP pilot-scale products to further improve the quality standard of Aditoprim API;studied the waste treatment process to ensure the economic safety and green environmental protection of Aditoprim API production.Through the above research,the pharmaceutical research of Aditoprim API has been improved,which provides qualified API for the formulation production and clinical research of Aditoprim,and lay a solid foundation for the new drug creation and industrialization of Aditoprim.