| Fibrinogen is a macromolecular glycoprotein compound composed of a pair of polypeptide chains secreted by liver and vascular endothelial cells in plasma,which is an important blood coagulation factor in plasma.It has hemostasis,blocking and adhesion effect by combining with thrombin.In addition,fibrinogen is often used as a military reserve hemostatic drug and is included in the national emergency.However,due to the high cost of human plasma fibrinogen and the few blood product production units in the country,the fibrinogen preparation has been in short supply in the clinical situation for a long time,and the clinical requirements of raw material safety and formulation performance need to be further improved But,As a new type of animal-derived fibrinogen,it has received extensive attention.Because it not only prevents the danger of virus transmission,but also greatly reduces production costs.This research aims to use a new fully enclosed aseptic technique was used to collect plasma from healthy pigs,and the obtained pig plasma was used to develop fibrinogen preparations,and appropriate protective agent formulations and freeze-drying processes were selected to improve the performance of fibrinogen preparations,quality and cost savings.Research content: 1?The protective agents for pig fibrinogen preparations were studied andscreened.Four kinds of protective agents with great influence on theperformance of the preparations were selected by literature,patent andsingle factor experiments.The response surface was optimized byBox-Behnken experimental design.2?Using electric resistance measurement method to meaurse the eutecticpoint and eutectic point of the formulation,and carrying out thepre-freezing temperature,time,annealing and other,using the apparentanalyzer such as Micro-CT technique,scanning electron microscope?SEM?and circular dichroism to explore the microscopic structuralproperties and physicochemical properties of the porcine fibrinogenfreeze-dried product during lyophilization.Optimizing and adjust thefreeze-drying process of porcine fibrinogen preparation to improveproduct quality.3?Using small test amplification to make three batches of products,andcarrying out the stability study and accelerated stability experimentalstudy of product reconstitution.Research results: 1?According to the Box-Behnken response surface method analysis,thesignificance of the four protective agents was sucrose content >arginine content > albumin content > sodium chloride content.Theoptimum formulation ratio of optimized design is: sucrose content17.96 g·L-1,arginine content 4.85 g·L-1,sodium chloride content 3.95g·L-1,albumin content 3.79 g·L-1.At this time,the reconstitution timeof the porcine fibrin preparation and the mechanical properties of thefibrin glue are preferred.2?The eutectic point and eutectic point of the porcine fibrinogen formulasolution were determined by electric resistance method to be-16 ?and-12 ?,respectively.The appropriate temperature for thepre-freezing of the Fg preparation was-45 ?,and the pre-freezingtime was compared to determine the pre-freezing time of 3 h.Theinnovation of the process lies in the use of the apparent analyticalinstrument to explore the microstructure,the physical and chemicalproperties of the solids in the lyophilization process,which providesimportant assistance for selecting suitable pre-freezing methods andimproving the product quality.Meanwhile,experimental researchproves that annealing and aeration can be added during thefreeze-drying process.The advantage is that annealing can improve thestructural heterogeneity of the drug,and aeration can reduce theresidual water content and shorten the freeze-drying time.3?The stability and accelerated test of reconstituted porcine fibrinogenlyophilized preparation showed that there was no clot or fibrinprecipitation in the solution after reconstitution in a 37 ? water bathfor 1 h,and 3 batches of small test products were All indicators underthe accelerated test study are in compliance with the regulations. |
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