| Objective:Fresh ginseng is rich in active ingredients such as ginsenosides,proteins,and ginseng polysaccharides.Fresh deer antler contains active substances such as protein peptides,various growth factors,and deer antler polyamines.The combination of the two traditional Chinese medicines has therapeutic and health benefits such as anti-aging,hypoglycemic,anti-fatigue,and improving reproductive function.Currently,the application form of the combination of the two is mostly the preparation made from traditional decoction pieces after processing and extraction.High temperature decoction or reagent extraction cannot effectively reflect the advantages of fresh products.In order to fully utilize the unique therapeutic characteristics of fresh ginseng and antler,this study aims to explore the maximum retention of active substances in fresh medicine by different drying methods,in order to optimize the drying method that does not lose or damage the natural active substances of fresh medicine as much as possible,and to elucidate the potential advantages of freeze-dried shaping preparation technology in preparing fresh medicine oral formulations from the perspectives of chemical composition and biological activity.Finally,a one-step shaping method with high active ingredient content will be prepared,the fresh ginseng and antler freeze-dried oral disintegrating tablets has excellent taste and rapid oral disintegration.Methods:(1)Establish corresponding detection methods for the pharmacological components of ginseng and deer antler.Based on literature research,it was determined that the primary active substance of deer antler is protein peptides.The Bradford method,which is convenient,fast,and highly sensitive,was used to detect the water-soluble protein content of fresh ginseng antler and two different dried products;Ginsenoside is the main active substance contained in ginseng.The content of five ginsenosides in fresh ginseng and two different dried products was determined by HPLC,and the drying method was optimized from the perspective of chemical component retention based on experimental results;(2)Optimize the process parameters and types of excipients for homogenization,set up a freeze-drying process,and preliminarily prepare freeze-drying orally disintegrated tablets.Based on the overall principle of compatibility with fresh ginseng antler,using biological activity indicators that promote TM3 cell proliferation rate,screen the level of process parameters such as homogenization time and type of homogenization diluent during the homogenization preparation process;Comprehensive evaluation indicators such as appearance,hardness,and disintegration time of freeze-dried tablets are used to optimize the types and compounding methods of freeze-dried protective agents through comprehensive scoring;(3)Optimization of the formulation and freeze-drying process for freeze-dried orally disintegrating tablets.Based on the preliminary preparation results of orally disintegrating tablets,the freezedried orally disintegrating tablets were comprehensively evaluated using the taste scoring method.The final formulation of fresh ginseng antler freeze-dried orally disintegrating tablets was optimized using the orthogonal experimental method to determine the final composition of the formula;Using the drying efficiency method combined with the pressure rise method to optimize the temperature and time of the pre freezing stage and two drying stages,and complete the optimization of the freeze-drying preparation process;(4)Basic,chemical,biological,and stability evaluation of freeze-dried oral disintegrating tablets.To verify the stability of the final prescription and preparation process,three batches of freeze-dried orally disintegrated tablets were prepared according to the established prescription and preparation process,and basic evaluation studies were conducted on the appearance,hardness,disintegration time limit,moisture content of the freeze-dried orally disintegrated tablets;The Bradford method was used to determine the content of water-soluble proteins in three batches of orally disintegrated tablets,and the HPLC method was used to conduct chemical evaluation studies on the content of five saponins;Conduct hightemperature,high humidity,and strong light tests on orally disintegrated tablets using stability influencing factor tests,and evaluate the appearance,disintegration time limit,and chemical composition content of freeze-dried orally disintegrated tablet samples at days 0,5,and 10 under three factors;Establish a TM3 oxidative damage cell model,and determine the effects of fresh mixed homogenate supernatant,processed high-temperature decoction extract,and freeze-dried orally disintegrated tablets on the cell viability of the model using CCK-8 method.The SOD enzyme and CAT enzyme activities,cGMP,and NO content of each cell group were also measured,and the in vitro pharmacological activity evaluation of the preparation was preliminarily completed.Results:(1)The results of the determination of pharmacological components showed that fresh ginseng and fresh deer antler dried by vacuum freeze-drying method were more able to retain water-soluble proteins and ginsenoside active ingredients compared with heated drying;(2)The research results of the preparation process showed that when the homogenization time was 6 minutes and 2 times the total dosage of isotonic phosphate buffer was added,the relevant pharmacological components were fully dissolved,which had the best proliferative effect on TM3 cells;When using 10%glycine as the framework agent for freeze-dried orally disintegrating tablets and 1%pullulan as the adhesive,the comprehensive score of freezedried orally disintegrating tablets is the highest and the quality is the best;The fixed skeleton agent and adhesive types,and the results of single factor experiments to simplify the ratio of auxiliary materials show that both types of auxiliary materials can be further simplified,but the glycine content should not be less than 5%,and the pullulan content should not be less than 0.5%;(3)The results of prescription optimization showed that the essence was lemon essence with the lowest proportion of 1%,which was screened by single factor.Combining the lowest addition ratio of glycine and pullulan in the result of excipient simplification,the levels of each factor in the orthogonal experiment were set.According to the range analysis results of the orthogonal experiment,the best prescription combination was A1B2C3,that is,5%glycine,0.6%pullulan and 1.4%lemon essence were added as excipients;The optimization results of the preparation process showed that the pre freezing stage and two drying stages were optimized using the method of drying efficiency of orally disintegrated tablets and combined with the appearance of orally disintegrated tablets.The pre freezing temperature was selected to be-50℃ for 4 hours,the sublimation drying temperature was-10℃ for 5 hours,the analytical drying temperature was 15℃ for 4 hours,and the heating time of both drying temperatures was 1 hour.The final optimized total time for freeze-drying and shaping preparation was 15 hours;(4)In the basic evaluation process,the appearance of orally disintegrating tablets tablets is good,and the inspection items such as hardness,disintegration,and moisture content are all qualified;In the chemical evaluation process,the content of ingredients in the same batch is uniform,and the preparation process of the formulation is stable and reliable;The stability evaluation influencing factors experiment indicates that suitable packaging materials should be selected for this type of fresh drug freeze-drying preparation to maintain the appearance of the preparation and reduce the loss of active ingredients;In the biological activity evaluation process,fresh homogenized supernatant,freeze-dried orally disintegrated tablets,and processed product decoction extracts all had a certain protective effect on the TM3 oxidative damage model.The effect size was as follows:fresh group>tablet group>processed group.Each group enhanced the activity of SOD and CAT enzymes to varying degrees,increased the content of cGMP and NO,and the synchronous increase of cGMP and NO content also indicates that the antioxidant effect of the combination of ginseng and deer antler may be related to the upregulation of the expression of NO-cGMP pathway.Based on the above biological activity evaluation research results,the in vitro therapeutic activity of this preparation has been preliminarily confirmed.Conclutions:This study is based on the freeze-dried shaping preparation platform,combined with low-temperature homogenization,precision filling and other technologies,to prepare a freezedried oral disintegrating tablet of fresh ginseng and fresh deer antler on the same carrier.The medicinal substance of this formulation is stable at room temperature,has a good taste and taste,rapidly disintegrates in the oral cavity,and has a certain degree of biological activity,which can preliminarily meet the demand for compatibility of fresh ginseng antler into medicine,At the same time,it also provides a new direction for the development and utilization of fresh Chinese medicine formulations. |
PDF Full Text Request