Application of panax ginseng extract in preparation of film agent, panax ginseng oral film agent as well as preparation method and application of the same

Abstract

To provide the application of a Panax ginseng extract in the preparation of a film agent, a Panax ginseng oral film agent and a preparation method and application of the same, in the technical field of traditional Chinese medicine.SOLUTION: According to the application of a Panax ginseng extract in preparation of a film agent, a preparation method of the Panax ginseng extract comprises a step of taking an ethanol water solution as an extraction solvent, and performing reflux extraction on Panax ginseng to obtain the Panax ginseng extract, where: a mass ratio of the Panax ginseng to the ethanol water solution is (0.5-1): (6-8); a volume fraction of ethanol in the ethanol aqueous solution is 65-75%; a single extraction time of the reflux extraction is 0.5-1.5 h; and the number of times of the reflux extraction is 1-3.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention belongs to the technical field of traditional Chinese medicine, specifically to the use of ginseng extract in the preparation of a film, ginseng oral film and its preparation method and use. [Background technology]

[0002] Panax ginseng (Panax ginseng CA Mey.) is a perennial plant of the Araliaceae family that is known to have various beneficial effects, including improving memory loss and spatial cognitive ability, as well as anticancer and antidiabetic activity. Its functional benefits also include improving liver function, regulating blood pressure and blood sugar, and providing antioxidant activity. Panax ginseng contains various components such as ginsenosides, acidic polysaccharides, flavones, organic acids, polyacetylenes, alkaloids, and phenolic compounds. Here, ginsenosides such as Rg1, Rb1, Re, Rg3, and Rh2 are the main pharmacologically active components of Panax ginseng, of which Rg1 has more biological activity and has attracted widespread attention.

[0003] Water decoction method and ethanol reflux extraction method are traditional extraction methods. These two extraction methods have the advantages of easy operation and low cost. In the actual production process, they are still commonly used extraction methods in many production enterprises and laboratories. However, in the current extraction process, the extraction rate of ginsenoside Rg1 is poor. Summary of the Invention [Problem to be solved by the invention]

[0004] The present invention aims to provide a use of a ginseng extract in the preparation of a film, a ginseng oral film and a preparation method and use thereof, and in the use provided by the present invention, the obtained ginseng extract Rg1 content is high. [Means for solving the problem]

[0005] To achieve the above objectives, the present invention provides the following technical solutions:

[0006] The present invention provides a use of a Panax ginseng extract in the production of a film preparation, the production method of the Panax ginseng extract comprising: The method includes a step of reflux-extracting ginseng using an aqueous ethanol solution as an extraction solvent to obtain the ginseng extract, The mass ratio of the ginseng to the aqueous ethanol solution is 0.5-1:6-8; The volume fraction of ethanol in the ethanol aqueous solution is 65 to 75%, The extraction time for the reflux extraction is 0.5 to 1.5 hours, and the extraction is performed 1 to 3 times.

[0007] The present invention further provides a ginseng oral film, comprising a backing layer and a drug-containing layer, which are laminated in sequence, the drug-containing layer comprises, by mass, 10-12 parts of hydroxypropyl methylcellulose, 0.1-1 part of carbomer, 9-11 parts of ginseng extract, and 0.1-0.3 parts of fatty acid, the ginseng extract being the ginseng extract described in the above technical solution; The backing layer contains 0.6 to 0.8 parts by mass of ethyl cellulose based on the parts by mass of the hydroxypropyl methyl cellulose.

[0008] Preferably, the fatty acids include one or more of caproic acid, caprylic acid, capric acid, lauric acid, stearic acid, oleic acid and linoleic acid.

[0009] The present invention further provides a method for preparing the ginseng oral film formulation according to the above technical solution, A first injection of ethyl cellulose solution into an anti-adhesive container and a first molding are sequentially performed to obtain a backing layer; Swelling carbomer in water and first mixing with hydroxypropyl methylcellulose to obtain a first film solution; performing a second mixing of a fatty acid solution with the first film liquid to obtain a second film liquid; a third mixing step of mixing the ginseng extract with the second film liquid to obtain a drug-containing layer film liquid; and second-injecting the drug-containing layer film liquid into the backing layer and second-molding the drug-containing layer film liquid into the backing layer to obtain the ginseng oral film.

[0010] Preferably, the temperature of the first mixture is 70 to 90°C.

[0011] Preferably, the density of the ethyl cellulose solution is 0.7 to 0.9 g / cm 3 and The density of the drug-containing layer film liquid is 1.3 to 1.4 g / cm 3 It is.

[0012] Preferably, the temperature of the first molding is 50 to 70° C. and the time is 20 to 40 minutes.

[0013] Preferably, the temperature of the second molding is 60 to 80° C. and the time is 1 to 3 hours.

[0014] The present invention further provides the use of the ginseng oral film formulation described in the above technical solution in the manufacture of an anti-cancer preparation or an anti-diabetic preparation.

[0015] The present invention further provides the use of the ginseng oral film according to the above technical solution in the manufacture of a blood pressure regulating preparation, a blood glucose regulating preparation or an antioxidant preparation.

[0016] The present invention provides a use of ginseng extract in the preparation of a film preparation, the preparation method of the ginseng extract includes the steps of using an aqueous ethanol solution as an extraction solvent, refluxing ginseng to obtain the ginseng extract, the mass ratio of the ginseng to the aqueous ethanol solution is 0.5-1:6-8, the volume fraction of ethanol in the aqueous ethanol solution is 65-75%, the single extraction time of the reflux extraction is 0.5-1.5h, and the number of times is 1-3. The present invention uses an aqueous ethanol solution as an extraction solvent, effectively extracts ginsenoside Rg1 by reflux extraction, and limits the appropriate parameter range so that the content of ginsenoside Rg1 in the obtained ginseng extract is high, thereby reducing the cost of raw materials and being suitable for mass production. Effect of the Invention

[0017] The ginseng oral film provided by the present invention can directly act on the oral buccal mucosa, which has a weaker barrier function than the skin, and can avoid the inhibitory effect of the skin stratum corneum on the penetration of ginsenoside and the first-pass effect, and can also play a role in improving the bioavailability of Rg1. At the same time, the ginseng oral film is smooth and flat, free of drug crystals, has good mechanical properties, good dissolution rate, is easy to take, and has good patient compliance. [Brief description of the drawings]

[0018] In order to more clearly describe the embodiments of the present invention or the technical solutions of the prior art, the drawings necessary to be used in the embodiments are briefly described below. Of course, the drawings described below are only a part of the embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without creative efforts.

[0019] [Figure 1] 1 shows the appearance of the ginseng oral film preparation described in Example 2. [Diagram 2] 1 shows the dissolution rate of the ginseng oral film preparation described in Example 2 in artificial saliva. [Diagram 3] This shows the plasma concentration-time curves of Rg1 following intragastric and oral buccal administration. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The present invention provides a use of a Panax ginseng extract in the production of a film preparation, the production method of the Panax ginseng extract comprising: The method includes a step of reflux-extracting ginseng using an aqueous ethanol solution as an extraction solvent to obtain the ginseng extract, The mass ratio of the ginseng to the aqueous ethanol solution is 0.5-1:6-8; The volume fraction of ethanol in the ethanol aqueous solution is 65 to 75%, The extraction time for the reflux extraction is 0.5 to 1.5 hours, and the extraction is performed 1 to 3 times.

[0021] In the present invention, unless otherwise specified, all of the above-mentioned raw material ingredients are commercially available products well known to those skilled in the art.

[0022] In the present invention, ginseng is reflux-extracted using an aqueous ethanol solution as an extraction solvent to obtain the ginseng extract.

[0023] In the present invention, the mass ratio of the ginseng to the aqueous ethanol solution is 0.5 to 1:6 to 8, preferably 1:6 to 8, and more preferably 1:7 to 8. The volume fraction of ethanol in the ethanol aqueous solution is 65 to 75%, preferably 67 to 73%, and more preferably 69 to 71%. The extraction time for a single reflux extraction is 0.5 to 1.5 hours, preferably 0.7 to 1.3 hours, and more preferably 0.9 to 1.1 hours, and the number of times is 1 to 3 times, and more preferably 2 to 3 times.

[0024] The use of the ginseng extract provided by the present invention in the production of film preparations uses an aqueous ethanol solution as the extraction solvent, effectively extracts ginsenoside Rg1 by reflux extraction, and limits the range of suitable parameters to ensure that the content of ginsenoside Rg1 in the obtained ginseng extract is high, thereby reducing raw material costs and making it suitable for mass production.

[0025] The present invention further provides a ginseng oral film, comprising a backing layer and a drug-containing layer, which are laminated in sequence, the drug-containing layer comprises, by mass, 10-12 parts of hydroxypropyl methylcellulose, 0.1-1 part of carbomer, 9-11 parts of ginseng extract, and 0.1-0.3 parts of fatty acid, the ginseng extract being the ginseng extract described in the above technical solution; The backing layer contains 0.6 to 0.8 parts by mass of ethyl cellulose based on the parts by mass of the hydroxypropyl methyl cellulose.

[0026] The drug-containing layer contains, in terms of parts by mass, 10 to 12 parts, preferably 11 to 12 parts, of hydroxypropyl methylcellulose.

[0027] The drug-containing layer contains 0.1 to 1 part, preferably 0.3 to 0.8 parts, and more preferably 0.5 to 0.7 parts of carbomer based on the parts by mass of the hydroxypropyl methylcellulose.

[0028] The drug-containing layer contains 9 to 11 parts, preferably 10 to 11 parts, of ginseng extract based on the parts by mass of the hydroxypropyl methylcellulose.

[0029] The drug-containing layer contains 0.1 to 0.3 parts of fatty acid, and preferably 0.2 to 0.3 parts, based on the parts by mass of the hydroxypropyl methylcellulose.

[0030] In the present invention, the fatty acid preferably comprises one or more of caproic acid, caprylic acid, capric acid, lauric acid, stearic acid, oleic acid and linoleic acid, more preferably comprises one or more of caprylic acid, capric acid, oleic acid and linoleic acid, and most preferably comprises capric acid. When the fatty acid comprises two or more of the above specific selections, the present invention does not particularly limit the blending ratio of the fatty acid.

[0031] In the present invention, the thickness of the drug-containing layer is preferably 1.45 to 1.5 mm, more preferably 1.45 to 1.48 mm, and most preferably 1.45 mm.

[0032] The backing layer contains 0.6 to 0.8 parts, preferably 0.7 to 0.8 parts, of ethyl cellulose based on the parts by mass of the hydroxypropyl methyl cellulose.

[0033] In the present invention, the thickness of the backing layer is preferably 0.6 to 0.8 mm, more preferably 0.6 to 0.7 mm, and most preferably 0.6 mm.

[0034] The ginseng oral film provided by the present invention can directly act on the oral buccal mucosa, which has a weaker barrier function than the skin, and can avoid the inhibitory effect of the skin stratum corneum on the penetration of ginsenoside and the first-pass effect, and can also play a role in improving the bioavailability of Rg1. At the same time, the ginseng oral film is smooth and flat, free of drug crystals, has good mechanical properties, good dissolution rate, is easy to take, and has good patient compliance.

[0035] The present invention further provides a method for preparing the ginseng oral film formulation according to the above technical solution, A first injection of ethyl cellulose solution into an anti-adhesive container and a first molding are sequentially performed to obtain a backing layer; Swelling carbomer in water and first mixing with hydroxypropyl methylcellulose to obtain a first film solution; performing a second mixing of a fatty acid solution with the first film liquid to obtain a second film liquid; a third mixing step of mixing the ginseng extract with the second film liquid to obtain a drug-containing layer film liquid; and second injecting the drug-containing layer film liquid into the backing layer and second molding the drug-containing layer film liquid into the backing layer to obtain the ginseng oral film.

[0036] In the present invention, the ethyl cellulose solution is first injected into the adhesion prevention container and then the first molding is carried out to obtain the backing layer.

[0037] In the present invention, the concentration of the ethyl cellulose solution is preferably 0.01 to 0.04 g / mL, more preferably 0.015 to 0.035 g / mL, and most preferably 0.02 to 0.03 g / mL. The density of the ethyl cellulose solution is preferably 0.7 to 0.9 g / cm. 3 , more preferably 0.75 to 0.85 g / cm 3 , and most preferably 0.80 to 0.83 g / cm 3 and the solvent of the ethyl cellulose solution is preferably ethanol, more preferably absolute ethanol.

[0038] In the present invention, the temperature of the first molding is preferably 50 to 70°C, more preferably 55 to 65°C, and most preferably 60 to 65°C, the time is preferably 20 to 40 minutes, more preferably 25 to 35 minutes, and most preferably 30 to 35 minutes, and the first molding is preferably performed by a vacuum drying method.

[0039] In the present invention, after obtaining the backing layer, carbomer is swelled in water and mixed with hydroxypropylmethylcellulose to obtain a first film liquid.

[0040] In the present invention, the mass ratio of the carbomer to water is preferably 1-4:80-130, more preferably 1-3:90-120, and most preferably 1-2:100-103.

[0041] In the present invention, the temperature of the first mixing is preferably 50 to 70°C, more preferably 55 to 65°C, and most preferably 60 to 65°C.

[0042] In the present invention, after obtaining the first film liquid, a second mixing of the fatty acid solution and the first film liquid is carried out to obtain a second film liquid.

[0043] In the present invention, the concentration of the fatty acid solution is preferably 0.4 to 0.5 g / mL, more preferably 0.42 to 0.48 g / mL, and most preferably 0.43 to 0.47 g / mL, and the solvent for the fatty acid solution is preferably propylene glycol.

[0044] The present invention does not impose any particular limitation on the second mixing step, and any method well known to those skilled in the art may be employed.

[0045] In the present invention, after obtaining the second film liquid, a third mixing of the ginseng extract and the second film liquid is carried out to obtain a drug-containing layer film liquid.

[0046] In the present invention, the density of the drug-containing layer film liquid is preferably 1.3 to 1.4 g / cm 3 , more preferably 1.31 to 1.35 g / cm 3 , and most preferably 1.31 to 1.32 g / cm 3 and In the present invention, the third mixing is preferably performed at room temperature. The present invention does not impose any particular limitations on the process of the third mixing, and any method well known to those skilled in the art may be used.

[0047] In the present invention, after obtaining the drug-containing layer film liquid, the drug-containing layer film liquid is sequentially injected into the backing layer and then molded to obtain the ginseng oral film preparation.

[0048] In the present invention, the temperature of the second shaping is preferably 60 to 80°C, more preferably 65 to 75°C, and most preferably 70 to 75°C, the time is preferably 1 to 3 hours, more preferably 1.5 to 2.5 hours, and most preferably 2 to 2.5 hours, and the second shaping is preferably performed by a vacuum drying method.

[0049] The manufacturing method provided by the present invention is convenient, rapid, and easy to control the quality.

[0050] The present invention further provides the use of the ginseng oral film formulation described in the above technical solution in the manufacture of an anti-cancer preparation or an anti-diabetic preparation.

[0051] The present invention further provides the use of the ginseng oral film according to the above technical solution in the manufacture of a blood pressure regulating preparation, a blood glucose regulating preparation or an antioxidant preparation.

[0052] In order to further illustrate the present invention, the use of the ginseng extract in the preparation of a film preparation provided by the present invention, the ginseng oral film preparation and its preparation method and use are described in detail below with reference to the drawings and examples, which should not be construed as limiting the protection scope of the present invention.

[0053] Example 1 The ginseng extract was extracted under reflux using an aqueous ethanol solution with a volume fraction of 70% as the extraction solvent at a solid-liquid ratio of 1:8, extraction time of 1 hour, and extraction times of 2 times to obtain the ginseng extract.

[0054] Example 2 Ethyl cellulose was dissolved in absolute ethanol to obtain an ethyl cellulose solution with a concentration of 0.025 g / mL and 0.81 g / cm 3 The mixture was poured into a petri dish at a density of 1000×1000 and dried in vacuum at 60° C. for 30 minutes to obtain a backing layer. 0.786 g of carbomer was swollen in water for 12 h, and the film solution was heated to 90°C in a water bath. Then, 10.742 g of hydroxypropyl methylcellulose was added and dissolved for 1 h to obtain a first film solution. Capric acid was dissolved in propylene glycol to obtain a capric acid solution with a concentration of 0.47 g / mL, which was added to the first film solution to obtain a second film solution. After the second film solution was cooled to room temperature, 10.155 g of ginseng extract was added and stirred until dissolved, and the density of the resulting solution was 1.31 g / cm. 3 Thus, a drug-containing layer film solution was obtained. The drug-containing layer film liquid was poured into the backing layer and dried in vacuum at 70°C for 2 hours to obtain a ginseng oral film preparation.

[0055] Test Example 1 Establishment of a method for extracting ginseng 1.1 Reagents and Equipment 1.1.1 Experimental materials and reagents The materials and reagents used in this experiment are shown in Table 1-1. [Table 1] 1.1.2 Experimental equipment The experimental equipment used in this experiment is shown in Table 1-2. [Table 2]

[0056] 1.2 HPLC content measurement method of Rg1 The content of Rg1 in the extracts was measured using high performance liquid chromatography. 1.2.1 Chromatographic conditions The chromatographic conditions for measuring Rg1 by HPLC are shown in Tables 1-3. [Table 3] 1.2.2 Preparation of solutions Standard substance solution: 5.0 mg of the Rg1 standard substance was weighed out and the volume was adjusted to 10 mL with methanol to prepare a solution of Rg10.5 mg / mL, which was then diluted to the corresponding concentration when used. Test substance solution: 100.0 mg of the dried extract of Panax ginseng was weighed and placed in a 10 mL measuring flask, 5 mL of methanol was added, and the mixture was ultrasonicated for 30 minutes, allowed to cool, and methanol was added up to the mark to obtain 10 mL of test substance solution. 1.2.3 Results of specificity experiments Specificity Results The retention time of Rg1 is 8.10 min, and the chromatographic peak shape of the component is good and well separated from the solvent peak and other components, so this chromatographic condition can be used to measure the content of Rg1. 1.2.4 Results of methodological considerations The methodological considerations for the determination of the content of Rg1 are shown in Tables 1-4 and 1-5. The results show that the established HPLC method is reliable and can be used to determine the content of Rg1 in ginseng extracts. [Table 4] [Table 5]

[0057] 1.3 Analysis of orthogonal test results 1.3.1 Results analysis of the content of Rg1 in the extracts L9(3 3 ) Extraction was performed according to the orthogonal design test table, the three factors being the solid-liquid ratio, extraction time, and number of extractions, respectively, and the obtained extract was freeze-dried to obtain an extract. The extract was weighed, and the content of Rg1 in the extract was measured by HPLC, the results are shown in Tables 1-6 and 1-7, and the analysis of variance tables are shown in Tables 1-8 and 1-9. The analysis of variance results show that none of the factors have a significant effect on the content of Rg1 in the extract. The extraction process with the highest content of extract Rg1 in the water decoction method is A1B1C1, and the influencing factors are R C >R B >R A The extraction process with the highest content of Rg1 in the extract in the ethanol reflux extraction method is A2B2C2, and the influencing factors are R A >R B >RC It is. [Table 6] [Table 7] R 2 =0.933 [Table 8] [Table 9] R 2 =0.744

[0058] 1.4 Summary The water decoction method and the ethanol reflux extraction method are traditional extraction methods, and these two kinds of extraction methods have the advantages of easy operation and low cost. In the actual production process, they are still commonly used extraction methods in many production enterprises and laboratories. In order to increase the extraction rate of Rg1, it is necessary to select the optimal extraction process suitable for actual production. The results of the orthogonal design test of the water decoction method show that with the increase of extraction time and number of extractions, the extraction rate of Rg1 decreases inversely. On the one hand, Rg1 may be decomposed by long-term heating, and on the other hand, compounds such as ginseng polysaccharides, which have higher solubility in water, have a higher extraction rate in water. Summarizing the experimental results, the optimal extraction process for producing ginseng extract is to use an ethanol aqueous solution as an extraction solvent, reflux extract ginseng to obtain the ginseng extract, the mass ratio of the ginseng to the ethanol aqueous solution is 0.5-1:6-8, the volume fraction of ethanol in the ethanol aqueous solution is 65-75%, the single extraction time of the reflux extraction is 0.5-1.5 hours, and the number of times is 1-3.

[0059] Test Example 2 Study on the permeability of fatty acids in ginseng extract This test example mainly examined the penetration-promoting effect of fatty acids on Rg1, and used Rg1 in the ginseng extract as the indicator component. Five types of saturated fatty acids, namely caproic acid (C6), caprylic acid (C8), capric acid (C 10 ), lauric acid (C 12 ), stearic acid (C 18 ), two unsaturated fatty acids, namely oleic acid (C 18:1 ), linoleic acid (C 18:2 The penetration enhancing effect of fatty acids was discussed, and the indexes considered included steady-state flux, apparent permeability coefficient, permeation enhancement factor, lag time and 4h cumulative permeation amount. The permeation enhancing effect of different fatty acids was evaluated using principal component analysis, providing a reference basis for the rational use of permeation enhancers in ginseng oral film.

[0060] 2.1 Reagents and Equipment 2.1.1 Experimental materials and reagents The materials and reagents used in this experiment are shown in Table 2-1. [Table 10] 2.1.2 Experimental equipment The experimental equipment used in this experiment is shown in Table 2-2. [Table 11]

[0061] 2.2 Method 2.2.1 Preparation of solutions 110.0 mg of Rg1 standard was weighed out and dissolved in 10 mL of PBS solution at pH=6.8, shaken for 1 h, and sonicated for 1 h to prepare 10 mL of ginseng extract supply solution containing the same concentration (11 mg / mL) of Rg1. The supply solution was stored at 4°C for use, and the supply solution was vortexed for 5 min before use. 2.2.2 Preparation of fatty acid permeation enhancer solution Seven types of fatty acids, namely caproic acid, caprylic acid, capric acid, lauric acid, stearic acid, oleic acid and linoleic acid, were dissolved in 95% ethanol to obtain penetration enhancer solutions with respective concentrations (1-10 mg / mL). 2.2.3 In vitro permeation experiments The treated porcine esophageal epithelium was then placed in a Franz diffusion cell (diffusion area 1.76 cm 2 The receiving chamber was sandwiched in the center of a solution volume of 4.5 mL. The outer layer of the porcine esophageal epithelium was facing the supply chamber, and the inner layer of the porcine esophageal epithelium was facing the receiving chamber, and 1.0 mL of PBS solution with a pH of 6.8 was added to the supply chamber. After 5 min, the receiving chamber was checked for leakage of the solution in the supply chamber due to damage to the porcine esophageal epithelium or improper installation of the diffusion cell. After confirming that there was no abnormality, the solution in the supply chamber was discarded and wiped off. 40.0 μL of the permeation enhancer solution was added to the supply chamber, and the supply chamber was left open for 1 h to ensure the ethanol solvent evaporates and dries, and the interference of the solvent was removed. 4.5 mL of pH=7.4 PBS solution was added to the receptor chamber, 0.2 mL of the supply solution was added to the supply chamber, the Franz diffusion cell was placed in the transdermal diffusion apparatus, a magnetic stirrer was added, the rotation speed was set to 600 rpm, the temperature was set to 37°C±0.5°C, 0.3 mL of the receptor solution was aspirated from the receptor chamber at 0, 0.25, 0.5, 0.75, 1, 1.5, 2.5, 3, and 4 h, respectively, and the receptor chamber was refilled with 0.3 mL of fresh isothermal pH=7.4 PBS buffer. 2.2.4 Data Analysis The measured porcine esophageal epithelium permeability data are the apparent permeability coefficient P, the lag time TL, and the steady-state flow rate J, and the calculation methods are shown in Equation 1, Equation 2, Equation 3, and Equation 4.

number

number

number

number

[0062] 2.3 Results 2.3.1 Penetration-enhancing effect of seven fatty acids on Rg1 During the permeation experiment, the concentrations of the receptor fluids were all allowed to reach the sink condition, and the drug-containing concentrations of the receptor fluids at different time points were measured. The data was subjected to calculation processing to obtain the steady-state flux of Rg1 permeating through the porcine esophageal epithelium, the apparent permeability coefficient, the permeation enhancement factor, the 4-h cumulative permeation amount, the lag time and other parameters, which are shown in Tables 2-3. The results show that the cumulative permeation amount of Rg1 has a good linear relationship with time. Compared with the blank group, the drug concentration of 1 mg / mL and 5 mg / mL stearic acid (C 18 ) showed no significant difference in ER (p>0.05), proving that 1mg / mL and 5mg / mL stearic acid had no permeation enhancing effect on Rg1. The remaining fatty acids could all increase the permeability coefficient of Rg1 to different degrees, and among the saturated fatty acids, capric acid (C 10 ) had the highest permeation enhancement factor, with an ER of 19.24 ± 1.89. Among unsaturated fatty acids, the highest permeation enhancement factor was 5 mg / mL linoleic acid (C18:2 ) and ER: 9.40±1.00. [Table 12] 2.3.2 Analysis of the effect of penetration enhancers on Rg1 using principal component analysis (1) Experiments on standardization of permeation data and applicability of factor analysis The data in Table 2-3 is substituted into IBM SPSS Statistics 26.0 software, standardization processing is performed, the unit effect of different indicators is eliminated, and standardized data is obtained, and then the applicability experiment of factor analysis is performed, and the results are shown in Table 2-4. When using principal component analysis to conduct information enrichment research, it is necessary to first analyze whether the data is suitable for principal component analysis, and the KMO value is equal to 0.797, which indicates that there is a certain correlation between the five indicators, and in the Bartelett sphericity test, the Sig value is 0.000 (p<0.05), which rejects the correlation coefficient as a unit matrix, that is, each indicator is related, and the two result tests indicate that the data is suitable for principal component analysis. [Table 13] (2) Calculation of principal component feature vectors The eigenvalue and variance contribution rate of the correlation coefficient were calculated by SPSS software, and the results are shown in Table 2-5. The contribution rates of component 1 and component 2 were 81.66% and 18.13%, respectively, and the cumulative contribution rate was 99.78%>85.00%, which indicates that component 1 and component 2 can represent 99.78% of the five index information. This indicates that component 1 and component 2 can represent the comprehensive judgment of the penetration enhancing effect of the penetration enhancer on Rg1. [Table 14] Component 1 = 0.244 × steady-state flux + 0.244 × apparent permeability coefficient + 0.2444 × permeability enhancement factor + 0.243 × 4-h cumulative permeation + 0.085 × lag time; Component 2 = -0.085 x steady state flux -0.085 x apparent permeability coefficient -0.085 x permeability enhancement factor -0.107 x 4h cumulative permeation + 1.035 x lag time. (3) Data calculation of principal component scores and total scores The standardized data of each processed variable was substituted into a linear equation, and the total score was cumulatively calculated by multiplying the variance explanation rate and the component score. The formula for the total score was total score = (81.66% × component 1 + 18.12% × component 2) / 99.783%, and the total score was sorted. The results are shown in Table 2-6. From the ranking in the table, it was found that the order was 1% capric acid > 5% capric acid > 5% linoleic acid > 1% linoleic acid > 1% oleic acid > 5% oleic acid > 10% capric acid > 10% oleic acid > 10% linoleic acid > 10% caproic acid > 1% caprylic acid > 1% caproic acid > 5% caproic acid > 1% lauric acid > 5% lauric acid > 10% stearic acid > 10% lauric acid > 1% stearic acid > 5% stearic acid > blank group > 5% caprylic acid > 10% caprylic acid, and from the overall score, it was found that 1% capric acid had the highest penetration enhancing effect. [Table 15]

[0063] 2.4 Summary Fatty acids can increase the ability of Rg1 to penetrate through the porcine esophageal epithelium, with a concentration of 0.227 mg / cm 2 The penetration enhancing effect was greatest when capric acid acted on the porcine esophageal epithelium.

[0064] Test Example 3: Preparation study of ginseng oral film 3.1 Reagents and Equipment 3.1.1 Experimental materials and reagents The materials and reagents used in this experiment are shown in Table 3-1. [Table 16] 3.1.2 Experimental equipment The experimental equipment used in this experiment is shown in Table 3-2. [Table 17]

[0065] 3.2 Method The ginseng oral film is made into a two-layer film, one of which is a water-insoluble backing layer, which can control the unidirectional release of the drug, as well as protect the drug-containing layer from saliva erosion. The other layer is the drug-containing layer, which is made of a hydrophilic polymer material with a certain in vitro adhesive force, which can ensure that the film adheres to the surface of the buccal mucosa and maintain the penetration of the drug. 3.2.1 Selection of film formation method Common methods for the production of film preparations include coating, solvent injection, composite film formation, hot melt extrusion, thermoplastic film formation, semi-solid solvent injection, solid dispersion extrusion and spheronization. In production, the coating and solvent injection methods are generally used, and these two film formation methods are easy to operate, dry quickly and have a high production rate. The coating method is difficult to control the thickness, the drug loading is too low, and it is not suitable for the production method of herbal medicine film preparations. The solvent injection method pours the adhesive solution into a container of a certain volume, and the amount and area of ​​the film formation material used can be controlled. It is suitable for the production of herbal medicine film preparations in the laboratory. 3.2.2 Selection of drug-containing layer film-forming materials The film agent is essentially a polymer matrix, composed of one or more polymers, and currently commonly used materials include gelatin, shellac, carboxymethylcellulose, pullulan, hydroxypropylcellulose, hydroxypropylmethylcellulose, sodium alginate, etc. Hydroxypropylmethylcellulose has excellent mechanical properties and drug-carrying capacity after film formation, so this subject selects hydroxypropylmethylcellulose as the main film-forming material, and since the cellulose derivative has low mucoadhesion when used alone, an appropriate amount of carbomer is added as an adhesive to improve the bioadhesion of the drug carrier, and at the same time, the combination of carbomer and hydroxypropylmethylcellulose can serve as a sustained-release framework material for the film agent to maintain drug release. 3.2.3 Selection of Plasticizer for Drug-containing Layer The film preparation made by using only the film-forming material has a hard texture and poor mechanical properties, while the addition of a plasticizer can improve the flexibility of the film preparation and reduce the brittleness of the film preparation, which is favorable for the transportation and storage of the film preparation.When glycerin is used as the plasticizer, adhesion occurs between the film preparation, which is unfavorable for the storage of the film preparation.When triethyl citrate is used as the plasticizer, the surface of the film preparation is smooth, but the brittleness is high and the flexibility is low.When propylene glycol is used as the plasticizer of this formulation, the surface of the film preparation is smooth and the flexibility is appropriate, which is suitable for the plasticizer of this formulation. 3.2.4 Selection of backing layer material As the main drug, ginseng is easily dissolved in water, and ginseng extract can be uniformly dispersed in the film, but ginseng is bitter, which reduces the patient's compliance and is unfavorable to taking the oral film. At the same time, the single-layer drug-containing film is eroded by saliva, swells and disintegrates quickly, and cannot be retained in the oral cavity for a long time. In consideration of the patient's compliance and oral retention time, a backing layer is added to the drug-containing layer to produce a two-layer oral film. The backing layer requires that the selected film-forming material is both insoluble in water at human body temperature and tightly adheres to the drug-containing layer. Finally, ethyl cellulose is selected as the backing layer material. 3.2.5 Consideration method (1) Appearance of the film The appearance requirements of the film are that the appearance is complete and smooth, the thickness is consistent, the color is uniform, and there are no obvious bubbles. For multi-dose films, the dividing impression is uniform and clear, and the impression can be pressed and torn. (2) Film solution Although the Pharmacopoeia does not require the film solution to have proper fluidity, the solvent injection method requires that the film solution has appropriate fluidity. If the film solution is too viscous, it is difficult to eliminate air bubbles and to pour it out of the container, and if the film solution has too high fluidity, it takes a long time to dry and it is difficult to control the quality of the produced film. The adhesive solution system should be homogeneous, with no obvious insoluble lumpy solids visible to the naked eye as a criterion. (3) Dissolution time The oral film (2.2 cm x 3.2 cm) was placed on top of a petri dish, and 200 μL of artificial saliva (KH2PO4 12 mM, NaCl 40 mM, CaCl2 21.5 mM, H2O 1 L, pH value adjusted to 6.8) at 37°C was dropped onto the film until the droplet penetrated the film and dropped onto the petri dish. The time it took for the droplet to penetrate the film and drop to the bottom of the petri dish was measured and recorded as the dissolution time. Each group of samples was measured six times, and the average value was recorded. (4) In vitro adhesion The oral film agent is administered to the oral buccal mucosa in this specification, which requires that the oral film agent adheres to the oral buccal mucosa tissue to release Rg1 and penetrate the buccal mucosa to enter the blood circulation, which requires that the film agent has a certain adhesive force to the oral mucosa. A slide glass is fixed to the base using an Adelbo HLB universal material testing machine. A clip is fixed to the center point of the slide glass using another slide glass, and the prepared slide glass with clip is fixed to the upper clip of the universal material testing machine, and the two slide glasses are horizontal. Next, the prepared film agent isolation layer was fixed to a clipped glass slide at the top with double-sided tape, 200 μL of 5% (w / w) mucin solution was dropped onto the base glass slide, the upper glass slide and the lower glass slide were brought into close contact, a 2 g weight was placed on top of them for 30 s, and then the glass slide was pulled upward at a constant speed of 0.1 mm / s. At the moment when the two parts of the glass slide were pulled apart, the tension reading displayed on the universal testing machine at that time was recorded as the in vitro adhesive strength of the film agent.

[0066] 3.3 Results 3.3.1 Optimization of blank film formulation using Bok-Behnken response surface methodology The experimental design and results are shown in Table 3-3. The experimental data were analyzed using Design Expert 12, and multiple nonlinear regression fitting was performed for the three selected factors, with the dissolution time of the blank film formulation and the in vitro adhesive strength as evaluation indicators (Y1) and (Y2). The software was used to create contour plots and three-dimensional response surface plots for the two indicators and the three factors. [Table 18] Using Design Expert 12 software, statistical processing was performed on the data in the table, and the multiple nonlinear fitting equations for fitting each factor to the dissolution time and the associated coefficients are respectively as follows: Y=122.54+16.68A+86.64B-16.07C-17.27AB-11.78AC-10.95BC-20.41A 2 +36.68B 2 -52.30C 2 , R = 0.9674. The results of the analysis of variance are shown in Table 3-4. As can be seen from Table 3-4, p<0.05 is considered as a significant difference, indicating that the nonlinear model used has significance. At the same time, the value of the lack of fit term is not significant when it is greater than 0.05, indicating that the equation fits the experiment well. Here, B, B 2 , C 2 has a significant effect on Y1. [Table 19] The multiple nonlinear fitting equations for fitting the in vitro adhesion forces to each factor and the associated coefficients are respectively as follows: Y=7.28+0.7471A+2.85B+1.11C-0.6417AB-0.0392AC+0.58785BC+0.0695A 2 -1.08B 2 +0.4656C 2, R = 0.9723. The results of the analysis of variance are shown in Table 3-5. A p<0.05 is considered a significant difference, indicating that the nonlinear model used is significant. At the same time, a value of the lack of fit term greater than 0.05 is not significant, indicating that the equation fits the experiment well. Here, A, B, C, B 2 has a significant effect on Y2. [Table 20] Next, the analysis module of Design Expert 10.0 software was used to create three-dimensional effect surface and contour plots for the total score of the interaction of each factor, and it was found that the amount of carbomer (A), HPMC (B), and propylene glycol (C) had some effect on the dissolution time and in vitro adhesive strength of the blank film formulation. After processing the experimental results with Design-Expert 12 software, the longest dissolution time and the maximum in vitro adhesive strength were used as evaluation indexes, and the formulation with the optimized film formulation, i.e., carbomer: 0.1-1g, HPMC: 1-5g, and propylene glycol: 0.1-0.3g, was obtained. Three validation tests were conducted according to the optimized formulation, and the average dissolution time and average in vitro adhesion force obtained were 225.58 s and 9.63 N, respectively, with relative errors from the predicted values ​​of 0.48% and 0.31%, indicating that the formulation was stable and reliable. 3.3.2 Consideration of drug loading When the ginseng extract accounted for 25%, 45%, and 65% of the solid content, respectively, highlight crystals appeared in the field of view when the ginseng extract accounted for 65% of the solid content, and finally, the drug loading was determined to be 45% of the solid content. 3.3.3 Selection of drying equipment According to the optimized formula, the drying method for producing ginseng oral film was considered. According to the existing conditions of the laboratory, three drying methods were selected: vacuum drying method, hot air drying box drying method, and hot plate drying method. They were selected according to the appearance evaluation index of the pharmacopoeia. The experimental results are shown in Table 3-6. The vacuum drying method uses the principle of vacuum negative pressure to lower the boiling point of water and accelerate the evaporation of water in the film liquid, and after drying, the surface of the film is smooth, flat and uniform, without bubbles; the hot air drying box drying method fully fills the drying box with hot air flowing through it to remove the water in the film liquid, and after drying, the hot air blows on the surface of the film liquid, so wrinkles occur on the surface of the film, and the film is not flat, which does not meet the requirements for the appearance of the film in the pharmacopoeia; the heating device in the equipment is not evenly arranged, and a petri dish is sandwiched between the heating plate and the film liquid, so the heating is not uniform when the film liquid is heated, and the thickness of the film is different. Therefore, the vacuum drying method was selected as the drying method for the film liquid. [Table 21] 3.3.4 Selection of drying temperature and time The results of selecting the drying temperature and time are shown in Table 3-7. The drying method was selected as vacuum drying, and the minimum moisture content of the ginseng oral film was used as the selection criterion for the drying temperature and drying time. When the temperature was 70℃, there was no significant difference between the data for 2h and 3h (p>0.05), and finally, in order to save the drying time and cost, the drying temperature of 70℃ and the drying time of 2h were selected. [Table 22] 3.3.5 Optimal formulation The formulation of the ginseng oral film is shown in Table 3-8. The main ingredient of the ginseng oral film is ginseng extract, capric acid is the penetration enhancer, HPMC is the main film-forming material, carbomer is the adhesive, and propylene glycol is the plasticizer. [Table 23] 3.3.6 Optimal manufacturing process First, the carbomer was allowed to swell for 12 hours, and the film solution was heated to 90°C in a water bath. Hydroxypropylmethylcellulose was then added and dissolved for 1 hour, and capric acid was dissolved in propylene glycol and added to the film solution. After the film solution was cooled to room temperature, ginseng extract was added, stirred until dissolved, and left to stand for use. Ethylcellulose was dissolved in absolute ethanol to give a concentration of 0.81 g / cm. 3 The film was poured into a petri dish at a density of 1000 ml and dried in vacuum at 60℃ for 30 min to prepare a backing layer. The drug-containing layer film solution was then poured into the backing layer and dried in vacuum at 70℃ for 2 h to obtain a ginseng oral film preparation. Rg1 is thermally unstable and decomposes with increasing heating time, so it is necessary to add the ginseng extract after cooling to room temperature during the manufacturing process.

[0067] Test Example 4 Quality Study of Ginseng Oral Film This test example was an initial study on the quality of a ginseng oral film formulation. 4.1 Reagents and Equipment 4.1.1 Experimental materials and reagents The materials and reagents used in this experiment are shown in Table 4-1. [Table 24] 4.1.2 Experimental equipment The experimental equipment used in this experiment is shown in Table 4-2. [Table 25]

[0068] 4.2 Method 4.2.1 Visual Inspection The ginseng oral films were visually observed for obvious air bubbles, deformation and uneven color. 4.2.2 Microscopic inspection A cold field emission scanning electron microscope (SU 8010) was used to impact the surface of the oral film under high vacuum and electron beam irradiation to obtain the microscopic structural morphology of the oral film surface, including whether the drug crystals were present on the surface and whether the structure was intact, and the obtained data were mutually corroborated with other properties of the oral film, such as mechanical properties and dissolution rate. A blank oral film without drug and a ginseng oral film were placed in a vacuum drying box at 40℃ for 12 hours to ensure the evaporation of water in the oral film. The dried oral film was attached to a metal stand by a conductive adhesive, and gold was sprayed on the surface of the film to obtain good conductivity, and the microscopic structural morphology of the two oral film surfaces was examined. 4.2.3 Weight Difference Limit Inspection Twenty tablets of each ginseng oral film preparation were taken, and the total weight was precisely weighed to determine the average weight. Then, the weight of each tablet was precisely weighed, and the weight of each tablet was compared with the average weight to check for weight differences. 4.2.4 In vitro residence time studies In order to simulate the time that the oral film can remain in the oral cavity after being affected by mechanical factors such as saliva flow and washing, the following device was specially manufactured: The upper part is a linear reciprocating mechanical device, a glass slide is fixed to a telescopic lever, and 5% (w / w) mucin solution is sufficiently applied to the glass slide to simulate the surface of the oral mucosa, a blank oral film without drug and a drug-containing layer of a ginseng oral film are attached to the surface, a 2g weight is placed on the surface of the film for 30s, and the lower part is artificial saliva as the test medium, the glass slide moves up and down, and one time is calculated as the time when the upper edge of the film is immersed in the artificial saliva and the lower edge is separated from the artificial saliva, the mechanical device is set to 60 times / min, and the time when the film is completely separated from the glass slide is recorded to simulate the retention time of the oral film in the oral cavity. 4.2.5 Tensile strength test According to the requirements of the People's Republic of China national standard GBT 1040.2-2006, the blank oral film without drug loading and the ginseng oral film were cut into a dumbbell shape with a dumbbell cutter, and the cut films were fixed in a WDT-10 electronic universal testing machine with a gauge distance of 40 mm between two clamps, and the oral films were stretched in the longitudinal direction at a speed of 50 mm / min until they broke, and six samples were measured. The calculation formulas are shown in Equation 5 and Equation 6.

number

number

[0069] 4.3 Results 4.3.1 Appearance of the film The appearance of the ginseng oral film is shown in Figure 1, which shows that the obtained ginseng oral film is complete and smooth, has a consistent thickness, is uniform in color, has no obvious bubbles, has a light yellow surface, and has no special odor. 4.3.2 Microscopic Inspection The results were examined under an electron microscope. The films were placed under a cold-field emission scanning electron microscope and observed under a magnification of 1000 times. The surfaces of the blank oral film and the ginseng oral film, which did not contain any drug, were dense, smooth and flat, without any pores, bubbles, wrinkles or drug crystals, and there was no obvious difference in the surface morphology between the two. 4.3.3 Weight Difference Limit Inspection The weight difference limit requirements for the film formulation are shown in Table 4-3. The average weight range of ginseng oral film formulation is 0.02g to 0.2g, and the difference limit range must be controlled within 10%. The results of the weight difference limit test are shown in Table 4-4. 20 tablets of ginseng oral film were tested and measured, and the weight difference limit of the ginseng oral film was within 10%. The results show that the weight difference limit of the ginseng oral film meets the requirements. [Table 26] [Table 27] 4.3.4 In vitro residence time studies The release of Rg1 in ginseng extract requires that the ginseng oral film remain in the oral cavity for a long time, and that the drug penetrates the buccal mucosa and enters the blood circulation. Here, the in vitro residence time of the ginseng oral film was significantly extended (p<0.05), and in the experiment, it was observed that the erosion of the film by the solution started from the periphery of the film and eroded toward the center, and due to its own in vitro adhesiveness, the drug-loaded film could remain in the oral cavity for more than 3h under conditions simulating the washing away of saliva. The results show that the addition of a backing layer can effectively prevent the erosion of the film by the solution, extend the in vitro residence time, and ensure the release and penetration of Rg1. 4.3.5 Tensile strength test The stress curve and tensile strength results of the oral film are shown in Table 4-5. The results show that the addition of ginseng extract reduces the maximum resistance of the film (p<0.01) and the tensile strength is highly significantly reduced (p<0.001). The breaking elongation of the ginseng oral film is significantly increased (p<0.01), and the tensile strength test results show that the original uniform structure of the blank film is destroyed after it is loaded with the drug. [Table 28] Compared with blank oral film, *p<0.05, **p<0.01, ***p<0.001. 4.3.6 Toughness inspection The toughness test results are shown in Tables 4-6. The two films show that they can resist the surface compression force of the films to some extent, but the puncture resistance of the drug-loaded films was significantly decreased (p<0.05). Both the puncture and tensile experiments showed that the oral films destroyed their own structure after loading the drugs, reflecting the difference in mechanical properties. [Table 29] Note: * Compared with the puncture force of the blank film, p<0.05. 4.3.7 Wettability inspection The results of the wettability test are shown in Tables 4-7. When the wetting angle of the blank oral film and the ginseng oral film is less than 90°, it indicates that the surface has hydrophilicity and good wettability, which is favorable for the release of Rg1 in the film. Here, the wetting angle of the ginseng oral film is significantly increased compared with that of the blank oral film (p<0.05), which indicates that the addition of the extract increases the hydrophobicity of the surface, which may be related to the extraction method of the ginseng extract. The ginseng was extracted with 70% ethanol, and as a result, the extract had a certain hydrophobicity and the wetting angle increased. [Table 30] Note: * p<0.05 compared to the wetting angle of the blank film. 4.3.8 Content determination The content of Rg1 in the ginseng oral film is shown in Table 4-8. RSD<5% indicates that the content of Rg1 in the oral film is uniform. [Table 31] 4.3.9 Dissolution rate The dissolution rate results are shown in Figure 2. The cumulative release of the ginseng oral film in the artificial saliva dissolution medium was more than 80%, and the cumulative dissolution rate at 60 min was about 88.57%, indicating a good dissolution rate. 4.3.10 Stability-influencing factors testing (1) High temperature test The results of the high temperature test are shown in Table 4-9. Under the high temperature condition of 60°C, the film bends and loses weight, which is due to the film losing moisture under high temperature conditions, and the content of Rg1 has no difference compared to 0d. Therefore, the results of the high temperature test indicate that the film needs to be stored at room temperature or low temperature. [Table 32] (2) High humidity test The results of the high humidity test are shown in Table 4-10. Under the conditions of a temperature of 25°C and a humidity of 90%±5%, the film was seriously deformed, the four sides of the film were shrunk, and adhesion occurred between the oral films at 10d, and the film became soft. The content of Rg1 was not different from that at 0d. The results of the high humidity test show that the appearance of the film changed significantly under high humidity conditions, which is unfavorable for the storage of the film and may cause deterioration of the film in a humid environment, so the film should be stored in a dry place in an airtight container. [Table 33] (3) Light irradiation test The results of the light irradiation test are shown in Table 4-11. Under strong light conditions of 4500lx±500lx, the shape of the film changed, both ends were warped toward the middle, the weight was reduced, and the Rg1 content was not different from that of 0d. Therefore, the results of the light irradiation test indicate that the film needs to be stored in a dark place. [Table 34]

[0070] 4.4 Summary The appearance of the produced ginseng oral film meets the standard weight difference limit, the content is uniform, and when observed by scanning electron microscope, the film is smooth and flat, without drug crystals. It can be retained for more than 3 hours under the simulated in vitro oral environment, has good mechanical properties, and is easy to cut, use and transport. The dissolution rate reaches more than 80% within 1h, and it should be stored in a shady and dry place.

[0071] Test Example 5: Pharmacokinetics of ginseng oral film formulation 5.1 Materials and Equipment 5.1.1 Experimental animals Twelve golden hamsters, male, weighing 110±10g, were purchased from Changchun Changsheng Biological Technology Co., Ltd. License number: NO.SCXK(Liao)2020-0001, and were adapted and reared for one week in the animal room of our school at a temperature of (22±2)℃, ventilation, and humidity of (50±10)%. 5.1.2 Materials and Equipment [Table 35] [Table 36]

[0072] 5.2 Experimental Method 5.2.1 Preparation of standard solutions Preparation of Rg1 standard solution: 1.0 mg of Rg1 standard was precisely weighed, dissolved in methanol, and the volume was adjusted to a constant volume in a 10 mL measuring flask to prepare a standard stock solution with a concentration of 100 μg / mL. The solution was diluted to the desired concentration when used. Preparation of Sankinoside R1 standard substance as internal standard substance (IS): 1.0 mg of Sankinoside R1 was precisely weighed, dissolved in methanol and volumetrically adjusted in a 10 mL measuring flask to prepare Sankinoside R1 stock solution, which was then diluted to 5 μg / mL with methanol immediately before use. 5.2.1 Chromatographic and mass spectrometric conditions The chromatography and mass spectrometry conditions are shown in Tables 5-3 and 5-4. [Table 37] [Table 38] 5.2.3 Plasma sample processing 100μL of plasma sample was taken, 20μL of IS solution (R15μg / mL) was added, and mixed uniformly. Then, 1000μL of methanol, which is the extraction solvent, was added, vortexed for 10min to mix uniformly, and centrifuged for 15min (15000rpm). All the supernatant was taken and dried with a nitrogen blower. 200μL of methanol was added to redissolve, and vortexed for 5min to mix uniformly. All the solution was taken, filtered with a 0.22μm filter membrane, and the sample was injected and analyzed. 5.2.4 Methodological considerations (1) Consideration of linear relationships 50μL of blank plasma was taken, and 50μL of mixed standard solution with different concentrations was added to make the concentrations 0.1, 0.5, 1, 5, 10, 50, 100, 500, 1000ng / mL, respectively, and mixed uniformly. Then, the plasma was processed according to the plasma processing method in Section "2.1.3". The ratio of Rg1 peak area to R1 peak area was defined as Y, and the sample concentration was defined as X, and linear regression was performed. (2) Stability considerations Blank plasma samples were taken and quality control plasma samples containing Rg1 at concentrations of 5, 10, and 500 ng / mL were prepared. Plasma samples were left at room temperature for 24 h to evaluate short-term stability, stored in the autosampler for 24 h to evaluate autosampler stability, frozen and thawed at -80°C three times to evaluate freeze-thaw cycle stability, and frozen at -80°C for 7 d to evaluate long-term stability of the samples. (3) Consideration of precision and accuracy Blank plasma was taken, and three concentration samples, low, medium and high, were prepared and processed according to the plasma sample processing method in Section "2.1.3", and then injected into UHPLC-MS / MS for analysis. Three analytical batches were considered consecutively, and six samples were prepared for each concentration in each batch, and the peak areas of Rg1 and IS were recorded, and the intra-day and inter-day RSD values ​​of Rg1 and IS were calculated. (4) Consideration of matrix effects Three concentrations of Rg1 standard solution (5, 10, 500ng / mL) were prepared, treated according to the method in "2.1.3", and measured according to the conditions in "2.1.2". Six samples of each concentration were prepared in parallel, and the peak area 1 of Rg1 and IS was recorded. The supernatant of blank plasma was taken and prepared in three similar concentrations (5, 10, 500ng / mL), treated according to the method in "2.1.3", and measured according to the conditions in "2.1.2". Six samples of each concentration were prepared in parallel, and the peak area 2 of Rg1 and IS was recorded. The matrix effect of Rg1 and IS was calculated using the ratio of peak area 2 to peak area 1. (5) Consideration of recovery rate Blank plasma was taken and treated according to the plasma sample treatment method in section "2.1.3" under the condition of not adding IS, and then dried by blowing with nitrogen, and the corresponding concentrations of Rg1 and internal standard solution were added to make the concentrations 5, 10, and 500 ng / mL, and six samples of each were prepared in parallel, and the peak areas 3 of Rg1 and IS were recorded. The ratio of peak area 3 to peak area 2 in "2.1.4.4" was used to calculate the recovery rate. 5.2.3 In vivo pharmacokinetic studies in golden hamsters During the 7-day adaptation period, the golden hamsters' cheek pouches were washed with a cotton swab soaked in saline every day to keep them clean. Twelve healthy male golden hamsters weighing 100-120g were randomly divided into two groups: an intragastric administration group and an oral buccal administration group. Before the experiment, the golden hamsters were fasted for 24 hours, and their cheek pouches were washed with a cotton swab soaked in saline. The golden hamsters in the two groups were administered 500mg / kg of ginseng extract and an equal amount of ginseng oral film (which was placed in the left and right cheek pouches of the golden hamsters at the same time). 0, 0.08, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 5, and 6 hours after administration, 0.3mL of blood was collected from the orbital venous plexus and placed in a tube moistened with heparin sodium saline. The collected whole blood was centrifuged (15,000 rpm, 10 min), 100 μL of the supernatant was taken, treated according to "2.1.3", measured according to the conditions in "2.1.2", and the data was recorded. 5.2.4 Data Processing The main pharmacokinetic parameters were calculated according to the one-compartment model method using DAS 2.0 software and are shown in Tables 5-8. The calculation formula for relative bioavailability is shown in Equation 7.

number

[0073] 5.3 Experimental results 5.3.1 Appearance of the golden hamster cheek pouch When the cheek pouch of a golden hamster was opened, abundant capillaries and a large area of ​​buccal mucosa were observed. After the ginseng oral film formulation adhered to the buccal mucosa, it could not be expelled, which makes it a very suitable pharmacokinetic model for the oral film formulation. 5.3.2 Results of consideration of UHPLC-MS / MS methodology (1) Calibration curve The ion chromatograms of Rg1 and R1 in golden hamster plasma detected by UHPLC-MS / MS showed that Rg1 and R1 had sharp chromatographic peak shapes under the chromatographic and mass spectrometry conditions in section "2.1.2". At the same time, endogenous substances or metabolites in plasma did not interfere with the measurement of the target components, and the retention times of Rg1 and R1 were 3.81 min and 3.65 min, respectively. The regression equation was Y=770.44X-144.05, and Rg1 in golden hamster plasma had a good linear relationship within the range of 0.1-1000 ng / mL, and R 2 =0.9999. (2) Stability considerations Stability considerations and experimental results are shown in Table 5-5. The results show that the stability of plasma samples in the four processing methods meets the relevant pharmacopoeia regulations of RSD<10%, and the plasma sample processing methods and UHPLC-MS / MS detection methods are reasonable. [Table 39] (3) Precision Considerations Precision considerations and experimental results are shown in Table 5-6. The intraday and interday precision of Rg1 are both less than 3%, which meets the Pharmacopoeia requirements. [Table 40] (4) Recovery rate and matrix effect The matrix effect observation results are shown in Tables 5-7. From the tables, the recovery rate of the method is >80%, and the deviation between the matrix effect and the recovery rate is less than 8%, indicating that the sample processing method is stable and reliable. [Table 41] 5.3.3 Average blood concentration-time curve After the same dose of ginseng extract (500 mg / kg) was administered intragastrically and orally to golden hamsters, the mean blood concentration-time curve of Rg1 in plasma is shown in Figure 3. Compared with the intragastric administration group, the area under the Rg1 blood concentration-time curve AUC in the orally administered group was significantly increased. In the case of intragastric administration, a second drug peak appeared at about 1.5 h, which was due to the fact that interconversion occurred between ginsenosides in the acidic environment of the golden hamster's gastrointestinal tract, increasing the content of Rg1 in plasma. The specific reason for the appearance of the second drug peak requires further study. 5.3.4 In vivo pharmacokinetic parameter analysis in golden hamsters The main kinetic parameters were calculated from the measured data, and the results are shown in Tables 5-8. AUC of the measured components of Rg1 in plasma after administration of the same dose of ginseng extract (0-t) The intragastric administration group had a significantly lower half-life than the orobuccal administration group (p<0.001). Compared with the intragastric administration group, the orobuccal administration group significantly prolonged the drug half-life of Rg1 in golden hamsters (p<0.05). However, C max There was no significant difference in AUC (0-t) showed that the oral buccal administration group was higher than the intragastric administration group, because the ginseng oral film could adhere to the cheek pouch of golden hamsters for about 3 h and enter the systemic blood circulation through the buccal mucosa. The relative bioavailability F of oral buccal administration was 369.57%, indicating that the preparation of the ginseng oral film achieved the formulation purpose. [Table 42] Compared with the intragastric administration group Rg1, * p<0.05, ** p<0.01, *** p<0.001.

[0074] 5.4 Summary After the ginseng extract was made into an oral film, it was attached to the surface of the buccal mucosa of golden hamsters for a long time, which significantly extended the half-life of Rg1 in the blood. Compared with the intragastric administration method, the AUC (0-t) increased by 3.70 times, and the relative bioavailability was 369.57%, and the results showed that the ginseng oral film could achieve the formulation purpose and effectively improve the bioavailability.

[0075] As can be seen from the above examples, the use of the ginseng extract provided by the present invention in the production of film preparations can effectively extract ginsenoside Rg1 by reflux extraction using an aqueous ethanol solution as the extraction solvent, and by limiting the appropriate parameter range so that the content of ginsenoside Rg1 in the obtained ginseng extract is high, it can reduce raw material costs and is suitable for mass production.

[0076] The ginseng oral film provided by the present invention can directly act on the oral buccal mucosa, which has a weaker barrier function than the skin, and can avoid the inhibitory effect of the skin stratum corneum on the penetration of ginsenoside and the first-pass effect, and can also play a role in improving the bioavailability of Rg1. At the same time, the ginseng oral film is smooth and flat, free of drug crystals, has good mechanical properties, good dissolution rate, is easy to take, and has good patient compliance.

[0077] Although the above embodiments have described the present invention in detail, they are only some of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained based on the embodiments without inventive step, and all of these embodiments belong to the protection scope of the present invention.

Claims

1. A use of a ginseng extract in the production of a film preparation, the production method of the ginseng extract comprising: The method includes a step of reflux-extracting ginseng using an aqueous ethanol solution as an extraction solvent to obtain the ginseng extract, The mass ratio of the ginseng to the aqueous ethanol solution is 0.5 to 1:6 to 8; The volume fraction of ethanol in the ethanol aqueous solution is 65 to 75%, The use of a ginseng extract in the production of a film preparation, wherein the extraction time for a single reflux extraction is 0.5 to 1.5 hours and the number of reflux extractions is 1 to 3.

2. A ginseng oral film formulation comprising a backing layer and a drug-containing layer laminated in order, the drug-containing layer comprises, by weight, 10 to 12 parts of hydroxypropyl methylcellulose, 0.1 to 1 part of carbomer, 9 to 11 parts of ginseng extract, and 0.1 to 0.3 parts of fatty acid, the ginseng extract being the ginseng extract according to claim 1; The ginseng oral film preparation, wherein the backing layer contains 0.6 to 0.8 parts by mass of ethyl cellulose based on the parts by mass of the hydroxypropyl methyl cellulose.

3. The ginseng oral film preparation according to claim 2, wherein the fatty acid comprises at least one of caproic acid, caprylic acid, capric acid, lauric acid, stearic acid, oleic acid and linoleic acid.

4. sequentially performing a first injection of ethyl cellulose solution into an anti-adhesive container and a first molding to obtain a backing layer; swelling carbomer in water and first mixing with hydroxypropyl methylcellulose to obtain a first film solution; performing a second mixing of a fatty acid solution with the first film liquid to obtain a second film liquid; a third mixing step of mixing the ginseng extract with the second film liquid to obtain a drug-containing layer film liquid; and performing a second injection and a second molding of the drug-containing layer film liquid into a backing layer in sequence to obtain the ginseng oral film preparation.

5. The method according to claim 4, wherein the temperature of the first mixture is 70 to 90°C.

6. The density of the ethyl cellulose solution is 0.7 to 0.9 g / cm ３ and The density of the drug-containing layer film liquid is 1.3 to 1.4 g / cm ３ The method according to claim 4, characterized in that

7. The method according to claim 4, wherein the temperature of the first molding is 50 to 70° C. and the time is 20 to 40 minutes.

8. The method according to claim 4, wherein the temperature of the second molding is 60 to 80° C. and the time is 1 to 3 hours.

9. Use of the ginseng oral film preparation according to claim 2 or 3 in the manufacture of an anti-cancer preparation or an anti-diabetic preparation.

10. Use of the ginseng oral film preparation according to claim 2 or 3 in the manufacture of a blood pressure regulating preparation, a blood sugar regulating preparation or an antioxidant preparation.

Images