Use of Panax ginseng extract in the manufacture of film preparations, Panax ginseng oral film preparations and their manufacturing methods and uses

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] This invention belongs to the field of traditional Chinese medicine, and more specifically, relates to the use of ginseng extract in the production of film preparations, ginseng oral film preparations, and methods and uses thereof. [Background technology]

[0002] Panax ginseng (CAMey.), a perennial plant belonging to the Araliaceae family, is known to have various beneficial effects, including improving memory decline and spatial cognitive ability, as well as anti-cancer and anti-diabetic activity. Its functional benefits also include improving liver function, regulating blood pressure and blood glucose, and providing antioxidant activity. Panax ginseng contains various components such as ginsenosides, acidic polysaccharides, flavonoids, organic acids, polyacetylenes, alkaloids, and phenolic compounds. Among these, ginsenosides such as Rg1, Rb1, Re, Rg3, and Rh2 are the main pharmacologically active components of Panax ginseng, with Rg1 exhibiting more biological activity and attracting widespread attention.

[0003] Water decoction and ethanol reflux extraction are conventional extraction methods. These two methods have the advantages of being easy to operate and low-cost, and are still commonly used in many production companies and laboratories in actual production processes. However, the extraction rate of ginsenoside Rg1 is poor in current extraction processes. [Overview of the project] [Problems that the invention aims to solve]

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

[0005] To achieve the above object, the present invention provides the following technical solutions.

[0006] The present invention provides the use of the otane carrot extract in the manufacture of a film agent, and the method for manufacturing the otane carrot extract is as follows. Using an aqueous ethanol solution as an extraction solvent, reflux-extracting otane carrot, and obtaining the otane carrot extract, including the step of The mass ratio of the otane carrot 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 0.5-1.5 h, and the number of times is 1-3 times.

[0007] The present invention further provides an otane carrot oral film agent, including a backing layer and a drug-containing layer laminated in sequence, The drug-containing layer contains, in parts by mass, 10-12 parts of hydroxypropyl methylcellulose, 0.1-1 part of carbomer, 9-11 parts of otane carrot extract, and 0.1-0.3 part of fatty acid. The otane carrot extract is the otane carrot extract described in the above technical solution. The backing layer contains 0.6-0.8 parts of ethylcellulose based on the number of parts by mass of the hydroxypropyl methylcellulose.

[0008] Preferably, the fatty acid includes 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 manufacturing the otane carrot oral film agent described in the above technical solution. Sequentially performing a first injection and a first molding of an ethylcellulose solution into an anti-adhesion container to obtain a backing layer, ​​The steps include: performing a second mixing of the fatty acid solution and the first film solution to obtain a second film solution; The third step involves mixing the aforementioned ginseng extract with the second film solution to obtain a drug-containing layer film solution. The procedure includes the steps of sequentially injecting the drug-containing layer film liquid into the backing layer and performing a second molding to obtain the ginseng oral film preparation.

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

[0011] Preferably, the density of the ethylcellulose solution is 0.7 to 0.9 g / cm³. 3 And, The density of the drug-containing film solution is 1.3-1.4 g / cm³. 3 That is the case.

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

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

[0014] The present invention further provides the use of the ginseng oral film preparation described above in the manufacture of an anticancer drug or an antidiabetic drug.

[0015] The present invention further provides the use of the ginseng oral film preparation described above in the manufacture of blood pressure regulating preparations, blood glucose regulating preparations, or antioxidant preparations.

[0016] The present invention provides the use of ginseng extract in the production of a film agent, the method for producing the ginseng extract comprising the steps of using an aqueous ethanol solution as the extraction solvent, reflux extracting ginseng to obtain the ginseng extract, wherein the mass ratio of ginseng to aqueous ethanol solution is 0.5 to 1:6 to 8, the volume fraction of ethanol in the aqueous ethanol solution is 65 to 75%, the single reflux extraction time is 0.5 to 1.5 hours, and the number of reflux extractions is 1 to 3. The present invention uses an aqueous ethanol solution as the extraction solvent, effectively extracts ginsenoside Rg1 by reflux extraction, and reduces raw material costs and is suitable for mass production by limiting the parameter range appropriately so that the ginsenoside Rg1 content in the obtained ginseng extract is high. [Effects of the Invention]

[0017] The ginseng oral film formulation provided by the present invention acts directly on the oral buccal mucosa, which has a weaker barrier function than the skin, thereby avoiding the inhibitory effect on the penetration of ginsenosides into the stratum corneum of the skin and the first-pass effect, and can further play a role in improving the bioavailability of Rg1. At the same time, the ginseng oral film formulation is smooth and flat, free of drug crystals, has good mechanical properties, has good solubility, is easy to take, and has good patient compliance. [Brief explanation of the drawing]

[0018] To more clearly illustrate embodiments of the present invention or technical solutions of the prior art, the drawings necessary for use in the embodiments are briefly described below. Of course, the drawings described below are merely a part of embodiments of the present invention, and those skilled in the art can obtain other drawings based on these without any creative effort.

[0019] [Figure 1] This is the appearance of the ginseng oral film preparation described in Example 2. [Figure 2] This is the dissolution rate of the ginseng oral film preparation described in Example 2 in artificial saliva. [Figure 3] These are the plasma concentration-time curves for Rg1 administered intragastricly and orally and buccally. [Modes for carrying out the invention]

[0020] The present invention provides the use of ginseng extract in the production of a film agent, and the method for producing the ginseng extract is as follows: The process includes the step of using an aqueous ethanol solution as the extraction solvent, reflux extracting Panax ginseng, and obtaining the Panax ginseng extract. The mass ratio of the aforementioned ginseng to the ethanol aqueous solution is 0.5 to 1:6 to 8. The volume fraction of ethanol in the aforementioned aqueous ethanol solution is 65-75%. The single extraction time for the reflux extraction described above is 0.5 to 1.5 hours, and the number of extractions is 1 to 3.

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

[0022] In this invention, an aqueous ethanol solution is used as the extraction solvent, and Panax ginseng is reflux-extracted to obtain the Panax 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 65-75%, preferably 67-73%, more preferably 69-71%. The single extraction time for reflux extraction is 0.5 to 1.5 hours, preferably 0.7 to 1.3 hours, more preferably 0.9 to 1.1 hours, and the number of extractions is 1 to 3 times, more preferably 2 to 3 times.

[0024] The use of the ginseng extract provided in this invention in the production of film preparations effectively extracts ginsenoside Rg1 by reflux extraction using an aqueous ethanol solution as the extraction solvent. By limiting the parameter range appropriately so that the ginsenoside Rg1 content in the obtained ginseng extract is high, raw material costs are reduced, making it suitable for mass production.

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

[0026] The drug-containing layer comprises 10 to 12 parts by mass, 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 mass parts 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 mass parts of the hydroxypropyl methylcellulose.

[0029] The drug-containing layer contains 0.1 to 0.3 parts of fatty acids, preferably 0.2 to 0.3 parts, based on the mass parts of 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 one or more of caprylic acid, capric acid, oleic acid, and linoleic acid, and most preferably capric acid. If 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 ethylcellulose based on the mass parts of hydroxypropyl methylcellulose.

[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 formulation provided by the present invention acts directly on the oral buccal mucosa, which has a weaker barrier function than the skin, thereby avoiding the inhibitory effect on the penetration of ginsenosides into the stratum corneum of the skin and the first-pass effect, and can further play a role in improving the bioavailability of Rg1. At the same time, the ginseng oral film formulation is smooth and flat, free of drug crystals, has good mechanical properties, has good solubility, is easy to take, and has good patient compliance.

[0035] The present invention further provides a method for producing a ginseng oral film preparation as described in the above technical solution, The first step involves sequentially injecting an ethylcellulose solution into an anti-adhesion container and performing a first molding to obtain a backing layer. The steps include swelling carbomer in water, mixing it with hydroxypropyl methylcellulose to obtain a first film solution, and The steps include: performing a second mixing of the fatty acid solution and the first film solution to obtain a second film solution; The third step involves mixing the aforementioned ginseng extract with the second film solution to obtain a drug-containing layer film solution. The procedure includes the steps of sequentially injecting the drug-containing layer film liquid into the backing layer and performing a second molding to obtain the ginseng oral film preparation.

[0036] In this invention, an ethylcellulose solution is sequentially injected into an anti-adhesion container and molded to obtain a backing layer.

[0037] In the present invention, the concentration of the ethylcellulose 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, and the density of the ethylcellulose solution is preferably 0.7 to 0.9 g / cm³. 3 , more preferably 0.75~0.85 g / cm³ 3 Most preferably 0.80-0.83 g / cm³ 3 The solvent for the ethylcellulose solution is preferably ethanol, more preferably anhydrous 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 min, more preferably 25 to 35 min, and most preferably 30 to 35 min, and the first molding is preferably carried out by vacuum drying.

[0039] In this invention, after obtaining the backing layer, the carbomer is swollen in water and mixed with hydroxypropyl methylcellulose to obtain a first film solution.

[0040] In the present invention, the mass ratio of 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 this invention, after obtaining a first film solution, a second mixing is performed between the fatty acid solution and the first film solution to obtain a second film solution.

[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 of the fatty acid solution is preferably propylene glycol.

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

[0045] In this invention, after obtaining the second film solution, a third mixing is performed between the ginseng extract and the second film solution to obtain a drug-containing layer film solution.

[0046] In the present invention, the density of the drug-containing layer film liquid is preferably 1.3 to 1.4 g / cm³. 3 , more preferably 1.31 to 1.35 g / cm³ 3 Most preferably 1.31-1.32 g / cm³ 3 And, In the present invention, the third mixing is preferably carried out at room temperature, and the present invention does not impose any special limitations on the process of the third mixing, and any method well known to those skilled in the art may be used.

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

[0048] In the present invention, the temperature of the second molding 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 h, more preferably 1.5 to 2.5 h, and most preferably 2 to 2.5 h. The second molding is preferably carried out by a vacuum drying method.

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

[0050] The present invention further provides the use of the otane ninjin oral film agent described in the above technical solution in the manufacture of an anticancer agent or an antidiabetic agent.

[0051] The present invention further provides the use of the otane ninjin oral film agent described in the above technical solution in the manufacture of a blood pressure regulating agent, a blood glucose regulating agent or an antioxidant agent.

[0052] To further illustrate the present invention, the use of the film agent of the otane ninjin extract provided by the present invention, the otane ninjin oral film agent and its manufacturing method and use will be described in detail below with reference to the drawings and examples, but they should not be understood as limiting the protection scope of the present invention.

[0053] Example 1 Using an ethanol aqueous solution with a volume fraction of 70% as the extraction solvent, with a solid-liquid ratio of 1:8, an extraction time of 1 h, and an extraction number of 2 times, reflux extraction was performed on the otane ninjin extract to obtain the otane ninjin 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 it was poured into a petri dish at a density of 0.81 g / cm 3 and vacuum dried at 60°C for 30 min to obtain a backing layer. 0.786 g of carbomer was swollen in water for 12 hours, 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 hour to obtain the 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 the second film solution. After the second film solution cooled to room temperature, 10.155 g of ginseng extract was added and stirred until dissolved, resulting in a density of 1.31 g / cm³. 3 A drug-containing layer film solution was obtained. A drug-containing film solution was injected into the backing layer and vacuum-dried 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 Instruments 1.1.1 Experimental materials and reagents Table 1-1 shows the materials and reagents used in this experiment. [Table 1] 1.1.2 Laboratory Equipment Table 1-2 shows the experimental equipment used in this experiment. [Table 2]

[0056] 1.2 Method for measuring the HPLC content of Rg1 The Rg1 content in the extract was measured using high-performance liquid chromatography. 1.2.1 Chromatography Conditions Table 1-3 shows the chromatography conditions for Rg1 measurement by HPLC. [Table 3] 1.2.2 Preparation of Solutions Standard substance solution: 5.0 mg of Rg1 standard substance was weighed, methanol was added to bring the volume to 10 mL, and a Rg1 0.5 mg / mL solution was prepared. This solution was then diluted to the appropriate concentration at the time of use. Test substance solution: 100.0 mg of dried ginseng extract was weighed, placed in a 10 mL volumetric flask, and 5 mL of methanol was added. The solution was sonicated for 30 minutes, allowed to cool, and methanol was added to the mark to obtain 10 mL of the test substance solution. 1.2.3 Results of the specificity experiment The specificity result shows that the retention time for Rg1 is 8.10 min, and the chromatographic peak shape of this component is good, and it is well separated from the solvent peak and other components. Therefore, these chromatographic conditions can be used to measure the Rg1 content. 1.2.4 Results of methodological considerations Tables 1-4 and 1-5 show the methodological considerations for measuring the Rg1 content. The results indicate that the established HPLC method is reliable and can be used to measure the Rg1 content in ginseng extract. [Table 4] [Table 5]

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

[0058] 1.4 Summary Water decoction and ethanol reflux extraction are conventional extraction methods. These two methods have the advantages of being easy to operate and low-cost, and are still commonly used in many production companies and laboratories in actual production processes. To increase the extraction rate of Rg1, it is necessary to select the optimal extraction process suitable for actual production. The results of orthogonal design tests of the water decoction method showed that the extraction rate of Rg1 decreased with increasing extraction time and number of extractions. On the one hand, prolonged heating may decompose Rg1, while on the other hand, compounds such as ginseng polysaccharides, which have higher solubility in water, have a higher extraction rate in water. In summary, the optimal extraction process for producing ginseng extract is to use an aqueous ethanol solution as the extraction solvent, reflux extract the ginseng, and 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 aqueous ethanol solution is 65 to 75%, the single reflux extraction time is 0.5 to 1.5 hours, and the number of extractions is 1 to 3.

[0059] Study Example 2: Study on the permeability of fatty acids in ginseng extract This study primarily investigated the osmotic effect of fatty acids on Rg1, using Rg1 in ginseng extract as an indicator component, and five saturated fatty acids, namely caproic acid (C6), caprylic acid (C8), and capric acid (C8). 10 ), lauric acid (C 12 ), stearic acid (C 18 ), two types of unsaturated fatty acids, namely oleic acid (C 18:1 ), linoleic acid (C 18:2 The penetration-enhancing effect of [substance name] is examined, with the indicators considered including steady-state flux, apparent permeability coefficient, permeability enhancement factor, delay time, and 4-hour cumulative permeation. The permeability-enhancing effect of different fatty acids is evaluated using principal component analysis, providing a reference basis for the rational use of permeability enhancers in ginseng oral film preparations.

[0060] 2.1 Reagents and Instruments 2.1.1 Experimental materials and reagents Table 2-1 shows the materials and reagents used in this experiment. [Table 10] 2.1.2 Laboratory Equipment Table 2-2 shows the experimental equipment used in this experiment. [Table 11]

[0061] 2.2 Method 2.2.1 Preparation of Solutions 110.0 mg of Rg1 standard substance was weighed, dissolved in 10 mL of PBS solution at pH=6.8, shaken for 1 hour, and sonicated for 1 hour to prepare 10 mL of ginseng extract feed solution containing the same concentration (11 mg / mL) of Rg1. The feed solution was stored at 4°C in preparation for use, and the feed solution was vortexed for 5 minutes before use. 2.2.2 Preparation of fatty acid penetration 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 osmotic accelerating solutions at concentrations of 1 to 10 mg / mL for each. 2.2.3 In vitro osmosis experiment The treated porcine esophageal epithelium was placed in a Franz diffusion cell (diffusion area 1.76 cm²). 2 The porcine esophageal epithelium was placed in the center of the receiving chamber (4.5 mL of solution volume). 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. 1.0 mL of pH=6.8 PBS solution was added to the supply chamber, and after 5 minutes, it was checked whether the solution in the supply chamber leaked into the receiving chamber due to damage to the porcine esophageal epithelium or improper installation of the diffusion cell. After confirming that there were no abnormalities, the solution in the supply chamber was discarded and wiped clean. 40.0 μL of penetration enhancer solution was added to the supply chamber, and the supply chamber was left open for 1 hour to ensure evaporation and drying of the ethanol solvent and eliminate solvent interference. 4.5 mL of pH=7.4 PBS solution was added to the receiving chamber, 0.2 mL of supply solution was added to the supply chamber, the Franz diffusion cell was placed in the transdermal diffusion apparatus, a magnetic stirring bar was added, the rotation speed was set to 600 rpm, and the temperature was set to 37°C ± 0.5°C. At 0, 0.25, 0.5, 0.75, 1, 1.5, 2.5, 3, and 4 hours, 0.3 mL of the receiving solution was aspirated from the receiving chamber, and 0.3 mL of fresh isothermal pH=7.4 PBS buffer was added to the receiving chamber. 2.2.4 Data Analysis The measured porcine esophageal epithelial infiltration data consisted of the apparent osmotic coefficient P, delay time TL, and steady-state flow rate J, and the calculation methods are shown in Equations 1, 2, 3, and 4.

number

number

number

number

[0062] 2.3 Results 2.3.1 Penetration-promoting effect of seven types of fatty acids on Rg1 During the osmosis experiment, the concentration of the recipient solution was able to reach sink conditions at all times, and the drug concentration of the recipient solution was measured at different time points. Calculations were performed on the data to obtain parameters such as the steady-state flux of Rg1 osmosis through the porcine esophageal epithelium, apparent osmosis coefficient, osmosis enhancement factor, 4-hour cumulative permeation, and delay time, which are shown in Table 2-3. The results indicate that the cumulative permeation of Rg1 has a good linear relationship with time. Compared to the blank group, 1 mg / mL and 5 mg / mL stearic acid (C) showed a significant difference. 18 ) showed no significant difference in ER (p>0.05), proving that 1 mg / mL and 5 mg / mL stearic acid had no osmotic effect on Rg1. All remaining fatty acids could increase the osmotic coefficient of Rg1 to varying degrees, and among saturated fatty acids, 1 mg / mL capric acid (C) 10 The osmotic enhancement factor was highest for 5 mg / mL of linoleic acid (C15). ER: 19.24±1.89. Among unsaturated fatty acids, the one with the highest osmotic enhancement factor was linoleic acid (C15) at 5 mg / mL.18:2 ) 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) Experiment on the applicability of standardization processing and factor analysis of penetration data The data in Table 2-3 was input into IBM SPSS Statistics 26.0 software, standardized to eliminate the influence of units in different indicators, and standardized data was obtained. Next, an applicability experiment for factor analysis was conducted, and the results are shown in Table 2-4. When conducting information enrichment research using principal component analysis, it is first necessary to analyze whether the data is suitable for principal component analysis. The KMO value is equal to 0.797, which indicates that there is a certain relationship between the five indicators. In the Bartelett sphericity test, the Sig value is 0.000 (p<0.05), which rejects the identity matrix for the correlation coefficient, meaning that each indicator is related. Both result tests indicate that the data is suitable for principal component analysis. [Table 13] (2) Calculation of principal component feature vectors The eigenvalues ​​and variance contribution rates of the correlation coefficients were calculated using 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%, indicating that Component 1 and Component 2 can represent 99.78% of the five indicator information. Based on this, it can be concluded that Component 1 and Component 2 can represent a comprehensive judgment of the penetration-promoting 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 × 4h cumulative permeation + 0.085 × delay time; Component 2 = -0.085 × steady-state flux -0.085 × apparent permeability coefficient -0.085 × permeability enhancement factor -0.107 × 4h cumulative permeation + 1.035 × delay time. (3) Data calculation for principal component scores and total scores The standardized data for each processed variable is substituted into a linear equation, and the total score is calculated cumulatively by multiplying the explanatory power of variance by the component scores. The formula for the total score is: Total score = (81.66% × Component 1 + 18.12% × Component 2) / 99.783%. The total scores are sorted, and the results are shown in Table 2-6. From the ranking in the table, it was found that 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. From the overall score, it was found that 1% capric acid had the highest penetration-promoting effect. [Table 15]

[0063] 2.4 Summary Fatty acids can enhance the permeability of Rg1 through the porcine esophageal epithelium, reaching 0.227 mg / cm³. 2 When capric acid acts on the esophageal epithelium of pigs, it has the greatest penetration-enhancing effect.

[0064] Study Example 3: Preparation of a Ginseng-Based Oral Film Formulation 3.1 Reagents and Equipment 3.1.1 Experimental materials and reagents Table 3-1 shows the materials and reagents used in this experiment. [Table 16] 3.1.2 Laboratory Equipment Table 3-2 shows the experimental equipment used in this experiment. [Table 17]

[0065] 3.2 Method The ginseng oral film preparation is manufactured as a two-layer film preparation. One layer is a water-insoluble backing layer, which not only controls the unidirectional release of the drug but also protects the drug-containing layer from erosion by saliva. The other layer is the drug-containing layer, which is made of a hydrophilic polymer material with a certain in vitro adhesive strength, ensuring that the film preparation adheres to the surface of the buccal mucosa and maintaining drug penetration. 3.2.1 Selection of Film Forming Method Common production methods for film formulations include coating, solvent injection, composite film formation, hot melt extrusion, thermoplastic film formation, semi-solid solvent injection, solid dispersion extrusion, and spheroidization. In production, coating and solvent injection are generally used. These two film formation methods are easy to operate, dry quickly, and have high production rates. However, coating is unsuitable for the production of herbal medicine films due to its difficulty in controlling thickness and low drug load capacity. Solvent injection allows for control of the amount and area of ​​film-forming material used by pouring an adhesive solution into a container of a fixed volume. This method is suitable for manufacturing herbal medicine films in the laboratory. 3.2.2 Selection of drug-containing layer film forming material Film agents are essentially polymer matrices, composed of one or more polymers. Currently, commonly used materials include gelatin, shellac, carboxymethylcellulose, pullulan, hydroxypropylcellulose, hydroxypropylmethylcellulose, and sodium alginate. Due to its excellent mechanical properties and drug-carrying ability after film formation, hydroxypropylmethylcellulose was selected as the main film-forming material in this study. Furthermore, because cellulose derivatives alone have low mucosal adhesion, an appropriate amount of carbomer was added as an adhesive to improve the bioadhesion of the drug carrier. At the same time, the combination of carbomer and hydroxypropylmethylcellulose can maintain drug release as a sustained-release skeletal material for the film agent. 3.2.3 Selection of Plasticizer for Drug-Containing Layer Film agents manufactured using film-forming materials alone have a hard texture and poor mechanical properties. The addition of plasticizers improves the flexibility of the film agent and reduces its brittleness, which is advantageous for the transport and storage of the film agent. When glycerin is used as a plasticizer, adhesion occurs between the film agents, which is unfavorable for storage. When triethyl citrate is used as a plasticizer, the surface of the film agent is smooth, but it is highly brittle and has low flexibility. When propylene glycol is used as the plasticizer in this formulation, the surface of the film agent is smooth, and the flexibility is appropriate, making it suitable as the plasticizer in this formulation. 3.2.4 Selection of backing layer material While ginseng is readily soluble in water as the main ingredient, and its extract can be uniformly dispersed in the film formulation, its bitter taste reduces patient compliance, making oral film formulation difficult to administer. Simultaneously, single-layer drug-containing films are eroded by saliva, rapidly swelling and disintegrating, and cannot remain in the oral cavity for extended periods. Considering patient compliance and oral retention time, a two-layer oral film formulation is created by adding a backing layer to the drug-containing layer. The backing layer requires both that the selected film-forming material is insoluble in water at human body temperature and adheres tightly to the drug-containing layer. Ultimately, ethylcellulose is selected as the backing layer material. 3.2.5 Consideration method (1) Appearance of the film The requirements for the appearance of the film are that it is complete and smooth, has consistent thickness, uniform color, and is free of obvious air bubbles. In the case of multi-dose film, the segmentation indentations should be uniform and clear, and the film should be able to be torn by pressing on the indentations. (2) Film solution Although not required by the pharmacopoeia for film solutions, the solvent injection method requires that the film solution have appropriate fluidity. If the film solution is too viscous, it is difficult to remove air bubbles and pour it out of the container. If the film solution is too fluid, the drying time is long and it is difficult to control the quality of the manufactured film. The adhesive liquid system should be uniform, and the standard is that there should be no obvious insoluble lumps or solids visible to the naked eye. (3) Dissolution time A (2.2cm × 3.2cm) oral film was placed on top of a petri dish, and 200 μL of 37°C artificial saliva (KH2PO4 12 mM, NaCl 40 mM, CaCl 21.5 mM, H2O 1 L, pH adjusted to 6.8) was dropped onto the film. This process continued until the droplet penetrated the film and dripped into the petri dish. The time it took for the droplet to penetrate the film and drip to the bottom of the petri dish was measured and recorded as the dissolution time. Six measurements were taken for each group of samples, and the average value was recorded. (4) In vitro adhesive strength In this specification, the administration site for the oral film agent is the oral buccal mucosa. This requires the oral film agent to adhere to the oral buccal mucosal tissue, release Rg1, penetrate the buccal mucosa, and enter the bloodstream. Therefore, the film agent must have a certain degree of adhesion to the oral mucosa. Using an Eidburg HLB universal material tester, one slide glass was fixed to the base. Using another slide glass, a clip was fixed to the center point of the slide glass, and the prepared slide glass with the clip attached was fixed to the upper clip of the universal material tester, so that the two slide glasses were horizontal. Next, the manufactured film isolation layer was fixed to the upper slide glass with a clip using double-sided tape. 200 μL of 5% (w / w) mucin solution was dropped onto the base slide glass, bringing the upper and lower slide glasses into close contact. A 2g weight was placed on top for 30 seconds, and then the slide glass was pulled upward at a constant speed of 0.1 mm / s. At the moment the two parts of the slide glass were separated, the tension reading displayed on the universal material tester at that time was recorded as the in vitro adhesive strength of the film material.

[0066] 3.3 Results 3.3.1 Optimization of the formulation composition of blank film agents using the Bok-Behnken response surface method 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 on three selected factors, with the dissolution time of the blank film agent and in vitro adhesive strength as evaluation indices (Y1) and (Y2). Contour plots and three-dimensional response plane diagrams were created using the software for two indices and three factors. [Table 18] Using Design Expert 12 software, statistical analysis was performed on the data in the table, and the multiple nonlinear fitting equations and related coefficients for which dissolution time fits each factor are 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 a statistically significant difference, indicating that the nonlinear model used is significant. At the same time, if the value of the misfit term is greater than 0.05, it is not significant, indicating that the equation fits the experiment well. Here, B, B 2 , C 2 This has a significant impact on Y1. [Table 19] The multiple nonlinear fitting equations and related coefficients that fit each factor with in vitro adhesive force are 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 2R=0.9723. The results of the analysis of variance are shown in Table 3-5. p<0.05 is considered statistically significant, indicating that the nonlinear model used is significant. At the same time, if the value of the misfit term is greater than 0.05, it is not significant, indicating that the equation fits the experiment well. Here, A, B, C, B 2 This has a significant impact on Y2. [Table 20] Next, using the analysis module of Design Expert 10.0 software, we created three-dimensional effect area diagrams and contour plots for the total score of the interaction of each factor. We found that the amount of carbomer used (A), HPMC used (B), and propylene glycol used (C) had some effect on the dissolution time and in vitro adhesion of the blank film agent. After processing the experimental results with Design-Expert 12 software, we used the longest dissolution time and maximum in vitro adhesion as evaluation indices to obtain an optimized film agent formulation, namely, carbomer: 0.1-1g, HPMC: 1-5g, and propylene glycol: 0.1-0.3g. Three validation tests were conducted according to the optimized formulation. The average dissolution time obtained was 225.58 s, and the average in vitro adhesive strength was 9.63 N. The relative errors from the predicted values ​​were 0.48% and 0.31%, respectively, demonstrating that the formulation is stable and reliable. 3.3.2 Consideration of drug load When the ginseng extract accounted for 25%, 45%, and 65% of the solid content, respectively, microscopic observation revealed that when the ginseng extract accounted for 65% of the solid content, highlight crystals appeared in the field of view. Ultimately, it was determined that the drug-carrying amount was 45% of the solid content. 3.3.3 Selection of drying equipment We investigated drying methods for producing ginseng oral film preparations according to an optimized formulation. Three drying methods were selected based on existing laboratory conditions: vacuum drying, hot air drying box drying, and heated plate drying. These were then selected based on the appearance evaluation index of the pharmacopoeia, and the experimental results are shown in Table 3-6. Vacuum drying utilizes the principle of vacuum negative pressure to lower the boiling point of water, accelerating the evaporation of water in the film solution. After drying, the film surface is smooth, flat, and uniform, and free of air bubbles. Hot air drying box drying involves filling the drying box with flowing hot air to remove water from the film solution. After drying, the hot air blows onto the surface of the film solution, causing wrinkles and making the film uneven, thus failing to meet the appearance requirements of the pharmacopoeia. Heated plate drying results in uneven heating of the film solution due to the uneven placement of heating devices in the equipment and the placement of a petri dish between the heated plate and the film solution, leading to varying film thicknesses. Therefore, vacuum drying was selected as the drying method for the film solution. [Table 21] 3.3.4 Selection of drying temperature and time Table 3-7 shows the results of the selection of drying temperature and time. Vacuum drying was selected as the drying method, and the minimum moisture content of the ginseng oral film was used as the selection criterion for drying temperature and drying time. When the temperature was 70°C, there was no significant difference in the data between 2h and 3h (p>0.05), and ultimately, a drying temperature of 70°C and a drying time of 2h were selected to save drying time and cost. [Table 22] 3.3.5 Optimal Formulation Table 3-8 shows the formulation of the ginseng oral film preparation. The main ingredient of the ginseng oral film preparation is ginseng extract, with capric acid as a penetration enhancer, HPMC as the main film-forming material, carbomer as an adhesive, and propylene glycol as a plasticizer. [Table 23] 3.3.6 Optimal Manufacturing Process First, the carbomer was swollen for 12 hours, the film solution was heated to 90°C in a water bath, then hydroxypropyl methylcellulose was added and dissolved for 1 hour, and capric acid was dissolved in propylene glycol and added to the film solution. After the film solution cooled to room temperature, ginseng extract was added, stirred until dissolved, and left to stand for use. Ethyl cellulose was dissolved in anhydrous ethanol, and 0.81 g / cm³ was added. 3 The mixture was injected into a petri dish at a density and vacuum-dried at 60°C for 30 minutes to prepare a backing layer. Next, the drug-containing film solution was injected into the backing layer and vacuum-dried at 70°C for 2 hours to obtain a ginseng oral film preparation. Rg1 is thermally unstable, and decomposition occurs with increasing heating time; therefore, 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 Formulation This study conducted an initial evaluation of the quality of a ginseng-based oral film formulation. 4.1 Reagents and Equipment 4.1.1 Experimental materials and reagents Table 4-1 shows the materials and reagents used in this experiment. [Table 24] 4.1.2 Laboratory Equipment Table 4-2 shows the experimental equipment used in this experiment. [Table 25]

[0068] 4.2 Method 4.2.1 Visual Inspection The ginseng oral film was visually inspected for any obvious air bubbles, deformation, or uneven coloration. 4.2.2 Microscopic examination Using a cold-field emission scanning electron microscope (SU 8010), the microscopic structural morphology of the oral film surface was obtained by impacting the surface of the oral film under high vacuum and electron beam irradiation conditions, including whether or not there were crystals of drug precipitated on the surface and whether or not the structure was complete. The obtained data was corroborated with other properties of the oral film, such as mechanical properties and solubility. A blank oral film without drug support and a ginseng oral film were pre-treated by placing them in a vacuum drying box at 40°C for 12 hours to ensure the evaporation of moisture from the oral film. The dried oral film was then bonded to a metal base with a conductive adhesive, and gold was sprayed onto the surface of the film to obtain good conductivity. The microscopic structural morphology of the surfaces of the two types of oral film was then examined. 4.2.3 Weight Difference Limit Test Twenty tablets of ginseng oral film preparation were taken, the total weight was precisely measured, and the average weight was determined. Then, the weight of each tablet was precisely measured, and the weight of each tablet was compared to the average weight to examine the weight difference. 4.2.4 In vitro dwell time testing To simulate the time that an oral film can remain in the oral cavity after being affected by mechanical factors such as salivary flow and rinsing, the following apparatus was specially manufactured. The upper part is a linear reciprocating mechanical device with a slide glass fixed to an extendable lever. The slide glass is thoroughly coated with a 5% (w / w) mucin solution and used to simulate the surface of the oral mucosa. Cut blank oral film without drug-supported material and the drug-containing layer of ginseng oral film are attached to its surface, and a 2g weight is placed on the surface of the film for 30 seconds. The lower part uses artificial saliva as the test medium, and the slide glass moves up and down. One cycle is calculated 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 cycles / min, and the time it takes for the film to completely separate from the slide glass is recorded to simulate the residence time of the oral film in the oral cavity. 4.2.5 Tensile Strength Test In accordance with the requirements of the National Standard GBT 1040.2-2006 of the People's Republic of China, blank oral film preparations without drug support and ginseng oral film preparations were cut into dumbbell shapes using a dumbbell cutter. The cut film preparations were fixed at a gauge distance of 40 mm between two clamps of a WDT-10 electronic universal tester, and the oral film preparations were stretched longitudinally at a speed of 50 mm / min until they broke. Six samples were measured. The calculation formulas are shown in Equations 5 and 6.

number

number

[0069] 4.3 Results 4.3.1 Appearance of the film The appearance of the ginseng oral film preparation is shown in Figure 1. From Figure 1, it can be seen that the obtained ginseng oral film preparation was complete and smooth, had a uniform thickness, uniform color, no obvious air bubbles, a pale yellow surface, and no particular odor. 4.3.2 Microscopic examination The results were examined using an electron microscope. When the film was placed under a cold-field emission scanning electron microscope and observed under a 1000x magnification field, the surfaces of the blank oral film without drug support and the ginseng oral film were dense, smooth, and flat. No pores, bubbles, wrinkles, or drug crystals were observed, and there was no clear difference in the surface morphology of the two. 4.3.3 Weight Difference Limit Test Table 4-3 shows the weight difference limit requirements for the film formulation. The average weight range of the ginseng oral film formulation is 0.02g to 0.2g, and the difference limit range needs to be controlled to within 10%. The results of the weight difference limit test are shown in Table 4-4. Twenty tablets of the ginseng oral film preparation were tested and measured, and the weight difference limit for the ginseng oral film preparation was within 10%. This result indicates that the weight difference limit for the ginseng oral film preparation meets the requirements. [Table 26] [Table 27] 4.3.4 In vitro dwell time testing For Rg1 to be released from the ginseng extract, the ginseng oral film formulation needs to remain in the oral cavity for an extended period, allowing the drug to penetrate the buccal mucosa and enter the bloodstream. In this study, the in vitro residence time of the ginseng oral film formulation was significantly extended (p<0.05). Experiments showed that erosion of the film formulation by the solution began from the periphery and progressed towards the center. Furthermore, due to its own in vitro adhesive properties, the drug-carrying film formulation could remain in the oral cavity for more than 3 hours, simulating the rinsing process by saliva. The results indicate that the addition of a backing layer effectively prevents erosion of the film formulation by the solution, extends the in vitro residence time, and ensures the release and penetration of Rg1. 4.3.5 Tensile Strength Test Table 4-5 shows the stress curves and tensile strength results for the oral film formulations. The results show that the addition of ginseng extract reduced the maximum resistance of the film formulation (p<0.01) and significantly reduced the tensile strength (p<0.001). The elongation at break of the ginseng oral film formulation increased significantly (p<0.01), and the tensile strength test results indicate that the original uniform structure of the blank film formulation was destroyed after the drug was loaded. [Table 28] Compared to the blank oral film formulation, *p<0.05, **p<0.01, and ***p<0.001 were observed. 4.3.6 Toughness Test The results of the toughness tests are shown in Table 4-6. Both types of film agents showed some resistance to surface compressive force, but the puncture resistance of the drug-loaded film agent was significantly reduced (p<0.05). Both the puncture and tensile tests showed that the oral film agent destroyed its own structure after loading the drug, reflecting differences in mechanical properties. [Table 29] Note: * The p-value was < 0.05 compared to the puncture force of the blank film agent. 4.3.7 Wettability Test Table 4-7 shows the results of the wettability test. When the wetting angle of the blank oral film and the ginseng oral film is less than 90°, it indicates that the surface is hydrophilic and has good wettability, which is favorable for the release of Rg1 in the film. Here, the wetting angle of the ginseng oral film was significantly increased compared to the wetting angle of the blank oral film (p<0.05), which indicates that the addition of the extract increased the hydrophobicity of the surface, which may be related to the extraction method of the ginseng extract. Ginseng was extracted with 70% ethanol, and as a result, the extract had a certain degree of hydrophobicity and an increased wetting angle. [Table 30] Note: * The p-value was < 0.05 compared to the wetting angle of the blank film agent. 4.3.8 Measurement of Content Table 4-8 shows the Rg1 content of the ginseng oral film preparation. RSD < 5% indicates that the Rg1 content in the oral film preparation 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 formulation in the artificial saliva elution medium was over 80%, and the cumulative dissolution rate at 60 min was approximately 88.57%, indicating good dissolution. 4.3.10 Stability Influence Factor Test (1) High temperature test The results of the high-temperature test are shown in Table 4-9. Under high-temperature conditions of 60°C, the film material bent and lost weight. This is because moisture is lost from the film material under high-temperature conditions, and the Rg1 content did not differ from that of 0d. Therefore, the results of the high-temperature test indicate that the film material should 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 conditions of a temperature of 25°C and a humidity of 90% ± 5%, the film material underwent severe deformation, with shrinkage of all four sides, adhesion occurring between the oral film materials at 10d, and the film material becoming soft. There was no difference in Rg1 content compared to 0d. The results of the high humidity test indicate that the film material undergoes significant changes in appearance under high humidity conditions, making it unsuitable for storage and potentially causing deterioration of the film material in a humid environment. Therefore, the film material should be stored in a sealed container in a dry place. [Table 33] (3) Light irradiation test The results of the light irradiation test are shown in Table 4-11. Under strong light conditions of 4500 lx ± 500 lx, the shape of the film material changed, with both ends curling towards the middle, a decrease in weight, and no difference in Rg1 content compared to 0d. Therefore, the results of the light irradiation test indicate that the film material needs to be stored in a light-shielded environment. [Table 34]

[0070] 4.4 Summary The manufactured ginseng oral film formulation exhibited a weight difference limit that met the standard, uniform content, and, upon observation with a scanning electron microscope, was smooth and flat with no drug crystals. It could remain in a simulated in vitro oral environment for more than 3 hours, possessed good mechanical properties, and was easy to cut, use, and transport. The dissolution rate reached over 80% within 1 hour, and it should be stored in a cool, dry place.

[0071] Test Example 5: Pharmacokinetic experiment of ginseng oral film formulation 5.1 Materials and Equipment 5.1.1 Laboratory animals Twelve male golden hamsters, weighing 110±10g each, were purchased from Changchun Changsheng Biotechnology Co., Ltd. License number: NO.SCXK(Liao)2020-0001. They were reared for one week in the school's animal room at a temperature of (22±2)℃, with 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 substance solutions Preparation of Rg1 standard solution: 1.0 mg of Rg1 standard substance was accurately weighed, dissolved in methanol, and brought to a volume of 10 mL in a volumetric flask to prepare a stock solution of the standard substance with a concentration of 100 μg / mL. It was diluted to the desired concentration before use. Preparation of the internal standard substance, sanquinoside R1 standard substance (IS): 1.0 mg of sanquinoside R1 was accurately weighed, dissolved in methanol, and brought to a fixed volume in a 10 mL volumetric flask to prepare the stock solution of sanquinoside R1. Immediately before use, it was diluted with methanol to 5 μg / mL. 5.2.1 Chromatography and Mass Spectrometry Conditions The chromatography and mass spectrometry conditions are shown in Tables 5-3 and 5-4. [Table 37] [Table 38] 5.2.3 Processing of Plasma Samples 100 μL of plasma sample was taken, 20 μL of IS solution (R 15 μg / mL) was added, and after homogeneous mixing, 1000 μL of methanol (the extraction solvent) was added, and after homogeneous mixing by vortexing for 10 minutes, the mixture was centrifuged for 15 minutes (15000 rpm). All supernatant was taken and dried using a nitrogen blower. 200 μL of methanol was added to redissolve the solution, and after homogeneous mixing by vortexing for 5 minutes, all the solution was taken, filtered through a 0.22 μm filtration membrane, and the sample was injected for analysis. 5.2.4 Consideration of Methodology (1) Consideration of linear relationships 50 μL of blank plasma was taken, and 50 μL of mixed standard substance solutions of different concentrations were added to adjust the concentrations to 0.1, 0.5, 1, 5, 10, 50, 100, 500, and 1000 ng / mL, respectively. The mixture was then homogenized and processed according to the plasma processing method described in section 2.1.3. Linear regression was performed with Y as the ratio of Rg1 peak area to R1 peak area and X as the sample concentration. (2) Consideration of stability Blank plasma was collected, and quality control plasma samples containing Rg1 at concentrations of 5, 10, and 500 ng / mL were prepared. Short-term stability of the plasma samples was examined by leaving them at room temperature for 24 hours, autosampler stability was examined by storing them in an autosampler for 24 hours, freeze-thaw cycle stability was examined by repeating freeze-thaw cycles three times at -80°C, and long-term stability of the samples was examined by freezing them at -80°C for 7 days. (3) Consideration of precision and accuracy Blank plasma was collected, and samples at low, medium, and high concentrations were prepared. These samples were processed according to the plasma sample preparation method described in section 2.1.3 and injected into UHPLC-MS / MS for analysis. Three analytical batches were examined sequentially, and six samples were prepared for each concentration in each batch. The peak areas of Rg1 and IS were recorded, and the intraday and intermittent RSD values ​​for Rg1 and IS were calculated. (4) Consideration of the matrix effect Three concentrations of Rg1 standard solution (5, 10, and 500 ng / mL) were prepared, processed 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 areas 1 of Rg1 and IS were recorded. The supernatant of blank plasma was taken and prepared to the same three concentrations (5, 10, and 500 ng / mL), processed 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 areas 2 of Rg1 and IS were recorded. The matrix effect of Rg1 and IS was calculated using the ratio of peak area 2 to peak area 1. (5) Analysis of the recovery rate Blank plasma was taken and processed according to the plasma sample processing method in section "2.1.3" under conditions without the addition of IS. After drying by blowing with nitrogen, Rg1 and internal standard solutions of the corresponding concentrations were added to adjust the concentrations to 5, 10, and 500 ng / mL. Six samples of each type were prepared in parallel, and the peak areas 3 of Rg1 and IS were recorded. The recovery rate was calculated using the ratio of peak area 3 to peak area 2 in section "2.1.4.4". 5.2.3 In vivo pharmacokinetic studies in golden hamsters During a 7-day adaptive rearing period, the cheek pouches of the golden hamsters were cleaned daily with a cotton swab dipped in saline to maintain cleanliness. Twelve healthy male golden hamsters weighing between 100 and 120 g were randomly divided into two groups: an intragastric administration group and an oral-buccal mucosal administration group. The hamsters were fasted for 24 hours prior to the experiment, and their cheek pouches were cleaned with a cotton swab dipped in saline. Each group of golden hamsters was administered 500 mg / kg of ginseng extract and an equal amount of ginseng oral film (simultaneously placed in the left and right cheek pouches of the golden hamsters). 0.3 mL of blood was collected from the orbital venous plexus at 0, 0.08, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 5, and 6 hours after administration and placed in a tube moistened with heparin sodium saline. The collected whole blood was centrifuged (15000 rpm, 10 min), 100 μL of the supernatant was taken and processed according to section "2.1.3", measured according to the conditions in "2.1.2", and the data was recorded. 5.2.4 Data Processing Using the DAS 2.0 software, the main pharmacokinetic parameters were calculated according to the one-compartment model method and are shown in Table 5-8. The formula for calculating 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 pouches of a golden hamster are opened, a rich network of capillaries and a large area of ​​buccal mucosa are observed. The ginseng oral film formulation adheres to the buccal mucosa and cannot be expelled, making it an excellent model for the pharmacokinetics of oral film formulations. 5.3.2 Results of consideration of UHPLC-MS / MS methodology (1) Calibration curve Ion chromatograms of Rg1 and R1 in golden hamster plasma detected by UHPLC-MS / MS showed that Rg1 and R1 exhibited sharp chromatographic peak shapes under the chromatographic and mass spectrometry conditions described in section "2.1.2". Simultaneously, endogenous substances or metabolites in the plasma did not interfere with the measurement of the target components, and the retention times for Rg1 and R1 were 3.81 min and 3.65 min, respectively. The regression equation was Y = 770.44X - 144.05, indicating a good linear relationship between Rg1 in golden hamster plasma and Rg1 within the range of 0.1 to 1000 ng / mL. 2 The result was 0.9999. (2) Consideration of stability Table 5-5 shows the stability analysis and experimental results. The results indicate that the stability of plasma samples under the four processing methods met the relevant pharmacopoeia requirement of RSD < 10%, demonstrating the rationality of the plasma sample processing method and the UHPLC-MS / MS detection method. [Table 39] (3) Consideration of precision The precision analysis and experimental results are shown in Table 5-6. Both the intraday precision and intermittent precision of Rg1 were less than 3%, which is in accordance with the pharmacopoeia's requirements. [Table 40] (4) Consideration of recovery rate and matrix effect The results of the analysis of the matrix effect are shown in Table 5-7. From the table, the recovery rate of this 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 Figure 3 shows the mean plasma Rg1 concentration-time curves after administering the same dose of ginseng extract (500 mg / kg) intragastricly and orally and buccally to golden hamsters. Compared to the intragastric administration group, the area under the Rg1 concentration-time curve (AUC) was significantly increased in the orally and buccally administered group. In the case of intragastric administration, a second drug peak appeared at approximately 1.5 hours. This is because interconversion between ginsenosides occurred in the acidic environment of the golden hamster's gastrointestinal tract, increasing the Rg1 content in the plasma. Further research is needed to understand the specific reason for the appearance of the second drug peak. 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 Table 5-8. After administration of the same dose of ginseng extract, the AUC of the measured component Rg1 in plasma was measured. (0-t) The drug half-life of Rg1 in golden hamsters was significantly lower in the intragastric administration group than in the oral-buccal administration group (p<0.001). Compared to the intragastric administration group, the oral-buccal administration group significantly prolonged the drug half-life of Rg1 (p<0.05). However, C max There was no significant difference in AUC. (0-t) The results show that the oral-buccal mucosal administration group performed better than the gastric administration group. This is because the ginseng oral film formulation adhered to the cheek pouches of golden hamsters for approximately 3 hours and entered the systemic blood circulation through the buccal mucosa. The relative bioavailability F for oral-buccal mucosal administration was 369.57%, indicating that the production of the ginseng oral film formulation achieved the purpose of the formulation. [Table 42] Compared to the intragastric administration group Rg1, * p<0.05, ** p<0.01, *** p < 0.001.

[0074] 5.4 Summary After the ginseng extract was manufactured into an oral film formulation, it adhered to the surface of the buccal mucosa of golden hamsters for an extended period, significantly prolonging the elimination half-life of Rg1 in the blood. Compared to intragastric administration, the AUC (0-t) The bioavailability increased 3.70 times, and the relative bioavailability was 369.57%, indicating that the ginseng oral film formulation can achieve the purpose of the formulation and effectively improve 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 effectively extracts ginsenoside Rg1 by reflux extraction using an aqueous ethanol solution as the extraction solvent, and by limiting the parameter range to an appropriate level so that the ginsenoside Rg1 content in the obtained ginseng extract is high, thereby reducing raw material costs and being suitable for mass production.

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

[0077] Although the above embodiments have described the present invention in detail, they represent only a portion of the present invention, not all embodiments, and other embodiments can be obtained based on these embodiments without inventive step, and all of these embodiments fall within the scope of protection of the present invention.

Claims

1. A method for producing a ginseng oral film preparation, wherein the ginseng oral film preparation comprises a backing layer and a drug-containing layer that are sequentially laminated, The drug-containing layer comprises, by mass, 10 to 12 parts hydroxypropyl methylcellulose, 0.1 to 1 part carbomer, 9 to 11 parts ginseng extract, and 0.1 to 0.3 parts fatty acid. The backing layer contains 0.6 to 0.8 parts of ethylcellulose based on the mass parts of hydroxypropyl methylcellulose. The aforementioned fatty acids include one or more of caproic acid, caprylic acid, capric acid, lauric acid, stearic acid, oleic acid, and linoleic acid. A method for producing ginseng extract includes the steps of using an aqueous ethanol solution as an extraction solvent, reflux extracting ginseng, and obtaining the ginseng extract. The mass ratio of the aforementioned ginseng to the ethanol aqueous solution is 0.5 to 1:6 to 8. The volume fraction of ethanol in the aforementioned aqueous ethanol solution is 65 to 75%. The single extraction time for the reflux extraction is 0.5 to 1.5 hours, and the number of extractions is 1 to 3. The method for producing the aforementioned ginseng oral film preparation is as follows: The first step involves sequentially injecting an ethylcellulose solution into an anti-adhesion container and performing a first molding to obtain a backing layer. The steps include swelling carbomer in water, mixing it with hydroxypropyl methylcellulose to obtain a first film solution, The steps include: performing a second mixing of the fatty acid solution and the first film solution to obtain a second film solution; The third step involves mixing the ginseng extract and the second film solution to obtain a drug-containing layer film solution. The process includes the steps of sequentially injecting the drug-containing layer film liquid into the backing layer and performing a second molding to obtain the ginseng oral film preparation, The concentration of the ethylcellulose solution is 0.01 to 0.04 g / mL, the density of the ethylcellulose solution is 0.7 to 0.9 g / cm³, the solvent of the ethylcellulose solution is ethanol, and the first molding is carried out by vacuum drying. The temperature of the first mixture is 70 to 90°C. The concentration of the fatty acid solution is 0.4 to 0.5 g / mL, and the solvent of the fatty acid solution is propylene glycol. The second molding is carried out by a vacuum drying method. The temperature of the first molding process is 50 to 70°C, and the time is 20 to 40 minutes. The second molding temperature is 60-80°C, and the time is 1-3 hours. A method for producing a ginseng oral film preparation characterized by the following:

2. The density of the drug-containing layer film liquid is 1.3 to 1.4 g / cm³. ３ The manufacturing method according to claim 1, characterized in that it is the same.

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