Nasal composite spray of murraya paniculata leaves and preparation process thereof

Abstract

The present application relates to the technical field of nasal spray, in particular to a kind of nasal use of shangxiangyuan leaf composite spray and its preparation process.The preparation process includes the following steps: the system mass ratio is 1.5-2% of nano microspheres, 2.5-3% of glycerol, 0.3-0.5% of hydroxyethyl cellulose, 0.2-0.3% of phenoxyethanol and 20-25% of citrate buffer are mixed and the rest is made up with pure water, stirred until uniform, encapsulated to obtain nasal use of shangxiangyuan leaf composite spray.The present application carries out esterification reaction of glycerophosphocholine and lauric acid to synthesize glycerophosphocholine-lauric acid ester, glycerophosphocholine-lauric acid ester is the skeleton structure with glycerol, and ester bond is formed, so that glycerophosphocholine retains the structure of glycerophosphocholine while introducing the long chain of lauric acid, has better bioavailability and lower mucous membrane irritation, and provides stable structure and encapsulation efficiency for subsequent formation of liposome coating structure and loading shangxiangyuan leaf extract.

Description

Technical Field

[0001] The invention relates to the technical field of nasal sprays, and in particular to a nasal composite spray of scutellaria baicalensis leaves and a preparation process thereof. Background Art

[0002] The leaves of the plant Cyperus rotundus are the dried leaves of the plant Cyperus rotundus. They have the effects of clearing away heat and detoxifying, relieving sore throat and swelling, promoting blood circulation and relieving pain. They can be used for antibacterial, antiviral and anti-inflammatory purposes. The leaves of Cyperus rotundus are often extracted through ethanol. The ethanol extract of the leaves of Cyperus rotundus is rich in flavonoids such as urushiol and quercetin, as well as triterpenes such as ursolic acid and phenolic acid compounds. It has significant anti-inflammatory, antibacterial, antiviral and immunomodulatory effects, and can be added to nasal sprays for use.

[0003] However, the alcohol extract of the leaves of the Chinese osmanthus fragrans faces the problems of low stability and difficulty in rapid release when administered into the nasal cavity. The main components of the alcohol extract of the leaves of the Chinese osmanthus fragrans, flavonoids, are easily oxidized and degraded, affecting the shelf life of the nasal spray. Therefore, encapsulation technology is often used to encapsulate the alcohol extract of the leaves of the Chinese osmanthus fragrans. This can slow down the oxidative degradation rate of the flavonoids in the alcohol extract of the leaves of the Chinese osmanthus fragrans. However, the encapsulation shell also makes it impossible to quickly release the active ingredients of the alcohol extract of the leaves of the Chinese osmanthus fragrans after spraying. Instead, it is necessary to wait for the encapsulation shell to decompose to release the contents. During the process of nasal administration, the excessively long decomposition time of the encapsulation shell will cause the alcohol extract of the leaves of the Chinese osmanthus fragrans to be discharged before it is completely released due to metabolic activity in the nasal cavity. Therefore, the present invention provides a nasal composite spray of the leaves of the Chinese osmanthus fragrans and a preparation process thereof to solve the problems existing in the above-mentioned prior art. Summary of the Invention

[0004] In view of the shortcomings of the existing technology, the purpose of the present invention is to provide a nasal composite spray of Scutellaria baicalensis leaves and a preparation process thereof.

[0005] A preparation process of a nasal composite spray of Rhizoma Cibotii leaves comprises the following steps:

[0006] The nanospheres, which account for 1.5-2% of the system mass, glycerin, 0.3-0.5% of hydroxyethyl cellulose, 0.2-0.3% of phenoxyethanol and 20-25% of citrate buffer are mixed and the balance is supplemented with pure water, and the mixture is stirred until uniform, and then encapsulated to obtain a nasal compound spray of Scutellaria baicalensis leaves;

[0007] The preparation method of the nanospheres is as follows: 0.5-1 parts by mass of Span 80 are added to 55-60 parts by mass of liquid paraffin, and the mixture is stirred at a speed of 200-300 r / min to obtain an oil phase; 3-5 parts by mass of hyaluronic acid grafted glycerophosphocholine-laurate and 0.5-1 parts by mass of modified Scutellaria baicalensis leaf alcohol extract are mixed and added to 10-15 parts by mass of phosphate buffer, and ultrasonically dispersed for 3-5 minutes. As the aqueous phase, the aqueous phase is added dropwise to the oil phase, and high-pressure homogenization is circulated at 50-55 MPa for 3 times to obtain colostrum; 0.1-0.3 parts by mass of genipin are added, and the mixture is stirred at 40-45°C for 2-2.5 hours. After the reaction is completed, 3 times the volume of anhydrous ethanol of the system is added, and ultrasonic demulsification is carried out in an ice-water bath at 0-4°C for 30-35 minutes. The nanospheres are centrifuged, washed, dispersed with deionized water, and freeze-dried to obtain the nanospheres.

[0008] Furthermore, the preparation method of the modified ethanol extract of the leaves of Scutellaria baicalensis comprises the following steps:

[0009] The dried Scutellaria serrata leaves are crushed, and 3 times the volume of a 70-75wt% ethanol solution is added, and the mixture is extracted at 0-4°C for 48-50h, filtered, and the extraction is repeated three times. The filtrate is concentrated by rotary evaporation to obtain an extract, and the extract is adsorbed by an adsorption resin. After the impurities are washed with water, the extract is eluted with a 70-75wt% ethanol solution, desorbed, and dried to obtain an Scutellaria serrata leaf alcohol extract. The Scutellaria serrata leaf alcohol extract and acetic anhydride are mixed at a mass ratio of 1:(2-3) and completely dissolved in pyridine, and the mixture is magnetically stirred in an oil bath at 60-65°C and reacted in the dark for 4-4.5h. Subsequently, 10-12 parts by mass of 0-4°C anhydrous ethanol is added, and the mixture is placed in an ice-water bath at 0-4°C for 10-15min to terminate the reaction. Subsequently, the reactant is rotary evaporated to remove pyridine and acetic anhydride, dialyzed for purification, and freeze-dried to obtain a modified Scutellaria serrata leaf alcohol extract.

[0010] Furthermore, the preparation method of hyaluronic acid grafted glycerophosphocholine-laurate comprises the following steps:

[0011] 9-10 parts by mass of hyaluronic acid and 1-2 parts by mass of magnesium chloride solution are added together to 20-22 parts by mass of anhydrous dimethyl sulfoxide, and stirred at 60-65° C. for 3-5 minutes to obtain an activated hyaluronic acid solution. 1-2 parts by mass of glycerophosphocholine-laurate and 0.12-0.15 parts by mass of N,N'-carbonyldiimidazole are added to 10-12 parts by mass of anhydrous dimethyl sulfoxide, and stirred at 50-55° C. for 4-4.5 hours. Then, 1-3 parts by mass of the activated hyaluronic acid solution is added, and the pH value is adjusted to 8-9 with triethylamine. The mixture is reacted at 70-75° C. under nitrogen protection for 24-25 hours. The product is precipitated, centrifuged, washed and purified to remove residual dimethyl sulfoxide and N,N'-carbonyldiimidazole, and freeze-dried to obtain hyaluronic acid-grafted glycerophosphocholine-laurate.

[0012] Furthermore, the preparation method of glycerophosphocholine-laurate comprises the following steps:

[0013] 1-1.5 parts by mass of glycerophosphocholine and 1.2-3 parts by mass of lauric acid are mixed, and then 10-12 parts by mass of anhydrous ethyl acetate is added. The mixture is cooled to 0-4°C under nitrogen protection, and then 2-2.2 parts by mass of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.5-0.8 parts by mass of imidazole are added while continuously introducing nitrogen. The mixture is then stirred at 20-25°C and a speed of 100-120 r / min for 14-15 hours. The product is filtered, washed, and purified to obtain glycerophosphocholine-laurate.

[0014] Furthermore, the purification step performed during the preparation of glycerophosphocholine-laurate is to use a silica gel column for purification and elution to remove 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride.

[0015] Furthermore, the concentration of the magnesium chloride solution is 5-8wt%.

[0016] Furthermore, the pH value of the phosphate buffer is 7.4-7.5.

[0017] Furthermore, the pH value of the citrate buffer is 6-6.5.

[0018] A nasal composite spray of Sargassum serrata leaves is prepared by the above-mentioned preparation process of the nasal composite spray of Sargassum serrata leaves.

[0019] The present invention has the following advantages:

[0020] 1. The present invention conducts an esterification reaction between glycerophosphocholine and lauric acid to synthesize glycerophosphocholine-laurate. Glycerol phosphorylation choline-laurate has glycerol as its skeleton structure, and lauric acid is connected to the hydroxyl group of glycerol through an ester bond. This structure is similar to natural phospholipids and belongs to the glycerophospholipid class of compounds. While introducing the long chain of lauric acid into glycerophosphocholine, the structure of glycerophosphocholine is retained. The glycerophospholipid structure is closer to the natural components of biological membranes, and has better bioacceptability and lower mucosal irritation. At the same time, the structure and groups of glycerophosphocholine-laurate make it easier to form a liposome structure in a solution, thereby providing a stable structure and improving the encapsulation rate for the subsequent formation of a liposome-coated structure and loading the Scutellaria serrata leaf extract.

[0021] 2. The present invention forms hyaluronic acid-grafted glycerophosphocholine-laurate by a phosphoesterification reaction with hyaluronic acid, which serves as a coating shell. The hyaluronic acid-grafted glycerophosphocholine-laurate is an amphiphilic complex that provides an ideal carrier for encapsulating the Scutellaria serrata leaf extract. The stability of the encapsulation structure is maintained by chemical bonds, electrostatic interactions, and hydrophobic interactions, maintaining integrity under storage conditions. During use, controlled release is achieved through the pressure applied by the spray. The nasal spray exerts pressure on the molecular structure under the action of mechanical stress, causing a liquid crystal phase transition, thereby causing local phase separation. This causes a burst release of the Scutellaria serrata leaf extract originally encapsulated within the glycerophosphocholine-laurate and hyaluronic acid, and promotes the diffusion of the Scutellaria serrata leaf extract. This structural design allows the Scutellaria serrata leaf extract to be isolated from the external environment and reduce the degree of oxidation during storage. During the nasal spray, a burst release occurs, increasing the concentration of the Scutellaria serrata leaf extract in the drug mist after the spray is pressed and sprayed, and allowing it to penetrate into the nasal cavity faster, completing drug release.

[0022] 3. The present invention uses acetic anhydride to modify the alcohol extract of the leaves of Scutellaria baicalensis, and reacts with the groups in the acetic anhydride in the alcohol extract of the leaves of Scutellaria baicalensis, so that the modified alcohol extract of the leaves of Scutellaria baicalensis has the characteristics of an acetylated derivative, its fat solubility is improved, and it is easier to penetrate the phospholipid bilayer structure of the nasal mucosa and release it. In addition, acetic anhydride can also be locally acidified during the enzymatic hydrolysis process of intranasal drug release, and through local acidification, it produces a synergistic effect with the alcohol extract of the leaves of Scutellaria baicalensis, inhibits the growth of pathogens, and improves the antibacterial effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a scanning electron microscope image of the nanospheres prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0024] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Example 1:

[0025] A preparation process of a nasal composite spray of Rhizoma Cibotii leaves comprises the following steps:

[0026] S1: 1 part by mass of glycerophosphocholine and 1.2 parts by mass of lauric acid were mixed, and then 10 parts by mass of anhydrous ethyl acetate was added. The mixture was cooled to 0°C under nitrogen protection, and then 2 parts by mass of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.5 parts by mass of imidazole were added while continuously introducing nitrogen. The mixture was then stirred at 20°C and 100 r / min for 14 hours. The product was filtered, washed, and purified to obtain glycerophosphocholine monolaurate.

[0027] The purification was performed using a silica gel column for purification and elution to remove 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride.

[0028] S2: 9 parts by mass of hyaluronic acid and 1 part by mass of 5 wt% magnesium chloride solution were added to 20 parts by mass of anhydrous dimethyl sulfoxide, and stirred at 60°C for 3 minutes to obtain an activated hyaluronic acid solution. 1 part by mass of glycerophosphocholine-laurate and 0.12 parts by mass of N,N'-carbonyldiimidazole were added to 10 parts by mass of anhydrous dimethyl sulfoxide, stirred at 50°C for 4 hours, and then 1 part by mass of activated hyaluronic acid solution was added. The pH value was adjusted to 8 with triethylamine, and the reaction was carried out at 70°C under nitrogen protection for 24 hours. The product was precipitated, centrifuged, washed and purified to remove residual dimethyl sulfoxide and N,N'-carbonyldiimidazole, and freeze-dried to obtain hyaluronic acid grafted glycerophosphocholine-laurate.

[0029] S3: After the dried Scutellaria baicalensis leaves are crushed, 3 times the volume of 70wt% ethanol solution is added and extracted at 0℃ for 48h, filtered, and the extraction is repeated three times. The liquid part is rotary evaporated and concentrated to obtain an extract. The extract is adsorbed with an adsorption resin, and the impurities are washed with water and then eluted with 70wt% ethanol solution. Desorption and drying are performed to obtain the alcohol extract of Scutellaria baicalensis leaves. The alcohol extract of Scutellaria baicalensis leaves and acetic anhydride are mixed in a mass ratio of 1:2 and completely dissolved in pyridine. The mixture is magnetically stirred in an oil bath at 60℃ and reacted in the dark for 4h. Subsequently, 10 parts by mass of 0℃ anhydrous ethanol are added and the mixture is placed in an ice-water bath at 0℃ for 10min to terminate the reaction. The reactants are then rotary evaporated to remove pyridine and acetic anhydride, purified by dialyze, and freeze-dried to obtain the modified Scutellaria baicalensis leaf alcohol extract.

[0030] S4: Add 0.5 parts by mass of Span 80 to 55 parts by mass of liquid paraffin, stir and mix at a speed of 200r / min to obtain an oil phase, then mix 3 parts by mass of hyaluronic acid grafted glycerophosphocholine-laurate and 0.5 parts by mass of modified Scutellaria baicalensis leaf alcohol extract and add them to 10 parts by mass of phosphate buffer, the pH value of the phosphate buffer is 7.4, ultrasonic dispersion for 3 minutes, as the aqueous phase, the aqueous phase is added dropwise to the oil phase, 50MPa high-pressure homogenization cycle 3 times to obtain colostrum, then add 0.1 parts by mass of genipin, stir at 40℃ for 2h, after the reaction is completed, add 3 times the volume of anhydrous ethanol of the system, and ultrasonically demulsify in an ice-water bath at 0℃ for 30min, centrifuge, wash, add deionized water dispersion and freeze-dry to obtain nanoparticles, and use SEM to observe the surface morphology of the nanoparticles, as shown Figure 1 shown.

[0031] S5: Mix 1.5% of nanospheres, 2.5% of glycerol, 0.3% of hydroxyethyl cellulose, 0.2% of phenoxyethanol and 20% of citrate buffer solution by mass, and make up the balance with pure water, the pH value of the citrate buffer solution is 6. Then stir until uniform, and encapsulate to obtain the nasal Scutellaria baicalensis leaf composite spray. Example 2:

[0032] A preparation process of a nasal composite spray of Rhizoma Cibotii leaves comprises the following steps:

[0033] S1: 1.5 parts by mass of glycerophosphocholine and 3 parts by mass of lauric acid were mixed, and then 12 parts by mass of anhydrous ethyl acetate was added. The mixture was cooled to 0°C under nitrogen protection, and then 2.2 parts by mass of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.8 parts by mass of imidazole were added while continuously introducing nitrogen. The mixture was then stirred at 20°C and 100 r / min for 14 hours. The product was filtered, washed, and purified to obtain glycerophosphocholine monolaurate.

[0034] The purification was performed using a silica gel column for purification and elution to remove 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride.

[0035] S2: 10 parts by mass of hyaluronic acid and 2 parts by mass of 8wt% magnesium chloride solution were added to 22 parts by mass of anhydrous dimethyl sulfoxide, and stirred at 60°C for 3 minutes to obtain an activated hyaluronic acid solution. 2 parts by mass of glycerophosphocholine-laurate and 0.15 parts by mass of N,N'-carbonyldiimidazole were added to 12 parts by mass of anhydrous dimethyl sulfoxide, stirred at 50°C for 4 hours, and then 3 parts by mass of activated hyaluronic acid solution were added. The pH value was adjusted to 8 with triethylamine, and the reaction was carried out at 70°C under nitrogen protection for 24 hours. The product was precipitated, centrifuged, washed and purified to remove residual dimethyl sulfoxide and N,N'-carbonyldiimidazole, and freeze-dried to obtain hyaluronic acid grafted glycerophosphocholine-laurate.

[0036] S3: After the dried Scutellaria baicalensis leaves are crushed, 3 times the volume of 75wt% ethanol solution is added and extracted at 0℃ for 48h, filtered, and the extraction is repeated three times. The liquid part is rotary evaporated and concentrated to obtain an extract. The extract is adsorbed with an adsorption resin, and the impurities are washed with water and then eluted with 75wt% ethanol solution. Desorption and drying are performed to obtain the alcohol extract of Scutellaria baicalensis leaves. The alcohol extract of Scutellaria baicalensis leaves and acetic anhydride are mixed in a mass ratio of 1:3 and completely dissolved in pyridine. The mixture is magnetically stirred in an oil bath at 60℃ and reacted in the dark for 4h. Subsequently, 12 parts by mass of 0℃ anhydrous ethanol are added and placed in an ice-water bath at 0℃ for 10min to terminate the reaction. The reactants are then rotary evaporated to remove pyridine and acetic anhydride, purified by dialyze, and freeze-dried to obtain the modified Scutellaria baicalensis leaf alcohol extract.

[0037] S4: Add 1 part by mass of Span 80 to 60 parts by mass of liquid paraffin, stir and mix at a speed of 200 r / min to obtain an oil phase, then mix 5 parts by mass of hyaluronic acid grafted glycerophosphocholine-laurate and 1 part by mass of modified Scutellaria baicalensis leaf alcohol extract and add them to 15 parts by mass of phosphate buffer, the pH value of the phosphate buffer is 7.4, ultrasonic dispersion is carried out for 3 minutes, as the aqueous phase, the aqueous phase is added dropwise to the oil phase, 50 MPa high-pressure homogenization cycle is repeated 3 times to obtain colostrum, then add 0.3 parts by mass of genipin, stir at 40°C for 2 hours, add 3 times the volume of anhydrous ethanol of the system after the reaction is completed, and ultrasonically break the emulsion in an ice-water bath at 0°C for 30 minutes, centrifuge, wash, add deionized water for dispersion, and freeze-dry to obtain nanospheres.

[0038] S5: Mix 2% of the system mass of nano-microspheres, 3% of glycerin, 0.5% of hydroxyethyl cellulose, 0.3% of phenoxyethanol and 25% of citrate buffer solution, and make up the balance with pure water, the pH value of the citrate buffer solution is 6, then stir until uniform, and encapsulate to obtain the nasal Scutellaria baicalensis leaf composite spray. Example 3:

[0039] A preparation process of a nasal composite spray of Rhizoma Cibotii leaves comprises the following steps:

[0040] S1: 1 part by mass of glycerophosphocholine and 1.2 parts by mass of lauric acid were mixed, and then 10 parts by mass of anhydrous ethyl acetate was added. The mixture was cooled to 4°C under nitrogen protection, and then 2 parts by mass of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.5 parts by mass of imidazole were added while continuously introducing nitrogen. The mixture was then stirred at 25°C and 120 r / min for 15 hours. The product was filtered, washed, and purified to obtain glycerophosphocholine monolaurate.

[0041] The purification was performed using a silica gel column for purification and elution to remove 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride.

[0042] S2: 9 parts by mass of hyaluronic acid and 1 part by mass of 5 wt% magnesium chloride solution were added to 20 parts by mass of anhydrous dimethyl sulfoxide, and stirred at 65°C for 5 minutes to obtain an activated hyaluronic acid solution. 1 part by mass of glycerophosphocholine-laurate and 0.12 parts by mass of N,N'-carbonyldiimidazole were added to 10 parts by mass of anhydrous dimethyl sulfoxide, and stirred at 55°C for 4.5 hours. Then, 1 part by mass of activated hyaluronic acid solution was added, and the pH value was adjusted to 9 with triethylamine. The reaction was carried out at 75°C under nitrogen protection for 25 hours. The product was precipitated, centrifuged, washed and purified to remove residual dimethyl sulfoxide and N,N'-carbonyldiimidazole, and freeze-dried to obtain hyaluronic acid grafted glycerophosphocholine-laurate.

[0043] S3: After the dried Scutellaria baicalensis leaves are crushed, 3 times the volume of 70wt% ethanol solution is added and extracted at 4°C for 50h, filtered, and the extraction is repeated three times. The liquid part is rotary evaporated and concentrated to obtain an extract. The extract is adsorbed with an adsorption resin, and the impurities are washed with water and then eluted with 75wt% ethanol solution. Desorption and drying are performed to obtain the Scutellaria baicalensis leaf alcohol extract. The Scutellaria baicalensis leaf alcohol extract and acetic anhydride are mixed in a mass ratio of 1:2 and completely dissolved in pyridine. The mixture is magnetically stirred in an oil bath at 65°C and reacted in the dark for 4.5h. Subsequently, 10 parts by mass of 4°C anhydrous ethanol are added and the mixture is placed in a 4°C ice-water bath for 15min to terminate the reaction. The reactants are then rotary evaporated to remove pyridine and acetic anhydride, purified by dialyze, and freeze-dried to obtain the modified Scutellaria baicalensis leaf alcohol extract.

[0044] S4: Add 0.5 parts by mass of Span 80 to 55 parts by mass of liquid paraffin, stir and mix at a speed of 300 r / min to obtain an oil phase, then mix 3 parts by mass of hyaluronic acid grafted glycerophosphocholine-laurate and 0.5 parts by mass of modified Scutellaria baicalensis leaf alcohol extract and add them to 10 parts by mass of phosphate buffer, the pH value of the phosphate buffer is 7.5, ultrasonic dispersion is carried out for 5 minutes, as the aqueous phase, the aqueous phase is added dropwise to the oil phase, 55 MPa high-pressure homogenization cycle is repeated 3 times to obtain colostrum, then add 0.1 parts by mass of genipin, stir at 45°C for 2.5 hours, add 3 times the volume of anhydrous ethanol of the system after the reaction is completed, and ultrasonically break the emulsion in an ice-water bath at 4°C for 35 minutes, centrifuge, wash, add deionized water for dispersion, and freeze-dry to obtain nanospheres.

[0045] S5: Mix 1.5% of nanospheres, 2.5% of glycerol, 0.3% of hydroxyethyl cellulose, 0.2% of phenoxyethanol and 20% of citrate buffer solution, and make up the balance with pure water. The pH value of the citrate buffer solution is 6.5. Then stir until uniform, and encapsulate to obtain the nasal Scutellaria baicalensis leaf composite spray.

[0046] Comparative Example 1:

[0047] Compared with Example 1, the difference of Comparative Example 1 is that step S1 is not performed, glycerophosphocholine-laurate is not added in step S2, but glycerophosphocholine is used to replace glycerophosphocholine-laurate, and the other steps remain unchanged. The prepared nasal Scutellaria baicalensis leaf composite spray is recorded as Comparative Example 1.

[0048] Comparative Example 2:

[0049] Compared with Example 1, the difference of Comparative Example 2 is that step S1 is not performed, glycerophosphocholine-laurate is not added in step S2, but glycerophosphocholine-laurate is replaced by laurate, and the other steps remain unchanged. The prepared nasal Scutellaria baicalensis leaf composite spray is recorded as Comparative Example 2.

[0050] Comparative Example 3:

[0051] Compared with Example 1, the difference of Comparative Example 3 is that step S2 is not performed, and hyaluronic acid-grafted glycerophosphocholine-laurate is not added in step S4. Instead, hyaluronic acid-grafted glycerophosphocholine-laurate is replaced by glycerophosphocholine-laurate, and the other steps remain unchanged. The prepared nasal Scutellaria baicalensis leaf composite spray is recorded as Comparative Example 3.

[0052] Comparative Example 4:

[0053] Compared with Example 1, the difference of Comparative Example 4 is that acetic anhydride is not added in step S3, and the other steps remain unchanged. The prepared Scutellaria baicalensis leaf composite spray for nasal use is recorded as Comparative Example 4.

[0054] The encapsulation efficiency of the nano-microspheres in the nasal composite spray of the Rhizoma Cyperi leaves of the test examples 1-3 and comparative examples 1-2 is shown in Table 1.

[0055] Examples 1-3 and Comparative Example 3 were respectively filled in spray bottles of the same model and the same pressure, and sprayed on the surface of a glass sheet. The liquid on the surface of the glass sheet was collected, and the content of the flavonoids in the alcohol extract of the Chinese fragrant roundworm in the water liquid on the surface of the glass sheet was immediately tested. Then, Examples 1-3 and Comparative Example 3 were coated on the surface of the glass sheet in the same coating amount as the volume of the liquid on the surface of the glass sheet in the spraying experiment. After 5 seconds of coating, the content of the flavonoids in the alcohol extract of the Chinese fragrant roundworm in the water liquid on the surface of the glass sheet was tested, as shown in Table 2.

[0056] Examples 1-3 and Comparative Example 4 were subjected to an antibacterial test: Staphylococcus aureus and its bacterial solution were cultured, and Examples 1-3 and Comparative Example 4 were added to the bacterial solution. The culture was carried out at 37°C for 24 hours, the number of colonies was counted, and the antibacterial rate was calculated. The results are shown in Table 3.

[0057] Table 1

[0058]

[0059] Table 2

[0060]

[0061] Table 3

[0062]

[0063] As can be seen from Table 1, the encapsulation efficiency of Examples 1-3 is above 65.4%, while the encapsulation efficiency of Comparative Example 1 and Comparative Example 2 is only 51.3% and 52.8%, respectively. It can be seen that the nanospheres prepared by the present invention have a good encapsulation efficiency, and the raw material combination used in the present invention has a synergistic effect compared with a single raw material, further improving the encapsulation efficiency.

[0064] As can be seen from Table 2, the main component in the leaf of Melia azedarach is flavonoids, which is the signature component of the leaf of Melia azedarach, and no other flavonoids are added in the nasal spray prepared in the application, thus the flavonoid content can reflect the amount of the leaf of Melia azedarach ethanol extract, and the flavonoid content in the water solution of Examples 1-3 is higher when sprayed by spraying than by coating, which shows that the leaf of Melia azedarach ethanol extract encapsulated in the nano-microspheres is released under the pressure, and is immediately released from the nano-microspheres, while the coating method does not release, which shows that the leaf of Melia azedarach ethanol extract of Examples 1-3 does not immediately release without pressure, but slowly releases with the disintegration of the nano-microspheres, and when sprayed by the spray bottle, the release amount is increased due to the pressure response, surface change and the like, while the flavonoid content of Comparative Example 3 is 19.1% and 18.2% respectively in the spraying experiment and the coating experiment, and the change amount is small, which shows that the pressure response burst release ability is poor, and it is shown that under the condition of Comparative Example 3, since the nano-microspheres are not composed of hyaluronic acid grafted glycerophosphocholine-lauric acid ester, the ability to release the inclusions under pressure is far inferior to that of the examples.

[0065] As can be seen from Table 3, the bacteriostatic rate of Examples 1-3 is above 77.4%, while that of Comparative Example 4 is only 70.5%, which shows that the raw material combination of the application has better antibacterial effect.

[0066] It should be understood that, for those skilled in the art, improvements or changes can be made according to the above description, and all these improvements and changes shall belong to the protection scope of the appended claims of the application. The parts not described in detail in the specification belong to the prior art known to those skilled in the art.

Claims

1. A preparation process of a nasal spray of Rhizoma Cibotii leaves, characterized in that: The steps include: The nanospheres, which account for 1.5-2% of the system mass, glycerin, 0.3-0.5% of hydroxyethyl cellulose, 0.2-0.3% of phenoxyethanol and 20-25% of citrate buffer are mixed and the balance is supplemented with pure water, and the mixture is stirred until uniform, and then encapsulated to obtain a nasal compound spray of Scutellaria baicalensis leaves; The preparation method of the nanospheres is as follows: 0.5-1 parts by mass of Span 80 are added to 55-60 parts by mass of liquid paraffin, and the mixture is stirred at a speed of 200-300 r / min to obtain an oil phase, and then 3-5 parts by mass of hyaluronic acid grafted glycerophosphocholine-laurate and 0.5-1 parts by mass of modified Scutellaria baicalensis leaf ethanol extract are mixed and added to 10-15 parts by mass of phosphate buffer, and ultrasonically dispersed for 3-5 minutes. As the aqueous phase, the aqueous phase is added dropwise to the oil phase, and high-pressure homogenization is circulated at 50-55 MPa for 3 times to obtain colostrum, and then 0.1-0.3 parts by mass of genipin are added, and the mixture is stirred at 40-45°C for 2-2.5 hours. After the reaction is completed, 3 times the volume of anhydrous ethanol of the system is added, and ultrasonic demulsification is carried out in an ice-water bath at 0-4°C for 30-35 minutes. The nanospheres are centrifuged, washed, dispersed with deionized water, and freeze-dried to obtain the nanospheres. The preparation method of the hyaluronic acid-grafted glycerophosphocholine-laurate comprises: mixing 1-1.5 parts by mass of glycerophosphocholine and 1.2-3 parts by mass of lauric acid, then adding 10-12 parts by mass of anhydrous ethyl acetate, cooling to 0-4° C. under nitrogen protection, then adding 2-2.2 parts by mass of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 0.5-0.8 parts by mass of imidazole, while continuously introducing nitrogen, and then stirring at 20-25° C. and a speed of 100-120 r / min for 14-15 hours. The product is filtered, washed, and purified to obtain glycerophosphocholine-laurate; 9-10 parts by mass of hyaluronic acid and 1-2 parts by mass of magnesium chloride solution are added together to 20-22 parts by mass of anhydrous dimethyl sulfoxide, and stirred at 60-65° C. for 3-5 minutes to obtain an activated hyaluronic acid solution. 1-2 parts by mass of glycerophosphocholine-laurate and 0.12-0.15 parts by mass of N,N'-carbonyldiimidazole are added to 10-12 parts by mass of anhydrous dimethyl sulfoxide, and stirred at 50-55° C. for 4-4.5 hours. 1-3 parts by mass of the activated hyaluronic acid solution are then added, and the pH value is adjusted to 8-9 with triethylamine. The mixture is reacted at 70-75° C. under nitrogen protection for 24-25 hours. The product is precipitated, centrifuged, washed and purified to remove residual dimethyl sulfoxide and N,N'-carbonyldiimidazole, and freeze-dried to obtain hyaluronic acid grafted glycerophosphocholine-laurate. The preparation method of the modified Cyperus rotundus leaf ethanol extract comprises the following steps: after dried Cyperus rotundus leaf is crushed, 3 times the volume of 70-75wt% ethanol solution is added, and the mixture is extracted at 0-4°C for 48-50h, filtered, and the extraction is repeated three times. The filtrate is concentrated by rotary evaporation to obtain an extract, the extract is adsorbed by an adsorption resin, impurities are washed with water, and then eluted with 70-75wt% ethanol solution, desorbed, and dried to obtain the Cyperus rotundus leaf ethanol extract, the Cyperus rotundus leaf ethanol extract and acetic anhydride are mixed at a mass ratio of 1:(2-3) and completely dissolved in pyridine, and the mixture is magnetically stirred in an oil bath at 60-65°C and reacted in the dark for 4-4.5h, and then 10-12 parts by mass of 0-4°C anhydrous ethanol is added, and the mixture is placed in an ice-water bath at 0-4°C for 10-15min to terminate the reaction, and then the reactant is rotary evaporated to remove pyridine and acetic anhydride, dialyzed for purification, and freeze-dried to obtain the modified Cyperus rotundus leaf ethanol extract.

2. The preparation process of the nasal composite spray of Rhizoma Cibotii leaves according to claim 1, characterized in that: The purification step performed when preparing glycerophosphocholine-laurate is to use a silica gel column for purification and elution to remove 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride.

3. The preparation process of the nasal composite spray of Rhizoma Cibotii leaves according to claim 1, characterized in that: The concentration of the magnesium chloride solution is 5-8wt%.

4. The preparation process of the nasal composite spray of Rhizoma Cibotii leaves according to claim 1, characterized in that: The pH of phosphate buffer is 7.4-7.

5.

5. The preparation process of the nasal composite spray of Rhizoma Cibotii leaves according to claim 1, characterized in that: The pH of citrate buffer is 6-6.

5.

6. A nasal spray of Rhizoma Cibotii leaves, characterized in that: The nasal spray is prepared by the preparation process of the nasal safflower leaf composite spray according to any one of claims 1 to 5.

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