Perillaseed oil nanoemulsion eye drops as well as preparation method and application thereof

Abstract

The invention relates to perillaseed oil nanoemulsion eye drops as well as a preparation method and application thereof, and belongs to the technical field of medicines. The perillaseed oil nanoemulsion eye drops comprise the following raw material components in parts by mass: 1-20 parts of perillaseed oil, 0.1-10 parts of an emulsifier, 0-10 parts of a co-emulsifier, 0.05-3 parts of a metal ion chelating agent and 1-5 parts of an osmotic pressure regulator. By adding the metal ion chelating agent and controlling the preheating temperature, the problem that perillaseed oil is easy to oxidize is solved. According to the perilla oil nanoemulsion eye drops, the perilla oil serves as an effective component to exert the drug effect and also serves as an oil phase, meanwhile, the particle size of the nanoemulsion is accurately controlled by adjusting the variety of the co-emulsifier, and the safety and stability of the perilla oil nanoemulsion eye drops are further improved. The perillaseed oil eye drops provided by the invention can effectively relieve the symptoms of xerophthalmia and asthenopia, and a new medication choice is provided for treating xerophthalmia and myopia.

Description

Technical Field

[0001] The invention relates to perilla oil nanoemulsion eye drops and a preparation method and application thereof, belonging to the technical field of medicine. Background Art

[0002] Dry eye is a chronic, multifactorial ocular surface disease caused by abnormalities in tear quality, quantity, and dynamics, leading to tear film instability or an imbalance in the ocular surface microenvironment. This can be accompanied by ocular surface inflammation, tissue damage, and neurological abnormalities, resulting in a variety of ocular surface symptoms and / or visual impairment. Dyslipidemia-type dry eye is caused by abnormalities in the lipid layer, either quantitative or qualitative, such as meibomian gland dysfunction, blepharitis, and various factors that increase tear evaporation. Inflammation is also a key mechanism in the pathogenesis of dry eye. Proinflammatory cytokines such as IL-1β, IL-6, IL-17, IFN-γ, and TNF-α have been shown to accelerate the progression of dry eye. Inflammation can also directly lead to changes in the ocular surface, such as a decrease in conjunctival goblet cells, decreased mucin production, corneal epithelial abnormalities, and immune dysfunction. Therefore, anti-inflammatory treatments can effectively reverse inflammation-induced ocular surface damage, slow disease progression, and thus improve ocular surface function.

[0003] Myopia refers to a pathological condition in which parallel light rays passing through the refractive system of the eye are focused in front of the retina when the eye is at rest, resulting in decreased distance vision. The causes of myopia are complex and not yet clear, but studies have shown that it is mainly related to environmental factors (long periods of close work, reduced frequency of outdoor activities, light intensity, etc.) and genetic factors (myopia of parents). Currently, the methods commonly used to prevent and treat myopia in adolescents mainly include refractive surgery, optical correction (orthokeratology lenses, or OK lenses, frame glasses) and drug therapy. However, surgical treatment is expensive and has a variety of postoperative complications, and orthokeratology lenses are complicated to care for and prone to keratitis. Therefore, in recent years, low-cost and easy-to-use drug therapy has become a major research hotspot.

[0004] Currently, the medications used in clinical practice in my country to treat dry eye have varying degrees of side effects. Artificial tears can only alleviate dry eye symptoms, but their efficacy is limited. Among the medications used to prevent and treat myopia, only 0.01% atropine eye drops exist. However, atropine eye drops have drawbacks such as poor stability, easy hydrolysis, and the hydrolysis product, tropic acid, is mutagenic, making long-term use in adolescents unsuitable. Therefore, developing safer and more effective medications for the treatment of dry eye and myopia is of great clinical significance.

[0005] Perilla oil, extracted from the fruit of the plant Perilla frutescens, possesses a variety of pharmacological effects. Rich in unsaturated fatty acids (over 90%), perilla oil possesses strong antioxidant and anti-inflammatory properties. The α-linolenic acid content in perilla oil can range from 56.14% to 64.82%. α-linolenic acid is the metabolic starting point for ω-3 fatty acids, which are then synthesized into DHA and EPA, which have the potential to reduce inflammation and protect vital organ function. DHA, a crucial component of the retina, provides nutrients for the optic nerve and has therapeutic benefits for conditions such as fatigue and blurred vision caused by excessive eye use. However, due to its high content of unsaturated fatty acids, perilla oil is susceptible to oxidation under the influence of light, air, and temperature, leading to deterioration and difficulty in storage, which severely limits its widespread application. While nanoemulsions, as a novel drug delivery system, can improve drug bioavailability and stability and reduce drug degradation and inactivation, there are still challenges that require further research and resolution. First, while nanoemulsions can provide some protection for drugs, their oxidative stability during long-term storage and use still requires further attention. Second, the preparation of nanoemulsions typically requires complex processes and equipment. To ensure the stability and bioavailability of nanoemulsions, extensive formulation optimization is required, including the selection of appropriate emulsifiers, co-emulsifiers, and oil phases, which increases production costs and process complexity. Therefore, the development of safer and more effective perilla oil nanoemulsion products is of great practical significance. Summary of the Invention

[0006] In order to solve the above problems, the present invention provides a perilla oil nanoemulsion eye drop and its preparation method and application. The present invention uses a new plant oil (perilla oil) as the oil phase, and a mixed aqueous solution of an emulsifier, an emulsifier, a metal ion chelating agent and an osmotic pressure regulator as the aqueous phase, and prepares a new perilla oil nanoemulsion eye drop by high-speed shear-high-pressure homogenization technology. The present invention solves the problem of perilla oil being easily oxidized by adding a metal ion chelating agent and controlling the preheating temperature. The perilla oil described in the present invention is used as an effective ingredient to exert its medicinal effect and also as an oil phase. During the preparation process, in order to achieve efficient use of the medicinal effect of perilla oil, the present invention reduces the use of emulsifiers as much as possible, so that the oil phase accounts for a larger proportion. At the same time, by adjusting the type of the emulsifier, the particle size of the nanoemulsion is precisely controlled, thereby further improving the safety and stability of the perilla oil nanoemulsion eye drops.

[0007] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0008] A perilla oil nanoemulsion eye drop, comprising the following components in parts by mass:

[0009]

[0010] Preferably, the raw materials of the perilla oil nanoemulsion eye drops include the following components by mass: 5-10 parts of perilla oil, 1-1.5 parts of emulsifier, 0-1.5 parts of co-emulsifier, 0.05-1 part of metal ion chelating agent, and 2-2.5 parts of osmotic pressure regulator.

[0011] In the above technical solution, the perilla oil nanoemulsion eye drops are white emulsions with a pH of 6 to 8 and a particle size of 50 to 200 nm.

[0012] In the above technical solution, the perilla oil nanoemulsion eye drops are nanoemulsions prepared from the above raw material components, water and a pH regulator, wherein the mass ratio of the perilla oil to the nanoemulsion is 1 to 20:100.

[0013] Furthermore, the pH regulator is 0.1 mol / L sodium hydroxide.

[0014] In the above technical solution, the emulsifier is one or more of soybean lecithin S100, egg yolk lecithin E80, egg yolk lecithin PC-98T, egg yolk lecithin PL-100M, hydrogenated soybean lecithin or distearoyl phosphatidylcholine.

[0015] Preferably, the emulsifier is egg yolk lecithin E80.

[0016] In the above technical solution, the co-emulsifier is one or more of poloxamer 188, polysorbate 80, polyethylene glycol (15)-hydroxystearate, polyoxyethylene castor oil or propylene glycol.

[0017] Preferably, the co-emulsifier is polyoxyethylene castor oil.

[0018] The present invention can control the particle size of nanoparticles in the obtained perilla oil nanoemulsion eye drops by adjusting the type of the co-emulsifier. When the co-emulsifier is polyoxyethylene castor oil, the particle size of the obtained nanoparticles is smaller and more uniformly distributed, and the obtained eye drops have better stability and bioavailability.

[0019] In the above technical solution, the osmotic pressure regulator is one or more of glycerol, mannitol or sorbitol.

[0020] Preferably, the osmotic pressure regulator is glycerol.

[0021] In the above technical solution, the metal ion chelating agent is one or two of ethylenediaminetetraacetic acid, ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid or penta(carboxymethyl)diethylenetriamine; or the metal ion chelating agent is one or more of the salt or hydrate corresponding to ethylenediaminetetraacetic acid, the salt or hydrate corresponding to ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid, or the salt or hydrate corresponding to penta(carboxymethyl)diethylenetriamine.

[0022] Preferably, the metal ion chelating agent is disodium ethylenediaminetetraacetate (EDTA-2Na).

[0023] In the above technical solution, the perilla oil is taken from the natural plant Perilla frutescens, and has the advantages of being green, safe, and having no side effects.

[0024] Another object of the present invention is to provide a method for preparing the perilla oil nanoemulsion eye drops, comprising the following steps:

[0025] S1: adding an emulsifier, a co-emulsifier, a metal ion chelating agent, and an osmotic pressure regulator to purified water, stirring and dissolving them in a high-speed shearing machine at a speed of 8000-12000 rpm, and then preheating them in a magnetic stirrer at a speed of 70-90 rpm at 50°C-90°C for 5 minutes to obtain an aqueous phase;

[0026] S2: Preheat the perilla oil in a magnetic stirrer at 50°C to 90°C and 70-90 rpm for 5 minutes to obtain an oil phase. Slowly add the oil phase obtained above to the aqueous phase obtained in S1 under shearing conditions, and shear at 12,000 rpm for 5 minutes to obtain colostrum.

[0027] S3: After the colostrum cools to room temperature, dilute to 100 mL with purified water, transfer to a high-pressure homogenizer, and homogenize 8 times at 800-1000 bar at 40°C to obtain the final milk. Then, use a pH adjuster to adjust the pH to 6-8 and sterilize.

[0028] Preferably, in S1 and S2, the preheating temperature is 60°C.

[0029] In the above technical solution, in S3, high-pressure steam sterilization or sterilization through a 0.22 μm microporous filter membrane can be used.

[0030] Furthermore, the high-pressure steam sterilization condition is sterilization at 121° C. for 10 minutes.

[0031] Furthermore, when the amount of the co-emulsifier added is 0, the particle size of the nanoparticles in the obtained perilla oil nanoemulsion eye drops is relatively large, and it is preferably sterilized by high pressure sterilization.

[0032] In the above technical solution, the stirring and shearing operations are performed in existing technical devices, preferably a magnetic stirrer or a high-speed shearing machine.

[0033] Another object of the present invention is to provide the use of the perilla oil nanoemulsion eye drops in the preparation of a drug for treating dry eye.

[0034] Another object of the present invention is to provide the use of the perilla oil nanoemulsion eye drops in the preparation of a drug for treating myopia.

[0035] This invention utilizes perilla oil, a novel plant oil, not only as an oil phase but also as a pharmaceutical agent, to develop novel perilla oil nanoemulsion eye drops. The high-purity perilla oil and lipid components, such as phospholipids, used in the perilla oil nanoemulsion eye drops of the present invention can form a lipid layer on the surface of the cornea, stabilizing the tear film and preventing the volatilization of aqueous tears. This natural, non-irritating eye drop not only replenishes oil, improves dry eye, and alleviates visual fatigue, but also assists in regulating immune and inflammatory responses, while avoiding allergic reactions caused by the animal protein contained in fish oil emulsions. The successful preparation of the perilla oil eye drops in this invention provides a new medication option for treating dry eye and alleviating visual fatigue.

[0036] Beneficial effects of the present invention:

[0037] 1. The raw material perilla oil used in the present invention is taken from the natural plant Perilla frutescens, which has the advantages of being green and safe. Perilla oil is rich in unsaturated fatty acids up to more than 90%, and has strong antioxidant and anti-inflammatory activities.

[0038] 2. The raw material perilla oil used in the present invention is the vegetable oil with the highest α-linolenic acid content discovered so far, which can promote the synthesis of DHA and EPA in the body. Among them, DHA is an important component of the retina and can provide nutrients required for the visual nerves. EPA has a similar structure to arachidonic acid and can competitively inhibit the production of pro-inflammatory factors. At the same time, the α-linolenic acid in perilla oil can not only promote the development of brain nerve cells and improve memory, but also control platelet aggregation in the human body and prevent thrombosis.

[0039] 3. The perilla oil nanoemulsion eye drops obtained by the present invention use perilla oil as the oil phase and the effective ingredient to exert the medicinal effect, without adding an additional oil phase and without containing preservatives, so it is safer. Moreover, the perilla oil and lipid components can relieve dry eye symptoms and visual fatigue, and are more suitable for long-term use by teenagers.

[0040] 4. The perilla oil nanoemulsion eye drops obtained by the present invention have a simple preparation process, low technical risk, low production cost, and are easy to industrialize. In addition, the oil phase accounts for a large proportion in the preparation process, has higher stability, is easy to store and transport, and has a very broad market application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 These are the corneal sodium fluorescein staining and scoring images of mice in each group during dry eye treatment.

[0042] Figure 2 This is the relationship between the tear film breakup time (s) and time (Day) and the relationship between the tear secretion volume (mm) and time (Day) in each group of mice during dry eye treatment.

[0043] Figure 3 This figure shows the content of inflammatory factors (pg / ml) in the tears of each group of mice during dry eye treatment.

[0044] Figure 4 These are the results of the tear fern experiment on each group of mice in the treatment of dry eye.

[0045] Figure 5 These are the corneal H&E staining, conjunctival PAS staining and corneal TUNEL staining images of mice in each group during dry eye treatment.

[0046] Figure 6 These are the refractive power (D) diagrams of the experimental eyes (OS) and model eyes (OD) in each group during myopia treatment on the 0th day, the 14th day and after treatment.

[0047] Figure 7 These are the graphs of the axial length (mm) of the experimental eyes (OD) and model eyes (O) in each group during myopia treatment on the 0th day, 14th day and after treatment.

[0048] Figure 8 These are the vitreous cavity depths (mm) of the experimental eyes (OD) and model eyes (OD) in each group during myopia treatment at day 0, day 14, and after treatment.

[0049] Figure 9 These are the graphs of lens thickness and anterior chamber depth (mm) of each group of experimental eyes (OD) in myopia treatment on the 0th day, 14th day and after treatment.

[0050] Figure 10 This is a graph of the tear film breakup time (s) of each group of experimental eyes (OD) in myopia treatment on the 0th day, 14th day and after treatment. DETAILED DESCRIPTION

[0051] The following non-limiting examples may enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way.

[0052] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are all commercially available unless otherwise specified.

[0053] Example 1

[0054] Prescription: 5g perilla oil; 1.2g soybean lecithin S100; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0055] Preparation method: 1.2g soybean lecithin S100, 0.05g EDTA-2Na, and 2.2g glycerol are added to about 70mL pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 5g perilla oil is taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the high-speed shearing machine speed is set to 12000rpm, and under the shearing state, the oil phase is slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum is cooled to room temperature, pure water is added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide is used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops are obtained after sterilization through a 0.22μm filter membrane.

[0056] Example 2

[0057] Prescription: 5g perilla oil; 1.2g egg yolk lecithin E80; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0058] Preparation method: 1.2g egg yolk lecithin E80, 0.05g EDTA-2Na, and 2.2g glycerol are added to about 70mL pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 5g perilla oil is taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the high-speed shearing machine speed is set to 12000rpm, and under the shearing state, the oil phase is slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum is cooled to room temperature, pure water is added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide is used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops are obtained after sterilization through a 0.22μm filter membrane.

[0059] Example 3

[0060] Prescription: 5g perilla oil; 1.2g egg yolk lecithin PL-100M; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0061] Preparation method: 1.2g egg yolk lecithin PL-100M, 0.05g EDTA-2Na, and 2.2g glycerol were added to about 70mL of purified water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 5g perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the high-speed shearing machine speed was set to 12000rpm, and under shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum cooled to room temperature, purified water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the nanoemulsion was obtained after sterilization through a 0.22μm filter membrane.

[0062] Example 4

[0063] Prescription: 5g perilla oil; 1.2g egg yolk lecithin PC-98T; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0064] Preparation method: 1.2g egg yolk lecithin PC-98T, 0.05g EDTA-2Na, and 2.2g glycerol were added to about 70mL of purified water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 5g perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the high-speed shearing machine speed was set to 12000rpm, and under shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum cooled to room temperature, purified water was added to the volume to 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the nanoemulsion was obtained after sterilization through a 0.22μm filter membrane.

[0065] Example 5

[0066] Prescription: 5g perilla oil; 1.2g egg yolk lecithin E80; 0.8g poloxamer; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0067] Preparation method: Take 1.2g egg yolk lecithin E80, 0.8g poloxamer 188, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 5g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum cooled to room temperature, water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0068] Example 6

[0069] Prescription: 5g perilla oil; 1.2g egg yolk lecithin E80; 0.8g polysorbate-80; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0070] Preparation method: Take 1.2g egg yolk lecithin E80, 0.8g poloxamer 188, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 5g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0071] Example 7

[0072] Prescription: 5g perilla oil; 1.2g egg yolk lecithin E80; 0.8g polyoxyethylene castor oil; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0073] Preparation method: Take 1.2g egg yolk lecithin E80, 0.8g polyoxyethylene castor oil, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 5g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0074] Example 8

[0075] Prescription: 7g perilla oil; 1.2g egg yolk lecithin E80; 0.6g polyoxyethylene castor oil; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0076] Preparation method: Take 1.2g egg yolk lecithin E80, 0.6g polyoxyethylene castor oil, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 7g of perilla oil phase was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0077] Example 9

[0078] Prescription: 7g perilla oil; 1.2g egg yolk lecithin E80; 0.8g polyoxyethylene castor oil; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0079] Preparation method: Take 1.2g egg yolk lecithin E80, 0.8g polyoxyethylene castor oil, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; 7g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the water phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0080] Example 10

[0081] Prescription: 7g perilla oil; 1.2g egg yolk lecithin E80; 1.0g polyoxyethylene castor oil; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0082] Preparation method: Take 1.2g egg yolk lecithin E80, 1.0g polyoxyethylene castor oil, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 7g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0083] Example 11

[0084] Prescription: 7g perilla oil; 1.2g egg yolk lecithin E80; 1.2g polyoxyethylene castor oil; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0085] Preparation method: Take 1.2g egg yolk lecithin E80, 1.2g polyoxyethylene castor oil, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min at a speed of 70rpm in a magnetic stirrer as the aqueous phase to obtain an aqueous phase; 7g of perilla oil was taken and preheated at 60°C for 5min at a speed of 70rpm in a magnetic stirrer to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0086] Example 12

[0087] Prescription: 10g perilla oil; 1.2g egg yolk lecithin E80; 1.0g polyoxyethylene castor oil; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; purified water to 100mL.

[0088] Preparation method: Take 1.2g egg yolk lecithin E80, 1.0g polyoxyethylene castor oil, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 10g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0089] Example 13

[0090] Prescription: 10g perilla oil; 1.2g egg yolk lecithin E80; 1.2g polyoxyethylene castor oil; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0091] Preparation method: Take 1.2g egg yolk lecithin E80, 1.2g polyoxyethylene castor oil, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 10g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0092] Example 14

[0093] Prescription: 10g perilla oil; 1.2g egg yolk lecithin E80; 1.4g polyoxyethylene castor oil; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0094] Preparation method: Take 1.2g egg yolk lecithin E80, 1.2g polyoxyethylene castor oil, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 10g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 800bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.00, and the perilla oil nanoemulsion eye drops were obtained after sterilization through a 0.22μm filter membrane.

[0095] Example 15

[0096] Prescription: 10g perilla oil; 1.2g egg yolk lecithin E80; 0.05g EDTA-2Na; 2.2g glycerol; appropriate amount of sodium hydroxide; 100mL purified water.

[0097] Preparation method: Take 1.2g egg yolk lecithin E80, 0.05g EDTA-2Na and 2.2g glycerol were added to about 70mL of pure water, stirred and dissolved in a high-speed shearing machine at a speed of 12000rpm, and then preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an aqueous phase; 10g of perilla oil was taken and preheated at 60°C for 5min in a magnetic stirrer at a speed of 70rpm to obtain an oil phase; the speed of the high-speed shearing machine was set to 12000rpm, and under the shearing state, the oil phase was slowly added to the aqueous phase and sheared for 5min to obtain colostrum; after the colostrum was cooled to room temperature, pure water was added to make the volume 100mL, transferred to a high-pressure homogenizer, and homogenized 8 times at a homogenization pressure of 1000bar to obtain the final milk; 0.1mol / L sodium hydroxide was used to adjust the pH value of the final milk to 7.50, bottled, filled with nitrogen, sealed with a roll cap, sterilized with high-pressure steam at 121°C for 10min, and then quickly cooled to room temperature in a water bath after being taken out to obtain perilla oil nanoemulsion eye drops.

[0098] Characterization test of the perilla oil nanoemulsion eye drops obtained in Examples 1 to 15:

[0099] Table 1 shows the characterization results of the perilla oil nanoemulsion eye drops obtained in Examples 1 to 15. It can be seen that according to Examples 1 to 4, the particle size of the preparations obtained by using egg yolk lecithin E80 is smaller and the distribution is more uniform; according to Examples 5 to 7, polyoxyethylene castor oil can significantly reduce the particle size of the perilla oil nanoemulsion; according to Examples 8 to 11, 0.8 g of polyoxyethylene castor oil can significantly reduce the particle size of the nanoemulsion when the oil amount is 7 g; according to Examples 12 to 14, 1.0 g of polyoxyethylene castor oil can significantly reduce the particle size of the nanoemulsion when the oil amount is 10 g.

[0100] Table 1 Characterization results of perilla oil nanoemulsion eye drops obtained by different formulation processes

[0101]

[0102] Note: PDI refers to the polydispersity index, which indicates the uniformity of particle size distribution and is generally required to be less than 0.3.

[0103] Stability test of the perilla oil nanoemulsion eye drops obtained in Examples 9, 13, and 15:

[0104] (1) High temperature test: The perilla oil nanoemulsion eye drops obtained in Examples 9, 13, and 15 were placed in a constant temperature heating box at 60°C for 10 days. Samples were taken on the 5th and 10th days, respectively, and the appearance was observed. The physical and chemical properties of the preparations at each sampling time were measured and compared with those of the preparations obtained at the time of preparation. The results are shown in Table 2.

[0105] (2) Illumination test: The perilla oil nanoemulsion eye drops obtained in Examples 9, 13, and 15 were placed in a 4500±500 lx illumination box for 10 days. Samples were taken on the 5th and 10th days, respectively, and the appearance was observed. The physical and chemical properties of the preparations at each sampling time were measured and compared with those of the preparations obtained immediately after preparation. The results are shown in Table 3.

[0106] As shown in Tables 2 and 3, the perilla oil nanoemulsion eye drops prepared in Examples 9, 13, and 15 are unstable under high temperature or light conditions, and therefore should be avoided from storage under high temperature or strong light conditions.

[0107] Table 2 High temperature test results of perilla oil nanoemulsion eye drops prepared in Examples 9, 13, and 15

[0108]

[0109] Table 3 Results of illumination test of perilla oil nanoemulsion eye drops prepared in Examples 9, 13, and 15

[0110]

[0111] (3) Accelerated Test: The perilla oil nanoemulsion eye drops prepared in Examples 9, 13, and 15 were placed at 40°C ± 2°C for accelerated testing. Physical and chemical properties were measured at 0, 1, 2, 3, and 6 months. The results are shown in Table 4.

[0112] (4) Long-term test: The perilla oil nanoemulsion eye drops prepared in Examples 9, 13, and 15 were stored in the dark at room temperature below 25°C. The physical and chemical properties were evaluated at 0, 1, 3, 6, 9, and 12 months. The results are shown in Table 5.

[0113] As shown in Tables 4 and 5, the perilla oil nanoemulsion eye drops prepared in Examples 9, 13, and 15 have good stability during both accelerated storage and long-term storage.

[0114] Table 4 Accelerated test results of perilla oil nanoemulsions prepared in Examples 9, 13, and 15

[0115]

[0116] Table 5 Long-term test results of perilla oil nanoemulsion prepared in Examples 9, 13, and 15

[0117]

[0118]

[0119] Research on the perilla oil nanoemulsion eye drops obtained by the present invention in the treatment of dry eyes:

[0120] (1) Experimental groups: The mice were randomly divided into a normal group, a dry eye model group, a Soothe XP group, a 7% PFE group (the perilla oil nanoemulsion eye drops obtained in Example 9), a 10% PFE group (the perilla oil nanoemulsion eye drops obtained in Example 13), and a 10% PFE(s) group (the perilla oil nanoemulsion eye drops obtained in Example 15), with 3 mice in each group.

[0121] (2) Construction of dry eye mouse model: The mice were first adaptively fed for 3 days, and then 0.2% benzalkonium chloride solution was dripped into each eye twice a day for 1 week.

[0122] (3) Dosage regimen: Eighteen dry eye model rats were randomly divided into five groups, with three rats and six eyes in each group. Soothe XP, 7% PFE, 10% PFE, 10% PFE(s), and normal saline were slowly dripped into the lower conjunctival sac of the model mice twice a day, morning and evening, using a pipette.

[0123] (4) Experimental results: The experimental results are as follows Figures 1 to 5 As shown. Figures 1 to 5 It can be seen that perilla oil nanoemulsion eye drops and Soothe XP eye drops have a certain therapeutic effect on mice with dry eye. Each formulation group significantly reduced corneal damage, increased tear film breakup time and tear secretion volume, and restored them to normal levels. Each formulation group significantly reduced the content of inflammatory factors in tears. Each formulation group increased the production of fern crystals in tears. According to the slices, after treatment, the morphology and structure of corneal cells in each treatment group were good, with almost no cell apoptosis caused by dry eye. After treatment, the number of goblet cells in the conjunctiva of each treatment group increased significantly. In summary, this formulation shows excellent efficacy in the treatment of dry eye, and there is no significant difference compared with the marketed drug.

[0124] Research on the use of the perilla oil nanoemulsion eye drops obtained by the present invention in treating myopia:

[0125] (1) Experimental groups: The mice were randomly divided into a normal group, a myopia model group, an atropine sulfate eye drops group, a 5% PFE group (the perilla oil nanoemulsion eye drops obtained in Example 7), and a 10% PFE(s) group (the perilla oil nanoemulsion eye drops obtained in Example 15), with 5 mice in each group.

[0126] (2) Construction of a form-deprivation guinea pig myopia model: A translucent, non-toxic latex balloon was used as an eye mask to cover the left eye (OS) of the experimental group of guinea pigs, and the right eye (OD) served as the control eye. After the experiment began, the normal group did not receive any treatment, and the other four groups of guinea pigs all had their left eyes covered with latex balloons. The latex balloon was cut into a shape that fits the guinea pig's head and fixed with medical tape to ensure that the left eye is covered and the right eye is exposed. The guinea pig's left eye was checked at irregular intervals every day to confirm whether the eye mask was displaced, fell off, or blocked the contralateral eye, and whether it could completely cover the experimental eye without causing pressure on the eyeball or affecting blinking.

[0127] (3) Dosage regimen: 14 days after modeling, atropine sulfate eye drops, 5% PFE and 10% PFE(s) were dripped into the conjunctival sac of guinea pigs. Atropine sulfate eye drops were dripped once a day, and 5% PFE and 10% PFE(s) were dripped three times a day for 42 consecutive days. The modeling state was restored after each dose, for a total of 56 days.

[0128] (4) Experimental results: The experimental results are as follows Figures 6-10 As shown. Figures 6-10 It can be seen that the perilla oil nanoemulsion eye drops and atropine sulfate eye drops obtained by the present invention have a certain therapeutic effect on form deprivation guinea pig myopia, can slow down the progression of myopia in guinea pigs, and can also delay the growth of the eye axis. However, compared with atropine sulfate eye drops, the myopia effect of the perilla oil nanoemulsion eye drops provided by the present invention is relatively poor, which may be because the ω-3 unsaturated fatty acids rich in perilla oil have a low conversion rate in the body. In addition, the perilla oil nanoemulsion eye drops obtained by the present invention can increase the tear film breakup time and avoid the dry eye symptoms caused by long-term use of atropine sulfate eye drops.

Claims

1. A perilla oil nanoemulsion eye drop, characterized in that: The raw materials of the perilla oil nanoemulsion eye drops include the following components in parts by mass:

2. The perilla oil nanoemulsion eye drops according to claim 1, characterized in that: The raw materials of the perilla oil nanoemulsion eye drops include the following components by mass: 5-10 parts of perilla oil, 1-1.5 parts of emulsifier, 0-1.5 parts of co-emulsifier, 0.05-1 part of metal ion chelating agent, and 2-2.5 parts of osmotic pressure regulator.

3. The perilla oil nanoemulsion eye drops according to claim 1, characterized in that: The perilla oil nanoemulsion eye drops are white emulsions with a pH value of 6 to 8 and a particle size of 50 to 200 nm.

4. The perilla oil nanoemulsion eye drops according to claim 1, characterized in that: The perilla oil nanoemulsion eye drops are a nanoemulsion prepared from the raw material components according to claim 1, water and a pH regulator, wherein the mass ratio of the perilla oil to the nanoemulsion is 1 to 20:

100.

5. The perilla oil nanoemulsion eye drops according to claim 1 or 4, characterized in that: The pH regulator is 0.1 mol / L sodium hydroxide.

6. The perilla oil nanoemulsion eye drops according to claim 1, characterized in that: The emulsifier is one or more of soybean lecithin S100, egg yolk lecithin E80, egg yolk lecithin PC-98T, egg yolk lecithin PL-100M, hydrogenated soybean lecithin or distearoyl phosphatidylcholine; The co-emulsifier is one or more of poloxamer 188, polysorbate 80, polyethylene glycol (15)-hydroxystearate, polyoxyethylene castor oil or propylene glycol; The osmotic pressure regulator is one or more of glycerol, mannitol or sorbitol; The metal ion chelating agent is one or two of ethylenediaminetetraacetic acid, ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid or penta(carboxymethyl)diethylenetriamine; or the metal ion chelating agent is one or more of the salt or hydrate corresponding to ethylenediaminetetraacetic acid, the salt or hydrate corresponding to ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid, or the salt or hydrate corresponding to penta(carboxymethyl)diethylenetriamine.

7. The method for preparing the perilla oil nanoemulsion eye drops according to any one of claims 1 to 6, characterized in that: The steps include: S1: adding an emulsifier, a co-emulsifier, a metal ion chelating agent, and an osmotic pressure regulator to purified water, stirring and dissolving them in a high-speed shearing machine at a speed of 8000-12000 rpm, and then preheating them in a magnetic stirrer at a speed of 70-90 rpm at 50°C-90°C for 5 minutes to obtain an aqueous phase; S2: Preheat the perilla oil in a magnetic stirrer at 50°C to 90°C and 70-90 rpm for 5 minutes to obtain an oil phase. Slowly add the oil phase obtained above to the aqueous phase obtained in S1 under shearing conditions, and shear at 12,000 rpm for 5 minutes to obtain colostrum. S3: After the colostrum cools to room temperature, dilute to 100 mL with purified water, transfer to a high-pressure homogenizer, and homogenize 8 times at 800-1000 bar at 40°C to obtain the final milk. Then, use a pH adjuster to adjust the pH to 6-8 and sterilize.

8. The preparation method according to claim 7, characterized in that: In S3, high-pressure steam sterilization or sterilization through a 0.22 μm microporous filter membrane can be used, wherein the high-pressure steam sterilization condition is sterilization at 121° C. for 10 minutes.

9. Use of the perilla oil nanoemulsion eye drops according to claim 1 in preparing a medicament for treating dry eye.

10. Use of the perilla oil nanoemulsion eye drops according to claim 1 in preparing a drug for treating myopia.

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