Preparation method of cannabidiol nano-lipid carrier with transdermal absorption function

Abstract

The invention relates to a preparation method of a cannabidiol nano-lipid carrier, in particular to a preparation method of a cannabidiol nano-lipid carrier with a transdermal absorption function. The invention aims to solve the problem that the existing cannabidiol lipid carrier cannot have high water solubility and high transdermal absorption function at the same time, and is poor in stability and easy to irritate skin. The method comprises the following steps: 1, weighing raw materials; 2, preparing an oil phase; 3, preparing a water phase; 4, mixing; 5, performing ultrasonic homogenization; 6, cooling; and 7, removing impurities. The preparation method is used for preparing the cannabidiol nano-lipid carrier with the transdermal absorption function.

Description

Technical Field

[0001] The invention relates to a method for preparing a cannabidiol nano lipid carrier. Background Art

[0002] Cannabidiol (CBD) is a naturally occurring non-psychoactive compound extracted from cannabis. It has multiple biological activities such as antioxidant, antibacterial and anti-inflammatory. In addition, CBD has the effect of repairing damaged skin and improving skin color. Many clinical and preclinical studies have shown that CBD can improve the endogenous cannabinoid system of the human body through multiple biological pathways, thereby effectively controlling the secretion of oil, alleviating acne, and improving skin eczema. However, the skin can effectively prevent the invasion of viruses, bacteria, allergens and toxic chemicals because it is a complex physical barrier, especially the stratum corneum of the skin, which can hinder the penetration of molecules larger than 500Da. Since CBD is a fat-soluble compound, it is prone to photooxidation and has poor water solubility (0.1μg / mL). If it is directly applied to the skin surface, it is easy to form a lipid film, which hinders the breathing of the skin and limits its application in the field of cosmetics and health products. At present, there are many methods to improve the water solubility of fat-soluble compounds, such as nano-microcapsules, polymer nanoparticles, nanocrystals, nanostructured lipid carriers, etc. However, these methods usually have certain disadvantages, such as large particle size (hundreds of nanometers to micrometers), so the transdermal absorption capacity is poor; the particle uniformity is not good, which reduces the physical stability; the preparation process requires the use of organic solvents such as ethanol, which may irritate the skin, etc. Therefore, it is necessary to develop a coating carrier with a simple and environmentally friendly preparation process that can simultaneously improve the water solubility and transdermal absorption capacity of cannabidiol, obtain higher bioavailability, and thus expand its application field. Summary of the invention

[0003] The present invention aims to solve the problem that the existing cannabidiol lipid carriers cannot have both high water solubility and high transdermal absorption function, and have poor stability and are easy to irritate the skin, and further provide a method for preparing a cannabidiol nanolipid carrier with transdermal absorption function.

[0004] A method for preparing a cannabidiol nanolipid carrier with transdermal absorption function is carried out according to the following steps:

[0005] 1. Weighing of raw materials:

[0006] Weigh 0.5% to 2% cannabidiol, 1% to 3% solid oil, 0.4% to 1.2% liquid oil, 1% to 5% emulsifier, 0.2% to 0.8% synergist and the balance aqueous solvent according to mass percentage;

[0007] 2. Preparation of oil phase:

[0008] The weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred evenly to obtain an oil phase;

[0009] 3. Preparation of aqueous phase:

[0010] The weighed emulsifier and aqueous solvent are heated and dissolved, and then stirred evenly to obtain a water phase;

[0011] 4. Mixing:

[0012] The weighed synergist is added to the oil phase at a rate of 10 mL / min to 20 mL / min, and then the water phase is added dropwise at a rate of 5 mL / min to 20 mL / min, and then stirred at a stirring rate of ≥1000 r / min for 20 min to 40 min to obtain colostrum;

[0013] 5. Ultrasonic homogenization:

[0014] Using an ultrasonic cell disruptor, in a water bath temperature of 70°C to 85°C, ultrasonically disperse the colostrum for 3min to 10min to obtain a homogenized mixed solution;

[0015] 6. Cooling:

[0016] The homogenized mixed solution is placed in an ice-water mixture to cool, then taken out and restored to room temperature, and finally filtered to obtain a cannabidiol nano-lipid carrier solution;

[0017] 7. Remove impurities:

[0018] The cannabidiol nano-lipid carrier solution is centrifuged and filtered to remove unencapsulated free cannabidiol, and then redispersed in water to obtain a cannabidiol nano-lipid carrier solution with transdermal absorption function.

[0019] The beneficial effects of the present invention are:

[0020] 1. The preparation method adopted by the present invention is simple in process and time-saving, and the raw materials used are all edible ingredients or natural ingredients, which are non-toxic and biodegradable, and have no irritation to the skin;

[0021] 2. The cannabidiol nano-lipid carrier provided by the present invention can be miscible with water in any proportion, and the average particle size of the particles is small, can be stored for a long time, and has good stability;

[0022] 3. The cannabidiol nano-lipid carrier provided by the present invention can be freeze-dried as needed to increase the convenience of use and storage time;

[0023] 4. The present invention utilizes the terpenes of Ledum palustris that promote transdermal absorption, and together with the coated lipid carrier, forms an efficient penetration-promoting system, which helps cannabidiol to penetrate the stratum corneum, thereby greatly improving the transdermal absorption capacity of cannabidiol;

[0024] 5. The cannabidiol prepared by the present invention is a nano-scale lipid carrier with a controllable particle size between 40nm and 150nm, a strong drug-carrying capacity, and a function of promoting transdermal absorption;

[0025] 6. The cannabidiol nano-lipid carrier prepared by the present invention has an obvious sustained-release effect and can improve the bioavailability of cannabidiol.

[0026] The invention is used for a method for preparing a cannabidiol nano lipid carrier with transdermal absorption function. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The total ion current diagrams of Ledum palustris 1 to Ledum palustris 6 at different distillation times;

[0028] Figure 2 This is an electron micrograph of the cannabidiol nanolipid carrier with transdermal absorption function prepared in Example 5;

[0029] Figure 3 This is an electron micrograph of the cannabidiol nanolipid carrier with transdermal absorption function prepared in Example 8;

[0030] Figure 4 The actual picture of the cannabidiol nanolipid carrier solution with transdermal absorption function prepared in Example 5 diluted with deionized water at different times, a is the cannabidiol nanolipid carrier solution with transdermal absorption function prepared in Example 5, b is diluted 15 times with deionized water, c is diluted 30 times with deionized water, d is diluted 50 times with deionized water, and e is diluted 100 times with deionized water. DETAILED DESCRIPTION

[0031] The technical solution of the present invention is not limited to the specific implementation modes listed below, but also includes any combination of the specific implementation modes.

[0032] Specific embodiment 1: This embodiment is a method for preparing a cannabidiol nanolipid carrier with transdermal absorption function, which is carried out according to the following steps:

[0033] 1. Weighing of raw materials:

[0034] Weigh 0.5% to 2% cannabidiol, 1% to 3% solid oil, 0.4% to 1.2% liquid oil, 1% to 5% emulsifier, 0.2% to 0.8% synergist and the balance aqueous solvent according to mass percentage;

[0035] 2. Preparation of oil phase:

[0036] The weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred evenly to obtain an oil phase;

[0037] 3. Preparation of aqueous phase:

[0038] The weighed emulsifier and aqueous solvent are heated and dissolved, and then stirred evenly to obtain a water phase;

[0039] 4. Mixing:

[0040] The weighed synergist is added to the oil phase at a rate of 10 mL / min to 20 mL / min, and then the water phase is added dropwise at a rate of 5 mL / min to 20 mL / min, and then stirred at a stirring rate of ≥1000 r / min for 20 min to 40 min to obtain colostrum;

[0041] 5. Ultrasonic homogenization:

[0042] Using an ultrasonic cell disruptor, in a water bath temperature of 70°C to 85°C, ultrasonically disperse the colostrum for 3min to 10min to obtain a homogenized mixed solution;

[0043] 6. Cooling:

[0044] The homogenized mixed solution is placed in an ice-water mixture to cool, then taken out and restored to room temperature, and finally filtered to obtain a cannabidiol nano-lipid carrier solution;

[0045] 7. Remove impurities:

[0046] The cannabidiol nano-lipid carrier solution is centrifuged and filtered to remove unencapsulated free cannabidiol, and then redispersed in water to obtain a cannabidiol nano-lipid carrier solution with transdermal absorption function.

[0047] The beneficial effects of this embodiment are:

[0048] 1. The preparation method adopted in this embodiment is simple in process and time-saving, and the raw materials used are all edible ingredients or natural ingredients, which are non-toxic and biodegradable, and have no irritation to the skin;

[0049] 2. The cannabidiol nano-lipid carrier provided in this embodiment can be miscible with water in any proportion, and the average particle size of the particles is small, can be stored for a long time, and has good stability;

[0050] 3. The cannabidiol nano-lipid carrier provided in this embodiment can be freeze-dried as needed to increase the convenience of use and storage time;

[0051] 4. This embodiment utilizes the Ledum terpene that promotes transdermal absorption, and together with the coated lipid carrier, forms an efficient penetration-promoting system to help cannabidiol penetrate the stratum corneum, greatly improving the transdermal absorption capacity of cannabidiol;

[0052] 5. The cannabidiol prepared in this embodiment is a nano-scale lipid carrier with a controllable particle size between 40nm and 150nm, a strong drug-carrying capacity, and a function of promoting transdermal absorption;

[0053] 6. The cannabidiol nano-lipid carrier prepared in this embodiment has an obvious sustained-release effect and can improve the bioavailability of cannabidiol.

[0054] Specific implementation method 2: This implementation method is different from specific implementation method 1 in that: the purity of cannabidiol described in step 1 is greater than 99%; the solid oil described in step 1 is one or a combination of monostearate glyceryl, laurate glyceryl, distearate glyceryl and palmitate glyceryl; the liquid oil ester described in step 1 is one or a combination of linseed oil, squalane, palm oil, hemp oil, olive oil, vitamin E, tea seed oil, almond oil, jojoba oil, grape seed oil, peppermint oil and eucalyptus oil; the emulsifier described in step 1 is a combination of two or more of Tween series, poloxamer series and phospholipids; the aqueous solvent described in step 1 is deionized water or a buffer solvent, the buffer solvent is a phosphate buffer solution, disodium hydrogen phosphate-citric acid buffer or glycine buffer, and the pH of the buffer solvent is 6.5-8.0. Others are the same as specific implementation method 1.

[0055] Specific embodiment 3: This embodiment is different from specific embodiment 1 or 2 in that: the synergist described in step 1 is one or a combination of several of the different components of Ledum terpenes. The rest is the same as specific embodiment 1 or 2.

[0056] Specific embodiment 4: This embodiment is different from the specific embodiments 1 to 3 in that: the Ledum terpenes containing different components are specifically prepared according to the following steps:

[0057] Fresh Ledum odoratum flowers and young leaves in spring and summer are collected, washed, ventilated and dried in the shade to obtain pretreated Ledum odoratum flowers and young leaves; the pretreated Ledum odoratum flowers and young leaves are placed in a steam distillation device, and then an auxiliary extracting liquid is added, and the Ledum odoratum flowers and young leaves are heated to 30°C to 35°C under the condition of a stirring speed of 50 r / min to 70 r / min, and the temperature is stirred at a constant temperature of 30°C to 35°C for 3h to 5h, and then the temperature is raised to 105°C to 110°C under the condition of a stirring speed of 250 r / min to 350 r / min and a heating rate of 7°C / min to 9°C / min, and the temperature is 105°C to 110°C. Under the condition of 0°C, the distillation time is calculated from the distillation of the first drop of essential oil, and 6 kinds of Ledum terpenes with different components are separated from the distillation of the first drop of essential oil to 15 minutes, 16 minutes to 30 minutes, 31 minutes to 60 minutes, 61 minutes to 90 minutes, 91 minutes to 120 minutes, and 121 minutes to 240 minutes; the mass ratio of the pretreated Ledum flowers and young leaves to the auxiliary extract is 1: (10-25); the auxiliary extract is composed of 2% to 5% sodium chloride, 1% to 2% sodium bicarbonate and the balance water in mass percentage. The rest is the same as the specific embodiments one to three.

[0058] Specific embodiment 5: This embodiment is different from specific embodiments 1 to 4 in that: in step 2, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved at a temperature of 70°C to 85°C, and then stirred at a stirring speed of 500r / min to 800r / min for 10min to 25min to obtain an oil phase. The rest is the same as specific embodiments 1 to 5.

[0059] Specific embodiment 6: This embodiment is different from specific embodiments 1 to 5 in that: in step 3, the weighed emulsifier and aqueous solvent are heated and dissolved at a temperature of 70°C to 85°C, and then stirred at a stirring speed of 500r / min to 800r / min for 20min to 30min to obtain an aqueous phase. The rest is the same as specific embodiments 1 to 5.

[0060] Specific embodiment 7: This embodiment is different from specific embodiments 1 to 6 in that the ultrasonic dispersion described in step 5 is specifically carried out according to the following steps: ultrasonic dispersion is carried out for 3 min to 10 min under the conditions of a water bath temperature of 70°C to 85°C and an amplitude of 40% to 70%, and ultrasonic dispersion is carried out for 4s to 10s and then stopped for 4s to 12s. The rest is the same as specific embodiments 1 to 6.

[0061] Specific embodiment 8: This embodiment differs from specific embodiments 1 to 7 in that the filtration described in step 6 is performed using a 0.22 μm filter membrane. The rest is the same as specific embodiments 1 to 7.

[0062] Specific embodiment 9: This embodiment differs from any one of specific embodiments 1 to 8 in that the centrifugal filtration described in step 7 is specifically performed according to the following steps:

[0063] The cannabidiol nano-lipid carrier solution is placed in a centrifugal filtration ultrafiltration tube, and centrifuged for 8 to 15 minutes at a speed of 3000 r / min to 6000 r / min to remove the uncoated free cannabidiol, and then the centrifuged cannabidiol nano-lipid carrier is dispersed in water again, and the centrifugation is repeated 2 to 3 times; the concentration of the cannabidiol nano-lipid carrier solution with transdermal absorption function described in step 7 is 4.5 mg / mL to 18 mg / mL. The rest is the same as the specific embodiments 1 to 8.

[0064] Specific embodiment 10: This embodiment differs from Specific embodiments 1 to 9 in that: in step 7, the cannabidiol nanolipid carrier solution with transdermal absorption function is freeze-dried; the freeze-drying is specifically performed under the conditions of a cold trap temperature of -70°C to -80°C and a vacuum degree of less than 0.1 Torr, and cooling for 24h to 48h. The rest is the same as Specific embodiments 1 to 9.

[0065] The following examples are used to verify the beneficial effects of the present invention:

[0066] The Ledum terpenes containing different components described in the following examples are specifically prepared according to the following steps:

[0067] Fresh Ledum odoratum flowers and young leaves in spring and summer are collected, washed, ventilated and dried in the shade to obtain pretreated Ledum odoratum flowers and young leaves; the pretreated Ledum odoratum flowers and young leaves are placed in a steam distillation device, and then an auxiliary extracting liquid is added, and the mixture is heated to 35°C at a stirring speed of 70 r / min, and stirred at a constant temperature of 35°C for 4 hours, and then heated to 110°C at a stirring speed of 300 r / min and a heating rate of 8°C / min, and the distillation time is calculated from the distillation of the first drop of essential oil at a temperature of 110°C. The first drop of essential oil is distilled for 15 minutes, 16 minutes to 30 minutes, 31 minutes to 60 minutes, 61 minutes to 90 minutes, 91 minutes to 120 minutes, and 121 minutes to 240 minutes to separate and obtain 6 kinds of Ledum terpenes with different components, which are named Ledum 1 to Ledum 6 respectively; the mass ratio of the pretreated Ledum flowers and young leaves to the auxiliary extract is 1:15; the auxiliary extract consists of 3.5% sodium chloride, 1.8% sodium bicarbonate and the remainder water in mass percentage.

[0068] Figure 1 Figure 3 is the total ion current diagram of Ledum palustris 1 to Ledum palustris 6 at different distillation times; it can be seen from the figure that the essential oil fractions obtained at different times are quite different. The Ledum palustris terpenes obtained in the first 60 minutes are mainly 10-carbon monoterpenes and oxygen-containing monoterpenes, and the main components are cymene, pinolene alcohol, α-pinolene, α-thuja aldehyde, myrtle aldehyde, etc. After 90 minutes, the content of oxygen-containing monoterpenes increases, and sesquiterpenes and oxygen-containing sesquiterpenes appear, such as terpenol and terpenol alcohol.

[0069] Embodiment 1:

[0070] A method for preparing a cannabidiol nanolipid carrier with transdermal absorption function is carried out according to the following steps:

[0071] 1. Weighing of raw materials:

[0072] Weigh 1.6% cannabidiol, 2% solid oil, 0.8% liquid oil, 3.2% emulsifier, 0.6% synergist and the balance aqueous solvent according to mass percentage;

[0073] The purity of the cannabidiol is >99%;

[0074] The solid oil is glyceryl monostearate;

[0075] The liquid oil ester is almond oil;

[0076] The emulsifier is a mixture of Tween 80 and soybean lecithin, and the mass ratio of Tween 80 to soybean lecithin is 4:1;

[0077] The aqueous solvent is disodium hydrogen phosphate-citric acid buffer; the pH of the buffer solvent is 7.5;

[0078] The synergist is a mixture of Ledum palustris 1 and Ledum palustris 2, and the mass ratio of Ledum palustris 1 to Ledum palustris 2 is 1:1;

[0079] 2. Preparation of oil phase:

[0080] Under the condition of temperature of 83°C, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred at a stirring speed of 600 r / min for 15 minutes to obtain an oil phase;

[0081] 3. Preparation of aqueous phase:

[0082] The weighed emulsifier and aqueous solvent were heated and dissolved at 83°C, and then stirred at a stirring speed of 600 r / min for 20 min to obtain a water phase;

[0083] 4. Mixing:

[0084] The weighed synergist was added to the oil phase at a rate of 10 mL / min, and then the water phase was added dropwise at a rate of 10 mL / min, and then stirred for 30 minutes at a stirring rate of 1000 r / min to obtain colostrum;

[0085] 5. Ultrasonic homogenization:

[0086] Using an ultrasonic cell disruptor, at a water bath temperature of 80°C and an amplitude of 40%, the colostrum was ultrasonically dispersed for 6 minutes, and ultrasonication was stopped for 4 seconds every 6 seconds to obtain a homogenized mixed solution;

[0087] 6. Cooling:

[0088] The homogenized mixed solution was placed in an ice-water mixture to cool, then taken out and restored to room temperature, and finally filtered using a 0.22 μm filter membrane to obtain a cannabidiol nano-lipid carrier solution;

[0089] 7. Remove impurities:

[0090] The cannabidiol nano-lipid carrier solution is placed in a centrifugal ultrafiltration tube, and centrifuged at a speed of 4000 r / min for 10 minutes to remove uncoated free cannabidiol, and then the cannabidiol nano-lipid carrier separated by centrifugation is dispersed in water again, and the centrifugation is repeated twice, and then dispersed in water again to obtain a cannabidiol nano-lipid carrier solution with transdermal absorption function;

[0091] The concentration of the cannabidiol nanolipid carrier solution with transdermal absorption function is 13.7 mg / mL.

[0092] Example 2: This example is different from Example 1 in that in step 5, an ultrasonic cell disruptor is used to ultrasonically disperse the colostrum for 6 minutes at a water bath temperature of 83° C. and an amplitude of 70%, and the ultrasonication is stopped for 8 seconds after each 4 seconds. The rest is the same as Example 1.

[0093] Embodiment three:

[0094] A method for preparing a cannabidiol nanolipid carrier with transdermal absorption function is carried out according to the following steps:

[0095] 1. Weighing of raw materials:

[0096] Weigh 1.5% cannabidiol, 1.9% solid oil, 0.8% liquid oil, 4% emulsifier, 0.4% synergist and the balance aqueous solvent according to mass percentage;

[0097] The purity of the cannabidiol is >99%;

[0098] The solid oil is a mixture of lauric acid glyceryl and monostearate glyceryl, and the mass ratio of lauric acid glyceryl to monostearate glyceryl is 3:1;

[0099] The liquid oil ester is linseed oil;

[0100] The emulsifier is a mixture of Tween 20 and Poloxamer 188, and the mass ratio of Tween 20 to Poloxamer 188 is 1:1;

[0101] The aqueous solvent is deionized water;

[0102] The synergist is Ledum palustris 5;

[0103] 2. Preparation of oil phase:

[0104] Under the condition of temperature of 80°C, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred at a stirring speed of 500 r / min for 25 minutes to obtain an oil phase;

[0105] 3. Preparation of aqueous phase:

[0106] Under the condition of temperature of 83°C, the weighed emulsifier and aqueous solvent were heated to dissolve, and then stirred at a stirring speed of 800 r / min for 30 min to obtain a water phase;

[0107] 4. Mixing:

[0108] The weighed synergist was added to the oil phase at a rate of 15 mL / min, and then the water phase was added dropwise at a rate of 8 mL / min, and then stirred for 40 minutes at a stirring rate of 1000 r / min to obtain colostrum;

[0109] 5. Ultrasonic homogenization:

[0110] Using an ultrasonic cell disruptor, at a water bath temperature of 80°C and an amplitude of 60%, the colostrum was ultrasonically dispersed for 4 minutes, with ultrasonication for 5 seconds and then stopped for 5 seconds, to obtain a homogenized mixed solution;

[0111] Steps 6 and 7 are the same as those in Example 1, and a cannabidiol nanolipid carrier solution with transdermal absorption function is obtained.

[0112] Embodiment 4:

[0113] A method for preparing a cannabidiol nanolipid carrier with transdermal absorption function is carried out according to the following steps:

[0114] 1. Weighing of raw materials:

[0115] Weigh 1.5% cannabidiol, 1.9% solid oil, 0.8% liquid oil, 4% emulsifier, 0.6% synergist and the balance aqueous solvent according to mass percentage;

[0116] The purity of the cannabidiol is >99%;

[0117] The solid oil is a mixture of lauric acid glyceryl and monostearate glyceryl, and the mass ratio of lauric acid glyceryl to monostearate glyceryl is 2:1;

[0118] The liquid oil ester is linseed oil;

[0119] The emulsifier is a mixture of Tween 20 and Poloxamer 188, and the mass ratio of Tween 20 to Poloxamer 188 is 1:3;

[0120] The aqueous solvent is deionized water;

[0121] The synergist is a combination of Ledum palustris 1, Ledum palustris 3 and Ledum palustris 4, and the mass ratio of Ledum palustris 1:Ledum palustris 3:Ledum palustris 4 is 1:1:1;

[0122] 2. Preparation of oil phase:

[0123] Under the condition of temperature of 78°C, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred at a stirring speed of 800 r / min for 15 minutes to obtain an oil phase;

[0124] 3. Preparation of aqueous phase:

[0125] Under the condition of temperature of 80°C, the weighed emulsifier and aqueous solvent were heated to dissolve, and then stirred at a stirring speed of 700 r / min for 25 min to obtain a water phase;

[0126] 4. Mixing:

[0127] The weighed synergist was added to the oil phase at a rate of 20 mL / min, and then the water phase was added dropwise at a rate of 15 mL / min, and then stirred for 30 minutes at a stirring rate of 1000 r / min to obtain colostrum;

[0128] Steps 5 to 7 are the same as those in Example 1, and a cannabidiol nanolipid carrier solution with transdermal absorption function is obtained.

[0129] Embodiment five:

[0130] A method for preparing a cannabidiol nanolipid carrier with transdermal absorption function is carried out according to the following steps:

[0131] 1. Weighing of raw materials:

[0132] Weigh 1% cannabidiol, 2% solid oil, 1% liquid oil, 3.5% emulsifier, 0.6% synergist and the balance aqueous solvent according to mass percentage;

[0133] The purity of the cannabidiol is >99%;

[0134] The solid oil is a mixture of lauric acid glyceryl and palmitic acid glyceryl, and the mass ratio of lauric acid glyceryl to palmitic acid glyceryl is 1:1;

[0135] The liquid oil ester is a mixture of squalane and palm oil, and the mass ratio of squalane to palm oil is 1:1;

[0136] The emulsifier is a mixture of Tween 80, Poloxamer 188 and egg yolk lecithin, and the mass ratio of Tween 80: Poloxamer 188: egg yolk lecithin is 2:4:1;

[0137] The aqueous solvent is deionized water;

[0138] The synergist is Ledum palustris 3;

[0139] 2. Preparation of oil phase:

[0140] Under the condition of temperature of 72°C, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred at a stirring speed of 600 r / min for 15 minutes to obtain an oil phase;

[0141] 3. Preparation of aqueous phase:

[0142] Under the condition of temperature of 76°C, the weighed emulsifier and aqueous solvent were heated to dissolve, and then stirred at a stirring speed of 700 r / min for 25 min to obtain a water phase;

[0143] 4. Mixing:

[0144] The weighed synergist was added to the oil phase at a rate of 12 mL / min, and then the water phase was added dropwise at a rate of 20 mL / min, and then stirred for 30 minutes at a stirring rate of 1000 r / min to obtain colostrum;

[0145] 5. Ultrasonic homogenization:

[0146] Using an ultrasonic cell disruptor, at a water bath temperature of 80°C and an amplitude of 50%, the colostrum was ultrasonically dispersed for 10 minutes, with ultrasonication for 8 seconds and stopping for 12 seconds, to obtain a homogenized mixed solution;

[0147] Steps 6 and 7 are the same as those in Example 1, and a cannabidiol nanolipid carrier solution with transdermal absorption function is obtained.

[0148] Embodiment six:

[0149] A method for preparing a cannabidiol nanolipid carrier with transdermal absorption function is carried out according to the following steps:

[0150] 1. Weighing of raw materials:

[0151] Weigh 0.5% cannabidiol, 1.5% solid oil, 0.6% liquid oil, 3% emulsifier, 0.4% synergist and the balance aqueous solvent according to mass percentage;

[0152] The purity of the cannabidiol is >99%;

[0153] The solid oil is a mixture of lauric acid glyceryl and palmitic acid glyceryl, and the mass ratio of lauric acid glyceryl to palmitic acid glyceryl is 2:1;

[0154] The liquid oil ester is eucalyptus oil;

[0155] The emulsifier is a mixture of Tween 60, poloxamer 407 and soy lecithin, and the mass ratio of Tween 60: poloxamer 407: soy lecithin is 3:2:1;

[0156] The aqueous solvent is a phosphate buffer solution; the pH of the buffer solvent is 6.5;

[0157] The synergist is Ledum palustris 6;

[0158] 2. Preparation of oil phase:

[0159] Under the condition of temperature of 78°C, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred for 15 minutes at a stirring speed of 700 r / min to obtain an oil phase;

[0160] 3. Preparation of aqueous phase:

[0161] Under the condition of temperature of 82°C, the weighed emulsifier and aqueous solvent were heated to dissolve, and then stirred at a stirring speed of 800 r / min for 30 min to obtain a water phase;

[0162] 4. Mixing:

[0163] The weighed synergist was added to the oil phase at a rate of 20 mL / min, and then the water phase was added dropwise at a rate of 15 mL / min, and then stirred for 30 minutes at a stirring rate of 1000 r / min to obtain colostrum;

[0164] 5. Ultrasonic homogenization:

[0165] Using an ultrasonic cell disruptor, at a water bath temperature of 80°C and an amplitude of 70%, the colostrum was ultrasonically dispersed for 3 minutes, with ultrasonication for 7 seconds and stopping for 5 seconds each time, to obtain a homogenized mixed solution;

[0166] Steps 6 to 7 are the same as those in Example 1, and a cannabidiol nanolipid carrier solution with transdermal absorption function is obtained.

[0167] Embodiment seven:

[0168] A method for preparing a cannabidiol nanolipid carrier with transdermal absorption function is carried out according to the following steps:

[0169] 1. Weighing of raw materials:

[0170] Weigh 1.2% cannabidiol, 1% solid oil, 0.8% liquid oil, 3.3% emulsifier, 0.4% synergist and the balance aqueous solvent according to mass percentage;

[0171] The purity of the cannabidiol is >99%;

[0172] The solid oil is a mixture of lauric acid glyceryl and distearic acid glyceryl, and the mass ratio of lauric acid glyceryl to distearic acid glyceryl is 3:1;

[0173] The liquid oil ester is a mixture of hemp oil and vitamin E, and the mass ratio of hemp oil to vitamin E is 1:1;

[0174] The emulsifier is a mixture of Tween 60, poloxamer 407 and soy lecithin, and the mass ratio of Tween 60: poloxamer 407: soy lecithin is 3:7:1;

[0175] The aqueous solvent is a phosphate buffer solution; the pH of the buffer solvent is 6.5;

[0176] The synergist is Ledum palustris 6;

[0177] 2. Preparation of oil phase:

[0178] Under the condition of temperature of 75°C, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred at a stirring speed of 500 r / min for 10 minutes to obtain an oil phase;

[0179] Steps 3 to 7 are the same as those in Example 1 to obtain cannabidiol nanolipid carriers.

[0180] Embodiment eight:

[0181] A method for preparing a cannabidiol nanolipid carrier with transdermal absorption function is carried out according to the following steps:

[0182] 1. Weighing of raw materials:

[0183] Weigh 0.5% cannabidiol, 2% solid oil, 0.6% liquid oil, 2.2% emulsifier, 0.3% synergist and the balance aqueous solvent according to mass percentage;

[0184] The purity of the cannabidiol is >99%;

[0185] The solid oil is lauric acid glyceride;

[0186] The liquid oil ester is olive oil;

[0187] The emulsifier is a mixture of Tween 20, Poloxamer 188 and egg yolk lecithin, and the mass ratio of Tween 20: Poloxamer 188: egg yolk lecithin is 4:5:1;

[0188] The aqueous solvent is a phosphate buffer solution; the pH of the buffer solvent is 8.0;

[0189] The synergist is Ledum palustris 6;

[0190] 2. Preparation of oil phase:

[0191] Under the condition of temperature of 70°C, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred at a stirring speed of 600 r / min for 15 minutes to obtain an oil phase;

[0192] 3. Preparation of aqueous phase:

[0193] Under the condition of temperature of 70°C, the weighed emulsifier and aqueous solvent were heated to dissolve, and then stirred at a stirring speed of 600 r / min for 25 min to obtain a water phase;

[0194] 4. Mixing:

[0195] The weighed synergist was added to the oil phase at a rate of 20 mL / min, and then the water phase was added dropwise at a rate of 12 mL / min, and then stirred for 30 minutes at a stirring rate of 1000 r / min to obtain colostrum;

[0196] 5. Ultrasonic homogenization:

[0197] Using an ultrasonic cell disruptor, at a water bath temperature of 80°C and an amplitude of 50%, the colostrum was ultrasonically dispersed for 7 minutes, with ultrasonication for 7 seconds and stopping for 5 seconds each time, to obtain a homogenized mixed solution;

[0198] Steps 6 and 7 are the same as those in Example 1, and cannabidiol nanolipid carriers are obtained.

[0199] Comparative experiment: The difference between this embodiment and embodiment 6 is that the weighing of the synergist is omitted in step 1. The rest is the same as embodiment 6.

[0200] (1) Average particle size test:

[0201] Take the cannabidiol nanolipid carrier solution with transdermal absorption function prepared in Examples 1 to 8, dilute it 100 times with deionized water, take 1 mL of the sample after balancing for 15 minutes, and place it in a laser particle size analyzer to measure the average particle size and polydispersity index (PDI) of the sample at 25°C. Each sample is measured three times and the average value is taken. The results are shown in Table 1.

[0202] Table 1 Average particle size and polydispersity coefficient of each example

[0203] sample Average particle size (nm) PDI Embodiment 1 148.7 0.18 Embodiment 2 129.3 0.21 Embodiment 3 103.8 0.17 Embodiment 4 87.6 0.13 Embodiment 5 96.7 0.17 Embodiment 6 58.4 0.13 Embodiment 7 40.3 0.11 Embodiment 8 46.1 0.12

[0204] The results show that the average particle size of the cannabidiol nanolipid carriers with transdermal absorption function prepared in Examples 1 to 8 is less than 150 nm; Figure 2 This is an electron microscope image of the cannabidiol nanolipid carrier with transdermal absorption function prepared in Example 5; as can be seen from the image, the particle size is less than 100 nm; Figure 3 This is an electron microscope image of the cannabidiol nanolipid carrier with transdermal absorption function prepared in Example 8; it can be seen from the figure that the particle size is less than 50nm; and Figure 2 and 3It can be seen that the cannabidiol nanolipid carrier with transdermal absorption function has a regular circular shape. Most of the PDI values ​​are below 0.2, indicating that the system has good dispersibility and a relatively uniform particle size distribution.

[0205] (2) Stability test:

[0206] The cannabidiol nanolipid carrier solutions with transdermal absorption function prepared in Examples 1 to 8 were subjected to stability tests. The average particle size and PDI were measured on the 0th day, 14th day, 30th day, 60th day, 90th day and 180th day of preparation to examine the stability of the lipid carrier. The results are shown in Tables 2 and 3.

[0207] Table 2 Changes of average particle size of each example over time

[0208]

[0209]

[0210] Table 3 PDI changes over time for each example

[0211]

[0212] As can be seen from the table, within 180 days, the average particle size of the cannabidiol nanolipid carriers with transdermal absorption function prepared in Examples 1 to 8 increased compared with the initial preparation, and the PDI also decreased, but the average particle size was still less than 200 nm, and the optimal size was less than 60 nm, indicating that the cannabidiol nanolipid carriers with transdermal absorption function had high stability, were easy to transport and store, and could be applied in the fields of daily chemicals and medicine.

[0213] (3) Water solubility test:

[0214] Figure 4 The actual picture of the cannabidiol nanolipid carrier solution with transdermal absorption function prepared in Example 5 being diluted with deionized water at different times, a is the cannabidiol nanolipid carrier solution with transdermal absorption function prepared in Example 5, b is diluted 15 times with deionized water, c is diluted 30 times with deionized water, d is diluted 50 times with deionized water, and e is diluted 100 times with deionized water; it can be seen from the figure that the cannabidiol nanolipid carrier with transdermal absorption function has good miscibility with water without precipitation, and is miscible with water in any proportion.

[0215] (4) Redissolution test:

[0216] The cannabidiol nanolipid carrier solutions with transdermal absorption function prepared in Examples 1 to 8 were freeze-dried, and the freeze-drying was specifically carried out under the conditions of a cold trap temperature of -80°C and a vacuum degree of 0.09 Torr, and cooled for 48 hours; 0.1 g of the freeze-dried cannabidiol nanolipid carrier powder was mixed with 20 mL of deionized water, and after sufficient shaking, the average particle size and PDI were measured, and the resolubility performance of the cannabidiol nanolipid carrier freeze-dried powder was investigated. The results are shown in Table 4.

[0217] Table 4 Average particle size and polydispersity coefficient after freeze-drying and reconstitution in each embodiment

[0218]

[0219] As can be seen from the table, re-dissolution after freeze-drying will increase the average particle size and PDI of the cannabidiol nanolipid carrier, but the freeze-dried powder can be completely dissolved without precipitation, indicating that the re-dissolution performance of the freeze-dried powder is good.

[0220] (5) In vitro release test:

[0221] The cannabidiol nano lipid carrier solution with transdermal absorption function prepared by embodiment six and the cannabidiol methanol solution with concentration of 60wt% are placed in the phosphate buffer solution (pH 7.0) containing Tween 40 respectively, shaken in a constant temperature shaker (37 ± 1 ℃) for 32h, and 1 ml of supernatant is collected when shaking 0.5, 1, 2, 4, 8, 16 and 32h respectively, and replaced with an equal volume of buffer. Subsequently, the sample is filtered through a 0.2 μm membrane, and the release amount of CBD is measured by HPLC. The cumulative drug release rate Q at each time point is determined by the following calculation, and test result is as shown in Table 5.

[0222]

[0223] Where V0 is the total volume of the release medium, mL; C t is the concentration of cannabidiol at different time points, μg / mL; V is the sampling volume, mL; M0 is the total mass of cannabidiol, mg.

[0224] Table 5 Cumulative release rate of cannabidiol in nanolipid carrier and in free state in Example 6

[0225]

[0226]

[0227] It can be seen from the table that compared with the free state, the cannabidiol nanolipid carrier has an obvious sustained-release effect, which can significantly prolong the action time of cannabidiol and improve its bioavailability.

[0228] (6) Transdermal absorption test:

[0229] The isolated mouse abdominal skin was fixed on the diffusion interface of the diffusion cell (receiving cell volume 5 mL, effective transdermal area 0.65 cm) with the stratum corneum facing upwards using the Franz diffusion cell method. 2 ), a phosphate buffer solution (pH 6.8) of poloxamer 188 (2%, w / v) was added to the receiving cell to allow the receiving solution to fully contact the inner surface of the skin, and the water bath was circulated (37.0±0.5°C). The sample solution was the undiluted cannabidiol nanolipid carrier solution with transdermal absorption function prepared in Example 6, the comparative experiment was the cannabidiol nanolipid carrier solution obtained by the same preparation method but without adding a synergist, and the reference solution was a 60% methanol solution of cannabidiol.

[0230] Add 2.0mL of sample solution or reference solution to the drug supply pool, stir at a constant speed of 200r / min, take 1.0mL of receiving solution at 0, 2, 4, 6, 8, 12, and 24h, and add an equal volume of receiving solution at the same temperature and remove bubbles. After the receiving solution is dried at low temperature, it is re-dissolved with methanol, centrifuged and the supernatant is taken. The cannabidiol content is determined by HPLC, and the cumulative drug permeation per unit area Q is calculated according to the formula 累积 And the transdermal percentage W, calculated by the formula:

[0231]

[0232] Where: V is the volume of the receiving pool, mL; V S is the sampling volume, mL; C n is the drug mass concentration in the receiving solution measured at the nth sampling point, μg / mL; C n-1 is the drug mass concentration in the receiving solution measured at n-1 sampling points, μg / mL; A is the effective transdermal absorption area, cm 2 ;Q 24 The cumulative drug permeation in 24 hours, μg cm -2 ; V c is the total volume of the receiving solution, mL; M is the content of active matter in the upper layer sample of the receiving pool, μg.

[0233] (7) Intradermal retention test:

[0234] At the end of transdermal absorption, the mouse skin was completely removed, cleaned with saline and ethanol, dried and crushed, and ultrasonically extracted with 2 mL of anhydrous ethanol for 10 min. This process was repeated three times, and the extracts were combined, filtered and fixed to 10 mL. The cannabidiol content was determined by HPLC, and the skin retention amount M was determined. C And the absorption percentage P, calculated as:

[0235]

[0236] Where C 24 is the sample concentration at the 24th hour sampling point, μg / mL; T is the dilution factor; A is the effective skin penetration area, cm 2 ; M is the content of active matter in the upper layer sample of the receiving pool, μg.

[0237] The results of the percutaneous absorption test are shown in Table 5.

[0238] Table 6 Results of transdermal release and intradermal retention of samples and reference substances

[0239]

[0240] Note: - means not detected

[0241] As can be seen from the table, cannabidiol itself has poor percutaneous permeability, and the stratum corneum of the skin has a huge hindering effect on it. However, when cannabidiol is coated with the nanostructured liposomes of the embodiment, the transdermal absorption capacity is greatly enhanced. This is because the cannabidiol nanolipid carrier is smaller in size, easier to penetrate the skin barrier, and after coating, a similar skin surface lipid structure is formed, which is easy to absorb. The added synergist Dulemon terpenes can promote the expansion of the intercellular space of the epidermis, which is beneficial for the diffusion of cannabidiol through the intercellular space of the epidermis.

Claims

1. A preparation method of cannabidiol nanolipid carrier with transdermal absorption function, characterized in that It is carried out according to the following steps:

1. Weighing of raw materials: Weigh 0.5% to 2% cannabidiol, 1% to 3% solid oil, 0.4% to 1.2% liquid oil, 1% to 5% emulsifier, 0.2% to 0.8% synergist and the balance aqueous solvent according to mass percentage; 2. Preparation of oil phase: The weighed cannabidiol, solid oil and liquid oil are heated and dissolved, and then stirred evenly to obtain an oil phase; 3. Preparation of aqueous phase: The weighed emulsifier and aqueous solvent are heated and dissolved, and then stirred evenly to obtain a water phase; 4. Mixing: The weighed synergist is added to the oil phase at a rate of 10 mL / min to 20 mL / min, and then the water phase is added dropwise at a rate of 5 mL / min to 20 mL / min, and then stirred at a stirring rate of ≥1000 r / min for 20 min to 40 min to obtain colostrum; 5. Ultrasonic homogenization: Using an ultrasonic cell disruptor, under the condition of a water bath temperature of 70°C to 85°C, ultrasonically disperse the colostrum for 3min to 10min to obtain a homogenized mixed solution; 6. Cooling: The homogenized mixed solution is placed in an ice-water mixture to cool, then taken out and restored to room temperature, and finally filtered to obtain a cannabidiol nano-lipid carrier solution; 7. Remove impurities: The cannabidiol nano-lipid carrier solution is centrifuged and filtered to remove unencapsulated free cannabidiol, and then redispersed in water to obtain a cannabidiol nano-lipid carrier solution with transdermal absorption function.

2. The method for preparing a cannabidiol nano lipid carrier with transdermal absorption function according to claim 1, characterized in that The purity of cannabidiol described in step one is greater than 99%; the solid oil described in step one is one of monostearate glyceryl, laurate glyceryl, distearate glyceryl and palmitate glyceryl or a combination of several thereof; the liquid oil ester described in step one is one of linseed oil, squalane, palm oil, hemp oil, olive oil, vitamin E, tea seed oil, almond oil, jojoba oil, grape seed oil, peppermint oil and eucalyptus oil or a combination of several thereof; the emulsifier described in step one is a combination of two or more of Tween series, poloxamer series and phospholipids; the aqueous solvent described in step one is deionized water or a buffer solvent, the buffer solvent is a phosphate buffer solution, disodium hydrogen phosphate-citrate buffer or glycine buffer, and the pH of the buffer solvent is 6.5-8.

0.

3. The method for preparing a cannabidiol nano lipid carrier with transdermal absorption function according to claim 1, characterized in that The synergist described in step 1 is one or a combination of several of the Ledum terpenes containing different components.

4. The method for preparing a cannabidiol nano lipid carrier with transdermal absorption function according to claim 3, characterized in that The Ledum palustris terpenes containing different components are specifically prepared according to the following steps: Fresh Ledum odoratum flowers and young leaves in spring and summer are collected, washed, ventilated and dried in the shade to obtain pretreated Ledum odoratum flowers and young leaves; the pretreated Ledum odoratum flowers and young leaves are placed in a steam distillation device, and then an auxiliary extracting liquid is added, and the Ledum odoratum flowers and young leaves are heated to 30°C to 35°C under the condition of a stirring speed of 50 r / min to 70 r / min, and the temperature is stirred at a constant temperature of 30°C to 35°C for 3h to 5h, and then the temperature is raised to 105°C to 110°C under the condition of a stirring speed of 250 r / min to 350 r / min and a heating rate of 7°C / min to 9°C / min, and the temperature is 105°C to 110°C. Under the condition of 0°C, the distillation time is calculated from the distillation of the first drop of essential oil, and 6 kinds of Ledum terpenes containing different components are separated from the distillation of the first drop of essential oil to 15 minutes, 16 minutes to 30 minutes, 31 minutes to 60 minutes, 61 minutes to 90 minutes, 91 minutes to 120 minutes, and 121 minutes to 240 minutes respectively; the mass ratio of the pretreated Ledum flowers and young leaves to the auxiliary extract is 1:(10-25); the auxiliary extract consists of 2% to 5% of sodium chloride, 1% to 2% of sodium bicarbonate and the balance water in mass percentage.

5. The method for preparing a cannabidiol nano lipid carrier with transdermal absorption function according to claim 1, characterized in that In step 2, the weighed cannabidiol, solid oil and liquid oil are heated and dissolved at a temperature of 70°C to 85°C, and then stirred at a stirring speed of 500r / min to 800r / min for 10min to 25min to obtain an oil phase.

6. The method for preparing a cannabidiol nano lipid carrier with transdermal absorption function according to claim 1, characterized in that In step 3, the weighed emulsifier and aqueous solvent are heated to dissolve at a temperature of 70° C. to 85° C., and then stirred at a stirring speed of 500 r / min to 800 r / min for 20 min to 30 min to obtain an aqueous phase.

7. The method for preparing a cannabidiol nano lipid carrier with transdermal absorption function according to claim 1, characterized in that The ultrasonic dispersion described in step 5 is specifically carried out according to the following steps: ultrasonic dispersion for 3 minutes to 10 minutes under the conditions of a water bath temperature of 70°C to 85°C and an amplitude of 40% to 70%, and ultrasonic dispersion for 4 seconds to 10 seconds and stop ultrasonication for 4 seconds to 12 seconds.

8. The method for preparing a cannabidiol nano lipid carrier with transdermal absorption function according to claim 1, characterized in that The filtration described in step six is ​​performed using a 0.22 μm filter membrane.

9. The method for preparing a cannabidiol nanolipid carrier with transdermal absorption function according to claim 1, characterized in that The centrifugal filtration described in step 7 is specifically carried out according to the following steps: Place the cannabidiol nanolipid carrier solution in a centrifugal ultrafiltration tube, centrifuge for 8min to 15min at a speed of 3000r / min to 6000r / min to remove uncoated free cannabidiol, then disperse the centrifuged cannabidiol nanolipid carrier into water again, and repeat the centrifugation 2 to 3 times; the concentration of the cannabidiol nanolipid carrier solution with transdermal absorption function described in step 7 is 4.5mg / mL to 18mg / mL.

10. The method for preparing a cannabidiol nano lipid carrier with transdermal absorption function according to claim 1, characterized in that In step seven, the cannabidiol nanolipid carrier solution with transdermal absorption function is freeze-dried; the freeze-drying is specifically carried out under the conditions of a cold trap temperature of -70°C to -80°C and a vacuum degree of less than 0.1 Torr, and cooling for 24h to 48h.

Images