Preparation method of s-(-)-nicotine nanoflexible liposome hydrogel patch

Abstract

The application provides a preparation method of S-(-)-nicotine nanoflexible liposome hydrogel patches, which comprises the following steps: 1) preparing S-(-)-nicotine-loaded nanoflexible liposomes by using a film dispersion method; 2) uniformly mixing the S-(-)-nicotine-loaded nanoflexible liposomes obtained in the step 1) with an aqueous phase matrix to obtain a mixed solution; 3) adding the mixed solution obtained in the step 2) into a hydrogel matrix to obtain a drug-loaded matrix; 4) uniformly coating the drug-loaded matrix obtained in the step 3) on a non-woven fabric, drying and solidifying, covering a protective layer, and obtaining the S-(-)-nicotine nanoflexible liposome hydrogel patch. The S-(-)-nicotine nanoflexible liposome hydrogel patch provided by the application can release S-(-)-nicotine for a long time, has good skin compliance, high adhesion performance and relatively low cost, and has a longer drug release time compared with traditional nicotine patches, so that the use frequency of users can be reduced and the compliance can be increased.

Description

Technical Field

[0001] This invention patent relates to the field of pharmaceutical formulation technology, and in particular to a method for preparing an S-(-)-nicotine nanoflexible liposome hydrogel patch (S-(-)-Nicotine-NFL-HGP). Background Technology

[0002] Nicotine withdrawal syndrome refers to the symptoms experienced by long-term, heavy smokers who, after quitting, experience a lack of nicotine in their bodies, leading to circulatory and endocrine disorders, resulting in physical discomfort and adverse reactions such as irritability and restlessness. In severe cases, joint pain and gastrointestinal discomfort may occur. Quitting smoking without assistance is extremely difficult; studies have found that the failure rate of unassisted quitting is as high as 90-95%, while effective drug interventions can improve the success rate. Nicotine replacement therapy (NRT), as a treatment for nicotine withdrawal syndrome, works by alleviating withdrawal symptoms through a short-term reward mechanism, gradually reducing the smoker's craving for nicotine and ultimately enabling complete elimination of dependence. A specific amount of nicotine is absorbed transdermally into the bloodstream and eventually reaches the brain, producing a feeling of satisfaction. Numerous clinical trials and long-term use results indicate that NRT can improve the success rate of quitting smoking. Nicotine patches, as a form of NRT, are applied directly to the skin and deliver nicotine at a relatively high rate. Patches come in a range of nicotine doses that allow smokers to gradually reduce their nicotine intake, enabling their bodies to gradually adapt to lower nicotine levels and eventually achieve a nicotine-free state.

[0003] The main advantage of patches compared to other formulations is their ease of use; patients simply apply the nicotine patch to their skin, rather than actively using the product all day. In addition, patches offer a less unpleasant sensory experience and deliver nicotine more slowly than other formulations.

[0004] Based on the advantages of nicotine patches as a form of nicotine replacement therapy for treating nicotine withdrawal syndrome, and the advantages of nano-flexible liposome hydrogel drug delivery systems, the development of a nicotine patch that can release nicotine for a long time, has good skin compliance, high adhesion, and relatively low cost is a problem that needs to be solved. Summary of the Invention

[0005] During the research process, the inventors discovered that the nano-flexible liposome hydrogel patch is made by adding a suspension of drug-loaded nano-flexible liposomes into a portion of the water-soluble polymer materials that make up the hydrogel patch, and then solidifying it to form a paste-like patch with a certain viscosity. The combination of the two has certain complementary and synergistic advantages, specifically manifested in the following aspects: (1) The hydrogel matrix not only has good compatibility with the nano-flexible liposomes, but also improves the stability of the flexible nano-liposomes, solving the problem that the effective ingredients are easily oxidized and cannot be preserved for a long time during use and storage. (2) The hydrogel system alone has good affinity with the skin, but it cannot meet the viscosity standard and good extensibility required for the patch, including the initial tack and holding power required by the pharmacopoeia standard. After combining with the nano-flexible liposomes, the adhesion ability of the two as a patch to human skin can be further improved. In addition, the dispersant present in the hydrogel can also solve the problem that the nano-flexible liposomes are prone to precipitation after long-term storage. (3) Due to the deformability of flexible liposomes, they meet the characteristics of poultices for use on human skin. At the same time, combined with the good affinity of hydrogels to the skin, the two are better stabilized on the skin surface after combination. The shape of the poultice will not easily bend, wrinkle or deform with the user. (4) Due to the high deformability of flexible liposomes, the moisturizing properties of hydrogels and the effect of transdermal activators in hydrogels, the synergistic effect of the two can effectively promote the drug to enter the epidermis and dermis, realize the sustained release of drugs through the skin, and achieve a long-lasting therapeutic effect. (5) When drugs are encapsulated in the vesicle structure of flexible liposomes and then combined with the multifunctional network structure of hydrogels to form poultices, will the drug release process cause drug loss or staining of clothing?

[0006] The present invention aims to at least partially solve one of the technical problems existing in the prior art. Therefore, in a first aspect, the present invention provides a method for preparing an S-(-)-nicotine nano-flexible liposome hydrogel patch, comprising the following steps:

[0007] 1) Nanoflexible liposomes loaded with S-(-)-nicotine were prepared by thin-film dispersion method;

[0008] 2) Mix the S-(-)-nicotine-loaded flexible liposomes obtained in step 1) with an aqueous matrix to obtain a mixture;

[0009] 3) Add the mixture obtained in step 2) to the hydrogel matrix to obtain the drug-loaded matrix;

[0010] 4) The drug-loaded matrix obtained in step 3) is uniformly coated on the substrate, dried and cured, and covered with a protective layer to obtain the S-(-)-nicotinic acid nano-flexible liposome hydrogel patch.

[0011] In one or more embodiments of the present invention, in step 1), the S-(-)-nicotinic acid-loaded nano-flexible liposomes use phospholipids and cholesterol as phospholipid bilayer framework materials, wherein the phospholipids are selected from at least one of L-α-phosphatidylcholine, phosphatidylethanolamine, phosphatidylcholine, and hydrogenated lecithin; the membrane softener used in the thin film dispersion method is selected from at least one of sorbate, 1,2-propanediol, sodium cholate, sodium deoxycholate, and ethanol.

[0012] Preferably, the S-(-)-nicotinic acid-loaded flexible liposomes use L-α-phosphatidylcholine and cholesterol as the phospholipid bilayer framework materials, 1,2-propanediol as the membrane softener, and the mass ratio of L-α-phosphatidylcholine to cholesterol is (34~38):1; the mass ratio of 1,2-propanediol to L-α-phosphatidylcholine is (8~12):1; and the mass-to-volume ratio of L-α-phosphatidylcholine to S-(-)-nicotinic acid is (1.0~1.5)×10⁻¹⁰. 4 :1.

[0013] Preferably, step 1) further includes the following steps:

[0014] S1. Place L-α-phosphatidylcholine and cholesterol in a reaction vessel, add solvent, sonicate, homogenize, and dry under reduced pressure until a lipid film forms on the wall of the reaction vessel.

[0015] S2. Dissolve S-(-)-nicotine in a first 1,2-propanediol aqueous solution, hydrate it with the lipid film obtained in step S1 under vacuum, add a second 1,2-propanediol aqueous solution, mix and homogenize to obtain S-(-)-nicotine-loaded flexible liposomes.

[0016] Preferably, in step S1, the solvent is selected from C1-C6 alcohols.

[0017] Preferably, in step S2, the hydration temperature is 25~45℃, the hydration time is 1~3h, the first 1,2-propanediol aqueous solution and the second 1,2-propanediol aqueous solution have the same concentration and volume, the concentration of both the first and second 1,2-propanediol aqueous solutions is 1.5-2 g / mL, and the volume ratio of S-(-)-nicotine to the first 1,2-propanediol aqueous solution is (2.5~3.5):1×10 3 .

[0018] In one or more embodiments of the present invention, in step 2), the aqueous matrix comprises 0.16-0.2 parts by weight of tartaric acid, 0.08-0.1 parts by weight of polyvinylpyrrolidone, and 4.8-5.2 parts by weight of water. Preferably, the aqueous matrix comprises 0.18 parts by weight of tartaric acid, 0.09 parts by weight of polyvinylpyrrolidone, and 5 parts by weight of water. The mass-to-volume ratio of the aqueous matrix to the S-(-)-nicotine nano-flexible liposomes is 0.25-0.28 g / mL.

[0019] In one or more embodiments of the present invention, in step 3), the hydrogel matrix is ​​obtained by mixing and stirring sodium polyacrylate, aluminum glycinate, disodium ethylenediaminetetraacetate, azone, glycerol, and 1,2-propanediol. The hydrogel matrix comprises 0.9-1 parts by weight of sodium polyacrylate, 0.12-0.18 parts by weight of aluminum glycinate, 0.12-0.13 parts by weight of disodium ethylenediaminetetraacetate, 0.08-0.1 parts by weight of azone, 3.8-4.2 parts by weight of glycerol, and 1.8-2.2 parts by weight of 1,2-propanediol. Preferably, the hydrogel matrix comprises 0.95 parts by weight of sodium polyacrylate, 0.15 parts by weight of aluminum glycinate, 0.125 parts by weight of disodium ethylenediaminetetraacetate, 0.09 parts by weight of azone, 4 parts by weight of glycerol, and 2 parts by weight of 1,2-propanediol. The mass-to-volume ratio of the hydrogel matrix to the mixture is 0.28-0.32 g / mL.

[0020] In one or more embodiments of the present invention, in step 3), under a first stirring, the mixture obtained in step 2) is added to the hydrogel matrix, and under a second stirring, a drug-loaded matrix is ​​obtained. The first stirring is carried out at 50~80°C and the stirring speed is 300~700 r / min. The stirring time of the second stirring is 10~60 min. The mixture obtained in step 2) is added dropwise to the hydrogel matrix, and the dropping rate is controlled to be 10~30% / min of the total weight of the mixture.

[0021] In one or more embodiments of the present invention, in step 4), the drying and curing temperature is controlled to be 40~60℃ and the drying and curing time is controlled to be 2~12 h.

[0022] In one or more embodiments of the present invention, the method for characterizing the nanoscale properties of S-(-)-Nicotine-NFL and the method for determining the encapsulation efficiency and drug loading include the following:

[0023] (1) The hydration particle size, zeta potential and PDI of diluted S-(-)-Nicotine-NFL were measured using a laser particle size analyzer. The hydration particle size, zeta potential and PDI of each sample were measured three times and the average value was taken.

[0024] (2) After sonicating, diluting and sonicating the freshly prepared S-(-)-Nicotine-NFL suspension, take an appropriate amount of the diluted sample and drop it onto a copper grid. After drying, perform negative staining with phosphotungstic acid, dry again, and place it under a transmission electron microscope for structural observation.

[0025] (3) Prepare standard S-(-)-nicotine aqueous solutions with mass concentrations of 30 μg / mL, 15 μg / mL, 7.5 μg / mL, 3.75 μg / mL, 1.875 μg / mL and 0.938 μg / mL using ultrapure water. After sonication, measure the ultraviolet absorption values ​​of the standard S-(-)-nicotine aqueous solutions at the maximum absorption wavelength λmax=260nm using an ultraviolet spectrophotometer. Plot the concentration of the standard S-(-)-nicotine aqueous solution as the abscissa and the corresponding ultraviolet absorbance at different concentrations as the ordinate to obtain the ultraviolet absorption standard curve of S-(-)-nicotine aqueous solution.

[0026] (4) Take a quantitative amount of S-(-)-Nicotine-NFL suspension into a centrifuge tube, add an equal volume of 10 mg / mL protamine solution and mix rapidly. After high-speed shaking to cause coagulation, centrifuge the sample at 10000 r / min, remove all supernatant, dilute the supernatant with ultrapure water, sonicate the diluted solution again, and measure the UV absorbance of the diluted solution at λmax=260 nm. According to the UV standard curve, calculate the mass concentration of S-(-)-nicotine in the diluted supernatant, and calculate the encapsulation efficiency according to the following formula ( EE ) and drug loading ( DL ):

[0027] %EE = [1- (M0 / M) ]×100%; %DL = [(M×%EE) / M t ]×100%;

[0028] M0 refers to the mass of S-(-)-nicotine in the supernatant, and M refers to the total mass of S-(-)-nicotine. t It refers to the total mass of S-(-)-Nicotine-NFL (including the mass of S-(-)-nicotinic acid loaded in the S-(-)-Nicotine-NFL suspension, as well as the mass of L-α-phosphatidylcholine and cholesterol).

[0029] In one or more embodiments of the present invention, the method for studying the stability of S-(-)-Nicotine-NFL includes the following: determining the particle size, potential and PDI of S-(-)-Nicotine-NFL at different time points (0-60 days) according to the method of step (1) above; determining the encapsulation efficiency of S-(-)-Nicotine-NFL at different time points (0-60 days) according to the method of step (2) above, and calculating the drug leakage rate to evaluate its stability.

[0030] In one or more embodiments of the present invention, the method for detecting the initial tack (IBS) and holding tack (EBS) of S-(-)-Nicotine-NFL-HGP includes the following steps:

[0031] (1) The IBS of the material was determined by the inclined rolling ball method specified in the 2015 edition of the Chinese Pharmacopoeia. The steel ball number, specifications, and HG810 initial tack tester used in the experiment met the requirements. The release paper of the material paste packaging of the test sample patch was slowly peeled off, and the patch was placed at room temperature for 1 hour. The patch was then cut into appropriately sized and identical shapes and placed in the center of the inclined plate of the initial tack tester with the paste side facing up. The inclined plate was set at an angle of 22.5°C to ensure that the patch was fixed in place and did not slip. The upper and lower parts of the inclined plate that were not covered by the patch were covered with a 0.025 mm thick polyester film. Each standard steel ball (steel ball number 1-32) was rolled freely from the top of the inclined plate in descending order until the first steel ball could be stuck to the sample patch (at any position). The steel ball number at this point was recorded. The IBS was represented by the largest steel ball number that at least two identical test sample paste surfaces could stick to. Each test was repeated 3 times, and the average value was taken as the final IBS result.

[0032] (2) The CNY-5 room temperature tack tester conforms to the requirements of the national standard GB / T 4851-2014. Slowly peel off the release paper of the sample patch and place it at room temperature for 1 hour. Wipe the test plate with a cleaning agent and then dry it with a clean gauze. Repeat this process three times or more until the test plate is visually clean. Under certain temperature and humidity conditions, cut the test sample patch into rectangles of the same size, with the width of the rectangles matching the width of the test plate. Then, paste the patch onto the clean test plate, ensuring that the pasting area of ​​each sample is consistent. Use a standard 2 kg pressure roller to press the patch repeatedly three times at a certain speed. Fix the test plate vertically on the test frame and secure the bottom end of the patch connected to the unpainted part of the test plate with a pin. Hang a 1 kg weight on the other end of the pin to start the experiment and start the automatic timing. Record the time (s) until the sample patch completely detaches from the test plate. After the experiment is completed, the same sample is tested 3 times, and the average value of the results is taken as the final EBS result.

[0033] In one or more embodiments of the present invention, the in vitro release experiment of S-(-)-Nicotine-NFL-HGP includes the following steps:

[0034] (1) For the three samples of S-(-)-nicotine hydrogel patch (S-(-)-Nicotine-HGP), S-(-)-Nicotine-NFL-HGP and commercially available Nicorette®, cut a 2 cm × 2 cm area (with a drug loading of 6.96 mg) from the part with relatively uniform coating thickness, peel off the release paper, and attach the gel side to the inner surface of the dialysis bag. At the same time, add a quantitative amount of PBS at pH=7.4 to the dialysis bag. After sealing the dialysis bag, place it in a stoppered conical flask containing a quantitative amount of PBS at pH=7.4 as the release medium, ensuring that the solid part in the dialysis bag is completely immersed in the buffer solution.

[0035] (2) All three samples were placed in a constant temperature shaker at 100 r / min and shaken at 37°C in the dark. At different time points (0~48h), a certain amount of solution was taken from each conical flask and an equal amount of PBS was added. Three control groups were set up for both types of samples. The solutions taken at different time points were numbered, filtered through a 0.45um microporous membrane, and stored in a sealed container at room temperature in the dark.

[0036] (3) After diluting the solutions taken at different time points, the absorbance was measured using a UV spectrophotometer at a wavelength of λmax = 260 nm. The concentration of S-(-)-nicotine was calculated according to the UV standard curve of S-(-)-nicotine. The concentration was multiplied by the corresponding dilution factor to obtain the reagent concentration, and then the cumulative release of S-(-)-nicotine was calculated. M t ) and cumulative release rate ( Q t ).

[0037] M t ; Q t %= × 100%

[0038] V is the volume of each sample taken; c n-1 The mass concentration of S-(-)-nicotine in the solution sampled in the (n-1)th time; V0 is the total volume of the release medium; c n M represents the mass concentration of S-(-)-nicotine in the solution sampled in the nth time; loading drug Drug-loaded hydrogels or Nicorettes ® The mass of S-(-)-nicotine loaded in a 2cm×2cm area.

[0039] In one or more embodiments of the present invention, the preparation method of S-(-)-Nicotine-HGP in step (1) is to directly load the same amount of S-(-)-nicotine as S-(-)-Nicotine-NFL-HGP into the gel matrix, and the preparation method of the gel matrix is ​​the same.

[0040] In one or more embodiments of the present invention, the content of S-(-)-nicotine per unit area of ​​the commercially available Nicorette® patch sample in step (1) is 1.74 mg / cm². 2 The content of S-(-)-nicotine per unit area in S-(-)-Nicotine-HGP and S-(-)-Nicotine-NFL-HGP is based on this standard and remains consistent.

[0041] In one or more embodiments of the present invention, the S-(-)-Nicotine-NFL-HGP transdermal assay includes the following steps:

[0042] (1) Shave the hair off healthy mice, kill them by disembowelment, take the abdominal skin, completely remove the subcutaneous fat, clean it with physiological saline, absorb the surface moisture with filter paper, cut the mouse skin into appropriate size and fix it on the Franz vertical diffusion cell with the stratum corneum facing up.

[0043] (2) The prepared S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, and Nicorette® were cut into patches of the same size and appropriate dimensions. The patches were then attached to the surface of the mouse skin, and air bubbles were removed between the patches and the skin. The patches were fixed between the supply and receiving chambers of the vertical diffusion cell with spring clips. A magnetic stir bar was placed in the receiving chamber, and PBS solution with pH=7.4 was filled in as the receiving solution. Air bubbles were removed. The receiving cell was placed in a 37℃ constant temperature magnetic stirrer with a rotation speed of 800 r / min. Three parallel experiments were set up for each of the three samples, and the average value of the results was taken.

[0044] (3) Samples were taken from the receiving cell at 0-48h and an equal volume of PBS solution was added. The sampled solution was filtered through a 0.45μm microporous membrane. The solutions taken at different time points were numbered and stored in a sealed container at room temperature away from light.

[0045] (4) After diluting the solutions taken at different time points, the absorbance was measured using a UV spectrophotometer at a wavelength of λmax = 260 nm. The concentration of S-(-)-nicotine was calculated according to the UV standard curve of S-(-)-nicotine. The concentration of the released sample was obtained by multiplying the concentration by the corresponding dilution factor, and then the cumulative transmittance of S-(-)-nicotine per unit area was calculated. Q n .

[0046] Q n ) / A

[0047] V is the volume of each sample taken; c n-1 The mass concentration of S-(-)-nicotine in the solution sampled in the (n-1)th time; V0 is the total volume of the release medium; c n denoted as S-(-)-nicotine in the solution sampled for the nth time; A is the effective contact area of ​​the diffusion cell.

[0048] (5) After the transdermal test, the patch was slowly removed, and the isolated rat skin was washed with physiological saline. After drying the skin, it was ground into a paste with a mortar and pestle, dissolved in an appropriate amount of ethanol, and sonicated for 30 min. After centrifugation at 3500 r / min for 20 min, the supernatant was collected, filtered through a 0.22 μm organic membrane, and the S-(-)-nicotinic acid content was determined by the ultraviolet standard curve method. The amount of dermal skin retained per unit area of ​​rat skin was calculated. Each sample group was tested three times, and the average value was taken.

[0049] In one or more embodiments of the present invention, the healthy mice selected in step (1) are almost identical in size and weight, and are raised in the same environment for 24 hours.

[0050] 20. Stability study of S-(-)-Nicotine-NFL-HGP, including the following: Three batches of S-(-)-Nicotine-NFL-HGP samples were sealed in aluminum foil pouches and stored at 4℃. Samples were taken for observation and testing at 0, 15, 30, 45, 60, 75, and 90 days. The appearance, release rate, and tack of the sample patches were compared with those at 0 days. Visual changes of the patch matrix layer were recorded by photographing. Drug release at 48 h was tested using the dialysis method. Tack was tested according to the methods for initial tack and holding power. All results were taken as the average of the three batches of samples.

[0051] The beneficial effects of this invention are as follows:

[0052] 1. The method for preparing S-(-)-Nicotine-NFL-HGP provided by this invention is simple, easy to operate, and requires mild conditions.

[0053] 2. The S-(-)-Nicotine-NFL-HGP provided by this invention can release S-(-)-nicotine in a long-lasting manner, has good skin compliance, high adhesion performance and relatively low cost.

[0054] 3. Compared with traditional nicotine patches, the S-(-)-Nicotine-NFL-HGP provided by this invention has a longer drug release time, which can reduce the frequency of user use and increase compliance. Attached Figure Description

[0055] Figure 1 Image showing the appearance (front of the patch matrix) of S-(-)-Nicotine-NFL-HGP;

[0056] Figure 2 The figure shows the fitting curve of the S-(-)-nicotine UV standard curve;

[0057] Figure 3 Transmission electron microscopy image of S-(-)-Nicotine-NFL;

[0058] Figure 4 The graph shows the changes in the nanoscale properties of S-(-)-Nicotine-NFL at different times: Wherein, Figure 4 A represents the variation trends of particle size and PDI of S-(-)-Nicotine-NFL at different times (0, 10, 20, 30, 40, 50, 60 days). Figure 4 B represents the changing trends of Zeta potential and drug encapsulation efficiency at different time points (0, 10, 20, 30, 40, 50, 60 days).

[0059] Figure 5 This is a trend chart of the stability of S-(-)-Nicotine-NFL-HGP, where, Figure 5 A is a graph showing the initial tack (IBS) and holding tack (EBS) of S-(-)-Nicotine-NFL HGP at different times (0, 15, 30, 45, 60, 75, 90 days); Figure 5 B is a graph showing the drug release rate data of S-(-)-Nicotine-NFL HGP at different time points (0, 15, 30, 45, 60, 75, 90 days); Figure 5 C is a graph showing the appearance changes of S-(-)-Nicotine-NFL HGP at different times (0, 15, 30, 45, 60, 75, 90 days);

[0060] Figure 6 For S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, Nicorette ® In vitro release curves of the three patches;

[0061] Figure 7 For S-(-)-Nicotine-NFL, S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, Nicorette ® Higuchi equation fitting curves for in vitro release of several drug delivery systems, among which... Figure 7 A is the Higuchi equation fitting curve for the in vitro release of S-(-)-Nicotine-NFL; Figure 7 B is the Higuchi equation fitting curve for the in vitro release of S-(-)-Nicotine-HGP; Figure 7 C is the Higuchi equation fitting curve for the in vitro release of S-(-)-Nicotine-NFL-HGP; Figure 7 D is Nicorette ® The fitting curve of the Higuchi equation for in vitro release;

[0062] Figure 8 For S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, Nicorette ® Cumulative transdermal drug release per unit area over 48 hours for three different patches;

[0063] Figure 9 For S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, Nicorette ® Higuchi equation fitting curves for cumulative transdermal drug release per unit area over 48 hours for three patches, where... Figure 9 A is the Higuchi equation fitting curve of cumulative transdermal drug release per unit area of ​​S-(-)-Nicotine-HGP48 h; Figure 9 B is the Higuchi equation fitting curve of cumulative transdermal drug release per unit area of ​​S-(-)-Nicotine-NFL-HGP over 48 h; Figure 9 C is the Higuchi equation fitting curve of cumulative transdermal drug release per unit area of ​​Nicorette® over 48 h;

[0064] Figure 10 For S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, Nicorette ® Bar chart showing the amount of drug retained in the dermis per unit area after 48 hours for three different patches. Detailed Implementation

[0065] The present invention will be further described below with reference to specific embodiments and accompanying drawings. However, the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified, specific conditions in the following embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the methods used are conventional methods known in the art, and the consumables and reagents used are commercially available. Unless otherwise stated, the technical and scientific terms used herein have the same meaning as those familiar to those skilled in the art. Furthermore, any methods or materials similar to or equivalent to those described herein may also be applied to the present invention.

[0066] Examples 1-3 are preparation examples, providing a method for preparing S-(-)-Nicotine-HGP and S-(-)-Nicotine-NFL-HGP (S-(-)-nicotine nano-flexible liposome hydrogel patch).

[0067] Example 1: Preparation of S-(-)-Nicotine-NFL

[0068] S1. Accurately weigh 360 mg of L-α-phosphatidylcholine and 10 mg of cholesterol into a round-bottom flask, add 20 mL of anhydrous ethanol, sonicate for 20 min and homogenize at high speed for 15 min to ensure that the two are evenly dispersed in the ethanol, and then dry under reduced pressure at 45℃ and -0.05 MPa until a dry lipid film forms on the flask wall. Seal and store at room temperature for later use.

[0069] S2. Dissolve 3.6 g of 1,2-propanediol in ultrapure water, bring the volume to 20 mL, and sonicate for 5 min to obtain 20 mL of 1,2-propanediol aqueous solution. Take 10 mL of the sonicated 1,2-propanediol aqueous solution, add 30 μL of S-(-)-nicotine to dissolve, sonicate for 5 min, and then preheat this 10 mL mixed solution at 50 ℃ for 20 min. Slowly add it to the dried lipid membrane mentioned above, and hydrate it with the liposome membrane under vacuum for 120 min.

[0070] S3. After hydration, combine the remaining 10 mL of sonicated 1,2-propanediol aqueous solution with it, mix thoroughly, transfer to an Erlenmeyer flask, homogenize at high speed for 15 min, and obtain about 20 mL of off-white S-(-)-Nicotine-NFL suspension (S-(-)-nicotinic acid-loaded nano-flexible liposomes), and store at 4℃ in a sealed container for later use.

[0071] Example 2: Preparation of S-(-)-Nicotine-NFL-HGP

[0072] S1. Preparation of oil phase (hydrogel matrix): Accurately weigh 950 mg of sodium polyacrylate, 150 mg of aluminum glycinate, 125 mg of disodium EDTA and 90 mg of azone into a mixed solvent consisting of 4 g of glycerol and 2 g of 1,2-propanediol. Stir at 60°C in the dark for 1 h. After mixing evenly, seal and store at 37°C for later use.

[0073] S2. Preparation of aqueous matrix: Weigh 180 mg of tartaric acid and 90 mg of polyvinylpyrrolidone, dissolve them in 5 mL of ultrapure water, and sonicate for 20 min until completely dissolved.

[0074] S3. The above-mentioned aqueous matrix is ​​mixed with 20 mL of the freshly prepared S-(-)-Nicotine-NFL suspension from Example 1. The mixture is ultrasonically dispersed for 10 min to obtain approximately 25 mL of aqueous drug-loaded mixture (drug-loaded matrix). The prepared oil phase is then placed in a 70°C water bath and stirred at 700 r / min. The aqueous drug-loaded mixture (drug-loaded matrix) is added to the oil phase in four batches, with each batch added slowly at a rate of 30% / min, with a 5-min interval between each batch, until all the mixture is added to the oil phase. The mixture is then stirred at 70°C and 500 r / min for 30 min to obtain the matrix material.

[0075] S4. A circular nonwoven fabric with a diameter of 414 mm is used as the backing layer (effective area approximately 13.5 cm²). 2 The aforementioned matrix material was coated onto a nonwoven fabric to form a matrix layer approximately 2 mm thick. Release paper was then placed over the matrix layer, and the mixture was pressed against a glass plate and cured at 50 °C for 4 hours. Complete curing was considered achieved when no residue remained to the touch, yielding S-(-)-Nicotine-NFL-HGP (S-(-)-nicotinic acid nano-flexible liposome hydrogel patch). The prepared S-(-)-Nicotine-NFL-HGP had an area of ​​13.5 cm². 2 The thickness is 2 mm; the total mass containing S-(-)-Nicotine is 23.5 mg, and the drug loading per unit area is 1.74 mg / cm². 2 S-(-)-Nicotine-NFL-HGP appears as a pale yellow, layered paste with a smooth, glossy surface, showing no cold flow or exudation. See attached image for a diagram of the paste surface. Figure 1 .

[0076] Example 3: Preparation of S-(-)-Nicotine-NFL-HGP

[0077] S1. Preparation of oil phase: Accurately weigh 900 mg of sodium polyacrylate, 180 mg of aluminum glycinate, 120 mg of disodium EDTA and 80 mg of azone into a mixed solvent consisting of 4.2 g of glycerol and 2.2 g of 1,2-propanediol. Stir at 60°C in the dark for 1 h. After mixing evenly, seal and store at 37°C for later use.

[0078] S2. Aqueous phase preparation: Weigh 200 mg of tartaric acid and 80 mg of polyvinylpyrrolidone, dissolve them in 5.2 mL of ultrapure water, and sonicate for 20 min until completely dissolved.

[0079] S3. The above-mentioned aqueous phase material is mixed with 20 mL of freshly prepared S-(-)-Nicotine-NFL suspension from Example 1. The mixture is ultrasonically dispersed for 10 min to obtain approximately 25 mL of aqueous drug-loaded mixture. Then, the prepared oil phase is placed in a 50°C water bath and stirred at 500 r / min. The aqueous drug-loaded mixture is added to the oil phase in 4 batches, with each batch added slowly at a rate of 30% / min, with a 5-min interval between each batch, until all the mixture is added to the oil phase. The mixture is then stirred at 70°C and 500 r / min for 10 min to obtain the matrix material.

[0080] S4. A circular nonwoven fabric with a diameter of 414 mm is used as the backing layer (effective area approximately 13.5 cm²). 2 The aforementioned matrix material was coated onto a nonwoven fabric to form a matrix layer approximately 2 mm thick. Release paper was then placed over the matrix layer, and the mixture was pressed against a glass plate and cured at 50 °C for 4 hours. Complete curing was considered achieved when no residue remained to the touch, yielding S-(-)-Nicotine-NFL-HGP. The prepared S-(-)-Nicotine-NFL-HGP had an area of ​​13.5 cm². 2 The thickness is 2 mm; the total mass containing S-(-)-Nicotine is 23.5 mg, and the drug loading per unit area is 1.74 mg / cm². 2 .

[0081] Example 4: Preparation of S-(-)-Nicotine-NFL-HGP

[0082] S1. Oil phase preparation: Accurately weigh 1000 mg of sodium polyacrylate, 120 mg of aluminum glycinate, 130 mg of disodium EDTA and 100 mg of azone, and dissolve them in a mixed solvent consisting of 3.8 g of glycerol and 1.8 g of 1,2-propanediol. Stir at 60°C in the dark for 1 hour. After mixing evenly, seal and store at 37°C for later use.

[0083] S2. Aqueous phase preparation: Weigh 160 mg of tartaric acid and 100 mg of polyvinylpyrrolidone, dissolve them in 4.8 mL of ultrapure water, and sonicate for 20 min until completely dissolved.

[0084] S3. The above-mentioned aqueous phase material is mixed with 20 mL of freshly prepared S-(-)-Nicotine-NFL suspension from Example 1. The mixture is ultrasonically dispersed for 10 min to obtain approximately 25 mL of aqueous drug-loaded mixture. Then, the prepared oil phase is placed in an 80°C water bath and stirred at 300 r / min. The aqueous drug-loaded mixture is added to the oil phase in 4 batches, with each batch added slowly at a rate of 30% / min, with a 5-min interval between each batch, until all the mixture is added to the oil phase. The mixture is then stirred at 70°C and 500 r / min for 60 min to obtain the matrix material.

[0085] S4. A circular nonwoven fabric with a diameter of 414 mm is used as the backing layer (effective area approximately 13.5 cm²). 2 The aforementioned matrix material was coated onto a nonwoven fabric to form a matrix layer approximately 2 mm thick. Release paper was then placed over the matrix layer, and the mixture was pressed against a glass plate and cured at 50 °C for 4 hours. Complete curing was considered achieved when no residue remained to the touch, yielding S-(-)-Nicotine-NFL-HGP. The prepared S-(-)-Nicotine-NFL-HGP had an area of ​​13.5 cm². 2 The thickness is 2 mm; the total mass containing S-(-)-Nicotine is 23.5 mg, and the drug loading per unit area is 1.74 mg / cm². 2 .

[0086] Example 5: Preparation of S-(-)-Nicotine-HGP

[0087] The difference between Example 5 and Example 2 is that S-(-)-nicotine aqueous solution of the same volume and content is used instead of S-(-)-nicotine-NFL in the preparation process, thus obtaining S-(-)-nicotine hydrogel patch (S-(-)-nicotine-HGP).

[0088] Example 6: Nanoscale Characterization of S-(-)-Nicotine-NFL

[0089] S1. Take the prepared S-(-)-Nicotine-NFL suspension, sonicate for 10 min, take out 1 mL, dilute it 5 times with ultrapure water, and then sonicate the diluted sample again for 10 min. Use a laser particle size analyzer to measure the hydrated particle size and Zeta potential of the diluted S-(-)-Nicotine-NFL. The hydrated particle size and Zeta potential of each sample are measured three times, and the average value is taken. The results are shown in Table 1 in S3.

[0090] S2. Construction of the UV Standard Curve: A standard S-(-)-nicotine aqueous solution with a mass concentration of 15 μg / mL was precisely prepared. UV wavelength scanning was performed using a UV spectrophotometer to determine the maximum absorption wavelength λmax of S-(-)-nicotine as 260 nm. Ultrapure water was used to prepare standard S-(-)-nicotine aqueous solutions with mass concentrations of 30, 15, 7.5, 3.75, 1.875, and 0.938 μg / mL. At the maximum absorption wavelength λmax, the UV absorbance values ​​for each concentration were measured using a UV spectrophotometer, and the data were recorded. A linear fit was performed on a scatter plot with concentration on the x-axis and UV absorbance on the y-axis to obtain the UV absorption standard curve: y = 0.0165x + 0.0013, R0. 2 = 0.9992, good fit, see attached UV standard curve. Figure 2 .

[0091] S3. Take V mL of S-(-)-Nicotine-NFL suspension and place it in a centrifuge tube. Add an equal volume of 10 mg / mL protamine solution and mix rapidly. Vibrate at high speed to coagulate for 1 min, then let stand for 5 min. Centrifuge the coagulated sample at 10000 r / min for 15 min. After centrifugation, remove all supernatant. Dilute the supernatant 20 times with ultrapure water. Sonicate V1 mL of the dilution solution for 15 min. Measure the UV absorbance of the dilution solution at λmax = 260 nm using a UV spectrophotometer. Substitute this value into the UV standard curve of S-(-)-Nicotine to calculate the mass concentration (c) of S-(-)-nicotine in the diluted supernatant. Calculate the encapsulation efficiency according to the following formula ( EE ) and drug loading ( DL ).

[0092] %EE = [1-(M0 / M) ]×100%; %DL = [(M×%EE) / M t ]×100%;

[0093] M0 refers to the mass of S-(-)-nicotine in the supernatant, and M refers to the total mass of S-(-)-nicotine. tIt refers to the total mass of S-(-)-nicotinic nanoflexible liposomes (including the mass of S-(-)-nicotinic acid loaded in the S-(-)-Nicotine-NFL suspension, as well as the mass of L-α-phosphatidylcholine and cholesterol).

[0094] Table 1. Nanoscale property characterization results of S-(-)-Nicotine-NFL

[0095]

[0096] S4. After sonicating the 2S-(-)-Nicotine-NFL suspension for 10 min, take out 100 μL and dilute it 20 times with ultrapure water. Then sonicate the diluted sample again for 10 min. Take an appropriate amount of the diluted sample solution and drop it onto a copper grid. After drying, perform negative staining with phosphotungstic acid (1.5%, W / W), dry it again, and observe the structure under a transmission electron microscope. The electron microscope images are attached. Figure 3 .

[0097] Example 7: Stability of S-(-)-Nicotine-NFL

[0098] The research methodology is as follows:

[0099] The particle size, potential, and PDI of S-(-)-Nicotine-NFL at different time points (0-60 days) were determined according to the method in S1 of Example 1; the encapsulation efficiency of S-(-)-Nicotine-NFL at different time points (0-60 days) was determined according to the method in S3 of Example 1, and the drug leakage rate of S-(-)-Nicotine-NFL within two months was calculated to evaluate the stability of S-(-)-Nicotine-NFL. The storage conditions for S-(-)-Nicotine-NFL are: 4°C, sealed and protected from light.

[0100] The experimental results are as follows:

[0101] For liposome nanostructures, particle size, potential, PDI, and drug encapsulation efficiency are the most direct ways to detect whether the nanostructure properties of the material change over a period of time. Table 2 shows that for the S-(-)-Nicotine-NFL prepared in this experiment, under sealed and light-protected storage conditions at 4 °C, the hydrated particle size, PDI, and Zeta potential remained almost unchanged over 60 days, while the drug encapsulation efficiency decreased (78.2%→71.8%), and the drug leakage rate was 8.2%. Therefore, the S-(-)-Nicotine-NFL drug delivery system is prone to drug leakage over long periods and is not suitable as the final drug delivery system for S-(-)-Nicotine. See the attached graph for the trend of nanostructure changes. Figure 4 .in, Figure 4A represents the variation trends of particle size and PDI of S-(-)-Nicotine-NFL at different times (0, 10, 20, 30, 40, 50, 60 days). Figure 4 B represents the changing trends of Zeta potential and drug encapsulation efficiency at different times (0, 10, 20, 30, 40, 50, 60 days).

[0102] Table 2. Nanoscale feature changes of S-(-)-Nicotine-NFL at different time points (n=3)

[0103]

[0104] Example 8: Characterization and Results of Initial Tack (IBS) and Holding Tack (EBS) of S-(-)-Nicotine-NFL-HGP

[0105] The characterization methods are as follows:

[0106] S1. Slowly peel off the release paper from the material packaging of the sample patch and place it at room temperature for 1 hour. Wipe the test board with a cleaning agent and then dry it with a clean gauze. Repeat this process at least three times until the test board is visually clean. Under specific temperature and humidity conditions, cut the test sample patch into rectangles of suitable and identical size, with the width of the rectangle matching the width of the test board. Then, paste the patch onto the clean test board, ensuring that the pasting area of ​​each sample is consistent. Use a standard 2000 g pressure roller to press the patch repeatedly three times at a certain speed. Fix the test board vertically on the test frame and secure the bottom end of the patch connected to the unpainted part of the test board with a pin. Hang a 1 kg weight on the other end of the pin to start the experiment and start the automatic timing. Record the time (s) until the sample patch completely detaches from the test board. After the experiment, repeat the test three times for the same sample and take the average value as the final holding power result.

[0107] S2. Slowly peel off the release paper from the material packaging of the sample patch and leave it at room temperature for 1 hour. Wipe the test board with a cleaning agent and then dry it with a clean gauze. Repeat this process at least three times until the test board is visually clean. Under specific temperature and humidity conditions, cut the test sample patch into rectangles of suitable and identical size, with the width of the rectangle matching the width of the test board. Then, paste the patch onto the clean test board, ensuring consistent pasting area for each sample. Use a standard 2000 g pressure roller to press the patch repeatedly three times at a constant speed. Fix the test board vertically to the test frame, securing one end of the pin to the unpainted portion of the patch on the test board. Hang a 1 kg weight on the other end of the pin to start the experiment, and start the automatic timing. Record the time (s) until the sample patch completely detaches from the test board. Repeat the experiment three times for the same sample, and take the average result as the final holding power result.

[0108] The results are as follows:

[0109] Under optimal preparation conditions, the initial tack of the S-(-)-Nicotine-NFL-HGP patch is 17±1 (standard steel ball number), and the holding power is (27.1±1.6) s, which meets the standard for long-term adhesion to human skin.

[0110] Stability of Example 9S-(-)-Nicotine-NFL-HGP

[0111] Research Methods:

[0112] Three batches of S-(-)-Nicotine-NFL-HGP samples were sealed in aluminum foil pouches and stored at 4°C. Samples were taken at 0, 15, 30, 45, 60, 75, and 90 days for observation and testing. The appearance, release rate, and tack of the sample patches were compared with those at 0 days. Visual changes of the patch matrix layer were recorded by photographing. The 48-hour drug release rate was tested using the in vitro release dialysis method described in Example 7. The tack was tested using the initial tack and holding power testing methods described in S1 and S2 of Example 5.

[0113] Experimental results:

[0114] Initial tack and holding power did not change significantly over 90 days, with the initial tack remaining between 15 and 20 (standard steel ball size). This indicates that the experimentally prepared S-(-)-Nicotine-NFL-HGP patch has good viscosity stability. Drug release decreased slightly from (64.8±2.5)% to (59.4±4.6)% over 90 days, indicating stable drug release performance of the patch. Furthermore, the appearance changes of the patch over 90 days are shown; the appearance of S-(-)-Nicotine-NFL HGP remained almost unchanged, consistently a light yellow color. Specific experimental data and appearance changes are shown in the appendix. Figure 5 .in, Figure 5 A is a graph showing the initial tack (IBS) and holding tack (EBS) of S-(-)-Nicotine-NFL HGP at different times (0, 15, 30, 45, 60, 75, 90 days); Figure 5 B is a graph showing the drug release rate data of S-(-)-Nicotine-NFL HGP at different time points (0, 15, 30, 45, 60, 75, 90 days); Figure 5 C represents the appearance changes of S-(-)-Nicotine-NFL HGP at different times (0, 15, 30, 45, 60, 75, 90 days).

[0115] Example 10: In vitro release of S-(-)-Nicotine-NFL-HGP

[0116] The research methodology is as follows:

[0117] Preparation of S1, pH=7.4 PBS: Accurately weigh 8 g NaCl, 0.2 g KCl, 1.44 g Na2HPO4 and 0.24 g KH2PO4 and dissolve them in 800 mL of ultrapure water. Adjust the pH of the solution to 7.4 with dilute hydrochloric acid, sonicate for 20 min, and store at room temperature for later use.

[0118] S2. For the prepared S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, and Nicorette®, cut off a 2cm × 2cm area (with a drug loading of 6.96 mg) from the area with relatively uniform coating thickness. Slowly peel off the release paper from the cut-off portion and place the gel side onto the inner surface of a dialysis bag with a molecular weight cutoff of 8000~14000 Da. At the same time, add 2 mL of PBS (pH=7.4) to the dialysis bag. After sealing the dialysis bag, place it in a stoppered conical flask containing 20 mL of PBS (pH=7.4) as the release medium, ensuring that the solid portion of the dialysis bag is completely immersed in the buffer solution.

[0119] S3. Place all three samples in a constant-temperature shaker at 100 r / min and shake at 37°C in the dark. At different time points (0–48 h), take 2 mL of solution from each conical flask and add an equal volume of PBS (pH 7.4). Three control groups were set up for each type of sample. Number the solutions taken at different time points, filter them through a 0.45 μm microporous membrane, and store them sealed at room temperature in the dark.

[0120] S4. After diluting the solutions taken at different time points, the absorbance was measured using a UV spectrophotometer at a wavelength of λmax = 260 nm. The concentration of S-(-)-Nicotine was calculated based on the UV standard curve of S-(-)-Nicotine. The concentration was multiplied by the corresponding dilution factor to obtain the reagent concentration, and then the cumulative release of S-(-)-Nicotine was calculated. M t ) and cumulative release rate ( Q t ).

[0121] M t ; Q t %= × 100%

[0122] V is the volume of each sample taken; c n-1 The mass concentration of S-(-)-nicotine in the solution sampled in the (n-1)th time; V0 is the total volume of the release medium; c n M represents the mass concentration of S-(-)-nicotine in the solution sampled in the nth time; loading Drug-loaded hydrogels or Nicorettes ® The mass of S-(-)-nicotine loaded in a 2cm×2cm area.

[0123] The experimental results and conclusions are as follows:

[0124] The cumulative release rates of the three samples are shown in Table 3. Simultaneously, the Higuchi equation model (tortuous pore diffusion) was selected to fit the in vitro cumulative release patterns of the three patch systems, thereby determining the drug release effect. The cumulative release curves are attached. Figure 6 See Appendix for the in vitro release equation of Higuchi. Figure 7 .in, Figure 7 A is the Higuchi equation fitting curve for the in vitro release of S-(-)-Nicotine-NFL; Figure 7 B is the Higuchi equation fitting curve for the in vitro release of S-(-)-Nicotine-HGP; Figure 7C is the Higuchi equation fitting curve for the in vitro release of S-(-)-Nicotine-NFL-HGP; Figure 7 D is Nicorette ® The in vitro release Higuchi equation fitting curve is shown. The relevant data, results, and conclusions are as follows:

[0125] Table 3S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP and Nicorette ®

[0126] The cumulative in vitro drug release rate (n=3)

[0127]

[0128] The cumulative release rate of S-(-)-Nicotine-HGP over 48 hours was 68.7%, with the majority of release occurring in the first 12 hours (57.1%). The cumulative release rate in the remaining 12 hours was approximately 11%, indicating that the release was most effective only in the first 12 hours. The cumulative release rate of S-(-)-Nicotine-NFL-HGP over 48 hours was 64.8%, with the release rate showing a uniform upward trend throughout the 48 hours, indicating good release throughout the period. Compared to S-(-)-Nicotine-HGP, the cumulative release rates of the two are not significantly different. ® The cumulative drug release rate of the patch after 48 hours was 59.2%, and the cumulative drug release rate in the first 8 hours had reached 50.1%. However, the cumulative release rate in the following 40 hours was only 9.1%, indicating that the drug release effect was better only in the first 8 hours.

[0129] The Higuchi equation for drug release from S-(-)-Nicotine-NFL is y = 11.69x. 1 / 2 +7.19; R 2 =0.92529, indicating a high degree of fit (R²). 2 >0.9), the drug release effect within 48 h was good, but liposomes were not used as the final drug delivery system in this study. Due to their superior sustained-release effect, they were combined with a hydrogel system to prepare the final patch system; the Higuchi equation for the drug release of S-(-)-Nicotine-HGP is y=10.19x 1 / 2 +8.45; R 2 =0.85781, indicating a low goodness of fit (R²). 2 <0.9), poor drug release effect within 48 h; the Higuchi equation for the drug release of S-(-)-Nicotine-NFL-HGP is y=9.47x1 / 2 -1.13; R 2 =0.99412, indicating a high good fit, suggesting that the S-(-)-Nicotine-NFL-HGP system exhibits good, stable, and long-lasting sustained-release kinetics within 48 h; the Higuchi equation for drug release from commercially available S-(-)-Nicotine patches is y=9.36x. 1 / 2 +6.49; R 2 =0.82672, indicating a low goodness of fit (R²). 2 >0.9), poor drug release effect within 48 hours.

[0130] Example 11: Transdermal test of S-(-)-Nicotine-NFL-HGP

[0131] The research methodology is as follows:

[0132] S1. After feeding healthy mice for 24 hours, shave off their fur, euthanize them by disembowelment, and take the abdominal skin. Completely remove the subcutaneous fat, clean the skin with physiological saline, absorb the surface moisture with filter paper, cut the mouse skin into appropriate sizes, and fix it on the Franz vertical diffusion cell with the stratum corneum facing upward.

[0133] S2. Cut the prepared S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, and Nicorette® into patches of the same size and suitable dimensions (2.54 cm). 2 Then, the patch is attached to the surface of the mouse skin, and air bubbles between the patch and the skin are removed. It is then fixed between the supply chamber and the receiving chamber of the vertical diffusion cell with a spring clip. A magnetic stir bar is placed in the receiving chamber, and 7.5 mL of PBS solution with pH=7.4 is poured in as the receiving solution. Air bubbles are removed, and the receiving cell is placed in a 37°C constant temperature magnetic stirrer with the speed set to 800 rpm.

[0134] S3. Take 2 mL of sample from the receiving cell at 0~48 h and add an equal volume of PBS solution with pH=7.4. Filter the sample solution through a 0.45μm microporous membrane, number the solutions taken at different time points, and store them in a sealed container at room temperature away from light.

[0135] S4. After diluting the solutions taken at different time points, the absorbance was measured using a UV spectrophotometer at a wavelength of λmax = 260 nm. The concentration of S-(-)-Nicotine was calculated based on the S-(-)-Nicotine UV standard curve. The concentration was multiplied by the corresponding dilution factor to obtain the concentration of the released sample, and then the cumulative transmittance of S-(-)-Nicotine per unit area was calculated. Q nThree parallel tests were conducted for each of the three samples, and the results were averaged.

[0136] Q n ) / A

[0137] V is the volume of each sample taken; c n-1 The mass concentration of S-(-)-nicotine in the solution sampled in the (n-1)th time; V0 is the total volume of the release medium; c n denoted as S-(-)-nicotine in the solution sampled for the nth time; A is the effective contact area of ​​the diffusion cell.

[0138] S5. After the transdermal test, the patch was slowly removed, and the isolated rat skin was washed with physiological saline to ensure no patch residue remained. After drying the skin, it was ground into a paste using a mortar and pestle, dissolved in an appropriate amount of ethanol, and sonicated for 30 min. The paste was then centrifuged at 3500 r / min for 20 min, and the supernatant was collected. The paste was then passed through a 0.22 μm organic membrane, and the S-(-)-Nicotine content was determined using the UV standard curve method. The amount of dermal skin retained per unit area of ​​rat skin was calculated. Each sample group was tested three times, and the average value was taken.

[0139] The experimental results and conclusions are as follows:

[0140] The cumulative permeation curves of the three samples obtained from the transdermal test are attached. Figure 8 , Among them, S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, Nicorette ® The Higuchi equation model fitting curve for cumulative transdermal drug release per unit area over 48 hours is attached. Figure 9 ,in, Figure 9 A is the Higuchi equation fitting curve of cumulative transdermal drug release per unit area of ​​S-(-)-Nicotine-HGP over 48 h; Figure 9 B is the Higuchi equation fitting curve of cumulative transdermal drug release per unit area of ​​S-(-)-Nicotine-NFL-HGP over 48 h; Figure 9 C is the Higuchi equation fitting curve of cumulative transdermal drug release per unit area of ​​Nicorette® over 48 h; Figure 10 For S-(-)-Nicotine-HGP, S-(-)-Nicotine-NFL-HGP, Nicorette ® A comparison chart of dermal retention results. Specific experimental data are as follows:

[0141] Table 4. Cumulative drug permeation per unit area of ​​three samples over 48 h (n=3)

[0142]

[0143] The cumulative drug permeation per unit area of ​​the S-(-)-Nicotine-HGP patch over 48 hours was (1164±35.5) μg / cm². 2 The concentration was the highest among the three samples; the dermal retention was (11.4±2.8) μg / cm³. 2 The value was the lowest among the three samples. The Higuchi equation fitting curve was: y = 170.26x 1 / 2 +129.08, R 2 =0.91043; the equation has a low goodness of fit, poor sustained-release effect, and basically does not meet the transdermal drug release rules of sustained-release formulations. The cumulative drug permeation per unit area of ​​S-(-)-Nicotine-NFL-HGP patch over 48 hours was (1047±25.1) μg / cm². 2 The concentration was lower than that of S-(-)-Nicotine-HP patch, but the difference was not significant; the dermal retention was (35.1±4.2) μg / cm³. 2 The concentration was the highest among the three samples, indicating that the drug carrier accumulated in the rat dermis, forming a good "drug reservoir" effect and slowly releasing the drug. The Higuchi equation fitting curve was: y = 162.07x 1 / 2 -47.21, R 2 =0.99192; the equation has a high good fit and meets the transdermal drug release pattern of sustained-release formulations. Combined with the release curve, it can be determined that it has a long-acting drug release effect. The cumulative drug permeation per unit area of ​​Nicorette® patch over 48 hours is (832±23.8) μg / cm². 2 The lowest among the three samples; the dermal retention was (20.9±5.7) μg / cm³. 2 It is slightly higher than that of S-(-)-Nicotine-HGP patches. The Higuchi equation fitting curve is: y=108.31x 1 / 2 +139.77, R 2 =0.93616; the equation fit is also lower than that of S-(-)-Nicotine-NFL-HGP patch, and the slow and long-lasting drug release effect is not as good as that of S-(-)-Nicotine-NFL-HGP.

[0144] In summary, the cumulative transdermal drug delivery per unit area of ​​S-(-)-Nicotine-NFL-HGP over 48 hours meets the transdermal drug release characteristics of long-acting sustained-release and controlled-release formulations, exhibiting the best performance among the three patches. It essentially meets the drug requirements of smoking cessation patients within 48 hours of using the S-(-)-Nicotine-NFL-HGP patch. Therefore, the sustained-release effect of the S-(-)-Nicotine-NFL-HGP patch can prolong the dosing cycle, reduce the dosing frequency, meet the body's need for S-(-)-Nicotine for longer periods, increase patient compliance, and improve therapeutic efficacy.

[0145] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention, all of which should be included within the protection scope of the present invention.

Claims

1. A method for preparing an S-(-)-nicotine nano-flexible liposome hydrogel patch, characterized in that, Includes the following steps: 1) Nanoflexible liposomes loaded with S-(-)-nicotine were prepared by thin-film dispersion method; 2) Mix the S-(-)-nicotine-loaded flexible liposomes obtained in step 1) with an aqueous matrix to obtain a mixture; 3) Add the mixture obtained in step 2) to the hydrogel matrix to obtain the drug-loaded matrix; 4) The drug-loaded matrix obtained in step 3) is uniformly coated on the substrate, dried and cured, and covered with a protective layer to obtain the S-(-)-nicotinic acid nano-flexible liposome hydrogel patch. In step 1), the S-(-)-nicotinic acid-loaded nano-flexible liposomes use phospholipids and cholesterol as phospholipid bilayer framework materials; the membrane softener is 1,2-propanediol.

2. The preparation method of the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 1, characterized in that, The phospholipid is selected from at least one of phosphatidylethanolamine, phosphatidylcholine, and hydrogenated lecithin.

3. The preparation method of the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 2, characterized in that, The phosphatidylcholine is L-α-phosphatidylcholine.

4. The preparation method of the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 3, characterized in that, The S-(-)-nicotinic acid-loaded nano-flexible liposomes use L-α-phosphatidylcholine and cholesterol as phospholipid bilayer framework materials, with the mass ratio of L-α-phosphatidylcholine to cholesterol being (34~38):1; and the mass ratio of 1,2-propanediol to L-α-phosphatidylcholine being (8~12):

1.

5. The preparation method of the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 4, characterized in that, Step 1) further includes the following steps: S1. Place L-α-phosphatidylcholine and cholesterol in a reaction vessel, add solvent, sonicate, homogenize, and dry under reduced pressure until a lipid film forms on the wall of the reaction vessel. S2. Dissolve S-(-)-nicotine in a first 1,2-propanediol aqueous solution, hydrate it with the lipid film obtained in step S1 under vacuum, add a second 1,2-propanediol aqueous solution, mix and homogenize to obtain S-(-)-nicotine-loaded flexible liposomes.

6. The preparation method of the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 5, characterized in that, In step S1, the solvent is selected from C1-C6 alcohols.

7. The preparation method of the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 5, characterized in that, In step S2, the hydration temperature is 25-45℃, the hydration time is 1-3 hours, the first 1,2-propanediol aqueous solution and the second 1,2-propanediol aqueous solution have the same concentration and volume, and the concentration of both the first and second 1,2-propanediol aqueous solutions is 1.5-2 g / mL. The volume ratio of S-(-)-nicotine to the first 1,2-propanediol aqueous solution is (2.5-3.5):1×10 3 .

8. The preparation method of the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 1, characterized in that, In step 2), the aqueous matrix includes 0.16-0.2 parts by weight of tartaric acid, 0.08-0.1 parts by weight of polyvinylpyrrolidone, and 4.8-5.2 parts by weight of water. The mass-to-volume ratio of the aqueous matrix to the S-(-)-nicotine nano-flexible liposomes is 0.25-0.28 g / mL.

9. The method for preparing the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 1, characterized in that, In step 3), the hydrogel matrix is ​​obtained by mixing and stirring sodium polyacrylate, aluminum glycinate, disodium ethylenediaminetetraacetate, azone, glycerol, and 1,2-propanediol. The hydrogel matrix includes 0.9-1 parts by weight of sodium polyacrylate, 0.12-0.18 parts by weight of aluminum glycinate, 0.12-0.13 parts by weight of disodium ethylenediaminetetraacetate, 0.08-0.1 parts by weight of azone, 3.8-4.2 parts by weight of glycerol, and 1.8-2.2 parts by weight of 1,2-propanediol. The mass-to-volume ratio of the hydrogel matrix to the mixture is 0.28-0.32 g / mL.

10. The method for preparing the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 1, characterized in that, In step 3), under the first stirring, the mixture obtained in step 2) is added to the hydrogel matrix, and then stirred again to obtain the drug-loaded matrix. The first stirring is carried out at 50~80℃ and the stirring speed is 300~700r / min. The stirring time of the second stirring is 10~60 min. The mixture obtained in step 2) is added dropwise to the hydrogel matrix, and the dropping rate is controlled to be 10~30% / min of the total weight of the mixture.

11. The method for preparing the S-(-)-nicotine nano-flexible liposome hydrogel patch according to claim 1, characterized in that, In step 4), the drying and curing temperature is controlled at 40~60℃, and the drying and curing time is controlled at 2~12 h.

Images