An acetyl gastrodin transdermal patch and a preparation method thereof

Abstract

The application belongs to the technical field of pharmaceutical preparations, and specifically discloses an acetyl gastrodin transdermal patch and a preparation method thereof. The transdermal patch takes acetyl gastrodin as an effective component, is encapsulated in cubic phase liposomes composed of glycerol monostearate, cholesterol and Pluronic F127, is dispersed in a complex matrix of hydroxypropyl methyl cellulose and ethyl cellulose, and is matched with a complex transdermal enhancer of glyceryl caprylate / caprate and ethanol. In the preparation, the liposome material is first melted to load the drug, and then drug-loaded cubic phase liposomes are formed through ice water bath ultrasonic treatment. Then, the drug-loaded cubic phase liposomes are mixed with the polymer matrix and excipients, are defoamed, are coated, and are dried in two steps to be shaped. The cubic phase liposomes are used to significantly improve the drug penetration rate and stability, realize 24-hour smooth drug release, efficiently penetrate the skin and the blood-brain barrier, significantly inhibit non-small cell lung cancer brain metastases, improve tumor secondary anxiety and depression, have excellent skin compatibility, and provide a new non-invasive synergistic treatment scheme for brain tumors and concurrent emotional disorders.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical formulation technology, specifically relating to an acetyl gastrodin transdermal patch for inhibiting brain tumors and improving anxiety and depression, and its preparation method. Background Technology

[0002] According to global cancer statistics in 2020, lung cancer ranks first in both incidence and mortality among malignant tumors. Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancers, and late-stage brain metastasis is a major cause of high mortality. Clinical data shows that approximately 15%–20% of NSCLC patients already have brain metastases at initial diagnosis, and the incidence of brain metastases throughout the disease course is as high as 40%. Current clinical treatment for lung cancer brain metastases mainly employs single or combined regimens such as surgery, stereotactic radiotherapy, and molecular targeted therapy. However, the unique physiological barriers of the brain and the complexity of the tumor microenvironment make it a major challenge in clinical treatment.

[0003] In the theoretical system of Traditional Chinese Medicine (TCM), lung cancer brain metastases are often categorized under "headache" and "dizziness," with the core pathogenesis being internal liver wind and liver stagnation obstructing the collaterals. The main treatment principle is to calm the liver, extinguish wind, and dispel wind to unblock the collaterals. Gastrodia elata, a traditional and precious Chinese herb, possesses the effects of calming wind and stopping spasms, suppressing liver yang, and dispelling wind to unblock the collaterals. Clinically, it is often used in combination with other herbs for the treatment of dizziness, headaches, and brain tumors. Gastrodin is the core active ingredient of Gastrodia elata, which can cross the blood-brain barrier to exert neuroprotective effects. It has definite efficacy against central nervous system diseases such as anxiety, depression, cerebral ischemia-reperfusion injury, and cerebral infarction. It also shows significant potential in inhibiting neuroinflammation, regulating the tumor brain metastasis microenvironment, and antidepressant effects. However, the strong hydrophilicity of natural gastrodin makes drug development and delivery difficult. Although acetylgastrodin obtained through structural modification has improved lipophilicity, it still faces significant clinical and formulation bottlenecks: low bioavailability and significant first-pass effect when administered orally; and problems such as burst release, insufficient long-acting transdermal rate, and phase separation when directly dispersed in conventional gel matrices. For brain tumor patients with anxiety and depression who require long-term maintenance of effective drug concentrations in the brain, existing drug delivery systems have poor compliance and cannot meet the clinical needs for long-acting, stable, and non-invasive drug delivery.

[0004] Cubosomes are highly ordered liquid crystal nanocarriers formed by the self-assembly of amphiphilic lipids. They possess a unique dual-continuous aqueous channel and lipid bilayer structure, which can significantly improve the encapsulation efficiency of moderately lipophilic drugs and substantially reduce the barrier resistance of the stratum corneum. Therefore, constructing an acetylgastrodin transdermal patch integrating cubic liposome nanodelivery technology and a polymeric sustained-release framework has significant scientific and clinical value for achieving efficient transdermal and blood-brain barrier drug delivery, as well as non-invasive synergistic treatment of brain tumors and their secondary mood disorders. Summary of the Invention

[0005] The purpose of this invention is to address the problems of low transdermal efficiency, uncontrollable release, and poor stability of conventional acetyl gastrodin matrices. This invention provides an acetyl gastrodin transdermal patch and its preparation method, using cubic liposomes as a carrier and composite polymers as a framework to achieve stable transdermal drug release over 24 hours, efficiently penetrate the blood-brain barrier, inhibit brain metastases from non-small cell lung cancer, improve secondary anxiety and depression, and enhance patient medication adherence and clinical treatment efficacy.

[0006] To achieve the above objectives, one of the technical solutions adopted by the present invention is to provide an acetyl gastrodin transdermal patch, comprising a backing layer, a drug-containing matrix layer, and an anti-adhesive layer. The active ingredient of the drug-containing matrix layer is acetyl gastrodin, and the acetyl gastrodin is encapsulated in cubic liposomes. The cubic liposomes are composed of glyceryl monostearate, cholesterol, and the surfactant Pluronic F127, which can effectively encapsulate the acetyl gastrodin and significantly improve the skin permeability of the drug by utilizing its high-curvature aqueous channels. The drug-containing matrix layer also includes a hydrophilic polymer backbone, a hydrophobic polymer backbone, a transdermal penetration enhancer, and medical excipients.

[0007] Furthermore, the mass ratio of the glyceryl monostearate, cholesterol, and surfactant Pluronic F127 is 10:2:1.

[0008] Furthermore, the hydrophilic polymer backbone is hydroxypropyl methylcellulose (HPMC) to provide a soft gel matrix; the hydrophobic polymer backbone is ethyl cellulose (EC) to construct a hydrophobic network and control the drug release rate; the transdermal penetration enhancer is a combination of caprylic / capric glyceride and anhydrous ethanol, which work synergistically with cubic liposomes to reduce stratum corneum resistance.

[0009] Furthermore, the hydroxypropyl methylcellulose (HPMC) is selected from high viscosity grade, specifically selected from at least one of K100M, K4M, and E4M, with K100M being preferred.

[0010] Furthermore, the medical excipients include a pressure-sensitive adhesive, a plasticizer, and a preservative; the pressure-sensitive adhesive is a polyacrylate pressure-sensitive adhesive (PSA); the plasticizer is glycerin; and the preservative is phenoxyethanol.

[0011] Further, by mass percentage, the drug-containing matrix layer comprises the following raw materials: acetyl gastrodin 1%–10%, cubic phase liposomes 5%–20%, polymer backbone 20%–50%, transdermal penetration enhancer 2%–15%, pressure-sensitive adhesive 5%–20%, plasticizer 1%–5%, preservative 0.05%–0.5%, with the balance being medical volatile solvents.

[0012] To achieve the above objectives, the second technical solution adopted by the present invention is to provide a method for preparing the above-mentioned acetyl gastrodin transdermal patch, comprising the following steps:

[0013] (1) Preparation of drug-loaded cubic liposome premix: Glyceryl monostearate, cholesterol and surfactant Pluronic F127 were heated to melt, cooled to room temperature to form cubic liposome gel, acetyl gastrodin was added, and ultrasonically dispersed under ice bath to obtain drug-loaded cubic liposome premix.

[0014] (2) Preparation of polymer matrix: The hydrophilic polymer backbone and the hydrophobic polymer backbone are dissolved separately and then mixed to form a polymer matrix;

[0015] (3) Preparation of drug-containing slurry: Add the drug-loaded cubic liposome premix from step (1) to the polymer matrix from step (2), add the transdermal penetration enhancer and medical excipients in sequence, stir evenly and degas under vacuum to obtain the drug-containing slurry;

[0016] (4) Coating and drying: The drug-containing slurry is evenly coated on the backing layer, and after drying, the solvent evaporates to form a film to form a drug-containing matrix layer. Finally, an anti-adhesive layer is laminated and cut into appropriate sizes, and then sealed and packaged to obtain the acetyl gastrodin transdermal patch.

[0017] Further, the drying in step (4) is as follows: first, let it stand at room temperature for 24 hours to dry, and then transfer it to a vacuum drying oven at 35-45°C for 10-14 hours to dry.

[0018] The present invention also provides the application of the above-mentioned acetyl gastrodin transdermal patch in the preparation of a drug for treating brain tumors or mood disorders secondary to the deterioration of the brain tumor microenvironment; wherein the brain tumor is a non-small cell lung cancer brain metastasis, and the mood disorder is anxiety or depression; the acetyl gastrodin transdermal patch can efficiently penetrate the blood-brain barrier and exert a synergistic therapeutic effect by regulating the tumor microenvironment in the brain and anti-neuroinflammatory.

[0019] Compared with the prior art, the present invention has the following outstanding advantages:

[0020] This invention innovatively employs cubic liposome technology to encapsulate acetyl gastrodin, which significantly overcomes the skin's physical barrier to transdermal drug absorption and substantially increases drug penetration compared to traditional gel matrices. By constructing a hydrophilic-hydrophobic composite polymer framework using hydroxypropyl methylcellulose and ethylcellulose, it effectively inhibits drug burst release caused by liposome rupture, achieving a stable and controllable release of acetyl gastrodin for up to 24 hours, significantly improving long-term drug compliance in patients with brain tumors and anxiety / depression. The resulting patch exhibits low skin irritation and excellent biocompatibility, making it safe for long-term non-invasive application. This invention overcomes the limitations of traditional single-target therapy, enabling the drug to efficiently penetrate the blood-brain barrier, and possesses a dual therapeutic effect of inhibiting the colonization and growth of non-small cell lung cancer brain metastases and improving tumor-related anxiety and depressive disorders. It provides a novel non-invasive synergistic drug delivery strategy for brain tumors and their associated neuropsychiatric symptoms, possessing extremely high clinical translational and application value. Attached Figure Description

[0021] Figure 1 These are typical intracranial bioluminescence imaging images and box plots of intracranial bioluminescence intensity (BLI) for each group of mice in test example 4. P<0.001).

[0022] Figure 2 This is a dynamic curve showing the change of intracranial tumor fluorescence signal values ​​over time in each group of mice in test example 4.

[0023] Figure 3 This is a bar chart comparing the fluorescence signal values ​​of intracranial tumors in each group of mice on day 21 in test example 4.

[0024] Figure 4 These are the behavioral results of the open field (OFT) experiments performed on each group of mice in Test Example 4.

[0025] Figure 5 This is a bar chart showing the quantitative results of immobility time in the tail suspension test (TST) for each group of mice in Test Example 4. Detailed Implementation

[0026] The technical solution of the present invention will be further described below with reference to the embodiments. However, the scope of protection of the present invention includes, but is not limited to, these embodiments. Any changes or equivalent substitutions that do not depart from the concept of the present invention are included within the scope of protection of the present invention.

[0027] The instruments and reagents used in the embodiments of this invention are all commercially available products. The model and manufacturer information of some reagents are as follows:

[0028] All reagents used in the embodiments of this invention are commercially available medical or pharmaceutical grade.

[0029] Example 1

[0030] An acetyl gastrodin transdermal patch, the drug-containing matrix layer of which comprises the following raw materials: acetyl gastrodin 5%, glyceryl monostearate 10%, cholesterol 2%, Pluronic F127 1%, hydroxypropyl methylcellulose (HPMC K100M) 25%, ethyl cellulose (EC) 15%, caprylic / capric glyceride 5%, anhydrous ethanol 5%, polyacrylate pressure-sensitive adhesive (PSA) 12%, glycerin 3%, phenoxyethanol 0.1%, and ethanol / purified water mixed solvent (volume ratio 3:7) 16.9%.

[0031] Its preparation method includes the following steps:

[0032] (1) Preparation of drug-loaded cubic liposome premix: Glyceryl monostearate, cholesterol and surfactant Pluronic F127 were melted in a 70°C constant temperature water bath and then cooled to room temperature to form a transparent cubic liposome gel; Acetyl gastrodin was accurately weighed and added to it, and the mixture was ultrasonically treated with a probe under ice bath for 10 minutes to obtain the drug-loaded cubic liposome premix.

[0033] (2) Preparation of polymer matrix: Dissolve HPMC K100M and EC in a mixed solvent of ethanol / purified water, and then mix them evenly to form a polymer matrix;

[0034] (3) Preparation of drug-containing slurry: The drug-loaded cubic liposome premix from step (1) is slowly added to the polymer matrix from step (2), and octanoic acid / capric acid glyceride, glycerol, phenoxyethanol, ethanol and PSA are added in sequence. After each addition, the mixture is stirred vigorously for 10 to 15 minutes until the system is homogeneous. Vacuum degassing is then performed to obtain the drug-containing slurry.

[0035] (4) Coating and drying: The drug-containing slurry is evenly coated on the polyester / aluminum foil composite substrate (backing layer), and allowed to stand and dry at room temperature for 24 h. Then it is transferred to a vacuum drying oven and vacuum dried at 40°C for 12 hours to completely remove residual solvent, so that the drug-containing slurry forms a film to form a drug-containing matrix layer. Finally, an anti-adhesive layer is laminated and cut into a suitable size (10 cm × 5 cm). The acetyl gastrodin transdermal patch is then sealed and packaged.

[0036] Examples 2-4

[0037] Table 1. Composition of the drug-containing matrix layer of acetyl gastrodin transdermal patches in Examples 2-4

[0038]

[0039] Based on Example 1, the effect of the amount of transdermal penetration enhancer (caprylic acid / capric acid glyceride) was investigated. The raw material composition of the drug-containing matrix layer is shown in Table 1, and the preparation method is the same as in Example 1.

[0040] Comparative Example 1 (liposomes without cubic phase)

[0041] Based on Example 1, glyceryl monostearate, cholesterol, and Pluronic F127 were removed, and acetyl gastrodin was directly dispersed in the polymer matrix. The rest were the same as in Example 1, except that an equal amount of solvent was used instead.

[0042] Comparative Example 2 (without transdermal penetration enhancer)

[0043] Based on Example 1, caprylic / capric glyceride and anhydrous ethanol used for penetration enhancement were removed and replaced with an equal amount of solvent, while the rest remained the same as in Example 1.

[0044] Test Example 1: Adhesion Strength Measurement

[0045] According to the Adhesion Test Method in General Chapter 0952 of Part IV of the 2020 edition of the Chinese Pharmacopoeia, the initial tack (slant rolling ball method), holding power and peel strength (180° peel method) of the acetyl gastrodin transdermal patches of Examples 1-4 and Comparative Examples 1-2 were determined, and the results are shown in Table 2.

[0046] Table 2. Adhesion test results of acetyl gastrodin transdermal patches in Examples 1-4 and Comparative Examples 1-2.

[0047]

[0048] The test results showed that octanoic acid / capric acid glyceride, as a polar lipid, not only enhanced penetration but also significantly softened and plasticized the polymer backbone. When its dosage increased to 8% (Example 3) and 10% (Example 4), the patch matrix network became excessively loose. Although the initial tack increased, the holding power and peel strength both decreased sharply, easily leading to patch cohesive failure and cold flow of the adhesive layer. Example 1 (5% octanoic acid / capric acid glyceride) maintained excellent initial tack (15#) while the holding power and peel strength were within the ideal application range, resulting in the best overall adhesion performance.

[0049] Test Example 2: Evaluation of Skin Irritation and Allergy

[0050] To evaluate the skin safety of the acetyl gastrodin transdermal patches of Examples 1-4 and Comparative Examples 1-2, their skin irritation and sensitization were investigated.

[0051] Experimental Methods: Healthy adult rabbits were used. The acetyl gastrodin transdermal patches from Examples 1-4 and Comparative Examples 1-2 were cut into appropriate sizes and applied to the skin on the back of the rabbits for 24 hours. After removing the patches, the skin residue was cleaned with warm water. Skin erythema and edema reactions were observed and recorded at 1 hour, 24 hours, 48 ​​hours, and 72 hours after removal. The primary irritation index (PII) was calculated according to the skin irritation scoring criteria shown in Table 3.

[0052] Primary Stimulation Index (PII) = (Total erythema score at each observation time point + Total edema score) ÷ (Number of observations × Number of animals).

[0053] Table 3. Skin Irritation Reaction Scoring Criteria

[0054]

[0055] Irritation determination: PII 0-0.49 is non-irritating; 0.5-2.99 is mildly irritating; 3.0-5.99 is moderately irritating; 6.0-8.0 is severely irritating. The skin irritation test results of each group of patches are shown in Table 4.

[0056] Table 4. Primary Skin Irritation Index (PII) of Acetyl Gastrodin Transdermal Patches

[0057]

[0058] Results analysis: The primary irritation index (PII) of all groups of patches was less than 0.49, meeting the criteria for non-irritation. Even in Example 4, where the penetration enhancer dosage was as high as 10%, no obvious allergic reactions such as erythema or edema occurred. The results indicate that the present invention, using cubic liposomes to encapsulate acetyl gastrodin and selecting mild caprylic / capric glyceride as a transdermal permeability enhancer, effectively avoids the epidermal irritation easily caused by traditional volatile oils and short-chain fatty acids. The patch exhibits excellent skin biocompatibility and is suitable for long-term use by patients with central nervous system diseases.

[0059] Test Example 3: In vitro transdermal test

[0060] Experimental objective: To investigate the in vitro transdermal permeation kinetics of acetyl gastrodin in Example 1 (5% caprylic / capric glyceride), Comparative Example 1 (without cubic liposomes), and Comparative Example 2 (without transdermal permeation enhancer), and to verify the synergistic effect of cubic liposomes and the composite permeation-enhancing system.

[0061] Experimental method: In vitro transdermal experiments were conducted using a modified Franz diffusion cell, with an effective transdermal area A of 3.14 cm². 2The receiving pool volume V is 15 mL. Skin from the back of a healthy hairless mouse was taken, subcutaneous fat was removed, and the skin was washed with physiological saline. This skin was then fixed between the supply and receiving pools, ensuring the stratum corneum faces the supply pool and the dermis faces the receiving pool. The acetylgastrodin transdermal patches from Examples 1, 1, and 2 were accurately cut to 3.14 cm pieces. 2 The circular patch is applied tightly to the stratum corneum of the skin. The receiving solution is PBS buffer at pH 7.4, the water bath temperature is controlled at 32±0.5℃ (simulating the surface temperature of human skin), and the magnetic stirring speed is kept constant at 300 rpm.

[0062] Sampling and determination: At 2 h, 4 h, 8 h, 12 h and 24 h after the start of the experiment, 1.0 mL of the receiving solution was taken from the receiving cell as the test solution, and an equal volume of fresh receiving solution at the same temperature was immediately added. The concentration of acetylgastrodin in the receiving solution at each time point was determined by high performance liquid chromatography (HPLC).

[0063] Data processing: Calculate the cumulative transdermal dose Qn (μg / cm²) per unit area using the following formula. 2 ):

[0064]

[0065] In the formula: is the concentration of acetyl gastrodin in the receiving solution at the nth sampling time (μg / mL); V is the total volume of the receiving cell (mL). For the i-th time (i n) Drug concentration at the time of sampling (μg / mL); The volume of fresh receiving fluid to be replenished (mL, consistent with the sampling volume); A is the effective transdermal area (cm²). 2 ).

[0066] The experimental results are shown in Table 5: The 24-hour cumulative transdermal absorption and steady-state transdermal rate of Example 1 were significantly higher than those of Comparative Example 1 and Comparative Example 2, and the transdermal lag time was significantly shortened. The results indicate that this invention significantly improves the transdermal penetration efficiency of acetyl gastrodin through the dual synergistic effect of cubic liposomes and the caprylic / capric glyceride + ethanol composite permeation-enhancing system; the unique high-curvature aqueous channels and membrane fusion effect of cubic liposomes can effectively break through the skin's stratum corneum barrier, providing a solid pharmaceutical basis for the drug's in vivo penetration of the blood-brain barrier and inhibition of NSCLC brain metastases.

[0067] Table 5 Comparison of in vitro transdermal parameters

[0068]

[0069] Test Example 4: In vivo efficacy evaluation of acetyl gastrodin transdermal patch against brain metastases from non-small cell lung cancer

[0070] 1. Experimental objective: To investigate the inhibitory effect of the acetyl gastrodin transdermal patch of Example 1 on brain metastasis and colonization in non-small cell lung cancer (NSCLC) in vivo, and its effect on improving anxiety and depressive mood disorders secondary to tumor microenvironment deterioration.

[0071] 2. Instruments and Materials

[0072] Experimental instruments: Incubator (Thermo Fisher Scientific (China) Co., Ltd.); Clean bench (Jinan Gansi Biotechnology Co., Ltd.); Inverted microscope (Leica GmbH, Germany); Centrifuge (SCILOGEX, USA); Stereoscopic system; General-purpose small animal anesthetic (Reward Life Technology Co., Ltd.); FX-PRO small animal in vivo imaging system (BRUKER, USA).

[0073] Experimental materials: PC-9-Luc cells (purchased from Guangzhou Saiku Biotechnology Co., Ltd., and cryopreserved in the Biochemistry and Pharmacology Laboratory of Bengbu Medical University); DMEM basal medium (Gibco); fetal bovine serum (ExCell Bio); trypsin cell digestion solution (NCM); penicillin-streptomycin (Biosharp); sodium carboxymethyl cellulose (Sigma); D-fluorescein potassium salt (APExBIO); positive control drug aumolertinib (administered by gavage); acetylgastrodin transdermal patch and corresponding blank matrix patch prepared in Example 1 of this invention.

[0074] Experimental animals: SPF-grade female BALB / c nude mice, purchased from Hangzhou Ziyuan Experimental Animal Technology Co., Ltd.

[0075] 3. Establishment of an animal model of brain xenograft in non-small cell lung cancer

[0076] (1) Cell preparation: PC-9-Luc cells were revived and cultured in a clean bench. Cells in the logarithmic growth phase were collected, centrifuged, and resuspended in 1% sodium carboxymethyl cellulose solution to prepare a concentration of 1×10⁻⁶ cells / mL. 7 A cell suspension of 1 cell / mL was stored on ice for later use.

[0077] (2) Positioning and drilling: nude mice were anesthetized and maintained with isoflurane; the nude mice were fixed on a stereotaxic instrument, their heads were disinfected by wiping with alcohol, the skin was cut open to expose the skull, and hydrogen peroxide was applied to remove the surface fascia; the coordinates were zeroed with the anterior fontanelle cross as the origin, and the right brain in situ injection point was located (X-axis right side +1.5 mm, Y-axis posterior -1.0 mm); a skull drill was used to drill a hole at low speed at this point, and skull fragments and meninges were cleaned;

[0078] (3) Cell inoculation: Use a microsyringe to draw 4-5 μL of PC-9-Luc cell suspension, slowly insert the needle into the drill hole to a depth of 3.5 mm below the skull, then withdraw 0.5 mm to form a small reservoir, and slowly inject 3 μL of cell suspension at a rate of 1 μL / min; after injection, leave it in place for 1-2 minutes to prevent backflow, then slowly withdraw the needle, use medical glue to seal the wound, and put the nude mouse back into the constant temperature heating blanket to wait for it to wake up.

[0079] 4. Experimental grouping and drug administration

[0080] Nude mice confirmed by small animal in vivo imaging to have successfully colonized intracranial tumors were selected and randomly divided into three groups: a model control group, a positive drug ametinib group (administered by gavage), and an acetylgastrodin transdermal patch group (Example 1), with 8 mice in each group. The model control group and the positive drug ametinib group received an equal area (6.25 cm²) of transdermal patch at the same site daily. 2 The control group received blank matrix patches (to control for stress variables in animals), with the positive control group additionally administered a specific dose (15 mg / kg) of amitinib solution daily by gavage; the acetyl gastrodin transdermal patch group received the acetyl gastrodin transdermal patch of Example 1 (treatment area 6.25 cm²). 2 Each group was slightly reinforced with medical breathable tape, and the patch was changed and medication was administered daily for 21 consecutive days.

[0081] 5. Small animal in vivo imaging for detecting intracranial tumor growth

[0082] On days 7, 14, and 21 after cell seeding, intracranial tumor growth was dynamically monitored using a small animal in vivo imaging system. Before detection, mice in each group were intraperitoneally injected with D-fluorescein potassium substrate (0.2 mL / mouse). After standing for 5 minutes to allow the substrate to fully react, the mice were anesthetized with isoflurane and placed in the dark chamber of the in vivo imaging system for imaging acquisition. The size of the intracranial tumor burden was quantitatively characterized by the intracranial bioluminescence photon intensity (BLI value). Higher luminescence intensity indicated a larger intracranial tumor volume and more severe malignant progression. The results are as follows: Figure 1 , Figure 2 , Figure 3 As shown. Figure 1 Imaging results showed that: in the model control group, the brains of nude mice exhibited bright red-yellow strong bioluminescent signals, indicating widespread intracranial tumor colonization, continuous malignant proliferation, and an extremely high tumor burden; the brains of the acetyl gastrodin transdermal patch group showed only weak blue-purple fluorescence, and the intracranial BLI signal intensity was significantly lower than that of the model control group. P<0.001); the intracranial fluorescence signal in the positive drug amitinib group also remained at a very low level, and the tumor-suppressing effect was significant. Figure 2Dynamic growth curve results showed that the intracranial tumor in the model control group exhibited explosive and rapid growth, with a BLI value reaching 5.7 × 10⁻⁶ on day 21. 9 In the acetyl gastrodin transdermal patch group, tumor growth was significantly inhibited, and the growth rate was greatly slowed down. On day 21, the intracranial tumor burden was only about 1 / 3 of that in the model control group. In the positive drug amitinib group, the tumor level remained extremely low throughout the process, with no significant progression. Figure 3 Quantitative results at the endpoint showed that the intracranial tumor burden in the model control group was significantly higher than that in the other two groups; the acetyl gastrodin transdermal patch could effectively inhibit the colonization and growth of NSCLC brain metastases in vivo, with a clear tumor-suppressing effect, consistent with the trend of the first-line targeted positive drug amitinib.

[0083] The above in vivo imaging test results fully demonstrate that the acetyl gastrodin transdermal patch of the present invention can efficiently penetrate the blood-brain barrier and significantly delay and inhibit the colonization, proliferation and malignant progression of non-small cell lung cancer brain metastases in vivo, and has excellent in vivo anti-brain metastasis activity.

[0084] 6. Animal behavioral evaluation

[0085] After a 21-day dosing cycle, open field and tail suspension tests were performed on nude mice in each group to systematically evaluate the ameliorative effect of the acetyl gastrodin transdermal patch of this invention on anxiety and depression-like neuropsychiatric abnormalities in NSCLC brain metastases model mice. The experimental results are as follows: Figure 4 , Figure 5 As shown.

[0086] Open Field Test (OFT): Mice were placed in a standardized open field behavior testing chamber. A video trajectory tracking system continuously recorded the mice's spontaneous activity trajectory, total movement distance, number of times they crossed the central thread, and time remaining still for 5 minutes. The mice's anxiety level was assessed by their willingness to explore the central area and their level of spontaneous activity. Higher anxiety levels indicated that mice were more likely to move along the chamber edges and avoid the central open area, manifested as a shorter total movement distance, fewer times they crossed the central thread, and a longer time remaining still. The test results are as follows: Figure 4 As shown: The model control group mice's activity trajectory was highly confined to the edge area of ​​the open field box, with almost no autonomous exploration behavior in the central area, exhibiting typical anxiety-like behavior. Their total movement distance was the shortest, the number of times they crossed the central line was the lowest, and their immobility time was the longest, indicating a significant impairment in autonomous exploration and movement intention. After intervention with the acetyl gastrodin transdermal patch of this invention, the mice's activity trajectory was evenly distributed throughout the open field box, with a significant increase in exploration activity in the central area. The total movement distance and the number of times they crossed the central line were significantly improved compared to the model control group, and the immobility time was significantly shortened. The anxiety-like behavior secondary to tumors was significantly reversed and improved. In contrast, the level of autonomous activity in the positive drug amitinib group and the normal control group increased sequentially, and the willingness to explore the central area increased sequentially. Among them, the activity trajectory of the normal control group mice completely covered the entire open field box, which is the normal autonomous exploration behavior pattern of healthy nude mice.

[0087] Tail Suspension Test (TST): Mice were suspended vertically by their tails using medical tape. The cumulative immobility time in the last 4 minutes of a 6-minute observation period was recorded. Longer immobility time indicated more pronounced despair and abandonment behavior and more severe depressive-like behavior. After each mouse test, the equipment was thoroughly cleaned with 75% ethanol to eliminate cross-interference caused by residual odors. The test results are as follows: Figure 5 As shown: The model control group mice had the longest cumulative immobility time and the most significant despair-like behavior; the acetyl gastrodin transdermal patch group mice had a significantly lower cumulative immobility time than the model control group, and a significantly longer active struggling time, and the depressive-like behavior secondary to tumor brain metastasis was strongly reversed; the positive drug amitinib group had a longer immobility time than the acetyl gastrodin transdermal patch group, and the antidepressant improvement effect was weaker than that of the transdermal patch of this invention.

[0088] The above behavioral test results confirm that NSCLC brain metastases can induce significant anxiety and depression-like neuropsychiatric abnormalities in mice. The acetyl gastrodin transdermal patch of this invention can significantly improve neuroinflammation in the brain tumor microenvironment while inhibiting intracranial tumor progression, and effectively reverse tumor-related anxiety and depression, achieving a dual synergistic therapeutic effect of anti-brain metastases and improvement of neuropsychiatric complications. Moreover, its effect in improving mood disorders is superior to that of the first-line clinical positive targeted drug amitinib, demonstrating excellent clinical translational advantages.

[0089] The embodiments described above are some, but not all, of the embodiments of this application. The detailed description of the embodiments of this application is not intended to limit the scope of the claimed application, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

Claims

1. An acetyl gastrodin transdermal patch, comprising a backing layer, a drug-containing matrix layer, and an anti-adhesive layer, characterized in that, The active ingredient of the drug-containing matrix layer is acetyl gastrodin, which is encapsulated in cubic liposomes. The cubic liposomes are composed of glyceryl monostearate, cholesterol, and the surfactant Pluronic F127. The drug-containing matrix layer also includes a hydrophilic polymer backbone, a hydrophobic polymer backbone, a transdermal penetration enhancer, and medical excipients.

2. The acetylgastrodin transdermal patch according to claim 1, characterized in that: The mass ratio of the glyceryl monostearate, cholesterol, and surfactant Pluronic F127 is 10:2:

1.

3. The acetylgastrodin transdermal patch according to claim 2, characterized in that: The hydrophilic polymer backbone is hydroxypropyl methylcellulose, and the hydrophobic polymer backbone is ethylcellulose; the transdermal penetration enhancer is a combination of caprylic / capric glyceride and anhydrous ethanol.

4. The acetylgastrodin transdermal patch according to claim 3, characterized in that: The hydroxypropyl methylcellulose is selected from at least one of K100M, K4M, and E4M.

5. The acetylgastrodin transdermal patch according to claim 4, characterized in that: The medical excipients include pressure-sensitive adhesives, plasticizers, and preservatives; the pressure-sensitive adhesive is a polyacrylate pressure-sensitive adhesive; and the plasticizer is glycerin.

6. The acetylgastrodin transdermal patch according to claim 5, characterized in that: By mass percentage, the drug-containing matrix layer comprises the following raw materials: acetyl gastrodin 1%–10%, cubic phase liposomes 5%–20%, polymer backbone 20%–50%, transdermal penetration enhancer 2%–15%, pressure-sensitive adhesive 5%–20%, plasticizer 1%–5%, preservative 0.05%–0.5%, with the balance being medical volatile solvents.

7. The method for preparing the acetyl gastrodin transdermal patch according to any one of claims 1 to 6, characterized in that, Includes the following steps: (1) Preparation of drug-loaded cubic liposome premix: Glyceryl monostearate, cholesterol and surfactant Pluronic F127 were heated to melt, cooled to room temperature to form cubic liposome gel, acetyl gastrodin was added, and ultrasonically dispersed under ice bath to obtain drug-loaded cubic liposome premix; (2) Preparation of polymer matrix: The hydrophilic polymer backbone and the hydrophobic polymer backbone are dissolved separately and then mixed to form a polymer matrix; (3) Preparation of drug-containing slurry: Add the drug-loaded cubic liposome premix from step (1) to the polymer matrix from step (2), add the transdermal penetration enhancer and medical excipients in sequence, stir evenly and degas under vacuum to obtain the drug-containing slurry; (4) Coating and drying: The drug-containing slurry is evenly coated on the backing layer, and after drying, the solvent evaporates to form a film to form a drug-containing matrix layer. Finally, an anti-adhesive layer is laminated and cut into appropriate sizes, and then sealed and packaged to obtain the acetyl gastrodin transdermal patch.

8. The preparation method according to claim 7, characterized in that: The drying process in step (4) involves first drying the product at room temperature for 24 hours, and then transferring it to a vacuum drying oven at 35-45°C for 10-14 hours.

9. The use of the acetyl gastrodin transdermal patch according to any one of claims 1 to 6 in the preparation of a medicament for treating brain tumors and / or mood disorders secondary to brain tumors.

10. The application according to claim 9, characterized in that: The brain tumor is a brain metastasis of non-small cell lung cancer, and the mood disorder is anxiety or depression.

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