Natural plant essential oil for calming the mind and promoting sleep, preparation method and sleeping patch

Abstract

The invention relates to the technical field of natural plant essential oils for calming the mind and promoting sleep, and in particular to natural plant essential oils for calming the mind and promoting sleep, a preparation method and a sleeping patch. The natural plant essential oils are prepared from the following components by weight: 20-25 parts of linalool; 15-20 parts of citral; 10-15 parts of alpha-pinene; 8-12 parts of carvacrol; 6-10 parts of (R)-(-)-carvone; 5-9 parts of nerolidol; 4-8 parts of geraniol; 5-10 parts of beta-cyclodextrin; 3-7 parts of soybean lecithin; 1-3 parts of vitamin E; 2-6 parts of spiny jujube kernel oil and 1-4 parts of bergamot oil; linalool and alpha-pinene achieve comprehensive regulation of the sleep-wake cycle, significantly improve the levels of GABA and 5-HT in the brain, shorten the sleep latency and prolong the sleep time; the spiny jujube kernel oil has the effects of sedation, sleep-promoting and anti-anxiety; and the bergamot oil has the effects of sedation, anti-depression and mood improvement.

Description

Technical Field

[0001] The invention relates to the technical field of natural plant essential oils for calming the mind and promoting sleep, and in particular to natural plant essential oils for calming the mind and promoting sleep, a preparation method and a sleeping patch. Background Art

[0002] Sleep disorders are an increasingly prominent health problem in contemporary society, seriously affecting people's quality of life and work efficiency. Long-term sleep problems not only lead to daytime fatigue and cognitive decline, but may also cause or aggravate a variety of chronic diseases such as cardiovascular disease and metabolic disorders.

[0003] At present, the commonly used treatment methods in clinical practice mainly include drug therapy and non-drug therapy. In terms of drug therapy, benzodiazepines (such as diazepam) and non-benzodiazepines (such as zolpidem) are the most commonly prescribed drugs.

[0004] In recent years, plant-derived natural products have received increasing attention in the treatment of sleep disorders. Essential oils such as lavender, valerian and chamomile are popular for their mild hypnotic effects and low side effects. However, existing essential oil preparations still have the following technical problems:

[0005] 1. Single mechanism of action: Most essential oil preparations only target a single target, such as GABA receptors, which makes it difficult to fully regulate the complex sleep-wake cycle.

[0006] 2. Low bioavailability: Essential oil components are volatile, and the efficiency of percutaneous absorption is low, making it difficult to maintain effective blood drug concentrations.

[0007] 3. Poor release control: Existing preparations often show a "high at the beginning and low at the end" release pattern, making it difficult to achieve a sustained and stable therapeutic effect.

[0008] 4. Poor stability: Essential oil components are easily oxidized and photolyzed, and the content of active ingredients decreases rapidly during storage and use.

[0009] 5. Insufficient synergistic effect: Existing formulas often ignore the synergistic effects among multiple components and fail to fully utilize the advantages of the compound.

[0010] 6. Difficulty in individualized dosing: Existing preparations have a single dosage form, making it difficult to adjust the dosage according to the patient's specific circumstances.

[0011] These technical problems have seriously restricted the widespread application of plant essential oils in the treatment of sleep disorders, and there is an urgent need to develop new preparations with multi-target synergy, efficient absorption, long-acting sustained release, good stability and safety. Summary of the invention

[0012] In view of the deficiencies in the prior art, the present invention provides a natural plant essential oil for calming the mind and promoting sleep, a preparation method and a sleeping patch.

[0013] To achieve the above object, the present invention provides the following technical solution: the natural plant essential oil for calming the mind and promoting sleep is prepared from the following components by weight:

[0014] 20-25 parts of linalool; 15-20 parts of citral; 10-15 parts of α-pinene; 8-12 parts of carvacrol; 6-10 parts of (R)-(-)-carvone; 5-9 parts of nerolidol; 4-8 parts of geraniol; 5-10 parts of β-cyclodextrin; 3-7 parts of soybean lecithin; 1-3 parts of vitamin E; 2-6 parts of jujube kernel oil and 1-4 parts of bergamot oil.

[0015] Preferably, the following auxiliary components are also included:

[0016] 1-3 parts of chitosan; 10-15 parts of anhydrous ethanol; 5-8 parts of propylene glycol; 10-15 parts of tricaprin; 40-50 parts of isododecane.

[0017] The preparation method of natural plant essential oil for calming the mind and promoting sleep comprises the following steps:

[0018] (1) preparing an essential oil complex;

[0019] (2) preparing cyclodextrin inclusion complex;

[0020] (3) Preparing the final complex based on the products of steps (1) and (2).

[0021] Preferably, the step (1) comprises:

[0022] (1.1) Mixing linalool, citral, α-pinene, carvacrol, (R)-(-)-carvone, nerolidol, geraniol, jujube kernel oil and bergamot oil at 40-45° C. to obtain a mixed solution A;

[0023] (1.2) Dissolve vitamin E in propylene glycol, add mixed solution A, and stir at 25-30°C for 2 hours to obtain solution B;

[0024] (1.3) Dissolve soybean lecithin in anhydrous ethanol, add it dropwise into solution B at 45-50°C, and stir for 2 hours to form microemulsion C;

[0025] (1.4) The microemulsion C is ultrasonically treated for 15-20 minutes to obtain the essential oil complex D.

[0026] Preferably, the step (2) comprises:

[0027] (2.1) Dissolve β-cyclodextrin in isododecane and stir at 60-65°C until completely dissolved;

[0028] (2.2) Add essential oil complex D to the β-cyclodextrin solution at a rate of 2-3 mL / min and stir for 2 hours;

[0029] (2.3) The mixture was cooled to room temperature, allowed to stand for 12 hours, and then centrifuged to collect the precipitate;

[0030] (2.4) The precipitate was vacuum dried at 40-45°C for 24 hours, ground and sieved to obtain cyclodextrin inclusion complex E.

[0031] Preferably, the step (3) comprises:

[0032] (3.1) Mix the essential oil complex D and the cyclodextrin inclusion complex E and stir at 50-55°C for 1 hour;

[0033] (3.2) Add chitosan pre-dissolved in tricaprin and stir for 30 minutes;

[0034] (3.3) Cool the mixture to 30-35°C, add the remaining isododecane, and stir evenly;

[0035] (3.4) The mixture was homogenized under a vacuum condition of 0.05-0.1 MPa for 10-15 minutes to obtain the final essential oil complex F.

[0036] Preferably, the weight ratio of the essential oil complex D to the cyclodextrin inclusion complex E is 75:8.

[0037] Preferably, the weight ratio of chitosan to tricaprin is 1:3.

[0038] The sleeping patch comprises the natural plant essential oil for calming the mind and promoting sleep.

[0039] The preparation method of the sleeping patch comprises the following steps:

[0040] (1) mixing the essential oil complex F and the medical grade hydrogel matrix in a weight ratio of 1:4;

[0041] (2) coating the mixture evenly on a non-woven fabric substrate with a thickness of 0.2-0.3 mm;

[0042] (3) Dry at 40-45°C for 4-6 hours;

[0043] (4) Cut the prepared sleeping patch into pieces and seal them.

[0044] Compared with the prior art, the present invention has the following beneficial technical effects:

[0045] 1. Multi-target synergistic effect: Linalool and α-pinene in the present invention enhance the function of GABAA receptors, while citral and geraniol regulate the 5-HT system to achieve comprehensive regulation of the sleep-wake cycle. Experimental results show that the present invention significantly improves the levels of GABA and 5-HT in the brain, shortens the sleep latency, and prolongs the sleep time; Chinese date seed oil contains a large amount of flavonoids and triterpenoids, which have sedative, sleep-inducing and anti-anxiety effects; bergamot oil is rich in limonene and linalyl acetate, which has sedative, anti-depressant and mood-improving effects. The addition of these two essential oils will produce a synergistic effect with the original ingredients, further enhancing the multi-target tranquilizing and sleep-inducing effects of the complex of the present invention, and may also bring additional anti-anxiety and anti-depressant effects, providing more comprehensive improvements for insomnia patients.

[0046] 2. Highly efficient transdermal absorption: By introducing carvacrol as a natural transdermal enhancer and using β-cyclodextrin inclusion technology, the present invention greatly improves the transdermal absorption efficiency of the active ingredients. Transdermal absorption experiments show that the 24-hour cumulative absorption of the main active ingredients is significantly higher than that of the control group.

[0047] 3. Long-term sustained release: β-cyclodextrin inclusion technology not only improves absorption efficiency, but also achieves slow release of active ingredients. The 72-hour release experiment confirms that the present invention can maintain a stable release rate, effectively solving the problem of "high at the beginning and low at the end".

[0048] 4. Excellent stability: The antioxidant effect of (R)-(-)-carvone and the introduction of soybean lecithin significantly improved the stability of the preparation. The results of the accelerated test showed that after 6 months of storage at 40℃ / 75% RH, the content retention rate of the main active ingredient was still as high as over 93%.

[0049] 5. Anti-anxiety synergistic effect: In addition to improving sleep, the present invention also exhibits significant anti-anxiety effects. The elevated plus maze test confirmed that the mice using the preparation of the present invention significantly increased their stay time in the open arm and the number of entries.

[0050] 6. Circadian rhythm regulation: The introduction of nerolidol helps regulate the expression of circadian rhythm-related genes, providing a new mechanism for improving sleep quality.

[0051] 7. Improved safety: The transdermal route of administration avoids the first-pass effect of oral administration and reduces the burden on the liver and kidneys. The sustained-release technology reduces the fluctuation of peak and valley concentrations, further reducing the risk of adverse reactions.

[0052] 8. Possibility of personalized dosing: The patch dosage form makes it possible to adjust the application area and time according to the patient's specific situation, providing a basis for personalized treatment.

[0053] In summary, the present invention has made breakthrough progress in improving sleep quality, relieving anxiety symptoms, improving bioavailability and ensuring long-term stability through the organic combination of multi-component synergy, new administration routes and advanced preparation technology. These innovations not only solve many problems in the prior art, but also provide new ideas and possibilities for the treatment of sleep disorders, and have important clinical application value and broad market prospects. DETAILED DESCRIPTION Example 1

[0054] The natural plant essential oil for calming the mind and promoting sleep in this embodiment is made of the following components by weight:

[0055] 20 parts of linalool; 15 parts of citral; 10 parts of α-pinene; 8 parts of carvacrol; 6 parts of (R)-(-)-carvone; 5 parts of nerolidol; 4 parts of geraniol; 5 parts of β-cyclodextrin; 3 parts of soybean lecithin; 1 part of vitamin E; 2 parts of jujube kernel oil; 1 part of bergamot oil; 1 part of chitosan; 10 parts of anhydrous ethanol; 5 parts of propylene glycol; 10 parts of tricaprin; 40 parts of isododecane.

[0056] The preparation method comprises the following steps:

[0057] (1) Preparation of essential oil complex:

[0058] 1.1 At 40°C, linalool, citral, α-pinene, carvacrol, (R)-(-)-carvone, nerolidol, geraniol, jujube kernel oil and bergamot oil were mixed to obtain a mixed solution A.

[0059] 1.2 Dissolve vitamin E in propylene glycol, add mixed solution A, and stir at 25°C for 2 hours to obtain solution B.

[0060] 1.3 Dissolve soybean lecithin in anhydrous ethanol, add it dropwise into solution B at a rate of 5 mL / min at 45°C, and stir for 2 hours to form microemulsion C.

[0061] 1.4 The microemulsion C was ultrasonically treated for 15 minutes (power 300 W, frequency 40 kHz) to obtain essential oil complex D.

[0062] (2) Preparation of cyclodextrin inclusion complex:

[0063] 2.1 Dissolve β-cyclodextrin in isododecane and stir at 60°C until completely dissolved.

[0064] 2.2 Add essential oil complex D into the β-cyclodextrin solution at a rate of 2 mL / min and stir for 2 hours.

[0065] 2.3 Cool the mixture to room temperature, let it stand for 12 hours, and then centrifuge (5000 rpm, 15 minutes) to collect the precipitate.

[0066] 2.4 The precipitate was vacuum dried at 40°C for 24 hours, ground and sieved (100 mesh) to obtain cyclodextrin inclusion complex E.

[0067] (3) Preparation of the final complex:

[0068] 3.1 The essential oil complex D and the cyclodextrin inclusion complex E were mixed in a weight ratio of 75:8 and stirred at 50°C for 1 hour.

[0069] 3.2 Add chitosan pre-dissolved in tricaprin and stir for 30 minutes.

[0070] 3.3 Cool the mixture to 30℃, add the remaining isododecane and stir well.

[0071] 3.4 The mixture was homogenized under vacuum at 0.05 MPa for 10 min to obtain the final essential oil complex F. Example 2

[0072] The natural plant essential oil for calming the mind and promoting sleep in this embodiment is made of the following components by weight:

[0073] 22 parts of linalool; 17 parts of citral; 12 parts of α-pinene; 10 parts of carvacrol; 8 parts of (R)-(-)-carvone; 7 parts of nerolidol; 6 parts of geraniol; 7 parts of β-cyclodextrin; 5 parts of soybean lecithin; 2 parts of vitamin E; 4 parts of jujube kernel oil; 2 parts of bergamot oil; 2 parts of chitosan; 12 parts of anhydrous ethanol; 6 parts of propylene glycol; 12 parts of tricaprin; 45 parts of isododecane.

[0074] The preparation method comprises the following steps:

[0075] (1) Preparation of essential oil complex:

[0076] 1.1 At 42°C, linalool, citral, α-pinene, carvacrol, (R)-(-)-carvone, nerolidol, geraniol, jujube kernel oil and bergamot oil were mixed to obtain a mixed solution A.

[0077] 1.2 Dissolve vitamin E in propylene glycol, add mixed solution A, and stir at 27°C for 2 hours to obtain solution B.

[0078] 1.3 Dissolve soybean lecithin in anhydrous ethanol and add it dropwise into solution B at a rate of 6 mL / min at 47°C. Stir for 2 hours to form microemulsion C.

[0079] 1.4 The microemulsion C was ultrasonically treated for 17 minutes (power 300 W, frequency 40 kHz) to obtain essential oil complex D.

[0080] (2) Preparation of cyclodextrin inclusion complex:

[0081] 2.1 Dissolve β-cyclodextrin in isododecane and stir at 62°C until completely dissolved.

[0082] 2.2 Add essential oil complex D into the β-cyclodextrin solution at a rate of 2.5 mL / min and stir for 2 hours.

[0083] 2.3 Cool the mixture to room temperature, let it stand for 12 hours, and then centrifuge (5000 rpm, 15 minutes) to collect the precipitate.

[0084] 2.4 The precipitate was vacuum dried at 42°C for 24 hours, ground and sieved (100 mesh) to obtain cyclodextrin inclusion complex E.

[0085] (3) Preparation of the final complex:

[0086] 3.1 The essential oil complex D and the cyclodextrin inclusion complex E were mixed in a weight ratio of 75:8 and stirred at 52°C for 1 hour.

[0087] 3.2 Add chitosan pre-dissolved in tricaprin and stir for 30 minutes.

[0088] 3.3 Cool the mixture to 32°C, add the remaining isododecane and stir well.

[0089] 3.4 After homogenization for 12 minutes under vacuum conditions of 0.07 MPa, the final essential oil complex F was obtained. Example 3

[0090] The natural plant essential oil for calming the mind and promoting sleep in this embodiment is made of the following components by weight:

[0091] 25 parts of linalool; 20 parts of citral; 15 parts of α-pinene; 12 parts of carvacrol; 10 parts of (R)-(-)-carvone; 9 parts of nerolidol; 8 parts of geraniol; 10 parts of β-cyclodextrin; 7 parts of soybean lecithin; 3 parts of vitamin E; 6 parts of jujube kernel oil; 4 parts of bergamot oil; 3 parts of chitosan; 15 parts of anhydrous ethanol; 8 parts of propylene glycol; 15 parts of tricaprin; 50 parts of isododecane.

[0092] The preparation method comprises the following steps:

[0093] (1) Preparation of essential oil complex:

[0094] 1.1 At 45°C, linalool, citral, α-pinene, carvacrol, (R)-(-)-carvone, nerolidol, geraniol, jujube kernel oil and bergamot oil were mixed to obtain a mixed solution A.

[0095] 1.2 Dissolve vitamin E in propylene glycol, add mixed solution A, and stir at 30°C for 2 hours to obtain solution B.

[0096] 1.3 Dissolve soybean lecithin in anhydrous ethanol and add it dropwise into solution B at a rate of 7 mL / min at 50°C. Stir for 2 hours to form microemulsion C.

[0097] 1.4 The microemulsion C was ultrasonically treated for 20 minutes (power 300 W, frequency 40 kHz) to obtain essential oil complex D.

[0098] (2) Preparation of cyclodextrin inclusion complex:

[0099] 2.1 Dissolve β-cyclodextrin in isododecane and stir at 65°C until completely dissolved.

[0100] 2.2 Add essential oil complex D into the β-cyclodextrin solution at a rate of 3 mL / min and stir for 2 hours.

[0101] 2.3 Cool the mixture to room temperature, let it stand for 12 hours, and then centrifuge (5000 rpm, 15 minutes) to collect the precipitate.

[0102] 2.4 The precipitate was vacuum dried at 45°C for 24 hours, ground and sieved (100 mesh sieve) to obtain cyclodextrin inclusion complex E.

[0103] (3) Preparation of the final complex:

[0104] 3.1 The essential oil complex D and the cyclodextrin inclusion complex E were mixed in a weight ratio of 75:8 and stirred at 55°C for 1 hour.

[0105] 3.2 Add chitosan pre-dissolved in tricaprin and stir for 30 minutes.

[0106] 3.3 Cool the mixture to 35°C, add the remaining isododecane and stir well.

[0107] 3.4 The mixture was homogenized under vacuum at 0.1 MPa for 15 minutes to obtain the final essential oil complex F. Example 4

[0108] The natural plant essential oil for calming the mind and promoting sleep in this embodiment is made of the following components by weight:

[0109] 23 parts of linalool; 18 parts of citral; 13 parts of α-pinene; 11 parts of carvacrol; 9 parts of (R)-(-)-carvone; 8 parts of nerolidol; 7 parts of geraniol; 8 parts of β-cyclodextrin; 6 parts of soybean lecithin; 2.5 parts of vitamin E; 5 parts of jujube kernel oil; 3 parts of bergamot oil; 2.5 parts of chitosan; 13 parts of anhydrous ethanol; 7 parts of propylene glycol; 13 parts of tricaprin; 47 parts of isododecane.

[0110] The preparation method comprises the following steps:

[0111] (1) Preparation of essential oil complex:

[0112] 1.1 At 43°C, linalool, citral, α-pinene, carvacrol, (R)-(-)-carvone, nerolidol, geraniol, jujube kernel oil and bergamot oil were mixed to obtain a mixed solution A.

[0113] 1.2 Dissolve vitamin E in propylene glycol, add mixed solution A, and stir at 28°C for 2 hours to obtain solution B.

[0114] 1.3 Dissolve soybean lecithin in anhydrous ethanol, add it dropwise into solution B at a rate of 6.5 mL / min at 48°C, and stir for 2 hours to form microemulsion C.

[0115] 1.4 The microemulsion C was ultrasonically treated for 18 minutes (power 300 W, frequency 40 kHz) to obtain essential oil complex D.

[0116] (2) Preparation of cyclodextrin inclusion complex:

[0117] 2.1 Dissolve β-cyclodextrin in isododecane and stir at 63°C until completely dissolved.

[0118] 2.2 Add essential oil complex D into the β-cyclodextrin solution at a rate of 2.7 mL / min and stir for 2 hours.

[0119] 2.3 Cool the mixture to room temperature, let it stand for 12 hours, and then centrifuge (5000 rpm, 15 minutes) to collect the precipitate.

[0120] 2.4 The precipitate was vacuum dried at 43°C for 24 hours, ground and sieved (100 mesh) to obtain cyclodextrin inclusion complex E.

[0121] (3) Preparation of the final complex:

[0122] 3.1 The essential oil complex D and the cyclodextrin inclusion complex E were mixed in a weight ratio of 75:8 and stirred at 53°C for 1 hour.

[0123] 3.2 Add chitosan pre-dissolved in tricaprin and stir for 30 minutes.

[0124] 3.3 Cool the mixture to 33°C, add the remaining isododecane and stir well.

[0125] 3.4 After homogenization for 13 minutes under vacuum conditions of 0.08 MPa, the final essential oil complex F was obtained.

[0126] Comparative Example 1, a composite lacking α-pinene (corresponding to Example 1)

[0127] The composition of the composite of this comparative example is the same as that of Example 1, but does not contain α-pinene. The weight parts of other components remain unchanged.

[0128] The preparation method is the same as that of Example 1, but α-pinene is not added in step 1.1.

[0129] Comparative Example 2, a complex in which (R)-(-)-carvone is replaced by menthone (corresponding to Example 2)

[0130] The composition of the composite of this comparative example is the same as that of Example 2, except that menthone (8 parts) is used to replace (R)-(-)-carvone. The weight parts of other components remain unchanged.

[0131] The preparation method is the same as in Example 2, except that menthone is used instead of (R)-(-)-carvone in step 1.1.

[0132] Comparative Example 3, a composite with too high linalool content (corresponding to Example 3)

[0133] The composition of the composite of this comparative example is the same as that of Example 3, but the content of linalool is increased to 30 parts. The weight parts of other components remain unchanged.

[0134] The preparation method is the same as that of Example 3, but in step 1.1, the amount of linalool added is 30 parts.

[0135] Comparative Example 4: Complex lacking β-cyclodextrin inclusion (corresponding to Example 4)

[0136] The composition of the composite of this comparative example is the same as that of Example 4, but does not contain β-cyclodextrin, and the cyclodextrin inclusion step is not performed.

[0137] The preparation method comprises the following steps:

[0138] (1) Preparation of essential oil complex:

[0139] Steps 1.1 to 1.4 are the same as in Example 4.

[0140] (2) Preparation of the final complex:

[0141] 2.1 Essential oil complex D was mixed with soybean lecithin (pre-dissolved in tricaprin, weight ratio 1:5) and stirred at 53 °C for 1 hour.

[0142] 2.2 Add chitosan pre-dissolved in tricaprin and stir for 30 minutes.

[0143] 2.3 Cool the mixture to 33°C, add isododecane and stir evenly.

[0144] 2.4 After homogenization for 13 minutes under vacuum conditions of 0.08 MPa, the final essential oil complex F was obtained.

[0145] Comparative Example 5: Composite with too low citral content (corresponding to Example 2)

[0146] The composition of the composite of this comparative example is the same as that of Example 2, but the content of citral is reduced to 10 parts. The weight parts of other components remain unchanged.

[0147] The preparation method is the same as that of Example 2, but the amount of citral added in step 1.1 is 10 parts.

[0148] Comparative Example 6: Complex lacking soybean lecithin (corresponding to Example 3)

[0149] The composition of the composite of this comparative example is the same as that of Example 3, but does not contain soybean lecithin. The weight portions of other components remain unchanged.

[0150] The preparation method is the same as that of Example 3, but soybean lecithin is not added in step 1.3, and step 1.4 is directly performed.

[0151] In order to study the mechanism of action of the present invention, the applicant has designed a number of evaluation models and provided experimental conditions, method steps and detailed test results.

[0152] The evaluation model is designed as follows:

[0153] 1. Transdermal absorption model: evaluate the transdermal permeability of the complex;

[0154] 2. Sleep quality assessment model: evaluate the effect of the compound on sleep latency and sleep time;

[0155] 3. Anxiolytic model: evaluate the anxiolytic effect of the compound;

[0156] 4. Determination of neurotransmitter levels: evaluate the effect of the complex on GABA and melatonin levels;

[0157] 5. Sustained release performance evaluation: evaluate the sustained release time of the complex;

[0158] 6. Stability test: Evaluate the stability of the compound under different temperature and humidity conditions.

[0159] Experimental methods and steps:

[0160] 1. Transdermal absorption model (Franz diffusion cell method): Using ex vivo porcine skin as a skin model, 0.5 g of sample was added to the supply chamber of the Franz diffusion cell and PBS buffer (pH 7.4) was filled in the receiving chamber. A 24-hour permeation experiment was performed at 37°C and samples were taken every 4 hours. The cumulative permeation amount of the main active ingredient was determined by HPLC.

[0161] 2. Sleep quality assessment model (sodium pentobarbital-induced sleep experiment in mice): 6-8 week old male ICR mice were selected, 10 in each group; mice were given a transdermal patch (2 cm × 2 cm) for 7 days; on the evening of the 7th day, mice were given 45 mg / kg sodium pentobarbital (intraperitoneal injection); the time from injection to loss of righting reflex (sleep latency) and the time to restore righting reflex (sleep time) were recorded.

[0162] 3. Anxiety-relief model (mouse elevated plus maze test): 6-8 week old male ICR mice were selected, 10 per group; mice were given a transdermal patch (2 cm × 2 cm) for 7 days; on the 7th day, the mice were placed in an elevated plus maze for 5 minutes; the time the mice stayed in the open arm and the number of entries were recorded.

[0163] 4. Determination of neurotransmitter levels: After the above-mentioned mouse experiment was completed, the mice were immediately killed and the brains were removed; the hippocampus and cortex regions were separated and tissue homogenates were prepared; and the GABA and 5-HT levels were determined using the ELISA method.

[0164] 5. Evaluation of sustained-release performance: Use the receiving solution of the transdermal absorption model; take samples every 4 hours for 72 hours; use HPLC to determine the release curves of linalool, citral, α-pinene, carvacrol and (R)-(-)-carvone.

[0165] 6. Stability test: The samples were stored at 25°C / 60% RH and 40°C / 75% RH for 6 months. Samples were taken every month to test the content of the main active ingredients and changes in appearance.

[0166] 7. Evaluation of circadian gene expression: 6-8 week old male ICR mice were selected, 10 mice in each group; mice were given a transdermal patch (2 cm × 2 cm) for 7 days; samples were taken at different time points on the 7th day (such as ZT0, ZT6, ZT12, and ZT18); total RNA from hippocampal and hypothalamic tissues was extracted; and the expression levels of circadian rhythm-related genes such as Clock, Bmal1, Per1, and Per2 were determined using real-time quantitative PCR.

[0167] 8. Evaluation of anti-inflammatory effect (new): 6-8 week old male ICR mice were selected, 10 mice in each group; mice were given a transdermal patch (2 cm × 2 cm) for 7 days; on the 7th day, lipopolysaccharide (LPS, 1 mg / kg) was intraperitoneally injected to induce an inflammatory response, and blood samples were collected 6 hours later. The levels of TNF-α, IL-6, and IL-1β in the serum were determined by ELISA.

[0168] The experimental results are shown in Table 1-8:

[0169] Table 1. Cumulative transdermal absorption of main active ingredients in 24 hours (μg / cm²)

[0170]

[0171] Table 2. Sleep quality assessment results

[0172]

[0173] Table 3. Results of the anxiolytic model

[0174]

[0175] Table 4. Results of neurotransmitter level determination (pg / mg protein)

[0176]

[0177] Table 5. Sustained release performance evaluation results (72-hour cumulative release percentage of the main active ingredient)

[0178]

[0179] Table 6. Stability test results (main active ingredient content retention after 6 months)

[0180]

[0181] Table 7. Relative expression levels of major circadian rhythm genes (GAPDH was used as the internal reference, and the expression level of the blank control group at ZT0 was set to 1)

[0182]

[0183] Table 8. Serum inflammatory factor levels (pg / mL)

[0184]

[0185] Results and discussion:

[0186] 1. Transdermal absorption: Examples 1-4 all showed excellent transdermal absorption performance, among which Example 3 had the best effect. The transdermal absorption effects of Comparative Example 4 (lack of β-cyclodextrin inclusion) and Comparative Example 5 (too low content of menthone) were significantly reduced, indicating that β-cyclodextrin inclusion and an appropriate amount of menthone are crucial to improving transdermal absorption.

[0187] 2. Sleep quality: Examples 1-4 all significantly shortened the sleep latency and prolonged the sleep time, with Example 3 having the best effect. The effect of Comparative Example 1 (lacking α-pinene) was significantly lower than that of the other samples, confirming the importance of α-pinene as a new sleep regulator.

[0188] 3. Anti-anxiety effect: Examples 1-4 showed significant anti-anxiety effects in the elevated plus maze test, increasing the mice's stay time and number of entries in the open arms. The effect of Comparative Example 2 (replacing (R)-(-)-carvone with menthone) was slightly worse, indicating that (R)-(-)-carvone has a unique effect in anti-anxiety.

[0189] 4. Neurotransmitter levels: Examples 1-4 significantly increased the levels of GABA and melatonin in the mouse brain, with Example 3 having the best effect. Comparative Example 1 (lacking α-pinene) and Comparative Example 4 (lacking β-cyclodextrin inclusion) had poor neurotransmitter level enhancement effects, further confirming the importance of α-pinene and β-cyclodextrin inclusion.

[0190] 5. Sustained release performance: Examples 1-4 showed excellent sustained release performance, with the cumulative release rate of each main active ingredient being above 85% within 72 hours. The sustained release effect of Comparative Example 4 (lacking β-cyclodextrin inclusion) was significantly reduced, with a cumulative release rate of only 70-80%, which fully demonstrated the key role of β-cyclodextrin inclusion in achieving the sustained release effect.

[0191] 6. Stability: Examples 1-4 all showed good stability under different temperature and humidity conditions. After 6 months, the content retention rate of the main active ingredients was higher than 97% under normal temperature conditions and remained above 93% under accelerated test conditions. The stability of Comparative Example 6 (lacking soybean lecithin) was slightly worse, indicating that soybean lecithin played an important role in improving the stability of the composite.

[0192] 7. Circadian rhythm gene expression: Examples 1-4 all significantly increased the expression levels of major circadian rhythm genes (Clock, Bmal1, Per1, Per2), with Example 3 having the most significant effect. This indicates that the complex of the present invention can effectively regulate the expression of biological clock genes and help to reestablish normal circadian rhythms.

[0193] The gene expression levels of Comparative Example 1 (lacking α-pinene) and Comparative Example 4 (lacking β-cyclodextrin inclusion) were significantly lower than those of the embodiment, indicating that α-pinene and β-cyclodextrin inclusion are essential for maintaining the biological activity of the complex. This may be due to the synergistic effect of α-pinene and the improved bioavailability of the active ingredient by β-cyclodextrin inclusion.

[0194] Comparative Example 5 (too low citral content) also showed a weaker gene regulation effect, further confirming the important role of citral in regulating circadian rhythms. This is consistent with previous studies reporting the effects of citral on the biological clock.

[0195] 8. Anti-inflammatory effect: Examples 1-4 all significantly reduced the levels of LPS-induced inflammatory factors (TNF-α, IL-6, IL-1β), among which Example 3 had the most significant anti-inflammatory effect. This indicates that the complex of the present invention has a strong anti-inflammatory effect and may help alleviate the inflammatory response associated with sleep disorders.

[0196] Comparative Example 2 (replacing (R)-(-)-carvone with menthone) showed the weakest anti-inflammatory effect, confirming the unique anti-inflammatory effect of (R)-(-)-carvone in this complex. This is consistent with the anti-inflammatory properties of (R)-(-)-carvone reported in previous studies.

[0197] The anti-inflammatory effect of Comparative Example 6 (lacking soybean lecithin) is slightly lower than that of the example, indicating that soybean lecithin not only improves the stability of the complex, but also indirectly enhances the anti-inflammatory effect by enhancing the bioavailability of the active ingredient.

[0198] The present invention has the following unexpected technical effects:

[0199] 1. Synergistic effect: The present invention achieves multi-target synergistic effect through a carefully designed multi-component formula. The experimental results show that there is a significant synergistic effect between the components, making the overall effect far superior to the effect of a single component or simple superposition. For example, the combination of α-pinene and linalool not only increases the level of GABA, but also significantly increases the secretion of melatonin. This dual effect is relatively rare in the prior art.

[0200] 2. Balance between transdermal absorption and sustained release: The present invention achieves long-term sustained release while improving transdermal absorption through β-cyclodextrin inclusion technology and the addition of an appropriate amount of menthone. This balance solves the technical contradiction of traditional essential oil preparations, which are either insufficient absorption or too fast release.

[0201] 3. Synergy of multiple drug effects: The present invention not only improves sleep quality, but also has a significant anti-anxiety effect. This synergistic effect of multiple drugs provides a more comprehensive treatment plan for patients with sleep disorders, especially for insomnia patients with anxiety symptoms.

[0202] 4. Breakthrough in stability: By introducing soybean lecithin and β-cyclodextrin inclusion technology, the present invention not only ensures the stability of the active ingredients, but also maintains good transdermal absorption performance. This breakthrough solves the problem of the difficulty in balancing the stability and bioavailability of traditional essential oil preparations.

[0203] 5. Possibility of individualized administration: Since the present invention adopts the form of a transdermal patch, the patch size or application time can be adjusted according to the specific conditions of the patient, providing the possibility for individualized treatment. This flexibility is difficult to achieve in oral preparations.

[0204] 6. Improved safety: Through sustained-release technology and transdermal administration, the present invention reduces the peak concentration of active ingredients and reduces the risk of potential side effects. At the same time, it avoids the first-pass effect of oral administration and improves the bioavailability of the drug.

[0205] 7. Multiple mechanisms of action: The complex of the present invention exhibits multiple mechanisms of action by regulating circadian gene expression and inhibiting inflammatory responses. This multi-target synergistic effect may be the key to its superiority over a single component or a simple mixture. The excellent performance of Example 3 shows that precise regulation of the proportions of each component is crucial to achieving the best effect. The synergistic effect of key ingredients such as α-pinene, (R)-(-)-carvone, and citral not only enhances the sleep improvement effect, but also significantly improves the anti-inflammatory ability.

[0206] 8. Circadian rhythm regulation: The addition of nerolidol may be a key factor in improving circadian rhythm gene expression. This discovery provides a new mechanism for improving sleep quality and is expected to solve the problem of insufficient regulation of the biological clock in existing sleep disorder treatments.

[0207] 9. New anti-inflammatory pathway: The potent anti-inflammatory effect of (R)-(-)-carvone provides a new approach to the treatment of sleep disorders. By reducing the inflammatory response, it may help break the vicious cycle between sleep disorders and chronic inflammation.

[0208] 10. Importance of formulation technology: The application of β-cyclodextrin inclusion and soybean lecithin not only improves the stability and bioavailability of the complex, but also indirectly enhances its biological effect. This emphasizes the important role of advanced formulation technology in realizing the potential of natural products.

[0209] 11. Potential for individualized treatment: The differences in effects between different embodiments suggest that the needs of different patients can be met by adjusting the proportion of components, providing the possibility for individualized treatment of sleep disorders.

[0210] 12. Safety advantage: Compared with traditional sedative hypnotic drugs, the multi-target mild regulatory effect of the complex of the present invention may bring better safety. The addition of anti-inflammatory effect may help reduce potential side effects caused by long-term use.

[0211] 13. Broad-spectrum therapeutic prospects: By simultaneously regulating circadian rhythm and inhibiting inflammation, the complex of the present invention may not only be suitable for primary insomnia, but may also have potential therapeutic effects on secondary sleep disorders associated with circadian rhythm disorders (such as shift work) and chronic inflammation.

[0212] The innovative features of the present invention are summarized as follows:

[0213] 1. Multi-component synergy: The carefully designed compound ratio achieves synergistic enhancement of circadian rhythm regulation and anti-inflammatory effects.

[0214] 2. New target of circadian rhythm: Nerolidol was used for the first time to treat sleep disorders, opening up a new way to regulate the biological clock.

[0215] 3. New anti-inflammatory strategy: The introduction of (R)-(-)-carvone enhances the anti-inflammatory effect and provides a new mechanism of action for the treatment of sleep disorders.

[0216] 4. Advanced formulation technology: The innovative application of β-cyclodextrin inclusion and liposome technology significantly improves the biological effect of the complex.

[0217] 5. Basis for individualized treatment: Implementation examples with different ratios lay the foundation for the future development of individualized sleep disorder treatment programs.

[0218] These results not only confirm the multiple mechanisms of action of the complex of the present invention, but also reveal its unique advantages in improving sleep quality. By regulating circadian rhythm gene expression and inhibiting inflammatory response, the present invention provides a new, multi-target strategy for the treatment of sleep disorders, which is expected to overcome the limitations of existing treatment methods and provide patients with safer and more effective treatment options.

[0219] According to the comprehensive evaluation results, Example 3 performed the best and can be considered as the best embodiment of the present invention. It showed the best or near-best effects in terms of transdermal absorption, sleep quality improvement, antianxiety effect, neurotransmitter regulation, sustained release performance and stability.

[0220] The present invention has made breakthrough progress in improving sleep quality, relieving anxiety symptoms, improving bioavailability and ensuring long-term stability through the combination of multi-component synergy, new drug administration routes and advanced preparation technology. These unexpected technical effects not only solve many problems in the prior art, but also provide new ideas and possibilities for the treatment of sleep disorders. The innovation and practical value of the present invention have been fully verified in experimental data, and it is expected to bring good news to the majority of patients with sleep disorders.

[0221] Example 5

[0222] Preparation and testing of sleep patches

[0223] 1. Preparation of sleeping patch:

[0224] 1.1 Essential oil complex F was prepared according to the method of Example 3.

[0225] 1.2 Medical grade hydrogel matrix (sodium polyacrylate) and purified water were mixed in a weight ratio of 1:4 and stirred at 60°C for 2 hours to prepare a hydrogel matrix.

[0226] 1.3 The essential oil complex F and the hydrogel matrix were mixed in a weight ratio of 1:4 and stirred at 40°C for 1 hour to obtain a uniform mixture.

[0227] 1.4 Spread the mixture evenly on a non-woven fabric substrate (polyester / viscose blend, 100g / m²) with a thickness of 0.25mm.

[0228] 1.5 Dry at 42℃ for 5 hours to obtain the semi-finished sleeping patch;

[0229] 1.6 Cut the semi-finished sleeping patch into 5cm×5cm squares.

[0230] 1.7 Cover the back of the sleeping patch with a layer of silicone-coated polyethylene film as a protective layer.

[0231] 1.8 The prepared sleeping patch is sealed and packaged with an aluminum foil composite film.

[0232] 2. Testing experiment of sleeping patch:

[0233] 1. In vitro transdermal release test:

[0234] Methods: The modified Franz diffusion cell method was used. The prepared sleeping patch (5 cm × 5 cm) was fixed on the supply chamber of the diffusion cell, and the receiving chamber was filled with 40 mL of pH 7.4 PBS buffer, and the temperature was maintained at 32 ± 0.5 °C. Samples were taken at 0, 2, 4, 8, 12, 24, 48 and 72 hours, and the cumulative release of the main active ingredient was determined by HPLC.

[0235] Table 9. Cumulative release rate of the main active ingredients (%)

[0236]

[0237] 2. Skin irritation test:

[0238] Methods: Six New Zealand white rabbits were selected, and the hair on the back of each rabbit was cut to expose six 2.5 cm × 2.5 cm skin areas. Sleep patches (2.5 cm × 2.5 cm) were applied to three of the areas, and the other three areas were used as controls. Skin reactions were observed and recorded 24, 48, and 72 hours after application, and irritation was evaluated according to the Draize scoring standard.

[0239] Table 10. Skin irritation rating

[0240]

[0241] 3. Human trials:

[0242] Methods: 30 patients with mild to moderate insomnia (Pittsburgh Sleep Quality Index PSQI>5) were selected and randomly divided into an experimental group and a placebo group, with 15 people in each group. The experimental group used the sleep patch of the present invention, and the placebo group used a matrix patch without active ingredients. The subjects applied the patch to the back of the neck or the sole of the foot 30 minutes before going to bed every night for 2 consecutive weeks. The serum melatonin level and serum inflammatory factors (TNF-α, IL-6) were measured before and after 2 weeks of use.

[0243] Table 11. Human test results (pg / mL)

[0244]

[0245] *Indicates p<0.05 compared with before use, which is statistically significant.

[0246] The results are analyzed as follows:

[0247] 1. Please refer to Table 9. The in vitro transdermal release test shows that the sleeping patch can achieve sustained and slow release of the main active ingredients. The cumulative release rate of each ingredient within 72 hours is more than 90%, which meets the requirements of long-acting preparations.

[0248] 2. Please refer to Table 10. The skin irritation test results show that this sleeping patch is almost non-irritating to the skin. Only slight irritation occurs after 48 hours, and completely disappears after 72 hours, indicating that the product has good skin compatibility.

[0249] 3. Please refer to Table 11, the results of human trials show:

[0250] a) After using the sleep patch for 2 weeks, sleep quality has improved significantly.

[0251] b) The serum melatonin level in the experimental group increased significantly, confirming the promoting effect of the sleep patch on the secretion of endogenous melatonin.

[0252] c) The levels of serum inflammatory factors (TNF-α, IL-6) in the experimental group were significantly reduced, verifying the anti-inflammatory effect of the sleep patch.

[0253] d) No significant changes were observed in any of the indicators in the placebo group.

[0254] These results comprehensively prove that the sleep patch of the present invention not only has good sustained-release performance and skin compatibility, but also can effectively improve sleep quality, increase endogenous melatonin levels, and has significant anti-inflammatory effects. This multiple mechanism of action provides a safe and effective new option for insomnia patients, especially for patients who need long-term use and circadian rhythm regulation.

[0255] The above description is only an embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent substitution, improvement, etc. made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.

Claims

1. A natural plant essential oil for calming the mind and promoting sleep, characterized in that: Made of the following components by weight: 20-25 parts of linalool; 15-20 parts of citral; 10-15 parts of α-pinene; 8-12 parts of carvacrol; 6-10 parts of (R)-(-)-carvone; 5-9 parts of nerolidol; 4-8 parts of geraniol; 5-10 parts of β-cyclodextrin; 3-7 parts of soybean lecithin; 1-3 parts of vitamin E; 2-6 parts of jujube kernel oil and 1-4 parts of bergamot oil.

2. The natural plant essential oil for calming the mind and promoting sleep according to claim 1, characterized in that: It also contains the following auxiliary ingredients: 1-3 parts of chitosan; 10-15 parts of anhydrous ethanol; 5-8 parts of propylene glycol; 10-15 parts of tricaprin; 40-50 parts of isododecane.

3. The method for preparing the natural plant essential oil for calming the mind and promoting sleep according to claim 1 or 2, characterized in that: The following steps are involved: (1) preparing an essential oil complex; (2) preparing cyclodextrin inclusion complex; (3) Preparing the final complex based on the products of steps (1) and (2).

4. The method according to claim 3, characterized in that The step (1) comprises: (1.1) Mixing linalool, citral, α-pinene, carvacrol, (R)-(-)-carvone, nerolidol, geraniol, jujube kernel oil and bergamot oil at 40-45° C. to obtain a mixed solution A; (1.2) Dissolve vitamin E in propylene glycol, add mixed solution A, and stir at 25-30°C for 2 hours to obtain solution B; (1.3) Dissolve soybean lecithin in anhydrous ethanol, add it dropwise into solution B at 45-50°C, and stir for 2 hours to form microemulsion C; (1.4) The microemulsion C is ultrasonically treated for 15-20 minutes to obtain the essential oil complex D.

5. The method according to claim 3, characterized in that: The step (2) comprises: (2.1) Dissolve β-cyclodextrin in isododecane and stir at 60-65°C until completely dissolved; (2.2) Add essential oil complex D to the β-cyclodextrin solution at a rate of 2-3 mL / min and stir for 2 hours; (2.3) The mixture was cooled to room temperature, allowed to stand for 12 hours, and then centrifuged to collect the precipitate; (2.4) The precipitate was vacuum dried at 40-45°C for 24 hours, ground and sieved to obtain cyclodextrin inclusion complex E.

6. The method according to claim 3, characterized in that The step (3) comprises: (3.1) Mix the essential oil complex D and the cyclodextrin inclusion complex E and stir at 50-55°C for 1 hour; (3.2) Add chitosan pre-dissolved in tricaprin and stir for 30 minutes; (3.3) Cool the mixture to 30-35°C, add the remaining isododecane, and stir evenly; (3.4) The mixture was homogenized under a vacuum condition of 0.05-0.1 MPa for 10-15 minutes to obtain the final essential oil complex F.

7. The method according to claim 6, characterized in that The weight ratio of the essential oil complex D to the cyclodextrin inclusion complex E is 75:

8.

8. The method according to claim 6, characterized in that The weight ratio of chitosan to tricaprin is 1:

3.

9. A sleeping patch, characterized in that: The invention comprises the natural plant essential oil for calming the mind and promoting sleep as described in any one of claims 1 to 2.

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