Catnip extract and use thereof for enhancing serotonin and daytime vitality

Abstract

A catnip extract comprises one or a combination of compounds of formula I to II. The catnip extract is used to prepare compositions that shorten sleep onset time and / or improve sleep quality, enhance daytime activity, have antidepressant effects, or protect the brain. The catnip extract is obtained by extracting catnip flower spikes with water at 85±5°C.

Description

Catnip extract and its uses in improving sleep and combating depression This invention relates to a catnip extract, and more particularly to a catnip extract and its use in improving sleep and combating depression. Sleep plays a vital role in human health and beauty, allowing the body and skin to fully rest and recover. Insomnia has many causes, potentially related to stress, emotions, lifestyle habits, illness, and medications. Insomnia not only harms physical and mental health but also increases the risk of various diseases. Since the rise of organic and / or natural dietary concepts, biotechnology companies and food manufacturers have actively invested in the research and development of products related to natural plants. To ensure that the health benefits of plant-based products are scientifically validated, the analysis of active plant components and the evaluation of their efficacy have become key aspects of product development. catnip Nepeta cataria L. is a plant of the Lamiaceae family with a distinctive odor. Biotechnology companies and food manufacturers are actively researching the active ingredients and efficacy of catnip to develop related products. In view of the above, in some embodiments, a catnip extract is used to prepare compositions that shorten sleep onset time and / or improve sleep quality. The catnip extract is obtained by extracting catnip flower spikes with water at 85±5°C. In some embodiments, catnip extract has the effect of increasing melatonin levels. In some embodiments, catnip extract has the effect of reducing the proportion of light sleep and / or increasing the proportion of deep sleep. In some embodiments, a catnip extract is used to prepare a composition for enhancing daytime activity. The catnip extract is obtained by extracting catnip flower spikes with water at 85±5°C. In some embodiments, a catnip extract is used to prepare an antidepressant composition. The catnip extract is obtained by extracting catnip flower spikes with water at 85±5°C. In some embodiments, catnip extract has the effect of increasing serotonin levels. In some embodiments, catnip extract has the effect of reducing nerve overactivation. In some embodiments, a catnip extract is used to prepare a brain-protective composition. The catnip extract is obtained by extracting catnip flower spikes with water at 85±5°C. In some embodiments, catnip extract has the effect of enhancing the antioxidant capacity of nerve cells. In some embodiments, a catnip extract comprises one or a combination of compounds of formula I to formula II. In summary, the use of catnip extract in improving sleep and antidepressant effects in any embodiment relates to the use of catnip extract in preparing a composition for improving sleep or antidepressant effects, thereby providing a composition that, when applied to an individual, produces an effect of improving sleep or antidepressant effects in that individual. In other words, the aforementioned composition has the function of improving sleep or antidepressant effects. In some embodiments, the composition obtained from catnip extract also has one or more of the following functions: shortening sleep onset time and / or improving sleep quality, increasing daytime activity energy, antidepressant effects, and brain protection. In some embodiments, catnip extract is made from catnip ( The catnip was obtained by extraction using Nepeta cataria L. as a raw material. The extraction process mainly involves extracting catnip with a solvent to dissolve the active ingredients in the catnip into the solvent. In some embodiments, the catnip being extracted is a catnip flower spike. In some embodiments, the catnip flower spike being extracted includes a corolla, calyx, receptacle, and stamens. In some embodiments, the catnip being extracted may be a whole, untreated flower spike (i.e., the corolla, calyx, receptacle, and stamens are not separated and their size has not been physically pretreated), or it may be in a form that has been physically pretreated and broken down into small pieces, granules, or powder. The physical pretreatment may include at least one of the following: coarse crushing, chopping, shearing, mashing, and grinding. In some embodiments, the catnip being extracted is a whole, untreated flower spike. In some embodiments, the flower spikes to be extracted may be freshly collected flower spikes, dried flower spikes, and / or frozen flower spikes. For example, in the extraction process, dried flower spikes are extracted using a solvent. In some embodiments, the extraction process includes extracting catnip with water at 85±5°C for 60 to 90 minutes to obtain a preliminary extract. For example, catnip can be soaked in water at 85±5°C for 60 minutes to dissolve the active ingredients in the catnip into the water to obtain a preliminary extract. In some embodiments, during the extraction process, the solvent is water, the raw material is catnip, and the initial weight ratio of the solvent to the raw material is 10-20:1. For example, the weight ratio of water to catnip is 16:1. In some embodiments, during the extraction process, the initial extract may be further filtered to remove solids such as catnip from the water extraction, resulting in a filtrate. For example, the initial extract may be filtered through a 400-mesh filter to remove solids, and the filtrate may be collected. In some embodiments, the filtrate may be further concentrated during the extraction process to obtain a concentrate. In some embodiments, the filtrate may be concentrated under reduced pressure at 55°C-65°C to obtain a concentrate. For example, the filtrate may be concentrated under reduced pressure at 60±5°C. In some embodiments, the concentration time may be determined by the Brix value of the concentrate, but is not limited thereto. Continuing with the previous example, the Brix value of the obtained concentrate is 7.5±0.5°Bx. In other words, the filtrate may be concentrated under reduced pressure at 60±5°C until the concentrated filtrate has a Brix value of 7.5±0.5°Bx, and the concentrated filtrate at this point is the concentrate. In some embodiments, during the extraction process, the initial extract may be concentrated to form a concentrated solution with a reduced liquid volume. The concentrated solution is then filtered to remove solids, and the filtrate is collected. In some embodiments, the initial extract is either concentrated or filtered during the extraction process. It should be understood that the initial extract, filtrate, or concentrate obtained from the extraction process can be used as catnip extract, depending on actual needs. In some embodiments, the aforementioned catnip extract has the ability to shorten sleep onset time and / or improve sleep quality. Therefore, catnip extract is suitable for preparing compositions that shorten sleep onset time and / or improve sleep quality. In some embodiments, catnip extract has the ability to increase melatonin levels. In other words, when catnip extract is administered to an individual, it can increase that individual's melatonin levels. Therefore, catnip extract is suitable for preparing compositions that increase melatonin levels. In some embodiments, catnip extract has the ability to reduce the proportion of light sleep and / or increase the proportion of deep sleep. In other words, when catnip extract is administered to an individual, it can reduce the proportion of light sleep and / or increase the proportion of deep sleep in that individual. Therefore, catnip extract is suitable for preparing compositions that reduce the proportion of light sleep and / or increase the proportion of deep sleep. In some embodiments, the aforementioned catnip extract has the ability to enhance daytime activity levels. Therefore, catnip extract is suitable for preparing compositions that enhance daytime activity levels. In some embodiments, the aforementioned catnip extract has antidepressant properties. Therefore, catnip extract is suitable for preparing antidepressant compositions. In some embodiments, catnip extract has the ability to increase serotonin levels. In other words, when catnip extract is administered to an individual, it can increase that individual's serotonin levels. Therefore, catnip extract is suitable for preparing compositions that increase serotonin levels. In some embodiments, catnip extract has the ability to reduce neural hyperactivation. In other words, when catnip extract is administered to an individual, it reduces neural hyperactivation in that individual. Therefore, catnip extract is suitable for preparing compositions that reduce neural hyperactivation. In some embodiments, the aforementioned catnip extract has brain-protective properties. Therefore, catnip extract is suitable for preparing brain-protective compositions. In some embodiments, catnip extract has the ability to enhance the antioxidant capacity of nerve cells. In other words, when catnip extract is administered to an individual, it can enhance the antioxidant capacity of that individual's nerve cells. Therefore, catnip extract is suitable for preparing compositions that enhance the antioxidant capacity of nerve cells. In some embodiments, the catnip extract comprises one or a combination of compounds of formula I to II. In some embodiments, the compound of Formula I is (2S,3S,4S,5R,6S)-6-[5-(5,7-dihydroxy-4-oxo-4H-benzopyran-2-yl)-2-hydroxyphenoxy]-3,4,5-trihydroxyoxane-2-carboxylic acid. In some embodiments, the compound of Formula I is luteolin-3'-glucoronide. Oleacetin-3'-gluconic acid is the common name for (2S,3S,4S,5R,6S)-6-[5-(5,7-dihydroxy-4-oxo-4H-benzopyran-2-yl)-2-hydroxyphenoxy]-3,4,5-trihydroxyoxane-2-carboxylic acid. In some embodiments, the compound of formula II is 7a-hydroxy-3,6-dimethyl-2,4,5,6,7,7a-hexahydrocyclohexa[1,2-b]furan-2-one. In some embodiments, the aforementioned individual may be a person. In some embodiments, the daily intake of catnip extract in the aforementioned composition is 3 grams. In some embodiments, the prepared composition may be a pharmaceutical composition or an edible composition for non-medical purposes. In some embodiments, when the aforementioned composition is a pharmaceutical composition, the pharmaceutical composition comprises an effective amount of catnip extract. This pharmaceutical composition can be manufactured using techniques known to those skilled in the art into dosage forms suitable for enteral, parenterally, orally, or topically administration. In some embodiments, the dosage form for oral or enteral administration may be, but is not limited to, tablets, troche, lozenges, pills, capsules, dispersible powders, granules, solutions, suspensions, emulsions, syrups, elixirs, slurries, or similar substances. In some embodiments, the dosage form for non-enteral or local administration may be, but is not limited to, an injection [e.g., a sterile aqueous solution or dispersion], a sterile powder, an external preparation, or the like. In some embodiments, the administration method of the injectable may be, but is not limited to, intraperitoneal injection, subcutaneous injection, intraepidermal injection, intradermal injection, intramuscular injection, intravenous injection, or intralesional injection. In some embodiments, a pharmaceutical composition containing an effective amount of catnip extract may further comprise a pharmaceutically acceptable carrier widely used in pharmaceutical manufacturing techniques. In some embodiments, a pharmaceutically acceptable carrier may be one or more of the following: solvent, buffer, emulsifier, suspending agent, decomposer, disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, wetting agent, lubricant, absorption delaying agent, liposome, and the like. The type and quantity of carrier selected fall within the scope of professional competence and routine technique of a person skilled in this art. Among them, the solvents that can be used as pharmaceutically acceptable carriers are water, normal saline, phosphate buffered saline (PBS), or an aqueous solution containing alcohol. In some embodiments, a pharmaceutical composition containing an effective amount of catnip extract can be manufactured into a topical preparation suitable for application to the skin using techniques known to those skilled in the art. In some embodiments, the topical preparation includes, but is not limited to: emulsion, gel, ointment, cream, patch, liniment, powder, aerosol, spray, lotion, serum, paste, foam, drop, suspension, salve, and bandage. In some embodiments, when the aforementioned pharmaceutical composition is an external formulation, the pharmaceutical composition may be prepared by mixing an effective amount of catnip extract with a base known to those skilled in the art. In some embodiments, the substrate may contain one or more additives selected from the following: water, alcohols, glycols, hydrocarbons [such as petroleum jelly and white petrolatum], waxes [such as paraffin and yellow wax], preserving agents, antioxidants, surfactants, absorption enhancers, stabilizing agents, gelling agents [such as Carbopol® 974P, microcrystalline cellulose, and carboxymethylcellulose], active agents, humectants, odor absorbers, fragrances, pH adjusting agents, and chelating agents. The additives include agents, emulsifiers, occlusive agents, softeners, thickeners, solubilizing agents, penetration enhancers, anti-irritants, colorants, and propellants. The selection and quantity of these additives fall within the scope of professional expertise and routine practice of those familiar with this technology. In some embodiments, when the aforementioned composition is an edible composition for non-medical purposes, the edible composition contains a specific amount of catnip extract. The edible composition may be in the form of powder, granules, solution, colloid, or paste. In some embodiments, edible compositions containing catnip extract for non-medical purposes may be food products or food additives. In some embodiments, the edible composition containing catnip extract may be a beverage, fermented food, bakery product, health food, or dietary supplement. In some embodiments, the edible composition containing catnip extract may further include an adjuvant. For example, the adjuvant may be maltodextrin, malic acid, sucralose, citric acid, fruit flavoring, honey flavoring, steviol glycosides, or combinations thereof. The type and quantity of the adjuvant selected fall within the scope of the expertise and routine practice of those skilled in the art. In some embodiments, food additives may be seasonings, sweeteners, flavorings, pH adjusters, emulsifiers, colorants, or stabilizers, etc. Unless otherwise specified, the experimental procedures in the following examples are performed at room temperature (25℃-30℃) and normal pressure (1 atm). Example 1: Preparation of catnip extract A. Materials: 1. Catnip (Nepeta cataria L.) flower spikes (origin: China). 2. Secondary water, also known as RO water (Reverse Osmosis) or double-distilled water, hereinafter referred to as "water". B. Preparation process: 1. Heat water to 85±5℃, add catnip flower spikes, and soak the catnip flower spikes in the 85±5℃ water for 60 minutes to form a primary extract containing solids. The weight ratio of catnip powder to water is 1:16. 2. Filter the cooled initial extract through a 400-mesh filter to remove solids (i.e., the extracted catnip flower spikes) to obtain the filtrate. 3. Set the temperature of the concentrator (model: Rotavapor R-100; brand: BUCHI) to 60±5℃, and then use the concentrator to concentrate the filtrate under reduced pressure. Stop the concentration when the Brix value of the filtrate reaches 7.5±0.5°Bx, and the concentrated liquid is obtained. 4. Filter the concentrate through a 400-mesh filter to obtain catnip extract. Example 2: Antioxidant test of nerve cells A. Materials and Instruments: 1. Cell line: Mouse brain neuroblastoma cell, purchased from ATCC (American Type Culture Collection), cell number CCL-131™, hereinafter referred to as Neuro2a cell. 2. Cell culture medium: DMEM (Dulbecco's Modified Eagle's medium) (purchased from Gibco, product number 12100-046), supplemented with 10% fetal bovine serum (purchased from Gibco, product number 10437-028), 1% antibiotic-antimycin (purchased from Thermo, product number 15240062) and 3.7 g / L sodium bicarbonate (purchased from Sigma, product number S8875-5006). 3. Trypsin: Prepared by diluting 10X trypsin (purchased from Gibco, product number 15400-054) with 9 times its volume of DPBS. 4. 35% H 2O 2. Solution: with H 2O It was prepared with 2 (purchased from Sigma, product number 1.08600) and PBS. 5. DCFH-DA reaction reagent: It was prepared with DCFH-DA (2',7'-dichlorodihydrofluorescein diacetate) (purchased from Sigma, product number SI-D6883-50MG) and DMSO (purchased from Sigma, product number 472301). 6. Flow cytometer, model BD Accuri, purchased from BD Corporation. B. Test Procedure: 1. Pack Neuro2a cells at a density of 2 × 10⁶ cells per well. 5 Neuro2a cells were seeded at a density of [number] cells / well in 6-well culture dishes containing 2 mL of cell culture medium per well and cultured at 37°C for 24 hours. The cells were then divided into three experimental groups: a blank control group, a control group, and an experimental group. Each group was performed in duplicate. 2. After 24 hours of incubation, each group was replaced with experimental culture medium. The blank and control groups used cell culture medium without samples; while the experimental groups used cell culture medium containing 0.25% (v / v) of the catnip extract prepared in Example 1. Each group was then incubated at 37°C for 1 hour. 3. After 1 hour of treatment, add DCFH-DA reagent to each group to make the final concentration 5 μg / mL, and treat each group at 37℃ for 15 minutes. 4. After 15 minutes of application, add 35% H2O to both the control and experimental groups. 2O The solution was prepared to a final concentration of 1 mM, and each group was incubated at 37°C for 1 hour. 5. Remove the experimental culture medium from each group after the reaction and rinse twice with PBS. 6. After rinsing, add 200 μL of trypsin to each well and react in the dark for 5 minutes. After the reaction, add cell culture medium to terminate the reaction. Then collect the suspended cells and cell culture medium from each well into the corresponding centrifuge tube, and centrifuge each tube to precipitate the cells. 7. Remove the supernatant from each centrifuge tube, wash the precipitated cells once with PBS, and then add 1 mL of PBS to each centrifuge tube to resuspend the cells to form a cell suspension. 8. After setting the excitation light parameters of the flow cytometer to 450-490 nm and the scattered light parameters to 510-550 nm, the green fluorescence signal of each group was detected by flow cytometry. C. Experimental Results: The relative ROS (reactive oxygen species) generation of all groups was calculated using the following formula: Relative ROS generation (%) = (green fluorescence signal of each group / green fluorescence signal of the blank group) × 100%. Statistically significant differences between the control group and other groups were determined using the Student's t-test. In the graph, "*" indicates a p-value less than 0.05 compared to the control group, "**" indicates a p-value less than 0.01 compared to the control group, and "***" indicates a p-value less than 0.001 compared to the control group. Please refer to Figure 1. Cells in the control group were not processed using samples and no H2 was added. 2O 2. Stimulation, therefore the experimental results of the blank group represent the cell performance under normal physiological metabolic conditions. Thus, with the relative ROS generation of the blank group set at 100%, the relative ROS generation of the control group was 145.1%, while the relative ROS generation of the experimental group was 51.9%. In other words, compared to the blank group, the cells in the control group showed significantly higher ROS generation after the addition of H+. 2O Following stimulation 2, the relative ROS production in the control group increased by approximately 45.1%. Compared to the control group, the experimental group cells, after the addition of catnip extract and H2O, showed a higher ROS production. 2O The experimental group showed a significant reduction in relative ROS production of approximately 64.2% after stimulation. Compared to the control group, the experimental group showed a reduction in relative ROS production of approximately 48.1%. Therefore, catnip extract can significantly reduce H 2O The amount of ROS generated in nerve cells due to 2. H 2O 2. It induces aerobic metabolism in cells, thus producing reactive oxygen species (ROS). Intracellular ROS attack macromolecules such as proteins, nucleic acids, and lipids, causing cell damage. In other words, experiments have shown that catnip extract can enhance the antioxidant capacity of nerve cells and reduce nerve cell damage. Catnip extract has the effect of enhancing antioxidant activity and reducing cell damage. Example three: nerve cells Gene test A. Materials and Instruments: 1. Cell line: Human neuroblastoma cells, purchased from ATCC, cell number CRL-2266™, hereinafter referred to as SHSY-5Y cells. 2. Cell culture medium: DMEM (Dulbecco's Modified Eagle's medium) (purchased from Gibco, product number 11965-092), supplemented with 10% fetal bovine serum (Fetal Bovine Serum) (purchased from Gibco, product number 10437-028) and 1% antibiotic-antimycin (purchased from Gibco, product number 15240-062). 3. RNA extraction reagent kit, purchased from TAN Bead, product number 301538. 4. SuperScript® III reverse transcriptase, purchased from Invitrogen, product number 18080-051. 5. ABI StepOnePlus TM ABI StepOnePlus Real-Time PCR System TM The Real-Time PCR system was purchased from Thermo Fisher Scientific. 6. KAPA SYBR FAST qPCR Master Mix (2X) Kit, purchased from Sigma, product number KM4102. B. Test Procedure: 1. Spread SHSY-5Y cells at a density of 1 × 10⁶ cells per well. 6 Cells were seeded at a density of [number] cells / well in 6-well culture dishes containing 2 mL of culture medium per well and cultured at 37°C for 24 hours. SHSY-5Y cells were then divided into two experimental groups: a control group and an experimental group. Each group was tested in triplicate. 2. After 24 hours of incubation, each group was replaced with experimental culture medium. The blank group received cell culture medium without the sample; and the experimental groups received cell culture medium containing 0.25% (v / v) of the catnip extract prepared in Example 1. Each group was then incubated at 37°C for 24 hours. 3. After culturing for 24 hours, SHSY-5Y cells from each group were collected. Then, RNA was extracted from the SHSY-5Y cells of each group using an RNA extraction reagent kit. 4. Take 1000 nanograms (ng) of extracted RNA from each group as a template, and use SuperScript® III reverse transcriptase to reverse transcribe the extracted RNA into the corresponding cDNA. 5. Using the ABI StepOnePlus™ real-time PCR system, quantitative real-time reverse transcription polymerase chain reaction (qPCR) was performed on cDNA from each group using the KAPA SYBR FAST qPCR Master Mix (2X) Kit and the primer combinations listed in Table 1. The expression levels of various target genes and their melting curves in the blank and experimental groups of SHSY-5Y cells were observed. The instrument settings for the qPCR were 95°C for 20 seconds, 95°C for 3 seconds, and 60°C for 30 seconds, repeated for 40 cycles. 6. Use 2 - ΔΔ Ct The method measures the relative expression level of the target gene. Relative expression level is defined as the fold change in RNA expression level of the target gene in the experimental or control group relative to the RNA expression level of the same gene in the control group. Therefore, by using cDNA for quantitative real-time reverse transcription polymerase chain reaction, the mRNA expression level of the gene can be indirectly quantified, thereby inferring the expression level of the protein encoded by the gene. 2 - ΔΔ Ct The method uses the cycling threshold of the GAPDH (glyceraldehyde 3-phosphate dehydrogenase) gene as the reference gene cycling threshold (Ct) for the internal control, and calculates the fold change according to the following formula: △Ct = Ct 實驗組之目標基因 / 空白組之目標基因 – Ct GAPDH △△Ct= △Ct 實驗組之目標基因 - △Ct 空白組之目標基因 Multiple change = 2 - ΔΔ Ct 平均值 Table 1 C. Experimental Results: The relative target gene expression level of all groups is calculated using the following formula: Relative target gene expression level = (Target gene expression level of each group / Target gene expression level of the blank group). The statistically significant difference between the experimental results of the control group and the experimental group was obtained by the Student's t-test. In the graph, "*" represents a p-value less than 0.05 compared with the control group, "**" represents a p-value less than 0.01 compared with the control group, and "***" represents a p-value less than 0.001 compared with the control group. Please refer to Figure 2. The cells in the control group were not treated with samples; therefore, the experimental results of the control group represent the cell performance under normal physiological metabolic conditions. Thus, in setting the relative... With a 1-fold increase in SIRT1 gene expression, the relative expression level in the experimental group was... The SIRT1 gene expression level was 9.40-fold. In other words, compared to the control group, the cells in the experimental group showed a 9.40-fold increase in expression after the addition of catnip extract. The expression level of the SIRT1 gene increased by approximately 840%. Therefore, it can be concluded that catnip extract can enhance nerve cells. The expression level of the SIRT1 gene. The SIRT1 gene is responsible for producing Sirt1 (Sirtuin 1), which has been shown to be associated with depression. Sirt1 expression in depressed patients is significantly lower than in healthy subjects. Sirt1 prevents depressive mood states by inhibiting excessive activation of glial cells in the brain and can also inhibit mood disorders caused by neuronal inflammation, thus improving mood function. In other words, experiments have demonstrated that catnip extract can reduce and / or inhibit excessive neuronal activation, reduce depression, and also reduce and / or inhibit neuronal inflammation, reduce mood disorders, and improve mood. Catnip extract has antidepressant and anti-depressant effects. Please refer to Figure 3. The cells in the control group were not treated with samples; therefore, the experimental results of the control group represent the performance of cells under normal physiological metabolic conditions. Thus, in setting the relative... TPH1 , DDC and When the expression level of the AANAT gene was 1-fold, the relative expression level in the experimental group was... The TPH1 gene expression level was 5.5-fold; the relative expression level in the experimental group was 5.5-fold. The expression level of the DDC gene was 13.2 times higher than that of the experimental group; while the relative expression level of the experimental group was 13.2 times higher. The AANAT gene expression level was 2.5-fold. In other words, compared to the control group, the cells in the experimental group showed a 2.5-fold increase in expression after the addition of catnip extract. The expression level of the TPH1 gene was significantly increased by approximately 450% after the addition of catnip extract to the cells in the experimental group, compared to the control group. The expression level of the DDC gene was significantly increased by approximately 1220%. Compared to the control group, the cells in the experimental group, after being supplemented with catnip extract, showed a relative increase in expression. The expression level of the AANAT gene increased by approximately 150%. Therefore, it can be concluded that catnip extract can enhance nerve cells. TPH1 and The expression level of the DDC gene. The TPH1 gene is responsible for producing tryptophan hydroxylase 1 (TPH1). TPH1 is responsible for synthesizing serotonin. And... The DDC gene is responsible for producing enzymes involved in the production of dopamine and serotonin. In other words, experiments have shown that catnip extract can enhance the synthesis of serotonin in nerve cells. Since serotonin can keep people alert and happy, catnip extract also has the effect of keeping people alert and happy. Therefore, it can be concluded that catnip extract can enhance nerve cells. Expression level of the AANAT gene. The AANAT gene is a key regulator of the human circadian rhythm, responsible for producing an enzyme involved in melatonin synthesis. This enzyme is crucial for catalyzing the conversion of serotonin into melatonin. In other words, experiments have demonstrated that catnip extract can enhance the conversion of serotonin into melatonin by nerve cells, thereby increasing the synthesis of melatonin. Catnip extract thus increases melatonin levels. Since melatonin promotes restful sleep and regulates the biological clock, catnip extract can help with nighttime sleep, promoting restful sleep and regulating the biological clock. Example 4: Human Experimentation A. Experimental Procedure: Seven adult participants aged 20 to 55 with poor mental health or sleep problems were instructed to consume one bottle of the experimental beverage daily for four weeks (28 days). The experimental beverage contained 3 g of catnip extract prepared in Example 1 and 47 g of water. Sleep monitoring, a somatosensory questionnaire, and blood tests were conducted on the participants before starting the beverage (Week 0), 14 days after starting the beverage (Week 2), and 28 days after starting the beverage (Week 4). Sleep monitoring involved using an electrocardiogram (ECG) to measure the percentage of light sleep, deep sleep, and sleep onset time before and after medication administration. The ECG system is based on the CPC cardiopulmonary coupling theory developed by Harvard University, analyzing sleep patterns through heart rate to detect the subject's states of deep sleep, light sleep, REM sleep, and wakefulness per minute, accurately monitoring sleep data. In human trials, subjects wore an ECG device manufactured by Dali Cloud Health Technology Co., Ltd. for the monitoring. The somatosensory questionnaire survey used a sleep status questionnaire to allow participants to self-assess before and after taking the medication. The sleep status questionnaire is shown in Table 2 below. Participants self-assessed their sleep quality and daytime activity levels. Each participant self-assessed their sleep status, judging these sleep conditions and selecting one of five options: very good (5 points), quite good (4 points), average (3 points), poor (2 points), and very poor (1 point). Table 2 The blood test involves drawing blood from the subject on an empty stomach, and then having the subject's serum sample sent to Li-Jen Medical Laboratory (Taiwan) to measure the changes in serotonin levels in the subject's blood before and after taking the medication. B. Experimental Results: Statistically significant differences between week 0 and other weeks, and between week 2 and other weeks, were obtained using Student's t-test statistical analysis. In the graph, "*" indicates a p-value less than 0.05 compared to week 0, "**" indicates a p-value less than 0.01 compared to week 0, and "***" indicates a p-value less than 0.001 compared to week 0. Similarly, "#" indicates a p-value less than 0.05 compared to week 2, "##" indicates a p-value less than 0.01 compared to week 2, and "###" indicates a p-value less than 0.001 compared to week 2. Please refer to Figure 4. Figure 4 shows the sleep onset time of one subject before and after consuming the experimental beverage. The subject's sleep onset time was 21 minutes at week 0, while at week 4 (i.e., after 4 weeks of continuous consumption of catnip extract), the sleep onset time was 8 minutes. In other words, compared to before consumption, continuous consumption of catnip extract for 4 weeks shortened this subject's sleep onset time by 13 minutes, or 61.9%. This demonstrates that catnip extract can indeed shorten sleep onset time. In other words, catnip extract has the effect of helping to induce sleep quickly. Please refer to Figure 5. Figure 5 shows the average relative light sleep percentage of the 7 subjects before and after consuming the experimental beverage. The light sleep percentage measured before consumption by the 7 subjects is considered as 100% relative light sleep percentage. At this point, the relative light sleep percentage at week 4 (i.e., after 4 weeks of continuous consumption of catnip extract) is 96.5%. In other words, compared to before consumption, continuous consumption of catnip extract for 4 weeks reduced the relative light sleep percentage of these subjects by 3.5%. This shows that catnip extract can indeed shorten the light sleep percentage. Please refer to Figure 6. Figure 6 shows the average relative deep sleep percentage of the 7 subjects before and after consuming the experimental beverage. The deep sleep percentage measured before consumption by the 7 subjects is considered as 100% relative deep sleep percentage. At this point, the relative deep sleep percentage in week 4 (i.e., after 4 weeks of continuous consumption of catnip extract) is 112.9%. In other words, compared to before consumption, continuous consumption of catnip extract for 4 weeks increased the relative deep sleep percentage of these subjects by 12.9%. This shows that catnip extract can indeed increase the percentage of deep sleep. In other words, catnip extract has the effect of increasing deep sleep. Since the brain and body are in a state of complete rest during deep sleep, which is the main sleep for eliminating physical fatigue, catnip extract has the effect of allowing the body to rest completely and relieving fatigue. Please refer to Figure 7. Figure 7 shows the average sleep quality scores of 7 subjects before and after consuming the experimental beverage. The sleep quality score was 2.7 at week 0, 3.4 at week 2 (i.e., after 2 weeks of continuous consumption of catnip extract), and 3.4 at week 4 (i.e., after 4 weeks of continuous consumption of catnip extract). In other words, compared to before consumption, continuous consumption of catnip extract for 2 weeks significantly increased the sleep quality scores of these subjects by 25.9%, and continuous consumption of catnip extract for 4 weeks significantly increased their sleep quality scores by 25.9%. Furthermore, the proportion of subjects showing improvement was 57.1% (4 subjects) at week 2, and 85.7% (6 subjects) at week 4. This demonstrates that catnip extract can indeed improve sleep quality. Please refer to Figure 8. Figure 8 shows the average daytime activity energy scores of 7 subjects before and after consuming the experimental beverage. The daytime activity energy score was 3.2 at week 0, 3.6 at week 2 (i.e., after 2 weeks of continuous consumption of catnip extract), and 3.9 at week 4 (i.e., after 4 weeks of continuous consumption of catnip extract). In other words, compared to before consumption, continuous consumption of catnip extract for 2 weeks increased the daytime activity energy score of these subjects by 12.5%, and continuous consumption of catnip extract for 4 weeks increased the daytime activity energy score by 21.9%. Furthermore, the improvement rate was 85.7% (6 subjects). This shows that catnip extract can indeed improve daytime activity energy. In other words, catnip extract has the effect of improving daytime energy and daytime mental state. Please refer to Figure 9. Figure 9 shows the average serotonin levels of the 7 subjects before and after consuming the experimental beverage. The serotonin level was approximately 74.0 ng / mL at week 0, approximately 84.4 ng / mL at week 2 (i.e., after 2 weeks of continuous catnip extract consumption), and approximately 92.9 ng / mL at week 4 (i.e., after 4 weeks of continuous catnip extract consumption). In other words, compared to before consumption, continuous consumption of catnip extract for 2 weeks significantly increased the subject's serotonin level by 14.1%, and continuous consumption for 4 weeks significantly increased it by 25.5%. Compared to after 2 weeks, continuous consumption of catnip extract for 4 weeks significantly increased the subject's serotonin level by 10.1%. Furthermore, the proportion of subjects showing improvement was 85.7% (6 subjects) at week 2, and 100% (7 subjects) at week 4. This demonstrates that catnip extract can indeed increase serotonin levels. Serotonin is one of the raw materials for melatonin. A deficiency in serotonin can easily lead to insomnia or sleep disorders. In other words, catnip extract can indeed improve sleep. Example 5 : Compound analysis test A. Materials and Instruments: 1. Nuclear Magnetic Resonance Spectrometer (NMR): 1D and 2D spectra were obtained using an Ascend 400 MHz spectrometer purchased from Bruker Co., Germany. Chemical shift is expressed as δ in ppm. 2. High-resolution liquid chromatography-mass spectrometry: Measured using a tandem ultra-high performance liquid chromatography system (Ultimate 3000 HPLC, purchased from Thermo Fisher Scientific) and a high-resolution orbital ion trap mass spectrometer (Q-EXACTIVE System with Ion Max Source, purchased from Thermo Fisher Scientific), with units in m / z. 3. Medium pressure liquid chromatography (MPLC): CombiFlash® Rf+, purchased from Teledyne ISCO, Lincoln, Nebraska, USA. 4. High Performance Liquid Chromatography (HPLC): Hitachi chromaster 5260 series, purchased from Hitachi, Tokyo, Japan; extraction solvent delivery system: Hitachi chromaster 5110; column temperature control system: Hitachi chromaster 5310; diode array detector (DAD): Hitachi chromaster 5430, detection wavelength: 250 nm. 5. HPLC analysis column: Mightysil RP-18 GP 250 (250 x 4.6 mm, 5 μm, purchased from Kanto, Tokyo, Japan). 6. HPLC separation column: Luna® 5μm C18(2) 100 Å (250 x 10 mm), purchased from Phenomenex, USA. 7. Column Chromatography Packing Materials: (1) Macroporous resin: Diaion HP-20, purchased from Mitsubishi Chemical Corporation, Japan. (2) Normal phase silicone: Merck Kieselgel 60, 40-63 um, purchased from Merck, Germany, product number Art. 9385. (3) Reverse phase silicone: Merck LiChroprep® RP-18, 40-63 um, purchased from Merck, Germany, product number Art. 0250. 8. Thin-Layer Chromatography: (1) TLC aluminum sheet: Thin-layer chromatography sheet, coated with silicone 60F 254 (0.25 mm), purchased from Merck, Germany. (2) TLC aluminum sheet: Thin-layer chromatography sheet, coated with RP-18 F 254-S (0.25 mm), purchased from Merck, Germany. 9. Ultraviolet lamp (UV Lamp): UVP UVGL-25, with wavelengths of 254 nm and 365 nm. 10. Solvents: methanol, acetonitrile, n-butanol, chloroform-d1 (99.5% deuteration), methanol-d4 (99.5% deuteration), heavy water (deuterium oxide, >99.8% deuteration), and dimethyl monoxide-d6 (>99.9% deuteration), all purchased from Merck, Taiwan. 11. Ultrapure water (18.2 M): from Millipore Synergy® water purification system (purchased from Millipore, USA). 12. Chemical reagents: Methanol (HPLC grade) and formic acid (ACS grade), purchased from Merck, Taiwan. 13. Filter membrane: Polyvinylidene fluoride membrane filters (PVDF), pore size 0.22 microns, purchased from Millipore, USA. 14. Sample: Catnip extract prepared in Example 1. B. HPLC analysis procedure: 1. Filter the catnip extract using a filter membrane. 2. The catnip extract was analyzed using HPLC. Methanol (A) and water (B) were used as solvents, with the addition of 0.1% formic acid. The flow rate was set to 1 mL / min, and the injection volume of catnip extract was 10 μL. Extraction conditions: at 0 min, methanol (A):water (B) = 2:98; at 10 min, A:B = 2:98; at 40 min, A:B = 70:30; at 50 min, A:B = 100:0; at 60 min, A:B = 100:0. The column temperature was 40 °C during HPLC analysis. 3. Please refer to Figure 10. The HPLC fingerprint of the catnip extract has many distinct peaks (including peaks 01-07, 09-14, and 16-17). C. Compound isolation and structural identification process: 1. In order to separate the fat-soluble and water-soluble components of catnip extract, 10 L of catnip extract was subjected to liquid-liquid phase partition extraction using n-butanol as solvent to obtain 42.3 g of n-butanol-soluble fraction (fat-soluble component) and 131.3 g of water-soluble fraction (water-soluble component). 2. The separation and purification of the functional compounds were carried out using a bioassay-guided fractionation method. 3. The 42.3 g of the n-butanol-soluble fraction was further separated by column chromatography on a Diaion HP-20 macroporous resin column. Using pure water, a 1:1 (v / v) mixture of pure water and methanol, and methanol as eluents in sequence, three separated fractions (hereinafter referred to as the BUF1 fraction, the BUF2 fraction, and the BUF3 fraction) were obtained. 4. The BUF-3 fraction was taken and re-separated by reverse-phase medium-pressure liquid chromatography (RP-MPLC) to obtain various eluates. Here, the eluent was linearly eluted from water to methanol for 120 minutes at a flow rate of 10 mL per minute. Subsequently, thin-layer chromatography (TLC aluminum sheet: TLC plate, coated with RP-18 F 254-S (0.25 mm)) was used to combine the eluates with similar results, and five sub-separated fractions (hereinafter referred to as the BUF3-1 fraction, the BUF3-2 fraction, the BUF3-3 fraction, the BUF3-4 fraction, and the BUF3-5 fraction) were obtained. 5. The BUF3-3 fraction was purified by reverse-phase HPLC (methanol / water = 11 / 9) to obtain compound TCI-CatM-03. After analyzing its chemical structure by hydrogen nuclear magnetic resonance spectroscopy (1H-NMR) and carbon-13 nuclear magnetic resonance spectroscopy (13C-NMR), and comparing the data with Reference 1 (Flavonoid and phenolic glycosides from Salvia offcinalis. Yinrong Lu, L. Yeap Foo. Phytochemistry, 2000, 55, 263-267.), it was confirmed that compound TCI-CatM-... 6. BUF3-2 fraction was purified by reverse HPLC (methanol / water = 1 / 1) to obtain compound TCI-CatM-16. Its chemical structure was analyzed by 1H-NMR and 13C-NMR spectroscopy, and compared with that in Reference 2 (Synthesis and Characterization of Bimetallic Nanoclusters Stabilized by Chiral and Achiral Polyvinylpyrrolidinones. Catalytic C(sp...). 3 (–H Oxidation. Huafang Fan, Zongbo Tong, Zhaoyang Ren, Kanchan Mishra, Shunya Morita, Edruce Edouarzin, Lingaraju Gorla, Boris Averkiev, Victor W. Day, Duy H. Hua. J. Org. Chem. 2022, 87, 10, 6742–6759.) Comparison data confirmed that compound TCI-CatM-16 is 7a-hydroxy-3,6-dimethyl-2,4,5,6,7,7a-hexahydrocyclohexa[1,2-b]furan-2-one, as shown in Figures 13 and 14. The names and chemical structural formulas of the aforementioned compounds are shown in Table 3 below. Table 3 In summary, the use of catnip extract in improving sleep and antidepressant effects in any embodiment relates to the use of catnip extract in preparing a composition for improving sleep or antidepressant effects, thereby providing a composition that, when applied to an individual, produces an effect of improving sleep or antidepressant effects in that individual. In other words, the aforementioned composition has the function of improving sleep or antidepressant effects. In some embodiments, the composition obtained from catnip extract also has one or more of the following functions: shortening sleep onset time and / or improving sleep quality, increasing daytime activity energy, antidepressant effects, and brain protection. none Figure 1 is a bar chart of the cell experiment results relative to ROS production. Figure 2 is a relative... Figure 3 is a bar chart showing the cell experiment results of SIRT1 gene expression levels. Figure 4 is a bar chart showing the human experiment results of sleep onset time in weeks 0 and 4. Figure 5 is a bar chart showing the human experiment results of the relative percentage of light sleep in weeks 0 and 4. Figure 6 is a bar chart showing the human experiment results of the relative percentage of deep sleep in weeks 0 and 4. Figure 7 is a bar chart showing the human experiment results of the relative sleep quality in weeks 0, 2, and 4. Figure 8 is a bar chart showing the human experiment results of the relative daytime activity level in weeks 0, 2, and 4. Figure 9 is a bar chart showing the human experiment results of serotonin levels in weeks 0, 2, and 4. Figure 10 is the HPLC fingerprint of catnip extract. Figure 11 is the hydrogen-NMR spectrum of compound I. Figure 12 is the carbon-13 NMR spectrum of compound I. Figure 13 is the hydrogen-NMR spectrum of compound II. Figure 14 is the carbon-13-NMR spectrum of compound II. TW202432165A_112151732_SEQL.xml

Claims

1. Use of a catnip extract for preparing a composition that enhances daytime activity, wherein the catnip extract is obtained by extracting catnip spikes with water at 85±5°C.

2. The use as described in claim 1, wherein the catnip extract has the effect of increasing the expression of the synthetic serotonin gene.

3. The use as described in claim 2, wherein the catnip extract has the effect of increasing the expression levels of the TPH1 and / or DDC genes in nerve cells.

4. The use as described in claim 1, wherein the catnip extract has the effect of increasing the expression of the melatonin gene.

5. Use of a catnip extract for preparing a composition that increases serotonin levels in the blood, wherein the catnip extract is obtained by extracting catnip spikes with water at 85±5°C.

6. The use as claimed in claim 5, wherein the catnip extract has the effect of increasing the expression levels of the TPH1 and / or DDC genes in nerve cells.

7. A catnip extract having the use as described in claim 1 or 5, comprising one or a combination of compounds of formula I to formula II.

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