Application of wood frog seeds and extracts thereof in preparation of products for preventing and improving depression
By preparing water extracts and oil components from frog seeds and combining them with an acceptable carrier to form a composition, the problem of significant side effects of existing drugs has been solved. This approach effectively alleviates depression caused by chronic stress, increases neurotransmitter levels, and reduces inflammatory factor levels, providing a new intervention strategy for depression.
Patent Information
- Application Number
- CN202511351171.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-22
- Publication Date
- 2025-11-21
AI Technical Summary
Existing chemically synthesized drugs have significant side effects when used to treat depression, resulting in poor patient compliance. Furthermore, the functional research and development of high-value-added products from forest frog seeds have not been fully utilized, especially in terms of antidepressant effects, where no literature reports have been found.
Using frog seeds and their extracts, water extracts and oil components of frog seeds are prepared by hydrolysis and supercritical carbon dioxide extraction. These components are then combined with pharmaceutically or food-acceptable carriers to form a composition for the preparation of products that prevent and improve depression.
Forest frog seed extract significantly improves depression caused by chronic stress, increases the level of neurotransmitter 5-HT, reduces the levels of inflammatory factors TNF-α and IL-1β, alleviates depressive behavior, and provides a safe and natural intervention for depression.
Smart Images
Figure CN120983480A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of cognitive health, and particularly relates to application of rana chensinensis seeds and extracts thereof in preparation of products for preventing and improving depression, and is especially suitable for preparation of products for preventing and improving depression caused by chronic stress. BACKGROUND
[0002] With the increasingly fierce social competition and the increasing life pressure, depression disorder has become a global major public health problem. According to the report of the World Health Organization, more than 350 million people of different ages worldwide are deeply troubled by depression, and the incidence rate shows a rising trend year by year. In addition to the patients with depression reaching the clinical diagnosis standard, a large number of people are in the sub-health state of emotional depression, interest decline and energy deficiency due to long-term chronic stress, emotional stimulation and other factors, i.e. "depression state". At present, the main treatment for depression in the clinic is mainly chemical synthetic drugs. Although such drugs have certain efficacy, they are often accompanied by obvious side effects such as nausea, vomiting, insomnia, sexual dysfunction and weight gain, resulting in poor patient compliance.
[0003] Rana chensinensis seeds are traditionally mainly regarded as a food material in the northeast of China. However, the utilization of rana chensinensis seeds is currently limited to rough processing and traditional eating methods, and the research on deep functional components and the development of high value-added products are still insufficient. Existing researches are mainly focused on the chemical components, immune regulation and skin care effects of rana chensinensis oil, while the systematic research on rana chensinensis seeds, especially the specific physiological functions, is still in the preliminary stage. In the field of mental health, the potential application of rana chensinensis seeds and extracts thereof in the aspect of anti-depression has not been reported in the literature. SUMMARY
[0004] The application provides application of rana chensinensis seeds and extracts thereof in preparation of products for preventing and improving depression, which is a safe and effective product for preventing and improving depression state and is of natural origin.
[0005] The technical scheme adopted by the application is that rana chensinensis seeds and extracts thereof are applied in preparation of products for preventing and improving depression.
[0006] The product according to the application is a drug, a health food, a special diet food or a special medical purpose food.
[0007] The depression according to the application is depression caused by chronic stress.
[0008] The application improves despair, loss, reaction retardation behavior, reduction of neurotransmitter 5-HT level or increase of inflammatory factors TNF-α and IL-1β level caused by chronic stress.
[0009] The rana chensinensis seeds according to the application are fresh rana chensinensis seeds, dried rana chensinensis seeds or freeze-dried powder.
[0010] The forest frog seed extract of the present application is a mixture extracted from the hydrolyzed forest frog seed or an oil component extracted from the forest frog seed.
[0011] The mixture extracted from the hydrolyzed forest frog seed of the present application is obtained by the following steps: a) mixing the forest frog seed raw material with water at a mass ratio of 1:10 to 1:20, homogenizing to obtain a forest frog seed slurry; b) adding a complex protease to the slurry obtained in step a) for hydrolysis, the complex protease comprising trypsin, papain and flavor protease, the addition amount being 2.0%-5.0% of the weight of the forest frog seed raw material, stirring under suitable temperature and pH conditions; c) inactivating the enzyme by heat treating the reaction solution after enzymolysis in step b) at 105°C for 5-10 minutes, and then water extracting at 60-80°C for 1-2 hours; d) centrifuging or filtering the mixture obtained in step c) to collect the clarified liquid; e) vacuum concentrating the clarified liquid obtained in step d) and then spray drying to obtain the mixture.
[0012] The oil component extracted from the forest frog seed of the present application is obtained by the following steps: (1) washing, drying and crushing the forest frog seed; (2) placing the forest frog seed raw material treated in step (1) in an extraction kettle, using supercritical carbon dioxide as the extraction solvent, and dynamically extracting under the conditions of a pressure of 30-40 MPa and a temperature of 45-55°C for 3-5 hours; (3) introducing the supercritical carbon dioxide fluid carrying the forest frog seed oil into a separation kettle, releasing the pressure to recover the forest frog seed oil.
[0013] A composition for preventing and improving depression, the active ingredient of which comprises forest frog seed and its extract, and one or more pharmaceutically or food-acceptable carriers, diluents or excipients.
[0014] The weight percentage of the forest frog seed water extract in the composition of the present application is 5-45%, or the volume percentage of the forest frog seed oil is 10-60%.
[0015] The daily administration dose of the forest frog seed water extract in the composition of the present application is 1-10 grams, and the daily administration dose of the forest frog seed oil is 0.5-4 grams.
[0016] The present application has the following advantages: The application discloses the water extract of Ranae Semen and the oil of Ranae Semen for the first time in the prevention and improvement of depressive state, especially the depression caused by chronic stress model, which provides a new solution for the intervention of depression. The water extract of Ranae Semen and the oil of Ranae Semen not only improve the depressive behavior state, but also play a role in neurotransmitters (improve 5-HT level) and immune inflammatory factors (reduce TNF-α and IL-1β level). BRIEF DESCRIPTION OF DRAWINGS
[0017] Figure 1 is a result graph of the open field experiment; Figure 2 is a result graph of the tail suspension experiment; Figure 3 is a result graph of the elevated plus maze experiment; Figure 4 is a graph of the 5-HT level of hippocampal tissue; Figure 5 is a graph of the TNF-α level in serum; Figure 6 is a graph of the IL-1β level in serum. DETAILED DESCRIPTION
[0018] Example 1 Water extract of Ranae Semen after hydrolysis a) 1.0 kg of dried Ranae Semen is mixed with 15 kg of purified water (the ratio of material to liquid is 1:15) to homogenize; b) the slurry is transferred to a controllable temperature reaction kettle, the temperature and pH of the system are adjusted (50 DEG C, pH 7.0), then 30 g of compound protease is added (the addition amount is 3.0% of the weight of the Ranae Semen raw material), the three enzymes are compounded according to the mass ratio of 1:1:1, constant temperature stirring is carried out for 4 hours; c) the reaction liquid is heated to 105 DEG C and maintained for 8 minutes to inactivate the enzyme, then the temperature is reduced to 70 DEG C, and constant temperature stirring extraction is continued for 1.5 hours; d) the mixture after extraction is centrifuged at 8000 rpm, and the supernatant is collected; e) after concentration, spray drying is carried out (the inlet air temperature is 180 DEG C, and the outlet air temperature is 85 DEG C), to obtain about 215 g of light yellow Ranae Semen hydrolysis extract powder, and the yield is 21.5%.
[0019] Example 2 Water extract of Ranae Semen after hydrolysis a) 1.0 kg of dried Ranae Semen is mixed with 10 kg of purified water (the ratio of material to liquid is 1:10) to homogenize; b) the slurry is transferred to a controllable temperature reaction kettle, the temperature and pH of the system are adjusted (50 DEG C, pH 7.0), then 20 g of compound protease is added (the addition amount is 2.0% of the weight of the Ranae Semen raw material), the three enzymes are compounded according to the mass ratio of 1:1:1, constant temperature stirring is carried out for 3 hours; c) The reaction solution was heated to 105°C for 5 minutes to inactivate the enzyme, and then the temperature was lowered to 60°C and the extraction was continued for 1 hour at constant temperature; d) The mixture after extraction was centrifuged at 8000 rpm, and the supernatant was collected; e) After concentration, spray drying was performed (inlet temperature 180°C, outlet temperature 85°C) to obtain a light yellow powder of the hydrolyzed extract of Ranae semen.
[0020] Example 3: Extracting mixture from Ranae semen after hydrolysis a) 1.0 kg of dried Ranae semen was mixed with 20 kg of purified water (solid-liquid ratio 1:20) and homogenized. b) The slurry was transferred to a temperature-controllable reaction kettle, and the temperature and pH of the system were adjusted (50°C, pH 7.0), and then 50 g of compound protease was added (the addition amount was 5.0% of the weight of the Ranae semen raw material), and the three enzymes were compounded in a mass ratio of 1:1:1, and constant temperature stirring was carried out for 5 hours. c) The reaction solution was heated to 105°C for 10 minutes to inactivate the enzyme, and then the temperature was lowered to 80°C and the extraction was continued for 2 hours at constant temperature; d) The mixture after extraction was centrifuged at 8000 rpm, and the supernatant was collected; e) After concentration, spray drying was performed (inlet temperature 180°C, outlet temperature 85°C) to obtain a light yellow powder of the hydrolyzed extract of Ranae semen.
[0021] Example 4: Extracting oil components from Ranae semen (1) 1.0 kg of dried Ranae semen was crushed with a crusher and passed through a 40-mesh sieve to obtain Ranae semen powder. (2) The above Ranae semen powder was evenly loaded into an extraction kettle, the extraction pressure was 35 MPa, the extraction temperature was 50°C, the CO2 flow rate was 80 L / h, and dynamic extraction was carried out for 4 hours. (3) The Ranae semen oil obtained was collected from the outlet valve at the bottom of the separation kettle every hour, and after the extraction was completed, the weight was measured, and a total of 136 g of light yellow transparent Ranae semen oil was obtained, with a yield of 13.6%.
[0022] Example 5: Extracting oil components from Ranae semen (1) 1.0 kg of dried Ranae semen was crushed with a crusher and passed through a 40-mesh sieve to obtain Ranae semen powder.
[0023] (2) The above Ranae semen powder was evenly loaded into an extraction kettle, the extraction pressure was 40 MPa, the extraction temperature was 55°C, the CO2 flow rate was 80 L / h, and dynamic extraction was carried out for 5 hours.
[0024] (3) From the outlet valve at the bottom of the separation kettle, the obtained oil of forest frog seed was collected once every certain period of time, and weighed after the extraction was completed. A total of 145 g of transparent yellow oil of forest frog seed was obtained, with a yield of 14.5%.
[0025] Example 6: Extraction of oil components from forest frog seed (1) 1.0 kg of dried forest frog seed was taken, crushed by a pulverizer, and passed through a 40-mesh sieve to obtain forest frog seed powder.
[0026] (2) The above forest frog seed powder was evenly loaded into an extraction kettle, and dynamic extraction was performed at an extraction pressure of 30 MPa, an extraction temperature of 45°C, and a CO2 flow rate of 80 L / h for 3 hours.
[0027] (3) From the outlet valve at the bottom of the separation kettle, the obtained oil of forest frog seed was collected once every certain period of time, and weighed after the extraction was completed. A total of 113 g of transparent yellow oil of forest frog seed was obtained, with a yield of 11.3%. Example 7
[0028] Fifty 4-6-week-old male C57BL / 6 mice were randomly divided into five groups: a blank control group (CG), a model group (MD), a forest frog seed group (LWZ), a forest frog seed water extract group (LWZT), and a forest frog seed oil group (LWZY), with 10 mice in each group. The CG and MD groups were given the same amount of normal saline by gavage and fed with normal feed, the LWZ group was fed with feed containing 30wt% forest frog seed, the LWZT group was given forest frog seed water extract 400 mg / kg by gavage, and the LWZY group was given forest frog seed oil 400 mg / kg by gavage, for 28 consecutive days. Then, the MD group, the LWZ group, the LWZT group, and the LWZY group were subjected to chronic stress to establish a depression model, and the mice continued to be given gavage during the modeling period. The mice were stimulated by noise for 30 min, inclined mouse cage for 24 h, cold water bath for 5 min, suspension for 30 min, continuous light for 24 h, fasting or water deprivation for 24 h, etc. One to two kinds of stimulation were given to the mice every day, and the stimulation lasted for 45 days. The open field test was performed 30 minutes after the last administration. The mice were placed in the center of the open field box to freely explore for 10 minutes, and the motion trail was recorded using an animal behavior analysis system to calculate the central area residence time. The results showed that the central area residence time of the LWZ, LWZT, and LWZY groups was significantly higher than that of the MD group, indicating that forest frog seed and its extracts can alleviate the depression-like behavior of mice (see Figure 1 ). Example 8
[0029] The tail suspension test was performed 24 hours after the open field test. The mouse tail was fixed to the support with adhesive tape, and the mouse was hung upside down. The cumulative immobility time of the mouse in the last 4 minutes was recorded for 6 minutes. The results showed that, compared with the model group, the forest frog seed water extract and the forest frog seed oil group can reduce the immobility time of the last 4 minutes of the depressed mice. Figure 2The MD group showed a significantly longer immobility time of 4 minutes compared to the CG group, suggesting that the MD group mice may exhibit depressive-like behavior. The LWZ, LWZT, and LWZY groups showed significantly shorter immobility times than the MD group, indicating that *Rhizophora stylosa* seeds and their extracts have a mitigating effect on depressive-like behavior in mice (see...). Figure 2 ). Example 9
[0030] Twenty-four hours after the tail suspension test, an elevated cross maze test was performed. Mice were gently placed in the central area of the maze and allowed to explore freely for 5 minutes. The number of times the mice entered the open and closed arms and the duration of their stay were recorded. The results showed that, compared with the model group, the water extract and oil groups of *Rhizophora stylosa* seeds increased the exploration time of mice in the open arms. The results showed that the open arm stay time of the LWZ, LWZT, and LWZY groups was significantly higher than that of the MD group, indicating that *Rhizophora stylosa* seeds and their extracts can alleviate the depressive state of mice (see...). Figure 3 ). Example 10
[0031] Mice were euthanized after the above experiments, and hippocampal tissue was collected for 5-HT testing. Blood was collected from the eyeballs and placed in a 4°C freezer for 30 min. The supernatant was then centrifuged at 8000 r / min, 4°C for 20 min, and the supernatant was used to detect TNF-α and IL-1β levels. The results showed (see...) Figures 4-6 The frog seeds and their extracts significantly increased the 5-HT level in the hippocampus of depressed mice and decreased the serum levels of TNF-α and IL-1β.
[0032] Example 11 Preparation of soft capsules Prepare soft capsules using 0.25g of frog seed oil, 0.2g of corn oil, 0.05g of DHA algal oil, 5mg of vitamin E, gelatin, glycerin, and purified water. The recommended daily dose is 2 capsules for daily relief of depressive moods.
[0033] Example 12 Preparation of oral liquid Prepare an oral liquid with 10g of frog seed water extract powder, 5g of Schisandra chinensis extract, 5g of Angelica sinensis extract, 50g of fructooligosaccharides, 0.5g of potassium sorbate, and purified water. It is recommended to take one vial (10mL) twice a day to help improve depressive symptoms.
[0034] Example 13 Preparation of Powder Ingredients: Frog seed water extract powder (15g), Centella asiatica extract (15g), dried tangerine peel powder (15g), maltodextrin (10g), sour cherry powder (5g), steviol glycosides (0.1g), and edible flavoring (strawberry flavor, 0.2g). Take one sachet daily to relieve low mood, fatigue, and lethargy caused by chronic exercise stress, and help maintain a good mental state. Example 14
[0035] For stress-induced depression, Linna frog seed oil soft capsules can be taken, and it is recommended to take 2 capsules per day, 1 capsule each time, for 8 weeks, to help alleviate the depression caused by stress.
[0036] Example 15 Preparation of chewable tablets Linna frog seed water extract powder 800g, mint powder 100g, dried tangerine peel powder 100g, sorbitol 150g, magnesium stearate 20g, gelatinized starch 100g, and a suitable amount of peppermint essence are used to prepare chewable tablets, and it is recommended to take 4-6 tablets per day to help alleviate mild stress and improve low mood. Example 16
[0037] Linna frog seed oil 500mg, DHA algal oil 100mg, soybean oil 200mg, vitamin E 20mg, gelatin, glycerol, purified water, and titanium dioxide are used to prepare soft capsules according to the conventional method, and the net content of each oval soft capsule is 520mg.
[0038] Example 17 Preparation of solid beverage Linna frog seed water extract powder (40g), soybean protein powder (30g), whey protein powder (10g), sour cherry powder (10g), malt dextrin (15g), steviol glycoside (0.1g), and strawberry powder (0.2g) are used to prepare a solid beverage of 25g per bag, which can be used to improve post-exercise depression.
[0039] Example 18 Preparation of oral liquid Linna frog seed water extract liquid (50g by nitrogen content), lily extract (10g), high-quality ginger extract (10g), sour jujube extract (10g), isomaltooligosaccharide (50g), potassium sorbate (0.5g), and purified water (added to 1000mL) are used to prepare an oral liquid. The Linna frog seed water extract liquid, sour jujube and lily extracts are dissolved in purified water, and isomaltooligosaccharide is added while stirring. After dissolution, preservatives are added, and the volume is adjusted. The mixture is filtered and filled into 30mL oral liquid bottles, and then sterilized to obtain the product. It is used to improve daily depression. Example 19
[0040] Linna frog seed oil (8g), soybean protein (8g), pectin (1g), carrageenan (1g), malt dextrin (2g), dietary fiber (2g), vitamin and mineral complex (1.5g), emulsifier, stabilizer, and purified water (appropriate amount) are used to prepare an oil-in-water emulsion by high-speed shearing and homogenization process. After sterilization, it is aseptically filled. It can be used to improve daily depression.
Claims
1. Application of frog seeds and their extracts in the preparation of products for preventing and improving depression.
2. The application according to claim 1, characterized in that: The products mentioned are pharmaceuticals, health foods, special dietary foods, or foods for special medical purposes.
3. The application according to claim 1, characterized in that: The depression mentioned refers to depression caused by chronic stress.
4. The application according to claim 3, characterized in that: It can improve despair, loss, slowed-response-like behavior, decreased levels of the neurotransmitter 5-HT, or increased levels of inflammatory factors TNF-α and IL-1β caused by chronic stress.
5. The application according to claim 1, characterized in that: The frog eggs mentioned are fresh frog eggs, dried frog eggs, or freeze-dried powder.
6. The application according to claim 1, characterized in that: The frog seed extract is a mixture extracted from frog seeds after hydrolysis or an oil component extracted from frog seeds.
7. The application according to claim 6, characterized in that: The mixture extracted from the frog eggs after hydrolysis is obtained through the following steps: a) Mix the raw material of frog eggs with water at a mass ratio of 1:10 to 1:20, homogenize the mixture, and obtain frog egg slurry. b) Add a complex protease to the slurry obtained in step a) for hydrolysis. The complex protease includes trypsin, papain and flavor protease. The amount added is 2.0%-5.0% of the weight of the frog seed raw material. Stir the reaction under suitable temperature and pH conditions. c) Heat the reaction solution after enzymatic hydrolysis in step b) at 105℃ for 5-10 minutes to inactivate the enzyme, and then extract it with water at 60-80℃ for 1-2 hours. d) Centrifuge or filter the mixture obtained in step c) and collect the clear liquid; e) The clarified liquid obtained in step d) is concentrated under vacuum and then spray-dried to obtain the final product.
8. The application according to claim 6, characterized in that: The oil components extracted from the frog eggs were obtained through the following steps: (1) Wash, dry and crush the frog eggs; (2) Place the frog eggs processed in step (1) into an extraction vessel and use supercritical carbon dioxide as the extraction solvent to dynamically extract for 3-5 hours under a pressure of 30-40 MPa and a temperature of 45-55℃. (3) Supercritical carbon dioxide fluid carrying frog seed oil is introduced into a separation vessel, and frog seed oil is recovered after depressurization.
9. A composition for preventing and improving depression, characterized in that, Its active ingredients include frog seeds and their extracts, as well as one or more pharmaceutically or food-grade carriers, diluents or excipients.
10. The composition according to claim 9, characterized in that, The composition contains 5-45% by weight of water extract of frog seeds or 10-60% by volume of oil extracted from frog seeds.