Composition for treating or improving depression and anxiety as well as preparation method and application thereof

Abstract

The invention provides a composition for treating or improving depression and anxiety. The composition comprises the following raw materials in parts by weight: 200-1000 parts of fish oil, 8-60 parts of vitamins and 10-100 parts of minerals. Wherein the content of ethyl eicosapentaenoate (EPA-EE) in the fish oil is greater than or equal to 80% w / w; the vitamin is selected from at least one of vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D and folic acid; the mineral substance is selected from at least one of calcium, magnesium, zinc, iron, potassium and phosphorus. The invention also provides a preparation method and application of the composition. According to the fish oil composition provided by the invention, through scientific active component compatibility and a specific preparation process, while stable coexistence of core components such as fish oil and the like is ensured, a clinical curative effect equivalent to that of a conventional antidepressant drug is realized, and a new high-quality choice is provided for management of mood disorders such as depression and anxiety.

Description

TECHNICAL FIELD

[0001] The present application relates to a composition for treating or ameliorating depression, anxiety and a preparation method and use thereof. BACKGROUND

[0002] Depression, as a common mental disorder, has diverse and complex symptoms involving mood, cognition, physiology and other aspects of functional disorders. The main manifestations are persistent low mood, loss of interest, often accompanied by slow thinking, sleep problems, decreased executive function and impaired social skills. In severe cases, patients may have suicidal thoughts and behaviors. Anxiety, also known as anxiety neurosis, is mainly characterized by intense, excessive and persistent worry and fear, accompanied by symptoms such as rapid heart rate and sleep difficulties. Depression and anxiety have become common emotional disorders in the international community in recent years. According to statistics, the annual prevalence rate of anxiety disorders among residents aged 18 and above in China is about 5.0%, and the lifetime prevalence rate is about 7.6%. There are more than 95 million patients with depression, and the lifetime prevalence rate of depression is about 6.9%, with an annual incidence rate of about 3.6%. At present, due to the increasing incidence of depression and anxiety worldwide, it has had an impact on society and human health.

[0003] Currently, the treatment of emotional disorders such as depression and anxiety is mainly focused on the use of drugs. Antidepressants include classic antidepressants and new antidepressants. Selective serotonin reuptake inhibitors (SSRIs) and serotonin and norepinephrine reuptake inhibitors (SNRIs) are the most common first-line antidepressants in clinical practice. In addition, monoamine oxidase inhibitors (MAOIs), tricyclic antidepressants (TCAs) and atypical antidepressants also have antidepressant effects. Although existing antidepressants play an important role in treatment, they have some limitations, such as slow onset, high risk of relapse after discontinuation, and side effects, which to some extent weaken their clinical efficacy. Therefore, it is particularly urgent and necessary to explore an effective and relatively safe treatment strategy to improve the symptoms and quality of life of patients with emotional disorders such as depression and anxiety.

[0004] Fish oil is an oil extracted from fish, rich in Omega-3 unsaturated fatty acids, mainly including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and has been widely used in the fields of medicine and health. At present, high-purity and high-concentration fish oil preparations have been approved for the treatment of specific diseases such as hypertriglyceridemia. In addition, fish oil has been widely used in health products or functional foods due to its basic health care functions in maintaining cardiovascular and cerebrovascular health and assisting in improving memory.

[0005] Although fish oil has been successfully applied in the aforementioned fields, there are currently no reports on its use in treating or improving depression and anxiety, particularly its scientific formulation with other active ingredients to develop products with clearly defined antidepressant and anti-anxiety functions. Therefore, developing a fish oil composition with proven efficacy against depression and anxiety is of great significance for enriching clinical and consumer choices. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention aims to provide a highly stable fish oil composition that can be used to treat or improve mood disorders such as depression and anxiety, with few adverse reactions. Simultaneously, this composition also helps improve sleep, maintain healthy blood lipid levels, and provides antioxidant benefits.

[0007] This invention provides a composition for treating or improving depression and anxiety, comprising the following raw materials in the indicated weight ratios: Fish oil 200-1000 parts, vitamins 8-60 parts, minerals 10-100 parts; Among them, the content of eicosapentaenoic acid ethyl ester (EPA-EE) in fish oil is ≥80% w / w; The vitamin is selected from at least one of vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, and folic acid; The mineral is selected from at least one of calcium, magnesium, zinc, iron, potassium, and phosphorus.

[0008] The folic acid in question is L-5-methyltetrahydrofolate calcium.

[0009] The calcium in the mineral is selected from at least one of calcium carbonate, calcium chloride, calcium acetate, calcium sulfate, calcium citrate, calcium gluconate, calcium lactate, and calcium hydrogen phosphate; the magnesium is selected from at least one of magnesium oxide, magnesium sulfate, magnesium carbonate, magnesium glycinate, magnesium gluconate, magnesium citrate, and magnesium L-threonate; and the zinc is selected from at least one of zinc oxide, zinc sulfate, zinc glycinate, zinc citrate, zinc gluconate, zinc lactate, and zinc acetate.

[0010] The raw materials also contain betaine, and the weight ratio of the raw materials is as follows: Fish oil 200-1000 parts, vitamins 8-60 parts, betaine 5-30 parts, minerals 10-100 parts; Fish oil contains ≥85% w / w of eicosapentaenoic acid ethyl ester (EPA-EE); The betaine is selected from anhydrous betaine.

[0011] Further, the composition comprises raw materials in the following weight ratios: Fish oil 200-1000 parts, L-5-methyltetrahydrofolate calcium 1-10 parts, vitamin B2 5-15 parts, vitamin B6 1-10 parts, vitamin B12 0-5 parts, vitamin C 1-10 parts, vitamin D3 0.005-0.1 parts, betaine 5-30 parts, calcium (as elemental calcium) 5-25 parts, magnesium (as elemental magnesium) 15-50 parts, zinc (as elemental zinc) 2-15 parts; Among them, the content of eicosapentaenoic acid ethyl ester (EPA-EE) in fish oil is ≥90%w / w.

[0012] Further, the composition comprises raw materials in the following weight ratios: Fish oil 300-600 parts, L-5-methyltetrahydrofolate calcium 2-7 parts, vitamin B2 6-10 parts, vitamin B6 3.75-6 parts, vitamin B12 0.25-0.4 parts, vitamin C 2-3.2 parts, vitamin D3 0.01-0.05 parts, betaine 10-20 parts, calcium as elemental calcium 9-14 parts, magnesium as elemental magnesium 25-34 parts, zinc glycine as elemental zinc 6-9 parts; Among them, the content of eicosapentaenoic acid ethyl ester (EPA-EE) in fish oil is ≥90%w / w.

[0013] Further, the composition comprises raw materials in the following weight ratios: Fish oil 334-492 parts, L-5-methyltetrahydrofolate calcium 2.05-6.38 parts, vitamin B2 6.25-8.7 parts, vitamin B6 3.75-5.2 parts, vitamin B12 0.25-0.35 parts, vitamin C 2.0-2.75 parts, vitamin D3 0.01-0.03 parts, betaine 10-12 parts, calcium (as elemental calcium) 9.79-11.73 parts, magnesium (as elemental magnesium) 25-30.15 parts, zinc (as elemental zinc) 6.25-6.74 parts; The fish oil contains ethyl eicosapentaenoic acid (EPA-EE) at a content ≥90% w / w. Further, the composition comprises the following raw materials in the indicated weight ratios: Fish oil 380 parts, L-5-methyltetrahydrofolate calcium 3.5 parts, vitamin B2 8.7 parts, vitamin B6 5.2 parts, vitamin B12 0.35 parts, vitamin C 2.43 parts, vitamin D3 0.02625 parts, betaine 10 parts, calcium carbonate (calculated as elemental calcium) 10 parts, magnesium oxide (calculated as elemental magnesium) 30.15 parts, zinc glycine (calculated as elemental zinc) 6.74 parts; or Fish oil 380 parts, L-5-methyltetrahydrofolate calcium 2.05 parts, vitamin B2 6.25 parts, vitamin B6 3.75 parts, vitamin B12 0.25 parts, vitamin C 2.0 parts, vitamin D3 0.01875 parts, betaine 10 parts, calcium carbonate (calculated as elemental calcium) 9.79 parts, magnesium oxide (calculated as elemental magnesium) 25 parts, zinc glycine (calculated as elemental zinc) 6.25 parts; or Fish oil 367 parts, L-5-methyltetrahydrofolate calcium 3.5 parts, vitamin B2 8.7 parts, vitamin B6 5.2 parts, vitamin B12 0.35 parts, vitamin C 2.75 parts, vitamin D3 0.02625 parts, betaine 12 parts, calcium carbonate (as elemental calcium) 11.73 parts, magnesium oxide (as elemental magnesium) 30.15 parts, zinc glycine (as elemental zinc) 6.74 parts; or Fish oil 334 parts, L-5-methyltetrahydrofolate calcium 2.05 parts, vitamin B2 6.25 parts, vitamin B6 3.75 parts, vitamin B12 0.25 parts, vitamin C 2.0 parts, vitamin D3 0.01875 parts, betaine 10 parts, calcium carbonate (calculated as elemental calcium) 9.79 parts, magnesium oxide (calculated as elemental magnesium) 25 parts, zinc glycine (calculated as elemental zinc) 6.25 parts; or Fish oil 492 parts, L-5-methyltetrahydrofolate calcium 6.38 parts, vitamin B2 6.25 parts, vitamin B6 3.75 parts, vitamin B12 0.25 parts, vitamin C 2.0 parts, vitamin D3 0.01875 parts, betaine 10 parts, calcium carbonate (calculated as elemental calcium) 9.79 parts, glycine magnesium (calculated as elemental magnesium) 25 parts, glycine zinc (calculated as elemental zinc) 6.25 parts; or The composition includes 445 parts fish oil, 3.75 parts L-5-methyltetrahydrofolate calcium, 6.25 parts vitamin B2, 3.75 parts vitamin B6, 0.25 parts vitamin B12, 2.0 parts vitamin C, 0.01875 parts vitamin D3, 10 parts betaine, 9.79 parts calcium carbonate (calculated as elemental calcium), 25 parts magnesium glycine (calculated as elemental magnesium), and 6.25 parts zinc glycine (calculated as elemental zinc); wherein the fish oil contains ≥90% w / w eicosapentaenoic acid ethyl ester (EPA-EE).

[0014] The composition of the present invention is a formulation prepared by adding acceptable excipients or auxiliary ingredients to the raw materials described herein as functional components.

[0015] The auxiliary components include wax and / or oil; the wax is selected from beeswax; the oil is selected from at least one of lemon essential oil, sunflower seed oil, soybean oil, and olive oil. The preparation is a capsule, emulsion, tablet or granule.

[0016] The present invention provides a method for preparing the aforementioned composition, comprising the following steps: a. Dissolving the colloid: Add purified water, gelatin, glycerin, and pigment to the dissolving tank, heat to 50-80℃ and stir until the colloid is completely dissolved and the mixture is homogeneous; b. Ingredients: Take at least 1 / 3 of the formula amount of fish oil, add beeswax, heat to 60-80℃, the beeswax will dissolve, add the remaining fish oil, cool to room temperature, and obtain thickened fish oil; take the formula amount of L-5-methyltetrahydrofolate calcium, vitamin B12, vitamin B2, vitamin B6, vitamin C, betaine, and minerals in the same container, mix them, and obtain mixed solid material; pulverize the mixed solid material with a high-speed pulverizer for at least 20 seconds, and sieve the pulverized material with a sieve size of 40 mesh. The material that cannot be sieved is ground with a mortar and pestle; add the sieved material and the ground material to the thickened fish oil, then add vitamin D3 and auxiliary component oil, and homogenize using a high-shear dispersing emulsifier for at least 1 minute to obtain a suspension, which is the contents; c. Shot pressing, shaping, drying, and shot selection; Preferably, the heating temperature in step b is 65-70℃; the pulverizing time of the high-speed pulverizer is not less than 30s.

[0017] The present invention provides the use of the composition in the preparation of a medicament for treating and / or improving depression and anxiety; preferably, the depression is mild to moderate depression.

[0018] The present invention provides the use of the described composition in the preparation of dietary supplements or health foods that help improve sleep.

[0019] This invention provides the use of the described composition in the preparation of dietary supplements or health foods that help maintain healthy blood lipid levels.

[0020] This invention provides the use of the described composition in the preparation of dietary supplements or health foods that contribute to antioxidant activity.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. Clear and significant efficacy: Animal model experiments and human clinical trials have confirmed that the composition provided by this invention has significant effects in improving depressive and anxious behaviors, comparable to conventional antidepressant drugs. At the same time, due to its natural ingredients, it avoids the potential side effects and dependence of chemical drugs, providing a safer solution for treating or improving mood disorders such as depression and anxiety.

[0022] 2. Good product safety: Animal toxicity tests show that the composition provided by this invention is significantly safe and suitable for long-term use.

[0023] 3. The product has excellent stability: Through scientific ingredient compatibility and specific preparation process, this invention effectively protects the stable coexistence of fish oil and other active ingredients, ensuring that the content of the product remains stable and uniform during long-term storage, thereby guaranteeing the quality and efficacy of the final product.

[0024] 4. Excellent user experience and high compliance: This invention uses soft capsule encapsulation technology to encapsulate active ingredients such as fish oil, and with the addition of flavor adjustment, it successfully masks unpleasant odors, improves the taste and palatability of the product, and makes it easier for patients to accept. Detailed Implementation

[0025] The raw materials and equipment used in the specific embodiments of the present invention are all known products, obtained by purchasing commercially available products.

[0026] The term “room temperature” as used in this article refers to a temperature ranging from 5°C to 35°C, such as 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, or 35°C.

[0027] The term “EPA-EE” used in this article refers to ethyl eicosate pentanoate.

[0028] Example 1: Preparation of fish oil soft capsules formula:

[0029]

[0030] Rubber composition: gelatin, glycerin, purified water, pigments (red iron oxide premix, black iron oxide premix).

[0031] Preparation method: a. Glue dissolving: Weigh out the prescribed amounts of purified water, gelatin, glycerin, and pigment, and add them to a water bath dissolving tank. Heat and stir until the temperature reaches 80°C. After the colloid is completely dissolved, stir evenly and evacuate to -0.08 MPa using a vacuum pump. When there are no more bubbles in the colloid solution, maintain the temperature at 60°C ± 5°C for later use. b. Ingredients: Take 1 / 3 of the formula amount of fish oil, add the formula amount of beeswax, heat at 70℃ until the beeswax melts, add the remaining fish oil, cool to room temperature, and obtain thickened fish oil; take the formula amounts of L-5-methyltetrahydrofolate calcium, vitamin B12, vitamin B2, vitamin B6, vitamin C, betaine, and minerals in the same container, mix them, and obtain mixed solid material; pulverize the mixed solid material with a high-speed pulverizer for 30 seconds, and pass the pulverized material through a 40-mesh sieve. The material that cannot be sieved is ground with a mortar and pestle; add the sieved material and the ground material to the thickened fish oil, then add vitamin D3 and auxiliary component oil, and homogenize using a high-shear dispersing emulsifier for 2 minutes to obtain a suspension, which is the contents.

[0032] c. Squash pressing, shaping, drying, and pellet selection: Add the obtained contents into the soft capsule machine, adjust the contents volume, press the soft capsules to obtain fish oil soft capsules, place the obtained soft capsules in a single-layer shaping drum for shaping, drying, and pellet selection to obtain the final product.

[0033] Example 2: Preparation of fish oil soft capsules formula:

[0034]

[0035] Rubber composition: gelatin, glycerin, purified water, pigments (red iron oxide premix, black iron oxide premix).

[0036] The preparation method is the same as in Example 1.

[0037] Example 3: Preparation of fish oil soft capsules formula:

[0038]

[0039] Rubber composition: gelatin, glycerin, purified water, pigments (red iron oxide premix, black iron oxide premix).

[0040] The preparation method is the same as in Example 1.

[0041] Example 4: Preparation of fish oil soft capsules formula:

[0042]

[0043] Rubber composition: gelatin, glycerin, purified water, pigments (red iron oxide premix, black iron oxide premix).

[0044] The preparation method is the same as in Example 1.

[0045] Example 5: Preparation of fish oil soft capsules formula:

[0046]

[0047] Rubber composition: gelatin, glycerin, purified water, pigments (caramel color, carmine red).

[0048] The preparation method is the same as in Example 1.

[0049] Example 6: Preparation of fish oil soft capsules formula:

[0050]

[0051] Rubber composition: gelatin, glycerin, purified water, pigments (caramel color, carmine red).

[0052] The preparation method is the same as in Example 1.

[0053] Example 7 Sensory evaluation of fish oil soft capsules The fish oil soft capsules prepared according to the present invention were subjected to sensory evaluation, and the results are shown in Table 1.

[0054] Table 1 Sensory evaluation of fish oil soft capsules Sample Sensory evaluation Example 1 No fishy smell Example 2 No fishy smell Example 3 No fishy smell Example 4 No fishy smell Example 5 No fishy smell Example 6 No fishy smell The fish oil soft capsules prepared by this invention effectively reduce the fishy smell through the reasonable combination of various components. The soft capsules further reduce the release of the fishy smell, thereby effectively masking the fishy smell and making it more acceptable to the user.

[0055] Example 8: Stability Study of Fish Oil Soft Capsules 1. Influencing Factors Experiment Fish oil soft capsules were placed under high temperature (60℃±2℃), high humidity (90%RH±5%RH / 25℃), and light (4500lx±500lx) conditions. Samples were taken at 0 days, 10 days, and 30 days to detect the content of EPA-EE and L-5-methyltetrahydrofolate calcium.

[0056] EPA-EE content determination method: The sample was determined by gas chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0521). Chromatographic conditions: quartz capillary column (Agilent DB-WAX, 0.32 mm × 30 m, 0.5 μm) with polyethylene glycol as stationary phase; initial column temperature 170 °C, held for 2 min, then increased to 240 °C at a rate of 2 °C / min, held for 8 min; injection port temperature 250 °C, detector (FID) temperature 270 °C, carrier gas nitrogen; split ratio 20:1, injection volume 1 μl. Calculation was performed using peak area according to the internal standard method.

[0057] Method for determining the content of L-5-methyltetrahydrofolate calcium: The determination was performed according to high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512). Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase (Agilent ZORBAX SB-C18 4.6×250mm, 5µm column, or a column with equivalent performance); phosphate buffer was used as mobile phase A; phosphate buffer-methanol (65:35) (7.8g of sodium dihydrogen phosphate dihydrate was weighed and added to 1000ml of ultrapure water, mixed well, 650ml was taken, 350ml of methanol was added, mixed well, and the pH was adjusted to 8.0 with 32% sodium hydroxide solution) was used as mobile phase B, and gradient elution was performed according to the table below; the detection wavelength was 280nm; the flow rate was 1.1ml per minute; the column temperature was 32℃; and the injection volume was 10μl.

[0058]

[0059] The stability results are shown in Table 2.

[0060] Table 2. Stability data of fish oil soft capsules under different conditions

[0061]

[0062] 2. Long-term stability test Fish oil soft capsules were placed at 25℃±2℃ and 60%RH±5%RH, and samples were taken at 0 days, 3 months, and 6 months. The contents of EPA-EE and L-5-methyltetrahydrofolate calcium were determined according to the detection conditions in the influencing factor test. The results are shown in Table 3.

[0063] Table 3. Long-term stability test data of fish oil soft capsules

[0064]

[0065] Fish oil is rich in Omega-3 unsaturated fatty acids, which are prone to oxidative degradation at room temperature. L-5-methyltetrahydrofolate calcium, the active form of folic acid, has high bioavailability but is easily oxidized and has poor stability. Table 2 shows that after 30 days of storage under high temperature, high humidity, and light conditions, the EPA-EE content and L-5-methyltetrahydrofolate calcium content of the fish oil soft capsules prepared according to this invention remained essentially unchanged, demonstrating the good stability of the fish oil soft capsules prepared according to this invention. Furthermore, long-term stability tests (Table 3) also confirm that the fish oil soft capsules prepared according to this invention have excellent stability and are suitable for long-term storage.

[0066] Example 9: Content Uniformity Test The content uniformity test was conducted according to General Chapter 0941, "Method for Testing Content Uniformity," of the 2025 edition of the Chinese Pharmacopoeia. Ten fish oil soft capsules were randomly selected, and the content of L-5-methyltetrahydrofolate calcium was determined. The relative standard deviation (RSD) of the 10 single-dose content determination results was calculated. If the RSD was not greater than 5.0%, the content uniformity of the sample was considered to meet the requirements. The results are shown in Table 4.

[0067] Table 4 Results of content uniformity test Sample RSD Conclusion Example 1 3.0% Content uniformity meets the requirements Example 2 2.9% Content uniformity meets the requirements Example 3 3.2% Content uniformity meets the requirements Example 4 3.0% Content uniformity meets the requirements Example 5 2.1% Content uniformity meets the requirements Example 6 2.5% Content uniformity meets the requirements Example 10 Animal toxicity test 1. Experimental objective: To observe the toxic effects of the fish oil capsules of this invention administered orally to SD rats for 28 consecutive days, and to determine the dose level at which no clinical adverse reactions were observed, so as to provide a reference for the safety of long-term clinical use.

[0068] 2. Experimental Materials and Methods Test sample: The contents of fish oil soft capsules prepared in Example 4 of this invention.

[0069] Animals and grouping: Eighty SD rats, half male and half female, were randomly divided into four groups according to their body weight: solvent control group (sunflower seed oil), low-dose group (0.47 g / kg), medium-dose group (1.4 g / kg), and high-dose group (4.2 g / kg) of the test sample.

[0070] Dosing regimen: Rats in each group were administered the test sample or solvent at various concentrations orally via gavage once daily. For four consecutive weeks, the gavage volume was 4 mL / kg.

[0071] Monitoring Indicators: All animals underwent daily clinical observation during the experiment. Body Weight: Weighed once before the experiment and once weekly during the experiment. Food Intake: Food intake was measured weekly after the start of the experiment. Ophthalmological Examination: All animals were examined once after grouping, and the solvent control group and high-dose group were examined once each on day 27 of the experiment. Urine: Measured once on day 27 of the experiment. On day 29 of the experiment, all animals were dissected, and blood was collected for hematological, coagulation, and blood biochemical tests. Gross necropsy, organ weighing, and histopathological examination were performed. Bone marrow cell differential counts were performed on the solvent control group and high-dose group animals at the end of the experiment.

[0072] 3. Test Results Under the conditions of this experiment, SD rats were administered fish oil soft capsule contents orally for 4 consecutive weeks at doses of 0.47 g / kg, 1.4 g / kg, and 4.2 g / kg. No animals were near death or died during the experiment. No significant toxic reactions were observed in the animals based on clinical observation, body weight, food intake, ophthalmological examination, hematological indicators, blood coagulation indicators, blood biochemical indicators, urine indicators, bone marrow cell examination, gross necropsy, organ weight, and histopathological examination at each dose of fish oil soft capsule contents.

[0073] In summary, under the conditions of this experiment, SD rats were administered the contents of the fish oil soft capsules of this invention via oral gavage for 4 consecutive weeks, and the no-adverse-effect level (NOAEL) was 4.2 g / kg (equivalent to 116 times the recommended human dose). These results demonstrate that the fish oil product of this invention has good safety within the tested dosage range.

[0074] Example 11 Evaluation of the antidepressant effect of fish oil soft capsules using a zebrafish model. Test sample: The contents of the fish oil soft capsules prepared in Example 4 of this invention.

[0075] Experimental animals and grouping: Wild-type AB strain zebrafish (5 days post-fertilization) were randomly divided into six groups: normal control group, model control group, positive drug control group (sertraline, 100 ng / mL), low-dose test sample group (31.2 μg / mL), medium-dose test sample group (62.5 μg / mL), and high-dose test sample group (125 μg / mL).

[0076] Model establishment: Zebrafish were placed in 6-well plates, with 30 fish per well and a volume of 3 mL per well. Except for the normal control group, all other groups were given reserpine (5 μg / mL, prepared as stock solution with DMSO) to establish a depression model.

[0077] Test sample preparation: The contents of the fish oil soft capsules were prepared into a 200 mg / mL stock solution in 100% DMSO, sonicated for 5 minutes, and then administered as a water solution at the corresponding final concentrations (31.2 μg / mL, 62.5 μg / mL, and 125 μg / mL). Sertraline, the positive control drug, was prepared into a 10 mg / mL stock solution in ultrapure water, sonicated for 5 minutes, and then administered as a water solution at a final concentration of 100 ng / mL. After treatment at 28℃ for 1 day, 10 zebrafish were randomly selected from each experimental group and transferred to a 24-well plate (1 zebrafish / well, 1000 μL / well). The proportion of time the zebrafish spent moving in the open field was measured using a behavioral analysis system to evaluate the antidepressant activity of the fish oil soft capsules.

[0078] Statistical analysis: Results are expressed as Mean ± SE. Statistical analysis was performed using SPSS software. A p-value < 0.05 was considered statistically significant compared to the model control group.

[0079] Table 5. Evaluation results of the antidepressant effect of fish oil soft capsules in a zebrafish model.

[0080] As shown in Table 5, in the zebrafish animal model, the fish oil composition of the present invention dose-dependently increased the proportion of open field exercise time, and the medium and high dose groups showed significant differences compared with the model group, indicating that the fish oil of the present invention has antidepressant effects. Among them, the effect of the high dose group was close to that of the positive control drug sertraline.

[0081] Example 12: Human clinical study on the efficacy and safety of fish oil soft capsules in improving mild to moderate depression. Objective: To evaluate the efficacy and safety of the fish oil soft capsules of this invention compared with fluoxetine hydrochloride capsules in patients with mild to moderate depression.

[0082] Participants: Aged 18-65 years (inclusive), gender not limited; meeting the diagnostic criteria for depressive episode in the Diagnostic and Statistical Manual of Mental Disorders (5th Edition) (DSM-5); total HAMD score ≥8 and <24; HAMD 17 item score ≥2; CGI-S score ≥3; understanding and voluntarily participating in this trial, and signing an informed consent form.

[0083] Study Protocol: This study is a randomized, double-blind, double-dummy, positive-drug parallel-controlled, multicenter clinical trial. Experimental Group: Subjects took 6 fish oil soft capsules (prepared according to Example 4 of this invention, divided into two doses) daily, along with 1 fluoxetine hydrochloride capsule analog, for 8 consecutive weeks. During the trial, subjects were not permitted to use any psychotropic drugs other than the study drug or receive any behavioral interventions. Positive-Drug Control Group: Subjects took 1 fluoxetine hydrochloride capsule (20 mg) daily, along with 6 fish oil soft capsule analogs (divided into two doses) daily, for 8 consecutive weeks. During the trial, subjects were not permitted to use any psychotropic drugs other than the study drug or receive any behavioral interventions.

[0084] Primary efficacy endpoint: Change in total Hamilton Depression Rating Scale (HAMD 17 items) score from baseline after 8 weeks of double-blind treatment.

[0085] Safety evaluation indicators: (1) adverse events; (2) laboratory tests (complete blood count, complete urine count, blood biochemistry, thyroid function, blood pregnancy test (for women of childbearing age)); (3) 12-lead electrocardiogram, heart rate variability analysis (HRV instrument); (4) vital signs (including body temperature, blood pressure, pulse); (5) physical examination; Columbia Suicide Severity Rating Scale (C-SSRS) assessment.

[0086] Statistical analysis methods: SAS statistical analysis software was used. All statistical tests were two-tailed tests, and a p-value less than or equal to 0.05 was considered statistically significant.

[0087] The test results are shown in Table 6.

[0088] Table 6 Clinical Trial Efficacy Data

[0089]

[0090] Table 6 shows the results. The total HAMD17 score in both the experimental group (fish oil soft capsules of this invention) and the positive control group (fluoxetine hydrochloride capsules) decreased significantly from baseline, with statistically significant differences (P<0.05). After 8 weeks of treatment, the ΔHAMD17 score decreased by 9.29±4.31 points from baseline in the fish oil group and by 8.72±4.30 points in the positive control group. These results indicate that the fish oil composition of this invention has a clear efficacy in improving mild to moderate depression, and its effect is comparable to that of the classic antidepressant fluoxetine hydrochloride.

[0091] Of particular note is that the fish oil composition of the present invention, while achieving similar therapeutic effects to chemically synthesized drugs, also has the potential advantages of being naturally sourced and having high safety, which helps to improve patients' willingness to undergo treatment and their long-term adherence, and has a positive significance for improving the quality of life of patients with depression.

[0092] In summary, this invention provides a fish oil composition that combines proven antidepressant efficacy with high safety. Through scientific formulation of active ingredients and a specific preparation process, the core components, such as fish oil, are stably coexisted while achieving clinical efficacy comparable to conventional antidepressants. Furthermore, by utilizing dosage form design and flavor control, the palatability and patient compliance of the product are significantly improved. The fish oil composition provided by this invention offers a new and high-quality option for the management of mood disorders such as depression and anxiety.

Claims

1. A composition for treating or ameliorating depression, anxiety, characterized in that, The raw materials include the following weight proportions: fish oil 200-1000 parts, vitamins 8-60 parts, and minerals 10-100 parts; The fish oil contains ≥80% w / w of eicosapentaenoic acid ethyl ester (EPA-EE); The vitamins are at least one selected from the group consisting of vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, and folic acid; The minerals are at least one selected from the group consisting of calcium, magnesium, zinc, iron, potassium, and phosphorus.

2. The composition of claim 1, wherein, The folic acid is L-5-methyltetrahydrofolic acid calcium; The calcium in the minerals is at least one selected from the group consisting of calcium carbonate, calcium chloride, calcium acetate, calcium sulfate, calcium citrate, calcium gluconate, calcium lactate, and calcium hydrogen phosphate; The magnesium in the minerals is at least one selected from the group consisting of magnesium oxide, magnesium sulfate, magnesium carbonate, magnesium glycinate, magnesium gluconate, magnesium citrate, and L-magnesium threonate; The zinc in the minerals is at least one selected from the group consisting of zinc oxide, zinc sulfate, zinc glycinate, zinc citrate, zinc gluconate, zinc lactate, and zinc acetate.

3. The composition according to claim 1 or 2, characterized in that, The raw materials further include betaine, and the weight proportions of the raw materials are as follows: fish oil 200-1000 parts, vitamins 8-60 parts, betaine 5-30 parts, and minerals 10-100 parts; The fish oil contains ≥85% w / w of eicosapentaenoic acid ethyl ester (EPA-EE); and the betaine is anhydrous betaine.

4. The composition of claim 3, wherein, The raw materials include the following weight proportions: fish oil 200-1000 parts, L-5-methyltetrahydrofolic acid calcium 1-10 parts, vitamin B2 5-15 parts, vitamin B6 1-10 parts, vitamin B12 0-5 parts, vitamin C 1-10 parts, vitamin D3 0.005-0.1 part, betaine 5-30 parts, calcium 5-25 parts in terms of elemental calcium, magnesium 15-50 parts in terms of elemental magnesium, and zinc 2-15 parts in terms of elemental zinc; The fish oil contains ≥90% w / w of eicosapentaenoic acid ethyl ester (EPA-EE).

5. The composition of claim 3, wherein The raw materials include the following weight proportions: fish oil 300-600 parts, L-5-methyltetrahydrofolic acid calcium 2-7 parts, vitamin B2 6-10 parts, vitamin B6 3.75-6 parts, vitamin B12 0.25-0.4 part, vitamin C 2-3.2 parts, vitamin D3 0.01-0.05 part, betaine 10-20 parts, calcium 9-14 parts in terms of elemental calcium, magnesium 25-34 parts in terms of elemental magnesium, and zinc 6-9 parts in terms of elemental zinc; The fish oil contains ≥90% w / w of eicosapentaenoic acid ethyl ester (EPA-EE).

6. The composition of claim 3, wherein The raw materials include the following weight proportions: fish oil 334-492 parts, L-5-methyltetrahydrofolic acid calcium 2.05-6.38 parts, vitamin B2 6.25-8.7 parts, vitamin B6 3.75-5.2 parts, vitamin B12 0.25-0.35 part, vitamin C 2.0-2.75 parts, vitamin D3 0.01-0.03 part, betaine 10-12 parts, calcium 9.79-11.73 parts in terms of elemental calcium, magnesium 25-30.15 parts in terms of elemental magnesium, and zinc 6.25-6.74 parts in terms of elemental zinc; The content of eicosapentaenoic acid ethyl ester (EPA-EE) in the fish oil is greater than or equal to 90% w / w.

7. The composition of claim 3, wherein The raw materials include the following weight proportions: 380 parts of fish oil, 3.5 parts of L-5-methyltetrahydrofolate calcium, 8.7 parts of vitamin B2, 5.2 parts of vitamin B6, 0.35 parts of vitamin B12, 2.43 parts of vitamin C, 0.02625 parts of vitamin D3, 10 parts of betaine, 10 parts of calcium carbonate in terms of elemental calcium, 30.15 parts of magnesium oxide in terms of elemental magnesium, and 6.74 parts of zinc glycinate in terms of elemental zinc; or 380 parts of fish oil, 2.05 parts of L-5-methyltetrahydrofolate calcium, 6.25 parts of vitamin B2, 3.75 parts of vitamin B6, 0.25 parts of vitamin B12, 2.0 parts of vitamin C, 0.01875 parts of vitamin D3, 10 parts of betaine, 9.79 parts of calcium carbonate in terms of elemental calcium, 25 parts of magnesium oxide in terms of elemental magnesium, and 6.25 parts of zinc glycinate in terms of elemental zinc; or 367 parts of fish oil, 3.5 parts of L-5-methyltetrahydrofolate calcium, 8.7 parts of vitamin B2, 5.2 parts of vitamin B6, 0.35 parts of vitamin B12, 2.75 parts of vitamin C, 0.02625 parts of vitamin D3, 12 parts of betaine, 11.73 parts of calcium carbonate in terms of elemental calcium, 30.15 parts of magnesium oxide in terms of elemental magnesium, and 6.74 parts of zinc glycinate in terms of elemental zinc; or 367 parts of fish oil, 3.5 parts of L-5-methyltetrahydrofolate calcium, 8.7 parts of vitamin B2, 5.2 parts of vitamin B6, 0.35 parts of vitamin B12, 2.75 parts of vitamin C, 0.02625 parts of vitamin D3, 12 parts of betaine, 11.73 parts of calcium carbonate in terms of elemental calcium, 30.15 parts of magnesium oxide in terms of elemental magnesium, and 6.74 parts of zinc glycinate in terms of elemental zinc; or 334 parts of fish oil, 2.05 parts of L-5-methyltetrahydrofolate calcium, 6.25 parts of vitamin B2, 3.75 parts of vitamin B6, 0.25 parts of vitamin B12, 2.0 parts of vitamin C, 0.01875 parts of vitamin D3, 10 parts of betaine, 9.79 parts of calcium carbonate in terms of elemental calcium, 25 parts of magnesium oxide in terms of elemental magnesium, and 6.25 parts of zinc glycinate in terms of elemental zinc; or 492 parts of fish oil, 6.38 parts of L-5-methyltetrahydrofolate calcium, 6.25 parts of vitamin B2, 3.75 parts of vitamin B6, 0.25 parts of vitamin B12, 2.0 parts of vitamin C, 0.01875 parts of vitamin D3, 10 parts of betaine, 9.79 parts of calcium carbonate in terms of elemental calcium, 25 parts of magnesium glycinate in terms of elemental magnesium, and 6.25 parts of zinc glycinate in terms of elemental zinc; or 445 parts of fish oil, 3.75 parts of L-5-methyltetrahydrofolate calcium, 6.25 parts of vitamin B2, 3.75 parts of vitamin B6, 0.25 parts of vitamin B12, 2.0 parts of vitamin C, 0.01875 parts of vitamin D3, 10 parts of betaine, 9.79 parts of calcium carbonate in terms of elemental calcium, 25 parts of magnesium glycinate in terms of elemental magnesium, and 6.25 parts of zinc glycinate in terms of elemental zinc; wherein the content of eicosapentaenoic acid ethyl ester (EPA-EE) in the fish oil is greater than or equal to 90% w / w.

8. The composition according to any one of claims 1 to 7, characterized in that, It is prepared from the raw material as a functional ingredient, and acceptable auxiliary ingredients or auxiliary ingredients; preferably, the auxiliary ingredients include wax and / or oil; the wax is selected from beeswax; the oil is selected from at least one of lemon essential oil, sunflower oil, soybean oil, olive oil; The preparation is a capsule, emulsion, tablet or granule.

9. Process for the preparation of a composition according to claim 8, characterized in that, It comprises the following steps: a. Gelation: add purified water, gelatin, glycerol and pigment into a gelation tank, heat and stir to 50-80℃, dissolve all the colloids and stir uniformly; b. Blending: take no less than 1 / 3 of the formula amount of fish oil, add beeswax, heat to 60-80℃, dissolve the beeswax, add the remaining fish oil, cool to room temperature, and obtain thickened fish oil; take the formula amount of L-5-methyltetrahydrofolate calcium, vitamin B12, vitamin B2, vitamin B6, vitamin C, betaine and minerals in the same container, mix to obtain a mixture of solid materials; grind the mixture of solid materials with a high-speed grinder for no less than 20s, sieve the ground materials with a 40-mesh sieve, and grind the materials that cannot pass through the sieve with a mortar; add the sieved materials and ground materials into the thickened fish oil, add vitamin D3 and auxiliary ingredient oil, and use a high-shear dispersion emulsifier to homogenize for no less than 1min to obtain a suspension, which is the content; c. Pill pressing, shaping, drying and pill selecting; Preferably, the heating temperature in step b is 65-70℃, and the high-speed grinder grinding time is no less than 30s.

10. Use of the composition of any one of claims 1-8 in the preparation of a medicament for treating and / or improving depression, anxiety; preferably, the depression is mild to moderate depression.

11. Use of the composition of any one of claims 1-8 in the preparation of a dietary supplement or health food for improving sleep, maintaining blood lipid health level or antioxidant.

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