Application of p. tianschanicus extract in preparation of antidepressant drugs
The preparation of antidepressant drugs using Viola tianshanense extract has solved the problems of significant side effects and slow efficacy of traditional drugs, achieving significant antidepressant effects and low-cost production.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- SHIHEZI UNIVERSITY
- Filing Date
- 2024-08-22
- Publication Date
- 2026-07-24
AI Technical Summary
Existing antidepressants have significant side effects and slow onset of therapeutic effects.
Viola tianshanensis extract, particularly Viola tianshanensis extract obtained by ethanol extraction from the dried whole herb, is used to prepare antidepressant drugs, including tablets, capsules, pellets, sustained-release tablets, granules, or oral liquid preparations, supplemented with pharmaceutically acceptable excipients.
Viola tianshanense extract exhibits significant antidepressant activity, can significantly improve depressive symptoms, has few side effects, is suitable for large-scale production, and is low in cost.
Smart Images
Figure CN118831124B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to the application of Viola tianshanense extract in the preparation of antidepressant drugs. Background Technology
[0002] Depression, as a widespread mental health disorder, has attracted widespread attention due to its high incidence rate. The core characteristics of depression include persistent low mood, loss of interest, reduced pleasure, and possibly accompanying sleep disturbances, changes in appetite, difficulty concentrating, and decreased self-esteem. These symptoms often severely disrupt patients' daily lives and professional performance.
[0003] Currently, the treatment of depression mainly involves a combination of medication and psychological intervention. In terms of medication, traditional antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), norepinephrine, and specific serotonergic antidepressants, alleviate depressive symptoms by regulating neurotransmitter function. However, while these medications are effective to some extent, they are accompanied by significant side effects, such as insomnia, nausea, and vomiting, and their effectiveness often takes a long time to materialize, posing challenges to patient experience and adherence.
[0004] Natural products have attracted increasing attention in the field of drug development due to their rich biological activity and relatively low side effects. Therefore, the search for and development of ideal antidepressants from natural medicines or traditional Chinese medicine formulas has gradually drawn researchers' attention.
[0005] Viola tianschanica Maxim., belonging to the Violaceae family and the Viola genus, is used medicinally as its dried whole plant, possessing heat-clearing and detoxifying properties. As a traditional medicinal plant, Viola tianschanica has a complex and diverse chemical composition, exhibiting various biological activities such as antioxidant, antibacterial, and anti-inflammatory effects. However, to date, specific research on the prevention or treatment of depression and related complications by Viola tianschanica or its extracts remains lacking. Therefore, further exploring the multiple effects of Viola tianschanica and promoting its expansion in practical medical applications has become an important research topic. Summary of the Invention
[0006] To address the significant side effects of traditional antidepressants, the present invention aims to provide the application of Viola yedoensis extract in the preparation of antidepressants.
[0007] To achieve the above objectives, the technical solution of the present invention is as follows.
[0008] This invention provides the application of Viola tianshanense extract in the preparation of antidepressant drugs, wherein the Viola tianshanense extract is an ethanol extract of Viola tianshanense dried powder.
[0009] This invention is the first to discover that Viola tianshanense extract possesses significant antidepressant activity, demonstrating its potential for preparing pharmaceutical formulations to prevent or treat depression or depression-related diseases, thus providing a new pharmaceutical basis for the development of antidepressant drugs. This invention verified the significant antidepressant activity of Viola tianshanense extract against depressive symptoms through sucrose preference tests, tail suspension tests, and forced swimming tests, addressing the problem of significant side effects associated with traditional antidepressant drugs. In this invention, Viola tianshanense extract contains multiple active ingredients that work synergistically to prevent or treat depression. Therefore, Viola tianshanense extract of this invention can be used as an active ingredient in the preparation of antidepressant drugs.
[0010] Preferably, the Viola tianshanense extract is obtained by extracting Viola tianshanense dried powder with ethanol; the mass percentage concentration of ethanol is 75% to 85%.
[0011] Preferably, the specific extraction method of the Viola tianshanense extract is as follows:
[0012] Viola tianshanense was used as raw material. After being crushed and sieved, Viola tianshanense dried powder was obtained. The Viola tianshanense dried powder was mixed with ethanol and then subjected to reflux reaction to obtain Viola tianshanense extract.
[0013] The extract of Viola tianshanensis was filtered, and the filtrate was subjected to vacuum distillation to remove ethanol, thus obtaining Viola tianshanensis extract.
[0014] Preferably, the mass ratio of dried Viola tianshanense powder to ethanol is 1:10-15.
[0015] Preferably, the concentration of the Viola tianshanense extract, calculated as crude drug, is 0.8 g / mL of Viola tianshanense dried powder. Specifically, each 1 mL of Viola tianshanense extract contains 0.8 g of Viola tianshanense dried powder.
[0016] Preferably, the extract yield of Viola tianshanense is 1:2-5. Specifically, the extract yield = mass of Viola tianshanense extract / mass of Viola tianshanense dried powder. That is, 2-5 parts by weight of Viola tianshanense dried powder can prepare 1 part by weight of Viola tianshanense extract.
[0017] Preferably, the reflux reaction time is 75 min to 105 min.
[0018] Preferably, the use of Viola tianshanense extract in the preparation of pharmaceutical formulations for the prevention or treatment of depression.
[0019] Preferably, the pharmaceutical preparation is an oral pharmaceutical preparation.
[0020] Preferably, the oral pharmaceutical preparation is a tablet, capsule, drop pill, sustained-release preparation, granule, or oral liquid preparation.
[0021] Preferably, the antidepressant also contains pharmaceutically acceptable excipients; the excipients include binders, fillers, diluents, lubricants, and flavoring agents.
[0022] Preferably, the excipient is at least one selected from starch, lactose, microcrystalline cellulose, dextrin, calcium phosphate, polyethylene glycol-4000, polyethylene glycol-6000, sodium carboxymethyl cellulose, hydroxypropyl cellulose, and crospovidone.
[0023] The beneficial effects of this invention are:
[0024] 1. This invention is the first to discover that Viola tianshanense extract has significant antidepressant activity and has the potential to prepare pharmaceutical formulations for the prevention or treatment of depression or depression-related diseases, providing a new pharmaceutical basis for the development of antidepressant drugs.
[0025] 2. The Viola tianshanensis extract of the present invention is mainly obtained by extracting Viola tianshanensis. Its raw materials are mainly from Xinjiang and other regions, which are rich in resources. Moreover, the preparation process of the extract is simple, low in cost, and suitable for large-scale production.
[0026] 3. The present invention verified through sugar water preference test, tail suspension test and forced swimming test that the extract of Viola tianshanensis of the present invention showed significant antidepressant activity in the treatment of depressive symptoms. Attached Figure Description
[0027] Figure 1 This is a graph showing the results of a sugar water preference test for the Viola tianshanense extract in this invention.
[0028] Figure 2 This is a diagram showing the results of the tail suspension test of the Viola tianshanense extract in this invention.
[0029] Figure 3 This is a diagram showing the results of the forced swimming test of the Viola tianshanense extract in this invention. Detailed Implementation
[0030] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.
[0031] Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.
[0032] Currently, the treatment of depression mainly involves a combination of medication and psychological intervention. In terms of medication, traditional antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), norepinephrine, and specific serotonergic antidepressants, alleviate depressive symptoms by regulating neurotransmitter function. However, while these medications are effective to some extent, they are accompanied by significant side effects, such as insomnia, nausea, and vomiting, and their effectiveness often takes a long time to materialize, posing challenges to patient experience and adherence.
[0033] Natural products have attracted increasing attention in the field of drug development due to their rich biological activity and relatively low side effects. Therefore, the search for and development of ideal antidepressants from natural medicines or traditional Chinese medicine formulas has gradually drawn researchers' attention.
[0034] Viola tianschanica Maxim., belonging to the Violaceae family and the Viola genus, is used medicinally as its dried whole plant, possessing heat-clearing and detoxifying properties. As a traditional medicinal plant, Viola tianschanica has a complex and diverse chemical composition, exhibiting various biological activities such as antioxidant, antibacterial, and anti-inflammatory effects. However, to date, specific research on the prevention or treatment of depression and related complications by Viola tianschanica or its extracts remains lacking. Therefore, further exploring the multiple effects of Viola tianschanica and promoting its expansion in practical medical applications has become an important research topic.
[0035] Unless otherwise specified, the methods described in the following embodiments are conventional methods; the reagents and materials mentioned are commercially available unless otherwise specified.
[0036] In the following embodiments, *Viola tianshanense* was purchased from the Hotan region medicinal herb market. The extract yield of the *Viola tianshanense* was 1:3-4. That is, 3-4 parts by weight of dried *Viola tianshanense* powder can prepare 1 part by weight of *Viola tianshanense* extract.
[0037] In the following examples, the mice used in the experiments were provided by the Laboratory Animal Institute of Lanzhou University.
[0038] Example 1
[0039] The preparation method of Viola tianshanense extract includes the following steps:
[0040] Weigh an appropriate amount of dried branches and leaves of Viola tianshanense, pulverize and sieve to obtain Viola tianshanense powder. Place the sieved Viola tianshanense powder into a round-bottom flask, add 75% (wt) ethanol at a material-to-liquid ratio of 1:15, reflux in a water bath for 90 min to obtain Viola tianshanense extract. Filter, combine the filtrates, and transfer the filtrate to a rotary evaporator at 45℃. Remove ethanol by vacuum distillation until a viscous concentrate is obtained, yielding Viola tianshanense extract. Based on crude drug content, each 1 mL of Viola tianshanense extract contains 0.8 g of Viola tianshanense powder.
[0041] Example 2
[0042] The preparation method of Viola tianshanense extract includes the following steps:
[0043] Weigh an appropriate amount of dried branches and leaves of Viola tianshanense, pulverize and sieve to obtain Viola tianshanense powder. Place the sieved Viola tianshanense powder into a round-bottom flask, add 85% (wt) ethanol at a material-to-liquid ratio of 1:10, reflux in a water bath for 105 min to obtain Viola tianshanense extract. Filter, combine the filtrates, and transfer the filtrate to a rotary evaporator at 45℃. Remove ethanol by vacuum distillation until a viscous concentrate is obtained, yielding Viola tianshanense extract. Based on crude drug quantity, each 1 mL of Viola tianshanense extract contains 0.8 g of Viola tianshanense powder.
[0044] Application Examples
[0045] Since the preparation methods of Examples 1 and 2 are basically the same, the yields of the *Viola tianshanense* extracts obtained are basically the same, and the concentrations of the *Viola tianshanense* extracts obtained in Examples 1 and 2 are basically the same. Therefore, the *Viola tianshanense* extracts used in the following animal experiments were prepared by the method described in Example 1.
[0046] The following application examples use the Viola tianshanensis extract prepared in Example 1 for antidepressant studies. The antidepressant activity of the Viola tianshanensis extract prepared in the embodiments of the present invention on depressive symptoms was verified by sucrose preference test, tail suspension test and forced swimming test.
[0047] 1. Establish a depression model.
[0048] Chronic unpredictable mild stress (CUMS) was used to induce depressive-like behavioral and physiological symptoms in mice. Mice were provided by the Laboratory Animal Institute of Lanzhou University. Mice were randomly subjected to a series of mild stresses for six consecutive weeks to mimic the pathophysiological characteristics of human depression.
[0049] The specific process is as follows:
[0050] Eight-week-old C57BL / 6 mice were used. In the initial stage of the experiment, the mice were allowed to acclimatize to the environment for one week, with free access to water and food, and a 12-hour light / dark cycle. The ambient temperature was 18–22℃, and the humidity was 50%–60%.
[0051] For the next six weeks, the following mild stressors were randomly applied, with one to two different stressors per day, and the same stressor was not repeated each week. The specific stressors included: wet housing, confined space, nighttime lighting, tilted cage, no water or no food, swimming in cold water, and empty cage.
[0052] Wet rearing involves placing moist bedding at the bottom of the mouse cage, maintaining a humidity level of 70%–90% for 24 hours.
[0053] Confined space involves placing mice in a narrow container to restrict their movement for 5 to 8 hours.
[0054] Nighttime lighting involved keeping the lights on continuously for 24 hours during the mice's nocturnal activity period.
[0055] Tilting the cage involves inverting or tilting the cage at a 45-degree angle for 24 hours.
[0056] Fasting or withholding water or food involves periodically removing food or water for 24 hours.
[0057] Cold water swimming involves using cold water at 0℃ to 4℃ and swimming for 2 to 3 minutes.
[0058] An empty cage is one without bedding material for 24 hours.
[0059] 2. Experimental methods for behavioral tests.
[0060] After the stress procedure is completed, the following behavioral tests are performed to assess depressive symptoms:
[0061] The sugar water preference test (SPT) primarily involves giving mice a choice between tap water and sugar water, measuring their preference for the sugar water to reflect the degree of their loss of pleasure. The specific experimental procedures are as follows:
[0062] One to two days before the experiment, the mice need to be allowed to acclimatize to the experimental environment and equipment. Place the mice in an experimental cage with two water bottles inside: one containing 5% sugar solution and the other containing plain water. Allow the mice free access to drink and observe their drinking behavior.
[0063] At the start of the experiment, the initial volumes of the two bottles of water were recorded. Mice were allowed free access to both types of water for 24 hours. The remaining volumes of water in both bottles were recorded after 6 hours and 12 hours. The volumes of sugar water and plain water consumed by the mice over 24 hours were calculated.
[0064] The formula for calculating the sugar water preference index is:
[0065] Sugar water preference index = Sugar water intake / (Sugar water intake + Water intake) × 100%.
[0066] The tail suspension test (TST) primarily records the time a mouse remains inactive while suspended upside down with its tail tied. Inactivity means giving up struggling. The specific experimental procedures are as follows:
[0067] Before conducting the experiment, it is necessary to ensure that the suspension is secure, the clamps or holding devices are in place, and the equipment for recording behavior is functioning properly.
[0068] Once the experiment begins, the mouse is suspended from a hanging frame with its tail clamped in a clip or holding device. The mouse's body cannot touch the ground or other supports, preventing it from escaping.
[0069] Behavioral recording devices were used to record the behavior of mice in a suspended state. The suspension adaptation time was 1-2 minutes, and the suspension test time was 4 minutes. The main focus was on whether the mice exhibited depressive behaviors such as ceasing to struggle, curling up in a ball, and becoming apathetic. The time the mice remained in the suspended state was recorded by software, usually in seconds. This parameter is considered an indicator of the degree of depressive behavior, and drug treatment may reduce this time.
[0070] Statistical methods were used to analyze the experimental results. The behavioral performance of mice in different groups under suspension was compared to evaluate the efficacy of the drug.
[0071] The forced swimming test (FST) primarily records the time a mouse remains afloat but inactive in water. The specific experimental procedures are as follows:
[0072] Before the experiment, the forced swimming tub was thoroughly cleaned to remove any animal odor. Specifically, an open cylindrical container with a total volume of 2500 mL, a height of 20 cm, and a diameter of 14 cm was filled with 10 cm of water at a temperature of 23℃ to 25℃.
[0073] Mice were placed inside a cylinder, and the forced swimming time was recorded for 6 minutes. The first 2 minutes were the adaptation phase for the mice to the new environment, and the last 4 minutes were the actual experimental testing phase.
[0074] During the experiment, the behavior of the mice needs to be recorded. Mouse behavior includes both active and immobile states. Active states include struggling, swimming, and other movements. Immobile states are characterized by the mouse giving up struggling, floating motionless in the water, and only making necessary movements to keep its head above water. The duration of immobility is an important indicator of the animal's level of depression. After the time is up, the mice are removed, dried, and the water in the container is changed and the mice are rinsed before the next mouse is tested.
[0075] The experimental results were analyzed using statistical analysis methods to compare the struggling time of mice in different groups.
[0076] 3. Experimental grouping.
[0077] The experiment was divided into the following four groups:
[0078] Control group: After being fed under normal conditions for 3 weeks, physiological saline was administered by gavage daily.
[0079] Model group: After the third week of modeling, physiological saline was administered by gavage.
[0080] Drug administration group: After the third week of modeling, the Viola tianshanense extract from Example 1 was dissolved in physiological saline solution to prepare a 20 mg / mL Viola tianshanense extract solution. The solution was administered by gavage at a dose of 400 mg / kg. Behavioral tests were conducted 3 weeks after administration.
[0081] Positive control group: Fluoxetine hydrochloride was administered by gavage at a dose of 40 mg / kg 3 weeks after modeling.
[0082] 4. Test results.
[0083] 4.1 Sugar water preference test.
[0084] The results of the sugar water preference test are as follows Figure 1 .
[0085] Depend on Figure 1 The results showed that, compared with the control group, the mice in the model group drank more pure water, a significant difference. In other words, the mice in the model group showed a significantly reduced preference for sucrose water, indicating the successful establishment of the depression model. Compared with the model group, the mice in the antidepressant treatment group showed a significant preference for sucrose water. This indicates that Viola tianshanense extract can significantly increase the sucrose water preference of the mice in the model group. Furthermore, Viola tianshanense extract itself did not affect the sucrose water preference of normal mice.
[0086] The above analysis shows that Viola tianshanense extract can significantly improve the depressive behavior of depressed mice, mainly by significantly increasing the sucrose preference of depressed model mice.
[0087] 4.2. Tail suspension test.
[0088] The results of the tail suspension test are as follows Figure 2 .
[0089] Depend on Figure 2The results showed that the model group had a longer immobility time (i.e., a longer resting time) compared to the control group, indicating a significant difference and successful establishment of the depression model. Compared to the model group, the depressant-treated group had a shorter immobility time (i.e., a shorter resting time), also showing a significant difference. This indicates that Viola tianshanense extract can significantly reduce the immobility time in the model group mice. Furthermore, Viola tianshanense extract itself did not affect the immobility time in normal mice.
[0090] The above analysis shows that Viola tianshanense extract can significantly improve depressive behavior in depressed mice, mainly by significantly reducing the hanging immobility time of the depressed model mice.
[0091] 4.3 Forced swimming test.
[0092] The results of the forced swimming test are as follows Figure 3 .
[0093] Depend on Figure 3 The results showed that the model group had a significantly longer immobile time in water compared to the control group, indicating a successful establishment of the depression model. The antidepressant treatment group had a significantly shorter immobile time compared to the model group. This demonstrates that Viola tianshanense extract can significantly reduce the immobile time in water for mice in the model group. Furthermore, Viola tianshanense extract itself did not affect the immobile time in water for normal mice.
[0094] The above analysis shows that Viola tianshanense extract can significantly improve depressive behavior in depressed mice, mainly by significantly reducing the immobility time in water in the depressed model mice.
[0095] In summary, the *Viola tianshanensis* extract provided in this invention significantly improves depressive-like behavior in mice induced by chronic unpredictable mild stress. This is mainly manifested in a significant increase in the percentage of sucrose preference in the sucrose preference test; a significant reduction in the immobility time in the tail suspension test; and a significant reduction in the immobility time in the water during the forced swimming test. Therefore, the *Viola tianshanensis* extract provided in this invention has the potential to prepare pharmaceutical formulations for the prevention or treatment of depression or depression-related diseases, providing a new pharmaceutical basis for the development of antidepressants and offering a treatment option with fewer side effects and faster onset of action.
[0096] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. The application of Viola tianshanense extract in the preparation of antidepressant drugs, characterized in that, The Viola tianshanense extract is an ethanol extract of Viola tianshanense dried powder; The specific extraction method of the Viola tianshanense extract is as follows: Viola tianshanense was used as raw material. After being crushed and sieved, Viola tianshanense dried powder was obtained. The Viola tianshanense dried powder was mixed with ethanol and subjected to reflux reaction to obtain Viola tianshanense extract. The Viola tianshanense extract was filtered, and the filtrate was subjected to vacuum distillation to remove ethanol to obtain Viola tianshanense extract. The mass percentage concentration of ethanol was 75% to 85%.
2. The application according to claim 1, characterized in that, The mass ratio of dried Viola tianshanense powder to ethanol is 1:10-15.
3. The application according to claim 1, characterized in that, The concentration of the Viola tianshanense extract, calculated as crude drug, is 0.8 g / mL containing dried Viola tianshanense powder.
4. The application according to claim 1, characterized in that, The extract yield of the Viola tianshanense was 1:2 to 5.
5. The application according to claim 1, characterized in that, The reflux reaction time is 75 min to 105 min.
6. The application according to claim 1, characterized in that, Application of Viola tianshanense extract in the preparation of pharmaceutical formulations for the treatment of depression.
7. The application according to claim 6, characterized in that, The pharmaceutical preparation is an oral pharmaceutical preparation.
8. The application according to claim 7, characterized in that, The pharmaceutical preparation also contains pharmaceutically acceptable excipients; the excipients include binders, fillers, diluents, lubricants, and flavoring agents.