Application of dihydroquercetin in preparation of antidepressant drug

Abstract

The invention provides an application of dihydroquercetin in preparation of an antidepressant drug. The dihydroquercetin is found to be capable of effectively improving the depressive symptom induced by IBD for the first time, and compared with a traditional treatment method, the medicine (TAX) shows a more excellent effect in the aspect of improving the depressive symptom of IBD mice. Through behavioral tests, such as a forced swimming test and an open-field test, the TAX obviously reduces the immobility time and the floating time of a mouse under a medium dose (50mg / kg), and shows an obvious anti-depression effect. Therefore, the dihydroquercetin has a great application prospect in preparation of the antidepressant drugs.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to the application of dihydroquercetin in the preparation of antidepressant drugs. Background Art

[0002] Depression is a form of bipolar disorder, characterized by typical symptoms such as low mood, slowed thinking, and reduced and slowed speech and motor activities. Depression seriously troubles patients' lives and work, bringing a heavy burden to families and society. Traditional antidepressant drugs have limitations. For example, monoamine inhibitors (SSRIs / SNRIs) represented by drugs: sertraline, fluoxetine, venlafaxine. Mechanism of action: selectively inhibit 5-HT / NE reuptake. Limitations: slow onset of action (2 - 6 weeks), large individual differences in efficacy; only relieve symptoms in 50% - 60% of patients (remission rate); unable to improve comorbid pathologies such as intestinal inflammation. There are also tricyclic antidepressants (TCAs) represented by drugs: amitriptyline, imipramine. Disadvantages: serious side effects (anticholinergic reactions, arrhythmias), and gradually replaced by SSRIs.

[0003] Therefore, it is necessary to develop an antidepressant drug with better pharmacological effects, multi-target regulation, higher safety than chemically synthesized drugs, and low side effect risks. Summary of the Invention

[0004] The object of the present invention is to provide the application of dihydroquercetin in the preparation of antidepressant drugs. The present invention discovers for the first time that dihydroquercetin can effectively improve the depressive symptoms induced by IBD, indicating that dihydroquercetin has great application prospects in the preparation of antidepressant drugs.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] In the first aspect of the present invention, there is provided the application of dihydroquercetin in the preparation of antidepressant drugs.

[0007] Furthermore, the drug further comprises pharmaceutically acceptable excipients and carriers.

[0008] Furthermore, the excipients include at least one of fillers, disintegrants, binders, excipients, lubricants, sweeteners, colorants.

[0009] Furthermore, the dosage form of the drug includes at least one of granules, tablets, pills, capsules, injections.

[0010] Furthermore, the effective dose of dihydroquercetin is 25 - 50 mg / kg.

[0011] In the second aspect of the present invention, there is provided an antidepressant drug, and the active ingredient of the drug includes dihydroquercetin.

[0012] In the third aspect of the present invention, a method for constructing an IBD-depression comorbidity animal model is provided, and the method includes the following steps:

[0013] First, perform animal grouping: 8-week-old C57BL / 6j mice are randomly divided into a control group, an IBD group, and a TAX group;

[0014] Through IBD induction, all groups drink 2.5% DSS for 7 days to construct an inflammatory bowel disease (IBD) animal model;

[0015] After the TAX group is administered dihydroquercetin, evaluate the depressive behavior phenotype.

[0016] Furthermore, the evaluation of the depressive behavior phenotype includes:

[0017] Behavioral science: Forced swimming and tail suspension experiments are used to evaluate depressive / anxiety behaviors;

[0018] Physiological indicators: Record body weight, fecal traits, and occult blood daily, calculate the DAI index, and serum neurotransmitter levels;

[0019] Pathological detection: HE staining is used to evaluate intestinal tissue inflammatory infiltration.

[0020] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0021] 1. The present invention first discovers that dihydroquercetin can effectively improve the depressive symptoms induced by IBD, indicating that dihydroquercetin has great application prospects in the preparation of antidepressant drugs.

[0022] 2. Dual improvement effect and comprehensive treatment effect: The present invention can not only improve the depressive behavior of IBD mice, but also effectively relieve the intestinal inflammatory response caused by IBD. The evaluation results of the DAI index show that the mice in the TAX treatment group are significantly lower in inflammatory indicators than the untreated group. Especially at the dose of 50 mg / kg, both depressive behavior and intestinal inflammation are improved. This dual effect enables the present invention to have strong comprehensive treatment ability and can act on both the intestinal symptoms of IBD and the depressive symptoms caused by it simultaneously.

[0023] 3. Dihydroquercetin has the advantages of natural compounds in the preparation of antidepressant drugs: It has higher safety than chemically synthesized drugs and lower side effect risks (such as hepatotoxicity). Existing preparation processes (such as tablets and capsules) have low industrialization costs.

[0024] 4. This compound is safe and effective, and no obvious toxic and side effects are found during all the experimental processes of the present invention. Dose optimization: The dose of 50 mg / kg has both high efficiency and safety (no significant hepatotoxicity)

[0025] 5. Structural simplification and convenient processing: The pharmaceutical preparation of the present invention is relatively simple, adopting conventional administration methods (oral or injection), and the production process of the drug is not complex. This not only reduces the production cost, but also improves the production efficiency, facilitating large-scale application. Compared with some drugs that require complex processes, the present invention has higher operability and a lower production threshold, and is easy to be industrially produced.

[0026] 6. Environmentally friendly and pollution reduction: In the drug preparation process of the present invention, attention is paid to reducing environmental pollution during the selection of raw materials and reaction processes, adopting green chemical technologies and environmentally friendly materials. This plays a positive role in reducing possible environmental pollution during drug production. At the same time, the treatment plan of the present invention can also indirectly reduce the burden on social resources by alleviating IBD and its depressive symptoms, reducing the pain of patients, and thus improving the quality of life of patients. Description of the Drawings

[0027] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0028] Figure 1 For the results of behavioral tests, where A: Forced swimming experiment; B: Tail suspension experiment; C: Change in immobility time of the forced swimming experiment in the behavioral study of the present invention; D: Change in immobility time of the tail suspension experiment in the behavioral study of the present invention;

[0029] Figure 2 For the results of colon length measurement, where A is the change in colon length of the mice of the present invention induced by DSS and the intervention group in the IBD of the present invention; B: Change in DAI index. Detailed Embodiments

[0030] The following will specifically describe the present invention in combination with specific embodiments and examples, and the advantages and various effects of the present invention will be presented more clearly therefrom. Those skilled in the art should understand that these specific embodiments and examples are used to illustrate the present invention, rather than to limit the present invention.

[0031] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as having the meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In case of contradiction, this specification shall prevail.

[0032] Unless otherwise specified, various raw materials, reagents, instruments, and equipment used in the present invention can be obtained through market purchases or by existing methods. Dihydroquercetin, CAS No.: 29838-67-3, can be commercially purchased.

[0033] The present application will be described in detail below in conjunction with examples and experimental data.

[0034] Example 1

[0035] I. Steps and methods:

[0036] 1. Animal selection and adaptive feeding:

[0037] Animal selection: Healthy 8-week-old C57BL / 6j mice were selected. This mouse strain is commonly used in experimental research and has strong adaptability.

[0038] Adaptive feeding: Before the experiment began, the mice were given one week of adaptive feeding time to ensure their health status and eating habits.

[0039] 2. Group division and modeling treatment:

[0040] Grouping: The mice were randomly divided into 4 groups:

[0041] Control group

[0042] IBD group (only induced IBD model)

[0043] Low-dose TAX administration group

[0044] Medium-dose TAX administration group

[0045] Modeling treatment: At the beginning of the experiment, all experimental groups (including the IBD group and the TAX administration group) were provided with a 2.5% dextran sulfate sodium (DSS) solution as drinking water daily for 7 consecutive days. DSS induces intestinal inflammatory responses by disrupting the intestinal barrier, simulating the occurrence of IBD.

[0046] Free diet: The mice maintained a free diet throughout the experiment.

[0047] 3. TAX administration:

[0048] TAX administration: Starting from the 8th day, TAX (dihydroquercetin) was administered:

[0049] Low-dose TAX group: Each mouse was gavaged with 0.5 ml of TAX solution daily at a dose of 25 mg / kg.

[0050] Medium-dose TAX group: Each mouse was gavaged with 1 ml of TAX solution daily at a dose of 50 mg / kg.

[0051] These administrations lasted for 7 days.

[0052] Control group and IBD group: These two groups did not receive TAX administration and had free access to food.

[0053] 4. Behavioral tests:

[0054] On the 7th day after the end of administration (i.e., the 15th day of the experiment), the following behavioral tests were conducted on all mice:

[0055] Forced swimming test (FST): Evaluate the immobility time of mice in water to reflect the severity of depressive symptoms.

[0056] Tail suspension test (TST): Evaluate the behavioral responses of mice due to stress, usually by measuring the immobility time of mice when they are suspended to assess the levels of anxiety and depression.

[0057] The key parameters of the forced swimming test (FST) and the tail suspension test (TST) are as follows:

[0058] The duration of the experiment is 5 minutes for FST and 5 minutes for TST;

[0059] The water temperature is controlled at 25 °C;

[0060] Only record the total immobility time at the observation time point.

[0061] Record the immobility time of each group of mice as an indicator of depressive behavior.

[0062] Body weight and feces monitoring:

[0063] Body weight monitoring: Record the body weight of mice daily to observe whether there is weight loss or other health changes.

[0064] Fecal characteristics and occult blood monitoring: Observe and record the fecal changes of mice, including the appearance of feces, loose stools, and whether there is an occult blood reaction. Based on this information, evaluate the severity of IBD.

[0065] DAI index: Based on weight loss, fecal characteristics, and occult blood reaction, calculate and analyze the DAI (disease activity index) to quantify the severity of IBD.

[0066] 5. Serum neurotransmitter analysis:

[0067] On the 7th day after the end of administration (i.e., the 15th day of the experiment), blood was taken from the fundus of the eyes of all mice, the collected serum was separated, and the levels of dopamine and 5-hydroxytryptamine were detected.

[0068] Record and analyze the changes in serum neurotransmitter levels in each group as an indicator of depression.

[0069] 6. Measurement of Colon Length

[0070] On the 7th day after the end of drug administration (i.e., the 15th day of the experiment), all mice were anesthetized and dissected. The colons of the mice were taken in vitro, and the colon lengths of the mice were measured as an index to determine whether the DSS-induced model was successful and whether TAX intervention had an improvement effect on inflammatory bowel disease (IBD).

[0071] II. Results

[0072] According to Figure 1 it is known that

[0073] Figure 1 A: The Forced Swimming Test is usually used to evaluate the depressive behavior of animals. In the experiment, the animal is placed in a container filled with water, and its swimming, struggling, pausing and other behaviors are observed within a certain period of time. By comparing the activity status of the animal in the experiment, its stress response and emotional state can be analyzed. Figure 1 Comparison of immobility time in the Forced Swimming Test (FST) in A

[0074] Control group: The immobility time was 8.2 ± 1.5 seconds (mean ± SD, n = 10);

[0075] IBD group: The immobility time was significantly prolonged to 24.6 ± 3.1 seconds (p < 0.01 vs control group);

[0076] Low-dose TAX group (25 mg / kg): The immobility time decreased to 18.3 ± 2.7 seconds (#p < 0.05 vs IBD group);

[0077] Medium-dose TAX group (50 mg / kg): The immobility time was further reduced to 12.1 ± 1.9 seconds (##p < 0.01 vs IBD group, no significant difference from the control group).

[0078] Figure 1 B: The Tail Suspension Test is an experimental method for evaluating depressive behavior. The tail of the animal is suspended and cannot rely on the ground for support. By observing the reaction of the animal in the suspended state, such as the immobility time, struggling behavior, etc., its emotional changes can be evaluated, and it is usually used to evaluate the effect of antidepressant drugs. According to the design of these two experimental methods, they can be used to evaluate the anxiety, depression and other emotional reactions of animals, and thus provide an experimental basis for studying the effect or behavioral mechanism of antidepressant drugs.

[0079] Figure 1 B: Comparison of immobility time in the Tail Suspension Test (TST)

[0080] Control group: The immobility time was 7.8 ± 1.3 seconds;

[0081] IBD group: The immobility time increased significantly to 23.4 ± 2.9 seconds (p < 0.01 vs control group);

[0082] Low-dose TAX group: The immobility time decreased to 17.9 ± 2.4 seconds (#p < 0.05 vs IBD group);

[0083] Medium-dose TAX group: The immobility time was significantly shortened to 11.5 ± 1.6 seconds (##p < 0.01 vs IBD group).

[0084] Figure 1 C: Compared with the control group, mice in the IBD group showed longer floating and less swimming, and the immobility time increased significantly; while in the TAX-administered groups, the immobility time increased compared with the control group but was lower than that in the IBD group, the floating time was shorter and there was occasional swimming; the medium-dose TAX group had a more significant reduction in immobility time compared with the low-dose TAX group, and stereotyped behavior decreased.

[0085] Figure 1 D: Compared with the control group, mice in the IBD group showed longer periods of stillness or gave up struggling, and the immobility time increased significantly; while in the TAX-administered groups, the immobility time increased compared with the control group but was lower than that in the IBD group, the stillness time was shorter and there was occasional struggling; the medium-dose TAX group had a more significant reduction in immobility time compared with the low-dose TAX group, and stereotyped behavior decreased.

[0086] According to Figure 2 it can be seen that:

[0087] Figure 2 A: Comparison of the colon lengths of mice in the control group and the IBD group. The average colon length of mice in the control group was 8 cm, while the average colon length of mice in the IBD group was significantly shortened to 5 cm, indicating that DSS-induced inflammation caused colon damage.

[0088] Figure 2 B: Compared with the control group, the DAI index of mice in the IBD group was significantly higher, indicating that DSS successfully induced an IBD model in mice and the inflammatory bowel disease activity in the modeled mice was stronger. The DAI index of the TAX-administered groups increased compared with the control group but was less than that in the IBD group; and the DAI index of the medium-dose TAX-administered group was lower than that of the low-dose TAX-administered group, indicating that the medium-dose TAX had a better remission effect on inflammatory bowel disease than the low-dose TAX.

[0089] The results of serum neurotransmitter detection are shown in Table 1. The results show that the levels of serum dopamine and 5-hydroxytryptamine in the IBD group of mice were significantly decreased. Low-dose TAX (25 mg / kg) could partially restore the neurotransmitter levels, but could not completely restore them to the control group level; medium-dose TAX (50 mg / kg) significantly increased the neurotransmitter levels, approaching the control group, showing a strong antidepressant effect. Statistical analysis showed that there was no significant difference between the medium-dose TAX group and the control group, suggesting that TAX has the potential to improve IBD-related depression.

[0090] Table 1. Serum neurotransmitter levels.

[0091]

[0092] Compared with the control group, *p < 0.05, **p < 0.01; compared with the IBD group, # p < 0.05, ## p < 0.01.

[0093] In summary, the present invention has the following advantages:

[0094] Multi-dimensional observation: In this experiment, through multiple dimensions such as behavioral tests, serum analysis, body weight monitoring, and fecal analysis, the improvement effect of TAX administration on the IBD-induced depression model was comprehensively observed.

[0095] Mechanism of action of TAX: Dihydroquercetin (TAX), as a natural anti-inflammatory and antioxidant compound, may have the potential to relieve IBD-related depression by regulating the immune response and the nervous system. This method helps to study the therapeutic effect of TAX in the co-morbidity of IBD and depression.

[0096] Combination of behavior and physiology: By combining behavioral tests (forced swimming test, tail suspension test) and physiological monitoring (body weight, fecal characteristics, DAI index, etc.), the comprehensiveness and reliability of the research results were ensured.

[0097] Finally, it should also be noted that the term "including", "comprising" or any other variation thereof is intended to cover non-exclusive inclusion, such that a process, method, article or device comprising a series of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article or device.

[0098] Although the preferred embodiments of the present invention have been described, additional changes and modifications can be made to these embodiments by those skilled in the art once they learn of the basic creative concept. Therefore, the appended claims are intended to be construed to include the preferred embodiments as well as all changes and modifications that fall within the scope of the present invention. Obviously, those skilled in the art can make various changes and variations to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention is also intended to include these modifications and variations.

Claims

1. Application of dihydroquercetin in the preparation of antidepressant drugs.

2. The use according to claim 1, characterized in that: The medicine also includes pharmaceutically acceptable excipients and carriers.

3. The use according to claim 1, characterized in that: The auxiliary materials include at least one of a filler, a disintegrant, a binder, a lubricant, a sweetener, and a colorant.

4. The use according to claim 1, characterized in that: The dosage form of the drug includes at least one of granules, tablets, pills, capsules and injections.

5. The use according to claim 1, characterized in that: The effective dosage of the dihydroquercetin is 25 mg / kg to 50 mg / kg.

6. The use according to claim 1, characterized in that: The dihydroquercetin reduces intestinal inflammation and improves neurological symptoms.

7. An antidepressant drug, characterized in that: The active ingredient of the drug includes dihydroquercetin.

8. An antidepressant drug according to claim 7, characterized in that: The drug also includes pharmaceutically acceptable excipients.

9. A method for constructing an IBD-depression comorbidity animal model, characterized in that: The method comprises the following steps: Animals were first grouped: 8-week-old C57BL / 6j mice were randomly divided into control group, IBD group, and TAX group; Through IBD induction, all groups drank 2.5% DSS for 7 days to construct an inflammatory bowel disease (IBD) animal model; After administration of dihydroquercetin, the TAX group was evaluated for depressive behavioral phenotype.

10. The method for constructing an IBD-depression comorbidity animal model according to claim 9, characterized in that: The evaluation of depressive behavioral phenotypes includes: Behavior: forced swimming and tail suspension tests to assess depression or anxiety behavior; Physiological indicators: daily record of body weight, stool characteristics and occult blood, calculation of DAI index, serum neurotransmitter levels; Pathological examination: HE staining was used to evaluate the inflammatory infiltration of intestinal tissue.

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