Application of geniposidic acid in the preparation of drugs for preventing or treating depression

Abstract

The present invention belongs to the field of biomedical technology and relates to the application of geniposidic acid in the preparation of drugs for preventing or treating depression. The geniposidic acid described in the present invention can be applied in the preparation of drugs for preventing or treating depression, and the therapeutic effect of geniposidic acid can be comparable to that of the commonly used existing antidepressant drug fluoxetine. Geniposidic acid can effectively reduce the immobility time in the tail suspension test; it can effectively reduce the immobility time in the forced swimming test; in the sucrose preference test, the high-dose geniposidic acid is equivalent to the positive control drug fluoxetine, indicating that geniposidic acid (5 mg / kg) alleviates the anhedonic behavior of CUMS mice; there are significant changes in the open field behavior indexes of the fluoxetine group and each dose group of geniposidic acid, among which the effect of the geniposidic acid (5 mg / kg) group is the same as that of fluoxetine (10 mg / kg). Geniposidic acid (5 mg / kg) can effectively reduce the levels of inflammatory factors IL-1β, TNF-α, and IL-6 in the mouse brain tissue. It can be known from the research data analysis that the therapeutic effect of geniposidic acid in treating depression can be comparable to that of the commonly used existing antidepressant drug fluoxetine.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine and relates to the application of geniposidic acid in the preparation of drugs for preventing or treating depression. Background Art

[0002] Depression is a common and serious mental illness, and its clinical manifestations include emotional depression, taciturnity, slow thinking, sleep disorders, physical pain. Self-harm and suicide are the most serious consequences of depression. [1] In 2008, major depressive disorder was ranked as the third largest disease in the global disease burden. By 2020, depression has become the second largest disease in the world. [3] It is predicted that this disease will rank first by 2030. [2] At present, worldwide, depression has become one of the diseases with the most serious burden on patients. [4] Currently, the first-line clinical treatment drugs include selective serotonin reuptake inhibitors (SSRIs), serotonin and norepinephrine reuptake inhibitors (SNRIs), bupropion, mirtazapine, and vortioxetine. Although these drugs play an important role in treating and improving the symptoms of depression, about one-third of depression patients do not significantly benefit from drug treatment and show a high recurrence rate and various side effects, such as nausea, headache, drowsiness, dry mouth, and male sexual dysfunction. [5] Therefore, searching for effective and safe antidepressants from Chinese herbal medicines has become an important research direction at home and abroad in recent years.

[0003] Current research results show that the pathogenesis of depression is mainly closely related to the mutual promotion of monoamine neurotransmitters and their receptors, inflammatory responses, hypothalamic-pituitary-adrenal (HPA) axis hormones, and neurotrophic factors (NTFs) in the body. [6-8] A large number of studies have shown that the contents of hypothalamic monoamine neurotransmitters such as 5-HT and its receptors, NA, DA, etc. in depressed patients are significantly reduced, activating the body's stress response and increasing the level of oxidative stress. Under the action of stress, the functions of the sympathetic nervous system and the HPA axis are more active, and then regulate the peripheral inflammatory response. The activated inflammatory factors will invade the central nervous system, leading to nervous system dysfunction and ultimately aggravating depression. [9-10] Thus, it can be seen that intervening in the inflammatory response can be used as one of the targets for treating depression.

[0004] Gardenia is a classic traditional Chinese medicine. Modern clinical pharmacological studies have shown that Gardenia has pharmacological effects such as protecting the liver and gallbladder, promoting pancreatic secretion, lowering blood sugar, lowering blood lipids, improving cerebral ischemia injury, and anti-inflammation. Gardenoside (Gar) is an effective monomer component in the Chinese herbal medicine Gardenia. Gardenia is the dried ripe fruit of the plant Gardenia jasminoides Ellis of the Rubiaceae family, which was first recorded in "Shennong Ben Cao Jing". However, so far, the therapeutic effect and application of Gardenoside in the treatment of depression have not been reported at home and abroad. The structural formula of Gardenoside is as follows:

[0005]

[0006] References:

[0007] [1] Yuan Y Y, Min H S, Lapane K L, et al. Depression symptoms and cognitive impairment in older nursing home residents in the USA: A latent class analysis[J]. Int J Geriatr Psychiatry, 2020, 35(7): 769 - 778.

[0008] [2] Chi X, Wang S, Baloch Z, et al. Research progress on classical traditional Chinese medicine formula Lily Bulb and Rehmannia Decoction in the treatment of depression[J]. Biomed Pharmacother. 2019 Apr; 112: 108616.

[0009] [3] Wang Y, Li M, Liang Y, et al. Chinese Herbal Medicine for the Treatment of Depression: Applications, Efficacies and Mechanisms[J]. Curr PharmDes. 2017; 23(34): 5180 - 5190.

[0010] [4]Haroz E.E., Ritchey M., Bass J.K., Kohrt B.A., et al. How is depression experienced around the world? A systematic review of qualitative literature[J]. Soc. Sci. Med. 2017; 183: 151 - 162.

[0011] [5]Kennedy SH, Lam RW, McIntyre RS, et al. Canadian Network for Mood and Anxiety Treatments (CANMAT) 2016 Clinical Guidelines for the Management of Adults with Major Depressive Disorder: Section 3. Pharmacological Treatments[J]. Can J Psychiatry 2016; 61: 540 - 60.

[0012] [6]Kim J W, Szigethy E M, Melhem N M, et al. Inflammatory markers and the pathogenesis of pediatric depression and suicide: A systematic review of the literature[J]. J Clin Psychiatry, 2014, 75(11): 1242 - 1253.

[0013] [7] Mu Lei, Sun Jianxu. Pathogenesis of depression and antidepressant drug targets[J]. International Journal of Pharmaceutical Research, 2015, 42(4): 463 - 466.

[0014] [8] Lamers F, Vogelzangs N, Merikangas K R, et al. Evidence for a differential role of HPA - axis function, inflammation and metabolic syndrome in melancholic versus atypical depression[J]. Mol Psychiatry, 2013, 18(6): 692 - 699.

[0015] [9] Wang Long, Wang Shitao. Research Progress on the Pathogenesis of Oxidative Stress in Depression and Acupuncture Treatment [J]. Journal of Clinical Acupuncture and Moxibustion, 2017, 33(11): 76-80.

[0016]

[10] Liu Zhiyang, Su Wenjun, Yan Wenjie, et al. Research Progress on the Inflammatory Mechanism of Depression [J]. Clinical Journal of Medical Officers, 2020, 48(12): 1513-1516. Summary of the Invention

[0017] The purpose of the present invention is to provide an application of geniposidic acid in the preparation of drugs for preventing or treating depression.

[0018] The implementation process of the present invention is as follows:

[0019] Application of geniposidic acid in the preparation of drugs for preventing or treating depression.

[0020] Furthermore, geniposidic acid can reduce the levels of inflammatory factors IL-1β, TNF-α, and IL-6 in the brain tissue of mice.

[0021] Furthermore, geniposidic acid can reduce the immobile time of mice in the tail suspension test.

[0022] Furthermore, geniposidic acid can reduce the immobile time of mice in the forced swimming test.

[0023] Furthermore, geniposidic acid can alleviate the anhedonic behavior of mice.

[0024] Furthermore, geniposidic acid can increase the exploratory behavior and spontaneous activity of mice in a new environment and reduce tension.

[0025] Furthermore, the drug for preventing or treating depression contains at least geniposidic acid.

[0026] Furthermore, the drug for preventing or treating depression includes geniposidic acid and a drug carrier.

[0027] Furthermore, the drug for preventing or treating depression is a tablet, capsule, pill, injection, sustained-release preparation, controlled-release preparation, or various particulate drug delivery systems.

[0028] Positive effects of the present invention:

[0029] (1) Geniposidic acid can be applied in the preparation of drugs for preventing or treating depression, and the therapeutic effect of geniposidic acid is comparable to that of the commonly used antidepressant fluoxetine.

[0030] (2) Geniposidic acid can effectively reduce the immobility time in the tail suspension test; it can effectively reduce the immobility time in the forced swimming test; in the sucrose preference test, the high-dose geniposidic acid is as effective as the positive control drug fluoxetine, indicating that geniposidic acid (5 mg / kg) alleviates anhedonic behavior in CUMS mice; significant changes have occurred in the open field behavior indices of the fluoxetine group and each dose group of geniposidic acid, among which the effect of the geniposidic acid (5 mg / kg) group is the same as that of fluoxetine (10 mg / kg).

[0031] (3) Geniposidic acid (5 mg / kg) can effectively reduce the levels of inflammatory factors IL-1β, TNF-α, and IL-6 in the mouse brain tissue. Description of the Drawings

[0032] Figure 1 It is a graph of the results of the mouse tail suspension test;

[0033] Figure 2 It is a graph of the results of the mouse forced swimming test;

[0034] Figure 3 It is a graph of the results of the mouse sucrose preference test;

[0035] Figure 4 It is a graph of the results of the mouse open field test, where A is the route map of the open field for each group of mice; B is the comparative analysis of the total distance of each group of mice; C is the comparative analysis of the central distance of each group of mice, and D is the comparative analysis of the central time of each group of mice;

[0036] Figure 5 It is a graph of the experimental results of detecting inflammatory factors by ELISA method. Detailed Embodiments

[0037] The present invention will be further described below in combination with experimental research.

[0038] The application of geniposidic acid described in the present invention in the preparation of drugs for preventing or treating depression. The drug for preventing or treating depression contains at least geniposidic acid, or includes geniposidic acid and a drug carrier. The drug for preventing or treating depression is a tablet, capsule, pill, injection, sustained-release preparation, controlled-release preparation, or various particulate drug delivery systems. Among them, the active ingredient geniposidic acid can reduce the levels of inflammatory factors IL-1β, TNF-α, and IL-6 in the mouse brain tissue, can reduce the immobility time of mice in the tail suspension test, can reduce the immobility time of mice in the forced swimming test, can alleviate the anhedonic behavior of mice, can increase the exploratory behavior and spontaneous activity of mice in a new environment, and reduce the tension.

[0039] Study on the antidepressant effect and mechanism of geniposidic acid

[0040] 1. Experimental animals

[0041] Male C57BL / 6 mice, weighing 22-25 g, were purchased from the Animal Center of Air Force Medical University (certificate number: 0009167). The breeding conditions were a temperature of 25±1°C and a relative humidity of 52±5%. They were adaptively bred for one week before the experiment.

[0042] 2. Drug preparation

[0043] Both gardoside and fluoxetine were dissolved in pure water.

[0044] The administration concentration of gardoside (Gar): 2.5 mg / kg, 5 mg / kg

[0045] The administration concentration of fluoxetine (Flu): 10 mg / kg

[0046] 3. Animal grouping

[0047] The experimental animals were randomly divided into a blank control group (Control), a model group (CUMS), a positive drug control group (CUMS+Flu), a low-dose gardoside group (CUMS+Gar-L), and a high-dose gardoside group (CUMS+Gar-H), with 10 mice in each group.

[0048] 4. Establishment of the CUMS model

[0049] A classical mouse CUMS depression model was established. The modeling methods included fasting for 24 h; water deprivation for 24 h; tilting the mouse cage at 45° for 24 h; using wet bedding (200 mL of water was added to 100 g of bedding) for 24 h; tail clamping for 1 min; cold water swimming at 4°C for 5 min; and circadian rhythm inversion for 24 h.

[0050] To prevent the mice from getting used to the occurrence of stress, all stressors were applied in a random and unpredictable order, and the same stimulus was not repeated within 24 hours, lasting for 6 weeks. The mice in the blank control group were placed in a mouse cage in a separate room and not given any stress. After 6 weeks of modeling, the drugs were administered at the above doses for 14 days.

[0051] 5. Behavioral experiments

[0052] 5.1 Tail suspension test (TST)

[0053] The mouse was suspended at a position 50 cm above the ground by sticking the tape about 1 cm from the tip of the mouse's tail. Each mouse was suspended for 6 min, with the first 2 min being the adaptation time, and the cumulative immobile time in the last 4 min was recorded. The criteria for judging the mouse's immobility were: the mouse's limbs were stationary or there were slight limb tremors.

[0054] 5.2 Forced swimming test (FST)

[0055] Mice were placed in a transparent cylinder with a height of 45 cm and a diameter of 20 cm, filled with 15 cm of water at a temperature of 24 ± 1 °C. Each mouse swam for 6 min, with the first 2 min being the adaptation time, and the cumulative immobile time in the subsequent 4 min was recorded. The criterion for determining that a mouse was immobile was that the mouse floated motionless on the water surface, stopped struggling, and had only minor limb movements.

[0056] 5.3 Sucrose preference test (SPT)

[0057] Seventy-two hours before the test, two bottles of 1% sucrose solution were provided simultaneously. Forty-eight hours before the test, one of the 1% sucrose solution bottles was replaced with pure water for adaptation. Twenty-four hours before the test, food and water were withheld. The specific test method was as follows: Mice were housed individually. After 24 h of food and water deprivation, each mouse was given two bottles of drinking water with exactly the same appearance (one bottle was 1% sucrose solution and the other was tap water). Both bottles of drinking water were weighed before placement, and it was ensured that the water outlet did not leak automatically. After 1 h, the weight of the bottles was measured again to determine the amount of sucrose solution or water consumed. The sucrose preference was calculated according to the following formula: Sucrose preference (%) = sucrose solution consumption / (sucrose solution consumption + tap water consumption) × 100%.

[0058] 5.4 Open field test (OFT)

[0059] Each mouse in each group was placed individually in an open field box (size 100 × 100 × 40 cm, with the inner wall painted black with dye). The bottom surface was evenly divided into 25 (5 × 5) squares, including 9 central squares in the center and the remaining 16 peripheral squares along the side walls. During the experiment, the mouse was placed in the center of the bottom surface of the box. The mouse was allowed to move freely for 5 min, and the number of squares it crossed within 5 min was recorded. After each mouse was tested, the inside of the open field box was cleaned to avoid affecting the next test.

[0060] 6. Research results of geniposide on the reversible mouse depressive behaviors induced by CUMS

[0061] 6.1 Effect of geniposide on the immobility time in the tail suspension test

[0062] The results of the mouse tail suspension test showed that compared with the Control group, the immobility time of the mice in the depressive model group was significantly increased (P < 0.01); compared with the model group, fluoxetine (10 mg / kg) and geniposide (5 mg / kg) could effectively reduce the immobility time (P < 0.05); the low dose of geniposide (2.5 mg / kg) had no significant change. The results are shown in Table 1 and Figure 1 Results of the mouse tail suspension test ( n = 10).

[0063] Table 1. Effect of geniposide on the immobility time in the mouse tail suspension test ( n = 10)

[0064]

[0065] Note: Compared with the Control group, ## P < 0.01; compared with the CUMS group, *P < 0.05.

[0066] 6.2 Effects of geniposide on the immobility time in the forced swimming test The results of the forced swimming test showed that compared with the Control group, the immobility time of mice in the depression model group increased significantly (P < 0.01); compared with the model group, fluoxetine (10 mg / kg) and geniposide (5 mg / kg) could effectively reduce the immobility time (P < 0.05); the change in the immobility time of the low-dose geniposide (2.5 mg / kg) was not obvious. The results are shown in Table 2 and Figure 2 Results of the forced swimming test in mice ( n = 10), compared with the Control group, ## P < 0.01; compared with the CUMS group, * P < 0.05.

[0067] Table 2. Effects of geniposide on the immobility time in the tail suspension test in mice ( n = 10)

[0068]

[0069]

[0070] Note: Compared with the Control group, **P < 0.01; compared with the CUMS group, *P < 0.05.

[0071] 6.3 Effects of geniposide on sucrose preference

[0072] As Figure 3 shown, the results of the sugar water preference test showed that compared with the Control group, the sugar water preference rate of mice in the CUMS model group decreased significantly (p < 0.01), indicating that the mice in the depression model group had anhedonic behavior. Compared with the model group, the sugar water preference rates of fluoxetine (10 mg / kg) and geniposide (5 mg / kg) in mice increased significantly (p < 0.05 or p < 0.01), and the high-dose geniposide had the same effect as the positive control drug fluoxetine, indicating that geniposide (5 mg / kg) alleviated the anhedonic behavior of CUMS mice. The results are shown in Figure 3 Results of the sugar water preference test in mice ( n = 10), compared with the Control group, ## P < 0.01; compared with the CUMS group,* P < 0.05.

[0073] 6.4 Effect of geniposide on open field test

[0074] The results of the open field test showed that compared with the Control group, the total distance, the proportion of the central distance, and the proportion of the central time of the mice in the CUMS model group were significantly reduced (p < 0.01), indicating that the exploratory behavior and spontaneous activity of the mice in the depression model group were significantly reduced in the new environment, while the tension was significantly increased. Compared with the CUMS model group, the open field behavior indexes of the fluoxetine group and each dose group of geniposide were significantly changed, and the effect of the geniposide (5 mg / kg) group was the same as that of fluoxetine (10 mg / kg). The results are shown in Table 3 and Figure 4 Results of the open field test of mice ( n = 10), * P < 0.05, ** P < 0.01, where A is the mine field route map of each group of mice; B is the comparative analysis of the total distance of each group of mice; C is the comparative analysis of the central distance of each group of mice, and D is the comparative analysis of the central time of each group of mice.

[0075] Table 3. Effect of geniposide on the immobility time of mice in the tail suspension test ( n = 10)

[0076]

[0077]

[0078] Note: Compared with the Control group, **P < 0.01; compared with the CUMS group, *P < 0.05.

[0079] 7. Effect of geniposide on the levels of inflammatory factors in the brain tissue

[0080] The levels of inflammatory factors were detected by ELISA method. Specifically, an ELISA kit was used to detect the levels of inflammatory factors IL-1β, TNF-α, and IL-6 in the brain tissue of mice according to the instructions of the kit.

[0081] The results of the ELISA method showed that compared with the Control group, the levels of inflammatory factors TNF-α, IL-1β, and IL-6 in the brain tissue of the mice in the CUMS model group were significantly increased. After drug intervention, the levels of inflammatory factors could be reduced, especially the effects of fluoxetine (10 mg / kg) and geniposide (5 mg / kg) were better. The results are shown in Figure 5 , the experimental results of detecting inflammatory factors by ELISA method ( n = 10) Compared with the Control group, ## P < 0.01; compared with the CUMS group, *P < 0.05 or ** P < 0.01.

[0082] In summary, the geniposide of the present invention can be applied to the preparation of drugs for preventing or treating depression. From the analysis of research data, it can be seen that the therapeutic effect of geniposide is comparable to that of the commonly used antidepressant fluoxetine.

[0083] The above content is a further detailed description of the present invention in combination with specific preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For those of ordinary skill in the art to which the present invention pertains, without departing from the concept of the present invention, several simple deductions or substitutions can be made, which should all be regarded as belonging to the protection scope of the present invention.

Claims

1. Use of geniposidic acid as the only active ingredient in the preparation of a drug for preventing or treating depression, Characterized in that: Geniposidic acid can reduce the levels of inflammatory factors IL-1β, TNF-α, and IL-6 in the brain tissue of mice.

2. The use according to claim 1, Characterized in that: Geniposidic acid can reduce the immobile time of mice in the tail suspension test.

3. The use according to claim 1, Characterized in that: Geniposidic acid can reduce the immobile time of mice in the forced swimming test.

4. The use according to claim 1, Characterized in that: Geniposidic acid can relieve anhedonic behavior in mice.

5. The use according to claim 1, Characterized in that: Geniposidic acid can increase exploratory behavior and spontaneous activity of mice in a new environment and reduce tension.

6. The use according to claim 1, Characterized in that: The drug for preventing or treating depression contains a pharmaceutically acceptable carrier.

7. The use according to claim 1, Characterized in that: The drug for preventing or treating depression is a tablet, capsule, pill, injection, sustained-release preparation, controlled-release preparation or various particulate drug delivery systems.

Images