CN103816171B - New lactuside B purpose - Google Patents

Abstract

The invention provides a new application of Lactucoside B, specifically related to the application in the preparation of drugs for preventing and treating depression. The advantage of the present invention is that the pharmacological effect of Lactucoside B is obvious, mainly as follows: it can increase the weight of depression mice, increase the number of activities, prolong the swimming time of depression mice, shorten the immobility time of their swimming, and improve depression. Predominance of sucrose in mice reduces mortality in depressed mice. The invention provides a new drug for preventing and treating depression and brings good news to patients with depression.

Description

New application of lettucoside B

technical field

The present invention relates to the new application of Lactucoside B, in particular to the application of Lactucoside B in the preparation of medicaments for preventing and treating depression.

Background technique

Pterocypsela elata (Hemsl.) Shih in Act. Also known as high lettuce Lactuca elata Hemsl. , which is a plant of the genus Pteraceae in the family Asteraceae, is distributed in more than a dozen provinces including Jilin, Gansu, and Henan in my country. It has the effects of clearing heat and detoxifying, promoting blood circulation and removing blood stasis, and expelling wind. Lactuside B (lactuside B) is the main extract from the root of lettuce. Its molecular formula is C ₂₁ H ₃₂ O ₉ , and its general structure is as follows:

Among them, R is preferably Glc or H; R' is preferably CH ₂ OH or CHO.

It was early discovered that lettuceside B monomer has anti-cerebral ischemia and memory-improving effects.

Depression is a common mood disorder. The main symptoms include significant and persistent low mood or loss of interest, often accompanied by sleep disturbance, excessive worry, low self-evaluation, social avoidance, and the patient feels extremely painful subjectively. Depression is characterized by high rates of suicide, self-harm, and relapse. As the incidence of mental illness increases year by year, depression has become the primary threat to the health of residents in our country, seriously affecting people's quality of life. It is estimated that the number of patients with depression in my country has reached 26 million, and the average incidence rate is about 2%. Most cases have recurrent attacks, and some may have residual symptoms or become chronic. Most patients need long-term medication. Therefore, effective prevention and treatment Treating depression is very important.

The etiology of depression is not very clear, the current research mainly focuses on genetic factors, neurobiochemical changes, abnormal endocrine function, neuroimaging changes and psychosocial factors. However, there is still no systematic theory that can explain well the internal connection between these factors and how to form the pathological mechanism of the occurrence and development of depression. At present, there are more studies on the dysfunction of neurotransmitter pathways in patients with depression, such as the abnormal secretion of serotonin and norepinephrine. Studies have found that the depletion of serotonin and norepinephrine can easily induce emotional changes such as depression, anxiety, and fear, but the specific mechanism is still unclear.

There is no report on the antidepressant effect of Lactucoside B.

Contents of the invention

The purpose of the present invention is to provide a new application of raptoside B. the

The invention provides the application of Lactucoside B in the preparation of medicaments for preventing and treating depression.

In the application provided by the present invention, the proposed clinical dosage of Lactucoside B is: 100-400mg/d.

The medicine is a preparation containing lettucoside B.

The preparation is made from raptucoside B alone or from raptucoside B and a pharmaceutically acceptable carrier.

The preparation is tablet, capsule, pill, oral liquid, granule or injection, and the tablet is common tablet or sublingual tablet.

The pharmaceutically acceptable carrier refers to a conventional drug carrier in the pharmaceutical field, selected from fillers, binders, disintegrants, lubricants, solubilizers, suspending agents, wetting agents, pigments, essences, solvents, One or more of surfactants or flavoring agents.

The filler is selected from starch, pregelatinized starch, dextrin, glucose, sucrose, lactose, lactitol, microcrystalline cellulose, mannitol, sorbitol or xylitol, preferably sorbitol, microcrystalline cellulose, lactose or pregelatinized starch;

The disintegrating agent is selected from cross-linked carmellose sodium, cross-linked povidone, low-substituted hydroxypropyl cellulose, sodium carboxymethyl starch or starch, preferably cross-linked povidone, low-substituted hydroxypropyl Cellulose or sodium carboxymethyl starch;

Described lubricant is selected from magnesium stearate, talcum powder, micropowder silica gel, PEG4000, PEG6000 or sodium lauryl sulfate, preferably magnesium stearate, talcum powder;

The binder is selected from sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, povidone, starch slurry, sucrose, powdered sugar, mucilage, gelatin or polyethylene glycol, preferably hydroxyl Propylmethylcellulose, povidone;

The solubilizer is selected from sodium hydroxide, potassium hydroxide, sodium bicarbonate, meglumine, L-lysine or L-arginine, preferably sodium hydroxide, meglumine;

The suspending agent is selected from micropowder silica gel, beeswax, cellulose or solid polyethylene glycol;

The wetting agent is selected from glycerin, Tween-80, ethoxylated hydrogenated castor oil or lecithin;

The solvent is selected from ethanol, liquid polyethylene glycol, isopropanol, Tween-80, glycerin, propylene glycol or vegetable oil, and the vegetable oil is selected from soybean oil, castor oil, peanut oil, blended oil, etc.;

The surfactant is selected from sodium dodecylbenzenesulfonate, stearic acid, polyoxyethylene-polyoxypropylene copolymer, fatty acid sorbitan or polysorbate (Tween), etc.;

The flavoring agent is selected from aspartame, sucralose, essence, stevia, acesulfame potassium, citric acid or sodium saccharin.

The new application of the lettucoside provided by the invention has the following advantages:

1. The present invention provides a new application of Lactucoside B, which is used for the prevention and treatment of depression, and brings new drug options for patients with depression;

2. The drug has remarkable effect and no side effects;

3. Good water solubility, easy to transform into clinical drugs, and has good application prospects;

4. The experimental results suggest that: Lactucoside B can increase the weight of depressed mice, increase their exercise performance, prolong their swimming time, and improve their preference for sucrose water. The above effects are dose-dependent, and their effects are related to Fluoxetine, an antidepressant commonly used in clinical practice, has similar effects, suggesting that Lactucoside B has a therapeutic effect on depression; preventive use of Lactucoside B can significantly reduce the mortality of depressed mice, increase their body weight and exercise performance, and prolong animal life. Swimming time, suggesting that lettucoside B has a better preventive effect on depression.

The results show that: Lactuca glycoside B has obvious preventive and therapeutic effects on mental depression induced by various reasons.

Detailed ways

The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

Example 1: Therapeutic Effect of Lactucoside B on Depression in Mice

1. Experimental animals: 80 male Kunming mice, weighing 20-22 g, were purchased from the Experimental Animal Center of Zhengzhou University.

2. Experimental reagents and instruments:

Lactucoside B (prepared by Xinxiang Medical College School of Pharmacy, purity greater than 95%), fluoxetine hydrochloride (Eli Lilly Suzhou Pharmaceutical Co., Ltd.), swimming pool, Open·Field open box.

3. Experimental grouping and experimental methods:

Before the experiment, 80 mice were adaptively fed for 1 week with 12 hours of light and 12 hours of no light, and then 10 mice were randomly selected as the normal control group: they were divided into two cages and raised in groups, with 5 mice in each cage. Feed and drinking water were given normally according to the needs, and no stimulation was accepted. The remaining mice were modeled according to the following method. After 21 days, 50 mice successfully modeled were selected and randomly divided into 5 groups, 10 mice in each group.

Mice were housed in a single cage and received seven different chronic stress stimuli at the same time. Including: overnight lighting deprivation of sleep environment, fasting (24h), water deprivation (24h), ice water swimming (4°C, 5min), hot water swimming (40°C, 5min), tail pinch (2min), tail suspension (3min) ). One kind of stimulus was randomly arranged every day, and the same kind of stimulus appeared discontinuously. In the 28 days of the experiment, each kind of stimulus was used 4 times on average, so that the mice could not predict the upcoming stimulus. Mice that were successfully modeled at 21 days after stimulation were divided into the following 5 groups:

Depression group (model group): within 28 days of the experiment, the mice in this group were fed in single cages and received various chronic stress stimuli. After 21 days, 0.1ml/10g of normal saline was injected intraperitoneally at 8:00 a.m. every day. Injected for 7 days.

Fluoxetine hydrochloride group (positive control group): the feeding environment and stimulation methods were the same as those of the depression group, and 0.5ml of the drug dissolved in double distilled water was administered orally at 8:00 a.m. every day at 35 μg/10g for 7 consecutive days;

Lactucoside B low-dose group: the feeding environment and stimulation methods were the same as those of the depression group. After 21 days, at 8:00 a.m., 12.5 μg/10 g intraperitoneally injected 0.5 ml of Lactucoside B dissolved in double distilled water for 7 consecutive days;

Rapunzel B medium dose group: the feeding environment and stimulation methods were the same as those of the depression group. After 21 days, at 8:00 am every day, 25 μg/10 g intraperitoneally injected 0.5 ml of lettucoin B dissolved in sterilized double distilled water for 7 consecutive days;

Lactucoside B high-dose group: The feeding environment and stimulation methods were the same as those of the depression group. After 21 days, 0.5ml of the drug dissolved in sterilized double-distilled water was injected intraperitoneally at 8:00 a.m. at 50μg/10g for 7 consecutive days.

The above-mentioned fluoxetine hydrochloride group and the low-, middle-, and high-dose groups of lettucoside B still continued to stimulate during the administration period.

4. Detection indicators and methods:

4.1 Body weight: weigh the body weight with an electronic scale every morning before administration, and record it.

4.2 Open-field experiment: The length, width and height of the open box are 50cm each. The inner wall and bottom of the box are painted black, and the bottom is divided into 25 squares with equal areas. The mice are placed in the middle of the open box to cross The number of squares on the bottom is the score of horizontal movement (stepping in with more than three claws), and the movement of the mouse is observed when both upper limbs leave the box and the measurement time is 5 minutes.

4.3 Sugar water consumption experiment: After the mice were deprived of water each time, the preference degree of 1% sucrose water was measured for each group of animals within 12 hours. Sucrose preference = sucrose water consumption/(sucrose water consumption + distilled water consumption) × 100%.

4.4 Forced swimming test: The forced swimming test was performed on the 21st day of chronic stress. Put the mice into glass tanks with a water temperature of 15°C and a water depth of 15 cm, so that the hind limbs of the mice cannot be supported by the bottom of the glass tank. Each mouse swims for 5 minutes in advance, and observes the swimming behavior for 5 minutes after 24 hours. Record the immobility time and swimming time.

5. Statistical processing: SPSSl6.0 statistical software was used to analyze the data. Metric data with express. Analysis of variance (F test) was used for the significance test between the means of multiple groups, and a significant test (t test) was used between the means of two groups.

6. Test results

6.1 Body weight: the results are shown in Table 1

Table 1: Effects of Lactucoside B on Body Weight of Depressed Mice ( n=10)

group Dose (μg/10g) weight (g) normal control group - 30.27±3.14 depression group - 24.16±2.87 ^** Lettucoside B low dose group 12.5 27.36±0.15 ^△ Lettucoside B medium dose group 25 29.80±1.27 ^△△ Lettucoside B high dose group 50 32.10±1.59 ^△△ Fluoxetine hydrochloride group 35 29.83±0.94 ^△△

Note: Compared with the normal control group ^** P<0.01, compared with the depression group ^△ P<0.05, ^△△ P<0.01

The results in Table 1 show that compared with the normal control group, the weight of the depression group decreased significantly, and the difference was statistically significant (P<0.01); compared with the depression group, the body weight of each dose group of lettucoside B and the fluoxetine group increased significantly , the difference was statistically significant (P<0.05, P<0.01). There was no statistical difference between each dose group and the fluoxetine group.

The results suggest that Lactucoside B can increase the body weight of depressed mice, and this effect is significantly different from that of fluoxetine.

6.2 Field experiment results: as shown in Table 2

Table 2: Effects of Lactucoside B on the results of field experiments in depression mice ( n=10)

group Dose (μg/10g) Number of horizontal movements Upright times normal control group - 28.75±2.10 13.25±1.57 depression group - 16.94±1.32 ^** 8.36±1.45 ^** Lettucoside B low dose group 12.5 19.30±3.52 ^△ 9.26±0.18 Lettucoside B medium dose group 25 26.31±3.40 ^△△ 12.47±1.94 ^△△ Lettucoside B high dose group 50 32.00±4.21 ^△△ 14.58±2.36 ^△△ Fluoxetine hydrochloride group 35 30.78±5.06 ^△△ 14.09±2.11 ^△△

Note: Compared with the normal control group ^** P<0.01, compared with the depression group ^△ P<0.05, ^△△ P<0.01

The results in Table 2 show that compared with the normal control group, the number of horizontal movements and the number of uprights in the depression group were significantly reduced (P<0.01); compared with the depression group, the times of horizontal movements were significantly increased in each dose group of lettucein B, Lactucoside B25μg/10g dose group and 50μg/10g dose group can significantly increase the number of uprights in depressed mice, and the effect of lettucoside B on the number of horizontal movements and the number of uprights in mice is dose-dependent. The effects of the lettucoside B50μg/10g dose group and the fluoxetine group on the field experiment were equivalent.

The results suggest that: Lactucoside B has the effect of increasing the exercise performance of depressed mice.

6.3 Results of sugar water consumption experiment: as shown in Table 3.

Table 3: Effects of Lactucoside B on Sucrose Preference in Depressed Mice ( n=10)

group Dose (μg/10g) Sucrose preference (%) normal control group - 98.25±2.46 depression group - 67.33±5.04 ^** Lettucoside B low dose group 12.5 81.49±7.06 ^△△ Lettucoside B medium dose group 25 87.26±9.52 ^△△ Lettucoside B high dose group 50 95.23±4.37 ^△△ Fluoxetine group 35 92.30±5.11 ^△△

 Note: ^** P<0.01 compared with normal control group, ^△△ P<0.01 compared with depression group

The results in Table 3 show that: compared with the normal control group, the sucrose preference in the depression group was significantly reduced (P<0.01), which proved that the mouse depression model was simulated successfully; The degree of sucrose preference in the glycoside B25μg/10g group and the lettucoside B50μg/10g group increased significantly, P<0.01. The effect was comparable to that of the fluoxetine group.

6.4 Results of forced swimming experiment: as shown in Table 4

Table 4: Effects of Lactucoside B on the results of the forced swimming test in depressed mice ( n=10)

group Dose (μg/10g) Immobility time (s) Swimming time (s) normal control group - 128.30±14.65 169.44±12.39 depression group - 181.37±10.22 ^* 107.43±9.43 ^** Lettucoside B low dose group 12.5 164.31±13.42 ^△ 129.57±2.16 Lettucoside B medium dose group 25 136.31±3.40 ^△ 142.35±10.85 ^△ Lettucoside B high dose group 50 129.07±4.21 ^△△ 157.39±7.30 ^△△ Fluoxetine group 35 130.78±7.04 ^△△ 166.02±23.18 ^△△

Note: Compared with normal control group, ^* P<0.05, ^** P<0.01, compared with depression group, ^△ P<0.05, ^△△ P<0.01

The results in Table 4 show that compared with the normal control group, the swimming immobility time of the mice in the depression group was significantly prolonged (P<0.05), and the swimming time was significantly shortened (P<0.05); It can shorten the swimming immobility time of mice, and the lettucoside B25μg/10g group, the lettucoside B50μg/10g group and the fluoxetine group can prolong the mouse swimming time, and the lettucoside B50μg/10g dose group and the fluoxetine group have no effect on the depressed mice. Forced swim time has an equivalent effect.

The results suggest that Lactucoside B can prolong the swimming time and shorten the swimming time of depressed mice.

Example 2: Preventive effect of Lactucoside B on depression in mice

1. Experimental animals: 50 male Kunming mice, weighing 20-22 g, were purchased from the Experimental Animal Center of Zhengzhou University.

2. Experimental reagents and instruments:

Lactucoside B (prepared by Xinxiang Medical College School of Pharmacy, purity greater than 95%), fluoxetine hydrochloride (Eli Lilly Suzhou Pharmaceutical Co., Ltd.), swimming pool, Open·Field open box.

3. Experimental grouping and experimental methods:

Before the experiment, 50 mice were adaptively fed for 1 week with 12 hours of light and 12 hours of no light, and then randomly selected 10 mice as the normal control group, 20 mice as the model group, and 20 mice as the lettucoside B prophylaxis group. The normal control group was divided into two cages and housed in groups, with 5 animals in each cage. Feed and drinking water were given normally according to the needs, and no stimulation was accepted. The rest of the mice were modeled as follows:

Mice were housed in a single cage and received seven different chronic stress stimuli at the same time. Including: overnight lighting deprivation of sleep environment, fasting (24h), water deprivation (24h), ice water swimming (4°C, 5min), hot water swimming (40°C, 5min), tail pinch (2min), tail suspension (3min) ). One kind of stimulus was randomly arranged every day, and the same kind of stimulus appeared discontinuously. In the 21 days of the experiment, each kind of stimulus was used 3 times on average, so that the mice could not predict the upcoming stimulus.

Model group: During the 21 days of the experiment, the mice in this group were fed in single cages and received various chronic stress stimuli. From the first day of stimulation, 0.1ml/10g of normal saline was injected intraperitoneally at 8:00 a.m. every day.

Lactucoside B preventive medication group: the feeding environment and stimulation methods were the same as those of the depression group, and 25 μg/10 g intraperitoneally injected 0.5 ml of Lactucoside B dissolved in sterilized double distilled water at 8:00 a.m. from the first day of stimulation for 21 consecutive days ;

4. Detection indicators and methods:

4.1 Mortality: At the end of the 21-day experiment, the number of dead animals was counted, and the mortality rate was calculated.

4.2 Body weight: weigh the body weight with an electronic scale every morning before administration, and record it.

4.3 Open-field experiment: the length, width and height of the open box are 50cm each. The inner wall and bottom of the box are painted black. The number of squares on the bottom is the score of horizontal movement (stepping in with more than three claws), and the movement of the mouse is observed when both upper limbs leave the box and the measurement time is 5 minutes.

4.4 Forced swimming test: The forced swimming test was performed on the 22nd day of chronic stress. Put the mice into glass tanks with a water temperature of 15°C and a water depth of 15 cm, so that the hind limbs of the mice cannot be supported by the bottom of the glass tank. Each mouse swims for 5 minutes in advance, and observes the swimming behavior for 5 minutes after 24 hours. Record the immobility time and swimming time.

5. Statistical processing: SPSSl6.0 statistical software was used to analyze the data. Metric data with express. Analysis of variance (F test) was used for the significance test between the means of multiple groups, and a significant test (t test) was used between the means of two groups.

6. Test results

6.1 Mortality: The results are shown in Table 5

Table 5: Effects of Lactucoside B on the Mortality of Depressed Mice

group Dose (μg/10g) number of deaths Mortality (%) normal control group - 0 0 model group - 8 40 ^** Lactucoside B prophylaxis group 25 0 0 ^△△

Note: ^** P<0.01 compared with normal control group, ^ΔΔ P<0.01 compared with model group

The results in Table 5 show that compared with the normal control group, the model group had a higher mortality rate of 40% during the modeling process, and the mouse mortality rate was significantly reduced after the prophylactic injection of lettuce B to the mice (P<0.01) , compared with the normal control group, there was no significant difference.

The results suggest that: Lactucoside B can reduce the mortality of depression mice.

6.2 Body weight: the results are shown in Table 6

Table 6: The effect of prophylactic administration of Lactucoside B on body weight of mice with depression

group Dose (μg/10g) weight (g) normal control group - 29.15±2.63 model group - 22.40±3.81 ^* Lactucoside B prophylaxis group 25 30.72± ^1.89Δ

Note: ^* P<0.05 compared with the normal control group, ^Δ P<0.05 compared with the model group

The results in Table 6 show that compared with the normal control group, the weight of the model group decreased significantly, and the difference was statistically significant (P<0.05); compared with the model group, the body weight of the lettucoside B preventive medication group increased significantly, and the difference was statistically significant Significance (P<0.05)

The results suggest that the prophylactic administration of Lactucoside B can increase the body weight of depressed mice.

6.3 Field experiment results: as shown in Table 7

Table 7: Effects of preventive medication of Lactucoside B on the results of field experiments in depressive mice

group Dose (μg/10g) Number of horizontal movements Upright times normal control group - 26.35±1.04 12.60±1.33 model group - 13.76±2.09 ^** 7.19±0.85 ^** Lactucoside B prophylaxis group 25 27.82±2.46 ^△△ 13.55±1.94 ^△△

 Note: ^** P<0.01 compared with normal control group, ^△△ P<0.01 compared with model group

The results in Table 7 show that: compared with the normal control group, the number of horizontal movements and the number of uprights in the model group were significantly reduced, P<0.01; Number of upright mice.

The results suggest that the prophylactic drug of lettucein B can increase the exercise performance of depressed mice.

6.4 Results of forced swimming experiment: as shown in Table 8

Table 8 The effect of prophylactic medication of Lactucoside B on the results of forced swimming test in depressed mice

group Dose (μg/10g) Immobility time (s) Swimming time (s) normal control group - 119.87±6.43 171.20±9.35 model group - 186.01±4.70 ^* 96.22±7.05 ^** Lactucoside B prophylaxis group 25 126.14±10.68 ^△ 164.99±11.37 ^△△

Note: ^* P<0.05 compared with the normal control group, ^△△ P<0.01 compared with the model group

The results in Table 8 show that compared with the normal control group, the swimming immobility time of the mice in the model group was prolonged and the swimming time was shortened, P<0.05; the lettucoside B preventive drug group could shorten the swimming immobility time of the mice and prolong the swimming time of the mice. time.

The results suggest that preventive medication of Lactucoside B can prolong the swimming time and shorten the immobility time of depressed mice.

Experimental summary of experimental examples 1 and 2:

According to the experimental results of Example 1, it is suggested that Lactucoside B can increase the body weight of depressed mice, increase its exercise performance, prolong its swimming time, and improve its preference for sucrose water. The above effects all show a dose-dependent relationship, and its The effect is equivalent to that of fluoxetine, an antidepressant commonly used in clinical practice, suggesting that lettucoside B has a therapeutic effect on depression;

According to the experimental results of Example 2, it is suggested that the preventive use of Lactucoside B can significantly reduce the mortality rate of depression mice, increase its body weight and exercise performance, and prolong the swimming time of animals, suggesting that Lactucoside B has a better preventive effect on depression. .

The above results and conclusions were drawn on the basis of using various mild stimuli to induce depression in mice. In view of this, Lactucoside B has obvious preventive and therapeutic effects on mental depression induced by various reasons. Depression induced by other reasons cannot be judged temporarily due to limitations of technical means, but according to the current experimental results, the effect of Lactucoside B on animal activities suggests that it also has a certain preventive effect on depression induced by other reasons .

Although, the present invention has been described in detail with general description, specific implementation and test above, but on the basis of the present invention, some modifications or improvements can be made to it, which will be obvious to those skilled in the art . Therefore, the modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the protection scope of the present invention.

Claims (6)

1. Application of Lactucoside B in the preparation of medicaments for preventing and treating depression. 2. The application according to claim 1, characterized in that, the medicine is a preparation containing Lactucoside B. 3. The application according to claim 2, characterized in that the preparation is made from raptoside B alone or from raptoside B and a pharmaceutically acceptable carrier. 4. The application according to claim 3, wherein the preparation is tablet, capsule, pill, oral liquid, granule or injection. 5. The application according to claim 3, wherein the pharmaceutically acceptable carrier refers to a conventional drug carrier in the field of pharmacy, selected from fillers, binders, disintegrants, lubricants, solubilizers , suspending agent, wetting agent, pigment, essence, solvent, surfactant or flavoring agent or one or more of them. 6. The application according to any one of claims 1-5, characterized in that the proposed clinical dosage of Lactucoside B is: 100-400 mg/d.