Application of Eleutheroside B in the preparation of drugs for preventing and / or treating methamphetamine addiction withdrawal

Abstract

The present invention belongs to the field of biomedicine, and discloses the application of eleutheroside B in the preparation of drugs for preventing and / or treating methamphetamine addiction withdrawal. The inventor evaluated the role of eleutheroside B in preventing and treating methamphetamine addiction by investigating the effects of eleutheroside B on methamphetamine-induced conditioned place preference and behavioral sensitization. It was experimentally confirmed that eleutheroside B could reverse methamphetamine-induced conditioned place preference and reduce methamphetamine-induced hyperlocomotor activity, and had an inhibitory effect on the expression of methamphetamine-induced behavioral sensitization in mice, indicating that eleutheroside B could effectively weaken the dependence and craving for methamphetamine, weaken the induced behavioral sensitization effect, and could be used for preventing and treating methamphetamine addiction. The present invention develops a new drug for drug rehabilitation with eleutheroside B as the active ingredient, provides an effective and safe option for the prevention and treatment of methamphetamine addiction, and has good clinical application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine and relates to a new use of eleutheroside B, and specifically, to the use of eleutheroside B in the preparation of a drug for preventing and / or treating methamphetamine addiction. Background Art

[0002] Methamphetamine (Meth) is a type of amphetamine stimulant, commonly known as "ice", and is a solid white crystal with a molecular formula of C 10 H 15 N.

[0003] Methamphetamine will force the corresponding neurons to release excessive amounts of dopamine, causing abnormal stimulation to the nerves and making the user addicted. At the same time, methamphetamine will prevent the repeated use of dopamine. Methamphetamine is a positive lipophilic molecule that can quickly enter the mitochondria of brain neurons, breaking the balance of cations inside the mitochondria, leading to DNA breakage, chromatin condensation, nuclear fragmentation, and ladder-like fragmentation of DNA, inducing neuronal death and causing irreversible damage.

[0004] Methamphetamine addiction has become a global problem that needs to be solved urgently. Methamphetamine addiction is a chronic and recurrent brain disease, which is mainly characterized by the strong desire for drugs, compulsive drug-seeking psychology and protracted withdrawal symptoms after drug discontinuation. Long-term use of methamphetamine inhibits the metabolism and reuptake of dopamine, promotes the synthesis of dopamine, compensatory increases in the dopamine level in reward-related brain areas, enhances the function of the dopamine system, produces reward effects and mental dependence, and behaviorally causes behavioral sensitization, physical and mental dependence and relapse behavior in the body. At present, the treatment of methamphetamine uses methadone treatment, which "replaces large poison with small poison", but methadone itself is also a drug. Studies have reported that patients who use methadone to treat methamphetamine addiction need to take methadone for life, which cannot fundamentally solve the harm of methamphetamine to the human body. Therefore, it is of great practical significance to seek drugs that have definite efficacy, no toxic reactions, and no addiction to prevent and treat methamphetamine addiction.

[0005] Eleutheroside B (EleB) is a chemical component extracted from the root or stem of Acanthopanax senticosus of the Araliaceae family. Its structure is as follows:

[0006]

[0007] The traditional Chinese medicine Acanthopanax senticosus was first recorded in "Shennong Ben Cao Jing" and has the effects of replenishing qi and strengthening the spleen, tonifying the kidney and calming the mind. The record of Acanthopanax senticosus in "Ming Yi Bie Lu" is that "it can strengthen the muscles and bones, strengthen the will, and long-term use can make the body light and endure fatigue". Modern research shows that the chemical components in Acanthopanax senticosus are mainly Acanthopanax saponins, and Acanthopanax saponin B is the main active monomer of Acanthopanax saponins. Acanthopanax saponin B has antidepressant, anti-Alzheimer's disease, anti-fatigue, anti-inflammatory, antioxidant and neuroprotective functions. Recent research reports that Acanthopanax saponin B can delay neuronal apoptosis by promoting neuronal function recovery. Acanthopanax saponin B can also reduce the sleep latency and prolong the sleep time of mice, and its molecular mechanism of action may be related to the NOS / NO pathway. In the aspect of anti-Alzheimer's disease, some studies have shown that Acanthopanax saponin B has a protective function on damaged neurons in Alzheimer's disease and may play a role by alleviating amyloid-β-induced cerebral cortical neuron atrophy. In the aspect of antidepressant, some studies believe that it may treat depression through multiple aspects such as regulating BDNF in related brain regions, repairing damaged neurons, scavenging free radicals, antioxidant and increasing the monoamine neurotransmitter 5-HT. However, whether Acanthopanax saponin B can play a therapeutic role in methamphetamine addiction has not been reported at home and abroad. Summary of the Invention

[0008] The inventors investigated the effects of Acanthopanax saponin B on methamphetamine-induced conditioned place preference and behavioral sensitization to evaluate the role of Acanthopanax saponin B in the prevention and treatment of methamphetamine addiction. It was experimentally confirmed that Acanthopanax saponin B can reverse methamphetamine-induced conditioned place preference and reduce methamphetamine-induced hyperactivity, and has an inhibitory effect on the expression of methamphetamine-induced behavioral sensitization in mice, indicating that Acanthopanax saponin B can effectively weaken the dependence and craving for methamphetamine, weaken the induced behavioral sensitization effect, and can be used for the prevention and treatment of methamphetamine addiction.

[0009] The purpose of the present invention is to provide the application of Acanthopanax saponin B in the preparation of drugs for preventing and / or treating methamphetamine addiction withdrawal.

[0010] In the present invention, addiction refers to behaviors related to addiction, mainly including compulsive drug use and drug seeking, relapse, and increased behavioral activity, which are manifested as conditioned place preference (CPP) model and behavioral sensitization model in animal models.

[0011] The methamphetamine addiction withdrawal drug described above is a pharmaceutically acceptable dosage form made of Acanthopanax saponin B as the sole active ingredient or the main active ingredient, together with pharmaceutically acceptable excipients or auxiliary ingredients.

[0012] Furthermore, the methamphetamine addiction withdrawal drug described above may or may not include other active ingredients.

[0013] As a specific embodiment, the methamphetamine addiction withdrawal drug uses eleutheroside B as the sole active ingredient.

[0014] As a specific embodiment, the methamphetamine addiction withdrawal drug uses eleutheroside B and other active ingredients with therapeutic or adjuvant therapeutic effects on methamphetamine addiction withdrawal as active ingredients.

[0015] The other active ingredients with therapeutic or adjuvant therapeutic effects on methamphetamine addiction withdrawal can be selected from methadone.

[0016] In the present invention, the effective dose of eleutheroside B is 50 mg / kg to 200 mg / kg, and the optimal dose is 200 mg / kg.

[0017] The pharmaceutically acceptable excipients include, but are not limited to, conventional pharmaceutically used fillers, diluents, binders, excipients, penetration enhancers, absorption enhancers, surfactants, pH regulators, antioxidants, stabilizers, chelating agents, pore-forming agents, film-forming agents, solvents, cosolvents, flavoring agents, sweetening agents, pigments, etc.

[0018] The auxiliary components include, but are not limited to, sodium hyaluronate injection, oxaliplatin for injection, etc.

[0019] The drugs include, but are not limited to, dosage forms for gastrointestinal administration and non-gastrointestinal administration.

[0020] The dosage forms for gastrointestinal administration can be tablets, capsules, soft capsules, liquid capsules, granules, dripping pills, syrups, dry suspensions, oral solutions, oral suspensions, orally disintegrating tablets, orally soluble films, sustained-release tablets, controlled-release tablets, sustained-release capsules, controlled-release capsules, enteric-coated tablets, enteric-coated capsules; the dosage forms for non-gastrointestinal administration can be aqueous injections, lyophilized powder injections, sterile powder injections, infusions, eye drops, nasal drops, nasal sprays, pulmonary inhalants, transdermal patches, liniments, ointments, creams, suppositories, subcutaneous implants, long-acting injections, microneedle preparations. For the above dosage forms, the present invention does not make any limitations here. Any dosage form recognized in medicine is within the protection scope of the present invention.

[0021] The application is the application of eleutheroside B in the preparation of a drug for treating and / or preventing methamphetamine addiction withdrawal, which plays a role in at least one of weakening the dependence on and craving for methamphetamine, reducing dopamine in the NAc brain region, and inhibiting drug-seeking behavior and behavioral sensitization.

[0022] The beneficial effects of the present invention:

[0023] 1) In view of the current situation that there is no FDA-approved drug for the treatment of methamphetamine addiction, the present invention provides a new use of eleutheroside B in drugs for the prevention and / or treatment of methamphetamine addiction, providing a direction for the development of new drug for drug rehabilitation with eleutheroside B as the active ingredient, promoting the development of eleutheroside B in the prevention and / or treatment of methamphetamine addiction withdrawal, and providing an effective and safe option for the prevention and treatment of methamphetamine addiction, with good clinical application prospects.

[0024] 2) By observing the behavioral responses of mice through the conditioned place preference (CPP) model and the behavioral sensitization model, the present invention finds that eleutheroside B has an inhibitory effect on the relapse and sensitization behaviors of mice, suggesting that eleutheroside B has a rapid, comprehensive and significant effect on reversing methamphetamine addiction. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is the experimental flow chart of continuously intragastrically administering eleutheroside B to mice after establishing the methamphetamine conditioned place preference (CPP) model and withdrawal; wherein, SA represents normal saline, MA represents methamphetamine, EleB represents eleutheroside B, Withdrawperiod represents the withdrawal state of mice after establishing the methamphetamine conditioned place preference model, Test1 represents baseline test 1, Test2 represents modeling score test 2, and Test3 represents the scoring test 3 for the therapeutic effect of eleutheroside B.

[0026] Figure 2 It is the influence of different doses of eleutheroside B on the formation of methamphetamine-induced conditioned place preference; wherein, A is the conditioned place preference score of different doses of eleutheroside B in the treatment of methamphetamine addiction, B is the representative behavioral trajectory of the high-dose group of eleutheroside B in the treatment of methamphetamine addiction; control represents the control group, model represents the methamphetamine model group, EleB-L represents the low-dose group of eleutheroside B (50 mg / kg), EleB-M represents the medium-dose group of eleutheroside B (100 mg / kg), EleB-H represents the high-dose group of eleutheroside B (200 mg / kg), and two-way ANOVA is used for statistical analysis, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns indicates no significant statistical difference.

[0027] Figure 3It is the effect of different doses of eleutheroside B on dopamine in the nucleus accumbens (NAC) of the mouse brain regions related to addiction. Among them, Control represents the control group, Model represents the methamphetamine model group, EleB-L represents the low-dose eleutheroside B administration group (50 mg / kg), EleB-M represents the medium-dose eleutheroside B administration group (100 mg / kg), and EleB-H represents the high-dose eleutheroside B administration group (200 mg / kg). One-way ANOVA was used for statistical analysis. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, and ns indicates no significant statistical difference.

[0028] Figure 4 It is the experimental flow chart of the effect of eleutheroside B on the occurrence, development, and expression of methamphetamine-induced behavioral sensitization in mice. Among them, Baseline is the habituation training for three consecutive days, MA is methamphetamine, EleB is eleutheroside B, Formative period is the methamphetamine behavioral sensitization formation plus treatment period, and Withdraw period is the withdrawal period.

[0029] Figure 5 It is the baseline determination on the last day of the three-day habituation training of mouse behavioral sensitization. Among them, A is the total movement distance of behavioral activities in 30 min, B is the movement distance in each 5-min time period during the baseline period, and C is the behavioral trajectory of the mouse within 30 min. Control represents the control group, Model represents the model group, and EleB represents the eleutheroside B group.

[0030] Figure 6 It is the effect of eleutheroside B on the behavioral activities of mice after a single administration of methamphetamine on the first day. Among them, A is the total movement distance of behavioral activities in 1 h, B is the movement distance in each 5-min time period, and C is the behavioral trajectory of the mouse within 1 h (the first 30 min is the spontaneous activity of the mouse, and the last 30 min is the behavioral activity of the mouse after methamphetamine administration). Control represents the normal control group, Model represents the model group, and EleB represents the eleutheroside B group. One-way ANOVA was used for statistical analysis. * represents comparison between the model group and the control group, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; # represents comparison between the eleutheroside B group and the model group, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001.

[0031] Figure 7The role of eleutheroside B in the occurrence and development of methamphetamine addiction in mice over 8 consecutive days; among them, A is the total movement distance of the behavioral activities of mice for 30 minutes per day, and B is the effect of eleutheroside B on methamphetamine expression on the 21st day; Control represents the control group, Model represents the model group, and EleB represents the eleutheroside B group. One-way ANOVA was used for statistical analysis. * represents a comparison between the model group and the control group, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; # represents a comparison between the eleutheroside B group and the model group; #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001. Specific implementation manner

[0032] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the following further explains the present invention in detail in combination with the implementation manners. Without departing from the concept of the present invention, all similar substitutions and modifications are obvious to those skilled in the art, and these can be regarded as included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and those skilled in the art can obviously make changes or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of the present invention to implement and apply the technology of the present invention.

[0033] Example 1

[0034] 1. Materials and drugs

[0035] 1.1 Experimental animals: C57BL / 6J mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., SPF grade, male, with a body weight of 16 - 19 g, and provided with a Jiangsu experimental animal facility use certificate by the Experimental Animal Center of China Pharmaceutical University. The temperature was (23 ± 1) °C, and the humidity was (50 ± 10)%. All experimental operations related to animals complied with the requirements of the Medical Ethics Committee of China Pharmaceutical University.

[0036] 1.2 Drugs:

[0037] 1) Methamphetamine standard was provided by the Anti - drug Intelligence Technology Center of the Ministry of Public Security of China, Beijing. The standard was dissolved in physiological saline and administered by intraperitoneal injection.

[0038] 2) Eleutheroside B standard was purchased from Nanjing Dilger Co., Ltd., dissolved in 0.5% CMC - Na, and administered by gavage.

[0039] 1.3 Experimental instruments:

[0040] Conditioned Place Preference Box (Jiangsu Sians Biotechnology Co., Ltd.), as a behavioral training instrument and detection instrument. The Conditioned Place Preference Box is divided into two chambers: a black chamber (drug-paired chamber, non-natural preference area) and a white chamber (non-drug-paired chamber, preference area). There is a small door between the two chambers through which mice can freely shuttle, and it can be closed or connected.

[0041] Animal Spontaneous Activity Video Analysis System, used to record the movement behavior of mice. This system consists of an open field (length × width × height: 45 cm × 45 cm × 45 cm) and ananymaze software.

[0042] 2. Experimental Methods

[0043] Experiment 1: The conditioned place preference (CPP) behavioral method and the neurotransmitter dopamine (DA) detection method were used to explore the effect of eleutheroside B on the drug-seeking behavior of methamphetamine-addicted mice.

[0044] Experimental mechanism: CPP is currently a classic model for evaluating drug psychological dependence and is also the most widely used and effective tool for anti-drug-seeking behavior. In this experiment, the experimental animals were placed in the non-natural preference area of the conditioned place preference box, and a psychologically dependent drug (methamphetamine was given in this experiment) was administered. Then, the activities of the experimental animals in the non-preference area and the preference area of the conditioned place preference box were observed. Each time the experimental animals were in the drug administration area, they would develop a positional preference for the black and white box areas under the action of the drug reward effect, and the degree was related to drug dependence. Using the CPP model to simulate the clinical withdrawal state as a mouse drug-seeking behavior model can reflect the behavior of methamphetamine-seeking (re-addiction) in mice during the methamphetamine withdrawal period when they are re-exposed to the environment related to previous methamphetamine use or methamphetamine itself.

[0045] The excitatory behavior produced by methamphetamine, as well as the craving and drug-seeking behavior for the drug, are all related to the dopamine neurotransmitter in the brain. Methamphetamine can cause the axon terminals of NAc neurons to release dopamine and activate neurons in related brain regions such as the hippocampus and striatum, which will cause changes in the body's cognitive ability and psychoactivity. Methamphetamine can also act as a false neurotransmitter, bind to the dopamine transporter at the dopamine neuron terminals, and promote the depletion release of DA in the vesicles, resulting in a sharp increase in the DA content in the synaptic cleft of the neuron axons. When methamphetamine-addicted mice are in a period of withdrawal state and are placed again in an environment related to addiction reward, it will activate the reward system of the mice. The biological basis of this "expectancy effect" is dopamine, and the stronger the reward cue received, the more vigorous the secretion of dopamine.

[0046] Experimental method: Forty male C57BL / 6J mice (4 weeks old, weighing 16 - 19 g) were housed in a SPF-class animal room and adaptively fed for 1 week. They were randomly divided into a control group (8 mice) and a modeling group (32 mice). The experimental flow chart is shown in Figure 1 , and on the subsequent day 1, baseline measurement (Test1) was performed: The mice were placed in the middle chamber, the small door was opened, and the mice were allowed to freely shuttle back and forth between the two side chambers for 15 min. The time the mice stayed in the black and white chambers was statistically analyzed by the anymaze software. After the baseline measurement was completed, the natural preference chamber for the mice was selected as the white chamber, and then methamphetamine conditioned place preference modeling training was carried out. On day 2, the mice in the control group and the modeling group were intraperitoneally injected with normal saline and placed in the white chamber for training for 30 min. On day 3, the mice in the control group were intraperitoneally injected with normal saline, and the mice in the modeling group were injected with methamphetamine (2 mg / kg / d) and placed in the black chamber for training for 30 min. The operations of the previous two days were repeated in the following 6 days, and methamphetamine addiction modeling was performed on the mice for 8 consecutive days by administering the drug every other day. Then, a modeling score test (Test2) was carried out. After the modeling was completed, the mice in the modeling group were randomly divided into a methamphetamine model group (Model), a low-dose eleutheroside B group (ELeB-L), a medium-dose eleutheroside B group (ELeB-M), and a high-dose eleutheroside B group (ELeB-H), with 8 mice in each group. Methamphetamine withdrawal was performed on the mice. During the withdrawal process, the mice in the low-, medium-, and high-dose eleutheroside B groups were intragastrically administered drugs every day, and the daily drug administration doses were 50 mg / kg, 100 mg / kg, and 200 mg / kg, respectively. The mice in the control group and the methamphetamine model group were given 0.5% CMC-Na with the same volume as the drug administration group. Then, a recovery drug-seeking test (Test3) was carried out. After the experiment was completed, the nucleus accumbens (NAc) brain tissues of the mice were collected for the detection of the neurotransmitter dopamine (DA).

[0047] 1) Score the conditioned place preference of the mice:

[0048] Test1 was carried out on the first day after the mice were adapted. The mice were placed in the conditioned place preference chamber, which was in a connected state, and allowed to freely move for 900 s. The activity time of the mice in the drug-paired chamber was analyzed and recorded as the baseline value. Starting from the second day, for the subsequent 8 days, methamphetamine addiction training was carried out by administering the drug every other day. After administering the drugs according to the above grouping requirements, the mice were restricted in the drug-paired chamber (closed state) for training for 30 min / d. After the training was completed, on day 10, the mice were placed in the experimental instrument and allowed to freely move (connected state) for 900 s, and the activity time of the mice in the two side chambers was recorded. Then, during the methamphetamine withdrawal state of the mice, eleutheroside B was continuously intragastrically administered to them. On day 23, the third behavioral test (Test3) was carried out. Then, the activity time of the mice in the drug-paired chamber was subtracted from the activity time of the mice in the non-drug-paired chamber to obtain the difference, and this difference was the conditioned place preference score (CPP Score).

[0049] 2) Statistical analysis was performed as follows:

[0050] Data were uniformly expressed as "mean ± standard deviation", and two-way ANOVA was performed using Graph prism 9.0 software to compare the differences between groups ( Figure 2 ), and one-way ANOVA was used to compare the differences between groups ( Figure 3 ).

[0051] Experimental results: As Figure 2 shown in A, except for the control group, there were significant differences in the conditioned place preference scores of mice on the 10th day compared with those on the 1st day, indicating successful modeling in the model group; there was no difference in the conditioned place preference scores of mice in the control group on the 23rd day compared with those on the 10th day, and the conditioned place preference scores of mice in the methamphetamine model group increased slightly compared with those on the 10th day. However, the conditioned place preference scores of mice in each dose group of eleutheroside B on the 23rd day decreased significantly compared with those on the 10th day and reversed the positive value of the conditioned place preference scores on the 10th day, indicating that eleutheroside B has the effect of reversing methamphetamine addiction. And it can be seen that the treatment effect of the eleutheroside B group is: EleB-L < EleB-M < EleB-H, indicating that eleutheroside B reverses methamphetamine addiction in a dose-dependent manner, and eleutheroside B inhibits methamphetamine-seeking behavior behaviorally. Figure 2 B is a representative diagram of the behavioral trajectory of the high-dose group of eleutheroside B (EleB-H) in mice. On the 1st day, the mice showed obvious preference for the right side (white box) during the baseline period. On the 10th day, after modeling, the modeled mice showed obvious preference for the left side (black box). On the 23rd day, after treatment with eleutheroside B, the mice in the drug administration group re-preferred the right side (white box), visually showing the situation of eleutheroside B reversing methamphetamine addiction.

[0052] After the methamphetamine model group completed modeling and withdrawal, the time of the model group mice in the drug-associated box was higher than that in the non-drug-associated box. And as Figure 3 shown, the content of the neurotransmitter DA in the NAc brain region of the methamphetamine model group mice was significantly higher than that of the control group. Eleutheroside B decreased the neurotransmitter DA in the NAc brain region of mice in a dose-dependent manner, indicating that eleutheroside B may play a role in reducing methamphetamine-seeking behavior in methamphetamine-addicted mice by reducing dopamine in the NAc brain region.

[0053] Experiment 2: The behavioral sensitization model was used to explore the prevention of eleutheroside B on methamphetamine-addicted mice and the therapeutic effect of eleutheroside B in the occurrence and development of methamphetamine addiction in mice.

[0054] The behavioral sensitization experiment is one of the commonly used models for studying addictive drugs at present. It is significantly different from the pharmacological characteristics of other drugs. Prolonged intermittent use of psychoactive drugs will cause reverse tolerance to the drugs, which is also known as drug sensitization. The most easily observed change in drug sensitization is the behavioral change, such as the increase in spontaneous activity, which is also known as behavioral sensitization.

[0055] Evaluate behavioral sensitization:

[0056] The behavioral characteristics of the behavioral sensitization effect produced by mice are manifested as a significant increase in spontaneous activity, which is mainly reflected by the activity distance generated. The movement behavior of mice is recorded by an animal spontaneous activity video analysis system.

[0057] Adopt statistical analysis:

[0058] The data representation is uniformly specified as "mean ± standard deviation", and one-way ANOVA is used with Graph prism 9.0 software to compare the differences among groups.

[0059] Experimental method: As Figure 4 , take 21 C57BL / 6J mice and randomly divide them into 3 groups: control group, model group, and eleutheroside B group (dose 200 mg / kg); 1), the mice in the three groups are continuously trained for habituation for 3 days. Every day, the mice are placed in an open field to adapt for 30 min. The last day is used as the baseline level, measured for 30 min, and data is recorded every 5 min. Record the spontaneous activity level of each group of mice measured during the baseline period. 2), the mice in the eleutheroside B group are given 200 mg / kg eleutheroside B by gavage, and the mice in the control group and the model group are given the same volume of CMC-Na by gavage. 1.5 h later, the mice are placed in the open field and the spontaneous activity of the mice within 30 min is recorded. Then, the mice in the eleutheroside B group and the model group are taken out and methamphetamine is intraperitoneally injected at a dose of 2 mg / kg. The control group of mice is intraperitoneally injected with the same volume of normal saline, and the mice are immediately placed in the open field to record the motor activity 30 min after administration. 3), repeat the process in 2) continuously for 8 days (1d - 8d), and stop the drug for 12 days during the withdrawal period (9d - 20d, without any treatment). On the 21st day, methamphetamine is intraperitoneally injected into the mice in the methamphetamine group and the eleutheroside B group (dose 1 mg / kg), and the control group is injected with the same volume of normal saline. The mice are immediately placed in the open field to record the motor activity of the mice within 30 min after administration, and the movement distance is recorded every 5 min as a time period.

[0060] Experimental results:

[0061] 1) The spontaneous activity levels of each group of mice measured during the baseline period

[0062] During the Baseline stage ( Figure 4There were no significant differences in the spontaneous activities of each group of mice, as shown in Figure 5 . Figure 5 A shows that there were no differences in the total movement distance of each group of mice in 30 min (P>0.05). Figure 5 B shows that there were no significant differences in the movement distance of mice every 5 min within 30 min of the total movement time (P>0.05). Figure 5 C shows that there were no obvious differences in the movement behavior trajectories of each group of mice.

[0063] 2) Effects of single-dose administration of eleutheroside B on the enhanced hyperactivity effect in mice caused by acute methamphetamine treatment

[0064] Figure 6 A shows the total movement distance of the behavioral activities of each group of mice within 1 h. Compared with the control group, the movement distance of the mice in the model group increased significantly (P<0.01); compared with the model group, the movement distance of the mice in the eleutheroside B group decreased significantly, indicating that eleutheroside B can significantly reduce the increase in locomotor activity caused by methamphetamine. Figure 6 B shows the movement distance every 5 min within 60 min. It can be seen that before methamphetamine administration (0 min), there were no statistical differences in the movement distance of each group of mice (P>0.05), indicating that eleutheroside B itself does not affect the locomotor activity of mice. Figure 6 C shows the behavioral activity trajectories of mice for 1 h. After intraperitoneal injection of methamphetamine (2 mg / kg), compared with the control group, the movement activity distance of the mice in the model group increased significantly. In the eleutheroside B (200 mg / kg) group, 15 min after methamphetamine administration, compared with the methamphetamine group, it can significantly reduce the enhanced locomotor activity effect in mice caused by methamphetamine treatment.

[0065] 3) Role of continuous administration of eleutheroside B for 8 days in the occurrence and development of methamphetamine addiction in mice and effects of eleutheroside B on the expression of methamphetamine-induced behavioral sensitization in mice

[0066] As Figure 7 shown in A, the daily spontaneous activity counts of the mice in the model group were significantly higher than those in the control group (P<0.001), indicating that the mouse behavioral sensitization model had been established. Compared with the model group, the daily spontaneous activity counts of the mice in the eleutheroside B group were significantly reduced every day (except on the 3rd day), indicating that eleutheroside B can play a certain therapeutic role in the occurrence and development of methamphetamine addiction in mice. Figure 7Effect of eleutheroside B on methamphetamine addiction expression on the 21st day. Compared with the control group, the locomotor activity distance of the model group mice increased significantly (P<0.001), indicating that methamphetamine administered at a dose of 1 mg / kg could induce an increase in hyperlocomotor activity in mice. Compared with the methamphetamine model group, eleutheroside B group mice could significantly reduce the effect of increased locomotor activity in mice caused by methamphetamine treatment (P<0.05), indicating that eleutheroside B could inhibit the expression of methamphetamine behavioral sensitization in mice.

[0067] The above experiments showed that eleutheroside B could dose-dependently reduce the methamphetamine CPP score, and correspondingly, eleutheroside B dose-dependently reduced the DA concentration in the nucleus accumbens brain region of mice, indicating that eleutheroside B might reduce the compulsive drug-seeking behavior of mice by reducing the DA concentration in the nucleus accumbens brain region, and 200 mg / kg of eleutheroside B had the best effect. In subsequent behavioral sensitization experiments, 200 mg / kg was selected as the administration dose of eleutheroside B for further exploration. Through the behavioral sensitization experiment, it was found that there were no significant differences in the behavioral activities of each group of mice during the baseline period, and there were no significant differences in the behavioral activities of mice within 30 minutes after administering eleutheroside B for 1 h, indicating that eleutheroside B itself did not produce a sensitization effect, excluding the inhibitory effect of eleutheroside B on behavioral sensitization by inhibiting the activity of mice. In the establishment of a methamphetamine mouse behavioral sensitization model, methamphetamine played a certain role in the occurrence and development of the disease in mice, and it was found on the 21st day that eleutheroside B had an inhibitory effect on the expression of methamphetamine-induced behavioral sensitization in mice. Therefore, eleutheroside B could effectively weaken the dependence, craving, and compulsive drug-seeking behavior on methamphetamine, weaken the induced behavioral sensitization effect, and could treat methamphetamine addiction, with good clinical application prospects.

[0068] The application of eleutheroside B in the prevention and / or treatment of methamphetamine addiction is the first discovery of the inventor. Whether eleutheroside B is used alone as an active ingredient or in combination, the application of eleutheroside B in the prevention and / or treatment of methamphetamine addiction is within the protection scope of the present invention.

[0069] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific embodiments described. Obviously, many modifications and variations can be made according to the content of this specification. These embodiments are selected and specifically described in this specification to better explain the principle and practical application of the present invention, so that those skilled in the art can well understand and utilize the present invention. The present invention is only limited by the claims and their full scope and equivalents.

Claims

1. Use of eleutheroside B in the preparation of a drug for preventing and / or treating methamphetamine addiction withdrawal.

2. The application according to claim 1, wherein: The drug is a pharmaceutically acceptable dosage form prepared from eleutheroside B as the sole active ingredient or the main active ingredient and pharmaceutically acceptable excipients.

3. The application according to claim 2, characterized in that: The drug includes or does not include other active ingredients.

4. The application according to claim 3, wherein: The drug uses eleutheroside B as the sole active ingredient.

5. The application according to claim 3, characterized in that: The drug uses eleutheroside B and other active ingredients with a therapeutic or adjuvant therapeutic effect on methamphetamine addiction withdrawal as active ingredients.

6. The application according to claim 1, characterized in that: The effective dose of eleutheroside B is 50 mg / kg to 200 mg / kg.

7. The application according to claim 1 or 6, characterized in that: The effective dose of eleutheroside B is 200 mg / kg.

8. The application according to claim 2, wherein: The pharmaceutically acceptable excipients include one or more of fillers, diluents, binders, excipients, penetration enhancers, absorption enhancers, surfactants, pH regulators, antioxidants, stabilizers, chelating agents, pore-forming agents, film-forming agents, solvents, cosolvents, flavoring agents, sweetening agents, and pigments.

9. The application according to claim 1, wherein: The drug is a gastrointestinal dosage form and a non-gastrointestinal dosage form; The gastrointestinal dosage form is one or more of tablets, capsules, granules, dripping pills, syrups, dry suspensions, oral solutions, oral suspensions, and oral rapid-dissolving films; The non-gastrointestinal dosage form is one or more of aqueous injections, freeze-dried powder injections, infusion solutions, long-acting injections, eye drops, nasal drops, nasal sprays, pulmonary inhalants, transdermal patches, liniments, ointments, creams, suppositories, subcutaneous implants, and microneedle preparations.

10. The application according to claim 9, characterized in that: The tablet is one of orally disintegrating tablets, sustained-release tablets, controlled-release tablets, and enteric-coated tablets.

11. The application according to claim 9, characterized in that: The capsule is one of soft capsules, liquid capsules, sustained-release capsules, controlled-release capsules, and enteric-coated capsules.

12. The application according to claim 1, wherein: The application is the use of eleutheroside B in the preparation of a drug for treating and / or preventing methamphetamine addiction withdrawal by at least one of weakening the dependence on and craving for methamphetamine, reducing dopamine in the NAc brain region, inhibiting drug-seeking behavior, and behavioral sensitization.

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