Application of parthenolide in preparation of medicine for preventing and / or treating nicotine addiction

Abstract

The invention discloses application of parthenolide in preparation of a medicine for preventing and / or treating nicotine addiction, and belongs to the technical field of medicines. Research finds that different doses of parthenolide can significantly prevent formation of nicotine-induced mouse CPP and formation of nicotine forced foraging; different doses of parthenolide can also significantly improve the withdrawal symptom of nicotine without affecting the spontaneous movement ability of mice, and meanwhile, parthenolide can also prevent nicotine from reburning. Therefore, parthenolide can be used for preparing the medicine for preventing and / or treating nicotine addiction.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a use of parthenolide in preparation of a medicine for preventing and / or treating nicotine addiction. BACKGROUND

[0002] Parthenolide (CAS No.: 20554-84-1) has a molecular formula of C 15 H 20 O3 and a structural formula as shown in the following: .

[0003] Parthenolide belongs to a sesquiterpene lactone compound and is extracted from feverfew. A large number of scientific studies show that parthenolide and its analogues have various pharmacological effects. Parthenolide is traditionally mainly used for treating migraine, fever and rheumatoid arthritis, and in recent years, it has been found to have important effects in anti-tumor, anti-inflammatory, anti-virus and neuroprotection.

[0004] At present, there is no report on the use of parthenolide for treating nicotine addiction. SUMMARY

[0005] The present application aims to provide a use of parthenolide in preparation of a medicine for preventing and / or treating nicotine addiction. The present application researches and finds that different doses of parthenolide can prevent nicotine addiction in mice, and parthenolide significantly improves the anxiety and depression-like behaviors of mice after nicotine withdrawal, and does not affect the spontaneous activity of the mice, and has a potential therapeutic effect on nicotine addiction. Therefore, parthenolide can be used for preparing a medicine for preventing and / or treating nicotine addiction.

[0006] TECHNICAL SCHEME The present application provides a use of parthenolide in preparation of a medicine for preventing and / or treating nicotine addiction.

[0007] The medicine comprises parthenolide and a pharmaceutically acceptable carrier or excipient.

[0008] The excipient comprises one or more of an emulsifier, an antioxidant, a wetting agent, a preservative, a diluent, a disintegrant or a binder.

[0009] The emulsifier is at least one selected from Tween, Span, glycerol fatty acid ester, pectin, agar, sodium alginate or silicon dioxide.

[0010] The antioxidant is at least one selected from ascorbic acid, sulfite, bisulfite, gallic acid and the like.

[0011] The wetting agent is selected from at least one of water or ethanol.

[0012] The preservative is selected from at least one of benzoic acid and its salts, sorbic acid and its salts, or nipagin.

[0013] The diluent is selected from at least one of starches, sugars, celluloses, or inorganic salts.

[0014] The disintegrant is selected from at least one of starch, sodium carboxymethyl starch, cross-linked polyplasdone, low-substituted hydroxypropyl cellulose, or cross-linked polyvinylpyrrolidone.

[0015] The binder is selected from at least one of starch paste, sodium carboxymethyl cellulose, povidone, hydroxypropyl cellulose, methyl cellulose, or ethyl cellulose.

[0016] The dosage form of the drug is granules, tablets, capsules, suspensions, oral solutions, or injections.

[0017] The effective dose of the parthenolide is 10-40 mg / kg.

[0018] The drug of the present application can be administered in various known ways, such as orally, by injection, or by inhalation spray.

[0019] The drug of the present application can be administered alone or in combination with other drugs. The oral composition can be in any orally acceptable dosage form, including but not limited to granules, tablets, capsules, suspensions, and solutions. The sterile injectable composition can be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. Pharmaceutically acceptable carriers and solvents that can be used include water, sodium chloride solution, and the like.

[0020] The drug of the present application can be prepared into a general preparation, or into a sustained-release preparation, a controlled-release preparation, a targeted preparation, and various microparticle drug delivery systems.

[0021] The actual dose level of the active ingredients in the drug of the present application can be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dose level will depend on a variety of factors including the route of administration, the time of administration, the rate of excretion, the duration of the treatment, other drugs, compounds, and / or materials used in combination with parthenolide, the age, sex, weight, general health condition, and prior medical history of the patient being treated, and similar factors well known in the medical arts.

[0022] The parthenolide is prepared as a drug for preventing and / or treating nicotine addiction as the only active ingredient.

[0023] The parthenolide is combined with other drugs to prepare a drug for preventing and / or treating nicotine addiction.

[0024] Firstly, the application uses conditioned place preference (CPP) combined with subcutaneous administration of nicotine solution (0.5 mg / kg) to establish a nicotine addiction mouse model, and intervenes by intragastrically administering different doses of parthenolide, and evaluates the addiction of the mice in the model group and the administration group through the CPP experiment.

[0025] Meanwhile, the application uses a 7-day continuous mouse jugular vein self-administration of nicotine solution (0.03 mg / kg / infusion) to evaluate the addiction of the mice in the model group and the administration group through the nicotine self-administration experiment.

[0026] The application uses conditioned place preference (CPP) to intragastrically administer different doses of parthenolide, and excludes the influence of parthenolide itself on the CPP score of the mice through the CPP experiment.

[0027] The application uses a 14-day continuous subcutaneous injection of nicotine solution (2 mg / kg), and injects the nicotine solution four times a day at 7:00, 12:00, 17:00 and 22:00, during which the treatment group is intragastrically administered different doses of parthenolide once a day for intervention, and the control group is intragastrically administered 0.5% CMC-Na. Then, the mice are placed in a feeding cage for normal feeding to establish a nicotine withdrawal mouse model, and 24 hours after the injection is stopped, the influence of parthenolide on the emotional symptoms of the mice in withdrawal is evaluated through open field test (OFT), elevated plus maze (EPM) and tail suspension test (TST).

[0028] The application intragastrically administers different doses of parthenolide to normal mice, and evaluates the influence of parthenolide on the natural reward preference, spontaneous motor ability and related behaviors of the mice through sucrose preference test (SPT), OFT and EPM experiments.

[0029] In addition, the application uses conditioned place preference (CPP) combined with subcutaneous administration of nicotine solution (0.5 mg / kg) to establish a nicotine addiction mouse model, then places the nicotine addiction model mouse in a feeding cage for normal feeding, natural withdrawal for 3 days, uses subcutaneous injection of a small dose of nicotine solution (0.1 mg / kg) for ignition to establish a nicotine relapse mouse model, and simultaneously gives different doses of parthenolide by gavage to intervene, and evaluates the effect of parthenolide on the nicotine relapse behavior of the mouse through the CPP experiment.

[0030] The parthenolide prevents the formation of nicotine-induced CPP of the mouse and the formation of nicotine forced drug seeking.

[0031] The parthenolide improves nicotine withdrawal symptoms and does not affect the spontaneous motor ability of the mouse.

[0032] The parthenolide prevents nicotine relapse.

[0033] Beneficial effects: The parthenolide disclosed in the application can be used as an active ingredient and applied to the prevention and / or treatment of nicotine addiction, which develops a new use of parthenolide and provides a new choice for preparing a medicine for preventing and / or treating nicotine addiction.

[0034] The parthenolide of the compound of the application prevents the formation of nicotine-induced CPP of the mouse by oral administration, can prevent nicotine addiction, significantly improves nicotine withdrawal symptoms and does not affect the spontaneous motor ability of the mouse. At the same time, the parthenolide of the application prevents nicotine relapse by oral administration. Therefore, the parthenolide of the application can be used for preparing a medicine for preventing and / or treating nicotine addiction. DETAILED DESCRIPTION

[0035] The technical solutions of the application will be described in detail below through specific embodiments, but the protection scope of the application is not limited to the embodiments.

[0036] If a specific technology or condition is not specified in the embodiments, the technology or condition described in the literature in the art or according to the product manual is used. If the manufacturer of the reagent or instrument is not specified, it is a conventional product that can be purchased through a regular channel.

[0037] In the following embodiments, the experimental methods are conventional methods unless otherwise specified. The test materials used in the following embodiments are commercially available products unless otherwise specified.

[0038] Parthenolide, content ≥98%, CAS No. 20554-84-1, Catalog No. XTY20250402, purchased from Jiangsu Boquan Biotechnology Co., Ltd., the dose was 10, 20, 40 mg / kg, and 0.5% CMC-Na solution was used to prepare 1, 2, 4 mg / ml parthenolide solution before use, and the volume of the drug was 0.1 ml / 10 g.

[0039] Preparation of nicotine stock solution: 28.5 mg of L-nicotine tartrate salt (equivalent to 10 mg of nicotine) was dissolved in 2 ml of sterile normal saline to prepare a stock solution containing 5 mg / ml of nicotine, which was prepared fresh every day.

[0040] Nicotine solution (0.05 mg / ml): 0.1 ml of nicotine stock solution was taken and added to 10 ml of sterile normal saline to obtain a 0.05 mg / ml nicotine solution, and the subcutaneous injection volume was 0.1 ml / 10 g.

[0041] Nicotine solution (0.03 mg / ml): 0.1 ml of nicotine stock solution was taken and added to 16.7 ml of sterile normal saline to obtain a 0.03 mg / ml nicotine solution. For intravenous injection (20 μl / infusion) in the self-administration experiment.

[0042] Nicotine solution (0.2 mg / ml): 6 mg of L-nicotine tartrate salt (equivalent to 2 mg of nicotine) was accurately weighed on an analytical balance and dissolved in 10 ml of sterile normal saline to prepare a solution containing 0.2 mg / ml of nicotine, and the subcutaneous injection volume was 0.1 ml / 10 g.

[0043] Nicotine solution (0.01 mg / ml): 0.1 ml of nicotine stock solution was taken and added to 50 ml of sterile normal saline to obtain a 0.01 mg / ml nicotine solution, and the subcutaneous injection volume was 0.1 ml / 10 g.

[0044] 0.9% Sodium Chloride Injection, Batch No. 2405228, produced by Jiangsu Huaian Shuanghe Pharmaceutical Co., Ltd.

[0045] Example 1 Effect of parthenolide on nicotine-induced addictive behavior in mice A nicotine addiction mouse model was established by combining conditioned place preference (CPP) experiment with subcutaneous administration of nicotine solution (0.5 mg / kg), and different doses of parthenolide were given by gavage to intervene, and the addictive behavior of mice in the model group and the drug administration group was evaluated by CPP experiment.

[0046] Experimental animals: male C57BL / 6J mice, body weight 20-25 g, production license number SCXK(SU)2024-0001, provided by Suzhou University of Health and Environmental Technology Co., Ltd.

[0047] Laboratory room temperature 20-22℃, relative humidity 40-60%, light 12h / day, cage breeding, 5 per cage, cage cleaning every five days.

[0048] Experimental grouping: mice were randomly divided into five groups according to body weight, namely normal control group (n=10), negative control group (n=12), parthenolide 10mg / kg group (n=12), parthenolide 20mg / kg group (n=12) and parthenolide 40mg / kg group (n=8).

[0049] Experimental method: (1) Conditioned place preference experiment: the experimental device is composed of two same size boxes (24cm×24cm×30cm) and a white relatively narrow buffer box (10cm×24cm×30cm), the gate at the middle connection can be opened or closed at will, the two boxes are striped and gray respectively, and a camera is installed above each box to record the residence time, movement distance and movement trajectory of the mouse in each box.

[0050] The experiment is divided into three stages: Pre-measurement stage: the mouse is slowly put into the CPP device from the middle buffer box, the gate is opened, and the residence time of the mouse in the three box chambers within 15 minutes is recorded, according to the residence time of the mouse in the striped room and the gray room, the initial preference room of the mouse is evaluated, and the non-initial preference room is used as the drug pairing room.

[0051] Training stage: in the 4-day training, the gate is closed to avoid the mouse back and forth. Train twice a day, at least 4 hours apart in the morning and afternoon. In the morning, the mouse is trained in the non-drug pairing room for 30 minutes after administration according to the grouping, and in the afternoon, the mouse is trained in the drug pairing room for 30 minutes after administration according to the grouping.

[0052] Post-measurement stage: open the gate and slowly put the mouse into the CPP device from the buffer box, let it freely move in the CPP device, and record the residence time of the mouse in each box chamber within 15 minutes, the total movement distance and movement trajectory of the mouse. According to the results of pre-measurement and post-measurement, the CPP score of each mouse is calculated.

[0053] CPP score = T (residence time of mouse in drug pairing room in post-measurement) - T (residence time of mouse in drug pairing room in pre-measurement) The experimental mice were trained for 4 days, 2 times / day (the administration schedule is shown in Table 1). The parthenolide treatment group of mice was given parthenolide (10, 20, and 40 mg / kg) by gavage 30 min before subcutaneous injection of nicotine solution (0.5 mg / kg), and the mice were placed in the corresponding box for training and learning for 30 min (the gate was closed to avoid shuttle).

[0054] The negative control group and the normal control group of mice were given 0.5% CMC-Na by gavage 30 min before the negative control group was subcutaneously injected with nicotine solution (0.5 mg / kg) and the normal control group was subcutaneously injected with normal saline, and the mice were placed in the corresponding box for training and learning for 30 min. The first day after training, the mice were tested by placing them in the middle box and allowing them to shuttle freely for 15 min. The behavior analysis software recorded the movement trajectory and the time spent in each box, and the data were collected by the DigBehv animal behavior analysis system. The ANY-maze software was used for analysis. The CPP score was calculated by comparing the previous test data to determine whether the mice changed their natural preference and developed addiction.

[0055] CPP score = T (time spent by mice in the drug-paired chamber in the post-test) - T (time spent by mice in the drug-paired chamber in the pre-test). The experimental data was processed using Graph Pad Prism 10.1.2. The experimental data results are shown in Table 2.

[0056] Table 1 Administration schedule of parthenolide in the CPP experiment of nicotine addiction model mice

[0057]

[0058] Note: Saline represents normal saline, and CMC-Na represents 0.5% carboxymethylcellulose sodium solution.

[0059] Table 2 Effect of parthenolide on CPP score of nicotine addiction model mice ± SEM

[0060] *P < 0.05, **P < 0.01, ***P < 0.001, Compared with the negative control group The experimental results showed that, in the CPP experiment, compared with the negative control group, the CPP score of mice given 10, 20, and 40 mg / kg parthenolide by gavage during the training period was significantly or extremely significantly decreased (P < 0.05 or P < 0.01, respectively) P < 0.05, P < 0.01, P < 0.001 ), as shown in Table 2. The experimental results showed that oral administration of parthenolide can prevent the formation of nicotine-induced CPP in mice and can prevent nicotine addiction in mice.

[0061] Example 2 Effect of parthenolide on nicotine-induced addiction behavior in mice The present application uses a 7-day mouse jugular vein self-administration of nicotine solution (0.03 mg / kg / infusion) to evaluate the model group and administration group mouse addiction through nicotine self-administration experiment.

[0062] Experimental animals: male C57BL / 6J mice, body weight 20-25 g, production license number SCXK(SU)2024-0001, provided by Suzhou Suda Health and Environmental Technology Research Co., Ltd.

[0063] Laboratory room temperature 20-22℃, relative humidity 40-60%, light 12h / day, cage breeding, cage cleaning every five days, single-cage feeding of mice after jugular vein cannulation surgery.

[0064] Experimental grouping: the mice were randomly divided into five groups according to body weight, namely normal control group (n=9), negative control group (n=8), parthenolide 10 mg / kg group (n=9), parthenolide 20 mg / kg group (n=8) and parthenolide 40 mg / kg group (n=10).

[0065] Experimental method: All behavioral experiments were performed during the light phase in an environmentally controlled box equipped with soundproofing. The self-administration device (Zhongshi Technology) was used, and each side of the single wall of the experimental box was equipped with a round hole, and a prompt light was configured above the round hole. The box was equipped with an indicator light. When the jugular vein was injected with drugs for self-administration, the catheter device was connected to the infusion pump (Zhongshi Technology). The experimental device was controlled and data were collected through the computer running the system.

[0066] In order to familiarize the mice with the self-administration environment, we conducted a three-day food shaping training during which the mice had to poke the effective round hole once to obtain food rewards (FR1 program, each time a 2mm diameter round sugar particle was given, the off period was 20s, and the training was performed for 60 minutes per day). After food training, the mice were anesthetized, the right jugular vein was observed on the right side of the mouse's neck and chest, the vein direction was identified, and a 2cm×2cm surgical area was marked on the right side of the midline for shaving. The subcutaneous superficial and deep fascia was separated with surgical scissors, and the muscle tissue was bluntly separated along the fiber direction with tissue scissors to find the right jugular vein. After determining the position of the right jugular vein, the direction of the blood flow was observed, and the main branch of the right jugular vein was found. Then, the fat tissue was separated with forceps, and two non-absorbing suture lines were marked below it for future use. During the process, the surgical site was intermittently moistened with normal saline. Then, the venous cannulation was performed, and the syringe connected to the catheter end was withdrawn. Whether the cannula position was correct was judged according to whether blood could be withdrawn. Successful blood return represented correct cannula position. The catheter (China Shanghai Yuan Industry) was threaded subcutaneously from the neck area to the dorsal side. Then, the incision was sutured, and the patency of the catheter was maintained by daily intravenous injection of heparin.

[0067] Mice were allowed to recover for 5 days post-surgery, and starting on the third day post-surgery, mice in the parthenolide 10, 20, and 40 mg / kg groups were gavaged with parthenolide, and the normal control and negative control groups were gavaged with CMC-Na solution. They then underwent the FR1 procedure for 7 days of nicotine self-administration, each lasting 2 hours. Active nose-poke valid holes triggered intravenous infusion of nicotine (20 μL, 0.03 mg / kg / infusion), with the chamber light turned off, accompanied by a 1-second cue light above the active port, and a 1-second audible tone. Inactive nose-poke did not trigger nicotine infusion, cue light, or tone. There was a 20-second refractory period after each nicotine infusion. The specific dosing is shown in Table 3. The experimental data was processed using Graph Pad Prism 10.1.2. The experimental data results are shown in Table 4.

[0068] Table 3 Dosing schedule for parthenolide intervention in nicotine addiction model mice self-administration experiment

[0069] Note: Saline represents normal saline, and CMC-Na represents 0.5% carboxymethylcellulose sodium solution.

[0070] Table 4 Effect of parthenolide on active nose-poke in nicotine addiction model mice self-administration experiment ± SEM

[0071] *P < 0.05, **P < 0.01, ***P < 0.001, Compared with the negative control group The experimental results show that, in the self-administration experiment, compared with the negative control group, gavage with 10, 20, and 40 mg / kg parthenolide significantly or extremely significantly reduced the active nose-poke of mice (P < 0.05 or P < 0.01) P < 0.05, P < 0.01, P < 0.001 , as shown in Table 4. The experimental results show that parthenolide can treat and / or prevent the compulsive drug-seeking behavior of mice for nicotine.

[0072] Example 3 Effect of parthenolide on conditioned place preference experiment in normal mice In order to rule out the influence of parthenolide addiction, this example gavaged normal mice with 10, 20, and 40 mg / kg parthenolide, and then evaluated the addiction of the dosed mice through the CPP experiment.

[0073] Experimental animals: male C57BL / 6J mice, weighing 20-25 g, production license number SCXK(Su)2024-0001, provided by Suzhou Sundai Health and Environmental Technology Research Co., Ltd. for quality testing.

[0074] Laboratory room temperature 20-22℃, relative humidity 40-60%, light 12h / day, cage breeding, 5 per cage, cage cleaning every five days.

[0075] Experimental grouping: mice were randomly divided into five groups according to body weight, namely normal control group, parthenolide 10, 20 and 40 mg / kg groups (n=10).

[0076] Experimental method: In the pre-test stage, the mice were placed in the middle buffer chamber, the gate was opened, and they were allowed to move freely for 15 min. The time spent and path in each chamber were recorded, and the preferred and non-preferred chambers were selected. The non-preferred chamber was set as the drug-paired chamber.

[0077] In the training stage, the experimental mice were trained for 4 days, 2 times / day (see Table 5 for drug administration). The experimental mice were placed in the corresponding chamber for 30 min of training and learning (the gate was closed to avoid shuttle).

[0078] In the post-test stage, the test was conducted on the first day after training. The mice were placed in the middle buffer chamber and allowed to move freely for 15 min. The behavior analysis software recorded the movement trajectory and the time spent in each chamber. The data were collected by the DigBehv animal behavior analysis system and analyzed using ANY-maze software. The CPP score was calculated by comparing the pre-test data to determine whether the mice changed their natural preference and developed addiction.

[0079] CPP score = T (time spent by mice in drug-paired chamber in post-test) - T (time spent by mice in drug-paired chamber in pre-test). The data were processed using Graph Pad Prism 10.1.2. The experimental data results are shown in Table 6.

[0080] Table 5 Drug administration schedule for the effect of parthenolide on CPP in normal mice

[0081] Note: CMC-Na represents 0.5% carboxymethylcellulose sodium solution.

[0082] Table 6 Effect of parthenolide on CPP score in normal mice ±SEM)

[0083] # P＞0.05， Compared with the normal control group The experimental results showed that in the CPP experiment, compared with the normal control group, the mice administered with 10, 20 and 40 mg / kg parthenolide during the training period had no significant difference in CPP score # P＞0.05), see Table 6. The experimental results show that parthenolide has no effect on the natural preference of mice.

[0084] Example 4 Effect of parthenolide on the withdrawal emotional symptoms of nicotine-addicted mice Experimental animals: male C57BL / 6J mice, body weight 20-25g, production license number SCXK(SU)2024-0001, provided by Suzhou University of Health and Environmental Technology Co., Ltd.

[0085] Laboratory room temperature 20-22℃, relative humidity 40-60%, light 12h / day, cage breeding, 5 per cage, cage cleaning every five days.

[0086] Experimental grouping: the mice were randomly divided into five groups according to body weight, namely normal control group, negative control group, parthenolide 10mg / kg, 20mg / kg and 40mg / kg treatment groups (n=10).

[0087] Experimental method: The negative control group and the parthenolide 10, 20 and 40mg / kg treatment groups were subcutaneously injected with nicotine 4 times a day, with a dose of 2mg / kg, while the normal control group received an equal amount of normal saline. The injection time started at 7:00 in the morning, followed by 12:00 and 17:00 in the afternoon, and the last one at 22:00 at night. At the same time, they were given a daily gavage. The normal control group and the negative control group were given a daily gavage of 0.5% CMC-Na solution, while the parthenolide 10, 20 and 40mg / kg treatment groups were given a daily gavage of parthenolide (mixed with 0.5% CMC-Na) for 14 days. After 24 hours of stopping nicotine injection, the mice were subjected to open field test (OFT), elevated plus maze test (EPM), and tail suspension test (TST).

[0088] OFT test: The test device consists of a blue plastic box (42cm x 42cm x 40cm), and data collection is performed by a Ji Liang DigBehv animal behavior analysis system. The mouse is placed in the center of the box and allowed to move freely. The movement trajectory within 5 minutes is recorded by a video analysis system, and the total movement distance and central stay time are analyzed. During the test, the environment is quiet and the light is uniform. After each mouse is tested, the excrement is cleaned up in time, and the inside of the box is wiped with 75% ethanol.

[0089] EPM test: The test device consists of two open arms (30 cm x 5 cm), two closed arms (30 cm x 5 cm x 15 cm), and is 40-55 cm high from the ground, and data collection is performed by the Ji Liang DigBehv animal behavior analysis system. At the beginning of the experiment, the mouse is placed in the maze from the central grid to the open arm, and the activity within 5 min is recorded, and the open arm residence time of the experimental mouse is analyzed. During the test, the environment is quiet, the light is uniform, and after each mouse test, the excrement is cleaned in time, and wiped with 75% alcohol.

[0090] TST test: TST is used to assess "behavioral despair", and the mouse's tail is fixed on the rod with tape to hang. One end of the tape is fixed on the horizontal rod to ensure that the distance between the mouse's nose and the device floor is about 20-25 cm. Video record the behavior for 5 minutes, and count the immobile time.

[0091] The experimental data were processed by Graph Pad Prism 10.1.2. The experimental data results are shown in Tables 7, 8, 9, and 10.

[0092] Table 7 Effect of parthenolide on OFT in nicotine withdrawal model mice ±SEM)

[0093] *P<0.05， # P＞0.05， Compared with the negative control group Table 8 Effect of parthenolide on OFT in nicotine withdrawal model mice ±SEM)

[0094] # P＞0.05， Compared with the negative control group Table 9 Effect of parthenolide on EPM in nicotine withdrawal model mice ±SEM)

[0095] *P < 0.05, **P < 0.01, ***P < 0.001, Compared with the negative control group Table 10 Effect of parthenolide on TST in nicotine withdrawal model mice ±SEM)

[0096] **P < 0.01, ***P < 0.001, Compared with the negative control group The experimental results show that, in the OFT, compared with the negative control group, the mice administered with 10, 20, 40 mg / kg parthenolide by gavage have an increased central residence time, among which the mice administered with 10 mg / kg parthenolide by gavage have the most significant increase in central residence time P＜0.05 , but have no effect on the total distance of mouse movement P＞0.05 , as shown in Tables 7 and 8.

[0097] In the EPM experiment, compared with the negative control group, the mice administered with 10, 20, 40 mg / kg parthenolide by gavage have a significantly increased time of staying in the open arm P < 0.05, P < 0.01, P < 0.001 , as shown in Table 9. In the TST experiment, compared with the negative control group, the mice administered with 10, 20, 40 mg / kg parthenolide by gavage have a significantly or extremely significantly reduced immobility time P < 0.05, P < 0.01, 0.01，P＜0.001 , as shown in Table 10. The above experimental results show that parthenolide can significantly improve the anxiety and depression-like behaviors of mice after nicotine withdrawal, and does not affect the spontaneous activity of mice.

[0098] Example 5: Effect of parthenolide on the open field test of normal mice Experimental animals: male C57BL / 6J mice, weighing 20-25 g, production license number SCXK(Su)2024-0001, provided by Suzhou Sundai Health and Environmental Technology Research Co., Ltd. for quality testing.

[0099] Laboratory room temperature 20-22℃, relative humidity 40-60%, light 12h / day, cage breeding, 4 mice per cage, cage cleaning every five days.

[0100] Experimental grouping: the mice were randomly divided into four groups according to body weight, i.e. normal control group, parthenolide 10, 20 and 40 mg / kg treatment groups (n=8).

[0101] Experimental method: According to the grouping, the mice were given solvent (0.5% CMC-Na solution) or parthenolide by gavage, and the OFT was performed 30 min later (experimental method same as Example 4). The movement trajectory of the mice within 5 min was recorded by a video analysis acquisition system, and the total distance of mouse movement and the central residence time were obtained by system analysis. The experimental data were processed by Graph Pad Prism 10.1.2. The experimental data results are shown in Tables 11 and 12.

[0102] Table 11: Effect of parthenolide on the OFT of mice ±SEM

[0103] #P＞0.05， Compared with the normal control group Table 12 Effect of parthenolide on OFT in mice ±SEM

[0104] # P＞0.05， Compared with the normal control group The experimental results show that, in OFT, compared with the solvent control group (i.e. the normal control group), intragastrically administered 10, 20, 40 mg / kg parthenolide has no significant effect on the total distance of movement and the central residence time of mice P＞0.05 , see Tables 11 and 12. This shows that different doses of parthenolide do not affect the autonomous movement ability of normal mice.

[0105] Example 6 Effect of parthenolide on the elevated plus maze test of normal mice Experimental animals: male C57BL / 6J mice, body weight 20-25 g, production license number SCXK (Su) 2024-0001, provided by Suzhou Sundai Health and Environment Technology Research Co., Ltd. for quality testing.

[0106] Laboratory room temperature 20-22℃, relative humidity 40-60%, light 12h / day, cage breeding, 4 mice per cage, cage cleaning every five days.

[0107] Experimental grouping: the mice were randomly divided into four groups according to body weight, i.e. normal control group, parthenolide 10, 20 and 40 mg / kg treatment groups (n=8).

[0108] Experimental method: According to the grouping, the mice were given solvent (0.5% CMC-Na solution) or parthenolide by gavage, and EPM was performed 30 min later (experimental method same as Example 4). The movement trajectory of the mice within 5 min was recorded using a video analysis acquisition system, and the open arm residence time of the mice was obtained by system analysis.

[0109] The experimental data were processed by Graph Pad Prism 10.1.2. The experimental data results are shown in Table 13.

[0110] Table 13 Effect of parthenolide on EPM in mice ±SEM

[0111] # P＞0.05， Compared with the normal control group The experimental results showed that in the EPM, compared with the normal control group, intragastric administration of 10, 20, and 40 mg / kg parthenolide had no effect on the time that mice stayed in the open arms ( P＞0.05 ), as shown in Table 13.

[0112] Example 7 Effect of parthenolide on the sucrose preference test in normal mice Experimental animals: Male C57BL / 6J mice, weighing 20 - 25 g, production license number SCXK (Su) 2024 - 0001, provided by Suzhou University of Science and Technology Health and Environmental Technology Research Co., Ltd. for quality inspection.

[0113] The laboratory room temperature was 20 - 22 °C, the relative humidity was 40 - 60%, the light was 12 h / day, and they were cage - raised, with 5 mice in each cage. The cage was cleaned once every five days.

[0114] Experimental grouping: The mice were randomly divided into four groups according to body weight: a normal control group, and parthenolide treatment groups of 10, 20, and 40 mg / kg (n = 10).

[0115] Experimental method: The sucrose preference test (SPT) is usually used to simulate anhedonia or the inability to feel pleasure. Here, the mice were individually housed and accustomed to two water bottles for 2 days. After 24 hours of water deprivation, the mice were intragastrically administered with vehicle (0.5% CMC - Na solution) or parthenolide according to the grouping, and the mice were exposed to one bottle of 1.5% sucrose and one bottle of water for 12 hours. During the continuous 12 - hour test, the bottle positions were changed every 6 hours. The total liquid consumption, sucrose consumption, and pure water consumption were recorded, and the sucrose preference index of each mouse was calculated, that is: Sucrose preference index = (sucrose consumption / total liquid consumption) × 100%.

[0116] The experimental data were all processed by Graph Pad Prism 10.1.2. The experimental data results are shown in Table 14.

[0117] Table 14 Effect of parthenolide on SPT in mice ( ±SEM)

[0118] # P＞0.05， Compared with the normal control group The experimental results showed that in the SPT, compared with the normal control group, intragastric administration of 10, 20, and 40 mg / kg parthenolide had no effect on the sucrose preference index of mice ( P＞0.05 ), as shown in Table 14. This indicates that different doses of parthenolide do not affect the natural reward behavior of experimental mice under physiological conditions.

[0119] Effect of parthenolide on relapse behavior of nicotine addicted mice Experimental animals: male C57BL / 6J mice, weighing 20-25 g, production license number SCXK(SU)2024-0001, provided by Suzhou University Health and Environmental Technology Research Co., Ltd.

[0120] Laboratory room temperature 20-22℃, relative humidity 40-60%, light 12h / day, cage breeding, 4-5 per cage, cage cleaning every five days.

[0121] Experimental grouping: mice were randomly divided into five groups according to body weight, normal control group (n=10), negative control group (n=10), parthenolide 10, 20 and 40mg / kg treatment groups (n=9).

[0122] Experimental method: In the pre-test stage, the mice were placed in the middle buffer chamber, the gate was opened, and they were allowed to move freely for 15 min, the time spent and path in each chamber were recorded, the preferred chamber and non-preferred chamber were selected, and the non-preferred chamber was set as the drug-paired chamber.

[0123] In the training stage, the experimental mice were trained for 4 days, 2 times / day (dosing schedule see Table 15), subcutaneous injection of nicotine solution (0.5mg / kg), and the experimental mice were placed in the corresponding chamber for training and learning for 30 min (the gate was closed to avoid shuttle).

[0124] In the post-test stage, the test was performed on the first day after training, the mice were placed in the middle chamber and allowed to shuttle freely for 15 min, the behavior analysis software recorded the movement trajectory and the time spent in each chamber, the CPP score was calculated by comparing the pre-test data, and whether the mice were addicted was determined.

[0125] CPP score = T (time spent by mice in drug-paired chamber in post-test) - T (time spent by mice in drug-paired chamber in pre-test) After confirming the addiction of the mice, the mice were placed in the breeding cage for normal feeding and natural extinction, on the 3rd day of extinction, the conditioned place preference experiment (CPP experiment) was performed. Before the experiment, nicotine was dissolved in sterile saline, the pH was adjusted to 7.4, the mice were subcutaneously injected with nicotine solution (0.1mg / kg) for ignition, the normal control group and the negative control group were given the same amount of solvent by gavage 30 min before the experiment, the parthenolide groups were given different doses of parthenolide solution by gavage 30 min before the experiment, except for the normal control group which was subcutaneously injected with the same amount of sterile saline, the mice in the other groups were subcutaneously injected with nicotine solution (0.1mg / kg) immediately after which the CPP experiment was performed.

[0126] All experimental data were processed using Graph Pad Prism 10.1.2. The experimental results are shown in Tables 16 and 17.

[0127] Table 15 Dosing schedule for the experiment of ternolactone intervention on relapse behavior in nicotine-addicted mice

[0128] Note: Saline represents physiological saline, and CMC-Na represents 0.5% sodium carboxymethyl cellulose solution.

[0129] Table 16 CPP scores of mice in a nicotine addiction model established by subcutaneous nicotine injection ( ±SEM)

[0130] *P < 0.05, **P < 0.01, Compared with the normal control group Table 17 Effect of ternolactone on CPP score in nicotine relapse model mice ( ±SEM)

[0131] *P < 0.05, **P < 0.01, Compared with the negative control group Experimental results showed that in the CPP experiment of constructing a nicotine addiction model mouse, compared with the normal control group, the CPP scores of mice in each group were significantly increased without the administration of parthenolide. P＜0.01 (See Table 16). The experimental results show that subcutaneous injection of 0.5 mg / kg nicotine successfully established nicotine addiction in mice. During the relapse phase, compared with the negative control group, gavage administration of 20 and 40 mg / kg parthenolide during the training period significantly or extremely significantly reduced the CPP score of the mice. P < 0.05, P < 0.01, 0.01 ), oral administration of 10 mg / kg parthenolide decreased the CPP score in mice, but the difference was not statistically significant. P＞0.05 (See Table 17). The experimental results showed that 10, 20, and 40 mg / kg of parthenolide reduced nicotine relapse behavior in mice.

[0132] These results indicate that parthenolide can be used to prepare drugs for the prevention and / or treatment of nicotine addiction.

[0133] As described above, although the invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the invention itself. Various changes in form and detail may be made without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. Use of parthenolide in the preparation of a medicament for preventing and / or treating nicotine addiction.

2. Use according to claim 1, characterized in that, The medicament comprises parthenolide and a pharmaceutically acceptable carrier or excipient.

3. Use according to claim 1, characterized in that, The dosage form of the medicament is granules, tablets, capsules, suspensions, oral solutions or injections.

4. Use according to claim 1, characterized in that, The effective dose of parthenolide is 10-40 mg / kg.

5. Use according to claim 1, characterized in that, The parthenolide is used as the only active ingredient in the preparation of a medicament for preventing and / or treating nicotine addiction.

6. Use according to claim 1, characterized in that, The parthenolide is used in combination with other drugs in the preparation of a medicament for preventing and / or treating nicotine addiction.

7. Use according to claim 1, characterized in that, The parthenolide prevents the formation of nicotine-induced CPP in mice and the formation of nicotine forced seeking.

8. Use according to claim 1, characterized in that, The parthenolide improves nicotine withdrawal symptoms and does not affect the spontaneous motor ability of mice.

9. Use according to claim 1, characterized in that, The parthenolide prevents nicotine relapse.

Images