Pharmaceutical composition for treating alcohol dependence as well as preparation method and application thereof

Abstract

The invention discloses a pharmaceutical composition for treating alcohol dependence and a preparation method and application thereof. The pharmaceutical composition is prepared from 100 to 1,000 mg of ascorbic acid, 10 to 40 mg of pyrroloquinoline quinone and pharmaceutically acceptable auxiliary materials. According to the pharmaceutical composition provided by the invention, ascorbic acid and pyrroloquinoline quinone are combined for application, so that the alcohol preference rate of mice is remarkably reduced. In a mouse alcohol dependence model, a mouse taking the pharmaceutical composition shows obvious reduction of alcohol intake, and the alcohol dependence is lower. The pharmaceutical composition provided by the invention has the potential of inhibiting alcohol craving and relieving alcohol addiction behaviors, and can effectively intervene in formation and maintenance of alcohol dependence.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly relates to a pharmaceutical composition for treating alcohol dependence, its preparation method and application. Background Art

[0002] Alcohol Dependence refers to a dual physical and psychological dependence state that individuals develop after long-term and repeated alcohol intake. Its main manifestations include a strong craving for alcohol, increased tolerance, decreased control, and obvious withdrawal symptoms when alcohol intake is stopped or reduced. Alcohol Dependence has been listed by the World Health Organization as one of the mental disorders that seriously endanger human health, featuring high relapse rate, high disability rate, and high social burden.

[0003] Currently, the drugs clinically used for treating alcohol dependence mainly include abstinence drugs (such as disulfiram), anti-craving drugs (such as naltrexone), withdrawal relief drugs (such as benzodiazepines), etc. However, these drugs generally have problems such as limited efficacy, large side effects, poor compliance, and are prone to drug tolerance or dependence, making it difficult to meet the clinical needs of long-term and systematic intervention. Therefore, developing a new type of pharmaceutical composition with high safety, good compliance, and multi-target synergistic action mechanism has become an important direction in current drug research and development. Summary of the Invention

[0004] Aiming at the defects in the prior art, the present invention provides a pharmaceutical composition for treating alcohol dependence, its preparation method and application.

[0005] The present invention provides a pharmaceutical composition for treating alcohol dependence and / or alcohol addiction, comprising 100 - 1000 mg of ascorbic acid, 10 - 40 mg of pyrroloquinoline quinone, and pharmaceutically acceptable excipients.

[0006] The pharmaceutical composition of the present invention combines pyrroloquinoline quinone with ascorbic acid and is applicable to the recovery of nervous system damage caused by alcohol dependence or alcohol addiction. In the present invention, the addition of pyrroloquinoline quinone not only strengthens the antioxidant and neuroprotective effects of ascorbic acid, but also may enhance its comprehensive ability to repair nerve damage and intervene in withdrawal symptoms by improving cellular energy metabolism, reducing inflammation, stabilizing neural networks, etc. The combined use is expected to more effectively reduce the alcohol preference rate of mice, reduce alcohol intake, and significantly alleviate withdrawal reactions.

[0007] In some embodiments, the excipients include any one or more of lecithin, soybean phospholipid, lactose, starch, gelatin, calcium carbonate, calcium phosphate, surfactant, sweetener, filler, binder, cellulose derivative, lubricant.

[0008] The above excipients are all commonly used pharmaceutical excipients included in the pharmacopoeia, and the pharmaceutically acceptable excipients described in the present invention are not limited thereto.

[0009] In some embodiments, the mass ratio of ascorbic acid to pyrroloquinoline quinone is (10~20):1. The above mass ratio range helps to optimize the combination ratio, exert the best neuroprotective and antioxidant effects, and at the same time reduce unnecessary ingredient waste or side effects.

[0010] In some embodiments, the pharmaceutical composition is an oral preparation. The active ingredients ascorbic acid and pyrroloquinoline quinone of the present invention can both stably exist in the oral preparation, which is convenient for long-term administration or withdrawal intervention.

[0011] In some embodiments, the oral preparation is any one of oral liquid, solid beverage, granule, capsule, tablet, effervescent tablet and pill. The dosage form of the pharmaceutical composition of the present invention has strong flexibility and can meet the needs of different populations and clinical applications.

[0012] In some embodiments, after the pharmaceutical composition is soaked in simulated gastric juice in vitro for 2 hours, the cumulative release rates of ascorbic acid and pyrroloquinoline quinone are both not less than 85%.

[0013] In some embodiments, the pH value of the pharmaceutical composition is 5.5~7.0. The pH value has an important influence on the drug stability, dissolution behavior and taste. The pharmaceutical composition within this pH range helps to enhance solubility, reduce gastric irritation and ensure the chemical stability of the ingredients.

[0014] The present invention also provides a preparation method of the pharmaceutical composition, comprising the following steps: S1: Weigh ascorbic acid and pyrroloquinoline quinone by weight and mix them evenly at a low speed in a dry mixer; S2: Add pharmaceutically acceptable excipients and prepare an oral preparation according to the selected dosage form.

[0015] The present invention also provides the application of the pharmaceutical composition in the preparation of a drug for treating alcohol addiction and / or alcohol dependence. This composition has been verified by animal experiments to effectively reduce the alcohol preference rate and relieve addiction symptoms, and is applicable to treatment stages such as addiction intervention, chronic dependence, and alleviating withdrawal reactions.

[0016] The present invention also provides the application of the pharmaceutical composition in the preparation of a drug for alcohol relapse intervention or reducing alcohol intake. This composition can relieve anxiety, tremors, the desire for impulsive drinking, etc., has the potential for adjuvant intervention, and can be used as a maintenance period drug or a supplementary drug for preventing relapse to extend the withdrawal effect.

[0017] In summary, compared with the prior art, the present invention has achieved the following technical effects: 1. The pharmaceutical composition provided by the present invention, through the combined application of ascorbic acid and pyrroloquinoline quinone, significantly reduces the alcohol preference rate of mice. In the alcohol dependence model of mice, the mice taking this pharmaceutical composition show a significant reduction in alcohol intake and exhibit lower alcohol dependence compared to the control group mice. This indicates that the pharmaceutical composition of the present invention has the potential to inhibit alcohol craving and alleviate alcohol addiction behavior, and can effectively intervene in the formation and maintenance of alcohol dependence.

[0018] 2. The pharmaceutical composition of the present invention can significantly alleviate various withdrawal symptoms that occur during the withdrawal period in alcohol-addicted mice. Evaluated by a standard withdrawal scoring scale, the mice using this composition show significantly lower withdrawal symptom scores, proving that the pharmaceutical composition has a positive effect in alleviating the withdrawal reaction of alcohol-dependent patients and can help reduce the discomfort and relapse risk caused by alcohol discontinuation.

[0019] 3. The pharmaceutical composition of the present invention, by combining pyrroloquinoline quinone and ascorbic acid, significantly enhances the therapeutic effect of ascorbic acid. It not only improves the efficacy of ascorbic acid but also provides a more efficient and stable treatment plan for the pharmaceutical treatment of alcohol dependence. Detailed implementation manners

[0020] In order to enable those skilled in the art to better understand the solution of the present invention, the technical solutions in the embodiments of the present invention are clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0021] The experimental methods used in the following embodiments are all conventional methods unless otherwise specified. The materials, reagents, etc. used can be obtained from commercial channels unless otherwise specified.

[0022] Example 1 (1) Raw material weighing and mixing: Weigh 500 mg of ascorbic acid and 30 mg of pyrroloquinoline quinone, and put them into a dry mixer, and stir evenly at a low speed until the ascorbic acid and pyrroloquinoline quinone are fully mixed. Keep the temperature below 40°C in this step to avoid the degradation of ascorbic acid.

[0023] (2) Adding excipients: Add 100 mg of lactose and 60 mg of calcium carbonate to the mixture of ascorbic acid and pyrroloquinoline quinone, and continue to stir under low-speed mixing conditions to ensure that all components are evenly distributed.

[0024] (3) Granulation and shaping: Using the wet granulation technique, the mixture is mixed with 3% hydroxypropyl methylcellulose binder to form granules. The granules are dried in a dryer until the water content does not exceed 5%.

[0025] (4)Tablet pressing of the preparation: The dried granules are pressed into tablets. Each tablet contains 500 mg of ascorbic acid and 30 mg of pyrroloquinoline quinone. The equipment used is a tablet press, and the initial pressing pressure for tablet pressing is set at 3 kN, and the main pressing (forming) pressure is set at 8 kN.

[0026] (5)pH measurement: Take 0.5 g of tablets, add them to a beaker, add 100 mL of deionized water, place them in a water bath at 37 ± 0.5 °C, stir magnetically, and use a pH meter to measure the pH value of the supernatant. The pH value of the drug combination tablet solution in this example is 5.6.

[0027] Example 2 The difference from Example 1 is that in the raw materials, ascorbic acid is 100 mg and pyrroloquinoline quinone is 10 mg.

[0028] The pH value of the drug combination tablet solution in this example is 6.1.

[0029] Example 3 The difference from Example 1 is that in the raw materials, ascorbic acid is 1000 mg and pyrroloquinoline quinone is 40 mg.

[0030] The pH value of the drug combination tablet solution in this example is 5.0.

[0031] Example 4 The difference from Example 1 is that the excipients are 50 mg of lecithin, 40 mg of soybean phospholipid, and 30 mg of gelatin.

[0032] The pH value of the drug combination tablet solution in this example is 6.2.

[0033] Comparative Example 1 The difference from Example 1 is that in the raw materials, only 500 mg of ascorbic acid is included.

[0034] The pH value of the drug combination tablet solution in this comparative example is 5.4.

[0035] Comparative Example 2 The difference from Example 1 is that in the raw materials, ascorbic acid is 50 mg and pyrroloquinoline quinone is 30 mg.

[0036] The pH value of the drug combination tablet solution in this comparative example is 5.6.

[0037] Comparative Example 3 The difference from Example 1 is that in the raw materials, ascorbic acid is 1500 mg and pyrroloquinoline quinone is 30 mg.

[0038] The pH value of the drug combination tablet solution in this comparative example is 4.5.

[0039] Comparative Example 4 The difference from Example 1 is that in the raw materials, ascorbic acid is 500 mg and pyrroloquinoline quinone is 5 mg.

[0040] The pH value of the drug combination tablet solution in this comparative example is 5.4.

[0041] Comparative Example 5 The difference from Example 1 is that in the raw materials, ascorbic acid is 500 mg and pyrroloquinoline quinone is 80 mg.

[0042] The pH value of the drug combination tablet solution in this comparative example is 5.3.

[0043] Comparative Example 6 The difference from Example 1 is that it is made into an injection, and the preparation method is as follows: Ascorbic acid and pyrroloquinoline quinone are weighed separately under sterile conditions and set aside. All excipients should meet the requirements for injection grade and be sterilized in advance under appropriate conditions.

[0044] Take a certain volume of injection water in the clean area, add glycerol, and stir until completely dissolved; add pyrroloquinoline quinone and ascorbic acid to this solution in sequence, and stir slowly until dissolved. Ascorbic acid needs to be operated under light protection to prevent oxidation; detect and adjust the pH of the solution to between 5.5 and 6.5; Use a 0.22 μm sterile filter for sterile filtration, filter into a sterilized injection vial, and seal immediately.

[0045] Drug dissolution experiment (1) Put the tablets prepared in Examples 1 to 4 into a rotating basket. At least 6 tablets / 6 granules should be used for each dissolution test sample to ensure statistical validity.

[0046] (2) Preparation of dissolution medium Prepare 0.1N hydrochloric acid solution and adjust the pH to 1.2 to simulate the acidic environment of gastric juice. Heat the solution to 37 ± 0.5 °C and place it in a dissolution tester.

[0047] (3) Dissolution test Place the rotating basket containing the drug sample into simulated gastric fluid and set the dissolution tester rotation speed to 50 rpm. During the dissolution process, take a 0.5 mL sample at regular intervals and filter the sample to remove suspended matter. The sampling time points are as follows: 0 hours (initial sample when adding the solution), 30 minutes, 1 hour, 1.5 hours, 2 hours.

[0048] (4) Sample treatment and analysis After each sampling, replenish the lost volume with 0.1N hydrochloric acid solution to keep the solution volume constant. Filter the sampled solution through a 0.45 μm filter membrane for quantitative analysis preparation.

[0049] Use an ultraviolet spectrophotometer to detect the concentrations of ascorbic acid (250 nm) and pyrroloquinoline quinone (280 nm), and calculate the drug release amount according to the standard curve.

[0050] (5) Cumulative release rate calculation Based on the sample concentrations at different time points, calculate the cumulative release amount of the drug in simulated gastric fluid in vitro. The cumulative release rate formula is: ; Where: R(t) is the cumulative release rate at time t ; C t is the drug concentration at time t ; V is the volume of the dissolution medium; C 0 is the initial drug concentration; M is the total mass of the drug.

[0051] With time as the abscissa and the cumulative release rate as the ordinate, plot the drug dissolution curve. Calculate the cumulative release rate of ascorbic acid and pyrroloquinoline quinone after 2 hours and compare it with the expected value to judge whether the drug release characteristics meet the expectations.

[0052] (6) Result requirements Cumulative release rate: After soaking in simulated gastric fluid in vitro for 2 hours, the cumulative release rate of ascorbic acid and pyrroloquinoline quinone should be not less than 85% to ensure good dissolution in the gastrointestinal tract.

[0053] (7) Test results According to the calculation results, after soaking the tablets prepared in Examples 1 - 4 in simulated gastric fluid in vitro for 2 hours, the cumulative release rates of ascorbic acid and pyrroloquinoline quinone are shown in Table 1: Table 1 Cumulative release rates of ascorbic acid and pyrroloquinoline quinone (2 hours)

[0054] As shown in Table 1, the cumulative release rates of ascorbic acid and pyrroloquinoline quinone in the tablets prepared in Examples 1 to 4 were greater than 85%, indicating that the pharmaceutical composition prepared by the present invention has good dissolution in the gastrointestinal tract.

[0055] Pharmacodynamic experiment Test Example 1 Establishment of Dependence and Addiction Mouse Models 1. Establishment of Alcohol Intake Model After adaptively feeding 10-week-old clean-grade C57BL / 6J male mice for one week, an alcohol intake model was established. The specific steps are shown in Table 2: Table 2 Gradually Establishing the Willingness to Drink during the Induction Period (Days 1 - 6)

[0056] Through the steps in Table 2, the drinking preference was gradually established and the taste of alcohol was adapted. Subsequently, from Day 7 to Day 21, the free-drinking (two-bottle method, one bottle containing 20% alcohol and the other bottle containing pure water, and the intake was recorded) drinking method was adopted. The mice were weighed and replenished with fluid at a fixed time in the morning, and gradually showed a preference for alcohol (alcohol intake > pure water intake) and behavioral dependence.

[0057] Methods for Evaluating Dependence and Addiction: Alcohol Preference Test (Two-Bottle Method) Weigh the two bottles of drinking water before and after each day.

[0058] Calculation formula: ; When the preference rate > 70%, it indicates an obvious tendency of alcohol addiction.

[0059] 2. Withdrawal Test (24 Hours of Alcohol Withdrawal) Observed symptoms: tremors, restlessness, scratching, jumping, increased aggression, etc.; The "withdrawal scoring scale" can be used for scoring, such as scoring according to the frequency of occurrence and severity (0 - 3 points); Obvious behavioral changes + high preference rate = successful model establishment.

[0060] Table 3 Withdrawal Scoring Criteria

[0061] Each mouse was observed for 5 minutes and scored according to the withdrawal scoring scale in Table 3. The scoring results are shown in Table 4 for reference: Table 4 Interpretation of Scoring Results

[0062] Before and after each test, the experimental conditions should be unified to avoid the influence of environmental stimulation differences on behavior. Finally, mice with an alcohol preference rate greater than 70% and a withdrawal score greater than 15 points are selected as the alcohol addiction model mice.

[0063] Test Example 2 Effect of the pharmaceutical composition on the alcohol preference of addicted mice The mice successfully established as models were randomly divided into a model group and a treatment group, with 6 mice in each group. The treatment group was continuously given the tablets prepared in Examples 1 to 4 and Comparative Examples 1 to 5 by gavage for 3 days respectively, and the injection prepared in Comparative Example 6 was administered by intraperitoneal injection at a dose of 200 mg / kg. The model group was injected with an equal amount of normal saline.

[0064] After the administration, the alcohol preference test (two-bottle method) in Test Example 1 was repeated for each group of mice, and the alcohol preference rate of the mice was calculated as shown in Table 5: Table 5 Alcohol preference rate (n = 6, %)

[0065] Among them, compared with the model group, **P < 0.01, *P < 0.01.

[0066] The results in Table 5 show that compared with the model group and the treatment groups of Comparative Examples 1 to 6, the alcohol preference rate of the treatment groups of Examples 1 to 4 was significantly reduced, indicating that the drugs prepared in Examples 1 to 4 of the present invention can significantly reduce the alcohol preference rate of alcohol-addicted mice. Compared with Example 1, the alcohol preference rate of Comparative Example 6 was significantly lower than that of the model group (P < 0.01), indicating that the pharmaceutical composition of the present invention is suitable for oral administration, and the oral administration method may be able to more effectively regulate and control the key centers of reward and drinking behavior, thereby affecting the alcohol preference rate of mice.

[0067] Test Example 3 Effect of the pharmaceutical composition on the withdrawal reaction of addicted mice After the above model group and treatment group mice stopped drinking for 24 hours, a withdrawal test was carried out, and the scoring situation of each group of mice was counted as shown in Table 6: Table 6 Statistical results of withdrawal test scores (points)

[0068] Among them, compared with the model group, **P < 0.01, *P < 0.01.

[0069] The results in Table 6 show that compared with the model group and the treatment groups of Comparative Examples 1 to 6, the withdrawal scores of the treatment groups of Examples 1 to 4 were significantly reduced, indicating that the drugs prepared in Examples 1 to 4 of the present invention can relieve the withdrawal reaction of alcohol-addicted mice in a short time.

[0070] Test Example 4 Nissl staining of neurons in the amygdala of addicted mice Nissl granules are basophilic substances present in the cytoplasm of neurons and are widely distributed in various types of neurons. Their main functions include: synthesizing structural proteins for updating cell organelles, synthesizing enzymes required for neurotransmitters, and peptide neuromodulators. The shape, number, and distribution of Nissl granules vary depending on the type of neuron. In neurons with active metabolic functions, the content of Nissl granules is particularly abundant. When neurons are damaged or over-fatigued, Nissl granules may decrease, disintegrate, or even completely disappear. However, during the recovery process of neurons from damage or fatigue, Nissl granules reappear and gradually increase, eventually returning to normal levels. Therefore, Nissl granules can be used as an important indicator to measure the functional state of neurons.

[0071] Referring to the administration and grouping in Test Example 1, after the administration was completed, the mice were sacrificed, and the brains were quickly removed and fixed in 10% formaldehyde solution, followed by conventional dehydration and embedding. The paraffin sections were 5 μm thick, and were conventionally dewaxed to water. They were stained with Nissl staining solution (Nissl Stain, toluidine blue method) in an incubator at 50 - 60 °C for 20 - 40 min. They were slightly washed with distilled water. They were quickly differentiated with 95% ethanol, dehydrated with absolute ethanol, cleared with xylene, and sealed with neutral gum. The pathological changes of neurons were observed under a light microscope, and the number of activated neurons was counted. The results are shown in Table 7.

[0072] Table 7 Number of activated neurons (pcs)

[0073] Among them, compared with the model group, **P < 0.01, *P < 0.01.

[0074] The cell body of a neuron contains a large cell nucleus with a folded nuclear membrane, sparsely distributed chromatin, and a distinct nucleolus. Nissl granules in the cytoplasm are its prominent feature. These granules represent rough endoplasmic reticulum and exhibit a specific speckled basophilic appearance in many neurons. The number and distribution of Nissl granules change with the physiological state of the neuron, and it is a key region for protein synthesis within the neuron. When the neuron is stimulated, the number of Nissl granules in the cell body changes.

[0075] The statistics of the number of neurons are shown in Table 7. After the brain neurons of the mice in the model group were stimulated by ethanol, the number of Nissl granules increased significantly, and the arrangement became more compact, indicating that the brain neurons of the mice were in an abnormally active state at this time. In contrast, in the experimental groups treated with inhibitory drugs, the effect of ethanol on the brain neurons of the mice was alleviated to a certain extent. In the brain neurons of these mice, the number of Nissl granules decreased, and the arrangement became relatively sparse. This shows that the drug compositions prepared in Examples 1 - 4 can effectively reduce the adverse effects of alcohol on neurons.

[0076] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A pharmaceutical composition for treating alcohol dependence and / or alcohol addiction, characterized in that: The invention comprises 100-1000 mg ascorbic acid, 10-40 mg pyrroloquinoline quinone and pharmaceutically acceptable excipients.

2. The pharmaceutical composition according to claim 1, characterized in that The auxiliary materials include any one or more of lecithin, soybean lecithin, lactose, starch, gelatin, calcium carbonate, calcium phosphate, surfactant, sweetener, filler, adhesive, cellulose derivative, and lubricant.

3. The pharmaceutical composition according to claim 1, characterized in that The mass ratio of ascorbic acid to pyrroloquinoline quinone is (10-20):

1.

4. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition is an oral preparation.

5. The pharmaceutical composition according to claim 4, characterized in that The oral preparation is any one of oral liquid, solid beverage, granule, capsule, tablet, effervescent tablet and pill.

6. The pharmaceutical composition according to claim 1, characterized in that After the pharmaceutical composition is immersed in simulated gastric fluid in vitro for 2 hours, the cumulative release rates of ascorbic acid and pyrroloquinoline quinone are not less than 85%.

7. The pharmaceutical composition according to claim 1, characterized in that The pH value of the pharmaceutical composition is 5.5-7.

0.

8. The method for preparing the pharmaceutical composition according to any one of claims 1 to 7, characterized in that: The steps include: S1: Weigh ascorbic acid and pyrroloquinoline quinone by weight, and mix them evenly at a low speed in a dry mixer; S2: Add pharmaceutically acceptable excipients to prepare an oral preparation according to the selected dosage form.

9. Use of the pharmaceutical composition according to claim 1 in the preparation of a medicament for treating alcohol addiction and / or alcohol dependence.

10. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for intervening in alcohol relapse or a drug for reducing alcohol intake.

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