KR20200050436A - Composition for preventing or treating methamphetamine addiction comprising alpha-pinene - Google Patents

Abstract

The present invention provides a composition for preventing or treating methamphetamine addiction containing α-pinene as an active component. The composition contains α-pinene, a natural material having proven stability, as the active component, and the α-pinene recovers a dopamine system damaged by methamphetamine and significantly inhibits damage or death of neurons caused by inflammation or oxidative stress promoted by methamphetamine, thereby being able to stably and effectively prevent, alleviate or treat methamphetamine addiction.

Description

Composition for preventing or treating methamphetamine poisoning containing alphapinene as an active ingredient {COMPOSITION FOR PREVENTING OR TREATING METHAMPHETAMINE ADDICTION COMPRISING ALPHA-PINENE}

The present invention relates to a composition for preventing or treating methamphetamine poisoning containing alpha-pinene as an active ingredient.

Recently, in Korea, people who are under stress due to rapid industrial development and complex social relations are increasing, and drugs are often used as a means to solve them. These drugs have psychological dependence, causing repeated drug abuse.

Methamphetamine (methamphetamine) is one of the most commonly used drugs in Korea, called methamphetamine, ice, and crystal. Methamphetamine is a drug that is derived from amphetamine and strongly induces excitation of the central nervous system.It is a trend of increasing abuse of methamphetamine worldwide because it can be easily made in small-scale facilities using relatively inexpensive OTC.

Methamphetamine blocks dopamine reuptake after binding to the dopamine transporter of systemic cells, so that dopamine binds to the dopamine receptors of the posterior neurons to continuously transmit excitation signals. It has been reported that this drug addiction is related to the reward circuit, and the neurotransmitter of the related pathway is projected from the ventral cranial region (VTA) to the lateral nucleus (NAc) back into the medial prefrontal cortex (mesolimbic dopamine). system).

According to statistical data reported by the Supreme Prosecutors' Office in September 2017, it was found that drugs from various social classes, from students to office workers and housewives, were abused, among which 30-40s accounted for 53.5% of drug abuse. In addition, methamphetamine seized from January to September 2017 was 18,422g, an increase of 23.3% compared to 2016, and is increasing every year.

Repeated administration of methamphetamine leads to changes in the dopamine system, mental disorders such as depression and anxiety, personality disorder, decreased memory, and hypersensitivity, and methamphetamine abusers have become a number of problems, such as mild or more depressed.

Korean Patent Publication No. 10-2006-0054904 (released on May 23, 2006)

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating methamphetamine poisoning containing alpha-pinene.

In addition, the present invention provides a health functional food composition for preventing or improving methamphetamine poisoning containing alphapinene (α-pinene).

The pharmaceutical composition for preventing or treating methamphetamine poisoning according to the present invention may contain alpha-pinene as an active ingredient.

The health functional food composition for preventing or improving methamphetamine poisoning according to the present invention may contain alpha-pinene as an active ingredient.

The present invention relates to a pharmaceutical composition and a health functional food composition for preventing or treating methamphetamine poisoning containing alphapinene, which is a natural product having proven stability, as an active ingredient, wherein the alphapinene restores the dopamine system damaged by methamphetamine, and methamphetamine. By significantly inhibiting the damage or death of nerve cells caused by inflammatory or oxidative stress promoted by, methamphetamine poisoning can be safely and effectively prevented, improved or treated.

1 is a graph showing the effect of alphapinene on conditional place preference (CPP) due to methamphetamine administration according to an experimental example of the present invention.
Figure 2 is a result of examining the change of dopamine-related protein by methamphetamine according to an experimental example of the present invention using Western blot analysis, the effect of alphapinene on the reduction of expression of dopamine synthase and D2 receptor by methamphetamine Shows.
3 is a result of examining the cell death, anti-cell death and inflammation-related protein expression changes by methamphetamine according to an experimental example of the present invention using Western blot analysis, neuronal cell death and microglia activation by methamphetamine. It shows the effect of alphapinene on.
Figure 4 is a result of examining the oxidative stress, transcription factor activation and antioxidant protein expression changes by methamphetamine according to an experimental example of the present invention using Western blot analysis.

Hereinafter, the present invention will be described in detail.

The present inventors selected alpha-pinene, an aromatic material derived from pine, which is proven to be stable through natural studies to develop safe and effective methamphetamine poisoning prevention or treatment drugs, and the alphapinene is caused by drug addiction. The present invention was completed by confirming that the place preference, the dopamine synthetase and receptor expression decreased, neuronal cell death and microglia activation, and oxidative damage were significantly inhibited.

The present invention provides a pharmaceutical composition for the prevention or treatment of methamphetamine poisoning containing alpha-pinene as an active ingredient.

As used herein, "alpha-pinene" is a terpene-based organic compound, (1S, 5S)-or (-)-α-pinene, (1R, 5R)-or (+)-α Two isomers of -isomer are well known. Alphapinene is found in the refining of conifers such as pine (Pinus densiflora) and rosemary (Rosmarinus officinalis), and phytoncide, a aromatic substance derived from pine, is the most representative component. In addition, it can be found in Artemisia vulgaris, Artemisia vulgaris, Chrysanthemum coronarium L. or Pepper nigrum, and Eucalyptus oils, Camphor, Camphor, Bupleurum fruiticescens or Opuntia humifusa It is contained in essential oils such as anti-cancer, anti-dementia, antioxidant, and anti-inflammatory.

The alphapinene may be extracted from an organism containing it, such as the natural source described above, chemically synthesized using a known manufacturing method, or commercially available one.

The alphapinene may be used in the form of a pharmaceutically or food acceptable salt within the range having the same efficacy.

In the present specification, "pharmaceutical or food acceptable" means that there is no toxicity to cells or humans exposed to the composition.

The salt may be used as either a pharmaceutically or food-acceptable basic salt or an acid salt. The basic salt can be used in either organic or inorganic base salts, sodium salt, potassium salt, calcium salt, lithium salt, magnesium salt, cesium salt, aluminum salt, ammonium salt, triethyl It can be selected from the group consisting of an aluminum salt and a pyridinium salt.

As the acid salt, an acid addition salt formed by free acid is useful. Inorganic and organic acids can be used as the free acid, hydrochloric acid, bromic acid, sulfuric acid, sulfurous acid, phosphoric acid, double phosphoric acid, nitric acid, etc. can be used as the inorganic acid. Citric acid, acetic acid, maleic acid, malic acid, and fumaric acid can be used as the organic acid. , Gluconic acid, methanesulfonic acid, benzenesulfonic acid, camphorsulfonic acid, oxalic acid, malonic acid, glutaric acid, acetic acid, glycolic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, Glutamic acid, citric acid, aspartic acid, stearic acid, and the like can be used, but are not limited thereto, and salts formed by using various inorganic and organic acids commonly used in the art may be included.

In addition, the alpha-pinene may include all salts, hydrates, solvates, derivatives, etc., which can be prepared by a conventional method, as well as pharmaceutically or food-acceptable salts. The addition salt can be prepared by a conventional method, after dissolving the alphapinene in a water-miscible organic solvent, such as acetone, methanol, ethanol, or acetonitrile, and adding an excess organic base or adding a base aqueous solution of an inorganic base It can be prepared by precipitation or crystallization. Or it can be prepared by evaporating the solvent or excess base from this mixture and then drying it to obtain additional salts or suction filtration of the precipitated salts.

As used herein, "methamphetamine" is a derivative of amphetamine, a synthetic chemical that causes arousal as a very powerful central nerve stimulant, so-called "hiropon (phylophone)". When administered, drowsiness and fatigue disappear, physical activity increases, and there is a risk of misuse due to feeling pleasure or happiness. It is classified as a psychotropic drug because it rapidly develops resistance, causes severe dependence upon continuous administration, and causes withdrawal symptoms when discontinued. It is not used as a drug in Korea and is a substance that is strongly regulated by law.

In the present specification, "prevention" refers to all actions to suppress the occurrence of methamphetamine poisoning, or a disease caused by the administration of the pharmaceutical composition or dietary supplement composition according to the present invention, or to thereby delay the onset. It also includes treatment of subjects with remission to the disease to prevent or prevent recurrence.

In the present specification, "treatment" means any act of improving or beneficially modifying the symptoms, such as alleviating, reducing, or abolishing methamphetamine poisoning or a disease caused by administration of the pharmaceutical composition according to the present invention.

In the present specification, "pharmaceutical composition" means a composition that is administered for a specific purpose, and is administered for the purpose of the present invention to prevent or treat methamphetamine poisoning, or diseases caused thereby.

In the pharmaceutical composition according to the present invention, the alphapinene can restore or improve the dopaminergic neurotransmission system (dopaminergic neural circuit) damaged by the methamphetamine. More specifically, the alphapinene may increase the expression of the dopamine synthase tyrosine hydroxyase (TH) and the dopamine binding D2 receptor reduced by the methamphetamine.

Methamphetamine increases the release of dopamine from the brain, and dopamine released in large amounts from the central nervous system raises mood and makes you feel pleasure. However, if an excessively large amount of dopamine is released into the synapse by the methamphetamine, a problem may arise in which the dopamine neural circuit is deformed.

The "dopamine" is a catecholamine-based organic compound, a neurotransmitter that transmits excitability of brain neurons, and a key material in addictive diseases. The dopamine is produced by synthesizing L-DOPA using tyrosine hydroxyase (TH) from tyrosine, and again removing a carboxyl group using DOPA decarboxylase. Dopamine released into synapses in various regions of the brain binds to dopamine receptors, thereby executing, motor control, motivation, arousal, reinforcement, and reward. ) And induces various functional changes. Typically, the dopamine D2 receptor (D2R) plays a central role in various neurophysiological functions.

According to one experimental example of the present invention, the expression of dopamine synthetase and D2 receptor was reduced by methamphetamine, but when alphapinene was administered, it was confirmed that the expression of the dopamine synthetase and dopamine receptor was significantly recovered. In order to maintain homeostasis due to excessive dopamine secretion by the methamphetamine, dopamine neural circuits such as dopamine synthetase and dopamine receptors are reduced or suppressed, but dopamine neural circuits modified or damaged by the alphapinene may be recovered. have. That is, the alphapinene can prevent or treat methamphetamine poisoning by restoring the dopamine nerve circuit damaged by the methamphetamine.

In the pharmaceutical composition according to the present invention, the alphapinene can protect the nerve cells by inhibiting the damage or death of the nerve cells by the methamphetamine.

Methamphetamine can damage or destroy brain neurons and degrade in the process of over-secreting dopamine.

According to an experimental example of the present invention, the control protein Bcl-2 (B-cell) that regulates cell death by inhibiting (anti-cell death) or inducing (pro-apoptotic) apoptosis by administration of methamphetamine. The expression of lymphoma 2) was decreased, the expression of apoptosis regulatory protein BAX, which is involved in cell activity, was increased, and the expression of inflammatory factor Iba-1 (ionized calcium-binding adapter molecule 1) was increased. However, when alphapinene was administered, it was confirmed that the expression of Bcl-2 increased again and the expression of BAX and Iba-1 decreased.

That is, the alpha-pinene may protect or prevent methamphetamine poisoning by protecting nerve cells by inhibiting the damage or death of the neurons by methamphetamine, and the inflammatory response.

In addition, according to another experimental example of the present invention, the expression of 4-hydroxylnonenal (4-HNE), an oxidative lipid damage and lipid peroxidation index, was increased by administration of methamphetamine, and the expression of antioxidant enzymes. Expression of transcription factors involved in p-Nrf2 (phosphorylated-Nrf2), antioxidant enzymes MnSOD and GCS decreased, but when alphapinene was administered, expression of 4-HNE decreased and p-Nrf2, MnSOD and GCS It was confirmed that the expression was increased.

That is, the alphapinene can prevent or treat methamphetamine poisoning by having a protective action against oxidative stress and damage to oxidative neurons such as increased lipid peroxidation by repeated administration of methamphetamine.

In the pharmaceutical composition according to the present invention, the alphapinene can suppress the conditional place preference (CPP) by the methamphetamine.

As used herein, “Conditioned Place Preference (CPP)” is a behavioral experiment in the form of a classic Pavlovian conditioning model, in which a specific space or situation is linked to addictive drug administration and another specific situation. , Or space is an experiment that analyzes which situation or animal prefers space at this time without drug administration in a subsequent test after training for conditioning by connecting to the administration of a control substance such as saline rather than an addictive drug. The degree of preference for a place or situation where an addictive drug has been administered can be said to indicate Pavlov's reinforcement effect on the drug, and is used to measure the craving for addictive drugs.

According to an experimental example of the present invention, the place preference of the group administered with methamphetamine was significantly increased compared to the control group administered with physiological saline, while the alphapinene administration group showed a significantly reduced place preference than the group administered with methamphetamine. , It can be seen that the place preference and compensation effect by methamphetamine is inhibited by alphapinene.

The pharmaceutical composition according to the present invention can be prepared according to conventional methods in the pharmaceutical field. The pharmaceutical composition may be formulated with a suitable pharmaceutically acceptable carrier according to the formulation, and, if necessary, may be prepared by further including excipients, diluents, dispersants, emulsifiers, buffers, stabilizers, binders, disintegrants, solvents, and the like. have. The appropriate carrier or the like does not inhibit the activity and properties of the alphapinene according to the present invention, or a pharmaceutically acceptable salt thereof, and may be selected differently according to the dosage form and formulation.

The pharmaceutical composition according to the present invention can be applied in any dosage form, and more specifically, it can be used by formulating into a parenteral dosage form of an oral dosage form, an external preparation, a suppository, and a sterile injectable solution according to a conventional method.

Among the oral dosage forms, the solid dosage form is in the form of tablets, pills, powders, granules, capsules, etc., at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, sorbitol, mannitol, cellulose, gelatin, etc. Can be prepared by mixing, and may include lubricants such as magnesium stearate and talc in addition to simple excipients. In addition, in the case of the capsul formulation, in addition to the above-mentioned substances, a liquid carrier such as fatty oil may be further included.

Among the oral dosage forms, the liquid dosage form is a suspension, an intravenous solution, an emulsion, a syrup, etc. In addition to water and liquid paraffin, which are commonly used diluents, various excipients, for example, wetting agents, sweeteners, flavoring agents, preservatives, etc. may be included. have.

The parenteral formulation may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used. Without being limited thereto, any suitable formulation known in the art may be used.

In addition, the pharmaceutical composition according to the present invention may further add calcium, vitamin D _3, and the like to improve the therapeutic efficacy.

In the pharmaceutical composition according to the present invention, the pharmaceutical composition may be administered in a pharmaceutically effective amount.

As used herein, “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and does not cause side effects.

The effective dose level of the pharmaceutical composition is intended for use, age of the patient, sex, weight and health status, type of disease, severity, drug activity, sensitivity to the drug, administration method, administration time, administration route and discharge rate, treatment The duration, combination, or factors including the drug used concurrently and other factors well known in the medical field can be determined differently. For example, although not constant, it is generally 0.001 to 100 mg / kg, preferably 0.01 to 10 mg / kg once to several times a day. The above dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition according to the present invention may be administered to any animal in which methamphetamine poisoning may occur, and the animal may include, for example, humans and primates, as well as livestock such as cattle, pigs, horses, and dogs. .

The pharmaceutical composition according to the present invention can be administered by a suitable route of administration according to the form of the formulation, and can be administered through various routes, oral or parenteral, as long as it can reach the target tissue. The method of administration is not particularly limited, and may be administered by conventional methods such as, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrabronchial inhalation, intrauterine epidural or intracere-broventricular injection. .

The pharmaceutical composition according to the present invention may be used alone for the prevention or treatment of methamphetamine poisoning, and may be used in combination with surgery or other drug treatment.

In addition, the present invention provides a health functional food composition for preventing or improving methamphetamine poisoning containing alpha-pinene as an active ingredient.

In the present specification, "improvement" means any act of improving or beneficially improving the symptoms, such as alleviating, reducing, or extinguishing methamphetamine poisoning or a disease caused by ingestion of the health functional food composition according to the present invention. do.

In the present specification, the term "health functional food" includes food manufactured and processed using ingredients or ingredients having useful functions for the human body according to Act No. 6727 of the Health Functional Food, and in addition to nutrition, the purpose of the present invention It means food with high medical and medical effects processed to effectively exhibit bio-regulatory functions such as prevention of methamphetamine poisoning, bio-defense, immunity, and recovery.

In the dietary supplement composition according to the present invention, the alphapinene can restore or improve the dopaminergic neurotransmitter system damaged by the methamphetamine. More specifically, the alphapinene may increase the expression of the dopamine synthase tyrosine hydroxyase (TH) and the dopamine binding D2 receptor reduced by the methamphetamine.

In addition, the alphapinene can protect the nerve cells by inhibiting the damage or death of the nerve cells by the methamphetamine. The alphapinene suppresses damage or death of nerve cells caused by methamphetamine, suppresses inflammatory reactions, and has a protective action against oxidative nerve cell damage such as oxidative stress and increased lipid peroxidation, thereby preventing or improving methamphetamine poisoning. It can be utilized as a functional food composition.

In the dietary supplement composition according to the present invention, the dietary supplement may be prepared as a powder, granule, tablet, capsule, syrup or beverage, and there is no limitation in the form that the dietary supplement can take, and is conventional It can include all foods of meaning. For example, drinks and various drinks, fruits and processed foods thereof (canned fruit, jam, etc.), fish, meat and processed foods (ham, bacon, etc.), breads and noodles, cookies and snacks, dairy products (butter, cheese, etc.) ) And the like, and may include all functional foods in a conventional sense. It may also include food used as feed for animals.

The nutraceutical composition according to the present invention may be prepared by further comprising a food additive that is commonly used in the art, a food additive (food additive), and other suitable ingredients. Compliance with food additives can be determined according to the standards and standards for the product in accordance with the General Rules and General Test Methods for Food Additives approved by the Food and Drug Administration, unless otherwise specified.

The other auxiliary ingredients include, for example, flavoring agents, natural carbohydrates, sweeteners, vitamins, electrolytes, colorants, pectic acids, alginic acids, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid, etc. It may further contain. In particular, as the natural carbohydrate, monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol, and erythritol may be used. , As a sweetener, natural sweeteners such as taumatin and stevia extract, synthetic sweeteners such as saccharin and aspartame can be used.

The effective dose of the alphapinene contained in the dietary supplement according to the present invention, or a pharmaceutically acceptable salt thereof, can be appropriately adjusted according to the purpose of use such as prevention or improvement of methamphetamine poisoning.

The composition has the advantage that there is no side effect, etc. that may occur when taking a long-term use of a general drug as a raw material, and is excellent in portability, and may be taken as a supplement for preventing or improving methamphetamine poisoning.

Hereinafter, examples will be described in detail to help understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

<Preparation for Experiment>

Male C57BL6 / J mice, around 20-23 g of 6 weeks of age, were supplied from Hyochang Science (Daegu) and adapted to a one-week breeding room environment. The laboratory animals were kept in the laboratory animal room at Keimyung University School of Medicine at 22 ± 1 ℃ and the humidity was 50 ± 5%. Was made possible. The experimental animals were divided into 4 groups of 5 groups, each of the control group and the test group. During the experiment, 1 mg / kg of methamphetamine purchased from the Ministry of Food and Drug Safety and 3 mg / kg or 10 mg / of alphapinene purchased from Sigma-Aldrich, USA It was dissolved in physiological saline in the amount of kg and administered into the abdominal cavity of mice.

<Statistics processing method>

SPSS program (22.0, Korean version) one-way ANOVA was used to check the statistical significance, and post-test with LSD (least significance difference) showed that the p-value was less than 0.05.

<Experimental Example 1> Inhibition of drug dependency of alphapinene on methamphetamine-induced drug addiction

In order to confirm the inhibitory effects of drug addiction and alphapinene by methamphetamine, it was measured using a conditioned place preference (CPP) method, which is a representative experimental method.

1. Experimental device

The conditional place preference test device consists of two boxes: a white box and a black box with a width (15cm) × a height (15cm) × a height (15cm), and each room has a horizontal network and a cross network so that mice can remember it The guillotine door was placed and opened and closed according to the experiment.

2. Experimental method

On the first and second days, the mice were opened with guillotine doors, allowing the white and black boxes to move freely for 20 minutes, allowing them to explore and adapt to the boxes, and measure the time to stay in each box on the third day to measure the mouse's box Preference was confirmed (Pre-CPP).

The mice were then intraperitoneally administered 1 mg / kg of methamphetamine on the 4th, 6th, 8th, and 10th days, and placed in a white or black box for 45 minutes to allow the mice to remember the drug administration and the place, and on the 5th, 7th, 9th and 11th days. After the saline was administered, it was allowed to remain in the opposite box for 45 minutes. Alphapinene was administered 3mg / kg or 10mg / kg intraperitoneally 30 minutes before administration of methamphetamine (conditioning). On the 12th, the guillotine door was opened again to allow the white box and the black box to move freely for 20 minutes, and the time spent in each box was recorded for 20 minutes (Post-CPP).

3. Experimental results

1 is a graph showing the effect of alphapinene on conditional place preference (CPP) due to methamphetamine administration according to an experimental example of the present invention.

Referring to FIG. 1, the place preference of the control group administered with physiological saline was 74.678 seconds, and the group administered with methamphetamine (1 mg / kg) was 285.82 seconds, confirming that the place preference was significantly increased. On the other hand, the 3 mg / kg alphapinene administration group was found to be slightly lower than the methamphetamine administration group at 168.79 seconds, and the 10 mg / kg alphapinene administration group was 104.14 seconds, which significantly decreased the place preference compared to the methamphetamine group. The above results show that place preference and compensation effects by methamphetamine are inhibited by alphapinene.

<Experimental Example 2> Confirmation of molecular marker expression according to methamphetamine-induced drug poisoning of alphapinene

1. Experimental method

Western blot was performed to measure the change in expression of molecular indicators related to drug addiction. After the behavioral experiment, the brain's brain is separated from the striatum and cerebral cortex, and ice-cold RIPA buffer; 150 mM NaCl, 0.5% Triton X-100, 50 mM The tissue was homogenized with Tris-HCl, pH 7.4, 25 mM NaF, 20 mM EGTA, 1 mM DTT, 1 mM Na3VO4, protease inhibitor cocktail).

After lysis at 4 ° C for 30 minutes, the supernatant was taken by centrifugation at 14,000 g for 15 minutes, and then the protein content was quantified with a BCA reagent, and 35 μg was electrophoresed on 12% SDS-PAGE. The developed gel was transferred to a PVDF membrane (Roche, USA) and then blocked with 5% non-fat dry milk-PBST (PBSST, PBS, 0.1% Tween-20). Then, attach the primary antibody to 3% skim milk-PBS (non-fat dry milk-PBS) overnight, wash 3 times with PBST solution, add secondary antibody to 3% skim milk-PBS for 1 hour, and then attach PBST. The solution was washed 3 times for 10 minutes again.

Thereafter, the reaction was performed for 1 minute using an enhanced chemoluminescence (ECL) solution (Amersham Pharmacia Biotech., IL, USA), and then photographed using a Chemiluminescent Immunoblotting Imaging (Fujifilm, Japan) equipment.

2. Experimental results

Figure 2 is a result of examining the change of dopamine-related protein by methamphetamine according to an experimental example of the present invention using Western blot analysis, the effect of alphapinene on the decrease in the expression of dopamine synthase and D2 receptor by methamphetamine Shows.

Referring to FIG. 2, it was shown in FIG. 2A that the expression of tyrosine hydroxylase (TH), an enzyme involved in dopamine production, was significantly reduced in the striatal region of the methamphetamine (1 mg / kg) administration group compared to the control group. It was found that expression was significantly restored by administration of alphapinene.

In FIG. 2B, the expression of the D2 receptor, one of the dopamine-binding major receptors, was found to be decreased in the methamphetamine administration group compared to the control group, which was significantly increased by alphapinene administration. The relative expression level of the protein was quantified to show statistical significance in the graph on the right.

Figure 3 is a result of examining the cell death, anti-cell death and inflammation-related protein expression changes by methamphetamine according to an experimental example of the present invention using Western blot analysis, neuronal cell death and microglia activation by methamphetamine It shows the effect of alphapinene on.

Referring to FIG. 3, it was confirmed in FIG. 3A that the expression of the anti-apoptotic indicator protein Bcl-2 was decreased in the methamphetamine administration group compared to the control group, and the expression of the apoptosis-mediated protein indicator Bax was increased. On the other hand, it was confirmed that the expression of reduced Bcl-2 in the alphapinene administration group was increased compared to the methamphetamine administration group, and the expression of Bax was decreased.

In FIG. 3B, the expression of Iba-1, a microglia activation protein, which is a representative inflammatory-related indicator, was increased in the methamphetamine administration group compared to the control group, and it was confirmed that this was significantly reduced in the alphapinene administration group, especially 10 mg / kg.

Through the above results, neuronal cell death is induced due to decreased Bcl-2 expression and increased Bax and Iba-1 protein expression upon repeated administration of methamphetamine, but increased Bcl-2 protein expression after administration of alphapinene, Bax and Iba- 1 By reducing protein expression, it can be determined that alphapinene suppresses neuronal cell death and inflammatory response and has a protective action against neuronal damage caused by drug addiction. The relative expression level of the protein was quantified to show statistical significance in the graph on the right.

Figure 4 is a result of examining the oxidative stress, transcription factor activation and antioxidant protein expression changes by methamphetamine according to an experimental example of the present invention using Western blot analysis.

Referring to FIG. 4, the expression of 4-hydroxylnonenal (4-HNE), which is a representative oxidative lipid damage and lipid peroxidation index in FIG. 4A, was increased in the methamphetamine administration group compared to the control group, which is alphapinene 3mg It was found to be significantly reduced in the / kg and 10mg / kg administration groups.

In FIG. 4B, expression of p-Nrf2, a transcription factor involved in the expression of antioxidant enzymes, was decreased in the methamphetamine administration group compared to the control group, but was significantly increased in the alphapinene administration group, and the representative antioxidant enzymes MnSOD and GCS in FIG. 4C. The expression of the protein was decreased in the methamphetamine administration group compared to the control group, but was significantly increased in the alphapinene administration group.

Through the above results, oxidative stress was induced by repeated administration of methamphetamine, leading to oxidative neuronal damage such as increased lipid peroxidation, but activation of Nrf2 after alphapinene administration and then increased expression of antioxidant enzymes MnSOD and GCS protein. It can be determined that alphapinene has a protective action against neuronal damage caused by drug addiction. The relative expression level of the protein was quantified to show statistical significance in the graph on the right.

The formulation example of the composition containing alphapinene according to the present invention is described below, but the present invention is intended to be described in detail rather than to limit it.

<Production Example 1> Preparation of powder

Alpha-pinene 300 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight fabric to prepare a powder.

<Formulation Example 2> Preparation of tablets

Alpha-pinene 300 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to a conventional tablet manufacturing method.

<Formulation Example 3> Preparation of capsules

Alpha-pinene 300 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into a gelatin capsule to prepare a capsule.

<Formulation Example 4> Preparation of injection

Alpha-pinene 300 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na2HPO412H2O 26 mg

It is prepared with the above-mentioned ingredient content per ampoule (2 ml) according to the preparation method of a conventional injection.

<Formulation Example 5> Preparation of liquid formulation

Alpha-pinene 300 mg

Isomerized sugar 10 g

5 g mannitol

Purified water

Each component was added to the purified water to dissolve it according to the method of preparing a conventional liquid formulation, lemon lemon was added in an appropriate amount, and then the above components were mixed and purified water was added to adjust the total to 100 ml. Sterilization to prepare a liquid.

<Formulation Example 6> Preparation of healthy food

Alpha-pinene 1000 mg

Vitamin mixture

Vitamin A Acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ folic acid

Calcium Pantothenate 0.5 mg

Suitable amount of mineral mixture

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium phosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the vitamin and mineral mixture is a composition suitable for a relatively healthy food in a preferred embodiment, the composition ratio may be arbitrarily modified. , Granules can be prepared and used in the preparation of healthy food compositions according to conventional methods.

<Formulation Example 7> Preparation of healthy beverages

Alpha-pinene 300 mg

Vitamin C 15 g

Vitamin E (powder) 100 g

Iron lactate 19.75 g

Zinc oxide 3.5 g

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3 g

Water metering

After mixing the above ingredients according to a conventional health drink manufacturing method, and stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered, obtained in a sterilized 2L container, sealed and sterilized, then stored in a refrigerator and then stored in the present invention. It is used for the preparation of healthy beverage compositions.

Although the above composition ratio is a mixture of components suitable for a preference drink in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

Since the specific parts of the present invention have been described in detail above, for those skilled in the art, it is obvious that this specific technique is only a preferred embodiment, and the scope of the present invention is not limited thereby. Do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (6)

Pharmaceutical composition for preventing or treating methamphetamine poisoning containing alpha-pinene as an active ingredient. According to claim 1,
The alphapinene,
A pharmaceutical composition for the prevention or treatment of methamphetamine poisoning, characterized by increasing the expression of tyrosine hydroxyase (TH) and dopamine-binding D2 receptors reduced by the methamphetamine. According to claim 1,
The alphapinene,
A pharmaceutical composition for preventing or treating methamphetamine poisoning, characterized by protecting nerve cells by inhibiting damage or death of nerve cells by the methamphetamine. According to claim 1,
The alphapinene,
A pharmaceutical composition for preventing or treating methamphetamine poisoning, characterized by inhibiting conditional place preference (CPP) by the methamphetamine. A health functional food composition for preventing or improving methamphetamine poisoning containing alpha-pinene as an active ingredient. The method of claim 5,
The alphapinene,
A health functional food composition for preventing or improving methamphetamine poisoning, characterized by protecting nerve cells by inhibiting damage or death of nerve cells caused by the methamphetamine.

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