KR20220104542A - COMPOSITION FOR pain relief AND PREVENTING OR ALLEVIATING STRESS-INVOLVED DISEASE COMPRISING Extract of CANNABIS SATIVA L. - Google Patents

Abstract

The present invention relates to a composition for pain relief and stress-related disease prevention and alleviation comprising an extract of Cannabis sativa L. as an active ingredient. The composition for pain relief and stress-related disease prevention and alleviation inhibits cyclooxygenase (COX)-2 enzyme and simultaneously prevents dopamine and serotonin from being inhibited by using an extract of Cannabis sativa L.

Description

Composition for pain relief and prevention and improvement of stress-related diseases comprising hemp extract as an active ingredient

The present invention relates to a composition for relieving pain and preventing and improving stress-related diseases comprising a hemp extract as an active ingredient. More particularly, it relates to a composition for relieving pain and preventing and improving stress-related diseases, which inhibits cyclooxygenase (COX)-2 enzymes and at the same time prevents dopamine and serotonin from being inhibited using a hemp extract.

As the present society rapidly develops and diversifies, modern people are demanding various roles. Accordingly, the number of people complaining of anxiety disorders and mental disorders due to various stresses is increasing. Considering the recent trend of increasing mental illness among adolescents due to excessive academic enthusiasm or various kinds of stress, it can be seen that various kinds of stress and related mental disorders are gradually increasing.

Here, looking briefly at the mechanism of changes in stress-related neurotransmitters and hormones, the hypothalamus in the body controls the secretion of neurotransmitters and hormones in response to external stimuli and stress, and signals from the hypothalamus to the central nervous system ), the release of neurotransmitters such as dopamine, noradrenaline, and serotonin from each nerve terminal is regulated, thereby regulating physiological activities such as emotional state, heart rate, blood pressure, and blood flow of skeletal muscle. will become

In addition, hormones are secreted into the blood through the hypothalamic-pituitary-adrenal (HPA) axis system, which is a circulatory system of hormones from the hypothalamus to the pituitary-adrenal gland. If a human is subjected to an external stimulus or stress, the hypothalamus in the body secretes corticotropin-releasing factor (CRF, adrenocorticotropin releasing factor). Then, when the secreted CRF binds to a receptor that specifically binds to the CRF of the pituitary gland, adrenocorticotropic hormone (ACTH, adrenocorticotropic hormone) is secreted.

When the secreted ACTH arrives at the adrenal gland through the blood and lymph nodes, steroid hormones such as corticosterone and catecholamine-type hormones are released from the cortex into the blood, and are involved in the regulation of heart rate, blood pressure and energy metabolism. Moreover, secretion regulation of these factors is being achieved through negative feedback regulation.

Serotonin is a neurometabolite found in cerebrospinal fluid that circulates in the brain and functions as a neurotransmitter. The serotonin is closely related to emotional expression, and when this substance is insufficient, emotions are unstable, so anxiety, worry, and impulsive tendencies appear. The serotonin deficiency is closely related to depression, and there are many drugs currently used as antidepressants that prevent reabsorption of serotonin and stay in the brain for a longer period of time.

On the other hand, depression as used in the present invention is defined as not only the dictionary meaning of depression, but also the meaning including anxiety or anxiety disorder.

Anxiety is a mental disease that shows irritability, tension, anxiety, etc. that is persistent and does not usually recover over 6 months or more. This is usually caused by structural and functional abnormalities of the brain and continuous stress, and in particular in children and adolescents, when attention deficit hyperactivity disorder (ADHD) is severe, it can lead to anxiety and depression. Anxiety is broadly classified into four types: generalized anxiety disorder, obsessive-compulsive disorder, panic disorder, and post-traumatic stress disorder. and autonomic hypersensitivity symptoms appear as the most characteristic features.

Currently, the treatment of anxiety in clinical practice is being treated with medication and long-term psychotherapy. In the case of drug treatment, benzodiazepine anti-anxiety drugs such as diazepam, lorazepam, clonazepam, and alprazolam are mainly used. It is being used as a drug. In addition, drugs of the imidazopyridine class, such as zolpidem, are also used to treat short-term insomnia caused by anxiety.

However, most of the drugs used for anxiety disorders and psychiatric disorders are classified as psychotropic drugs. In particular, benzodiazepine drugs act directly on gamma-aminobutyric acid (GABA) receptors in the central nervous system to suppress the central nervous system. Excessive sedation may appear, and GABA inhibitory action may be enhanced, and central depressant actions such as depression, muscle relaxation, and hypnosis may be enhanced. In addition, drugs used for anxiety disorders and psychiatric disorders pose risks such as fear of drug abuse by making them psychologically and physically dependent.

Therefore, there is a need to develop a health functional food that has an effect on mental illness, does not cause excessive sedation and drowsiness, and does not cause dependence or a health functional food that can prevent a stress stimulus from developing into a mental disease.

In addition, these stressful stimuli may indicate a headache with a feeling of tightness, heaviness, or throbbing of the head. Usually, it is a feeling of pain on both sides of the head, and drug therapy is mainly used for treatment.

KR 10-2010-0106062 A

It is an object of the present invention to provide a composition for relieving pain and preventing and improving stress-related diseases comprising a hemp extract as an active ingredient.

Another object of the present invention is to provide a composition for alleviating pain and preventing stress-related diseases and amelioration at the same time as inhibiting the cyclooxygenase (COX)-2 enzyme of the hemp extract while inhibiting dopamine and serotonin.

Another object of the present invention is to provide a composition for alleviating pain and preventing and ameliorating stress-related diseases and having excellent effects of pain relief and prevention and improvement of stress-related diseases, and at the same time, no side effects occur during administration or administration.

In order to achieve the above object, the composition for alleviating pain and preventing and improving stress-related diseases according to an embodiment of the present invention includes cannabis sativa L. extract as an active ingredient.

The hemp extract contains cannabinoids and terpenes.

The stress-related disease will be any one selected from the group consisting of stress, depression and insomnia.

The composition inhibits the cyclooxygenase (COX)-2 enzyme to exhibit a pain relief effect.

The composition is to prevent and improve stress-related diseases by preventing dopamine and serotonin from being inhibited.

The extract includes extraction using an extraction solvent selected from the group consisting of water, C ₁ to C ₆ lower alcohols, and mixtures thereof.

Functional food according to another embodiment of the present invention is prepared by using a composition for alleviating pain and preventing and improving stress-related diseases comprising hemp extract as an active ingredient.

The external preparation for skin according to another embodiment of the present invention is prepared using a composition for alleviating pain and preventing and improving stress-related diseases comprising a hemp extract as an active ingredient.

The pharmaceutical according to another embodiment of the present invention is prepared by using a composition for alleviating pain and preventing and improving stress-related diseases comprising a hemp extract as an active ingredient.

Hereinafter, the present invention will be described in more detail.

As used herein, the term 'extract' has the meaning commonly used as a crude extract in the art as described above, but in a broad sense also includes a fraction obtained by additionally fractionating the extract. That is, the extract includes not only those obtained using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity), etc. The fraction obtained through the purification method is also included in the extract of the present invention.

In order to achieve the above object, the composition for alleviating pain and preventing and improving stress-related diseases according to an embodiment of the present invention includes cannabis sativa L. extract as an active ingredient.

Cannabis sativa L. is an annual plant of the genus Cannabaceae, and its subspecies include C. sativa subsp. sativa, C. sativa subsp. indica, C. sativa subsp. ruderalis are classified into three. It is known that it was widely cultivated throughout tropical and temperate regions, including Central Asia, for about 12,000 years in the period when legal regulations were not applied in the past, and it was used for food in China around 6,000 BC. There is a record that fiber was obtained from hemp around 4,000 BC and used, and in Chinese herbal books, there is a document that hemp was first used for medical purposes in 2,727 BC. However, in the 20th century, cannabis cultivation and handling are strictly regulated not only in Korea but also in most countries due to side effects such as hallucinations. However, overseas, the importance of the excellent pharmacological action of cannabis is gradually recognized, and the movement to allow the use of cannabis for therapeutic purposes is increasing. In Korea, there is a lack of research on cannabis due to strict regulations on cannabis.

Hemp has been used in a variety of ways in the past depending on the part, but the hemp leaf, cannabis or hemp leaf, is said to have the effect of killing roundworms. It is also used for asthma and old cough, roundworm, analgesic, anesthetic, and diuretic. The root of hemp was said to cure dyspareunia and the absence of placenta with hemp root, to relieve eohyeol and to release seokrim. Hemp, which is the skin of hemp, is said to treat bruises and open sores, and hemp, the flower of hemp, is said to be used for paralysis and itching. Ears of hemp are called powder, and are used for difficult births, constipation, gout, jingwang, and insomnia.

The hemp extract contains cannabinoids and terpenes.

To date, about 400 compounds have been discovered in cannabis, most of which are cannabinoids, terpenes, and phenolic compounds. Among them, cannabinoids are known as representative active ingredients of cannabis, and about 90 types of cannabinoids have been identified so far, and many of the ingredients found only in cannabis are known. Cannabinol (CBN) was isolated from hemp in 1899, but it was later known that it was not a single compound, and cannabidiol (CBD) and tetrahydrocannabinol (tetrahydrocannabinol, THC) was isolated, and research on the components of hemp became more active.

Efforts to develop drugs using specific components of cannabis are continuing, and THC and CBD, the main compounds of medical hemp, are receiving the most attention for therapeutic purposes. In the case of CBD, there are studies that show that it has no psychoactive effect, and that it has obtained effective results in relieving pain and controlling epileptic seizures.

In addition, more than 100 compounds of the terpene family, which play a role in giving the aroma and taste of hemp, have also been identified in hemp, and in the form of various monoterpenoids and sesquiterpenoids. exist. Terpenes have been found to be related to various pharmacological actions such as anti-inflammatory action, but studies on terpene compounds extracted from cannabis are still insufficient compared to THC.

In addition, terpenes are known to be more effective when combined with cannabinoids such as CBD and THC, improving absorption of cannabinoids, overcoming bacterial defense mechanisms, and reducing side effects. can be minimized

The stress-related disease will be any one selected from the group consisting of stress, depression and insomnia.

Stress refers to the feelings of anxiety and threat that humans feel when they are in psychologically or physically difficult situations.

Depression is a disease in which 'negative emotions' appear due to changes in the brain's ability to control emotions, and it is a disease that affects more than 100 million people around the world. The essence of depression is a physiological and anatomical problem, and negative emotions are the result of such changes in the structure of the human body. Feeling temporarily depressed is called 'depression'. The causes of depression include interpersonal stress and economic problems.

Insomnia (sleep disorder) refers to a sleep disorder in which it is difficult to fall asleep and a sleep maintenance disorder in which a person falls asleep but wakes frequently or wakes up too early. If you do not get enough sleep at night, you will become sleep deprived, causing drowsiness, fatigue, and loss of motivation during the day, disrupting your daily life and lowering your quality of life. Insomnia occurs in people who have irregular sleeping times or habits, and the symptoms worsen as they experience environmental changes and psychological stress. Even if you worry too much about insomnia itself, the nervous system is tense and insomnia can persist and worsen. There are various causes of insomnia, but common causes of temporary insomnia are jet lag due to travel, new job (work), moving, hospitalization. It will change your regular life rhythm.

If the stress-related disease is chronic, symptoms such as pain, arthritis, headache, and breathing difficulties may be accompanied by insomnia.

Therefore, the present invention can prevent and improve stress-related diseases as described above while exhibiting a pain relief effect using cannabis extract.

The composition is to prevent and improve stress-related diseases by preventing dopamine and serotonin from being inhibited.

Dopamine is a neurotransmitter that is a precursor to the synthesis of norepinephrine and epinephrine. It is one of the amino acids present in animals and plants, and plays a role in excitation transmission of brain nerve cells. In our brain, dopamine acts as a neurotransmitter. Dopaminergic neurons that secrete dopamine are located in the ventral tegmental area of the midbrain, the substantia nigra, and the arcuate nucleus of hypothalamus. In the ventral tegmental area, dopamine innervation is controlled by the cerebral cortex and limbic system, and these are called mesocortical and mesolimbic pathways, respectively. In the substantia nigra, dopamine innervates the striatum, which is called the nigrostriatal pathway. Dopaminergic neurons located in the hypothalamus synovial nucleus (TIDA neurons) dopamine innervation through the tuberoinfundibular pathway to the median elevation.

Dopamine molecules bind to dopamine receptors in neurons, and dopamine receptors bind to G-protein (GTP-binging-protein) to activate second messengers or activate or inhibit specific signaling pathways in cells. It is possible to control the degree to which is excited or inhibited.

Also, serotonin is one of the chemicals that functions as a neurotransmitter present in the hypothalamus of the brain and is closely related to emotional expression. When this substance is lacking, emotions are unstable, causing more anxiety and worry, impulsive tendencies, and a molecule that makes you feel happy. In addition to regulating mood, serotonin is involved in many functions related to appetite, sleep, and muscle contraction. It is also related to thinking function, which affects memory and learning, and is stored in platelets and is involved in hemostasis and blood clotting reactions. It is synthesized from L-tryptophan through a short pathway, and tryptophan hydroxylase and amino acid decarboxylase are involved in this reaction.

A lack of serotonin can lead to depression and anxiety. It also acts as an important regulator in appetite and food selection and is known to be most related to carbohydrate intake. When serotonin increases locally, appetite decreases, and when it decreases, the opposite phenomenon occurs.

Therefore, the composition for alleviating pain and preventing and improving stress-related diseases of the present invention can exhibit the effect of preventing and improving stress-related diseases by preventing dopamine and serotonin from being inhibited by including hemp extract as an active ingredient.

The composition inhibits the cyclooxygenase (COX)-2 enzyme to exhibit a pain relief effect.

Cyclooxygenase (COX)-2 is an enzyme that induces the synthesis of prostaglandin (PG), a pain-causing substance, and suppresses the expression of COX-2 because the synthesis of NO is promoted by other stimuli. Inhibiting or inhibiting activity is also an important marker in the treatment of pain.

Inflammatory response is induced by harmful stimuli or infection, and depending on the type of cell and stimulus, prostaglandin (PG), leukotriene (LT), etc. are produced, and cyclooxygenase-1 (COX-1) and cyclooxygenase- 2 (COX-2). COX-1 and COX-2 are isoforms with almost identical identity, and COX-1 exists in most cells and acts on normal biological functions involved in gastric mucosal protection, platelet coagulation, and renal blood flow, whereas COX -2 is induced and expressed in a short time by inflammatory stimuli, and is an enzyme present in macrophages and mononuclear cells that are amplified under inflammatory conditions and is known to be deeply related to inflammation, pain, fever, and anemia.

Therefore, the composition for alleviating pain and preventing and improving stress-related diseases of the present invention contains hemp extract as an active ingredient, and may exhibit a pain relief effect by inhibiting the cyclooxygenase (COX)-2 enzyme.

Preferably, the composition for alleviating pain and preventing and improving stress-related diseases may further include an annual extract, carob extract, and star flower extract.

The Erigeron annuus (L.) Pers is native to North America. It is a plant of the genus Chrysanthemum that grows in fields, fields, and roadsides in various places in Korea. Various physiological activities are known.

The carob {Ceratonia siliqua (L.) Taub.} is an evergreen broad-leaved shrub that grows to about 10 m in height and tolerates drought well. The leaves contain resin. Small, dull greenish-brown flowers bloom in autumn, with long green pods running. These pods turn chocolate when ripe and contain small, shiny hard beans. Carob gum, which softens the skin, is popular as a skin pack and is also used to treat diarrhea.

The star flower (Stellaria media) is a two-year-old plant of the family Seokjukdae and grows mainly on the roadside or on the banks of the field, and the height is about 10-20 cm, and contains abundant minerals such as protein, calcium, and iron. This star flower strengthens the stomach, purifies the blood, and produces milk well. It is also used to treat inflammations such as gum disease, tooth decay, appendicitis, enteritis, and intestinal ulcers.

When the natural extract is used in combination, the effect of relieving pain and preventing and improving stress-related diseases is increased due to the mixing action between each component.

In addition, by additionally including the annual extract, carob extract, and star flower extract, it neutralizes the unique taste and flavor of the hemp extract, thereby enabling it to be provided as a composition with excellent palatability.

Preferably, the composition of the present invention may include 20 to 40 parts by weight of the annual alpine extract, 20 to 40 parts by weight of the carob extract, and 20 to 40 parts by weight of the star flower extract based on 100 parts by weight of the hemp extract. When used as a complex extract within the above range, it exhibits excellent pain relief and prevention and improvement effects of stress-related diseases, while enabling the provision of a composition with excellent palatability.

The extract includes extraction using an extraction solvent selected from the group consisting of water, C ₁ to C ₆ lower alcohols, and mixtures thereof.

Specifically, pulverizing a natural product to prepare a hemp extract; leaching the pulverized product using an organic solvent; drying the sample after leaching; re-leaching the dried sample using an organic solvent; drying the sample after leaching; leaching with water; And including the step of leaching, it is possible to obtain a natural extract.

The natural extract extracted using the organic solvent may further include the step of performing fractionation using an organic solvent.

The extraction solvent may be used in an amount of 2 to 50 times, more specifically, 2 to 20 times, based on the weight of the sample. For extraction, the sample may be left for 1 to 72 hours for leaching in the extraction solvent, and more specifically, for 24 to 48 hours.

The extract may be prepared in a powder state by additional processes such as vacuum distillation and freeze drying or spray drying, and is obtained by an extraction method selected from the group consisting of solvent extraction method, ultrasonic extraction method, reflux extraction method, leaching method, fermentation method and foaming method. include that

In the ultrasonic extraction method, the reaction is performed at 30 to 50° C. for 0.5 to 2.5 hours, and the extraction solvent is water or 50 to 100% alcohol having 1 to 6 carbon atoms. Specifically, extraction is performed at 40 to 50° C. for 1 to 2.5 hours, and the extraction solvent is water or 70 to 80% alcohol having 1 to 6 carbon atoms.

The reflux extraction method is based on 100 mL of water, an alcohol having 1 to 6 carbon atoms, 10 to 30 g of a pulverized product of a natural product, a reflux time of 1 to 3 hours, and 50 to 100% alcohol or water having 1 to 6 carbon atoms. More specifically, based on 100 mL of alcohol having 1 to 6 carbon atoms or 100 mL of water, 10 to 20 g of a pulverized product of natural products, 1 to 2 hours reflux, and 70 to 90% alcohol or water having 1 to 4 carbon atoms.

The leaching method is carried out at 15 to 30° C. for 24 to 72 hours, and water or 50 to 100% alcohol having 1 to 6 carbon atoms is used as an extraction solvent. More specifically, the process is carried out at 20 to 25° C. for 30 to 54 hours, and the extraction solvent is water or 70 to 80% alcohol having 1 to 6 carbon atoms.

After extraction, the extract may be fractionated by sequentially applying a fresh fractionation solvent. The fractionation solvent used for fractionation is any one or more selected from the group consisting of water, hexane, butanol, ethyl acetic acid, ethyl acetate, methylene chloride and mixtures thereof, preferably ethyl acetate or methylene chloride.

Functional food according to another embodiment of the present invention is prepared by using a composition for alleviating pain and preventing and improving stress-related diseases comprising hemp extract as an active ingredient.

"Functional food" as defined herein means a food manufactured and processed using raw materials or ingredients useful for the human body according to Act No. 6727 of the Functional Food Act, and "functionality" refers to the structure and It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients for function or physiological effects.

The external preparation for skin according to another embodiment of the present invention is prepared using a composition for alleviating pain and preventing and improving stress-related diseases comprising a hemp extract as an active ingredient.

The pharmaceutical according to another embodiment of the present invention is prepared by using a composition for alleviating pain and preventing and improving stress-related diseases comprising a hemp extract as an active ingredient.

The dosage form for the pharmaceutical of the present invention may be in a desired form depending on the method of use, and in particular, it may be formulated by adopting a method known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. can Examples of specific formulations may be granules, powders, syrups, solutions, suspensions, ointments, needles, tablets, suppositories, injections, spirits, capsules, pills, soft or hard gelatin capsules, and the like.

The preferred dosage for the drug of the present invention varies depending on the condition and body weight, the degree of disease, the form of the drug, the route and duration of administration, but may be appropriately selected by those of ordinary skill in the art to which the present invention pertains. have.

According to the composition for pain relief and prevention and improvement of stress-related diseases comprising the hemp extract of the present invention as an active ingredient, the use of hemp extract inhibits cyclooxygenase (COX)-2 enzymes and simultaneously inhibits dopamine and serotonin It is possible to provide a composition for alleviating pain and preventing and improving stress-related diseases.

1 is an experimental result for pain relief of a composition according to an embodiment of the present invention.
2 is a result of a dopamine measurement experiment of the composition according to an embodiment of the present invention.
3 is a result of a serotonin measurement experiment of the composition according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those of ordinary skill in the art can easily carry out the present invention. However, the present invention may be embodied in several different forms and is not limited to the embodiments described herein.

[Preparation Example: Preparation of Extract]

1. Preparation of Hemp Extract

Hemp, including leaves and flowers, was thoroughly washed under running water and then completely air-dried. The dried hemp was pulverized with a blender and then prepared into powder. 50% ethanol as an extraction solvent was added to the powder sample at a ratio of 1;10 (w; v), and then completely immersed, and extracted three times for 3 hours while refluxing at 80°C. The extract is Whatman No. 2 filtered with filter paper. The filtrate was concentrated under reduced pressure at 60° C. to prepare a hemp extract (CE).

2. Manufacture of other natural extracts

By using the same method as the method for preparing the hemp extract (CE), the annual extract (EE), the carob extract (IE) and the star flower extract (SE) were prepared.

3. Preparation of Complex Extracts

The cannabis extract (CE), yilnyeonbong extract (EE), carob extract (IE) and starflower extract (SE) were mixed as shown in Table 1 below to obtain a complex extract.

MX1 MX2 MX3 MX4 MX5 MX6 CE 100 100 100 100 100 100 EE - 10 20 30 40 50 IE - 10 20 30 40 50 SE - 10 50 30 40 50

(Unit: parts by weight)

[Experimental Example 1: Cytotoxicity test]

In order to test the toxicity of the hemp extract (CE) and the complex extract (MX2 to MX6), the difference in toxicity and side effects expression was confirmed when the complex extract was administered in a repeated dose toxicity test in rats.

Six-week-old male and female rats of SD opening were divided into 10 rats per group (5 males and females each) and complex extracts MX1 to MX6 were administered. This was repeated for 13 weeks.

A single dose was administered in an amount of 3.75 mg/kg to 5 mg/kg. Thereafter, mortality, general symptoms, weight change, and feed and water intake were observed.

As a result, no deaths occurred within the experimental period. In light of the above experimental results, it was confirmed that the hemp extract (CE), the annual extract (EE), the carob extract (IE), the star flower extract (SE) and mixtures thereof do not have any toxicity problems.

[Experimental Example 2: Pain Relief Effect]

1. Cyclooxygenase (COX)-2 inhibitory ability

Hemp extract (CE), a sample to be used in the cyclooxygenase (COX)-2 enzyme inhibition test, was used in DMSO solution. First, COX-2 enzyme, heme, and an inhibitory sample were put into a test tube containing the reaction buffer (control group, put the same amount of DMSO instead of the inhibitory sample), and then reacted at 37°C for 10 minutes. After 10 minutes of reaction, 10 μL of substrate (arachidonic acid) was added and the reaction was continued for 2 minutes. After the reaction was completed, 50 μL of 1M HCl solution was added to terminate the reaction, and then tartaric acid was added to cause a reduction reaction. After 2000-fold and 4000-fold dilution of each test tube reaction solution, 50 μL each was placed in a 96-well plate. After adding 50 μL of tracer and antiserum, respectively, react at 25° C. for 18 hours. After the reaction was completed, the wells were emptied, rinsed 5 times using a washing buffer, and then a color reagent was added and reacted for 1 hour and 30 minutes. When the reaction was completed, the absorbance was measured at 405 nm using an ELISA reader.

As a result, the inhibitory ability of the hemp extract (CE) on COX-2 was shown in Table 2. When each sample contained the same level of hemp extract (CE) at the same level (1 mg/mL), 35.75% of COX-2 enzyme inhibitory activity was shown, whereas MX2 to MX6 were 58.57%, 60.38%, 62.47%, and 61.97%, respectively. , showed an inhibitory activity of 60.27%. The complex extracts (MX2 to MX6) showed an inhibitory activity of about 1.7 times or more compared to MX1, a hemp extract (CE).

Therefore, as a result of inhibiting the COX-2 enzyme, which is a synthesizing enzyme of prostaglandin (PG), it was confirmed that the composite extracts (MX2 to MX6) at the same concentration had a superior inhibitory effect compared to the cannabis extract.

COX-2 Inhibition MX1 35.75±2.45 ¹ MX2 58.57±8.08 ² MX3 60.38±5.41 ² MX4 62.47±2.41 ² MX5 61.97±9.06 ² MX6 60.27±4.41 ²

(unit: %)

In addition, as a result of checking the complex extracts MX3 to MX5 showing the highest inhibitory effect, it was confirmed that the cyclooxygenase (COX)-2 inhibitory effect was excellent in this range.

2. Formalin Pain Model Experiment of Hemp Extract (CE)

TMJ (temporomandibular joint) model (temporomandibular joint pain model, Won KA, Kang YM, Lee MK, Park MK, Ju JS, Bae YC, et al. Participation of microglial p38 MAPK in formalin-induced temporomandibular joint nociception in rats. J Orofac Pain 2012; 26(2): 132-141.) was used to conduct an acute temporomandibular joint pain relief experiment on hemp extract (CE).

To explain the specific experimental method, the experimental animals were acclimatized for at least 10 minutes in an experimental plastic container before pain response evaluation. 30 μl of 5% formalin was injected using an insulin syringe (0.25 × 8 mm), and it was observed that consciousness was restored within a few seconds after formalin injection.

The position of the joint cavity was inferred by palpation of the posterior inferior interface of the zygomatic arch and the mandibular condyle. Through a preliminary experiment, the position of the temporomandibular joint cavity was confirmed by injecting the same amount of 1w% evans blue dye as formalin into a separate animal. After formalin injection, the reaction of rubbing or scratching the TMJ area and the facial area was regarded as a pain index and accumulated for 45 minutes 9 consecutive times at 5-minute intervals, and the primary pain behavior response (0 to 10 minutes) and secondary pain behavior response (11 to 45 minutes) was evaluated separately.

As a result, it was found that in an experiment using animals (Rat), a change in the pain behavior response appeared when the hemp extract (CE) was administered in the formalin-induced acute pain model administered to the temporomandibular joint. As for the primary pain behavior response, the pain behavior response to which the hemp extract was administered was not significantly different between the formalin injection group, the control group (veh + formalin injection group) and the hemp extract (CE) injection group (30㎛, 50㎛, 100㎛). However, it was effective in controlling pain at 20 to 35 minutes, which corresponds to the secondary pain behavior response. These results are shown in FIG. 1, and it can be seen that the cannabis extract (CE) has a pain relief effect.

3. Formalin Pain Model Experiment of Complex Extracts

In order to examine the pain relief effect of the complex extracts (MX2 to MX6), the same experiment as that of the hemp extract (CE) was conducted, and the results were compared and shown as a relative effect on the hemp extract (CE).

For the pain relief effect compared to the hemp extract (CE), CE was set as an index of 5, and the temporomandibular joint pain relief of MX2 to MX6 was evaluated as an index 1 to 10. It will be said that the higher the index, the better the pain relief effect.

CE MX2 MX3 MX4 MX5 MX6 temporomandibular joint pain model 5 6 7 8 7 6

(Unit: Index)

As shown in Table 3, compared to the hemp extract (CE), it was confirmed that the complex extracts MX2 to MX6 relieved pain equal to or more than that. This will mean that, compared to the case of using only the hemp extract (CE), when used as a natural complex extract, the effect of reducing pain due to the complex action between each component is excellent.

[Experimental Example 3: Stress prevention and improvement effect]

Four-week-old male ICR mice (Samtaco Bio Korea, Korea) were subjected to an adaptation period for 1 week, then the lighting was turned on and off repeatedly every 12 hours, and the room temperature was maintained at 18 to 23°C and humidity of 60%. bred in a space where As the feed, solid feed was supplied, and there were no restrictions on feed and water supply except for the process of inducing stress.

After dissolving the complex extracts (MX1 to MX6) of Preparation Example in distilled water, it was orally administered (40 mg/kg) once daily at 14-15 o'clock for one month continuously. The experimental group consisted of a total of three groups, a control group given only stress, a group administered with complex extracts (MX1 to MX6), and a normal control group that received neither stress nor extract. For each experimental group, 5 animals were used, and stress was induced every day for 4 weeks from 7 days after administration.

The stress-inducing method is used by modifying the method of Willner et al. (Wilner et al., Reduction fo sucrose preference by chronic unpredictable mild stress, 1987). Specifically, fasting, dietary restriction after fasting (feeding a small amount), single , providing an empty water bottle after a water supply, a slanted kennel, breeding a large number of experimental animals in one kennel, and creating various unexpected psychological stressful situations such as flashing lights, cold rooms, constant lights, etc., induce stress.

After the stress-induced test animal was sacrificed by cervical dislocation, the brain was removed and the hippocampal tissue was placed in a cooled tube and rapidly cooled. Each tissue sample was treated with 0.4 M perchloric acid and sonicated, followed by centrifugation at 12,000 rpm (4° C.) in a centrifuge for analysis. For analysis, a high performance liquid chromatography with electrochemical detection (HPLC-ECD) method was used with a slight modification of the method of Qi et al.

1. Dopamine inhibitory effect

Dopamine is a neurotransmitter in the central nervous system, a precursor to norepinephrine, and is involved in cognition, attention, concentration, reward, and motor function regulation. Since it is known that dopamine secretion is suppressed in a chronic stress situation, the stress prevention and improvement effect of the composition of the present invention was confirmed through a dopamine measurement experiment.

The results of measuring the dopamine concentration using the high performance liquid chromatography with electrochemical detection (HPLC-ECD) method were compared with the dopamine concentration of the normal control group (% of control) as the mean and standard error. Significance was verified by statistical treatment using Student's T-test, and the group treated with cannabis extract (CE, MX1), the group administered with complex extracts (MX2 to MX6) and the stress control group were compared and displayed as significant at p<0.05. The results are shown in FIG. 2 .

Referring to FIG. 2 , it can be seen that the concentration of dopamine measured in the hippocampus of the stressed test animal is significantly lower than that of the normal control group. However, it can be seen that the concentration of dopamine was maintained higher in the group administered with the hemp extract (CE) and the complex extract (MX2 to MX6) compared to the stress control group.

In particular, compared to the case of using only the hemp extract (CE, MX1), when using the natural complex extracts MX3 to MX5, it will mean that the effect of inhibiting the decrease in dopamine by the complex action between each component is excellent.

Therefore, it can be seen that when according to the above range, the effect of preventing and improving stress by suppressing the decrease of dopamine due to stress is exhibited.

2. Serotonin inhibitory effect

The results were compared with the serotonin concentration of the normal control group (% of control) and expressed as the mean and standard error. Significance was verified by statistical treatment using Student's T-test, and the group treated with cannabis extract (CE, MX1), the group administered with complex extracts (MX2 to MX6) and the stress control group were compared and displayed as significant at p<0.05. The results are shown in FIG. 3 .

Referring to FIG. 3 , it can be seen that the serotonin measured in the hippocampus of the stressed test animal is significantly lower than that of the normal control group. However, it can be seen that the serotonin concentration was maintained high in the group administered with the hemp extract (CE) and the complex extract (MX2 to MX6) compared to the stress control group.

In particular, compared to the case of using only the hemp extract (CE, MX1), when using the natural complex extracts MX3 to MX5, it will mean that the effect of inhibiting the decrease of serotonin by the complex action between each component is excellent.

Therefore, it can be seen that when according to the above range, it can be seen that the reduction of serotonin due to stress is suppressed to prevent and improve stress.

[Experimental Example 4: Taste Test]

1. Sensory evaluation test

MX1 and complex extracts MX3 to MX6 were diluted to prepare a tea beverage. The tea drink was tasted by 10 tasters, and the taste and aroma were expressed as an index of 1 to 10, and the average value (0.5 rounding applied) is shown in Table 4 below. In the index, the higher the number, the higher the degree of preference.

MX1 MX2 MX3 MX4 MX5 MX6 taste 6.0 6.0 6.5 7.0 7.0 6.0 incense 6.0 6.5 6.5 7.0 7.5 7.0 Overall preference (average) 6.0 6.0 7.0 7.0 7.0 6.5

(Unit: Index)

Referring to Table 4, in the case of MX1, hemp extract was used alone to lower the taste and flavor, and in the case of mixing MX2 to MX6, the unique taste and flavor of the hemp extract was neutralized by other natural extracts. It can be seen that the degree of preference increases. In particular, it was confirmed that in the case of MX3 to MX5, the preference was greatly increased while the fragrance was highly evaluated.

Therefore, in the case of the complex extracts MX3 to MX5, it is possible to provide a functional food having excellent pain relief and stress-related disease prevention and improvement effects with a higher preference for flavor and taste.

Although the preferred embodiment of the present invention has been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements by those skilled in the art using the basic concept of the present invention as defined in the following claims are also provided. is within the scope of the

Claims (9)

Contains hemp (Cannabis sativa L.) extract as an active ingredient
A composition for relieving pain and preventing and improving stress-related diseases. The method of claim 1,
The hemp (Cannabis sativa L.) extract comprising cannabinoids (cannabinoids) and terpenes (terpene)
A composition for relieving pain and preventing and improving stress-related diseases. The method of claim 1,
The stress-related disease is any one selected from the group consisting of stress, depression and insomnia
A composition for relieving pain and preventing and improving stress-related diseases. The method of claim 1,
The composition inhibits the cyclooxygenase (COX)-2 enzyme to exhibit a pain relief effect
A composition for relieving pain and preventing and improving stress-related diseases. The method of claim 1,
The composition is to prevent dopamine and serotonin from being inhibited
A composition for relieving pain and preventing and improving stress-related diseases. The method of claim 1,
The extract is extracted using an extraction solvent selected from the group consisting of water, C ₁ to C ₆ lower alcohols and mixtures thereof.
A composition for relieving pain and preventing and improving stress-related diseases. A functional food comprising the composition for alleviating pain and preventing and improving stress-related diseases according to any one of claims 1 to 6. [Claim 7] An external preparation for skin comprising the composition for alleviating pain and preventing and improving stress-related diseases according to any one of claims 1 to 6. A pharmaceutical comprising the composition for relieving pain and preventing and improving stress-related diseases according to any one of claims 1 to 6.

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