WO2014175518A1 - Composition for relieving, preventing, or treating pain containing mate extract as active ingredient, and method for evaluating pain relief, prevention, or treatment efficacy of sample - Google Patents

Abstract

The present invention provides a composition for relieving, preventing, or treating pain containing a mate extract as an active ingredient. The composition of the present invention has effects of relieving and treating pain that has occurred, and exhibits an effect of preventing pain when being administered to a subject before the pain occurs. The composition of the present invention exhibits a remarkable pain relief effect even through oral administration as well as through intravenous injection, application to skin, or the like. The composition of the present invention having these features is highly applicable to a food. The present invention provides a method for evaluating pain relief, prevention, or treatment efficacy of a sample. The use of the present invention can accurately measure the number of times of occurrence or the duration time of ultrasonic vocalization generated from an experimental animal on pain response and express the measurement results as objective numerical values, so that the present invention is very useful in evaluating pain relief, prevention, or treatment efficacy of the sample, and is capable of minimizing errors of the prior art resulting from individual differences among experimenters and subjective judgments in the measurement of the pain response.

Description

 【Specification】

[Name of invention]

 Evaluation method for the efficacy of pain relief, prevention or treatment of the composition for the relief, prevention or treatment of pain and the sample containing mate extract as an active ingredient

Technical Field

 The present invention relates to a composition for the relief, prevention or treatment of pain, comprising a mate extract as an active ingredient, and a method for evaluating the efficacy of pain relief, prevention or treatment of a sample.

[Related Patent Application]

 This patent application claims priority to Korean Patent Application No. 10-2013-0044943 and Korean Patent Application No. 10-2013-0044944 filed with the Korean Intellectual Property Office on April 23, 2013. Is incorporated herein by reference.

Background Art

 Currently, as analgesics, anti-inflammatory drugs such as aspirin and tylenol are the mainstream, and in case of severe pain, morphine drugs are mostly used. Recently, a new pain inhibitor is being developed. Javelin Pharmaceuticals, a developer specializing in pain medications, has introduced a nasal morphine spray, a nasal morphine spray that has an analgesic effect equivalent to intravenous morphine for the treatment of pain after severe surgery. 'Rylomine' was developed. In addition, the painkiller `` Prialt '', developed by Irish pharmaceutical company Elan from sea snail poison, was first marketed in the UK.

As the society ages as well as chronic malignant neoplastic pain, the number of patients with degenerative arthritis and low back pain is increasing every year. However, the use of existing opiates such as morphine is restricted due to narcotic action. It is expected to be much longer. In addition, there is a great demand for the development of painkillers for pain that does not respond to existing painkillers. Due to the narcotic and serious side effects of conventional opioids, which are known to have relatively large analgesic effects, the general patient cannot use them comfortably. Therefore, there is a need for the development of natural resources having no side effects and having analgesic efficacy.

 Mate (Ilex paraguariensis, mate) is an evergreen subfamily of persimmon tree, which is 6 m tall and is distributed in Paraguay and Brazil in South America. In the wild state, the upper part is in the shape of the back, but when it is grown, its shape is modified to become a shrub with many stems, and it is possible to grow tea leaves of higher quality than the wild state, and harvesting is performed every two years. The flowers of mate are small, hang on the axilla, 4 digits, green, and the fruit is red or reddish brown, orange, and 6 지름 in diameter. Mate leaves have been a favorite drink of Latin Americans for thousands of years and are a source of tea with aroma similar to green tea, Thea sinensis L. (Wikipedia, 2011). The leaves contain caffeine and have a good aroma (around 2%), and the leaves are harvested and dried for drinking as tea.

 Mate grows in mineral-rich soils such as calcium and iron, so it contains a lot of minerals such as iron scabbard, especially iron, and has an iron content that is five times higher than that of green tea. In particular, other teas contain tannins that interfere with iron absorption, while mate contains little tannin. Mate tea is made from dried leaves of mate and contains triterpene, caffeine and caffeine-like compounds. Mate tea is known to be effective in the treatment of constipation, obesity, shoulder stiffness, headache, etc. The pharmacological activity of these mate is due to caffeoyl quinic acid and caffeine-like polyphenols. The main components of Mate are mainly contained in large amounts of flavoids such as chlorogenic acid, rutin, quercetin, etc. (SONIA CHANDRA et al., 2004), antioxidant and anti-inflammatory action, blood vessel relaxation and blood lipid suppression, anticancer action, hypoglycemic activity Various pharmacological effects are known, such as action, weight loss efficacy, etc. (Bracesco et al., 2011).

Previous studies on mate have found the presence of many bioactive compounds such as xanthine, caffeoylerivative, flavonoids and tri terpenoid saponins. Yerba mate (ilex paraguariensis) is an antioxidant and Cell protective action (Filip et al., 2000; Gugliucci and Stahl, 1995), anti-obesity and pyrogenic action (Martinet et al., 1999), circulatory action and lowering cholesterol (Gorgen et al., 2005) and bile A variety of biological activities are known, such as promoting action (Gorzalczany et at., 2001).

In order to measure the pain level of the experimental animals, mechanical allodynia evaluation (von frey filament test), weight bearing test (Weight bearing test) is mainly used. Since these conventional test methods rely on subjective judgment of the experimenter, a problem arises that the accuracy of the experiment is degraded in many cases. Therefore, careful blind test is required. In addition, the conventional test method has a problem that it is difficult to obtain a good quality test data because the experimenter has to repeatedly test for a long time using the arm is labor-intensive and takes a lot of test time. Thus, the experimental animal ^'objective and easy techniques that can accurately measure the pain in banung is required. Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

[Content of invention]

 Problem to be solved

The present inventors have found that a safe substance for a human body that can effectively alleviate ⁱ pain, in particular plant-was intensive studies seek to develop derived materials, and as a result to the mate that is being used as the primary raw material in the conventional relief, preventing or treating pain in a very It was found to be valid.

In addition, the present inventors made extensive efforts to develop a method for evaluating the pain relief, prevention, or treatment efficacy of a sample, and as a result, after administering a sample to be analyzed to a pain-causing animal, 22 The present invention was completed by elucidating that measuring ultrasound at -27 kHz is very effective in evaluating the efficacy of pain relief, prevention or treatment of a sample. Accordingly, an object of the present invention is to provide a composition for preventing or treating pain relief.

 Another object of the present invention to provide a food composition and pharmaceutical composition for the relief, prevention or treatment of pain.

 Another object of the present invention to provide an evaluation method for the efficacy of pain relief, prevention or treatment of a sample.

 Other objects and advantages of the present invention will become apparent from the following detailed description and claims. [Task solution]

 According to an aspect of the present invention, the present invention provides a composition for alleviating, preventing or treating pain comprising an extract of Mate paraguayensis as an active ingredient. '

 The present inventors have made intensive studies to develop substances that are safe for humans, especially plant-derived substances, which can effectively alleviate pain, and as a result, Mate, which is conventionally used as a raw material for tea, is very effective in alleviating, preventing or treating pain. It was identified.

 The composition of the present invention works very effectively in alleviating, preventing or treating pain.

 As demonstrated in the following examples, when the composition of the present invention was administered to an animal prior to skin incision surgery, the pain sensitivity at the surgical site was decreased and the composition of the present invention was administered to an animal with nerve branch ligation injury. , Pain sensitivity decreased.

 The composition of the present invention exhibits a significant pain relief effect through oral administration rather than intravenous injection or skin application. The composition of the present invention having these characteristics can be well applied to food.

The composition of the present invention not only has a palliative and therapeutic effect on the previously generated pain, but also has an effect of preventing pain when administered to a subject before the pain occurs. Mate (Ilex paraguayensis) used as an active ingredient in the composition of the present invention is a plant belonging to the persimmon tree as an evergreen subfamily of persimmon tree.

 When the mate extract used in the composition of the present invention is obtained by treating the mate extractant, various extractive solvents may be used. According to the present invention, a polar solvent or a nonpolar solvent can be used. Suitable polar solvents include (i) water, (ii) alcohols (preferably methanol, ethanol, propane, butanol, normal-propane, iso-propane, normal-butane, 1-pentane, 2 -Butoxyethane or ethylene glycol), (iii) acetic acid, (iv) dimethylformamide (DMFO) and (v) dimethyl sulfoxide (DMS0). Suitable as nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, nucleic acid, 2, 2,4-trimethylpentane, decane, cyclonucleic acid, cyclopentane, diisobutylene, 1- Pentene, 1-chlorobutane, 1-chloropentane, 0-xylene, diisopropyl ether 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloro Methane, 1,2-dichloroethane, anneal, diethylamine, ether, carbon tetrachloride and THF.

 According to one embodiment of the present invention, the extraction solvent used in the present invention is (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methane, ethane, propane, butanol, etc.), ( c) a mixed solvent of the lower alcohol and water, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) butyl acetate (h) 1,3-butylene glycol, (i) hexane and (j) diethyl ether. In another embodiment of the present invention, the extraction solvent used in the present invention is water, ethanol or a combination thereof. According to a particular embodiment of the present invention, the extraction solvent used in the present invention is 4 ()-80 vol% grain alcohol.

 According to one embodiment of the present invention, the mate extract of the present invention is obtained by adding alcohol to the mate leaves, followed by filtration and concentration under reduced pressure, followed by lyophilization.

Mate extract used in the present invention may be prepared in a powder state by an additional process such as vacuum distillation and freeze drying or spray drying. As used herein, the term 'extract' has the meaning commonly used as a crude extract in the art as described above, but also broadly includes a fraction additionally extracting the extract. That is, the mate extract is not only obtained by using the above-described extraction solvent, but also includes one obtained by additionally applying a purification process. 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 (manufactured for separation according to size, charge, hydrophobicity or affinity), etc. The fraction obtained by the method is also included in the mate extract of the present invention.

As used herein, the term 'comprising as an active ingredient' means including an amount sufficient to achieve the efficacy or activity of the following Mate extract. Embodiment the invention composition is selected in overdose and also because there is no adverse effect on the human body comprising the composition of the invention is Mate extract ^present, the quantity upper limit is the range one of ordinary skill in the art is suitable as comprising an extract obtained from the mate of the natural plant material can do.

 The term "pain" is used herein in a broad sense and includes acute and chronic pain, such as somatic pain, such as somatic pain and visceral pain; Inflammatory pain, dysfunction pain, idiopathic pain, superficial pain, deep pain, itching, neuropathic pain, e.g., centrally generated pain, and peripheral pain (peripherally generated pain), all types of pain including migraine and cancer pain.

The term “nociceptive pain” is used to encompass all pain caused by harmful stimuli that may damage or actually damage body tissues, including cuts, bruises, Pain without bone fracture, crush injury, burn and similar wounds, without limitation. Pain receptors for tissue damage (nociceptors) are mostly located in the skin, musculoskeletal system or internal organs. The term “somatic pain” is used to refer to pain that occurs in bones, joints, muscles, skin or connective tissue. This type of pain is typically very local.

 The term “visceral pain” is used herein to refer to respiratory, gastrointestinal tract and pancreas in intestinal organs, urinary tract and reproductive organs. Used to indicate pain that occurs. Visceral pain includes pain induced by tumor involvement of the organ capsule. Other types of visceral pain are typically characterized by intermittent cramping and severe local pain caused by obstruction of the hoi low viscus. Visceral pain may be associated with inflammation as in the case of cystitis or reflux esophagit is.

 The term "inflammatory pain" includes pain associated with active inflammation that may be caused by trauma, surgery, infection, autoimmune diseases, and the like.

 The term "superficial pain" refers to pain felt along the skin segment in which the nerves of the dorsal root are distributed, and is a direct pain that feels pain at the point of stimulation.

The term "deep pain" refers to pain originating from deep organs and differs in the nature and extent of pain depending on the nature of the tissue. Particularly sensitive to pain are tendons, deep muscles, ligaments, joints, periosteum, blood vessels and nerves. In general, deep pain is a sensation that is dull and spread around. Deep and internal pain, such as the complex mechanism of the pain is difficult to find the location of the pain, nausea, sweating, increased blood pressure also appears. The term "neuropathic pain" is used herein to refer to pain resulting from sensory input of abnormal processing by the peripheral or central nervous system resulting from peripheral or central nervous system disorders. The term "procedural pain" refers to pain that occurs in a medical, dental or surgical procedure, which procedure is usually scheduled or associated with acute trauma.

 The term "itch" is used herein in a broad sense and refers to all types of itchy and stinging sensations of acute intermittent and persistent, which can be generally explained locally. The itching may be idiopathic, allergic, metabolic, infectious, drug-induced, liver, kidney disease or cancer. "Pruritus" is severe itching.

 "Patient" means any animal. In one embodiment of the invention, the patient is a human. Other animals that can be treated using the compositions of the present invention include non-human primates (eg, monkeys, gorillas, chimpanzees), livestock (eg, horses, pigs, goats, rabbits, sheep, cattle, llamas), and companion dogs. Animals such as but not limited to guinea pigs, rats, mice, lizards, snakes, dogs, cats, ornamental fish, hamsters and birds.

 Neuropathic, inflammatory, and nociceptive pain differ in etiology, pathophysiology, diagnosis, and treatment. Nociceptive pain occurs in response to the activation of nociceptors by specific subsets of peripheral sensory neurons, i.e., intense or noxious stimuli. Nociceptive pain. In general, it provides a biological function of protection by acting as a warning of sensitive, self-limiting, and potential or ongoing tissue damage. Nociceptive pain is typically very limited locally. Examples of nociceptive pain include traumatic or surgical pain, labor pain, sprains, bone fractures, burns, bumps, bruises ( bruises, injections, dental procedures, skin biopsies, and obstructions.

Inflammatory pain is postoperative, post-traumat ic pain, arthritis (rheumatoid or osteoarthritis) Pain, and pain that occurs when there is tissue damage or inflammation, including pain associated with damage to joints, muscles, and tendons, such as in the case of axial low back pain.

 In contrast to nociceptive pain, neuropathic pain is actually described as "burning," "electric," "tingling," or "shooting." do. Neuropathic pain is often defined as chronic al lodynia (defined as pain due to irritation that usually does not cause pain response, such as a light touch) and hyperalgesia (high sensitivity to normal pain stimulation). And may last for months or years after apparent healing of any damaged tissue.

 According to one embodiment of the present invention, the composition of the present invention works very effectively to alleviate, prevent or treat persistent peripheral neuropathic pain.

 The composition of the present invention may be provided as a food composition.

 When the composition of the present invention is prepared as a food composition, it contains not only mate extract as an active ingredient, but also components commonly added in food production, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors. Include the first. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, loli sugar and the like; And sugars such as polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and xyls, sorbitols, and erythrates. As flavoring agents, natural flavoring agents (tauumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

 For example, when the food composition of the present invention is manufactured with a drink, in addition to the mate extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. may be further included. have.

The composition of the present invention may be provided in a pharmaceutical composition. When the composition of the present invention is prepared as a pharmaceutical composition, it contains not only mate extract as an active ingredient, but also a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are those commonly used in the preparation of lactose, dextrose, sucrose, sorbbi, manny, starch, acacia rubber, phosphate chalc, alginate, Gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyridone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil Including, but not limited to. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

 The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like.

 Suitable dosages of the pharmaceutical compositions of the present invention vary depending on factors such as the formulation method, mode of administration, age of the patient, weight, sex, morbidity, food administration time, route of administration, rate of excretion and response sensitivity. Usually a skilled practitioner can easily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dose of the pharmaceutical composition of the present invention is 0.001-100 mg / kg.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, or emulsion in an oil or an aqueous medium, or may be in the form of axes, powders, granules, tablets, or accelerators, and may further include a dispersant or stabilizer. According to another aspect of the present invention, the present invention provides a method for evaluating the efficacy of pain relief, prevention or treatment of a sample comprising the following steps:

 (a) causing pain in animals other than humans;

 (b) administering a sample to be analyzed to the animal; And

 (c) measuring 22-27 kHz ultrasound generated from the animal to which the sample was administered, wherein the sample reduces pain when the sample reduces the 22-27 kHz ultrasound generated from the animal; It is determined to have a prophylactic or therapeutic effect.

 The present inventors made intensive studies to develop a method for evaluating the pain relief, prevention or treatment efficacy of a sample. As a result, after administering a sample to be analyzed to the pain-induced animal, 22-27 Ultrasound measurements at kHz were found to be very effective in evaluating the pain relief, prophylactic or therapeutic efficacy of a sample.

 According to the method of the present invention, after causing the pain in the animal, the sample to be analyzed is administered and the number or duration of ultrasound generation of 22-27 kHz generated from the animal to which the sample is administered is measured. The frequency or duration of the 22-27 kHz lunar zone generated from the animal was measured and compared to the animal that did not receive the sample by measuring the frequency or duration of the ultrasonic When reduced, the sample is considered to be a substance for alleviating or treating pain.

 According to one embodiment of the invention, the step of administering the sample to be analyzed in the animal may be carried out first before causing the pain in the animal. After the sample to be analyzed is administered to the animal prior to pain, the number or duration of the ultrasound generation of 22-27 kHz from the animal is measured and compared with the animal that has not been given the sample. The sample is considered to be a material for the prevention of pain when the frequency or duration of ultrasound generation at kHz decreases.

In the present invention, the sample may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like. The sample used in the present invention is a peptide, an antibody, a peptide aptamer, AdNectin, affibody, US Patent No. 5,831,012, Avimer (Avimer, Silverman) , J. et al, Nature Biotechnology 23 (12): 1556 (2005)) or Kunitz domain, Arnoux B et al., Acta Crystal logr.D Biol.Crystal logr. 58 (Pt 7): 1252- 4 (2002)), and Nixon, AE, Current opinion in drug discovery & development 9 (2): 261-8 (2006).

 Pain induction in animals in the present invention can be induced through drug administration, skin incision or nerve ligation. Pain-induced animals can use the pain animal model proposed by various pain-inducing methods known in the art. For example, the pain animal model used in the present invention may include a CCKChronic constriction injury model, a PSL / partial sciatic nerve ligation model, a spinal nerve ligation model, a split nerve injury model, a Vincrist in Vincrist in —Induced neuropathy model, Incision model, intradural catheterization model, direct lumbar puncture model.

 According to one embodiment of the invention, the animal used in the evaluation method of the present invention is a rodent animal.

 According to certain embodiments of the invention, the animal used in the evaluation method of the present invention is a mouse (Mus musculus) or a rat (Rattus norvegicus). '

 The term "pain" is used in a broad sense in the evaluation method for the efficacy of pain relief, prevention or treatment of a sample of the present invention, and acute and chronic pain, such as somatic pain, for example somatic pain and visceral. Visceral pain; Inflammatory pain, dysfunction pain, idiopathic pain, superficial pain, deep pain, itching, neuropathic pain, such as centrally generated pain and peripheral pain (peripherally) generated pain, migraine and cancer pain.

The term "nociceptive pain" is used to encompass all pain caused by harmful stimuli that may damage or actually damage body tissues, such as cuts, bruises, Pain without bone fracture, crush injury, burn and similar wounds, without limitation. Pain receptors for tissue damage (nociceptors) are mostly located in the skin, musculoskeletal system or internal organs.

 The term "somatic pain" is used to refer to pain that occurs in bones, joints, muscles, skin or connective tissue. This type of pain is typically very local.

The term "visceral ^' pain" is used herein to refer to organs such as the respiratory, gastrointest inal tract and pancreas, urinary tract and reproductive organs. It is used to indicate pain occurring in). Visceral pain includes pain induced by tumor involvement of the organ capsule. Other types of visceral pain are typically caused by obstruction of the hollow viscus and are characterized by intermittent cramping and severe local pain. Visceral pain may be associated with inflammation as in the case of cystitis or reflux esophagi t is.

 The term "inflammatory pain" includes pain associated with active inf la 隱 at ion, which may be caused by trauma, surgery, infection, autoimmune diseases, and the like.

 The term "superficial pain" refers to pain felt along the skin segment where the nerves of the dorsal root are distributed, and is a direct pain that feels pain at the point of stimulation.

The term "deep pain" refers to pain originating from deep organs and differs in the nature and extent of pain depending on the nature of the tissue. Particularly sensitive to pain are tendons, deep muscles, ligaments, joints, periosteum, blood vessels and nerves. In general, deep pain is a sensation that is dull and spread around. Deep and internal pain, such as the complex mechanism of the pain is difficult to find the location of the pain, nausea, sweating, increased blood pressure also appears. The term "neuropathic pain" is used herein to refer to the peripheral or central nervous system as a result of peripheral or central nervous system disorders. It is used to represent pain resulting from sensory input of abnormal processing.

 The term "procedural pain" refers to pain that occurs in a medical, dental or surgical procedure, which procedure is usually scheduled or associated with acute trauma.

 The term "itch" is used herein in a broad sense and refers to all types of itchy and stinging sensations of acute intermittent and persistent, which can generally be explained locally. The itch may be idiopathic, allergic, metabolic, infectious, drug-induced, liver, kidney disease or cancer. "Pruritus" is severe itching.

 "Patient" means any animal. In one embodiment of the invention, the patient is a human. Other animals that can be treated using the compositions of the invention include non-human primates (eg, monkeys, gorillas, chimpanzees), livestock (eg, horses, pigs, goats, rabbits, sheep, cattle, llamas), and companion dogs. Animals, such as, but not limited to, guinea pigs, rats, mice, lizards, snakes, dogs, cats, ornamental fish, hamsters, and birds.

Neuropathic, inf la 薩 atory, and nociceptive pain differ in etiology, pathophysiology, diagnosis, and treatment. Nociceptive pain occurs in response to the activation of nociceptors by specific subsets of peripheral sensory neurons, i.e. intense or noxious stimuli. Nociceptive pain generally provides a biological function of protection by acting as a warning of sensitive, self-limiting and potential or ongoing tissue damage. Nociceptive pain is typically very limited locally. Examples of nociceptive pain include traumatic or surgical pain, labor pain, sprains, bone fractures, burns, bumps, bruises ( bruises, injections, dental procedures, skin biopsies, and obstructions. Inflammatory pain may be associated with joints such as postoperative, post-traumat ic pain, arthritis (rheumatoid or osteoarthritis) pain, and axial low back pain. pain that occurs when there is tissue damage or inflammation, including pain associated with damage to joints, muscles, and tendons.

 In contrast to nociceptive pain, neuropathic pain is actually "burning," "electric," "tingling," or "shooting." Is depicted. Neuropathic pain is often defined as chronic al lodynia (defined as pain due to irritation that usually does not cause pain, such as a light touch) and hyperalgesia (high sensitivity to normal pain stimulation). And may last for months or years after apparent healing of any damaged tissue. 【effect】

 The features and advantages of the present invention are summarized as follows:

 (a) The present invention provides a composition for alleviating, preventing or treating pain comprising a mate extract as an active ingredient.

 (b) The composition of the present invention not only has a palliative and therapeutic effect on the previously generated pain, but also has an effect of preventing pain when administered to a subject before the pain occurs.

 (c) The composition of the present invention shows a significant pain relief effect through oral administration other than the method of intravenous injection or skin application, the composition of the present invention having such characteristics is highly applicable to food.

 (d) The present invention provides a method for evaluating the efficacy of pain relief, prevention or treatment of a sample.

(e) By using the present invention can accurately measure the number and duration of the ultrasonic lunar zone generated during pain reaction from the experimental animal can be expressed as an objective value, very useful for evaluating the pain relief, prevention or treatment efficacy of the sample. (f) The present invention can minimize errors in subjective judgments and individual differences of experimenters in the measurement of pain response.

[Brief Description of Drawings]

 1 is a result of oral administration of the mate extract to the experimental animals, and 5 hours after the skin incision surgery, the pain sensitivity of the surgical site was measured through mechanical allodynia evaluation.

 Control group (n = 10), mate extract (300 mg / kg) administration group (n = 10), * p <0.05 Figure 2 shows the oral administration of mate extract to the experimental animals, the skin incision surgery was performed 24 hours after surgery The pain sensitivity of the site was measured by mechanical allodynia evaluation.

 Control group (n = 10), mate extract (300 mg / kg) administration group (n = 10), * p <0.05 Figure 3 is an oral administration of mate extract to the experimental animals, and then performed the skin incision 6 hours after the experiment This is a measure of the number of times an animal generates ultrasound at the 22–27 kHz lunar calendar.

 Control group (n = 10), mate extract (300 mg / kg) administration group (n = 10), * p <0.05 Figure 4 is an oral administration of mate extract to the experimental animals, the skin incision surgery 24 hours after the experiment This is the result of measuring the number of times an animal generates ultrasonic waves in the 22-27 kHz lunar calendar.

 Control (n = 10), Mate extract (300 mg / kg) administration group (n = 10), * p <0.05 Figure 5 is the result of measuring the pain threshold value of experimental animals before nerve branch ligation injury surgery.

 Control group (n = 10), mate extract (300 mg / kg) administration group (n = 10)

 6 is a result of measuring the pain threshold value after oral administration of mate extract to experimental animals for 14 days after nerve branch ligation injury surgery.

 7 is a diagram showing the manufacturing process of the skin incision model (acute pain model).

 8 is a diagram of a method for producing a nerve branch ligation injury model (chronic pain model).

9 is a diagram showing the manufacturing process of the nerve branch ligation injury model. 10 is a diagram showing a mechanical allodynia evaluation method. 11 is a diagram illustrating a method for measuring ultrasonic utterance sound. 12 is a diagram illustrating a result of ultrasonic sound measurement.

 (a) control group, (b) pain relief treatment group

 Figure 13 is a result of measuring the pain sensitivity of the surgical site by mechanical allodynia after 5 hours after intraperitoneal injection of gabapentin to the experimental animals immediately after the skin incision surgery.

 Control group (n = 10), gabapentin (30 mg / kg) administration group (n = 10), *** ρθ.001 Figure 14 shows the intraperitoneal injection of gabapentin into the experimental animals immediately after skin incision surgery, P0D1) The pain sensitivity of the surgical site was measured by mechanical allodynia evaluation.

 Control group (n = 10), gabapentin (30 mg / kg) administration group (n = 10), * p <0.05 Figure 15 shows that after the intraperitoneal injection of gabapentin into the experimental animals immediately after skin incision surgery, This is the result of measuring the number of ultrasonic waves generated in the 22-27 kHz lunar calendar.

 Control group (n = 10), gabapentin (30 mg / kg) administration group (n = 10), * p <0.05 Figure 16 shows the intraperitoneal injection of gabapentin into the experimental animals immediately after skin incision surgery, 3.5 hours, 6 hours, It is the result of measuring the number of times the experimental animal generates ultrasonic waves in the 22-27 kHz lunar calendar after 24 hours.

 Control group (n = 10), gabapentin (30 mg / kg) administration group (n = 10), * p <0.05 Figure 17 shows the intraperitoneal injection of naproxen into the experimental animals immediately after cutaneous surgery, 5 hours after surgery Pain sensitivity was measured by mechanical allodynia evaluation.

 Control group (n = 10), naproxen (30 mg / kg) administration group (n = 10), * p <0.05 Figure 18 is a day after intraperitoneal injection of naproxen in the experimental animals immediately after skin incision surgery (P0D1) The pain sensitivity at the surgical site was measured by mechanical allodynia evaluation.

 Control group (n = 10), naphtoxene (30 mg / kg) administration group (n = 10), * p <0.05 Figure 19 is a laboratory animal 6 hours after intraperitoneal injection of naproxen in the experimental animals immediately after skin incision surgery This is the result of measuring the number of ultrasonic waves generated in the 22-27 kHz lunar calendar.

Control group (n = 10), naproxen (30 mg / kg) administration group (n = 10), * p <0.05 20 is a result of measuring the number of times the experimental animal generates ultrasound in the 22-27 kHz lunar calendar after 3.5 hours, 6 hours, 24 hours after intraperitoneal injection of naproxen in the experimental animals immediately after the skin incision surgery.

 Control group (n = 10), naproxen (30 mg / kg) administration group (n = 10) ᅳ * p <0.05

 FIG. 21 is a result of intraperitoneal injection of gabapentin into an experimental animal for 14 days after nerve branch ligation injury surgery, and then the pain sensitivity was measured by mechanical allodynia evaluation.

 Control group (n = 10), gabapentin (30 mg / kg) administration group (n = 10), *** ρθ.001 Figure 22 shows the baseline of pain threshold value through mechanical allodynia evaluation of pain sensitivity before nerve branch ligation injury surgery Is the result of measuring.

 Control group (n = 10), gabapentin (30 mg / kg) group (n = 10)

 FIG. 23 is a result of intraperitoneal injection of naproxen into an experimental animal for 14 days after nerve branch ligation injury, and then the pain sensitivity was measured by mechanical allodynia evaluation.

 Control group (n = 10), naproxen (30 mg / kg) administration group (η = 10), ** ρθ.01 Figure 24 shows the baseline of the median threshold value of pain sensitivity through mechanical allodynia evaluation before the operation of nerve branch ligation injury It is a result of a measurement.

 Control group (n = 10), naproxen (30 mg / kg) administration group (n = 10)

 25 shows the results of intraperitoneal injection of gabapentin into the experimental animals for 7 days after nerve branch ligation injury surgery, and then the number of times the experimental animals generate ultrasound at 22 kHz 27 kHz lunar zone.

 Control group (n = 10), gabapentin (30 mg / kg) administration group (n = 10), * p <0.05

 FIG. 26 is a result of measuring the number of times the experimental animals generate ultrasound in the 22-27 kHz lunar zone after intraperitoneal injection of naproxen into the experimental animals for 7 days after nerve branch ligation injury surgery.

 Control group (n = 10), naproxen (30 mg / kg) administration group (n = 10), * p <0.05

[Specific contents for implementation of the invention]

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. It will be apparent to those of ordinary skill in the art. Example

Materials and methods

 1. Mate Extract

Natural product extract sample was purchased from Yaksudang (Korea) in Gyeongdong market as a raw material, and extracted by cutting before extraction. 70 vol% alcohol was added 10 times per 100 g of Mate raw material and extracted at 80 ^° C. for 4 hours, and the filtrate and the residue extracts were combined, filtered and concentrated under reduced pressure. All extracts were filtered and concentrated under reduced pressure, lyophilized, and powdered and used in the experiment.

2. Experimental Animal

 Spra ague-Daw ley (SD) rats (200-250 g, Male) were distributed in Samtako Co., Ltd. and used for experiments after being dropped in a breeding box for experimental animals for one week. Animals were kept under conditions of temperature 22 ± rc, humidity 55 ° C 5%, day and night cycle (12 hours day / 12 hours night), and roughness of 300 Lux. Feed and water were freely fed. All animals were managed according to the Guidelines for the Use of Laboratory Animals by KFRI-IACUC (Korea Food Research Institute, Institutional Animal Care and Use Committee).

3. Sample Preparation and Administration

 Body weights were measured and animals were randomly determined after adaptation, and individual identification of experimental animals was carried out using the skin pigmentation method. Mate 70% alcohol extract samples were prepared by dissolving and suspending at a suitable concentration in distilled water. The prepared sample is operated

30 minutes prior to oral administration at a dose of 300 mg / kg / 5 ml.

Secondary distilled water was orally administered 30 minutes prior to surgery.

Gabapentin and naproxen were prepared by dissolving in saline (0.9% NaCl) at an appropriate concentration. The prepared sample was intraperitoneal injection at a dose of 30 mg / kg / 3 ml immediately after surgery. The same amount of saline was injected intraperitoneally immediately after surgery.

4. Pain Animal model _making,

4-1. Skin incision model

 The rat skin incision model was proposed by Brennan (Brennan, 1996). Postoperative pain is generally considered to be a form of acute pain, and rat incision model is considered to be similar to postoperative pain state in humans. SD male rats (200-250 g) under general anesthesia using pentobarbitalol, disinfecting the left foot with 10%, povidone solution, starting at 0.5 cm away from the tip of the heel and extending 1 cm in lengthwise direction. The skin and fascia were cut in 11 times with a surgical knife. The plantar muscle of the incision (foot plantar muscle) was lifted with forceps and separated 1 cm in length. Both ends of the longitudinal direction were carefully lifted up so that the plantar muscles did not fall off, and the incision was gently pressed to hemostatically. The fascia and skin of the hemostatic incision were closed with nylon 4-0 suture and sterilized with 10% povidone solution. After the operation, antibiotic ointment was applied to prevent infection and transferred to a cage for recovery. During the observation period, animals with heat or sutures and animals with edema were excluded from the experiment. 4-2. Spared nerve injury (SNI)

Neural branch ligation injury animal models have been proposed by Decosterd and Wool f (Decosterd & Woolf, 2000). SD male rats (20 g 250 g) were used for general anesthesia using pentobarpital. After the rats under general ^anesthesia, left side view of the skin side of the thigh, and the incision and remove the gluteal daetoeyi (biceps femoris) were exposed to the three terminal branches of the sciatic nerve. The sciatic nerve consists of three strands of nerves: the nasal nerve, the common peroneal never, and the tibial nerve. The double gastrocnemius should remain intact without damage, branching to the right when observed from the left leg reference, the smallest nerve. The total nasal bones and tibias are branched along the muscles in the direction of the legs, and then separated separately, firmly ligated using nylon 4-0 sutures, and then using forceps and scissors. 2-4 mm length was cut off and removed. After the nerve cutting was finished, the muscle layer was sutured, and then the skin incision was resealed and the experimental animals were transferred to a breeding cage for recovery. 5. Evaluation method

 5-1. Von frey filament test

 A method for measuring the degree of mechanical allodynia after animal pain modeling was described by Chaplan et al. (Chaplan et. Al, .1994). The rats were placed in an acrylic triangle placed on a wire mesh test bench with a 2X2 mm mesh size and decanted for at least 15 minutes.When the rat movement became quiet, the pain threshold was measured using a continuous thickness of bone filament (Stoelting, USA). ) Values were evaluated. The filament is placed vertically in contact with the left affected plantar foot and held for 5 to 6 seconds to indicate that the rat exhibits rapid evacuation reactions, or if it immediately flits or licks the plantar foot while releasing hairs. If the stimulation is positive from the bone filament in the center, it is stimulated with a weak filament. If there is no positive reaction, it is stimulated with a strong filament. The minimum stimulus size indicating a positive reaction is used as the threshold, and the upper limit is used when there is no reaction at 15 g or more. 5-2. Ultrasonic vocalization calls

 Rats generate ultrasound when they are in pain, pain, atrophy, or stress, the range of which is reported to be 22-27 KHz (Portfors, 2007). Ultrasound produced by rats when they feel pain using USV measurement system (Sonot rack®, ver 1.5.0, Metris, Netherlands) for 10 minutes to measure the pain level for 3.5 hours, 6 hours and 24 hours after surgery (22—27 KHz) The sound was measured. The rats were placed in an acrylic box capable of measuring ultrasonic waves, stabilized for 15 minutes, and then subjected to ultrasonic measurement experiments for 10 minutes. Experiment result

Confirmation of Mate's Mass Mitigation Effect by Mechanical Allodynia Evaluation Mechanical allodynia evaluation was performed to measure pain sensitivity at the surgical site 5 and 24 hours after the skin incision procedure. As a result, the pain threshold value (g) of the control group was 0.279 ± 0.058 (g) for 5 hours after surgery, whereas the Mate 300 mg / kg group had 1.112 ± 0.375 (g). 05) was confirmed to increase (FIG. 1). In addition, the pain threshold (g) value of the control group (P0D1) was 0.171 ± 0.061 (g) on the postoperative day, whereas the Mate 300 mg / kg group had 1.649 ± 0.69 (g) in the magnitude of the pain-bearing force. p <0.05) (Fig. 2). Confirmation of Mate's Palliative Effect by Ultrasound

 The pain relief effect of mate was measured by quantifying the number of ultrasound lunar zones (22–27 kHz) caused by the animals when they felt pain for 3.5 hours, 6 hours, 24 hours (P0D1) after skin incision. It was confirmed. After 6 hours, the 300 mg / kg group recorded 5.1 ± 1.233 (calls), which was significantly decreased (ρθ.05), compared to 10.4 ± 1.869 (calls). (FIG. 3). On day 1 postoperatively (P0D1), the control group had 14.0 ± 3.296 (calls) in the ultrasound vocal measurements, while the 300 mg / kg group recorded 6.429 ± 1.343 (calls) and significantly decreased (p <0.05). It was confirmed (Fig. 4). Confirmation of Mate's Mass Mitigation Effect by Mechanical Allodynia Evaluation

Mechanical allodynia was evaluated to assess pain sensitivity and oral administration of mate for 14 days after nerve branch ligation injury. As a result, the pain threshold (g) value of the control group was 0.429 ± 0.123 (g), whereas in the Mate 300 mg / kg group, the magnitude of pain-sensing force was significant (ρ <0 · 05). It was confirmed that the increase (Fig. 6). As a result, the pain relief effect of Mate was confirmed in the long-term model as well as the short-term model. Mechanical allodynia evaluation was performed to determine the baseline of pain between the two groups before nerve branch ligation injury. As a result, it was confirmed that the preoperative pain threshold (g) value between the two experimental groups (Fig. 5). Verification of Pain Relief by Positive Control of Gabbapentin by Mechanical Allodynia Evaluation (von frey filament test)

 Five hours after the skin incision, mechanical allodynia was evaluated to measure the pain sensitivity of the surgical site. As a result, the withdrawal threshold (g) value of the control group was 0.141 ± 0.049 (g), whereas the positive control Gabapentin 30 mg / kg group had 7.486 ± 1.308 (g). ρθ.001) was confirmed to increase (Fig. 13). In addition, the pain threshold (g) value of the control group was 0.087 ± 0.027 (g) after 1 day after surgery (P0D1). It was confirmed that the size of significantly increased (p <0.05) (FIG. 14). Ultrasononic Vocalization Calls in the Skin Incision Model to Determine the Pain Relief Effect of Gabbapentin

 3.5 hours, 6 hours, 24 hours (P0D1) after the skin incision surgery, the ultrasonic lunar zone (22-27 kHz) caused by pain in the test animals was measured and the number of times was quantified to reduce the pain of the positive control group Gabapentin. It was confirmed. 3.5 hours after surgery, control group 6.857 ± 1.37 (calls), positive control gabapentin 30 mg / kg group 2JL43 ± 1.37 (calls) (* p <0.05), 6 hours after surgery, control group 19.286 士 5.432 (calls), positive control gabapentin 30 mg / kg group 3.714 ± 1.672 (calls) p <0.05), 24 hours after surgery, control group 24.143 ± 6.967 (calls) positive control group Gabapentin 30 mg / kg group

5.286 ± 1.229 (calls) p <0.05), and recorded a significant decrease (ρ <0 · 05) (Fig. 16). Evaluation of Pain Relief by Positive Control of Naproxen Using Mechanical Allodynia Evaluation in Skin Incision Model

Five hours after the skin incision, mechanical allodynia was evaluated to measure the pain sensitivity of the surgical site. As a result, the pain threshold (g) value of the control group was 0.122 ± 0.025 (g), whereas the positive control naproxen 30 mg / kg group had a significant magnitude of pain-sensing force (4.8 ± 1.528 (g)) (ρ <0 ·). 05) It was confirmed (FIG. 17). In addition, as a result of mechanical allodynia evaluation on the 1st day after surgery, the pain threshold (g) value of the control group was 0.107 ± 0.025 (g), whereas the positive control naproxen 30 mg / kg group had a pain intensity of 3.993 ± 1.698 (g). Was found to increase significantly (p <0.05) (FIG. 18). Confirmation of Pain Relief by Positive Control of Naproxen Using Ultrasound

 3.5 hours, 6 hours, 24 hours (P0D1) after the skin incision, the ultrasonic lunar zone (22-27 kHz) caused by the time of the test animal's pain was measured and the number of times was quantified to determine the pain of the positive control group naproxen. The relaxation effect was confirmed. 6 hours after surgery, control group 19.286 ± 5.432 (calls), positive control group naproxen 30 mg / kg group 5.667 ± 1.382 (calls) (* p <0.05), 24 hours after surgery control group 24.143 ± 6.967 (calls) positive control group naproxen 30 3.833 ± 1.195 (calls) ρ <0 · 05) was recorded in the mg / kg group, and it was confirmed that the decrease was significant (ρ <0.05). After 3.5 hours of surgery, the 22-27 kHz ultrasound recordings of the naproxen 30 mg / kg group decreased compared to the control group (FIG. 20).

Mechanical Allodynia Evaluation in the SNI Model for Efficacy of Gabapentin in the Positive Control Group

Long-term intraperitoneal injection of gabapentin for 14 days after SNI surgery was followed by mechanical allodynia evaluation to determine pain sensitivity. As a result, the gabapentin 30 mg / kg group had a 6.0 0.8 0.882 (g) of pain intensity (ρθ.001) compared to the control group's pain threshold (g) value of 0.429 ± 0.123 (g). It was confirmed that the increase (Fig. 21). Seen _. Experimental results show that gabapentin has a pain-relieving effect in the long-term model as well as the short-term model. Mechanical allodynia evaluation was performed to identify the baseline of pain between the two groups before SNI surgery. As a result, it was confirmed that the preoperative pain threshold (g) ^' value between the two experimental groups (Fig. 22).

Pain Relief in the Positive Control Naproxen by Evaluation of Mechanical Allodynia in the SNI Model Check the effect

 After prolonged intraperitoneal injection of naproxen for 14 days after SNI surgery, mechanical allodynia evaluation was performed to measure pain sensitivity. As a result, the naproxen 30 mg / kg group was 3.88 ± 1.014 (g) in comparison with the control group's pain threshold (g) value of 0.333 ± 0.08 (g) (p <0.01). It was confirmed that the increase (Fig. 23). In this experiment, it was confirmed that naproxen has pain relief effect in the long-term model as well as the short-term model. Mechanical allodynia evaluation was performed to identify the baseline of pain between the two groups before SNI surgery. As a result, it was confirmed that the preoperative pain threshold (g) value between the two experimental groups (Fig. 24).

Ultrasonic Evoked Sounds in SNI Models for Efficacy of Gabapentin Positive Control

 Intraperitoneal injection of gabapentin for 7 days after SNI surgery (P0D7), followed by measurement of the ultrasound lunar zone (22-27 kHz) that causes when the animals feel pain, and quantify the number of times to relieve the pain of gabapentin The effect was confirmed. The control group recorded 19.778 ± 5.482 (calls), Gabapentin 30 mg / kg group 4.571 ± 0.948 (cal ls) (* ρ <0.05), and the 22-27 kHz lunar ultrasound generation was significantly (p <0.05) in the Gabapentin treated group. It was confirmed that the decrease (Fig. 25).

Confirmation of the Mass Relaxation Effect of the Positive Control Naproxen by Measuring Ultrasound in SNI Model

 Intraperitoneal injection of naproxen for 7 days after SNI surgery (P0D7), followed by measurement of the ultrasound lunar zone (22-27 kHz) caused by pain in experimental animals and quantification of the number of times to relieve naproxen pain The effect was confirmed. Control group 17.556 ± 3.783 (calls), naproxen 30 mg / kg group 4.714 ± 0.808 (cal ls) (* p <0.05), 22-27 kHz lunar zone ultrasound generation significantly decreased (p <0.05) in naproxen treated group It was confirmed that (Fig. 26). conclusion

The present inventors (Ilex paraguayensis) extract is a pain animal model skin In both the incision model and the nerve branch ligation injury model, the pain relief effect was confirmed. Especially, the short-term and long-term effects were confirmed. Therefore, using the composition containing the mate extract of the present invention as an active ingredient exhibits the effect of preventing or relieving pain.

 In addition, the present invention is a short-term skin incision model and a long-term model

In the SNI model, it was found that the ultrasonic sound measurement method is an excellent experimental method for the severity and quantification of pain. In the prevention and treatment of pain, it is possible to minimize and overcome errors in individual differences and subjective judgments of the experimenter, which are disadvantages when the evaluation of mechanical allodynia alone is performed as an experimental method for determining the presence and extent of pain. I claim to be able to.

 The pain relief effect of the positive control group Gabapentin, naproxen used in the present invention was clearly demonstrated through the evaluation of the ultrasonic phonation as well as mechanical allodynia. By using the present invention, the pain response can be measured by more objective and scientific methods, and the reliability of the experimental results can be improved. references

 1.Polyphenolic Compounds, Antioxidant Capacity, and Qui none Reductase Activity of an Aqueous Extract of Ardisia compressa in

Compar i son to Mate (Ilex paraguar iensis) and Green (Camellia sinensis) Teas, J. Agric. Food Chem. 52, 3583-3589 (2004).

 2.Recent advances on I lex paraguar iensis research: Minireview, Journal of Ethnopharmacology, 136: 378—384 (2011),

 3. Brennan TJ, Vandermeulen EP, Gebhart GF. Character izat ion of a rat model of incisional pain. Pain 64: 493-501 (1996).

 4. Decosterd I, Wool f CJ. Spared nerve injury: an animal model of persistent peripheral neuropathic pain. Pain 87: 149-58 (2000).

 5. Chap lan SR, Bach FW, Pogrel JW, Chung JM, Yaksh TL. Quantitative assessment of tactile al lodynia in the rat paw. J

Neurosci Methods 53: 55—63 (1994). 6. Portfors CV. Types and functions of ultrasonic vocalizations in laboratory rats and mice. J Am Assoc Lab Anim Sci. 46 (1): 28-34 (2007). The specific parts of the present invention have been described in detail above, and it is apparent to those skilled in the art that these specific technologies are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims

[Patent Claims]

[Claim 1]

 A composition for alleviating, preventing or treating pain comprising an extract of Mate paraguayensis as an active ingredient.

[Claim 2]

 According to claim 1, wherein the composition is a composition, characterized in that the composition for preventing pain.

[Claim 3]

 The method of claim 1, wherein the pain is somatic pain, visceral pain, inflammatory pain, dysfunction pain, idiopathic pain, neuropathic pain, superficial pain, deep pain ( deep pain), itching, migraine and cancer pain.

[Claim 4]

 4. The composition of claim 3, wherein said neuropathic pain is persistent peripheral neuropathic pain.

[Claim 5]

 The composition of claim 1, wherein the composition is a food composition.

[Claim 6]

The composition of claim 1, wherein the composition is a pharmaceutical composition. ^'

[Claim 7]

 A method for alleviating, preventing or treating pain, comprising administering to a subject a composition comprising an extract of Mate paraguayensis as an active ingredient.

[Claim 8]

 8. The method of claim 7, wherein the method is a method for preventing pain:

[Claim 9]

 The method of claim 7, wherein the pain is somatic pain, visceral pain, inflammatory pain, dysfunction pain, idiopathic pain, neuropathic pain, superficial pain, deep pain ( deep pain, itching, migraine and cancer pain.

[Claim 10]

 10. The method of claim 9, wherein the neuropathic pain is persistent peripheral neuropathic pain.

[Claim 111]

 8. The method of claim 7, wherein said composition is a food composition.

[Claim 12]

 8. The method of claim 7, wherein said composition is a pharmaceutical composition.

[Claim 13]

Evaluation of the efficacy of pain relief, prevention or treatment of a sample comprising the following steps: (a) causing pain in an animal other than a human;

 (b) administering a sample to be analyzed to the animal; And

 (c) measuring the 22-27 kHz ultrasound generated from the animal to which the sample is administered, wherein the sample reduces pain when the sample reduces the 22-27 kHz ultrasound generated from the animal; It is determined to have a prophylactic or therapeutic effect.

[Claim 14]

 The method of claim 13, wherein the pain in the pain relief, prevention or treatment efficacy of the sample is somatic pain, visceral pain, inflammatory pain, dysfunction pain, idiopathic pain, neuropathic pain, surface Evaluation method characterized in that it is selected from the group consisting of superficial pain, deep pain, itching, migraine and cancer pain.

[Claim 15]

 14. The method of claim 13, wherein the pain induction is by drug administration, skin incision or nerve ligation.

[Claim 16]

 14. The method of claim 13, wherein the ultrasonic measurement in step (c) measures the frequency or duration of ultrasonication of 22-27 kHz.

[Claim 17]

 14. The method of claim 13, wherein said animal is a rodent animal.

[Claim 18]

 The method of claim 7, wherein the rodent animal is a mouse (Mus musculus) or rat (Rattus norvegicus) evaluation method.