WO2020111541A1 - Composition for preventing or treating pain, containing aconine as active ingredient - Google Patents

Composition for preventing or treating pain, containing aconine as active ingredient Download PDF

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WO2020111541A1
WO2020111541A1 PCT/KR2019/014663 KR2019014663W WO2020111541A1 WO 2020111541 A1 WO2020111541 A1 WO 2020111541A1 KR 2019014663 W KR2019014663 W KR 2019014663W WO 2020111541 A1 WO2020111541 A1 WO 2020111541A1
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pain
aconine
composition
present
sodium channel
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PCT/KR2019/014663
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French (fr)
Korean (ko)
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장익순
최종순
이제호
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한국기초과학지원연구원
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/322—Foods, ingredients or supplements having a functional effect on health having an effect on the health of the nervous system or on mental function

Definitions

  • the present invention relates to a composition for preventing or treating pain, and more particularly, to a composition for preventing, treating or improving pain, comprising aconine or a pharmaceutically acceptable salt thereof as an active ingredient.
  • Pain in humans refers to "unpleasant sensational and emotional experiences due to actual or potential tissue damage or damage.” Is defined. This way, the pain can be avoided from potential dangers, and it will not only protect the damaged body parts until they recover, but also avoid them in the future. Pain usually begins with stimulation to a pain receptor (nociceptor) located in the peripheral nervous system (PNS) or is caused by dysfunction due to damage to the PNS or central nervous system (CNS).
  • PNS peripheral nervous system
  • CNS central nervous system
  • Nav voltage-gated sodium channel
  • the voltage-gated sodium channel (Nav) plays a very important role in pain signaling.
  • Nav is a major biological mediator of electrical signaling because it is the primary mediator of the rapid upstroke of the action potential of multiple excitatory cell types (eg, neurons, skeletal muscle cells, cardiomyocytes).
  • excitatory cell types eg, neurons, skeletal muscle cells, cardiomyocytes.
  • Evidence for the role of these channels in normal physiological phenomena, pathological conditions resulting from mutations in the sodium channel gene, preclinical studies in animal models, and clinical pharmacy of known sodium channel modulators are all Nav in nociception Emphasizes the central role of
  • choo ( ⁇ ) is a tuber root of a genus close-up plant such as aconitume ciliare DC., a perennial plant belonging to the family Asteraceae, which is collected when the leaves dry in autumn and dried in the sun. Choo's efficacy is not only strong, diuretic, and circulatory, but also effective in neuralgia.It is also introduced in Donguibogam, but it can be used as an analgesic, but it is highly toxic and legislation is important. Symptoms such as numbness, nociceptiveness and hypotension appear, and it has been difficult to use it as an analgesic drug, as it is said that it can lead to death after repeating the axis due to respiratory paralysis.
  • the present invention has been devised to solve the above problems, and as a result of various studies conducted by the present inventors, the heated toxic component of aconitine, Aconitine, is heated through water immersion.
  • Aconin can be obtained through hydrolysis, and it is confirmed that the obtained aconin not only ensures safety by removing toxic components, but also exhibits excellent anti-pain effects through rapid absorption even when administered orally in animal models.
  • the present invention was completed based on this.
  • An object of the present invention is to provide a pharmaceutical composition for preventing or treating pain, comprising an aconine represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.
  • Another object of the present invention is to provide a health functional food composition for preventing or improving pain, comprising an aconine represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides a pharmaceutical composition for preventing or treating pain, comprising an aconine represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the aconine may block the sodium channel (sodium channel).
  • the pain may be a sodium channel mediated disease.
  • the disease is visceral pain, acute pain, inflammatory pain, postoperative pain, chronic pain, neuropathic pain, cancer pain, chemotherapy pain, trauma pain, surgical pain, childbirth pain, delivery pain , Persistent pain, peripheral mediated pain, central mediated pain, chronic headache, migraine, sinus headache, tension headache, phantom limb pain, toothache, peripheral nerve damage, HIV-related pain, trigeminal neuralgia, post-herpetic neuralgia , Infringement-acceptable pain, familial erythematosis pain, primary erythematosis pain, familial rectal pain, fibromyalgia, and pain associated with multiple sclerosis (MS).
  • MS multiple sclerosis
  • composition may be administered orally.
  • the present invention provides a health functional food composition for preventing or improving pain, comprising an aconine represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides a method of preventing or treating pain, comprising administering the composition to an individual.
  • the present invention provides a use for preventing or treating pain in the composition.
  • aconine having high safety by removing toxic components was absorbed at a rapid time even when administered orally and exhibited a pain suppressing effect while exhibiting similar effects to commercial products. , It is expected that the safety of side effects will be secured, and may be usefully used for pain prevention, treatment, or improvement.
  • FIG. 1 is for verifying the anti-pain effect of aconine in the visceral pain model
  • FIG. 1a shows abdominal contraction after oral administration of 10 mg/kg aconin in an intraperitoneal administration of acetic acid to induce visceral pain. It is a result of confirming the drug peak time by measuring the degree (abdominal constriction),
  • Figure 1b is a result of evaluating the drug effect according to the administration concentration (5, 10, 25, 30 mg / kg) of aconine.
  • FIG. 2 is for verifying the anti-pain effect of aconin in the acute pain and inflammatory pain models
  • FIG. 2a is a result showing a pain pattern over time after formalin administration
  • FIG. 2b is a phase after the formalin test
  • FIG. 2c is the concentration of administration of aconin in phase II after the formalin test (10, 14, 20 mg/kg).
  • FIG. 3 is for verifying the anti-pain effect of aconin in a postoperative pain model.
  • FIG. 3a is a paw withdrawal threshold (PWT) according to oral administration of aconin after incision of the rat's hind paw to induce pain due to tissue damage. Is a result of confirming the drug peak time
  • FIG. 3B is a result of evaluating the drug effect by measuring PWT according to the administration concentration (10, 20, 30 mg/kg) of aconin compared to a commercial drug (Lyrica). .
  • PWT paw withdrawal threshold
  • the present inventors conducted various studies related to the treatment of pain, and as a result, toxicity was removed from Aconitine of Choo to obtain a safe aconine, and the pain inhibitory activity of the aconine As a result of the verification, the present invention was completed by confirming that it exhibits a significant pain-inhibiting effect with rapid absorption even when administered with low activity compared to an injection.
  • the present invention provides a pharmaceutical composition for preventing or treating pain, comprising an aconine represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the aconine represented by Chemical Formula 1 which is an active ingredient for suppressing pain, is hydrolyzed to heat Aconitine, which is a toxic component of Choo, through a water-repellent cloth. It can be obtained through this, at this time, while undergoing the hydrolysis process, the original toxic components are removed to ensure safety.
  • the pain according to the present invention may be a sodium channel mediated disease, wherein the sodium channel mediated disease is visceral pain, acute pain, inflammatory pain, postoperative pain, chronic pain, neuropathic pain, cancer pain, chemotherapy pain, trauma pain, Surgical pain, childbirth pain, delivery pain, persistent pain, peripheral mediated pain, central mediated pain, chronic headache, migraine, sinus headache, tension headache, phantom limb pain, toothache, peripheral nerve injury, pain associated with HIV , Trigeminal neuralgia, post-herpetic neuralgia, nociceptive pain, familial erythropathy pain, primary erythropathy pain, familial rectal pain, fibromyalgia, pain associated with multiple sclerosis (MS)
  • MS multiple sclerosis
  • Suppression of pain according to the pharmaceutical composition of the present invention is due to the blocking of the nacre channel, wherein the term "sodium channel” used in the present invention is a sodium channel because it is essential for the initiation and propagation of electrical signals in the nervous system. Proper and proper functioning of is essential to the normal functioning of the nerve.
  • abnormal Nav channels play an important role in various diseases such as epilepsy, arrhythmia, myotonia, ataxia, multiple sclerosis and irritable bowel syndrome, incontinence and visceral pain, depression, and pain.
  • 10 types of Nav channels have been reported in humans (Nav1.1 ⁇ 1.9, Nax). Of these, 4 types of Nav1.3, Nav1.7, Nav1.8, and Nav1.9 channels are closely related to the transmission of pain signals. Known to be relevant, it is recognized as an important pain reliever target.
  • prevention refers to all actions that suppress pain or delay the onset of disease by administration of the pharmaceutical composition according to the present invention.
  • treatment refers to all actions in which pain symptoms are improved or beneficially altered by administration of a pharmaceutical composition according to the present invention.
  • aconin was prepared from aconitin, a toxic component of Choo (see Example 1), and after oral administration of aconin to a mouse model to confirm the anti-pain effect of the aconin, acetic acid As a result of intraperitoneal administration to the mouse model to induce visceral pain, it was confirmed that abdominal contraction was suppressed according to administration of aconin (see Example 2).
  • aconin exhibits a similar effect to that of a commercial product while acting at a fast time with anti-analgesic effect with rapid absorption into the body through oral administration, suggesting that it may be applicable as various analgesics in the future. do.
  • the salt is an acid addition salt formed by a pharmaceutically acceptable free acid.
  • Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes It is obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids.
  • These pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulphite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, meth Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenes
  • the acid addition salt according to the present invention is dissolved in a conventional method, for example, a compound represented by the formula (1) in an excess aqueous acid solution, and the salt is water-miscible organic solvent, such as methanol, ethanol, acetone or acetonitrile. It can be prepared by precipitation using. It can also be prepared by evaporating the solvent or excess acid from the mixture and then drying it or suction-filtering the precipitated salt.
  • a conventional method for example, a compound represented by the formula (1) in an excess aqueous acid solution
  • the salt is water-miscible organic solvent, such as methanol, ethanol, acetone or acetonitrile. It can be prepared by precipitation using. It can also be prepared by evaporating the solvent or excess acid from the mixture and then drying it or suction-filtering the precipitated salt.
  • the alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the inexpensive compound salt, and evaporating and drying the filtrate. At this time, it is suitable to manufacture sodium, potassium or calcium salts as metal salts.
  • the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).
  • the pharmaceutical composition according to the present invention includes an aconine represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and may also include a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier is commonly used in formulation, and includes, but is not limited to, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, etc. If necessary, it may further contain other conventional additives such as antioxidants, buffers, if necessary.
  • diluents, dispersants, surfactants, binders, lubricants, and the like can be additionally added to prepare formulations for injection, pills, capsules, granules, or tablets, such as aqueous solutions, suspensions, and emulsions.
  • suitable pharmaceutically acceptable carriers and formulations the formulations described in Remington's literature can be used to formulate according to each component.
  • the pharmaceutical composition of the present invention is not particularly limited in the formulation, but may be formulated as an injection, an inhalant, an external preparation for skin, or an oral intake.
  • the pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenously, subcutaneously, skin, nasal, and airways) according to a desired method, preferably orally, but may be administered orally It can be appropriately selected according to the state of the.
  • the dosage varies depending on the patient's condition and body weight, the degree of disease, the drug form, the administration route and time, but can be appropriately selected by those skilled in the art.
  • composition according to the invention is administered in a pharmaceutically effective amount.
  • a pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of patient's disease, severity, and activity of the drug , Sensitivity to drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-drugs and other factors well known in the medical field.
  • the composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art.
  • the effective amount of the composition according to the present invention may vary depending on the patient's age, sex, and body weight, and is generally administered in an amount of 0.001 to 150 mg per kg of body weight, preferably 0.01 to 100 mg daily or every other day, or 1 It can be administered by dividing it 1 to 3 times a day.
  • the dosage may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, and age, the above dosage does not limit the scope of the present invention in any way.
  • the present invention provides a health functional food composition for pain improvement comprising an aconine represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the term “improvement” refers to any action that at least reduces the severity of parameters associated with the condition being treated, for example pain symptoms.
  • the active ingredient may be added to the food as it is or used with other foods or food ingredients, and may be suitably used according to conventional methods.
  • the mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (for prevention or improvement).
  • the composition of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less with respect to the raw materials, in the manufacture of a food or beverage.
  • the amount may be below the above range.
  • the health functional food composition of the present invention is an essential ingredient in the indicated ratio, and in addition to containing the above-mentioned active ingredient, there is no particular restriction on other ingredients, and it may contain various flavoring agents or natural carbohydrates, etc., as additional ingredients, such as ordinary drinks.
  • natural carbohydrates described above include monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, etc.; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents such as taumatin and stevia extract (for example, rebaudioside A, glycyrrhizine)
  • synthetic flavoring agents sacharin, aspartame, etc.
  • the proportion of natural carbohydrates can be appropriately determined by the choice of those skilled in the art.
  • the health functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid used in carbonated beverages, and the like. These ingredients can be used independently or in combination. The proportions of these additives can also be appropriately selected by those skilled in the art.
  • the present invention provides a method of preventing, controlling or treating pain comprising administering the pharmaceutical composition to an individual.
  • “individual” means a subject in need of a method for preventing, controlling or treating a disease, and more specifically, a human or non-human primate, mouse, rat, dog, cat Means mammals, such as horses and cows.
  • the aconin according to the present invention is a component separated from Choo ( ⁇ ), and when boiling the aquatinin, the poisonous component of Choo, through boiled water, the aconitin is hydrolyzed and has no toxicity.
  • Aconine and benzoylaconine can be obtained.
  • the dose was adjusted based on the body weight (BW) of the mouse and administered orally once 30 minutes before the experiment, At this time, it was confirmed that the concentration of aconin administered orally was 10 mg/kg.
  • Visceral pain is a common form of pain caused by a disease.
  • 0.6 acetic acid was administered intraperitoneally to induce visceral pain, and then performed a torsion test. To measure the degree of pain. More specifically, while maintaining the fasting (fasting) of the ICR mouse (5 weeks old, male) 2 hours before the start of the torsion test, at the same time, it was adapted to the temperature and humidity of the laboratory where the experiment was to be conducted, and the weight of each mouse was measured before the experiment. It was measured.
  • a 0.6% acetic acid solution was prepared while adapting the mouse to the experimental environment and stored in a light-blocked area.
  • aconin was orally administered 0.5, 1 and 2 hours before the acetic acid prepared above was administered.
  • 0.6% acetic acid was intraperitoneally administered (intraperitoneal injection, I.P) at a concentration of 10 ⁇ l/g according to the body weight of the mouse, and then the degree of abdominal contraction (abdominal constriction) was measured.
  • I.P intraperitoneal injection
  • 0 to 5 minutes was excluded from the test results because of the adaptation time, and the writhing score was measured every 5 minutes for 15 minutes.
  • oral administration compared to administration methods such as intravenous injection, intramuscular injection and subcutaneous injection has a low activation rate, but the oral administration of aconin shows the effect of the drug in the body at 0.5 hours. It means fast absorption and effect, and it is expected that aconin can be usefully used as an analgesic in the future.
  • the formalin test is a model for studying acute and chronic hypersensitivity-reactive neurological changes after inducing the activity of peripheral nerve fibers, and is divided into Phase I and II.
  • the Phase I is acute pain, a reaction caused by direct stimulation of the peripheral nerve, and Phase II is considered to be inflammatory pain caused by inflammation caused by damage to the nerve endings and tissues. It is a model.
  • the formalin test was conducted to induce acute pain and inflammatory pain from the above, and more specifically, while maintaining the fasting condition of the mouse (5 weeks old, male) 2 hours before the start of the experiment, at the same time, the temperature of the laboratory where the experiment will proceed. And humidity. Meanwhile, 5% formalin was prepared using 10% Neutral Buffered Formalin while the mice were adapted to the experimental environment. Next, 30 minutes before the start of the experiment, aconin was orally administered to the mice at concentrations of 10, 14 and 20 mg/kg. Thereafter, the 5% formalin prepared above was injected with 20 ⁇ l into the right instep of the mouse using a 50 ⁇ l syringe equipped with a 26 G needle (Hamilton syringe).
  • Phase II pain response was analyzed by separately measuring secondary pain phenomena occurring 20 minutes after formalin administration at 5 minute intervals for 30 minutes. At this time, as shown in Figure 2a, Phase I and II were able to be distinguished through a pattern of pain induction over time after formalin administration.
  • aconin As a result of evaluating the pain reduction effect according to the concentration of aconin by quantifying the pain in Phases I and II with Area Under the Curve (AUC), as shown in FIGS. 2B and 2C, aconin was 14 mg/ When administered over kg, it was confirmed that each pain in Phases I and II was reduced by 50% or more. From the above results, it was found that aconin shows effective effects in acute pain and chronic inflammatory pain.
  • Postoperative pain is caused by incision of skin and tissue during surgery and is an important model for patients because it is involved in well-being after surgery.
  • the post-surgery pain model used a 5-week-old male SD-rat (Spraque Dawley Rat), wherein the 5-week-old male SD-rat was used for 7 days to increase the accuracy of the experiment by minimizing differences between individuals.
  • a Von Frey Monofilament (VFM) test was performed one day before surgery, and rats with scores within range were selected using Dixon's up-down method.
  • rats within the range were selected by using a high VFM value in the non-operated group and a low VFM value in the postoperative pain model.
  • a pain model was created through an incision about 1 cm incision of the left hind paw of the rat.
  • the suitability of the pain model was determined using the same method of calculating the score one day before surgery.
  • drug peak times were sequentially measured for 0.5, 1 and 2 hours.
  • Pain model ED 50 (median effective dose 50, mg/kg) Visceral pain 33.6 Acute pain 14.4 Inflammatory pain 20.2 Post-operative pain 25.1
  • the aconin obtained according to the present invention is not only a natural product but also a stable substance with no toxicity, and exhibits a significant pain-inhibiting effect with rapid absorption even when administered orally with low activity compared to injections. It is expected to be useful as a therapeutic or improvement material.

Abstract

The present invention relates to a composition for preventing, treating or alleviating pain, containing, as an active ingredient, aconine or a pharmaceutically acceptable salt thereof. The inventors of the present invention have ascertained that: a toxic component of natural product-derived aconine, obtained by the hydrolysis of aconitine, which is a toxic component of Aconiti Kusnezoffii Tuber, through heating after passing a water immersion fabric, is removed so that safety is ensured; rapid absorption and pain inhibition effects are exhibited even during oral administration, which has lower activity than injections; and effects similar to those of commercial products are exhibited, and thus it is expected that safety against side effects is ensured so that the present invention is effectively usable in preventing, treating or alleviating pain.

Description

아코닌을 유효성분으로 포함하는 통증 예방 또는 치료용 조성물 Composition for preventing or treating pain comprising aconin as an active ingredient
본 발명은 통증 예방 또는 치료용 조성물에 관한 것으로서, 보다 구체적으로, 아코닌 (aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 통증 예방, 치료 또는 개선용 조성물에 관한 것이다.The present invention relates to a composition for preventing or treating pain, and more particularly, to a composition for preventing, treating or improving pain, comprising aconine or a pharmaceutically acceptable salt thereof as an active ingredient.
IASP (International Association for the Study of pain) 연구에서 인간에서의 통증을 "실제적, 잠재적인 조직 손상 또는 피해로 인한 불쾌한 감각적, 감정적 경험을 말한다." 라고 정의를 내리고 있다. 이렇게 통증은 잠재적 위험으로부터 피할 수 있도록 하며, 손상된 신체 부위가 회복될 때까지 보호할 뿐만 아니라 앞으로 일어날 상황에 대해서 회피할 수 있게 한다. 통증은 주로 PNS (peripheral nervous system)에 위치한 통증수용체 (nociceptor)로의 자극으로 시작되거나 PNS 또는 CNS (central nervous system)의 손상에 따른 기능장애에 의해 비롯된다.In the International Association for the Study of Pain (IASP), pain in humans refers to "unpleasant sensational and emotional experiences due to actual or potential tissue damage or damage." Is defined. This way, the pain can be avoided from potential dangers, and it will not only protect the damaged body parts until they recover, but also avoid them in the future. Pain usually begins with stimulation to a pain receptor (nociceptor) located in the peripheral nervous system (PNS) or is caused by dysfunction due to damage to the PNS or central nervous system (CNS).
현재, 전 세계적으로 질병 치료 및 더 나은 삶의 질을 위해 보건 복지 분야에 많은 시간을 들여 노력하고 있으며, 제약시장은 진통제 (analgesic) 연구에 비중을 점차 크게 두고 있으나, 많은 진통제 (analgesic)들은 대다수가 합성 신약일 뿐만 아니라 내성, 약물 의존성이라는 부작용을 동반한다. 따라서 합성 신약이 아닌 안전한 천연물 생약 발굴이 필요한 실정이다.Currently, we are spending a lot of time in the field of health and welfare for disease treatment and better quality of life worldwide, and the pharmaceutical market is increasingly focusing on analgesic research, but many analgesic are the majority Not only is it a synthetic new drug, it has side effects such as resistance and drug dependence. Therefore, there is a need to discover safe natural products rather than synthetic new drugs.
전압-개폐 나트륨 채널 (Nav)은 통증 신호전달에 있어서 매우 중요한 역할을 한다. Nav는 전기적 신호전달의 주요한 생물학적 매개자인데, 이는 다수의 흥분성 세포 타입 (예를 들면, 뉴런, 골격근세포, 심근세포)들의 활동 전위의 급속 상행각 (upstroke)의 일차적인 매개자이기 때문이다. 정상적인 생리학적 현상에서의 이들 채널의 역할에 대한 증거, 나트륨 채널 유전자의 돌연변이로부터 발생하는 병리학적 상태, 동물 모델에서의 임상전 연구, 및 공지된 나트륨 채널 조절제의 임상 약학은 모두, 통각에 있어서의 Nav의 중심적인 역할을 강조한다.The voltage-gated sodium channel (Nav) plays a very important role in pain signaling. Nav is a major biological mediator of electrical signaling because it is the primary mediator of the rapid upstroke of the action potential of multiple excitatory cell types (eg, neurons, skeletal muscle cells, cardiomyocytes). Evidence for the role of these channels in normal physiological phenomena, pathological conditions resulting from mutations in the sodium channel gene, preclinical studies in animal models, and clinical pharmacy of known sodium channel modulators are all Nav in nociception Emphasizes the central role of
뉴런 신호의 개시 및 전달에 있어서의 Nav의 역할로 인해, Nav 전류를 감소시키는 길항제는 신경 신호전달을 방지하거나 감소시킬 수 있다. 따라서 Nav 채널이 오래전부터 과흥분이 관찰되는 병태에서 통증을 감소시키기 위한 유망한 표적으로서 간주되면서, 나트륨 채널 차단제 (sodium channel blocker)는 새로운 진통제 약물로 많이 이용되어 왔다. 그러나 이러한 약물들은 사람에게 독성, 의존성, 내성이라는 부작용이 문제가 되어 진통제 개발에 있어 큰 장벽에 부딪혔다.Due to the role of Nav in the initiation and transmission of neuronal signals, antagonists that reduce Nav current can prevent or reduce neuronal signaling. Therefore, while the Nav channel has long been regarded as a promising target for reducing pain in a condition where hyperexcitation is observed, sodium channel blocker has been widely used as a new analgesic drug. However, these drugs faced great barriers to the development of painkillers because of the side effects of toxicity, dependence, and resistance to humans.
한편, 초오 (草烏)는 미나리아재비과에 속한 여러해살이풀인 놋젓가락나물 (aconitume ciliare DC.)과 같은 속 근연 식물의 덩이 뿌리를 건조한 것으로 가을에 잎이 말랐을 때 채취하여 햇볕에 말린다. 초오의 효능은 강심작용, 이뇨작용, 혈액순환 작용뿐만 아니라 신경통에도 효과가 있음이 동의보감에서도 소개되어 있어 진통제로 이용될 수 있으나, 독성이 강하여 법제가 중요하며 수치를 잘못하여 중독을 일으키면 메스꺼움, 사지마비, 통각감퇴, 혈압저하 등의 증상이 나타나고, 호흡중추마비로 추축을 반복하다 죽음에 이를 수 있다고 전해짐에 따라 진통제로서의 사용에는 어려움을 겪어 왔다.On the other hand, choo (草烏) is a tuber root of a genus close-up plant such as aconitume ciliare DC., a perennial plant belonging to the family Asteraceae, which is collected when the leaves dry in autumn and dried in the sun. Choo's efficacy is not only strong, diuretic, and circulatory, but also effective in neuralgia.It is also introduced in Donguibogam, but it can be used as an analgesic, but it is highly toxic and legislation is important. Symptoms such as numbness, nociceptiveness and hypotension appear, and it has been difficult to use it as an analgesic drug, as it is said that it can lead to death after repeating the axis due to respiratory paralysis.
따라서, 상기 부작용 문제를 해결하는 천연물 유래 진통제에 대한 연구 개발이 활발하게 진행되고 있으나 (미국출원특허 13/421809), 아직은 부족한 실정이다.Therefore, research and development for painkillers derived from natural products that solve the above-mentioned side-effect problem has been actively conducted (US Patent No. 13/421809), but it is still insufficient.
본 발명은 상기와 같은 문제점을 해결하기 위해 안출된 것으로서, 본 발명자들은 다양한 연구를 진행한 결과, 초오 (草烏)의 독성 성분인 아코니틴 (Aconitine)을 수침포 (水浸泡)를 거쳐 가열하는 가수분해를 통해 아코닌을 수득할 수 있으며, 상기 수득된 아코닌은 독성 성분이 제거되어 안전성이 확보될 뿐만 아니라, 동물 모델에서 경구 투여 시에도 빠른 흡수를 통한 우수한 항 통증 효과를 나타내는 것을 확인하였는바, 이에 기초하여 본 발명을 완성하였다.The present invention has been devised to solve the above problems, and as a result of various studies conducted by the present inventors, the heated toxic component of aconitine, Aconitine, is heated through water immersion. Aconin can be obtained through hydrolysis, and it is confirmed that the obtained aconin not only ensures safety by removing toxic components, but also exhibits excellent anti-pain effects through rapid absorption even when administered orally in animal models. The present invention was completed based on this.
본 발명의 목적은 하기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 통증 예방 또는 치료용 약학적 조성물을 제공하는 것이다.An object of the present invention is to provide a pharmaceutical composition for preventing or treating pain, comprising an aconine represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.
[화학식 1][Formula 1]
Figure PCTKR2019014663-appb-I000001
Figure PCTKR2019014663-appb-I000001
본 발명의 다른 목적은 상기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 통증 예방 또는 개선용 건강기능식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health functional food composition for preventing or improving pain, comprising an aconine represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다. However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.
상기 목적을 달성하기 위하여, 본 발명은 하기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 통증 예방 또는 치료용 약학적 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating pain, comprising an aconine represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.
[화학식 1][Formula 1]
Figure PCTKR2019014663-appb-I000002
Figure PCTKR2019014663-appb-I000002
본 발명의 일 구현예로, 상기 아코닌 (Aconine)은 나트륨 채널 (sodium channel)을 차단할 수 있다.In one embodiment of the present invention, the aconine (Aconine) may block the sodium channel (sodium channel).
본 발명의 다른 구현예로, 상기 통증은 나트륨 채널 매개 질환일 수 있다.In another embodiment of the invention, the pain may be a sodium channel mediated disease.
본 발명의 또 다른 구현예로, 상기 질환은 내장통증, 급성통증, 염증성통증, 수술 후 통증, 만성통증, 신경병성 통증, 암 통증, 화학요법 통증, 외상 통증, 수술 통증, 출산 통증, 분만통, 지속성 통증, 말초 매개 통증, 중추 매개 통증, 만성 두통, 편두통, 부비동성 두통, 긴장 두통, 환상 사지 (phantom limb) 통증, 치통, 말초 신경 손상, HIV와 연관된 통증, 삼차신경통, 대상포진 후 신경통, 침해수용성 통증, 가족성 홍색사지통증, 원발성 홍색사지통증, 가족성 직장 통증, 섬유근육통, 다발성 경화증(MS)과 연관된 통증으로 이루어진 군으로부터 선택되는 어느 하나일 수 있다.In another embodiment of the present invention, the disease is visceral pain, acute pain, inflammatory pain, postoperative pain, chronic pain, neuropathic pain, cancer pain, chemotherapy pain, trauma pain, surgical pain, childbirth pain, delivery pain , Persistent pain, peripheral mediated pain, central mediated pain, chronic headache, migraine, sinus headache, tension headache, phantom limb pain, toothache, peripheral nerve damage, HIV-related pain, trigeminal neuralgia, post-herpetic neuralgia , Infringement-acceptable pain, familial erythematosis pain, primary erythematosis pain, familial rectal pain, fibromyalgia, and pain associated with multiple sclerosis (MS).
본 발명의 또 다른 구현예로, 상기 조성물은 경구 투여될 수 있다.In another embodiment of the invention, the composition may be administered orally.
또한, 본 발명은 상기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 통증 예방 또는 개선용 건강기능식품 조성물을 제공한다.In addition, the present invention provides a health functional food composition for preventing or improving pain, comprising an aconine represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
또한, 본 발명은 상기 조성물을 개체에 투여하는 단계를 포함하는, 통증 예방 또는 치료방법을 제공한다.In addition, the present invention provides a method of preventing or treating pain, comprising administering the composition to an individual.
또한, 본 발명은 상기 조성물의 통증 예방 또는 치료용도를 제공한다.In addition, the present invention provides a use for preventing or treating pain in the composition.
본 발명의 통증 억제 활성성분으로서, 독성 성분이 제거되어 높은 안전성을 갖는 아코닌 (Aconine)은 경구 투여 시에도 빠른 시간에 흡수되어 통증 억제 효능을 나타내면서, 시판제품과 유사한 효과를 나타내는 것을 확인하였는바, 부작용에 대한 안전성이 확보되어 통증 예방, 치료 또는 개선 용도로 유용하게 이용될 수 있을 것으로 기대된다.As a pain suppressing active ingredient of the present invention, it was confirmed that aconine having high safety by removing toxic components was absorbed at a rapid time even when administered orally and exhibited a pain suppressing effect while exhibiting similar effects to commercial products. , It is expected that the safety of side effects will be secured, and may be usefully used for pain prevention, treatment, or improvement.
도 1은 내장통증 모델에서 아코닌의 항통증 효과를 검증하기 위한 것으로, 도 1a는 아세트산을 복강내투여하여 내장통증을 유발시킨 모델에서 10 mg/kg의 아코닌을 경구 투여한 후 복부수축의 정도 (abdominal constriction)를 측정하여 drug peak time을 확인한 결과이고, 도 1b는 아코닌의 투여 농도(5, 10, 25, 30 mg/kg)에 따른 약물 효과를 평가한 결과이다.1 is for verifying the anti-pain effect of aconine in the visceral pain model, and FIG. 1a shows abdominal contraction after oral administration of 10 mg/kg aconin in an intraperitoneal administration of acetic acid to induce visceral pain. It is a result of confirming the drug peak time by measuring the degree (abdominal constriction), Figure 1b is a result of evaluating the drug effect according to the administration concentration (5, 10, 25, 30 mg / kg) of aconine.
도 2는 급성통증 및 염증성통증 모델에서 아코닌의 항통증 효과를 검증하기 위한 것으로, 도 2a는 포르말린 투여 후 시간 경과에 따른 통증 패턴을 보여주는 결과이고, 도 2b는 포르말린 테스트 (Formalin test) 후 Phase Ⅰ에서 아코닌의 투여 농도(10, 14, 20 mg/kg)에 따른 약물 효과를 평가한 결과이며, 도 2c는 포르말린 테스트 (Formalin test) 후 Phase Ⅱ에서 아코닌의 투여 농도(10, 14, 20 mg/kg)에 따른 약물 효과를 평가한 결과이다. FIG. 2 is for verifying the anti-pain effect of aconin in the acute pain and inflammatory pain models, FIG. 2a is a result showing a pain pattern over time after formalin administration, and FIG. 2b is a phase after the formalin test The result of evaluating the drug effect according to the concentration of administration of aconin (10, 14, 20 mg/kg) in Ⅰ, and FIG. 2c is the concentration of administration of aconin in phase Ⅱ after the formalin test (10, 14, 20 mg/kg).
도 3은 수술 후 통증 모델에서 아코닌의 항통증 효과를 검증하기 위한 것으로, 도 3a는 랫트의 뒷발을 절개하여 조직 손상에 의한 통증을 유발시킨 후 아코닌 경구 투여에 따른 PWT (paw withdrawal threshold)를 측정하여 drug peak time을 확인한 결과이고, 도 3b는 시판 약물 (Lyrica)과 비교하여 아코닌의 투여 농도 (10, 20, 30 mg/kg)에 따른 PWT를 측정하여 약물 효과를 평가한 결과이다.FIG. 3 is for verifying the anti-pain effect of aconin in a postoperative pain model. FIG. 3a is a paw withdrawal threshold (PWT) according to oral administration of aconin after incision of the rat's hind paw to induce pain due to tissue damage. Is a result of confirming the drug peak time, and FIG. 3B is a result of evaluating the drug effect by measuring PWT according to the administration concentration (10, 20, 30 mg/kg) of aconin compared to a commercial drug (Lyrica). .
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명자들은 통증 치료와 관련하여 다양한 연구를 진행한 결과, 초오 (草烏)의 아코니틴 (Aconitine)으로부터 독성이 제거되어 안전한 아코닌 (Aconine)을 수득하였으며, 상기 아코닌의 통증 억제 활성을 검증한 결과, 주사제와 비교하여 활성이 낮은 경구 투여 시에도 빠른 흡수와 함께 유의적인 통증 저해 효과를 나타내는 것을 확인함으로써, 본 발명을 완성하였다.The present inventors conducted various studies related to the treatment of pain, and as a result, toxicity was removed from Aconitine of Choo to obtain a safe aconine, and the pain inhibitory activity of the aconine As a result of the verification, the present invention was completed by confirming that it exhibits a significant pain-inhibiting effect with rapid absorption even when administered with low activity compared to an injection.
이에, 본 발명은 하기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 통증 예방 또는 치료용 약학적 조성물을 제공한다. Accordingly, the present invention provides a pharmaceutical composition for preventing or treating pain, comprising an aconine represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.
[화학식 1][Formula 1]
Figure PCTKR2019014663-appb-I000003
Figure PCTKR2019014663-appb-I000003
본 발명에 있어서, 통증 억제 활성성분인 상기 화학식 1로 표시되는 아코닌 (Aconine)은 초오 (草烏)의 독성 성분인 아코니틴 (Aconitine)을 수침포 (水浸泡)를 거쳐 가열하는 가수분해를 통해 수득할 수 있으며, 이때, 상기 가수분해 과정을 거치면서 본래 초오의 독성 성분이 제거되어 안전성이 확보된다.In the present invention, the aconine represented by Chemical Formula 1, which is an active ingredient for suppressing pain, is hydrolyzed to heat Aconitine, which is a toxic component of Choo, through a water-repellent cloth. It can be obtained through this, at this time, while undergoing the hydrolysis process, the original toxic components are removed to ensure safety.
본 발명의 예방, 치료 또는 개선 대상인 "통증"은 통각수용기와 신경섬유로 구성된 구심성 신경로를 통하여 대뇌피질과 가장자리 계통영역과 맞닿은 부위를 자극함으로써 일어나는 통각 및 감각장애를 말하기도 한다. 이는 신체를 보호하기 위한 방어수단으로서 신체의 안이나 밖에서 일어나는 이상을 전달하는 경고반응이라고 할 수 있다. The "pain", which is the object of prevention, treatment, or improvement of the present invention, refers to nociceptive and sensory disorders caused by stimulating a region in contact with the cerebral cortex and the border system region through afferent nerve pathways composed of nociceptors and nerve fibers. It can be said to be a warning response that conveys abnormalities occurring inside or outside the body as a defense means to protect the body.
본 발명에 따른 통증은 나트륨 채널 매개 질환일 수 있으며, 상기 나트륨 채널 매개 질환은 내장통증, 급성통증, 염증성통증, 수술 후 통증, 만성통증, 신경병성 통증, 암 통증, 화학요법 통증, 외상 통증, 수술 통증, 출산 통증, 분만통, 지속성 통증, 말초 매개 통증, 중추 매개 통증, 만성 두통, 편두통, 부비동성 두통, 긴장 두통, 환상 사지 (phantom limb) 통증, 치통, 말초 신경 손상, HIV와 연관된 통증, 삼차신경통, 대상포진 후 신경통, 침해수용성 통증, 가족성 홍색사지통증, 원발성 홍색사지통증, 가족성 직장 통증, 섬유근육통, 다발성 경화증(MS)과 연관된 통증으로 이루어진 군으로부터 선택되는 어느 하나일 수 있으며, 바람직하게는 내장통증 (visceral pain), 급성통증 (acute pain), 염증성통증 (inflammatory pain) 또는 수술 후 통증 (Post-operative pain)일 수 있으나, 이에 제한되는 것은 아니다.The pain according to the present invention may be a sodium channel mediated disease, wherein the sodium channel mediated disease is visceral pain, acute pain, inflammatory pain, postoperative pain, chronic pain, neuropathic pain, cancer pain, chemotherapy pain, trauma pain, Surgical pain, childbirth pain, delivery pain, persistent pain, peripheral mediated pain, central mediated pain, chronic headache, migraine, sinus headache, tension headache, phantom limb pain, toothache, peripheral nerve injury, pain associated with HIV , Trigeminal neuralgia, post-herpetic neuralgia, nociceptive pain, familial erythropathy pain, primary erythropathy pain, familial rectal pain, fibromyalgia, pain associated with multiple sclerosis (MS) There is, preferably, visceral pain, acute pain, inflammatory pain, or post-operative pain, but is not limited thereto.
본 발명의 약학적 조성물에 따른 통증의 억제는 나트름 채널의 차단을 통해 기인하는 것으로서, 이때, 본 발명에서 사용되는 용어 "나트륨 채널"은 신경계에서 전기 신호 개시 및 전파에 있어서 필수적이기 때문에 나트륨 채널의 적절하고 적합한 기능은 신경의 정상적인 기능에 필수적이다. 궁극적으로, 비정상적인 Nav 채널은 간질, 부정맥, 근긴장증, 운동 실조증, 다발성 경화증 및 과민성 대장증후군, 요실금과 내장통, 우울증, 통증과 같은 각종 질환에서 중요한 역할을 한다. 현재 인간에서 10 종의 Nav 채널이 보고되어 있는데 (Nav1.1~1.9, Nax), 이들 중 Nav1.3, Nav1.7, Nav1.8, Nav1.9 4종의 채널은 통증 신호의 전달과 밀접한 관련성이 있다고 알려져 있어 중요한 진통제 타겟으로 인식되고 있다.Suppression of pain according to the pharmaceutical composition of the present invention is due to the blocking of the nacre channel, wherein the term "sodium channel" used in the present invention is a sodium channel because it is essential for the initiation and propagation of electrical signals in the nervous system. Proper and proper functioning of is essential to the normal functioning of the nerve. Ultimately, abnormal Nav channels play an important role in various diseases such as epilepsy, arrhythmia, myotonia, ataxia, multiple sclerosis and irritable bowel syndrome, incontinence and visceral pain, depression, and pain. Currently, 10 types of Nav channels have been reported in humans (Nav1.1~1.9, Nax). Of these, 4 types of Nav1.3, Nav1.7, Nav1.8, and Nav1.9 channels are closely related to the transmission of pain signals. Known to be relevant, it is recognized as an important pain reliever target.
본 발명에서 사용되는 용어, "예방"이란 본 발명에 따른 약학적 조성물의 투여에 의해 통증을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.As used in the present invention, the term "prevention" refers to all actions that suppress pain or delay the onset of disease by administration of the pharmaceutical composition according to the present invention.
본 발명에서 사용되는 용어, "치료"란 본 발명에 따른 약학적 조성물의 투여에 의해 통증 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다.As used in the present invention, the term "treatment" refers to all actions in which pain symptoms are improved or beneficially altered by administration of a pharmaceutical composition according to the present invention.
본 발명에서는 아코닌에 대한 통증 예방 또는 치료 효과를 검증하고자 다양한 통증 모델을 제작하여 상기 아코닌의 경구 투여를 통한 통증 억제 능력을 확인하는 실험을 수행하였다.In the present invention, in order to verify the effect of preventing or treating pain against aconin, various pain models were prepared to perform an experiment to confirm the ability to suppress pain through oral administration of the aconin.
본 발명의 일실시예에서는 초오의 독성 성분인 아코니틴으로부터 아코닌을 제조하였으며 (실시예 1 참조), 상기 아코닌의 항 통증 효과를 확인하고자 마우스 모델에 아코닌을 경구 투여한 뒤, 아세트산을 상기 마우스 모델에 복강내투여하여 내장통증을 유발시킨 결과, 아코닌 투여에 따라 복부수축이 억제되는 것을 확인하였다 (실시예 2참조).In one embodiment of the present invention, aconin was prepared from aconitin, a toxic component of Choo (see Example 1), and after oral administration of aconin to a mouse model to confirm the anti-pain effect of the aconin, acetic acid As a result of intraperitoneal administration to the mouse model to induce visceral pain, it was confirmed that abdominal contraction was suppressed according to administration of aconin (see Example 2).
본 발명의 다른 일실시예에서는 포르말린 테스트를 통해 Phase Ⅰ 및 Ⅱ에서 나타나는 급성통증 및 염증성통증에 대한 아코닌의 항 통증 억제 효과를 검증한 결과, 아코닌의 경구 투여를 통해 Phase Ⅰ 및 Ⅱ에서의 각각의 통증이 50% 이상 감소하는 것을 확인하였고 (실시예 3 참조), 또한, 랫트의 왼쪽 뒷 발바닥 절개를 통해 수술 후 통증 모델을 제작한 후 아코닌을 경구 투여한 결과, 통증을 효과적으로 억제시키는 것을 확인할 수 있었다 (실시예 4 참조).In another embodiment of the present invention, as a result of verifying the anti-pain inhibitory effect of aconin on acute pain and inflammatory pain in Phases I and II through a formalin test, in Phases I and II through oral administration of aconin It was confirmed that each pain was reduced by 50% or more (see Example 3), and also, after a pain model was produced after surgery through the incision of the left hind paw of the rat, oral administration of aconin resulted in effective suppression of pain. It was confirmed (see Example 4).
상기 결과로부터 아코닌은 경구 투여를 통해서도 체내 빠른 흡수와 함께 항 진통 효능이 빠른 시간에 작용하면서, 시판제품과도 유사한 효과를 나타내는 것을 확인할 수 있었는바, 이는 추후 다양한 진통제로서 적용 가능할 수 있음을 시사한다.From the above results, it was confirmed that aconin exhibits a similar effect to that of a commercial product while acting at a fast time with anti-analgesic effect with rapid absorption into the body through oral administration, suggesting that it may be applicable as various analgesics in the future. do.
본 발명에 있어서, 상기 염은 약학적으로 허용 가능한 유리산(free acid)에 의해 형성된 산 부가염이 유용하다. 산 부가염은 염산, 질산, 인산, 황산, 브롬화수소산, 요드화수소산, 아질산 또는 아인산과 같은 무기산류와 지방족 모노 및 디카르복실레이트, 페닐-치환된 알카노에이트, 하이드록시 알카노에이트 및 알칸디오에이트, 방향족 산류, 지방족 및 방향족 설폰산류와 같은 무독성 유기산으로부터 얻는다. 이러한 약학적으로 무독한 염류로는 설페이트, 피로설페이트, 바이설페이트, 설파이트, 바이설파이트, 니트레이트, 포스페이트, 모노하이드로겐 포스페이트, 디하이드로겐 포스페이트, 메타포스페이트, 피로포스페이트 클로라이드, 브로마이드, 아이오다이드, 플루오라이드, 아세테이트, 프로피오네이트, 데카노에이트, 카프릴레이트, 아크릴레이트, 포메이트, 이소부티레이트, 카프레이트, 헵타노에이트, 프로피올레이트, 옥살레이트, 말로네이트, 석시네이트, 수베레이트, 세바케이트, 푸마레이트, 말리에이트, 부틴-1,4-디오에이트, 헥산-1,6-디오에이트, 벤조에이트, 클로로벤조에이트, 메틸벤조에이트, 디니트로 벤조에이트, 하이드록시벤조에이트, 메톡시벤조에이트, 프탈레이트, 테레프탈레이트, 벤젠설포네이트, 톨루엔설포네이트, 클로로벤젠설포네이트, 크실렌설포네이트, 페닐아세테이트, 페닐프로피오네이트, 페닐부티레이트, 시트레이트, 락테이트, β-하이드록시부티레이트, 글리콜레이트, 말레이트, 타트레이트, 메탄설포네이트, 프로판설포네이트, 나프탈렌-1-설포네이트, 나프탈렌-2-설포네이트 또는 만델레이트를 포함한다.In the present invention, the salt is an acid addition salt formed by a pharmaceutically acceptable free acid. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes It is obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids. These pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulphite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, meth Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycol Rate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate or mandelate.
본 발명에 따른 산 부가염은 통상의 방법, 예를 들면, 화학식 1로 표시되는 화합물을 과량의 산 수용액 중에 용해시키고, 이 염을 수혼화성 유기 용매, 예를 들면 메탄올, 에탄올, 아세톤 또는 아세토니트릴을 사용하여 침전시켜서 제조할 수 있다. 또한 이 혼합물에서 용매나 과량의 산을 증발시킨 후 건조시키거나 또는 석출된 염을 흡입 여과시켜 제조할 수도 있다.The acid addition salt according to the present invention is dissolved in a conventional method, for example, a compound represented by the formula (1) in an excess aqueous acid solution, and the salt is water-miscible organic solvent, such as methanol, ethanol, acetone or acetonitrile. It can be prepared by precipitation using. It can also be prepared by evaporating the solvent or excess acid from the mixture and then drying it or suction-filtering the precipitated salt.
또한, 염기를 사용하여 약학적으로 허용 가능한 금속염을 만들 수도 있다. 알칼리 금속 또는 알칼리 토금속 염은 예를 들면, 화합물을 과량의 알칼리 금속 수산화물 또는 알칼리 토금속 수산화물 용액 중에 용해하고, 비용해 화합물 염을 여과하고, 여액을 증발, 건조시켜 얻는다. 이때, 금속염으로는 나트륨, 칼륨 또는 칼슘염을 제조하는 것이 제약상 적합하다. 이에 대응하는 은염은 알칼리 금속 또는 알칼리 토금속 염을 적당한 음염 (예, 질산은)과 반응시켜 얻는다. It is also possible to make pharmaceutically acceptable metal salts using bases. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the inexpensive compound salt, and evaporating and drying the filtrate. At this time, it is suitable to manufacture sodium, potassium or calcium salts as metal salts. The corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).
본 발명에 따른 약학적 조성물은 상기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하며, 또한 약학적으로 허용 가능한 담체를 포함할 수 있다. 상기 약학적으로 허용 가능한 담체는 제제시에 통상적으로 이용되는 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 사이클로덱스트린, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 리포좀 등을 포함하지만 이에 한정되지 않으며, 필요에 따라 항산화제, 완충액 등 다른 통상의 첨가제를 더 포함할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제, 윤활제 등을 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립, 또는 정제로 제제화할 수 있다. 적합한 약학적으로 허용되는 담체 및 제제화에 관해서는 레밍턴의 문헌에 개시되어 있는 방법을 이용하여 각 성분에 따라 바람직하게 제제화할 수 있다. 본 발명의 약학적 조성물은 제형에 특별한 제한은 없으나 주사제, 흡입제, 피부 외용제, 또는 경구 섭취제 등으로 제제화할 수 있다. The pharmaceutical composition according to the present invention includes an aconine represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and may also include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is commonly used in formulation, and includes, but is not limited to, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, etc. If necessary, it may further contain other conventional additives such as antioxidants, buffers, if necessary. In addition, diluents, dispersants, surfactants, binders, lubricants, and the like can be additionally added to prepare formulations for injection, pills, capsules, granules, or tablets, such as aqueous solutions, suspensions, and emulsions. Regarding suitable pharmaceutically acceptable carriers and formulations, the formulations described in Remington's literature can be used to formulate according to each component. The pharmaceutical composition of the present invention is not particularly limited in the formulation, but may be formulated as an injection, an inhalant, an external preparation for skin, or an oral intake.
본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구투여 (예를 들어, 정맥 내, 피하, 피부, 비강, 기도에 적용)할 수 있으며, 바람직하게는 경구 투여할 수 있으나, 환자의 상태에 따라 적절하게 선택될 수 있다. 또한, 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 시간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenously, subcutaneously, skin, nasal, and airways) according to a desired method, preferably orally, but may be administered orally It can be appropriately selected according to the state of the. In addition, the dosage varies depending on the patient's condition and body weight, the degree of disease, the drug form, the administration route and time, but can be appropriately selected by those skilled in the art.
본 발명에 따른 조성물은 약학적으로 유효한 양으로 투여한다. 본 발명에 있어서, "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명에 따른 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The composition according to the invention is administered in a pharmaceutically effective amount. In the present invention, "a pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of patient's disease, severity, and activity of the drug , Sensitivity to drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-drugs and other factors well known in the medical field. The composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art.
구체적으로, 본 발명에 따른 조성물의 유효량은 환자의 나이, 성별, 체중에 따라 달라질 수 있으며, 일반적으로는 체중 1 kg 당 0.001 내지 150 mg, 바람직하게는 0.01 내지 100 mg을 매일 또는 격일 투여하거나 1 일 1 내지 3 회로 나누어 투여할 수 있다. 그러나 투여 경로, 비만의 중증도, 성별, 체중, 연령 등에 따라서 증감될 수 있으므로 상기 투여량이 어떠한 방법으로도 본 발명의 범위를 한정하는 것은 아니다.Specifically, the effective amount of the composition according to the present invention may vary depending on the patient's age, sex, and body weight, and is generally administered in an amount of 0.001 to 150 mg per kg of body weight, preferably 0.01 to 100 mg daily or every other day, or 1 It can be administered by dividing it 1 to 3 times a day. However, since the dosage may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, and age, the above dosage does not limit the scope of the present invention in any way.
본 발명의 다른 양태로서, 본 발명은 상기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 통증 개선용 건강기능식품 조성물을 제공한다. As another aspect of the present invention, the present invention provides a health functional food composition for pain improvement comprising an aconine represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명에서 사용되는 용어, "개선"이란, 치료되는 상태와 관련된 파라미터, 예를 들면 통증 증상의 정도를 적어도 감소시키는 모든 행위를 의미한다.As used herein, the term "improvement" refers to any action that at least reduces the severity of parameters associated with the condition being treated, for example pain symptoms.
본 발명의 건강기능식품 조성물에서 유효성분을 식품에 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합량은 그의 사용 목적 (예방 또는 개선용)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조 시에 본 발명의 조성물은 원료에 대하여 15 중량% 이하, 바람직하게는 10 중량% 이하의 양으로 첨가된다. 그러나 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있다.In the health functional food composition of the present invention, the active ingredient may be added to the food as it is or used with other foods or food ingredients, and may be suitably used according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (for prevention or improvement). Generally, the composition of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less with respect to the raw materials, in the manufacture of a food or beverage. However, in the case of long-term intake for health and hygiene purposes or for health control purposes, the amount may be below the above range.
본 발명의 건강기능식품 조성물은 지시된 비율로 필수 성분으로서 상기 유효성분을 함유하는 것 외에 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토오스, 수크로오스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 당업자의 선택에 의해 적절하게 결정될 수 있다.The health functional food composition of the present invention is an essential ingredient in the indicated ratio, and in addition to containing the above-mentioned active ingredient, there is no particular restriction on other ingredients, and it may contain various flavoring agents or natural carbohydrates, etc., as additional ingredients, such as ordinary drinks. have. Examples of the natural carbohydrates described above include monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, etc.; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (such as taumatin and stevia extract (for example, rebaudioside A, glycyrrhizine)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates can be appropriately determined by the choice of those skilled in the art.
상기 외에 본 발명의 건강기능식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율 또한 당업자에 의해 적절히 선택될 수 있다.In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid used in carbonated beverages, and the like. These ingredients can be used independently or in combination. The proportions of these additives can also be appropriately selected by those skilled in the art.
한편, 본 발명의 또 다른 양태로서, 본 발명은 상기 약학적 조성물을 개체에 투여하는 단계를 포함하는 통증 예방, 조절 또는 치료방법을 제공한다.On the other hand, as another aspect of the present invention, the present invention provides a method of preventing, controlling or treating pain comprising administering the pharmaceutical composition to an individual.
본 발명에서 "개체"란 질병의 예방, 조절 또는 치료방법을 필요로 하는 대상을 의미하고, 보다 구체적으로는, 인간 또는 비-인간인 영장류, 생쥐(mouse), 쥐(rat), 개, 고양이, 말 및 소 등의 포유류를 의미한다.In the present invention, "individual" means a subject in need of a method for preventing, controlling or treating a disease, and more specifically, a human or non-human primate, mouse, rat, dog, cat Means mammals, such as horses and cows.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시한다. 그러나 하기의 실시예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 하기 실시예에 의해 본 발명의 내용이 한정되는 것은 아니다.Hereinafter, preferred embodiments are provided to help understanding of the present invention. However, the following examples are only provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.
[실시예][Example]
실시예Example 1. 실험준비 및 실험방법 1. Experiment preparation and experiment method
1-1. 1-1. 아코닌Aconin ( ( AconineAconine )의 제조 및 투여) Preparation and Administration
본 발명에 따른 아코닌은 초오 (草烏)로부터 분리된 성분으로, 초오의 독성 성분인 아코니틴 (Aconitine)을 수침포 (水浸泡)를 거쳐 끓이면 상기 아코니틴이 가수분해되어 독성이 없는 아코닌 (Aconine) 및 벤조일아코닌 (benzoylaconine)을 수득할 수 있다. 상기 방법으로 수득된 아코닌을 30 % PEG 400 (polyethylene glycol 400)에 용해시킨 후, 마우스의 체중 (Body weight; BW)을 기준으로 투여량을 조절하여 실험 30 분 전 1 회 경구로 투여하였으며, 이 때 경구로 투여된 아코닌의 농도는 10 mg/kg임을 확인하였다. The aconin according to the present invention is a component separated from Choo (草烏), and when boiling the aquatinin, the poisonous component of Choo, through boiled water, the aconitin is hydrolyzed and has no toxicity. Aconine and benzoylaconine can be obtained. After dissolving the aconin obtained by the above method in 30% PEG 400 (polyethylene glycol 400), the dose was adjusted based on the body weight (BW) of the mouse and administered orally once 30 minutes before the experiment, At this time, it was confirmed that the concentration of aconin administered orally was 10 mg/kg.
1-2. 통계분석1-2. Statistical analysis
투여 농도에 따라 분류된 각 시험군의 측정값은 평균과 표준 오차 평균을 시험결과로 사용하였으며, 각 군의 차이는 One-Way ANOVA로 통계학적 분석을 실시하여 유의한 결과가 보이면 post-hoc 분석으로 Tukey Test를 실시하여 각 군 간의 유의성을 평가하였다. For each test group classified according to the administration concentration, the average and standard error average were used as the test results, and the difference between each group was analyzed by one-way ANOVA and statistical analysis was performed. The Tukey Test was conducted to evaluate the significance of each group.
실시예 2. 아코닌 처리에 따른 내장통증 (visceral pain) 저해 효과 확인Example 2. Confirmation of the effect of inhibiting visceral pain due to aconin treatment
본 실시예에서는 아코닌이 통증 감소 효과를 검증하기 위해, 먼저 내장통증 모델에서 실험을 실시하였다. 내장통증 (visceral pain)은 질병에 의해 발생하는 흔한 형태의 통증으로서, 본 실시예에서는 내장통증을 유도하기 위하여 0.6 아세트산 (acetic acid)를 복강내 투여하였으며, 이후 비틀림 테스트 (Writhing test)를 수행하여 통증의 정도를 측정하고자 하였다. 보다 구체적으로, 상기 비틀림 테스트 시작 2 시간 전 ICR mouse (5 주령, 수컷)를 공복상태 (fasting)를 유지시키면서 이와 동시에 실험이 진행될 실험실의 온도 및 습도에 적응시켰으며, 실험 전 각 마우스의 체중을 측정하였다. 한편, 마우스를 실험환경에 적응시키는 동안 0.6 % 아세트산 (acetic acid) 용액을 제조하였고 빛이 차단된 곳에 보관하였다. 다음으로, 아코닌의 항 통증 효과가 나타나는 peak time을 확인하기 위해서, 상기 제조한 아세트산을 투여하기 0.5, 1 및 2 시간 전에 아코닌을 경구 투여하였다. 이후, 0.6 %의 아세트산을 마우스의 체중에 따라 10 ㎕/g의 농도로 복강내투여 (intraperitoneal injection, I.P)한 후, 복부수축의 정도 (abdominal constriction)를 측정하였다. 이 때, 아세트산 투여 후, 0 내지 5 분은 순응 시간 (adaptation time)이기 때문에 테스트 결과에서 제외시켰으며, 15 분 동안 5 분 간격으로 writhing score를 측정하였다.In this embodiment, in order to verify the pain reduction effect of aconine, an experiment was first conducted in the visceral pain model. Visceral pain is a common form of pain caused by a disease. In this example, 0.6 acetic acid was administered intraperitoneally to induce visceral pain, and then performed a torsion test. To measure the degree of pain. More specifically, while maintaining the fasting (fasting) of the ICR mouse (5 weeks old, male) 2 hours before the start of the torsion test, at the same time, it was adapted to the temperature and humidity of the laboratory where the experiment was to be conducted, and the weight of each mouse was measured before the experiment. It was measured. Meanwhile, a 0.6% acetic acid solution was prepared while adapting the mouse to the experimental environment and stored in a light-blocked area. Next, in order to confirm the peak time of the anti-pain effect of aconin, aconin was orally administered 0.5, 1 and 2 hours before the acetic acid prepared above was administered. Thereafter, 0.6% acetic acid was intraperitoneally administered (intraperitoneal injection, I.P) at a concentration of 10 μl/g according to the body weight of the mouse, and then the degree of abdominal contraction (abdominal constriction) was measured. At this time, after administration of acetic acid, 0 to 5 minutes was excluded from the test results because of the adaptation time, and the writhing score was measured every 5 minutes for 15 minutes.
그 결과, 도 1a에 나타낸 바와 같이, 아코닌을 10 mg/kg의 농도를 투여한 경우, 0.5 시간에서 복부수축의 정도 (abdominal constriction)가 1 및 2 시간 보다 감소하는 것을 확인할 수 있었고, 이는 아코닌 투여 시 약물효과가 0.5 시간에서 가장 좋은 것을 보여주는 결과이다. 또한, 통증을 AUC (Area Under the Curve)로 정량화하여 아코닌의 투여 농도에 따른 내장 통증 감소 효과를 평가한 결과, 도 1b에 나타낸 바와 같이, 아코닌을 25 mg/kg 이상으로 투여한 경우 내장통증이 유의하게 감소된 것을 확인하였다. As a result, as shown in Figure 1a, when the concentration of 10 mg / kg aconine, it was confirmed that the degree of abdominal contraction (abdominal constriction) is reduced than 1 and 2 hours at 0.5 hours, this The result shows that the drug effect is best at 0.5 hours when administered with nin. In addition, the pain was quantified by AUC (Area Under the Curve) to evaluate the effect of reducing visceral pain according to the concentration of administration of aconin, and as shown in FIG. 1B, when aconin was administered at 25 mg/kg or more It was confirmed that the pain was significantly reduced.
상기 결과로부터 정맥주사, 근육주사 및 피하주사 등과 같은 투여방법에 비해 경구 투여는 활성화 속도가 낮음에도 불구하고, 아코닌의 경구 투여 시 체내에서 0.5 시간에 약물의 효과가 나타난다는 것은 아코닌의 매우 빠른 흡수율과 효과를 의미하는 것으로, 향후 아코닌을 진통제로서 유용하게 사용될 수 있을 것으로 기대된다.From the above results, oral administration compared to administration methods such as intravenous injection, intramuscular injection and subcutaneous injection has a low activation rate, but the oral administration of aconin shows the effect of the drug in the body at 0.5 hours. It means fast absorption and effect, and it is expected that aconin can be usefully used as an analgesic in the future.
실시예Example 3. 3. 아코닌Aconin 처리에 따른 급성통증 (acute pain) 및 Treatment of acute pain and 염증성통증Inflammatory pain (inflammatory pain) 저해 효과 확인 (inflammatory pain) check the inhibitory effect
상기 실시예 2의 결과에 더하여, 아코닌에 의한 급성통증 및 염증성통증 감소 효과를 알아보기 위해 포르말린 테스트(Formalin test)를 실시하고자 하였다. 상기 포르말린 테스트는 말초신경 섬유의 활성을 유도한 후, 급성 및 만성 과민 반응성 신경 변화를 연구하기 위한 모델로서, Phase Ⅰ 및 Ⅱ로 나누어진다. 상기 Phase Ⅰ은 급성통증 (acute pain)으로 말초 신경의 직접적인 자극에 의해 일어나는 반응이고, Phase Ⅱ는 신경말단과 조직의 손상에 의한 염증 발생에 의해 유발되는 염증성통증 (inflammatory pain)으로 간주될 수 있는 모델이다. In addition to the results of Example 2, to investigate the effect of reducing acute pain and inflammatory pain caused by aconine, a formalin test was conducted. The formalin test is a model for studying acute and chronic hypersensitivity-reactive neurological changes after inducing the activity of peripheral nerve fibers, and is divided into Phase I and II. The Phase I is acute pain, a reaction caused by direct stimulation of the peripheral nerve, and Phase II is considered to be inflammatory pain caused by inflammation caused by damage to the nerve endings and tissues. It is a model.
이에, 상기로부터 급성통증 및 염증성통증을 유도하기 위해서 포르말린 테스트를 진행하였으며, 보다 구체적으로, 실험 시작 2 시간 전 mouse (5 주령, 수컷)를 공복상태를 유지시키면서, 이와 동시에 실험이 진행될 실험실의 온도 및 습도에 적응시켰다. 한편, 마우스를 실험환경에 적응시키는 동안 10 % 중성 완충 포르말린(Neutral Buffered Formalin)을 이용하여 5 % 포르말린을 제조하였다. 다음으로, 실험 시작 30 분 전에 아코닌을 10, 14 및 20 mg/kg의 농도로 상기 마우스에 경구 투여하였다. 이 후, 상기에서 제조한 5 % 포르말린을 26 G 바늘이 장착된 50 ㎕ 주사기 (Hamilton syringe)를 이용하여 마우스의 오른쪽 발등에 20 ㎕를 주사하였으며, 주사가 끝남과 동시에 5 분 간격으로 15 분 동안 licking time을 초 단위로 측정하였다. 또한, Phase Ⅱ 통증 반응은 포르말린 투여 20분 후에 나타나는 2 차적인 통증 현상을 30 분 동안 5 분 간격으로 별도로 측정하여 분석하였다. 이 때, 도 2a에 나타낸 바와 같이, 포르말린 투여 후 시간 경과에 따른 통증 유발의 패턴을 통해 Phase Ⅰ 및 Ⅱ를 구분할 수 있었다. Thus, the formalin test was conducted to induce acute pain and inflammatory pain from the above, and more specifically, while maintaining the fasting condition of the mouse (5 weeks old, male) 2 hours before the start of the experiment, at the same time, the temperature of the laboratory where the experiment will proceed. And humidity. Meanwhile, 5% formalin was prepared using 10% Neutral Buffered Formalin while the mice were adapted to the experimental environment. Next, 30 minutes before the start of the experiment, aconin was orally administered to the mice at concentrations of 10, 14 and 20 mg/kg. Thereafter, the 5% formalin prepared above was injected with 20 μl into the right instep of the mouse using a 50 μl syringe equipped with a 26 G needle (Hamilton syringe). The licking time was measured in seconds. In addition, Phase II pain response was analyzed by separately measuring secondary pain phenomena occurring 20 minutes after formalin administration at 5 minute intervals for 30 minutes. At this time, as shown in Figure 2a, Phase I and II were able to be distinguished through a pattern of pain induction over time after formalin administration.
상기 Phase Ⅰ 및 Ⅱ에서의 통증을 AUC (Area Under the Curve)로 정량화하여 아코닌의 투여 농도에 따른 통증 감소 효과를 평가한 결과, 도 2b 및 도 2c에 나타낸 바와 같이, 아코닌을 14 mg/kg 이상 투여한 경우 Phase Ⅰ 및 Ⅱ에서의 각각의 통증이 50 % 이상 감소하는 것을 확인하였다. 상기 결과로부터 아코닌이 급성통증 및 만성 염증성통증에서도 유효한 효과를 나타내는 것을 알 수 있었다.As a result of evaluating the pain reduction effect according to the concentration of aconin by quantifying the pain in Phases I and II with Area Under the Curve (AUC), as shown in FIGS. 2B and 2C, aconin was 14 mg/ When administered over kg, it was confirmed that each pain in Phases I and II was reduced by 50% or more. From the above results, it was found that aconin shows effective effects in acute pain and chronic inflammatory pain.
실시예Example 4. 4. 아코닌Aconin 처리에 따른 수술 후 통증 (Post-operative pain) 저해 효과 확인 Confirmation of post-operative pain inhibition effect according to treatment
마지막으로, 수술 후 통증 모델에서 아코닌에 의한 통증 감소효과를 검증하고자 하였다. 수술 후 통증은 수술 시 피부와 조직을 절개함으로써 유발되는 것으로 환자들에게 있어 수술 후 well-being에 관여하기 때문에 중요한 모델이다.Lastly, it was intended to verify the pain reduction effect by aconine in the postoperative pain model. Postoperative pain is caused by incision of skin and tissue during surgery and is an important model for patients because it is involved in well-being after surgery.
보다 구체적으로, 상기 수술 후 통증 모델은 5 주령의 수컷 SD-랫트 (Spraque Dawley Rat)를 이용하였으며, 이때, 개체 간 차이를 최소화하여 실험의 정확성을 높이기 위해 5 주령 수컷 SD-랫트를 7 일 동안 충분히 순응시킨 후, 수술 하루 전 Von Frey Monofilament (VFM) 테스트를 수행하고 딕슨의 업-다운 방법 (Dixon's up-down method)을 이용해 범위 내 score를 가진 랫트를 선별하였다. 즉, 수술을 하지 않은 그룹에서는 VFM 값이 높고, 수술 후 통증 모델에서는 낮은 VFM 값에서도 반응하는 것을 이용하여 범위 내 랫트를 선별하였다. 이 후, 상기 랫트의 왼쪽 뒷 발바닥을 1 cm 가량 절개하는 수술을 통해 통증 모델을 만들었으며, 이 때, 상기 통증 모델 적합성은 수술 하루 전 score를 산출하는 동일한 방법을 이용하여 판단하였다. 다음으로, 통증이 형성된 랫트에게 30 % PEG 400에 용해시킨 아코닌을 30 mg/kg의 농도로 경구 투여한 후, 0.5, 1 및 2 시간 동안 순차적으로 drug peak time을 측정하였다. More specifically, the post-surgery pain model used a 5-week-old male SD-rat (Spraque Dawley Rat), wherein the 5-week-old male SD-rat was used for 7 days to increase the accuracy of the experiment by minimizing differences between individuals. After sufficiently acclimatized, a Von Frey Monofilament (VFM) test was performed one day before surgery, and rats with scores within range were selected using Dixon's up-down method. In other words, rats within the range were selected by using a high VFM value in the non-operated group and a low VFM value in the postoperative pain model. Subsequently, a pain model was created through an incision about 1 cm incision of the left hind paw of the rat. At this time, the suitability of the pain model was determined using the same method of calculating the score one day before surgery. Next, after oral administration of aconin dissolved in 30% PEG 400 to a pain-forming rat at a concentration of 30 mg/kg, drug peak times were sequentially measured for 0.5, 1 and 2 hours.
그 결과, 도 3a에 나타낸 바와 같이, 30 mg/kg의 약물을 투여한 경우, PWT (paw withdrawal threshold)가 0.5 시간에서 가장 높게 나타나 수술하지 않은 일반 그룹 (Normal)과 유사한 값을 나타내는 것을 확인할 수 있었다.As a result, as shown in Figure 3a, when administered with a drug of 30 mg / kg, it was confirmed that the PWT (paw withdrawal threshold) was highest at 0.5 hours, indicating a value similar to the normal group (Normal) without surgery. there was.
또한, 아코닌 농도에 따른 효과를 관찰한 결과, 도 3b에 나타낸 바와 같이, 10 및 20 mg/kg 농도로 투여한 경우에는 효과가 미비하게 나타났으나, 30 mg/kg 농도로 투여한 경우 통증 억제효과가 현저히 높게 나타났다. 아울러, 시중에서 판매되고 있는 Lyrica 약물군 (Pfizer)을 30 mg/kg 농도로 투여한 결과와 통증 감소 효과가 유사하게 나타난 것을 확인하였다. 상기 결과로부터 아코닌의 항 진통 효능은 빠른 시간에 작용하면서, 시판제품과 유사한 효과를 나타냄을 알 수 있었다.In addition, as a result of observing the effect according to the concentration of aconin, as shown in Fig. 3b, when administered at concentrations of 10 and 20 mg/kg, the effect was insignificant, but pain when administered at a concentration of 30 mg/kg The inhibitory effect was remarkably high. In addition, it was confirmed that the result of administration of a commercially available Lyrica drug group (Pfizer) at a concentration of 30 mg/kg was similar to the pain reduction effect. From the above results, it was found that the anti-analgesic effect of aconin acts at a rapid time and exhibits similar effects to commercial products.
실시예 5. 다양한 통증 모델에서의 아코닌의 ED50 값 확인Example 5. Confirmation of ED50 values of aconin in various pain models
상기 실시예 2 내지 4에 개시된 통증 모델에서 약물 효과를 나타내는 아코닌의 유효 농도를 확인하기 위하여, 아코닌의 농도를 달리하여 약물의 효과를 평가함으로써 ED 50 (Effective dose 50)을 구하였으며, 그 결과를 하기 표 1에 나타내었다.In order to confirm the effective concentration of aconin exhibiting drug effect in the pain models disclosed in Examples 2 to 4, ED 50 (Effective dose 50) was obtained by evaluating the effect of the drug by varying the concentration of aconin. The results are shown in Table 1 below.
Pain modelPain model ED 50ED 50 (median effective dose 50, mg/kg)(median effective dose 50, mg/kg)
내장통증 (visceral pain)Visceral pain 33.633.6
급성통증 (acute pain)Acute pain 14.414.4
염증성통증 (inflammatory pain)Inflammatory pain 20.220.2
수술 후 통증 (Post-operative pain)Post-operative pain 25.125.1
전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.The above description of the present invention is for illustration only, and a person having ordinary knowledge in the technical field to which the present invention pertains can understand that it can be easily modified to other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.
본 발명에 따라 수득된 아코닌은 천연물일 뿐만 아니라 독성이 없는 안정적인 물질이고, 주사제와 비교하여 활성이 낮은 경구 투여 시에도 빠른 흡수와 함께 유의적인 통증 저해 효과를 나타내는 것을 확인하였는바, 통증 예방, 치료 또는 개선용 물질로 유용하게 이용될 수 있을 것으로 기대된다.It was confirmed that the aconin obtained according to the present invention is not only a natural product but also a stable substance with no toxicity, and exhibits a significant pain-inhibiting effect with rapid absorption even when administered orally with low activity compared to injections. It is expected to be useful as a therapeutic or improvement material.

Claims (9)

  1. 하기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 통증 예방 또는 치료용 약학적 조성물.Aconine represented by Formula 1 (Aconine) or a pharmaceutical composition for the prevention or treatment of pain comprising a pharmaceutically acceptable salt thereof as an active ingredient.
    [화학식 1][Formula 1]
    Figure PCTKR2019014663-appb-I000004
    Figure PCTKR2019014663-appb-I000004
  2. 제1항에 있어서, According to claim 1,
    상기 아코닌 (Aconine)은 나트륨 채널 (sodium channel)을 차단하는 것을 특징으로 하는, 조성물.The aconine (Aconine), characterized in that to block the sodium channel (sodium channel), the composition.
  3. 제1항에 있어서, According to claim 1,
    상기 통증은 나트륨 채널 매개 질환인 것을 특징으로 하는, 조성물.The pain is characterized in that the sodium channel mediated disease, composition.
  4. 제3항에 있어서, According to claim 3,
    상기 질환은 내장통증, 급성통증, 염증성통증, 수술 후 통증, 만성통증, 신경병성 통증, 암 통증, 화학요법 통증, 외상 통증, 수술 통증, 출산 통증, 분만통, 지속성 통증, 말초 매개 통증, 중추 매개 통증, 만성 두통, 편두통, 부비동성 두통, 긴장 두통, 환상 사지 (phantom limb) 통증, 치통, 말초 신경 손상, HIV와 연관된 통증, 삼차신경통, 대상포진 후 신경통, 침해수용성 통증, 가족성 홍색사지통증, 원발성 홍색사지통증, 가족성 직장 통증, 섬유근육통, 다발성 경화증(MS)과 연관된 통증으로 이루어진 군으로부터 선택되는 어느 하나인 것을 특징으로 하는, 조성물.These diseases include visceral pain, acute pain, inflammatory pain, postoperative pain, chronic pain, neuropathic pain, cancer pain, chemotherapy pain, trauma pain, surgical pain, childbirth pain, labor pain, persistent pain, peripheral mediated pain, central Mediated pain, chronic headache, migraine, sinus headache, tension headache, phantom limb pain, toothache, peripheral nerve damage, HIV-related pain, trigeminal neuralgia, postherpetic neuralgia, incidence pain, familial erythema The composition characterized in that it is any one selected from the group consisting of pain, primary erythematosis pain, familial rectal pain, fibromyalgia, and pain associated with multiple sclerosis (MS).
  5. 제1항에 있어서, According to claim 1,
    상기 조성물은 경구 투여되는 것을 특징으로 하는, 조성물.The composition is characterized in that it is administered orally, composition.
  6. 하기 화학식 1로 표시되는 아코닌 (Aconine) 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는, 통증 예방 또는 개선용 건강기능식품 조성물.Aconine represented by the following formula (Aconine) or a pharmaceutically acceptable salt thereof as an active ingredient, pain preventing or improving health functional food composition.
    [화학식 1][Formula 1]
    Figure PCTKR2019014663-appb-I000005
    Figure PCTKR2019014663-appb-I000005
  7. 제6항에 있어서, The method of claim 6,
    상기 아코닌 (Aconine)은 나트륨 채널 (sodium channel)을 차단하는 것을 특징으로 하는, 조성물.The aconine (Aconine), characterized in that to block the sodium channel (sodium channel), the composition.
  8. 제6항에 있어서, The method of claim 6,
    상기 통증은 나트륨 채널 매개 질환인 것을 특징으로 하는, 조성물.The pain is characterized in that the sodium channel mediated disease, the composition.
  9. 제8항에 있어서, The method of claim 8,
    상기 질환은 내장통증, 급성통증, 염증성통증, 수술 후 통증, 만성통증, 신경병성 통증, 암 통증, 화학요법 통증, 외상 통증, 수술 통증, 출산 통증, 분만통, 지속성 통증, 말초 매개 통증, 중추 매개 통증, 만성 두통, 편두통, 부비동성 두통, 긴장 두통, 환상 사지 (phantom limb) 통증, 치통, 말초 신경 손상, HIV와 연관된 통증, 삼차신경통, 대상포진 후 신경통, 침해수용성 통증, 가족성 홍색사지통증, 원발성 홍색사지통증, 가족성 직장 통증, 섬유근육통, 다발성 경화증(MS)과 연관된 통증으로 이루어진 군으로부터 선택되는 어느 하나인 것을 특징으로 하는, 조성물.These diseases include visceral pain, acute pain, inflammatory pain, postoperative pain, chronic pain, neuropathic pain, cancer pain, chemotherapy pain, trauma pain, surgical pain, childbirth pain, labor pain, persistent pain, peripheral mediated pain, central Mediated pain, chronic headache, migraine, sinus headache, tension headache, phantom limb pain, toothache, peripheral nerve damage, HIV-related pain, trigeminal neuralgia, postherpetic neuralgia, incidence pain, familial erythema The composition characterized in that it is any one selected from the group consisting of pain, primary erythematosis pain, familial rectal pain, fibromyalgia, and pain associated with multiple sclerosis (MS).
PCT/KR2019/014663 2018-11-30 2019-11-01 Composition for preventing or treating pain, containing aconine as active ingredient WO2020111541A1 (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS56120620A (en) * 1980-02-26 1981-09-22 Murayama Keikichi Novel anti-inflammatory agent
JPS63211268A (en) * 1987-02-27 1988-09-02 Sanwa Shiyouyaku Kk Novel aconitine compound, analgesic and anti-inflammatory drug containing said compound as active ingredient
EP0564648A1 (en) * 1991-09-27 1993-10-13 Sanwa Shoyaku Kabushiki Kaisha Novel aconitine compound and analgesic/antiinflammatory agent
JP2000169452A (en) * 1989-12-21 2000-06-20 Sanwa Shoyaku Kk New aconitine-based compound and analgesic antiphlogistic medicine
CN1836661A (en) * 2005-03-25 2006-09-27 成都高新区智爽天然产物科技有限责任公司 Use of aconine in medicine manufacture

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS56120620A (en) * 1980-02-26 1981-09-22 Murayama Keikichi Novel anti-inflammatory agent
JPS63211268A (en) * 1987-02-27 1988-09-02 Sanwa Shiyouyaku Kk Novel aconitine compound, analgesic and anti-inflammatory drug containing said compound as active ingredient
JP2000169452A (en) * 1989-12-21 2000-06-20 Sanwa Shoyaku Kk New aconitine-based compound and analgesic antiphlogistic medicine
EP0564648A1 (en) * 1991-09-27 1993-10-13 Sanwa Shoyaku Kabushiki Kaisha Novel aconitine compound and analgesic/antiinflammatory agent
CN1836661A (en) * 2005-03-25 2006-09-27 成都高新区智爽天然产物科技有限责任公司 Use of aconine in medicine manufacture

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