WO2022080529A1 - Pharmaceutical composition and health functional food for preventing or treating hyposalivation - Google Patents

Pharmaceutical composition and health functional food for preventing or treating hyposalivation Download PDF

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WO2022080529A1
WO2022080529A1 PCT/KR2020/014087 KR2020014087W WO2022080529A1 WO 2022080529 A1 WO2022080529 A1 WO 2022080529A1 KR 2020014087 W KR2020014087 W KR 2020014087W WO 2022080529 A1 WO2022080529 A1 WO 2022080529A1
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Prior art keywords
acid
hyposalivation
pharmaceutical composition
ala
group
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PCT/KR2020/014087
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French (fr)
Korean (ko)
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정정화
김진현
우승훈
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경상대학교병원
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Publication of WO2022080529A1 publication Critical patent/WO2022080529A1/en

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    • 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
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • A61K31/385—Heterocyclic compounds having sulfur as a ring hetero atom having two or more sulfur atoms in the same ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention relates to a pharmaceutical composition and health functional food for preventing or treating hyposalivation.
  • Saliva is a secretion secreted from the salivary glands into the oral cavity. A normal person produces an average of 1-1.5 L of saliva per day. About 99.5% of saliva is water, and the remaining 0.5% is composed of electrolytes, mucin, glycoproteins, enzymes, and antibacterial substances. Saliva lubricates the oral mucosa, protects oral tissues, and is involved in digestion, taste, and remineralization of teeth. In particular, several proteins present in saliva have a profound effect on the oral microbiota due to their antibacterial, antiviral and antifungal actions.
  • the mechanism of salivation can be divided into protein secretion and fluid and electrolyte secretion.
  • protein secretion the sympathetic neurotransmitter norepinephrine binds to the ⁇ -adrenergic receptor present in the cell membrane at the bottom of the secretory terminal cell, resulting in heterotrimeric G-protein (heterotrimeric).
  • G-protein) and adenylyl cyclase are activated.
  • the formation of cyclic adenosine monophosphate is catalyzed, and when protein kinase A is activated, the secretory granules present in the cell membrane secrete the protein toward the lumen by exocytosis.
  • secretion of liquid and electrolyte mainly occurs by binding of acetylcholine, a parasympathetic neurotransmitter, to muscarinic receptors.
  • an agonist binds to a muscarinic receptor
  • heterotrimeric G-protein and phospholipase C are sequentially activated and ultimately inositol triphosphate , IP3) is promoted.
  • IP3 releases Ca 2+ stored in the cell, and the Cl - channel on the luminal side and K + channel on the bottom side are opened by the increased Ca 2+ concentration to open Na + /K + /2Cl - co-
  • the co-transporter is activated.
  • aquaporin 5 (AQP5) protein present in the vesicle moves to the luminal cell membrane.
  • the increased luminal Cl ⁇ is balanced along with Na + moving into the lumen through the closed junction, resulting in an osmotic gradient.
  • the osmotic gradient moves water from the cell towards the lumen through the closed junction with AQP5.
  • aquaporin as a water channel protein is found in the cornea, kidney, liver, skin, and the like.
  • AQP5 is a major biomarker for salivation.
  • research results have been published that the amount of saliva secretion decreased by more than 60% in AQP5 knockout mice.
  • Xerostomia is the most common clinical symptom associated with salivary glands. Dry mouth is diagnosed when the amount of saliva secreted during non-stimulation is less than 0.1 ml per minute. The causes of dry mouth are mainly side effects caused by drugs such as antidepressants, antidiabetics, and antiallergy. Dry mouth caused by natural aging is caused by a decrease in the parenchymal tissue and replacement by adipose tissue or a decrease in the volume of the alveoli. When the amount of saliva in the oral cavity is reduced, tooth decay, acid erosion, oral candidiasis, dysgeusia, dysphagia, oral dysesthesia, and intraoral halitosis may occur. accompanying
  • Representative treatments for dry mouth currently used include parasympathomimetic pilocarpine, cevimeline, and bethanechol.
  • these treatments have the disadvantage of causing side effects such as sweating, vomiting, vasodilation, diarrhea, and bronchospasm. Therefore, it is a very important task to prevent and treat dry mouth based on natural products with few side effects and high safety features.
  • An object of the present invention is to provide a pharmaceutical composition for preventing or treating hyposalivation, comprising ⁇ -Lipoic acid or a pharmaceutically acceptable salt thereof.
  • Another object of the present invention is to provide a health functional food for preventing or improving hyposalivation, comprising alpha-Lipoic acid or a pharmaceutically acceptable salt thereof.
  • a pharmaceutical composition for preventing or treating hyposalivation comprising ⁇ -Lipoic acid or a pharmaceutically acceptable salt thereof.
  • Health functional food for preventing or improving hyposalivation, including ⁇ -Lipoic acid or a pharmaceutically acceptable salt thereof.
  • the pharmaceutical composition comprising alpha lipoic acid of the present invention can achieve the prevention or treatment effect of hyposalivation by improving the damage to the salivary gland, and the health functional food of the present invention can achieve the prevention or improvement effect of hyposalivation.
  • FIG. 1 is a diagram showing improvement of weight loss and damaged saliva secretion due to radioiodine (RI) treatment.
  • FIG. 2 is a diagram confirming the effect of ALA on thyroid function after RI treatment.
  • 3 and 4 are diagrams confirming the pathological results after RI treatment.
  • 5 and 6 are diagrams confirming the effect of ALA on increased cell death and aging after RI treatment.
  • FIG. 7 is a diagram showing that the administration of ALA improves the radioiodine (RI) treatment-induced decrease in AQP5 expression.
  • FIG 8 and 9 are diagrams showing planar whole body images and dynamics of 99m Tc pertechnetate absorption and excretion.
  • compositions for preventing or treating hyposalivation comprising alpha-lipoic acid or a pharmaceutically acceptable salt thereof.
  • the alpha lipoic acid is a compound represented by the following Chemical Formula 1, the chemical formula is C 8 H 14 O 2 S 2 , the molar mass is 206.33 g/mol, and the IUPAC name is 5-(1,2-dithiolan-3-yl)pentanoic acid am:
  • the alpha lipoic acid may be derived from nature or may be synthesized using a known organic synthesis method.
  • the alpha lipoic acid may be a non-protein compound, a peptide, an extract of a plant-derived tissue or cell, or a product obtained by culturing a microorganism (eg, bacteria or fungi, and particularly yeast).
  • a microorganism eg, bacteria or fungi, and particularly yeast
  • the pharmaceutical composition according to the present invention may be provided as a pharmaceutical composition including an active ingredient alone or one or more pharmaceutically acceptable carriers, excipients or diluents.
  • “Pharmaceutically acceptable” in the present invention means exhibiting a property that is not toxic to cells or humans exposed to the composition.
  • the term “pharmaceutically acceptable salt” refers to a salt prepared using a specific compound according to the present invention and a relatively non-toxic acid or base.
  • base addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of the base in neat solution or in a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, ammonium, organic amines, or magnesium or similar salts.
  • acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent.
  • Pharmaceutically acceptable acid addition salts include salts of inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogen carbonate ion, phosphoric acid, phosphate monohydrogen ion, phosphate dihydrogen ion, sulfuric acid, hydrogen sulfate ion, hydroiodic acid or phosphorous acid; and salts of organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid and salts of amino acids (eg, arginine) and salts of organic acids such as glucuronic acid are also included.
  • inorganic acids such as hydrochloric acid, hydrobromic acid,
  • the pharmaceutically acceptable salt of the present invention can be synthesized from a parent compound containing an acidic or basic moiety by a conventional chemical method.
  • such salts are prepared by reacting the free acid or form of the base of these compounds with a stoichiometrically appropriate amount of the base or acid, in water or in an organic solvent or in a mixture of the two.
  • non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.
  • the pharmaceutical composition of the present invention may be provided by mixing with a conventionally known composition for preventing or treating hyposalivation. That is, the pharmaceutical composition of the present invention may be administered in parallel with a known compound having an effect of preventing or treating hyposalivation.
  • administration means introducing a predetermined substance to an individual by an appropriate method
  • individual means all living things, such as mice, mice, and livestock, including humans, that may have hyposalivation. As a specific example, it may be a mammal including a human.
  • the pharmaceutical composition of the present invention may additionally include a composition having a known preventive or therapeutic effect on hyposalivation.
  • compositions for preventing or treating hyposalivation include Java turmeric ( Curcumaxanthorrhiza ) extract, polyglutamic acid and Aspalathus linearis ( Aspalathus linearis ) extract, Hepatica oleracea extract, warthog extract, and the like, but are limited thereto. not.
  • the route of administration of the pharmaceutical composition is, but not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal.
  • composition of the present invention may be administered orally or parenterally, and when administered parenterally, it is preferable to select an injection method for external application or intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection. , but is not limited thereto.
  • the preferred dosage of the composition of the present invention varies depending on the condition and weight of the patient, the degree of disease, the drug form, the route and duration of administration, but may be appropriately selected by those skilled in the art. However, for a desirable effect, the composition is preferably administered at 0.01 to 1000 mg/kg/day, preferably at 0.1 to 500 mg/kg/day, but is not limited thereto. The administration may be administered once a day, or divided into several administrations. The above dosage does not limit the scope of the present invention in any way.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose. Alternatively, it is prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.
  • Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, etc.
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories.
  • Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.
  • a suppository base witepsol, macrogol, tween 61, cacao butter, laurin, glycero geratin and the like can be used.
  • the hyposalivation may be caused by various causes, for example, may be caused by damage to the salivary gland, but is not limited thereto.
  • the salivary gland damage may also be caused by various causes, for example, it may be caused by radioiodine treatment, but is not limited thereto.
  • the alpha lipoic acid can increase the amount of salivary secretion by restoring the damage to the salivary glands, specifically, can increase the weight of the salivary glands reduced due to damage to the salivary glands again, and specifically decrease the salivary gland-related factor of aquaporin 5. It may increase expression, but is not limited thereto.
  • the alpha lipoic acid can restore damaged parasympathetic nerves, and specifically can increase the expression of glial cell-derived neurotrophic factor family receptor 2 (GFR2), which is a marker of parasympathetic nerves. and may increase the expression of brain-derived neurotrophic factor (BDNF) and neurturin related to parasympathetic nerves, but is not limited thereto.
  • GFR2 glial cell-derived neurotrophic factor family receptor 2
  • BDNF brain-derived neurotrophic factor
  • neurturin related to parasympathetic nerves but is not limited thereto.
  • the alpha lipoic acid can regenerate the salivary glands, specifically hedgehog (Hedgehog, Hh) signaling related to the regeneration of the salivary gland Sonic hedgehog (Sonic Hedgehog, Shh) and its receptor Patched (Patched, Ptch) It is possible to increase the expression of the gene and may increase the expression of Sca-1, a stem cell-related marker related to regeneration of the salivary gland, but is not limited thereto.
  • the present invention relates to a health functional food for preventing or improving hyposalivation, comprising ⁇ -Lipoic acid or a pharmaceutically acceptable salt thereof.
  • the hyposalivation may be caused by damage to the salivary glands, and the damage to the salivary glands may be caused by radioiodine treatment, but is not limited thereto.
  • the efficacy, mechanism of action, etc. of the alpha lipoic acid may be within the above-mentioned range.
  • food pharmaceutically acceptable means exhibiting a property that is not toxic to cells or humans exposed to the compound.
  • “food pharmaceutically acceptable salt” means a salt prepared using a specific compound according to the present invention and a relatively non-toxic acid or base, and may be within the above-described range for “salt”.
  • the health functional food of the present invention may be formulated into one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid formulations, further comprising one or more of carriers, diluents, excipients and additives.
  • Foods to which the extract of the present invention can be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gums, tea, vitamin complexes, health functional foods, and the like.
  • Additives that may be further included in the present invention include natural carbohydrates, flavoring agents, nutrients, vitamins, minerals (electrolytes), flavoring agents (synthetic flavoring agents, natural flavoring agents, etc.), coloring agents, fillers (cheese, chocolate, etc.), At least one component selected from the group consisting of facic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, antioxidants, glycerin, alcohols, carbonation agents, and pulp may be used. .
  • natural carbohydrates examples include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents tacartin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
  • the health functional food of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, colorants and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
  • the composition according to the present invention may contain the pulp for the production of natural fruit juices and vegetable beverages. These components may be used independently or in combination.
  • carrier examples include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium Silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and at least one selected from the group consisting of mineral oil is preferably used.
  • the health functional food of the present invention it is prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, and surfactant usually used.
  • a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, and surfactant usually used.
  • Salivary function was assessed by measuring salivary secretion. After injection of pilocarpine (0.01 mL/g body weight i.p.; Isopto Carpine; Alcon Korea Ltd., Seoul, Korea), saliva discharge was collected from the mouth for 5 minutes after 8 minutes. After measuring the total body weight of the mice, the total amount and volume of saliva accumulated for 10 minutes after pilocarpine injection into a fresh tube was normalized to body weight. The needle delay time and flow rate were measured. Salivary flow rate (total needle weight divided by collection time) and delay time (time from stimulation to onset of salivation) were calculated.
  • Tissues were fixed in 4% paraformaldehyde in 0.1M PBS, embedded in paraffin, and cut into 5 ⁇ m. Sections were stained with HE. Histopathological damage in HE staining was scored by grading the number of acinar cells with cytoplasmic vacuoles in a ⁇ 200 magnification field. 0, 0-1; 1, 2-5; 2, 5-10; 3, 10-15; 4, 15-20; 5, > 20. Sections were stained with MT (Masson's Trichrome kit, Sigma Diagnostics, St. Louis, MO, USA) to analyze the degree of collagen deposition. Randomly selected SG (salivary gland) fields at 400x magnification were evaluated in each mouse, and the density of trichrome-positive signals was analyzed using NIS-Elements BR 3.2 (Nikon, Japan).
  • SG was immediately put in 4% paraformaldehyde at room temperature, embedded in paraffin, and then cut to a thickness of 5 ⁇ m. SGs were stained with Masson's trichrome (MT) and hematoxylin and eosin (H&E). Apoptosis of submaxillary gland tissues was determined by TUNEL analysis using the ApopTag Plus In Situ Apoptosis Kit (Chemicon Int., Temecula, CA, USA). TUNEL-positive cells were detected at ⁇ 400 magnification, and TUNEL-positive cells were counted in 10 random high-power fields. TUNEL analysis was performed at 30 and 90 days post RI exposure.
  • MT Masson's trichrome
  • H&E hematoxylin and eosin
  • Sections were visualized by light microscopy and digital images were captured and analyzed. Data were analyzed as signal intensity using NIS-Elements BR 3.2 (Nikon, Japan) in 10 random fields and accounted for by fold change. Fold change was calculated as the ratio of the final value of each group to the value of the control group at day 30 (set as "1").
  • Whole-body SPECT images were obtained using a large field-of-view rotating gamma camera equipped with four multi-pinhole collimators. Acquisition parameters are: 24 projections over 360°, circular trajectory, and a total acquisition time of 6 minutes (4 seconds per projection). The tomographic images were reconstructed using an iterative reconstruction algorithm.
  • ALA improves weight loss and impaired salivation due to radioiodine exposure.
  • the average body weight of the RI-treated mouse group was significantly lower than that of the control group at 90 days, and the body weights of the RI+ALA and ALA groups were similar to those of the control group ( FIG. 1A ).
  • the salivary flow rate of the RI+ALA-treated group was higher than that of the RI-only group at 30 and 90 days after RI (Fig. 1B).
  • salivation delay was improved in the RI+ALA group compared to the RI alone group (FIG. 1C).
  • TSH thyroid stimulating hormone
  • HE hematoxylin and eosin
  • MT Masson trichrome
  • Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assay was performed to determine the effect of ALA on RI-induced apoptosis.
  • a faint signal was found in both the control and ALA-only groups at 30 and 90 days after RI treatment; They were more abundant in acinar than ductal cells and more pronounced at 30 days than at 90 days ( Figures 5A, 5B).
  • the positive signal was significantly increased in the RI-only group compared to the control and ALA-only groups.
  • the total number of apoptotic cells was significantly lower in the ALA+RI group compared to the RI alone group at both 30 and 90 days after RI treatment (Fig. 5C).
  • ALA prevents RI-induced cellular senescence in SGs.
  • SA- ⁇ -gal senescence-associated ⁇ -galactosidase
  • ALA restores the radioiodine-induced decrease in AQP5 expression.
  • ALA ameliorates RI-induced acupuncture dysfunction.
  • Acupuncture function was examined after RI treatment using single-photon emission computed tomography (SPECT).
  • SPECT single-photon emission computed tomography
  • the degree of 99m Tc pertechnetate clearance was much lower in the RI alone group at 110 and 120 minutes.
  • the excretion of 99m Tc pertechnetate in the ALA-only group was similar to that of the control group ( FIGS. 8A and 8B ).
  • the level of 99m Tc pertechnetate clearance was significantly lower in the RI-only group compared to the other groups at 70, 90, 100, 110 and 120 min.
  • the excretion of 99m Tc pertechnetate in the ALA-only group was better than in the control group ( FIGS. 9A and 9B ).

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Abstract

The present invention relates to a pharmaceutical composition and a health functional food comprising alpha lipoic acid or a salt thereof, which alleviate damage to salivary glands to achieve an effect of preventing or treating hyposalivation, and more specifically, exhibit an excellent effect in alleviating damage to salivary glands caused by radioiodine (RI) treatment.

Description

침분비 저하증 예방 또는 치료용 약학적 조성물 및 건강기능식품Pharmaceutical composition and health functional food for preventing or treating hyposalivation
본 발명은 침분비 저하증 예방 또는 치료용 약학적 조성물 및 건강기능식품에 관한 것이다.The present invention relates to a pharmaceutical composition and health functional food for preventing or treating hyposalivation.
침은 침샘에서 구강으로 분비되는 분비액으로 정상인의 경우 일일 평균 1 ~ 1.5 L의 침이 생성된다. 침의 약 99.5 %는 수분이며 나머지 0.5 %는 전해질, 점액소, 당단백질, 효소, 항균성 물질 등으로 구성된다. 침은 구강 점막을 윤활하게 하고 구강조직을 보호하며, 소화, 미각, 치아의 재무기물화(remineralization)등에 관여한다. 특히, 침에 존재하는 여러 단백질들은 항박테리아, 항바이러스와 항곰팡이 작용으로 구강미생물총(oral microbiota)에 지대한 영향을 미친다.Saliva is a secretion secreted from the salivary glands into the oral cavity. A normal person produces an average of 1-1.5 L of saliva per day. About 99.5% of saliva is water, and the remaining 0.5% is composed of electrolytes, mucin, glycoproteins, enzymes, and antibacterial substances. Saliva lubricates the oral mucosa, protects oral tissues, and is involved in digestion, taste, and remineralization of teeth. In particular, several proteins present in saliva have a profound effect on the oral microbiota due to their antibacterial, antiviral and antifungal actions.
침 분비기전은 단백질 분비와 액체와 전해질의 분비로 나눌 수 있다. 단백질 분비의 경우, 분비종말부세포의 바닥 쪽 세포막에 존재하는 β-아드레날린작동성 수용체(β-adrenergic receptor)에 교감신경전달물질인 노르에피네프린(norepinephrine)이 결합함으로써 헤테로트리머릭 G-프로테인(heterotrimeric G-protein)과 아데닐일 고리화효소(adenylyl cyclase)가 활성화된다. 이로 인해 고고리형 아데노신1인산(cyclic adenosine monophosphate)의 형성이 촉매되어 단백질 키나아제 A(protein kinase A)가 활성화되면, 세포막에 존재하는 분비과립들이 세포외유출 방법으로 단백질을 내강 쪽으로 분비하게 된다. 한편, 액체와 전해질의 분비는 muscarinic receptor에 부교감신경전달물질인 아세틸콜린(acetylcholine)의 결합에 의해 주로 일어난다. 무스카린 수용체(Muscarinic receptor)에 아고니스트(agonist)가 결합하게 되면 헤테로트리머릭 G-프로테인(heterotrimeric G-protein)과 포스포리파아제 C(phospholipase C)가 차례로 활성화되어 궁극적으로 이노시톨 트리포스페이트(inositol triphosphate, IP₃)의 형성이 촉진된다. IP₃는 세포 내에 저장되어 있는 Ca 2+을 방출하고, 증가된 Ca 2+의 농도에 의해 내강 쪽에 존재하는 Cl - 통로와 바닥 쪽에 존재하는 K + 통로가 열려 Na +/K +/2Cl - 공동-운반체(co-transporter)가 활성화된다. 또한, 소낭에 존재하는 아쿠아포린 5(aquaporin 5, AQP5) 단백질이 내강 쪽 세포막으로 이동한다. 증가된 내강 쪽의 Cl -은 폐쇄연접을 통해 내강으로 이동하는 Na +을 따라 균형을 맞추고 그 결과 삼투압 기울기(osmotic gradient)가 발생한다. 삼투압 기울기는 AQP5와 폐쇄연접을 통해 물을 세포에서 내강 쪽으로 이동시킨다. The mechanism of salivation can be divided into protein secretion and fluid and electrolyte secretion. In the case of protein secretion, the sympathetic neurotransmitter norepinephrine binds to the β-adrenergic receptor present in the cell membrane at the bottom of the secretory terminal cell, resulting in heterotrimeric G-protein (heterotrimeric). G-protein) and adenylyl cyclase are activated. As a result, the formation of cyclic adenosine monophosphate is catalyzed, and when protein kinase A is activated, the secretory granules present in the cell membrane secrete the protein toward the lumen by exocytosis. On the other hand, secretion of liquid and electrolyte mainly occurs by binding of acetylcholine, a parasympathetic neurotransmitter, to muscarinic receptors. When an agonist binds to a muscarinic receptor, heterotrimeric G-protein and phospholipase C are sequentially activated and ultimately inositol triphosphate , IP₃) is promoted. IP₃ releases Ca 2+ stored in the cell, and the Cl - channel on the luminal side and K + channel on the bottom side are opened by the increased Ca 2+ concentration to open Na + /K + /2Cl - co- The co-transporter is activated. In addition, aquaporin 5 (AQP5) protein present in the vesicle moves to the luminal cell membrane. The increased luminal Cl − is balanced along with Na + moving into the lumen through the closed junction, resulting in an osmotic gradient. The osmotic gradient moves water from the cell towards the lumen through the closed junction with AQP5.
한편, 아쿠아포린(aquaporin)은 물 통로 단백질로서 각막, 신장, 간, 피부 등에서 발견된다. 아쿠아포린(aquaporin)은 AQP0~12까지 아과(subfamily)가 다양하게 존재하는데, AQP5는 침 분비에 주요 바이오마커(biomarker)이다. 특히, AQP5 넉아웃 마이스(AQP5 knockout mice)에서 침 분비량이 60 % 이상 감소했다는 연구결과가 발표된 바 있다.On the other hand, aquaporin (aquaporin) as a water channel protein is found in the cornea, kidney, liver, skin, and the like. There are various subfamily (subfamily) of aquaporin from AQP0 to 12, and AQP5 is a major biomarker for salivation. In particular, research results have been published that the amount of saliva secretion decreased by more than 60% in AQP5 knockout mice.
구강건조증(Xerostomia)은 침샘과 관련하여 가장 흔한 임상적 증상이다. 비자극시에 분비되는 침의 양이 분당 0.1 ㎖ 이하인 경우 구강건조증으로 진단된다. 구강건조증의 발병원인은 주로 항우울증, 항당뇨, 항알레르기 등의 약물에 의한 부작용이다. 자연 노화에 의한 구강건조증은 의 실질조직이 감소하고 지방조직으로 대체되거나 샘꽈리 용적이 감소하여 나타난다. 구강 내 침 양이 감소되면 충치, 산식증(acid erosion), 구강칸디다증(oral candidiasis), 미각장애(dysgeusia), 연하곤란(dysphagia), 구강 감각장애(oral dysesthesia), 구취(intraoral halitosis) 등이 동반된다.Xerostomia is the most common clinical symptom associated with salivary glands. Dry mouth is diagnosed when the amount of saliva secreted during non-stimulation is less than 0.1 ml per minute. The causes of dry mouth are mainly side effects caused by drugs such as antidepressants, antidiabetics, and antiallergy. Dry mouth caused by natural aging is caused by a decrease in the parenchymal tissue and replacement by adipose tissue or a decrease in the volume of the alveoli. When the amount of saliva in the oral cavity is reduced, tooth decay, acid erosion, oral candidiasis, dysgeusia, dysphagia, oral dysesthesia, and intraoral halitosis may occur. accompanying
현재 사용되고 있는 대표적인 구강건조증 치료제는 부교감신경작용제(parasympathomimetic)인 필로카르핀(pilocarpine), 세비멜린(cevimeline), 베탄콜(bethanechol) 등이 있다. 하지만, 이러한 치료제들은 발한, 구토, 혈관확장, 설사, 기관지경련(bronchospasm)과 같은 부작용을 유발시키는 단점이 있다. 따라서 적은 부작용과 높은 안전성의 특징을 지닌 천연물을 기반으로 하여 구강건조증을 예방하고 치료하는 것은 매우 중요한 과제이다.Representative treatments for dry mouth currently used include parasympathomimetic pilocarpine, cevimeline, and bethanechol. However, these treatments have the disadvantage of causing side effects such as sweating, vomiting, vasodilation, diarrhea, and bronchospasm. Therefore, it is a very important task to prevent and treat dry mouth based on natural products with few side effects and high safety features.
한편, 방사성요오드 치료 이후에는 침샘이 손상되며, 이에 따라 침 분비량이 적어진다. 이에, 침샘의 손상을 개선함에 따라 침분비량을 회복할 수 있는 조성물이 요구된다.On the other hand, after radioiodine treatment, the salivary glands are damaged, and the amount of saliva secretion decreases accordingly. Accordingly, there is a need for a composition capable of recovering the amount of salivary secretion by improving the damage to the salivary glands.
본 발명은 알파리포산(α-Lipoic acid) 또는 이의 약학적으로 허용되는 염을 포함하는 침분비 저하증 예방 또는 치료용 약학적 조성물을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a pharmaceutical composition for preventing or treating hyposalivation, comprising α-Lipoic acid or a pharmaceutically acceptable salt thereof.
또한 본 발명은 알파리포산(α-Lipoic acid) 또는 이의 식품학적으로 허용되는 염을 포함하는 침분비 저하증 예방 또는 개선용 건강기능식품을 제공하는 것을 목적으로 한다.Another object of the present invention is to provide a health functional food for preventing or improving hyposalivation, comprising alpha-Lipoic acid or a pharmaceutically acceptable salt thereof.
1. 알파리포산(α-Lipoic acid) 또는 이의 약학적으로 허용되는 염을 포함하는 침분비 저하증 예방 또는 치료용 약학적 조성물.1. A pharmaceutical composition for preventing or treating hyposalivation, comprising α-Lipoic acid or a pharmaceutically acceptable salt thereof.
2. 위 1에 있어서, 상기 침분비 저하증은 침샘 손상으로부터 유발되는 것인, 약학적 조성물.2. The pharmaceutical composition of 1 above, wherein the hyposalivation is caused by damage to the salivary glands.
3. 위 2에 있어서, 상기 침샘 손상은 방사성요오드 치료로 유발되는 것인, 약학적 조성물.3. The pharmaceutical composition of 2 above, wherein the salivary gland damage is induced by radioiodine treatment.
4. 알파리포산(α-Lipoic acid) 또는 이의 식품학적으로 허용되는 염을 포함하는 침분비 저하증 예방 또는 개선용 건강기능식품.4. Health functional food for preventing or improving hyposalivation, including α-Lipoic acid or a pharmaceutically acceptable salt thereof.
5. 위 4에 있어서, 상기 침분비 저하증은 침샘 손상으로부터 유발되는 것인, 건강기능식품.5. The above 4, wherein the hyposalivation is caused by damage to the salivary gland, health functional food.
6. 위 5에 있어서, 상기 침샘 손상은 방사성요오드 치료로 유발되는 것인, 건강기능식품.6. The health functional food according to the above 5, wherein the salivary gland damage is induced by radioiodine treatment.
본 발명의 알파리포산을 포함하는 약학적 조성물은 침샘의 손상을 개선함으로써 침분비 저하증의 예방 또는 치료 효과를 달성할 수 있고, 본 발명의 건강기능식품은 침분비 저하증의 예방 또는 개선 효과를 달성할 수 있다.The pharmaceutical composition comprising alpha lipoic acid of the present invention can achieve the prevention or treatment effect of hyposalivation by improving the damage to the salivary gland, and the health functional food of the present invention can achieve the prevention or improvement effect of hyposalivation. can
도 1 은 방사성요오드(RI) 치료로 인한 체중 손실 및 손상된 침 분비를 개선하는 것을 나타낸 도이다.1 is a diagram showing improvement of weight loss and damaged saliva secretion due to radioiodine (RI) treatment.
도 2는 RI 치료 후 갑상선 기능에 대한 ALA의 효과를 확인한 도이다.2 is a diagram confirming the effect of ALA on thyroid function after RI treatment.
도 3 및 4는 RI 치료 후 병리학적 결과를 확인한 도이다.3 and 4 are diagrams confirming the pathological results after RI treatment.
도 5 및 6은 ALA가 RI 치료 후 증가된 세포 사멸과 노화에 미치는 영향을 확인한 도이다.5 and 6 are diagrams confirming the effect of ALA on increased cell death and aging after RI treatment.
도 7은 ALA의 투여가 방사성요오드(RI) 치료 유발 AQP5 발현 감소를 개선하는 것을 나타내는 도이다.7 is a diagram showing that the administration of ALA improves the radioiodine (RI) treatment-induced decrease in AQP5 expression.
도 8 및 9는 99mTc pertechnetate 흡수 및 배출의 평면 전신 이미지 및 역학(dynamics)을 나타낸 도이다.8 and 9 are diagrams showing planar whole body images and dynamics of 99m Tc pertechnetate absorption and excretion.
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
알파리포산(α-Lipoic acid) 또는 이의 약학적으로 허용되는 염을 포함하는 침분비 저하증 예방 또는 치료용 약학적 조성물에 관한 것이다.It relates to a pharmaceutical composition for preventing or treating hyposalivation, comprising alpha-lipoic acid or a pharmaceutically acceptable salt thereof.
상기 알파리포산은 하기 화학식 1로 표시되는 화합물로서, 화학식은 C 8H 14O 2S 2, 몰질량은 206.33g/mol, IUPAC 명명은 5-(1,2-dithiolan-3-yl)pentanoic acid이다:The alpha lipoic acid is a compound represented by the following Chemical Formula 1, the chemical formula is C 8 H 14 O 2 S 2 , the molar mass is 206.33 g/mol, and the IUPAC name is 5-(1,2-dithiolan-3-yl)pentanoic acid am:
[화학식 1][Formula 1]
Figure PCTKR2020014087-appb-img-000001
Figure PCTKR2020014087-appb-img-000001
상기 알파리포산은 천연으로부터 유래될 수도 있고, 공지의 유기 합성 방법을 이용하여 합성될 수도 있다.The alpha lipoic acid may be derived from nature or may be synthesized using a known organic synthesis method.
상기 알파리포산은 비단백질 화합물, 펩티드, 식물 유래 조직이나 세포의 추출물, 미생물(예를 들어 세균류 또는 진균류, 그리고 특히 효모)의 배양으로 얻어진 생산물일 수 있다.The alpha lipoic acid may be a non-protein compound, a peptide, an extract of a plant-derived tissue or cell, or a product obtained by culturing a microorganism (eg, bacteria or fungi, and particularly yeast).
본 발명에 따른 약학적 조성물은 유효성분을 단독으로 포함하거나, 하나 이상의 약학적으로 허용되는 담체, 부형제 또는 희석제를 포함하여 약학적 조성물로 제공될 수 있다.The pharmaceutical composition according to the present invention may be provided as a pharmaceutical composition including an active ingredient alone or one or more pharmaceutically acceptable carriers, excipients or diluents.
본 발명에서 "약학적으로 허용되는"이란 상기 조성물에 노출되는 세포나 인간에게 독성이 없는 특성을 나타내는 것을 의미한다."Pharmaceutically acceptable" in the present invention means exhibiting a property that is not toxic to cells or humans exposed to the composition.
본 발명의 용어 "약학적으로 허용되는 염"이란, 본 발명에 따른 특정 화합물과 비교적 무독성인 산 또는 염기를 이용해서 조제되는 염을 의미한다. 본 발명의 화합물이 상대적으로 산성 관능기를 포함할 때, 순수 용액 또는 적합한 불활성 용매 중에서 충분한 양의 염기를 이러한 화합물의 중성 형태와 접촉시킴으로써 염기 부가염을 얻을 수 있다. 약학적으로 허용되는 염기 부가염은 나트륨, 칼륨, 칼슘, 암모늄, 유기 아민, 혹은 마그네슘의 염 또는 유사한 염이 포함된다. 본 발명의 화합물이 상대적으로 염기성 관능기를 포함할 때, 순수 용액 또는 적합한 불활성 용매 중에서 충분한 양의 산을 이러한 화합물의 중성 형태와 접촉시킴으로써 산 부가염을 얻을 수 있다. 약학적으로 허용되는 산 부가염은 염산, 브롬화 수소산, 질산, 탄산, 탄산 수소 이온, 인산, 인산 1수소 이온, 인산 2수소 이온, 황산, 황산 수소 이온, 요오드화 수소산 또는 아인산 등의 무기산의 염, 그리고 아세트산, 프로피온산, 이소부티르산, 말레산, 말론산, 안식향산, 숙신산, 수베르산, 푸마르산, 락트산, 만델산, 프탈산, 벤젠술폰산, p-톨릴술폰산, 구연산, 주석산, 메탄술폰산 등의 유기산의 염을 들 수 있고, 나아가 아미노산(예를 들면 아르기닌 등)의 염 및 글루쿠론산 등의 유기산의 염도 포함된다.As used herein, the term “pharmaceutically acceptable salt” refers to a salt prepared using a specific compound according to the present invention and a relatively non-toxic acid or base. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of the base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, ammonium, organic amines, or magnesium or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include salts of inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogen carbonate ion, phosphoric acid, phosphate monohydrogen ion, phosphate dihydrogen ion, sulfuric acid, hydrogen sulfate ion, hydroiodic acid or phosphorous acid; and salts of organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid and salts of amino acids (eg, arginine) and salts of organic acids such as glucuronic acid are also included.
본 발명의 약학적으로 허용되는 염은 산성 또는 염기성 부분을 포함하는 모체 화합물로부터 통상적인 화학적 방법으로 합성할 수 있다. 일반적으로 이러한 염은 수중 또는 유기 용매 중 또는 이 2종의 혼합물 중에서, 이들 화합물의 유리산 또는 염기의 형태를 화학량론적으로 적량인 염기 또는 산과 반응시켜서 조제된다. 일반적으로 에테르, 아세트산에틸, 에탄올, 이소프로판올 또는 아세토니트릴 등의 비수성 매질이 바람직하다.The pharmaceutically acceptable salt of the present invention can be synthesized from a parent compound containing an acidic or basic moiety by a conventional chemical method. In general, such salts are prepared by reacting the free acid or form of the base of these compounds with a stoichiometrically appropriate amount of the base or acid, in water or in an organic solvent or in a mixture of the two. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.
나아가 본 발명의 약학적 조성물은 종래에 알려져 있는 침분비 저하증 예방 또는 치료용 조성물과 혼합하여 제공될 수도 있다. 즉, 본 발명의 약학적 조성물은 침분비 저하증 예방 또는 치료 효과를 가지는 공지의 화합물과 병행하여 투여할 수 있다Furthermore, the pharmaceutical composition of the present invention may be provided by mixing with a conventionally known composition for preventing or treating hyposalivation. That is, the pharmaceutical composition of the present invention may be administered in parallel with a known compound having an effect of preventing or treating hyposalivation.
본 발명에서 "투여"란 적절한 방법으로 개체에게 소정의 물질을 도입하는 것을 의미하며, "개체"란 침분비 저하증을 보유할 수 있는 인간을 포함한 쥐, 생쥐, 가축 등의 모든 생물을 의미한다. 구체적인 예로, 인간을 포함한 포유동물일 수 있다.In the present invention, "administration" means introducing a predetermined substance to an individual by an appropriate method, and "individual" means all living things, such as mice, mice, and livestock, including humans, that may have hyposalivation. As a specific example, it may be a mammal including a human.
필요에 따라, 본 발명의 약학적 조성물은 공지의 침분비 저하증 예방 또는 치료 효과를 가진 조성물을 추가적으로 포함할 수 있다.If necessary, the pharmaceutical composition of the present invention may additionally include a composition having a known preventive or therapeutic effect on hyposalivation.
이러한 침분비 저하증 예방 또는 치료용 조성물로는 자바강황( Curcumaxanthorrhiza) 추출물, 폴리글루타민산 및 아스팔라투스-리네아리스(Aspalathus linearis) 추출물, 노루궁뎅이버섯 추출물, 씀바귀 추출물 등을 들 수 있으나, 이에 제한되는 것은 아니다.Examples of the composition for preventing or treating hyposalivation include Java turmeric ( Curcumaxanthorrhiza ) extract, polyglutamic acid and Aspalathus linearis ( Aspalathus linearis ) extract, Hepatica oleracea extract, warthog extract, and the like, but are limited thereto. not.
본 발명에 있어서, 약학적 조성물의 투여 경로는 이들로 한정되는 것은 아니지만 구강, 정맥내, 근육내, 동맥내, 골수내, 경막내, 심장내, 경피, 피하, 복강내, 비강내, 장관, 국소, 설하 또는 직장이 포함된다.In the present invention, the route of administration of the pharmaceutical composition is, but not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal.
본 발명의 조성물은 경구 또는 비경구 투여할 수 있으며, 비경구 투여시 피부 외용 또는 복강내주사, 직장내주사, 피하주사, 정맥주사, 근육내 주사 또는 흉부내 주사 주입방식을 선택하는 것이 바람직하며, 이에 한정되는 것은 아니다.The composition of the present invention may be administered orally or parenterally, and when administered parenterally, it is preferable to select an injection method for external application or intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection. , but is not limited thereto.
본 발명의 조성물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 바람직한 효과를 위해서, 상기 조성물은 0.01~1000mg/kg/day로, 바람직하게는 0.1~500㎎/kg/day로 투여하는 것이 바람직하나 이에 한정되지 않는다. 상기 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The preferred dosage of the composition of the present invention varies depending on the condition and weight of the patient, the degree of disease, the drug form, the route and duration of administration, but may be appropriately selected by those skilled in the art. However, for a desirable effect, the composition is preferably administered at 0.01 to 1000 mg/kg/day, preferably at 0.1 to 500 mg/kg/day, but is not limited thereto. The administration may be administered once a day, or divided into several administrations. The above dosage does not limit the scope of the present invention in any way.
제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calciumcarbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜 (propyleneglycol), 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔 (witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로 제라틴 등이 사용될 수 있다.In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose. Alternatively, it is prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients, for example, wetting agents, sweeteners, fragrances, preservatives, etc. may be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin, glycero geratin and the like can be used.
상기 침분비 저하증은 다양한 원인으로 유발되는 것일 수 있고, 예를 들어, 침샘 손상으로부터 유발되는 것일 수 있으나, 이에 제한되는 것은 아니다. 또한, 상기 침샘 손상 역시 다양한 원인으로 유발되는 것일 수 있고, 예를 들어, 방사성요오드 치료로 유발되는 것일 수 있으나, 이에 제한되는 것은 아니다.The hyposalivation may be caused by various causes, for example, may be caused by damage to the salivary gland, but is not limited thereto. In addition, the salivary gland damage may also be caused by various causes, for example, it may be caused by radioiodine treatment, but is not limited thereto.
상기 알파리포산은 상기 침샘의 손상을 회복함으로써 침 분비량을 증가시킬 수 있고, 구체적으로 침샘 손상으로 인한 침샘의 감소된 중량을 다시 증가시킬 수 있고, 구체적으로 감소된 침분비 관련 인자인 아쿠아포린5의 발현을 증가시킬 수 있으나, 이에 제한되는 것은 아니다.The alpha lipoic acid can increase the amount of salivary secretion by restoring the damage to the salivary glands, specifically, can increase the weight of the salivary glands reduced due to damage to the salivary glands again, and specifically decrease the salivary gland-related factor of aquaporin 5. It may increase expression, but is not limited thereto.
또한, 상기 알파리포산은 손상된 부교감 신경을 회복시킬 수 있고, 구체적으로 부교감신경의 마커인 신경아교세포 유래 신경영양 인자 패밀리 수용체 2(glial cell-derived neurotrophic factor family receptor 2, GFR2)의 발현을 증가시킬 수 있고, 부교감 신경과 관련된 뇌 유발 신경영양 인자(brain-derived neurotrophic factor, BDNF) 및 뉴투린(neurturin)의 발현을 증가시킬 수 있으나, 이에 제한되는 것은 아니다.In addition, the alpha lipoic acid can restore damaged parasympathetic nerves, and specifically can increase the expression of glial cell-derived neurotrophic factor family receptor 2 (GFR2), which is a marker of parasympathetic nerves. and may increase the expression of brain-derived neurotrophic factor (BDNF) and neurturin related to parasympathetic nerves, but is not limited thereto.
또한, 상기 알파리포산은 침샘을 재생시킬 수 있으며, 구체적으로 침샘의 재생과 관련된 헤지호그(Hedgehog, Hh) 신호전달과 관련된 소닉 헤지호그(Sonic Hedgehog, Shh) 및 그의 수용체 패치드(Patched, Ptch) 유전자의 발현을 증가시킬 수 있고, 침샘의 재생과 관련된 줄기세포 관련 마커인 Sca-1의 발현을 증가시킬 수 있으나, 이에 제한되는 것은 아니다.In addition, the alpha lipoic acid can regenerate the salivary glands, specifically hedgehog (Hedgehog, Hh) signaling related to the regeneration of the salivary gland Sonic hedgehog (Sonic Hedgehog, Shh) and its receptor Patched (Patched, Ptch) It is possible to increase the expression of the gene and may increase the expression of Sca-1, a stem cell-related marker related to regeneration of the salivary gland, but is not limited thereto.
또한, 본 발명은 알파리포산(α-Lipoic acid) 또는 이의 식품학적으로 허용되는 염을 포함하는 침분비 저하증 예방 또는 개선용 건강기능식품에 관한 것이다.In addition, the present invention relates to a health functional food for preventing or improving hyposalivation, comprising α-Lipoic acid or a pharmaceutically acceptable salt thereof.
상기 침분비 저하증은 침샘 손상으로부터 유발되는 것일 수 있고, 상기 침샘 손상은 방사성요오드 치료로 유발되는 것일 수 있으나, 이에 제한되는 것은 아니다.The hyposalivation may be caused by damage to the salivary glands, and the damage to the salivary glands may be caused by radioiodine treatment, but is not limited thereto.
상기 알파리포산의 효능, 작용 메커니즘 등은 상기 전술한 범위 내일 수 있다.The efficacy, mechanism of action, etc. of the alpha lipoic acid may be within the above-mentioned range.
본 발명에서 "식품학적으로 허용되는"이란 상기 화합물에 노출되는 세포나 인간에게 독성이 없는 특성을 나타내는 것을 의미한다.In the present invention, "food pharmaceutically acceptable" means exhibiting a property that is not toxic to cells or humans exposed to the compound.
본 발명에서 "식품학적으로 허용되는 염"이란, 본 발명에 따른 특정 화합물과 비교적 무독성인 산 또는 염기를 이용해서 조제되는 염을 의미하며, "염"에 관한 전술한 범위 내일 수 있다.In the present invention, "food pharmaceutically acceptable salt" means a salt prepared using a specific compound according to the present invention and a relatively non-toxic acid or base, and may be within the above-described range for "salt".
본 발명의 건강기능식품은 담체, 희석제, 부형제 및 첨가제 중 하나 이상을 더 포함하여 정제, 환제, 산제, 과립제, 분말제, 캡슐제 및 액제 제형으로 이루어진 군에서 선택된 하나로 제형될 수 있다. 본 발명의 추출물을 첨가할 수 있는 식품으로는, 각종 식품류, 분말, 과립, 정제, 캡슐, 시럽제, 음료, 껌, 차, 비타민 복합제, 건강기능성 식품류 등이 있다.The health functional food of the present invention may be formulated into one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid formulations, further comprising one or more of carriers, diluents, excipients and additives. Foods to which the extract of the present invention can be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gums, tea, vitamin complexes, health functional foods, and the like.
상기 본 발명에 더 포함될 수 있는 첨가제로는, 천연 탄수화물, 향미제, 영양제, 비타민, 광물(전해질), 풍미제(합성 풍미제, 천연 풍미제 등), 착색제, 충진제(치즈, 초콜렛 등), 팩트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH조절제, 안정화제, 방부제, 산화 방지제, 글리세린, 알콜, 탄산화제 및 과육으로 이루어진 군으로부터 선택된 1종 이상의 성분을 사용할 수 있다. Additives that may be further included in the present invention include natural carbohydrates, flavoring agents, nutrients, vitamins, minerals (electrolytes), flavoring agents (synthetic flavoring agents, natural flavoring agents, etc.), coloring agents, fillers (cheese, chocolate, etc.), At least one component selected from the group consisting of facic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, antioxidants, glycerin, alcohols, carbonation agents, and pulp may be used. .
상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상기 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents (taumartin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
상기 외에 본 발명의 건강기능식품은 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, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명에 따른 조성물은 천연 과일 쥬스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. In addition to the above, the health functional food of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, colorants and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the composition according to the present invention may contain the pulp for the production of natural fruit juices and vegetable beverages. These components may be used independently or in combination.
상기 담체, 부형제, 희석제 및 첨가제의 구체적인 예로는 이에 한정하는 것은 아니나, 락토즈, 덱스트로즈, 슈크로즈, 솔비톨, 만니톨, 에리스리톨, 전분, 아카시아 고무, 인산칼슘, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 미세결정성 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 폴리비닐피롤리돈, 메틸셀룰로즈, 물, 설탕시럽, 메틸셀룰로즈, 메틸 하이드록시 벤조에이트, 프로필하이드록시 벤조에이트, 활석, 스테아트산 마그네슘 및 미네랄 오일로 이루어진 그룹으로부터 선택된 1종 이상이 사용되는 것이 바람직하다.Specific examples of the carrier, excipient, diluent and additive include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium Silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and at least one selected from the group consisting of mineral oil is preferably used.
본 발명의 건강기능식품을 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다.In the case of formulating the health functional food of the present invention, it is prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, and surfactant usually used.
이하, 본 발명을 구체적으로 설명하기 위해 실시예를 들어 상세하게 설명하기로 한다.Hereinafter, examples will be given to describe the present invention in detail.
실시예Example
1. 실험 방법1. Experimental method
(1) 동물 연구(1) Animal studies
암컷 C57BL/6 마우스 (18-22 g; Orient Bio Inc., 한국 성남시)는 통제된 온도/조명 조건 하에서 물과 표준 마우스 식사를 무료로 이용할 수 있는 환경에서 유지되었다. 동물 연구는 국립 암 센터 기관 동물 윤리위원회(NCC-16-325B)에서 발행한 지침에 따라 수행되었다. 동물을 다음 4개의 그룹으로 나누었다(n = 그룹당 10 마리) : 그룹 I, 정상 대조군; 그룹 II, ALA 단독 (100 mg/kg; 부광 제약, 서울); 그룹 III, RI 단독 (0.01 mCi/g 체중 경구, 131I; New Korea Industrial, Seoul, Korea); 및 그룹 IV, RI 노출 24시간 전 및 30분 전에 ALA 처리 (ALA + RI). 우리는 이전 연구를 기반으로 ALA의 복용량과 빈도를 선택하였다.Female C57BL/6 mice (18-22 g; Orient Bio Inc., Seongnam, Korea) were maintained in an environment with free access to water and standard mouse meals under controlled temperature/light conditions. Animal studies were performed according to guidelines issued by the National Cancer Center Institutional Animal Ethics Committee (NCC-16-325B). Animals were divided into the following 4 groups (n = 10 animals per group): group I, normal control; Group II, ALA alone (100 mg/kg; Bukwang Pharmaceutical, Seoul); Group III, RI alone (0.01 mCi/g body weight oral, 131I; New Korea Industrial, Seoul, Korea); and Group IV, ALA treatment (ALA + RI) 24 hours and 30 minutes prior to RI exposure. We selected the dose and frequency of ALA based on previous studies.
(2) 침(Salivary) 기능 측정(2) Measurement of Salivary function
침 분비를 측정하여 침 기능을 평가하였다. 필로카르핀 (0.01 mL/g 체중 i.p .; Isopto Carpine; Alcon Korea Ltd., Seoul, Korea)을 주사하고 8분 후 입에서 침 배출량을 5분 동안 채취하였다. 마우스의 총 체중을 측정한 후, 신선한 튜브에 필로카르핀 주사 후 10분 동안 축적된 침의 총량과 부피를 체중으로 정규화하였다. 침 지연 시간과 유속도 측정되었습니다. 침 유속(총 침 중량을 수집 시간으로 나눈 값)과 지연 시간(자극에서 침 분비 시작까지의 시간)을 계산하였다.Salivary function was assessed by measuring salivary secretion. After injection of pilocarpine (0.01 mL/g body weight i.p.; Isopto Carpine; Alcon Korea Ltd., Seoul, Korea), saliva discharge was collected from the mouth for 5 minutes after 8 minutes. After measuring the total body weight of the mice, the total amount and volume of saliva accumulated for 10 minutes after pilocarpine injection into a fresh tube was normalized to body weight. The needle delay time and flow rate were measured. Salivary flow rate (total needle weight divided by collection time) and delay time (time from stimulation to onset of salivation) were calculated.
(3) 조직 병리학(Histopathology)(3) Histopathology
조직을 0.1M PBS의 4 % 파라포름알데히드에 고정하고 파라핀을 묻힌 후 5 ㎛로 잘랐다. 절편은 HE로 염색되었다. HE 염색에서 조직 병리학적 손상은 ×200 배율 필드에서 세포질 액포를 갖는 선포 세포의 수를 등급 화하여 점수를 매겼다. 0, 0-1; 1, 2-5; 2, 5-10; 3, 10-15; 4, 15-20; 5, > 20. 콜라겐 침착 정도를 분석하기 위해 MT(Masson 's Trichrome kit, Sigma Diagnostics, St. Louis, MO, USA)로 절편을 염색하였다. 400배 배율로 무작위로 선택된 SG(침샘) 필드를 각 마우스에서 평가하고, NIS-Elements BR 3.2 (Nikon, Japan)를 사용하여 트리크롬-양성 신호의 밀도를 분석하였다.Tissues were fixed in 4% paraformaldehyde in 0.1M PBS, embedded in paraffin, and cut into 5 μm. Sections were stained with HE. Histopathological damage in HE staining was scored by grading the number of acinar cells with cytoplasmic vacuoles in a ×200 magnification field. 0, 0-1; 1, 2-5; 2, 5-10; 3, 10-15; 4, 15-20; 5, > 20. Sections were stained with MT (Masson's Trichrome kit, Sigma Diagnostics, St. Louis, MO, USA) to analyze the degree of collagen deposition. Randomly selected SG (salivary gland) fields at 400x magnification were evaluated in each mouse, and the density of trichrome-positive signals was analyzed using NIS-Elements BR 3.2 (Nikon, Japan).
(4) 조직 및 TUNEL 분석의 형태학적 분석(4) Tissue and morphological analysis of TUNEL analysis
SG는 즉시 실온에서 4% 파라포름알데히드에 넣고, 파라핀에 묻힌 다음 5 ㎛ 두께로 절단하였다. SG는 Masson의 trichrome (MT)과 hematoxylin 및 eosin (H & E)으로 염색되었다. ApopTag Plus In Situ Apoptosis Kit (Chemicon Int., Temecula, CA, USA)를 사용하여 TUNEL 분석에 의해 턱밑샘(submaxillary gland) 조직의 세포 사멸을 결정하였다. TUNEL-양성 세포는 ×400 배율에서 검출되었고, TUNEL-양성 세포는 10개의 무작위 고출력 필드에서 계수되었다. TUNEL 분석은 RI 노출 후 30 일 및 90 일에 수행되었다. 단면은 광학 현미경으로 시각화하고 디지털 이미지를 캡처 및 분석하였다. 데이터는 10개의 랜덤 필드에서 NIS-Elements BR 3.2 (Nikon, Japan)를 사용하여 신호 강도로 분석되었으며 폴드 변화로 설명되었다. 폴드 변경은 각 그룹의 최종 값과 30일 째 대조군 그룹의 값의 비율로 계산되었다("1"로 설정).SG was immediately put in 4% paraformaldehyde at room temperature, embedded in paraffin, and then cut to a thickness of 5 μm. SGs were stained with Masson's trichrome (MT) and hematoxylin and eosin (H&E). Apoptosis of submaxillary gland tissues was determined by TUNEL analysis using the ApopTag Plus In Situ Apoptosis Kit (Chemicon Int., Temecula, CA, USA). TUNEL-positive cells were detected at ×400 magnification, and TUNEL-positive cells were counted in 10 random high-power fields. TUNEL analysis was performed at 30 and 90 days post RI exposure. Sections were visualized by light microscopy and digital images were captured and analyzed. Data were analyzed as signal intensity using NIS-Elements BR 3.2 (Nikon, Japan) in 10 random fields and accounted for by fold change. Fold change was calculated as the ratio of the final value of each group to the value of the control group at day 30 (set as "1").
(5) SA-β-Gal 염색(5) SA-β-Gal staining
조직에서 SA-β-gal을 검출하기 위해 SG를 실온에서 1x 고정 용액에 15분 동안 고정하고 1x 인산염 완충 식염수(PBS)로 두 번 세척하고 제조업체의 지침에 따라 SA-β-gal 염색 키트 (BioVision, Mountain View, CA, USA)를 사용하여 37 ℃에서 밤새 염색하였다. 염색된 조직은 청색으로 발색하는 동안 현미경으로 관찰되었다. 다음으로 광학 현미경으로 절편을 시각화하고 디지털 이미지를 캡처 및 분석하였다. 데이터는 10개의 랜덤 필드에서 NIS-Elements BR 3.2 (Nikon, Japan)를 사용하여 신호 강도로 분석되었으며 폴드 변화로 설명되었다. 폴드 변경은 각 그룹의 최종 값과 30 일째 대조군 그룹의 값의 비율로 계산되었다 ("1"로 설정).To detect SA-β-gal in tissues, SGs were fixed in 1x fixative solution at room temperature for 15 min, washed twice with 1x phosphate-buffered saline (PBS), followed by a SA-β-gal staining kit (BioVision) according to the manufacturer's instructions. , Mountain View, CA, USA) was used for staining overnight at 37 °C. The stained tissue was observed under a microscope while developing blue. The sections were then visualized under a light microscope and digital images were captured and analyzed. Data were analyzed as signal intensity using NIS-Elements BR 3.2 (Nikon, Japan) in 10 random fields and accounted for by fold change. Fold change was calculated as the ratio of the final value of each group to the value of the control group at day 30 (set as "1").
(6) 동물 연구를 위한 SPECT 프로토콜(6) SPECT protocol for animal studies
RI 후 30일 및 90일에 technetium pertechnetate (55.5 MBq, [ 99mTc]TcO4 -; New Korea Industrial)를 이소플루란(공기 중의 2 vol %)을 사용하여 전체 이미징 프로토콜 동안 무의식 상태로 유지된 마취된 마우스의 복강 내(intraperitoneally, i.p.)로 투여하였다. 전신 단-광자 방출 컴퓨터 단층 촬영(SPECT) 이미징은 [ 99mTc]TcO4 - 주사 후에 즉시 시작되었고, 120분 동안 매 10분 마다 반복되었다(NanoSPECT; Bioscan Inc., Washington, DC, USA). 전체적으로 마우스 당 13 개의 이미지를 얻었다. 그런 다음 새로운 필로카르핀 용액 (0.5 mg/mL)을 PBS에 준비하고 SPECT 60 분 후에 0.01 mL/g 체중 (i.p.)으로 투여하였다.At 30 and 90 days post RI, technetium pertechnetate (55.5 MBq, [ 99m Tc]TcO4 − ; New Korea Industrial) was administered under anesthesia maintained unconscious during the entire imaging protocol using isoflurane (2 vol % in air). The mice were administered intraperitoneally (ip). Whole-body single-photon emission computed tomography (SPECT) imaging was started immediately after [ 99m Tc]TcO4 -injection and repeated every 10 minutes for 120 minutes (NanoSPECT; Bioscan Inc., Washington, DC, USA). In total, 13 images were obtained per mouse. Then, a fresh pilocarpine solution (0.5 mg/mL) was prepared in PBS and administered at 0.01 mL/g body weight (ip) 60 minutes after SPECT.
(7) 전신 SPECT 프로토콜(7) Whole body SPECT protocol
전신 SPECT 이미지는 4개의 다중 핀홀 콜리메이터가 장착된 대형 시야각 회전 감마 카메라를 사용하여 얻었다. 획득 매개 변수는 다음과 같다 : 360°에 걸쳐 24개의 투영, 원형 궤도 및 6분의 총 획득 시간 (투영 당 4초). 반복적 재구성 알고리즘(iterative reconstruction algorithm)을 사용하여 단층 촬영 이미지를 재구성하였다.Whole-body SPECT images were obtained using a large field-of-view rotating gamma camera equipped with four multi-pinhole collimators. Acquisition parameters are: 24 projections over 360°, circular trajectory, and a total acquisition time of 6 minutes (4 seconds per projection). The tomographic images were reconstructed using an iterative reconstruction algorithm.
(8) 면역조직화학(Immunohistochemistry)(8) Immunohistochemistry
탈파라핀화한 후, 절편을 다클론성 항-AQP5 (Abcam)에 대한 1차 항체와 함께 배양한 다음, 바이오틴 결합된 2차 IgG (1:200으로 희석됨; Vector Laboratories, Burlingame, CA, USA), 아비딘-바이오틴-과산화효소 복합체(avidin-biotin-peroxidase complex) (ABC 엘리트 키트(Elite Kit); Vector Laboratories) 및 diaminobenzidine tetrahydrochloride과 배양되었다.After deparaffinization, sections were incubated with primary antibody against polyclonal anti-AQP5 (Abcam), followed by biotinylated secondary IgG (diluted 1:200; Vector Laboratories, Burlingame, CA, USA). ), avidin-biotin-peroxidase complex (ABC Elite Kit; Vector Laboratories) and diaminobenzidine tetrahydrochloride.
다음으로 우리는 광학현미경에 의해 절편을 시각화하였고, 디지털 사진을 찍고 분석하였다.Next, we visualized the sections by light microscopy, took digital pictures and analyzed them.
데이터는 10개의 랜덤 필드에서 NIS-Elements BR 3.2 (Nikon, Japan)를 사용하여 신호 강도로 분석되었으며 폴드 변화로 설명되었다. 폴드 변경은 각 그룹의 최종 값과 30 일째 대조군 그룹의 값의 비율로 계산되었다 ("1"로 설정).Data were analyzed as signal intensity using NIS-Elements BR 3.2 (Nikon, Japan) in 10 random fields and accounted for by fold change. Fold change was calculated as the ratio of the final value of each group to the value of the control group at day 30 (set as "1").
(9) 통계 분석(Statistical analysis)(9) Statistical analysis
통계학적 분석은 Graph Pad Prism 5 (Graph Pad Software Inc., La Jolla, CA, USA)을 이용해 수행되었다. Kruskal-Wallis test에 이어 Dunn's test를 사용한 post hoc testing이 두 그룹 간의 차이점을 시험하기 위해 사용되었다. P-value < 0.05는 유의미한 것으로 간주되었다.Statistical analysis was performed using Graph Pad Prism 5 (Graph Pad Software Inc., La Jolla, CA, USA). Post hoc testing using Dunn's test followed by Kruskal-Wallis test was used to test the differences between the two groups. P -value < 0.05 was considered significant.
2. 실험 결과2. Experimental results
(1) ALA는 방사성요오드 노출로 인한 체중 감소 및 손상된 침 분비를 개선한다.(1) ALA improves weight loss and impaired salivation due to radioiodine exposure.
RI 처리 마우스 그룹의 평균 체중은 90일에 대조군보다 유의하게 낮았고, RI+ALA 및 ALA 그룹의 체중은 대조군과 유사하였다(도 1A). RI+ALA 처리 그룹의 침 유속은 RI 후 30일 및 90일에 RI 단독 그룹보다 높았다(도 1B). RI 후 90일에 RI 단독 그룹에 비해 RI+ALA 그룹에서 침 지연이 향상되었다(도 1C).The average body weight of the RI-treated mouse group was significantly lower than that of the control group at 90 days, and the body weights of the RI+ALA and ALA groups were similar to those of the control group ( FIG. 1A ). The salivary flow rate of the RI+ALA-treated group was higher than that of the RI-only group at 30 and 90 days after RI (Fig. 1B). At 90 days after RI, salivation delay was improved in the RI+ALA group compared to the RI alone group (FIG. 1C).
(2) 갑상선 기능에 대한 ELISA(Enzyme-Linked Immunosorbent Assay)(2) ELISA (Enzyme-Linked Immunosorbent Assay) for Thyroid Function
RI 치료 후 ALA가 갑상선 기능에 미치는 영향을 확인하기 위해 갑상선 자극 호르몬(TSH)에 대한 ELISA를 수행하였다(30 일 및 90 일 모두). TSH 수준은 RI 단독 그룹에서 유의하게 증가했으며 ALA 처리에 의해 감소하는 경향이 있지만 유의하지 않았다 (도 2). 갑상선 기능은 RI 단독 군에 비해 RI+ALA 군에서 ALA 치료에 의해 유의한 변화가 없었기 때문에 ALA 전처리가 RI 갑상선 치료 효과를 감소시키지 않음을 알 수 있다.To determine the effect of ALA on thyroid function after RI treatment, ELISA for thyroid stimulating hormone (TSH) was performed (both 30 and 90 days). TSH levels were significantly increased in the RI alone group and tended to decrease by ALA treatment, but were not significant (Fig. 2). Thyroid function was not significantly changed by ALA treatment in the RI+ALA group compared to the RI alone group, suggesting that ALA pretreatment does not reduce the RI thyroid treatment effect.
(3) ALA는 RI 치료로 인한 구조적 변화를 감소시킨다.(3) ALA reduces structural changes due to RI treatment.
조직학적 변화는 RI 치료 후 30일 및 90일에 헤마톡실린과 에오신(HE) 및 Masson trichrome(MT) 염색에 의해 확인되었다. 대조군은 치밀한 선포와 완전히 발달된 관을 가진 잘 구분된 소엽을 보였다. RI 후 30일 및 90일에 RI 단독 그룹은 세포질 공포화의 다 초점 영역을 나타냈다. ALA+RI 그룹은 대조군과 유사한 구조를 그대로 유지하였다 (도 3A-C). 확산 섬유성 조직은 RI 후 30일 및 90일 모두에서 RI 단독 그룹에서 관찰되었다 (도 4A, 4B). 섬유증에 대한 신호는 30일보다 90일에 더 강했다. ALA+RI 그룹은 RI 단독 그룹보다 혈관주위 섬유증(perivascular fibrosis) 및 관주위 섬유증(periductal fibrosis)이 더 적게 나타났다(도 4A-C).Histological changes were confirmed by hematoxylin and eosin (HE) and Masson trichrome (MT) staining at 30 and 90 days after RI treatment. The control group showed well-separated lobules with dense acinar and fully developed ducts. At 30 and 90 days post-RI, the RI-only group exhibited multifocal areas of cytoplasmic vacuolization. The ALA+RI group maintained a structure similar to that of the control group (FIGS. 3A-C). Diffuse fibrous tissue was observed in the RI-only group at both 30 and 90 days post-RI (Figs. 4A, 4B). The signal for fibrosis was stronger at 90 days than at 30 days. The ALA+RI group showed fewer perivascular fibrosis and periductal fibrosis than the RI alone group (FIGS. 4A-C).
(4) ALA는 RI-유도 침 세포 사멸을 감소시킨다.(4) ALA reduces RI-induced apoptosis.
RI-유도 침 세포 사멸에 대한 ALA의 효과를 결정하기 위해 말단 데옥시뉴클레오티딜 transferase biotin-dUTP nick end labeling (TUNEL) 분석이 수행되었다. 희미한 신호가 RI 치료 후 30 일 및 90 일에 대조군과 ALA 단독 그룹 모두에서 발견되었으며; 이들은 도관 세포보다 선포에서 더 풍부했으며 90일보다 30일에 더 뚜렷하였다(도 5A, 5B). 그러나 양성 신호는 대조군 및 ALA 단독 그룹에 비해 RI 단독 그룹에서 유의하게 증가하였다. 더욱이, 총 사멸 세포 수는 RI 치료 후 30일 및 90일 모두에서 ALA+RI 그룹에서 RI 단독 그룹에 비해 현저히 낮았다(도 5C).Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assay was performed to determine the effect of ALA on RI-induced apoptosis. A faint signal was found in both the control and ALA-only groups at 30 and 90 days after RI treatment; They were more abundant in acinar than ductal cells and more pronounced at 30 days than at 90 days (Figures 5A, 5B). However, the positive signal was significantly increased in the RI-only group compared to the control and ALA-only groups. Moreover, the total number of apoptotic cells was significantly lower in the ALA+RI group compared to the RI alone group at both 30 and 90 days after RI treatment (Fig. 5C).
(5) ALA는 SG에서 RI-유도 세포 노화를 방지한다.(5) ALA prevents RI-induced cellular senescence in SGs.
RI-유도 SG에서 세포 노화를 조사하기 위해 노화의 바이오 마커인 senescence-associated β-galactosidase(SA-β-gal)를 위해 SG를 염색하였다. RI 단독 그룹에서 선포 및 관 세포는 SA-β-gal에 대해 유의하게 양성이었으며, 염색은 30일 보다 90일에 뚜렷하게 더 컸다(도 6A, 6B). ALA+RI 그룹은 세포 노화에 대한 양성 신호의 현저한 감소를 보여주었다(도 6C)To investigate cellular senescence in RI-induced SGs, SGs were stained for senescence-associated β-galactosidase (SA-β-gal), a biomarker of senescence. In the RI-only group, acinar and duct cells were significantly positive for SA-β-gal, and staining was significantly greater at 90 days than at 30 days ( FIGS. 6A, 6B ). The ALA+RI group showed a significant decrease in the positive signal for cellular senescence (Fig. 6C).
(6) ALA는 방사성요오드 유발 AQP5 발현 감소를 회복시킨다.(6) ALA restores the radioiodine-induced decrease in AQP5 expression.
침 상피세포 마커인 아쿠아포린 5(AQP-5)에 대한 염색은 대조군에서 선포가 풍부한 SG를 나타냈다. 대조적으로, RI 단독 그룹은 RI 후 30일 및 90일 모두에서 몇 개의 AQP-5 양성 영역을 보여주었다(도 7A, 7B). ALA+RI 그룹은 정상 대조군과 유사한 염색을 나타냈다(도 7C).Staining for aquaporin 5 (AQP-5), a salivary epithelial cell marker, revealed acinar-rich SGs in the control group. In contrast, the RI-only group showed several AQP-5 positive regions at both 30 and 90 days post-RI (FIGS. 7A, 7B). The ALA+RI group showed staining similar to that of the normal control group ( FIG. 7C ).
(7) ALA는 RI-유도 침 기능 장애를 개선한다.(7) ALA ameliorates RI-induced acupuncture dysfunction.
SPECT(single-photon emission computed tomography)를 사용하여 RI 치료 후 침 기능을 검사하였다. RI 후 30일 째의 SPECT 이미지에 따르면 99mTc pertechnetate 제거량은 다른 그룹에 비해 RI 단독 그룹에서 더 낮았다. 99mTc pertechnetate 제거 정도는 110분 및 120분에 RI 단독 그룹에서 훨씬 더 낮았다. ALA 단독 그룹에서 99mTc pertechnetate의 배출은 대조군과 유사하였다(도 8A, 8B). RI 후 90일 째에 99mTc pertechnetate 제거 수준은 70, 90, 100, 110 및 120 분에 다른 그룹에 비해 RI 단독 그룹에서 현저히 낮았다. ALA 단독 그룹에서 99mTc pertechnetate의 배출은 대조군에서보다 더 나았다(도 9A, 9B). 이러한 관찰은 ALA가 RI 치료 후 침 기능 장애를 개선했음을 나타낸다.Acupuncture function was examined after RI treatment using single-photon emission computed tomography (SPECT). According to the SPECT image on the 30th day after RI, the amount of 99m Tc pertechnetate removed was lower in the RI-only group than in the other groups. The degree of 99m Tc pertechnetate clearance was much lower in the RI alone group at 110 and 120 minutes. The excretion of 99m Tc pertechnetate in the ALA-only group was similar to that of the control group ( FIGS. 8A and 8B ). At 90 days after RI, the level of 99m Tc pertechnetate clearance was significantly lower in the RI-only group compared to the other groups at 70, 90, 100, 110 and 120 min. The excretion of 99m Tc pertechnetate in the ALA-only group was better than in the control group ( FIGS. 9A and 9B ). These observations indicate that ALA improved acupuncture dysfunction after RI treatment.

Claims (6)

  1. 알파리포산(α-Lipoic acid) 또는 이의 약학적으로 허용되는 염을 포함하는 침분비 저하증 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating hyposalivation, comprising alpha-lipoic acid or a pharmaceutically acceptable salt thereof.
  2. 청구항 1에 있어서, 상기 침분비 저하증은 침샘 손상으로부터 유발되는 것인, 약학적 조성물.The pharmaceutical composition of claim 1, wherein the hyposalivation is caused by damage to the salivary glands.
  3. 청구항 2에 있어서, 상기 침샘 손상은 방사성요오드 치료로 유발되는 것인, 약학적 조성물.The pharmaceutical composition of claim 2, wherein the salivary gland damage is caused by radioiodine treatment.
  4. 알파리포산(α-Lipoic acid) 또는 이의 식품학적으로 허용되는 염을 포함하는 침분비 저하증 예방 또는 개선용 건강기능식품.A health functional food for preventing or improving hyposalivation, comprising alpha-Lipoic acid or a pharmaceutically acceptable salt thereof.
  5. 청구항 4에 있어서, 상기 침분비 저하증은 침샘 손상으로부터 유발되는 것인, 건강기능식품.The health functional food according to claim 4, wherein the hyposalivation is caused by damage to the salivary glands.
  6. 청구항 5에 있어서, 상기 침샘 손상은 방사성요오드 치료로 유발되는 것인, 건강기능식품.The health functional food according to claim 5, wherein the salivary gland damage is caused by radioiodine treatment.
PCT/KR2020/014087 2020-10-15 2020-10-15 Pharmaceutical composition and health functional food for preventing or treating hyposalivation WO2022080529A1 (en)

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US4906455A (en) * 1988-04-15 1990-03-06 Wm. Wrigley Jr. Company Method for treating xerostomia employing chewing gum containing relatively insoluble, hydrophobic, food-grade organic acid
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KR20200083370A (en) * 2018-12-28 2020-07-08 경상대학교병원 Pharmaceutical composition and health functional food for prevention or treatment of salivation
KR20200118128A (en) * 2018-02-05 2020-10-14 셀릭스 바이오 프라이빗 리미티드 Combination of antimuscarinic or anticholinergic and lipoic acid and uses thereof

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US4906455A (en) * 1988-04-15 1990-03-06 Wm. Wrigley Jr. Company Method for treating xerostomia employing chewing gum containing relatively insoluble, hydrophobic, food-grade organic acid
KR20120092152A (en) * 2009-11-12 2012-08-20 아카시아 파마 리미티드 Use of bethanechol for treatment of xerostomia
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