WO2020246671A1 - Composition for promoting bone growth including carrot leaf as active ingredient - Google Patents

Composition for promoting bone growth including carrot leaf as active ingredient Download PDF

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Publication number
WO2020246671A1
WO2020246671A1 PCT/KR2019/016384 KR2019016384W WO2020246671A1 WO 2020246671 A1 WO2020246671 A1 WO 2020246671A1 KR 2019016384 W KR2019016384 W KR 2019016384W WO 2020246671 A1 WO2020246671 A1 WO 2020246671A1
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growth
short stature
bone growth
promoting bone
carrot leaf
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PCT/KR2019/016384
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French (fr)
Korean (ko)
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고병섭
김혜진
육진아
양현
황주태
정동호
편보정
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한국한의학연구원
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Publication of WO2020246671A1 publication Critical patent/WO2020246671A1/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
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • 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
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • 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
    • A23V2200/306—Foods, ingredients or supplements having a functional effect on health having an effect on bone mass, e.g. osteoporosis prevention
    • 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
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts
    • 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
    • A23V2300/00—Processes
    • A23V2300/14—Extraction

Definitions

  • Bone is a tissue composed of calcium and phosphorus, and its main components are calcium phosphate double salt, collagen fibers, glycoprotein, and proteoglycan, and are organs that make up and support the body. It protects internal organs, supports muscles so that they can move, and also plays a role in forming and maintaining the shape of a person, but bone marrow in bones can produce blood, which is the body's oxygen carrier.
  • Osteoblasts one of the types of bone cells, secrete collagen and alkaline phosphatase to form bone.Although it is not yet known what function of alkaline phosphatase it performs, bone cell production is high. It is known to have high levels in the plasma of patients. In addition, osteoclasts are cells that destroy bones and play a role in dissolving bones and releasing phosphate and calcium into plasma.
  • Calcium and phosphorus the main elements of bone, exist as a complex of two ions when forming bone, but exist in an ionized form in serum when moving. Depending on the concentration of calcium ions in the blood, homeostasis can be maintained as calcium reabsorption is increased or excreted.
  • An object of the present invention is to improve bone growth through growth plate expansion when administered or ingested to children or adolescents in the growing period, and to increase bone density and improve mineral concentration in bone when administered or ingested to adults with low bone density. It is to provide a composition for promoting bone growth comprising a carrot leaf extract.
  • a food composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
  • the carrot leaf extract may be extracted with water, alcohol having 1-4 carbon atoms, or a mixed solvent thereof.
  • the food composition for promoting bone growth may be one to increase the activity of at least one bone cell differentiation gene selected from runx2, osteocalcin, and colla1.
  • the food composition for promoting bone growth includes familial short stature, constitutional growth delay, osteocartilage dysplasia, short stature due to chromosomal abnormalities, short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, short stature due to Noonan syndrome, It may be for improving any one symptom selected from short stature due to chronic systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, and short stature due to Cushing's syndrome.
  • the food composition for promoting bone growth may be formulated as one or more selected from granules, powders, tablets, coated tablets, capsules, liquids, syrups, juices, suspensions, emulsions and drops, and may be added to food.
  • a health functional food composition for promoting bone growth comprising carrot leaf extract as an active ingredient is provided.
  • composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
  • the carrot leaf extract may be extracted with water, alcohol having 1-4 carbon atoms, or a mixed solvent thereof.
  • the pharmaceutical composition for promoting bone growth may increase the activity of one or more bone cell differentiation genes selected from runx2, osteocalcin, and colla1.
  • the pharmaceutical composition for promoting bone growth includes familial short stature, constitutional growth delay, osteocartilage dysplasia, short stature due to chromosomal abnormalities, short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, short stature due to Noonan syndrome, It may be for the treatment of any one symptom selected from short stature due to chronic systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, and short stature due to Cushing's syndrome.
  • the food composition for promoting bone growth may be formulated as one or more selected from granules, powders, tablets, coated tablets, capsules, liquids, syrups, juices, suspensions, emulsions and drops.
  • a novel use of carrot leaf extract for manufacturing a medicament for promoting bone growth is provided.
  • the medicament for promoting bone growth may be a medicament for preventing or treating growth disorders.
  • a method for treating a growth disorder comprising administering a pharmaceutical composition comprising a carrot leaf extract as an active ingredient to a patient with a growth disorder.
  • composition for promoting bone growth comprising the carrot leaf extract of the present invention improves bone growth through growth plate expansion when administered or ingested to children or adolescents in the growing period, and increases bone density when administered or ingested to adults with low bone density. It can increase and improve the mineral concentration in the bone. In addition, since it promotes bone growth, it can be used for prevention or treatment of growth disorders.
  • Figure 8 shows the results of measuring the growth plate thickness in the tibia according to Experimental Example 7.
  • Carrot leaves 500 g were extracted with reflux cooling twice for 3 hours each with 10 times of 70% ethanol, and the filtered solution was concentrated using a vacuum concentrator and then freeze-dried to prepare a carrot leaf extract.
  • MC3T3-E1 cells a precursor bone cell line
  • MC3T3-E1 cells were cultured in a 5% CO 2 , 37°C incubator using an alpha-MEM nutrient medium containing 5% FBS.
  • Differentiation into bone cells was conducted in a differentiation medium containing 100 nM dexamethasone, 10 mM ⁇ -glycerophosphate, and 50 ⁇ g/ml ascorbic acid in cells before passage 10.
  • Media, CM to induce differentiation for 7, 14 and 21 days.
  • the carrot leaf extract was treated in a differentiation medium (DM) at concentrations of 10, 50, and 100 ⁇ g/ml, respectively, and cultured for 7, 14, and 21 days.
  • Bone differentiation activity by carrot leaves was evaluated by measuring the expression level of degrading enzyme, the degree of calcium salt deposition (Alizarin Red staining), and expression of differentiation-related genes.
  • CM differentiation medium
  • Carrot leaf extract 100 ⁇ g / ml for 7 days 5% CO 2 was cultured in a 37 °C incubator. After a certain cultivation time, the medium was collected and used for analysis. For analysis, Alkaline Phosphatase Assay Kit (Abcam, ab83369) was used, and absorbance was measured at 405 nm using a multiplate reader, and the result of measuring the ALP expression level by the carrot leaf extract is shown in FIG. 2.
  • ALP activity was increased in Conditioned media (CM) compared to normal media (NM), and in the group (D) treated with carrot leaf extract, a higher amount of ALP than the differentiation medium Increased activity was confirmed.
  • CM Conditioned media
  • NM normal media
  • D group treated with carrot leaf extract
  • ALP shows the degree of destruction of cells, but in cell experiments, new cells are generated only when osteoblasts are destroyed, so that increasing ALP activity increases cell growth.
  • RNA extraction kit Qiagen
  • a cDNA synthesis kit Bio-Rad was used to mix with 1 ⁇ g of RNA sample, and then cDNA was synthesized using a thermal cycler (Bio-rad).
  • the obtained cDNA 1 ⁇ l and 10 pmole of primer were mixed with sybr green master mix (Thermo Fisher), and real-time PCR was performed using a thermal cycler (Thermo Fisher).
  • the base sequence of the primers for each specific gene is as follows. It was repeated three times for each experimental group and plotted using a statistical program (Prism), and the results are shown in FIG. 5.
  • the primer information is summarized in Table 1 below.
  • Control is a control group that has not been treated with any differentiation medium (DM), and 10, 50, and 100 ⁇ g/ml are differentiation medium and carrot leaf treatment group for each concentration.
  • DM differentiation medium
  • the expression of mRNA of the bone cell differentiation genes runx2, osteocalcin, and colla1 was significantly increased in the treatment period (7, 14, 21 days) and treatment concentration-dependently.
  • a 4-week-old mouse was worn in a SPF (specific pathogen free) grade breeding room where temperature and humidity were controlled, and then acclimatized for a week.
  • Five-week-old mice purified for one week were fed a diet deficient in vitamin D and calcium for 8 weeks to induce a decrease in bone growth, and the carrot leaf extract prepared according to Preparation Example 1 was administered orally for the same period of time. Efficacy for improving inhibition of bone growth was confirmed.
  • by measuring the weight and the amount of diet twice a week it was confirmed that the health status of the model and the intake of diets deficient in vitamin D and calcium were properly progressed.
  • composition of the experimental group is a normal control group (Vehicle, no treatment group), a positive control group (positive, bone growth inhibition induction and Somatotrophin 0.1 mg/kg treatment group), carrot leaf low concentration treatment group (KIOM_D_L, bone growth inhibition induction and carrot leaf 150 mg/kg treatment group), carrot leaf high concentration treatment group (KIOM_D_H, bone growth inhibition induction and carrot leaf 450 mg/kg treatment group).
  • bone density, bone mineral concentration, and growth plate thickness were measured using the deformed femur and tibia.
  • the bone density and bone mineral concentration of the femur used in the analysis were fixed in 10% neutral formalin for 3 days after an animal autopsy, and then muscle and vascular tissues other than the bone were removed, followed by Dual-energy X-ray absorptiometry (DEXA; Norland pDEXA). Saber; WI, USA), and then analyzed using Saber Research (v3.6) software, and the results of measuring bone density and mineral concentration in the femur by carrot leaf extract are shown in FIG. 7.
  • composition of the experimental group is a normal control group (Vehicle, no treatment group), a negative control group (Negative, bone growth inhibition induction group), a positive control (bone growth inhibition induction and Somatotrophin 0.1 mg/kg treatment group), carrot leaf low concentration treatment group (KIOM_D_L, bone growth inhibition induction and carrot leaf 150 mg/kg treatment group), carrot leaf high concentration treatment group (KIOM_D_H, bone growth inhibition induction and carrot leaf 450 mg/kg treatment group).
  • the thickness of the growth plate in the tibia was determined by using the above method and pretreatment, followed by demineralization, embedding, and thinning to make 5 ⁇ m paraffin sections, and observe the growth plate thickness using hematoxylin & eosin stains.
  • the thickness of the growth plate was measured using the software cellSens standard (Olympus), and the results are shown in FIG. 8.
  • composition of the experimental group is a normal control group (Vehicle, no treatment group), a negative control group (Negative, bone growth inhibition induction group), a positive control (bone growth inhibition induction and Somatotrophin 0.1 mg/kg treatment group), carrot leaf high concentration treatment group (KIOM_D_H, bone growth inhibition induction and carrot leaf 450 mg/kg treatment group).
  • composition for promoting bone growth of the present invention contains carrot leaf extract as an active ingredient.
  • the carrot leaves are mixed with an extraction solvent in a weight ratio of 1: 5 to 25, preferably 1: 8 to 15, at 40 to 70°C, preferably 50 to 60°C for 10 to 20 hours, preferably 15 to 18 After extraction for a period of time, concentration under reduced pressure is performed to prepare an extract.
  • concentration under reduced pressure is performed to prepare an extract.
  • the active ingredient of carrot leaf may be extracted in a small amount in the extract.
  • the extraction solvent for extracting each extract is water, a lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixed solvent thereof.
  • the lower alcohol may include 20 to 80% of methanol, ethanol, butanol or propanol.
  • the extraction solvent is not particularly limited, but the extract extracted with an aqueous solution of 60 to 80% ethanol preferably acts on promoting bone growth.
  • carrot leaf extract used in referring to carrot leaves in the present specification includes not only crude extract obtained by treatment with an extraction solvent, but also processed products of carrot leaf extract.
  • the carrot leaf extract may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze drying or spray drying.
  • the carrot leaf extract of the present invention has a meaning to include a carrot leaf processed product, for example, carrot leaf powder, formulated so that carrot leaf can be administered to animals.
  • the term "including as an active ingredient” means including an amount sufficient to achieve the efficacy or activity of the carrot leaf extract.
  • the carrot leaf extract is used in a concentration of 10 to 1500 ⁇ g/ml, preferably 100 to 1000 ⁇ g/ml. Since carrot leaf extract is a natural product and does not have side effects on the human body even if it is administered in excess, the upper limit of the quantity of the carrot leaf extract contained in the composition of the present invention can be selected and carried out by a person skilled in the art within an appropriate range.
  • the present invention provides a food composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
  • the growth disorder may be normal variant short stature or short stature secondary to disease.
  • the normal variant short stature may be familial short stature, delayed constitutional growth, or negotiated idiopathic short stature.
  • short kidney disease secondary to the disease may be a primary growth disorder (endogenous disorder) or a secondary growth disorder (exogenous growth disorder), and the primary growth disorder includes osteochondral dysplasia, chromosomal abnormalities (Down syndrome or Turner syndrome), unfair lightweight infants (delayed growth in the uterus), short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, and short stature due to Noonan syndrome, and the secondary growth disorders are chronic There are short stature due to systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, short stature due to Cushing's syndrome, and psychosocial dwarfism.
  • the food composition according to the present invention may be formulated in the same manner as the following pharmaceutical composition to be used as a functional food or added to various foods.
  • Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, health supplements. Etc.
  • the food composition of the present invention may include not only carrot leaf extract as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, flavoring agents and flavoring agents.
  • examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • flavoring agents natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.
  • natural flavoring agents eg, rebaudioside A, glycyrrhizin, etc.
  • synthetic flavoring agents sacharin, aspartame, etc.
  • the present invention provides a health functional food comprising a food composition for promoting bone growth comprising the carrot leaf extract as an active ingredient.
  • Health functional foods are foods prepared by adding carrot leaf extract to food ingredients such as beverages, teas, spices, chewing gums, confectionery, or encapsulating, powdering, and suspending.
  • food ingredients such as beverages, teas, spices, chewing gums, confectionery, or encapsulating, powdering, and suspending.
  • the health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis.
  • the amount of carrot leaf extract added in such health functional foods cannot be uniformly regulated depending on the type of health functional food to be targeted, but can be added within the range that does not damage the original taste of the food.
  • the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.
  • the present invention provides a pharmaceutical composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
  • the carrot leaf extract promotes bone growth, it can be used for preventing or treating growth disorders.
  • the growth disorder may be normal variant short stature or short stature secondary to disease.
  • the normal variant short stature may be familial short stature, delayed constitutional growth, or negotiated idiopathic short stature.
  • short kidney disease secondary to the disease may be a primary growth disorder (endogenous disorder) or a secondary growth disorder (exogenous growth disorder), and the primary growth disorder includes osteochondral dysplasia, chromosomal abnormalities (Down syndrome or Turner syndrome), unfair lightweight infants (delayed growth in the uterus), short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, and short stature due to Noonan syndrome, and the secondary growth disorders are chronic There are short stature due to systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, short stature due to Cushing's syndrome, and psychosocial dwarfism.
  • the pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, lubricants. Agents or flavoring agents can be used.
  • the pharmaceutical composition may include one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.
  • the formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions.
  • the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.
  • suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture.
  • Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like.
  • Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.
  • compositions formulated as liquid solutions, sterilization and biocompatible saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary.
  • diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.
  • the pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, and the like, preferably oral administration.
  • a suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, and is usually As a result, a skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prevention.
  • the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.
  • the pharmaceutical composition of the present invention may be prepared in a unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient, or may be prepared by placing it in a multi-dose container.
  • the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.
  • the present invention provides a use of a carrot leaf extract for the manufacture of a medicine or food for promoting bone growth.
  • the carrot leaf extract can be used for the treatment or improvement of diseases associated with promoting bone growth and reducing bone growth.
  • the present invention provides a method for improving, preventing or treating sleep, a disorder or insomnia comprising administering an effective amount of carrot leaf extract to a mammal.
  • mammal refers to a mammal that is the subject of treatment, observation or experimentation, and preferably refers to a human.
  • an effective amount refers to an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, which is considered by a researcher, veterinarian, doctor or other clinician, which Or an amount that induces relief of symptoms of the disorder.
  • the effective amount and the number of administrations for the active ingredient of the present invention may vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the amount of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient.
  • the extract in the case of an adult, it is preferable to administer the extract at a dose of 0.001 g/kg to 10 g/kg when administered once to several times a day.
  • the composition containing the carrot leaf extract as an active ingredient is administered in a conventional manner through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, or intradermal routes. can do.
  • composition for promoting bone growth comprising the carrot leaf extract of the present invention improves bone growth through growth plate expansion when administered or ingested to children or adolescents in the growing period, and increases bone density when administered or ingested to adults with low bone density. It can increase and improve the mineral concentration in the bone.

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Abstract

The present invention relates to a composition for promoting bone growth comprising a carrot leaf extract as an active ingredient. Accordingly, a composition for promoting bone growth comprising a carrot leaf extract according to the present invention not only has the effect of expanding the growth plate and thereby promoting bone grown when administered as medicine, food, etc. to children or adolescents who are in the growth period, but also has the effect of increasing bone density and the concentration of minerals in the bones when administered to adults having low bone density.

Description

당근잎을 유효성분으로 포함하는 뼈 성장 촉진용 조성물Composition for promoting bone growth containing carrot leaves as an active ingredient
본 발명은 당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 조성물에 관한 것으로, 더욱 상세하게는 뼈성장을 촉진시키거나 골밀도를 증가시킬 수 있는 뼈 성장 촉진용 식품 또는 약학 조성물에 관한 것이다.The present invention relates to a composition for promoting bone growth comprising a carrot leaf extract as an active ingredient, and more particularly, to a food or pharmaceutical composition for promoting bone growth capable of promoting bone growth or increasing bone density.
뼈는 칼슘과 인으로 이루어진 조직으로 인산칼슘복염, 콜라겐 섬유, glycoprotein, proteoglycan 이 주 구성물이며, 몸을 구성하고 지탱하는 역할을 수행하는 기관이다. 내부의 장기를 보호하는 역할을 수행하고 근육이 움직일 수 있도록 지탱하는 역할을 하며, 사람이 형태를 구성하고 유지하는 역할도 수행하지만, 뼈 속 골수에서는 몸의 산소운반체인 혈액을 생성할 수 있다. Bone is a tissue composed of calcium and phosphorus, and its main components are calcium phosphate double salt, collagen fibers, glycoprotein, and proteoglycan, and are organs that make up and support the body. It protects internal organs, supports muscles so that they can move, and also plays a role in forming and maintaining the shape of a person, but bone marrow in bones can produce blood, which is the body's oxygen carrier.
뼈세포의 유형 중 하나인 조골 세포는 콜라겐과 알칼리인산분해효소를 분비하여 뼈를 형성하는 역할을 수행하는데, 알칼리인산분해효소의 경우 어떠한 기능을 수행하는지에 대해 아직 알려져 있지 않으나 뼈세포 생성이 높은 환자들의 혈장 내에서 높은 수치를 나타내는 것으로 알려져 있다. 또한, 파골세포는 뼈를 파괴하는 세포로 뼈를 분해하여 인산과 칼슘을 혈장으로 흘려 보내는 역할을 수행한다. Osteoblasts, one of the types of bone cells, secrete collagen and alkaline phosphatase to form bone.Although it is not yet known what function of alkaline phosphatase it performs, bone cell production is high. It is known to have high levels in the plasma of patients. In addition, osteoclasts are cells that destroy bones and play a role in dissolving bones and releasing phosphate and calcium into plasma.
골 성장은 파골세포와 조골세포의 상호작용에 의해 이루어지는데, 파골세포에 의해 골이 흡수되면 조골세포는 다시 칼슘과 인을 조합하면서 골 성장이 이루어질 수 있다. 조골세포의 활성이 파골세포보다 높을 경우 골 성장이 이루어지며, 반대의 경우에는 골 파괴가 이루어져 골성장을 어렵게 할 수 있다.Bone growth is achieved by the interaction of osteoclasts and osteoblasts. When bone is absorbed by the osteoclasts, the osteoblasts can grow bone by combining calcium and phosphorus again. When the activity of osteoblasts is higher than that of osteoclasts, bone growth occurs, and in the opposite case, bone destruction occurs, which may make bone growth difficult.
뼈를 구성하는 주요소인 칼슘과 인은 뼈를 구성할 때는 두 이온이 복합체를 이루어 존재하지만 이동할 때는 혈청에서 이온화된 형태로 존재한다. 혈중 칼슘이온의 농도에 따라서도 칼슘 재흡수가 증가되거나 배설이 되면서 항상성이 유지될 수 있다.Calcium and phosphorus, the main elements of bone, exist as a complex of two ions when forming bone, but exist in an ionized form in serum when moving. Depending on the concentration of calcium ions in the blood, homeostasis can be maintained as calcium reabsorption is increased or excreted.
이와 같이 뼈 성장 촉진 개선에 효과적이면서도 인체에 유해하지 않고 장기간 복용하여 부작용이 거의 없는 천연 성분의 뼈 성장 촉진 개선에 유용한 물질의 개발이 필요한 실정이다.As described above, there is a need to develop a substance useful for promoting bone growth of natural ingredients that are effective in improving bone growth promotion but not harmful to the human body and have little side effects after long-term use.
본 발명의 목적은 성장기에 있는 어린이나 청소년에 투여 또는 섭취시킬 경우 성장판 확장을 통해 뼈 성장을 향상시키고, 골밀도가 낮은 성인에 투여 또는 섭취시킬 경우에는 골밀도를 높이고 골 중 미네랄 농도를 향상시킬 수 있는 당근잎 추출물을 포함하는 뼈 성장 촉진용 조성물을 제공하는 데 있다.An object of the present invention is to improve bone growth through growth plate expansion when administered or ingested to children or adolescents in the growing period, and to increase bone density and improve mineral concentration in bone when administered or ingested to adults with low bone density. It is to provide a composition for promoting bone growth comprising a carrot leaf extract.
본 발명의 하나의 측면에 따르면,According to one aspect of the invention,
당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 식품 조성물이 제공된다.There is provided a food composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
상기 당근잎 추출물은 물, 탄소수 1-4의 알코올, 또는 이들의 혼합용매에 의해 추출된 것일 수 있다.The carrot leaf extract may be extracted with water, alcohol having 1-4 carbon atoms, or a mixed solvent thereof.
상기 뼈 성장 촉진용 식품 조성물은 runx2, osteocalcin, 및 colla1 중에서 선택된 1종 이상의 골세포 분화 유전자의 활성을 증가시키는 것일 수 있다.The food composition for promoting bone growth may be one to increase the activity of at least one bone cell differentiation gene selected from runx2, osteocalcin, and colla1.
상기 뼈 성장 촉진용 식품 조성물은 가족성 저신장, 체질적 성장 지연, 골연골 이형성, 염색체 이상에 의한 저신장, 프레더-윌리 증후군에 의한 저신장, 러셀-실버 증후군에 의한 저신장, 누난 증후군에 의한 저신장, 만성 전신성 질환에 의한 저신장, 성장호르몬 결핍증에 의한 저신장, 갑상샘 저하증에 의한 저신장, 및 쿠싱증후군에 의한 저신장 중에서 선택된 어느 하나의 증상 개선을 위한 것일 수 있다.The food composition for promoting bone growth includes familial short stature, constitutional growth delay, osteocartilage dysplasia, short stature due to chromosomal abnormalities, short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, short stature due to Noonan syndrome, It may be for improving any one symptom selected from short stature due to chronic systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, and short stature due to Cushing's syndrome.
상기 뼈 성장 촉진용 식품 조성물은 과립제, 산제, 정제, 피복정, 캡슐제, 액제, 시럽, 즙, 현탁제, 유제 및 점적제 중에서 선택된 1종 이상으로 제제화되어 식품에 첨가될 수 있다.The food composition for promoting bone growth may be formulated as one or more selected from granules, powders, tablets, coated tablets, capsules, liquids, syrups, juices, suspensions, emulsions and drops, and may be added to food.
본 발명의 다른 하나의 측면에 따르면,According to another aspect of the present invention,
당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 건강기능식품 조성물이 제공된다.A health functional food composition for promoting bone growth comprising carrot leaf extract as an active ingredient is provided.
본 발명의 다른 또 하나의 측면에 따르면,According to another aspect of the present invention,
당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 약학 조성물이 제공된다.There is provided a pharmaceutical composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
상기 당근잎 추출물은 물, 탄소수 1-4의 알코올, 또는 이들의 혼합용매에 의해 추출된 것일 수 있다.The carrot leaf extract may be extracted with water, alcohol having 1-4 carbon atoms, or a mixed solvent thereof.
상기 뼈 성장 촉진용 약학 조성물은 runx2, osteocalcin, 및 colla1 중에서 선택된 1종 이상의 골세포 분화 유전자의 활성을 증가시킬 수 있다.The pharmaceutical composition for promoting bone growth may increase the activity of one or more bone cell differentiation genes selected from runx2, osteocalcin, and colla1.
상기 뼈 성장 촉진용 약학 조성물은 가족성 저신장, 체질적 성장 지연, 골연골 이형성, 염색체 이상에 의한 저신장, 프레더-윌리 증후군에 의한 저신장, 러셀-실버 증후군에 의한 저신장, 누난 증후군에 의한 저신장, 만성 전신성 질환에 의한 저신장, 성장호르몬 결핍증에 의한 저신장, 갑상샘 저하증에 의한 저신장, 및 쿠싱증후군에 의한 저신장 중에서 선택된 어느 하나의 증상 치료를 위한 것일 수 있다.The pharmaceutical composition for promoting bone growth includes familial short stature, constitutional growth delay, osteocartilage dysplasia, short stature due to chromosomal abnormalities, short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, short stature due to Noonan syndrome, It may be for the treatment of any one symptom selected from short stature due to chronic systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, and short stature due to Cushing's syndrome.
상기 뼈 성장 촉진용 식품 조성물은 과립제, 산제, 정제, 피복정, 캡슐제, 액제, 시럽, 즙, 현탁제, 유제 및 점적제 중에서 선택된 1종 이상으로 제제화될 수 있다.The food composition for promoting bone growth may be formulated as one or more selected from granules, powders, tablets, coated tablets, capsules, liquids, syrups, juices, suspensions, emulsions and drops.
본 발명의 다른 또 하나의 측면에 따르면,According to another aspect of the present invention,
뼈 성장 촉진용 의약 제조를 위한 당근잎 추출물의 신규 용도가 제공된다. A novel use of carrot leaf extract for manufacturing a medicament for promoting bone growth is provided.
상기 뼈 성장 촉진용 의약은 성장 장애의 예방 또는 치료용 의약일 수 있다.The medicament for promoting bone growth may be a medicament for preventing or treating growth disorders.
본 발명의 다른 또 하나의 측면에 따르면,According to another aspect of the present invention,
성장 장애 환자에게 당근잎 추출물을 유효성분으로 포함하는 약학 조성물을 투여하는 것을 포함하는 성장 장애의 치료방법이 제공된다.There is provided a method for treating a growth disorder comprising administering a pharmaceutical composition comprising a carrot leaf extract as an active ingredient to a patient with a growth disorder.
본 발명의 당근잎 추출물을 포함하는 뼈 성장 촉진용 조성물은 성장기에 있는 어린이나 청소년에 투여 또는 섭취시킬 경우 성장판 확장을 통해 뼈 성장을 향상시키고, 골밀도가 낮은 성인에 투여 또는 섭취시킬 경우에는 골밀도를 높이고 골 중 미네랄 농도를 향상시킬 수 있다. 또한 뼈 성장을 촉진하므로 성장 장애의 예방 또는 치료 용도로 사용될 수 있다.The composition for promoting bone growth comprising the carrot leaf extract of the present invention improves bone growth through growth plate expansion when administered or ingested to children or adolescents in the growing period, and increases bone density when administered or ingested to adults with low bone density. It can increase and improve the mineral concentration in the bone. In addition, since it promotes bone growth, it can be used for prevention or treatment of growth disorders.
도 1은 실험예 1에 따른 세포독성 효과 분석 결과이다.1 is a cytotoxic effect analysis result according to Experimental Example 1.
도 2는 실험예 2의 염기성 인산분해효소(alkaline phosphatase, ALP) 발현량 측정 결과를 나타낸 것이다.2 shows the result of measuring the expression level of alkaline phosphatase (ALP) in Experimental Example 2.
도 3은 실험예 3의 칼슘 침착 평가에 따른 염색양상을 나타낸 것이다. 3 shows the staining pattern according to the evaluation of calcium deposition in Experimental Example 3.
도 4는 실험예 3의 칼슘 침착 평가에 따른 흡광도 측정결과를 나타낸 것이다.4 shows the absorbance measurement results according to the evaluation of calcium deposition in Experimental Example 3.
도 5는 실험예 4에 따른 골세포 분화 관련 유전자의 발현 평가 결과를 나타낸 것이다.5 shows the results of evaluation of the expression of genes related to bone cell differentiation according to Experimental Example 4.
도 6은 실험예 5의 혈중 골 지표 인자 함량 변화 분석 결과를 나타낸 것이다.6 shows the results of analysis of changes in the content of bone indicator factors in blood of Experimental Example 5.
도 7은 실험예 6에 따른 골 밀도 및 골 중 미네랄 농도 측정 결과를 나타낸 것이다.7 shows the results of measuring bone density and mineral concentration in bone according to Experimental Example 6.
도 8 실험예 7에 따른 경골에서 성장판 두께 측정 결과를 나타낸 것이다.Figure 8 shows the results of measuring the growth plate thickness in the tibia according to Experimental Example 7.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시하나, 하기 실시예는 본 발명을 예시하는 것일 뿐 본 발명의 범주 및 기술사상 범위 내에서 다양한 변경 및 수정이 가능함은 당업자에게 있어서 명백한 것이며, 이러한 변형 및 수정이 첨부된 특허청구범위에 속하는 것도 당연한 것이다.Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention, but the following examples are only illustrative of the present invention, and it is obvious to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It is natural that such modifications and modifications fall within the appended claims.
[실시예: 당근잎 추출물 제조][Example: Carrot Leaf Extract Preparation]
제조예 1: 당근잎 추출물의 제조Preparation Example 1: Preparation of carrot leaf extract
당근잎 500 g을 10배의 70% 에탄올로 3시간씩 2회에 걸쳐 환류 냉각 추출한 후 여과한 액을 감압농축기를 이용해 농축한 다음 동결건조하여 당근잎 추출물을 제조하였다.Carrot leaves 500 g were extracted with reflux cooling twice for 3 hours each with 10 times of 70% ethanol, and the filtered solution was concentrated using a vacuum concentrator and then freeze-dried to prepare a carrot leaf extract.
[실험예: 생체 외 실험][Experimental Example: In vitro experiment]
전조골세포주인 MC3T3-E1 세포는 5% FBS가 포함된 alpha-MEM 영양배지를 이용해 5% CO2, 37℃ 배양기에서 배양하였다. 골세포로의 분화는 10계대 이전의 세포에 100 nM 덱사메타손(Dexamethasone), 10 mM β-글리세로인산칼슘(β-glycerophosphate), 50 ㎍/㎖ 아스코르브산(ascorbic acid)이 포함된 분화배지(Conditioned Media, CM)로 교환하여 7, 14, 21일간 분화를 유도하였다.MC3T3-E1 cells, a precursor bone cell line, were cultured in a 5% CO 2 , 37°C incubator using an alpha-MEM nutrient medium containing 5% FBS. Differentiation into bone cells was conducted in a differentiation medium containing 100 nM dexamethasone, 10 mM β-glycerophosphate, and 50 μg/ml ascorbic acid in cells before passage 10. Media, CM) to induce differentiation for 7, 14 and 21 days.
또한, 골세포 분화에 있어 당근잎의 효과를 확인하기 위해 분화배지(DM)에 당근잎 추출물을 10, 50, 100 ㎍/㎖의 농도로 각각 처리하여 7, 14, 21일간 배양한 후, 인산분해효소 발현량, 칼슘염(calcium salt) 침착(Alizarin Red staining) 정도 및 분화 관련 유전자의 발현 측정을 통해 당근잎에 의한 골 분화 활성을 평가하였다.In addition, in order to check the effect of carrot leaves in osteocyte differentiation, the carrot leaf extract was treated in a differentiation medium (DM) at concentrations of 10, 50, and 100 ㎍/㎖, respectively, and cultured for 7, 14, and 21 days. Bone differentiation activity by carrot leaves was evaluated by measuring the expression level of degrading enzyme, the degree of calcium salt deposition (Alizarin Red staining), and expression of differentiation-related genes.
실험예 1: MC3T3-E1(preosteoblasts) 전조골세포에서 세포독성 효과 분석Experimental Example 1: Analysis of cytotoxic effects in MC3T3-E1 (preosteoblasts) proosteoblasts
당근잎 추출물의 세포 독성을 확인하기 위하여, 전조골세포 (MC3T3-E1)를 96 웰플레이트 (well-plate)에 1×104 cells/100㎕로 플래이팅(plating)하고, 다음날 당근잎 추출물을 농도별로 24시간 동안 배양하였다. 세포생존율을 측정하기 위하여 MTT 용액(최종농도 1 mg/㎖)을 가하고, 4시간 동안 배양한 후 DMSO (dimethyl sulfoxide)로 용해시켜 450 nm에서 흡광도를 마이크로 플레이트 리더(microplate reader)로 측정하여 그 결과를 도 1에 나타내었다. 이에 따르면, 당근잎 추출물을 MC3T3-E1 세포에 처리한 결과, 정상군(Con)과 비교하여 세포 독성에 영향을 미치지 않음을 확인하였다.In order to confirm the cytotoxicity of carrot leaf extract, plated precursor osteoblasts (MC3T3-E1) at 1×10 4 cells/100 μl in a 96 well plate, and the next day carrot leaf extract was Incubated for 24 hours by concentration. To measure the cell viability, MTT solution (final concentration 1 mg/ml) was added, incubated for 4 hours, dissolved in DMSO (dimethyl sulfoxide), and the absorbance at 450 nm was measured with a microplate reader. Is shown in Figure 1. According to this, as a result of treatment with the carrot leaf extract on MC3T3-E1 cells, it was confirmed that it did not affect cytotoxicity compared to the normal group (Con).
실험예 2: 염기성 인산분해효소(alkaline phosphatase, ALP) 발현량 측정 Experimental Example 2: Measurement of the expression level of alkaline phosphatase (ALP)
MC3T3-E1 세포를 12-웰 플레이트에 각 웰당 1x103 개씩 분주한 후 1일 후, 덱사메타손, 10 mM β-글리세로인산칼슘, 50 ㎍/㎖ 아스코르브산이 포함된 분화배지(CM)로 3, 5, 7일간 당근잎 추출물 100 ㎍/㎖와 함께 5% CO2, 37℃ 배양기에서 배양하였다. 일정 배양시간이 지난 후 배지를 수거하여 분석에 사용하였다. 분석은 Alkaline Phosphatase Assay Kit(Abcam, ab83369)를 사용하였으며 multiplate reader를 이용하여 405 nm에서 흡광도를 측정하였으며, 이에 따른 당근잎 추출물에 의한 ALP 발현량 측정 결과를 도 2에 나타내었다.One day after dispensing MC3T3-E1 cells into a 12-well plate, 1×10 3 cells per well, 3, 5 with differentiation medium (CM) containing dexamethasone, 10 mM β-glycerophosphate, and 50 μg/ml ascorbic acid , Carrot leaf extract 100 ㎍ / ㎖ for 7 days 5% CO 2 , was cultured in a 37 ℃ incubator. After a certain cultivation time, the medium was collected and used for analysis. For analysis, Alkaline Phosphatase Assay Kit (Abcam, ab83369) was used, and absorbance was measured at 405 nm using a multiplate reader, and the result of measuring the ALP expression level by the carrot leaf extract is shown in FIG. 2.
이에 따르면, 일반 배지(Normal media, NM)에 비하여 분화 배지(Conditioned media, CM)에서 ALP 활성이 증가함을 확인할 수 있었으며, 당근잎 추출물로 처리한 군(D)에서는 분화 배지보다 많은 양의 ALP 활성 증가를 확인하였다. 인산분해 효소 활성 결과를 통하여 당근잎 추출물이 골 분화 활성 효과가 있다는 가능성을 확보하였다. ALP는 세포의 파괴 정도를 나타내지만 세포 실험에서는 조골세포가 파괴되어야 새로운 세포가 생성되므로 ALP활성이 증가하는 것이 세포 성장이 높아지는 것이다.According to this, it was confirmed that ALP activity was increased in Conditioned media (CM) compared to normal media (NM), and in the group (D) treated with carrot leaf extract, a higher amount of ALP than the differentiation medium Increased activity was confirmed. Through the results of phosphatase activity, the possibility that carrot leaf extract has an effect on bone differentiation was secured. ALP shows the degree of destruction of cells, but in cell experiments, new cells are generated only when osteoblasts are destroyed, so that increasing ALP activity increases cell growth.
실험예 3: 칼슘 침착 평가Experimental Example 3: Calcium deposition evaluation
분화배지 및 분화배지와 당근잎을 농도별(10, 50, 100 ㎍/㎖)로 처리하여 배양된 세포를 인산염 식염수(PBS)로 세척하고 4% 포름알데히드로 10분간 고정하였다. 2% 알리자린 S(alizarin red S)(Millipore) 용액으로 5분간 염색한 후 탈염수로 여러 번 세척하여 여분의 염색액을 제거하고, 이에 따른 염색양상을 관찰하여 도 3에 나타내었다. 또한, 알리자린 S 를 정량화하기 위하여 100 mM 세틸피리듐 클로라이드(cetylpyridinium chloride) 용액을 첨가하여 상온에서 2시간 반응시켜 염색된 알리자린 S를 용해시켜 추출하였다. 용해된 알리자린 S의 농도는 multiplate reader를 이용하여 570 nm에서 흡광도를 측정하여 그 결과를 도 4에 나타내었다.Differentiation medium, differentiation medium, and carrot leaves were treated at different concentrations (10, 50, 100 ㎍/㎖), and the cultured cells were washed with phosphate saline (PBS) and fixed with 4% formaldehyde for 10 minutes. After staining with 2% alizarin red S (Millipore) solution for 5 minutes, washed several times with demineralized water to remove excess staining solution, and the resulting staining pattern was observed and shown in FIG. 3. In addition, in order to quantify Alizarin S, a 100 mM cetylpyridinium chloride solution was added and reacted at room temperature for 2 hours to dissolve and extract dyed Alizarin S. The concentration of dissolved Alizarin S was measured at 570 nm using a multiplate reader, and the results are shown in FIG. 4.
이에 따라 MC3T3-E1 세포에서 알리자린 레드 염색법 통해 당근잎에 의한 골세포 분화 활성 즉, 칼슘 침착을 비교한 결과를 살펴보면, Normal은 일반 영양배지로 배양한 음성 대조군이며, Control은 분화배지(DM)에 아무 것도 처리하지 않은 양성 대조군이고, CM(Conditioned media)+당근잎은 분화배지(DM)에 10, 50, 100 ㎍/㎖의 당근잎 추출물을 처리한 당근잎 처리군이다. Control군과 비교해 당근잎 처리군은 처리기간(7, 14, 21일)과 처리 농도 의존적으로 칼슘 침착에서 유의적인 증가가 관찰되었다. **, ***는 양성 대조군 대비 당근잎 처리군의 골세포 분화 활성이 통계적으로 유의미하게 증가하였다는 것으로, p<0.01, p<0.001 이고, ###는 분화배지 처리군 대비 처리기간에 따른 당근잎 처리군이 통계적으로 유의미하게 골세포 분화 활성이 증가하였다는 것으로, p<0.001이다.Accordingly, looking at the results of comparing the bone cell differentiation activity, that is, calcium deposition, by carrot leaves through the Alizarin Red staining method in MC3T3-E1 cells, Normal is a negative control cultured with normal nutrient medium, and Control is a differentiation medium (DM). It was a positive control without any treatment, and CM (Conditioned media) + carrot leaf was a carrot leaf-treated group treated with 10, 50, 100 ㎍/㎖ carrot leaf extract in differentiated medium (DM). Compared to the control group, the carrot leaf treatment group showed a significant increase in calcium deposition depending on the treatment period (7, 14, 21 days) and treatment concentration. **, *** indicates that the bone cell differentiation activity of the carrot leaf treatment group was statistically significantly increased compared to the positive control group, p<0.01, p<0.001, and ### during the treatment period compared to the differentiation medium treatment group. According to the results, the carrot leaf treatment group showed a statistically significant increase in osteoblast differentiation activity, p<0.001.
실험예 4: 골세포 분화 관련 유전자의 발현 평가Experimental Example 4: Evaluation of expression of genes related to bone cell differentiation
골성장과 관련된 특정 유전자(runx2, osteocalcin, colla1a)의 발현은 real-time PCR을 이용해 분석하였다. 6 웰 플레이트에 1 × 105개의 세포를 분주하여 7, 14, 21일 동안 배양하였으며 배양된 세포 에서 배지를 제거하고 RNA 추출키트(extraction kit) (Qiagen)를 사용하여 총 RNA를 추출하였다. cDNA 제작을 위해 cDNA 합성키트(synthesis kit)(Bio-Rad)를 이용하여 1 ㎍의 RNA 시료와 혼합한 후, thermal cycler(Bio-rad)를 이용해 cDNA를 합성하였다. 이렇게 얻은 cDNA 1 ㎕와 10 pmole의 프라이머를 sybr green master mix (Thermo Fisher)와 혼합하여 Thermal cycler(Thermo Fisher)를 이용해 real-time PCR을 수행하였다. 각각의 특정 유전자에 대한 프라이머의 염기서열은 다음과 같다. 실험군마다 3번 반복 수행하여 통계프로그램(Prism)을 이용해 도표화 하고 그 결과를 도 5에 나타내었다.The expression of specific genes (runx2, osteocalcin, colla1a) related to bone growth was analyzed using real-time PCR. 1 × 10 5 cells were dispensed into a 6-well plate and cultured for 7, 14, and 21 days. The medium was removed from the cultured cells, and total RNA was extracted using an RNA extraction kit (Qiagen). For cDNA production, a cDNA synthesis kit (Bio-Rad) was used to mix with 1 μg of RNA sample, and then cDNA was synthesized using a thermal cycler (Bio-rad). The obtained cDNA 1 µl and 10 pmole of primer were mixed with sybr green master mix (Thermo Fisher), and real-time PCR was performed using a thermal cycler (Thermo Fisher). The base sequence of the primers for each specific gene is as follows. It was repeated three times for each experimental group and plotted using a statistical program (Prism), and the results are shown in FIG. 5.
아래의 표 1에 상기 프라이머 정보를 정리하였다.The primer information is summarized in Table 1 below.
유전자gene 서열번호Sequence number forward sequence(5’→3’)forward sequence(5'→3') 서열번호Sequence number reward sequence(5’→3’)reward sequence(5'→3')
runx2runx2 1One GGT TGA CCT TTG TCC CAA TGGGT TGA CCT TTG TCC CAA TG 22 GTG TTT CGG TTT GCT ACT GGGTG TTT CGG TTT GCT ACT GG
osteocalcinosteocalcin 33 GCT TAA CCC TGC TTG TGAGCT TAA CCC TGC TTG TGA 44 TCC TAA ATA GTG ATA CCG TAG ATGTCC TAA ATA GTG ATA CCG TAG ATG
colla1colla1 55 GTG AGA CAG GCG AAC AAGGTG AGA CAG GCG AAC AAG 66 CAG GAG AAC CAG GAG GACCAG GAG AAC CAG GAG GAC
gapdhgapdh 77 TCT CCT GCG ACT TCA ACATCT CCT GCG ACT TCA ACA 88 CTG TAG CCG TAT TCA TTG TCCTG TAG CCG TAT TCA TTG TC
이에 따르면, MC3T3-E1 세포에서 real-time PCR을 통해 당근잎 추출물에 의한 골세포 분화 관련 유전자의 발현을 비교한 결과이다. Control은 분화배지(DM)에 아무것도 처리하지 않은 대조군이고, 10, 50, 100 ㎍/㎖은 분화배지와 각각의 농도별 당근잎 처리군이다. 대조군과 비교해 당근잎 처리군은 처리기간(7, 14, 21일)과 처리 농도 의존적으로 골세포 분화 유전자인 runx2, osteocalcin, colla1의 mRNA의 발현이 유의적으로 증가되었다. *, **, ***는 대조군 대비 당근잎 처리군의 골세포 분화 관련 유전자의 활성이 통계적으로 유의미하게 증가하였다는 것으로, p<0.05, p<0.01, p<0.001 이고, #, ##는 대조군 대비 당근잎 처리기간에 따라 통계적으로 유의미하게 골세포 분화 관련 유전자의 발현이 증가하였다는 것으로, p<0.05, p<0.01 이다.According to this, it is the result of comparing the expression of genes related to bone cell differentiation by carrot leaf extract through real-time PCR in MC3T3-E1 cells. Control is a control group that has not been treated with any differentiation medium (DM), and 10, 50, and 100 ㎍/㎖ are differentiation medium and carrot leaf treatment group for each concentration. Compared to the control group , the expression of mRNA of the bone cell differentiation genes runx2, osteocalcin, and colla1 was significantly increased in the treatment period (7, 14, 21 days) and treatment concentration-dependently. *, **, *** indicates that the activity of genes related to bone cell differentiation in the carrot leaf treatment group was statistically significantly increased compared to the control group, p<0.05, p<0.01, p<0.001, #, ## Is that the expression of bone cell differentiation-related genes was statistically significantly increased according to the treatment period of carrot leaves compared to the control group, p<0.05, p<0.01.
[실시예: 당근잎 추출물 투여][Example: Carrot Leaf Extract Administration]
실시예 1Example 1
상기 제조예 1에 따라 제조한 당근잎 추출물의 골 밀도, 골중 미네랄 농도 및 성장판 두께의 개선 효능을 확인하기 위하여, 골 성장이 저해된 마우스 동물모델을 제작하였다. In order to confirm the efficacy of improving the bone density, mineral concentration in bone, and growth plate thickness of the carrot leaf extract prepared according to Preparation Example 1, a mouse animal model with inhibited bone growth was prepared.
자세하게는 온도 및 습도가 관리되는 SPF(specific pathogen free)등급의 사육실에서 4주령의 마우스를 입고 후 일주일간 순화를 실시하였다. 일주일간 순화된 5주령의 마우스를 비타민 D 및 칼슘이 결핍된 식이를 8주간 실시하여 골 성장의 감소를 유발하였으며, 상기 제조예 1에 따라 제조한 당근잎 추출물을 동 기간 경구 투여하여 해당 추출물의 골 성장 저해 개선에 대한 효능을 확인하였다. 그리고 주 2회 체중 및 식이의 양을 측정하여 해당 모델의 건강 상태와 비타민D 및 칼슘이 결핍된 사료를 섭취가 올바르게 진행되는지 확인하였다. In detail, a 4-week-old mouse was worn in a SPF (specific pathogen free) grade breeding room where temperature and humidity were controlled, and then acclimatized for a week. Five-week-old mice purified for one week were fed a diet deficient in vitamin D and calcium for 8 weeks to induce a decrease in bone growth, and the carrot leaf extract prepared according to Preparation Example 1 was administered orally for the same period of time. Efficacy for improving inhibition of bone growth was confirmed. In addition, by measuring the weight and the amount of diet twice a week, it was confirmed that the health status of the model and the intake of diets deficient in vitamin D and calcium were properly progressed.
[실험예: 생체 내 실험][Experimental Example: In vivo experiment]
실험예 5: 혈중 골 지표 인자 함량 변화 분석Experimental Example 5: Analysis of changes in the content of bone indicator factors in blood
혈중 오스테오칼신, 염기성 인산분해효소(ALP)의 함량 분석을 위하여, 모델 동물을 아르베틴(Avertin)으로 마취하여 복대동맥에서 전혈을 채취하였다. 채혈한 전혈은 SST 튜브에 옮긴 후 충분히 섬유소에 의한 혈액응고가 진행되면 4,000rpm에서 10분 동안 원심분리하고, 적혈구의 용혈 등에 주의하여 상층액인 혈청을 취하여 혈중 오스테오칼신, 염기성 인산분해효소의 함량 분석에 사용하였으며, 사용하고 남은 혈청의 경우 초저온 냉동고에 보관하였다. 혈중 오스테오칼신 및 염기성 인산분해효소의 함량 분석은 ELISA(enzyme-linked immuosorbent assay) 분석을 통해 실시하였고, 그 결과를 도 6에 나타내었다. 여기서, 실험군의 구성은 정상대조군(Vehicle, 무처치군), 양성대조군 (Positive, 골 성장 저해 유도 및 Somatotrophin 0.1 mg/kg 처치군), 당근잎 저농도 처리군(KIOM_D_L, 골 성장 저해 유도 및 당근잎 150 mg/kg 처치군), 당근잎 고농도 처리군(KIOM_D_H, 골 성장 저해 유도 및 당근잎 450 mg/kg 처치군) 이다.In order to analyze the content of osteocalcin and basic phosphatase (ALP) in blood, a model animal was anesthetized with avertin, and whole blood was collected from the abdominal aorta. After transferring the collected whole blood to an SST tube, centrifuge for 10 minutes at 4,000 rpm when blood coagulation by fibrin has progressed sufficiently, and take the serum as the supernatant, paying attention to hemolysis of red blood cells, and analyze the content of osteocalcin and basic phosphatase in the blood. And the remaining serum was stored in a cryogenic freezer. Analysis of the content of osteocalcin and basic phosphatase in blood was performed through an enzyme-linked immuosorbent assay (ELISA) analysis, and the results are shown in FIG. 6. Here, the composition of the experimental group is a normal control group (Vehicle, no treatment group), a positive control group (positive, bone growth inhibition induction and Somatotrophin 0.1 mg/kg treatment group), carrot leaf low concentration treatment group (KIOM_D_L, bone growth inhibition induction and carrot leaf 150 mg/kg treatment group), carrot leaf high concentration treatment group (KIOM_D_H, bone growth inhibition induction and carrot leaf 450 mg/kg treatment group).
이에 따르면, 당근잎에 의해 혈청 내 오스테오칼신의 양이 유의성있게 증가하였으며, 또한 염기성 인산분해효소(ALP)의 양이 유의성 있게 감소하였음을 확인하였다. 또한 오스테오칼신의 경우, 양성 대조군으로 사용된 Somatotrophin 과 비교하였을 때에도 유의성 있게 증가함을 확인하였다. According to this, it was confirmed that the amount of osteocalcin in the serum was significantly increased by carrot leaves, and the amount of basic phosphatase (ALP) was significantly decreased. In addition, it was confirmed that osteocalcin significantly increased when compared to Somatotrophin used as a positive control.
실험예 6: 골 밀도 및 골 중 미네랄 농도 측정Experimental Example 6: Measurement of bone density and mineral concentration in bone
실험 종료 후 발골된 대퇴골 및 경골을 이용하여 골 밀도, 골중 미네랄 농도 및 성장판 두께를 측정하였다. 구체적으로 해당 분석에서 사용된 대퇴골의 골 밀도 및 골중 미네랄 농도는 동물 부검 후 10% 중성 포르말린에 3일간 고정 후 뼈를 제외한 근육 및 혈관 조직들의 제거한 후 Dual-energy X-ray absorptiometry(DEXA; Norland pDEXA Sabre; WI, USA)를 이용하여 측정한 후 Sabre Research(v3.6) 소프트웨어를 이용하여 분석하였고, 당근잎 추출물에 의한 대퇴골의 골 밀도 및 골 중 미네랄 농도 측정 결과를 도 7에 나타내었다. 여기서, 실험군의 구성은 정상대조군(Vehicle, 무처치군), 음성대조군(Negative, 골 성장 저해 유도군), 양성대조군 (골 성장 저해 유도 및 Somatotrophin 0.1 mg/kg 처치군), 당근잎 저농도 처리군(KIOM_D_L, 골 성장 저해 유도 및 당근잎 150 mg/kg 처치군), 당근잎 고농도 처리군(KIOM_D_H, 골 성장 저해 유도 및 당근잎 450 mg/kg 처치군) 이다.After the end of the experiment, bone density, bone mineral concentration, and growth plate thickness were measured using the deformed femur and tibia. Specifically, the bone density and bone mineral concentration of the femur used in the analysis were fixed in 10% neutral formalin for 3 days after an animal autopsy, and then muscle and vascular tissues other than the bone were removed, followed by Dual-energy X-ray absorptiometry (DEXA; Norland pDEXA). Saber; WI, USA), and then analyzed using Saber Research (v3.6) software, and the results of measuring bone density and mineral concentration in the femur by carrot leaf extract are shown in FIG. 7. Here, the composition of the experimental group is a normal control group (Vehicle, no treatment group), a negative control group (Negative, bone growth inhibition induction group), a positive control (bone growth inhibition induction and Somatotrophin 0.1 mg/kg treatment group), carrot leaf low concentration treatment group (KIOM_D_L, bone growth inhibition induction and carrot leaf 150 mg/kg treatment group), carrot leaf high concentration treatment group (KIOM_D_H, bone growth inhibition induction and carrot leaf 450 mg/kg treatment group).
이에 따르면, 당근잎 추출물을 투여하지 않은 음성 대조군 마우스에 비하여, 당근잎 추출물을 투여한 마우스의 골 밀도 및 골중 미네랄 농도가 증가함을 확인하였다. 또한, 상기 당근잎 추출물의 골 밀도 및 골중 미네랄 농도 증가 효과에 대한 양성 대조군으로 사용된 Somatotrophin를 투여한 마우스와 유사한 효과를 보이는 것으로 나타났다.According to this, it was confirmed that the bone density and bone mineral concentration of the mice to which the carrot leaf extract was administered were increased compared to the negative control mice to which the carrot leaf extract was not administered. In addition, it was found that the carrot leaf extract showed an effect similar to that of mice administered Somatotrophin, which was used as a positive control for the effect of increasing bone density and mineral concentration in bone.
실험예 7: 경골 성장판 두께 측정Experimental Example 7: Measurement of Tibial Growth Plate Thickness
실험 종료 후 경골에서 성장판 두께는 상기 방법과 전 처리 후 탈회, 포매, 박절을 통한 5㎛ 파라핀 절편을 제작하여 헤마톡실린-에오신 염색(hematoxylin & eosin stain)을 이용하여 성장판 두께를 관찰하였으며, 분석 소프트웨어 cellSens standard (Olympus)를 이용하여 성장판 두께를 측정하여 그 결과를 도 8에 나타내었다. 여기서, 실험군의 구성은 정상대조군(Vehicle, 무처치군), 음성대조군(Negative, 골 성장 저해 유도군), 양성대조군 (골 성장 저해 유도 및 Somatotrophin 0.1 mg/kg 처치군), 당근잎 고농도 처리군(KIOM_D_H, 골 성장 저해 유도 및 당근잎 450 mg/kg 처치군) 이다.After the end of the experiment, the thickness of the growth plate in the tibia was determined by using the above method and pretreatment, followed by demineralization, embedding, and thinning to make 5 μm paraffin sections, and observe the growth plate thickness using hematoxylin & eosin stains. The thickness of the growth plate was measured using the software cellSens standard (Olympus), and the results are shown in FIG. 8. Here, the composition of the experimental group is a normal control group (Vehicle, no treatment group), a negative control group (Negative, bone growth inhibition induction group), a positive control (bone growth inhibition induction and Somatotrophin 0.1 mg/kg treatment group), carrot leaf high concentration treatment group (KIOM_D_H, bone growth inhibition induction and carrot leaf 450 mg/kg treatment group).
이에 따르면, 당근잎 추출물을 투여하지 않은 음성대조군 마우스에 비하여, 상기 추출물을 투여한 마우스의 성장판 두께가 더 두꺼운 것으로 관찰되었으며, 상기 당근잎 추출물의 성장판 두께 증가 효과에 대한 양성대조군으로 사용된 Somatotrophin를 투여한 마우스와 유사한 효과를 보이는 것으로 나타났다.Accordingly, it was observed that the growth plate thickness of the mice administered with the extract was thicker than that of the negative control mice not administered with the carrot leaf extract, and Somatotrophin used as a positive control for the effect of increasing the growth plate thickness of the carrot leaf extract was used. It was found to exhibit similar effects to the administered mice.
이상, 본 발명의 실시예들에 대하여 설명하였으나, 해당 기술 분야에서 통상의 지식을 가진 자라면 특허청구범위에 기재된 본 발명의 사상으로부터 벗어나지 않는 범위 내에서, 구성 요소의 부가, 변경, 삭제 또는 추가 등에 의해 본 발명을 다양하게 수정 및 변경시킬 수 있을 것이며, 이 또한 본 발명의 권리범위 내에 포함된다고 할 것이다.In the above, embodiments of the present invention have been described, but those of ordinary skill in the art will add, change, delete or add components within the scope not departing from the spirit of the present invention described in the claims. Various modifications and changes can be made to the present invention by means of the like, and this will also be said to be included within the scope of the present invention.
이하, 본 발명을 상세하게 설명한다. Hereinafter, the present invention will be described in detail.
본 발명의 뼈 성장 촉진용 조성물은 당근잎 추출물을 유효성분으로 함유한다.The composition for promoting bone growth of the present invention contains carrot leaf extract as an active ingredient.
상기 당근잎은 추출용매와 1 : 5 내지 25, 바람직하게는 1 : 8 내지 15의 중량비로 혼합하여 40 내지 70 ℃, 바람직하게는 50 내지 60 ℃에서 10 내지 20시간, 바람직하게는 15 내지 18시간 동안 추출한 후 감압농축을 수행하여 추출물을 제조한다. 상기 당근잎과 추출용매의 중량비가 상기 범위를 벗어나는 경우에는 추출물에 당근잎의 유효성분이 적은 양으로 추출될 수 있다.The carrot leaves are mixed with an extraction solvent in a weight ratio of 1: 5 to 25, preferably 1: 8 to 15, at 40 to 70°C, preferably 50 to 60°C for 10 to 20 hours, preferably 15 to 18 After extraction for a period of time, concentration under reduced pressure is performed to prepare an extract. When the weight ratio of the carrot leaf and the extraction solvent is out of the above range, the active ingredient of carrot leaf may be extracted in a small amount in the extract.
상기 각 추출물을 추출하는 추출용매는 물, 탄소수 1 내지 4의 저급알코올, 에틸렌글리콜, 에틸에테르 또는 이들의 혼합용매이다. 상기 저급알코올로는 20 내지 80%의 메탄올, 에탄올, 부탄올 또는 프로판올을 들 수 있다.The extraction solvent for extracting each extract is water, a lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixed solvent thereof. The lower alcohol may include 20 to 80% of methanol, ethanol, butanol or propanol.
상기 추출용매로는 특별히 한정하는 것은 아니지만 60 내지 80%의 에탄올 수용액으로 추출된 추출물이 뼈 성장 촉진에 바람직하게 작용한다. The extraction solvent is not particularly limited, but the extract extracted with an aqueous solution of 60 to 80% ethanol preferably acts on promoting bone growth.
본 명세서에서 당근잎을 언급하면서 사용되는 용어 '추출물'은 추출용매를 처리하여 얻은 조추출물뿐만 아니라 당근잎 추출물의 가공물도 포함한다. 예를 들어, 당근잎 추출물은 감압 증류 및 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수 있다.The term'extract' used in referring to carrot leaves in the present specification includes not only crude extract obtained by treatment with an extraction solvent, but also processed products of carrot leaf extract. For example, the carrot leaf extract may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze drying or spray drying.
또한, 본 발명의 당근잎 추출물은 광의로는 당근잎을 동물에게 투여할 수 있도록 제형화된 당근잎 가공물, 예컨대, 당근잎 분말도 포함하는 의미를 갖는다. 비록 본 발명에서 당근잎으로 실험을 진행하긴 하였으나, 당근잎 가공물과 같은 형태로도 목적하는 효과를 달성할 수 있음은 당업자라면 예상 가능할 것이다.In addition, the carrot leaf extract of the present invention has a meaning to include a carrot leaf processed product, for example, carrot leaf powder, formulated so that carrot leaf can be administered to animals. Although the experiment was carried out with carrot leaves in the present invention, it will be expected by those skilled in the art that the desired effect can be achieved even in the same form as a processed carrot leaf product.
한편, 본 명세서에서 용어 '유효성분으로 포함하는'이란 당근잎 추출물의 효능 또는 활성을 달성하는 데 충분한 양을 포함하는 것을 의미한다. 일예로, 상기 당근잎 추출물은 10 내지 1500 ㎍/㎖, 바람직하게는 100 내지 1000 ㎍/㎖의 농도로 사용된다. 당근잎 추출물은 천연물로서 과량 투여하여도 인체에 부작용이 없으므로 본 발명의 조성물 내에 포함되는 당근잎 추출물의 양적 상한은 당업자가 적절한 범위 내에서 선택하여 실시할 수 있다.Meanwhile, in the present specification, the term "including as an active ingredient" means including an amount sufficient to achieve the efficacy or activity of the carrot leaf extract. For example, the carrot leaf extract is used in a concentration of 10 to 1500 µg/ml, preferably 100 to 1000 µg/ml. Since carrot leaf extract is a natural product and does not have side effects on the human body even if it is administered in excess, the upper limit of the quantity of the carrot leaf extract contained in the composition of the present invention can be selected and carried out by a person skilled in the art within an appropriate range.
본 발명은 당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 식품 조성물을 제공한다.The present invention provides a food composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
또한 당근잎 추출물은 뼈 성장을 촉진하므로 성장 장애의 예방 또는 개선 용도로 사용될 수 있다. 상기 성장 장애는 정상 변이 저신장증 또는 질병에 의해 2차적으로 오는 저신장증일 수 있다. 상기 정상 변이 저신장증은 가족성 저신장, 체질적 성장 지연, 협의의 특발성 저신장일 수 있다. 또한, 질병에 의해 2차적으로 오는 저신장증은 1차성 성장 장애(내인성 장애) 또는 2차성 성장 장애(외인성 성장 장애)일 수 있으며, 상기 1차성 성장 장애로는 골연골 이형성증, 염색체 이상(다운 증후군 또는 터너 증후군)에 의한 저신장, 부당 경량아(자궁 내 성장 지연), 프레더-윌리 증후군에 의한 저신장, 러셀-실버 증후군에 의한 저신장, 누난 증후군에 의한 저신장이 있으며, 상기 2차성 성장 장애로는 만성 전신성 질환에 의한 저신장, 성장호르몬 결핍증에 의한 저신장, 갑상샘 저하증에 의한 저신장, 쿠싱증후군에 의한 저신장, 정신사회적 왜소증이 있다.In addition, since carrot leaf extract promotes bone growth, it can be used for preventing or improving growth disorders. The growth disorder may be normal variant short stature or short stature secondary to disease. The normal variant short stature may be familial short stature, delayed constitutional growth, or negotiated idiopathic short stature. In addition, short kidney disease secondary to the disease may be a primary growth disorder (endogenous disorder) or a secondary growth disorder (exogenous growth disorder), and the primary growth disorder includes osteochondral dysplasia, chromosomal abnormalities (Down syndrome or Turner syndrome), unfair lightweight infants (delayed growth in the uterus), short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, and short stature due to Noonan syndrome, and the secondary growth disorders are chronic There are short stature due to systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, short stature due to Cushing's syndrome, and psychosocial dwarfism.
본 발명에 따른 식품 조성물은 하기 약학 조성물과 동일한 방식으로 제제화되어 기능성 식품으로 이용하거나, 각종 식품에 첨가할 수 있다. 본 발명의 조성물을 첨가할 수 있는 식품으로는 예를 들어, 음료류, 알코올 음료류, 과자류, 다이어트바, 유제품, 육류, 초코렛, 피자, 라면, 기타 면류, 껌류, 아이스크림류, 비타민 복합제, 건강보조식품류 등이 있다.The food composition according to the present invention may be formulated in the same manner as the following pharmaceutical composition to be used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, health supplements. Etc.
본 발명의 식품 조성물은 유효성분으로서 당근잎 추출물뿐만 아니라, 식품 제조 시에 통상적으로 첨가되는 성분을 포함할 수 있으며, 예를 들어, 단백질, 탄수화물, 지방, 영양소, 조미제 및 향미제를 포함한다. 상술한 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스, 올리고당 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 향미제로서 천연 향미제 [타우마틴, 스테비아 추출물 (예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스파르탐 등)를 사용할 수 있다. 예컨대, 본 발명의 식품 조성물이 드링크제와 음료류로 제조되는 경우에는 본 발명의 당근잎 추출물 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙, 및 각종 식물 추출액 등을 추가로 포함시킬 수 있다.The food composition of the present invention may include not only carrot leaf extract as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, flavoring agents and flavoring agents. . Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, in addition to the carrot leaf extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included. .
본 발명은 상기 당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 식품 조성물을 포함하는 건강기능식품을 제공한다. 건강기능식품이란, 당근잎 추출물을 음료, 차류, 향신료, 껌, 과자류 등의 식품소재에 첨가하거나, 캡슐화, 분말화, 현탁액 등으로 제조한 식품으로, 이를 섭취할 경우 건강상 특정한 효과를 가져오는 것을 의미하나, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용시 발생할 수 있는 부작용 등이 없는 장점이 있다. 이와 같이 하여 얻어지는 본 발명의 건강기능식품은, 일상적으로 섭취하는 것이 가능하기 때문에 매우 유용하다. 이와 같은 건강기능식품에 있어서의 당근잎 추출물의 첨가량은, 대상인 건강기능식품의 종류에 따라 달라 일률적으로 규정할 수 없지만, 식품 본래의 맛을 손상시키지 않는 범위에서 첨가하면 되며, 대상 식품에 대하여 통상 0.01 내지 50 중량%, 바람직하기로는 0.1 내지 20 중량%의 범위이다. 또한, 환제, 과립제, 정제 또는 캡슐제 형태의 건강기능식품의 경우에는 통상 0.1 내지 100 중량% 바람직하기로는 0.5 내지 80 중량%의 범위에서 첨가하면 된다. 한 구체예에서, 본 발명의 건강기능식품은 환제, 정제, 캡슐제 또는 음료의 형태일 수 있다.The present invention provides a health functional food comprising a food composition for promoting bone growth comprising the carrot leaf extract as an active ingredient. Health functional foods are foods prepared by adding carrot leaf extract to food ingredients such as beverages, teas, spices, chewing gums, confectionery, or encapsulating, powdering, and suspending. However, unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time by using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of carrot leaf extract added in such health functional foods cannot be uniformly regulated depending on the type of health functional food to be targeted, but can be added within the range that does not damage the original taste of the food. It is in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.
또한, 본 발명은 당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 약학 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
또한, 당근잎 추출물은 뼈 성장을 촉진하므로 성장 장애의 예방 또는 치료 용도로 사용될 수 있다. 상기 성장 장애는 정상 변이 저신장증 또는 질병에 의해 2차적으로 오는 저신장증일 수 있다. 상기 정상 변이 저신장증은 가족성 저신장, 체질적 성장 지연, 협의의 특발성 저신장일 수 있다. 또한, 질병에 의해 2차적으로 오는 저신장증은 1차성 성장 장애(내인성 장애) 또는 2차성 성장 장애(외인성 성장 장애)일 수 있으며, 상기 1차성 성장 장애로는 골연골 이형성증, 염색체 이상(다운 증후군 또는 터너 증후군)에 의한 저신장, 부당 경량아(자궁 내 성장 지연), 프레더-윌리 증후군에 의한 저신장, 러셀-실버 증후군에 의한 저신장, 누난 증후군에 의한 저신장이 있으며, 상기 2차성 성장 장애로는 만성 전신성 질환에 의한 저신장, 성장호르몬 결핍증에 의한 저신장, 갑상샘 저하증에 의한 저신장, 쿠싱증후군에 의한 저신장, 정신사회적 왜소증이 있다.In addition, since the carrot leaf extract promotes bone growth, it can be used for preventing or treating growth disorders. The growth disorder may be normal variant short stature or short stature secondary to disease. The normal variant short stature may be familial short stature, delayed constitutional growth, or negotiated idiopathic short stature. In addition, short kidney disease secondary to the disease may be a primary growth disorder (endogenous disorder) or a secondary growth disorder (exogenous growth disorder), and the primary growth disorder includes osteochondral dysplasia, chromosomal abnormalities (Down syndrome or Turner syndrome), unfair lightweight infants (delayed growth in the uterus), short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, and short stature due to Noonan syndrome, and the secondary growth disorders are chronic There are short stature due to systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, short stature due to Cushing's syndrome, and psychosocial dwarfism.
본 발명의 약학 조성물은 상기 유효 성분 이외에 약제학적으로 적합하고 생리학적으로 허용되는 보조제를 사용하여 제조될 수 있으며, 상기 보조제로는 부형제, 붕해제, 감미제, 결합제, 피복제, 팽창제, 윤활제, 활택제 또는 향미제 등을 사용할 수 있다.The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, lubricants. Agents or flavoring agents can be used.
상기 약학 조성물은 투여를 위해서 상기 기재한 유효 성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 약학 조성물로 바람직하게 제제화할 수 있다.For administration, the pharmaceutical composition may include one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.
상기 약학 조성물의 제제 형태는 과립제, 산제, 정제, 피복정, 캡슐제, 좌제, 액제, 시럽, 즙, 현탁제, 유제, 점적제 또는 주사 가능한 액제 등이 될 수 있다. 예를 들어, 정제 또는 캡슐제의 형태로의 제제화를 위해, 유효 성분은 에탄올, 글리세롤, 물 등과 같은 경구, 무독성의 약제학적으로 허용 가능한 불활성 담체와 결합될 수 있다. 또한, 원하거나 필요한 경우, 적합한 결합제, 윤활제, 붕해제 및 발색제 또한 혼합물로 포함될 수 있다. 적합한 결합제는 이에 제한되는 것은 아니나, 녹말, 젤라틴, 글루코스 또는 베타-락토오스와 같은 천연 당, 옥수수 감미제, 아카시아, 트래커캔스 또는 소듐올레이트와 같은 천연 및 합성 검, 소듐 스테아레이트, 마그네슘 스테아레이트, 소듐 벤조에이트, 소듐 아세테이트, 소듐 클로라이드 등을 포함한다. 붕해제는 이에 제한되는 것은 아니나, 녹말, 메틸 셀룰로스, 아가, 벤토니트, 잔탄 검 등을 포함한다.The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.
액상 용액으로 제제화되는 조성물에 있어서 허용 가능한 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다.As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, sterilization and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.
본 발명의 약학 조성물은 경구 또는 비경구로 투여할 수 있고, 비경구 투여인 경우에는 정맥내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여 등으로 투여할 수 있으며, 바람직하게는 경구 투여이다.The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, and the like, preferably oral administration.
본 발명의 약학 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하며, 보통으로 숙련된 의사는 소망하는 치료 또는 예방에 효과적인 투여량을 용이하게 결정 및 처방할 수 있다. 본 발명의 바람직한 구현예에 따르면, 본 발명의 약학 조성물의 1일 투여량은 0.001-10 g/㎏이다.A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, and is usually As a result, a skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.
본 발명의 약학 조성물은 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액 또는 유화액 형태이거나 엑스제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention may be prepared in a unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient, or may be prepared by placing it in a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.
또한, 본 발명은 뼈 성장 촉진용 의약 또는 식품의 제조를 위한 당근잎 추출물의 용도를 제공한다. 상기한 바와 같이 당근잎 추출물은 뼈 성장 촉진과 뼈 성장이 저하와 관련된 질환의 치료 또는 개선을 위한 용도로 이용될 수 있다.In addition, the present invention provides a use of a carrot leaf extract for the manufacture of a medicine or food for promoting bone growth. As described above, the carrot leaf extract can be used for the treatment or improvement of diseases associated with promoting bone growth and reducing bone growth.
또한, 본 발명은 포유동물에게 유효량의 당근잎 추출물을 투여하는 것을 포함하는 장애 또는 불면증의 개선, 수면예방 또는 치료 방법을 제공한다.In addition, the present invention provides a method for improving, preventing or treating sleep, a disorder or insomnia comprising administering an effective amount of carrot leaf extract to a mammal.
여기에서 사용된 용어 "포유동물"은 치료, 관찰 또는 실험의 대상인 포유동물을 말하며, 바람직하게는 인간을 말한다.The term "mammal" as used herein refers to a mammal that is the subject of treatment, observation or experimentation, and preferably refers to a human.
여기에서 사용된 용어 "유효량"은 연구자, 수의사, 의사 또는 기타 임상의에 의해 생각되는 조직계, 동물 또는 인간에서 생물학적 또는 의학적 반응을 유도하는 유효 성분 또는 약학 조성물의 양을 의미하는 것으로, 이는 해당 질환 또는 장애의 증상의 완화를 유도하는 양을 포함한다. 본 발명의 유효 성분에 대한 유효량 및 투여횟수는 원하는 효과에 따라 변화될 수 있다. 그러므로, 투여될 최적의 투여량은 당업자에 의해 쉽게 결정될 수 있으며, 질환의 종류, 질환의 중증도, 조성물에 함유된 유효성분 및 다른 성분의 함량, 제형의 종류, 및 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 분비율, 치료기간, 동시 사용되는 약물을 비롯한 다양한 인자에 따라 조절될 수 있다. 본 발명의 예방, 치료 또는 개선 방법에 있어서, 성인의 경우, 추출물을 1일 1회 내지 수회 투여시, 0.001 g/kg 내지 10 g/kg의 용량으로 투여하는 것이 바람직하다.The term "effective amount" as used herein refers to an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, which is considered by a researcher, veterinarian, doctor or other clinician, which Or an amount that induces relief of symptoms of the disorder. The effective amount and the number of administrations for the active ingredient of the present invention may vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the amount of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient. It can be adjusted according to various factors including condition, sex and diet, time of administration, route of administration and secretion rate of the composition, duration of treatment, and drugs used simultaneously. In the prevention, treatment or improvement method of the present invention, in the case of an adult, it is preferable to administer the extract at a dose of 0.001 g/kg to 10 g/kg when administered once to several times a day.
본 발명의 치료방법에서 당근잎 추출물을 유효 성분으로 포함하는 조성물은 경구, 직장, 정맥내, 동맥내, 복강내, 근육내, 흉골내, 경피, 국소, 또는 피내 경로를 통해 통상적인 방식으로 투여할 수 있다.In the treatment method of the present invention, the composition containing the carrot leaf extract as an active ingredient is administered in a conventional manner through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, or intradermal routes. can do.
본 발명의 당근잎 추출물을 포함하는 뼈 성장 촉진용 조성물은 성장기에 있는 어린이나 청소년에 투여 또는 섭취시킬 경우 성장판 확장을 통해 뼈 성장을 향상시키고, 골밀도가 낮은 성인에 투여 또는 섭취시킬 경우에는 골밀도를 높이고 골 중 미네랄 농도를 향상시킬 수 있다.The composition for promoting bone growth comprising the carrot leaf extract of the present invention improves bone growth through growth plate expansion when administered or ingested to children or adolescents in the growing period, and increases bone density when administered or ingested to adults with low bone density. It can increase and improve the mineral concentration in the bone.

Claims (14)

  1. 당근잎 추출물 및 약제학적으로 허용 가능한 담체를 포함하는 뼈 성장 촉진용 식품 조성물.A food composition for promoting bone growth comprising a carrot leaf extract and a pharmaceutically acceptable carrier.
  2. 제1항에 있어서,The method of claim 1,
    상기 당근잎 추출물은 물, 탄소수 1-4의 알코올, 또는 이들의 혼합용매에 의해 추출된 것을 특징으로 하는 뼈 성장 촉진용 식품 조성물.The carrot leaf extract is a food composition for promoting bone growth, characterized in that extracted with water, alcohol having 1-4 carbon atoms, or a mixed solvent thereof.
  3. 제1항에 있어서,The method of claim 1,
    상기 뼈 성장 촉진용 식품 조성물은 runx2, osteocalcin, 및 colla1 중에서 선택된 1종 이상의 골세포 분화 유전자의 활성을 증가시키는 것을 특징으로 하는 뼈 성장 촉진용 식품 조성물.The food composition for promoting bone growth is a food composition for promoting bone growth, characterized in that to increase the activity of one or more bone cell differentiation genes selected from runx2, osteocalcin, and colla1.
  4. 제1항에 있어서,The method of claim 1,
    상기 뼈 성장 촉진용 식품 조성물은 가족성 저신장, 체질적 성장 지연, 골연골 이형성, 염색체 이상에 의한 저신장, 프레더-윌리 증후군에 의한 저신장, 러셀-실버 증후군에 의한 저신장, 누난 증후군에 의한 저신장, 만성 전신성 질환에 의한 저신장, 성장호르몬 결핍증에 의한 저신장, 갑상샘 저하증에 의한 저신장, 및 쿠싱증후군에 의한 저신장 중에서 선택된 어느 하나의 증상 개선을 위한 것인 뼈 성장 촉진용 식품 조성물.The food composition for promoting bone growth includes familial short stature, constitutional growth delay, osteocartilage dysplasia, short stature due to chromosomal abnormalities, short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, short stature due to Noonan syndrome, A food composition for promoting bone growth for improving any one symptom selected from short stature due to chronic systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, and short stature due to Cushing's syndrome.
  5. 제1항에 있어서,The method of claim 1,
    상기 뼈 성장 촉진용 식품 조성물은 과립제, 산제, 정제, 피복정, 캡슐제, 액제, 시럽, 즙, 현탁제, 유제 및 점적제 중에서 선택된 1종 이상으로 제제화되어 식품에 첨가되는 것을 특징으로 하는 뼈 성장 촉진용 식품 조성물.The bone growth promoting food composition is formulated as one or more selected from granules, powders, tablets, coated tablets, capsules, liquids, syrups, juices, suspensions, emulsions and drops, and added to food. Food composition for promoting growth.
  6. 당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 건강기능식품 조성물.A health functional food composition for promoting bone growth comprising carrot leaf extract as an active ingredient.
  7. 당근잎 추출물을 유효성분으로 포함하는 뼈 성장 촉진용 약학 조성물.A pharmaceutical composition for promoting bone growth comprising a carrot leaf extract as an active ingredient.
  8. 제7항에 있어서,The method of claim 7,
    상기 당근잎 추출물은 물, 탄소수 1-4의 알코올, 또는 이들의 혼합용매에 의해 추출된 것을 특징으로 하는 뼈 성장 촉진용 약학 조성물.The carrot leaf extract is a pharmaceutical composition for promoting bone growth, characterized in that extracted with water, alcohol having 1-4 carbon atoms, or a mixed solvent thereof.
  9. 제7항에 있어서,The method of claim 7,
    상기 뼈 성장 촉진용 약학 조성물은 runx2, osteocalcin, 및 colla1 중에서 선택된 1종 이상의 골세포 분화 유전자의 활성을 증가시키는 것을 특징으로 하는 뼈 성장 촉진용 약학 조성물.The pharmaceutical composition for promoting bone growth is a pharmaceutical composition for promoting bone growth, characterized in that to increase the activity of one or more bone cell differentiation genes selected from runx2, osteocalcin, and colla1.
  10. 제7항에 있어서,The method of claim 7,
    상기 뼈 성장 촉진용 약학 조성물은 가족성 저신장, 체질적 성장 지연, 골연골 이형성, 염색체 이상에 의한 저신장, 프레더-윌리 증후군에 의한 저신장, 러셀-실버 증후군에 의한 저신장, 누난 증후군에 의한 저신장, 만성 전신성 질환에 의한 저신장, 성장호르몬 결핍증에 의한 저신장, 갑상샘 저하증에 의한 저신장, 및 쿠싱증후군에 의한 저신장 중에서 선택된 어느 하나의 증상 치료를 위한 것인 뼈 성장 촉진용 약학 조성물.The pharmaceutical composition for promoting bone growth includes familial short stature, constitutional growth delay, osteocartilage dysplasia, short stature due to chromosomal abnormalities, short stature due to Freder-Willi syndrome, short stature due to Russell-Silver syndrome, short stature due to Noonan syndrome, A pharmaceutical composition for promoting bone growth for the treatment of any one symptom selected from short stature due to chronic systemic disease, short stature due to growth hormone deficiency, short stature due to hypothyroidism, and short stature due to Cushing's syndrome.
  11. 제1항에 있어서,The method of claim 1,
    상기 뼈 성장 촉진용 식품 조성물은 과립제, 산제, 정제, 피복정, 캡슐제, 액제, 시럽, 즙, 현탁제, 유제 및 점적제 중에서 선택된 1종 이상으로 제제화되는 것을 특징으로 하는 뼈 성장 촉진용 약학 조성물.The food composition for promoting bone growth is a pharmaceutical for promoting bone growth, characterized in that it is formulated as one or more selected from granules, powders, tablets, coated tablets, capsules, solutions, syrups, juices, suspensions, emulsions and drops. Composition.
  12. 뼈 성장 촉진용 의약 제조를 위한 당근잎 추출물의 용도. Use of carrot leaf extract for the manufacture of a medicine for promoting bone growth.
  13. 제12항에 있어서,The method of claim 12,
    상기 뼈 성장 촉진용 의약은 성장 장애의 예방 또는 치료용 의약인 것을 특징으로 하는 당근잎 추출물의 용도.The use of carrot leaf extract, characterized in that the medicament for promoting bone growth is a medicament for preventing or treating growth disorders.
  14. 성장 장애 환자에게 당근잎 추출물을 유효성분으로 포함하는 약학 조성물을 투여하는 것을 포함하는 성장 장애의 치료방법.A method for treating a growth disorder comprising administering a pharmaceutical composition comprising a carrot leaf extract as an active ingredient to a patient with a growth disorder.
PCT/KR2019/016384 2019-06-05 2019-11-26 Composition for promoting bone growth including carrot leaf as active ingredient WO2020246671A1 (en)

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KR100242730B1 (en) * 1997-11-03 2000-03-02 최재영 A sauce made of carrot leaves
KR20030011464A (en) * 2001-08-03 2003-02-11 성덕모 Composition for healthy food carpable of removing toxic oxygen using wild plants and method for extracting wild plants
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