WO2019009606A1 - Pharmaceutical composition containing mir-4507 microrna as active ingredient for prevention or treatment of dermatitis - Google Patents

Pharmaceutical composition containing mir-4507 microrna as active ingredient for prevention or treatment of dermatitis Download PDF

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WO2019009606A1
WO2019009606A1 PCT/KR2018/007562 KR2018007562W WO2019009606A1 WO 2019009606 A1 WO2019009606 A1 WO 2019009606A1 KR 2018007562 W KR2018007562 W KR 2018007562W WO 2019009606 A1 WO2019009606 A1 WO 2019009606A1
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dermatitis
microrna
composition
expression
mir
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PCT/KR2018/007562
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French (fr)
Korean (ko)
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유영
윤원석
강대중
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고려대학교 산학협력단
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy

Definitions

  • the present invention relates to a pharmaceutical composition for preventing or treating dermatitis containing miR-4507 microRNA as an active ingredient.
  • Atopic dermatitis is a chronic skin disease that is medically undetermined and frequently occurring in modern humans.
  • Atopic dermatitis is generally known as a disease caused by an imbalance of the human immune system.
  • Our body's immune system consists of humoral immunity mediated by B lymphocytes and cellular immunity mediated by T lymphocytes.
  • T-cells and Th-cells are involved in T-cells that mediate cell-mediated immunity, and Th-cells are divided into Th-1 and Th-2.
  • Th-2 When the amount of Th-2 is suddenly increased and the amount of Th-1 is decreased, the allergic diseases such as atopic dermatitis, allergic rhinitis and allergic asthma occur, Conversely, when the amount of Th-1 is increased, an autoimmune disease is caused.
  • the hypersensitivity of the Th-2 immune response is a key component of atopic skin disease.
  • the IL-4 and IL-13 stimulate the isotype of B cells and the production of IgE by the IL-4, IL-5 and IL-13 cytokines .
  • atopic dermatitis has developed extensively from childhood to adulthood, so it is urgent to develop a therapeutic agent.
  • steroid-based atopic dermatitis inhibitors are used as therapeutic agents.
  • they are not a primary solution for atopic dermatitis but show a temporary relief effect.
  • side effects that cause resistance are known.
  • CCL22 is a ligand of CCR4 that selectively expresses CD2 + T cells of Th-2 phenotype in T cells.
  • the concentration of CCL22 in the atopic dermatitis patients is rapidly increased compared with that of normal patients.
  • MDC is a chemokine that has been reported to significantly increase its serum concentration in patients with atopic dermatitis (Hijnen et al., J. Allergy Clin. Immunol. 113 (2) 334-340), used as a treatment for atopic dermatitis It has been reported that serum concentrations of MDC are decreased when ciclosporin A or corticosteroids are administered to patients with atopic dermatitis (Y. Shimada et al., J. Dermatol.
  • MDC is expressed in a large amount and a substance capable of inhibiting such expression can be used as a therapeutic agent for atopic dermatitis (Horikawa et. al., Int. Immunol., 14 (7) 767-773, 2002).
  • miRNA is a kind of small RNA (endogenous small RNA) of 20-25 nucleotides in a cell, which is derived from a DNA that does not synthesize proteins and is generated from a hairpin-shaped transcript .
  • miRNAs generally bind to the complementary sequence of the 3'-UTR of the target mRNA and induce the translation inhibition or destabilization of the mRNA, ultimately acting as a repressor to inhibit the protein synthesis of the target mRNA.
  • One miRNA targets several mRNAs, and mRNA is also known to be regulated by several miRNAs.
  • miRNAs have been of great interest in life sciences as they are known to play a very important role in many biological processes including developmental timing, apoptosis, lipid metabolism, and regulation of hematopoietic differentiation.
  • studies on the function of miRNAs in diseases such as cancer and development have progressed considerably, while studies on the role of miRNAs in dermatology have been relatively rare.
  • the inventors of the present invention conducted extensive studies to overcome the problems of the prior art. As a result, when the microRNA, which is a single base strand composed of about 21 to 25 nucleotides, was subcutaneously injected into the dermatitis-induced experimental group, Th-2 immune response In addition, the expression of IFN-y, an indicator of Th-1 immune response, is suppressed, and the production of MDC, an indicator of atopic disease, is suppressed and the immune response of IgE is rapidly reduced The present invention has been completed.
  • the main object of the present invention is to provide a pharmaceutical composition for preventing or treating dermatitis containing miR-4507 microRNA as an active ingredient.
  • a pharmaceutical composition for preventing or treating dermatitis which comprises miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
  • a pharmaceutical composition for preventing or treating dermatitis which comprises hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2 as an active ingredient.
  • the composition can inhibit the production of macrophage-derived chemokines (MDCs).
  • MDCs macrophage-derived chemokines
  • the composition can inhibit the expression of IgE.
  • the composition can inhibit the expression of IL-4.
  • the composition can induce the expression of IFN- ?.
  • a cosmetic composition for improving dermatitis which comprises miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
  • a cosmetic composition for improving dermatitis which comprises, as an active ingredient, hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2.
  • the composition can inhibit the expression of IgE.
  • the composition can inhibit the expression of IL-4.
  • the composition can induce the expression of IFN- ?.
  • an external preparation for skin for improving dermatitis comprising the composition according to an embodiment of the present invention.
  • the external preparation for skin may have a formulation selected from the group consisting of a gel, a cream, a milky lotion, a lotion, a pack, a lotion, an essence, a powder, a soap, a bath agent and a spray.
  • the pharmaceutical composition for the prevention or treatment of dermatitis of the present invention and the cosmetic composition for improving dermatitis reduce the expression of IL-4, which is an index of Th-2 immune response, and the expression of IFN-y, , Inhibiting the production of MDC, an indicator of atopic disease, and rapidly reducing the immune response of IgE. In addition, itching can be relieved, which is one of the typical symptoms of dermatitis.
  • miR-4507 microRNA is a human-derived RNA, it is a low-risk and safe substance for side effects, and thus the composition of the present invention containing it as an active ingredient can be usefully used as an external preparation for skin or cosmetics.
  • FIG. 1 is a view showing that a composition containing miR-4507 microRNA has an inflammation-reducing effect on the skin in a dermatitis-induced mouse.
  • FIG. 2 is a graph showing a result of inhibiting MDC, a chemokine that induces Th2 cells in a dermatitis-induced mouse, wherein the composition containing miR-4507 microRNA is inhibited.
  • Fig. 3 shows the effect of a composition containing miR-4507 microRNA to suppress itching in skin inflammation mice.
  • FIG. 4 is a graph showing the results of inhibiting the expression of IgE, a dermatitis-inducing antibody, in a composition containing miR-4507 microRNA.
  • FIG. 5 is a graph showing the results of inhibiting the expression of IL-4, a dermatitis-inducing cytokine, in a composition containing miR-4507 microRNA.
  • FIG. 6 is a graph showing a result of induction of a related IFN-gamma expression of dermatitis by a composition containing miR-4507 microRNA.
  • a pharmaceutical composition for preventing or treating dermatitis which comprises miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
  • a pharmaceutical composition for preventing or treating dermatitis which comprises hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2 as an active ingredient.
  • microRNA contained in the composition of the present invention is a nucleotide sequence of miR-4507 microRNA of 21 nucleotides, preferably microRNA of SEQ ID NO: 1 or SEQ ID NO: 2 shown in Table 1 below.
  • composition inhibits the formation of MDC (macrophage-derived chemokine), inhibits the expression of IgE and IL-4, and can induce the expression of IFN- ?.
  • MDC macrophage-derived chemokine
  • a microRNA polynucleotide sequence according to the present invention may be operably linked to an expression control sequence, wherein the operably linked gene sequence and expression control sequence are an expression vector containing a selectable marker and a replication origin Lt; / RTI >
  • Operably linked may be a gene and expression control sequence linked in such a way as to enable gene expression when the appropriate molecule is coupled to the expression control sequence.
  • Expression control sequence means a DNA sequence that regulates the expression of a polynucleotide sequence operably linked to a particular host cell.
  • regulatory sequences include promoters for conducting transcription, any operator sequences for regulating transcription, sequences encoding suitable mRNA ribosome binding sites, sequences controlling the termination of transcription and translation.
  • MiR-4507 microRNA which induces the dermatitis prevention and therapeutic effect of the present invention, can be produced by commonly used gene synthesis, and when the produced microRNA is introduced into the body, the miRNA is also expressed by a viral vector or a plasmid vector .
  • the pharmaceutical composition of the present invention may be prepared by a method known in the pharmaceutical art to be used as a pharmaceutical agent and may be prepared by mixing with a pharmaceutically acceptable carrier, an excipient, a diluent, a stabilizer, Capsules, injections, and the like.
  • the diluent may be a non-aqueous solvent such as propylene glycol, polyethylene glycol, vegetable oil such as olive oil or peanut oil, or water (preferably 0.05 M phosphate buffer) containing saline (preferably 0.8% saline) , And the like, but the present invention is not limited thereto.
  • a non-aqueous solvent such as propylene glycol, polyethylene glycol, vegetable oil such as olive oil or peanut oil, or water (preferably 0.05 M phosphate buffer) containing saline (preferably 0.8% saline) , And the like, but the present invention is not limited thereto.
  • excipient examples include starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, anhydrous skimmed milk, glycerol, , Ethanol, and the like, but are not limited thereto.
  • stabilizer examples include proteins such as carbohydrates such as sorbitol, mannitol, starch, sucrose, dextran, glutamate and glucose and animal, vegetable or microbial proteins such as milk powder, serum albumin and casein. no.
  • preservative examples include, but are not limited to, thimerosal, mertiolate, gentamicin, neomycin, nystatin, amphotericin B, tetracycline, penicillin, streptomycin, polyamicin B and the like.
  • the pharmaceutical composition may be administered as an individual therapeutic agent or in combination with another therapeutic agent, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by a person skilled in the art.
  • parenterally e. G., Intravenously, subcutaneously, intraperitoneally or topically
  • Parenteral administration refers to a mode of administration which includes intravenous, intramuscular, intraperitoneal, intrasternal, percutaneous and intramuscular injection and infusion.
  • the parenteral administration of the composition may be carried out by mixing the compound of the present invention with a pharmaceutically acceptable carrier under suitable purity, that is, preparing a unit dosage form by mixing the drug with a drug which is non-toxic to the receptor at the concentration and dosage to be used, Lt; / RTI >
  • the therapeutic agent of the present invention is administered in a therapeutically effective amount.
  • the therapeutically effective amount means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and includes the age, sex, weight, condition of the patient, symptoms of the disease, administration Time, and method of administration, and preferably 0.001 to 100 mg per day on an adult basis may be administered.
  • a cosmetic composition for improving dermatitis which comprises miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
  • a cosmetic composition for improving dermatitis which comprises, as an active ingredient, hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2.
  • the cosmetic composition of the present invention can inhibit the formation of MDC (macrophage-derived chemokine), suppress the expression of IgE and IL-4, and induce the expression of IFN-? As described above.
  • MDC macrophage-derived chemokine
  • an external preparation for skin for improving dermatitis comprising the composition according to an embodiment of the present invention.
  • the human obesity cell line (hmc-1) and PMA / IONOMYCIN are treated to cultivate human obesity cell line (hmc-1) induced inflammation-like activity.
  • microRNAs in the collected cells were selected by human microRNA array analysis.
  • the selected microRNAs were identified and analyzed by utilizing miRBASE, and miR-4507 microRNA shown in SEQ ID NO: 1 was synthesized from Biona (Korea Daejeon) Biotech Company and prepared for the test.
  • DNCB a skin disease inducer
  • atopic dermatosis was applied at a concentration of 1% per 100 ⁇ l for 1 week to induce atopic dermatosis.
  • miR-4507 microRNAs (1-10ug / mouse) were mixed in PBS and subcutaneously injected into experimental rats.
  • dermatitis symptoms were exacerbated in the mice to which the control group reagent was administered, and the treatment effect of dermatitis was shown in the experimental group to which miR-4507 microRNA was administered.
  • mice 5-week-old ICR mice were divided into two groups: control and experimental groups.
  • DNCB a skin disease inducer, was applied at a concentration of 1% per 100 ⁇ l for 1 week to induce atopic dermatosis. Subsequently, mice were subcutaneously injected with each microRNA (1-10 g / mouse) in PBS.
  • the serum of each mouse was extracted by orbital harvesting and ELISA experiments were performed to obtain serum concentrations of IL-4, IFN- ⁇ , MDC and IgE, which are indicators of atopic dermatitis.

Abstract

The present invention relates to microRNA having effects of preventing, treating, and ameliorating dermatitis and, more specifically, to a pharmaceutical composition containing miR-4507 microRNA as an active ingredient for prevention or treatment of dermatitis.

Description

MIR-4507 MICRORNA 유효 성분으로 함유하는 피부염의 예방 또는 치료용 약학 조성물MIR-4507 MICRORNA Pharmaceutical composition for prevention or treatment of dermatitis containing active ingredient
본 발명은 피부염의 개선에 효과를 갖는 작은 간섭 RNA에 관한 것으로, 더욱 구체적으로 miR-4507 microRNA를 유효성분으로 함유하는 피부염의 예방 또는 치료용 약학적 조성물에 관한 것이다.More particularly, the present invention relates to a pharmaceutical composition for preventing or treating dermatitis containing miR-4507 microRNA as an active ingredient.
아토피성 피부염은 현대인에게 빈번하게 발생되고 있는 의학적으로 원인이 불분명한 만성적 피부질환이다. 아토피성 피부염은 일반적으로 인간 면역체계의 불균형에 의해 초래된 질환으로 알려져 있다. 우리 몸의 면역은 B 림프구가 주관하는 체액성 면역과 T 림프구가 주관하는 세포성 면역으로 이루어져 있다. 세포성 면역을 주관하는 T 세포에는 Tc-세포와 Th-세포가 있으며, Th-세포는 Th-1과 Th-2로 구분된다. 상기 Th-1과 Th-2의 균형이 깨져 Th-2의 양이 갑자기 늘어나고 Th-1의 양이 줄어들면 알레르기 질환, 즉 아토피성 피부염, 알레르기성 비염, 알레르기성 천식 등의 질환이 발생하며, 반대로 Th-1의 양이 많이 늘어나게 되면 자가 면역 질환이 초래된다. 따라서 Th-2 면역반응의 과민은 아토피성 피부질환의 핵심요소이다. Th-2 면역반응은 즉시형 과민반응으로 IL-4, IL-5, IL-13 사이토카인의 분비를 매개로 이루어지며, IL-4와 IL-13은 B 세포의 isotype을 자극하고 IgE의 생성을 조절한다.Atopic dermatitis is a chronic skin disease that is medically undetermined and frequently occurring in modern humans. Atopic dermatitis is generally known as a disease caused by an imbalance of the human immune system. Our body's immune system consists of humoral immunity mediated by B lymphocytes and cellular immunity mediated by T lymphocytes. T-cells and Th-cells are involved in T-cells that mediate cell-mediated immunity, and Th-cells are divided into Th-1 and Th-2. When the amount of Th-2 is suddenly increased and the amount of Th-1 is decreased, the allergic diseases such as atopic dermatitis, allergic rhinitis and allergic asthma occur, Conversely, when the amount of Th-1 is increased, an autoimmune disease is caused. Thus, the hypersensitivity of the Th-2 immune response is a key component of atopic skin disease. The IL-4 and IL-13 stimulate the isotype of B cells and the production of IgE by the IL-4, IL-5 and IL-13 cytokines .
최근들어 국내외적으로 아토피성 피부염 발병이 소아에서 성인에 이르기까지 광범위하게 발생하고 있어 치료제의 개발이 시급하다. 현재 스테로이드 계열의 아토피성 피부염 억제제가 치료제로 사용되고 있으나 아토피성 피부염의 원천적인 해결방법이 아닌 일시적인 경감효과를 보여주며 동시에 내성이 생기는 부작용이 알려진바 적절하고 광범위한 치료제 개발이 필요한 실정이다.Recently, the incidence of atopic dermatitis has developed extensively from childhood to adulthood, so it is urgent to develop a therapeutic agent. Currently, steroid-based atopic dermatitis inhibitors are used as therapeutic agents. However, they are not a primary solution for atopic dermatitis but show a temporary relief effect. At the same time, side effects that cause resistance are known.
한편, CCL22는 T 세포에서 Th-2 phenotype의 CD+ T 세포들을 선택적으로 발현하는 CCR4의 리간드로써, 정상 환자군에 비해 아토피성 피부염 환자군에서 CCL22의 혈중 농도가 급격히 증가되어 있다. On the other hand, CCL22 is a ligand of CCR4 that selectively expresses CD2 + T cells of Th-2 phenotype in T cells. The concentration of CCL22 in the atopic dermatitis patients is rapidly increased compared with that of normal patients.
MDC는 아토피성 피부염 환자에서 그 혈청 농도가 현저히 증가한다고 보고된 케모카인으로서(Hijnen et. al., J. Allergy Clin. Immunol. 113(2) 334-340), 아토피성 피부염의 치료 물질로 사용되는 사이크로스포린 A나 코르티코스테로이드를 아토피성 피부염 환자에게 투여하였을 때는 MDC의 혈청 농도가 감소한다는 보고가 있으며(Y. Shimada et al., J. Dermatol. Sci., 34, 201-208, 2004), 또 사람 각질형성세포주인 HaCaT 세포에 hIFN-γ, TNF-α를 처리하였을 때 MDC가 다량 발현되는데 이러한 발현을 억제할 수 있는 물질은 아토피성 피부염 치료제로 사용될 수 있음이 제시된 바 있다 (Horikawa et. al., Int. Immunol., 14(7) 767-773, 2002).MDC is a chemokine that has been reported to significantly increase its serum concentration in patients with atopic dermatitis (Hijnen et al., J. Allergy Clin. Immunol. 113 (2) 334-340), used as a treatment for atopic dermatitis It has been reported that serum concentrations of MDC are decreased when ciclosporin A or corticosteroids are administered to patients with atopic dermatitis (Y. Shimada et al., J. Dermatol. Sci., 34, 201-208, 2004) In addition, when hIFN-y and TNF-a are treated with HaCaT cells, which are human keratinocyte cell lines, MDC is expressed in a large amount and a substance capable of inhibiting such expression can be used as a therapeutic agent for atopic dermatitis (Horikawa et. al., Int. Immunol., 14 (7) 767-773, 2002).
miRNA는 세포 내에 존재하는 20-25핵산(nucleotide) 길이의 작은 RNA (endogenous small RNA)의 일종으로 단백질을 합성하지 않는 DNA에서 유래되어 헤어핀-구조 전사체(hairpin-shaped transcript) 로 부터 생성이 된다. miRNA는 일반적으로 표적 mRNA의 3'-UTR의 상보적 서열에 결합하여 그 mRNA의 번역 억제 또는 불안정화를 유도하여, 궁극적으로 그 표적 mRNA의 단백질 합성을 억제하는 리프레서(repressor) 역할을 하게 된다. 하나의 miRNA는 여러 개의 mRNA를 타겟팅하며, mRNA 역시 여러 개의 miRNA에 의해 조절될 수 있다고 알려져 있다.miRNA is a kind of small RNA (endogenous small RNA) of 20-25 nucleotides in a cell, which is derived from a DNA that does not synthesize proteins and is generated from a hairpin-shaped transcript . miRNAs generally bind to the complementary sequence of the 3'-UTR of the target mRNA and induce the translation inhibition or destabilization of the mRNA, ultimately acting as a repressor to inhibit the protein synthesis of the target mRNA. One miRNA targets several mRNAs, and mRNA is also known to be regulated by several miRNAs.
현재 miRNA는 발달 시기, 세포사멸사, 지방 신진대사 및 조혈 세포 분화의 조절을 포함하는 여러 생물학적 과정에서 매우 중요한 역할을 하는 것으로 알려짐에 따라 생명과학분야에서 매우 큰 관심을 받고 있다. 하지만 암 등의 질병이나 발생 과정에서의 miRNA의 기능연구는 상당히 진행된 것에 비해 상대적으로 피부과학에서의 miRNA 역할에 대한 연구는 비교적 적은 상태이다.　Currently, miRNAs have been of great interest in life sciences as they are known to play a very important role in many biological processes including developmental timing, apoptosis, lipid metabolism, and regulation of hematopoietic differentiation. However, studies on the function of miRNAs in diseases such as cancer and development have progressed considerably, while studies on the role of miRNAs in dermatology have been relatively rare.
이에, 본 발명자들은 상기 종래기술들의 문제점들을 극복하기 위하여 예의 연구 노력한 결과, 21 ~ 25개 정도의 뉴클레오티드로 구성된 단일 염기 가닥인 microRNA를 피부염이 유발된 실험군에 피하 주사하는 경우, Th-2 면역반응의 지표인 IL-4의 발현을 감소시키고, Th-1 면역반응의 지표인 IFN-γ의 발현을 증가시키며, 또한 아토피 질환의 지표인 MDC의 생성을 억제하고, IgE의 면역반응을 급격히 감소시킬 수 있음을 확인하여 본 발명을 완성하게 되었다.The inventors of the present invention conducted extensive studies to overcome the problems of the prior art. As a result, when the microRNA, which is a single base strand composed of about 21 to 25 nucleotides, was subcutaneously injected into the dermatitis-induced experimental group, Th-2 immune response In addition, the expression of IFN-y, an indicator of Th-1 immune response, is suppressed, and the production of MDC, an indicator of atopic disease, is suppressed and the immune response of IgE is rapidly reduced The present invention has been completed.
본 발명의 주된 목적은 miR-4507 microRNA를 유효성분으로 함유하는 피부염 예방 또는 치료용 약학적 조성물을 제공하는데 있다.The main object of the present invention is to provide a pharmaceutical composition for preventing or treating dermatitis containing miR-4507 microRNA as an active ingredient.
본 발명의 또 다른 목적은 miR-4507 microRNA를 유효성분으로 함유하는 피부염 개선용 화장료 조성물을 제공하는데 있다. It is still another object of the present invention to provide a cosmetic composition for improving dermatitis containing miR-4507 microRNA as an active ingredient.
본 발명의 일 실시예에 따르면, 서열번호 1의 염기서열로 표시되는 miR-4507 microRNA를 유효성분으로 함유하는, 피부염의 예방 또는 치료용 약학적 조성물이 제공된다. According to one embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating dermatitis, which comprises miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
본 발명의 다른 일 실시예에 따르면, 서열번호 2의 염기서열로 표시되는 hsa-mir-4507 microRNA를 유효성분으로 함유하는, 피부염의 예방 또는 치료용 약학적 조성물이 제공된다.According to another embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating dermatitis, which comprises hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2 as an active ingredient.
일 측에 따르면, 상기 조성물은 MDC(macrophage-derived chemokine)의 생성을 억제할 수 있다.According to one aspect, the composition can inhibit the production of macrophage-derived chemokines (MDCs).
일 측에 따르면, 상기 조성물은 IgE의 발현을 억제할 수 있다.According to one aspect, the composition can inhibit the expression of IgE.
일 측에 따르면, 상기 조성물은 IL-4의 발현을 억제할 수 있다.According to one aspect, the composition can inhibit the expression of IL-4.
일 측에 따르면, 상기 조성물은 IFN- γ의 발현을 유도할 수 있다.According to one aspect, the composition can induce the expression of IFN- ?.
본 발명의 또 다른 일 실시예에 따르면, 서열번호 1의 염기서열로 표시되는 miR-4507 microRNA를 유효성분으로 함유하는, 피부염의 개선용 화장료 조성물이 제공된다. According to another embodiment of the present invention, there is provided a cosmetic composition for improving dermatitis, which comprises miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
본 발명의 또 다른 일 실시예에 따르면, 서열번호 2의 염기서열로 표시되는 hsa-mir-4507 microRNA를 유효성분으로 함유하는, 피부염의 개선용 화장료 조성물이 제공된다.According to another embodiment of the present invention, there is provided a cosmetic composition for improving dermatitis, which comprises, as an active ingredient, hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2.
일 측에 따르면, 상기 조성물은 IgE의 발현을 억제할 수 있다.According to one aspect, the composition can inhibit the expression of IgE.
일 측에 따르면, 상기 조성물은 IL-4의 발현을 억제할 수 있다.According to one aspect, the composition can inhibit the expression of IL-4.
일 측에 따르면, 상기 조성물은 IFN- γ의 발현을 유도할 수 있다.According to one aspect, the composition can induce the expression of IFN- ?.
본 발명의 또 다른 일 실시예에 따르면, 본 발명의 일 실시예에 따른 조성물을 포함하는, 피부염 개선용 피부 외용제가 제공된다.According to another embodiment of the present invention, there is provided an external preparation for skin for improving dermatitis comprising the composition according to an embodiment of the present invention.
일 측에 따르면, 상기 피부 외용제는, 겔, 크림, 유액, 화장수, 팩, 로션, 에센스, 파우더, 비누, 입욕제 및 스프레이로 이루어진 군에서 선택되는 제형을 가질 수 있다.According to one aspect, the external preparation for skin may have a formulation selected from the group consisting of a gel, a cream, a milky lotion, a lotion, a pack, a lotion, an essence, a powder, a soap, a bath agent and a spray.
본 발명의 피부염의 예방 또는 치료용 약학적 조성물 및 피부염의 개선용 화장료 조성물은 Th-2 면역반응의 지표인 IL-4의 발현을 감소시키고, Th-1 면역반응의 지표인 IFN-γ의 발현을 증가시키며, 아토피 질환의 지표인 MDC의 생성을 억제하고 IgE의 면역반응을 급격히 감소시킬 수 있다. 또한, 피부염의 대표적인 증상 중 하나인 가려움증을 완화시킬 수 있다.The pharmaceutical composition for the prevention or treatment of dermatitis of the present invention and the cosmetic composition for improving dermatitis reduce the expression of IL-4, which is an index of Th-2 immune response, and the expression of IFN-y, , Inhibiting the production of MDC, an indicator of atopic disease, and rapidly reducing the immune response of IgE. In addition, itching can be relieved, which is one of the typical symptoms of dermatitis.
또한, miR-4507 microRNA는 인간유래의 RNA로, 부작용에 대한 위험성이 낮고 안전한 물질이므로 이를 유효성분으로 함유하는 본 발명의 조성물은 피부 외용제나 화장품으로 유용하게 이용될 수 있다. In addition, since miR-4507 microRNA is a human-derived RNA, it is a low-risk and safe substance for side effects, and thus the composition of the present invention containing it as an active ingredient can be usefully used as an external preparation for skin or cosmetics.
도 1은 miR-4507 microRNA를 함유하는 조성물이 피부염을 유도한 마우스에서 피부의 염증 완화 효과를 가지는 것을 보여주는 도면이다.FIG. 1 is a view showing that a composition containing miR-4507 microRNA has an inflammation-reducing effect on the skin in a dermatitis-induced mouse.
도 2는 miR-4507 microRNA를 함유하는 조성물이 피부염을 유도시킨 마우스에서 Th2 cell을 유도하는 케모카인인 MDC를 억제하는 결과를 보여주는 도면이다.FIG. 2 is a graph showing a result of inhibiting MDC, a chemokine that induces Th2 cells in a dermatitis-induced mouse, wherein the composition containing miR-4507 microRNA is inhibited.
도 3은 miR-4507 microRNA를 함유하는 조성물이 피부염증 마우스의 가려움증을 억제한 효과를 보여주는 도면이다. Fig. 3 shows the effect of a composition containing miR-4507 microRNA to suppress itching in skin inflammation mice.
도 4는 miR-4507 microRNA를 함유하는 조성물이 피부염의 유발 항체인 IgE의 발현을 억제한 결과를 나타낸 도면이다.FIG. 4 is a graph showing the results of inhibiting the expression of IgE, a dermatitis-inducing antibody, in a composition containing miR-4507 microRNA.
도 5는 miR-4507 microRNA를 함유하는 조성물이 피부염의 유발 사이토카인인 IL-4의 발현을 억제한 결과를 나타낸 도면이다.FIG. 5 is a graph showing the results of inhibiting the expression of IL-4, a dermatitis-inducing cytokine, in a composition containing miR-4507 microRNA.
도6은 miR-4507 microRNA를 함유하는 조성물이 피부염의 관련 IFN-gamma 발현을 유도한 결과를 나타낸 도면이다.FIG. 6 is a graph showing a result of induction of a related IFN-gamma expression of dermatitis by a composition containing miR-4507 microRNA.
이하, 실시예를 통하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .
실시예에서 사용한 용어는 단지 특정한 실시예를 설명하기 위해 사용된 것으로, 실시예를 한정하려는 의도가 아니다. 단수의 표현은 문맥상 명백하게 다르게 뜻하지 않는 한, 복수의 표현을 포함한다. 본 명세서에서, "포함하다" 또는 "가지다" 등의 용어는 명세서 상에 기재된 특징, 숫자, 단계, 동작, 구성 요소, 부품 또는 이들을 조합한 것이 존재함을 지정하려는 것이지, 하나 또는 그 이상의 다른 특징들이나 숫자, 단계, 동작, 구성 요소, 부품 또는 이들을 조합한 것들의 존재 또는 부가 가능성을 미리 배제하지 않는 것으로 이해되어야 한다.The terms used in the examples are used only to illustrate specific embodiments and are not intended to limit the embodiments. The singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, the terms "comprises" or "having" and the like refer to the presence of stated features, integers, steps, operations, elements, components, or combinations thereof, But do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, or combinations thereof.
다르게 정의되지 않는 한, 기술적이거나 과학적인 용어를 포함해서 여기서 사용되는 모든 용어들은 실시예가 속하는 기술 분야에서 통상의 지식을 가진 자에 의해 일반적으로 이해되는 것과 동일한 의미를 가지고 있다. 일반적으로 사용되는 사전에 정의되어 있는 것과 같은 용어들은 관련 기술의 문맥 상 가지는 의미와 일치하는 의미를 가지는 것으로 해석되어야 하며, 본 출원에서 명백하게 정의하지 않는 한, 이상적이거나 과도하게 형식적인 의미로 해석되지 않는다.Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this embodiment belongs. Terms such as those defined in commonly used dictionaries are to be interpreted as having a meaning consistent with the contextual meaning of the related art and are to be interpreted as either ideal or overly formal in the sense of the present application Do not.
또한, 첨부 도면을 참조하여 설명함에 있어, 도면 부호에 관계없이 동일한 구성 요소는 동일한 참조 부호를 부여하고 이에 대한 중복되는 설명은 생략하기로 한다. 실시예를 설명함에 있어서 관련된 공지 기술에 대한 구체적인 설명이 실시예의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우 그 상세한 설명을 생략한다.In the following description of the present invention with reference to the accompanying drawings, the same components are denoted by the same reference numerals regardless of the reference numerals, and redundant explanations thereof will be omitted. In the following description of the embodiments, a detailed description of related arts will be omitted if it is determined that the gist of the embodiments may be unnecessarily blurred.
본 발명의 일 실시예에 따르면, 서열번호 1의 염기서열로 표시되는 miR-4507 microRNA를 유효성분으로 함유하는, 피부염의 예방 또는 치료용 약학적 조성물이 제공된다.According to one embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating dermatitis, which comprises miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
또한, 본 발명의 다른 일 실시예에 따르면, 서열번호 2의 염기서열로 표시되는 hsa-mir-4507 microRNA를 유효성분으로 함유하는, 피부염의 예방 또는 치료용 약학적 조성물이 제공된다.According to another embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating dermatitis, which comprises hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2 as an active ingredient.
본 발명의 조성물에 함유되는 상기 microRNA는 21 뉴클레오티드의 miR-4507 microRNA의 염기서열로써, 바람직하게는 하기 표 1에 나타낸 서열번호 1 또는 서열번호 2의 microRNA일 수 있다.The microRNA contained in the composition of the present invention is a nucleotide sequence of miR-4507 microRNA of 21 nucleotides, preferably microRNA of SEQ ID NO: 1 or SEQ ID NO: 2 shown in Table 1 below.
상기 조성물은 MDC(macrophage-derived chemokine)의 생성을 억제하고, IgE와 IL-4의 발현을 억제할 수 있으며, IFN- γ의 발현을 유도할 수 있다.The composition inhibits the formation of MDC (macrophage-derived chemokine), inhibits the expression of IgE and IL-4, and can induce the expression of IFN- ?.
Figure PCTKR2018007562-appb-I000001
Figure PCTKR2018007562-appb-I000001
본 발명에 따른 microRNA 폴리뉴클레오티드 서열은 발현 조절 서열에 작동 가능하게 연결될 수 있으며, 작동 가능하게 연결된 유전자 서열과 발현 조절 서열은 선택 마커 및 복제 개시점(replication origin)을 같이 포함하고 있는 하나의 발현벡터 내에 포함될 수 있다. A microRNA polynucleotide sequence according to the present invention may be operably linked to an expression control sequence, wherein the operably linked gene sequence and expression control sequence are an expression vector containing a selectable marker and a replication origin Lt; / RTI >
"작동 가능하게 연결(operably linked)" 된다는 것은 적절한 분자가 발현 조절 서열에 결합될 때 유전자 발현을 가능하게 하는 방식으로 연결된 유전자 및 발현 조절 서열일 수 있다. &Quot; Operably linked " may be a gene and expression control sequence linked in such a way as to enable gene expression when the appropriate molecule is coupled to the expression control sequence.
"발현 조절 서열(expression control sequence)"이란 특정한 숙주 세포에서 작동 가능하게 연결된 폴리뉴클레오티드 서열의 발현을 조절하는 DNA 서열을 의미한다. 그러한 조절 서열은 전사를 실시하기위한 프로모터, 전사를 조절하기 위한 임의의 오퍼레이터 서열, 적합한 mRNA 리보좀 결합 부위를 코딩하는 서열, 전사 및 해독의 종결을 조절하는 서열을 포함한다.&Quot; Expression control sequence " means a DNA sequence that regulates the expression of a polynucleotide sequence operably linked to a particular host cell. Such regulatory sequences include promoters for conducting transcription, any operator sequences for regulating transcription, sequences encoding suitable mRNA ribosome binding sites, sequences controlling the termination of transcription and translation.
본 발명의 피부염 예방, 치료 효과를 유도하는 miR-4507 microRNA는 통상 사용되는 유전자합성에 의해 제조될 수 있으며, 제조된 microRNA를 체내에 도입시킬 때, 상기 microRNA는 바이러스벡터나 플라스미드 벡터 등에 의해서도 발현될 수 있다.MiR-4507 microRNA, which induces the dermatitis prevention and therapeutic effect of the present invention, can be produced by commonly used gene synthesis, and when the produced microRNA is introduced into the body, the miRNA is also expressed by a viral vector or a plasmid vector .
본 발명의 약학적 조성물은 약제로 이용되기 위해서 약제학적 분야에서 공지된 방법에 의해 제조될 수 있으며, 약학적으로 허용되는 담체, 부형제, 희석제, 안정화제, 방부제 등과 혼합하여 분말, 과립, 정제, 캡슐제, 또는 주사제 등의 제형으로 제조되어 사용될 수 있다. The pharmaceutical composition of the present invention may be prepared by a method known in the pharmaceutical art to be used as a pharmaceutical agent and may be prepared by mixing with a pharmaceutically acceptable carrier, an excipient, a diluent, a stabilizer, Capsules, injections, and the like.
상기 희석제로는 프로필렌 글리콜, 폴리에틸렌 글리콜, 올리브유, 땅콩유와 같은 식물성유 등의 비수성 용매나 염수(바람직하게는 0.8%의 염수), 완충 매질을 포함한 물(바람직하게는 0.05M의 인산염 완충액) 등의 수성 용매 등을 들 수 있으나, 이에 한정되는 것은 아니다.The diluent may be a non-aqueous solvent such as propylene glycol, polyethylene glycol, vegetable oil such as olive oil or peanut oil, or water (preferably 0.05 M phosphate buffer) containing saline (preferably 0.8% saline) , And the like, but the present invention is not limited thereto.
상기 부형제로는 전분, 글루코스, 락토스, 수크로스, 젤라틴, 맥아, 쌀, 밀가루, 백악, 실리카 겔, 나트륨 스테아레이트, 글리세롤 모노스테아레이트, 활석, 나트륨 클로라이드, 무수 탈지유, 글리세롤, 프로필렌, 글리콜, 물, 에탄올 등을 들 수 있으나, 이에 한정되는 것은 아니다.Examples of the excipient include starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, anhydrous skimmed milk, glycerol, , Ethanol, and the like, but are not limited thereto.
상기 안정화제로는 소르비톨, 만니톨, 전분, 수크로스, 덱스트란, 글루타메이트, 글루코스 등의 탄수화물이나 분유, 혈청 알부민, 카제인 등의 동물성, 식물성 또는 미생물성 단백질 등의 단백질을 들 수 있으나, 이에 한정되는 것은 아니다.Examples of the stabilizer include proteins such as carbohydrates such as sorbitol, mannitol, starch, sucrose, dextran, glutamate and glucose and animal, vegetable or microbial proteins such as milk powder, serum albumin and casein. no.
상기 방부제로는 티메로살, 메르티올레이트, 젠타마이신, 네오마이신, 니스타틴, 암포테리신 B, 테트라사이클린, 페니실린, 스트렙토마이신, 폴리믹신 B 등을 들 수 있으나, 이에 한정되는 것은 아니다.Examples of the preservative include, but are not limited to, thimerosal, mertiolate, gentamicin, neomycin, nystatin, amphotericin B, tetracycline, penicillin, streptomycin, polyamicin B and the like.
상기 약학적 조성물은 개별 치료제로 투여되거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와 순차로 또는 동시에 투여될 수 있다. 그리고 단일 또는 다중 투여될 수 있다. 상기 요소를 모두 고려하여 부작용없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The pharmaceutical composition may be administered as an individual therapeutic agent or in combination with another therapeutic agent, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by a person skilled in the art.
또한, 이들은 비경구 투여(예컨대, 정맥 내, 피하, 복강 내 또는 국소에 적용)하거나 경구 투여될 수 있다. 비경구 투여는 정맥내, 근육내, 복강내, 흉골내, 경피 및 동맥내 주사 및 주입을 포함하는 투여 방식을 의미한다. 상기 조성물의 비경구 투여는 바람직한 순도 하에 약제학적으로 허용 가능한 담체, 즉 사용되는 농도와 투여량에서 수용체에 비독성이고 다른 제제 성분과 혼화 될 수 있는 것을 혼합하여 단위 투여량의 제형으로 조제하는 것이 바람직할 수 있다.They may also be administered parenterally (e. G., Intravenously, subcutaneously, intraperitoneally or topically) or orally. Parenteral administration refers to a mode of administration which includes intravenous, intramuscular, intraperitoneal, intrasternal, percutaneous and intramuscular injection and infusion. The parenteral administration of the composition may be carried out by mixing the compound of the present invention with a pharmaceutically acceptable carrier under suitable purity, that is, preparing a unit dosage form by mixing the drug with a drug which is non-toxic to the receptor at the concentration and dosage to be used, Lt; / RTI >
본 발명의 치료제는 치료학적으로 유효한 양으로 투여한다. 상기 치료학적으로 유효한 양(therapeutically effective amount)은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 환자의 연령, 성별, 체중, 건강상태, 질병의 증상, 투여시간, 투여방법에 따라 적적히 선택될 수 있으며, 바람직하게는 성인기준 1일 0.001 ~ 100 mg이 투여될 수 있다.The therapeutic agent of the present invention is administered in a therapeutically effective amount. The therapeutically effective amount means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and includes the age, sex, weight, condition of the patient, symptoms of the disease, administration Time, and method of administration, and preferably 0.001 to 100 mg per day on an adult basis may be administered.
본 발명의 또 다른 일 실시예에 따르면, 서열번호 1의 염기서열로 표시되는 miR-4507 microRNA를 유효성분으로 함유하는, 피부염의 개선용 화장료 조성물이 제공된다. According to another embodiment of the present invention, there is provided a cosmetic composition for improving dermatitis, which comprises miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
또한, 본 발명의 또 다른 일 실시예에 따르면, 서열번호 2의 염기서열로 표시되는 hsa-mir-4507 microRNA를 유효성분으로 함유하는, 피부염의 개선용 화장료 조성물이 제공된다.According to still another embodiment of the present invention, there is provided a cosmetic composition for improving dermatitis, which comprises, as an active ingredient, hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2.
본 발명의 화장료 조성물은 전술한 바와 같이 MDC(macrophage-derived chemokine)의 생성을 억제하고, IgE와 IL-4의 발현을 억제할 수 있으며, IFN- γ의 발현을 유도할 수 있다.The cosmetic composition of the present invention can inhibit the formation of MDC (macrophage-derived chemokine), suppress the expression of IgE and IL-4, and induce the expression of IFN-? As described above.
본 발명의 또 다른 일 실시예에 따르면, 본 발명의 일 실시예에 따른 조성물을 포함하는, 피부염 개선용 피부 외용제가 제공된다.According to another embodiment of the present invention, there is provided an external preparation for skin for improving dermatitis comprising the composition according to an embodiment of the present invention.
또한, 본 발명의 피부 외용제는, 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어 겔, 크림, 유액, 화장수, 팩, 로션, 에센스, 파우더, 비누, 입욕제 및 스프레이로 이루어진 군에서 선택되는 제형을 가질 수 있다.In addition, the external preparation for skin of the present invention can be prepared in any formulations conventionally produced in the art, and can be prepared, for example, as gels, creams, milks, lotions, packs, lotions, essences, powders, soaps, Lt; RTI ID = 0.0 > a < / RTI >
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.
<실시예 1. miR-4507 microRNA의 선별 및 제작> & Lt; Example 1 : Selection and production of miR-4507 microRNA >
인체 비만세포주(hmc-1)와 PMA/IONOMYCIN을 처리하여 염증 유사 활성이 유도된 인체 비만세포주(hmc-1)를 배양한다. 배양된 세포주 107 세포를 수득하고, 세포내 microRNA를 microRNA Isolation kit(quiagen, germany)를 이용하여 수집하였다. The human obesity cell line (hmc-1) and PMA / IONOMYCIN are treated to cultivate human obesity cell line (hmc-1) induced inflammation-like activity. Obtaining a cultured cell line 10 7 cells, and the cells were collected microRNA using microRNA Isolation kit (quiagen, germany).
수집된 세포내 microRNA들을 human microRNA array 분석을 통해서 염증에 관련된 microRNA를 선별하였다. 선별된 microRNA는 miRBASE 를 활용하여 서열을 확인 분석하여, 서열번호 1로 표시되는 miR-4507 microRNA를 바이오니어(한국 대전)바이오텍 회사로부터 합성하여 시험에 준비하였다. The microRNAs in the collected cells were selected by human microRNA array analysis. The selected microRNAs were identified and analyzed by utilizing miRBASE, and miR-4507 microRNA shown in SEQ ID NO: 1 was synthesized from Biona (Korea Daejeon) Biotech Company and prepared for the test.
<실시예 2. 피부염 모델의 제조 및 치료 효과 평가>&Lt; Example 2 &gt; Preparation and treatment efficacy of dermatitis model>
5주령된 ICR mouse를 5 마리씩 대조군과 실험군의 두 그룹으로 나눈 후, 각각의 쥐에 피부병 유발 물질인 DNCB를 1%의 농도로 1주일간 100 μl씩 도포하여 아토피와 유사한 피부병을 유발시켰다. 이후, 실험군 쥐에 miR-4507 microRNA를 (1-10ug/mouse)의 농도로 PBS에 혼합하여 피하 주사 하고, 대조군 쥐에는 PBS만을 주사하였다. 그 결과, 도 1에 나타낸 바와 같이, 대조군 시약을 투여한 쥐에서는 피부염 증상이 악화되었고, miR-4507 microRNA를 투여한 실험군에서는 피부염의 치료 효과가 나타남을 확인할 수 있다.5-week-old ICR mice were divided into two groups: control and experimental groups. DNCB, a skin disease inducer, was applied at a concentration of 1% per 100 μl for 1 week to induce atopic dermatosis. Subsequently, miR-4507 microRNAs (1-10ug / mouse) were mixed in PBS and subcutaneously injected into experimental rats. As a result, as shown in Fig. 1, dermatitis symptoms were exacerbated in the mice to which the control group reagent was administered, and the treatment effect of dermatitis was shown in the experimental group to which miR-4507 microRNA was administered.
<실시예 3. 아토피성 피부염 모델 제조 및 아토피성 피부염 지표의 측정> &Lt; Example 3: Preparation of atopic dermatitis model and measurement of atopic dermatitis index >
5주령된 ICR mouse를 5 마리씩 대조군과 실험군의 두 그룹으로 나눈 후, 각각의 쥐에 피부병 유발 물질인 DNCB를 1%의 농도로 1주일간 100 μl씩 도포하여 아토피와 유사한 피부병을 유발시켰다. 이후, 실험군 쥐에 각각의 microRNA를 (1-10ug/mouse)의 농도로 PBS에 혼합하여 피하 주사 하였다. 5-week-old ICR mice were divided into two groups: control and experimental groups. DNCB, a skin disease inducer, was applied at a concentration of 1% per 100 μl for 1 week to induce atopic dermatosis. Subsequently, mice were subcutaneously injected with each microRNA (1-10 g / mouse) in PBS.
투여한지 일주일이 되는 시점에 안와 채취법을 이용하여 각각의 쥐의 혈청을 뽑아 아토피성 피부염의 지표인 IL-4, IFN-γ, MDC, IgE의 혈청내 농도를 구하기 위해 ELISA 실험을 수행하였다. At one week after administration, the serum of each mouse was extracted by orbital harvesting and ELISA experiments were performed to obtain serum concentrations of IL-4, IFN-γ, MDC and IgE, which are indicators of atopic dermatitis.
실험 결과, miR-4507 microRNA를 피부염이 유발된 쥐에 투여한 경우, MDC 생성과 IL-4의 발현이 억제되고(도 2 및 도 5 참조), 아토피 질환의 지표인 IgE 면역반응이 급격히 감소하였으며(도 4 참조), IFN-γ의 발현이 유도되는 것(도 6 참조)을 확인할 수 있었다.As a result, when MDR-4507 microRNA was administered to dermatitis-induced rats, MDC production and IL-4 expression were inhibited (see FIGS. 2 and 5), and IgE immune response, which is an indicator of atopic disease, (See FIG. 4), and IFN-? Expression was induced (see FIG. 6).
또한, 도 3에 나타낸 바와 같이, miR-4507 microRNA를 투여한 실험군의 경우 피부염 증상 중 하나인 가려움증이 눈에 띄게 감소함을 확인할 수 있었다.In addition, as shown in Fig. 3, it was confirmed that the test group to which miR-4507 microRNA was administered markedly decreased the itching, one of the symptoms of dermatitis.

Claims (14)

  1. 서열번호 1의 염기서열로 표시되는 miR-4507 microRNA를 유효성분으로 함유하는, 피부염의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating dermatitis, comprising miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
  2. 서열번호 2의 염기서열로 표시되는 hsa-mir-4507 microRNA를 유효성분으로 함유하는, 피부염의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating dermatitis, which comprises hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2 as an active ingredient.
  3. 제1항 또는 제2항에 있어서,3. The method according to claim 1 or 2,
    상기 조성물은 MDC(macrophage-derived chemokine)의 생성을 억제하는, 피부염의 예방 또는 치료용 약학적 조성물.Wherein said composition inhibits the production of MDC (macrophage-derived chemokine).
  4. 제1항 또는 제2항에 있어서,3. The method according to claim 1 or 2,
    상기 조성물은 IgE의 발현을 억제하는, 피부염의 예방 또는 치료용 약학적 조성물.The pharmaceutical composition for preventing or treating dermatitis, wherein the composition inhibits the expression of IgE.
  5. 제1항 또는 제2항에 있어서,3. The method according to claim 1 or 2,
    상기 조성물은 IL-4의 발현을 억제하는, 피부염의 예방 또는 치료용 약학적 조성물.Wherein said composition inhibits the expression of IL-4.
  6. 제1항 또는 제2항에 있어서,3. The method according to claim 1 or 2,
    상기 조성물은 IFN-γ의 발현을 유도하는, 피부염의 예방 또는 치료용 약학적 조성물.Wherein said composition induces the expression of IFN-y.
  7. 서열번호 1의 염기서열로 표시되는 miR-4507 microRNA를 유효성분으로 함유하는, 피부염의 개선용 화장료 조성물.A cosmetic composition for improving dermatitis, comprising miR-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 1 as an active ingredient.
  8. 서열번호 2의 염기서열로 표시되는 hsa-mir-4507 microRNA를 유효성분으로 함유하는, 피부염의 개선용 화장료 조성물.A cosmetic composition for improving dermatitis comprising hsa-mir-4507 microRNA represented by the nucleotide sequence of SEQ ID NO: 2 as an active ingredient.
  9. 제7항 또는 제8항에 있어서,9. The method according to claim 7 or 8,
    상기 조성물은 MDC(macrophage-derived chemokine)의 생성을 억제하는, 피부염의 개선용 화장료 조성물.Wherein said composition inhibits the production of MDC (macrophage-derived chemokine).
  10. 제7항 또는 제8항에 있어서,9. The method according to claim 7 or 8,
    상기 조성물은 IgE의 발현을 억제하는, 피부염의 개선용 화장료 조성물.Wherein the composition inhibits the expression of IgE.
  11. 제7항 또는 제8항에 있어서,9. The method according to claim 7 or 8,
    상기 조성물은 IL-4의 발현을 억제하는, 피부염의 개선용 화장료 조성물.Wherein said composition inhibits the expression of IL-4.
  12. 제7항 또는 제8항에 있어서,9. The method according to claim 7 or 8,
    상기 조성물은 IFN-γ의 발현을 유도하는, 피부염의 개선용 화장료 조성물.Wherein said composition induces expression of IFN-y.
  13. 제7항 또는 제8항의 조성물을 포함하는, 피부염 개선용 피부 외용제.8. An external preparation for skin for improving dermatitis comprising the composition of claim 7 or 8.
  14. 제13항에 있어서,14. The method of claim 13,
    상기 피부 외용제는, 겔, 크림, 유액, 화장수, 팩, 로션, 에센스, 파우더, 비누, 입욕제 및 스프레이로 이루어진 군에서 선택되는 제형을 가지는, 피부염 개선용 피부 외용제.The external preparation for skin for improving dermatitis has a formulation selected from the group consisting of gel, cream, milky lotion, lotion, pack, lotion, essence, powder, soap, bath agent and spray.
PCT/KR2018/007562 2017-07-05 2018-07-04 Pharmaceutical composition containing mir-4507 microrna as active ingredient for prevention or treatment of dermatitis WO2019009606A1 (en)

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