KR20200098255A - Composition for preventing and treating arthritis comprising a DEC1 expression and activity modulator - Google Patents
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Abstract
Description
본 발명은 골관절염 예방 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for preventing or treating osteoarthritis.
골관절염(osteoarthritis, OA)이란 퇴행성 관절염이라고도 불리며, 관절을 보호하고 있는 연골의 점진적인 손상이나 퇴행성 변화로 인해 관절을 이루는 뼈와 인대 등에 손상이 일어나서 염증과 통증이 생기는 질환이다. Osteoarthritis (OA), also called degenerative arthritis, is a disease in which inflammation and pain occur due to damage to the bones and ligaments that make up the joint due to gradual damage or degenerative changes in the cartilage that protects the joint.
연골 조직은 연골 세포만으로 이루어진 조직으로서, 관절 내 정상적 연골세포는 대사적 활성이 있고 증식 및 연골기질분자의 합성이나 분해 등이 매우 느리게 일어나는 특징을 가진다. 또한, 연골 조직은 혈관과 신경 조직이 없어 영양과 산소 공급이 원활하지 않기 때문에 손상 부위의 재생이 느리다. 따라서, 현재까지 골관절염은 인공관절 치환법 외에 근본적인 치료법이 확립되지 않은 질병이다.Cartilage tissue is a tissue consisting of only cartilage cells, and normal cartilage cells in a joint have metabolic activity, and proliferation and synthesis or decomposition of cartilage matrix molecules occur very slowly. In addition, since cartilage tissue does not have blood vessels and nerve tissues, nutrition and oxygen supply is not smooth, regeneration of the damaged area is slow. Therefore, until now, osteoarthritis is a disease for which no fundamental treatment has been established other than artificial joint replacement.
한편, 생체주기 조절인자 발현의 증감은 노화에 따라 그 폭이 감소하여 대사과정의 변화를 야기하고, 조직의 성장과 복구가 감소하여 노화에 따를 퇴행성 질환을 야기하는 것으로 여겨진다. 연골 조직에서 발현하는 유전자 중 약 3.9%에 달하는 유전자의 발현이 생체주기에 따라 조절되는 것으로 알려져 있으며, 이는 연골기질 분해 및 연골 퇴행이 생체주기에 따라 조절될 수 있음을 암시한다.On the other hand, the increase or decrease in the expression of a life cycle regulator decreases with aging, causing a change in metabolic processes, and decreases in tissue growth and repair, and is believed to cause degenerative diseases following aging. It is known that the expression of about 3.9% of the genes expressed in cartilage tissues is regulated according to the life cycle, suggesting that cartilage matrix degradation and cartilage degeneration can be regulated according to the life cycle.
다만, 아직까지 골관절염에 대해 생체리듬 조절인자의 기능과 역할을 규명한 연구는 없는 실정이다.However, there are no studies yet to investigate the function and role of biological rhythm regulators in osteoarthritis.
본 발명자들은 골관절염 예방 또는 치료 효과를 나타내는 생물-화학적인 물질을 찾고자 노력하였다. 그 결과, DEC1의 발현 억제 시 연골기질 분해효소 및 연골퇴행 유도 인자들의 발현이 감소함을 규명함으로써, 본 발명을 완성하게 되었다.The present inventors have tried to find a bio-chemical substance exhibiting an effect of preventing or treating osteoarthritis. As a result, it was found that the expression of cartilage matrix degrading enzyme and cartilage degeneration inducing factors decreased when the expression of DEC1 was suppressed, thereby completing the present invention.
따라서, 본 발명의 목적은 DEC1(Differentiated embryo chondrocyte expressed gene 1)의 발현 또는 활성 억제제를 유효성분으로 포함하는 골관절염의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating osteoarthritis comprising an inhibitor of expression or activity of DEC1 (Differentiated embryo chondrocyte expressed gene 1) as an active ingredient.
본 발명의 다른 목적은 DEC1(Differentiated embryo chondrocyte expressed gene 1)의 발현 또는 활성 수준을 측정하는 제제를 포함하는 골관절염 진단용 조성물을 제공하는 것이다.Another object of the present invention is to provide a composition for diagnosing osteoarthritis comprising an agent for measuring the expression or activity level of DEC1 (Differentiated embryo chondrocyte expressed gene 1).
본 발명의 또 다른 목적은 골관절염 진단을 위한 정보제공방법을 제공하는 것이다.Another object of the present invention is to provide a method of providing information for diagnosing osteoarthritis.
본 발명의 또 다른 목적은 골관절염 치료제의 스크리닝 방법을 제공하는 것이다.Another object of the present invention is to provide a screening method for a therapeutic agent for osteoarthritis.
본 발명자들은 골관절염 예방 또는 치료 효과를 나타내는 생물-화학적인 물질을 찾고자 노력하였다. 그 결과, DEC1의 발현 억제 시 연골기질 분해효소 및 연골퇴행 유도 인자들의 발현이 감소함을 규명하였다.The present inventors have tried to find a bio-chemical substance exhibiting an effect of preventing or treating osteoarthritis. As a result, it was found that when the expression of DEC1 was suppressed, the expression of cartilage matrix degrading enzyme and cartilage degeneration inducing factors decreased.
본 발명은 DEC1의 발현 또는 활성 억제제를 유효성분으로 포함하는 골관절염의 예방 또는 치료용 약학적 조성물 및 DEC1의 발현 또는 활성 수준을 측정하는 제제를 포함하는 골관절염 진단용 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for prophylaxis or treatment of osteoarthritis comprising an inhibitor of DEC1 expression or activity as an active ingredient, and a composition for diagnosing osteoarthritis comprising an agent for measuring the expression or activity level of DEC1.
이하, 본 발명을 더욱 자세히 설명하고자 한다.Hereinafter, the present invention will be described in more detail.
본 발명의 일 양태는 DEC1(Differentiated embryo chondrocyte expressed gene 1)의 발현 또는 활성 억제제를 유효성분으로 포함하는 골관절염의 예방 또는 치료용 약학적 조성물에 관한 것이다.One aspect of the present invention relates to a pharmaceutical composition for preventing or treating osteoarthritis comprising an inhibitor of expression or activity of DEC1 (Differentiated embryo chondrocyte expressed gene 1) as an active ingredient.
본 발명에서 사용되는 용어, "예방"이란 본 발명에 따른 약학적 조성물의 투여에 의해 골관절염을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.As used herein, the term "prevention" refers to any action that suppresses osteoarthritis or delays the onset of osteoarthritis by administration of the pharmaceutical composition according to the present invention.
본 발명에서 사용되는 용어, "치료"란 본 발명에 따른 약학적 조성물의 투여에 의해 골관절염에 대한 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다.The term "treatment" used in the present invention refers to any action in which symptoms for osteoarthritis are improved or advantageously changed by administration of the pharmaceutical composition according to the present invention.
본 발명의 조성물에 의한 예방, 치료 대상 질병인 골관절염은 퇴행성 관절염이라고도 불리며, 관절을 보호하고 있는 연골의 점진적인 손상이나 퇴행성 변화로 인해 관절을 이루는 뼈와 인대 등에 손상이 일어나서 염증과 통증이 생기는 질환을 의미한다. 본 발명의 조성물은 유전자 표적 치료의 일환으로서, DEC1을 표적으로 하는 골관절염 치료제를 제공한다.Osteoarthritis, which is a disease to be prevented or treated by the composition of the present invention, is also called degenerative arthritis, and it is a disease that causes inflammation and pain due to damage to the bones and ligaments that make up the joint due to gradual damage or degenerative changes in the cartilage protecting the joint. it means. The composition of the present invention provides a therapeutic agent for osteoarthritis targeting DEC1 as part of gene targeted therapy.
상기 DEC1(Differentiated embryo chondrocyte expressed gene 1) 유전자는 생체리듬의 핵심 조절인자인 조절인자인 BMAL1(Arntl aryl hydrocarbon receptor nuclear translocator-like) 및 CLOCK(Circadian locomotor output cycles kaput) 이형이량체의 활성을 조절하여 생체리듬을 조절하는 전사체로 알려져 있으나, DEC1의 발현 또는 활성 억제를 통한 골관절염 치료에 대해서는 아직 보고된 바가 없다.The DEC1 (Differentiated embryo chondrocyte expressed gene 1) gene regulates the activity of BMAL1 (Arntl aryl hydrocarbon receptor nuclear translocator-like) and CLOCK (Circadian locomotor output cycles kaput) heterodimers, which are key regulators of biological rhythm. It is known as a transcript that regulates biological rhythm, but there has not been any report on the treatment of osteoarthritis through inhibition of DEC1 expression or activity.
상기 유전자의 뉴클레오티드 서열을 서열번호 1에 나타내었다. 또한, 상기 유전자는 뉴클레오티드 서열이 유전자 은행에 등록되어 있으므로 당업자라면 쉽게 입수가 가능할 것이다.The nucleotide sequence of the gene is shown in SEQ ID NO: 1. In addition, since the nucleotide sequence of the gene is registered in the gene bank, it will be readily available to those skilled in the art.
상기 DEC1의 발현 억제제는 DEC1에 특이적으로 결합하는 안티센스 올리고뉴클레오티드, siRNA, shRNA, microRNA 및 리보자임으로 이루어진 군으로부터 선택된 1종 이상을 포함할 수 있으나, 유전자의 발현을 억제시킬 수 있는 제제라면 제한 없이 포함될 수 있다.The DEC1 expression inhibitor may include one or more selected from the group consisting of antisense oligonucleotides, siRNA, shRNA, microRNA, and ribozymes that specifically bind to DEC1, but limited as long as it is an agent capable of inhibiting gene expression. Can be included without.
또한, 상기 DEC1 활성 억제제는 DEC1에 특이적으로 결합하는 화합물, 펩티드, 단백질, 앱타머 및 항체로 이루어진 군으로부터 선택된 1종 이상을 포함할 수 있으나, 유전자의 활성을 억제시킬 수 있는 제제라면 제한 없이 포함될 수 있다.In addition, the DEC1 activity inhibitor may include one or more selected from the group consisting of compounds, peptides, proteins, aptamers, and antibodies that specifically bind to DEC1, but any agent capable of inhibiting gene activity is not limited. Can be included.
본 발명에 따른 약학적 조성물은 유효성분 이외에 약제학적으로 허용되는 담체를 포함할 수 있다. 이때, 약제학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세 결정성셀룰로스, 폴리비닐피로리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필 히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 또한, 상기성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.The pharmaceutical composition according to the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient. At this time, the pharmaceutically acceptable carrier is commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , Polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, but are not limited thereto. In addition, a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like may be additionally included in addition to the above components.
본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구투여(예를 들어, 정맥 내, 피하, 복강 내 또는 국소에 적용)할 수 있으며, 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 시간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다.The pharmaceutical composition of the present invention can be administered orally or parenterally (for example, intravenous, subcutaneous, intraperitoneal or topical application) according to a desired method, and the dosage is Although it depends on the degree, drug form, administration route and time, it may be appropriately selected by those skilled in the art.
본 발명의 약학적 조성물은 약학적으로 유효한 양으로 투여한다. 본 발명에 있어서 "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효량은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "a pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective amount is the type, severity, activity of the drug, and It can be determined according to sensitivity, administration time, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field.
본 발명에 따른 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고, 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.
구체적으로 본 발명의 약학적 조성물의 유효량은 환자의 연령, 성별, 상태, 체중, 체내에 활성 성분의 흡수도, 불활성율 및 배설속도, 질병종류, 병용되는 약물에 따라 달라질 수 있으며, 일반적으로는 체중 1kg 당 0.001 내지 150mg, 바람직하게는 0.01 내지 100mg을 매일 또는 격일 투여하거나, 1일 1 내지 3회로 나누어 투여할 수 있다. 그러나 투여 경로, 비만의 중증도, 성별, 체중, 연령 등에 따라서 증감 될 수 있으므로 상기 투여량이 어떠한 방법으로도 본 발명의 범위를 한정하는 것은 아니다.Specifically, the effective amount of the pharmaceutical composition of the present invention may vary depending on the patient's age, sex, condition, weight, absorption of the active ingredient in the body, inactivation rate and excretion rate, the type of disease, and the drug to be used in combination. 0.001 to 150 mg, preferably 0.01 to 100 mg per 1 kg of body weight may be administered daily or every other day, or divided into 1 to 3 times a day. However, since it may increase or decrease depending on the route of administration, the severity of obesity, sex, weight, age, etc., the dosage amount does not limit the scope of the present invention in any way.
본 발명의 다른 양태는 상기 약학적 조성물을 개체에 투여하는 단계를 포함하는 골관절염 치료방법에 관한 것이다.Another aspect of the present invention relates to a method for treating osteoarthritis comprising administering the pharmaceutical composition to an individual.
상기 "개체"란 질병의 치료를 필요로 하는 대상을 의미하고, 보다 구체적으로는, 인간 또는 비-인간인 영장류, 마우스(mouse), 개, 고양이, 말 및 소 등의 포유류를 의미한다.The "individual" refers to a subject in need of treatment of a disease, and more specifically, refers to a mammal such as a human or non-human primate, mouse, dog, cat, horse, and cow.
본 발명의 또 다른 양태는 DEC1(Differentiated embryo chondrocyte expressed gene 1)의 발현 또는 활성 수준을 측정하는 제제를 포함하는 골관절염 진단용 조성물에 관한 것이다.Another aspect of the present invention relates to a composition for diagnosing osteoarthritis, including an agent for measuring the expression or activity level of DEC1 (Differentiated embryo chondrocyte expressed gene 1).
본 발명에서 사용되는 용어, "진단"이란 본 발명에 따른 조성물의 투여에 의해 병리 상태, 즉 골관절염의 존재 또는 특징을 확인하는 행위를 의미한다.The term "diagnosis" as used in the present invention means an act of confirming the presence or characteristics of a pathological condition, that is, osteoarthritis by administration of the composition according to the present invention.
본 발명에서, DEC1의 발현 또는 활성 수준을 측정하는 제제는 DEC1에 특이적으로 결합하는 프라이머, 프로브, 단백질 및 항체로 이루어진 군으로부터 선택된 1종 이상을 포함할 수 있으나, 유전자의 발현 또는 활성 수준을 측정할 수 있는 제제라면 제한 없이 포함될 수 있다.In the present invention, the agent for measuring the expression or activity level of DEC1 may include one or more selected from the group consisting of primers, probes, proteins and antibodies that specifically bind to DEC1, but the expression or activity level of the gene Any formulation that can be measured may be included without limitation.
본 발명의 또 다른 양태는 하기 단계를 포함하는 골관절염 진단을 위한 정보제공방법에 관한 것이다.Another aspect of the present invention relates to a method for providing information for diagnosing osteoarthritis comprising the following steps.
생물학적 시료에 존재하는 DEC1(Differentiated embryo chondrocyte expressed gene 1)의 발현 또는 활성 정도를 측정하는 단계; 및 Measuring the level of expression or activity of DEC1 (Differentiated embryo chondrocyte expressed gene 1) present in the biological sample; And
DEC1의 발현 또는 활성 정도를 정상 대조군의 발현 또는 활성 정도와 비교하는 단계.Comparing the level of expression or activity of DEC1 with the level of expression or activity of a normal control.
상기 DEC1의 발현 또는 활성 정도가 정상 대조군의 발현 또는 활성 정도와 비교하여 증가하는 경우 이를 골관절염으로 판정할 수 있다.When the level of expression or activity of DEC1 increases compared to the level of expression or activity of the normal control, it can be determined as osteoarthritis.
상기 방법은 포유류, 특히 인간을 대상으로 포함한다.The method includes targeting mammals, particularly humans.
구체적으로는, 상기 인간은 골관절염이 발병했을 것으로 의심되는 사람 또는 의심되지 않는 사람으로 골관절염 진단여부가 필요한 사람을 포함한다.Specifically, the human is a person suspected of developing osteoarthritis or a person who is not suspicious and includes a person who needs to be diagnosed with osteoarthritis.
상기 "생물학적 시료"는 조직, 세포, 혈액, 혈청, 혈장, 타액 및 뇨로 이루어질 수 있으나, 이에 제한되는 것은 아니다.The "biological sample" may be composed of tissue, cells, blood, serum, plasma, saliva, and urine, but is not limited thereto.
상기 시료는 검출에 사용하기 전에 전처리할 수 있으며, 예를 들어, 여과, 증류, 추출, 농축, 방해 성분의 불활성화, 시약의 첨가 등을 포함할 수 있다. 또한, 상기 시료로부터 핵산 및 단백질을 분리하여 검출에 사용할 수 있다.The sample may be pretreated before use for detection, and may include, for example, filtration, distillation, extraction, concentration, inactivation of interfering components, addition of reagents, and the like. In addition, nucleic acids and proteins can be separated from the sample and used for detection.
상기 DEC1의 발현 또는 활성 정도를 측정하는 단계는 중합효소반응(PCR), 역전사 중합효소반응(RT-PCR), 경쟁적 역전사 중합효소반응(Competitive RT-PCR), 실시간 역전사 중합효소반응(Realtime RT-PCR), RNase 보호 분석법(RPA; RNase protection assay), 노던 블롯팅(Northern blotting), DNA 칩, 웨스턴 블롯팅(Western blotting), ELISA(enzyme linked immunosorbent assay), 방사선 면역 분석(Radioimmunoassay, RIA), 방사 면역 확산법(radioimmunodiffusion), 오우크테로니(Ouchterlony) 면역확산법, 로케트(rocket) 면역전기영동, 면역침전 분석법(Immunoprecipitation Assay), 보체 고정 분석법(Complement Fixation Assay), 단백질 칩, 및 In vitro kinase assay(IKKB)로 이루어진 군으로부터 선택된 1종 이상의 방법으로 측정할 수 있으나, 이에 제한되는 것은 아니다.The step of measuring the level of expression or activity of DEC1 is a polymerase reaction (PCR), a reverse transcription polymerase reaction (RT-PCR), a competitive reverse transcription polymerase reaction (Competitive RT-PCR), and a realtime reverse transcription polymerase reaction (Realtime RT- PCR), RNase protection assay (RPA), Northern blotting, DNA chip, Western blotting, ELISA (enzyme linked immunosorbent assay), radioimmunoassay (RIA), Radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, Immunoprecipitation Assay, Complement Fixation Assay, protein chip, and in vitro kinase assay It can be measured by one or more methods selected from the group consisting of (IKKB), but is not limited thereto.
본 발명의 또 다른 양태는 개체 내 DEC1(Differentiated embryo chondrocyte expressed gene 1)의 발현 또는 활성 정도를 감소시키는 물질을 선별하는 단계를 포함하는 골관절염 치료제의 스크리닝 방법에 관한 것이다.Another aspect of the present invention relates to a screening method for a therapeutic agent for osteoarthritis comprising the step of selecting a substance that decreases the expression or activity of DEC1 (Differentiated embryo chondrocyte expressed gene 1) in an individual.
상기 방법에서 개체 내 DEC1의 발현 또는 활성 정도를 측정하고 골관절염 치료용 후보물질 투여 시, 상기 DEC1의 발현 또는 활성 정도가 기존 대비 감소되는 경우 상기 후보물질은 골관절염 치료제로 이용될 수 있다.In the above method, when the degree of expression or activity of DEC1 in an individual is measured and a candidate substance for osteoarthritis treatment is administered, the candidate substance may be used as a treatment for osteoarthritis when the expression or activity degree of the DEC1 decreases compared to the existing one.
상기 방법은 개체에 대상 물질 처리 전에 개체 내 DEC1의 발현 또는 활성 정도를 측정하고 처리 후에 이를 측정하여 비교할 수도 있고, 여러 개체 중 일부 개체에 대상 물질을 처리하여 비교군과 상기 발현 또는 활성 정도를 비교할 수도 있다.The above method may measure the expression or activity level of DEC1 in the subject before treatment with the subject substance to the subject, and measure and compare it after treatment, or compare the expression or activity level with the control group by treating the subject substance in some of several subjects. May be.
상기 DEC1의 발현 또는 활성 정도를 측정하는 단계는 중합효소반응(PCR), 역전사 중합효소반응(RT-PCR), 경쟁적 역전사 중합효소반응(Competitive RT-PCR), 실시간 역전사 중합효소반응(Realtime RT-PCR), RNase 보호 분석법(RPA; RNase protection assay), 노던 블롯팅(Northern blotting), DNA 칩, 웨스턴 블롯팅(Western blotting), ELISA(enzyme linked immunosorbent assay), 방사선 면역 분석(Radioimmunoassay, RIA), 방사 면역 확산법(radioimmunodiffusion), 오우크테로니(Ouchterlony) 면역확산법, 로케트(rocket) 면역전기영동, 면역침전 분석법(Immunoprecipitation Assay), 보체 고정 분석법(Complement Fixation Assay), 단백질 칩, 및 In vitro kinase assay(IKKB)로 이루어진 군으로부터 선택된 1종 이상의 방법으로 측정할 수 있으나, 이에 제한되는 것은 아니다.The step of measuring the level of expression or activity of DEC1 is a polymerase reaction (PCR), a reverse transcription polymerase reaction (RT-PCR), a competitive reverse transcription polymerase reaction (Competitive RT-PCR), and a realtime reverse transcription polymerase reaction (Realtime RT- PCR), RNase protection assay (RPA), Northern blotting, DNA chip, Western blotting, ELISA (enzyme linked immunosorbent assay), radioimmunoassay (RIA), Radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, Immunoprecipitation Assay, Complement Fixation Assay, protein chip, and in vitro kinase assay It can be measured by one or more methods selected from the group consisting of (IKKB), but is not limited thereto.
상기 방법에서 개체에 대상 물질을 처리하는 방법은 특별히 한정되지 않으며, 예를 들면 개체로부터 얻어진 생물학적 시료에 처리할 수 있다. 생물학적 시료는 전술한 바의 시료일 수 있다.In the above method, a method of treating a target substance on an individual is not particularly limited, and for example, a biological sample obtained from an individual can be treated. The biological sample may be a sample as described above.
상기 방법에서 "대상 물질"은 본 발명의 DEC1의 발현 또는 활성 정도에 영향을 미치는지 여부를 검사하기 위하여 스크리닝에서 이용되는 미지의 물질을 의미한다. 상기 대상 물질은 화학물질, 뉴클레오타이드, 안티센스-RNA, siRNA(small interference RNA) 및 천연물 추출물을 포함하나, 이에 한정되는 것은 아니다.In the above method, "target substance" means an unknown substance used in screening to test whether it affects the expression or activity level of DEC1 of the present invention. The target substance includes, but is not limited to, a chemical substance, a nucleotide, an antisense-RNA, a small interference RNA (siRNA), and a natural product extract.
본 발명은 골관절염 예방 또는 치료용 조성물, 진단용 조성물, 진단을 위한 정보제공방법 및 치료제의 스크리닝 방법에 관한 것이다. 본 발명으로 골관절염의 진단이 가능할 것으로 기대되며, 본 발명의 마커인 DEC1의 발현 및 활성을 조절하여 골관절염 치료제 개발에 유용하게 사용할 수 있을 것으로 기대된다. The present invention relates to a composition for preventing or treating osteoarthritis, a composition for diagnosis, a method for providing information for diagnosis, and a method for screening a therapeutic agent. The present invention is expected to enable the diagnosis of osteoarthritis, and is expected to be useful in the development of osteoarthritis therapeutic agents by controlling the expression and activity of DEC1, a marker of the present invention.
도 1a 및 1b는 본 발명의 일 실시예에 따라 DEC1 녹아웃(knockout) 마우스의 연골조직에서의 연골손상 정도 및 DEC1 발현을 확인한 도이다.
도 2a 및 2b는 본 발명의 일 실시예에 따라 IL-1b에 의한 염증 유도에 따른 연골세포에서의 DEC1 발현을 확인한 도이다.
도 2c 및 2d는 본 발명의 일 실시예에 따라 TNF-α에 의한 염증 유도에 따른 연골세포에서의 DEC1 발현을 확인한 도이다.
도 3a 및 3b는 본 발명의 일 실시예에 따라 연골조직 및 연골세포에서 DEC1 과발현에 따른 효과를 확인한 도이다.
도 3c 및 3d는 본 발명의 일 실시예에 따라 연골조직 및 연골세포에서 DEC1 발현 억제에 따른 효과를 확인한 도이다.
도 4a 및 4b는 본 발명의 일 실시예에 따라 DEC1 과발현에 따른 연골조직에서의 연골손상 정도를 확인한 도이다.1A and 1B are diagrams illustrating the degree of cartilage damage and DEC1 expression in cartilage tissue of a DEC1 knockout mouse according to an embodiment of the present invention.
2A and 2B are diagrams illustrating the expression of DEC1 in chondrocytes according to inflammation induction by IL-1b according to an embodiment of the present invention.
2C and 2D are diagrams confirming the expression of DEC1 in chondrocytes according to inflammation induction by TNF-α according to an embodiment of the present invention.
3A and 3B are diagrams illustrating the effect of DEC1 overexpression in cartilage tissue and cartilage cells according to an embodiment of the present invention.
3C and 3D are diagrams illustrating the effect of inhibiting DEC1 expression in cartilage tissue and chondrocytes according to an embodiment of the present invention.
4A and 4B are diagrams illustrating the degree of cartilage damage in cartilage tissue according to DEC1 overexpression according to an embodiment of the present invention.
이하, 실시예를 통하여 본 발명을 더욱 상세히 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 through examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .
준비예. 샘플 준비Preparation Yes. sample preparation
DEC1 녹아웃(knockout) 마우스 및 야생형 마우스는 JACKSON LAB(미국)에서 구입하였다.DEC1 knockout mice and wild-type mice were purchased from JACKSON LAB (USA).
실험예 1. DEC1 녹아웃(knockout) 마우스의 연골조직에서의 DEC1 발현 확인Experimental Example 1. DEC1 expression confirmation in cartilage tissue of DEC1 knockout mouse
9주령 마우스(DEC1 녹아웃 마우스 및 야생형 마우스)의 무릎 관절의 아래 쪽 연골인 반월판 파괴를 유도시켜(destabilization of medial meniscus; DMM) 관절염(연골 손상)을 유도하였다. 8주 경과 후, 관절조직을 채취하여 하기와 같이 실험하였다. 이때, 각 DEC1 녹아웃 마우스 및 야생형 마우스 그룹에서 DMM을 수행하지 않은 마우스를 대조군(Sham 마우스)으로 사용하였다.Arthritis (cartilage damage) was induced by inducing meniscus destruction (destabilization of medial meniscus; DMM) in the lower cartilage of the knee joint of 9-week-old mice (DEC1 knockout mice and wild-type mice). After 8 weeks, the joint tissue was collected and tested as follows. At this time, mice that did not perform DMM in each DEC1 knockout mouse and wild-type mouse group were used as a control (Sham mouse).
먼저, 관절조직을 4% 파라포름알데하이드로 고정시킨 후, 0.5 M EDTA 용액으로 3일 간 탈회(decalcification) 과정을 진행하였다. 그 다음, 조직을 30%, 50%, 70%, 90%, 95% 및 100% 에탄올에 담가 탈수 과정을 진행한 후, 자일렌 및 파라핀을 침투시켰다. 조직을 5 um 두께로 잘라 슬라이드 위에 올려 보관하였다.First, after fixing the joint tissue with 4% paraformaldehyde, a decalcification process was performed for 3 days with 0.5 M EDTA solution. Then, the tissue was immersed in 30%, 50%, 70%, 90%, 95%, and 100% ethanol to undergo a dehydration process, and then xylene and paraffin were infiltrated. The tissue was cut to a thickness of 5 um and stored on a slide.
자일렌, 각 100%, 95%, 90%, 70%, 50% 및 30% 에탄올, 및 물의 순서로 재수화(rehydration) 과정을 거친 조직 절편을 페마톡실린(hematoxylin), 0.1% 패스트그린(fast green) 및 0.1% 사프라닌 O(safranin O) 용액으로 염색하고 70%, 90%, 95% 및 100% 에탄올, 및 자일렌에 담근 후, 고정액을 뿌려 고정하였다. 현미경으로 염색된 정도를 확인하여 연골조직의 손상 정도를 OARSI(Osteoarthritis Research Society International grade) 가이드라인에 따라 점수화하였다.Xylene, each 100%, 95%, 90%, 70%, 50% and 30% ethanol, and water in the order of rehydration (hematoxylin), 0.1% fast green ( fast green) and 0.1% safranin O solution and immersed in 70%, 90%, 95% and 100% ethanol, and xylene, and then sprinkled with a fixative to fix. The degree of staining was checked under a microscope, and the degree of damage to the cartilage tissue was scored according to OARSI (Osteoarthritis Research Society International grade) guidelines.
다음으로, 상기 재수화 과정을 거친 조직 절편을 3% H2O2 용액에 5분 간 담가 조직 내 퍼옥시다아제(peroxidase) 활성을 억제하고, 0.1 % 트립신 및 0.1% 트리톤 X-100 용액에 각각 5분 간 담가 조직 내 항원을 노출시켰다. 이후 1% 소 혈청 알부민(bovine serum albumin; BSA) 용액에 1시간 동안 담가 블로킹 시키고, DEC1 일차 항체(Sigma aldrich)에 2시간 동안 반응시켰다. 세척 후, 이차 항체-스트렙트아비딘-HRP 용액(Dako EnVision HRP)에 30분 간 반응시키고, AEC substrate chromogen 용액(DAKO)에 10분간 반응시켜 발색을 진행하였다. 세척 후 고정하여 현미경으로 DEC1의 발현을 확인하였다.Next, the tissue section that has undergone the rehydration process is immersed in 3% H 2 O 2 solution for 5 minutes to inhibit peroxidase activity in the tissue, and 5 in 0.1% trypsin and 0.1% Triton X-100 solution, respectively. Soak for minutes to expose the antigen in the tissue. Thereafter, it was immersed in 1% bovine serum albumin (BSA) solution for 1 hour to block, and reacted with DEC1 primary antibody (Sigma aldrich) for 2 hours. After washing, it was reacted with a secondary antibody-streptavidin-HRP solution (Dako EnVision HRP) for 30 minutes, and reacted with an AEC substrate chromogen solution (DAKO) for 10 minutes to develop color. After washing and fixing, the expression of DEC1 was confirmed under a microscope.
도 1a 및 1b에서 확인할 수 있듯이, 야생형(Wild Type) Sham 마우스와 DMM을 수행한 마우스의 연골 손상 정도 차이에 비하여, DEC1 녹아웃(DEC1 KO) Sham 마우스와 DMM을 수행한 마우스의 연골 손상 정도 차이가 보다 감소함을 알 수 있었다. 또한, DEC1 녹아웃 마우스에 비하여, 야생형 마우스의 연골 손상을 유도한 연골조직에서 DEC1 단백질의 발현이 증가함을 알 수 있었다.As can be seen in Figures 1a and 1b, compared to the difference in cartilage damage between wild type Sham mice and mice subjected to DMM, the difference in cartilage damage between DEC1 knockout (DEC1 KO) Sham mice and mice subjected to DMM is It can be seen that it is reduced more. In addition, compared to DEC1 knockout mice, it was found that the expression of the DEC1 protein was increased in cartilage tissue that induced cartilage damage in wild-type mice.
실험예 2. 염증 유도에 따른 연골세포에서의 DEC1 발현 확인Experimental Example 2. Confirmation of DEC1 expression in chondrocytes according to inflammation induction
5일령 마우스의 무릎 조직을 채취하고, 0.2% 제2형 콜라게나아제(type II collagenase)(Sigma Aldrich) 및 0.02% 트립신을 3시간 동안 처리하여 무릎 조직의 피부 및 근육을 제거하였다. 그 다음, 연골조직만을 분리하여 0.2% 제2형 콜라게나아제로 조직을 녹여 연골세포를 확보한 후, DMEM 배양액을 사용하여 3x105 개/1 dish의 농도로 연골세포를 배양하였다. 배양 이틀째에 DMEM 배양액을 교체하고, 각 IL-1b 또는 TNF-α를 24시간 처리 후 세포를 수거하였다.The knee tissue of a 5-day-old mouse was collected and treated with 0.2% type II collagenase (Sigma Aldrich) and 0.02% trypsin for 3 hours to remove skin and muscles of the knee tissue. Then, only the cartilage tissue was separated and the tissue was melted with 0.2
상기 연골세포에 트리졸 용액을 첨가하고 클로로포름 용액을 첨가 후 원심분리(13,000rpm, 15분)하여 조직 내 단백질을 분리하였다. mRNA가 녹아 있는 수용액에 아이소프로판올을 첨가 후 원심분리(13,000rpm, 15분)하여 수분을 제거하였다. 수분이 제거된 침전물에 75% 에탄올을 첨가 후 원심분리(13,000rpm, 10분)하여 남아 있는 유기용매를 녹여 제거한 후, 공기 중에서 건조시켜 에탄올을 증발시켰다. 확보한 RNA에 역전사효소(reverse transcriptase)를 첨가하여 cDNA를 수득하고, cDNA에 SYBR, DEC1 및 생체주기(circadian clock) 유전자들의 프라이머(BMAL1; Forward 5'-AATCGCAAGAGGAAAGGCAGTG-3', Revers 5'-TGCTTCTGTGTATGGGTTGGTG-3', CLOCK; Forward 5'-GGCTGAAAGACGGCGAGAAC-3', Revers 5'-AAATGCTGTCTGGGAGGAGTGC-3', PER2; Forward 5'-TGGAGCAGGTTGAGGGCATTAC-3', Revers 5'-TCTCATTCTCGTGGTGTTTCCC-3')를 각각 첨가 후 qRT-PCR(정량적 real time-polymerase chain reaction)을 진행하여 DEC1을 포함한 생체주기 유전자들의 발현 정도를 정량화하였다.Trizol solution was added to the chondrocytes, chloroform solution was added, and then centrifuged (13,000 rpm, 15 minutes) to separate proteins in the tissue. Isopropanol was added to the aqueous solution in which the mRNA was dissolved, followed by centrifugation (13,000 rpm, 15 minutes) to remove moisture. 75% ethanol was added to the moisture-removed precipitate, centrifuged (13,000 rpm, 10 minutes) to dissolve and remove the remaining organic solvent, and dried in air to evaporate ethanol. CDNA was obtained by adding reverse transcriptase to the obtained RNA, and primers of SYBR, DEC1 and circadian clock genes (BMAL1; Forward 5'-AATCGCAAGAGGAAAGGCAGTG-3', Revers 5'-TGCTTCTGTGTATGGGTTGGTG) to cDNA -3', CLOCK; Forward 5'-GGCTGAAAGACGGCGAGAAC-3', Revers 5'-AAATGCTGTCTGGGAGGAGTGC-3', PER2; Forward 5'-TGGAGCAGGTTGAGGGCATTAC-3', Revers 5'-TCTCATTCTCGTGGTGTTTCCC-3') after each addition and qRT- PCR (quantitative real time-polymerase chain reaction) was performed to quantify the level of expression of life cycle genes including DEC1.
도 2a 내지 2d에서 확인할 수 있듯이, 사이토카인(IL-1b 및 TNF-α)을 처리한 연골세포에서 DEC1 mRNA의 발현이 증가함을 알 수 있었다.As can be seen in Figures 2a to 2d, it was found that the expression of DEC1 mRNA was increased in chondrocytes treated with cytokines (IL-1b and TNF-α).
실험예 3. 연골조직 및 연골세포에서 DEC1 발현 조절에 따른 효과 확인Experimental Example 3. Checking the effect of controlling DEC1 expression in cartilage tissue and cartilage cells
5일령 마우스의 무릎 조직을 채취하고, 0.2% 제2형 콜라게나아제 및 0.02% 트립신을 3시간 동안 처리하여 무릎 조직의 피부 및 근육을 제거하였다. 그 다음, 연골조직만을 분리하여 0.2% 제2형 콜라게나아제로 조직을 녹여 연골세포를 확보한 후, DMEM 배양액을 사용하여 3x105 개/1 dish의 농도로 연골세포를 배양하였다.The knee tissue of a 5-day-old mouse was collected and treated with 0.2
먼저, 배양 3일째에 DEC1 아데노바이러스를 2시간 동안 감염시키고, 24 시간 후 연골세포로부터 cDNA를 확보하였다. 그 다음, DEC1, Mmp3, Mmp13, Ptgs2, Il6 및 Adamts4의 프라이머(Mmp3; Forward 5'-CTGTGTGTGGTTGTGTGCTCATCCTAC-3', Revers 5'-GGCAAATCCGGTGTATAATCCACAATC-3', Mmp13; Forward 5'-TGATGGACCTTCTGGTCTTCTGGC-3', Revers 5'-CATCCACATGGTTGGGAAGTTCTG-3', Ptgs2; Forward 5'-CCAAACCAGCAGACTCATACTCATAG-3', Revers 5'-CATCTCTCTGCTCTGGTCAATGGAG-3', Il6; Forward 5'-AGAGATACAAAGAAATGATGGATGC-3', Revers 5'-CTAGGTTTGCCGAGTAGATCTC-3', Adamts4; Forward 5'-CATCCGAAACCCTGTCAA-3', Revers 5'-GCCCATCATCTTCCACAA-3')를 이용하여 qRT-PCR을 진행하여 해당 유전자를 증폭시켰다.First, on the third day of culture, DEC1 adenovirus was infected for 2 hours, and cDNA was obtained from chondrocytes after 24 hours. Then, the primers of DEC1, Mmp3, Mmp13, Ptgs2, Il6 and Adamts4 (Mmp3; Forward 5'-CTGTGTGTGGTTGTGTGCTCATCCTAC-3', Revers 5'-GGCAAATCCGGTGTATAATCCACAATC-3', Mmp13; Forward 5'-TGATGGACCTTC', Revers 5'-TGATGGACCTTC', Revers 5 '-CATCCACATGGTTGGGAAGTTCTG-3', Ptgs2; Forward 5'-CCAAACCAGCAGACTCATACTCATAG-3', Revers 5'-CATCTCTCTGCTCTGGTCAATGGAG-3', Il6; Forward 5'-AGAGATACAAATCGAAATGATGGATGC-3', Revers 5'-CTAGAGGTTTGCCGAG', Adamts4; 5'-CATCCGAAACCCTGTCAA-3', Revers 5'-GCCCATCATCTTCCACAA-3') was used to perform qRT-PCR to amplify the corresponding gene.
도 3a 및 3b에서 확인할 수 있듯이, 연골세포에서 DEC1의 과발현에 따라 연골기질 분해효소인 Mmp3, Mmp13 및 Adamts4, 및 염증 매개인자인 IL6 및 Ptgs2의 발현이 증가함을 알 수 있었다.As can be seen in Figures 3a and 3b, it was found that the expression of cartilage matrix degrading enzymes Mmp3, Mmp13 and Adamts4, and inflammatory mediators IL6 and Ptgs2 increased according to the overexpression of DEC1 in chondrocytes.
다음으로, 상기 배양된 연골세포에 DEC1 siRNA를 48시간동안 형질전환시켜 cDNA를 확보하였다. 그 다음, DEC1, Mmp3, Mmp13, Ptgs2, Il6 및 Adamts4의 프라이머(Mmp3; Forward 5'-CTGTGTGTGGTTGTGTGCTCATCCTAC-3', Revers 5'-GGCAAATCCGGTGTATAATCCACAATC-3', Mmp13; Forward 5'-TGATGGACCTTCTGGTCTTCTGGC-3', Revers 5'-CATCCACATGGTTGGGAAGTTCTG-3', Ptgs2; Forward 5'-CCAAACCAGCAGACTCATACTCATAG-3', Revers 5'-CATCTCTCTGCTCTGGTCAATGGAG-3', Il6; Forward 5'-AGAGATACAAAGAAATGATGGATGC-3', Revers 5'-CTAGGTTTGCCGAGTAGATCTC-3', Adamts4; Forward 5'-CATCCGAAACCCTGTCAA-3', Revers 5'-GCCCATCATCTTCCACAA-3')를 이용하여 qRT-PCR을 진행하여 해당 유전자를 증폭시켰다.Next, the cultured chondrocytes were transformed with DEC1 siRNA for 48 hours to obtain cDNA. Then, primers of DEC1, Mmp3, Mmp13, Ptgs2, Il6 and Adamts4 (Mmp3; Forward 5'-CTGTGTGTGGTTGTGTGCTCATCCTAC-3', Revers 5'-GGCAAATCCGGTGTATAATCCACAATC-3', Mmp13; Forward 5'-TGATGGACCTTC', Revers 5'-TGATGGACCTTC', Revers 5 '-CATCCACATGGTTGGGAAGTTCTG-3', Ptgs2; Forward 5'-CCAAACCAGCAGACTCATACTCATAG-3', Revers 5'-CATCTCTCTGCTCTGGTCAATGGAG-3', Il6; Forward 5'-AGAGATACAAATCGAAATGATGGATGC-3', Revers 5'-CTAGAGGTTTGCCGAG', Adamts4; 5'-CATCCGAAACCCTGTCAA-3', Revers 5'-GCCCATCATCTTCCACAA-3') was used to perform qRT-PCR to amplify the corresponding gene.
도 3c 및 3d에서 확인할 수 있듯이, 연골세포에서 DEC1의 발현 억제에 따라 연골기질 분해효소인 Mmp3, Mmp13, Adamts4, 및 염증 매개인자인 Il6의 발현이 감소함을 알 수 있었다.As can be seen in Figures 3c and 3d, it was found that the expression of the cartilage matrix degrading enzymes Mmp3, Mmp13, Adamts4, and the inflammatory mediator Il6 decreased according to the inhibition of DEC1 expression in chondrocytes.
추가적으로, 8주령 마우스의 무릎에 1x109 PFU 분량(10ul)의 DEC1 아데노바이러스를 1주에 한번씩 총 3번 주입하고, 4주 후 무릎 조직을 확보하였다. 그 다음, 상기 실험예 2의 방법으로 사프라닌 O 염색을 진행하여 연골조직의 손상 정도를 OARSI(Osteoarthritis Research Society International grade) 가이드라인에 따라 점수화하였다.In addition, a 1× 10 9 PFU amount (10ul) of DEC1 adenovirus was injected into the knee of an 8-week-old mouse once a week for a total of 3 times, and knee tissue was secured after 4 weeks. Then, safranin O staining was performed by the method of Experimental Example 2, and the degree of damage to the cartilage tissue was scored according to OARSI (Osteoarthritis Research Society International grade) guidelines.
도 4a 및 4b에서 확인할 수 있듯이, DEC1의 과발현에 의해 연골조직의 손상이 증가하여 관절염이 발병함을 알 수 있었다.As can be seen in Figures 4a and 4b, it was found that the damage to the cartilage tissue increased due to the overexpression of DEC1, resulting in arthritis.
<110> INDUSTRY FOUNDATION OF CHONNAM NATIONAL UNIVERSITY
<120> Composition for preventing and treating arthritis comprising a
DEC1 expression and activity modulator
<130> PN190012
<160> 1
<170> KoPatentIn 3.0
<210> 1
<211> 1236
<212> DNA
<213> Unknown
<220>
<223> Differentiated embryo chondrocyte expressed gene 1
<400> 1
atggaacgga tccccagcgc gcaaccacct cctacctgcc tgcccaaagc tccagggctg 60
gagcacggag acctgtcagg gatggatttt gcccacatgt accaagtgta caagtccagg 120
cggggaataa aacggagcga agacagcaag gaaacttaca aactgccgca ccggctgatt 180
gagaaaaaga gacgtgaccg gattaacgag tgcattgccc agctgaagga tctcctaccc 240
gaacatctca aacttactac tttgggtcac ttggaaaaag cagtggttct ggagcttacg 300
ttgaagcacg tgaaagcatt gacaaatcta attgatcagc agcagcagaa aatcattgcc 360
ctgcagagcg gtttacaagc tggtgatttg tcgggaagaa atctcgaggc agggcaagaa 420
atgttctgct caggtttcca gacttgtgcc cgtgaggtac ttcagtacct ggcgaagcat 480
gagaacactc gggacctgaa atcttcccag ctcgtcactc atctccatcg tgtggtctcg 540
gagctgctgc agggtggtgc ttccaggaaa ccattggact cggctcccaa agccgtggac 600
ttgaaagaga agcccagctt cctagccaag ggatcagaag gccctgggaa aaactgtgtg 660
ccagtcatcc agcggacttt tgctccctcg ggtggggagc agagcggcag tgacacggac 720
acagacagtg gctatggagg tgaattggag aagggggact tgcgcagtga gcagccgtac 780
ttcaaaagcg accatggacg caggttcgcc gtgggagaac gtgtcagcac aattaagcaa 840
gaatccgaag agccccccac caaaaagagc cgaatgcagc tctcagaaga ggaaggccac 900
ttcgcgggca gtgatctgat gggttcccca tttcttgggc cacacccaca tcagcctcct 960
ttttgccttc ccttctatct catcccacca tcggccactg cctacctgcc tatgctggag 1020
aaatgctggt accccacctc tgtgccagtg ttatacccag gcctcaacac ctcagctgca 1080
gccctctcca gcttcatgaa cccagacaag ataccgactc ccttgcttct gccccagaga 1140
ctcccttctc ctttggcaca ttcgtccctc gactcttcgg ccttgctcca ggctttgaag 1200
cagatccctc ctttaaactt agaaaccaaa gactaa 1236
<110> INDUSTRY FOUNDATION OF CHONNAM NATIONAL UNIVERSITY
<120> Composition for preventing and treating arthritis comprising a
DEC1 expression and activity modulator
<130> PN190012
<160> 1
<170> KoPatentIn 3.0
<210> 1
<211> 1236
<212> DNA
<213> Unknown
<220>
<223> Differentiated embryo chondrocyte expressed
Claims (8)
생물학적 시료에 존재하는 DEC1(Differentiated embryo chondrocyte expressed gene 1)의 발현 또는 활성 정도를 측정하는 단계; 및
DEC1의 발현 또는 활성 정도를 정상 대조군의 발현 또는 활성 정도와 비교하는 단계.Information providing method for diagnosing osteoarthritis comprising the following steps:
Measuring the level of expression or activity of DEC1 (Differentiated embryo chondrocyte expressed gene 1) present in the biological sample; And
Comparing the level of expression or activity of DEC1 with the level of expression or activity of a normal control.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004035827A2 (en) * | 2002-10-18 | 2004-04-29 | Inserm | Microarrays allowing molecular profiling of rheumatoid arthritis comparatively to osteoarthritis andtheir use |
| KR20140118457A (en) | 2013-03-29 | 2014-10-08 | (주)생명의나무 | pharmaceutical composition for preventing or treating bone diseases comprising spinasterol derivative sugar compound |
| KR20180034291A (en) * | 2016-09-27 | 2018-04-04 | 가톨릭대학교 산학협력단 | Kit for treating osteoarthritis with alleviating pain |
| US20180369327A1 (en) * | 2017-06-26 | 2018-12-27 | Dong-A University Research Foundation For Industry-Academy Cooperation | Method for treating or preventing osteoarthritis |
| KR20190003413A (en) * | 2017-06-30 | 2019-01-09 | 코오롱생명과학 주식회사 | A Pharmaceutical Composition For Preventing or Treating Osteoarthritis |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004035827A2 (en) * | 2002-10-18 | 2004-04-29 | Inserm | Microarrays allowing molecular profiling of rheumatoid arthritis comparatively to osteoarthritis andtheir use |
| KR20140118457A (en) | 2013-03-29 | 2014-10-08 | (주)생명의나무 | pharmaceutical composition for preventing or treating bone diseases comprising spinasterol derivative sugar compound |
| KR20180034291A (en) * | 2016-09-27 | 2018-04-04 | 가톨릭대학교 산학협력단 | Kit for treating osteoarthritis with alleviating pain |
| US20180369327A1 (en) * | 2017-06-26 | 2018-12-27 | Dong-A University Research Foundation For Industry-Academy Cooperation | Method for treating or preventing osteoarthritis |
| KR20190003413A (en) * | 2017-06-30 | 2019-01-09 | 코오롱생명과학 주식회사 | A Pharmaceutical Composition For Preventing or Treating Osteoarthritis |
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| KR102275313B1 (en) | 2021-07-09 |
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