KR102655961B1 - Anti-aging composition comprising 4-hexylresorcinol as an active ingredient - Google Patents
Anti-aging composition comprising 4-hexylresorcinol as an active ingredient Download PDFInfo
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- KR102655961B1 KR102655961B1 KR1020210075500A KR20210075500A KR102655961B1 KR 102655961 B1 KR102655961 B1 KR 102655961B1 KR 1020210075500 A KR1020210075500 A KR 1020210075500A KR 20210075500 A KR20210075500 A KR 20210075500A KR 102655961 B1 KR102655961 B1 KR 102655961B1
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- aging
- present
- hexylresorcinol
- active ingredient
- composition
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Abstract
본 발명은 4-헥실레조르시놀(4-hexylresorcinol)을 유효성분으로 포함하는 항노화용 조성물에 관한 것으로, 4-헥실레조르시놀(4-hexylresorcinol)이 혈관내피세포에서 항노화와 관련된 Sirt6 유전자의 발현을 촉진시키는 것을 확인함으로써, 4-헥실레조르시놀(4-hexylresorcinol)을 항노화 제제로 제공한다.The present invention relates to an anti-aging composition containing 4-hexylresorcinol as an active ingredient, wherein 4-hexylresorcinol is an inhibitor of the Sirt6 gene related to anti-aging in vascular endothelial cells. By confirming that it promotes expression, 4-hexylresorcinol is provided as an anti-aging agent.
Description
본 발명은 4-헥실레조르시놀(4-hexylresorcinol)을 유효성분으로 포함하는 항노화용 조성물에 관한 것이다.The present invention relates to an anti-aging composition containing 4-hexylresorcinol as an active ingredient.
노화는 시간이 흐름에 따라 생물의 신체기능이 퇴화하는 현상이다. 세포의 노화는 세포가 분열할 수 있는 능력을 잃어버리는 것으로 나타난다. 줄기세포 역시 세포군이기 때문에 점차 그 기능을 서서히 소실하게 된다. 일반적으로 세포의 노화는 Rb(retinoblastoma) 단백질에 의존적인 노화기전과 염색체를 보호하는 역할을 담당하는 텔로미어(telomere)의 손실로 일어나는 노화, 항산화 스트레스에 의한 노화, DNA 손상에 의한 노화 등이 있다. 이러한 DNA 손상신호 전달기전과 스트레스 신호전달 기전이 외부 환경적 요인에 의해 발생할 수 있는 세포의 변형을 억제하는 종양억제 신호전달과 많은 부분을 공유하며 많은 관련 유전자들에 대한 연구가 진행되고 있다.Aging is a phenomenon in which an organism's physical functions deteriorate over time. Cellular aging occurs when cells lose their ability to divide. Since stem cells are also a group of cells, they gradually lose their function. In general, cellular aging includes aging mechanisms dependent on the Rb (retinoblastoma) protein, aging caused by loss of telomeres that protect chromosomes, aging caused by antioxidant stress, and aging caused by DNA damage. These DNA damage signal transduction mechanisms and stress signal transduction mechanisms share many aspects with tumor suppressor signal transduction, which suppresses cell deformation that may occur due to external environmental factors, and research on many related genes is in progress.
다양한 노화 관련 인자들중에서 Deacetylation 효소 활성을 갖는 Sirtuin family. Sirtuin family는 포유동물에서 Sirt1, Sirt2, Sirt3, Sirt4, Sirt5, Sirt6와 Sirt7로 이루어져 있으며, 수명연장 조절단백질로 간주되어 왔다.Among various aging-related factors, the Sirtuin family has deacetylation enzyme activity. The Sirtuin family consists of Sirt1, Sirt2, Sirt3, Sirt4, Sirt5, Sirt6, and Sirt7 in mammals, and has been considered a protein that regulates lifespan extension.
서투인1(Sirtuin1; SIRT1)은 만성염증, 암, 노화를 조절하는 유전자에 속하며, SIR2라는 이스트의 유전자가 에너지 처리과정을 조절하는 마스터 스위치 역할을 하는 것이라고 보고된 바 있다. SIR2 유전자가 활성화된 세포는 평균수명보다 오래 사는 반면, SIR2 유전자가 결여된 세포는 수명이 짧음이 증명되었다. 그 후 꼬마선충, 초파리의 경우 SIR2가 과잉발현되면 수명이 연장되는 것으로 밝혀졌으며, 포유류의 SIRT1 유전자가 SIR2와 등가이며, 동일한 작용을 한다는 연구결과가 발표된 바 있다. 인간의 SIRT1은 히스톤과 비히스톤 단백질을 탈아세틸화하여 염증매개물질을 조절하는 NAD+ 의존성 단백질/히스톤 탈아세틸화효소로써, 항노화ㆍ항염 단백질로 알려져 있다. 최근에는 SIRT1이 스트레스, 대사, 세포사멸, 조로 등에 긍정적 역할을 미친다는 연구결과들이 발표되고 있다.Sirtuin1 (SIRT1) belongs to a gene that regulates chronic inflammation, cancer, and aging, and it has been reported that a yeast gene called SIR2 acts as a master switch that regulates energy processing. It has been proven that cells with the SIR2 gene activated live longer than average, while cells lacking the SIR2 gene have a shorter lifespan. Afterwards, in the case of Caenorhabditis elegans and fruit flies, overexpression of SIR2 was found to extend the lifespan, and research results were published showing that the mammalian SIRT1 gene is equivalent to SIR2 and has the same function. Human SIRT1 is an NAD+-dependent protein/histone deacetylase that regulates inflammatory mediators by deacetylating histone and non-histone proteins, and is known as an anti-aging and anti-inflammatory protein. Recently, research results have been published showing that SIRT1 plays a positive role in stress, metabolism, apoptosis, and premature aging.
Sirtuin family 중에서도 Sirt1은 효모 Sir2 단백질과 가장 유사한 아미노산 배열을 갖고 있어서, 연구결과에 따르면 Sirt1 KO 마우스는 Embryonic lethal 하며, 조직 특이적으로 Sirt1을 결실 시 Sirt1이 노인성 질환과 (퇴행성신경질환, 당뇨, 암, 등) 과 밀접한 연관성이 있음을 보여주었으며, Sirt1를 과발현 하면 노인성 질환이 감소됨을 확인되었다. Sirt2, 3, 4, 5, 및 7 각각의 KO 마우스는 암발생 증가, Hearing loss, 심장질환 등 다양한 노인성 관련 질환 표현형들이 관찰되었으며, Sirt6 KO 마우스는 조로증(Premature aging)과 유사한 표현형을 보여주었고, Sirt6 과발현 transgenic 마우스는 수명이 약 15-20% 증가한다는 연구 결과가 발표되었다.Among the Sirtuin family, Sirt1 has an amino acid sequence most similar to the yeast Sir2 protein, and according to research results, Sirt1 KO mice are embryonic lethal, and when tissue-specific Sirt1 is deleted, Sirt1 is associated with geriatric diseases (degenerative neuropathy, diabetes, and cancer). , etc.), and it was confirmed that overexpressing Sirt1 reduces geriatric diseases. Sirt2, 3, 4, 5, and 7 KO mice showed various age-related disease phenotypes, such as increased cancer incidence, hearing loss, and heart disease, while Sirt6 KO mice showed a phenotype similar to premature aging. Research has shown that transgenic mice overexpressing Sirt6 have an increased lifespan of approximately 15-20%.
본 발명은 항노화용 조성물을 제공하기위해, 4-헥실레조르시놀(4-hexylresorcinol)이 혈관내피세포에서 항노화와 관련된 Sirt6 유전자의 발현을 촉진시키는 것을 확인함으로써, 4-헥실레조르시놀(4-hexylresorcinol)을 항노화 제제로 제공하는 것이다.The present invention provides an anti-aging composition, by confirming that 4-hexylresorcinol promotes the expression of the Sirt6 gene related to anti-aging in vascular endothelial cells, 4-hexylresorcinol (4) -hexylresorcinol) is provided as an anti-aging agent.
본 발명은 4-헥실레조르시놀(4-hexylresorcinol)을 유효성분으로 포함하는 항노화용 조성물을 제공한다.The present invention provides an anti-aging composition containing 4-hexylresorcinol as an active ingredient.
또한, 본 발명은 상기 항노화용 조성물을 유효성분으로 포함하는 노화 질환의 예방 또는 치료용 약학적 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing or treating aging diseases comprising the anti-aging composition as an active ingredient.
또한, 본 발명은 상기 항노화용 조성물을 유효성분으로 포함하는 노화 질환의 예방 또는 개선용 건강기능 식품 조성물을 제공한다.In addition, the present invention provides a health functional food composition for preventing or improving aging diseases containing the anti-aging composition as an active ingredient.
또한, 본 발명은 4-헥실레조르시놀(4-hexylresorcinol)을 유효성분으로 포함하는 Sirt6의 발현 촉진을 위한 시약 조성물을 제공한다.Additionally, the present invention provides a reagent composition for promoting the expression of Sirt6 containing 4-hexylresorcinol as an active ingredient.
본 발명에 따르면, 4-헥실레조르시놀(4-hexylresorcinol)이 혈관내피세포에서 항노화와 관련된 Sirt6 유전자의 발현을 촉진시키는 것을 확인함으로써, 4-헥실레조르시놀(4-hexylresorcinol)을 항노화 제제로 제공될 수 있다.According to the present invention, by confirming that 4-hexylresorcinol promotes the expression of the Sirt6 gene related to anti-aging in vascular endothelial cells, 4-hexylresorcinol is used as an anti-aging agent. It may be provided as a formulation.
도 1은 4-헥실레조르시놀(4-hexylresorcinol)이 Sirt6의 발현을 유도하여 항노화 기작을 나타내는 모식도이다.
도 2는 4-헥실레조르시놀(4-hexylresorcinol)이 Sirt6 유전자의 발현에 미치는 영향을 평가한 결과이다.
도 3은 4-헥실레조르시놀(4-hexylresorcinol)이 Sirtuin 6 단백질 발현에 미치는 영향을 평가한 결과이다.Figure 1 is a schematic diagram showing the anti-aging mechanism of 4-hexylresorcinol by inducing the expression of Sirt6.
Figure 2 shows the results of evaluating the effect of 4-hexylresorcinol on the expression of the Sirt6 gene.
Figure 3 shows the results of evaluating the effect of 4-hexylresorcinol on Sirtuin 6 protein expression.
본 명세서에서 사용되는 용어는 본 발명에서의 기능을 고려하면서 가능한 현재 널리 사용되는 일반적인 용어들을 선택하였으나, 이는 당 분야에 종사하는 기술자의 의도 또는 판례, 새로운 기술의 출현 등에 따라 달라질 수 있다. 또한, 특정한 경우는 출원인이 임의로 선정한 용어도 있으며, 이 경우 해당되는 발명의 설명 부분에서 상세히 그 의미를 기재할 것이다. 따라서 본 발명에서 사용되는 용어는 단순한 용어의 명칭이 아닌, 그 용어가 가지는 의미와 본 발명의 전반에 걸친 내용을 토대로 정의되어야 한다.The terms used in this specification are general terms that are currently widely used as much as possible while considering the function in the present invention, but this may vary depending on the intention or precedent of a person skilled in the art, the emergence of new technology, etc. In addition, in certain cases, there are terms arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the relevant invention. Therefore, the terms used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, rather than simply the name of the term.
다르게 정의되지 않는 한, 기술적이거나 과학적인 용어를 포함해서 여기서 사용되는 모든 용어들은 본 발명이 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 문맥상 가지는 의미와 일치하는 의미를 가지는 것으로 해석되어야 하며, 본 출원에서 명백하게 정의하지 않는 한, 이상적이거나 과도하게 형식적인 의미로 해석되지 않는다.Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the technical field to which the present invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and unless clearly defined in the present application, should not be interpreted in an ideal or excessively formal sense. No.
수치 범위는 상기 범위에 정의된 수치를 포함한다. 본 명세서에 걸쳐 주어진 모든 최대의 수치 제한은 낮은 수치 제한이 명확히 쓰여 있는 것처럼 모든 더 낮은 수치 제한을 포함한다. 본 명세서에 걸쳐 주어진 모든 최소의 수치 제한은 더 높은 수치 제한이 명확히 쓰여 있는 것처럼 모든 더 높은 수치 제한을 포함한다. 본 명세서에 걸쳐 주어진 모든 수치 제한은 더 좁은 수치 제한이 명확히 쓰여 있는 것처럼, 더 넓은 수치 범위 내의 더 좋은 모든 수치 범위를 포함할 것이다.The numerical range includes the values defined in the range above. Any maximum numerical limit given throughout this specification includes all lower numerical limits as if the lower numerical limit were explicitly written out. Every minimum numerical limit given throughout this specification includes every higher numerical limit as if such higher numerical limit were expressly written out. All numerical limits given throughout this specification will include all better numerical ranges within the broader numerical range, as if the narrower numerical limits were clearly written.
이하, 본 발명을 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명은 4-헥실레조르시놀(4-hexylresorcinol)을 유효성분으로 포함하는 항노화용 조성물을 제공한다.The present invention provides an anti-aging composition containing 4-hexylresorcinol as an active ingredient.
상기 4-헥실레조르시놀(4-hexylresorcinol)은 Sirt6의 발현을 촉진한다. 상기 Sirt6 유전자는 동물 모델에서 녹아웃시 조로증(Premature aging)의 증상이 나타나고, 과발현시 수명이 약 15-20% 증가하는 것으로 보고되었으며, 항노화와 수면연장에 관련이 있는 유전자로 보고되었다.The 4-hexylresorcinol promotes the expression of Sirt6. The Sirt6 gene has been reported to show symptoms of premature aging when knocked out in animal models, and to increase lifespan by about 15-20% when overexpressed, and has been reported to be a gene related to anti-aging and prolonged sleep.
또한, 본 발명은 상기 항노화용 조성물을 유효성분으로 포함하는 노화 질환의 예방 또는 치료용 약학적 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing or treating aging diseases comprising the anti-aging composition as an active ingredient.
상기 노화 질환은 시르투인 6(Sirtuin 6)의 결핍으로 발병되는 질환이고, 구체적으로 암, 지방간, 심근비대증, 조기 노화, 당뇨병, 알츠하이머병, 및 염증성 질환으로 이루어진 군에서 선택된 어느 하나일 수 있다.The aging disease is a disease caused by a deficiency of Sirtuin 6, and may specifically be any one selected from the group consisting of cancer, fatty liver, myocardial hypertrophy, premature aging, diabetes, Alzheimer's disease, and inflammatory disease. .
상기 약학적 조성물은 시르투인 6(Sirtuin 6)의 수준이 낮은 환자를 대상으로 하며, 시르투인 6(Sirtuin 6)의 수준이 낮은 환자로부터 시르투인 6(Sirtuin 6)의 결핍으로 발병되는 질환을 개선시키기 위해 투여될 수 있다.The pharmaceutical composition is intended for patients with low levels of Sirtuin 6, and is used to treat diseases caused by deficiency of Sirtuin 6 in patients with low levels of Sirtuin 6. It can be administered to improve a disease.
본 발명의 약학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다.The pharmaceutical composition of the present invention is prepared in unit dosage form or in multi-dose form by formulating it using a pharmaceutically acceptable carrier according to a method that can be easily performed by those skilled in the art. It can be manufactured by placing it in a container.
상기 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸 히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.The pharmaceutically acceptable carriers are those commonly used in preparation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, Includes, but is not limited to, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above ingredients, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc.
본 발명에 있어서, 상기 약학적 조성물에 포함되는 첨가제의 함량은 특별히 한정되는 것은 아니며 통상의 제제화에 사용되는 함량 범위 내에서 적절하게 조절될 수 있다.In the present invention, the content of additives included in the pharmaceutical composition is not particularly limited and can be appropriately adjusted within the content range used in conventional formulations.
상기 약학적 조성물은 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립, 정제, 크림, 젤, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 및 카타플라스마제로 이루어진 군으로부터 선택되는 하나 이상의 외용제 형태로 제형화될 수 있다.The pharmaceutical compositions include injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, tablets, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, and cataplasma It may be formulated in the form of one or more external preparations selected from the group consisting of:
본 발명의 약학적 조성물은 제형화를 위해 추가로 있는 약학적으로 허용가능한 담체 및 희석제를 포함할 수 있다. 상기 약학적으로 허용가능한 담체 및 희석제는 전분, 당, 및 만니톨과 같은 부형제, 칼슘 포스페이트 등과 같은 충전제 및 증량제, 카르복시메틸셀룰로오스, 히드록시프로필셀룰로오스 등과 같은 셀룰로오스 유도체, 젤라틴, 알긴산염, 및 폴리비닐 피롤리돈 등과 같은 결합제, 활석, 스테아린산 칼슘, 수소화 피마자유 및 폴리에틸렌 글리콜과 같은 윤활제, 포비돈, 크로스포비돈과 같은 붕해제, 폴리소르베이트, 세틸알코올, 및 글리세롤 등과 같은 계면활성제를 포함하나, 이에 한정되지 않는다. 상기 약학적으로 허용가능한 담체 및 희석제는 대상체에게 생물학적 및 생리학적으로 친화적인 것일 수 있다. 희석제의 예로는 염수, 수용성 완충액, 용매 및/또는 분산제(dispersion media)를 들 수 있으나, 이에 제한되는 것은 아니다.The pharmaceutical composition of the present invention may additionally contain pharmaceutically acceptable carriers and diluents for formulation. The pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar, and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose, hydroxypropylcellulose, gelatin, alginate, and polyvinyl chloride. Including, but not limited to, binders such as rolidone, lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone and crospovidone, and surfactants such as polysorbate, cetyl alcohol, and glycerol. No. The pharmaceutically acceptable carrier and diluent may be biologically and physiologically friendly to the subject. Examples of diluents include, but are not limited to, saline, aqueous buffers, solvents, and/or dispersion media.
본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구 투여(예를 들어, 정맥 내, 피하, 복강 내 또는 국소에 적용)할 수 있다. 경구 투여일 경우, 정제, 트로키제 (troches), 로젠지 (lozenge), 수용성 현탁액, 유성 현탁액, 조제 분말, 과립, 에멀젼, 하드 캡슐, 소프트 캡슐, 시럽 또는 엘릭시르제 등으로 제형화될 수 있다. 비경구 투여일 경우, 주사액, 좌제, 호흡기 흡입용 분말, 스프레이용 에어로졸제, 연고, 도포용 파우더, 오일, 크림 등으로 제형화 될 수 있다.The pharmaceutical composition of the present invention can be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally, or topically) depending on the desired method. For oral administration, it may be formulated as tablets, troches, lozenges, aqueous suspensions, oily suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups, or elixirs. In the case of parenteral administration, it can be formulated as an injection, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, cream, etc.
본 발명의 약학적 조성물의 투여량은 환자의 상태 및 체중, 연령, 성별, 건강상태, 식이 체질 특이성, 제제의 성질, 질병의 정도, 조성물의 투여시간, 투여방법, 투여기간 또는 간격, 배설율, 및 약물 형태에 따라 그 범위가 다양할 수 있으며, 이 분야 통상의 기술자에 의해 적절하게 선택될 수 있다. 예컨대, 약 0.1 내지 10,000 mg/kg의 범위일 수 있으나 이제 제한되지 않으며, 하루 일회 내지 수회에 나누어 투여될 수 있다.The dosage of the pharmaceutical composition of the present invention is determined by the patient's condition and weight, age, gender, health status, dietary constitution specificity, nature of the preparation, degree of disease, administration time of the composition, administration method, administration period or interval, and excretion rate. , and the range may vary depending on the drug form, and may be appropriately selected by a person skilled in the art. For example, it may range from about 0.1 to 10,000 mg/kg, but is not limited and may be administered once or in divided doses several times a day.
상기 약학적 조성물은 목적하는 방법에 따라 경구 투여되거나 비경구 투여(예를 들면, 정맥 내, 피하 내, 복강 내 또는 국소에 적용)될 수 있다. 본 발명의 약학적 조성물의 약학적 유효량, 유효 투여량은 약학적 조성물의 제제화 방법, 투여 방식, 투여 시간 및/또는 투여 경로 등에 의해 다양해질 수 있으며, 당해 기술 분야에서 통상의 지식을 가진 자는 목적하는 치료에 효과적인 투여량을 용이하게 결정하고 처방할 수 있다. 본 발명의 약학적 조성물의 투여는 하루에 1회 투여될 수 있고, 수회에 나누어 투여될 수도 있다.The pharmaceutical composition may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally, or topically) depending on the desired method. The pharmaceutically effective amount and effective dosage of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time, and/or administration route of the pharmaceutical composition, and those skilled in the art will understand the purpose. Effective dosages for treatment can be easily determined and prescribed. The pharmaceutical composition of the present invention may be administered once a day, or may be administered in several divided doses.
또한, 본 발명은 상기 항노화용 조성물을 유효성분으로 포함하는 노화 질환의 예방 또는 개선용 건강기능 식품 조성물을 제공한다.In addition, the present invention provides a health functional food composition for preventing or improving aging diseases comprising the anti-aging composition as an active ingredient.
본 발명은 통상적으로 이용되는 식품으로써 일반적으로 사용될 수 있다.The present invention can be generally used with commonly used foods.
본 발명의 식품 조성물은 건강기능식품으로서 사용될 수 있다. 상기 "건강기능 식품"이라 함은 건강기능 식품에 관한 법률에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.The food composition of the present invention can be used as a health functional food. The term “health functional food” refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body in accordance with the Health Functional Food Act, and “functional” refers to food that is related to the structure and function of the human body. It means ingestion for the purpose of controlling nutrients or obtaining useful health effects such as physiological effects.
본 발명의 식품 조성물은 통상의 식품 첨가물을 포함할 수 있으며, 상기 "식품 첨가물"로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전처에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The food composition of the present invention may contain common food additives, and its suitability as a “food additive” is determined in accordance with the general provisions and general test methods of the food additive code approved by the Ministry of Food and Drug Safety, unless otherwise specified. The decision is made based on the specifications and standards for the item.
상기 "식품 첨가물 공전"에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼륨, 니코틴산, 계피산 등의 화학적 합성물, 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물, L-글루타민산나트륨 제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류들을 들 수 있다.Items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as subchromic pigment, licorice extract, crystalline cellulose, high-liquid pigment, and guar gum; Examples include mixed preparations such as sodium L-glutamate preparations, noodle additive alkaline preparations, preservative preparations, and tar coloring preparations.
본 발명의 식품 조성물은 정제, 캡슐, 분말, 과립, 액상, 환 등의 형태로 제조 및 가공할 수 있다.The food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc.
예를 들어, 캡슐 형태의 건강기능 식품 중 경질캡슐제는 통상의 경질캡슐에 본 발명에 따른 조성물을 부형제 등의 첨가제와 혼합 및 충진하여 제조할 수 있으며, 연질캡슐제는 본 발명에 따른 조성물의 부형제 등의 첨가제와 혼합하고 젤라틴 등 캡슐기제에 충진하여 제조할 수 있다. 상기 연질캡슐제는 필요에 따라 글리세린 또는 소르비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다.For example, among health functional foods in the form of capsules, hard capsules can be manufactured by mixing and filling the composition according to the present invention with additives such as excipients in ordinary hard capsules, and soft capsules can be manufactured by mixing and filling the composition according to the present invention. It can be manufactured by mixing with additives such as excipients and filling it with a capsule base such as gelatin. The soft capsule may contain plasticizers such as glycerin or sorbitol, colorants, preservatives, etc., if necessary.
상기 부형제, 결합제, 붕해제, 활택제, 교미제, 착향제 등에 대한 용어 정의는 당업계에 공지된 문헌에 기재된 것으로 그 기능 등이 동일 내지 유사한 것들을 포함한다. 상기 식품의 종류에는 특별한 제한이 없으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다.Definitions of terms such as excipients, binders, disintegrants, lubricants, coagulants, flavoring agents, etc. are described in literature known in the art and include those with the same or similar functions. There is no particular limitation on the type of food, and it includes all health functional foods in the conventional sense.
본 발명에서 용어 “예방”이란 본 발명에 따른 조성물의 투여로 질환의 억제 또는 지연시키는 모든 행위를 말한다. 본 발명에서 용어 “치료”는 본 발명에 따른 조성물의 투여로 질환의 증세가 호전되거나 이롭게 변경하는 모든 행위를 말한다. 본 발명에서 "개선"이란 본 발명의 조성물을 개체에 투여하거나 섭취시켜 질환의 나쁜 상태를 좋게 하는 모든 행위를 의미한다.In the present invention, the term “prevention” refers to all actions that suppress or delay a disease by administering the composition according to the present invention. In the present invention, the term “treatment” refers to any action that improves or beneficially changes the symptoms of a disease by administering the composition according to the present invention. In the present invention, “improvement” means any action that improves the bad condition of a disease by administering or ingesting the composition of the present invention to an individual.
또한, 본 발명은 4-헥실레조르시놀(4-hexylresorcinol)을 유효성분으로 포함하는 Sirt6의 발현 촉진을 위한 시약 조성물을 제공한다.Additionally, the present invention provides a reagent composition for promoting the expression of Sirt6 containing 4-hexylresorcinol as an active ingredient.
이하, 본 발명의 이해를 돕기 위하여 실험예 및 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실험예 및 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실험예 및 실시예에 한정되는 것은 아니다. 본 발명의 실험예 및 실시예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.Hereinafter, to aid understanding of the present invention, it will be described in detail through experimental examples and examples. However, the following experimental examples and examples only illustrate the content of the present invention and the scope of the present invention is not limited to the following experimental examples and examples. Experimental examples and examples of the present invention are provided to more completely explain the present invention to those with average knowledge in the art.
<실험예> 실험 재료 및 방법<Experimental example> Experimental materials and methods
하기의 실험예들은 본 발명에 따른 각각의 실시예에 공통적으로 적용되는 실험예를 제공하기 위한 것이다.The following experimental examples are intended to provide experimental examples commonly applied to each embodiment according to the present invention.
1. 시약 및 기계1. Reagents and instruments
Fetal bovine serum (FBS)는 GIBCO (Grand Island, NY, U.S.A), oligo(dT)는 Promega (Madison, WI, USA)에서 구입하고, penicillin/streptomycin는 Lonza (Rockland, ME, USA)에서 구입하고, SuperScript Ⅱ Reverse Transcriptase는 Intron (iNtRON Biotechnology, Gyeinggi-do, Korea)에서 구입하고, SYBR green master mixture는 ABI (Carlsbad, CA, U.S.A)에서 구입하였다.Fetal bovine serum (FBS) was purchased from GIBCO (Grand Island, NY, U.S.A.), oligo(dT) was purchased from Promega (Madison, WI, USA), and penicillin/streptomycin was purchased from Lonza (Rockland, ME, USA). SuperScript Ⅱ Reverse Transcriptase was purchased from Intron (iNtRON Biotechnology, Gyeinggi-do, Korea), and SYBR green master mixture was purchased from ABI (Carlsbad, CA, U.S.A.).
Real-time PCR은 ViiA7 (Applied Biosystems, CA, U.S.A)을 사용하였고, DNA 및 RNA 정량은 Nano drop 2000, Incubator는 Thermo scientific(Rockford, IL, U.S.A)을 사용하였다. Protein 정량는 ELISA reader (TECAN, Mannedorf, Switzerland)를 사용하였다.Real-time PCR used ViiA7 (Applied Biosystems, CA, U.S.A.), Nano drop 2000 was used for DNA and RNA quantification, and Thermo scientific (Rockford, IL, U.S.A.) was used as an incubator. Protein quantification was performed using an ELISA reader (TECAN, Mannedorf, Switzerland).
2. HUVEC 배양2. HUVEC culture
HUVECs (Lonza, Walkersville, MD, USA)를 구입하여 1 μg/mL 하이드로 코르티손(hydrocortison), 12 μg/mL 소 뇌 추출물 (bovine brain extract), 50 μg/mL 젠타마이신 (gentamicin), 50 ng/mL 암포테리신-B (amphotericin-B), 10 ng/mL 표피성장인자(EGF), VEGF, FGF-2, 헤파린, 아스코르브산 및 10% 우태아혈청 (EGMTM-2, Clonetics®, Lonza, Walkersville, MD, USA)을 포함하는 내피 세포 성장 배지 -2 (Clonetics®, Lonza)에서 배양하였다. 세포는 37.5℃, 5% CO2 인큐베이터에서 배양되었으며, 마이코플라스마가 없는 세포 만 분석되었는지 확인하기 위해 정기적으로 마이코플라스마 assay를 이용하여 검증하였다.HUVECs (Lonza, Walkersville, MD, USA) were purchased and incubated with 1 μg/mL hydrocortisone, 12 μg/mL bovine brain extract, 50 μg/mL gentamicin, and 50 ng/mL. Amphotericin-B, 10 ng/mL epidermal growth factor (EGF), VEGF, FGF-2, heparin, ascorbic acid, and 10% fetal bovine serum (EGMTM-2, Clonetics®, Lonza, Walkersville, MD, USA) were cultured in endothelial cell growth medium-2 (Clonetics®, Lonza) containing Cells were cultured in an incubator at 37.5°C and 5% CO2 , and were regularly verified using a mycoplasma assay to ensure that only cells without mycoplasma were analyzed.
3. Real-time PCR3. Real-time PCR
Sirt6의 유전자 발현변화를 확인하기 위해 Real-time PCR을 수행하였다. 4-헥실레조르시놀(4-hexylresorcinol, 4-HR)을 농도별로 0, 1, 10, 100 μM씩 처리하고 24 시간 뒤 Total RNA를 추출하여, 1.5 ㎍의 Total RNA을 역전사효소를 이용하여 cDNA로 합성하였다. 합성된 cDNA 4 ㎕ (1/40 dilution)에 2× SYBR green PCR master mixture (5 ㎕)와 specific primer (0.5 ㎕)를 섞어서 real-time PCR을 수행하였다. Real-time PCR에 사용한 primer 세트(F: TGTGCCAAGTGTAAGACGCA, 서열번호 1; 및 R: CTTGGCCTTCACCCTTTTG, 서열번호 2)를 사용하였다.Real-time PCR was performed to confirm changes in gene expression of Sirt6. 4-hexylresorcinol (4-HR) was treated at each concentration of 0, 1, 10, and 100 μM, and total RNA was extracted 24 hours later, and 1.5 μg of total RNA was converted to cDNA using reverse transcriptase. It was synthesized. Real-time PCR was performed by mixing 4 ㎕ (1/40 dilution) of the synthesized cDNA with 2× SYBR green PCR master mixture (5 ㎕) and specific primer (0.5 ㎕). The primer set used for real-time PCR (F: TGTGCCAAGTGTAAGACGCA, SEQ ID NO: 1; and R: CTTGGCCTTCACCCTTTTG, SEQ ID NO: 2) was used.
4. Western blot4. Western blot
Sirt 6의 단백질 발현변화를 확인하기 위해 HUVEC 세포수가 70%정도 일 때 4-HR을 0, 1, 10, 100 μM로 처리하여 2, 8, 24 시간 뒤에 단백질을 추출하였다. 단백질 정량을 하고 sirt 6 항체를 (Abcam, Cambridge, UK)로 western blot을 시행하여 sirt 6의 단백질 발현을 확인하였다.To confirm changes in protein expression of Sirt 6, when the number of HUVEC cells was about 70%, 4-HR was treated with 0, 1, 10, and 100 μM, and protein was extracted 2, 8, and 24 hours later. Protein quantification was performed and western blot was performed using sirt 6 antibody (Abcam, Cambridge, UK) to confirm protein expression of sirt 6.
실시예: 4-HR이 Sirt6의 발현에 미치는 영향 평가Example: Evaluation of the effect of 4-HR on the expression of Sirt6
4-헥실레조르시놀(4-hexylresorcinol)이 항노화 및 수면연장 인자인 Sirt6 유전자의 발현에 미치는 영향을 평가하였다. 도 2 및 3에 나타난 바와 같이, 4-헥실레조르시놀(4-hexylresorcinol)이 인간 혈관내피세포(HUVECs)에서 Sirt6 유전자의 발현을 촉진할 뿐만 아니라, Sirtuin 6 단백질 발현을 촉진하는 것으로 확인되었다. 상기 결과는 항노화 및 건강한 수면연장과 관련된 Sirt6 유전자의 발현을 4-헥실레조르시놀(4-hexylresorcinol)이 촉진하는 것을 입증하며, 4-헥실레조르시놀(4-hexylresorcinol)이 항노화제로 사용될 수 있음을 입증한다.The effect of 4-hexylresorcinol on the expression of the Sirt6 gene, an anti-aging and sleep prolonging factor, was evaluated. As shown in Figures 2 and 3, it was confirmed that 4-hexylresorcinol not only promotes the expression of the Sirt6 gene in human vascular endothelial cells (HUVECs), but also promotes the expression of Sirtuin 6 protein. The above results demonstrate that 4-hexylresorcinol promotes the expression of the Sirt6 gene related to anti-aging and healthy sleep extension, and that 4-hexylresorcinol can be used as an anti-aging agent. Prove that you can.
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 즉, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다.As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. do. That is, the practical scope of the present invention is defined by the appended claims and their equivalents.
Claims (8)
A reagent composition for promoting the expression of Sirt6 containing 4-hexylresorcinol as an active ingredient.
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