KR101996383B1 - An anticancer composition comprising purtiton - Google Patents
An anticancer composition comprising purtiton Download PDFInfo
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- KR101996383B1 KR101996383B1 KR1020180086173A KR20180086173A KR101996383B1 KR 101996383 B1 KR101996383 B1 KR 101996383B1 KR 1020180086173 A KR1020180086173 A KR 1020180086173A KR 20180086173 A KR20180086173 A KR 20180086173A KR 101996383 B1 KR101996383 B1 KR 101996383B1
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- puritone
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Abstract
본 발명은 퓨리톤(puriton)을 포함하는 항암용 조성물에 관한 것으로, 본 발명의 퓨리톤은 인간 유방암 세포주 MDA-MB-231, 인간 간암 세포주 HepG2, 인간 폐암 세포주 A549, 인간 난소암 세포주 OVCA 429, 인간 혈액암 세포주 HL60, 인간 피부암 세포주 A375, 인간 신장암 세포주 RCC4(-) 및 인간 대장암 세포주 HCT116의 암세포 증식을 억제할 수 있으므로 항암용 조성물로서 유용하게 사용할 수 있다. 또한 본 발명의 퓨리톤은 세포 독성이 없어 약학적 및 식품 조성물에 안전하게 사용할 수 있다. The present invention relates to an anticancer composition comprising puritone, wherein the puritone of the present invention is a human breast cancer cell line MDA-MB-231, a human liver cancer cell line HepG2, a human lung cancer cell line A549, a human ovarian cancer cell line OVCA 429, Human cancer cell line HL60, human skin cancer cell line A375, human renal cancer cell line RCC4 (-), and human colon cancer cell line HCT116, and thus can be usefully used as an anticancer composition. In addition, the puritone of the present invention is not cytotoxic and can be safely used in pharmaceutical and food compositions.
Description
본 발명은 퓨리톤(puriton)을 포함하는 항암용 조성물에 관한 것으로, 보다 구체적으로는 퓨리톤을 유효성분으로 함유하는 항암용 약학적 조성물 및 식품 조성물에 관한 것이다. The present invention relates to an anticancer composition containing puritone, and more particularly to a pharmaceutical composition and a food composition for anticancer which contain puritone as an active ingredient.
항암제는 악성종양을 치료하기 위해 사용하는 방법들 중 수술이나 방사선 요법을 제외한 화학요법제를 총칭하는 것으로, 주로 핵산의 합성을 저해하여 항암활성을 나타내는 물질이 대부분이다. 그 예로, 파클리탁셀(paclitaxel), 도세탁셀(docetaxel) 등의 화합물은 미국 식품의약국의 승인을 받아 실제 임상에서 사용되고 있으며, 세포의 미세소관을 과도하게 안정화하여 정상적인 세포 분열을 억제함으로써 그 효과를 나타낸다. 하지만, 이들 항암제는 암세포 외에도 정상세포, 특히 세포분열이 활발한 정상 조직에도 손상을 입혀 위장관 점막 손상, 탈모, 복통 등의 부작용을 나타내며, 약물 자체의 심각한 독성뿐만 아니라 수용액 상에서의 낮은 용해도로 인하여 그 치료효능이 반감되는 결과를 가져온다. 따라서, 이러한 약물의 부작용을 최소화하고 암세포에서 효과를 극대화할 수 있는 새로운 암 치료제의 개발이 요구되고 있다.Among chemotherapeutic agents used for treating malignant tumors, chemotherapeutic agents other than surgery and radiation therapy are mainly used, and most of them exhibit anticancer activity by inhibiting the synthesis of nucleic acids. For example, paclitaxel, docetaxel, and other compounds have been approved by the US Food and Drug Administration (EPA) for use in clinical practice and have shown effects by over-stabilizing the microtubules of cells and inhibiting normal cell division. However, these anticancer drugs are not only cancer cells but also normal cells, especially cell division-active normal tissues, resulting in adverse effects such as gastrointestinal mucosal damage, hair loss, abdominal pain and the like. In addition to the serious toxicity of the drug itself and low solubility in the aqueous solution, Resulting in half the efficacy. Therefore, there is a need to develop a novel cancer treatment agent that can minimize the side effects of such drugs and maximize the effects of the cancer cells.
유방암은 미국을 비롯한 서구 국가에서 발병률이 가장 높은 암으로 한국을 포함한 동양인에서는 발생이 다소 낮은 편이었지만, 식생활의 변화와 환경의 변화로 인해 한국인 여성에서의 발생률은 매년 증가하고 있는 추세이다.Breast cancer has the highest incidence rate in Western countries including the United States, and it has been reported to be slightly lower in Asians, including Koreans. However, the incidence in Korean women is increasing every year due to changes in diet and environment.
유방암을 일으키는 여러 가지 요인으로는 연령, 이른 초경, 늦은 폐경, 신장, 폐경 후 비만, 유방암 가족력, 방사선 노출, 경구피임약, 호르몬 대체 치료법, 유방 조직의 밀도, 유전자 변이 및 유방의 양성종양과 같은 요인들이 유방암의 발생 위험을 증가시키며 그 중에서도 식생활에 의한 유방암 발생이 전체 유방암 발생률 중 약 35%를 차지한다고 알려져 있다.Factors such as age, early menarche, late menopause, kidney, postmenopausal obesity, family history of breast cancer, radiation exposure, oral contraceptive, hormone replacement therapy, density of breast tissue, genetic variation and benign tumors of breast Are associated with increased risk of breast cancer, among which dietary breast cancer is known to account for about 35% of all breast cancer incidences.
한편, 퓨리톤은 20여년전부터 광물의학을 토대로 미국과 한국에서 연구개발된 100% 광물성 천연물질로서 질병치료와 피부건강 및 건강의 밸런스를 유지시키는 제품이다. US FDA로부터 성분분석, 독성분석, 영양분석을 통과해서 인체내(눈, 간, 피부) 모든 부분에 안전성을 인정받았으며, 일반의약품(NDC)으로 승인을 받았다.On the other hand, Puritone is a 100% mineral natural substance that has been research and developed in the US and Korea based on mineral medicine since 20 years ago, and it is a product that maintains balance of disease treatment and skin health and health. It has been approved by the US Food and Drug Administration (FDA) for its safety in all parts of the body (eye, liver, skin) and for its approval as a generic medicinal product (NDC) through its component analysis, toxicity analysis and nutritional analysis.
현재 출시한 제품은 5가지의 일반 의약품이 있다. 퓨리톤 안약(Puriton Eye Relief Drop), 얼굴치료, 피부치료, 화상 절상 등 응급처치, 여성 청결치료제 등이다. Currently, there are 5 generic drugs. Puriton Eye Relief Drop, facial treatment, skin treatment, first-aid treatment such as burn-up, and women's clean treatment.
퓨리톤 안약(Puriton Eye Drop)은 눈건조증에 대해 높은 치료효능을 보일 뿐만 아니라 불치에 가까운 눈질환인 백내장, 녹내장, 비문증 등의 질환치료에 이르기까지 효능을 나타낸다고 알려졌다. 또한, 퓨리톤 여성 청결치료제(puriton Intimate Disinfection Spray)는 대부분 여성의 깊은 고민으로 자리하고 있는 질염 및 방광염과 일반적 치질에 이르기까지 그 질환의 균을 완벽하게 잡아줌으로써 극대의 치료효능을 가진 신개념 의약품이라 할 수 있다.Puriton Eye Drop is known to have high therapeutic efficacy against dry eye syndrome, and is effective in treating diseases like cataracts, glaucoma, and insomnia, which are close to incurable diseases. In addition, Puriton Intimate Disinfection Spray is a new concept medicine with maximal therapeutic efficacy by completely capturing the bacteria of the disease from vaginitis and cystitis which are mostly women's deep troubles and general hemorrhoids. can do.
이에 본 발명자들은 퓨리톤의 알려진 효능 이외에 새로운 효능을 연구하던 중 퓨리톤이 암세포 증식을 억제시킬 수 있음을 발견하고 본 발명을 완성하였다.Accordingly, the inventors of the present invention discovered that puritone can inhibit cancer cell proliferation while studying new efficacy in addition to the known efficacy of puritone, and completed the present invention.
따라서, 본 발명에서 해결하고자 하는 기술적 과제는 항암효과를 가지는 신규 물질을 유효성분으로 포함하는 항암용 조성물을 제공하기 위한 것이다.Accordingly, an object of the present invention is to provide an anticancer composition comprising a novel substance having an anticancer effect as an active ingredient.
또한, 본 발명에서 해결하고자 하는 다른 기술적 과제는 상기 항암용 조성물을 개체에 투여하여 암을 예방, 개선 또는 치료하는 방법을 제공하기 위한 것이다.Another object of the present invention is to provide a method for preventing, ameliorating or treating cancer by administering the composition for cancer therapy to a subject.
상기한 기술적 과제를 해결하기 위하여, 본 발명에서는 퓨리톤을 유효성분으로 포함하는 항암용 조성물을 제공한다.In order to solve the above-mentioned technical problems, the present invention provides an anticancer composition comprising puritone as an active ingredient.
바람직하게, 본 발명에서 퓨리톤을 포함하는 조성물은 약학적 조성물 또는 식품 조성물일 수 있다.Preferably, the composition comprising the puritone in the present invention may be a pharmaceutical composition or a food composition.
또한, 본 발명에서는 상기한 다른 기술적 과제를 해결하기 위하여, 퓨리톤을 유효성분으로 포함하는 항암용 조성물을 개체에 투여하여 암을 예방, 개선 또는 치료하는 방법을 제공한다.The present invention also provides a method of preventing, ameliorating or treating cancer by administering to a subject an anti-cancer composition comprising puritone as an active ingredient.
본 발명에서 암은 갑상선암, 위암, 대장암, 폐암, 난소암, 유방암, 직장암, 간암, 전립선암, 췌장암, 담낭암, 혈액암, 피부암, 신장암 또는 담도암을 예로 들 수 있으나, 반드시 이로 한정되는 것은 아니다.In the present invention, the cancer is exemplified by thyroid cancer, stomach cancer, colon cancer, lung cancer, ovarian cancer, breast cancer, rectal cancer, liver cancer, prostate cancer, pancreatic cancer, gallbladder cancer, blood cancer, skin cancer, kidney cancer or bile duct cancer. It is not.
본 발명에서 항암 조성물의 유효성분인 퓨리톤은 미네랄 이온을 함유하는 액상형태로서 시판되는 것을 구입하여 사용하거나 직접 제조하여 사용할 수 있다. 예를 들어, 퓨리톤은 천연광물로 점토계 원료인 고령토, 벤토나이트, 제올라이트, 탄산칼슘, 운모류 등의 광물을 이용하여 제조될 수 있는데,이들 광물은 분쇄전 900℃ 이상의 고온소성을 거쳐 원료내에 함유된 유해 불순물 및 중금속 등을 제거하는 공정을 거친다. 소성공정을 마친 원료를 분쇄공정을 통해 원하는 미세한 나노입자의 광물로 가공처리를 하게 된다. 상기 분쇄는 건식 또는 습식 또는 이들을 순차적으로 혼합한 방식으로 이루어질 수 있다. 건식공정에서는 정밀한 미세입자의 생산성은 낮으며 단계별로 미세한 입자를 얻을 수 있으나 대량적인 생산에 한계가 있고 정밀성이 떨어진다. 반면 습식공정은 준비된 광물을 볼밀에 준비된 광물을 투입하여 적당량의 물을 투입하여 고속의 회전을 가하여 미분말을 얻을 수 있는 아주 미세하고 정밀한 입자를 대량적으로 생산할 수 있으나 제조비용이 건식에 비해 높은 단점이 있다. 일반적으로 퓨리톤 액상 제조에 투입되는 광물의 입자크기는 나노입자에 해당되는 5000 mesh(2㎛ 이하)이상의 아주 미세한 분말을 사용한다.In the present invention, the puritone, which is an effective component of the anticancer composition, can be used by purchasing a commercially available liquid form containing mineral ions or by directly preparing it. For example, purite is a natural mineral, which can be produced by using minerals such as kaolinite, bentonite, zeolite, calcium carbonate, and mica, which are clay materials. These minerals are calcined at a high temperature The harmful impurities and heavy metals are removed. After the calcination process, the raw material is subjected to a grinding process to process the desired fine nano-particle minerals. The pulverization may be carried out in a dry or wet process or a sequential mixing process. In the dry process, the productivity of fine fine particles is low, and fine particles can be obtained in stages, but there is a limit in mass production and precision is poor. On the other hand, the wet process can produce very fine and precise particles that can obtain fine powder by applying a suitable amount of water to the ball mill, . In general, the particle size of the minerals used for the preparation of the purite liquid phase is very fine powder of 5000 mesh (2 μm or less) corresponding to the nanoparticles.
습식분쇄된 나노입자의 미분말 광물질을 고온 고압 공정장치에 적당량의 광물분말과 증류수를 투입하여 밀폐된 개방형 혼합기에 투입하여 수행한다. 100-300℃ 이상의 열을 가하여 투입된 광물과 물이 고온에서 반응하여 인체에 필수적인 이온화된 미네랄을 용출시키는 공정이며 적정 가열시간을 부여하여 인체에 필요한 다량의 광물성 미네랄을 얻게 된다. 미네랄 용출공정을 거친 후 숙성공정인 광물의 침전과정을 거친다. 그 과정의 부여시간은 액상내에 이온화된 광물의 입자크기에 영향을 미친다. 대략적으로 숙성공정은 원하는 제품에 따라 24시간에서 일주일 이상의 숙성시간이 필요하다.The pulverized minerals of the wet-pulverized nanoparticles are put into a closed open type mixer by putting an appropriate amount of mineral powder and distilled water into a high-temperature and high-pressure processing apparatus. It is a process of dissolving ionized minerals that are essential to the human body by reacting at a temperature of 100-300 ° C or higher with a mineral introduced at a high temperature, and giving a proper amount of heating time to obtain a large amount of mineral minerals required for the human body. Minerals are extracted and then subjected to a mineral precipitation process. The duration of the process affects the particle size of the ionized minerals in the liquid phase. Approximately, the aging process requires aging time of 24 hours to a week or more depending on the desired product.
숙성과정이 마무리 되면 콜로이드 액상과 침전된 광물의 미분말을 분리하는 필터링 공정을 거친다. 정밀한 필터링 공정을 통해 분리된 퓨리톤 미네랄 수용액을 제조할 수 있다.When the aging process is completed, a filtering process is performed to separate the colloidal liquid phase and the fine powder of the precipitated minerals. A precise filtering process can be used to produce an aqueous solution of the separated purite minerals.
퓨리톤의 원료는 식품 및 의약품의 용도로서 수많은 종류의 콜로이드성 미네랄이 함유된 몬모릴로나이트(Montmorillonites) 또는 벤토나이트(Bentonite) 등을 주요 구성물로 하는 안정된 점토광물을 주요성분 원료로 사용한다. 점토광물의 주원소는 Si, Al, Na, K, Ti이며 미량원소는 많은 원소 가운데 Fe, Ca, S, Mg, Ce, La, Zr, P 등이며 바이오세라믹 분말은 Fe2O3, TiO2, Na2O, SiO2, P2O5, CeO2, K2O, MgO 등이다. 제품의 특성은 몬모릴로나이트(또는 벤토나이트)인 점토광물을 미세한 입자크기로 분쇄한 후 콜로이드 상태로 정제처리 한 수많은 Trace minerals(미량원소)이 함유된 복합물질이다. 그리고 혈액세포보다 입자크기가 훨씬 작은 천연상태(Colloidal Trace Minerals)의 미네랄이다. 종합하면 점토광물로서 의학적 광물학(Medical Mineralogy)에서 인정된 점토광물이 퓨리톤의 주원료이다. The raw materials of puritone are mainly used as raw materials for food and pharmaceuticals, and stabilized clay minerals containing montmorillonites or bentonite, which contain many kinds of colloidal minerals. The major elements of clay minerals are Si, Al, Na, K, and Ti, and trace elements are Fe, Ca, S, Mg, Ce, La, Zr, and P. Bioceramic powders are Fe 2 O 3 , TiO 2 a, Na 2 O, SiO 2, P 2 O5,
상기 퓨리톤은 카올리나이트(Kaolinite), 몬트모릴로나이트(Montmorillonite), 바이오타이트(Biotite), 서펜틴(Serpentine) 및 마이카(Mica)를 필수적으로 포함하며, 바람직하게는 추가로 버미쿨라이트(Vermiculite), 무스코바이트(Muscovite), 클리노클로레(Clinochlore), 브루사이트(Brucite), 일라이트(Illite), 리메스톤(Limestone), 제올라이트(Zeolite) 및 올소클레이스(Orthoclase) 또는 올소클레이스 펠드스파(Orthoclase feldspar)를 포함할 수 있다.The puritone essentially comprises Kaolinite, Montmorillonite, Biotite, Serpentine and Mica, preferably further comprising Vermiculite, Clostridium, Muscovite, Clinochlore, Brucite, Illite, Limestone, Zeolite and Orthoclase or Oloclaysfeld (Orthoclase feldspar).
더욱 바람직하게, 퓨리톤은 미량의 추가의 성분을 포함할 수 있는데, 구체적으로 베마이트(boehmite), 네펠린(Nepheline), 팔리코르스카이트(Palygorskite) 또는 애터펄자이트(Attapulgite), 마그네사이트(Magnesite), 스페릴라이트(Sperrylite), 크렌네라이트(Krennerite), 페트자이트(Petzite), 기브사이트(Gibbsite), 앨러페인(Allophane), 플로고파이트(Phlogopite), 레피도라이트(Lepidolite), 실바나이트(Sylvanite), 글로코나이트(Glauconite), 돌로마이트(Dolomite), 토르말린(Tourmaline), 카날라이트(Carnallite), 안드라다이트(Andradite), 휴마이트(Humite)를 포함할 수 있다.More preferably, the puritone may comprise minor amounts of additional components, such as boehmite, Nepheline, Palygorskite or Attapulgite, magnesite Magnesite, Sperrylite, Krennerite, Petzite, Gibbsite, Allophane, Phlogopite, Lepidolite, And may include Sylvanite, Glauconite, Dolomite, Tourmaline, Carnallite, Andradite, Humite, and the like.
퓨리톤을 구성하는 구체적인 성분의 함량은 하기 표 1(단위 중량%)에 나타낸 바와 같다. The content of specific components constituting the purite is as shown in Table 1 (unit weight%).
본 발명의 조성물은 조성물의 총 중량을 기준으로 하여 퓨리톤이 0.005 내지 50 중량%, 보다 바람직하게는 0.01 내지 30 중량%, 가장 바람직하게는 0.1 내지 10 중량%로 포함할 수 있다. 이 때 퓨리톤의 함량이 0.005 중량% 미만일 경우 본 발명의 목적효과인 암세포 증식 억제효과를 수득할 수 없으며, 50 중량%를 초과할 경우 함량의 증가에 따라 효과가 비례적이지 않아 비효율적일 수 있으며 제형상의 안정성이 확보되지 않는 문제점이 있다.The composition of the present invention may contain 0.005 to 50 wt%, more preferably 0.01 to 30 wt%, and most preferably 0.1 to 10 wt% of furitone based on the total weight of the composition. If the content of puritone is less than 0.005% by weight, the cancer cell proliferation inhibiting effect of the present invention can not be obtained. If the content of puritone is more than 50% by weight, the effect may not be proportional to the content, There is a problem that the stability of the shape is not ensured.
본 발명의 퓨리톤을 함유하는 조성물은 암세포주의 증식 억제능을 가지며, 천연 물질로서 인체에 안전하다. The composition containing the puritone of the present invention has the ability to inhibit cancer cell proliferation and is safe for human body as a natural substance.
상기 약학적 조성물은 정제, 환제, 산제, 과립제, 캡슐제, 현탁제, 내용액제, 유제, 시럽제, 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결 건조제 및 좌제로 이루어진 군으로부터 선택되는 어느 하나의 제형을 가질 수 있으며, 경구 또는 비경구의 여러 가지 제형일 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용된다. 경구투여를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.The pharmaceutical composition may be any one selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, nonaqueous solvents, suspensions, emulsions, And may be oral or parenteral formulations of various forms. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.
본 발명의 조성물은 약학적으로 유효한 양으로 투여할 수 있다. 본 발명에서 용어 "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 개체 종류 및 중증도, 연령, 성별, 질병의 종류, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와 순차적 또는 동시에 투여될 수 있다. 그리고 단일 또는 다중 투여될 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 당업자에 의해 용이하게 결정될 수 있다. 본 발명의 조성물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물 형태, 투여경로 및 기간에 따라 다르며, 적합한 총 1일 사용량은 올바른 의학적 판단범위 내에서 처치의에 의해 결정될 수 있으나, 일반적으로 0.001 내지 1000 mg/kg의 양, 바람직하게는 0.05 내지 200 mg/kg, 보다 바람직하게는 0.1 내지 100 mg/kg의 양을 일일 1회 내지 수회로 나누어 투여할 수 있다. 상기 조성물은 항암 목적으로 하는 개체이면 특별히 한정되지 않고, 어떠한 개체이든 적용가능하다. 예를 들면, 원숭이, 개, 고양이, 토끼, 모르모트, 랫트, 마우스, 소, 양, 돼지, 염소 등과 같은 비인간동물, 인간, 조류 및 어류 등 어느 개체에나 적용할 수 있으며, 투여의 방식은 당업계의 통상적인 방법이라면 제한없이 포함한다. 예를 들어, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관 내 주사에 의해 투여될 수 있다.The composition of the present invention may be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will vary depending on the species and severity, age, sex, , The activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention 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 adverse effect, and can be easily determined by those skilled in the art. The preferred dosage of the composition of the present invention will depend on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, and the appropriate total daily dose may be determined by treatment, Generally, an amount of 0.001 to 1000 mg / kg, preferably 0.05 to 200 mg / kg, more preferably 0.1 to 100 mg / kg, can be administered in a single dose, divided into several times a day. The composition is not particularly limited as long as it is an object for anticancer purposes, and any object can be applied. For example, it can be applied to any non-human animal such as a monkey, a dog, a cat, a rabbit, a guinea pig, a rat, a mouse, a cattle, a sheep, a pig, Including but not limited to, For example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.
본 발명의 구체적인 실시양태에 따르면, 퓨리톤은 인간 유방암 세포주 MDA-MB-231, 인간 간암 세포주 HepG2, 인간 폐암 세포주 A549, 인간 난소암 세포주 OVCA 429, 인간 혈액암 세포주 HL60, 인간 피부암 세포주 A375, 인간 신장암 세포주 RCC4(-) 및 인간 대장암 세포주 HCT116에 대한 증식 억제효과를 나타내는 것을 확인할 수 있다. According to a specific embodiment of the present invention, the puritone is selected from human breast cancer cell line MDA-MB-231, human liver cancer cell line HepG2, human lung cancer cell line A549, human ovarian cancer
본 발명의 하나의 구체적인 구현예로서, 본 발명은 퓨리톤을 유효성분으로 함유하는, 항암용 식품 조성물을 제공한다.As one specific embodiment of the present invention, the present invention provides a food composition for anticancer, which contains puritone as an active ingredient.
구체적으로, 본 발명의 퓨리톤을 항암을 목적으로 식품 조성물에 포함시킬 수 있다. Specifically, the puritone of the present invention can be incorporated into a food composition for the purpose of anti-cancer.
본 발명에서 용어, "개선"은 상기 조성물을 이용하여 암의 의심 및 발병 개체의 증상이 호전되거나 이롭게 되는 모든 행위를 말한다.In the present invention, the term "improvement" refers to any action that alleviates or alleviates the suspicion of a cancer and symptoms of an onset entity using the composition.
본 발명의 조성물을 식품에 포함하여 사용할 경우, 상기 조성물을 그대로 첨가하거나 다른 건강식품 및 건강기능식품 또는 건강기능식품 성분과 함께 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용할 수 있다. 유효성분의 혼합량은 사용 목적에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조 시에 본 발명의 조성물은 원료에 대하여 바람직하게는 15 중량부 이하, 보다 바람직하게는 10 중량부 이하의 양으로 첨가할 수 있다. 그러나, 건강 조절 및 위생을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안정성 면에서 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용할 수 있다.When the composition of the present invention is used as a food, the composition may be added as it is or may be used in combination with other health food, health functional food or health functional food component, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use. Generally, the composition of the present invention may be added in an amount of preferably not more than 15 parts by weight, more preferably not more than 10 parts by weight, based on the raw material, in the production of food or beverage. However, in the case of long-term intake intended for health control and hygiene, the amount may be less than the above range, and since there is no problem in terms of stability, the active ingredient may be used in an amount in the above range.
본 발명의 조성물을 포함할 수 있는 식품의 종류에는 특별한 제한은 없으며, 구체적인 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있고, 통상적인 의미에서의 식품을 모두 포함할 수 있으며, 동물을 위한 사료로 이용되는 식품을 포함할 수 있다. 또한, 본 발명의 식품 조성물이 음료의 형태로 사용될 경우에는 통상의 음료와 같이 여러 가지 감미제, 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상기 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 수크로스와 같은 디사카라이드, 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 및 자일리톨, 소르비톨, 에리트리톨과 같은 당알콜일 수 있다. 상기 천연 탄수화물의 비율은 이에 제한되지는 않으나, 본 발명의 조성물 100 ㎖ 당 바람직하게는 약 0.01 내지 0.04 g, 보다 바람직하게는 0.02 내지 0.03 g일 수 있다. 상기 감미제는 타우마틴, 스테비아 추출물과 같은 천연 감미제 및 사카린, 아스파르탐과 같은 합성 감미제일 수 있다. 상기 외에 본 발명의 식품 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다.The composition of the present invention is not particularly limited, and examples thereof include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramie noodle, other noodles, gum, ice cream , Various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and may include all foods in the conventional sense, and foods used as feed for animals. In addition, when the food composition of the present invention is used in the form of a drink, it may contain various sweetening agents, flavoring agents, or natural carbohydrates as additional components such as ordinary beverages. The natural carbohydrates may be polysaccharides such as disaccharides such as monosaccharides such as glucose and fructose, maltose, sucrose, dextrin, cyclodextrins, and sugar alcohols such as xylitol, sorbitol and erythritol. The ratio of the natural carbohydrate is not limited thereto, but may be about 0.01 to 0.04 g, and more preferably 0.02 to 0.03 g per 100 ml of the composition of the present invention. The sweeteners may be natural sweeteners such as tau martin and stevia extract, and synthetic sweeteners such as saccharin and aspartame. In addition to the above, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , A carbonating agent used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks.
또 하나의 양태로서, 본 발명은 상기 약학적 조성물을 암의 의심개체에 투여하는 단계를 포함하는, 항암 방법을 제공한다. 본 발명에서 상기 암의 의심 개체는 암이 발병하였거나 발병할 수 있는 인간을 포함한 모든 동물을 의미하며, 본 발명의 화합물 또는 이의 약학적으로 허용가능한 염을 포함하는 약학적 조성물을 암의 의심 개체에 투여함으로써, 개체를 효율적으로 치료할 수 있다. In another aspect, the present invention provides an anticancer method comprising the step of administering the pharmaceutical composition to a suspected individual of a cancer. In the present invention, the suspected individual of the cancer refers to all animals including humans that can develop or develop cancer, and a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof may be administered to a suspect subject By administering, the individual can be treated efficiently.
본 발명에서 용어 "투여"는 어떠한 적절한 방법으로 암 의심 개체에게 본 발명의 약학적 조성물을 도입하는 것을 의미하며, 투여 경로는 목적 조직에 도달할 수 있는 한 경구 또는 비경구의 다양한 경로를 통하여 투여될 수 있다. 본 발명의 치료 방법은 상기 퓨리톤을 포함하는 약학적 조성물을 약학적 유효량으로 투여하는 것을 포함할 수 있다. 적합한 총 1일 사용량은 올바른 의학적 판단범위 내에서 처치의에 의해 결정될 수 있으며, 일반적으로 0.001 내지 1000 mg/kg의 양, 바람직하게는 0.05 내지 200 mg/kg, 보다 바람직하게는 0.1 내지 100 mg/kg의 양을 일일 1회 내지 수회로 나누어 투여할 수 있다.The term "administering" in the present invention means introducing the pharmaceutical composition of the present invention to a suspect subject by any appropriate method, and the administration route is administered through various routes of oral or parenteral administration as long as it can reach the target tissue . The method of treatment of the present invention may comprise administering a pharmaceutically effective amount of the pharmaceutical composition comprising the puritone. Suitable total daily doses may be determined by the treatment within the scope of sound medical judgment and are generally in the range of 0.001 to 1000 mg / kg, preferably 0.05 to 200 mg / kg, more preferably 0.1 to 100 mg / kg can be administered once or several times a day.
그러나 본 발명의 목적상, 특정 환자에 대한 구체적인 치료적 유효량은 달성하고자 하는 반응의 종류와 정도, 경우에 따라 다른 제제가 사용되는지의 여부를 비롯한 구체적 조성물, 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 분비율, 치료기간, 구체적 조성물과 함께 사용되거나 동시 사용되는 약물을 비롯한 다양한 인자와 의약 분야에 잘 알려진 유사 인자에 따라 다르게 적용하는 것이 바람직하다.For purposes of the present invention, however, the specific therapeutically effective amount for a particular patient will depend upon the nature and extent of the reaction to be achieved, the particular composition, including whether or not other agents are used, the age, weight, Sex and diet of the patient, the time of administration, the route of administration and the rate of administration of the composition, the duration of the treatment, the drugs used or concurrently used with the specific composition, and similar factors well known in the medical arts.
한편, 본 발명의 퓨리톤은 천연물질로서 인체에 무해하며, 독성 및 부작용이 거의 없으므로 장기간 사용시에도 안심하고 사용할 수 있으며, 특히 상기한 바와 같은 약학적 및 식품 조성물에 안전하게 적용할 수 있다.On the other hand, the puritone of the present invention is a natural substance which is harmless to the human body and has little toxicity and side effects, so that it can be safely used even in long-term use, and can be safely applied to the pharmaceutical and food composition as described above.
하나의 구체적 양태로, 본 발명은 퓨리톤을 유효성분으로 포함하는 항암용 조성물을 개체에 투여하여 개체의 암을 예방 또는 개선하는 방법 및 그 용도를 제공한다.In one specific embodiment, the present invention provides a method for preventing or improving cancer of an individual by administering to a subject an anti-cancer composition comprising furitone as an active ingredient, and uses thereof.
이와 같이, 본 발명의 퓨리톤은 인간 유방암 세포주 MDA-MB-231, 인간 간암 세포주 HepG2, 인간 폐암 세포주 A549, 인간 난소암 세포주 OVCA 429, 인간 혈액암 세포주 HL60, 인간 피부암 세포주 A375, 인간 신장암 세포주 RCC4(-) 및 인간 대장암 세포주 HCT116의 암세포 증식을 억제할 수 있으므로 항암용 조성물로서 유용하게 사용할 수 있다. 또한 본 발명의 퓨리톤은 천연물질로서 인체에 안전하여 약학적 및 식품 조성물에 안전하게 사용할 수 있다. Thus, the puritone of the present invention can be used as a human breast cancer cell line MDA-MB-231, human liver cancer cell line HepG2, human lung cancer cell line A549, human ovarian cancer
도 1은 Raw264.7 세포에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 세포 생존율을 나타낸 그래프이다.
도 2는 Raw264.7 세포에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양한 세포 사진이다.
도 3은 Raw264.7 세포에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 세포 생존율을 나타낸 그래프이다.
도 4는 Raw264.7 세포에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양한 세포 사진이다.
도 5는 인간 유방암 세포주 MDA-MB-231에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 6은 인간 유방암 세포주 MDA-MB-231에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양한 세포 사진이다.
도 7은 인간 유방암 세포주 MDA-MB-231에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 8은 인간 유방암 세포주 MDA-MB-231에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양한 세포 사진이다.
도 9는 인간 간암 세포주 HepG2에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 10은 인간 간암 세포주 HepG2에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 11은 인간 폐암 세포주 A549에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 12는 인간 폐암 세포주 A549에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 13은 인간 난소암 세포주 OVCA 429에 퓨리톤을 처리한 후 3일 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 14는 인간 난소암 세포주 OVCA 429에 퓨리톤을 농도별로 처리한 후 3일 동안 배양한 세포 사진이다.
도 15는 인간 난소암 세포주 OVCA 429에 100%의 퓨리톤을 처리한 후 3일 동안 배양한 결과이다.
도 16은 인간 혈액암 세포주 HL60에 퓨리톤을 처리한 후 2일 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 17은 인간 피부암 세포주 A375에 퓨리톤을 처리한 후 2일 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 18은 인간 피부암 세포주 A375에 퓨리톤을 농도별로 처리한 후 2일 동안 배양한 세포 사진이다.
도 19는 인간 신장암 세포주 RCC4(-)에 퓨리톤을 처리한 후 2일 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 20은 인간 신장암 세포주 RCC4(-)에 퓨리톤을 농도별로 처리한 후 2일 동안 배양한 세포 사진이다.
도 21은 인간 대장암 세포주 HCT116에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다.
도 22는 인간 대장암 세포주 HCT116에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다. FIG. 1 is a graph showing cell viability by culturing Raw264.7 cells for 24 hours after treating puritone by concentration. FIG.
FIG. 2 is a photograph of cells cultured for 24 hours in the Raw264.7 cells treated with puritone by concentration. FIG.
FIG. 3 is a graph showing cell viability by culturing Raw264.7 cells at different concentrations of puritone for 48 hours. FIG.
Fig. 4 is a photograph of cells cultured for 48 hours in the Raw264.7 cells treated with puritone by concentration.
FIG. 5 is a graph showing the effect of suppressing proliferation of cancer cells by culturing the human breast cancer cell line MDA-MB-231 for 24 hours after treatment with puritone by concentration.
6 is a photograph of cells cultured for 24 hours in a human breast cancer cell line MDA-MB-231 treated with puritone by concentration.
FIG. 7 is a graph showing the effect of suppressing proliferation of cancer cells by culturing the human breast cancer cell line MDA-MB-231 at various concentrations of puritone for 48 hours.
8 is a photograph of cells cultured for 48 hours in a human breast cancer cell line MDA-MB-231 treated with puritone by concentration.
FIG. 9 is a graph showing the effect of inhibiting proliferation of cancer cells by culturing the human liver cancer cell line HepG2 for 24 hours after treatment with puritone by concentration.
FIG. 10 is a graph showing the effect of suppressing proliferation of cancer cells by culturing the human liver cancer cell line HepG2 at different concentrations of puritone for 48 hours.
FIG. 11 is a graph showing the effect of suppressing the proliferation of cancer cells by culturing the human lung cancer cell line A549 for 24 hours after treatment with puritone by concentration.
FIG. 12 is a graph showing the effect of inhibiting proliferation of cancer cells by culturing the human lung cancer cell line A549 at a concentration of puritone by concentration for 48 hours.
13 is a graph showing the inhibitory effect of cancer cell proliferation by culturing human ovarian cancer
FIG. 14 is a photograph of a cell cultured for 3 days after treatment with puritone at a concentration of human ovarian cancer
Fig. 15 shows the result of culturing the human ovarian cancer
FIG. 16 is a graph showing the effect of inhibiting cancer cell proliferation by culturing human blood cancer cell line HL60 for 2 days after treatment with puritone.
FIG. 17 is a graph showing the effect of inhibiting cancer cell proliferation by culturing human skin cancer cell line A375 for 2 days after treatment with puritone.
Fig. 18 is a photograph of a cell cultured for 2 days after treatment with puritone at a concentration of human skin cancer cell line A375.
19 is a graph showing the effect of inhibiting cancer cell proliferation by culturing human renal cancer cell line RCC4 (-) for 2 days after treatment with puritone.
FIG. 20 is a photograph of a cell cultured for two days after treatment of the human renal cell carcinoma cell line RCC4 (-) with puritone by concentration.
FIG. 21 is a graph showing the effect of suppressing proliferation of cancer cells by culturing the human colon cancer cell line HCT116 for 24 hours after treatment with puritone by concentration.
FIG. 22 is a graph showing the effect of suppressing proliferation of cancer cells by culturing the human colon cancer cell line HCT116 for 48 hours after treatment with puritone by concentration. FIG.
이하, 본 발명의 이해를 돕기 위하여 실시예 등을 들어 상세하게 설명하기로 한다. 그러나, 본 발명에 따른 실시예들은 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 하기 실시예들에 한정되는 것으로 해석되어서는 안 된다. 본 발명의 실시예들은 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다. Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.
<실시예 1> 퓨리톤의 세포독성 측정Example 1 Cytotoxicity Measurement of Puritone
퓨리톤이 RAW264.7 세포에서의 세포생존률에 미치는 영향을 확인하기 위하여 하기와 같은 방법으로 실험하였다(Lee, S.U., Choi, Y.H., Kim, Y.S., Min, Y.K., Rhee, M., Kim, S.H., 2010. Anti-resorptive sa,urolactam exhibits in vitro anti-inflammatory activity via ERK-NF-kappaB signaling pathway. International immunopharmacology 10, 298-303.)In order to confirm the effect of puritone on the cell survival rate in RAW264.7 cells, the following experiment was performed (Lee, SU, Choi, YH, Kim, YS, Min, YK, Rhee, , 2010. Anti-resorptive SA, urolactam exhibits in vitro anti-inflammatory activity via ERK-NF-kappaB signaling pathway.
생쥐의 대식세포인 Raw264.7 세포(TIB-71, ATCC)를 소태아혈청(Fetal Bovine Serum)을 10% 및 페니실린 1%를 첨가한 DMEM 배지(Dulbecco's Modified Eagle Medium, Gibco)에 1x104/㎖의 농도로 현탁하여 200 ㎕씩 96 웰 플레이트에 접종하여 12시간동안 부착하였다. 퓨리톤을 농도별(0, 1, 2, 5, 10, 50, 100, 150 ㎕/200 ㎕ (0 - 75%))로 처리한 후, 24시간 및 48시간 동안 배양하였다. Of macrophages in mice Raw264.7 cells (TIB-71, ATCC) of fetal bovine serum (Fetal Bovine Serum) and 10% penicillin and 1% in DMEM culture medium (Dulbecco's Modified Eagle Medium, Gibco ) was added to 1x10 4 / ㎖ , And 200 [mu] L of the solution was inoculated in a 96-well plate and adhered for 12 hours. Puritone was treated with concentration (0, 1, 2, 5, 10, 50, 100, 150 ㎕ / 200 ㎕ (0 - 75%)) and cultured for 24 hours and 48 hours.
이후 10㎕의 MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphinyltetrazolium bromide) 용액 (저장 농도: 5 mg/㎖)을 첨가하고 3시간 동안 추가반응을 유도하였다. 반응 종료 및 포마잔 결정을 용해하기 위하여, 각 웰에 100 ㎕의 MTT 정지 용액 (0.01M HCl 내 10% 소듐 도데실 설페이트)을 추가적으로 첨가하였다. 세포 생존율은 MTT가 포마잔으로 환원된 양을 570 nm에서 흡광도를 분광광도계(Molecular Device, CA, USA)를 이용하여 측정하여 얻어진 OD 값을 통해 산출하였다.Then, 10 μl of MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphinyltetrazolium bromide) solution (storage concentration: 5 mg / ml) was added and further reaction was induced for 3 hours. To terminate the reaction and dissolve the formazan crystals, 100 [mu] l MTT stop solution (10% sodium dodecyl sulfate in 0.01 M HCl) was additionally added to each well. The cell viability was calculated from the OD value obtained by measuring the absorbance at 570 nm using a spectrophotometer (Molecular Device, CA, USA) in which the amount of MTT reduced to formazan was measured.
도 1은 Raw264.7 세포에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 세포 생존율을 나타낸 그래프이고, 도 2는 Raw264.7 세포에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양한 세포 사진이다. 여기에서 보듯이, 세포 배양 24시간 동안 0.5 내지 25%로 세포 증식이 나타났으며, RAW264.7 세포 내에서 세포 생존율의 감소를 거의 나타내지 않아 세포독성이 없음을 확인하였다.FIG. 1 is a graph showing the cell survival rate by culturing Raw264.7 cells in a concentration-dependent manner for 24 hours. FIG. 2 is a graph showing the cell survival rate of Raw264.7 cells treated with puritone It is a photograph. As shown here, cell proliferation was observed at 0.5 to 25% for 24 hours in the cell culture, and there was little decrease in cell survival rate in RAW264.7 cells, and it was confirmed that there was no cytotoxicity.
도 3은 Raw264.7 세포에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 세포 생존율을 나타낸 그래프이고, 도 4는 Raw264.7 세포에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양한 세포 사진이다. 여기에서 보듯이, 세포 배양 48시간 동안 0.5 내지 25%로 세포 증식이 나타났으며, RAW264.7 세포 내에서 세포 생존율의 감소를 거의 나타내지 않아 세포독성이 없음을 확인하였다.FIG. 3 is a graph showing cell viability by culturing Raw264.7 cells in a concentration-dependent manner at a concentration of puritone for 48 hours. FIG. 4 is a graph showing the cell viability of Raw264.7 cells after 48 hours of treatment with puritone It is a photograph. As shown here, cell proliferation was observed at 0.5 to 25% for 48 hours in the cell culture, and there was little decrease in cell survival rate in RAW264.7 cells, confirming no cytotoxicity.
<실시예 2> 퓨리톤의 유방암 세포주 MDA-MB-231에 대한 증식 억제효과 측정Example 2 Measurement of proliferation inhibitory effect of puritone on breast cancer cell line MDA-MB-231
퓨리톤이 유방암 세포주 MDA-MB-231에서의 암세포 증식 억제효과를 확인하기 위하여 하기와 같은 방법으로 실험하였다Puritone was tested in the following manner to confirm the inhibitory effect of cancer cell growth on the breast cancer cell line MDA-MB-231
인간 유방암 세포주 MDA-MB-231를 소태아혈청(Fetal Bovine Serum)을 10% 및 페니실린 1%를 첨가한 DMEM 배지(Dulbecco's Modified Eagle Medium, Gibco)에 1x104/㎖의 농도로 현탁하여 200 ㎕씩 96 웰 플레이트에 접종하여 12시간동안 부착하였다. 퓨리톤을 농도별(0, 1, 2, 5, 10, 50, 100 ㎕/200 ㎕ (0 - 55%))로 처리한 후, 24시간 및 48시간 동안 배양하였다. The human breast cancer cell line MDA-MB-231 was suspended in DMEM medium (Dulbecco's Modified Eagle Medium, Gibco) supplemented with 10% fetal bovine serum and 1% penicillin at a concentration of 1 x 10 4 / Inoculated on a 96-well plate and allowed to attach for 12 hours. Puritone was treated with concentrations (0, 1, 2, 5, 10, 50, 100 μl / 200 μl (0 - 55%)) and cultured for 24 and 48 hours.
이후 10㎕의 MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphinyltetrazolium bromide) 용액 (저장 농도: 5 mg/㎖)을 첨가하고 3시간 동안 추가반응을 유도하였다. 반응 종료 및 포마잔 결정을 용해하기 위하여, 각 웰에 100 ㎕의 MTT 정지 용액 (0.01M HCl 내 10% 소듐 도데실 설페이트)을 추가적으로 첨가하였다. 세포 생존율은 MTT가 포마잔으로 환원된 양을 570 nm에서 흡광도를 분광광도계(Molecular Device, CA, USA)를 이용하여 측정하여 얻어진 OD 값을 통해 산출하였다.Then, 10 μl of MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphinyltetrazolium bromide) solution (storage concentration: 5 mg / ml) was added and further reaction was induced for 3 hours. To terminate the reaction and dissolve the formazan crystals, 100 [mu] l MTT stop solution (10% sodium dodecyl sulfate in 0.01 M HCl) was additionally added to each well. The cell viability was calculated from the OD value obtained by measuring the absorbance at 570 nm using a spectrophotometer (Molecular Device, CA, USA) in which the amount of MTT reduced to formazan was measured.
도 5는 인간 유방암 세포주 MDA-MB-231에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이고, 도 6은 인간 유방암 세포주 MDA-MB-231에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양한 세포 사진이다. 여기에서 보듯이, 세포 배양 24시간 동안 25 내지 50%로 암세포 증식 억제효과를 나타냄을 알 수 있었다.FIG. 5 is a graph showing the effect of inhibiting the proliferation of cancer cells by culturing the human breast cancer cell line MDA-MB-231 for 24 hours after treatment with puritone by concentration. FIG. 6 is a graph showing the effect of purine on the human breast cancer cell line MDA- And then cultured for 24 hours. As shown here, it was found that the cell growth inhibition effect was 25 to 50% for 24 hours.
도 7은 인간 유방암 세포주 MDA-MB-231에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이고, 도 8은 인간 유방암 세포주 MDA-MB-231에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양한 세포 사진이다. 여기에서 보듯이, 세포 배양 동안 25 내지 50%로 암세포 증식 억제효과를 나타냄을 알 수 있었다.FIG. 7 is a graph showing the effect of inhibiting proliferation of cancer cells by culturing the human breast cancer cell line MDA-MB-231 at different concentrations by puritone treatment for 48 hours, and FIG. 8 is a graph showing the effect of suppressing the proliferation of human breast cancer cell line MDA- And then cultured for 48 hours. As shown here, it was found that the cell growth inhibitory effect was 25 to 50% during cell culture.
<실시예 3> 퓨리톤의 간암 세포주 HepG2에 대한 증식 억제효과 측정<Example 3> Measurement of proliferation inhibitory effect on puritanone hepatocellular cell line HepG2
퓨리톤이 간암 세포주 HepG2에서의 암세포 증식 억제효과를 확인하기 위하여 하기와 같은 방법으로 실험하였다.Puritone was tested by the following method in order to confirm the inhibitory effect of cancer cell growth on liver cancer cell line HepG2.
인간 간암 세포주 HepG2를 소태아혈청(Fetal Bovine Serum)을 10% 및 페니실린 1%를 첨가한 DMEM 배지(Dulbecco's Modified Eagle Medium, Gibco)에 1x104/㎖의 농도로 현탁하여 200 ㎕씩 96 웰 플레이트에 접종하여 12시간동안 부착하였다. 퓨리톤을 농도별(0, 1, 2, 5, 10, 50, 100, 150 ㎕/200 ㎕ (0 - 75%))로 처리한 후, 24시간 및 48시간 동안 배양하였다. Human liver cancer cell line HepG2 was suspended in DMEM medium (Dulbecco's Modified Eagle Medium, Gibco) supplemented with 10% fetal bovine serum and 1% penicillin at a concentration of 1 x 10 4 / ml, And inoculated for 12 hours. Puritone was treated with concentration (0, 1, 2, 5, 10, 50, 100, 150 ㎕ / 200 ㎕ (0 - 75%)) and cultured for 24 hours and 48 hours.
이후 10㎕의 MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphinyltetrazolium bromide) 용액 (저장 농도: 5 mg/㎖)을 첨가하고 3시간 동안 추가반응을 유도하였다. 반응 종료 및 포마잔 결정을 용해하기 위하여, 각 웰에 100 ㎕의 MTT 정지 용액 (0.01M HCl 내 10% 소듐 도데실 설페이트)을 추가적으로 첨가하였다. 세포 생존율은 MTT가 포마잔으로 환원된 양을 570 nm에서 흡광도를 분광광도계(Molecular Device, CA, USA)를 이용하여 측정하여 얻어진 OD 값을 통해 산출하였다.Then, 10 μl of MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphinyltetrazolium bromide) solution (storage concentration: 5 mg / ml) was added and further reaction was induced for 3 hours. To terminate the reaction and dissolve the formazan crystals, 100 [mu] l MTT stop solution (10% sodium dodecyl sulfate in 0.01 M HCl) was additionally added to each well. The cell viability was calculated from the OD value obtained by measuring the absorbance at 570 nm using a spectrophotometer (Molecular Device, CA, USA) in which the amount of MTT reduced to formazan was measured.
도 9는 인간 간암 세포주 HepG2에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다. 여기에서 보듯이, 세포 배양 24시간 동안 50 내지 70%로 암세포 증식 억제효과를 나타냄을 알 수 있었다.FIG. 9 is a graph showing the effect of inhibiting proliferation of cancer cells by culturing the human liver cancer cell line HepG2 for 24 hours after treatment with puritone by concentration. As shown here, it was found that the cell growth inhibition effect was 50 to 70% for 24 hours.
도 10은 인간 간암 세포주 HepG2에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다. 여기에서 보듯이, 세포 배양 48시간 동안 50 내지 75%로 암세포 증식 억제효과를 나타냄을 알 수 있었다.FIG. 10 is a graph showing the effect of suppressing proliferation of cancer cells by culturing the human liver cancer cell line HepG2 at different concentrations of puritone for 48 hours. As shown here, it was found that the cell growth inhibition effect was 50 to 75% for 48 hours.
<실시예 4> 퓨리톤의 폐암 세포주 A549에 대한 증식 억제효과 측정Example 4 Measurement of proliferation inhibitory effect on puritanone lung cancer cell line A549
퓨리톤이 폐암 세포주 A549에서의 암세포 증식 억제효과를 확인하기 위하여 하기와 같은 방법으로 실험하였다.Puritone was tested by the following method to confirm the inhibitory effect of cancer cell line A549 on cancer cell proliferation.
인간 폐암 세포주 A549를 소태아혈청(Fetal Bovine Serum)을 10% 및 페니실린 1%를 첨가한 RPMI 1640 배지에 1x104/㎖의 농도로 현탁하여 200 ㎕씩 96 웰 플레이트에 접종하여 12시간동안 부착하였다. 퓨리톤을 농도별(0, 1, 2, 5, 10, 50, 100, 150 ㎕/200 ㎕ (0 - 75%))로 처리한 후, 24시간 및 48시간 동안 배양하였다. Human lung cancer cell line A549 was suspended in RPMI 1640 medium supplemented with 10% fetal bovine serum (Fetal Bovine Serum) and 1% penicillin at a concentration of 1x10 4 / ml, and 200 μl of each was inoculated in a 96-well plate and adhered for 12 hours . Puritone was treated with concentration (0, 1, 2, 5, 10, 50, 100, 150 ㎕ / 200 ㎕ (0 - 75%)) and cultured for 24 hours and 48 hours.
이후 10㎕의 MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphinyltetrazolium bromide) 용액 (저장 농도: 5 mg/㎖)을 첨가하고 3시간 동안 추가반응을 유도하였다. 반응 종료 및 포마잔 결정을 용해하기 위하여, 각 웰에 100 ㎕의 MTT 정지 용액 (0.01M HCl 내 10% 소듐 도데실 설페이트)을 추가적으로 첨가하였다. 세포 생존율은 MTT가 포마잔으로 환원된 양을 570 nm에서 흡광도를 분광광도계(Molecular Device, CA, USA)를 이용하여 측정하여 얻어진 OD 값을 통해 산출하였다.Then, 10 μl of MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphinyltetrazolium bromide) solution (storage concentration: 5 mg / ml) was added and further reaction was induced for 3 hours. To terminate the reaction and dissolve the formazan crystals, 100 [mu] l MTT stop solution (10% sodium dodecyl sulfate in 0.01 M HCl) was additionally added to each well. The cell viability was calculated from the OD value obtained by measuring the absorbance at 570 nm using a spectrophotometer (Molecular Device, CA, USA) in which the amount of MTT reduced to formazan was measured.
도 11은 인간 폐암 세포주 A549에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다. 여기에서 보듯이, 세포 배양 24시간 동안 50 내지 70%로 암세포 증식 억제효과를 나타냄을 알 수 있었다.FIG. 11 is a graph showing the effect of suppressing the proliferation of cancer cells by culturing the human lung cancer cell line A549 for 24 hours after treatment with puritone by concentration. As shown here, it was found that the cell growth inhibition effect was 50 to 70% for 24 hours.
도 12는 인간 폐암 세포주 A549에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다. 여기에서 보듯이, 세포 배양 48시간 동안 50 내지 75%로 암세포 증식 억제효과를 나타냄을 알 수 있었다.FIG. 12 is a graph showing the effect of inhibiting proliferation of cancer cells by culturing the human lung cancer cell line A549 at a concentration of puritone by concentration for 48 hours. As shown here, it was found that the cell growth inhibition effect was 50 to 75% for 48 hours.
<실시예 5> 퓨리톤의 난소암 세포주 OVCA 429에 대한 증식 억제효과 측정Example 5 Measurement of proliferation inhibitory effect on puritan ovarian cancer
퓨리톤이 난소암 세포주(Ovary cancer cell) OVCA 249에서의 암세포 증식 억제효과를 하기와 같은 방법으로 실험하였다.Puritone inhibited cancer cell proliferation in ovarian cancer cell OVCA 249 by the following method.
계대 배양 24 시간 후, 96-웰 플레이트 내의 세포를 퓨리톤 및 H2O로 처리한 다음 pH12로 조절하여 1 일 동안 처리하였다. 세포 생존력은 CellTiter-Glo 발광 세포 생존력 분석법 (Promega Corporation, Madison, WI, USA)을 사용하여 측정하였다. 96-웰 플레이트를 약 30분 동안 실온으로 간단히 평형시켰다. CellTiter-Glo 시약 (100 μl)을 각 웰에 넣었다. 이어서, Tecan Infinite F200 마이크로 플레이트 판독기를 사용하여 배지 및 시약을 오비탈 쉐이커(orbital shaker)에서 2분 동안 혼합하고 실온에서 10 분 동안 방치하여 발광을 기록하였다. 발광 최종 값은 상대 백분율로 나타내었다.After 24 hours of passage, cells in 96-well plates were treated with puritone and H2O and then adjusted to pH 12 for 1 day. Cell viability was measured using CellTiter-Glo luminescence cell viability assay (Promega Corporation, Madison, WI, USA). The 96-well plate was briefly equilibrated to room temperature for about 30 minutes. CellTiter-Glo reagent (100 [mu] l) was added to each well. The medium and reagents were then mixed in an orbital shaker for 2 minutes using a Tecan Infinite F200 microplate reader and allowed to stand at room temperature for 10 minutes to record the luminescence. The final luminosity was expressed as a relative percentage.
도 13은 인간 난소암 세포주 OVCA 429에 퓨리톤을 처리한 후 3일 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이고, 도 14는 인간 난소암 세포주 OVCA 429에 퓨리톤을 농도별로 처리한 후 3일 동안 배양한 세포 사진이다. 여기에서 보듯이, 70%의 농도로 처리된 퓨리톤은 99 % OVCA429 세포 성장 억제효능을 나타내었다.FIG. 13 is a graph showing inhibitory effect on cancer cell proliferation by culturing human ovarian cancer
도 15는 인간 난소암 세포주 OVCA 429에 100%의 퓨리톤을 처리한 후 3일 동안 배양한 결과이다. 여기에서 보듯이, 퓨리톤은 99 % OVCA429 세포 성장 억제효능을 나타내었다.Fig. 15 shows the result of culturing the human ovarian cancer
<실시예 6> 퓨리톤의 혈액암 세포주 HL60에 대한 증식 억제효과 측정Example 6 Measurement of Proliferation Inhibitory Effect of Puritone on HL60 Blood Cancer Cell Line
퓨리톤이 혈액암 세포주 (Promyelocytic Leukemia) HL60에서의 암세포 증식 억제효과를 상기 실시예 5와 같은 방법으로 실험하였다.Puritone inhibited cancer cell proliferation in Promyelocytic Leukemia HL60 in the same manner as in Example 5 above.
도 16은 인간 혈액암 세포주 HL60에 퓨리톤을 처리한 후 2일 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이이다. 여기에서 보듯이, 50 % 퓨리톤 처리시 HL60 세포 성장 억제가 70 %로 나타났으며, 70% 퓨리톤 처리시 99% 의 HL60 세포 성장 억제효과를 나타내었다. 16 is a graph showing the inhibitory effect of cancer cell proliferation by culturing human blood cancer cell line HL60 for 2 days after treatment with puritone. As shown here, HL60 cell growth inhibition was 70% at 50% puritone treatment and 99% HL60 cell growth inhibition at 70% puritone treatment.
<실시예 7> 퓨리톤의 피부암 세포주 A375에 대한 증식 억제효과 측정Example 7 Measurement of Proliferation Inhibitory Effect on Puritone Skin Cancer Cell Line A375
퓨리톤이 피부암 세포주 (Melanoma Cell) A375에서의 암세포 증식 억제효과를 상기실시예 5와 같은 방법으로 실험하였다.Puritone inhibited cancer cell proliferation in melanoma cell A375 in the same manner as in Example 5 above.
도 17은 인간 피부암 세포주 A375에 퓨리톤을 처리한 후 2일 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이고, 도 19는 인간 피부암 세포주 A375에 퓨리톤을 농도별로 처리한 후 2일 동안 배양한 세포 사진이다. 여기에서 보듯이, 50 % 퓨리톤 처리시 A375 세포 성장 억제가 20 %로 나타났으며, 70% 퓨리톤 처리시 99% 의 HL60 세포 성장 억제효과를 나타내었다. FIG. 17 is a graph showing the inhibitory effect of cancer cell proliferation inhibition by culturing human skin cancer cell line A375 for 2 days after treatment with puritone. FIG. 19 is a graph showing the effect of puritone treatment on human skin cancer cell line A375 It is a photograph. As shown here, A375 cell growth inhibition was 20% at 50% puritone treatment and 99% HL60 cell growth inhibition at 70% puritone treatment.
<실시예 8> 퓨리톤의 신장암 세포주 RCC4(-)에 대한 증식 억제효과 측정Example 8 Measurement of Proliferation Inhibitory Effect of Puritone on Renal Cancer Cell Line RCC4 (-)
퓨리톤이 신장암 세포주(Ranal cancer cell) RCC4(-)에서의 암세포 증식 억제효과를 상기와 같은 방법으로 실험하였다.Puritone inhibited the cancer cell proliferation inhibition in the renal cancer cell RCC4 (-) as described above.
도 19는 인간 신장암 세포주 RCC4(-)에 퓨리톤을 처리한 후 2일 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이고, 도 20은 인간 신장암 세포주 RCC4(-)에 퓨리톤을 농도별로 처리한 후 2일 동안 배양한 세포 사진이다. 여기에서 보듯이, 70 % 퓨리톤 처리시 RCC4(-) 세포 성장 억제가 99 %로 나타났다. FIG. 19 is a graph showing the cancer cell proliferation inhibitory effect by culturing the human renal cell carcinoma cell line RCC4 (-) for 2 days after treatment with puritone. FIG. 20 is a graph showing the effect of treating the human renal cancer cell line RCC4 And then cultured for 2 days. As shown here, the inhibition of RCC4 (-) cell growth by 70% puritone treatment was 99%.
<실시예 9> 퓨리톤의 대장암 세포주 HCT116에 대한 증식 억제효과 측정Example 9 Measurement of Proliferation Inhibitory Effect of Puritone on Colon Cancer Cell Line HCT116
퓨리톤이 대장암 세포주(colorectal cancer cell line) HCT116에서의 암세포 증식 억제효과를 상기와 같은 방법으로 실험하였다.Puritone inhibited cancer cell proliferation in colorectal cancer cell line HCT116 in the same manner as described above.
도 21은 인간 대장암 세포주 HCT116에 퓨리톤을 농도별로 처리한 후 24시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다. 여기에서 보듯이, 세포 배양 24시간 동안 50 내지 90%로 암세포 증식 억제효과를 나타냄을 알 수 있었다.FIG. 21 is a graph showing the effect of suppressing proliferation of cancer cells by culturing the human colon cancer cell line HCT116 for 24 hours after treatment with puritone by concentration. As shown here, the inhibitory effect on cancer cell proliferation was shown to be 50 to 90% for 24 hours of cell culture.
도 22는 인간 대장암 세포주 HCT116에 퓨리톤을 농도별로 처리한 후 48시간 동안 배양하여 암세포 증식 억제효과를 나타낸 그래프이다. 여기에서 보듯이, 세포 배양 48시간 동안 50 내지 60%로 암세포 증식 억제효과를 나타냄을 알 수 있었다.FIG. 22 is a graph showing the effect of suppressing proliferation of cancer cells by culturing the human colon cancer cell line HCT116 for 48 hours after treatment with puritone by concentration. FIG. As shown here, it was found that the cell growth inhibition effect was 50 to 60% for 48 hours.
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| PCT/KR2018/015205 WO2019108049A2 (en) | 2017-12-01 | 2018-12-03 | Anticancer, antibacterial, alcohol-degrading, skin wound-healing, skin-regenerating, or antiasthmatic composition comprising mineral ion mixture |
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| KR1020170164585 | 2017-12-01 | ||
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| KR1020190068097A Division KR102250733B1 (en) | 2017-12-01 | 2019-06-10 | An anticancer composition comprising purtiton |
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| KR100452007B1 (en) | 2003-05-07 | 2004-10-08 | 박용진 | Mineral powder having inhibition effect of cancer cell proliferation |
| DE10323759A1 (en) | 2003-05-22 | 2004-12-16 | Bauer, Wulf, Dr. | Remedies for internal use, especially against cancer |
| KR100454200B1 (en) | 2003-09-03 | 2004-10-27 | (주)서봉바이오베스텍 | Germanium mixed mineral having anti-cancer activating ingredient and its use as anti-cancer medicine |
| KR101268434B1 (en) | 2010-04-26 | 2013-06-04 | 이종두 | Anticancer agent having immunomodulatory function comprising silicate mineral as an effective component and manufacturing method thereof |
| KR101199897B1 (en) | 2011-11-14 | 2012-11-09 | 권태동 | Pharmaceutical compositions for prevention and treatment of cancer comprising natural mineral and lysate thereof as an active ingredient |
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Non-Patent Citations (1)
| Title |
|---|
| 인터넷자료. Experiment Result of Puriton/Cell Viability Assay. http://www.puriton.kr/article/%EC%97%B0%EA%B5%AC%EA%B2%B0%EA%B3%BC-%EC%9E%90%EB%A3%8C%EC%8B%A4/7/12/ (2017.6.23.)* |
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| KR102250733B1 (en) | 2021-05-11 |
| KR20190068511A (en) | 2019-06-18 |
| KR20190067077A (en) | 2019-06-14 |
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