KR101394292B1 - A benzoflavonol derivative, preparing method of the same and use in anti-cancer agent thereof - Google Patents

A benzoflavonol derivative, preparing method of the same and use in anti-cancer agent thereof Download PDF

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KR101394292B1
KR101394292B1 KR1020120117404A KR20120117404A KR101394292B1 KR 101394292 B1 KR101394292 B1 KR 101394292B1 KR 1020120117404 A KR1020120117404 A KR 1020120117404A KR 20120117404 A KR20120117404 A KR 20120117404A KR 101394292 B1 KR101394292 B1 KR 101394292B1
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benzoflavone
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임융호
이영한
고동수
신순영
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건국대학교 산학협력단
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Abstract

본 발명은 벤조플라보놀 유도체, 그 제법 및 항암제로서의 용도에 관한 것으로, 더욱 상세하게는 벤조플라보놀 유도체의 사람 대장암 세포에서 세포사멸 유도 효과를 갖는 항암제용 약학조성물에 관한 것이다.The present invention relates to a benzoflavone derivative, its preparation and its use as an anticancer agent, and more particularly, to a pharmaceutical composition for an anticancer agent having an effect of inducing apoptosis in human colorectal cancer cells of a benzoflavone derivative.

Description

벤조플라보놀 유도체 및 그 제법 및 항암제로서의 용도 {A benzoflavonol derivative, preparing method of the same and use in anti-cancer agent thereof}Benzoflavonol derivatives, preparation thereof, and use thereof as anticancer agents.

본 발명은 벤조플라보놀 유도체, 그 제법 및 항암제로서의 용도에 관한 것으로, 더욱 상세하게는 벤조플라보놀 유도체의 사람 대장암 세포에서 세포사멸 유도 효과를 갖는 항암제용 약학조성물에 관한 것이다.
The present invention relates to a benzoflavone derivative, its preparation and its use as an anticancer agent, and more particularly, to a pharmaceutical composition for an anticancer agent having an effect of inducing apoptosis in human colorectal cancer cells of a benzoflavone derivative.

세포가 사멸하는 방법은 외부의 물리적 자극에 의해 사멸되는 괴사(Necrosis)와 세포 신호전달계의 활성 또는 비활성 조절 기작에 의해 계획된 사멸 과정이 유도되는 세포자기사멸(Programmed cell death)로 구분할 수 있다. 괴사에 의한 사멸은 세포 자체가 팽창하고 파열되는 반응으로 손상된 세포 주변에는 염증이 유발되기도 한다. 세포자기사멸은 불완전한 세포의 성장을 제어하기 위한 과정으로 세포의 정상적인 기능과 양적 균형을 이루기위해서 스스로 조절하는 반응이다. 세포자기사멸은 type-I (apoptosis)과 type-II (autophagic cell death) 두가지 형태가 있다. 일반적으로 type-I 자기사멸 현상은 염색사(Chromatin)의 응축 (Condensation)과 DNA의 단편 형성 그리고 작은 세포 조각(Apoptotic body)들이 생성되는 과정으로 진행된다. 세포자기사멸에 의한 과정은 주변 세포의 탐식 작용(phagocytosis)으로 염증을 유발 하지 않는 것이 특징이다 (Experimental Cell Research 256,12-18,2000).The way in which the cells die is divided into necrosis, which is killed by external physical stimulation, and programmed cell death, in which a planned death process is induced by the activation or inactivation of the cell signal transduction system. Necrotic death is a reaction in which the cell itself expands and ruptures, causing inflammation around the damaged cells. Cell self-apoptosis is a process for controlling the growth of incomplete cells and is a self-regulating response to quantitative balance with the normal function of cells. There are two types of apoptosis: type-I (apoptosis) and type-II (autophagic cell death). In general, type-I self-destruction proceeds to the process of condensation of chromatin, fragmentation of DNA, and generation of apoptotic bodies. The process by apoptosis is characterized by the phagocytosis of surrounding cells, which does not cause inflammation (Experimental Cell Research 256, 12-18, 2000).

카스파제 (caspase)는 세포 사멸을 유도시키는 씨스테인 단백분해 효소 (cystein protease)중의 하나로서, 통상 비활성형의 전구효소(proenzyme)인 프로카스파제(procaspase) 형태로 존재하고 있다. 프로카스파제 단백질은 pro-domain과 small-subunit, large-subunit 등의 구조로 구성되어있는데, 세포사멸 자극 신호에 의해 pro-domain과 large-subunit 사이 와 small-subunit 와 large-subunit 사이가 잘려진 후 small-subunit와 large-subunit 끼리만 다시 결합하여 활성형 효소로 전환된다. DNA 손상 자극에 의한 세포사멸 현상 과정의 가장 대표적인 작용 기전은 세포내로 전달된 각종 DNA 손상 자극에 의해 p53 유전자가 활성되면서 시작된다. 활성된 p53 유전자는 Bax나 PUMA, NOXA 같은 단백질 발현을 증가시켜 미토콘드리아 막에 존재하는 씨토크롬-C (cytochrome c) 단백질을 유리시키고, 유리된 씨토크롬-C 단백질은 Apaf-1 단백질과 카스파제-9 전구효소 (procaspase-9)와 결합하여 아포프토좀 (apoptosome)이라는 단백 복합체를 이루어 카스파제-9 효소를 활성시킨다 (Cell 1997,91,479-489; Mol Cell 1998,1,949-957; Cell 1998,94,739-750). 활성 카스파제-9은 곧 이어서 비활성 카스파제-3 또는 카스파제-7 전구효소(proenzyme)를 절단시켜 이들 효소를 활성시킨다 (Cell 1998,94,339-352). 활성형 카스파제-3 또는 카스파제-7은 세포의 생리적 활성에 중요한 각종 기질 단백질을 분해시켜 결과적으로 세포의 죽음이 유도된다. The caspase is one of the cystein proteases that induce apoptosis, and is present in the form of a procaspase, which is usually an inactive proenzyme. Pro-caspase proteins consist of pro-domain, small-subunit, and large-subunit structures. The pro-domain and large-subunit and the small-subunit and large-subunit are cleaved Small-subunits and large-subunits are recombined and converted to active enzymes. The most prominent mechanism of apoptosis by DNA damage stimulation begins with the activation of the p53 gene by various DNA damage stimuli delivered into the cell. The activated p53 gene increases protein expression such as Bax, PUMA, and NOXA to liberate the cytochrome c protein present in the mitochondrial membrane, and the liberated cytochrome-C protein binds to the Apaf-1 protein and caspase- 9 enzyme by activating caspase-9 enzyme by forming a protein complex called apoptosome in association with procaspase-9 (Cell 1997, 91, 479-489; Mol Cell 1998, 949-957; Cell 1998,94,739 -750). Activated caspase-9 immediately activates these enzymes by cleaving the inactive caspase-3 or caspase-7 proenzyme (Cell 1998, 94, 339-352). The active caspase-3 or caspase-7 degrades various substrate proteins important for the physiological activity of the cells, resulting in cell death.

카스파제-7은 다른 종류의 카스파제와 유사하게 비활성형의 전구체(proenzyme)로 존재하다가 세포사멸 신호 자극에 의해 카스파제-9에 의해 잘려져서 활성형으로 변하게 된다. 카스파제-7은 세포사멸 과정의 마지막 단계에서 활성되면서 세포 활성에 중요한 여러 가지 기질 단백질을 절단함으로써 세포사멸을 유도시킨다. PARP(Poly(ADP-Ribose) Polymerase) 단백질은 손상된 DNA를 복구시켜주는 113 kDa 크기의 DNA 수복 효소이다. PARP 단백질은 type-I 세포자기사멸 과정 중 활성형의 카스파제-3 또는 카스파제-7 효소에 의해 89-kDa과 24-kDa 크기의 단편으로 잘려지기 때문에, PARP 단백질의 89-kDa과 24-kDa 단편의 생성 현상은 type-I 세포 자기사멸 과정을 검증하는 주요 생화학적 표지로 간주 되고 있다 (Nature (1994) 371:346-347). 암세포는 활발하게 분화하므로 세포사멸에 의한 영향이 정상세포보다 훨씬 크게 되므로 세포사멸을 유도함으로써 암세포를 죽이고자 하는 시도가 항암제 개발에서 활발하게 연구되고 있다. Caspase-7 exists as an inactive precursor (proenzyme) similar to other types of caspases, and is cleaved by caspase-9 by cell death signaling to become active. Caspase-7 is activated at the end of the cell death process and induces apoptosis by cleaving several substrate proteins important for cell activation. The PARP (Poly (ADP-Ribose) Polymerase) protein is a 113 kDa DNA repair enzyme that restores damaged DNA. Since the PARP protein is cleaved into 89-kDa and 24-kDa fragments by the active caspase-3 or caspase-7 enzyme during type I cell apoptosis, the 89-kDa and 24- kDa fragment is considered to be a major biochemical marker for verifying the type-I cell apoptotic process (Nature (1994) 371: 346-347). Since cancer cells actively differentiate, the effect of apoptosis is much greater than that of normal cells, and thus attempts to kill cancer cells by inducing apoptosis have been actively studied in the development of anticancer drugs.

과거와는 달리 현재는 낮은 독성을 갖는 물질을 항암제로 찾고자하는 노력이 활발하다. 식물은 동물과 달리 고정상태로 살아가기 때문에 자신의 방어를 위해서 이차대사물을 극미량 생산하는데 알칼로이드, 터르피노이드, 폴리페놀 등이 대표적인 이차대사물에 속한다. 폴리페놀은 다른 이차대사물과는 달리 생물학적 효능이 낮아서 과거에는 항암제로의 개발 대상에서 제외되어 왔으나 근래에는 폴리페놀이 갖는 낮은 독성에 관심을 갖게되어 폴리페놀도 항암제로 개발하고자 하는 노력이 활발하다. 폴피페놀을 탄소 골격에 따라 여러 종류의 패밀리로 구분되는데 이들 중 C6-C3-C6 탄소 골격을 갖는 패밀리를 플라보노이드라고 구분한다. 이와 같은 탄소 골격을 가진 플라보노이드에 벤진환이 하나 더 추가된 물질은 벤조플라보노이드로 구별되는데 식물에서는 일반적으로 알파벤조플라본이 발견되고 베타벤조플라본은 거의 발견되지 않고 단지 합성으로만 얻을 뿐이다. 플라보노이드는 폴리페놀의 일종이므로 히드록시기 또는 메톡시기와 같은 다양한 치환기들의 첨가로 인해 다양한 유도체가 만들어질 수 있다. Unlike in the past, efforts to search for substances with low toxicity as anticancer drugs are now active. Unlike animals, plants live in a fixed state, so for their defense to produce secondary metabolites in a very small amount of alkaloids, terpinoids, polyphenols are among the representative secondary metabolites. Polyphenol, unlike other secondary metabolites, has a low biological efficacy and has previously been excluded from development as an anticancer drug. However, since it has recently been interested in the low toxicity of polyphenols, efforts to develop polyphenol as an anticancer drug have been actively conducted . Folpiphenol is divided into several families depending on the carbon skeleton. Of these, a family having a C6-C3-C6 carbon skeleton is called a flavonoid. Benzene flavonoids are distinguished from flavonoids with carbon skeletons by the addition of benzyne flavonoids. In plants, alpha-benzoflavones are generally found and beta-benzoflavones are rarely found and only synthesized. Because flavonoids are a kind of polyphenols, various derivatives can be made by the addition of various substituents such as hydroxy groups or methoxy groups.

관련 선행특허로 대한민국 특허출원번호 제10-2005-0038628호는 와송의 열수추출물 유래 항암 활성 다당체 또는 올리고당류를 유효성분으로 포함하는 대장암 치료용 조성물에 관한 것으로, 와송의 열수추출물로부터 다당체 및 올리고당류를 추출한 다음 상기 와송 유래 다당체 및 올리고당류가 대장암 세포의 형태, 대장암 세포 성장 억제, 아폽토시스, 대장암 세포의 주기 및 유전자 발현에 미치는 영향을 조사함으로써 와송의 열수추출물 유래 다당체 또는 올리고당류가 대장암에 대하여 항암 활성이 있음을 확인함으로써 와송의 열수추출물 유래 항암 활성 다당체 또는 올리고당류를 유효성분으로 포함하는 대장암 치료용 조성물을 제공할 수 있는 매우 뛰어난 효과가 있다고 기재되어 있다.Korean Patent Application No. 10-2005-0038628 discloses a composition for the treatment of colorectal cancer comprising an anticancer active polysaccharide or oligosaccharide derived from a hot-water extract of Watson as an active ingredient, wherein the polysaccharide and oligosaccharide Derived polysaccharides and oligosaccharides on the morphology of colon cancer cells, inhibition of colon cancer cell growth, apoptosis, cycle of colon cancer cells and gene expression were investigated. As a result, polysaccharides or oligosaccharides derived from the hot- It has been reported that it is possible to provide a composition for the treatment of colorectal cancer comprising, as an active ingredient, an anticancer active polysaccharide or oligosaccharide derived from a hot-water extract of Tohsang by confirming anticancer activity against colon cancer.

본 발명은 상기의 필요성에 의하여 고안된 것으로서 본 발명의 목적은 신규한 항암제를 제공하는 것이다. The present invention has been devised in view of the above needs, and an object of the present invention is to provide a novel anticancer agent.

본 발명의 다른 목적은 신규한 항암제 제조방법을 제공하는 것이다.Another object of the present invention is to provide a novel method for producing an anticancer agent.

본 발명의 또 다른 목적은 대장암 예방 및 치료용 조성물을 제공하는 것이다.It is still another object of the present invention to provide a composition for the prevention and treatment of colorectal cancer.

상기 목적을 달성하기 위하여, 본 발명은 하기 화학식 1의 화합물을 제공한다.In order to achieve the above object, the present invention provides a compound represented by the following general formula (1).

Figure 112012085906612-pat00001
Figure 112012085906612-pat00001

[화학식 1][Chemical Formula 1]

본 발명의 일 구현예에 있어서, 상기 화합물은 카스파제 효소의 활성화를 유도하는 것이 바람직하고, 폴리(ADP-리보스)폴리머레이즈 단백질의 절단을 유도하는 것이 바람직하나 이에 한정되지 아니한다.In one embodiment of the present invention, the compound preferably induces activation of the caspase enzyme, and preferably induces cleavage of the poly (ADP-ribose) polymerase protein, but is not limited thereto.

또 본 발명은 하기 화학식 1의 화합물을 유효성분으로 포함하는 조성물을 포함한다:The present invention also encompasses a composition comprising, as an active ingredient, a compound of formula (1): < EMI ID =

Figure 112012085906612-pat00002
Figure 112012085906612-pat00002

[화학식 1][Chemical Formula 1]

또 본 발명은 하기 화학식 1의 화합물 및 약학적으로 허용가능한 담체를 유효성분으로 포함하는 약학 조성물을 제공한다:The present invention also provides a pharmaceutical composition comprising, as an active ingredient, a compound represented by the following formula (1) and a pharmaceutically acceptable carrier:

Figure 112012085906612-pat00003
Figure 112012085906612-pat00003

[화학식 1][Chemical Formula 1]

또 본 발명은 하기 화학식 1의 화합물 및 약학적으로 허용가능한 담체를 유효성분으로 포함하는 항암용 약학 조성물을 제공한다:The present invention also provides an anticancer pharmaceutical composition comprising, as an active ingredient, a compound represented by the following formula (1) and a pharmaceutically acceptable carrier:

Figure 112012085906612-pat00004
Figure 112012085906612-pat00004

[화학식 1][Chemical Formula 1]

본 발명의 일 구현예에 있어서, 상기 조성물은 대장암에 대한 항암효과를 가지는 것이 바람직하나 이에 한정되지 아니한다.In one embodiment of the present invention, the composition preferably has anticancer effect on colon cancer, but is not limited thereto.

또 본 발명은 (E)-1-(2-하이드록시나프탈렌-1-일)-3-(2-메톡시페닐)프로프-2-엔-1-온을 용매에 녹여서 소듐하이드로옥사이드와 과산화수소를 첨가하고 그 반응용액을 여과하는 단계를 포함하는 하기 화학식 1의 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온 제조방법을 제공한다.The present invention also relates to a process for the preparation of (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2-methoxyphenyl) prop- Hydroxy-3- (2-methoxyphenyl) -1H-benzo [f] chromen-1-one represented by the following general formula (1), which comprises the steps of:

Figure 112012085906612-pat00005
Figure 112012085906612-pat00005

[화학식 1][Chemical Formula 1]

본 발명의 조성물은 다양한 담체 및 전달 시스템으로 제형화할 수 있다. 투여되는 치료 화합물의 양 및 화합물의 농도는 선택되는 비히클 또는 기기, 환자의 임상 상태, 부작용 및 화합물의 제형 안정성에 의존적이다. 따라서, 의사는 치료 화합물의 적정 농도를 포함하는 적정 조제물을 이용하며, 문제가 되는 환자나 유사한 환자에 대한 임상 경험에 따라 투여 제형의 양을 선정한다. The compositions of the present invention may be formulated into a variety of carriers and delivery systems. The amount of therapeutic compound administered and the concentration of the compound will depend on the vehicle or device selected, the clinical condition of the patient, side effects and the formulation stability of the compound. Thus, the physician will use the appropriate formulation containing the appropriate concentration of the therapeutic compound, and will select the dosage form according to clinical experience with the patient or similar patient in question.

또한, 제형에 부형제를 사용할 수 있다. 그 예로, 공-용매, 계면활성제, 오일, 습윤제, 에몰리언트, 보존제,안정화제 및 항산화제를 포함한다. 약학적으로 허용가능한 완충제로, 예컨대 트리 또는 포스페이트 완충제를 사용할 수 있다. 유효량의 희석제, 첨가제 및 부형제는 가용성, 생물 활성 등의 측면에서 약학적으로 허용가능한 제형을 수득하는데 유효한 양이다.In addition, excipients may be used in the formulation. Examples include co-solvents, surfactants, oils, wetting agents, emollients, preservatives, stabilizers and antioxidants. As a pharmaceutically acceptable buffer, for example, a tri or phosphate buffer may be used. Effective amounts of diluents, additives and excipients are amounts effective to obtain pharmaceutically acceptable formulations in terms of solubility, bioactivity, and the like.

그러므로, 본 발명의 조성물은 국소, 경구 또는 비경구 투여용 기존의, 약학적으로 허용가능한 비히클과 제형화될 수 있는 치료 화합물을 포함한다. 또한, 제형은 등장성, 생리학적 및 pH 안정성을 유지하기 위한, 완충제 및 보존제와 같은 소량의 보강제를 포함할 수 있다.Thus, the compositions of the present invention include conventional, pharmaceutically acceptable vehicles for topical, oral or parenteral administration, and therapeutic compounds that can be formulated. The formulations may also contain minor amounts of adjuvants, such as buffers and preservatives, to maintain isotonicity, physiological and pH stability.

본 발명의 조성물은 인간 및 동물 대상자 모두에 투여될 수 있다.The composition of the present invention can be administered to both human and animal subjects.

본 발명의 조성물은 활성 화합물이 하나 이상의 불활성 성분과 선택적으로 하나 이상의 부가적인 활성 성분과 밀접하게 혼합된, 조성물로 투여될 수 있다. 조성물은 당업계에 공지된 인간 및 동물 투여용 임의 조성물로 사용될 수 있다.The compositions of the present invention may be administered in a composition wherein the active compound is intimately mixed with one or more inactive ingredients and optionally one or more additional active ingredients. The compositions can be used as any composition for human and animal administration known in the art.

본 발명의 조성물은 투약 형태에 따라 적절한 경로에 따라 투여할 수 있다. 예로, 정맥내, 동맥내, 피하, 근육내 등으로 주사물을 투여할 수 있다.The composition of the present invention may be administered according to a proper route depending on the dosage form. For example, the subject can be administered intravenously, intraarterially, subcutaneously, intramuscularly, and the like.

경구 투여를 위해, 고체 또는 유체 단위 투약 형태 중 어느 하나를 제조할 수 있다. 수용성 형태는 당, 방향성 향미제 및 보존제와 함께 수성 비히클에 용해시켜 시럽을 제조할 수 있다. 방향성 향미제, 당 및 사카린과 같은 적정 감미제 및 하이드로-알콜성(예, 에탄올) 비히클을 이용하여, 엘리시르를 제조한다. 수성 비히클을, 아카시아, 트라가칸트, 메틸셀룰로스 등과 같은 현탁제를 이용하여, 현탁물을 제조할 수 있다. 본 발명의 합성 화합물은, 안정화제, 예컨대 에틸렌디아민테트라아세트산(EDTA)과 같은 금속 킬레이터 환원제, 또는 소듐 메타바이설파이트와 같은 환원제를 이용하여 제형화할 수 있다.For oral administration, either solid or fluid unit dosage forms may be prepared. The aqueous form may be dissolved in an aqueous vehicle together with a sugar, an aromatic flavor and a preservative to produce a syrup. Elsys are prepared using suitable sweeteners such as aromatic flavors, sugars and saccharin, and hydro-alcoholic (e.g., ethanol) vehicles. Suspensions can be prepared using aqueous vehicles, such as acacia, tragacanth, methylcellulose, and the like. The synthetic compound of the present invention may be formulated with a stabilizing agent, for example, a metal chelator reducing agent such as ethylenediaminetetraacetic acid (EDTA), or a reducing agent such as sodium metabisulfite.

비경구용 적정 제형은 당업계 실무자에게 자명하다. 일반적으로, 치료 화합물은 약 1 내지 약 100 mg/mL의 농도로 수용액 중에 제조된다. 보다 전형적으로, 농도는 약 10 내지 60 mg/mL 또는 약 20 mg/mL이다. 이용하기위해 선택된 화합물의 안정성 및 효능에 의존적으로, 일부 경우에서는 1 mg/mL 이하의 농도가 필수적일 수 있다.Suitable formulations for parenteral administration will be apparent to those skilled in the art. Generally, the therapeutic compound is prepared in aqueous solution at a concentration of about 1 to about 100 mg / mL. More typically, the concentration is about 10 to 60 mg / mL or about 20 mg / mL. Depending on the stability and efficacy of the compound selected for use, in some cases concentrations below 1 mg / mL may be necessary.

본 발명의 활성 성분의 유효 용량은 적어도 치료 조건의 성질, 독성, 그 화합물이 예방적으로(더 적은 용량으로) 사용되는지 또는 활성 암 감염에 대하여 사용되는지 여부, 전달 방법 및 그 약학 제형에 달려있으며, 통상의 용량 증가 연구를 사용하는 임상의에 의하여 결정될 것이다. 상기 용량은 1일에 약 0.0001 내지 약100 mg/몸무게kg, 일반적으로 1일에 약 0.01 내지 약 10mg/몸무게kg, 더 일반적으로 1일에 약 0.01 내지 약 5 mg/몸무게kg, 더 일반적으로 1일에 약 0.05 내지 약5 mg/몸무게 kg일 것으로 예상할 수 있다. 예컨대, 몸무게가 약70 kg인 성인을 위한 1일 희망 용량은 1 mg 내지 1000 mg, 바람직하기로는 5mg 내지 500 mg일 것이며, 단일 또는 다중 투여형일 수 있다.The effective dose of the active ingredient of the present invention will depend, at least, on the nature of the therapeutic condition, toxicity, whether the compound is used prophylactically (with less dosage) or against an active cancer infection, on the delivery method and on its pharmaceutical formulations , Will be determined by the clinician using conventional dose escalation studies. The dose is generally from about 0.0001 to about 100 mg / kg body weight per day, generally from about 0.01 to about 10 mg / kg body weight per day, more usually from about 0.01 to about 5 mg / kg body weight per day, more usually from 1 Day to about 0.05 mg / kg body weight per day. For example, the daily recommended dose for an adult weighing about 70 kg will be from 1 mg to 1000 mg, preferably from 5 mg to 500 mg, and may be single or multiple dosage forms.

멸균 제형이, 진피내, 관절내, 근육내, 혈관내, 정맥내, 흡입 및 피하를 포함한 다양한 비경구 경로에 적합하다.The sterile formulations are suitable for a variety of parenteral routes including intraperitoneal, intraarticular, intramuscular, intravascular, intravenous, inhalation and subcutaneous.

다수의, 바이오폴리머(생물 기반 시스템), 리포좀 시스템 및 폴리머 전달 시스템, 예컨대 덴드리머 중 임의의 것을 포함하는 느린 또는 연장된 방출 전달 시스템은, 본원의 조성물에 활용하여, 치료 화합물의 연속적인 또는 장기적인 소스를 제공할 수 있다. 이러한 느린 방출 시스템은 국소, 눈, 경구 및 비경구용 제형으로 적용가능하다.A slow or extended release delivery system comprising any of a number of biopolymers (bio-based systems), liposome systems and polymer delivery systems, such as dendrimers, can be utilized in the compositions herein to provide a continuous or long term source Can be provided. Such slow release systems are applicable for topical, ocular, oral and parenteral formulations.

본 발명의 합성 화합물(들)은 또한 뉴트라-약제(nutrapharmaceutical) 또는 식품의약(nutraceutical)으로서 제형화될 수 있다. 예컨대, 합성 화합물(들)은 시리얼, 과일 쥬스와 같은 음료, 알콜 음료, 빵 등과 같은 경구 식이용 식품으로 제형화할 수 있다.The synthetic compound (s) of the present invention may also be formulated as a nutraceutical or a nutraceutical. For example, the synthetic compound (s) can be formulated into oral use foods such as cereal, fruit juices, alcoholic beverages, bread, and the like.

이하 본 발명을 설명한다.Hereinafter, the present invention will be described.

본 발명은 효과적인 항암제를 찾기 위하여 벤조플라보놀 유도체를 고안하여 합성하고 이들에 대한 대장암 세포의 세포사멸 유도 효과를 관찰하였다. 벤조플라보놀 유도체는 대장암세포의 세포사멸을 유도한다.In order to find an effective anticancer agent, the present inventors designed and synthesized benzoflavone derivatives, and observed the effect of inducing apoptosis on colon cancer cells. The benzoflavone derivatives induce apoptosis of colon cancer cells.

본 발명에서는 베타벤조플라본에 히드록시기와 메톡시기를 각각 치환시킨 벤조플라본 유도체를 고안하여 합성하였다. 히드록시기가 3번 위치에 치환되어 결합할 때 특별히 플라보놀이라고 부르는데 본 발명의 플라본노이드 유도체 역시 3번 위치에 히드록시기를 치환한 구조이므로 베타벤조플라보놀로 불리는 물질이 고안되었다. 이 물질이 보이는 항암효과를 측정한 결과 우수한 효과를 보였기 때문에 베타벤조플라보놀 유도체가 항암제로 활용이 가능하다고 판단되어 본 발명을 완성하였다.In the present invention, a benzoflavone derivative substituted with a hydroxy group and a methoxy group was prepared and synthesized. When the hydroxy group is substituted at the 3-position, it is particularly called as flavanol. Since the flavonoid derivative of the present invention also has a structure in which the hydroxyl group is substituted at the 3-position, a substance called beta-benzoflavonol was devised. As a result of measuring the anticancer effect of this substance, it was determined that beta-benzoflavone derivative could be used as an anticancer drug because of its excellent effect.

본 발명의 벤조플라보놀 유도체는 대장암 세포의 세포사멸 유도 효과를 보임으로써 암 질환의 예방 및 치료를 위한 약학조성물로 유용하게 이용될 수 있다.The benzoflavone derivative of the present invention exhibits cell death-inducing effect of colon cancer cells, and thus can be usefully used as a pharmaceutical composition for prevention and treatment of cancer diseases.

도 1은 벤조플라보놀 유도체 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온 (DK98)의 수소핵자기공명분광스펙트럼이다. (400MHz 브루커 핵자기공명분광기기 사용)
도 2는 벤조플라보놀 유도체 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온 (DK98)의 탄소핵자기공명분광스펙트럼이다. (100MHz 브루커 핵자기공명분광기기 사용)
도 3은 벤조플라보놀 유도체 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온 (DK98)의 고분해능질량분석 스펙트럼이다. (제올사 (Jeol Ltd., Tokyo, Japan)의 JMS700 HREIMS (high-resolution electron impact ionization mass spectrometer) 기기 사용)
도 4는 벤조플라보놀 유도체 (DK98)의 대장암 세포 성장 억제 효과이다.
도 5는 대장암 세포에서 벤조플라보놀 유도체 (DK98)의 카스파제-7 활성 효과이다.
도 6은 벤조플라보놀 유도체 (DK98)에 의한 대장암 세포의 세포사멸 유도 효과이다. 1 is a hydrogen nuclear magnetic resonance spectroscopy of the benzoflavone derivative 2-hydroxy-3- (2-methoxyphenyl) -1H-benzo [f] chromen-1-one ( DK98 ). (Using a 400 MHz Bruker nuclear magnetic resonance spectrometer)
2 is a carbon nuclear magnetic resonance spectroscopy spectrum of the benzoflavone derivative 2-hydroxy-3- (2-methoxyphenyl) -1H-benzo [f] chromen-1-one ( DK98 ). (Using a 100 MHz Bruker nuclear magnetic resonance spectrometer)
3 is a high-resolution mass spectrometry spectrum of the benzoflavone derivative 2-hydroxy-3- (2-methoxyphenyl) -1H-benzo [f] chromen-1-one ( DK98 ). (Using a JMS700 HREIMS (high-resolution electron impact ionization mass spectrometer) instrument from Jeol Ltd., Tokyo, Japan)
4 is an inhibitory effect of benzoflavone derivative ( DK98 ) on growth of colon cancer cells.
5 is an effect of caspase-7 activity of the benzoflavone derivative ( DK98 ) in colorectal cancer cells.
6 is a cell-killing induction effect of a benzoflavone derivative ( DK98 ).

이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

실시예Example 1.  One. 벤조플라보놀Benzoflavone 유도체의 합성 Synthesis of derivatives

본 발명의 벤조플라보놀 유도체는 아래 반응식에 나타낸 방법을 사용하여 다음과 같이 합성하였다.The benzoflavone derivatives of the present invention were synthesized as follows using the method shown in the following reaction formula.

Figure 112012085906612-pat00006
Figure 112012085906612-pat00006

화합물 I은 (E)-1-(2-hydroxynaphthalen-1-yl)-3-(2-methoxyphenyl)Compound I was prepared from (E) -1- (2-hydroxynaphthalen-1-yl) -3- (2-methoxyphenyl)

prop-2-en-1-one로서 J. Indian Chem. Soc. 49: 725 (1972)논문에 이미 발표된 방법을 따라서 준비하였다, 화합물 1을 9 ㎖의 메탄올 용매에서 교반하며 소듐하이드로옥사이드 (NaOH, 260mg, 6.5mmol)과 30% 과산화수소 (0.5ml)를 증류수 7 ml와 함께 넣는다. 10시간 동안 상온에서 교반 시킨다. 얇은막 크로마토그라피를 통해 반응 종료를 확인하고, 반응용액에 얼음물 50 ml를 반응물에 넣는다. 클로로폼 용매를 사용하여 30 ml씩 3번 추출하고 유기 용매를 감압 여과한다. 유기용매를 마그네슘 설페이트로 건조시키고 다시 감압여과한 후, 용매를 감압하에서 제거한 후, 생성된 잔사를 컬럼 크로마토그래피로 정제하여 목적화합물 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온 (2-hydroxy-3- (2-methoxyphenyl)-1H-benzo[f]chromen-1-one ; DK98)를 80 mg (25 %)의 수율로 얻었다.prop-2-en-1-one as described in J. Indian Chem. Soc. 49: 725 (1972) paper were prepared according to previously published method for stirring the compounds (1) in a methanol solvent in 9 ㎖ and sodium hydroxide (NaOH, 260mg, 6.5mmol) and 30% hydrogen peroxide with distilled water for seven (0.5ml) ml. Stir at room temperature for 10 hours. The reaction is terminated by thin membrane chromatography and 50 ml of ice water is added to the reaction solution. Extract 3 times with 30 ml each time using chloroform solvent and filter the organic solvent under reduced pressure. The organic solvent was dried over magnesium sulfate and filtered again under reduced pressure. The solvent was removed under reduced pressure, and the resulting residue was purified by column chromatography to obtain the target compound 2-hydroxy-3- (2-methoxyphenyl) Benzo [f] chromen-1-one; DK98 ) was obtained in a yield of 80 mg (25%).

최종산물의 확인을 위해 핵자기공명분광 실험을 수행하였다. 사용한 기기는 브루커사 400MHz 기기였다. 또한 핵자기공명분광법으로 확인한 유도체의 구조를 재확인하기 위하여 고분해능질량분석법을 이용하였다. 사용한 기기는 제올사 (Jeol Ltd., Tokyo, Japan)의 JMS700 HREIMS (high-resolution electron impact ionization mass spectrometer) 였다. 핵자기공명분광법으로 확인 유도체의 수소와 탄소의 위치에 따른 명명법은 아래 [화학식 2]의 번호를 따랐다.Nuclear magnetic resonance spectroscopy experiments were performed to confirm the final product. The device used was a Bruscia 400MHz device. High resolution mass spectrometry was also used to confirm the structure of the derivatives identified by nuclear magnetic resonance spectroscopy. The instrument used was JMS700 HREIMS (high-resolution electron impact ionization mass spectrometer) from Jeol Ltd., Tokyo, Japan. Nuclear magnetic resonance spectroscopy was used to follow the nomenclature of the confirmed derivatives according to the positions of hydrogen and carbon.

[화학식 2](2)

Figure 112012085906612-pat00007
Figure 112012085906612-pat00007

벤조플라보놀 유도체 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온 (2-hydroxy-3-(2-methoxyphenyl)-1H-benzo[f]chromen-1-one ; DK98)의 수소핵자기공명분광스펙트럼과 탄소핵자기공명스펙트럼은 각각 도 1과 도 2에 나타낸 바와 같고 화학적이동도는 아래와 같다.Benzoflavone derivative 2-hydroxy-3- (2-methoxyphenyl) -1H-benzo [f] chromen-1-one; DK98 ) are shown in FIG. 1 and FIG. 2, respectively, and the chemical mobility thereof is as follows.

1H NMR (400MHz, DMSO-d6) δ 10.02 (d, 1H, H-5a, J = 8.5 Hz), 9.01 (s, 1H, -OH), 8.27 (d, 1H, H-7, J = 9.1 Hz), 8.10 (d, 1H, H-6a, J = 7.9 Hz), 7.82 (m, 1H, H-5b), 7.69 (m, 1H, H-6b), 7.68 (d, 1H, H-8, J = 9.1 Hz), 7.56 (d, 1H, H-6', J = 7.5 Hz), 7.54 (m, 1H, H-4'), 7.23 (d, 1H, H-3' J = 8.3 Hz), 7.11 (t, 1H, H-5', J = 7.5 Hz), 3.82 (s, 3H, 2'-OCH3); 13C NMR (100MHz, DMSO-d6) δ 174.1 (C-4), 157.2 (C-2'), 156.2 (C-9), 144.1 (C-2), 141.0 (C-3), 135.1 (C-7), 131.8 (C-4'), 131.2 (C-6'), 129.9 (C-5), 129.8 (C-6), 129.0 (C-5b), 128.6 (C-6a), 126.4 (C-6b), 125.8 (C-5a), 120.2 (C-5'), 119.5 (C-1'), 118.2 (C-8), 114.3 (C-10), 112.0 (C-3'), 55.8 (C-2'-OCH3). 1 H NMR (400MHz, DMSO- d 6) δ 10.02 (d, 1H, H-5a, J = 8.5 Hz), 9.01 (s, 1H, -OH), 8.27 (d, 1H, H-7, J = 9.1 Hz), 8.10 (d, 1H, H-6a, J = 7.9 Hz), 7.82 (m, 1H, H-5b), 7.69 (m, 1H, H-6b), 7.68 (d, 1H, H- 8, J = 9.1 Hz), 7.56 (d, 1H, H-6 ', J = 7.5 Hz), 7.54 (m, 1H, H-4'), 7.23 (d, 1H, H-3 'J = 8.3 Hz), 7.11 (t, 1H , H-5 ', J = 7.5 Hz), 3.82 (s, 3H, 2'-OCH 3); 13 C NMR (100MHz, DMSO- d 6) δ 174.1 (C-4), 157.2 (C-2 '), 156.2 (C-9), 144.1 (C-2), 141.0 (C-3), 135.1 ( C-7), 131.8 (C-4 '), 131.2 (C-6'), 129.9 (C-6), 125.8 (C-5a), 120.2 (C-5 '), 119.5 (C-1'), 118.2 , 55.8 (C-2'-OCH 3).

벤조플라보놀 유도체 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온 (DK98)은 현재까지 보고되지 않은 새로운 물질로서 C20H14O4의 분자식을 갖는다. 핵자기공명분광법으로 확인한 이 화합물의 구조를 확증하기 위하여 고분해능질량분석법을 이용하였고, 이론적인 분자량이 318.0892이었고 실험으로 얻은 분자량은 318.0989이었기 때문에 이 화합물은 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온으로 확인되었다. 이 화합물의 고분해능질량분석 스펙트럼은 도 3과 같다.Of benzo flavonol derivatives 2-hydroxy-3- (2-methoxy-phenyl) -1H- benzo [f] chromene-1-one (DK98) is a new material that is not reported to date C 20 H 14 O 4 Have molecular formula. High-resolution mass spectrometry was used to confirm the structure of this compound as confirmed by nuclear magnetic resonance spectroscopy. Since the theoretical molecular weight was 318.0892 and the molecular weight obtained by the experiment was 318.0989, the compound was 2-hydroxy-3- (2- Phenyl) -lH-benzo [f] chromen-l-one. The high-resolution mass spectrometry spectrum of this compound is shown in FIG.

실시예Example 2.  2. 벤조플라보놀Benzoflavone 유도체 ( Derivatives ( DK98DK98 )의 대장암 세포의 세포성장 억제 효과) Inhibited the growth of colon cancer cells

HCT116 대장암 세포를 ATTC(American Type Culture Collection)로부터 구입하여 10% FBS (Fetal Bovine Serum, Invitrogen Life Technologies), Antibiotic-Antimycotic solution (Invitrogen Life Technologies) 포함한 DMEM (Invitrogen Life Technologies) 배양액을 2일에 한 번씩 100-mm 세포배양접시에 1 x 106의 접종 밀도(seed density)로 계대하면서 37℃, 5% CO2 배양기에서 배양하였다. 벤조플라보놀 유도체 (DK98)에 의한 세포성장 억제 효과는 암세포의 콜로니 형성능 평가 (Colonony forming assay)를 통하여 세포 성장이 억제되는지의 여부로 측정하였다. HCT116 대장암 세포를 24-well 배양접시에 well 당 6000개 세포로 분주한 후 0, 5, 10, 20 μM 농도의 벤조플라보놀 유도체 (DK98)을 처리하고, 7일 후 6% 글루타르알데하이드 (glutaraldehyde)와 0.5% 크리스탈바이올렛 (crystal violet) 용액을 1:1로 섞어준 혼합액을 세포에 첨가한 후 15분 동안 반응시켜 남아있는 세포를 염색하였다. 그 결과 도 4에 나타난 바와 같이 벤조플라보놀 유도체 (DK98)을 10 μM 농도 이상으로 처리했을 때 암세포의 콜로니 형성능이 급격히 감소되는 것이 관찰되었다. 이러한 사실로부터 벤조플라보놀 유도체 (DK98)은 대장암 세포의 성장을 억제시키는 효과가 있다는 사실을 확인하였다.
HCT116 colon cancer cells were purchased from the American Type Culture Collection (ATTC) and DMEM (Invitrogen Life Technologies) culture medium containing 10% FBS (Fetal Bovine Serum, Invitrogen Life Technologies) and Antibiotic-Antimycotic solution The cells were cultured in a 5% CO 2 incubator at 37 ° C in a 100-mm cell culture dish at a seeding density of 1 × 10 6 . The inhibitory effect of the benzoflavone derivative (DK98) on cell growth was determined by whether the cell growth was inhibited by the colony forming assay of cancer cells. HCT116 colon cancer cells were treated with 6000 cells per well in a 24-well culture dish, treated with benzoflavone derivative (DK98) at concentrations of 0, 5, 10 and 20 μM, and treated with 6% glutaraldehyde glutaraldehyde) and 0.5% crystal violet (1: 1) were added to the cells, followed by reaction for 15 minutes to stain the remaining cells. As a result, as shown in FIG. 4, when the benzoflavone derivative (DK98) was treated at a concentration of 10 μM or more, it was observed that the colony forming ability of cancer cells was drastically reduced. From these facts, it has been confirmed that the benzoflavone derivative (DK98) has an effect of inhibiting the growth of colon cancer cells.

실시예Example 3.  3. 벤조플라보놀Benzoflavone 유도체 ( Derivatives ( DK98DK98 )의 세포사멸 유도 ) Induced apoptosis 카스파제Caspase -7 효소 활성 효과-7 enzyme activity effect

본 발명의 벤조플라보놀 유도체 (DK98) 화합물에 의해 세포사멸 유도 단백질인 카스파제-7 효소의 활성이 증가되는지 여부를 분석하기 위하여, DK98 화합물을 처리한 HCT116 대장암 세포주에서 활성형태의 절단형 카스파제-7 단백질 및 이 효소의 기질 단백질인 poly(ADP-ribose)polymerase (PARP) 단백질의 절단 변화를 면역블롯법을 이용하여 조사하였다. HCT116 대장암 세포를 60-㎜ 배양접시에서 1.5 X 106개 되도록 배양하고, 20 μM 농도가 되도록 DK98 화합물을 처리한 후 0, 24, 48시간 경과 후 세포를 수확하여, 세포용해액(cell lysis buffer)으로 세포를 용해시킨 후, 고속원심분리하여 세포 용해액만을 수확하였다. 동량의 단백질이 포함하도록 제조된 시료를 SDS-폴리아크릴아마이드 겔(SDS-polyacrylamide gel) 전기영동을 실시하여 세포에 존재하는 단백질들을 분리하였다. 전기영동으로 분리된 단백질을 폴리스틸렌 막 (polystyrene membrane)으로 옮긴 후 절단형의 카스파제-7 단백질 및 PARP와 결합하는 일차 항체(Cell Signaling Technology 회사에서 구입)와 대조군으로서 단백질 발현이 변화되지 않는 GAPDH를 인지하는 일차항체 (Santa Cruz technology 회사에서 구입)를 각각 5시간 반응시킨 후, 일차항체를 인지하는 이차항체 (Cell Signaling Technology 회사에서 구입)를 1시간동안 반응시켰다. 화학형광감지 시스템 (chemiluminescence) (Amersham Pharmacia Biotechnology에서 구입)을 이용하여 각종 단백질 활성형을 분석하였다. In order to analyze whether the activity of the caspase-7 enzyme, which is a cell death-inducing protein, was increased by the benzoflavone derivative (DK98) compound of the present invention, in the HCT116 colon cancer cell line treated with the DK98 compound, The cleavage of poly (ADP-ribose) polymerase (PARP) protein, a substrate protein of this enzyme, was investigated by immunoblotting. HCT116 colon cancer cells were cultured in a 60-mm culture dish to 1.5 × 10 6 cells, treated with DK98 compound to a concentration of 20 μM, and after 0, 24 and 48 hours, the cells were harvested and cell lysis buffer), followed by high-speed centrifugation to harvest only the cell lysate. Samples prepared to contain the same amount of protein were subjected to SDS-polyacrylamide gel electrophoresis to separate the proteins present in the cells. The primary antibody (purchased from Cell Signaling Technology), which binds cleaved form of caspase-7 protein and PARP after transferring the separated proteins to a polystyrene membrane, and GAPDH that does not change protein expression as a control (Purchased from Santa Cruz Technology) were reacted for 5 hours each, and then a secondary antibody (purchased from Cell Signaling Technology), which recognizes the primary antibody, was reacted for 1 hour. Various protein activity types were analyzed using a chemiluminescence detection system (purchased from Amersham Pharmacia Biotechnology).

도 5에 나타낸 바와 같이 DK98 화합물을 HCT116 암세포에 처리하면 대조 단백질인 GAPDH의 양은 변하지 않았으나, 처리 24 시간 후부터 세포사멸을 유도하는 단백분해 효소인 카스파제-7의 잘려진 활성형이 축적되기 시작하였으며, 카스파제-7의 기질 단백질인 PARP 단백질의 절단이 증가되었다. 이러한 결과는 본 발명의 DK98 화합물이 대장암 세포에서 카스파제 효소 활성을 통하여 세포사멸을 유도함으로써 항암 효과를 나타낸다는 사실을 시사하는 것이다.
As shown in FIG. 5, when the DK98 compound was treated with HCT116 cancer cells, the amount of GAPDH as a control protein was not changed. However, after 24 hours of treatment, the cleaved active form of caspase-7, a proteolytic enzyme that induces apoptosis, The cleavage of PARP protein, a substrate protein of caspase-7, was increased. These results suggest that the DK98 compound of the present invention induces apoptosis through colonization of caspase enzyme activity in colorectal cancer cells.

실시예Example 4.  4. 벤조플라보놀Benzoflavone 유도체 ( Derivatives ( DK98DK98 )의 세포사멸 유도 효과) Induced apoptosis

카스파제-7 효소 활성을 유도하는 본 발명의 벤조플라보놀 유도체 (DK98)가 대장암세포의 세포사멸을 유도하는지 분석하였다. 일반적으로 세포사멸이 유도되는 세포에서는 세포막 내부에 존재하는 인지질 phosphatidylserine (PS)이 바깥쪽 세포막으로 이동한다. Annexin V 단백질은 PS와 강한 친화력을 보이는 단백질로서 세포와 반응시키면 세포사멸이 일어나고 있는 세포에만 결합한다. 이러한 원리를 이용하여 형광을 붙인 Annexin V 단백질 (Anexin V-FITC)과 약물을 처리한 세포를 반응시키고, 형광을 측정하면 세포사멸이 일어난 세포를 정량적으로 분석할 수 있게 된다. 본 발명의 벤조플라보놀 유도체 (DK98) 화합물의 세포사멸 유도 효과를 측정하기 위하여 HCT116 대장암세포에 20 μM 농도의 벤조플라보놀 유도체 (DK98) 화합물 처리하고 2일 후 1% 트립신-EDTA 용액을 첨가해 배양 용기에 부착되어 있던 세포를 떼어내어 세포를 수확하였다. 수확된 세포에 10 μg/ml 농도의 PI (Propidium Iodide)와 Annexin V-FITC (덴마크 Chemometec 회사에서 구입) 용액 2 μl를 첨가한 후 15분간 반응시키고 유세포 분리측정기 (NucleoCounter NC-3000, Chemometec 회사, 덴마크)를 이용하여 형광이 나타나는 세포를 측정하였다.It was analyzed whether the benzoflavone derivative (DK98) of the present invention which induces caspase-7 enzyme activity induces apoptosis of colon cancer cells. In general, phosphatidylserine (PS), which is present inside the cell membrane, migrates to the outer cell membrane in cells where cell death is induced. Annexin V protein, a protein with strong affinity to PS, binds only to cells that undergo apoptosis when reacted with cells. Using this principle, the fluorescently labeled Annexin V protein (Anexin V-FITC) is reacted with the drug-treated cells and fluorescence can be quantitatively analyzed. To determine the cell-killing effect of the benzoflavone derivative (DK98) compound of the present invention, HCT116 colon cancer cells were treated with a 20 μM benzoflavone derivative (DK98) compound and 2 days later, 1% trypsin-EDTA solution was added The cells attached to the culture container were removed and the cells were harvested. To the harvested cells, 2 μl of a solution of PI (Propidium Iodide) and Annexin V-FITC (purchased from Chemometec Company, Denmark) at a concentration of 10 μg / ml was added and reacted for 15 minutes. The cells were reacted with a flow cytometer (NucleoCounter NC- Denmark) was used to measure the fluorescence of the cells.

도 6에 나타낸 바와 같이, 정상적으로 성장하고 있는 HCT116 대장암세포의 21% 정도가 Annexin V와 반응하였지만, 본 발명의 벤조플라보놀 유도체 (DK98) 화합물을 처리한 세포에서는 86% 세포에서 Annexin V와 결합하였다. 이러한 사실은 벤조플라보놀 유도체 (DK98) 화합물을 HCT116 대장암세포에 처리하면 세포사멸이 촉진된다는 사실을 의미하는 것이다.As shown in Fig. 6, about 21% of the normally growing HCT116 colon cancer cells reacted with Annexin V, but in the cells treated with the benzoflavone derivative (DK98) compound of the present invention, they were bound to Annexin V in 86% . This fact implies that treatment of the benzoflavone derivative (DK98) compound with HCT116 colon cancer cells promotes apoptosis.

이상, 본 발명의 내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 구현 예일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.
Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

삭제delete 삭제delete 삭제delete 삭제delete 삭제delete 하기 화학식 1의 화합물 및 약학적으로 허용가능한 담체를 유효성분으로 포함하는 대장암 예방 및 치료용 약학 조성물.
Figure 112014007192200-pat00011

[화학식 1]A pharmaceutical composition for the prevention and treatment of colorectal cancer, comprising a compound of the following formula (I) and a pharmaceutically acceptable carrier as an active ingredient.
Figure 112014007192200-pat00011

[Chemical Formula 1] 삭제delete (E)-1-(2-하이드록시나프탈렌-1 -일)-3-(2-메톡시페닐)프로프-2-엔-1-온을 용매에 녹여서 소듐하이드로옥사이드와 과산화수소를 첨가하고 그 반응용액을 여과하는 단계를 포함하는 제조방법에 의하여 제조된 하기 화학식 1의 2-히드록시-3-(2-메톡시페닐)-1H-벤조[f]크로멘-1-온을 유효성분으로 하는 대장암 예방 및 치료용 약학 조성물.
Figure 112014007192200-pat00012

[화학식 1]
(E) -1- (2-hydroxynaphthalen-1-yl) -3- (2-methoxyphenyl) prop-2-en-1-one was dissolved in a solvent to which sodium hydroxide and hydrogen peroxide were added 2-hydroxy-3- (2-methoxyphenyl) -1H-benzo [f] chromen-1-one represented by the following formula 1 prepared by a process comprising the steps of: Which comprises administering to a mammal in need thereof an effective amount of a compound of formula (I).
Figure 112014007192200-pat00012

[Chemical Formula 1]
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007115181A2 (en) 2006-04-03 2007-10-11 Eastman Chemical Company Compounds exhibiting efflux inhibitor activity and compositions and uses thereof
KR20090027412A (en) * 2007-09-12 2009-03-17 건국대학교 산학협력단 Novel cytotoxic flavone derivative 7-o-(3-benzyloxypropyl)5,4 -di-o-methylapigenin, the preparation method and composition for treating cancers comprising the compound
KR20110139397A (en) * 2010-06-23 2011-12-29 건국대학교 산학협력단 5,6-benzoflavone having multidrug resistance inhibitory activity, a method for preparing the same and a pharmaceutical composition comprising the same

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007115181A2 (en) 2006-04-03 2007-10-11 Eastman Chemical Company Compounds exhibiting efflux inhibitor activity and compositions and uses thereof
KR20090027412A (en) * 2007-09-12 2009-03-17 건국대학교 산학협력단 Novel cytotoxic flavone derivative 7-o-(3-benzyloxypropyl)5,4 -di-o-methylapigenin, the preparation method and composition for treating cancers comprising the compound
KR20110139397A (en) * 2010-06-23 2011-12-29 건국대학교 산학협력단 5,6-benzoflavone having multidrug resistance inhibitory activity, a method for preparing the same and a pharmaceutical composition comprising the same

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Title
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Bioorganic & Medicinal Chemistry. 2006. Vol. 14, pp. 2966-2971*

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