KR20040110119A - Composition for curing cancer containing the extract of Jipaesan - Google Patents
Composition for curing cancer containing the extract of Jipaesan Download PDFInfo
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- KR20040110119A KR20040110119A KR1020030039292A KR20030039292A KR20040110119A KR 20040110119 A KR20040110119 A KR 20040110119A KR 1020030039292 A KR1020030039292 A KR 1020030039292A KR 20030039292 A KR20030039292 A KR 20030039292A KR 20040110119 A KR20040110119 A KR 20040110119A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/232—Angelica
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8966—Fritillaria, e.g. checker lily or mission bells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
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- General Health & Medical Sciences (AREA)
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Abstract
Description
본 발명은 지패산 추출물을 함유하는 암 치료용 조성물에 관한 것으로, 보다 구체적으로는 백지(白芷,Angelica Dahurica)와 패모(貝母,Fritillariae Verticillata)로 조성된 지패산(芷貝散, Jipaesan)의 가용성 추출물을 함유함으로써 암세포, 특히 인간 백혈병 세포주(Human Leukemia Cells)에 대한 세포증식 억제, 세포분화 촉진 또는 아폽토시스 촉진 등의 뛰어난 효능이 있는 암 치료용 조성물에 관한 것이다.The present invention relates to a composition for treating cancer containing Zipacic Acid extract, more specifically, of Jipaesan composed of white paper (白芷, Angelica Dahurica ) and Femo (貝母, Fritillariae Verticillata ) By containing a soluble extract, the present invention relates to a composition for treating cancer with excellent efficacy such as inhibiting cell proliferation, promoting cell differentiation, or promoting apoptosis on cancer cells, particularly human leukemia cells.
현대 과학기술의 발달과 더불어 의학기술, 생명공학 및 유전공학이 발전함에 따라 암을 치료하기 위한 다양한 연구 및 방법들이 개발되고 있음에도 불구하고, 암은 아직 완치될 수 없는 중대한 질환으로 인식되고 있다. 이러한 암을 치료하기 위한 방법으로 절개술, 화학요법, 방사선요법, 절개술 등의 많은 방법이 개발되었지만 방사선 요법이나 절개술 등은 주로 암의 초기진단이 이루어졌을 때에만 효과가 있을 뿐 말기암으로 진행된 상태에서는 소용이 없으며 화학요법을 주로 사용하여 치료할 수 밖에 없다. 그러나, 그 완치율은 상당히 낮은 실정이다.Despite the development of modern science and technology along with the development of medical technology, biotechnology and genetic engineering, cancer has been recognized as a serious disease that cannot be cured yet. Many methods such as incision, chemotherapy, radiotherapy, and incision have been developed to treat these cancers. However, radiation therapy and incisions are effective only when the initial diagnosis of cancer is done. It is of no use and can only be treated with chemotherapy. However, the cure rate is quite low.
이에 따라, 임상분야에서는 기존의 항암제보다 항암 효능이 더욱 뛰어난 새로운 항암제를 요구하고 있으나, 이러한 항암제를 개발하기 위해서는 항암제의 새로운 표적에 대한 개발이 요구된다.Accordingly, the clinical field requires a new anticancer agent that is more effective than conventional anticancer agents, but in order to develop such anticancer agents, development of a new target of anticancer agents is required.
최근, 이러한 항암제의 표적으로 아폽토시스(apoptosis)에 관여하는 여러 유전자 및 단백질의 차단에 대한 연구가 진행되고 있을 뿐만 아니라 특정 세포, 특히 미분화되어 영구적으로 증식하는 암세포를 특정 기능을 발휘할 수 있는 세포로 분화시키기 위한 연구가 활발히 진행되고 있다. 이러한 연구들은 지난 수년동안 화학요법에 이용되었던 항암제들이 암세포의 유전자를 손상시켜 아폽토시스, 즉 계획된 세포 사멸(programmed cell death)을 유발시킨다는 것에 대한 발견에서부터 시작하여 사멸되지 않은 세포를 분화시키기 위한 세포분화 신호전달 조절 분자들을 발견하기에 이르렀다. 이에, 암치료에 대한 화학요법이 활발히 진행되고 있다.Recently, studies on the blocking of various genes and proteins involved in apoptosis as targets of such anticancer agents have been conducted, as well as differentiation of specific cells, especially undifferentiated and permanently proliferating cancer cells into cells capable of performing specific functions. There is an active research to make it possible. These studies begin with the discovery that chemotherapy drugs used in chemotherapy over the past several years can damage genes in cancer cells, leading to apoptosis, or programmed cell death, and to signal cell differentiation for differentiating unkilled cells. The discovery led to the discovery of delivery control molecules. Thus, chemotherapy for cancer treatment is actively progressing.
아폽토시스는 모든 세포에서 나타나는 세포 사멸의 한 형태로, 특히 바이러스, 박테리아 등에 감염된 세포(virus-infected cells) 및 자연적인 돌연변이에 의해 종양(tumor)으로 변형된 세포들을 효과적으로 사멸시키는 계획된 세포사멸이다. 아폽토시스를 통해 세포 사멸이 이루어지면 뚜렷한 형태적, 생화학적 특징이 나타난다. 또한, 세포분화는 비정상적으로 계속되는 세포 분열을 차단하고, 세포가 가지는 고유의 기능을 발휘할 수 있도록 형태적, 기능적으로 그 특징이 뚜렷히 나타난다.Apoptosis is a form of cell death that occurs in all cells, particularly planned cell death that effectively kills virus-infected cells and cells that have been transformed into tumors by natural mutations, particularly those infected by viruses, bacteria, and the like. Apoptosis results in cell death with distinct morphological and biochemical characteristics. In addition, cell differentiation is characterized by morphologically and functionally to block abnormally continuous cell division and to exhibit the inherent function of the cell.
비록, 아폽토시스를 통해 종양 세포들을 죽일 수 있다는 기전에는 논쟁의 여지가 있을지라도, 아폽토시스를 유발할 수 있는 물질을 찾는 것은 새로운 항암제의 동정방법으로 인식되고 있으며, 여기에 암세포의 가장 큰 특징인 계속적인 분열을차단하고 세포의 분화를 촉진시키는 물질을 발견하는 것은 곧 암을 치료하는데 이용될 수 있는 물질의 발견이라 할 수 있는 것이다.Although the mechanisms by which tumor cells can be killed through apoptosis are controversial, finding a substance that can induce apoptosis is recognized as a method for identifying new anticancer agents, which is one of the most distinctive features of cancer cells. Finding substances that block and promote cell differentiation is the discovery of substances that can be used to treat cancer.
따라서 새로운 항암 치료제를 개발하기 위해서는 상기와 같은 아폽토시스를 유발함과 동시에 사멸되지 않은 잔여 세포의 분화를 유도하는 물질의 탐색이 요구되는 것이다.Therefore, in order to develop a new anti-cancer drug, it is necessary to search for a substance that induces apoptosis as described above and induces differentiation of residual cells which are not killed.
아폽토시스 세포 사멸은 일반적으로 수명이 다한 일반 세포들에서 여러 유전자들의 작용에 의해 일어나며, 미분화 조세포(stem cell)로부터 새로운 세포가 생성되는 것만큼 중요하다. 아폽토시스를 통해 세포 사멸이 이루어지는 세포는 염증반응 등에 의해 인접한 세포에 어떠한 영향도 미치지 않으며, 계획된 사멸을 수행하여 자신만이 파괴된다. 이러한 아폽토시스의 생리적인 과정은 심각한 세포 손상과 팽배(swelling) 및 파손(lysis)이 일어난 후, 염증 반응을 유발하여 주위 세포들마저도 파괴하는 세포 괴사(necrotic cell death)와는 구별된다. 그러므로 화학요법에 있어서 암세포를 효과적으로 파괴하기 위해서는 특정 약물을 처리함으로써 아폽토시스 과정을 유도하여야 하는 것이다.Apoptosis cell death is usually caused by the action of several genes in end-of-life normal cells, and as important as new cells are produced from undifferentiated stem cells. Cells that undergo cell death through apoptosis do not have any effect on adjacent cells, such as by inflammatory reactions, and only destroy themselves by performing planned death. The physiological process of apoptosis is distinguished from necrotic cell death, which causes an inflammatory response and also destroys surrounding cells after severe cell damage, swelling and lysis. Therefore, in order to effectively destroy cancer cells in chemotherapy, the apoptosis process should be induced by treating certain drugs.
따라서, 아폽프토시스를 유도하고 세포분화를 통해 세포증식을 억제 할 수 있는 물질의 개발은 염증 반응을 통한 일반 세포의 사멸을 유도하지도 않으면서도, 효과적으로 종양세포만을 사멸시킬 수 있을 뿐만 아니라 잔여 암세포의 증식을 억제시키는 특징을 가질 수 있다.Therefore, the development of a substance capable of inducing apoptosis and inhibiting cell proliferation through cell differentiation can effectively kill only tumor cells as well as killing remaining cancer cells without inducing the death of normal cells through inflammatory reactions. It may have a characteristic of inhibiting proliferation.
이에, 효과적으로 아폽토시스를 유발하는 한편 세포증식 억제 및 세포분화를 촉진시키는 물질에 대한 요구가 증대되고 있는 실정이다.Accordingly, there is an increasing demand for substances that effectively induce apoptosis while promoting cell proliferation inhibition and cell differentiation.
상기와 같은 요구가 증대됨에 따라 암세포의 아폽토시스를 유발하고 세포증식을 억제하고 세포분화를 촉진하는 특성이 있는 물질을 개발하기 위한 연구를 지속적으로 수행한 결과, 본 발명자들은 백지로부터 분리한 가용성 추출물은 세포의 아폽토시스를 유도하는데 효과적이고, 패모로부터 분리한 가용성 추출물은 세포분화를 촉진시키는데 효과적임을 알아낼 수 있었다. 더 나아가, 상기와 같은 효능이 있는 백지와 패모를 혼합하여 지패산을 조성하고, 이의 가용성 물질을 추출한 후 인간 백혈병 세포주인 HL-60를 대상으로 암세포에 대한 억제효과를 측정한 결과, 백지와 패모 추출물 각각에 비하여 이들을 조합한 지패산 추출물이 HL-60에 대한 세포증식억제, 세포분화촉진 및 아폽토시스 촉진에 뛰어난 효과가 있음을 발견하였다. 이에 본 발명자들은 이러한 지패산 추출물을 암 치료제 또는 치료 보조제의 유효성분으로서 활용할 수 있음에 이르러 본 발명을 완성하였다.As such demands increase, as a result of continuous research to develop a substance having the characteristics of inducing apoptosis of cancer cells, inhibiting cell proliferation and promoting cell differentiation, the present inventors have found that the soluble extract isolated from white paper It was found that it is effective in inducing apoptosis of cells, and that the soluble extract isolated from the hairs is effective in promoting cell differentiation. Furthermore, by combining the white paper and femo with the above-mentioned efficacy to form a jipasan, and extracting the soluble substance thereof, after measuring the inhibitory effect on cancer cells in human leukemia cell line HL-60, white paper and femdom Compared to each of the extracts, it was found that the Zipipic acid extracts in combination thereof had an excellent effect on cell proliferation inhibition, cell differentiation and apoptosis promotion against HL-60. Accordingly, the present inventors have completed the present invention by using the zipacic acid extract can be utilized as an active ingredient of a cancer therapeutic agent or a therapeutic aid.
따라서, 본 발명의 목적은 백지와 패모를 혼합하여 조성한 지패산의 가용성 추출물을 함유함을 특징으로 하는 암 치료용 또는 치료 보조용 조성물을 제공하는데 있다.Accordingly, it is an object of the present invention to provide a composition for treating or assisting cancer, characterized in that it contains a soluble extract of Zipacic acid prepared by mixing white paper and femo.
도 1은 본 발명에 따른 지패산 추출물의 처리 농도와 시간의 경과에 따른 인간 백혈병 세포주(Human Leukemia Cells) HL-60의 세포증식억제를 측정한 결과이다.1 is a result of measuring the cell proliferation inhibition of human leukemia cell lines (Human Leukemia Cells) HL-60 over the treatment concentration and time course of the extract of Zipacic acid according to the present invention.
도 2는 백지, 패모 또는 지패산 추출물을 10㎍/㎖, 50㎍/㎖, 100㎍/㎖의 각 농도로 처리한 후 인간 백혈병 세포주 HL-60에 대한 세포증식억제를 측정한 결과이다.Figure 2 is the result of measuring the cell proliferation inhibition of the human leukemia cell line HL-60 after treatment with white paper, femo or Zipanoic acid extract at each concentration of 10 μg / ml, 50 μg / ml, 100 μg / ml.
도 3은 인간 백혈병 세포주 HL-60에 백지, 패모 또는 지패산 추출물을 100㎍/㎖의 농도로 각각 처리하고 48시간 후 세포분화율을 측정한 결과이다. 여기에서, 도 3A 및 도 3B는 약물이 처리되지 않은 대조군 세포 및 지패산 추출물 100㎍/㎖이 처리된 실험군을 각각 광학현미경으로 관찰한 결과이고, 도 3C는 백지, 패모 또는 지패산 추출물을 100㎍/㎖의 농도로 각각 처리한 후 세포분화율을 비교한 결과이다.Figure 3 is the result of measuring the cell differentiation rate 48 hours after the white leukemia, femo or Zipacic acid extract to the human leukemia cell line HL-60 at a concentration of 100㎍ / ㎖ respectively. 3A and 3B are the results of observing the experimental group treated with 100 μg / ml of the control cells and the zipamic acid extract, which were not treated with the drug, respectively, and FIG. After treatment with the concentration of ㎍ / ㎖ each cell differentiation rate is a result.
도 4는 인간 백혈병 세포주 HL-60에 백지, 패모 또는 지패산 추출물을 100㎍/㎖의 농도로 각각 처리하고 아폽토시스율을 측정한 결과이다. 여기에서, 도 4A 및 도 4B는 약물이 처리되지 않은 대조군 세포 및 백지 추출물 100㎍/㎖이 처리된 세포를 각각 DAPI 핵염색하고 형광현미경으로 관찰한 결과이고, 도 4C는 백지, 패모 또는 지패산 추출물을 100㎍/㎖의 농도로 각각 처리한 후 아폽토시스율을 비교한 결과이다.Figure 4 shows the results of treatment of white leukemia, famo or jipasan extracts in human leukemia cell line HL-60 at a concentration of 100 µg / ml and the apoptosis rate. Here, Figures 4A and 4B are the results of observing the control cells untreated with the drug and cells treated with 100 μg / ml of white paper extract, respectively, by DAPI nucleostaining and fluorescence microscopy, and FIG. 4C shows white paper, femo or Zipacic acid. The results of the treatment of the extracts at a concentration of 100 μg / ml and the apoptosis rate were compared.
상기한 목적을 달성하기 위하여, 본 발명은 백지와 패모를 혼합하여 조성한 지패산의 가용성 추출물이 암세포, 특히 인간 백혈병 세포주 HL-60의 세포증식을 억제시키고 아폽토시스와 분화를 촉진시키는 뛰어난 효능이 있음을 제공함을 특징으로 한다. 이를 통하여, 상기 지패산 가용성 추출물을 함유함으로써 암, 특히 백혈병에 대한 치료 효능이 있는 조성물을 제공함을 특징으로 한다.In order to achieve the above object, the present invention is that the soluble extract of Zipacic acid, which is a mixture of white paper and femo, has an excellent effect of inhibiting cell proliferation and promoting apoptosis and differentiation of cancer cells, especially human leukemia cell line HL-60. It is characterized by providing. Through this, it is characterized by providing a composition having a therapeutic effect against cancer, in particular leukemia, by containing the zipa soluble extract.
이하, 본 발명을 더욱 구체적으로 설명한다.Hereinafter, the present invention will be described in more detail.
지패산(芷貝散)은 백지와 패모가 동등하게 구성된 처방으로, 전통적으로 우리나라를 비롯한 동양에서 거풍소증, 해열 및 청열 해소 등에 사용되어 왔다(윤용갑, 東醫方劑와 處方解說, 醫聖堂, 서울, p. 63, 65, 1998; 허준, 東醫寶鑑, 법인문화사, 서울, p. 690-692, 1409, 1475, 1999).Jiphaesan (芷貝 散) is a prescription composed of white paper and a feather, and has been traditionally used for relieving epidemics, fever and clarity in the East including Korea (Yun Yong-gap, 東 醫 方劑 and 處方 解說, 醫 聖堂, Seoul) , p. 63, 65, 1998; Heo Jun, 東 醫 寶 鑑, Corporate Culture History, Seoul, p. 690-692, 1409, 1475, 1999).
백지(白芷,Angelica Dahurica)는 다년생 초본인 구릿대 및 개구릿대의 뿌리로, 祛風解表, 消腫止痛, 通鼻止帶의 효능이 있어(신민교, 臨床本草學, 永林社, 서울, p. 506, 636, 1996) 두통, 치통, 비염 등을 치료한다고 하였으며(신민교, 本草求眞, 木과土, 서울, pp. 245-246, 1999; 신민교 등, 鄕藥生藥大事典, 永林社, 서울, pp. 170-171, 409-411, 1990), 약리적으로는 해열, 진통작용과 항균작용이 있는 것으로 밝혀져 있다(신민교, 本草求眞, 木과土, 서울, pp. 245-246, 1999; 신민교 등, 鄕藥生藥大事典, 永林社, 서울, pp. 170-171, 409-411, 1990). Angelica Dahurica is the root of perennial herbaceous and ulcerous, and has the effect of 효능 風 解 表, 消腫 止痛, 通 鼻 止 帶 (Shinminkyo, 臨床 本草 學, 永 林 社, Seoul, p 506, 636, 1996) Headache, Toothache, Rhinitis, etc. (Shinmingyo, 本草 求 眞, 木, 土, Seoul, pp. 245-246, 1999; Shinmingyo, etc.) , Seoul. 1999; Shin Min-kyo et al., Yeong-Shin-Dae-Jeong, Yonglin, Seoul, pp. 170-171, 409-411, 1990).
패모(貝母,Fritillariae Verticillata)는 백합과에 속하는 다년생 초본인 조선패모, 부전패모 기타 패모속 식물의 비늘줄기를 말린 것으로, 폐에 작용하여 열을 내리고, 담을 삭이며 폐를 보하여 기침을 멈추고 가래를 없애는 작용이 있다(신민교, 臨床本草學, 永林社, 서울, p. 506, 636, 1996). Fritillariae Verticillata is a dried perennial herb of Korean perennial herbaceous, dysfunctional and other family plants, which act on the lungs to reduce heat, cut off the wall, protect the lungs, stop coughing and sputum It has the effect of eliminating the problem (Shinmin-kyo, Shin-gyo-kyo, Yonglin, Seoul, p. 506, 636, 1996).
한편, 백지와 패모는 모두 항암작용이 있는 것으로 보고된 바 있으며, 특히 패모는 자궁경부암, 폐암, 유방암, 림프종 등에 효과가 있는 것으로 알려져 있으나 그 작용기전은 밝혀지지 않고 있는 실정이다(李宇彬 등, 抗癌中藥藥理與應用, 黑龍江科學技術出版社, 哈爾濱, pp. 194-195, 444-445, 1999; 苗明三 등, 法定中藥藥理與臨床, 世界圖書出版公司, 西安, p. 339, 1998).On the other hand, both white paper and femdom has been reported to have an anti-cancer effect, in particular, the femoral cervical cancer, lung cancer, breast cancer, lymphoma is known to be effective, but the mechanism of action is not known (李 宇 彬, 抗) 94 中藥 藥理 與 應用, 黑龍江 科學 技術 出版社, 哈爾濱, pp. 194-195, 444-445, 1999; .
본 발명에서는 백지의 가용성 추출물이 암세포에서 당업계에 통상적으로 주지되어 있는 아폽토시스의 특징들, 예를 들면 (1) 염색질과 핵분열을 포함한 전형적인 아폽토시스 세포의 형태학적 변화; (2) 세포주기상 서브 G1기로의 전이 활성화 등을 현저하게 유발할 수 있음을 밝혔다. 또한 패모의 가용성 추출물이 암세포의 분화를 유도하는 효과가 있음을 증명하였다.In the present invention, the soluble extract of white paper is characterized by apoptosis characteristics commonly known in the art in cancer cells, such as (1) morphological changes in typical apoptotic cells, including chromatin and fission; (2) it was found that it can significantly induce the activation of metastasis to the sub G 1 phase in the cell cycle. It was also demonstrated that the soluble extracts of the hairs have the effect of inducing the differentiation of cancer cells.
본 발명에 따른 지패산은 백지와 패모를 일정 비율로 혼합하여 조성한 것이다. 이때, 백지와 패모를 동량 혼합하는 것이 바람직하나 이에 한정하는 것은 아니다.Zipacic acid according to the present invention is a mixture of white paper and famo at a certain ratio. At this time, it is preferable to mix the same amount of white paper and feather, but is not limited thereto.
본 발명에 따른 지패산의 가용성 추출물은 지패산을 물을 용매로 하여 열수추출한 다음 그 상층액을 동결건조하여 제조한 것을 이용하는 것이 바람직하다. 그러나, 용매, 추출방법 또는 제조방법은 이에 한정하는 것은 아니고, 공지의 것들을 적의 선정하여 목적에 맞게 사용할 수 있다.Soluble extract of Zipacic acid according to the present invention, it is preferable to use that prepared by hot-water extraction using Zipacic acid as a solvent and then lyophilized the supernatant. However, a solvent, an extraction method, or a manufacturing method is not limited to this, A well-known thing can be selected suitably and can be used according to the objective.
본 발명에 따른 지패산의 가용성 추출물은 암세포의 성장과 분화를 촉진하는 효과가 뛰어나고, 특히 백지와 패모를 각각 구분하여 적용한 결과에 비하여 항암 효과가 더욱 뛰어난 특징이 있다. 보다 구체적으로는 본 발명에 따른 지패산의 가용성 추출물은 암세포, 특히 인간 급성백혈병 세포인 HL-60의 세포증식을 억제하고, 세포분화 및 아폽토시스를 촉진하는 효과가 매우 뛰어나다.Soluble extract of Zipacic acid according to the present invention has an excellent effect of promoting the growth and differentiation of cancer cells, in particular, the anti-cancer effect is more excellent than the result of separately applying white paper and famo. More specifically, the soluble extract of Zipacic acid according to the present invention is very effective in inhibiting cell proliferation of cancer cells, especially human acute leukemia cells, HL-60, and promoting cell differentiation and apoptosis.
따라서 본 발명에 따른 지패산의 가용성 추출물은 인간 백혈병 뿐만 아니라 각종의 암을 치료 또는 치료 보조하는데 있어 유용하게 사용될 수 있다. 지패산 추출물을 유효성분으로 함유하는 조성물은 상기와 같이 각종 암세포들의 아폽토시스는 물론 세포분화를 촉진함으로써 암세포의 증식을 효과적으로 차단하는 효과가 뛰어나므로, 백혈병, 유방암, 간암, 자궁경부암, 위암, 난소암, 방광암, 폐암, 상피종암 등의 각종 암, 더욱 바람직하게는 백혈병의 치료에 유용하게 투여될 수 있다.Therefore, the soluble extract of Zipacic acid according to the present invention can be usefully used for treating or assisting various cancers as well as human leukemia. As the composition containing the zipaic acid extract as an active ingredient, as described above, it is excellent in blocking cancer cell proliferation by promoting apoptosis of various cancer cells as well as cell differentiation, thus, leukemia, breast cancer, liver cancer, cervical cancer, stomach cancer, ovarian cancer , Bladder cancer, lung cancer, epithelial cancer, and various cancers, more preferably can be administered in the treatment of leukemia.
이러한 관점에서 본 발명에 따른 아폽토시스 유도 및 세포분화제 조성물은 암세포, 바람직하게는 백혈병 세포에서 아폽토시스를 활성화시키고, 세포분화를 촉진함으로써 암치료, 바람직하게는 백혈병 치료에 유용하게 이용될 수 있지만, 본원 발명의 조성물을 적용할 수 있는 질환은 이에 한정되지 않고, 당업계에 통상적으로 주지되어 있는 아폽토시스의 유도와 세포분화를 통한 세포 증식 억제와 관련된 질환들의 치료를 위해서도 유용하게 사용될 수 있다.In this respect, the apoptosis induction and cell differentiator composition according to the present invention can be usefully used for cancer treatment, preferably leukemia treatment by activating apoptosis and promoting cell differentiation in cancer cells, preferably leukemia cells. The disease to which the composition of the present invention can be applied is not limited thereto, and may be usefully used for the treatment of diseases related to inhibition of cell proliferation through induction of apoptosis and cell differentiation, which are commonly known in the art.
또한, 본 발명에 따른 지패산 추출물을 함유하는 조성물은 암 치료에 적용할 경우 그 자체를 단독으로 투여하는 것도 가능하지만 다른 암치료제 또는 암치료제 조성물, 약제 및 다른 여러 물리학적 또는 의학적 치료방법과 병행하여 사용함으로써 그 효과를 극대화시킬 수 있다.In addition, the composition containing the zipaic acid extract according to the present invention may be administered alone when applied to cancer treatment, but in combination with other cancer treatment or cancer treatment composition, drugs and other physical or medical treatment methods. By using it, the effect can be maximized.
본 발명에 따른 지패산 추출물을 함유하는 조성물에서 유효성분인 지패산의 함량은 투여방법, 제제형태, 환자의 나이, 환자의 체중, 환자의 감수성 및 질환의 상태에 따라 적절하게 선택되어질 수 있으며, 구체적으로 제한되는 것은 아니지만일반적으로 지패산을 0.1∼200㎎/㎏체중의 농도로 투여되도록 제형화될 수 있다. 물론, 유효성분의 투여량이 상기 범위를 벗어난 양일 수도 있다.The content of zipaic acid as an active ingredient in the composition containing zipaic acid extract according to the present invention may be appropriately selected depending on the method of administration, the type of preparation, the age of the patient, the weight of the patient, the sensitivity of the patient and the condition of the disease, In general, but not limited to, it can be formulated to administer the zipic acid at a concentration of 0.1 to 200 mg / kg body weight. Of course, the dose of the active ingredient may be an amount outside the above range.
본 발명에 따른 지패산 추출물을 함유하는 조성물은 제제로서 통상의 형태를 가질 수 있으며, 예를 들면 알약, 캅셀 형태나 드링크제, 주사제, 의약품 등의 형태로 제형화하여 사용할 수 있다. 이들은 경구 또는 각종의 비경구 투여 경로를 통하여 아폽토시스 또는 세포분화를 유도하고자 하는 부위에 투여될 수 있으며, 투여 제형에 따라 적합하고 당업자에게 주지되어 있으며 당업자가 용이하게 선정할 수 있는 각종의 부형제, 담체 또는 희석제 등을 더 함유할 수 있다.The composition containing the Zipacic acid extract according to the present invention may have a conventional form as a preparation, and may be used, for example, in the form of a pill, a capsule, a drink, an injection, a medicine, or the like. They may be administered orally or through various parenteral routes of administration to the site of induction of apoptosis or cell differentiation, and are suitable for the dosage form and are well known to those skilled in the art and can be readily selected by those skilled in the art. Or a diluent or the like.
이하, 본 발명을 실시예와 도면에 의해서 더욱 상세하게 설명하지만, 본 발명의 권리범위가 이에 의해서 한정되는 것은 아니다. 비록 하기의 실시예에서는 본 발명에 따른 조성물의 아폽토시스 유도 및 세포분화 효과에 대해서 백혈병 세포중 인간백혈병 세포 HL-60에 적용한 결과만을 보여주었지만, 본 발명의 조성물이 적용되는 범위는 상기 HL-60 세포 또는 백혈병 세포에만 적용되는 것이 아니라 각종 암세포에서도 아폽토시스와 세포분화를 유발함으로써 그 해당 암질환을 치료할 수 있다는 것은 당업자에게 자명하게 인식될 수 있을 것이다.Hereinafter, the present invention will be described in more detail with reference to examples and drawings, but the scope of the present invention is not limited thereto. Although the following examples show only the results of application to human leukemia cells HL-60 in leukemia cells for apoptosis induction and cell differentiation effects of the composition according to the present invention, the scope of the composition of the present invention is applied to the HL-60 It will be apparent to those skilled in the art that the cancer disease can be treated by inducing apoptosis and cell differentiation not only in cells or leukemia cells but also in various cancer cells.
하기의 실시예 중 실험예들은 각각의 실험들을 적어도 3번 이상 실행하여 얻어진 결과들이며, Student's t-test를 이용한 통계 분석을 이용하였다. 유의성의 한계는 p<0.01를 기준으로 정하였다.Experimental examples of the following Examples are the results obtained by performing each experiment at least three times, using a statistical analysis using Student's t-test. The limit of significance was set based on p <0.01.
[참고 실시예 1] 재료Reference Example 1 Material
하기의 시험예에서 이용되는 시약 및 시료는 다음과 같이 구입하였다.The reagents and samples used in the following test examples were purchased as follows.
RPMI-1640과 우태아혈청(fetal bovine serum; FBS)은 Gibco/BRL(Grand Island. NY. U.S.A)로부터 구입하였다. 디메틸 설폭사이드(Dimethyl sulfoxide; DMSO), 3-[4,5-디메틸티아졸-2-일]-2,5-디페닐테트라졸리움 브로마이드, 트립판 블루(trypan blue), DAPI 및 프로피디움 아이오다이드(propidium iodide; PI)는 시그마사(Sigma Chemical, St. Louis, MO, U.S.A)로부터 구입하였다. 그 외 모든 용매는 분석등급인 것을 사용하였으며 머크사(Merck, Darmstadt, Germany)로부터 구입하였다.RPMI-1640 and fetal bovine serum (FBS) were purchased from Gibco / BRL (Grand Island.NY.U.S.A). Dimethyl sulfoxide (DMSO), 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide, trypan blue, DAPI and propidium ioda Propidium iodide (PI) was purchased from Sigma Chemical, St. Louis, Mo., USA. All other solvents were of analytical grade and were purchased from Merck (Darmstadt, Germany).
[참고 실시예 2] 세포배양Reference Example 2 Cell Culture
HL-60 세포주(CCL-240)는 ATCC(American Type Culture Collection, ATCC; Rockville. MD. U.S.A)의 기탁기관으로부터 분양받았다. 상기 분양받은 HL-60 세포주를 10% 우태아혈청(FBS)이 첨가된 RPMI-1640 배양배지에서 95% 공기 및 5% 이산화탄소(CO2)가 소통되는 습기가 충분한 대기에서 37℃ 온도 조건을 유지하며 배양하였다. 세포의 지수 성장을 유지하기 위해, 2∼3일 마다 새로운 배양배지에 분주하면서 배양하였다.The HL-60 cell line (CCL-240) was distributed from the Depositary of the American Type Culture Collection (ATCC), Rockville. MD. USA. The HL-60 cell line was maintained at 37 ° C. in a humid atmosphere with 95% air and 5% carbon dioxide (CO 2 ) in RPMI-1640 culture medium supplemented with 10% fetal bovine serum (FBS). And incubated. In order to maintain exponential growth of the cells, the cells were cultured by dispensing in new culture medium every 2-3 days.
[참고 실시예 3] 지패산 가용성 추출물의 제조Reference Example 3 Preparation of Zipacic Acid Soluble Extract
백지 및 패모는 원광대학교 익산한방병원에서 구입하였다. 백지 50g과 패모 50g을 혼합하여 지패산을 제조하였다. 상기 제조된 지패산을 물 600㎖와 함께 대웅약탕기(DWP-1800P)에 넣고 2시간 30분간 100℃로 가열하여 추출하였다. 상층액을 여과한 다음 동결건조시켜 7.2g의 지패산 고형물을 얻었다.The white paper and femo were purchased from Iksan Oriental Hospital of Wonkwang University. Zipacic acid was prepared by mixing 50 g of white paper and 50 g of famo. The prepared Jipaic acid was put into Daewoong Yaktang (DWP-1800P) with 600 ml of water and extracted by heating to 100 ° C. for 2 hours 30 minutes. The supernatant was filtered and then lyophilized to yield 7.2 g of fat acid.
백지 100g과 물 600㎖, 그리고 패모 100g과 물 600㎖을 각각 상기와 같은 방법으로 추출하고 동결건조시켜 각각 12g의 백지 고형물 및 6.15g의 패모 고형물 을 얻었다. 상기 모든 고형물은 -70℃에서 보관하여 하기 시험예에 사용하였다.100 g of white paper, 600 ml of water, and 100 g of paddy and 600 ml of water were extracted and lyophilized as described above, respectively, to obtain 12 g of white paper and 6.15 g of solid, respectively. All the solids were stored at −70 ° C. and used in the following test examples.
〈시험예 1〉세포증식억제 측정Test Example 1 Cell Proliferation Inhibition
HL-60의 세포수는 트립판 블루(Trypan blue, 시그마사 구입) 염색과 헤모사이토메타(hemocytometer)를 사용하였으며, 광학현미경(짜이스사, 독일)을 이용하여 죽은 세포와 살아있는 세포를 계수하였다.The cell number of HL-60 was trypan blue (Sigma Co., Ltd.) staining and hemocytometer (hemocytometer), and the dead cells and living cells were counted by using an optical microscope (Cays, Germany).
급성백혈병은 미분화된 줄기세포가 무제한으로 증식하는 질병으로, 따라서 이를 치료하기 위해서는 백혈병 세포의 증식을 억제하는 것이 가장 효과적이다. 이에 지패산 가용성 추출물이 급성백혈병 세포인 HL-60 세포의 증식에 어떠한 영향을 주는지를 조사하였다.Acute leukemia is a disease in which undifferentiated stem cells proliferate indefinitely, and therefore, it is most effective to suppress the proliferation of leukemia cells in order to treat them. We investigated the effect of zipiic acid soluble extract on the proliferation of HL-60 cells, acute leukemia cells.
상기 참고 실시예 3에서 제조한 지패산 가용성 추출물을 10㎍/㎖, 50㎍/㎖, 100㎍/㎖의 농도로 준비한 것 및 PBS 완충액(대조군)을 각각 상기 배양한 급성백혈병 세포주 HL-60에 처리하고 시간의 경과에 따른 세포수를 계수하였다.The zipaic acid soluble extract prepared in Reference Example 3 was prepared at concentrations of 10 μg / ml, 50 μg / ml and 100 μg / ml, and PBS buffer (control), respectively, into the cultured acute leukemia cell line HL-60. The cells were treated and counted over time.
그 결과, 도 1에서와 같이 지패산 가용성 추출물의 처리 농도가 증가할수록시간의 경과에 따른 급성백혈병 세포주 HL-60의 세포증식이 대조군에 비해 현저히 억제되었음을 알 수 있다. 특히 세포 배양시간 48시간에 있어서, 지패산 가용성 추출물 100㎍/㎖ 농도에서의 세포증식은 10㎍/㎖ 또는 50㎍/㎖와 비교할 때 현저히 억제되었다.As a result, it can be seen that as the treatment concentration of the zipaic acid soluble extract increased as shown in FIG. 1, cell proliferation of the acute leukemia cell line HL-60 with time was significantly suppressed compared to the control group. Particularly at 48 hours of cell culture time, cell proliferation at 100 μg / ml concentration of zipaic acid soluble extract was significantly inhibited compared to 10 μg / ml or 50 μg / ml.
한편 백지 추출물, 패모 추출물 또는 지패산 추출물을 각각 구분하여 10㎍/㎖, 50㎍/㎖, 100㎍/㎖의 농도로 HL-60 세포에 처리하고 48시간 후에 세포수를 측정함으로써 세포증식에 미치는 이들 약물의 효과를 비교하였다.On the other hand, white paper extracts, femo extracts, or Zipacic acid extracts were separately treated to HL-60 cells at concentrations of 10 µg / ml, 50 µg / ml and 100 µg / ml, and the cell number was measured after 48 hours. The effects of these drugs were compared.
그 결과, 대조군에 비해서 백지 추출물, 패모 추출물 및 지패산 추출물 모두에 있어서 HL-60 세포의 증식억제효과가 뚜렷히 나타났다(도 2). 특히, 같은 농도의 백지, 패모 또는 지패산 가용성 추출물의 세포증식 억제능력을 비교한 결과 모든 농도에 있어서 지패산 추출물의 HL-60 세포증식 억제효과가 백지 또는 패모 추출물의 그것에 비해서 뛰어난 것으로 나타났다(p<0.01).As a result, the proliferation inhibitory effect of HL-60 cells was clearly seen in all of the white paper extract, femo extract and Zipacic acid extract compared to the control (Fig. 2). In particular, the comparison of cell proliferation inhibitory ability of baekchi, femo or Zipanoic acid soluble extracts of the same concentration showed that HL-60 cell proliferation inhibitory effect of Zipacic acid extract was superior to that of white paper or femo extract at all concentrations (p <0.01).
이를 통하여, 급성백혈병 세포의 증식을 효과적으로 억제시키기 위해서는 백지와 패모를 단독으로 사용하는 것보다 동시에 조성한 지패산의 사용이 매우 유용함을 알 수 있었다.Through this, it was found that in order to effectively inhibit the proliferation of acute leukemia cells, the use of Zipacic acid prepared at the same time is more useful than the use of white paper and femo alone.
〈시험예 2〉세포분화율 측정<Test Example 2> Measurement of cell differentiation rate
지패산 가용성 추출물 100㎍/㎖ 및 PBS 완충액(대조군)을 각각 48시간동안 처리한 급성백혈병 세포 HL-60의 세포분화상태를 관찰하기 위하여, 국제진단시약회사(國際診斷試藥 社, 일본)의 제품인 Wright-Giemsa kit를 사용하여 세포 염색을 실시하였다. 먼저, 세포를 슬라이드 글라스에 사이토스핀(Cytospin)을 사용하여 부착시키고 1차 고정액으로 5분간 고정한 다음 2차 Giemsa 핵염색시약으로 3분간 핵을 염색하였다. 마지막으로 5분간 3차 염색액으로 세포질을 염색하고 PBS로 3회 반복 세척한 후 광학현미경하에서 ×1000의 배율로 관찰하였다.In order to observe the cellular differentiation of acute leukemia cells HL-60 treated with 100 µg / ml of zipamic acid soluble extract and PBS buffer (control) for 48 hours, respectively, the International Diagnostic Reagent Co., Ltd. (Japan) Cell staining was performed using the product Wright-Giemsa kit. First, the cells were attached to the slide glass using Cytospin, fixed for 5 minutes with the primary fixer, and stained for 3 minutes with the secondary Giemsa nuclear staining reagent. Finally, the cytoplasm was stained with a tertiary dye solution for 5 minutes, washed three times with PBS, and observed at a magnification of × 1000 under an optical microscope.
그 결과, 대조군(도 3A)에 비해 지패산 가용성 추출물 처리군(도 3B)에서는 형태적으로 분화된 세포의 수가 많이 증가되었으며, 증식하는 세포는 현저히 줄어들었음을 확인할 수 있었다.As a result, it was confirmed that the number of morphologically differentiated cells increased significantly in the treatment group (FIG. 3B) of zipamic acid soluble extract compared to the control group (FIG. 3A), and the proliferating cells were significantly reduced.
지패산 가용성 추출물이 HL-60 세포의 분화를 유도하는지를 다음과 같이 확인하였다. 과립백혈구(granulocyte)/단핵구(monocyte)/대식세포(macrophage)를 구분하는 표지로 FTTC 형광물질이 부착된 CD11b에 대한 항체를 이용하였다. HL-60 세포를 24-웰 배양 플라스크에 웰당 1×105이 되도록 준비한 뒤에, 100㎍/㎖의 백지, 패모 또는 지패산 추출물을 처리하고 48시간 동안 배양하였다. 상기 추출물을 처리하여 배양한 세포를 500×g에서 원심분리하여 수거한 후에 0.5% BSA와 0.1% 아지드화 나트륨(sodium azide)을 포함한 인산완충액(pH 7.2)으로 3회 세척하였다. 그 다음, 200㎕의 인산완충액에 1×106의 세포를 넣고 FITC가 결합된 CD11b 항체를 25㎍/㎖이 되도록 넣은 뒤 빛을 차단한 상태로 4℃에서 30분간 반응시켰다. 반응이 끝난 후 반응하지 않은 항체를 제거하기 위하여 0.5% BSA와 0.1% 아지드화 나트륨을 포함한 인산완충액으로 3회 원심세척한 뒤에 FACS vantage(Becton Dickinson, 미국)을 사용하여 세포지면에 발현된 각각의 항체를 측정하였다.It was confirmed as follows whether the zipamic acid soluble extract induces differentiation of HL-60 cells. Antibodies against CD11b with FTTC fluorescent material were used as markers to distinguish granulocyte / monocyte / macrophage. HL-60 cells were prepared in a 24-well culture flask at 1 × 10 5 per well, then treated with 100 μg / ml of white paper, femo, or zipic acid extracts and incubated for 48 hours. The cells treated with the extracts were harvested by centrifugation at 500 × g and washed three times with phosphate buffer (pH 7.2) containing 0.5% BSA and 0.1% sodium azide. Then, 1 × 10 6 cells were added to 200 μl phosphate buffer solution, and the FITC-coupled CD11b antibody was added to 25 μg / ml and reacted at 4 ° C. for 30 minutes while blocking light. After completion of the reaction, centrifuged three times with phosphate buffer containing 0.5% BSA and 0.1% sodium azide to remove unreacted antibodies, and then expressed on the cell surface using FACS vantage (Becton Dickinson, USA). The antibody of was measured.
세포분화율 측정에 관한 상기 실험 결과를 도 3C에서와 같이 백분율로 나타내었다. 백지는 대조군과 거의 유사한 정도로 세포분화에 효과가 없었지만, 패모 처리군의 경우 40% 이상 세포가 분화되는 우수한 효과를 보였다. 특히 지패산 처리군에서는 세포분화율이 60% 이상으로 나타났는데, 이는 지패산이 HL-60 세포의 분화를 촉진시키는 뛰어난 효과가 있음을 보이는 것이다.The results of the above experiments on the determination of cell differentiation are expressed in percentage as in FIG. 3C. The white paper had no effect on cell differentiation to the same extent as the control group, but showed a good effect of more than 40% of the cells in the famo treated group. In particular, the cell differentiation rate was 60% or more in the treatment group of Zipisan, indicating that Zipisan has an excellent effect of promoting the differentiation of HL-60 cells.
결과적으로, 급성백혈병 세포의 증식을 억제하고 분화를 촉진시키기 위해서는 패모를 단독으로 사용할 수도 있지만, 그 효과를 극대화시키기 위해서는 패모와 백지를 혼합한 지패산을 사용하는 것이 바람직함을 알 수 있다.As a result, the hair loss may be used alone to suppress the proliferation and promote differentiation of acute leukemia cells, but it can be seen that it is preferable to use a Zipacic acid mixed with the hair and white paper to maximize the effect.
〈시험예 3〉DAPI를 이용한 핵 염색<Test Example 3> Nuclei staining using DAPI
HL-60 세포(1×106cells/㎖)를 10%의 우태아혈청(fetal bovine serum)이 포함된 RPMI 1640 배지에 백지 100㎍/㎖를 첨가하거나 첨가하지 않은 상태에서 6웰 접시(6-well dishes)에서 4시간 동안 배양하였다. 배양 후, 1㎖의 세포 부유액을 1,000rpm에서 3분 동안 원심분리하여 세포를 침전시킨 다음, 세포 침전물에 4%의 중성 완충된 100㎕의 파라포르말린을 첨가하여 고정시켰다. 고정된 50㎕의 세포 부유액을 슬라이드에 옮기고, 실온에서 건조시켰다. 고정된 세포를 인산완충액으로 세척한 후 건조시키고, DNA-특이적 플루오로크롬(fluorochrom) DAPI (4,6-Diamidino-2-phenylindole)으로 1분간 염색하였다. 부착된 세포를 인산완충액으로 세척한 후 건조시키고, 90% 글리세롤(glycerol)로 봉입하였다. 완성된 슬라이드를 형광현미경을 이용해 관찰하였다. 그 결과를 도 4A 및 4B에 나타내었다.HL-60 cells (1 × 10 6 cells / ml) in RPMI 1640 medium containing 10% fetal bovine serum, with or without 100 μg / ml of white paper -well dishes) for 4 hours. After incubation, 1 ml of the cell suspension was centrifuged at 1,000 rpm for 3 minutes to precipitate the cells, and then fixed to the cell precipitate by adding 4% neutral buffered 100 µl of paraformalin. Immobilized 50 μl of cell suspension was transferred to slides and dried at room temperature. Immobilized cells were washed with phosphate buffer, dried and stained with DNA-specific fluorochrom DAPI (4,6-Diamidino-2-phenylindole) for 1 minute. The attached cells were washed with phosphate buffer, dried and encapsulated with 90% glycerol. The completed slide was observed using a fluorescence microscope. The results are shown in FIGS. 4A and 4B.
도 4A 및 4B에서 보는 바와 같이, 백지는 아폽토시스를 통한 세포 사멸의 전형적인 형태학적 특징으로서 핵염색질의 응축, 핵절편화 및 아폽토시스 바디 등을 유발시켰음을 확인하였다.As shown in Figures 4A and 4B, it was confirmed that white paper caused condensation, nucleation and apoptosis body of nuclear chromosome as typical morphological features of cell death through apoptosis.
또한 상기의 방법과 동일한 방법으로 패모 또는 지패산 추출물 100㎍/㎖을 HL-60 세포에 처리한 후 DAPI 염색을 하고 그 결과로써 HL-60 세포의 아폽토시스율을 백분율로 나타내었다(도 4C). 예상한 바와 같이 백지의 경우 아폽토시스율이 현저히 높은 반면, 패모의 경우는 대조군과 거의 차이가 없었고, 이들 약물을 조합한 지패산의 경우는 백지의 영향으로 아폽프토시스율이 대조군에 비해 현저하였으나, 백지의 경우보다는 약간 낮게 나타났다. 이상의 결과는 상기 시험예 2에서 밝힌 바와 같이, 패모가 HL-60 세포의 분화를 촉진시켰기 때문에 이러한 패모의 영향을 받아 지패산 처리군이 백지 처리군보다 아폽토시스율이 낮게 나타난 것이라 사료된다.In addition, the same method as the above method was treated with 100 μg / ml of the extract of Famo or Zipanic acid to HL-60 cells, and then stained with DAPI. As a result, the apoptosis rate of HL-60 cells was expressed as a percentage (FIG. 4C). As expected, the apoptosis rate was significantly higher in the case of white paper, while the apoptosis rate was significantly higher in the case of femo, and in the case of Zipacic acid with these drugs, the apoptosis rate was significantly higher than that of the control group. It was slightly lower than the blank paper. As shown in Test Example 2 above, it was thought that the apoptosis treatment group showed a lower apoptosis rate than the white paper treatment group due to the influence of the hair follicles because the hair follicles promoted differentiation of HL-60 cells.
이상에서 살펴본 바와 같이, 백지와 패모를 혼합하여 조성한 지패산으로부터 추출한 가용성 추출물은 암세포, 특히 급성백혈병 세포의 증식을 억제할 뿐만 아니라, 세포분화 및 아폽토시스를 촉진하는 활성이 매우 뛰어난 효과가 있으므로, 각종 암, 특히 백혈병을 치료하기 위한 항암제 조성물의 유효성분으로 유용하게 이용될 수 있다.As described above, the soluble extract extracted from Zipacic acid, which is a mixture of white paper and femo, not only inhibits the proliferation of cancer cells, especially acute leukemia cells, but also has an excellent effect of promoting cell differentiation and apoptosis. It can be usefully used as an active ingredient of an anticancer agent for treating cancer, especially leukemia.
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