WO2012030150A2 - Composition contenant un extrait de solvant organique d'ailante glanduleux pour prévenir et traiter le cancer de la prostate - Google Patents

Composition contenant un extrait de solvant organique d'ailante glanduleux pour prévenir et traiter le cancer de la prostate Download PDF

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Publication number
WO2012030150A2
WO2012030150A2 PCT/KR2011/006421 KR2011006421W WO2012030150A2 WO 2012030150 A2 WO2012030150 A2 WO 2012030150A2 KR 2011006421 W KR2011006421 W KR 2011006421W WO 2012030150 A2 WO2012030150 A2 WO 2012030150A2
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prostate cancer
extract
composition
preventing
cells
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PCT/KR2011/006421
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English (en)
Korean (ko)
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WO2012030150A3 (fr
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황성연
정경채
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주식회사 한국전통의학연구소
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Publication of WO2012030150A2 publication Critical patent/WO2012030150A2/fr
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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)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to a novel use of low myrtle skin extract. Specifically, the present invention relates to a therapeutic composition containing as an active ingredient low myrtle bark extract exhibiting excellent prophylactic or therapeutic efficacy against prostate cancer.
  • Prostate cancer is a malignant tumor that occurs in the prostate gland, and is one of the most common cancers among men in the West, and it is the fastest growing cancer in men due to recent dietary habits in Korea. do. In developed countries such as North America and Western Europe, it is the most common cancer that accounts for about 20% of male cancers, and the United States has the highest frequency of male cancers occurring annually, and ranks second after lung cancer among cancer deaths. In Korea, the incidence of prostate cancer has increased significantly in recent years due to an increase in life expectancy, an increase in the elderly, westernization of a diet, development of diagnostic technology, and increased awareness of prostate cancer.
  • Prostate cancer is known to be caused by hormones, dietary habits, and chemicals in addition to genetic predisposition.
  • prostate cancers are cancers of gland cells in the prostate, which spread well to lymph nodes and bones. About 90% of prostate cancers are proliferated by male hormones that are produced in your body. For this reason, hormonal therapies that inhibit cancer growth and kill some of the cancer cells by inhibiting the action of male hormones are most commonly used in the treatment of prostate cancer (Greenlee, R. T et al., Cancer statistics.CA Cancer J. Clin 50: 7 (2000).
  • Treatments for prostate cancer include (a) radical prostatectomy (b) radiation (c) chemotherapy (d) hormonal therapy.
  • hormonal therapy is a so-called androgen removal method that inhibits male hormone secretion or blocks hormone production by surgery, since a significant portion of prostate cancer cells multiply androgen-dependently. This method has a temporary therapeutic effect in more than 80% of patients, such as cancer suppression or lesion reduction.
  • HRPC hormone-refractory prostate cancer
  • conventional anticancer drugs, chemotherapy, or radiation therapy do not show a great effect, so a new treatment method is required (Fong, L. et al., Induction of tissue-specific autoimmune prostatitis with prostatic acid phosphatase immunization: implications for immunotherapy of prostate cancer.J. Immunol. 159: 3113. (1997).
  • Ailanthi cortex (Ailanthic Radicis Cortex) is a broad-leaved arborescent of the Simaroubaceae tree, which is obtained by collecting root or stem bark of Ailanthus altissima SWINGLE.
  • the chemical components are phenolic substances, including 3,4,5-trimethoxyphenol, p-comaric acid, vanillin, vanillic acid, coniferyl aldehyde, 2 , 6-dimethyxybenzoquinone, trans-triacontyl-4-hydroxy-3-methoxycinnamate have been identified, and flavonoid compounds 5,7-dihydroxychromone-7-neohesperidoside and naringin have been reported, and mesosin, tannin, and flovaphene in the root bark have been reported.
  • 1- (1 ', 2'-dihydroxyethyl) -4-methoxy-beta-carboline (1- (1', 2'-Dihydroxyethyl)-4-methoxy-beta-carboline), 1-meth 1-Methoxycanthin-6-one-3-N-oxide, Ailanthinone, Ailanthone, Amarolide-11-Acetate -11-acetate, Canthin-6-one, Chaparrin, Chaparrinone, Chaparrolide, Quassin, Quecitrin, Shinjudilactone, Shinjuglucoside A, B, C, D, E, F, G and the like are known.
  • low-muscle white skin The pharmacological activity of low-muscle white skin was HIV-1 inhibitory activity [Jang, Young-Soo et al., 2003, Korean Journal of Pharmacognosy, 24: 28-32], anticancer activity [Kim Jong et al., 1997, Korean Journal of Pharmacognosy, 28 (1): 54-58] , Anti-inflammatory, diuretic, antipyretic [Lee Cheol-won et al., 1986, East-West Medical 11 (2): 30-47].
  • low root baekpi is the horse's skin from the roots of the leather, it is effective for the symptom that blood is mixed in the stool, diarrhea and stool.
  • the present inventors completed the present invention by confirming that the myofi myopia extract can effectively kill prostate cancer cells during the study of myopic myocardium.
  • the present invention provides a composition for the prevention and treatment of prostate cancer, containing the organic solvent extract of the low- necked cortex Ailanthi cortex as an active ingredient.
  • the organic solvent is preferably ethanol, wherein the ethanol extract is more preferably extracted at 50 ° C. for 24 hours or dried and concentrated at 45 ° C. under reduced pressure, and also Most preferably, the ethanol is 95%.
  • the composition of the present invention comprises a low myrtle skin extract as an active ingredient, and may further include a pharmaceutically acceptable carrier or diluent.
  • the extract of the present invention is possible at any part of the myofi myopia, but preferably is extracted from the root bark.
  • the extraction solution may be obtained by extraction with water or an organic solvent, and examples of the organic solvent may include lower alcohols, acetone, chloroform, methylene chloride, ether, ethyl acetate, and hexane.
  • Lower alcohols include methanol, ethanol, propanol and butanol, with ethanol being most preferred.
  • the ethanol extract may be prepared by filtration.
  • the filtrate obtained by filtering the extract may be concentrated under reduced pressure.
  • the extraction process may be repeated two or more times as necessary, and the extract obtained after filtration may be lyophilized or dried under reduced pressure to obtain a powder form.
  • the anticancer composition of the present invention includes low myofipi as an active ingredient, and may further include a pharmaceutically acceptable carrier or diluent.
  • a pharmaceutically acceptable carrier is a pharmaceutically acceptable substance such as a liquid or solid filler, diluent, excipient or solvent which serves to transport the active ingredient from one organ or part of the body to another organ or part of the body. , Composition or vehicle.
  • the composition of the present invention alone or in radiation therapy or chemotherapy (cell growth arrest or cytotoxic substance, antibiotic-type substance, alkylating agent, anti-metabolic substance, hormone, immuno-agent, interferon type Substances, cyclooxygenase inhibitors (e.g.
  • COX-2 inhibitors COX-2 inhibitors
  • metallomatrix protease inhibitors metallomatrix protease inhibitors
  • teromerase inhibitors tyrosine kinase inhibitors
  • anti-growth factor receptor materials anti-HER substances
  • anti-EGFR substances anti-EGFR substances
  • anti Other anticancer agents such as angiogenesis agents, farnesyl transferase inhibitors, ras-raf signal conduction pathway inhibitors, cell cycle inhibitors, other cdk inhibitors, tubulin binders, topoisomerase I inhibitors, topoisomerase II inhibitors, etc. It can be administered in combination with treatment.
  • compositions of the present invention may include one or more chemotherapeutic agents optionally contained in liposome formulations (e.g., taxanes, taxane derivatives, encapsulated taxanes, CPT-11, camptothecin derivatives, anthracycline glycosides, doxorubicin, idarubicin).
  • chemotherapeutic agents optionally contained in liposome formulations (e.g., taxanes, taxane derivatives, encapsulated taxanes, CPT-11, camptothecin derivatives, anthracycline glycosides, doxorubicin, idarubicin).
  • liposome formulations e.g., taxanes, taxane derivatives, encapsulated taxanes, CPT-11, camptothecin derivatives, anthracycline glycosides, doxorubicin, idarubicin.
  • compositions of the invention When formulated at a constant dose, such combinations use the compositions of the invention within the dosage ranges described below and other pharmaceutically active substances within the approved dosage ranges.
  • the compositions of the present invention can be used sequentially with known anticancer agents when the combination formulation is inappropriate.
  • the effective daily dose of the active ingredient in the anticancer composition of the present invention may be determined according to the age, sex, site of application, frequency of administration, administration time, formulation, type of adjuvant, and the like.
  • composition of the present invention can be administered via a route commonly used for anticancer treatment.
  • the anticancer composition of the present invention may be prepared in various formulations depending on the dosage form.
  • it may be administered in oral form of tablets, capsules, dragees or film encapsulated tablets, solutions or suspensions, parenteral forms of intramuscular, intravenous and / or intrathecal and / or intrathecal injection or infusion.
  • oral solids may be used in combination with the active compound in diluents (e.g. lactose, dextrose, sucrose, cellulose, corn starch or potato starch), lubricants (e.g.
  • silica, talc, stearic acid, Magnesium or calcium stearate and / or polyethylene glycol binders (e.g. starch, arabic gum, gelatin methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone), disintegrants (e.g. starch, alginic acid, Alginate or sodium starch glycolate), formal mixtures, dyes, sweeteners, wetting agents (eg lecithin, polysorbates, laurylsulfate) and pharmacologically inert substances generally used in pharmaceuticals.
  • binders e.g. starch, arabic gum, gelatin methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone
  • disintegrants e.g. starch, alginic acid, Alginate or sodium starch glycolate
  • formal mixtures dyes
  • sweeteners e.g lecithin, polysorbates, laurylsulfate
  • wetting agents eg lec
  • Liquid dispersions for oral administration can be, for example, syrups, emulsions and suspensions.
  • Suspensions and emulsions may contain, for example, natural gums, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose or polyvinyl alcohol as carriers.
  • Suspensions or solutions for intramuscular injection, together with the active compound may be used as a pharmaceutically acceptable carrier, e.g. sterile water, olive oil, ethyl oleate, glycols (e. G. Propylene glycol) and, if necessary, in suitable amounts of lidocaine hydrochloride. It may contain.
  • Solutions for intravenous injection or infusion may, for example, contain sterile water or preferably in the form of sterile, aqueous, isotonic saline solutions or carrier propylene glycol.
  • the ethanol extract of the myofibrillus pelvis kills PC-3 or DU 145 prostate cancer cells 62% and 85% at 100 ⁇ g / ml, respectively.
  • the above results demonstrate that the low myrrhizal extract of the present invention has excellent killing activity of PC-3 or DU 145 prostate cancer cells and further has prostate cancer treatment and prophylactic activity.
  • composition of the present invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level refers to a patient's sexually transmitted disease, age, severity, and drug activity.
  • the compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. It may be single or multiple doses.
  • the method of administering the composition comprising the extract or compound prepared according to the preparation method of the present invention is preferably oral administration or intravenous administration.
  • the effective dose is oral administration, it is usually 1 to 1 time per adult. 500 mg / kg is preferred, and in the case of intravenous administration, 1 to 100 mg / kg is preferred, and may be administered 2-3 times a day.
  • Dosage levels for a particular patient may vary depending on gender, age, myofascial state, diet, time of administration, method of administration, drug mixture, the condition of the patient, and the extent of neurological disease.
  • the extract of the myocardium of the present invention inhibits the growth of prostate cancer cells and induces apoptosis. Therefore, the composition for treating prostate cancer according to the present invention will be very effective for the treatment of patients with prostate cancer.
  • Example 1 is a result of Alamar Blue analysis to determine the effect of the introduction of the extract of the myocardium extract obtained in Example 1 in PC-3 cells, a human prostate cancer cell line on the growth of prostate cancer cells, wherein the X-axis is the myocardium extract And the Y axis represents the survival rate of surviving human prostate cancer PC-3 cells.
  • Figure 2 is the result of Alamar Blue analysis to determine the effect of the introduction of the low myrtle skin extract obtained in Example 1 on the growth of prostate cancer cells in DU 145 cells, a human prostate cancer cell line, wherein the X-axis is the Concentration, the Y axis shows the survival rate of surviving human prostate cancer DU 145 cells.
  • Figure 3 is a flow cytometry result to determine the effect of the introduction of the extract of the myocardium epidermis obtained in Example 1 in PC-3 cells, a human prostate cancer cell line to stop the prostate cancer cell cycle and apoptosis.
  • Example 4 is a flow cytometry result to determine the effect of the introduction of the extract of the myocardium epidermis obtained in Example 1 in the human prostate cancer cell line DU 145 to stop the prostate cancer cell cycle and apoptosis.
  • 5 is a result of analysis of luciferase activity by AP-1 reporter assay method to determine the effect of the introduction of low myrtle skin extract in PC-3 human prostate cancer cell line AP-1 cell signal of prostate cancer cells .
  • the human prostate cancer cell lines PC-3 and DU 145 used in the present invention were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA) and used for experiments. Specifically, PC-3 and DU 145 cell lines were treated with DMEM (Dulbeco's Modified Eagle's Medium) medium containing 10% fetal bovine serum (FBS) (Welgene) at 37 ° C. and 5% CO 2 . It was kept as it was, and it was passaged for 2-3 days.
  • DMEM Dynabeco's Modified Eagle's Medium
  • FBS fetal bovine serum
  • PC-3 and DU 145 cells which are human prostate cancer cells, were treated with ethanol extract of the low myrtle bark for 48 hours and analyzed by Alamar Blue.
  • the Alamar Blue assay is a modified form of the MTT assay, in which a specific enzyme degrades a living cell and then measures the fluorescence intensity of the product as the compound breaks down to determine the relative number of living cells after treatment. I am an experimental method. It will be described in more detail below.
  • the plate was slowly shaken to evenly react the cells in each well, and the intensity of fluorescence was measured by Fluorescence Microplate Reader (Molecular Devices Corp.) at 590 nm while irradiating irradiated light at a wavelength of 544 nm. Survival rates of -3 and DU 145 human prostate cancer cells are also shown in FIGS. 1 and 2.
  • Flow Cytometry was performed to investigate the effect of Rhizome extract on the cell division cycle of cancer cells.
  • Flow cytometry is a method to quickly identify the characteristics of a cell as it passes through the detection zone. When staining the cell nucleus with a fluorescent material, the flow cytometry can be determined. .
  • PC-3 and DU 145 cells were seeded in 6-well plates at about 2.4 ⁇ 10 5 cells per well and incubated for 24 hours.
  • the cells were administered with 100 ⁇ g / ml of low myrtle skin extract and allowed to stand for 24 hours.
  • the culture medium in each well was removed from the 6-well plate and washed twice with PBS (phosphate-buffered saline).
  • the attached cells were suspended by treatment with trypsin-EDTA solution, transferred to microtubes and centrifuged to recover the cells.
  • the recovered cells were washed with PBS, centrifuged and suspended in cold ethanol. Cells suspended in ethanol were stored at 4 ° C to fix the cells, centrifuged and washed with PBS.
  • the washed cells were suspended in 500 ⁇ l of PI solution (50 ⁇ g / ml of Propidium Iodide, 10 ⁇ g / ml of RNase A) and stored at 37 ° C. for 30 minutes to stain nuclei, and 2 ml of PBS was added thereto. Cells suspended in solution were analyzed by flow cytometry (FACS; BD biosciences), and cell cycles and apoptosis of PC-3 and DU 145 human prostate cancer cells were shown in FIGS. 3 and 4, respectively.
  • PI solution 50 ⁇ g / ml of Propidium Iodide, 10 ⁇ g / ml of RNase A
  • the reporter assay transfects human prostate cancer cell PC-3 with each luciferase vector designed to measure the transcriptional activity of AP-1, which has a major effect on cancer cell growth.
  • the extract was treated with a low myofiformum extract at a concentration of 0 to 100 ⁇ g / ml and after 24 hours, luciferase activity was measured.
  • the degree of inhibition of AP-1 cell signal expressed in the myofibril extract of human prostate cancer cells is described in Table 3 below.
  • the cancer-treated cells treated with low myofi calli reduced the transcriptional activity of AP-1, which leads to the growth of cancer cells, compared to the control group. It was found that suppression.
  • PC-3 cells were cultured to grow 60 to 70% in a 60 mm dish, and then the low myrtle bark extract was treated at a concentration of 0 to 100 ⁇ g / ml. After 24 hours, the culture medium was removed from the cells in culture, washed with PBS, and treated with NP-40 solution to obtain cell lysate. The cell lysate was centrifuged to recover the supernatant in which the cellular protein was dissolved, and the sample was prepared for electrophoresis after measuring the concentration. The prepared samples were electrophoresed and Western blotting was performed to analyze the degree of Caspase 3 activation with Caspase 3 and Bax antibodies, and the results are shown in FIG. 6.
  • the extract of Low Myofi cava was confirmed that Caspase 3, which leads to the apoptosis of cancer cells, was activated, and it can be seen that the Myofi cava exhibits cancer cells effectively resulting in apoptosis.
  • the present inventors have confirmed that the prostate cancer treatment effect of the low myrtle bark extract through the above embodiment was prepared to prepare a prostate cancer treatment containing the low myrtle bark extract as an active ingredient as follows.
  • Vitamin D 3 0.001%
  • Vitamin D 3 0.001%

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Abstract

L'invention concerne une nouvelle utilisation d'un extrait d'ailante glanduleux, et plus particulièrement, une composition contenant un extrait de solvant organique d'ailante glanduleux pour prévenir et traiter le cancer de la prostate. La composition pour traiter le cancer de la prostate, selon l'invention, supprime efficacement la croissance des cellules du cancer de la prostate et elle induit l'apoptose, ainsi, elle peut être efficacement utilisée pour traiter et prévenir le cancer de la prostate.
PCT/KR2011/006421 2010-08-30 2011-08-30 Composition contenant un extrait de solvant organique d'ailante glanduleux pour prévenir et traiter le cancer de la prostate WO2012030150A2 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102659746A (zh) * 2012-04-19 2012-09-12 山东省医学科学院药物研究所 一种臭椿花提取物与其活性成分短叶苏木酚的制备方法及应用
KR101545507B1 (ko) * 2013-09-30 2015-08-19 서울대학교산학협력단 가중나무 추출물을 유효성분으로 하는 암질환의 치료 또는 예방용 조성물
KR20180032415A (ko) * 2016-09-22 2018-03-30 이화여자대학교 산학협력단 방사선 조사 또는 항암제에 의한 세포 손상 예방 또는 치료용 약학 조성물
CN115487179A (zh) * 2022-11-18 2022-12-20 暨南大学 抗肿瘤的联合用药物及其应用

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
H. R. PARK ET AL.: 'Aqueous root extract of Ailanthus altissima extract induced apoptosis in K562 human leukemia cells' KOREAN JOURNAL OF ORIENTAL PHYSIOLOGY & PATHOLOGY vol. 18, no. 6, 2004, pages 1728 - 1732 *
J. H. JEONG ET AL.: 'Effects on inhibition of angiogenesis in MCF-7 cells by the aqueous root extract of Ailanthus Altissima' KOREAN JOURNAL OF ORIENTAL PHYSIOLOGY & PATHOLOGY vol. 18, no. 6, 2004, pages 1613 - 1616 *
J. KIM ET AL.: 'Studies on the biological activities of the constituents of Ailanthi cortex radics III. Antitumor activities of dichloromethane fraction' KOREAN JOURNAL OF PHARMACOGNOSY vol. 28, no. 1, 1997, pages 54 - 58 *
R. J. A. VAN MOORSELAAR ET AL.: 'Angiogenesis in prostate cancer: its role in disease progression and possible therapeutic approaches' MOLECULAR AND CELLUAR ENDOCRINOLOGY vol. 197, 2002, pages 239 - 250 *
R. W. G. WATSON ET AL.: 'Targeting apoptosis in prostate cancer: focus on caspases and inhibitors of apoptosis proteins' BJU INTERNATIONAL vol. 96, no. SUPPLE, 2005, pages 30 - 34 *
S.-G. HWANG ET AL.: 'Induction of apoptosis in Jurkat T lymphocytes by extract of Ailanthus altissima' KOREAN JOURNAL OF PHARMACOGNOSY vol. 32, no. 4, 2001, pages 274 - 279 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102659746A (zh) * 2012-04-19 2012-09-12 山东省医学科学院药物研究所 一种臭椿花提取物与其活性成分短叶苏木酚的制备方法及应用
KR101545507B1 (ko) * 2013-09-30 2015-08-19 서울대학교산학협력단 가중나무 추출물을 유효성분으로 하는 암질환의 치료 또는 예방용 조성물
KR20180032415A (ko) * 2016-09-22 2018-03-30 이화여자대학교 산학협력단 방사선 조사 또는 항암제에 의한 세포 손상 예방 또는 치료용 약학 조성물
KR101975640B1 (ko) 2016-09-22 2019-05-07 이화여자대학교 산학협력단 방사선 조사 또는 항암제에 의한 세포 손상 예방 또는 치료용 약학 조성물
CN115487179A (zh) * 2022-11-18 2022-12-20 暨南大学 抗肿瘤的联合用药物及其应用

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