WO2012030142A2 - Composition pour traiter le cancer de la prostate comprenant un extrait de nard - Google Patents

Composition pour traiter le cancer de la prostate comprenant un extrait de nard Download PDF

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Publication number
WO2012030142A2
WO2012030142A2 PCT/KR2011/006409 KR2011006409W WO2012030142A2 WO 2012030142 A2 WO2012030142 A2 WO 2012030142A2 KR 2011006409 W KR2011006409 W KR 2011006409W WO 2012030142 A2 WO2012030142 A2 WO 2012030142A2
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WO
WIPO (PCT)
Prior art keywords
prostate cancer
extract
composition
cells
persimmon
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PCT/KR2011/006409
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English (en)
Korean (ko)
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WO2012030142A3 (fr
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황성연
정경채
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주식회사 한국전통의학연구소
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Publication of WO2012030142A2 publication Critical patent/WO2012030142A2/fr
Publication of WO2012030142A3 publication Critical patent/WO2012030142A3/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)
    • A61K36/84—Valerianaceae (Valerian family), e.g. valerian
    • 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 persimmon extract.
  • the present invention relates to a therapeutic composition containing as an active ingredient extracts persimmon extract showing 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).
  • a sense songhyang (Nardostachytis Rhizoma) is used as the root of a perennial plant of a sense songhyang (nardostachys Batalin chinensis) stomach pain, stomach cramps, thoracoabdominal coupon, neurogenic gastrointestinal disorders, vomiting, headache, or the like, respectively.
  • Nardostachys jatamansi (NJ) has been used extensively as a tonic, irritant and anticonvulsant in several Asian countries, as well as to treat epilepsy, pathological excitement, palpitations and spasms (Bagchi, A., et al. , Planta Med., 57, 9697 (1991)).
  • Perennial perennial persimmon roots include 1-aristol-2-one, nardostachone, 1,8,9,10-tetrad-hydroaristolan-2- components such as one) are contained.
  • the self-acting effect of the sensational sensation relieves pain caused by abdominal pain and chest abdominal pain, and is also applied to neuropathic pain and headache. Wash each affected area by adding water to it. It acts on the central nervous system and has a calming effect, such as heel chogeun ( ⁇ ⁇ ).
  • heart rate control is remarkable, and smooth muscle spasm of the bronchus, small intestine, large intestine, and uterus is released. Minor but also antibacterial.
  • the present inventors completed the present invention by confirming that the extract of the extract of Persimmon can effectively kill the prostate cancer cells while studying the herbal medicine for the extract.
  • An object of the present invention to provide a composition for the treatment of prostate cancer containing the extract persimmon flavor as an active ingredient.
  • the present invention provides a composition for the prevention and treatment of prostate cancer containing an organic solvent extract of Nardostachyos Rhizoma 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, Most preferably, the ethanol is 95%.
  • the composition of the present invention contains the extract of Persimmon as an active ingredient, and may further include a pharmaceutically acceptable carrier or diluent.
  • the persimmon extract of the present invention is possible at any site of persimmon, but is preferably extracted from the root.
  • 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.
  • ethanol 1 to 5 times, preferably 3 times, 95% ethanol is added to the dried persimmon fragrance or powder, and 10 to 100 hours, preferably 15 at a temperature of 20 to 100 ° C, preferably 40 to 60 ° C.
  • 10 to 100 hours preferably 15 at a temperature of 20 to 100 ° C, preferably 40 to 60 ° C.
  • To 40 hours, more preferably, for 24 hours after extraction can be filtered to prepare an ethanol extract of persimmon flavor.
  • 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 contains a persimmon fragrance 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 extracts of the extracts kill 86% and 82% of PC-3 or DU 145 prostate cancer cells at 100 ⁇ g / ml, respectively.
  • the above results demonstrate that the extract of Persimmon 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 sex, age, affection status, diet, time of administration, method of administration, drug mixture, the condition of the patient and the incidence of neurological disease.
  • the extract of saenghyanghyang can prevent prostate cancer cells of PC-3 and DU 145 cells, which are human prostate cancer cell lines, in a specific cell cycle to stop cell division in apoptosis (apoptosis process). apoptosis) process (see FIGS . 3 and 4 ).
  • the extract of Persimmon extract activates Caspase 3, which affects NF- ⁇ B cell signal of prostate cancer cells and induces apoptosis (see FIG. 5 ).
  • the extract of Seng Songhyang has a prostate cancer treatment effect.
  • the extract 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 from Example 1 in PC-3 cells, human prostate cancer cell line on the growth of prostate cancer cells, wherein the X-axis is the concentration of the 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 extract of the extract from 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 of the extract , Y axis shows 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 sensation incense obtained in Example 1 in PC-3 cells, a human prostate cancer cell line on the prostate cancer cells to stop the cell cycle and apoptosis.
  • Figure 4 is a flow cytometry result to determine the effect of the introduction of the extract of the extract from Example 1 in the human prostate cancer cell line DU 145 to stop the prostate cancer cells cell cycle and apoptosis.
  • FIG. 5 shows the activation of Caspase 3 to induce quantitative changes and apoptosis of I- ⁇ B ⁇ protein in order to investigate the effect of the introduction of the extract of Persimmon on the NF- ⁇ B signal of prostate cancer cells in PC-3, a human prostate cancer cell line.
  • 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
  • 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 effects of the extract of Persimmon 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.
  • Cells were administered with 100 ⁇ g / ml of Persimmon 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).
  • 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 cancer cells treated with the extracts of saccharin extract were found to be stationary in a specific cell cycle compared to the control group, and the apoptosis process, which is an apoptosis process, was in progress. From this, it can be seen that the persimmon extract stops the division of cancer cells and leads to apoptosis.
  • PC-3 cells were cultured to grow 60-70% in a 60 mm dish, and then the persimmon extract was treated with a concentration of 0-100 ⁇ 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 amount of I- ⁇ B ⁇ protein with I- ⁇ B ⁇ antibody, and the results are shown in FIG. 5.
  • the extract of S. eryngii inhibits prostate cancer cell division by increasing the amount of I- ⁇ B ⁇ protein and inhibiting NF- ⁇ B activity.
  • PC-3 cells were cultured to grow 60-70% in a 60 mm dish, and then the persimmon extract was treated with a concentration of 0-100 ⁇ 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 the Caspase 3 antibody, and the results are shown in FIG. 5.
  • the extract of S. eryngma was confirmed that Caspase 3, which leads to apoptosis of cancer cells, was activated, and it can be seen that the sensational aroma effectively leads to apoptosis of cancer cells, thereby exhibiting an anticancer effect.
  • the present inventors have confirmed that the prostate cancer treatment efficacy of the extract persimmon through the embodiment of the prostate cancer to prepare a prostate cancer treatment containing the extract as an active ingredient as follows.
  • Vitamin D 3 0.001%
  • Vitamin D 3 0.001%

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Abstract

Cette invention concerne une nouvelle utilisation de l'extrait de nard, et plus particulièrement, une composition pour prévenir et traiter le cancer de la prostate contenant un extrait de nard obtenu par un procédé au solvant organique. La composition pour traiter le cancer de la prostate selon la présente invention supprime efficacement la croissance des cellules cancéreuses prostatiques et induit l'apoptose, et peut, par conséquent, être utilisée efficacement pour traiter et prévenir le cancer de la prostate.
PCT/KR2011/006409 2010-08-30 2011-08-30 Composition pour traiter le cancer de la prostate comprenant un extrait de nard WO2012030142A2 (fr)

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WO2012030142A3 WO2012030142A3 (fr) 2012-05-31

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104666411A (zh) * 2015-02-06 2015-06-03 浙江大远智慧制药工程技术有限公司 一种丹红注射液的醇沉工艺改进方法
CN105126059A (zh) * 2015-10-03 2015-12-09 南京多宝生物科技有限公司 一种含生姜的植物组合物提取方法和应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004112692A2 (fr) * 2003-05-19 2004-12-29 Amazon Biotech Inc. Compositions therapeutiques a base de plantes medicinales

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004112692A2 (fr) * 2003-05-19 2004-12-29 Amazon Biotech Inc. Compositions therapeutiques a base de plantes medicinales

Non-Patent Citations (3)

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Title
K. TAKEYA ET AL.: 'Isolation and structural determination of antitumor substances from natural products using bio-active screening tests' JOURNAL OF ORIENTAL BOTANIC RESEARCH vol. 6, no. 1, 1993, pages 45 - 51 *
S. BAEK ET AL.: 'Antioxidant and anti-inflammatory effect of Nardostachys chinensis in IFN-gamma/LPS-stimulated pertioneal macrophage' KOREAN JOURNAL OF ORIENTAL PHYSIOLOGY AND PATHOLOGY vol. 23, no. 4, 2009, pages 853 - 859 *
S. H. YOON ET AL.: 'Extracelluar signal-regulated kinase (ERK) is required for water extract of Nardostachys chinensis-induced differentiation in HL-60 cells' KOREAN JOURNAL OF ORIENTAL PHYSIOLOGY AND PATHOLOGY vol. 20, no. 5, 2006, pages 1315 - 1320 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104666411A (zh) * 2015-02-06 2015-06-03 浙江大远智慧制药工程技术有限公司 一种丹红注射液的醇沉工艺改进方法
CN105126059A (zh) * 2015-10-03 2015-12-09 南京多宝生物科技有限公司 一种含生姜的植物组合物提取方法和应用

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