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WO2017119767A1 - Composition pour inhiber la résistance aux médicaments anticancéreux contenant un ginsénoside rare csh1 (rg6) en tant que substance active. - Google Patents

Composition pour inhiber la résistance aux médicaments anticancéreux contenant un ginsénoside rare csh1 (rg6) en tant que substance active. Download PDF

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WO2017119767A1
WO2017119767A1 PCT/KR2017/000198 KR2017000198W WO2017119767A1 WO 2017119767 A1 WO2017119767 A1 WO 2017119767A1 KR 2017000198 W KR2017000198 W KR 2017000198W WO 2017119767 A1 WO2017119767 A1 WO 2017119767A1
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csh1
taxol
drug resistance
cells
cancer
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Korean (ko)
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박범준
송규용
조수현
이지현
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University Industry Cooperation Foundation of Pusan National University
Industry and Academy Cooperation In Chungnam National University
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University Industry Cooperation Foundation of Pusan National University
Industry and Academy Cooperation In Chungnam National University
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Priority claimed from KR1020160002619A external-priority patent/KR101732876B1/ko
Application filed by University Industry Cooperation Foundation of Pusan National University, Industry and Academy Cooperation In Chungnam National University filed Critical University Industry Cooperation Foundation of Pusan National University
Publication of WO2017119767A1 publication Critical patent/WO2017119767A1/fr
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
    • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate

Definitions

  • the present invention relates to a composition for inhibiting anticancer drug resistance, containing the rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • Renal cell carcinoma is a human malignant tumor of the kidney near the adrenal gland.
  • kidney cancer is known to develop for a variety of reasons, such as aging and smoking, stress is also a major cause. Serum cortisol levels are increased in kidney cancer patients. In this regard, the effect of stress hormone suppression on the immune system is generally known, but this is not enough to account for stress-induced cancer.
  • Several epidemiological analyzes have shown that stress can increase the incidence of several types of human cancer, including colorectal and lung cancer.
  • Panax Ginseng has long been used to treat many types of human diseases, including cancer and kidney dysfunction. Recently, isolated ginsenoisdes have been reported to be effective in the immune system. However, the mechanism of molecular biological action of ginsenosides on human cancers has not been identified to date. Therefore, it is very meaningful to elucidate the mechanism and effect of ginsenosides on human cancer.
  • the present invention provides a pharmaceutical composition for inhibiting anticancer drug resistance or a pharmaceutical composition for preventing or treating cancer, containing the rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • the present invention is to provide a food composition for inhibiting anticancer drug resistance containing rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • the present invention provides a pharmaceutical composition for inhibiting cancer drug resistance containing rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • the present invention also provides a pharmaceutical composition for preventing or treating cancer containing the rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • the present invention also provides a food composition for inhibiting anticancer drug resistance, containing the rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • the present invention relates to a food composition for inhibiting anticancer drug resistance containing rare ginsenoside CSH1 (Rg6) as an active ingredient, wherein CSH1 (Rg6) promotes the binding of BRCA1 and Rad51 and expresses stress hormone-induced ⁇ -tubulin expression. It was confirmed that the effect of inhibiting the resistance to Taxol in cancer cells. Therefore, the CSH1 (Rg6) is expected to be usefully used in drugs, anticancer drugs or foods that inhibit anticancer drug resistance in anticancer treatment.
  • CSH1 (Rg6) rare ginsenoside CSH1 (Rg6) as an active ingredient, wherein CSH1 (Rg6) promotes the binding of BRCA1 and Rad51 and expresses stress hormone-induced ⁇ -tubulin expression. It was confirmed that the effect of inhibiting the resistance to Taxol in cancer cells. Therefore, the CSH1 (Rg6) is expected to be usefully used in drugs, anticancer drugs or foods that inhibit anticancer drug resistance in anticancer treatment.
  • GR 2 shows that stress hormones increase MTOC through the Glucocorticoid Receptor (GR).
  • Cortisol clearly induced ⁇ -tubulin in several cell lines. Each cell was treated for 72 hours with the indicated dose of cortisol. Western blots were performed to measure ⁇ -tubulin expression. Actin was used as a loading control.
  • GHs induce ⁇ -tubulin independently of FST. A498 (VHL deficient cell line) was incubated with cortisone (5 ⁇ M) and cortisol (5 ⁇ M) for 24 hours with or without FST (3 ⁇ M). BRCA1 expression was increased by FST but did not affect the reduction of BRCA1 expression by GH.
  • p53 expression was not altered by FST or GH. Actin was used as a loading control.
  • the number of MTOCs was increased by cortisone treatment (5 ⁇ M) in VHL-intact C2V cells. About 50 cells were counted for each condition to determine the average MTOC number and plotted. Cells were stained with anti- ⁇ -tubulin-antibody (red) to detect MTOC and DAPI (blue) to detect DNA.
  • GR and cortisone can induce MTOC amplification. After GR transfection or cortisone (5 ⁇ M) treatment, HEK293 cells were stained with anti- ⁇ -tubulin-antibody (red) and DAPI (blue).
  • (f) shows the additional effect of GH and GR on MTOC amplification. About 50 cells were counted for each condition to determine the MTOC number.
  • (g) In VHL-positive C2V cells, the overexpressed GR showed Taxol resistance. GR was transfected into C2V cells and treated with Taxol (3 ⁇ M). After 72 hours, cell viability was measured by MTT assay.
  • (h) GR blocks Rad51-mediated Taxol-sensitivity. In VHL-deficient A498, re-sensitivity to Taxol via Rad51 overexpression was lost by GR overexpression. A498 cells were transfected with GR and / or Rad51 and incubated with Taxol for 72 hours. Cell viability was measured by MTT assay.
  • GR inhibition blocks MTOC amplification (a and b) GR antagonist Progesterone (PGT) blocked MTOC in ACHN cells. PGT (5 ⁇ ) treated cells blocked MTOC amplification. About 80 cells were counted for each condition to confirm the mean MTOC number (b) and photographed (a). Cells were stained with anti- ⁇ -tubulin-antibody (red) to detect MTOC and DAPI (blue) to detect DNA. (c) PGT treatment inhibited stress hormone-induced ⁇ -tubulin induction. Western blot analysis was performed with the indicated antibodies. Actin was used as a loading control. (d) Ketoconazole (KCZ), another chemical antagonist of GR, also blocked stress hormone effects in ACHN cells.
  • PGT Progesterone
  • FIG. 5 shows the chemical structures of the rare ginsenosides CSH1 (RG6) and RG3, estrogen and cortisone.
  • Figure 6 shows the effect of rare ginsenosides on Taxol-induced cell death.
  • (a) shows the effect of several ginsenosides on Taxol-induced cell death. In C2 cells, only CSH1 (Rg6) shows Taxol sensitive effect. Ginsenosides (5 ⁇ M) and Taxol (3 ⁇ M) were treated respectively. After 72 hours, cell viability was measured by MTT assay.
  • PGT (5 ⁇ M), CSH1 (5 ⁇ M), cortisol (5 ⁇ M) and cortisone (5 ⁇ M) were treated for 72 hours.
  • CSH1 could block GR or cortisone induced MTOC amplification. About 50 cells were counted to confirm the mean MTOC number and photographed. GFP-labeled GR was transfected into HEK 293 cells. CSH1 (5 ⁇ M) and cortisone (5 ⁇ M) were treated. Cells were stained with anti- ⁇ -tubulin-antibody (red) and DAPI (blue).
  • Figure 7 shows the chemical structures of CSH1-related ginsenosides CSH2 to CSH4.
  • FIG. 8 shows that two normal human fibroblasts were cultured for one month with or without the addition of cortisol and CSH1 at low serum concentrations to confirm that continuous treatment of stress hormones could induce transformation. to be.
  • ERE-luciferase activity was measured by the ERE-luciferase vector and ER- ⁇ .
  • Similar results were obtained with MCF-7 cells.
  • CSH1 effectively inhibited ERE luciferase activity.
  • ERE-luciferase vectors were transfected into ER ⁇ -positive MCF7 cells and ER ⁇ -negative MDA-MB-468 cells. ERE-luciferase analysis was performed 4 hours after each compound treatment.
  • e In VHL-negative C2 cells, Taxol-induced cell death by CSH1 showed a concentration dependent sensitivity similar to FST.
  • FST FST
  • CSH1 5 ⁇ M
  • CSH3 5 ⁇ M
  • IF staining was performed.
  • Cells were stained with anti- ⁇ -tubulin-antibody (red) and DAPI (blue).
  • FIG. 10 is a schematic diagram showing that the GR pathway is activated in response to stress hormones, and MTOC amplification and chromosomal instability are induced under stress conditions. It is one of the estimated tumorigenic mechanisms for stress-induced cancer. However, unlike the ER- ⁇ pathway, GR is not regulated by VHL. CSH1, a rare ginsenoside, has a steroid-like backbone, which blocks GR as well as ER- ⁇ pathway-induced MTOC amplification and Taxol sensitivity. Therefore, it can be used as a cancer prevention strategy and anticancer drug.
  • the present inventors therefore focused on the tumor forming effects of stress hormones, in particular on MTOC amplification and anticancer drug resistance. Since several ginsenosides have stress hormone-related chemical structures, the effects of ginsenosides on MTOC amplification and Taxol resistance have been investigated and the present invention has been completed.
  • the present invention provides a pharmaceutical composition for inhibiting anticancer drug resistance, containing the rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • the anticancer drug resistance may be induced by a stress hormone, but is not limited thereto.
  • the rare ginsenoside CSH1 (Rg6) inhibits ⁇ -tubulin expression induced by stress hormones, promotes binding of BRCA1 and Rad51, and amplifies the microtubule organizing center (MTOC). Can be suppressed.
  • the stress hormone may be Cortisone or Cortisol, but is not limited thereto.
  • the anticancer agent may be Taxol, but is not limited thereto.
  • the present invention also provides a pharmaceutical composition for preventing or treating cancer containing the rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • the cancer may be cancer having anticancer drug resistance, but is not limited thereto.
  • the anticancer agent may be Taxol, but is not limited thereto.
  • the cancer may be kidney cancer, lung cancer, colon cancer or breast cancer, but is not limited thereto.
  • the pharmaceutical composition is any one selected from the group consisting of injections, granules, powders, tablets, pills, capsules, suppositories, gels, suspensions, emulsions, drops or solutions according to conventional methods
  • the pharmaceutical composition is a suitable carrier, excipient, disintegrant, sweetener, coating agent, swelling agent, lubricants, lubricants, flavoring agents, antioxidants, buffers, bacteriostatic agents commonly used in the manufacture of pharmaceutical compositions It may further comprise one or more additives selected from the group consisting of diluents, dispersants, surfactants, binders and lubricants.
  • the carriers, excipients and diluents are lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil can be used, and solid preparations for oral administration include tablets, pills, powders, granules, capsules.
  • solid preparations may be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like in the composition.
  • excipients such as starch, calcium carbonate, sucrose or lactose, gelatin and the like
  • lubricants such as magnesium styrate and talc may also be used.
  • Oral liquid preparations include suspensions, solvents, emulsions, syrups, and the like, and may include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin.
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like.
  • non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.
  • Witsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like may be used as the base material of the suppository.
  • the pharmaceutical composition is intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, nasal, inhaled, topical, rectal, oral, intraocular or intradermal Via the route can be administered to the subject in a conventional manner.
  • the preferred dosage of the pharmaceutical composition may vary depending on the condition and weight of the subject, the type and severity of the disease, the drug form, the route of administration, and the duration, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention, but not limited thereto, the daily dosage may be 0.01 to 200 mg / kg, specifically 0.1 to 200 mg / kg, more specifically 0.1 to 100 mg / kg. Administration may be administered once a day or divided into several times, thereby not limiting the scope of the invention.
  • the 'subject' may be a mammal including a human, but is not limited thereto.
  • the present invention provides a food composition for inhibiting anticancer drug resistance containing rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • Rg6 rare ginsenoside CSH1
  • the anticancer drug resistance may be induced by a stress hormone, but is not limited thereto.
  • the rare ginsenoside CSH1 (Rg6) inhibits ⁇ -tubulin expression induced by stress hormones, promotes binding of BRCA1 and Rad51, and amplifies the microtubule organizing center (MTOC). Can be suppressed.
  • the stress hormone may be Cortisone or Cortisol, but is not limited thereto.
  • the anticancer agent may be Taxol, but is not limited thereto.
  • the present invention also provides a food composition for preventing or improving cancer containing rare ginsenoside CSH1 (Rg6) as an active ingredient.
  • Rg6 rare ginsenoside CSH1
  • the cancer may be cancer having anticancer drug resistance, but is not limited thereto.
  • the anticancer agent may be Taxol, but is not limited thereto.
  • the cancer may be kidney cancer, lung cancer, colon cancer or breast cancer, but is not limited thereto.
  • the food composition of the present invention is variously used as a food composition for inhibiting anticancer drug resistance, the food composition comprising the composition of the present invention as an active ingredient, various foods, for example, beverages, gum, tea, vitamin complex, It may be prepared in the form of powder, granules, tablets, capsules, sweets, rice cakes, bread and the like. Since the food composition of the present invention has little toxicity and side effects, it can be used with confidence even for long-term use for the purpose of prevention. When the composition of the present invention is included in a food composition, the amount may be added at a ratio of 0.1 to 100% of the total weight.
  • natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, sucrose and the like, and common sugars such as polysaccharides, dextrins and cyclodextrins, and sugar alcohols such as xylitol, sorbitol, and erythritol. can do.
  • flavourant examples include natural flavourant (tautin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).
  • the food composition is a variety of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH regulators Stabilizing agents, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, etc. These components may be used independently or in combination
  • the ratio of such additives is not so critical, but 100 weight of the composition of the present invention. It is generally selected in the range of 0.1 to 100 parts by weight per part.
  • ACHN (VHL +) and A498 (VHL ⁇ ) cells were purchased from the Korean Cell Line Bank.
  • A549, HEK293, MCF-7 and MDA-MB-468 were purchased from the American Cell Line Bank (ATCC, Manassas, VA).
  • Other renal cancer cell lines (UMRC2; C2, UMRC2 / VHL; C2V) are described by Dr. Jung, YJ (Pusan University).
  • Human fibroblasts were purchased from Coriell Cell Repositories (New Jersey, USA).
  • ACHN, A498, HEK293, MCF-7 and MDA-MB-468 were maintained in liquid DMEM medium containing 10% FBS and 1% antibiotic in a 37 ° C. growth chamber.
  • HCT116 cell line Obtained from Vogelstein B (Johns Hopkins University). A549, HCT116 was maintained in RPMI-1640 with 10% FBS and antibiotics. Estrogen (250155), Fulvestrant (I4409), Taxol (T7402), Ketoconazole (UC280), Progesterone (P0130), Cortisone (C2755) and Cortisol; H4001 ) was purchased from Sigma (Missouri, USA). Antibodies against Actin (sc-1616), ER- ⁇ (sc-8002), GFP (sc-7392) and HA (sc-9996) were purchased from Santa Cruz (California, USA).
  • Anti- ⁇ -tubulin (T6557) was purchased from Sigma (Missouri, USA) and anti-glucocorticoid receptor (GR) Ab (12041) was purchased from Cell signaling (Massachusetts, USA). Rad51 (05-530), BRCA1 (07-434) were purchased from Milliopore (Darmstadt, Germany).
  • GFP-tagged GR (GR-GFP) and HA-tagged Rad51 (Rad51-HA) vectors were purchased from Addgene.
  • pVHL animal cell expression vector is Obtained from Jung, YJ (Pusan University).
  • Transfection for animal cell expression of the vectors was performed using a Jetpei transfection agent (Polyplus New York, USA). Briefly, the vector (1.5 ⁇ g) was mixed with 1.5 ⁇ l Jetpei reagent dissolved in 150 mM NaCl solution. The mixture was reacted at room temperature for 15 minutes. After the reaction, the mixture was added to the cells. After 3 hours, serum-free medium was replaced with medium containing 10% FBS.
  • Proteins were extracted from cells using RIPA buffer (50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.1% SDS and 10% sodium deoxycholate). Samples were applied to SDS-PAGE and transferred to PVDF membrane. The blotted membrane was reacted with the primary antibody for 1 hour at 4 ° C., and HRP-conjugate species-matched secondary antibodies were reacted for 1 hour at room temperature. Peroxidase activity was detected through chemi-luminescence with an ECL kit (Intron, Seoul, Korea).
  • IP immunoprecipitation
  • Cells were placed in a cover glass and transfected with the indicated vectors or treated with the indicated compounds. After fixing for 30 minutes with Me-OH, cells were reacted with blocking buffer [PBS + anti-human-Ab (1: 500)] for 1 hour. After washing with PBS, cells were reacted for 4 hours with Anti- ⁇ -tubulin antibody (1: 100 to 200) dissolved in blocking buffer, and FITC-conjugated or Rhodamine-conjugated secondary antibody (1) dissolved in blocking buffer. : 500) for 2 hours. Nuclei were stained with DAPI. After rinsing with PBS, the cover glass was mounted in mounting solution (Vector Laboratories, Cambridgeshire, UK). Immunofluorescence signals were detected by fluorescence microscopy (Zeiss, Jena, Germany).
  • MTT assay To measure cell viability, cells were treated with the indicated compounds for 4 days.
  • Luciferase Luciferase
  • ERE-Luc vectors were transfected into cells for 24 hours and cells were treated with the indicated compounds. After washing with wash buffer (Promega, Wisconsin, USA), cells were lysed with lysis buffer (Promega, Wisconsin, USA). Luciferase activity was measured by a luminometer (MicroDigital, Gyeonggi-do, South Korea).
  • Cortisol a stress hormone and related hormones (cortisone and aldosterone), originated from cholesterol and exhibited a chemical structure similar to estrogen, so their biological effects on Taxol-induced cell death were tested. Although not aldosterone, cortisone and cortisol showed Taxol resistance in both RCC cell lines similar to estrogens (FIG. 1A). Their inhibitory effect was dose-dependent (FIG. 1B). Stress hormones can affect many types of tissues and cells. We have identified the effects of glucocorticoid hormones in lung and colorectal cancer cell lines, and cortisol and cortisone are Taxol-induced cells in a dose-dependent manner. Results similar to inhibiting death were obtained (FIG. 1C).
  • Cortisol and cortisone are glucocorticoid hormones, and their signaling is mediated by glucocorticoid receptors (GR), so we have identified the association of GR to MTOC amplification. Transfection with GR alone was able to increase MTOC numbers as well as cortisone treatment (FIGS. 2E and 2F). GR can also block Taxol-induced cell death (FIG. 2G). Thus, we identified the effect of GR on Rad51-mediated Taxol sensitivity. Previous studies have found that Rad51 overexpression can regain sensitivity in Taxol-resistant ER- ⁇ elevated cells and VHL deficient cell lines.
  • GR overexpression could reduce Rad51 to block Rad51-mediated Taxol re-sensitivity (FIGS. 2H and 2I).
  • GR overexpression and cortisone treatment reduced endogenous Rad51 expression (FIG. 2J).
  • GR expression did not change with VHL status.
  • VHL overexpression or knock down did not affect GR expression, and FST did not affect GR expression (FIG. 2K).
  • stress hormones induce MTOC amplification, and ⁇ -tubulin elevation is caused by GR through VHL independent mechanism.
  • PGT progesterone
  • KCZ ketoconazole
  • CSH1 showed a similar effect in A498 cells.
  • CSH2 to CSH4 FIG. 7
  • CSH1-related ginsenosides CSH2 to CSH4; FIG. 7
  • CSH1 and CSH3 have increased Taxol sensitivity similar to FST.
  • C2V taxol sensitive cell line
  • the present inventors confirmed the effect of CSH1 on ⁇ -tubulin induction by cortisol and cortisone.
  • CSH1 inhibited stress hormone-induced ⁇ -tubulin expression as well as MTOC amplification in human lung cancer cell line A549 (FIGS. 6B and 6C).
  • the present inventors confirmed the beneficial effect of CSH1 in human colon cancer cell line HCT116. The results indicate that CSH1 can inhibit MTOC amplification regardless of cancer cell type.
  • CSH1 inhibited GR expression and inhibited the reduction of Rad51 by GR-overexpression (FIG. 6D).
  • CSH1 promoted the interaction of BRCA1 and Rad51 and was able to overcome their binding broken by GR (FIG. 6E).
  • the inventors confirmed the number of MTOCs. As expected, the increase in MTOC by cortisone or GR overexpression was completely inhibited by CSH1 treatment (FIGS. 6F and 6G). Since CSH1 blocks MTOC amplification and inhibition of ⁇ -tubulin regulation, it is expected that the compound may be usefully used for cancer prevention.
  • Example 8> shows a similar effect to FST CSH1
  • CSH1 exhibits sensitivity to Taxol in C2 cells, thus confirming the possibility that CSH1 could replace FST.
  • estrogen-induced cell proliferation was observed. Similar to FST in ER- ⁇ positive MCF-7 cell line, CSH1 was able to block estrogen-induced cell proliferation (FIG. 9A). In addition, CSH1 was able to inhibit estrogen-induced- ⁇ -tubulin and ER- ⁇ (FIG. 9B). However, they did not alter basic Rad51 expression (FIG. 9B). In addition, ER- ⁇ -mediated transcriptional activity was reduced by CSH1 in exogenous ER- ⁇ transfected 293 cells (FIG. 9C).

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Abstract

La présente invention concerne une composition pharmaceutique pour inhiber la résistance aux médicaments anticancéreux qui contient un ginsénoside rare CSH1 (RG6) en tant que substance active. Il a été observé que CSH1 (Rg6) stimule la liaison de BRCA1 et Rad51, inhibe l'expression de γ-tubuline induite par l'hormone de stress, et inhibe la résistance au taxol dans des cellules cancéreuses. Par conséquent, CSH1 (Rg6) devrait être utile pour des médicaments pour inhiber la résistance à des médicaments anticancéreux en thérapie anticancéreuse, ou des médicaments ou des aliments anticancéreux.
PCT/KR2017/000198 2016-01-08 2017-01-06 Composition pour inhiber la résistance aux médicaments anticancéreux contenant un ginsénoside rare csh1 (rg6) en tant que substance active. Ceased WO2017119767A1 (fr)

Applications Claiming Priority (4)

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KR10-2016-0002619 2016-01-08
KR1020160002619A KR101732876B1 (ko) 2016-01-08 2016-01-08 희귀 진세노사이드 CSH1(Rg6)을 유효성분으로 함유하는 항암제 내성 억제용 약학조성물
KR20170001813 2017-01-05
KR10-2017-0001813 2017-01-05

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US20150297727A1 (en) * 2013-04-28 2015-10-22 Fujian South Pharmaceutical Co. Ltd. Saponin nano micelle, preparing method, application and pharmaceutical composition thereof
CN103230406A (zh) * 2013-05-08 2013-08-07 江苏省中医药研究院 人参皂苷Rg6在制备治疗淋巴瘤药物中的应用

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KR101732876B1 (ko) 희귀 진세노사이드 CSH1(Rg6)을 유효성분으로 함유하는 항암제 내성 억제용 약학조성물

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