WO2015102448A1 - Promédicament hybride anticancéreux produisant simultanément le cinnamaldéhyde et le méthide de quinone et son procédé de préparation - Google Patents

Promédicament hybride anticancéreux produisant simultanément le cinnamaldéhyde et le méthide de quinone et son procédé de préparation Download PDF

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WO2015102448A1
WO2015102448A1 PCT/KR2015/000067 KR2015000067W WO2015102448A1 WO 2015102448 A1 WO2015102448 A1 WO 2015102448A1 KR 2015000067 W KR2015000067 W KR 2015000067W WO 2015102448 A1 WO2015102448 A1 WO 2015102448A1
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cancer
formula
compound
cinnamic aldehyde
anticancer prodrug
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Korean (ko)
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권병목
이동원
노정연
권병수
한동초
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Korea Research Institute of Bioscience and Biotechnology KRIBB
Industry Academic Cooperation Foundation of Chonbuk National University
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Korea Research Institute of Bioscience and Biotechnology KRIBB
Industry Academic Cooperation Foundation of Chonbuk National University
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Priority to US15/109,621 priority Critical patent/US9950071B2/en
Priority claimed from KR1020150000660A external-priority patent/KR101688887B1/ko
Publication of WO2015102448A1 publication Critical patent/WO2015102448A1/fr
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    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
    • C07F5/02—Boron compounds
    • C07F5/025—Boronic and borinic acid compounds

Definitions

  • the present invention relates to a hybrid anticancer prodrug which simultaneously produces cinnamic aldehyde and quinone metide, and to a hybrid anticancer prodrug which induces death of cancer cells while increasing oxidative stress by complementary synergistic action, and a preparation method thereof.
  • Cancer is defined as “a tumor formed by undifferentiated cells that proliferate and ignore indefinitely and ignore the necessary conditions in tissues, unlike normal cells that can proliferate and inhibit regularly according to the needs of the individual”.
  • the cell has been transformed into cancer cells by a change in the genes in the cell for some reason. Cancer accounts for 13% of all deaths worldwide and can occur regardless of gender or age, and it is a serious disease that is the second leading cause of death among the world's diseases. Research is ongoing. In these studies, the development of effective anticancer drugs with less side effects and resistance to the disease is required due to the variety of cancers and the diversification of pathogenesis, and new anticancer drugs are continuously being released.
  • the stimulus-sensitization system responds to external environment such as pH, temperature, ionic strength, electric field, magnetic field, light, ultrasound, etc.
  • external environment such as pH, temperature, ionic strength, electric field, magnetic field, light, ultrasound, etc.
  • a stimulus response system is mainly used in a release control system that not only protects the drug but also regulates the release rate of the drug or induces the drug to stay at a specific site.
  • the pH of the cancer site is very widely used because there is a variety of functional groups sensitive to the pH different from the general body pH (7.4 ⁇ 0.04).
  • Cinnam aldehyde is the main active ingredient of cinnamomum cassia, a Lauraceae plant, which has been used to treat indigestion, gastritis, blood circulation disorder and inflammation in both the East and West. ) Is the main component. Cinnamic aldehydes contain ⁇ , ⁇ -carbonyl, known as the Micheal receptor pharmacophore, and produce reactive oxygen species (ROS) that reduce the mitochondrial membrane potential. Anti-cancer ability has been demonstrated through a caspase-dependent mechanism as a substance that induces apoptosis through the release of cytochrome C from cells to cytosol.
  • ROS reactive oxygen species
  • quinone methide reacts with GSH (glutathione, glutathione), an essential antioxidant enzyme in cancer cells, and is known to cause anticancer effects by lowering antioxidant levels and increasing oxidative stress.
  • GSH glutathione, glutathione
  • the present inventors have studied to prepare a hybrid anticancer prodrug that synergizes to enhance short-term retention time and efficacy of cinnamic aldehyde which induces apoptosis, and to enhance its effect.
  • QCA (4- (1,3,2-dioxaborinan-2-yl) benzyl ((5-methyl-2-styryl-1,3-dioxan-5-yl) methyl), a prodrug that simultaneously produces quinone metide carbonate) and 4- (1,3,2-dioxaborinan-2-yl) benzyl ((5-methyl-2- (2-benzoyloxyphenyl) vinyl-1,3-dioxan-5-yl) methyl) carbonate (QBCA)
  • the QCA and QBCA prepared above enhance the retention time of cinnamic aldehyde, and furthermore, selectively act on cancer cells to maximize the anticancer effect while minimizing side effects, confirming the potential as a new anticancer drug, and the present invention. was completed.
  • the present invention is a hybrid anticancer prodrug which simultaneously generates cinnamic aldehyde and quinone metide, and aims to provide a hybrid anticancer prodrug which induces apoptosis of cancer cells while increasing oxidative stress by complementary synergy. .
  • an object of the present invention is to provide a composition for the prevention or treatment of cancer comprising the hybrid anti-cancer prodrug as an active ingredient.
  • the present invention provides a hybrid anticancer prodrug which simultaneously produces cinnamaldehyde and quinone metide, and a hybrid anticancer prodrug which induces apoptosis of cancer cells while increasing oxidative stress by complementary synergy.
  • the present invention also provides a pharmaceutical composition for the prevention or treatment of cancer comprising the hybrid anticancer prodrug as an active ingredient.
  • the present invention also provides a food composition for the prevention or improvement of cancer comprising the hybrid anticancer prodrug as an active ingredient.
  • the hybrid anticancer prodrug of the present invention is to release quinone methide and cinnamic aldehyde sequentially by H 2 O 2 and acidic pH to alkylate the antioxidant GSH by release of quinone methyst to inhibit the antioxidant system and oxidative stress ROS is produced and accumulated by the release of cinnamic aldehyde, which promotes apoptosis, resulting in a synergistic anticancer effect through a dual stimulation response and sequential treatment in a cancer cell-specific manner, which is useful as an anticancer agent.
  • Example 1 is a diagram showing a 1 H-NMR spectrum of (4- (1,3,2-dioxaborinan-2-yl) phenyl) methanol (1) prepared in Example 1-1 of the present invention.
  • Figure 2 is a diagram showing the 1 H-NMR spectrum of the cinnamic aldehyde derivative (2) prepared in Example 1-2 of the present invention.
  • FIG. 3 is a diagram showing a 1 H-NMR spectrum of the QCA prepared in Examples 1-4 of the present invention.
  • Example 4 is [4- (1,3,2-dioxaborinan-2-yl) phenyl] methyl N- (5-methyl-2-styryl-1,3-dioxan- prepared in Example 3-4 of the present invention. It is a diagram showing the 1 H-NMR spectrum of 5-yl) carbamate.
  • Figure 5 is a diagram showing the 1 H-NMR spectrum after the hydrolysis reaction in cancer cells of QCA prepared in Examples 1-4 of the present invention.
  • FIG. 6 is a diagram showing the results of measuring the sensitivity of QCA to H 2 O 2 by chemiluminescence.
  • FIG. 7 is a view showing the results of measuring the change in the level of GSH when the QCA of the present invention in the cell.
  • Figure 8 is a view showing the results of measuring the generation of free radicals when treating the QCA of the present invention in cells.
  • Figure 9 is a diagram showing the results of measuring cytotoxicity when the intracellular treatment of the QCA of the present invention.
  • FIG. 10 is a diagram showing the results of measuring cytotoxicity when the intracellular QBCA treatment of the present invention.
  • Figure 11 shows the results of Western blot analysis when the QCA of the present invention in DU145 cells.
  • Figure 12 shows the results of Western blot analysis when the QCA of the present invention in SW620 cells.
  • Figure 13 is a diagram showing the results of electrophoresis analysis of the segment of DNA of SW620 cells treated with QCA of the present invention.
  • Figure 14 is a diagram showing the results of analyzing the DNA segment of SW620 cells treated with QCA of the present invention over time.
  • Figure 15 is a diagram showing the flow cytometry results of DU145 cells treated with QCA of the present invention.
  • Figure 16 shows the flow cytometry results of SW620 cells treated with QCA of the present invention.
  • Figure 17 is a diagram showing the results of measuring the tumor size of xenograft model according to administration of the QCA of the present invention.
  • Figure 18 is a diagram showing the results of measuring the tumor size of a xenograft model according to the dose of the QCA of the present invention.
  • FIG. 19 is a diagram showing the results of analyzing the tumor lysate of the mice administered QCA of the present invention by LC-MS / MS.
  • FIG. 20 is a diagram confirming the cell death through H & E staining of SW620 cells treated with QCA of the present invention.
  • Figure 21 is a diagram confirming the cell death through TUNEL staining of SW620 cells treated with QCA of the present invention.
  • FIG. 22 is a diagram confirming whether the organ damage through the H & E staining of liver and heart tissue of the mouse administered the QCA of the present invention.
  • Figure 23 is a diagram confirming whether the organ damage through the TUNEL staining of liver and heart tissue of the mice administered the QCA of the present invention.
  • 24 is a diagram confirming the toxicity of the QCA through the ALT evaluation of the mice administered the QCA of the present invention.
  • 25 is a diagram confirming the toxicity of the QCA through H & E staining of liver and heart tissue of the mouse administered the QCA of the present invention.
  • the present invention provides a hybrid anticancer prodrug represented by the following Chemical Formula 1 as a hybrid anticancer prodrug which simultaneously generates cinnam aldehyde and quinone methide.
  • R is H-, HO-, CH 3 O- or C 6 H 5 COO-, R 'is H- or CH 3- , X is O or NH.
  • cinnamaldehyde is known to induce apoptosis through the generation of reactive oxygen species (ROS), but has weak cytotoxicity to normal cells.
  • ROS reactive oxygen species
  • the utilization of cinnamic aldehyde has been limited by its short half-life in blood and low activity compared to common anticancer drugs.
  • the anticancer prodrug is composed of a boronate compound moiety and a cinnamic aldehyde derivative moiety that produces quinone methoxide, and the boronate compound moiety and a cinnamic aldehyde derivative moiety are carbonate (carbonate, -OCOO-) or carbamate. , -NHCOO-) can be connected.
  • the boronate compound moiety must be linked with -COO-.
  • the quinone metes and cinnamic aldehydes can be produced by hydrogen peroxide (H 2 O 2 ) and acidic pH, in particular they can be produced specifically in cancer cells.
  • quinone methoxide is produced through boronate oxidation by hydrogen peroxide, and the quinone methoxide produced may be alkylated with antioxidant glutathione (GSH) to inhibit the antioxidant system and increase oxidative stress.
  • GSH antioxidant glutathione
  • the acidic pH may cleave the acetal bond of the present invention to release cinnamic aldehyde, wherein the released cinnamic aldehyde may generate ROS to promote apoptosis.
  • ROS produced by the release of cinnamic aldehyde is accumulated in large amounts to further promote apoptosis.
  • the hybrid anticancer drugs of the present invention exhibit synergistic anticancer effects through a dual stimulation response and sequential therapeutic action in a cancer cell-specific manner.
  • step (c) reacting the cinnamic aldehyde derivative prepared in step (b) with carbonyldiimidazole to prepare a cinnamic aldehyde emitting compound
  • step (d) reacting the boronate compound prepared in step (a) and the cinnamic aldehyde emitting compound prepared in step (c); provides a method for producing a hybrid anticancer prodrug .
  • R is as defined in Formula 1.
  • Step (a) is a step of preparing a boronate compound to produce a quinone methide, by reacting the diol compound and boronic acid compound in an organic solvent to prepare a boronate compound.
  • the diol compound is preferably but not limited to propanediol, butanediol, pentanediol, boronic acid compound is (4-hydroxymethylphenyl) boronic acid ((4-Hydroxymethylphenyl) boronic acid), (4- (hydroxymethyl) 2-methylphenyl) boronic acid ((4- (hydroxymethyl) -2-methylphenyl) boronic acid) and (2-fluoro-4- (hydroxymethyl) phenyl) boronic acid ((2-fluoro-4- (hydroxymethyl phenyl) boronic acid) is preferred.
  • boronic acid compound is (4-hydroxymethylphenyl) boronic acid ((4-Hydroxymethylphenyl) boronic acid), (4- (hydroxymethyl) 2-methylphenyl) boronic acid ((4- (hydroxymethyl) -2-methylphenyl) boronic acid) and (2-fluoro-4- (hydroxymethyl) phenyl) boronic acid ((2-
  • the step (b) is to prepare a cinnamic aldehyde derivative having an acetal bond, cinnamic aldehyde is added to the acidic solution and the reaction is carried out at a high temperature of 70 ⁇ 100 °C and then the solvent is evaporated.
  • the acidic solution is preferably p-toluene sulfonic acid or sulfuric acid, but is not limited thereto.
  • the prepared cinnamic aldehyde derivative is decomposable in acid, including an acetal bond, thereby enabling the release of cinnamic aldehyde in cancer cells.
  • step (c) is to prepare a cinnamic aldehyde emitting compound, the cinnamic aldehyde derivative prepared in step (b) is dissolved in an organic solvent with carbonyldiimidazole and reacted at 20 ⁇ 40 °C and evaporated the solvent Get
  • Step (d) is a step of preparing a hybrid anticancer prodrug which simultaneously produces cinnamic aldehyde and quinone metide, the boronate compound prepared in step (a) and the cinnamic aldehyde emitting compound prepared in step (c) Is reacted in an organic solvent and then the solvent is evaporated.
  • the organic solvent used in each step may include tetrahydrofuran, dichloromethane, hexane, dioxane, benzene, dimethyl sulfoxide, dimethylformamide, and the like, but is not limited thereto.
  • step (c) reacting the boronate compound prepared in step (a) with carbonyldiimidazole to prepare a quinone metide releasing compound;
  • step (d) reacting the cinnamic aldehyde compound prepared in step (b) with the quinone methoxide releasing compound prepared in step (c).
  • R is as defined in Formula 1.
  • the present invention also provides a composition for the prevention or treatment of cancer comprising the hybrid anticancer prodrug as an active ingredient.
  • the composition includes a pharmaceutical composition and a food composition.
  • the hybrid anticancer prodrug of the present invention is to release the quinone meth and cinnamaldehyde sequentially by H 2 O 2 and acidic pH to alkylate the antioxidant GSH by the release of quinone methion to inhibit the antioxidant system and oxidize May increase stress
  • a large amount of free radicals (ROS) produced by the release of cinnamic aldehyde promotes apoptosis, thereby causing a synergistic anticancer effect through a dual stimulation response and a sequential therapeutic action in a cancer cell-specific manner It can be usefully used as an anticancer agent.
  • ROS free radicals
  • the cancer includes lung cancer, pancreatic cancer, colon cancer, colorectal cancer, myeloid leukemia, thyroid cancer, myeloid dysplasia syndrome (MDS), bladder carcinoma, epidermal carcinoma, melanoma, breast cancer, prostate cancer, head and neck cancer, uterine cancer, ovarian cancer, brain cancer , Gastric cancer, laryngeal cancer, esophageal cancer, bladder cancer, oral cancer, cancer of mesenchymal origin, sarcoma, teratoma, neuroblastoma, renal carcinoma, liver cancer, non-Hodgkin's lymphoma, multiple myeloma, and thyroid undifferentiated cancer Can be.
  • MDS myeloid dysplasia syndrome
  • composition of the present invention may further contain at least one known effective ingredient having the effect of preventing or treating cancer in combination with the hybrid anticancer prodrug.
  • composition of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions. It may also be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions according to conventional methods. Suitable formulations known in the art are preferably those disclosed in Remington's Pharmaceutical Science, recently, Mack Publishing Company, Easton PA.
  • Carriers, excipients and diluents that may be included include 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, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil.
  • diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, lactose, It is prepared by mixing gelatin.
  • excipients such as starch, calcium carbonate, sucrose, lactose, It is prepared by mixing gelatin.
  • lubricants such as magnesium stearate and talc are also used.
  • Oral liquid preparations include suspensions, solvents, emulsions, and syrups, 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.
  • 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.
  • base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
  • the term "administration" means providing a subject with any of the compositions of the present invention in any suitable manner.
  • Preferred dosages of the pharmaceutical compositions of the present invention vary depending on the condition and weight of the individual, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art.
  • the anticancer prodrugs of the present invention may be administered in an amount of 1 mg / kg to 10000 mg / kg per day, and may be administered once or several times a day.
  • composition of the present invention can be administered to a subject by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections.
  • composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of cancer.
  • the food composition of the present invention may be used as it is, or may be used in combination with other food or food ingredients, or may be appropriately used according to a conventional method.
  • the mixed amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment).
  • the compositions of the present invention are added in an amount of up to 15% by weight, preferably up to 10% by weight relative to the raw materials.
  • the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.
  • foods to which the substance may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gum, ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.
  • the health beverage composition of the present invention may include various flavors or natural carbohydrates, and the like as an additional ingredient, as in a general beverage.
  • the natural carbohydrates described above may be used as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, and synthetic sweeteners such as saccharin and aspartame.
  • the proportion of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.
  • the composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, It may include a carbonation agent used in the carbonated beverage.
  • the composition of the present invention may include a pulp for the production of natural fruit juice, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
  • 1,1'-Carbonyldiimidazole (4.1 g) and cinnamic aldehyde derivative (2) (3.0 g) prepared in 1-2 were dissolved in 50 mL of dry dichloromethane. After reacting at room temperature for 30 minutes. The reaction mixture was evaporated under reduced pressure to remove dichloromethane and purified by column chromatography using ethyl acetate as elution solvent to afford the title compound (3).
  • the title compound (2) was prepared in the same manner as in Example 1-2, except that benzoyloxycinnamaldehyde was used instead of cinnamic aldehyde in Example 1-2.
  • Example 1-3 in Example 1-3 except for using the cinnamic aldehyde derivative (2) prepared in Example 2-2 instead of the cinnamic aldehyde derivative (2) prepared in Example 1-2 In the same manner as in the title compound (3) was prepared.
  • Example 1-4 except that the cinnamic aldehyde emitting compound (3) prepared in Example 2-3 was used instead of the cinnamic aldehyde emitting compound (3) prepared in Examples 1-3.
  • the title compound (QBCA) was prepared.
  • N- (1,3-dihydroxy-2-methylpropan-2-yl) -2,2,2-trifluoroacetamide (4 g) was added thereto with ethyl acetate, and the mixture was diluted with THF at 70 ° C. for 6 hours.
  • 1,1'-Carbonyldiimidazole (4.1 g) and the boronate compound (1) (3.0 g) prepared in 3-1 above were dissolved in 50 mL of dry dichloromethane. After reacting at room temperature for 30 minutes. The reaction mixture was evaporated under reduced pressure to remove dichloromethane and purified by column chromatography using ethyl acetate as elution solvent to afford the title compound (3).
  • the peak of 4.1ppm confirmed the successful binding of boronate and cinnamic aldehyde.
  • QCA was reacted in a diphenyl oxalate solution containing rubrene, and then evaluated by measuring chemiluminescence intensity. Specifically, QCA was added to H 2 O 2 solution (1 ⁇ M) and reacted for 1 minute. Diphenyl oxalate solution was then added to the H 2 O 2 solution and chemiluminescence intensity was measured using a luminometer (Femtomaster FB 12, Zylux Corporation, TN, US). The results are shown in FIG.
  • the untreated H 2 O 2 solution showed a significantly high luminescence intensity (1.1 ⁇ 10 5 RLU), but it was confirmed that the chemiluminescence intensity decreased considerably when QCA was added. This indicates that in QCA the QCA is activated such that the boronate ester reacts particularly rapidly with H 2 O 2 to release QM.
  • Prostate cancer cell line (DU145), colon cancer cell line (SW620) and fibroblast line (NIH3T3) each were seeded in 6 well plates (5 ⁇ 10 5 / well) to reach 80% confluency. After treatment with various compounds at 37 ° C. for 1.5 hours, cells were harvested and washed with PBS. It was then dissolved in ice in 40 ⁇ L of Triton Triton X-100 (X-100) lysis buffer. After 20 minutes, the lysates were centrifuged at 9,000 g and 10 ⁇ L of supernatant was mixed with 50 ⁇ L of Ellman's reagent (0.5 mM DTNB).
  • the amount of GSH was quantified by measuring absorbance at 405 nm using a microplate reader (Biotek Instruments, Winooski, VT). The percentage of GSH content in the treated cells was compared with the basal GSH content measured in untreated cells and the results are shown in FIG. 7.
  • Prostate cancer cell lines (DU145) (4 ⁇ 10 5 ) were inoculated into 24 well culture plates, followed by incubation for 24 hours and treatment with cinnamic aldehyde or QCA for 12 hours. Cells were washed twice with fresh medium and resuspended in 1 ⁇ binding buffer at a concentration of 1 ⁇ 10 5 cells / mL. The cell suspension (100 ⁇ L) was transferred to a 5 mL culture tube and mixed gently by addition of 5 ⁇ M DCFH-DA (2 ′, 7′-dichlorofluorescein diacetate). Cells were incubated at room temperature for 15 minutes and then 400 ⁇ L of 1 ⁇ binding buffer was added. Stained cells were measured using flow cytometry (FACS Caliber, Becton Dickinson, San Jose, Calif.), And the results are shown in FIG. 8. A total of 1.5 ⁇ 10 4 cells were analyzed for each sample.
  • the cytotoxicity of QCA prepared in Example 1 and QBCA prepared in Example 2 was measured by MTT assay. Specifically, prostate cancer cell line (DU145), colorectal cancer cell line (SW620) and fibroblast cell line (NIH3T3) were respectively inoculated in 24 well plates at a density of 1 ⁇ 10 5 cells / well and at ⁇ 90% confluency. Incubate for 24 hours to reach. Cells were treated with various amounts of QCA, QBCA, Q1 or cinnamic aldehyde and incubated for 24 hours. 100 ⁇ L of MTT solution was added to each well and incubated for 4 hours. The resulting formazan crystals were dissolved in 200 ⁇ L of dimethyl sulfoxide.
  • Prostate cancer cell line (DU145) and colorectal cancer cell line (SW620) (1 ⁇ 10 6 / well) were treated with various concentrations of cinnamic aldehyde or QCA for 8 hours and washed twice with fresh PBS. Protein was extracted from the cells using lysis buffer. Electrophoresis was performed using 20 g of cell lysate in 1% polyacrylamide gel and the protein was transferred to PVDF membrane (Millipore, Billerica, Mass.).
  • DNA fragmentation assay of SW620 was performed. First, DU145 and SW620 cells were incubated for 24 hours, followed by 100 ⁇ m of cinnamaldehyde, 100 ⁇ m of Q1 (produced 4- (1,3,2-dioxaborinan-2-yl) benzyl benzyl carbonate, quinone meted only) and 25 ⁇ m. DNA was isolated by treatment with 50 ⁇ m, 100 ⁇ m QCA, and nucleosomal DNA fragmentation was observed by electrophoresis, and the results are shown in FIG. 13.
  • DU145 and SW620 cells were incubated for 24 hours and treated with cinnamic aldehyde, Q1 or QCA for 24 hours, followed by Annexin V-FITC and markers for cell survival as markers of apoptosis.
  • Flow cytometry was performed using PI (propodium idoide) as shown in FIGS. 15 and 16.
  • QCA-treated cells showed faster apoptosis than cinnamic aldehyde and Q1 treated cells, and Annexin V and PI cells increased with increasing QCA dose.
  • tumors of a xenograft model were observed.
  • cancer cells were injected into the flank subcutaneous tissue of the mouse using a xenograft mouse model of DU145 and SW620, and administration of cinnamic aldehyde, Q1, cinnamic aldehyde and Q1, or QCA began when the tumor began to touch small after cell transplantation.
  • the tumor size of the mouse was measured, the results are shown in Figure 17.
  • mice treated with QCA a significantly smaller tumor size was confirmed as compared to the case of the combination of cinnamic aldehyde, Q1 or cinnamic aldehyde and Q1.
  • the best effect for mice treated with QCA indicates that QCA can release QM and cinnamaldehyde continuously.
  • Tumor lysate was analyzed by LC-MS after 22 days to confirm the production of QM and cinnamic aldehyde in the tumor, and the results are shown in FIG. 19.
  • the SW620 tumor mouse model was subjected to H & E staining of tumors after administration of cinnamic aldehyde, Q1, cinnamic aldehyde and Q1, or QCA once every three days for 21 days. The results are shown in FIG. 20.
  • TUNEL terminal deoxynucleotidyl transferase dUTP nick end labeling
  • mice were injected with QCA (2 mg / kg) once every 2 days for 10 days, followed by ALT (alanine transaminase) evaluation and H & E staining of liver and heart tissues. It is shown in 24 and 25.
  • the above ingredients are mixed and filled in an airtight cloth to prepare a powder.
  • tablets are prepared by tableting according to a conventional method for preparing tablets.
  • the above ingredients are mixed and filled into gelatin capsules to prepare capsules.
  • the amount of the above ingredient is prepared per ampoule (2 ml).
  • Vitamin B6 0.5 mg
  • composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method.
  • the granules may be prepared and used for preparing a health food composition according to a conventional method.

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Abstract

La présente invention concerne un promédicament hybride anticancéreux produisant simultanément le cinnamaldéhyde et le méthide de quinone. Le promédicament hybride anticancéreux, selon la présente invention, libère séquentiellement le méthide de quinone et le cinnamaldéhyde par H2O2 et le PH acide, et donc produit l'alkylation de GSH antioxydant par la libération du méthide de quinone, ce qui inhibe un système anti-oxydatif et augmente le stress oxydatif, et génère et accumule l'espèce d'oxygène réactif (ROS) par la libération du cinnamaldéhyde, ce qui favorise l'apoptose et, par conséquent, le promédicament hybride anticancéreux selon la présente invention peut être favorablement utilisé comme médicament anticancéreux par la création d'un effet anticancéreux synergique par réponse à un double stimulus et l'action thérapeutique séquentielle à la manière spécifique de cellules cancéreuses.
PCT/KR2015/000067 2014-01-03 2015-01-05 Promédicament hybride anticancéreux produisant simultanément le cinnamaldéhyde et le méthide de quinone et son procédé de préparation Ceased WO2015102448A1 (fr)

Priority Applications (1)

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US15/109,621 US9950071B2 (en) 2014-01-03 2015-01-05 Hybrid anticancer prodrug simultaneously producing cinnamaldehyde and quinone methide and method for preparing same

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KR20140000653 2014-01-03
KR10-2014-0000653 2014-01-03
KR10-2015-0000660 2015-01-05
KR1020150000660A KR101688887B1 (ko) 2014-01-03 2015-01-05 신남알데히드 및 퀴논 메티드를 동시에 생성하는 혼성 항암 전구약물 및 이의 제조방법

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016073357A1 (fr) * 2014-11-03 2016-05-12 Beth Israel Deaconess Medical Center, Inc. Nouveaux composés anti-oxydants activables par peroxyde d'hydrogène, composés anti-oxydants et procédés les utilisant

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000033858A1 (fr) * 1998-12-07 2000-06-15 Ecosmart Technologies, Inc. Composition anticancereuse et procede utilisant des huiles essentielles vegetales naturelles avec des modulateurs de transduction de signal
WO2005065361A2 (fr) * 2003-12-31 2005-07-21 Khosrow Kashfi Composes et compositions destines au traitement de maladies dysplasiques, et leurs methodes d'utilisation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000033858A1 (fr) * 1998-12-07 2000-06-15 Ecosmart Technologies, Inc. Composition anticancereuse et procede utilisant des huiles essentielles vegetales naturelles avec des modulateurs de transduction de signal
WO2005065361A2 (fr) * 2003-12-31 2005-07-21 Khosrow Kashfi Composes et compositions destines au traitement de maladies dysplasiques, et leurs methodes d'utilisation

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016073357A1 (fr) * 2014-11-03 2016-05-12 Beth Israel Deaconess Medical Center, Inc. Nouveaux composés anti-oxydants activables par peroxyde d'hydrogène, composés anti-oxydants et procédés les utilisant

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