WO2017028602A1 - 亚丁基苯酞的用途 - Google Patents
亚丁基苯酞的用途 Download PDFInfo
- Publication number
- WO2017028602A1 WO2017028602A1 PCT/CN2016/084686 CN2016084686W WO2017028602A1 WO 2017028602 A1 WO2017028602 A1 WO 2017028602A1 CN 2016084686 W CN2016084686 W CN 2016084686W WO 2017028602 A1 WO2017028602 A1 WO 2017028602A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- stem cells
- cancer stem
- cancer
- cells
- cell
- Prior art date
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- 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
- A61K31/365—Lactones
-
- 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 invention relates to the application of butylidenephthalide (BP), which comprises preventing, slowing down and/or inhibiting cancer stem cells (CSCs), especially oral cancer stem cells, nasopharyngeal carcinoma stem cells, esophageal cancer stem cells, bone marrow Tumor stem cells, skin cancer stem cells, melanoma stem cells, thyroid cancer stem cells, lymphoma stem cells, blood cancer stem cells, breast cancer stem cells, bladder cancer stem cells, ovarian cancer stem cells, cervical cancer stem cells, prostate cancer stem cells, gastric cancer stem cells, liver cancer stem cells, lung cancer stem cells Colorectal cancer stem cells, rectal cancer stem cells, pancreatic cancer stem cells, gallbladder cancer stem cells, kidney cancer stem cells, glioblastoma stem cells, thymic malignant stem cells, rhabdomyosarcoma stem cells, brain tumor stem cells, and medulloblastoma stem cells, It is the growth, migration, invasion and/or metastasis of oral cancer stem cells, pan
- Tumor refers to the abnormal pathological changes of cells. Under the action of various carcinogenic factors, the tumors cause the cells of the local tissues of the body to lose the normal regulation of their growth at the genetic level. The cells proliferate abnormally and accumulate into masses, hence the name “tumor.” Among them, “cancer” is the most common form of tumor. The abnormally proliferating “cancer cells”, in addition to being aggregated into a mass, will spread and metastasize to other tissues or organs of the body, so it is also called malignant tumor. The proliferation and metastasis of cancer cells can cause serious physiological dysfunction and is difficult to cure, so In recent years, cancer has become the leading cause of human death worldwide.
- Traditional cancer treatment methods include surgical treatment, chemotherapy, and radiation therapy.
- the surgical removal of the mass is usually not effective in curing cancer. Probably because the cancer cells that are not completely resected are still growing sustainably, the patient's condition is gradually increased. Deteriorating. Therefore, it is generally not possible to treat cancer only by surgical resection, but with other treatments such as chemotherapy and/or radiation therapy.
- chemotherapy is the use of chemical drugs (for example, alkylating agents) to kill fast-growing cancer cells.
- drugs used in chemotherapy also act on normal cells, causing serious side effects to cancer patients, including vomiting, baldness, fatigue, bleeding, and anemia.
- radiotherapy eg. Gamma Knife Therapy
- the rapidly dividing cancer cells are sensitive to radiation compared to normal cells, causing DNA breakage of cancer cells and killing cancer cells.
- destroying cancer cells with high-energy radiation also illuminates normal cells, which can lead to side effects such as white blood cell loss, fatigue, insomnia, pain, and poor appetite.
- chemotherapy and radiation therapy are not effective in treating some advanced patients.
- cancer stem cells have stem cell properties, namely self-renewal and differentiation.
- CSCs cancer stem cells
- cancer stem cells have the potential to form tumors and develop into cancer, especially when transferred to other tissues or organs of the body, to develop into other types of cancer.
- cancer stem cells are more resistant to chemotherapy or radiation therapy in different cancer tissues such as blood cancer, breast cancer, brain cancer, ovarian cancer, prostate cancer, colorectal cancer, rectal cancer, and oral cancer.
- BP butylidenephthalide
- EMT epithelial-mesenchymal transition
- gelatinase of cancer stem cells.
- the performance of gelatinase in particular, can effectively inhibit the performance of Sox-2, Oct4, and EZH2, thereby inhibiting the growth, migration, invasion, and metastasis of cancer stem cells, thereby providing prevention, slowing, and/or inhibiting the growth of cancer stem cells.
- CSCs cancer stem cells
- BP butylidenephthalide
- BP butylene benzoquinone
- the preparation is for inhibiting the performance of Sox-2, Oct4, and EZH2 to prevent, slow down, and/or inhibit the growth, migration, invasion, and/or metastasis of cancer stem cells.
- the cancer stem cell is at least one of the following: oral cancer stem cells, nasopharyngeal carcinoma stem cells, esophageal cancer stem cells, myeloma stem cells, skin cancer stem cells, melanoma stem cells, thyroid cancer stem cells, lymphoma stem cells, blood cancer stem cells, breast cancer stem cells.
- bladder cancer stem cells bladder cancer stem cells, ovarian cancer stem cells, cervical cancer stem cells, prostate cancer stem cells, gastric cancer stem cells, liver cancer stem cells, lung cancer stem cells, colon cancer stem cells, rectal cancer stem cells, pancreatic cancer stem cells, gallbladder cancer stem cells, kidney cancer stem cells, nerve glue Maternal tumor stem cells, thymic malignant stem cells, rhabdomyosarcoma stem cells, brain tumor stem cells, and medulloblastoma stem cells.
- the cancer stem cells are oral cancer stem cells, pancreatic cancer stem cells, and/or brain tumor stem cells.
- the cancer stem cell is an oral cancer stem cell.
- the cancer stem cell is a pancreatic cancer stem cell.
- the cancer stem cell is a brain tumor stem cell.
- the preparation is a medicament, a food, or a food additive.
- the preparation is a medicament and the amount of the medicament is in the form of butylene benzoquinone (BP) It is about 30 mg/kg body weight to about 500 mg/kg body weight per day.
- BP butylene benzoquinone
- the amount of the agent is from about 40 mg/kg body weight to about 120 mg/kg body weight per day based on butylene benzoquinone (BP).
- Figure 1 is a histogram showing the relative survival rates of brain tumor stem cell CSCs CD133+ cell lines after different treatments, wherein Figure 1A shows the results after treatment with different concentrations of butylenephthalide (BP), and Figure 1B shows the different concentrations.
- BP butylenephthalide
- BCNU bischloroethyl nitrosourea
- Figure 2 is an oral cancer stem cell ALDH1 + CD44 + -1 showing normal human oral keratinocytes (Normal Human Oral Keratinocyte, NHOK, ie) or head-neck cancer-initiating cells treated with different concentrations of butylene benzoquinone (BP).
- Figure 3 to Figure 5 show the results of Western blot analysis of the stemness maintenance marker proteins of brain tumor stem cell CSCs CD133+ cell lines treated with different concentrations of butylenephthalide (BP).
- Photograph showing the expression of EZH2 protein and actin Figure 4 is a photographic diagram showing the expression of CD133 protein and actin
- Figure 5 is a graph showing the expression of Sox-2 protein, Oct4 protein, and ⁇ -actin.
- Figure 6 is a graph showing the Sox-2 protein and CD133 protein of pancreatic cancer cell MiaPaCa-2 cell line treated with different concentrations of butylene benzoquinone (BP) by western dot method. Photographs of the expression of CD44 protein and ⁇ -actin;
- Figure 7 is a diagram showing oral cancer stem cell ALDH1 + CD44 + -1 cell line and ALDH1 + CD44 + -2 cell line which were analyzed by Western blot method for different concentrations of butylated phenylhydrazine (BP)-treated head and neck cancer-derived cells. Photographs of the performance of Oct4, Sox-2, and GAPDH proteins;
- BP butylated phenylhydrazine
- Figure 8 shows the flow cytometry analysis of oral cancer stem cell ALDH1 + CD44 + -1 cell line and ALDH1 + CD44 + -2 cell line of head-neck cancer-derived cells treated with different concentrations of butylenephthalide (BP). Results obtained from ALDH activity in the presence or absence of DEAB;
- Figure 9A is a photographic diagram showing the expression of Axl protein, Gas6 protein, and actin of brain tumor stem cell CSCs CD133+ cell lines treated with different concentrations of butylenephthalide (BP) by Western blotting;
- BP butylenephthalide
- Figure 9B is a photographic diagram showing the expression of Axl gene and GAPDH gene of brain tumor stem cell CSCs CD133+ cell line treated with different concentrations of butylenephthalide (BP) by reverse transcription-polymerase chain reaction;
- BP butylenephthalide
- Figure 10 is a photographic diagram showing the expression of MMP-2 protein and actin in brain tumor stem cell CSCs CD133+ cell lines treated with different concentrations of butylenephthalide (BP) by Western blotting;
- BP butylenephthalide
- Figure 11 shows the results of an invasive assay system for analyzing the invasive ability of oral cancer stem cell ALDH1 + CD44 + -1 cell line and ALDH1 + CD44 + -2 cell line treated with different concentrations of butylenephthalide (BP),
- Figure 11A is a photographic view showing the staining results of the polycarbonate filter located in the lower chamber of the invasion test system, and
- Figure 11B is a histogram showing the relative invasive ability after quantification;
- Figure 12 is a graph showing E-cadherin protein, TGF ⁇ -1 protein, Slug protein, and ⁇ -actin in brain tumor stem cell CSCs CD133+ cell lines treated with different concentrations of butylene benzoquinone (BP) by western dot method. Photograph of performance;
- Figure 13 is a graph showing the results of testing the migration ability of brain tumor stem cell CSCs CD133+ cell lines after different treatments by a wound healing test, wherein the upper two rows show photographs of the results after treatment with different concentrations of butylenephthalide (BP). Figure, while the next two rows show photographic photographs of the results after treatment with different concentrations of bischloroethylnitrosourea (BCNU);
- BP butylenephthalide
- BCNU bischloroethylnitrosourea
- Figure 14A is a photograph showing the observation of a brain tumor stem cell CSCs CD133+ cell line carrying pcDNA3.0-Axl or pcDNA3.0-neo plastids under a microscope;
- Figure 14B is a photographic diagram showing the expression of Axl-1 protein and ⁇ -actin in a brain tumor stem cell CSCs CD133+ cell line carrying pcDNA3.0-Axl or pcDNA3.0-neo plastids by Western blotting;
- Figure 14C shows the migration ability of brain tumor stem cell CSCs CD133+ cell lines with pcDNA3.0-Axl or pcDNA3.0-neo plastids treated with different concentrations of butylenephthalide (BP) in a wound healing test. result;
- BP butylenephthalide
- Figure 15 shows the results of the tumor initiating activity of oral cancer stem cells treated with different concentrations of butylenephthalide (BP) in the form of softwood gel cell community formation, wherein Figure 15A shows ALDH1 + CD44 + -1 cells. Photograph of the cell population formation of the strain and the ALDH1 + CD44 + -2 cell line, and Figure 15B is a histogram showing the relative cell colony forming ability of the ALDH1 + CD44 + -1 cell line and the ALDH1 + CD44 + -2 cell line;
- BP butylenephthalide
- Figure 16 shows the results of an animal experiment to analyze the tumor-initiating activity of butylene benzoquinone (BP) in inhibiting cancer stem cells, including control group mice not treated with butylenephthalide (BP) ( Show), experimental group of mice injected with 100 mg / kg body weight of butylene benzoquinone (BP) daily (to Show), and experimental mice injected with 200 mg/kg body weight of butylene benzoquinone (BP) daily (to The results are shown, wherein Fig. 16A shows the average body weight (gram) of each group of mice at different time points, Fig. 16B shows the average tumor volume of each group of mice at different time points, and Fig. 16C shows the GFP of each group of mice. Photograph of signal intensity, and Fig. 16D is a histogram showing the results of quantifying the GFP signal intensity of each group of mice.
- Fig. 16A shows the average body weight (gram) of each group of mice at different time points
- Fig. 16B shows the average tumor volume of each group of
- a therapeutically effective amount refers to an amount of a compound that is effective to at least partially improve the condition of a suspected individual when administered to an individual; the term "individual” refers to a mammal, which may be a human or a non-human animal.
- butylene benzoquinone has at least one of the following effects: inhibiting the survival rate of cancer stem cells, inhibiting the expression of cancer stem cell growth factor (for example, EZH2 protein), and inhibiting the stem cell of cancer stem cells.
- markers eg, CD133 protein, Sox-2 protein, Oct4 protein
- markers eg, CD133 protein, Sox-2 protein, Oct4 protein
- inhibition of genes associated with cancer stem cell proliferation, anti-apoptosis, migration, and invasion eg Such as: Axl gene
- protein for example: Axl protein, Gas6 protein, MMP-2 protein
- inhibition of proteins associated with cancer stem cell metastasis eg: E-cadherin protein, TGF ⁇ -1 protein, and Slug protein
- Performance and the ability to inhibit the migration of cancer stem cells to the wound.
- butylene benzoquinone (BP) prevents, slows, and/or inhibits the growth, migration, invasion, and/or metastasis of cancer stem cells by inhibiting the performance of Sox-2, Oct4, and EZH2.
- the present invention relates to the use of butylene benzoquinone (BP) for the manufacture of a formulation.
- the preparation is for preventing, slowing down and/or inhibiting the growth, migration, invasion and/or metastasis of cancer stem cells.
- the preparation of the present invention can be applied to any suitable cancer stem cells, including, for example, oral cancer stem cells, nasopharyngeal carcinoma stem cells, esophageal cancer stem cells, myeloma stem cells, skin cancer stem cells, melanoma stem cells, thyroid cancer stem cells, lymphoma stem cells, blood cancer stem cells.
- suitable cancer stem cells including, for example, oral cancer stem cells, nasopharyngeal carcinoma stem cells, esophageal cancer stem cells, myeloma stem cells, skin cancer stem cells, melanoma stem cells, thyroid cancer stem cells, lymphoma stem cells, blood cancer stem cells.
- the formulation is for preventing, slowing, and/or inhibiting the growth, migration, invasion, and/or metastasis of oral cancer stem cells, pancreatic cancer stem cells, and/or brain tumor stem cells.
- the preparation provided according to the present invention may be in any suitable form without particular limitation.
- the preparation may be provided in the form of a medicament or may be provided in the form of a solid or fluid food or food additive.
- the medicament may be in a correspondingly suitable dosage form depending on the desired administration form.
- the agent can be administered orally or non-orally (eg, subcutaneous, intravenous, intramuscular, intraperitoneal, or nasal) to a subject in need thereof.
- a suitable carrier may be employed to provide the agent.
- the agent provided by the present invention may contain any pharmaceutically acceptable carrier which does not adversely affect the desired effect of butylenephthalide (BP), such as a solvent.
- a solvent water, saline, dextrose, glycerin, ethanol or the like, and combinations thereof
- oily solvents water, saline, dextrose, glycerin, ethanol or the like, and combinations thereof
- diluents stabilizers
- absorption delaying agents disintegrating agents, emulsifiers, antioxidants, binders
- the agent may be provided in a dosage form suitable for oral administration by any suitable method, for example, a tablet (for example, a sugar-coated tablet), a pill, a capsule, a granule, a powder, a flow extract, a solution, a syrup, a suspension. Agents, emulsions, and tinctures.
- an injection form or a drip form suitable for subcutaneous, intravenous, intramuscular, or intraperitoneal injection one or more kinds of, for example, a solution, a salt buffer (such as a phosphate buffer or a lemon) may be contained in the agent provided by the present invention.
- Ingredients such as acid salt buffer, solubilizer, emulsifier, 5% sugar solution, and other carriers, such as intravenous infusion, emulsion intravenous infusion, dry powder injection, suspension injection, or dry powder suspension injection
- the agent is provided.
- the medicament is prepared as a pre-injection solid
- the pre-injection solid is provided in a dosage form soluble in other solutions or suspensions, or an emulsifiable dosage form, and administered to the Prior to the needy individual, the pre-injection solid is dissolved in or emulsified to provide the desired injectable preparation.
- external preparations suitable for nasal administration or transdermal administration are, for example, emulsions, creams, gels (for example hydrogels), creams (for example, dispersion creams, ointments), sprays, or solutions (for example, washing). Liquid, suspension).
- the agent provided by the present invention may additionally contain an appropriate amount of an additive, for example, a flavoring agent, a toner, a coloring agent, etc., which can improve the mouthfeel and visual sensation of the drug during administration, and can be improved.
- an additive for example, a flavoring agent, a toner, a coloring agent, etc.
- a buffer, a preservative, a preservative, an antibacterial agent, an antifungal agent, and the like for stability and storage of the drug.
- the agent may additionally contain one or more other active ingredients, or may be used in combination with a drug containing the one or more other active ingredients to further enhance the efficacy of the agent or increase the flexibility of use of the formulation.
- the degree of blending is as long as the other active ingredient does not adversely affect the desired effect of butylenephthalide (BP).
- the agent provided by the present invention may be administered at different frequencies, such as once a day, multiple times a day, or several times a day, depending on the age, weight, and condition of the individual to be administered (eg, the condition to be treated and the condition)
- the severity varies.
- the amount is about 30 per day based on butylene benzoquinone (BP).
- mg/kg body weight refers to the amount of drug required per kilogram of body weight.
- the amount can be increased according to actual needs, for example, increased to several times or tens of times.
- the agent of the invention may be used in combination with one of the following to prevent, slow down and/or inhibit the growth, migration, invasion and/or metastasis of cancer stem cells: surgical treatment, chemotherapy, radiation therapy, antibody therapy, Immunotherapy, anti-angiogenic therapy, phenotypic altered therapy, and differentiation therapy.
- the preparation provided according to the present invention is provided in the form of a food additive, it may be in various forms such as powder, liquid, suspension, granules, etc., to facilitate the addition during the food manufacturing process; and when the preparation is food-based
- the form it may be, for example, dairy products, processed meat products, breads, noodles, biscuits, buns, juices, teas, sports drinks, nutritional drinks, etc., and may be healthy foods (for example: surgery) Post-nutrition supplements), but not limited to this.
- the health foods provided by the present invention may be consumed at different frequencies, such as once a day, multiple times a day, or once a few days, depending on the age, weight, and health condition of the individual.
- the content of butylene benzoquinone (BP) in the health food provided by the present invention can also be adjusted for a specific ethnic group, preferably adjusted to the amount to be taken daily. For example, if a recommended intake of a body is about 50 mg of butyl benzophenone (BP) per day, and the health food contains 25 mg of butylene benzoquinone (BP) per serving, the individual is eaten daily. About two servings of this healthy food.
- the recommended dosage of the health food of the present invention may be used to facilitate the user without a physician, pharmacist or Under the guidance of the relevant deacons, they can take it at home without safety concerns.
- the invention further provides a method for preventing, slowing down and/or inhibiting the growth of cancer stem cells
- a method of growth, migration, invasion, and/or metastasis comprising administering an effective amount of butylene benzoquinone (BP) to an individual in need thereof.
- BP butylene benzoquinone
- the administration form and the applicable dose of butylene benzoquinone (BP), and the aspect of the applicable cancer stem cells are as described above.
- BP butylene benzoquinone
- different concentrations (0, 25, 50, 62.5, 75, 100 ⁇ g / ml) of butylene benzoquinone (BP)
- different concentrations (0 , 25, 125, 250, 500, 1000 micromolar concentrations of bis-chloroethylnitrosourea (BCNU; one of the currently available chemotherapy drugs for the treatment of malignant glioma)
- BCNU bis-chloroethylnitrosourea
- TTZ micromolar temozolomide
- brain tumor stem cell CSCs CD133+ The cell line or the malignant glioma DBTRG cell line was treated for 24 or 48 hours, after which the brain cell stem cell CSCs CD133+ cell line and the malignant glioma DBTRG cell line were analyzed by cell viability assay (MTT assay). Survival rate.
- MTT assay cell viability assay
- the relative survival rates of brain tumor stem cell CSCs and CD133+ cell lines were significantly increased with the concentration of butylenephthalide (BP) or dichloroethylnitrosourea (BCNU) regardless of the treatment for 24 or 48 hours. And falling.
- BP butylenephthalide
- BCNU dichloroethylnitrosourea
- BP butylenephthalide
- BCNU dichloroethylnitrosourea
- TMZ temozolomide
- the half maximal inhibitory concentration (IC 50 ) is 406 micromolar (ie, 76.5 micrograms per milliliter), 377.6 micromolar (ie, 80.8 micrograms per milliliter), and greater than 1600 micromolar (ie, greater than 310.6 micrograms per milliliter, respectively).
- BP butylene benzoquinone
- BP butylene benzoquinone
- ALDH1 + CD44 + -1 and ALDH1 + of the head and neck cancer-initiating cells The survival rate of CD44 + -2 cell lines was significantly decreased with the increase of the concentration of butylene benzoquinone (BP).
- BP butylene phenyl hydrazine
- the enhancer of zeste homolog 2 (EZH2) gene is known to be a key growth factor for cancer stem cells, which affects the growth of cancer stem cells. Therefore, brain tumor stem cell CSCs CD133+ cells were treated with different concentrations (0, 12.5, 25, 50, 62.5, 75 ⁇ g/ml) of butylene benzoquinone (BP), and then the protein of the cells was extracted. The expression of EZH2 protein of brain tumor stem cell CSCs CD133+ cell line treated with butylenephthalide (BP) was confirmed by Western Blotting. The results are shown in Fig. 3.
- Example 3 Effect of butylene benzoquinone (BP) on inhibiting stem cells of cancer cells and cancer stem cells
- CD44 protein, CD133 protein, Sox-2 (Sex)-related high-mobility group box 2 protein, and Oct4 (octamer binding transcription factor 4) protein are cancer stem cells to maintain their stem cell properties ( The important marker of stemness is involved in regulating the self-renewal and differentiation of cancer stem cells. Therefore, brain tumor stem cell CSCs CD133+ cells were treated with different concentrations (0, 12.5, 25, 50, 62.5, 75 ⁇ g/ml) of butylene benzoquinone (BP) for 24 hours, followed by extraction. The protein of the cells was confirmed by Western blotting method. The CD133 protein, Sox-2 protein and Oct4 protein of brain cancer stem cell CSCs CD133+ cells treated with butylenephthalide (BP) were shown in Fig.
- BP butylenephthalide
- pancreatic cancer cell MiaPaCa-2 cell line was treated with different concentrations (0, 12.5, 25, 50 ⁇ g/ml) of butylene benzoquinone (BP) for 24 hours, after which the cells were extracted.
- BP butylene benzoquinone
- Protein, and the expression of Sox-2 protein, CD133 protein, and CD44 protein of pancreatic cancer cell MiaPaCa-2 cell line treated with butylenephthalide (BP) were confirmed by Western blotting method, and the results are shown in Fig. 6.
- oral cancer stem cells ALDH1 + CD44 + -1 or ALDH1 + CD44 + -2 cells of head-neck cancer-initiating cells were prepared at different concentrations (0, 25, 50 ⁇ g/ml) of butylenephthalide (BP).
- the strain was treated for 24 hours, after which the protein of the cells was extracted, and the oral cancer stem cell ALDH1 + CD44 + -1 of the head-neck cancer-derived cells treated with butylenephthalide (BP) was confirmed by Western blotting method and The performance of Sox-2 protein and Oct4 protein of ALDH1 + CD44 + -2 cell line is shown in Fig. 7.
- BP butylenephthalide
- ALDH protein is also an important marker for cancer stem cells to maintain their stemness and is involved in regulating the self-renewal and differentiation of cancer stem cells. Therefore, oral cancer stem cells ALDH1 + CD44 + -1 and ALDH1 + CD44 + -2 cells of head-neck cancer-initiating cells were prepared at different concentrations (0, 25, 50 ⁇ g/ml) of butylenephthalide (BP). The plants were treated for 24 hours. Thereafter, 1 ⁇ 10 5 cells from each group were suspended in a buffer solution of a 50 ⁇ l dry cell identification group (ALDEFLUOR array kit, purchased from STEMCELL Technologies, Vancouver, Canada), and ALDEFLUOR was added to the cell suspension to the final. The concentration is 1 micromolar.
- ALDEFLUOR array kit purchased from STEMCELL Technologies, Vancouver, Canada
- Example 4 Effect of butylene benzoquinone (BP) on inhibiting proliferation, anti-apoptosis, migration and invasion of cancer stem cells
- BP butylene benzoquinone
- Axl i.e., a receptor tyrosine kinase
- BP butylene benzoquinone
- MMP matrix metalloproteinase
- This study further uses a penetrating invasion test system ( System, purchased from Corning, UK), with a pore size of 8 ⁇ m polycarbonate filter (purchased from Corning, UK) to investigate the effect of butylenephthalide (BP) on inhibiting the migration and invasion of cancer stem cells.
- System purchased from Corning, UK
- BP butylenephthalide
- the base gel (Matrigel TM, available from the American company BD Pharmingen) was coated on the polycarbonate membrane was then coated polycarbonate filters placed in the transmissive Invasion Assay System In a lower chamber, a culture solution containing 10% serum was added to the lower chamber.
- oral cancer stem cells ALDH1 + CD44 + -1 and ALDH1 + CD44 + -2 against head-neck cancer-initiating cells at different concentrations (0, 25, 50 ⁇ g/ml) of butylenephthalide (BP) The cell line was processed for 24 hours. Thereafter, the treated oral cancer stem cells were further cultured in a serum-free medium at a cell density of 1 ⁇ 10 5 cells/100 ⁇ l in an upper chamber of the penetrating invasion test system. In, it lasted 24 hours.
- the culture solution in the lower chamber was removed, the polycarbonate filter in the lower chamber was taken out and fixed with a 4% aqueous formaldehyde solution, and after being stained with crystal violet, under a microscope at a magnification of 100 times.
- the number of cells in five different fields of view was taken and photographed, and the results are shown in Fig. 11A.
- the relative invasive ability of the cells of each experimental group was calculated based on the results of the treatment group at a concentration of 0 ⁇ g/ml, and the results are shown in Fig. 11B.
- BP butylene benzoquinone
- EMT epithelial-mesenchymal transition
- BP butylene benzoquinone
- TGF ⁇ -1 protein and Slug protein decreased significantly with the increase of the concentration of butylene benzoquinone (BP), while the expression of E-cadherin protein correlated with the concentration of butylene benzoquinone (BP). Increase and increase.
- BP butylene benzoquinone
- BP butylenephthalide
- BP butylene benzoquinone
- BP butylene benzoquinone
- % (v/v) fetal calf serum (Fetal Calf Serum, FCS), and 0.3% (w/v) acacia gum purchased from Sigma-Aldrich.
- FCS Fetal Calf Serum
- acacia gum purchased from Sigma-Aldrich.
- 2 ml of the above-mentioned groups of upper layer of acacia (containing oral cancer stem cells) were added to different wells after coagulation. Thereafter, the plate was placed in a 37 ° C incubator for cultivation for 4 weeks.
- Fig. 15A the number of cell colonies in the lower layer of acacia gel is significantly decreased as the concentration of butylene benzoquinone (BP) is increased; as can be seen from Fig. 15B, the relative cell colony forming ability of the cancer stem cell cells is also related to the butylene group.
- the concentration of phenylhydrazine (BP) is increased and decreased.
- BP butylenephthalide
- mice After the tumor was grown to a volume of 100 mm 3 , the mice were divided into a control group and two experimental groups (three groups, three in each group), and the mice in the two experimental groups were injected with 100 mg/kg body weight respectively. /butylidene benzoquinone (BP) per day and 200 mg / kg body weight / day, only vehicle (without butyrene benzoquinone (BP)) was administered to the control group for 6 days. Next, on the 10th day after the injection of butylenephthalide (BP) or vehicle, the average body weight of each group of mice was measured every 2 days, and the average length and average width of the tumor were measured, and the average volume of the tumor was calculated by the following formula 1.
- BP butylenephthalide
- Equation 1 [length ⁇ width 2 ] / 2 (unit: cubic millimeter)
- butylene benzoquinone can effectively inhibit the survival rate of cancer stem cells, inhibit the expression of cancer stem cell growth factor, and inhibit cancer stem cells.
- Expression of a stem cell maintenance marker, inhibition of expression of genes or proteins associated with proliferation, anti-apoptosis, migration, and invasion of cancer stem cells, inhibition of expression of genes or proteins associated with metastasis of cancer stem cells, inhibition and cancer stem cells The ability to migrate to the wound, particularly to effectively inhibit the performance of Sox-2, Oct4, and EZH2, can be used to prevent, slow, and/or inhibit the growth, migration, invasion, and/or metastasis of cancer stem cells.
Abstract
Description
Claims (10)
- 一种使用亚丁基苯酞于制造一制剂的用途,其特征在于:该制剂是用以预防、减缓及/或抑制癌症干细胞的生长、迁移、侵袭及/或转移。
- 如权利要求1所述的用途,其特征在于:该制剂用于抑制Sox-2、Oct4、及EZH2的表现以预防、减缓及/或抑制癌症干细胞的生长、迁移、侵袭及/或转移。
- 如权利要求1所述的用途,其特征在于:该癌症干细胞为以下的至少一种:口腔癌干细胞、鼻咽癌干细胞、食道癌干细胞、骨髓瘤干细胞、皮肤癌干细胞、黑色素瘤干细胞、甲状腺癌干细胞、淋巴瘤干细胞、血癌干细胞、乳癌干细胞、膀胱癌干细胞、卵巢癌干细胞、子宫颈癌干细胞、前列腺癌干细胞、胃癌干细胞、肝癌干细胞、肺癌干细胞、大肠癌干细胞、直肠癌干细胞、胰脏癌干细胞、胆囊癌干细胞、肾脏癌干细胞、神经胶母细胞瘤干细胞、胸腺恶性肿瘤干细胞、横纹肌肉瘤干细胞、脑瘤干细胞、及髓母细胞瘤干细胞。
- 如权利要求1所述的用途,其特征在于:该癌症干细胞为口腔癌干细胞、胰脏癌干细胞、及/或脑瘤干细胞。
- 如权利要求1所述的用途,其特征在于:该癌症干细胞为口腔癌干细胞。
- 如权利要求1所述的用途,其特征在于:该癌症干细胞为胰脏癌干细胞。
- 如权利要求1所述的用途,其特征在于:该癌症干细胞为脑 瘤干细胞。
- 如权利要求1至7中任一项所述的用途,其特征在于:该制剂为一药剂、一食品、或一食品添加剂。
- 如权利要求8所述的用途,其特征在于:该制剂为一药剂且该药剂的用量,以亚丁基苯酞(BP)计,为每天约30毫克/公斤体重至500毫克/公斤体重。
- 如权利要求9所述的用途,其特征在于:该药剂的用量,以亚丁基苯酞(BP)计,为每天约40毫克/公斤体重至120毫克/公斤体重。
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ES16836466T ES2789758T3 (es) | 2015-08-19 | 2016-06-03 | Uso de butilideneftalida |
JP2017513523A JP2017530113A (ja) | 2015-08-19 | 2016-06-03 | ブチリデンフタリドの使用 |
EP16836466.9A EP3338775B1 (en) | 2015-08-19 | 2016-06-03 | Use of butylidenephthalide |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201562207226P | 2015-08-19 | 2015-08-19 | |
US62/207,226 | 2015-08-19 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2017028602A1 true WO2017028602A1 (zh) | 2017-02-23 |
Family
ID=58050848
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2016/084686 WO2017028602A1 (zh) | 2015-08-19 | 2016-06-03 | 亚丁基苯酞的用途 |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP3338775B1 (zh) |
JP (1) | JP2017530113A (zh) |
ES (1) | ES2789758T3 (zh) |
TW (1) | TWI626940B (zh) |
WO (1) | WO2017028602A1 (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020011607A1 (en) | 2018-07-09 | 2020-01-16 | Fondation Asile Des Aveugles | Inhibition of prc2 subunits to treat eye disorders |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20220117929A1 (en) * | 2020-10-21 | 2022-04-21 | Buddhist Tzu Chi Medical Foundation | Method for preventing or treating high-grade serous carcinoma |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070134351A1 (en) * | 2004-10-08 | 2007-06-14 | Buddhist Tzu Chi General Hospital | Angelicae sinensis extracts useful for treatment of cancers |
CN102579434A (zh) * | 2011-01-07 | 2012-07-18 | 中国医药大学 | 治疗脑癌或用以降低脑癌细胞对替莫唑胺的抗药性的医药组合物 |
CN105030760A (zh) * | 2014-04-24 | 2015-11-11 | 长弘生物科技股份有限公司 | 稳定医药组合物 |
CN105267205A (zh) * | 2014-06-04 | 2016-01-27 | 中国医药大学 | 治疗胰脏癌的医药配方及其应用 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005082407A1 (en) * | 2003-11-03 | 2005-09-09 | Musc Foundation For Research Development | Use of quercetin and resveratrol to treat and prevent oral cancer |
TW201106947A (en) * | 2009-08-28 | 2011-03-01 | Tzyy-Wen Chiou | Anticancer formulation |
TWI413525B (zh) * | 2010-01-22 | 2013-11-01 | Univ Nat Taiwan Normal | 醫藥組成物作為製備抑制肺癌幹細胞生長之藥物之用途 |
WO2014014931A1 (en) * | 2012-07-16 | 2014-01-23 | Batra Raj K | Methods of biomarker validation and target discovery |
TWI484033B (zh) * | 2013-01-25 | 2015-05-11 | Univ China Medical | 培養幹細胞之方法及套組 |
-
2016
- 2016-06-03 WO PCT/CN2016/084686 patent/WO2017028602A1/zh active Application Filing
- 2016-06-03 TW TW105117580A patent/TWI626940B/zh active
- 2016-06-03 JP JP2017513523A patent/JP2017530113A/ja not_active Ceased
- 2016-06-03 EP EP16836466.9A patent/EP3338775B1/en active Active
- 2016-06-03 ES ES16836466T patent/ES2789758T3/es active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070134351A1 (en) * | 2004-10-08 | 2007-06-14 | Buddhist Tzu Chi General Hospital | Angelicae sinensis extracts useful for treatment of cancers |
CN102579434A (zh) * | 2011-01-07 | 2012-07-18 | 中国医药大学 | 治疗脑癌或用以降低脑癌细胞对替莫唑胺的抗药性的医药组合物 |
CN105030760A (zh) * | 2014-04-24 | 2015-11-11 | 长弘生物科技股份有限公司 | 稳定医药组合物 |
CN105267205A (zh) * | 2014-06-04 | 2016-01-27 | 中国医药大学 | 治疗胰脏癌的医药配方及其应用 |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020011607A1 (en) | 2018-07-09 | 2020-01-16 | Fondation Asile Des Aveugles | Inhibition of prc2 subunits to treat eye disorders |
Also Published As
Publication number | Publication date |
---|---|
EP3338775A4 (en) | 2019-04-03 |
ES2789758T3 (es) | 2020-10-26 |
TW201707698A (zh) | 2017-03-01 |
EP3338775A1 (en) | 2018-06-27 |
EP3338775B1 (en) | 2020-03-25 |
JP2017530113A (ja) | 2017-10-12 |
TWI626940B (zh) | 2018-06-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Singh et al. | Recent advances in pancreatic cancer: biology, treatment, and prevention | |
Liu et al. | Combined niclosamide with cisplatin inhibits epithelial-mesenchymal transition and tumor growth in cisplatin-resistant triple-negative breast cancer | |
Wu et al. | Arsenic trioxide depletes cancer stem-like cells and inhibits repopulation of neurosphere derived from glioblastoma by downregulation of Notch pathway | |
CN106466315B (zh) | 亚丁基苯酞的用途 | |
KR20200014790A (ko) | 난소암의 치료에 사용되는 티노스타무스틴 | |
US20190099398A1 (en) | Use of tetrahydrocannabinol and/or cannabidiol for increasing radiosensitivity in the treatment of a brain tumour | |
CN107683137A (zh) | 用于使用stat3途径抑制剂和激酶抑制剂治疗癌症的方法 | |
KR101747775B1 (ko) | 유포비아 인자 l1 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 함유하는 골질환의 예방 또는 치료용 약학적 조성물 | |
WO2017028602A1 (zh) | 亚丁基苯酞的用途 | |
KR102011105B1 (ko) | 고시폴 및 펜포르민을 유효성분으로 포함하는 췌장암 예방 및 치료용 약학적 조성물 | |
KR101855382B1 (ko) | 설폰아마이드계 화합물을 유효성분으로 포함하는 암의 예방, 치료 및 전이 억제용 약학적 조성물 | |
Feng et al. | Vascular disrupting effects of combretastatin A4 phosphate on murine endometriotic lesions | |
Waldt et al. | Loss of PTPRJ/DEP-1 enhances NF2/Merlin-dependent meningioma development | |
US10987339B2 (en) | Uses of butylidenephthalide | |
CN114452391A (zh) | Cdk16作为靶标在制备用于治疗三阴性乳腺癌的药物中的应用 | |
TWI531364B (zh) | γ-紅沒藥烯(γ-Bisabolene)於治療口腔癌之用途 | |
KR102249814B1 (ko) | 칼키톡신 티오아마이드 알코올 또는 이의 염을 유효성분으로 함유하는 암전이 억제용 조성물 | |
US20230310479A1 (en) | Low dose adjuvant epigenetic cancer therapy | |
CN111840412B (zh) | 茶褐素在制备抗黑色素瘤药物中的应用 | |
Bhandari et al. | Utilization of Intravenous (IV) Curcumin, Genistein, and Trastuzumab to Reduce HER2 receptors in Breast Cancer Patients | |
KR101844816B1 (ko) | 구아나벤즈를 포함하는 식욕부진증의 개선 또는 치료용 조성물 및 이를 이용한 방법 | |
CN106798744B (zh) | 一种增敏的卵巢癌治疗药物组合物及其用途 | |
US9446019B2 (en) | Sparstolonin B based pharmaceutical agent for neuroblastoma treatment | |
KR20230021987A (ko) | NK1R을 표적으로 하는 siRNA를 포함하는 뇌암 치료, 개선 또는 예방용 조성물 | |
TW201529078A (zh) | 白千層萃取物及其應用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
REEP | Request for entry into the european phase |
Ref document number: 2016836466 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2016836466 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2017513523 Country of ref document: JP Kind code of ref document: A |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 16836466 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |