CN111317740B - Notch signal pathway inhibition composition and application of composition in resisting prostatic cancer bone metastasis - Google Patents
Notch signal pathway inhibition composition and application of composition in resisting prostatic cancer bone metastasis Download PDFInfo
- Publication number
- CN111317740B CN111317740B CN202010219733.XA CN202010219733A CN111317740B CN 111317740 B CN111317740 B CN 111317740B CN 202010219733 A CN202010219733 A CN 202010219733A CN 111317740 B CN111317740 B CN 111317740B
- Authority
- CN
- China
- Prior art keywords
- cells
- prostate cancer
- bone metastasis
- andrographolide
- dioscin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
-
- 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
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Oncology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a Notch signal pathway inhibition composition and application of the Notch signal pathway inhibition composition in resisting prostatic cancer bone metastasis. The invention finds that the andrographolide and dioscin can play a synergistic role in combination, more efficiently inhibit notch signal channels, and further more efficiently play a role in resisting the bone metastasis of the prostate cancer, including inhibiting the proliferation, migration, invasion, adhesion and secretory expression of proteins related to metastasis of bone metastasis cells of the prostate cancer.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to application of andrographolide and dioscin in coordination in Notch signal path inhibition and prostate cancer bone metastasis resistance.
Background
Prostate cancer (PCa) is one of the most common malignancies in men worldwide and is characterized by a high incidence of bone metastases. The incidence of prostate cancer is highest in European and American countries in the world, but the incidence of prostate cancer is on the rise in our country in recent years with the increase of the life of people and the change of dietary structure. Early stage prostate cancer has no obvious clinical symptoms, is detected more than physical examination or lower urinary tract obstruction, bone pain and the like, and leads to many diagnosed patients with distant metastasis. Death of prostate cancer patients is often not the primary disease but is due to distant metastasis of tumors, particularly bone metastases. At present, palliative treatment mainly comprising androgen removal treatment is generally selected for clinically treating the prostatic cancer, but the palliative treatment cannot achieve good curative effect on the bone metastasis of the prostatic cancer, the prognosis of a patient with the bone metastasis is poor, and the average survival time is only 5-17.5 months. Therefore, it is important to find a treatment for bone metastasis of prostate cancer.
Previous studies have shown that the Notch signaling pathway has a regulatory effect on proliferation, apoptosis, differentiation, and adhesion of cells, but the regulatory effect exerted by the Notch signaling pathway is different in different cells or tissues. Inhibitors of the notch signaling pathway are sought to selectively treat diseases in which the notch signaling pathway is abnormally activated.
Disclosure of Invention
The invention aims to provide the application of the andrographolide and dioscin in synergy in resisting prostate cancer bone metastasis and inhibiting Notch signal pathways.
In some embodiments, the invention may include one or more of the following:
a notch signaling pathway inhibiting composition comprising andrographolide and dioscin.
2. The composition according to claim 1, wherein the Notch signaling pathway inhibitor composition has a function of inhibiting the expression of any one of Notch-1, NICD and Hes-1.
3. The composition according to claim 1, wherein the Notch signaling pathway inhibitor composition has a function of inhibiting the expression of any one of Notch-1, NICD and Hes-1 in bone metastasis cells of prostate cancer.
4. The composition according to item 1, wherein the molar concentration ratio of andrographolide to dioscin in the Notch signaling pathway inhibitory composition is (1.5-12): 1.
5. The composition of claim 1, wherein the dioscin includes a derivative form thereof; and/or, the andrographolide comprises a derivative form thereof.
6. A pharmaceutical composition for resisting bone metastasis of prostate cancer comprises andrographolide and dioscin.
7. The pharmaceutical composition according to item 6, wherein the pharmaceutical composition for resisting prostate cancer bone metastasis has at least one of the following functions: (1) inhibiting the proliferation of prostate cancer bone metastases; (2) inhibiting migration of prostate cancer bone metastases; (3) inhibiting invasion of prostate cancer bone metastasis cells; and (4) inhibiting the adhesion of prostate cancer bone metastasis cells.
8. The pharmaceutical composition according to item 6, wherein the molar concentration ratio of andrographolide to dioscin in the pharmaceutical composition for resisting prostate cancer bone metastasis is (1.5-12): 1.
9. The pharmaceutical composition according to item 6, wherein the pharmaceutical composition for preventing bone metastasis from prostate cancer has a function of inhibiting a Notch signaling pathway.
10. The pharmaceutical composition of claim 6, wherein the dioscin comprises a pharmaceutically acceptable salt or ester thereof; and/or, the andrographolide comprises a pharmaceutically acceptable salt or ester thereof.
Dioscorea opposita saponin (CAS: 19057-60-4, dioscin, for short, dio) is a main ingredient in Dioscoreaceae plant, and has effects of killing insects and resisting fungi such as Trichophyton mentagrophytes. However, no report about the bone metastasis resistance of prostate cancer and the inhibition of notch signal path of dioscin is found.
Andrographolide (CAS: 5508-58-7, andrographolide, the invention is called ANDRO for short) is the main effective component of natural plant andrographis paniculata, has the efficacy of removing heat and toxic materials, diminishing inflammation and relieving pain, has special curative effect on bacterial and viral upper respiratory tract infection and dysentery, and is known as natural antibiotic medicine. However, no report about the fact that the andrographolide resists the prostate cancer bone metastasis and inhibits notch signal pathways is found.
In the Notch signaling pathway, notch1 is used as a core protein of the pathway, when the Notch signaling pathway is activated, notch1 is cut by gamma secretase and NICD is generated to enter a cell nucleus to start the expression of downstream genes, and Hes-1 is an important target gene at the downstream of the Notch signaling pathway. Currently, the notch signal pathway is inhibited by FLI-06, RO4929097 and Semagacestat (LY 450139), and can be used for preventing or treating diseases with abnormally activated notch signal pathway.
Surprisingly, the application finds that andrographolide and dioscin can simultaneously inhibit the expression of key proteins and downstream genes (including Notch-1, NICD and Hes-1) of a Notch signaling pathway, and synergistically inhibit the proliferation, migration, invasion and adhesion of bone metastasis cells of prostate cancer, so that the bone metastasis effect of the prostate cancer is achieved.
In some embodiments, andrographolide and dioscin synergistically inhibit the Notch signaling pathway. In some embodiments, andrographolide and dioscin can inhibit the expression of key proteins of Notch signaling pathway and downstream genes (including Notch-1, NICD, hes-1) simultaneously. In some embodiments, andrographolide and dioscin can inhibit the expression of both key proteins of the Notch signaling pathway and downstream genes (including Notch-1, NICD, hes-1) in prostate cancer bone metastasis cells (e.g., PC-3).
In some embodiments, the molar concentration ratio of andrographolide to dioscin is (1.5-12): 1, and the combination of andrographolide and dioscin has a more excellent effect in inhibiting Notch signaling pathway activity than the combination of andrographolide and dioscin alone. In some embodiments, the molar concentration ratio of andrographolide to dioscin is (1.5-12) to 1, preferably (3-12): 1, more preferably (3-6): 1, 4.
In some embodiments, the dioscin includes a derivative form thereof. In some embodiments, the derivative form of dioscin includes a pharmaceutically acceptable salt of dioscin, a pharmaceutically acceptable ester of dioscin, a structural analog of dioscin, and a modification of dioscin. In some embodiments, the andrographolide comprises a derivative form thereof. In some embodiments, the andrographolide derivative forms include a pharmaceutically acceptable salt of andrographolide, a pharmaceutically acceptable ester of andrographolide, a structural analog of andrographolide, and a modification of andrographolide. In some embodiments, the modified product of dioscin refers to a product obtained by modifying dioscin, which does not change the substance of the product as a notch signaling pathway inhibitor. In some embodiments, the modifications of andrographolide refer to the products of andrographolide that have been modified without altering its substance as a notch signaling pathway inhibitor.
In some embodiments, andrographolide and dioscin are synergistic against bone metastasis from prostate cancer. In some embodiments, andrographolide and dioscin can synergistically achieve the following functions: (1) inhibiting the proliferation of prostate cancer bone metastases; (2) inhibiting migration of prostate cancer bone metastases; (3) inhibiting invasion of prostate cancer bone metastasis cells; and (4) inhibiting the adhesion of prostate cancer bone metastasis cells. In some embodiments, andrographolide and dioscin synergistically inhibit secretory expression of tumor metastasis associated proteins MMP-2 and MMP-9.
In some embodiments, the molar concentration ratio of andrographolide to dioscin is (1.5-12): 1, and the combined application of the andrographolide and the dioscin has more excellent effects in inhibiting the proliferation, migration, invasion, adhesion and tumor metastasis related protein expression of prostate cancer bone metastasis cells than the single application of either one of the andrographolide and the dioscin. In some embodiments, the molar concentration ratio of andrographolide to dioscin is (1.5-12) to 1, preferably (3-12): 1, more preferably (3-6): 1, 4.
In some embodiments, andrographolide and dioscin are capable of inhibiting the Notch signaling pathway in prostate cancer bone metastases cells.
In some embodiments, the dioscin includes a pharmaceutically acceptable salt or ester thereof. In some embodiments, the andrographolide comprises a pharmaceutically acceptable salt or ester thereof.
The beneficial effects of the invention are:
the invention unexpectedly finds that the andrographolide and dioscin are combined to play a synergistic role, more efficiently inhibits a notch signal channel, and further more efficiently plays a role in resisting the bone metastasis of the prostate cancer, including inhibiting the proliferation, migration, invasion, adhesion and secretory expression of proteins related to metastasis of bone metastasis cells of the prostate cancer.
Drawings
FIG. 1: cell proliferation after treating PC-3 cells with ANDRO of different concentrations;
FIG. 2 is a schematic diagram: cell proliferation after treating PC-3 cells with Dio at different concentrations;
FIG. 3: cell migration after 15 μ M ANDRO treatment of PC-3 cells;
FIG. 4: cell migration after 2.5. Mu.M, 5. Mu.M Dio treatment of PC-3 cells;
FIG. 5: cell invasion after treatment of PC-3 cells with 15. Mu.M ANDRO;
FIG. 6:2.5 μ M, 5 μ M Dio treated PC-3 cells after cell invasion;
FIG. 7 is a schematic view of: cell adhesion after treatment of PC-3 cells with 15. Mu.M ANDRO;
FIG. 8: cell adhesion after 2.5. Mu.M, 5. Mu.M Dio treatment of PC-3 cells;
FIG. 9: expression of Notch-1, hes-1 and NICD after 15. Mu.M ANDRO treatment of PC-3 cells, and GAPDH as an internal reference;
FIG. 10:2.5 μ M, 5 μ M Dio treated PC-3 cells after Notch-1, hes-1, NICD expression, GAPDH as internal reference;
in the above figures, an x indicates a significant difference compared to the control.
Detailed Description
The present invention will be more readily understood by the following examples, taken in conjunction with the general description of the invention, which are intended to illustrate embodiments of certain aspects of the invention, and are not intended to limit the invention. It should be noted that the experimental methods or experimental conditions used in the experimental examples were carried out according to the conventional methods or manufacturer's instructions without specific instructions, and the materials and reagents used in the experimental examples were commercially available without specific instructions.
Cell culture
Human prostate cancer bone metastasis PC-3 cell line was purchased from ATCC (American type culture Collection, USA), and the cells were seeded at 75cm 2 Culture flask 37 ℃ 5% 2 Medium culture, using 1640 medium supplemented with 10% fetal bovine serum, 100 units/ml penicillin and 100mg/ml streptomycin, taking logarithmic growth phase cells for testing.
MTS detection of PC-3 cell proliferation
PC-3 cells were digested with 0.25% trypsin and then used at a cell concentration of 3X 10 4 One cell/mL is evenly inoculated on a 96-well culture plate, each well is 100 mu L, drugs with different concentrations are added after the cells are attached to the wall, a DMSO control group and a blank control group (without cells) are set, each group is provided with 5 multiple wells, and the half inhibitory concentration (IC 50) of the PC-3 cells acted by the drugs is detected after 48 h. Followed by IC50PC-3 cells are treated by the drug, cultured for 0, 1,2 and 3d respectively, 10 mu L of MTS reagent is added into each well, incubated for 3h in a constant temperature box at 37 ℃, and the light absorption value A (490 nm) of each well is detected by a multifunctional microplate reader so as to observe the influence of the drug on cell proliferation along with the change of time.
Transwell detection of cell migration
PC-3 cells were treated in different groups for 48h, then resuspended by trypsinization, and the cell number was adjusted to 1X 10 6 one/mL, 200. Mu.L of cell suspension was inoculated into the upper chamber of a Transwell chamber, and 600. Mu.L of complete medium was added to the lower chamber, which was then placed in a 37 ℃ incubator for further 24h. After removing the Transwell chamber and carefully wiping the PC-3 cells on the membrane of the chamber, the PC-3 cells under the membrane were stained with 0.1% crystal violet, the number of cells in 5 high-power fields was randomly counted under a light microscope, and then the average value was calculated.
Transwell detection of cell invasion
Before the experiment, the mixture is mixed with 50mg/L Matrigel 1:8 dilution coated the upper chamber face of the bottom membrane of the Transwell chamber and air dried at 4 ℃. The subsequent steps were the same as the Transwell migration experiment described above.
Fibronectin detection of cell adhesion
After 2h plating with human fibronectin in 96-well plates, BSA incubation blocked specific binding sites for 30 min. PC-3 cells were treated in different groups for 48h, then resuspended by trypsinization, and the cell number was adjusted to 1X 10 5 And (2) inoculating 200uL of cell suspension into a 96-well plate, incubating for 30 minutes in an incubator at 37 ℃, washing the nonadherent cells by PBS, fixing for 30 minutes by paraformaldehyde, staining by 0.1% crystal violet, photographing and counting.
ELISA (enzyme-Linked immuno sorbent assay) for detecting secretory expression of MMP-2 and MMP-9
After treating the prostate cancer bone metastasis PC-3 cells for 24 hours by using medicaments with different concentrations, the determination is carried out by using human MMP-2 and MMP-9ELISA kits respectively. Adding 100 μ L of corresponding standard substance or cell supernatant into each well, sealing the ELISA plate at 37 deg.C, reacting for 90min, removing the liquid (not washing) in the ELISA plate, adding 100 μ L of biotin-labeled anti-human MMP-2 or MMP-9 antibody working solution into each well, and reacting at 37 deg.C for 60 min. After 3 times of PBS washing, 100. Mu.L of ABC working solution was added to each well and reacted at 37 ℃ for 30 minutes. After washing with PBS for 5 times, 100. Mu.L of TMB developing solution was added thereto and the reaction was carried out at 37 ℃ for 20 to 25 minutes in the absence of light. 100 μ L of TMB stop solution was added (at which time the blue color immediately turned yellow). And measuring absorbance by a microplate reader at 450nm, calculating corresponding concentration according to the standard curve, and dividing the concentration by the concentration of the control group to calculate the relative level.
Western blotting experiment (Western blot)
After digesting and resuspending PC-3 cells with trypsin, the cells were made 2X 10 cells with complete medium 5 each/mL of the cell suspension was uniformly inoculated into 6-well culture plates, and after each well was 2mL and 24h of cells adhered, the cells were treated with different concentrations of the drug. After 48h incubation, the cells were washed 3 times with pre-cooled PBS, then scraped with a cell scraper and transferred to a 15mL centrifuge tube, centrifuged at 4000r for ten minutes, the supernatant was discarded, 100 μ L of RIPA lysate and a protease inhibitor of PMSF (100) were added to each tube, transferred to a 1.ml ep tube, lysed on ice for 30min, then centrifuged at 12000r/min at 4 ℃ for 10min, the supernatant was aspirated, each set of total protein content was detected with a BCA kit and stained with a loading buffer, the total proteins were diluted to uniform concentration, denatured at 95 ℃ for 10min and stored at-20 ℃ for later use. 10% SDS-PAGE gels were dispensed with the concentrated gel and the quantified sample was added to the lane (approximately 50. Mu.g). And (4) carrying out 80V electrophoresis until the sample passes through a separation gel, then carrying out 120V electrophoresis, and then carrying out 300mA 90min electrotransfer. 5 BSA was subjected to 90min shaking table at room temperature, then washed 3 times, 10 min/time with TBST (p H = 7.6), followed by addition of Notch-1 antibody, NICD antibody, hes-1 antibody and internal reference GAPDH antibody, respectively, incubation overnight at 4 ℃,3 times, 10 min/time TBST after 1H shaking table the next time, followed by addition of Anti-mouse IgG (1: 2000) and Anti-rabbit IgG (1: 2000) labeled with horseradish peroxidase, incubation for 1H at room temperature, followed by washing 3 times, 10 min/time with TBST, and finally color exposure by ECL method.
Description of the invention: the relative ratio or ratio not clearly explained in the experimental results is a ratio converted in comparison with the non-administered control group, for example, if the non-administered group is 1 and the test result of the present group is 0.9, the relative ratio or ratio is 90%.
Results of cell proliferation experiments
Experimental group 1: different concentrations of ANDRO (2.5, 5, 10, 20, 40. Mu.M)
The results are shown in figure 1, and the inhibition degree of cell proliferation is more and more obvious and dose-dependent with the increase of the concentration of ANDRO. IC of cell proliferation inhibition Rate by SPSS statistical software analysis 50 About 15. Mu. Mol/L.
Experimental group 2: dio at different concentrations (2.5, 5, 10, 20, 40. Mu.M)
As shown in fig. 2, the inhibition of cell proliferation was more and more significant and dose-dependent with increasing concentration of ANDRO. IC of inhibition of cell proliferation by SPSS statistical software analysis 50 About 5. Mu. Mol/L.
Experimental group 3: mixing ANDRO and Dio at different concentration ratios
The ANDRO and Dio mixtures were configured according to the IC50, the groups with one of the drugs alone were tested simultaneously, and the amplitude of change was calculated relative to the equivalent drug alone in the manner: relative cell proliferation rate for the mixture application-relative cell proliferation rate for the single application.
Configuration schemes and test results are shown in the table below, and it can be seen that the combined use of ANDRO and Dio can synergistically inhibit cell proliferation with high efficiency.
| Combined medicine | |
|
Combination set 2 | Coupling group 3 | Coupling group 4 |
| Molar concentration ratio ANDRO：Dio | 12：1 | 6:1 | 3:1 | 1.5:1 | |
| ANDRO concentration/. |
15 | 15 | 15 | 15 | 15 |
| Dio concentration/. |
0 | 1.25 | 2.5 | 5 | 10 |
| Relative cell proliferation Rate | 61.74％ | 50.45％ | 28.52％ | 22.73％ | 20.99％ |
| Dio proliferation Rate Change relative to Dio alone | -48.12％ | -53.80％ | -42.25％ | -17.68％ |
Experimental results on migration and invasion abilities of cells
IC according to the drug 50 The effect of the concentration test drug on the migration and invasion ability of PC-3 cells was shown in FIGS. 3 and 4, and the migration ability of PC-3 cells was inhibited after the cells were treated with ANDRO (15. Mu.M) or DiO (2.5. Mu.M, 5. Mu.M). As a result, as shown in FIGS. 5 and 6, the invasion ability of PC-3 cells was inhibited after the cells were treated with ANDRO (15. Mu.M) or Dio (2.5. Mu.M, 5. Mu.M).
Designing a combined administration test according to the experimental results, configuring an ANDRO and Dio mixture, simultaneously testing a group using one of the drugs alone, and calculating the variation amplitude of the drug compared with the single use of the same amount of the drug, wherein the variation amplitude calculation mode comprises the following steps: mixture drug migration cell rate-drug alone migration cell rate, or mixture drug invasion cell rate-drug alone invasion cell rate.
Configuration protocols and test results as shown in the table below, it can be seen that combined administration of ANDRO and Dio can synergistically and efficiently inhibit cell migration and invasion.
| Combined medicine | |
Combination group 2 | Coupling group 3 |
| Molar ratio ANDRO: dio | 6:1 | 3:1 | |
| ANDROConcentration/. |
15 | 15 | 15 |
| Dio concentration/. |
0 | 2.5 | 5 |
| Rate of migration of cells | 32.88％ | 18.63％ | 12.49％ |
| Cell migration rate change with Dio alone | -44.70％ | -26.72％ | |
| Rate of invading cells | 48.65％ | 25.44％ | 22.71％ |
| Relative to Dio alone for cell invasion rate changes | -43.54％ | -23.83％ |
Results of cell adhesion experiments
IC according to the drug 50 The effect of the concentration-tested drugs on the adhesion ability of PC-3 cells, the results are shown in FIG. 7 andFIG. 8 shows that the cell adhesion between PC-3 cells was significantly reduced after ANDRO (15. Mu.M) or Dio (2.5. Mu.M, 5. Mu.M) treatment.
Designing a combined administration test according to the experimental results, configuring an ANDRO and Dio mixture, simultaneously testing a group using one of the drugs alone, and calculating the variation amplitude of the drug compared with the single use of the same amount of the drug, wherein the variation amplitude calculation mode comprises the following steps: mixture drug-adherent cell rate-the rate of adherent cells with drug alone.
Configuration protocol and test results are shown in the table below, and it can be seen that combined administration of ANDRO and Dio can synergistically inhibit cell adhesion ability with high efficiency.
| Combined medicine | |
Combination set 2 | Coupling group 3 |
| Molar ratio ANDRO: dio | 6:1 | 3:1 | |
| ANDRO concentration/. |
15 | 15 | 15 |
| Dio concentration/. |
0 | 2.5 | 5 |
| Rate of adherent cells | 49.84％ | 29.41％ | 30.52％ |
| Relative Dio adherent cell Rate Change alone | -21.63％ | -13.33％ |
Expression results of Notch-1, NICD, and Hes-1
According to the analysis, it is unexpectedly found that ANDRO and Dio can inhibit the activity of the Notch signaling pathway, specifically found that ANDRO (15 μ M) or Dio (2.5 μ M, 5 μ M) treated cells can inhibit the expression of key proteins and downstream genes of the Notch signaling pathway (including Notch-1, NICD, hes-1), and the results are shown in FIG. 9 and FIG. 10.
The effect of the combined administration was further tested and the results are shown in the following table, it can be seen that the combined use of ANDRO and Dio has a superior effect of inhibiting Notch signaling pathway activity.
| Medicament | |
Single use group 2 | Single use group 3 | Combination group 2 | Coupling group 3 |
| Molar ratio ANDRO: dio | 6:1 | 3:1 | |||
| ANDRO concentration/. |
15 | 0 | 0 | 15 | 15 |
| Dio concentration/. |
0 | 2.5 | 5 | 2.5 | 5 |
| Relative expression Rate of Notch-1 | 84.85％ | 52.35％ | 34.67％ | 25.64％ | 20.53％ |
| Relative expression Rate of Hes-1 | 75.36％ | 90.53％ | 80.55％ | 60.44％ | 53.52％ |
| Relative NICD expression Rate | 40.24％ | 74.24％ | 56.27％ | 23.65％ | 19.69％ |
Test results of secretory expression of tumor metastasis related proteins MMP2 and MMP9
According to the results of the above experiments, the secretion expression levels of the tumor metastasis associated proteins MMP2 and MMP9 in PC-3 cells by the single administration and the combined administration were simultaneously measured, and the results are shown in relative secretion rates, as shown in the following table, it can be seen that the expression of the tumor metastasis associated proteins MMP-2 and MMP-9 can be inhibited both by using the ANDRO or Dio alone and by using the ANDRO and Dio in combination, and the inhibition efficiency of the combined administration is better.
| Medicament | |
Single use group 2 | Single use group 3 | Combination group 2 | Coupling group 3 |
| Molar ratio ANDRO: dio | 6:1 | 3:1 | |||
| ANDRO concentration/. |
15 | 0 | 0 | 15 | 15 |
| Dio concentration/. |
0 | 2.5 | 5 | 2.5 | 5 |
| Relative MMP-2 secretion rate | 43.62％ | 65.37％ | 60.03％ | 22.56％ | 24.34％ |
| Relative MMP-9 secretion rate | 37.99％ | 72.55％ | 61.34％ | 19.86％ | 20.85％ |
The above description is only exemplary of the present invention, and the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily made by those skilled in the art within the technical scope of the present invention disclosed herein should be covered within the scope of the present invention.
Claims (4)
1. The pharmaceutical composition for resisting the bone metastasis of the prostate cancer is characterized in that the active ingredients of the pharmaceutical composition are andrographolide and dioscin, wherein the molar concentration ratio of the andrographolide to the dioscin is (1.5-12) to 1.
2. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition for resisting prostate cancer bone metastasis has at least one of the following functions: (1) inhibiting the proliferation of prostate cancer bone metastases; (2) inhibiting migration of prostate cancer bone metastases; (3) inhibiting invasion of prostate cancer bone metastasis cells; and (4) inhibiting the adhesion of prostate cancer bone metastasis cells.
3. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition for resisting prostate cancer bone metastasis has a function of inhibiting Notch signaling pathway.
4. The pharmaceutical composition according to claim 1, wherein the dioscin comprises a pharmaceutically acceptable salt thereof; and/or, the andrographolide comprises a pharmaceutically acceptable salt thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010219733.XA CN111317740B (en) | 2020-03-25 | 2020-03-25 | Notch signal pathway inhibition composition and application of composition in resisting prostatic cancer bone metastasis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010219733.XA CN111317740B (en) | 2020-03-25 | 2020-03-25 | Notch signal pathway inhibition composition and application of composition in resisting prostatic cancer bone metastasis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111317740A CN111317740A (en) | 2020-06-23 |
| CN111317740B true CN111317740B (en) | 2022-12-16 |
Family
ID=71165945
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010219733.XA Active CN111317740B (en) | 2020-03-25 | 2020-03-25 | Notch signal pathway inhibition composition and application of composition in resisting prostatic cancer bone metastasis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111317740B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105663858A (en) * | 2016-03-16 | 2016-06-15 | 张淑振 | Traditional Chinese medicine for treating dental ulcer |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8142819B2 (en) * | 2002-10-21 | 2012-03-27 | Metaproteomics, Llc | Synergistic compositions that treat or inhibit pathological conditions associated with inflammatory response |
-
2020
- 2020-03-25 CN CN202010219733.XA patent/CN111317740B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105663858A (en) * | 2016-03-16 | 2016-06-15 | 张淑振 | Traditional Chinese medicine for treating dental ulcer |
Non-Patent Citations (2)
| Title |
|---|
| 穿心莲内酯对前列腺癌PC-3细胞的凋亡作用及机制研究;郭明伟等;《中药药理与临床》;20151215;第31卷(第6期);摘要 * |
| 薯蓣皂苷的现代药理学研究进展;陈亚琴等;《现代中西医结合杂志》;20190810;第28卷(第23期);第2613页右栏第1段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111317740A (en) | 2020-06-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | Scoparone improves hepatic inflammation and autophagy in mice with nonalcoholic steatohepatitis by regulating the ROS/P38/Nrf2 axis and PI3K/AKT/mTOR pathway in macrophages | |
| EP3280421B1 (en) | Pharmaceutical compositions for combination therapy | |
| EP1048673A1 (en) | Neovascularization inhibitor containing dienogest as the active ingredient | |
| Wang et al. | Dictamnine promotes apoptosis and inhibits epithelial-mesenchymal transition, migration, invasion and proliferation by downregulating the HIF-1α and Slug signaling pathways | |
| US6890946B2 (en) | Use of parthenolide to inhibit cancer | |
| Lin et al. | Berberrubine attenuates potassium oxonate-and hypoxanthine-induced hyperuricemia by regulating urate transporters and JAK2/STAT3 signaling pathway | |
| Shen et al. | Anti-proliferation and anti-inflammation effects of corilagin in rheumatoid arthritis by downregulating NF-κB and MAPK signaling pathways | |
| Wang et al. | Blockade of TNF-α-induced NF-κB signaling pathway and anti-cancer therapeutic response of dihydrotanshinone I | |
| Yang et al. | Gypenoside XLIX protects against acute kidney injury by suppressing IGFBP7/IGF1R-mediated programmed cell death and inflammation | |
| Guo et al. | S-petasin induces apoptosis and inhibits cell migration through activation of p53 pathway signaling in melanoma B16F10 cells and A375 cells | |
| Niu et al. | ASIC1a promotes synovial invasion of rheumatoid arthritis via Ca2+/Rac1 pathway | |
| RU2750992C2 (en) | Pharmaceutical composition for use in the treatment of prostate pathologies | |
| Zhuang et al. | MCTR3 reduces LPS-induced acute lung injury in mice via the ALX/PINK1 signaling pathway | |
| Wang et al. | 6-Shogaol reduces progression of experimental endometriosis in vivo and in vitro via regulation of VGEF and inhibition of COX-2 and PGE2-mediated inflammatory responses | |
| CN111317740B (en) | Notch signal pathway inhibition composition and application of composition in resisting prostatic cancer bone metastasis | |
| Yang et al. | Green tea catechins attenuate human primary pterygium cell survival and migration via modulation of ERK p42/p44 and p38 pathways | |
| Lu et al. | Isoalantolactone inhibits RANKL-induced osteoclast formation via multiple signaling pathways | |
| US20240050444A1 (en) | Treatment methods | |
| Holmström et al. | Current perspectives on the treatment of nasal polyposis: a Swedish opinion report | |
| Li et al. | Bioactive peptides sensitize cells to anticancer effects of oxaliplatin in human colorectal cancer xenografts in nude mice | |
| CN111658655A (en) | Application of cucurbitacin B in preparation of iron death inducer and anti-nasopharyngeal carcinoma drug | |
| Liu et al. | Z-Guggulsterone alleviates renal fibrosis by mitigating G2/M cycle arrest through Klotho/p53 signaling | |
| US20160120857A1 (en) | Anti-Fibrogenic Compounds, Methods and Uses Thereof | |
| KR20170036928A (en) | IKK Pharmaceutical Composition for Preventing or Treating Chronic Rhinosinusitis Comprising IKK Inhibitor | |
| KR20220094614A (en) | A composition comprising ginseng for preventing, improving or treating chronic obstructive pulmonary disease |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |