CN111166887B - Pharmaceutical composition of baicalein and autophagy inhibitor and application thereof - Google Patents
Pharmaceutical composition of baicalein and autophagy inhibitor and application thereof Download PDFInfo
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Abstract
The invention relates to a pharmaceutical composition of baicalein and an autophagy inhibitor and application thereof, in particular to application of the combination of the baicalein and the autophagy inhibitor in treating tumors. The inventor finds a series of autophagy inhibitors combined with baicalein, finds that the combination of the two inhibitors generates a synergistic effect, can effectively improve the anti-tumor cell proliferation capacity of the medicament, provides a foundation for clinical combined administration of the baicalein and the autophagy inhibitors, improves the tumor treatment effect, can reduce the medicament cost, and has important clinical application value.
Description
Technical Field
The application belongs to the field of medicines, and particularly relates to a pharmaceutical composition of baicalein and an autophagy inhibitor and application thereof.
Background
The baicalein is one of the main components of natural Chinese herbal medicine scutellaria baicalensis, is also an important flavonoid compound in the scutellaria baicalensis, is widely concerned by scholars at home and abroad due to multiple pharmacological actions, has the effects of resisting tumors, protecting heart and cerebral vessels, protecting nerves, treating or preventing diabetes and complications thereof, removing oxygen radicals, resisting oxidation, resisting bacteria and viruses and the like, and has good commercial development value.
Autophagy belongs to programmed cell death, and when an organism faces exogenous stimulation which is unfavorable for growth, such as hunger, oxidative stress, infection and the like, in order to adapt to a severe external environment, autophagy is started, and non-functional and damaged organelles are digested and recycled to be degraded into reusable small molecular proteins, polypeptides, amino acids and the like, so that the organelles participate in substance energy circulation, the purposes of waste utilization and self-purification are achieved, the cell viability is enhanced, and the autophagy plays an important role in cell metabolism. The effects of autophagy on tumor cells are complex and exhibit diametrically opposite effects of either inhibition or promotion. As to what role autophagy plays in the development of tumor, the specific situation should be analyzed specifically, and the research shows that the effect of the antitumor drug is related to the antitumor mechanism of the drug in addition to the intracellular autophagy level, so that how to cause more autophagic cell death while eliminating the protection of autophagy on cells is a considerable direction for further research.
Based on the thought of the combined action of Chinese and western medicines, a series of autophagy inhibitors (MRT 68921, 3-MA and chloroquine) combined with baicalein and a synergistic interval in combination are found, and the baicalein and the autophagy inhibitors provide a basis for clinical combined medication.
Disclosure of Invention
The invention aims to provide a pharmaceutical composition for treating tumors, which consists of baicalein and an autophagy inhibitor.
Autophagy refers to the phenomenon of autophagy, including macroautophagy, chaperone-mediated autophagy and microautophagy, which are 3 main modes, wherein the process of degrading long-life proteins and damaged organelles in cells through a lysosome pathway is a phenomenon peculiar to eukaryotic cells, and the macroautophagy is generally called autophagy. The autophagy signal transduction pathways mainly include mTOR-dependent pathways and mTOR-independent pathways.
Autophagy inhibitors include inhibitors of phosphatidylinositol 3-kinase (PI3K), chloroquine, Thapsigargin (TG), baverromycin A1, (+ -) -Bay K8644, Desipramine (DCMI), Trichostatin (TSA), vorinostat (SAHA), phenylacetylene sulfonamide (PES), and the like.
The inhibitor of phosphatidylinositol 3-kinase (PI3K) specifically further includes: 3-methyladenosine (3-MA), Wortmannin (Wortmannin), LY294002 and the like.
Further, the autophagy inhibitor includes an ULK1/2 inhibitor, a PI3K inhibitor, chloroquine, or the like.
Preferably, the autophagy inhibiting agent includes MRT68921, 3-MA, chloroquine, and the like.
MRT68921 is a highly potent dual autophagy kinase ULK1/2 inhibitor. ULK1 is a serine/threonine protein kinase important for the initiation of autophagy, and therefore MRT68921 inhibits the early stages of autophagy.
3-methyladenine (3-MA) is an inhibitor of phosphatidylinositol 3-kinase (PI 3K). PI3K plays an important role in many biological processes, including controlling the activation of mTOR, a key regulator of autophagy. 3-MA inhibits autophagy by inhibiting class III PI3K from blocking autophagosome formation, and is also an early inhibitor of autophagy.
Chloroquine (CQ) can increase the pH of lysosomes, thereby inhibiting the fusion of autophagosomes with lysosomes and the degradation of lysosomal proteins, and the change of the pH value in the lysosome by Chloroquine leads to the reduction of the stability of the lysosome membrane, further leads to the depolarization of the mitochondrial membrane and the reduction of the membrane potential, and the reduction of the mitochondrial membrane potential activates Caspase3 and PARP to induce apoptosis through a Caspase-dependent pathway. Autophagy is inhibited at the later stage of autophagy.
Furthermore, the pharmaceutical composition also comprises pharmaceutic adjuvants.
The pharmaceutical excipients are selected from one or more of diluents, flavoring agents, lubricants, glidants, and optionally disintegrants and binders.
The diluent is selected from one or more of pregelatinized starch, mannitol, calcium hydrogen phosphate, microcrystalline cellulose 101 and xylitol;
the flavoring agent is selected from one or more of mint-flavored powdered essence, blackcurrant-flavored powdered essence, strawberry-flavored powdered essence and cream-flavored powdered essence;
the lubricant is selected from one or more of magnesium stearate, superfine silica gel powder, talcum powder and hydrogenated vegetable oil;
the glidant is selected from silicon dioxide;
the disintegrating agent is selected from one or more of dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl methylcellulose and cross-linked polyvinylpyrrolidone;
the adhesive is selected from one or more of starch slurry, polyvidone, gelatin and polyethylene glycol.
Further, the concentration ratio of baicalein to ULK1/2 inhibitor is 1: 0.0125-0.16.
Preferably, the concentration ratio of baicalein to MRT68921 is 1: 0.0125-0.16.
More preferably, the concentration ratio of baicalein to MRT68921 is 1: 0.0125-0.1 or 1: 0.02-0.16.
Further, the concentration ratio of baicalein to the PI3K inhibitor is 1: 15.6-200.
Preferably, the concentration ratio of the baicalein to the 3-MA is 1: 15.6-200.
More preferably, the concentration ratio of baicalein to 3-MA is 1: 15.6-125 or 1: 25-200.
Further, the concentration ratio of the baicalein to the chloroquine inhibitor is 1: 0.063-0.8.
More preferably, the concentration ratio of baicalein to chloroquine is 1: 0.1-0.4 or 1:0.063-0.5 or 1: 0.1-0.8.
The invention aims to provide application of the pharmaceutical composition in preparing a medicine for treating tumors.
Tumors include, but are not limited to: fibrosarcoma, myosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, thyroid cancer, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchial carcinoma, hepatoma, liver cancer, bile duct cancer, choriocarcinoma, seminoma, embryonal carcinoma, wilms' tumor, cervical cancer, testicular tumor, lung cancer, small cell lung cancer, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, papillary carcinoma, thyroid carcinoma, and other tumors, Glioblastoma, retinoblastoma, leukemia, polycythemia vera, lymphoma, multiple myeloma.
Preferably, the tumor is lung cancer, thyroid cancer or glioblastoma.
Further, the application of the pharmaceutical composition in preparing anti-tumor cell proliferation medicines.
Further, the concentration ratio of baicalein to ULK1/2 inhibitor is 1: 0.0125-0.16.
Preferably, the concentration ratio of baicalein to MRT68921 is 1: 0.0125-0.16.
More preferably, for treating thyroid cancer, the concentration ratio of baicalein to MRT68921 is 1: 0.0125-0.1, the concentration ratio of baicalein to MRT68921 is 1: 0.02-0.16.
Further, the concentration ratio of baicalein to the PI3K inhibitor is 1: 15.6-200.
Preferably, the concentration ratio of the baicalein to the 3-MA is 1: 15.6-200.
More preferably, in the treatment of thyroid cancer, the concentration ratio of baicalein to 3-MA is 1: 15.6-125, the concentration ratio of baicalein to 3-MA is 1: 25-200.
Further, the concentration ratio of the baicalein to the chloroquine inhibitor is 1: 0.063-0.8.
More preferably, in the treatment of lung cancer, the concentration ratio of baicalein to chloroquine is 1: 0.1-0.4, the concentration ratio of baicalein to chloroquine is 1:0.063-0.5 when treating thyroid cancer, and the concentration ratio of baicalein to chloroquine is 1: 0.1-0.8.
The dosage form of the pharmaceutical composition of the present invention is not particularly limited, and the pharmaceutical composition may be prepared into one or more of tablets, capsules, pills, mixtures, gels, ointments, granules, powders, ointments, granules, pellets, oral liquids, dripping pills, injections or injections, or any other dosage forms.
The invention has the advantages that:
the combined medication of the baicalein and the autophagy inhibitor can obviously improve the anti-tumor cell proliferation capacity of the medicament, generates a synergistic effect, can effectively reduce the medicament cost, improves the clinical treatment effect, and also provides a theoretical basis for the clinical combined medication of Chinese and western medicaments.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are intended to be illustrative only and are not to be construed as limiting the invention. Those of ordinary skill in the art will understand that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents. The following examples are examples of experimental methods not indicating specific conditions, and the detection is usually carried out according to conventional conditions or according to the conditions recommended by the manufacturers.
Example 1 Combined administration experiment of baicalein and MRT68921 in Lung cancer cells
1.1 Experimental methods
Cell culture:
digesting non-small cell lung cancer H1299 cells to prepare a cell suspension, inoculating 5000 cells per well into a 96-well plate, placing at 37 ℃ and 5% CO2The culture was carried out overnight in an incubator. On the next day, the cells are divided into groups, the corresponding culture medium is added into a control group, then other groups are added with different drugs for treatment, and the culture is continued for 24 hours, wherein in the combined administration group, the baicalein is added after the autophagy inhibitor MRT68921 is pretreated for 1 hour.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administered concentrations of MRT68921 were: control group 0. mu.M, group 5 1. mu.M, group 6 2. mu.M, group 7 4. mu.M, group 8. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M MRT68921, group 9 50 μ M baicalein +0 μ M MRT68921, group 10 50 μ M baicalein +1 μ M MRT68921, group 11 50 μ M baicalein +2 μ M MRT68921, group 12 50 μ M baicalein +4 μ M MRT68921, and group 13 50 μ M baicalein +8 μ M MRT 68921.
Determination of IC50 value and determination of efficacy of combination:
(1) calculation of median inhibitory concentration (IC50) values: half maximal inhibitory concentration (IC50) values were calculated using SPSS 23.0 software based on the effect of different concentrations of drug on the cells.
(2) After 24 hours of administration, the activity of each group of cells was measured by MTT method and the inhibition rate was calculated (inhibition rate ═ control OD value-administered group OD value)/(control OD value-blank group OD value) × 100%).
(3) And (3) judging the effect of the double-medicine combination:
the calculation method for judging the effect of the combined medication comprises the following steps: and judging by adopting a gold positive mean q value method. The q value is obtained by the following formula: q ═ PA+B/(PA+PB-PA×PB). In the formula PA、PBAnd PA+BRespectively the treatment rates of the A medicine group, the B medicine group and the combination of the two medicines. q is less than 1, which indicates that the two medicines generate antagonism after being used together; q is more than 1, which indicates that the two drugs produce synergistic effect after being combined together, and q is 1, which indicates that the two drugs produce additive effect after being combined together.
1.2 results of the experiment
The results show that both baicalein and MRT68921 have the effect of inhibiting the growth of tumor cells on H1299 cells. Calculating the half inhibition concentration (IC50) value by using SPSS 23.0 software, wherein the average inhibition rates of baicalein at 25, 50, 100 and 200 mu M are respectively 16.6 +/-1.0%, 30.1 +/-4.4%, 72.0 +/-3.9% and 86.0 +/-4.9%, and the half inhibition concentration (IC50) value is calculated to be 68.4 +/-6.4; similarly, the IC50 value for MRT68921 was calculated to be 11.3. + -. 0.9. In combination, baicalein and MRT68921 are administered in a ratio of 1: has synergistic effect at 0.02-0.16.
TABLE 1 combination of baicalein and MRT68921 for administration in lung cancer cells
Example 2 Combined administration of baicalein and MRT68921 experiments in thyroid cancer cells
2.1 Experimental methods
Cell culture: digesting thyroid cancer TPC-1 cells to prepare cell suspension, inoculating 5000 cells per well into 96-well plate, placing at 37 deg.C and 5% CO2The culture was carried out overnight in an incubator. On the next day, the cells are divided into groups, the corresponding culture medium is added into a control group, then other groups are added with different drugs for treatment, and the culture is continued for 24 hours, wherein in the combined administration group, the baicalein is added after the autophagy inhibitor MRT68921 is pretreated for 1 hour.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administered concentrations of MRT68921 were: control group 0. mu.M, group 5 1. mu.M, group 6 2. mu.M, group 7 4. mu.M, group 8. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M MRT68921, group 9 80 μ M baicalein +0 μ M MRT68921, group 10 80 μ M baicalein +1 μ M MRT68921, group 11 80 μ M baicalein +2 μ M MRT68921, group 12 80 μ M baicalein +4 μ M MRT68921, and group 13 80 μ M baicalein +8 μ M MRT 68921.
2.2 results of the experiment
The results show that both baicalein and MRT68921 have the effect of inhibiting the growth of tumor cells on thyroid cancer cell TPC-1. Calculating the half inhibition concentration (IC50) value by using SPSS 23.0 software, wherein the average inhibition rates of baicalein at 25, 50, 100 and 200 mu M are respectively 7.8 +/-2.1%, 18.0 +/-0.8%, 61.0 +/-0.9% and 80.1 +/-1.9%, and the half inhibition concentration (IC50) value is 91.6 +/-1.4; also, the IC50 value for MRT68921 was calculated to be 11.8. + -. 1.6. When the combination is administrated, each group shows synergistic effect, wherein the best effect is achieved when the ratio of baicalein to MRT68921 is 1: 0.05.
TABLE 2 combination of baicalein and MRT68921 for administration in thyroid cancer cells
Example 3 Combined baicalein and MRT68921 dosing experiments in glioblastoma cells
3.1 Experimental methods
Cell culture: glioblastoma U87-MG cells were digested to prepare a cell suspension, 5000 cells per well were inoculated into a 96-well plate, and the plate was incubated at 37 ℃ with 5% CO2The culture was carried out overnight in an incubator. On the next day, the cells are divided into groups, the corresponding culture medium is added into a control group, then other groups are added with different drugs for treatment, and the culture is continued for 24 hours, wherein in the combined administration group, the baicalein is added after the autophagy inhibitor MRT68921 is pretreated for 1 hour.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administered concentrations of MRT68921 were: control group 0. mu.M, group 5 1. mu.M, group 6 2. mu.M, group 7 4. mu.M, group 8. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M MRT68921, group 9 50 μ M baicalein +0 μ M MRT68921, group 10 50 μ M baicalein +1 μ M MRT68921, group 11 50 μ M baicalein +2 μ M MRT68921, group 12 50 μ M baicalein +4 μ M MRT68921, and group 13 50 μ M baicalein +8 μ M MRT 68921.
3.2 results of the experiment
The results show that both baicalein and MRT68921 have the effect of inhibiting the growth of tumor cells on human brain astrocytoma cell U87-MG, and the IC50 value of baicalein is 150 +/-7.9 and the IC50 value of MRT68921 is 7.5 +/-0.8 through experiments and calculation. When the combination was administered, each group showed a synergistic effect, however, the synergistic effect tended to decrease with increasing ratio of MRT 68921.
TABLE 3 combination of baicalein and MRT68921 in glioblastoma cells
| Inhibition ratio (%) | Q value | Ratio of | |
| Group 9 | 32.2±5.1％ | — | — |
| Group 10 | 44.3±5.6％ | 1.20 | 1：0.02 |
| Group 11 | 51.6±3.3％ | 1.14 | 1：0.04 |
| Group 12 | 60.2±2.0％ | 1.16 | 1：0.08 |
| Group 13 | 75.7±2.5％ | 1.11 | 1：0.16 |
Example 4 Combined administration experiment of baicalein and 3-MA in Lung cancer cells
4.1 Experimental methods
Cell culture: reference is made to example 1.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administration concentrations of 3-MA were: control group 0. mu.M, group 5 1.25. mu.M, group 6 2.5. mu.M, group 7 5. mu.M, group 8 10. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M3-MA, group 9 50 μ M baicalein +0 μ M3-MA, group 10 50 μ M baicalein +1.25 μ M3-MA, group 11 50 μ M baicalein +2.5 μ M3-MA, group 12 50 μ M baicalein +5 μ M3-MA, and group 13 50 μ M baicalein +10 μ M3-MA.
4.2 results of the experiment
The results show that both baicalein and 3-MA have the effect of inhibiting the growth of the lung cancer cells H1299. Calculating the half inhibition concentration (IC50) value by using SPSS 23.0 software, wherein the average inhibition rates of baicalein at 25, 50, 100 and 200 mu M are respectively 16.6 +/-1.0%, 30.1 +/-4.4%, 72.0 +/-3.9% and 86.0 +/-4.9%, and the half inhibition concentration (IC50) value is calculated to be 68.4 +/-6.4; similarly, the IC50 value for 3-MA was calculated to be 16.7. + -. 2.2. When the combination is administered, each group shows synergistic effect, and the synergistic effect shows increasing trend along with the increase of the 3-MA ratio.
TABLE 4 Combined administration of baicalein and 3-MA in Lung cancer cells
| Inhibition ratio (%) | Q value | Ratio of | |
| Group 9 | 36.2±4.1％ | — | — |
| Group 10 | 41.0±2.1％ | 1.13 | 1：25 |
| Group 11 | 52.2±3.8％ | 1.28 | 1：50 |
| Group 12 | 58.3±4.0％ | 1.34 | 1：100 |
| Group 13 | 71.0±0.9％ | 1.34 | 1：200 |
Example 5 Combined administration of baicalein and 3-MA in thyroid cancer cells
5.1 Experimental methods
Cell culture: reference is made to example 2.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administration concentrations of 3-MA were: control group 0. mu.M, group 5 1.25. mu.M, group 6 2.5. mu.M, group 7 5. mu.M, group 8 10. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M3-MA, group 9 80 μ M baicalein +0 μ M3-MA, group 10 80 μ M baicalein +1.25 μ M3-MA, group 11 80 μ M baicalein +2.5 μ M3-MA, group 12 80 μ M baicalein +5 μ M3-MA, and group 13 80 μ M baicalein +10 μ M3-MA.
5.2 results of the experiment
The results show that both baicalein and 3-MA have the effect of inhibiting the growth of the tumor cells on the thyroid cancer cells TPC-1. The half inhibitory concentration (IC50) values were calculated using SPSS 23.0 software, the mean inhibition ratios were 7.8. + -. 2.1%, 18.0. + -. 0.8%, 61.0. + -. 0.9%, 80.1. + -. 1.9% for baicalein at 25, 50, 100, 200. mu.M, the half inhibitory concentration (IC50) value was 91.6. + -. 1.4 using SPSS 23.0 software, and similarly, the IC50 value for 3-MA was 13.1. + -. 1.0. When the combination is administrated, each group shows synergistic effect, and the synergistic effect is in a descending trend along with the increase of the 3-MA ratio.
TABLE 5 combination of baicalein and 3-MA in thyroid cancer cells
| Inhibition ratio (%) | Q value | Ratio of | |
| Group 9 | 41.9±1.3％ | — | — |
| Group 10 | 78.9±3.1％ | 1.84 | 1：15.6 |
| Group 11 | 81.5±1.8％ | 1.69 | 1：31.3 |
| Group 12 | 82.8±1.7％ | 1.55 | 1：62.5 |
| Group 13 | 85.1±3.8％ | 1.29 | 1：125 |
Example 6 Combined baicalein and 3-MA administration experiments in glioblastoma cells
6.1 Experimental methods
Cell culture: reference is made to example 3.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administration concentrations of 3-MA were: control group 0. mu.M, group 5 1.25. mu.M, group 6 2.5. mu.M, group 7 5. mu.M, group 8 10. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M3-MA, group 9 50 μ M baicalein +0 μ M3-MA, group 10 50 μ M baicalein +1.25 μ M3-MA, group 11 50 μ M baicalein +2.5 μ M3-MA, group 12 50 μ M baicalein +5 μ M3-MA, and group 13 50 μ M baicalein +10 μ M3-MA.
6.2 results of the experiment
The results show that both baicalein and 3-MA have the effect of inhibiting the growth of tumor cells on human brain astrocytoma cells U87-MG. Through experiments and calculation, the IC50 value of baicalein is 150.0 +/-7.9, and the IC50 value of 3-MA is 13.9 +/-2.3. When administered in combination, each group showed synergistic effects.
TABLE 6 combination of baicalein and 3-MA in glioblastoma cells
| Inhibition ratio (%) | Q value | Ratio of | |
| Group 9 | 30.3±5.0％ | — | — |
| Group 10 | 45.8±4.3％ | 1.29 | 1：25 |
| Group 11 | 49.1±5.7％ | 1.15 | 1：50 |
| Group 12 | 58.8±4.0％ | 1.22 | 1：100 |
| Group 13 | 73.8±3.5％ | 1.23 | 1：200 |
Example 7 Combined administration experiment of baicalein and chloroquine in Lung cancer cells
7.1 Experimental methods
Cell culture: reference is made to example 1.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administration concentration of chloroquine is as follows: control group 0. mu.M, group 5. mu.M, group 6 10. mu.M, group 7 20. mu.M, group 8 40. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M chloroquine, group 9 50 μ M baicalein +0 μ M chloroquine, group 10 50 μ M baicalein +5 μ M chloroquine, group 11 50 μ M baicalein +10 μ M chloroquine, group 12 50 μ M baicalein +20 μ M chloroquine, and group 13 50 μ M baicalein +40 μ M chloroquine.
7.2 results of the experiment
The result shows that the baicalein has the function of inhibiting the growth of the tumor cells on lung cancer cell tumor cells H1299. The half maximal inhibitory concentration (IC50) value was calculated to be 68.4 ± 6.4 using SPSS 23.0 software; the average inhibition rates of baicalein at 25, 50, 100 and 200 mu M are respectively 16.6 +/-1.0%, 30.1 +/-4.4%, 72.0 +/-3.9% and 86.0 +/-4.9%, and the half inhibition concentration (IC50) value calculated by SPSS 23.0 software is 68.4 +/-6.4, so that the cytotoxicity of CQ is relatively small, and the IC50 cannot be calculated. When the combination is administrated, the ratio of the baicalein to the chloroquine is 1: 0.1-0.4 has synergistic effect.
TABLE 7 Combined administration of baicalein and chloroquine in Lung cancer cells
| Inhibition ratio (%) | Q value | Ratio of | |
| Group 9 | 36.9±4.2％ | — | — |
| Group 10 | 49.4±1.4％ | 1.34 | 1：0.1 |
| Group 11 | 47.7±3.4％ | 1.21 | 1：0.2 |
| Group 12 | 48.2±5.2％ | 1.17 | 1：0.4 |
| Group 13 | 40.7±3.9％ | 0.93 | 1：0.8 |
Example 8 combination of baicalein and chloroquine in thyroid cancer cells
8.1 Experimental methods
Cell culture: reference is made to example 2.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administration concentration of chloroquine is as follows: control group 0. mu.M, group 5. mu.M, group 6 10. mu.M, group 7 20. mu.M, group 8 40. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M chloroquine, group 9 80 μ M baicalein +0 μ M chloroquine, group 10 80 μ M baicalein +5 μ M chloroquine, group 11 80 μ M baicalein +10 μ M chloroquine, group 12 80 μ M baicalein +20 μ M chloroquine, and group 13 80 μ M baicalein +40 μ M chloroquine.
8.2 results of the experiment
The result shows that the baicalein has the function of inhibiting the growth of the tumor cells on the thyroid cancer cells TPC-1. The half inhibition concentration (IC50) value is calculated by adopting SPSS 23.0 software, the average inhibition rates of baicalein at 25, 50, 100 and 200 mu M are respectively 7.8 +/-2.1%, 18.0 +/-0.8%, 61.0 +/-0.9% and 80.1 +/-1.9%, the half inhibition concentration (IC50) value is calculated to be 91.6 +/-1.4, the cytotoxicity of CQ is relatively small, and the IC50 cannot be calculated. When the combination is administrated, the ratio of the baicalein to the chloroquine is 1:0.063-0.5 has synergistic effect, and the synergistic effect is decreased with the increase of chloroquine proportion.
TABLE 8 combination of baicalein and chloroquine in thyroid carcinoma cells
| Inhibition ratio (%) | Q value | Ratio of | |
| Group 9 | 46.5±3.6％ | — | — |
| Group 10 | 82.1±2.5％ | 1.73 | 1：0.063 |
| Group 11 | 82.5±2.1％ | 1.68 | 1：0.125 |
| Group 12 | 75.1±1.3％ | 1.48 | 1：0.25 |
| Group 13 | 57.0±2.5％ | 1.01 | 1：0.5 |
Example 9 Combined baicalein and chloroquine administration experiments in glioblastoma cell U87-MG cells
9.1 Experimental methods
Cell culture: reference is made to example 3.
Grouping experiments:
the administration concentration of the baicalein is as follows: control group 0. mu.M, group 1 25. mu.M, group 2 50. mu.M, group 3 100. mu.M, group 4 200. mu.M;
the administration concentration of chloroquine is as follows: control group 0. mu.M, group 5. mu.M, group 6 10. mu.M, group 7 20. mu.M, group 8 40. mu.M;
the concentrations administered for the combined administration were: control group 0 μ M baicalein +0 μ M chloroquine, group 9 50 μ M baicalein +0 μ M chloroquine, group 10 50 μ M baicalein +5 μ M chloroquine, group 11 50 μ M baicalein +10 μ M chloroquine, group 12 50 μ M baicalein +20 μ M chloroquine, and group 13 50 μ M baicalein +40 μ M chloroquine.
9.2 results of the experiment
The result shows that the baicalein has the function of inhibiting the growth of tumor cells on human brain astrocytoma cells U87-MG. SPSS 23.0 software is adopted to calculate the median inhibitory concentration (IC50) value, baicalein is 150.0 +/-7.9, the cytotoxic effect of CQ is small, and IC50 cannot be calculated. When the combination is administrated, the ratio of the baicalein to the chloroquine is 1: 0.1-0.8, the synergistic effect is decreased with the increase of CQ ratio.
TABLE 9 Combined administration of baicalein and chloroquine in glioblastoma cells
Claims (5)
1. The application of a pharmaceutical composition in preparing a medicament for treating lung cancer, the pharmaceutical composition comprises baicalein and MRT68921, and is characterized in that the concentration ratio of the baicalein to the MRT68921 is 1: 0.02-0.16.
2. The application of a pharmaceutical composition in preparing a medicament for treating thyroid cancer is disclosed, wherein the pharmaceutical composition comprises baicalein and MRT68921, and is characterized in that the concentration ratio of the baicalein to the MRT68921 is 1: 0.0125-0.1.
3. The application of a pharmaceutical composition in preparing a medicament for treating glioblastoma, the pharmaceutical composition consists of baicalein and MRT68921, and is characterized in that the concentration ratio of the baicalein to the MRT68921 is 1: 0.02-0.16.
4. Use according to any one of claims 1 to 3, wherein the pharmaceutical composition is for the manufacture of a medicament for the treatment of tumor cell proliferation.
5. Use according to any one of claims 1 to 3, wherein the pharmaceutical composition further comprises a pharmaceutical excipient selected from one or more of diluents, flavoring agents, lubricants, glidants, and optionally disintegrants and binders.
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