CN111603459A

CN111603459A - Application of suramin compound in preparation of phosphoinositide kinase inhibitor

Info

Publication number: CN111603459A
Application number: CN202010618298.8A
Authority: CN
Inventors: 饶枫; 张晓哲; 石少东
Original assignee: Southwest University of Science and Technology
Current assignee: Southwest University of Science and Technology; Southern University of Science and Technology
Priority date: 2020-06-30
Filing date: 2020-06-30
Publication date: 2020-09-01

Abstract

The invention provides an application of suramin compounds in preparation of a pentaphosphoinositide kinase inhibitor. The invention provides a new application of suramin compounds, which can be used as a pentaphosphoinositide kinase inhibitor and can inhibit the activity of pentaphosphoinositide kinase, and the binding constant kd of IP5K and suramin is 1.1 mu M, IC₅₀It was 2.98. mu.M. Meanwhile, suramin compounds are used as a pentaphosphoinositide kinase inhibitor and are combined with MLN4924, so that the anti-tumor effect of the MLN4924 can be enhanced, the using amount of the MLN4924 is reduced, the non-specific toxic and side effects are relieved, and the biological safety is improved.

Description

Application of suramin compound in preparation of phosphoinositide kinase inhibitor

Technical Field

The invention belongs to the field of biological medicines, and particularly relates to an application of suramin compounds in preparation of a pentaphosphoinositide kinase inhibitor.

Background

In human diseases, tumors are a big problem to be overcome urgently, and at present, people are always searching and exploring medicaments for treating tumors. Inositol hexaphosphate (IP6) is a natural signal small molecule in vivo that acts as a "molecular glue" between the ubiquitin ligase CullinRING E3Ligases (CRL) and its inhibitory complex COP9 signalosome (csn), where CRL is dynamically regulated by pseudomorphic modification (Neddylation). IP6 binds to CSN and promotes depsipylation (denedylation) of CRL ubiquitin ligase, participates in signal transduction of cells, and enables tumor cells to survive.

The synthetase of IP6 is a pentaphosphoinositide kinase (IP 5K). As a small molecule kinase, the phosphoinositide kinase has the approximate structural domain of the traditional kinase, but the binding pocket of the substrate is greatly different from the traditional kinase, and an inhibitor with good specificity and small side effect is expected to be designed. At present, no inhibitor specifically targeting phosphoinositide kinase has been reported.

Therefore, providing an inhibitor of phosphoinositide kinase to reduce the synthesis of IP6 is of great significance in inhibiting signal transduction between cells, promoting apoptosis of tumor cells, and retarding cell cycle progression.

MLN4924 as an inhibitor for inhibiting E3 ubiquitin enzyme paranoid, is a potent and selective NEDD8 activating enzyme (NAE) inhibitor, and is the only clinically used paranoid enzyme inhibitor at present. In HCT-116 cells, MLN4924 resulted in a decrease in Ubc12-NEDD8 thioester and NEDD 8-arrestin conjugates, thereby inhibiting CRL-ubiquitination-mediated protein degradation (such as CDT1 protein), while accumulation of CDT1 protein caused cell cycle defects. Currently, MLN4924 has been used in phase III clinical trials for clinical tumor therapy, and has some role in cancer therapy, but has greater toxicity.

Suramin is a symmetrical polysulfonate naphthylurea drug, and the structure of suramin is shown as a formula I.

Suramin is known to be an anti-trypanosome and antiviral drug at present, and no report that suramin can be used as an inhibitor of phosphoinositide kinase exists, and no clear result shows that suramin has an anti-tumor effect.

Disclosure of Invention

In view of the problems in the prior art, the invention aims to provide the application of suramin compounds in preparing phosphoinositide kinase inhibitors.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the invention provides the use of a Suramin (Suramin) compound in the preparation of a pentaphosphoinositide kinase inhibitor.

The invention provides a new application of suramin compounds as phosphoinositide kinase inhibitors.

Preferably, the suramin compound comprises any one of suramin, suramin derivatives or suramin pharmaceutically acceptable salts. The suramin derivatives include NF449 and NF 110.

Wherein, the structure of NF449 is shown as formula II:

the structure of NF110 is shown in formula III:

the suramin compounds can strongly inhibit the activity of pentaphosphoinositide kinase (IP 5K). Wherein suramin inhibits IC50 of IP5K at 2.8 μ M, NF449 inhibits IC50 of IP5K at 1.1 μ M, and NF110 inhibits IC50 of IP5K at 3.1 μ M.

The IP5K is a synthetase of IP6, and the strong inhibition of IP5K can reduce the synthesis of IP6 and further regulate and control a downstream target of IP 6. The reduction of IP6 synthesis is beneficial to blocking the combination of ubiquitin ligase and its inhibitory complex, retarding cell cycle progression and promoting apoptosis of tumor cells.

In a second aspect, the present invention provides an anti-tumor pharmaceutical composition comprising a suramin compound and a peptidomimetic enzyme inhibitor.

As a preferred embodiment of the present invention, the paranoid enzyme inhibitor comprises MLN 4924.

Preferably, the suramin compound comprises any one of suramin, suramin derivatives or suramin pharmaceutically acceptable salts.

Preferably, the suramin derivative comprises NF449 and NF 110.

Suramin (Suramin) and Suramin derivatives NF449 and NF110 are effective in inhibiting pentaphosphoinositide kinase in vitro and in vivo and in turn modulate downstream targets of IP 6. Suramin was found to promote sumiins were treated with suramin, and to reduce the binding of CSN2 to Cullin4A, while detecting substrate receptors and substrates downstream of CRL ubiquitin ligase, suramin was found to promote the degradation of substrate receptors and substrates. MLN4924 as an inhibitor for inhibiting E3 ubiquitinase enzymatic sumoylation has a certain effect in cancer treatment, and has entered phase III clinical trials, but has great toxicity.

In the invention, the combined application of suramin and MLN4924 is found to promote the apoptosis of HCT116, Hela and other tumor cells, retard the cell cycle progress and achieve the purposes of inhibiting the growth of the tumor cells and treating tumors. And the combined medicament has better biological safety, reduces the cytotoxicity of a single medicament, and provides a new strategy and thought for treating tumors.

As a preferable technical scheme, the pharmaceutical composition is a single compound preparation.

As a preferred technical scheme, the pharmaceutical composition is a combination of two separate preparations, namely a suramin compound preparation and a bacteroid enzyme inhibitor.

Preferably, the two separate formulations are administered simultaneously.

Preferably, the two separate formulations are administered sequentially.

The pharmaceutical composition can be in the form of a single compound preparation, and can also be a combination of two separate preparations; when the two separate preparations are combined, the administration mode can be simultaneous administration or sequential administration, for example, the suramin compound can be administered first, the MLN4924 can be administered after a certain time interval, the MLN4924 can be administered first, the suramin compound can be administered after a certain time interval, or the suramin compound and the MLN4924 can be administered alternately.

In the invention, the using concentration of the suramin compound and the MLN4924 can be adjusted according to the specific disease condition of a patient, and the optimal treatment concentration and using ratio are selected.

As a preferable technical scheme, the pharmaceutical composition further comprises pharmaceutically acceptable auxiliary materials.

Preferably, the auxiliary materials comprise any one or a combination of at least two of a carrier, a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrating agent, an emulsifier, a cosolvent, a solubilizer, an osmotic pressure regulator, a surfactant, a coating material, a coloring agent, a pH regulator, an antioxidant, a bacteriostatic agent or a buffering agent.

Preferably, the dosage form of the pharmaceutical composition is any one of common compressed tablets, dispersible tablets, enteric-coated tablets, capsules, granules, dripping pills, emulsions, powders, oral liquids or injections.

Preferably, the administration route of the pharmaceutical composition comprises any one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or transdermal administration or a combination of at least two of the same.

In a third aspect, the use of the pharmaceutical composition according to the second aspect in the preparation of an anti-tumor medicament.

In a fourth aspect, the present invention also provides a novel anti-tumor combination therapy which is a combination therapy of suramin-like compounds and MLN 4924.

The suramin compound and the MLN4924 are combined, so that the composition has stronger anti-tumor activity and higher biological safety compared with a single conventional medicament MLN4924, can kill tumor cells more effectively, and provides a new strategy and thought for tumor treatment; in addition, the combination therapy can obviously inhibit the activity of the pentaphosphoinositide kinase, inhibit the synthesis of IP6 and exert more effective anticancer activity.

The recitation of numerical ranges herein includes not only the above-recited values, but also any values between any of the above-recited numerical ranges not recited, and for brevity and clarity, is not intended to be exhaustive of the specific values encompassed within the range.

Compared with the prior art, the invention has at least the following beneficial effects:

(1) the invention provides a new application of suramin compounds, wherein suramin can be used as a pentaphosphoinositide kinase inhibitor, the binding constant kd of IP5K and suramin is 1.1 mu M, and the IC50 is 2.8 mu M; meanwhile, suramin can inhibit the combination of IP5K and substrate inositol pentaphosphate, and the combination constant of the inositol pentaphosphate and IP5K is increased from 1.5 mu M to 9.5 mu M in the presence of suramin; so that the inositol hexaphosphate can not be normally synthesized and can not exert the adhesive action between the ubiquitin ligase and the inhibition complex;

(2) the anti-tumor pharmaceutical composition provided by the invention uses suramin compounds as a phosphoinositide kinase inhibitor and is combined with a paranoid enzyme inhibitor MLN4924, and the suramin compounds can promote tumor cell apoptosis caused by MLN4924 and further inhibit the growth and proliferation of tumor cells; therefore, the suramin compound can be used as a cooperative drug of MLN4924 to treat tumors, so that the dosage of MLN4924 is reduced, the non-specific toxic and side effects are relieved, and the biological safety is improved.

Drawings

FIG. 1 is a Native PAGE gel detection of IP5 and IP6 content at different suramin concentrations in example 1.

FIG. 2 is a graph of suramin concentration versus conversion of IP6/IP5 in example 1.

FIG. 3 is a graph of the original plot and the corresponding fitted curve obtained when isothermal titration calorimetry was used to detect the interaction of IP5K with suramin in example 2.

FIG. 4 is a graph of the original pattern and corresponding fit obtained in example 2 using isothermal titration calorimetry to detect interaction of IP5K with suramin in combination with IP 5.

FIG. 5 is a graph of the original plot and the corresponding fitted curve obtained when isothermal titration calorimetry was used to detect the interaction of IP5K with IP5 in example 2.

FIG. 6(a) is a Native PAGE detection gel of example 3 after suramin treatment of HCT116 cells.

FIG. 6(b) is a bar graph of the amount of intracellular IP6 in treated and untreated suramin according to the glue map analysis in example 3.

FIG. 7 is a bar graph of cell viability following treatment with different concentrations of MLN4924 and suramin in example 4.

FIG. 8(a) is a graph showing the results of cell cycle measurement of each experimental group after PI staining in example 5.

FIG. 8(b) is a graph showing the ratio of the cell phases in each experimental group in example 5.

FIG. 8(c) is a graph showing the results of detecting apoptosis using western blot in each experimental group in example 5.

FIG. 8(d) is a graph showing the results of the flow-through experiments in each experimental group after double staining with Annexin-V and PI in example 5.

FIG. 9 is a Native PAGE gel detection of IP5 and IP6 content at different NF449 concentrations in example 6.

FIG. 10 is a graph of NF449 concentration versus IP6/IP5 conversion in example 6.

FIG. 11 is a bar graph of cell viability after treatment with different concentrations of MLN4924 and NF449 in example 6.

FIG. 12 is a Native PAGE gel detection of IP5 and IP6 content at different NF110 concentrations in example 7.

FIG. 13 is a graph of NF110 concentration versus IP6/IP5 conversion in example 7.

FIG. 14 is a bar graph of cell viability following treatment with different concentrations of MLN4924 and NF110 in example 7.

Detailed Description

The technical solutions of the present invention are further described in the following embodiments with reference to the drawings, but the following examples are only simple examples of the present invention and do not represent or limit the scope of the present invention, which is defined by the claims.

In the following examples, reagents and instruments used are available from conventional sources unless otherwise specified, and the experimental procedures used are those known to those skilled in the art.

Example 1

This example is used to demonstrate that suramin has an inhibitory effect on phosphoinositide kinase.

An E.coli expression plasmid of human-derived pentaphosphoinositide kinase (IP5K) was first constructed by an experimental method commonly used in the art, and human-derived pentaphosphoinositide kinase was expressed in E.coli.

The phosphoinositide kinase is phosphoinositide kinase which can convert the phosphoinositide (IP5) into the inositol hexaphosphate (IP6), according to the characteristic, suramin solutions with the concentrations of 0, 0.5 mu M, 1 mu M, 2 mu M, 4 mu M, 8 mu M, 12.5 mu M and 25 mu M are respectively prepared and then mixed with IP5K and IP5, and the inhibition effect of suramin on the generation of IP6 in the enzyme activity reaction is detected by Native PAGE gel.

As shown in a figure 1 of Native PAGE gel detection results, with the increase of suramin concentration, the content of IP6 is gradually reduced, namely IP5K cannot continuously catalyze the conversion of IP5 to IP6, which indicates that suramin inhibits the effect of IP5K and is an effective pentaphosphoinositide kinase inhibitor.

FIG. 2 is a graph of suramin concentration versus conversion of IP6/IP5 plotted according to Native PAGE gel detection results, wherein the abscissa of the graph is suramin concentration (. mu.M) and the ordinate is conversion/% (turnover), thus obtaining suramin IC₅₀At 2.8. mu.M.

Example 2

The binding constant of suramin to IP5K was determined using an Isothermal Titration Calorimetry (ITC) experiment in this example.

ITC is a thermodynamic technique for monitoring any chemical reaction initiated by the addition of a binding component, and has been the method of choice for identifying biomolecular interactions. The calorimetric curve of a change process is continuously and accurately monitored and recorded by a high-sensitivity and high-automation micro calorimeter, and thermodynamic and kinetic information is provided in situ, on line and without damage.

The ITC monitoring results are shown in fig. 3 to 5. Among them, fig. 3 demonstrates that suramin can bind to IP5K with binding constants kd ═ 1.1 ± 0.2 μ M, and N ═ 1.4 ± 0.1. Suramin was also shown to inhibit IP5 from binding to IP 5K.

Meanwhile, as can be seen from fig. 4, when suramin is bound to IP5K, the binding constant kd between IP5 and IP5K is 9.5 μ M, while as can be seen from fig. 5, when suramin is not present, the binding constant kd between IP5 and IP5K is 1.5 μ M, which indicates that suramin significantly inhibits the binding between IP5 and IP5K, and thus cannot be converted into IP 6.

Example 3

This example demonstrates the inhibitory effect of suramin on IP5K in cells.

Treating colon cancer cells HCT116 with 20. mu.M suramin, collecting HCT116 cells after 6 hours to extract IP6, as shown in FIG. 6(a) and FIG. 6 (b);

the Native PAGE detection in FIG. 6(a) shows that the content of IP6 in HCT116 cells is significantly reduced compared to the control group treated with suramin at 0. mu.M;

in fig. 6(b), the content of the experimental IP6 is only 60% based on 100% of the content of the control IP6, which is significantly reduced, indicating that suramin can significantly inhibit the activity of the intracellular IP5K protein.

Example 4

This example demonstrates that suramin and MLN4924 in combination have significant inhibitory effects on HCT 116.

HCT 11624 hours was treated with combinations of suramin and MLN4924 at concentrations of 0. mu.M, 20. mu.M and 50. mu.M, respectively, and MLN4924 at concentrations of 0. mu.M, 0.2. mu.M and 0.5. mu.M, respectively, and the results are shown in FIG. 7;

as can be seen from FIG. 7, when the concentration of MLN4924 is 0 μ M, i.e., when treated with suramin alone, there is no significant effect on the viability of the cells, and when the concentration of suramin is 20 μ M or 50 μ M and the concentration of MLN4924 is 0.2 μ M or 0.5 μ M, the viability of the tumor cells is significantly different from that when suramin is not used, especially when the concentration of suramin is 50 μ M, the inhibition effect on the tumor cells is most significant after being matched with MLN 4924.

The above examples demonstrate that MLN4924 in combination with suramin can promote apoptosis of tumor cells, retard growth of tumor cells,

example 5

This example was used to study the mechanism by which suramin promotes the killing of tumor cells by MLN 4924.

In order to study the mechanism of Suramin in promoting the killing of tumor cells by MLN4924, in this example, MLN4924 alone (designated as MLN4924 group), Suramin alone (designated as Suramin group), and a combination of Suramin and MLN4924 (designated as MLN4924+ Suramin group) were used to treat HCT 11624 hours, collect cells, and detect the apoptotic pathway proteins of HCT116 cells after drug treatment, and simultaneously detect the cell cycle.

(1) Detecting the cell cycle by using a PI staining method;

PI (propidium iodide) is a DNA dye, which is impermeable to cell membranes, and is not able to stain normal and apoptotic cells, while staining necrotic cells.

The results are shown in FIG. 8(a), which shows the results of cell cycle measurements of the blank control group (untreated cells), the MLN4924 group, the Suramin group, and the MLN4924+ Suramin group, respectively, and it can be seen that Suramin aggravates the cell cycle arrest of HCT116 by MLN 4924;

FIG. 8(b) shows data obtained from flow cytometry analysis, wherein G1 shows the G1 phase of the cell cycle interphase, which is the pre-DNA synthesis phase, S shows the S phase of the cell cycle interphase, which is the DNA synthesis phase, and G2/M shows the G2/M phase of the cell cycle interphase, which is the post-DNA synthesis phase; in the MLN4924+ Suramin group, there were significantly more cells in the late stage of DNA synthesis and fewer cells in the G1 stage.

(2) western blot detects apoptosis-related proteins clear-caspase 3 and PARP;

the results are shown in FIG. 8(c), FIG. 8(c) shows the western blot detection results obtained after treating the cells of each group, the data on the left side of the graph shows the protein band size, and the bands from top to bottom (I-VI) in the graph respectively show RARP I.e., cleared-RARP, RARP.e., cleared-RARP, cleared-Caspase 3 and actin, and it can be seen that the treatment of the cells with suramin and MLN4924 results in the increase of cleaved bands of Caspase3 and cleaved bands of PARP, which indicates that the combined administration of two drugs promotes the apoptosis of HCT116 cells;

(3) detecting apoptosis by Annexin-V and PI double staining method;

the results are shown in fig. 8(d), and it can be seen from the flow test that the ratio of apoptotic cells increases when the cells are treated with the combination, and the increase is more significant than that when MLN4924 is used alone, and the test results fully indicate that suramin promotes the apoptosis caused by MLN 4924.

Example 6

This example is used to demonstrate that NF449 has an inhibitory effect on phosphoinositide kinase and that the combination of NF449 and MLN4924 inhibits the proliferation of HCT116 cells.

NF449 solutions with the concentrations of 0, 0.5 muM, 1 muM, 2 muM, 4 muM, 8 muM, 12.5 muM and 25 muM are respectively prepared and then mixed with IP5K and IP5, and the inhibition effect of NF449 on the generation of IP6 in the enzyme activity reaction is detected by Native PAGE gel.

The Native PAGE gel detection result is shown in FIG. 9, the generation amount of IP6 is gradually reduced along with the increase of the concentration of NF449, namely IP5K cannot continuously catalyze the conversion of IP5 to IP6, and the result shows that NF449 inhibits the effect of IP5K and is a potent pentaphosphoinositide kinase inhibitor.

FIG. 10 is a graph of NF449 concentration plotted against the conversion of IP6/IP5 plotted against Native PAGE gel, wherein the abscissa is suramin concentration (. mu.M) and the ordinate is conversion/% (turnover), giving an IC50 of suramin of 1.1. mu.M.

The HCT 11624 hours was treated with a combination of NF449 and MLN4924, where the concentration of NF449 was 10 μ M and the concentration of MLN4924 was 0 μ M, 0.2 μ M, 0.5 μ M and 1 μ M, respectively, and the results are shown in FIG. 11, in which the left side of each set of data is a data graph of a control group without added NF 449;

when the concentration of the MLN4924 is 0 μ M, namely the treatment with NF449 alone, the activity of the cells is not obviously influenced, and when the concentration of the MLN4924 is 0.2 μ M, 0.5 μ M or 1 μ M, the activity of the tumor cells is gradually reduced under the condition of the combination of NF449 and MLN4924, which shows that the inhibition effect of the NF449 and the MLN4924 on the tumor cells is most obvious.

Thus, this example demonstrates that MLN4924 in combination with NF449 can block tumor cell growth in the treatment of tumors.

Example 7

This example is used to demonstrate that NF110 has inhibitory effects on phosphoinositide kinase and that the combination of NF110 and MLN4924 inhibits proliferation of HCT116 cells.

NF110 solutions with the concentrations of 0, 0.5 mu M, 1 mu M, 2 mu M, 4 mu M, 8 mu M, 12.5 mu M and 25 mu M are respectively prepared and then mixed with IP5K and IP5, and the inhibition effect of NF4110 on the generation of IP6 in the enzyme activity reaction is detected by Native PAGE gel.

The Native PAGE gel detection result is shown in FIG. 12, the generation amount of IP6 is gradually reduced along with the increase of the concentration of NF110, namely IP5K cannot continuously catalyze the conversion of IP5 to IP6, and the result shows that the NF110 inhibits the action of IP5K, is an effective pentaphosphoinositide kinase inhibitor, but the inhibition effect is weaker than that of suramin.

FIG. 13 is a graph of NF110 concentration and conversion of IP6/IP5 plotted according to Native PAGE gel detection results, wherein the abscissa of the graph is NF110 concentration (. mu.M) and the ordinate is conversion/% (turnover), and the IC50 of suramin is 3.5. mu.M.

The HCT 11624 hours was treated with a combination of NF110 and MLN4924, in which the concentration of NF110 was 20. mu.M and the concentration of MLN4924 was 0. mu.M, 0.5. mu.M and 1. mu.M, respectively, and the results are shown in FIG. 14, in which the left side of each set of data is a data graph of a control group without NF110 added;

when the concentration of the MLN4924 is 0 mu M, namely the treatment with the NF110 alone has no obvious influence on the viability of the cells, and when the concentration of the MLN4924 is 0.5 mu M or 1 mu M, the viability of the tumor cells is gradually reduced under the condition of the combined use of the NF110 and the MLN4924, which shows that the inhibition effect of the NF110 and the MLN4924 on the tumor cells is obvious after the cooperation.

Thus, this example demonstrates that MLN4924 in combination with NF110 can block tumor cell growth when used to treat tumors.

In conclusion, the suramin compound is an IP5K inhibitor, can influence the generation of IP6 in cells by inhibiting the activity of IP5K, and can promote the apoptosis of tumor cells and inhibit the cell cycle by combining with MLN4924, thereby improving the killing effect of MLN4924 on the tumor cells.

The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be understood by those skilled in the art that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are within the scope and disclosure of the present invention.

Claims

1. An application of suramin compounds in preparing phosphoinositide kinase inhibitor is provided.

2. The use according to claim 1, wherein the suramin-based compound comprises any one of suramin, suramin derivatives or suramin pharmaceutically acceptable salts.

3. An anti-tumor pharmaceutical composition, which comprises suramin compounds and a paranoid enzyme inhibitor.

4. The anti-tumor pharmaceutical composition according to claim 3, wherein the paradoximes inhibitor comprises MLN 4924;

preferably, the suramin compound comprises any one of suramin, suramin derivatives or suramin pharmaceutically acceptable salts;

preferably, the suramin derivative comprises NF449 and/or NF 110.

5. The anti-tumor pharmaceutical composition according to claim 3 or 4, wherein the pharmaceutical composition is a single compound preparation.

6. The anti-tumor pharmaceutical composition according to claim 3 or 4, wherein the pharmaceutical composition is a combination of two separate preparations, namely a suramin compound preparation and a bacteroid kinase inhibitor;

preferably, the two separate formulations are administered simultaneously;

preferably, the two separate formulations are administered sequentially.

7. The anti-tumor pharmaceutical composition according to any one of claims 3 to 6, further comprising pharmaceutically acceptable excipients;

preferably, the auxiliary materials comprise any one or a combination of at least two of a carrier, a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrating agent, an emulsifier, a cosolvent, a solubilizer, an osmotic pressure regulator, a surfactant, a coating material, a coloring agent, a pH regulator, an antioxidant, a bacteriostatic agent or a buffering agent;

preferably, the dosage form of the pharmaceutical composition is any one of ordinary compressed tablets, dispersible tablets, enteric-coated tablets, capsules, granules, dripping pills, emulsions, powder, oral liquid or injections;

8. Use of a pharmaceutical composition according to any one of claims 3 to 7 in the preparation of an anti-tumor medicament.

9. Use of a pharmaceutical composition according to any one of claims 3 to 7 in the preparation of a cell cycle retardant.

10. Use of a pharmaceutical composition according to any one of claims 3 to 7 in the preparation of an apoptosis-promoting agent.