A pharmaceutical composition and its application, and a composition for treating prostate cancer and its application

Abstract

The present invention belongs to the technical field of tumor treatment, and particularly relates to a pharmaceutical composition and its application, as well as a composition for treating prostate cancer and its application. The pharmaceutical composition comprises a soluble copper salt and a copper ion carrier, wherein the copper ion carrier is at least one of ilisimol and disulfiram. The pharmaceutical composition can inhibit cancer cell proliferation and reduce the size of solid tumors. When used in combination with chemotherapy drugs at specific concentrations, it has a synergistic effect in inhibiting cancer cell proliferation and reducing cancer cell activity. The pharmaceutical composition can also inhibit chemotherapy-induced autophagy and improve chemotherapy drug resistance. Therefore, the pharmaceutical composition can enable chemotherapy drugs to produce ideal therapeutic effects even at a reduced dosage, thereby reducing the dosage of chemotherapy drugs and alleviating the side effects of chemotherapy without significantly increasing adverse reactions, providing a new combination drug regimen for tumor treatment.

Description

Technical Field

[0001] The present invention belongs to the technical field of tumor treatment, and in particular relates to a pharmaceutical composition and application thereof, and a composition for treating prostate cancer and application thereof. Background Art

[0002] Prostate cancer (PC) is the second leading cause of cancer-related death among men in the United States. By 2021, it is estimated that more than 240,000 new cases of PC will be diagnosed, and more than 34,000 men will die from the disease. Although the development of new treatments (including chemotherapy (docetaxel and cabazitaxel), immunotherapy (sipuleucel-T), bone-targeted therapy (radium-233 dichloride), and novel hormonal therapies (abiraterone acetate, enzalutamide, apalutamide, and darulutamide)) has improved the overall survival (OS) of patients with metastatic castration-resistant prostate cancer (mCRPC), this does not necessarily mean an improvement in the quality of life of these patients. The majority of mCRPC patients will eventually die from prostate cancer progression, so the search for better drugs to treat mCRPC is still in progress.

[0003] Since 2004, docetaxel has become the standard first-line chemotherapy for mCRPC. Docetaxel's cytotoxic effects primarily inhibit cancer cell mitosis and proliferation by arresting cells in the G2 and M phases. However, many tumors do not respond to it. Furthermore, due to the characteristics of malignant tumors, such as unlimited proliferation, infiltration and metastasis, metabolic abnormalities, and immune evasion, tumor cells exposed to the stressful environment of chemotherapeutic drugs gradually induce autophagy and develop drug resistance. This leads to drug resistance in most patients, resulting in suboptimal treatment efficacy. Even among patients who respond, drug-related adverse reactions are common, ultimately limiting the dosing and duration of docetaxel therapy. Insufficient intratumoral docetaxel concentrations may contribute to the reduced treatment outcomes. Other studies have shown that extended docetaxel treatment cycles (eight or more cycles versus six cycles) are associated with superior survival in prostate cancer patients. Summary of the Invention

[0004] To address the above technical issues, the present invention provides a pharmaceutical composition and its use, as well as a composition for treating prostate cancer and its use. The pharmaceutical composition can inhibit cancer cell proliferation and chemotherapy-induced autophagy, reducing chemotherapy drug resistance, thereby improving efficacy, reducing dosage, and alleviating chemotherapy side effects.

[0005] To achieve the above-mentioned purpose, the embodiment of the present invention adopts the following technical solutions:

[0006] In one aspect, the present invention provides a pharmaceutical composition comprising a soluble copper salt and a copper ion carrier, wherein the copper ion carrier is at least one of ilisimol and disulfiram.

[0007] Copper is a cofactor for essential enzymes, but homeostatic mechanisms within cells keep copper concentrations at very low levels to prevent the accumulation of free copper from damaging cells. Elesclomol (ES) and disulfiram are highly lipophilic small molecules that bind to copper ions. In mammalian cell lines, copper transport is defective and can be transferred across the membrane to key enzymes, thereby restoring mitochondrial function. Direct binding of copper to fatty acylated components of the tricarboxylic acid (TCA) cycle leads to the aggregation of fatty acylated proteins and the subsequent loss of iron-sulfur cluster proteins, resulting in proteotoxic stress and ultimately cell death. Experimental studies have confirmed that the pharmaceutical composition of the present invention can inhibit cancer cell proliferation and reduce the size of solid tumors.

[0008] The present invention also unexpectedly discovered during research that a pharmaceutical composition consisting of a soluble copper salt and a copper ion carrier, ilisimol, combined with the chemotherapy drug docetaxel, can significantly inhibit the proliferation and colony formation of prostate cancer cells and enhance their respective anti-prostate tumor activities. The two exhibit a significant synergistic effect at specific drug concentrations, which greatly enhances the efficacy of docetaxel. Furthermore, experimental studies have found that the pharmaceutical composition can inhibit autophagy induced by the chemotherapy drug docetaxel, thereby improving docetaxel resistance and enabling docetaxel to still produce the desired therapeutic effect at a reduced dosage, thereby reducing the dosage of docetaxel and alleviating the side effects of chemotherapy. Furthermore, the pharmaceutical composition does not significantly increase adverse reactions. When used in combination with docetaxel, it improves the efficacy of docetaxel while also not significantly increasing adverse reactions. Currently, there are no reports on the combined use of a soluble copper salt and a copper ion carrier composition with a chemotherapy drug such as docetaxel for therapeutic purposes, nor have there been reports on the use of a soluble copper salt and a copper ion carrier composition to inhibit chemotherapy-induced autophagy to improve chemotherapy drug intolerance.

[0009] The present invention provides a new combined drug regimen for the treatment of prostate cancer, which can improve the efficacy, greatly reduce the dosage of chemotherapy drugs, and alleviate the adverse reactions of patients.

[0010] In combination with the first aspect, the soluble copper salt is at least one of copper chloride and copper sulfate.

[0011] Optionally, the soluble copper salt is copper chloride, and the ion carrier is ilisimol.

[0012] Preferably, the molar ratio of copper chloride to ilisimol is 0.8 to 1.2:1.

[0013] Preferably, the molar ratio of copper chloride to ilisimol is 1:1.

[0014] In a second aspect, the present invention also provides use of the above-mentioned pharmaceutical composition in the preparation of a drug for treating prostate cancer.

[0015] In a third aspect, the present invention also provides the use of the above-mentioned pharmaceutical composition in the preparation of a drug for inhibiting chemotherapy-induced autophagy. This pharmaceutical composition can inhibit chemotherapy-induced autophagy, thereby improving chemotherapy drug resistance, allowing the chemotherapy drug to produce the desired therapeutic effect even at a reduced dosage, thereby reducing the dosage of the chemotherapy drug and alleviating the side effects of chemotherapy.

[0016] In combination with the third aspect, the chemotherapy drug is docetaxel.

[0017] In a fourth aspect, the present invention further provides a composition for treating prostate cancer, comprising docetaxel and the above-mentioned pharmaceutical composition.

[0018] The present invention has discovered that the combined use of the above-mentioned pharmaceutical composition and docetaxel can significantly enhance the original anti-prostate tumor activity of each, and the combined use of the two at specific drug concentrations exhibits a significant synergistic effect. Furthermore, the above-mentioned pharmaceutical composition can inhibit docetaxel-induced autophagy, thereby improving docetaxel resistance and enabling docetaxel to produce the desired therapeutic effect at a reduced dosage. This composition provides a novel and highly effective combined treatment option for cancer patients, alleviating the side effects of docetaxel while enhancing anti-tumor efficacy, thereby improving the prognosis and quality of life of cancer patients.

[0019] Preferably, the molar ratio of docetaxel to the pharmaceutical composition is 1 to 8:40.

[0020] Preferably, the molar ratio of docetaxel to the pharmaceutical composition is 1 to 2:40, at which the synergistic effect of the two is more significant.

[0021] More preferably, the molar ratio of docetaxel to the pharmaceutical composition is 1:40. At this molar ratio, the synergistic effect of the two is most significant.

[0022] In a fifth aspect, embodiments of the present invention further provide use of the above-mentioned composition in the preparation of a drug for treating prostate cancer.

[0023] Preferably, the drug is an injection, including injection solution, freeze-dried powder for injection, etc.

[0024] After the composition provided by the present invention is prepared into an injection, it can be administered by injection to treat prostate cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The cell viability after adding different concentrations of the copper chloride-elisimol composition in Example 19;

[0026] Figure 2 The cell viability after adding different concentrations of docetaxel and the combination of docetaxel and copper chloride-elisemol in Example 19;

[0027] Figure 3 The cell viability after adding 1 nM docetaxel and / or 20 nM copper chloride-elismol composition in Example 19;

[0028] Figure 4 The effect of the copper chloride-elisimol composition and its combination with docetaxel on the growth of PC3 cell clones in Example 20;

[0029] Figure 5 The effect of the copper chloride-elismol composition and its combination with docetaxel in Example 21 on the tumor volume in mice;

[0030] Figure 6 The effect of the copper chloride-elismol composition and its combination with docetaxel in Example 21 on tumor weight in mice;

[0031] Figure 7 The effect of the copper chloride-elismol composition and its combination with docetaxel in Example 21 on the body weight of mice;

[0032] Figure 8 The effect of the copper chloride-elismol composition and its combination with docetaxel in Example 21 on alanine aminotransferase (ALT) in mice;

[0033] Figure 9 The effect of the copper chloride-elismol composition and its combination with docetaxel in Example 21 on aspartate aminotransferase (AST) in mice;

[0034] Figure 10 The effect of the copper chloride-elismol composition and its combination with docetaxel in Example 21 on creatinine in mice;

[0035] Figure 11 The effect of the copper chloride-elismol composition and its combination with docetaxel in Example 21 on urea nitrogen (BUN) in mice;

[0036] Figure 12The effect of the copper chloride-elismol composition and its combination with docetaxel in Example 21 on mouse white blood cells (WBC);

[0037] Figure 13 This is the Western blot verification of the changes in the levels of the autophagy marker protein LC3 after the copper chloride-ilisimol composition and its combination with docetaxel in Example 22. DETAILED DESCRIPTION

[0038] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further illustrated below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0039] Unless otherwise specified, the raw materials and reagents used in the following examples were obtained from commercial sources.

[0040] Example 1

[0041] An embodiment of the present invention provides a pharmaceutical composition consisting of copper chloride and elisimol, wherein the molar ratio of copper chloride to elisimol is 1:1.

[0042] Example 2

[0043] An embodiment of the present invention provides a pharmaceutical composition consisting of copper chloride and elisimol, wherein the molar ratio of copper chloride to elisimol is 0.8:1.

[0044] Example 3

[0045] An embodiment of the present invention provides a pharmaceutical composition consisting of copper chloride and elisimol, wherein the molar ratio of copper chloride to elisimol is 1.2:1.

[0046] Example 4

[0047] An embodiment of the present invention provides a pharmaceutical composition consisting of copper sulfate and ilisimol, wherein the molar ratio of copper sulfate to ilisimol is 1:1.

[0048] Example 5

[0049] An embodiment of the present invention provides a pharmaceutical composition consisting of copper sulfate and disulfiram, wherein the molar ratio of copper sulfate to disulfiram is 1:1.

[0050] Example 6

[0051] An embodiment of the present invention provides a pharmaceutical composition consisting of copper chloride and disulfiram, wherein the molar ratio of copper chloride to disulfiram is 1:1.

[0052] Example 7

[0053] An embodiment of the present invention provides a pharmaceutical composition consisting of copper chloride, copper sulfate and ilisimol, wherein the molar ratio of copper chloride, copper sulfate and ilisimol is 1:1:2.

[0054] Example 8

[0055] An embodiment of the present invention provides a pharmaceutical composition consisting of copper chloride, copper sulfate, elisimol and disulfiram, wherein the molar ratio of copper chloride, copper sulfate, elisimol and disulfiram is 1:1:1:1.

[0056] Example 9

[0057] An embodiment of the present invention provides a composition for treating prostate cancer, which consists of docetaxel, copper chloride and elisimol, wherein the molar ratio of docetaxel, copper chloride and elisimol is 1:20:20.

[0058] Example 10

[0059] An embodiment of the present invention provides a composition for treating prostate cancer, which consists of docetaxel, copper chloride and elisimol, wherein the molar ratio of docetaxel, copper chloride and elisimol is 1:10:10.

[0060] Example 11

[0061] An embodiment of the present invention provides a composition for treating prostate cancer, which consists of docetaxel, copper chloride and elisimol, wherein the molar ratio of docetaxel, copper chloride and elisimol is 1:5:5.

[0062] Example 12

[0063] An embodiment of the present invention provides a composition for treating prostate cancer, which consists of docetaxel, copper chloride and elisimol, wherein the molar ratio of docetaxel, copper chloride and elisimol is 2:5:5.

[0064] Example 13

[0065] The present invention provides an embodiment of the pharmaceutical composition of Example 1 for use in preparing an injection for treating prostate cancer. The specific method is as follows: the pharmaceutical composition of Example 1 is prepared into an injection solution or a lyophilized powder for injection according to a conventional method according to the desired dosage form and pharmaceutically acceptable injection excipients.

[0066] This method can also be used for the pharmaceutical composition of any one of Examples 2 to 8.

[0067] Example 14

[0068] An embodiment of the present invention provides an injection for treating prostate cancer, comprising the pharmaceutical composition of any one of Examples 1 to 8 and a pharmaceutically acceptable injection excipient.

[0069] Example 15

[0070] The embodiments of the present invention provide a lyophilized powder for injection for treating prostate cancer, comprising the pharmaceutical composition of any one of Embodiments 1 to 8 and a pharmaceutically acceptable lyophilized powder excipient for injection.

[0071] Example 16

[0072] The present invention provides an embodiment of the use of the composition of Example 9 in preparing an injection for treating prostate cancer. The specific method is: the composition of Example 9 is prepared into an injection solution or a lyophilized powder for injection according to a conventional method according to the desired dosage form and pharmaceutically acceptable injection excipients.

[0073] This method can also be used for the pharmaceutical compositions obtained in Examples 10 to 12.

[0074] Example 17

[0075] An embodiment of the present invention provides an injection for treating prostate cancer, comprising the composition of any one of Examples 9 to 12 and a pharmaceutically acceptable injection excipient.

[0076] Example 18

[0077] The embodiments of the present invention provide a lyophilized powder for injection for treating prostate cancer, comprising the composition of any one of embodiments 9 to 12 and a pharmaceutically acceptable lyophilized powder excipient for injection.

[0078] Example 19

[0079] The present invention provides the pharmaceutical composition of Example 1 (hereinafter referred to as the copper chloride-elisimol composition) and its inhibitory effect on the proliferation of prostate cancer PC3 cells when used in combination with docetaxel.

[0080] Prostate cancer PC3 cell suspension was prepared using conventional 1640 culture medium containing 10% fetal bovine serum + 1% penicillin / streptomycin. After counting, cells were inoculated into 96-well plates, with approximately 100 μl (4,000 cells) per well. Four replicates were performed on the same sample.

[0081] A 96-well plate inoculated with a suspension of prostate cancer PC3 cells was pre-cultured in an incubator (37°C, 5% CO2). After the cells adhered and grew, they were incubated. When the cell number showed exponential growth, they were randomly divided into a control group and multiple drug-treated groups. The control group was treated with 1640 culture medium containing 10% fetal bovine serum and 1% penicillin / streptomycin, while each drug-treated group was treated with an equal volume of docetaxel and / or the copper chloride-elisemol combination (ES-Cu) solution of Example 1 at a predetermined concentration. After incubation for 48 hours, 100 μl of culture medium containing 10% CCK-8 was added to each well, and the cells were incubated in an incubator for 4 hours. The absorbance (OD) at 450 nm was then measured using a microplate reader.

[0082] The cell viability after adding different concentrations of the copper chloride-elisimol composition of Example 1 was calculated by absorbance, and the results were as follows: Figure 1 As shown in Table 1, the cell viability after adding different concentrations of docetaxel and adding different concentrations of docetaxel and 20mM copper chloride-ilisimol combination is shown in Table 2, Table 3 and Figure 2 shown.

[0083] Table 1 Cell viability after adding different concentrations of copper chloride-elisimol combination

[0084]

[0085] Table 2 Cell viability after adding different concentrations of docetaxel

[0086] Final docetaxel concentration (nM) 0 0.5 1 2 4 8 16 Cell viability (%) 99.5 96.0 56.0 32.7 16.1 8.1 7.1

[0087] Table 3 Cell viability after adding different concentrations of docetaxel and copper chloride-ilisimol combination

[0088]

[0089] From Tables 1 to 3 and Figure 1 、 Figure 2 It can be seen that when the final concentration of docetaxel was 0.5nM, the cell viability was 96.0%, which showed no significant decrease compared with the control group. When the concentration was increased, the cell viability decreased accordingly. This result is basically consistent with the recognized correlation between docetaxel concentration and its therapeutic effect. When the copper chloride-elismol composition with a final concentration of 20nM was added at the same concentration, the cell viability dropped significantly to 41.2%, and the cell viability at this time was much lower than the cell viability (about 65%) when only the 20μM / L copper chloride-elismol composition was added, indicating that the copper chloride-elismol composition and docetaxel have a synergistic effect in inhibiting the proliferation of prostate cancer cells at this concentration.

[0090] Similarly, when the final concentration of docetaxel was 1 nM to 4 nM, docetaxel alone reduced cell viability to 56.0%, 32.7%, and 16.1%, respectively. When a copper chloride-elisimol combination with a final concentration of 20 nM was added simultaneously, the cell viability should theoretically drop to 65% of the original value, that is, to 36.4%, 21.3%, and 10.5%, respectively. However, in reality, the cell viability dropped to 29.7%, 21.1%, and 9.2%, indicating that the copper chloride-elisimol combination and docetaxel also had a synergistic effect in inhibiting the proliferation of prostate cancer cells at the above concentrations. When the final concentration of docetaxel was 8 to 16 nM, no significant synergistic effect was shown in inhibiting the proliferation of prostate cancer cells.

[0091] A 1 nM docetaxel concentration was selected to validate the above results: 96-well plates seeded with a suspension of prostate cancer PC3 cells were pre-cultured in an incubator (37°C, 5% CO2). After the cells adhered and grew, the cells were incubated. When the cell number showed exponential growth, they were randomly divided into a control group and multiple drug-treated groups. The control group was treated with 1640 culture medium containing 10% fetal bovine serum and 1% penicillin / streptomycin, while each drug-treated group was treated with an equal volume of docetaxel and / or the copper chloride-ilisimol combination solution of Example 1 at a predetermined concentration. After incubation for 48 hours, 100 μl of culture medium containing 10% CCK-8 was added to each well, and the cells were incubated in an incubator for 1 hour. The absorbance (OD) at 450 nm was then measured using a microplate reader.

[0092] The cell viability after adding different concentrations of the copper chloride-elisimol composition of Example 1 was calculated by absorbance, and the results were as follows: Figure 3 As shown in Table 4, the cell viability after adding different concentrations of docetaxel and adding different concentrations of docetaxel and 20mM copper chloride-ilisimol combination is shown in Table 2, Table 3 and Figure 2 shown.

[0093] Table 4 Cell viability after addition of docetaxel and / or copper chloride-ilisimol combination

[0094]

[0095] From Table 4 and Figure 3 It can be seen that when the final concentration of docetaxel is 1 nM, docetaxel alone can reduce cell viability to 59.7%. When the copper chloride-elisimol composition with a final concentration of 20 nM is added at the same time, the cell viability should theoretically drop to 62.3% of the original value, that is, to 37.2%. In fact, the cell viability dropped to 34.3%. It can be seen that the copper chloride-elisimol composition and docetaxel do have a synergistic effect in inhibiting the proliferation of prostate cancer cells at the above concentrations.

[0096] Example 20

[0097] The present invention provides the pharmaceutical composition of Example 1 (hereinafter referred to as the copper chloride-elisimol composition) and its inhibitory effect on the colony formation of prostate cancer PC3 cells when used in combination with docetaxel.

[0098] A prostate cancer PC3 cell suspension was prepared in a conventional 1640 medium containing 10% fetal bovine serum + 1% penicillin / streptomycin, and plated in a 6-well plate at a density of 500 cells / well. After the cells adhered to the wall, they were cultured for 48 hours (cell number increased exponentially), and then replaced with fresh 1640 medium containing 10% fetal bovine serum + 1% penicillin / streptomycin. Culture medium or a preset concentration of docetaxel and / or the copper chloride-ilisimol composition of Example 1 were added and treated for 10 days. After aspirating the culture medium, 1 ml of 4% paraformaldehyde was added for 10 minutes, the waste liquid was removed, and the cells were washed twice with PBS. Then, 1 ml of crystal violet was added to each well for 10 minutes, and then washed with tap water until there was no staining. After drying in a 65°C oven, the colonies were counted. The drug concentrations of each sample are shown in Table 5.

[0099] Table 5 Drugs and their concentrations in each sample

[0100]

[0101] The results are as follows Figure 4 As shown, the combined use of the copper chloride-elisimol combination and docetaxel for 10 days significantly reduced the number of colonies, indicating that the combined use of the copper chloride-elisimol combination and docetaxel significantly inhibited the clone formation of prostate cancer PC3 cells.

[0102] Example 21

[0103] The present invention provides the pharmaceutical composition of Example 1 (hereinafter referred to as the copper chloride-elisimol composition) and its inhibitory effect on tumor growth in vivo when used in combination with docetaxel.

[0104] First, 6-8 week-old male C57BL / 6 mice were subcutaneously injected with 2×106 RM-1 mouse prostate cancer cells into the flank of the forelimb to establish a mouse tumor-bearing model. The mice were then randomly divided into a control group, a copper chloride-elismomol combination treatment group, a docetaxel treatment group, and a combination treatment group. The copper chloride-elismomol combination treatment group received an intraperitoneal injection of 0.5 mg / kg copper chloride and 1.5 mg / kg elismomol; the docetaxel treatment group received an intraperitoneal injection of 2 mg / kg docetaxel; the combination treatment group received an intraperitoneal injection of 0.5 mg / kg copper chloride, 1.5 mg / kg elismomol, and 2 mg / kg docetaxel; and the control group received normal saline. The dosing volume for each group was 100 μl. All groups received intraperitoneal injections starting on day 4 after tumor implantation and every three days. Body weight and tumor volume were measured every three days. The drug was injected 6 times in total. The liver and kidney function and blood routine of the mice were measured 24 hours after the last administration. The mice were killed and the tumors were removed and weighed. The changes in tumor volume of mice in each group are shown in Table 6 and Figure 5 The tumor weights are shown in Tables 7 and Figure 6 shown.

[0105] Table 6 Tumor volume of mice in each group (mm 3 )

[0106]

[0107] Table 7 Tumor weight of mice in each group (g)

[0108]

[0109] The results showed that the tumor volume of the groups treated with either the copper chloride-elisemol combination alone or docetaxel was smaller than that of the control group, while the tumor volume of the combined treatment group was significantly reduced, indicating that the combined treatment of the copper chloride-elisemol combination and docetaxel significantly enhanced the tumor inhibitory effects of both.

[0110] The body weights of mice in each group are shown in Table 8 and Figure 7 As shown, the results of liver and kidney function and blood routine tests are as follows Figures 8-12 shown.

[0111] Table 8 Body weight of mice in each group (g)

[0112]

[0113] From Table 8 and Figures 7-12 The results show that the copper chloride-ilisimol combination does not significantly increase the body's adverse reactions, and does not significantly increase the body's adverse reactions when used in combination with docetaxel.

[0114] Example 22

[0115] The present invention provides the pharmaceutical composition of Example 1 (hereinafter referred to as the copper chloride-ilisimol composition) and its combined use with docetaxel in improving docetaxel resistance by inhibiting docetaxel-induced autophagy.

[0116] This experiment verified changes in the levels of the autophagy marker protein LC3 by Western blot. The experimental culture medium used was 1640 medium containing 10% fetal bovine serum and 1% penicillin / streptomycin. The specific experimental steps were as follows:

[0117] PC-3 cells were seeded into four 25 cm² cell culture flasks and cultured in an incubator maintained at 37°C and 5% CO². After cells adhered and grew to a density of 60%-70%, they were randomly divided into a control group, a copper chloride-elisemol combination group, a docetaxel group, and a combination treatment group. The culture medium was changed routinely. The copper chloride-elisemol combination group received the copper chloride-elisemol combination at a final concentration of 20 nM, the docetaxel group received docetaxel at a final concentration of 1 nM, and the combination treatment group received both the copper chloride-elisemol combination and docetaxel at a final concentration of 20 nM copper chloride-elisemol and 1 nM docetaxel. After incubation for 48 hours, total cell protein was extracted. Equal amounts of protein extract were taken from each group, made up to volume with RIPA lysis buffer, mixed thoroughly with 5X loading buffer, and denatured in a boiling water bath for 5 minutes. A 5% stacking gel and resolving gels of varying concentrations were prepared. Equal amounts of samples and ladder markers were added to the sample wells of the gel, and electrophoresis and semi-dry transfer were started. The membrane was blocked with 5% skim milk powder for 2 hours. After the primary antibody was bound overnight, the secondary antibody was blocked for 2 hours, and the antibody-specific binding bands were detected using a chemiluminescence instrument with automatic exposure. Figure 13 As shown in the results, the copper chloride-ilisimol combination can inhibit docetaxel-induced autophagy, thereby improving docetaxel resistance.

[0118] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A composition for treating prostate cancer, characterized in that: The invention comprises docetaxel and a pharmaceutical composition; wherein the pharmaceutical composition is composed of a soluble copper salt and a copper ion carrier, the soluble copper salt is copper chloride, and the ion carrier is ilisimol; The molar ratio of the copper chloride to ilisimol is 0.8-1.2:

1.

2. The composition for treating prostate cancer according to claim 1, characterized in that The molar ratio of the copper chloride to ilisimol is 1:

1.

3. The composition for treating prostate cancer according to claim 1, characterized in that The molar ratio of the docetaxel to the pharmaceutical composition is 1-8:

40.

4. Use of the composition according to any one of claims 1 to 3 in the preparation of a medicament for treating prostate cancer.

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