WO2024120523A1 - Pharmaceutical composition for treating prostate cancer and use thereof - Google Patents

Abstract

The present disclosure provides a pharmaceutical composition for treating prostate cancer and use thereof. The pharmaceutical composition comprises (a) compound X or a pharmaceutically acceptable salt thereof; and (b) an androgen receptor antagonist or a pharmaceutically acceptable salt thereof. The present disclosure further provides a drug for treating prostate cancer, the drug comprising the pharmaceutical composition. Research has found that compound X and the androgen receptor antagonist have synergistic effects when used in combination for treating prostate cancer, which can better inhibit the proliferation of prostate cancer cells and promote the apoptosis of prostate cancer cells. The present disclosure demonstrates, by means of experiments, that the potential value of compound X used in combination with the androgen receptor antagonist, proving that the efficacy when the two are used in combination is much greater than that when they are used alone, providing a superior therapeutic regimen for treating prostate cancer, and having broad application prospects.

Description

A drug combination for treating prostate cancer and its application

Citation of Related Applications

This disclosure claims the priority of the invention patent application filed with the Patent Office of China on December 9, 2022, with application number 202211585939.X and invention name “A drug combination for treating prostate cancer and its application”, and the invention patent application filed with the Patent Office of China on December 1, 2023, with application number 202311643355.8 and invention name “A drug combination for treating prostate cancer and its application”, the entire contents of which are incorporated into this disclosure by reference.

Technical Field

The present invention belongs to the field of biomedicine technology, and specifically relates to a drug combination for treating prostate cancer and application thereof.

Background technique

Prostate cancer refers to an epithelial malignant tumor that occurs in the prostate. In 2004, WHO's "Pathology and Genetics of Tumors of the Urinary System and Male Reproductive Organs" included adenocarcinoma (acinar adenocarcinoma), ductal adenocarcinoma, urothelial carcinoma, squamous cell carcinoma, and adenosquamous carcinoma in terms of pathological types of prostate cancer. Among them, prostate adenocarcinoma accounts for more than 95%, so what we usually call prostate cancer refers to prostate adenocarcinoma. At present, prostate cancer has become an important disease that threatens the health of elderly men. Its incidence rate is high in Western countries and is increasing year by year. In Asian countries, where the incidence rate was low in the past, the number of patients has also increased rapidly in recent years. Commonly used methods for clinical treatment of prostate cancer include surgical resection, radiotherapy, and endocrine therapy that blocks androgens. Androgens are closely related to the growth of the prostate and the occurrence of prostate cancer, so endocrine therapy has become an effective way to treat prostate cancer. This method includes orchiectomy, estrogen therapy, gonadotropin-releasing hormone analog therapy, gonadotropin-releasing hormone antagonist therapy, androgen antagonist therapy, etc. Among them, androgen antagonist therapy can be used alone to treat early prostate cancer or as an adjuvant therapy in conjunction with surgery. It is one of the main means of clinical treatment of prostate cancer. As the target of androgen's biological effects, androgen receptor has become a research hotspot in the biomedical field. important content.

The androgen receptor (AR) belongs to the nuclear receptor superfamily and contains 918 amino acid residues, which form three important domains, namely the DNA binding domain (DBD), the ligand binding domain (LBD) and the N-terminal domain (NTD). Clinical experiments have shown that the administration of exogenous androgens to patients with prostate cancer will cause the patient's condition to worsen; on the contrary, if the testicles are removed and the androgen level in the patient's body is reduced, the condition will be alleviated, which shows that androgens have an important influence on the development of prostate cancer. According to the receptor theory, androgens must bind to AR to cause subsequent physiological and pathological effects, which lays a theoretical foundation for the use of androgen receptor antagonists to treat prostate cancer. Androgen receptor inhibition is one of the treatment strategies provided for patients with prostate cancer.

Compound X is a dual-ligand small molecule drug conjugate. The two branches of the dual ligand specifically recognize the folate receptor α and the prostate specific membrane antigen (PSMA) receptor, respectively. The small molecule toxin is Monomethyl auristatin E (MMAE), and the two are coupled through a linker. The linker is relatively stable in the extracellular fluid. After compound X is internalized by tumor cells, the linker can be cleaved by tissue proteases and MMAE is released. MMAE can bind to microtubules, causing the cell differentiation cycle to stagnate in the G2/M cycle, thereby inducing cell apoptosis and exerting an anti-tumor effect.

PSMA is highly and specifically expressed on the surface of prostate cancer cells, and has extremely important research value in the field of molecular imaging and targeted therapy of prostate cancer. In particular, radionuclide-labeled PSMA small molecule compounds have shown better clinical application prospects than traditional methods. In addition, PSMA is also highly expressed in tumor tissues or tumor neovascularization of other solid tumors such as renal cell carcinoma and squamous cell carcinoma of the lung. Currently, there are many drugs targeting PSMA in the early clinical development stage. Folic acid is an essential substance for the synthesis of nucleotides in eukaryotic cells. Folate receptor 1 (FLOR1) is a glycoprotein that mediates the endocytosis of folic acid by cells. It is highly expressed on the surface of many malignant tumor cells, making it a new target that is widely studied in the field of tumor diagnosis and treatment. MMAE is a derivative of sea urchin toxin. Currently, a variety of antibody-drug conjugates (ADCs) with MMAE as the drug carrier have been approved for marketing by domestic and foreign regulatory agencies or are in the development stage. The treatment areas cover breast cancer, lymphoma, gynecological tumors and sedatives. Compound X, which dual-targets FLOR1 and PSMA, has shown significant anti-tumor activity and good safety in preclinical studies, supporting its entry into clinical development.

Although compound X showed good efficacy as a single treatment, not all patients have high expression of PSMA and FOLR1, which means that single treatment cannot achieve the desired effect for patients with low receptor expression. Therefore, it is necessary to explore better treatment options.

Summary of the invention

Based on this, the purpose of the present disclosure is to provide a drug combination for treating prostate cancer and its application, wherein the drug combination can synergistically enhance the efficacy when used to treat prostate cancer.

In order to achieve the above objectives, the present invention adopts the following technical solutions:

In a first aspect, the present disclosure provides a use of a drug combination in the preparation of a drug for treating prostate cancer, the drug combination comprising (a) compound X or a pharmaceutically acceptable salt thereof; and (b) an androgen receptor antagonist or a pharmaceutically acceptable salt thereof; wherein the structural formula of compound X is:

In a preferred embodiment, the structural formula of the compound X is:

In a preferred embodiment, the androgen receptor antagonist is enzalutamide.

In a preferred embodiment, the drug combination synergistically inhibits the proliferation of prostate cancer cells.

In a preferred embodiment, the drug combination synergistically promotes apoptosis of prostate cancer cells.

In some embodiments, the dosage form of the pharmaceutical combination includes tablets, granules, capsules, solutions, powders, injections or pills.

In a second aspect, the present disclosure further provides a drug combination for treating prostate cancer, the drug combination comprising (a) compound X or a pharmaceutically acceptable salt thereof; and (b) an androgen receptor antagonist or a pharmaceutically acceptable salt thereof; wherein the structural formula of compound X is:

In a preferred embodiment, the structural formula of the compound X is:

In a preferred embodiment, the androgen receptor antagonist is enzalutamide.

In a preferred embodiment, the compound X and the androgen receptor antagonist may be administered separately or simultaneously.

In a third aspect, the present disclosure further provides a drug for treating prostate cancer, the drug comprising a main active ingredient and a pharmaceutically acceptable excipient; the main active ingredient comprises the drug combination defined in the first aspect or the second aspect.

In a preferred embodiment, the auxiliary material includes at least one of a filler, a bulking agent, a binder, a humectant, a disintegrant, a solubilizing agent, an adsorbent, a diluent, a solubilizing agent, an emulsifier, a lubricant, a wetting agent, a suspending agent, a flavoring agent and a fragrance.

In a fourth aspect, the present disclosure also provides a method for preventing or treating prostate cancer, the method comprising administering a therapeutically effective amount of the drug combination defined in the first aspect or the second aspect or the drug defined in the third aspect to a patient in need thereof.

In a fifth aspect, the present disclosure further provides a kit of parts comprising a therapeutically effective amount of the drug combination defined in the first aspect or the second aspect or the drug defined in the third aspect.

The present disclosure has found through research that compound X and enzalutamide can synergistically enhance the efficacy when combined to treat prostate cancer, and can better inhibit the proliferation of prostate cancer cells and promote the apoptosis of prostate cancer cells. The present disclosure has verified the potential value of the combined treatment of compound X and enzalutamide through experiments, proving that the efficacy of the combined use of the two is far greater than that of the single drug, providing a better treatment plan for the treatment of prostate cancer, and has broad application prospects.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a result of detecting the survival rate of each group of LNCAP cells treated in Example 1.

FIG. 2 is the calculation result of the drug synergy index of each group of LNCAP cells treated in Example 1.

FIG. 3 shows the survival rate test results of each group of 22Rv1 cells treated in Example 2.

FIG. 4 is the calculation results of the drug synergy index for each group of 22Rv1 cells treated in Example 2.

Figure 5 shows the apoptosis detection results of LNCAP cells in the Control group, the Compound X 8 ng/μL single treatment group, the Enzalutamide 200 μM single treatment group, and the Compound X 8 ng/μL + Enzalutamide 200 μM combined treatment group in Example 3.

Figure 6 shows the apoptosis rate of LNCAP cells in the Control group, the Compound X 8 ng/μL single treatment group, the Enzalutamide 200 μM single treatment group, and the Compound X 8 ng/μL + Enzalutamide 200 μM combined treatment group in Example 3.

7 shows the apoptosis detection results of LNCAP cells in the Control group, the Compound X 4 ng/μL single treatment group, the Enzalutamide 100 μM single treatment group, and the Compound X 4 ng/μL + Enzalutamide 100 μM combined treatment group in Example 3.

FIG8 shows the 22Rv1 cells in the Control group, the Compound X 8 ng/μL single treatment group, the Enzalutamide 200 μM single treatment group, and the Compound X 8 ng/μL + Enzalutamide 200 μM combined treatment group in Example 4. The results of apoptosis detection.

Figure 9 shows the apoptosis rate of 22Rv1 cells in the Control group, the Compound X 8 ng/μL single treatment group, the Enzalutamide 200 μM single treatment group, and the Compound X 8 ng/μL + Enzalutamide 200 μM combined treatment group in Example 4.

Detailed ways

The experimental methods in the following examples of the present disclosure without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. The various commonly used chemical reagents used in the examples are all commercially available products.

Unless otherwise defined, all technical and scientific terms used in the present disclosure have the same meanings as those commonly understood by those skilled in the art to which the present disclosure belongs. The terms used in the specification of the present disclosure are only for the purpose of describing specific embodiments and are not intended to limit the present disclosure.

The term "comprising" or "including" of the present disclosure refers to including all elements, integers or steps, but does not exclude any other elements, integers or steps. In this article, when the term "comprising" or "including" is used, unless otherwise specified, the situation consisting of the elements, integers or steps mentioned is also covered.

The term "plurality" used in this disclosure refers to two or more than two. "And/or" describes the association relationship of associated objects, indicating that three relationships may exist. For example, A and/or B may represent the following three situations: A exists alone, A and B exist at the same time, and B exists alone. The character "/" generally indicates that the associated objects are in an "or" relationship.

The term "prevention" in this disclosure includes the inhibition or delay of the occurrence or frequency of a disease or disorder or its symptoms, and generally refers to the administration of a drug before the disorder or symptom occurs, particularly before the disorder or symptom occurs in an individual at risk.

The term "treatment" in this disclosure refers to the slowing, arresting or reversing of the progression of a disease such as cancer in a subject as evidenced by the alleviation or elimination of clinical or diagnostic symptoms of the disease. Treatment may include, for example, reducing the severity of symptoms, the number of symptoms or the frequency of recurrence, such as tumor growth inhibition, tumor growth retardation or regression of existing tumors.

The term "pharmaceutical combination" in the present disclosure refers to a non-fixed combination product or a fixed combination product, such as a kit. The term "non-fixed combination" means that the active ingredients (e.g., (a) Compound X or a pharmaceutically acceptable salt thereof, and (b) an androgen receptor antagonist or a pharmaceutically acceptable salt thereof) are administered to a patient simultaneously, without specific time restrictions, or at the same or different time intervals, sequentially as separate entities, wherein such administration provides preventive or therapeutically effective levels of the two active agents in the patient's body. The term "fixed combination" means that two active agents are administered to a patient simultaneously in the form of a single entity. Preferably, the dosage and/or time interval of the two active agents are selected so that the combined use of the parts can produce an effect greater than that achieved by using either alone when treating a disease or condition, and each component can be in a separate formulation, which can be the same or different.

The term "therapeutically effective amount" in the present disclosure refers to the amount of combined administration that, when administered in combination, induces the desired biological or medical response, i.e., inhibits or improves one or more cancers. For example, when referring to combined therapy, the term "therapeutically effective amount" as used herein is the amount of antibody and the amount of chemotherapeutic drug that produce a therapeutically effective and/or synergistic combined effect when administered together (sequentially or simultaneously) on the same day or different days in a treatment cycle. Moreover, those skilled in the art will recognize that in the case of combined therapy with a therapeutically effective amount (such as the above examples), the amount of the chemical drug alone may or may not be therapeutically effective.

The term "administer" in the present disclosure refers to the physical introduction of each active ingredient in the drug combination of the present disclosure into an individual using any of a variety of methods and delivery systems known to those skilled in the art. The administration routes of each active ingredient in the drug combination of the present disclosure include oral, intravenous (e.g., infusion (also known as drip) or injection), intramuscular, subcutaneous, intraperitoneal, spinal, local or other routes. Accordingly, each active ingredient in the drug combination of the present disclosure can be formulated into capsules, tablets, injections (including infusions or injections), syrups, sprays, lozenges, liposomes or suppositories, etc.

The term "pharmaceutically acceptable salt" in the present disclosure includes, but is not limited to, acid addition salts or base addition salts, such as acid addition salts formed by compound X with inorganic acids, such as hydrochloride, hydrobromide, carbonate, bicarbonate, phosphate, sulfate, sulfite, nitrate, etc.; and acid addition salts formed by compound X with organic acids, such as formate, acetate, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethanesulfonate, benzoate, salicylate, stearate and salts with the formula HOOC-(CH ₂ )n-COOH (wherein n The salts formed by the alkanedicarboxylic acid (0, 1, 2, 3 or 4) are also included. Also included are base addition salts formed by the compound X having an acidic group and a pharmaceutically acceptable cation such as sodium, potassium, calcium, aluminum, lithium and ammonium.

The chemical formula of the term "enzalutamide" in the present disclosure is 4-[3-[4-cyano-3-(trifluoromethyl)phenyl]-5,5-dimethyl-4-oxo-2-thione-1-imidazolidinyl]-2-fluoro-N-methylbenzamide. Enzalutamide is an androgen receptor antagonist that can competitively inhibit the binding of androgens to the receptor, and can inhibit the nuclear translocation of the androgen receptor and the interaction of the receptor with DNA.

The following is a description of the reagents used in the following examples: Enzalutamide was purchased from Selleck, item number S1250-5mg.

Example 1

This example studies the effect of the combined use of compound X and enzalutamide on the activity of prostate cancer cells LNCAP.

After routine culture of LNCAP cells, the cells were treated with drugs, and the following experimental groups were set up: the compound X treatment group had final concentrations of 9.6 ng/μL (recorded as 5 groups), 4.8 ng/μL (recorded as 4 groups), 2.4 ng/μL (recorded as 3 groups), 1.2 ng/μL (recorded as 2 groups), and 0.6 ng/μL (recorded as 1 group); the enzalutamide treatment group had final concentrations of 400 μM (recorded as 5 groups), 200 μM (recorded as 4 groups), 100 μM (recorded as 3 groups), 50 μM (recorded as 2 groups), and 2 5μM (recorded as group 1); compound X and enzalutamide combined treatment group, including the following concentrations: compound X 9.6ng/μL+enzalutamide 400μM (recorded as group 5), compound X 4.8ng/μL+enzalutamide 200μM (recorded as group 4), compound X 2.4ng/μL+enzalutamide 100μM (recorded as group 3), compound X 1.2ng/μL+enzalutamide 50μM (recorded as group 2), compound X 0.6ng/μL+enzalutamide 25μM (recorded as group 1).

After 48 h of drug treatment, cells in each group were collected and the OD value was detected using a CCK8 kit (APEXBIO; Catalog No.: K1018-500T) (the operation was strictly carried out in accordance with the instructions of the kit) to calculate the cell viability, and then the mixed drug analysis software CalcuSyn was used to calculate the drug synergy index (when the synergy index <1, it means that the drugs have a synergistic effect).

The results are shown in Figure 1-2. Compared with the compound X alone treatment group and the enzalutamide alone treatment group, the compound X and enzalutamide combined treatment group had a stronger inhibitory effect on LNCAP cell viability. The synergy index of compound X and enzalutamide was <1 when they were co-administered at different concentrations, indicating that they all showed synergistic effects. The two can work synergistically in inhibiting prostate cancer.

Example 2

This example studies the effect of the combined use of compound X and enzalutamide on the activity of prostate cancer cell 22Rv1.

After routine culture of 22Rv1 cells, the cells were treated with drugs, and the following experimental groups were set up: the compound X single treatment group, the final concentrations were 9.6 ng/μL (recorded as 5 groups), 4.8 ng/μL (recorded as 4 groups), 2.4 ng/μL (recorded as 3 groups), 1.2 ng/μL (recorded as 2 groups), and 0.6 ng/μL (recorded as 1 group); the enzalutamide single treatment group, the final concentrations were 400 μM (recorded as 5 groups), 200 μM (recorded as 4 groups), 100 μM (recorded as 3 groups), 50 μM (recorded as 2 groups), 2 5μM (recorded as group 1); compound X and enzalutamide combined treatment group, including the following concentrations: compound X 9.6ng/μL+enzalutamide 400μM (recorded as group 5), compound X 4.8ng/μL+enzalutamide 200μM (recorded as group 4), compound X 2.4ng/μL+enzalutamide 100μM (recorded as group 3), compound X 1.2ng/μL+enzalutamide 50μM (recorded as group 2), compound X 0.6ng/μL+enzalutamide 25μM (recorded as group 1).

After 48 h of drug treatment, cells in each group were collected, and the OD value was detected using the CCK8 kit to calculate the cell viability, and then the drug synergy index was calculated using the mixed drug analysis software CalcuSyn.

The results are shown in Figures 3-4. Compared with the compound X alone treatment group and the enzalutamide alone treatment group, the compound X and enzalutamide combined treatment group had a stronger inhibitory effect on the viability of 22Rv1 cells. The synergistic index of compound X and enzalutamide was <1 when they were co-administered at different concentrations, both showing a synergistic effect, indicating that the two can synergize in inhibiting prostate cancer.

Example 3

This example studies the effect of the combined use of compound X and enzalutamide on the apoptosis of prostate cancer cells LNCAP.

LNCAP cells were routinely cultured and then treated with drugs, and the following experimental groups were set up: compound X alone treatment group, the final concentrations were 8 ng/μL and 4 ng/μL, respectively; enzalutamide alone treatment group, the final concentrations were 200 μM and 100 μM, respectively; compound X and enzalutamide combined treatment group, including the following concentrations: compound X 8 ng/μL + enzalutamide 200 μM, compound X 4 ng/μL + enzalutamide 100 μM; Control group: treated with solvent DMSO, the volume concentration of DMSO in the culture system was 0.1%.

After 48 h of drug treatment, cells in each group were collected, and the apoptosis rate was detected using an apoptosis kit (GOONIE; catalog number: 100-101) combined with flow cytometry. The operation was carried out strictly in accordance with the instructions of the kit.

As shown in Figures 5-6, compared with the compound X 8ng/μL single treatment group and the enzalutamide 200μM single treatment group, the apoptosis rate of LNCAP cells in the compound X 8ng/μL + enzalutamide 200μM combined treatment group was significantly increased. As shown in Figure 7, compared with the compound X 4ng/μL single treatment group and the enzalutamide 100μM single treatment group, the apoptosis rate of LNCAP cells in the compound X 4ng/μL + enzalutamide 100μM combined treatment group was significantly increased. This shows that the two can synergistically enhance the effect in promoting the apoptosis of prostate cancer cells.

Example 4

This example studies the effect of the combined use of compound X and enzalutamide on the apoptosis of prostate cancer cell 22Rv1.

22Rv1 cells were routinely cultured and then treated with drugs. The following experimental groups were set up: compound X alone treatment group, with final concentrations of 8 ng/μL and 4 ng/μL, respectively; enzalutamide alone treatment group, with final concentrations of 200 μM and 100 μM, respectively; compound X and enzalutamide combined treatment group, including the following concentrations: compound X 8 ng/μL + enzalutamide 200 μM, compound X 4 ng/μL + enzalutamide 100 μM; Control group: treated with solvent DMSO, and the volume concentration of DMSO in the culture system was 0.1%.

After 48 h of drug treatment, cells in each group were collected, and the apoptosis rate was detected using an apoptosis kit (GOONIE; catalog number: 100-101) combined with flow cytometry. The operation was carried out strictly in accordance with the instructions of the kit.

As shown in Figures 8-9, compared with the compound X 8 ng/μL single treatment group and the enzalutamide 200 μM single treatment group, the apoptotic rate of 22Rv1 cells in the compound X 8 ng/μL + enzalutamide 200 μM combined treatment group was significantly increased, indicating that the two can synergistically enhance the effect in promoting the apoptosis of prostate cancer cells.

The above results indicate that compound X and enzalutamide can synergistically enhance the efficacy when combined to treat prostate cancer, and can better inhibit the proliferation of prostate cancer cells and promote the apoptosis of prostate cancer cells.

The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

The above-mentioned embodiments only express several implementation methods of the present disclosure, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the present disclosure. Those skilled in the art can make several modifications and improvements without departing from the concept of the present disclosure, which are all within the scope of protection of the present disclosure. Therefore, the scope of protection of the patent of the present disclosure shall be subject to the attached claims.

Claims (11)

1. A use of a drug combination in the preparation of a drug for treating prostate cancer, characterized in that the drug combination comprises (a) compound X or a pharmaceutically acceptable salt thereof; and (b) an androgen receptor antagonist or a pharmaceutically acceptable salt thereof; wherein the structural formula of compound X is:
   
2. The use according to claim 1, characterized in that the androgen receptor antagonist is enzalutamide.
3. The use according to claim 1 or 2, characterized in that the drug combination synergistically inhibits the proliferation of prostate cancer cells.
4. The use according to claim 1 or 2, characterized in that the drug combination synergistically promotes the apoptosis of prostate cancer cells.
5. The use according to claim 1 or 2, characterized in that the dosage form of the drug combination includes tablets, granules, capsules, solutions, powders, injections or pills.
6. A drug combination for treating prostate cancer, characterized in that the drug combination comprises (a) compound X or a pharmaceutically acceptable salt thereof; and (b) an androgen receptor antagonist or a pharmaceutically acceptable salt thereof; wherein the structural formula of compound X is:
   
7. The pharmaceutical combination according to claim 6, characterized in that the androgen receptor antagonist is enzalutamide.
8. According to the pharmaceutical combination of claim 6 or 7, the compound X and the androgen receptor antagonist can be administered separately or simultaneously.
9. A drug for treating prostate cancer, characterized in that the drug comprises a main active ingredient and a pharmaceutically acceptable excipient; the main active ingredient comprises the drug combination according to any one of claims 6 to 8.
10. The drug for treating prostate cancer as claimed in claim 9, characterized in that the excipients include at least one of a filler, a bulking agent, a binder, a humectant, a disintegrant, a solubilizing agent, an adsorbent, a diluent, a solubilizing agent, an emulsifier, a lubricant, a wetting agent, a suspending agent, a flavoring agent and a fragrance.
11. A complete set of medicine kits, comprising a therapeutically effective amount of the drug combination according to any one of claims 6 to 8 or the drug according to claim 9 or 10.