Method for treating cancer by combined application of Mobosertinib and anti-PD-1 antibody

Abstract

The invention discloses a method for treating cancers by combined application of Mobosertinib and an anti-PD-1 antibody, researches on the synergistic effect of Mobosertinib on non-EGFR exon 20 insertion mutation diseases and the synergistic effect of Mobosertinib and the anti-PD-1 antibody, and finds that the action mechanism of Mobosertinib is related to abnormal expression of SET. The invention provides a technical scheme of new application and combined application of Mobosertinib, and a new research direction is provided for treatment of cancers related to SET abnormal expression.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and particularly to a method for treating cancer by the combined application of Mobocertinib and an anti-PD-1 antibody. Background Art

[0002] The SET gene, whose full name is SET nuclear proto-oncogene, is located at position 9q34.11 of human chromosome. The protein encoded by this gene plays an important role in the nucleus, belongs to the INHAT complex and the SET complex, and is also a member of the Nucleosome assembly protein superfamily. SET is highly expressed in rapidly dividing cells and is an oncoprotein highly expressed in various types of tumors. It can interact with a variety of key factors to promote the tumor-related behaviors of cells. Research shows that the abnormal expression of SET can promote the occurrence and development of tumors.

[0003] Mobocertinib is a novel and highly selective small molecule tyrosine kinase inhibitor (TKI) targeting epidermal growth factor receptor (EGFR) and exon 20 insertion mutation (ex20ins) of human epidermal growth factor receptor 2 (HER2). In clinical trials, Mobocertinib also has significant therapeutic effects on adult patients with locally advanced or metastatic NSCLC with EGFR exon 20 insertion mutation whose disease progresses during or after platinum-containing chemotherapy. Since Mobocertinib is a targeted drug for EGFR exon 20 insertion mutation, there is no research on whether this drug has potential therapeutic effects in other diseases with non-EGFR exon 20 insertion mutations, nor is its molecular mechanism of action clear. In view of this, the present invention studies the effects of Mobocertinib on diseases with non-EGFR exon 20 insertion mutations, and provides a new application of Mobocertinib and a technical solution for combined application. Summary of the Invention

[0004] In view of the deficiencies of the prior art, the present invention provides a new application of Mobocertinib in diseases with non-EGFR exon 20 insertion mutations and its combined application with an anti-PD-1 antibody.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] The first aspect of the present invention provides a pharmaceutical composition for treating cancer, and the pharmaceutical composition comprises a first component and a second component.

[0007] The first component is Mobocertinib or a pharmaceutically acceptable salt thereof.

[0008] Mobocertinib is a kinase inhibitor targeting human epidermal growth factor receptor (EGFR). It is specifically used for the treatment of non-small cell lung cancer (NSCLC) caused by EGFR exon 20 insertion mutations, which are usually associated with a poor prognosis (compared with the "classical" EGFR mutations that cause NSCLC).

[0009] In the present invention, the pharmaceutically acceptable salt refers to a salt of an active compound (such as Mobocertinib) and is prepared by reacting the active compound with a suitable organic or inorganic acid or acid derivative. Pharmaceutically acceptable salts include but are not limited to hydrochloride, sulfate, phosphate, citrate, hydrobromide, acetate, benzoate, benzenesulfonate, tartrate, carbonate, citrate, gluconate, lactate, malate, mesylate, stearate, valerate, nitrate, sodium salt, calcium salt, potassium salt, zinc salt, and meglumine salt.

[0010] The second component is an anti-PD-1 antibody.

[0011] PD-1 (also known as CD279) is a type I transmembrane protein with a relative molecular weight of 55,000 - 60,000, mainly expressed on immune cells such as activated CD4+ T cells, CD8+ T cells, B cells, NK cells, monocytes, and dendritic cells, and promotes the maturation of T cells. Monoclonal antibodies against PD-1 can block the binding of PD-1 to its ligand, thereby restoring the tumor-killing function of T cells.

[0012] Furthermore, the antibody is a chimeric antibody, humanized antibody, or human antibody. Monoclonal antibodies derived from mammals are particularly preferred. Monoclonal antibodies derived from mammals include those produced by hybridomas and those produced by hosts transformed with an expression vector containing an antibody gene using genetic engineering methods.

[0013] Furthermore, the anti-PD-1 antibody is selected from anti-mouse PD-1 antibody, anti-human PD-1 antibody, and anti-cynomolgus monkey PD-1 antibody.

[0014] Furthermore, the anti-PD-1 antibody is an anti-mouse PD-1 antibody.

[0015] The composition of the present invention may further comprise a pharmaceutically acceptable carrier or excipient. The "pharmaceutically acceptable carrier or excipient" refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or any type of formulation aid. The carrier should be biologically acceptable, meaning that it is compatible with the other components in the formulation and harmless to the patient, and will not cause adverse reactions (such as an immune response) when administered to the host.

[0016] Furthermore, the pharmaceutical composition also includes pharmaceutically acceptable auxiliary materials, and the pharmaceutically acceptable auxiliary materials are selected from one or more of the following: diluent, excipient, filler, binder, wetting agent, disintegrant, emulsifier, solubilizing agent, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH regulator, antioxidant or bacteriostatic agent.

[0017] Furthermore, the dosage forms of the first component and the second component in the pharmaceutical composition are the same or different.

[0018] Furthermore, the dosage forms include drops, tablets, capsules, granules, films, gels, powders, emulsions, dripping pills, suppositories, aerosols, sprays, powder aerosols, patches, solutions, sterile powders for injection, ointments or creams.

[0019] Furthermore, the first component and the second component are administered simultaneously or successively. Specifically, the interval between successive administrations can be 0, 1, 2, 3, 4, 5, 6, 7 or more days.

[0020] Furthermore, the first component is administered daily, and the second component is administered every three days.

[0021] Furthermore, the dose ratio of the first component to the second component is 2 - 2.5:1.

[0022] Furthermore, the dose of the first component is 3 mg / kg, and the dose of the second component is 30 μg.

[0023] In the present invention, regarding the dose ratio of the first component and the second component, those skilled in the art can appropriately adjust and convert the dose ratio according to the species of the administration object, through differences in body surface area, pharmacological, physiological and anatomical factors, pharmacokinetic parameters, metabolic functions, receptors and lifespan, etc. For example, the dose ratio can be reasonably adjusted and converted through allometric scaling.

[0024] Furthermore, the cancer is a cancer with abnormal SET expression.

[0025] In the present invention, SET includes wild-type, mutant or fragments thereof. This term encompasses full-length, unprocessed SET, as well as any form of SET derived from processing in cells. This term encompasses naturally occurring variants of SET (such as splice variants or allelic variants). This term encompasses, for example, the SET gene, human SET, and SET from any other vertebrate source, including mammals such as primates and rodents (such as mice and rats). In a specific embodiment of the present invention, SET is a human gene with a gene ID of 6418.

[0026] Furthermore, the cancers with abnormal SET expression include solid tumors with abnormal SET expression and hematological tumors with abnormal SET expression.

[0027] Furthermore, the solid tumors with abnormal SET expression include colorectal cancer, melanoma, breast cancer, ovarian cancer, liver cancer, non-small cell lung cancer, and kidney cancer.

[0028] Furthermore, the hematological tumors with abnormal SET expression include leukemia and Hodgkin lymphoma.

[0029] Furthermore, the cancer with abnormal SET expression is colorectal cancer or melanoma.

[0030] In the present invention, the term "treatment" refers to the process of intervening or altering a specific health state, including eliminating the cause, symptomatic treatment or supportive treatment, and does not necessarily mean curing or completely eliminating the disease, condition or symptoms of the disease or condition. Treatment can refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction or improvement in the severity of the symptoms / effects of a determined disease.

[0031] The second aspect of the present invention provides the use of the pharmaceutical composition described in the first aspect of the present invention in the preparation of a medicament for treating cancer, wherein the cancer is a cancer with abnormal SET expression.

[0032] Furthermore, the cancers with abnormal SET expression include solid tumors with abnormal SET expression and hematological tumors with abnormal SET expression.

[0033] Furthermore, the solid tumors with abnormal SET expression include colorectal cancer, melanoma, breast cancer, ovarian cancer, liver cancer, non-small cell lung cancer, and kidney cancer.

[0034] Furthermore, the hematological tumors with abnormal SET expression include leukemia and Hodgkin lymphoma.

[0035] Furthermore, the cancer with abnormal SET expression is colorectal cancer or melanoma.

[0036] The third aspect of the present invention provides the use of Mobocertinib or a pharmaceutically acceptable salt thereof, and the use includes any one of the following:

[0037] 1) Use in the preparation of a drug for enhancing a patient's sensitivity to anti-PD-1 antibodies;

[0038] 2) Use in the preparation of a cancer treatment drug for non-EGFR exon 20 insertion mutations.

[0039] The administration subjects of the pharmaceutical composition of the present invention or Mobocertinib or a pharmaceutically acceptable salt thereof include mammals or tumor cells of the mammals. The mammals are preferably rodents, artiodactyls, perissodactyls, lagomorphs, primates, etc. The primates are preferably monkeys, apes or Homo sapiens. The subject can be a patient suffering from a tumor, or an ex vivo tumor cell of a patient suffering from a tumor.

[0040] The "drug", "pharmaceutical composition", "cancer drug", "drug for cancer treatment", "tumor drug" in the present invention have the same meaning, and they represent substances or combinations of substances that can treat cancer. The pharmaceutical composition provided by the present invention has the effect of treating cancer. The "treatment" includes preventing or delaying the appearance of symptoms and complications of diseases (such as cancer), and also includes prolonging the survival period of cancer patients, improving the quality of life, alleviating symptoms, shrinking or even eliminating tumors, curbing tumor metastasis, and preventing tumor recurrence. In the present invention, "cancer treatment" and "inhibiting cell proliferation" have the same meaning.

[0041] Furthermore, the patient is a cancer patient.

[0042] Furthermore, the cancer is a cancer with abnormal SET expression.

[0043] Furthermore, the cancer with abnormal SET expression includes solid tumors with abnormal SET expression and hematological tumors with abnormal SET expression.

[0044] Furthermore, the solid tumors with abnormal SET expression include colorectal cancer, melanoma, breast cancer, ovarian cancer, liver cancer, non-small cell lung cancer, renal cancer.

[0045] Furthermore, the hematological tumors with abnormal SET expression include leukemia, Hodgkin lymphoma.

[0046] Furthermore, the cancer with abnormal SET expression is colorectal cancer or melanoma.

[0047] Furthermore, the antibody is a chimeric antibody, a humanized antibody or a human antibody.

[0048] Further, the anti-PD-1 antibody is selected from anti-mouse PD-1 antibody, anti-human PD-1 antibody, and anti-cynomolgus monkey PD-1 antibody.

[0049] Further, the anti-PD-1 antibody is an anti-mouse PD-1 antibody.

[0050] Further, the cancer with non-EGFR exon 20 insertion mutation is a cancer with abnormal SET expression.

[0051] Advantages and beneficial effects of the present invention: The present invention provides a new application of Mobocertinib in the treatment of cancers with non-EGFR exon 20 insertion mutation, and the application of Mobocertinib in combination with an anti-PD-1 antibody in the treatment of cancers or in a drug for enhancing the sensitivity of patients to an anti-PD-1 antibody. Through the study of its mechanism of action, it provides a new research direction for the treatment of cancers related to abnormal SET expression. Description of the Drawings

[0052] Figure 1 It is a WB result diagram of the inhibitory effect of Mobocertinib on SET expression. Among them, A is the SET expression level diagram after treating colorectal cancer MC38 cells with gradient concentrations of Mobocertinib, B is the SET expression level diagram after treating colorectal cancer MC38 cells with MG132, CHQ, and Mobocertinib, C is the SET expression level diagram after treating melanoma B16 cells with gradient concentrations of Mobocertinib, and D is the SET expression level diagram after treating melanoma B16 cells with MG132, CHQ, and Mobocertinib.

[0053] Figure 2 It is a statistical chart of the tumor inhibition rate of the combined application of Mobocertinib and an anti-PD-1 antibody on the subcutaneous tumorigenesis model of colorectal cancer MC38 cells in mice.

[0054] Figure 3 It is a statistical chart of the tumor inhibition rate of the combined application of Mobocertinib and an anti-PD-1 antibody on the subcutaneous tumorigenesis model of melanoma B16 cells in mice.

[0055] Figure 4 It is a statistical chart of the tumor weight on the 19th day of the subcutaneous tumorigenesis model of melanoma B16 cells in mice with the combined application of Mobocertinib and an anti-PD-1 antibody. Detailed Embodiments

[0056] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative efforts belong to the scope of protection of the present invention.

[0057] Example

[0058] I. Experimental materials

[0059] 1. Mouse strain: C57 BL / 6, 6-week-old male and female mice with a body weight in the range of 20 - 25 g.

[0060] 2. Reagent catalog numbers: Mobocertinib (MedChemExpress, AP32788); anti-mouse PD-1 (BioXCell, BE0146).

[0061] II. Experimental methods and results

[0062] 1. Inhibitory effect of Mobocertinib on SET

[0063] Previous studies have shown that the small molecule compound Mobocertinib can regulate the expression of SET protein. Cells with non-EGFR exon 20 insertion mutations and high SET expression were used to further verify the inhibitory effect of Mobocertinib on SET expression. Taking colorectal cancer MC38 tumor cells and melanoma B16 tumor cells as cell models, Western Blot experiments were used to examine the expression of SET protein in the cell models treated with Mobocertinib. It was found that Mobocertinib could gradually reduce the expression of SET in tumor cells (the results are shown in Figure 1 A and C). In addition, by treating with the protein synthesis inhibitors MG132 and CHQ, it was found that MG132 treatment could partially reverse the decrease in SET expression caused by Mobocertinib (the results are shown in Figure 1 B and D), further indicating that Mobocertinib may degrade the expression of SET protein through the proteasome pathway. Therefore, taking colorectal cancer and melanoma with abnormal SET expression reported in the existing literature as examples, the study on whether Mobocertinib has a therapeutic effect on cancers with abnormal SET expression was carried out.

[0064] 2. Subcutaneous tumorigenesis model of colorectal cancer in mice

[0065] 1) Method: Subcutaneous tumor-bearing mice inoculated with mouse colorectal cancer tumor cells MC38 were divided into four groups for treatment: (1) control group; (2) single treatment with Mobocertinib at a dose of 3 mg / kg (daily, intraperitoneal injection); (3) single treatment with anti-PD-1 antibody at a dose of 30 μg (once every three days, intraperitoneal injection); (4) combined treatment with Mobocertinib at a dose of 3 mg / kg and anti-PD-1 antibody at a dose of 30 μg (Mobocertinib: daily, intraperitoneal injection; anti-PD-1 antibody: once every three days, intraperitoneal injection).

[0066] 2) Results: Around 25 days, we found that single treatment with Mobocertinib could inhibit the growth of colorectal cancer tumor cells without EGFR exon 20 insertion mutation, and the tumor inhibition rate was about 41%; single treatment with anti-PD-1 antibody could also inhibit the growth of tumor cells, and the tumor inhibition rate was about 49%; when combining Mobocertinib and anti-PD-1 antibody for treatment, tumor growth could be significantly inhibited, and the tumor inhibition rate was about 80%, and the results were as Figure 2 shown.

[0067] Calculate the q value when combining Mobocertinib and anti-PD-1 antibody using the King's formula commonly used in this field. A q value > 1 represents a synergistic effect between the two.

[0068] The inhibition rate of 3 mg / kg Mobocertinib was 41% (n = 8) (EA),

[0069] The inhibition rate of 30 μg anti-PD-1 antibody was 49% (n = 8) (EB),

[0070] The combined inhibition rate of the two was 80% (n = 8) (E(A + B)).

[0071] Substitute into the King's formula: q = E(A + B) / (EA + EB - EA * EB) = 80% / (41% + 49% - 41% * 49%) = 1.144 > 1.

[0072] Based on the above calculations, the q value > 1, indicating that Mobocertinib and anti-PD-1 antibody have a synergistic effect when combined, and the combined treatment of Mobocertinib and anti-PD-1 for colorectal cancer has a synergistic effect.

[0073] 3. Subcutaneous tumor-bearing model of melanoma mice

[0074] 1) Method: Subcutaneous tumor-bearing mice inoculated with mouse melanoma tumor cells B16 were divided into four groups for treatment: (1) Control group; (2) Monotherapy with Mobocertinib at a dose of 3 mg / kg (daily, intraperitoneal injection); (3) Monotherapy with anti-PD-1 antibody at a dose of 30 µg (once every three days, intraperitoneal injection); (4) Combination therapy with Mobocertinib at a dose of 3 mg / kg and anti-PD-1 antibody at a dose of 30 µg (Mobocertinib: daily, intraperitoneal injection; anti-PD-1 antibody: once every three days, intraperitoneal injection).

[0075] 2) Results: Around 19 days, we found that monotherapy with Mobocertinib could inhibit the growth of melanoma tumor cells without EGFR exon 20 insertion mutations, and the tumor inhibition rate was approximately 38%; monotherapy with anti-PD-1 antibody could also inhibit the growth of tumor cells, and the tumor inhibition rate was approximately 46%; when combining Mobocertinib and anti-PD-1 antibody for treatment, tumor growth could be significantly inhibited, and the tumor inhibition rate was approximately 80%, as shown in Figure 3 shown. Comparing the tumor weights of mice on the 19th day, the trend was consistent with the growth curve of mice, as shown in Figure 4 shown.

[0076] Calculate the q value when combining Mobocertinib and anti-PD-1 antibody using the King's formula commonly used in this field. A q value > 1 indicates a synergistic effect between the two.

[0077] The inhibition rate of 3 mg / kg Mobocertinib was 38% (n = 8) (EA),

[0078] The inhibition rate of 30 µg anti-PD-1 antibody was 46% (n = 8) (EB),

[0079] The combined inhibition rate of the two was 80% (n = 8) (E(A + B)).

[0080] Substitute into the King's formula: q = E(A + B) / (EA + EB - EA * EB) = 80% / (38% + 46% - 38% * 46%) = 1.203 > 1.

[0081] Based on the above calculations, the q value > 1, indicating that Mobocertinib and anti-PD-1 antibody have a synergistic effect when combined, and Mobocertinib and anti-PD-1 combined therapy for melanoma has a synergistic effect.

[0082] The description of the above embodiments is only for understanding the method and its core idea of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. A pharmaceutical composition for treating cancer, characterized in that: The pharmaceutical composition comprises a first component and a second component; The first component is Mobocertinib or a pharmaceutically acceptable salt thereof; The second component is an anti-PD-1 antibody; Preferably, the antibody is a chimeric antibody, a humanized antibody or a human antibody; Preferably, the anti-PD-1 antibody is selected from anti-mouse PD-1 antibody, anti-human PD-1 antibody and anti-cynomolgus monkey PD-1 antibody; Preferably, the anti-PD-1 antibody is an anti-mouse PD-1 antibody.

2. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition also includes pharmaceutically acceptable excipients, which are selected from one or more of the following: diluents, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH regulators, antioxidants or antibacterial agents.

3. The pharmaceutical composition according to claim 1, characterized in that The dosage forms of the first component and the second component in the pharmaceutical composition are the same or different; Preferably, the dosage form includes drops, tablets, capsules, granules, films, gels, powders, emulsions, pills, suppositories, aerosols, sprays, powder mists, patches, solutions, sterile powders for injection, ointments or creams.

4. The pharmaceutical composition according to claim 1, characterized in that The first component and the second component are administered simultaneously or sequentially, specifically, the interval between the sequential administrations may be 0, 1, 2, 3, 4, 5, 6, 7 or more days; Preferably, the first component is administered daily, and the second component is administered every three days; Preferably, the dosage ratio of the first component to the second component is 2-2.5:1; Preferably, the dosage of the first component is 3 mg / kg, and the dosage of the second component is 30 μg.

5. The pharmaceutical composition according to claim 1, characterized in that The cancer is a cancer in which SET is abnormally expressed; The cancers with abnormal SET expression include solid tumors with abnormal SET expression and blood tumors with abnormal SET expression; Preferably, the solid tumors with abnormal expression of SET include colorectal cancer, melanoma, breast cancer, ovarian cancer, liver cancer, non-small cell lung cancer, and renal cancer; Preferably, the blood tumors with abnormal expression of SET include leukemia and Hodgkin's lymphoma; Preferably, the cancer in which SET is abnormally expressed is colorectal cancer or melanoma.

6. Use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of a drug for treating cancer, characterized in that: The cancer is a cancer in which SET is abnormally expressed; The cancers with abnormal SET expression include solid tumors with abnormal SET expression and blood tumors with abnormal SET expression; Preferably, the solid tumors with abnormal expression of SET include colorectal cancer, melanoma, breast cancer, ovarian cancer, liver cancer, non-small cell lung cancer, and renal cancer; Preferably, the blood tumors with abnormal expression of SET include leukemia and Hodgkin's lymphoma; Preferably, the cancer in which SET is abnormally expressed is colorectal cancer or melanoma.

7. Use of Mobocertinib or a pharmaceutically acceptable salt thereof, characterized in that: The application includes any of the following: 1) Application in the preparation of drugs that enhance patients' sensitivity to anti-PD-1 antibodies; 2) Application in the preparation of therapeutic drugs for cancers with non-EGFR exon 20 insertion mutations.

8. The use according to claim 7, characterized in that: The patient is a cancer patient; Preferably, the cancer is a cancer in which SET is abnormally expressed; The cancers with abnormal SET expression include solid tumors with abnormal SET expression and blood tumors with abnormal SET expression; Preferably, the solid tumors with abnormal expression of SET include colorectal cancer, melanoma, breast cancer, ovarian cancer, liver cancer, non-small cell lung cancer, and renal cancer; Preferably, the blood tumors with abnormal expression of SET include leukemia and Hodgkin's lymphoma; Preferably, the cancer in which SET is abnormally expressed is colorectal cancer or melanoma.

9. The use according to claim 7, characterized in that: The antibody is a chimeric antibody, a humanized antibody or a human antibody; Preferably, the anti-PD-1 antibody is selected from anti-mouse PD-1 antibody, anti-human PD-1 antibody and anti-cynomolgus monkey PD-1 antibody; Preferably, the anti-PD-1 antibody is an anti-mouse PD-1 antibody.

10. The use according to claim 7, characterized in that: The cancer with non-EGFR exon 20 insertion mutation is a cancer with abnormal expression of SET.

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