Application of levetiracetam in preparation of medicine for reversing PD-1 inhibitor drug resistance or treating breast cancer

Abstract

The invention belongs to the technical field of drug research, and particularly relates to application of levetiracetam in preparation of drugs for reversing PD-1 inhibitor drug resistance or treating breast cancer. Tests prove that levetiracetam can remodel a tumor immune microenvironment and activate anti-tumor immunity to achieve the effect of treating breast cancer, reverse PD-1 inhibitor drug resistance, inhibit the growth of PD-1 inhibitor drug-resistant cancers and further improve the clinical remission rate of PD-1 inhibitor treatment.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical research, and specifically relates to an application of levetiracetam in the preparation of a drug for reversing PD-1 inhibitor resistance or treating breast cancer. Background Art

[0002] Resistance to PD-1 inhibitors is a major challenge in current cancer immunotherapy. Clinical remission rates are only 10%-30%, and some cancer types (such as triple-negative breast cancer, prostate cancer, and pancreatic cancer) are inherently insensitive to PD-1 inhibitors. The mechanisms of resistance are diverse and can be broadly categorized as primary and secondary. Primary resistance primarily involves abnormal basal expression of PD-1 and PD-L1, whose expression levels determine response to PD-1 inhibitors. A high tumor mutational burden also significantly influences response. Furthermore, mutations in key anti-tumor pathways, such as those in the IFN-γ and antigen presentation pathways, may contribute to primary resistance to PD-1 immune checkpoint inhibitors. Unlike primary resistance to PD-1 inhibitors, the mechanisms of secondary resistance remain incompletely understood and remain under investigation. Existing research suggests that inactivation of the IFN-γ response pathway and dysregulation of antigen presentation are potent inducing factors of acquired resistance. Furthermore, the loss of neoantigens may also contribute to resistance to PD-1 inhibitors. Currently, new strategies to reverse resistance to PD-1 inhibitors are a hot topic in tumor research.

[0003] Levetiracetam is a commonly used anti-epileptic drug. In clinical practice, it is mainly used to treat epilepsy, myoclonic seizures, and some neuropathic pain or bipolar disorder in adults and children as an adjuvant therapy. Unlike traditional anti-epileptic drugs, levetiracetam mainly acts through the following pathways: (1) Binding to the synaptic vesicle protein SV2A. SV2A is a glycoprotein on synaptic vesicles that participates in the regulation of neurotransmitter release. After binding to SV2A, levetiracetam may reduce the excessive release of excitatory transmitters such as glutamate from vesicles, thereby reducing abnormal neuronal discharges; (2) Regulating calcium ion channels, inhibiting N-type high-voltage calcium channels (N-type Ca²⁺ channels), reducing calcium ion influx into neurons, and stabilizing cell membrane excitability; (3) Affecting the GABAergic system, indirectly enhancing the inhibitory effect of GABA (inhibitory neurotransmitter), but unlike benzodiazepines, it does not directly act on GABA receptors; (4) Inhibiting excessive neuronal synchronization: blocking the spread of epileptiform discharges, especially effective for abnormal high-frequency discharges.

[0004] In recent years, reports have shown that levetiracetam has anti-tumor effects. Studies have shown that levetiracetam (LEV) can inhibit the progression of gliomas. LEV alone has been shown to inhibit glioma cell proliferation at the cellular level (PMID: 36281939). Levetiracetam also appears to have a sensitizing effect on existing anti-glioma treatments. Studies have shown that: (1) Levetiracetam enhances the inhibitory effect of temozolomide on glioma stem cell proliferation by reducing MGMT expression, promoting HDAC4 nuclear translocation and activating the apoptosis pathway (PMID 302143782018); (2) In a xenograft mouse model, levetiracetam can reduce the neuronal generation of miR-184-3p-enriched exosomes, inhibit the transformation of glioma stem cells from the anterior membrane to the mesenchyme, and increase the radiosensitivity of the tumor, thereby prolonging the survival time of tumor-bearing mice (PMID: 37963207); (3) Levetiracetam, IFN-α combined with temozolomide significantly prolonged the survival time of mice bearing orthotopic GSC-1 glioma and activated the apoptosis of U138 tumor cells (PMID: 33074386). In addition, levetiracetam also has certain anti-tumor effects in other tumor types. Studies have shown that in transgenic prostate adenocarcinoma mice that underwent surgical castration to simulate androgen deprivation therapy, levetiracetam reduced the incidence of high-grade prostatic intraepithelial neoplasia, adenocarcinoma and neuroendocrine tumors, thereby inhibiting tumor progression (PMID 33737929).

[0005] However, existing studies have not reported the therapeutic effect of levetiracetam on triple-negative breast cancer and PD-1 inhibitor-resistant cancer. Summary of the Invention

[0006] Based on this, the present invention provides a new use of levetiracetam, which can treat breast cancer and reverse PD-1 inhibitor resistance, providing a new and effective method for solving the problem of PD-1 inhibitor resistance in clinical practice.

[0007] The specific technical solutions provided by the present invention are as follows: The present invention provides an application of levetiracetam, wherein the application is any of the following: (1) Application in the preparation of drugs for treating breast cancer; (2) Application in the preparation of drugs for reversing PD-1 inhibitor resistance.

[0008] The present invention also provides a medicine for treating breast cancer, which uses the levetiracetam as the sole active ingredient.

[0009] As a preferred embodiment of the present invention, the breast cancer is triple-negative breast cancer.

[0010] As a preferred embodiment of the present invention, the drug is prepared by compounding levetiracetam with pharmaceutically acceptable excipients.

[0011] As a preferred embodiment of the present invention, the drug is an oral preparation.

[0012] The present invention also provides a drug for reversing drug resistance to PD-1 inhibitors, which contains the levetiracetam as the sole active ingredient.

[0013] As a preferred embodiment of the present invention, the drug is prepared by compounding levetiracetam with pharmaceutically acceptable excipients.

[0014] The present invention also provides a composition for treating tumors, which is prepared by compounding levetiracetam and a PD-1 inhibitor.

[0015] As a preferred embodiment of the present invention, the mass ratio of levetiracetam to PD-1 inhibitor is 10-15:1.

[0016] As a preferred embodiment of the present invention, the tumor is triple-negative breast cancer.

[0017] As a preferred embodiment of the present invention, the composition is prepared by compounding levetiracetam, a PD-1 inhibitor and pharmaceutically acceptable excipients.

[0018] As a preferred embodiment of the present invention, the composition is prepared as an oral preparation.

[0019] Furthermore, the oral preparation is a granule, tablet, capsule or liquid preparation.

[0020] The beneficial effects of the present invention are: The present invention discovered that levetiracetam can reshape the tumor immune microenvironment and activate anti-tumor immunity to achieve the therapeutic effect of breast cancer. It also reverses PD-1 inhibitor resistance, increases tumor sensitivity to PD-1 inhibitors, synergistically inhibits the progression of PD-1-resistant breast cancer, and further improves the clinical remission rate of PD-1 inhibitor-treated cancer. Because levetiracetam has a strong tumor immune activation effect, it provides a new strategy for the clinical preparation of anti-tumor drugs and the reversal of PD-1 inhibitor resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is a specific strategy of establishing a triple-negative breast cancer model in Balb / c mice using the 4T1 mouse breast cancer cell line and administering the drug intraperitoneally; Figure 2 This is a comparison of the actual tumors in each group; Figure 3 is the tumor volume growth curve of each group; Figure 4 is a bar graph of tumor weight in each group; Figure 5 is the content of myeloid-derived suppressor cells in the tumors of each group; Figure 6 CD8 + T cell content; Figure 7 is the weight growth curve of mice in each group; Figure 8 is the food intake curve of each group of mice; Figure 9 These are the HE staining images of the organs of mice in each group. DETAILED DESCRIPTION

[0022] The present invention will be further illustrated by the following specific examples, but they are not intended to limit the present invention.

[0023] Drug resistance to PD-1 inhibitors is one of the main challenges in current tumor immunotherapy, with a clinical remission rate of only 10% to 30%. In addition, some cancer types (such as triple-negative breast cancer, prostate cancer, and pancreatic cancer) are naturally insensitive to PD-1 inhibitors.

[0024] Levetiracetam is a commonly used anti-epileptic drug, mainly used in clinical practice for the treatment of epilepsy, myoclonic seizures, and some neuropathic pain or as an adjuvant therapy for bipolar disorder in adults and children. In recent years, the anti-tumor effects of levetiracetam have also been reported. Studies have found that levetiracetam can inhibit the progression of brain gliomas. In addition, levetiracetam also has certain anti-tumor effects in other tumor types. Studies have shown that in transgenic prostate adenocarcinoma mice that underwent surgical castration to simulate androgen deprivation therapy, levetiracetam reduced the frequency of high-grade prostatic intraepithelial neoplasia, adenocarcinoma, and neuroendocrine tumors, thereby inhibiting tumor progression. However, existing studies have not yet reported the therapeutic effects of levetiracetam on triple-negative breast cancer and PD-1 inhibitor-resistant cancers.

[0025] Based on this, the present invention provides the use of levetiracetam in the preparation of a drug for treating breast cancer or reversing PD-1 inhibitor resistance.

[0026] The present invention is confirmed by the following experiments: 0. Construction of triple-negative breast cancer model The mouse breast cancer cell line 4T1 was injected into the mammary fat pad of Balb / c mice. Over time, the 4T1 cells continued to proliferate, forming tumors in situ in the mammary glands of Balb / c mice, becoming a mouse triple-negative breast cancer model. This model is also a PD-1 inhibitor resistance model. The specific process is as follows Figure 1 shown.

[0027] 1. Grouping and Methods The model was randomly divided into four groups: the first group of mice was intraperitoneally injected with a drug solvent control, labeled as the PBS group; the second group of mice was intraperitoneally injected with levetiracetam 30 mg / kg / d for 14 consecutive days, labeled as the Lev group; the third group of mice was intraperitoneally injected with PD-1 inhibitor (BioXCell, BE0146) 200 ug / mouse every two days starting from the 7th day after the tumor grew out of the tumor, inoculated with 4T1 cells, and labeled as the Anti-PD-1 group; the fourth group of mice was given a combination of the second and third group's dosing methods, levetiracetam and PD-1 inhibitor, labeled as the Lev+Anti-PD-1 group. The mass ratio of levetiracetam to PD-1 inhibitor can vary between 10 and 15:1. The mass ratio selected in this example was 10:1. After 15 days, the tumors were uniformly collected, measured, photographed, and weighed for analysis.

[0028] During oral administration of the drug to the mouse model, the mice were weighed every two days for statistical analysis. Their food intake was also measured and analyzed. Flow cytometry was used to detect immune cells within the tumors, and the hearts, livers, spleens, lungs, and kidneys of the mice were collected for HE staining.

[0029] 3. Results Figures 2 to 4 The results showed that the tumor size and volume of the combination drug group (Lev+Anti-PD-1 group) were significantly smaller than those of the single drug drug group (Lev group and Anti-PD-1 group) and the control group, proving that levetiracetam can reverse PD-1 inhibitor resistance, and its combination with PD-1 inhibitors can synergistically inhibit the progression of PD-1 inhibitor-resistant triple-negative breast cancer.

[0030] Figure 5~Figure 6 It showed that compared with the control group and the single-drug group, the content of myeloid-derived immunosuppressive cells (immunosuppressive cells) in the tumor of the combination drug group was reduced, and the content of CD8-positive T cells (immune-activating cells) was increased.

[0031] Figure 7-Figure 8 The results showed that there was no difference in the body weight and food intake of mice in the combined drug group compared with the single drug group and the control group, proving that the combination of levetiracetam and PD-1 inhibitors has better drug safety.

[0032] Figure 9 The experimental results showed that there was no significant difference in organ weight and tissue morphology between the drug-treated group and the control group, further proving that the combination of levetiracetam and PD-1 inhibitors has good drug safety for the treatment of PD-1 inhibitor-resistant triple-negative breast cancer.

[0033] In summary, levetiracetam has a good effect against triple-negative breast cancer at a dose of 30 mg / kg / d in mice. This dose is converted to an equivalent dose of 2.44 mg / kg / d for humans according to the body surface area correction method. The total dose for a 60 kg adult is 122 mg per day, and the anti-epileptic dose is 1000 mg per day. Its anti-tumor dose is lower than the clinical anti-epileptic dose.

[0034] It should be noted that the implementation of the present invention has better feasibility and does not impose any form of limitation on the present invention. Any technician familiar with the field may use the technical content disclosed above to change or modify it into an equivalent effective embodiment. However, any modification or equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A use of levetiracetam, characterized in that, The application is any of the following: (1) Application in the preparation of drugs for treating breast cancer; (2) Application in the preparation of drugs for reversing PD-1 inhibitor resistance.

2. A drug for treating breast cancer, characterized in that: The invention uses the levetiracetam described in claim 1 as the sole active ingredient.

3. The drug according to claim 2, characterized in that The medicine is prepared by compounding levetiracetam with pharmaceutically acceptable excipients.

4. The drug according to claim 3, characterized in that The medicine is an oral preparation.

5. A drug for reversing drug resistance to PD-1 inhibitors, characterized in that: The invention contains the levetiracetam as claimed in claim 1 as the sole active ingredient.

6. The drug according to claim 5, characterized in that The medicine is prepared by compounding levetiracetam with pharmaceutically acceptable excipients.

7. A pharmaceutical composition for treating tumors, characterized in that: It is prepared by compounding the levetiracetam described in claim 1 with a PD-1 inhibitor.

8. The pharmaceutical composition according to claim 7, characterized in that The mass ratio of levetiracetam to PD-1 inhibitor is 10-15:

1.

9. The pharmaceutical composition according to claim 7, characterized in that The tumor is breast cancer.

10. The pharmaceutical composition according to claim 7, characterized in that The pharmaceutical composition is prepared by compounding levetiracetam, a PD-1 inhibitor and pharmaceutically acceptable excipients.

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