Application of naphthamide compound in treatment of meningioma

Abstract

The invention relates to an application of a naphthamide compound (I) or a pharmaceutically acceptable salt thereof in preparation of a medicine for treating meningioma, and provides a corresponding treatment method, and the structure of the compound (I) is shown in the specification. The compound (I) has an excellent inhibition effect on meningioma, can effectively treat meningioma, and has good safety and tolerance. # imgabs0 #

Description

Use of a naphthamide compound in treating meningioma Technical Field

[0001] The present application belongs to the field of medicine, and specifically relates to the use of a naphthamide compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating meningioma. Background Art

[0002] Meningiomas are the most common primary intracranial tumors, accounting for approximately two-fifths of all primary intracranial tumors. Depending on their location, meningiomas are categorized as intracranial meningiomas and ectopic meningiomas. Intracranial meningiomas form from cells in the arachnoid membrane within the skull, while ectopic meningiomas arise in tissues and organs without a meningeal covering, primarily evolving from residual arachnoid tissue from the embryonic period. The annual incidence of meningiomas is approximately 7.6 per 100,000 people. Patients are typically young at initial diagnosis, with 77.7% of patients aged 20-44 years.

[0003] Meningiomas can be divided into the following categories based on their location: convex meningiomas, parafalcine meningiomas, olfactory groove meningiomas, tuberculum sellae meningiomas, sphenoid ridge meningiomas, parasagittal meningiomas, middle skull base meningiomas, clival meningiomas, and foramen magnum meningiomas. Based on their clinical and pathological characteristics, the World Health Organization (WHO) classifies meningiomas into three grades: benign meningiomas (WHO grade I), atypical meningiomas (WHO grade II), and malignant meningiomas (WHO grade III). Benign meningiomas (WHO grade I) account for approximately 65% ​​to 80% of cases and are divided into the following nine subtypes: 1: Meningothelial, 2: Fibroblastic, 3: Transitional, 4: Psammomatous, 5: Angiomatous, 6: Microcystic, 7: Secretory, 8: Lymphoplasmacyte-rich, and 9: Metaplastic. Most meningiomas grow slowly and rarely recur after surgery. Atypical meningiomas (WHO grade II) account for approximately 20% to 35% of cases and are divided into the following three subtypes: 1: Atypical, 2: Clear cell, and 3: Chordoid. Malignant meningiomas (WHO grade III) account for approximately 3% of cases and are divided into three subtypes: 1. Rhabdoid, 2. Papillary, and 3. Anaplastic. WHO grades II-III are high-grade meningiomas characterized by high invasiveness, poor differentiation, and high recurrence and metastasis.

[0004] Meningiomas may be associated with neurofibromatosis, leading to complications such as visual impairment, visual field, olfactory or auditory disturbances, and limb movement disorders. Vision loss is related to the primary tumor site, with symptoms often occurring early and severe in the optic canal. Optic disc edema may develop early, and over time, it may progress to secondary optic atrophy and tubular meningioma. Early visual field constriction, particularly in cases involving the orbital apex, may result in the appearance of scotomas within the visual field.

[0005] The overall 10-year survival rate for meningiomas is 57.1%. The 5-year recurrence rate for WHO grade II meningiomas is approximately 50%, and the 10-year survival rate for recurrent patients is 53%. The 5-year recurrence rate for WHO grade III meningiomas is approximately 90%, and the 10-year survival rate for recurrent patients is 0%.

[0006] Meningiomas are usually diagnosed by brain MRI, and patients are stratified based on the presence or absence of symptoms and tumor size. Treatment is usually recommended based on tumor grade.

[0007] Patients with asymptomatic small tumors (≤3cm) can be observed regularly. In other cases, surgical resection should be performed as much as possible. If the patient is not eligible for surgery, radiotherapy can be chosen. Patients with grade I meningiomas can only undergo regular observation (conservative treatment) after surgery. If symptoms occur, postoperative radiotherapy can also be considered. For patients with grade II meningiomas that cannot be completely removed, postoperative radiotherapy is recommended. Certain patients (for example, patients who are not suitable for radiotherapy) can be observed regularly. Patients with grade II meningiomas who have undergone complete surgical resection can consider postoperative radiotherapy. All patients with surgically resected grade III meningiomas should receive adjuvant radiotherapy to enhance local control.

[0008] Systemic therapy is often considered for meningiomas that recur after surgery and / or radiotherapy or are not suitable for surgery or radiotherapy, but there is no recognized standard of care. Large randomized trials are lacking in this therapeutic area. ORRs for systemic treatment of meningiomas are generally very low. In a review of the outcomes of medical treatments for meningiomas by the Response Assessment in Neuro-Oncology (RANO) Working Group, 37 of 42 publications reported radiographic efficacy data. Among the 555 patients included in these 37 reports, the best radiographic response was reported as 1 CR (0.2%), 10 PR (1.8%), 13 MR (2.3%), and 343 SD (62%). The combined response rate of CR+PR+MR was 4.3%, indicating that tumor shrinkage is rarely observed. PFS-6 (6-month PFS rate) or ORR is commonly used as clinical trial endpoints in this therapeutic area. The RANO Working Group evaluated historical data from clinical trials involving medical treatments for meningioma. The weighted PFS6 for WHO grade I meningiomas was 29% (95% CI: 20.3%-37.7%). For WHO grade II / III meningiomas, the weighted mean PFS6 was 26% (95% CI: 19.3%-32.7%). In addition, in several small clinical trials, more than 20% or even 60% of patients experienced grade 3 or higher toxicity or discontinued treatment due to drug toxicity.

[0009] Therefore, there is an urgent need to find a safe and effective drug for the treatment of meningioma to solve the problem that there are currently no drugs available clinically.

[0010] Compound (I) was first disclosed in CN104860885A, and its structural formula is shown in the following formula (I):

[0011] Compound (I) is a VEGFR / CSF1R dual-target inhibitor with excellent activity, but its therapeutic effect on meningioma has not been studied.

[0012] Summary of the Invention

[0013] After long and in-depth research, the inventors of the present application unexpectedly discovered that compound (I) has excellent inhibitory activity against meningioma cells. Preliminary clinical studies have shown that compound (I) can effectively treat meningioma and has good safety and tolerability, and is expected to be developed into an anti-meningioma drug.

[0014] In a first aspect of the present application, there is provided a use of a compound (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating meningioma, wherein the compound (I) has the following structure:

[0015] In a second aspect of the present application, a pharmaceutical composition for treating meningioma is provided, comprising a therapeutically effective amount of compound (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, wherein the compound (I) has the following structure:

[0016] In a third aspect of the present application, a method for treating meningioma in an individual is provided, comprising:

[0017] A therapeutically effective amount of compound (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing compound (I) or a pharmaceutically acceptable salt thereof, is administered to an individual suffering from meningioma, wherein the compound (I) has the following structure:

[0018] In a fourth aspect of the present application, a compound (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound (I) or a pharmaceutically acceptable salt thereof, is provided for treating meningioma, wherein the compound (I) has the following structure:

[0019] In some embodiments of the present application, the meningiomas described in the first to fourth aspects above include but are not limited to the following meningiomas classified according to clinical pathological characteristics: WHO grade I meningioma, WHO grade II meningioma, and WHO grade III meningioma.

[0020] In some embodiments, the WHO grade I meningioma is a benign meningioma, including but not limited to: meningothelial meningioma, fibrous meningioma, transitional meningioma, psammoma meningioma, angiomatous meningioma, microcystic meningioma, secretory meningioma, lymphoplasmacytic meningioma, metaplastic meningioma. In a specific embodiment, the meningioma is selected from one or more of meningothelial meningioma, fibrous meningioma, transitional meningioma, psammoma meningioma, angiomatous meningioma, microcystic meningioma, secretory meningioma, lymphoplasmacytic meningioma, and metaplastic meningioma.

[0021] In some embodiments, the WHO grade II meningioma is an atypical meningioma, including but not limited to atypical meningioma, clear cell meningioma, and chordoid meningioma. In specific embodiments, the meningioma is selected from one or more of atypical meningioma, clear cell meningioma, and chordoid meningioma.

[0022] In some embodiments, the WHO grade III meningioma is a malignant meningioma, including but not limited to: rhabdoid meningioma, papillary meningioma, and anaplastic meningioma. In a specific embodiment, the meningioma is selected from one or more of rhabdoid meningioma, papillary meningioma, and anaplastic meningioma.

[0023] In some embodiments of the present application, the meningioma described in the first to fourth aspects above includes the following meningiomas classified by location of occurrence: intracranial meningioma and ectopic meningioma.

[0024] In some embodiments of the present application, the meningiomas described in the first to fourth aspects above include but are not limited to the following meningiomas classified according to the location of growth: convex meningioma, parafalcine meningioma, olfactory groove meningioma, tuberculum sellae meningioma, sphenoid ridge meningioma, parasagittal meningioma, middle skull base meningioma, clival meningioma, and foramen magnum meningioma.

[0025] In some embodiments of the present application, the meningioma described in the first to fourth aspects above is a meningioma that has failed or recurred after other treatments (also called recurrent or refractory meningioma).

[0026] In some embodiments, the other treatments include, but are not limited to, surgery, radiotherapy, and anti-tumor drug therapy.

[0027] In some embodiments, the anti-tumor drugs include but are not limited to chemotherapy drugs, targeted therapy drugs, immunotherapy drugs, etc., such as carrelizumab, bevacizumab, sunitinib, everolimus, etoposide, temozolomide, sirolimus, and honokiol. In a specific embodiment, the anti-tumor drugs are selected from one or more of carrelizumab, bevacizumab, sunitinib, everolimus, etoposide, temozolomide, sirolimus, and honokiol.

[0028] In some embodiments of the present application, the pharmaceutical composition described in the second to fourth aspects above is formulated into a clinically acceptable pharmaceutical dosage form, such as an oral preparation; preferably an oral solution.

[0029] In some embodiments of the present application, the therapeutically effective amount described in the third aspect is: a daily dosage of compound (I) of 5 mg to 30 mg, preferably 10 mg to 25 mg, more preferably 10 mg to 20 mg or 15 to 25 mg. In specific embodiments, the daily dosage of compound (I) is 10 mg, 15 mg, 20 mg or 25 mg.

[0030] In some embodiments of the present application, the compound (I) described in the first to fourth aspects above is administered as a single dose once daily, or is administered multiple times a day, or is used at intervals. In a preferred embodiment, the compound (I) is administered as a single dose once daily. In some embodiments of the present application, the compound (I) described in the first to fourth aspects above is administered for 21 consecutive days, optionally, multiple cycles of administration may be repeated.

[0031] In some embodiments of the present application, the individual described in the third aspect is a mammal, including but not limited to primates, cattle, horses, pigs, sheep, goats, dogs, cats, and rodents such as rats and mice. In a preferred embodiment, the individual is a human.

[0032] It should be understood that within the scope of this application, the above-mentioned technical features of this application and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one.

[0033] As used herein, the term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention formed with an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts.

[0034] "Pharmaceutically acceptable carrier" refers to a carrier that does not interfere with the biological activity of the active ingredient, including those commonly used in the pharmaceutical art, such as one or more compatible solid or liquid fillers or gel substances, which are suitable for mammalian, particularly human, administration and must be of sufficient purity and sufficiently low toxicity. "Compatibility" as used herein refers to the ability of the components of the composition to be compatible with the compounds of the present application, and with each other, without significantly reducing the efficacy of the compounds.

[0035] There is no particular limitation on the administration of the pharmaceutical composition of the present application. In some embodiments, representative administration routes include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.

[0036] Accordingly, the drug of the present application can be prepared into various clinically acceptable dosage forms. In some embodiments, the above dosage forms include oral dosage forms, injection dosage forms, local administration dosage forms or external dosage forms, etc.; preferably, an oral dosage form, such as a solid dosage form or an oral liquid, more preferably an oral solution.

[0037] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound from such compositions can be delayed in a certain portion of the digestive tract. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0038] Oral liquid dosage forms may include solutions, syrups, emulsions, suspensions, etc., preferably emulsions, more preferably self-emulsifying preparations. The self-emulsifying agent excipients include an oil phase, an emulsifier, an emulsifier co-emulsifier, and may also include an antioxidant. The oil phase includes medium-chain triglycerides (MCT), monocaprylin, dicaprylin, monoolein, monolinolein, corn oil, castor oil, sesame oil, peanut oil, olive oil, etc. The emulsifier includes polyoxyethylene (40) hydrogenated castor oil (RH40), polyoxyethylene (35) castor oil (Cremophor EL), vitamin E succinate polyethylene glycol ester (TPGS 1000), 15-hydroxystearic acid polyethylene glycol ester (HS15), linoleoyl polyoxyethylene (6) glyceride, linoleoyl polyoxyethylene (6) glyceride, caprylic acid capric acid polyethylene glycol glyceride (Labrasol ALF), etc. The co-emulsifier is selected from polyethylene glycol (PEG), diethylene glycol monoethyl ether (Transcutol HP), propylene glycol, glycerol, and the like. The antioxidant is selected from butylated hydroxytoluene, butylated hydroxyanisole, vitamin E, and propyl gallate. The self-emulsifying formulation can also be prepared as a capsule or tablet. Examples of oral solutions are described in the detailed description of the present application.

[0039] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions.

[0040] The dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier (including any preservatives, buffers, or propellants that may be required if necessary).

[0041] The compounds of the present application can be administered alone or in combination with other pharmaceutically acceptable anti-tumor drugs (such as chemotherapy drugs, targeted therapy drugs, immunotherapy drugs) or other treatment methods (such as radiotherapy, surgery).

[0042] The therapeutically effective amount described in this application refers to the effective dosage considered pharmaceutically, that is, the amount of active compound sufficient to significantly improve the condition without causing serious side effects. The specific dosage and frequency of administration should take into account factors such as the route of administration and the patient's health status, which can be determined by a skilled physician based on routine skills. In the first human dose escalation study (Phase I study), the most sensitive animal species is selected based on preclinical toxicology studies to estimate the maximum recommended starting dose for clinical trials. The starting dose is calculated according to the calculation method recommended by the "Technical Guidelines for Non-clinical Studies of Anti-tumor Drugs" and other guidelines. In some embodiments, in the Phase I clinical trial of Compound (I), in order to ensure the safety of the subjects, combined with the operability of the clinical trial and the specifications of the preparation, the Phase I clinical starting dose of Compound (I) is designed to be 5 mg. According to the results of the long-term toxicity test in animals, the maximum tolerated dose for humans is converted to 200 mg based on a human body weight of 60 kg. In the Phase I clinical trial of Compound (I), the safety, tolerability and efficacy of a continuous dosing regimen of 5 mg to 200 mg per day are studied. In some embodiments, for a person weighing 60 kg, the daily dosage of compound (I) is preferably 5 to 30 mg, more preferably 10 to 25 mg, more preferably 10 to 20 mg or 15-25 mg, and further preferably 10 mg, 15 mg, 20 mg or 25 mg. It can be administered as a single dose once daily, can be administered multiple times a day, or can be used at intervals, preferably as a single dose once daily. Preferably, the administration cycle is 21 consecutive days, and optionally, multiple cycles of administration can be repeated.

[0043] Throughout the specification and claims of this application, the words “comprises,” “includes,” and “contains” mean “including but not limited to,” and are not intended to exclude other parts, additives, components, or steps.

[0044] In some embodiments, compound (I) of the present application has an excellent inhibitory effect on meningioma. In a specific embodiment, compound (I) of the present application can significantly inhibit the viability, proliferation, migration or invasion of meningioma cells such as meningioma cell lines 10MM-Lee cells and CH157-MN cells. In another specific embodiment, compound (I) of the present application has a significant promoting effect on apoptosis of meningioma cells such as meningioma cell lines 10MM-Lee cells and CH157-MN cells.

[0045] In some embodiments, the compound (I) of the present application has been shown to be effective in treating meningioma in preliminary clinical studies. In some embodiments, the compound (I) of the present application has been shown to have good safety and tolerability in preliminary clinical studies.

[0046] Therefore, compared with the prior art, the present invention has one or more of the following main advantages:

[0047] (1) Compound (I) has excellent inhibitory activity against meningioma, significantly inhibiting the viability, proliferation, migration, and invasion of meningioma cells, such as meningioma cell lines IOMM-Lee cells and CH157-MN cells, and significantly promoting apoptosis of meningioma cells;

[0048] (2) Preliminary clinical studies have shown that compound (I) can effectively treat meningioma and has good safety and tolerability, and is expected to be developed as an anti-meningioma drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] FIG1 shows the inhibitory effect of compound (I) on the viability of human meningioma cells.

[0050] Figure 2 shows the inhibitory effect of compound (I) on the proliferation of human meningioma cells.

[0051] Figure 3 shows the inhibitory effect of compound (I) on the migration of human meningioma cells: Figure 3-1 and Figure 3-2 show the inhibitory effect of compound (I) on the migration of human meningioma cells IOMM-Lee, and Figure 3-3 shows the inhibitory effect of compound (I) on the migration of human meningioma cells CH157-MN; Figure A in Figure 3-2 and Figure 3-3 is a crystal violet staining image of human meningioma cells, and Figure B is a t-test analysis graph (*** indicates P < 0.001 compared with the blank control group).

[0052] Figure 4 shows the inhibitory effect of compound (I) on the invasion of human meningioma cells: Figure 4-1 and Figure 4-2 show the inhibitory effect of compound (I) on the invasion of human meningioma cells IOMM-Lee, and Figure 4-3 shows the inhibitory effect of compound (I) on the invasion of human meningioma cells CH157-MN; Figure A in Figure 4-1, Figure 4-2 and Figure 4-3 is a crystal violet staining image of human meningioma cells, and Figure B is a t-test analysis graph (*** indicates P < 0.001 compared with the blank control group).

[0053] Figure 5 shows the promoting effect of compound (I) on apoptosis of human meningioma cells: Figure 5-1 and Figure 5-2 show the promoting effect of compound (I) on apoptosis of human meningioma cells IOMM-Lee, and Figure 5-3 shows the promoting effect of compound (I) on apoptosis of human meningioma cells CH157-MN; Figure A in Figure 5-1, Figure 5-2 and Figure 5-3 is a flow cytometry graph, and Figure B is a t-test analysis graph (*** indicates P < 0.001 compared with the blank control group). DETAILED DESCRIPTION

[0054] The present application will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present application only and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are generally performed under conventional conditions or under conditions recommended by the manufacturer. Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to professionals in the field. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present application method. The preferred implementation methods and materials shown in the text are for demonstration purposes only.

[0055] Source or preparation of experimental materials:

[0056] Compound (I) was prepared by Shanghai Runshi Pharmaceutical Technology Co., Ltd.

[0057] The positive control compounds, reagents, and raw materials used in the experiments were all purchased commercially or prepared by ourselves.

[0058] Preparation method of in vitro test compound (I): After weighing, add DMSO to dissolve it into 10 mM, and dilute it with PBS to the required concentration before use.

[0059] The clinical trial test compound (I) preparation was prepared according to the following formula:

[0060] Specifications: 20ml: 200mg / bottle:

[0061] Preparation method:

[0062] According to the above prescription, polyoxyethylene (40) hydrogenated castor oil is melted in warm water at about 60°C in advance to a liquid state, medium-chain triglycerides, polyoxyethylene (40) hydrogenated castor oil, polyethylene glycol 400, and butylated hydroxytoluene are added to a liquid preparation tank, stirred evenly at 40-70°C, compound (I) is added, heated and stirred until the raw materials are completely dissolved, and then dispensed into oral solution bottles according to specifications.

[0063] Example 1: Trypan blue viable cell counting assay for the effect of compound (I) on the cell viability of human meningioma cell lines

[0064] 1. Experimental Methods

[0065] Human meningioma cell line IOMM-Lee cells (purchased from ATCC) were cultured in DMED medium containing 10% FBS and 100 U / mL penicillin-streptomycin in a 37°C, 5% CO2 incubator. Cells in the logarithmic growth phase were plated at 2×10 5Cells were evenly seeded at a density of 100 μg / ml in 6-well plates. After 24 hours of culture, the cells were divided into a blank control group and groups treated with different concentrations of Compound (I). The concentrations of Compound (I) were 0.4 μM, 0.8 μM, 1.6 μM, 2 μM, 10 μM, 20 μM, 60 μM, and 120 μM, with 3-4 replicate wells per group.

[0066] After culturing each cell group for 24 hours, the cells were digested with 0.25% trypsin. 50 μL of the cell suspension was mixed with 50 μL of 0.4% trypan blue solution to a final concentration of 0.2%. Then, 10 μL of the mixed suspension was added to a counting plate, and the cell count (total and dead cells) was counted under a microscope for statistical analysis.

[0067] 2. Experimental Results

[0068] The experimental results showed that, compared with the blank control group, compound (I) at different concentrations inhibited IOMM-Lee cell viability after 24 hours of treatment, and cell viability decreased with increasing concentration of compound (I). The results are shown in Table 1 and Figure 1. These results indicate that compound (I) can inhibit the cell viability of meningioma cells IOMM-Lee.

[0069] Table 1 Inhibitory effect of compound (I) on the viability of human meningioma cells

[0070] Note: Cell viability = (total number of cells - number of dead cells) / total number of cells × 100%

[0071] Example 2: Effect of EdU Detection Compound (I) on Proliferation of Human Meningioma Cell Lines

[0072] 1. Experimental Methods

[0073] The IOMM-Lee single cell suspension in the logarithmic growth phase obtained in Example 1 was inoculated into a 6-well plate at a cell density of 3×10 5 / ml, after overnight culture, they were divided into a blank control group and a compound (I) treatment group (concentration of 1μM, treatment for 24h). Then, each group was incubated with EdU (10μM) for 2h, the culture medium was discarded, and the cells were washed with PBS (PBS was used in the washing process in this experiment). Fix with 4% paraformaldehyde, discard the fixative and wash. Permeabilize with 0.3% Triton X100 (Triton X100), discard the permeabilization solution and wash. Then add the prepared click reaction solution (Click reaction solution, commercially available C0081 BeyoClick TMIncubate with an EdU-647 cell proliferation assay kit (EdU-647 Cell Proliferation Assay Kit) at room temperature in the dark for 30 minutes, discard the reaction solution, and wash. For nuclei staining, add 1 ml of 1× Hoechst 33342 solution to each well. Incubate at room temperature in the dark for 10 minutes. Aspirate the 1× Hoechst 33342 solution, and wash three times. Fluorescence detection and recording are then performed.

[0074] 2. Experimental Results

[0075] The results are shown in Figure 2. In the fluorescence detection photograph, the blue fluorescence channel shows Hochest 33342 staining of the cell nuclei, while the red fluorescence channel shows EdU staining of the proliferating cell nuclei. The sum of the two channels gives a magenta color, clearly indicating the ratio of proliferating cells to the total cell population. The experimental results show that compared with the blank control group, the number of EdU-labeled (proliferating) cells in the meningioma 10MM-Lee cells treated with compound (I) was significantly reduced. These results demonstrate that compound (I) can inhibit the proliferation of meningioma 10MM-Lee cells.

[0076] Example 3: Cell scratch assay to detect the effect of compound (I) on the migration of human meningioma cell lines

[0077] 1. Experimental Methods

[0078] (1) Inoculate a single IOMM-Lee cell suspension in the logarithmic growth phase into a 6-well plate at a cell density of 6×10 5 Each well was cultured for 24 hours, ensuring that cells grew and covered the entire cell culture dish. A 10 μL sterile pipette tip was used to scratch the cells perpendicular to the dish, and the original culture medium was discarded. The cells were washed three times with PBS to absorb the detached cells. Serum-free DMEM medium was used to culture the cells. The cells were divided into a blank control group and groups treated with different concentrations of Compound (I), with concentrations of 0.5 μM, 1 μM, and 2 μM, respectively. The cells were observed and recorded under a bright field microscope at 0 and 24 hours.

[0079] (2) Referring to the experimental method (1), the concentrations of the compound (I) treatment group were changed to 2 μM, 5 μM and 10 μM, respectively, and the human meningioma cell lines used were IOMM-Lee or CH157-MN.

[0080] 2. Experimental Results

[0081] The experimental results showed that after 24 hours of treatment with different concentrations of Compound (I) in IOMM-Lee or CH157-MN cells, the wound healing rate in the Compound (I)-treated group was significantly lower than that in the blank control group. Furthermore, with increasing concentrations of Compound (I), the wound healing rate gradually decreased, as shown in Figure 3 (Figures 3-1, 3-2, and 3-3). These results indicate that Compound (I) has a significant inhibitory effect on the migration of meningioma cells IOMM-Lee and CH157-MN in a dose-dependent manner.

[0082] Example 4: Transwell chamber assay for the effect of compound (I) on cell invasion of human meningioma cell lines

[0083] 1. Experimental Methods

[0084] (1) Coat the upper chamber of the pre-cooled Transwell chamber with 50 μL of matrigel and incubate the chamber coated with matrigel in a 37°C incubator for 6-8 hours to allow the matrigel to polymerize into a gel. Add 100 μL of serum-free culture medium to each well and place it in the incubator for 1 hour to hydrate the basement membrane. Aspirate the liquid in the upper chamber and adjust the cell density of the IOMM-Lee single cell suspension in the logarithmic growth phase to 2×10 cells using serum-free DMEM medium. 5 Cells were cultured at 400 μg / mL. 200 μL of the cell suspension was added to a Transwell chamber. The lower culture medium was DMEM supplemented with 20% FBS. Compound (I) was added to the chamber at various concentrations: 0 μM, 0.5 μM, 1 μM, and 2 μM. After 24 hours of incubation, the cells were removed with a cotton swab and fixed with 4% paraformaldehyde for 20 minutes. The cells were then washed three times with PBS and stained with crystal violet for 30 minutes. The cells were then observed and counted under a microscope.

[0085] (2) Referring to the experimental method (1), the concentrations of the compound (I) treatment group were changed to 2 μM, 5 μM and 10 μM, respectively, and the human meningioma cell lines used were IOMM-Lee or CH157-MN.

[0086] 2. Experimental Results

[0087] The experimental results showed that after 24 hours of treatment of IOMM-Lee cells or CH157-MN cells with different concentrations of compound (I), the invasion rate decreased significantly with increasing concentration of compound (I), as shown in Figure 4 (Figure 4-1, Figure 4-2, and Figure 4-3). These results indicate that compound (I) has a significant inhibitory effect on the invasion of meningioma cells IOMM-Lee and CH157-MN in a dose-dependent manner.

[0088] Example 5: Effect of Compound (I) on Apoptosis of Human Meningioma Cell Lines

[0089] 1. Experimental Methods

[0090] (1) Inoculate a single IOMM-Lee cell suspension in the logarithmic growth phase into a 6-well plate at a cell density of 5×10 5 / ml, cultured overnight, divided into blank control group and different concentrations of compound (I) treatment groups, the concentrations of compound (I) were 1μM, 5μM, 10μM, respectively, 3-4 replicates were set, and different concentrations of compound (I) were added for 24h. The blank control group and the compound (I) treatment group were collected (1-10)×10 5 Wash cells with PBS and centrifuge. Resuspend cells in 500 μl of 1× Binding Buffer. Add 5 μl of Annexin V-FITC and 10 μl of PI to each tube. Gently vortex to mix, incubate at room temperature in the dark for 5 minutes, and then analyze by flow cytometry.

[0091] (2) Referring to the experimental method (1), the concentrations of the compound (I) treatment group were changed to 2 μM, 5 μM and 10 μM, respectively, and the human meningioma cell lines used were IOMM-Lee or CH157-MN.

[0092] 2. Experimental Results

[0093] The experimental results showed that compared with the blank control group, after 24 hours of treatment of IOMM-Lee cells or CH157-MN cells with different concentrations of compound (I), the total apoptotic rate increased with increasing concentration of compound (I), with a significant increase in late apoptosis, as shown in Figure 5 (Figures 5-1, 5-2, and 5-3). These results indicate that compound (I) has a significant promoting effect on apoptosis in meningioma cells IOMM-Lee and CH157-MN.

[0094] Example 6: Typical clinical cases

[0095] Compound (I) is currently undergoing Phase I clinical trials and has conducted preliminary efficacy evaluation on patients with meningioma.

[0096] Drug: Compound (I) oral solution, 10-25 mg / day, once a day.

[0097] Dosing regimen for subjects: Single-dose phase: A single dose is initiated upon enrollment, followed by 3 days of observation; Multiple-dose phase: 21 consecutive days of administration. If no serious adverse events occur, subsequent treatment cycles can be initiated, each lasting 21 days, until disease progression (PD) or unacceptable toxicity occurs.

[0098] The tumor assessment adopts the "RANO meningioma tumor assessment criteria" (Huang, Raymond Y et al. Proposed response assessment and endpoints for meningioma clinical trials: report from the Response Assessment in Neuro-Oncology Working Group. Neuro-oncology vol. 21, 1 (2019): 26-36. doi: 10.1093 / neuonc / noy137) to evaluate the efficacy. The enhanced tumor lesions are selected as target lesions according to the standards, and they are accurately measured. The changes in the measured values ​​are compared to confirm the therapeutic efficacy of the target lesion tumor. All tumor lesions that are not selected as target lesions are recorded as non-target lesions. Non-target lesions are observed at each tumor assessment, and their changes are described and compared. In addition, the improvement and deterioration of the patient's clinical symptoms and the changes in the dosage of steroid hormones for symptomatic treatment of neurological symptoms will also be used as evaluation indicators in tumor assessment. Tumor assessments were performed at baseline before the first dose and every 6 weeks during treatment until disease progression (PD) or initiation of new anti-tumor therapy.

[0099] A total of 34 patients with residual, recurrent or progressive meningioma after surgery received compound (I) treatment. Most patients experienced tumor regression and stable or improved clinical symptoms after medication. As of January 16, 2023, tumor assessments were completed for 5 patients, including 2 PRs, 1 MR, and 2 SDs. As of February 2, 2024, 32 patients completed tumor assessments, including 4 PRs, 6 MRs, and 18 SDs. The objective response rate based on the RANO-meningioma standard assessment was 29.41%, the disease control rate was 82.4%, and 26 patients experienced tumor shrinkage after medication, accounting for 76.5% of all patients. Adverse events associated with compound (I) after administration were mostly mild and moderate, and those with an incidence greater than 20% included: hypertension, decreased platelet count, increased blood lactate dehydrogenase, sinus bradycardia, proteinuria, increased aspartate aminotransferase, increased alanine aminotransferase, hypokalemia, hyperlipidemia, decreased neutrophil count, decreased white blood cell count, diarrhea, and anemia.

[0100] Typical case 1:

[0101] A patient, female, 59 years old, disease diagnosis: anaplastic meningioma, pathological type: anaplastic (malignant) meningioma (CNS WHO grade III) infiltrating brain tissue.

[0102] On July 12, 2015, he underwent resection of the right middle intracranial meningioma. No subsequent chemoradiotherapy or targeted therapy was performed, and he was reviewed regularly. From November to December 2020, enhanced MRI of the head showed tumor progression. On January 18, 2021, he underwent meningioma resection. During the operation, the base of the tumor was found to be attached to the middle skull base. The tumor was 4*4*4.5cm. On July 4, 2022, he participated in a clinical trial of injectable honokiol liposomes (HK), and the last treatment time was July 31, 2022. On August 9, 2022, the patient's clinical symptoms worsened, and he was discharged from the group after comprehensive evaluation.

[0103] On August 16, 2022, the patient participated in the Phase I clinical trial of Compound (I). On August 31, 2022, the patient started taking Compound (I) oral solution, dose: 15 mg, once a day.

[0104] Tumor efficacy evaluation: A single dose was initiated upon enrollment. No adverse events occurred after three days of observation. Multiple doses were then initiated, and two cycles of treatment were completed. The subject experienced significant relief of headache and pain. A head MRI showed significant tumor reduction (56% reduction in the C2 target lesion), with no heal or progression of non-target lesions. The patient's clinical status was stable or improved without the use of hormones, resulting in an overall response rate (PR). No adverse events were reported during treatment.

[0105] Typical case 2:

[0106] A patient, female, 46 years old, disease diagnosis: meningioma (fibrous type), pathological type: meningioma (fibrous type), WHO grade I.

[0107] The patient underwent surgery on May 4, 2017. Another surgery was performed in June 2020, and Gamma Knife radiosurgery was performed in August 2020 and March 2021. On March 22, 2022, the patient received intravenous camrelizumab, but was intolerant and had vision loss, leading to gradual blindness.

[0108] On November 7, 2022, the patient participated in the Phase I clinical trial of Compound (I). On November 16, 2022, the patient started taking Compound (I) oral solution at a dose of 15 mg once daily. Baseline physical examination showed blindness in both eyes, with only light perception.

[0109] Tumor efficacy evaluation: A single dose was initiated after enrollment. No adverse events occurred after three days of observation. Multiple doses were then initiated. After two cycles, the subject returned to the hospital for a visit and reported vision recovery and regained self-care ability. Brain MRI showed significant tumor reduction (36% reduction in the C2 target lesion), no non-target lesions, no steroid use, and stable or improved clinical status. The overall efficacy was assessed as MRI. The main adverse event during treatment was mild to moderate hypertension, which resolved after medication was suspended.

[0110] Typical case 3:

[0111] A patient, female, 50 years old, disease diagnosis: meningioma.

[0112] In May 2018, he underwent right parietal-occipital tumor resection. Postoperative pathology revealed a grade III anaplastic meningioma, and he underwent concurrent chemoradiotherapy. In March 2021, he underwent a second surgery due to tumor recurrence. Postoperative pathology revealed a grade II atypical meningioma, and no further chemoradiotherapy was performed. In September 2022, he underwent a third right parietal-occipital craniotomy. Postoperative pathology confirmed a diagnosis of malignant meningioma.

[0113] On October 17, 2022, the patient participated in the Phase I clinical trial of Compound (I). On October 26, 2022, the patient started taking Compound (I) oral solution, dose: 15 mg, orally once daily.

[0114] Tumor efficacy evaluation: Head MRI showed significant tumor reduction (56% reduction in the C2 target lesion), no non-target lesions, no steroid use, and stable or improved clinical status, resulting in an overall response rate (PR). The main adverse event during treatment was mild to moderate hypertension, which resolved with the use of antihypertensive medication.

[0115] Typical case 4:

[0116] A patient, female, 55 years old, disease diagnosis: meningioma.

[0117] In May 2021, a meningioma was diagnosed and a right frontal open meningioma resection was performed. In June 2022, a second surgery was performed due to recurrence. Postoperative pathology showed an atypical meningioma, WHO grade II. Gamma knife radiotherapy was performed after surgery and no further treatment was received.

[0118] On May 5, 2023, he participated in the Phase I clinical trial of Compound (I). On May 11, 2023, he started taking the oral solution of Compound (I), dose: 20 mg, orally once a day.

[0119] Tumor efficacy evaluation: During the screening period, the patient demonstrated an 18x12mm irregular ring-shaped enhancement in the right superior sagittal sinus of the parietal lobe. After six cycles of treatment, a head MRI showed tumor shrinkage (61.47% reduction in the target lesion), with the non-target lesion in the right parietal lobe remaining stable. Without the use of steroids, the patient's clinical condition was stable or improved, resulting in an overall response rate (PR). The target lesion continued to shrink in subsequent efficacy evaluations. The patient has been taking the drug for nine months and is continuing treatment. Major adverse events during treatment were mild proteinuria, decreased platelet count, decreased neutrophil count, decreased white blood cell count, and elevated alanine aminotransferase, which resolved with symptomatic treatment.

[0120] Typical case 5:

[0121] A patient, male, 45 years old, disease diagnosis: meningioma.

[0122] In 2017, an abnormality in the left frontotemporal region was discovered. On September 14, 2017, the patient underwent the first surgery to remove the left frontotemporal mass. Postoperative pathology revealed a meningothelial meningioma, WHO grade I. In July 2019, the patient underwent a recurrence of the left temporal meningioma. Postoperative pathology revealed an atypical meningioma, WHO grade II. In April 2020, the patient underwent a third combined craniocervical-facial radical resection. Postoperative pathology revealed an atypical meningioma, WHO grade II. Postoperative chemoradiotherapy was then initiated. On December 29, 2022, the patient underwent a fourth brain lesion resection. Postoperative pathology revealed a papillary meningioma, WHO grade III. In August 2023, the patient underwent stereotactic radiotherapy (20 GUy / 4fx).

[0123] On October 19, 2023, he participated in the Phase I clinical trial of Compound (I). On October 25, 2023, he started taking the oral solution of Compound (I), dose: 25 mg, orally once a day.

[0124] Tumor efficacy evaluation: At screening, the patient had bilateral parafalcine enhancing lesions measuring 60 x 32 x 27 mm. After six cycles of treatment, a head MRI showed tumor reduction (target lesion reduction of 36.73%). Non-target lesions in the left temporal lobe and left orbit were stable. The patient was no longer receiving steroids, and her clinical status was stable or improved. The overall efficacy was assessed as MRI. Target lesions continued to shrink during subsequent efficacy evaluations. Major adverse events during treatment were mild hypertension, decreased platelet count, decreased neutrophil count, and elevated alanine aminotransferase, which resolved / recovered with symptomatic treatment.

[0125] The above preliminary clinical treatment data show that compound (I) can effectively treat meningioma and has good safety and tolerability.

[0126] The full names and Chinese names of the English abbreviations used in this application are as follows:

[0127] It is understood that although the inventions described herein are described in the aforementioned specific forms, these inventions are not limited to the specific contents described in these specific forms. It is obvious to those skilled in the art that various equivalent variations may be made to the technical features of the inventions described herein without departing from the spirit of the inventions described herein, and such variations should fall within the scope of the inventions.

Claims

1. Use of compound (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating meningioma, wherein the compound (I) has the following structure:

2. The use according to claim 1, wherein the meningioma is classified according to clinical pathological characteristics and includes the following meningiomas: WHO grade Ⅰ meningioma, WHO grade Ⅱ meningioma and WHO grade Ⅲ meningioma.

3. The use according to claim 2, wherein the WHO grade I meningioma is a benign meningioma, preferably one or more selected from meningothelial meningioma, fibrous meningioma, transitional meningioma, psammoma meningioma, angiomatous meningioma, microcystic meningioma, secretory meningioma, lymphoplasmacytic-enriched meningioma and metaplastic meningioma; and / or The WHO grade II meningioma is an atypical meningioma, preferably one or more selected from atypical meningioma, clear cell meningioma and chordal meningioma; and / or The WHO grade III meningioma is a malignant meningioma, preferably one or more selected from rhabdoid meningioma, papillary meningioma and anaplastic meningioma.

4. The use according to claim 1, wherein the meningioma is a meningioma that has failed or recurred after other treatments; the other treatments include surgery, radiotherapy and anti-tumor drug therapy.

5. The use according to claim 4, wherein the anti-tumor drugs include chemotherapeutic drugs, targeted therapy drugs and immunotherapy drugs.

6. The use according to claim 5, wherein the anti-tumor drug is selected from one or more of carrelizumab, bevacizumab, sunitinib, everolimus, etoposide, temozolomide, sirolimus, and magnolol.

7. A pharmaceutical composition for treating meningioma, comprising a therapeutically effective amount of compound (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier; Wherein the compound (I) has the following structure:

8. The pharmaceutical composition according to claim 7, wherein the pharmaceutical composition is an oral preparation; preferably an oral solution.

9. A method of treating a meningioma in an individual, comprising: administering a therapeutically effective amount of compound (I) or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7 or 8 to an individual suffering from meningioma; Wherein the compound (I) has the following structure:

10. The method of claim 9, wherein the therapeutically effective amount is: The daily dosage of compound (I) is 5 mg to 30 mg, preferably 10 mg to 25 mg, more preferably 10 mg to 20 mg or 15 mg to 25 mg, further preferably 10 mg, 15 mg, 20 mg or 25 mg.

11. The method according to claim 10, wherein compound (I) is administered as a single dose once a day, or is administered in divided doses or at intervals per day, preferably as a single dose once a day.

12. The method of claim 10, wherein the administration cycle of compound (I) is 21 consecutive days, and optionally, the administration can be repeated for multiple cycles.

13. The method of claim 9, wherein the meningioma is classified according to clinical pathological characteristics to include the following meningiomas: WHO grade Ⅰ meningioma, WHO grade Ⅱ meningioma and WHO grade Ⅲ meningioma, optionally, The WHO grade I meningioma is a benign meningioma, preferably one or more selected from meningothelial meningioma, fibrous meningioma, transitional meningioma, psammoma meningioma, angiomatous meningioma, microcystic meningioma, secretory meningioma, lymphoplasmacytic-enriched meningioma and metaplastic meningioma; and / or The WHO grade II meningioma is an atypical meningioma, preferably one or more selected from atypical meningioma, clear cell meningioma and chordal meningioma; and / or The WHO grade III meningioma is a malignant meningioma, preferably one or more selected from rhabdoid meningioma, papillary meningioma and anaplastic meningioma.

14. Compound (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound (I) or a pharmaceutically acceptable salt thereof, for use in treating meningioma, wherein the compound (I) has the following structure:

15. The compound (I) or a pharmaceutically acceptable salt thereof for treating meningioma according to claim 14, or a pharmaceutical composition comprising the compound (I) or a pharmaceutically acceptable salt thereof, wherein the meningioma is classified according to clinical pathological characteristics and includes the following meningiomas: WHO grade I meningioma, WHO grade II meningioma and WHO grade III meningioma, and optionally, The WHO grade I meningioma is a benign meningioma, preferably one or more selected from meningothelial meningioma, fibrous meningioma, transitional meningioma, psammoma meningioma, angiomatous meningioma, microcystic meningioma, secretory meningioma, lymphoplasmacytic-enriched meningioma and metaplastic meningioma; and / or The WHO grade II meningioma is an atypical meningioma, preferably one or more selected from atypical meningioma, clear cell meningioma and chordal meningioma; and / or The WHO grade III meningioma is a malignant meningioma, preferably one or more selected from rhabdoid meningioma, papillary meningioma and anaplastic meningioma.

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