Use of cariporide in the manufacture of a medicament for the treatment of mpnst

Abstract

The application belongs to the technical field of new application of drugs, and particularly relates to application of copanlisib in preparation of a drug for treating malignant peripheral nerve sheath tumor (MPNST). The application first applies copanlisib to the treatment of malignant peripheral nerve sheath tumor. Copanlisib, as a class I PI3K inhibitor, can directly target the key node, inhibit the malignant progression of MPNST from the molecular level, especially for NF1-related MPNST. At present, there is no literature report or clinical evidence suggesting that copanlisib can be used for the treatment of MPNST, and the application fills the technical gap of the drug in the field of MPNST treatment, and provides a new systemic treatment option for MPNST patients.

Description

Technical Field

[0001] This invention belongs to the field of new pharmaceutical application technology, specifically relating to the application of cupanicine in the preparation of a drug for the treatment of malignant peripheral nerve schwannoma (MPNST). Background Technology

[0002] Malignant peripheral nerve sheath tumor (MPNST) is a highly aggressive soft tissue sarcoma originating from peripheral nerve sheath cells or pre-existing neurofibromas. This tumor has a high rate of local recurrence and distant metastasis, particularly to the lungs, resulting in a poor overall prognosis with a 5-year survival rate of less than 50%. Currently, clinical treatment for MPNST remains focused on radical surgical resection; however, due to the tumor's frequent location in deep tissues or adjacent to important neurovascular structures, the complete resection rate is low. Radiotherapy and conventional chemotherapy offer limited benefits, and for patients with advanced, unresectable, or recurrent / metastatic tumors, there is a lack of effective systemic treatment strategies, highlighting a pressing unmet clinical need for effective treatment.

[0003] From a molecular pathology perspective, approximately half of all MPNSTs occur in a neurofibromatosis type 1 (NF1) background. The neurofibromin encoded by the NF1 gene, as a RAS-GTPase activator protein (RAS-GAP), is responsible for hydrolyzing activated RAS-GTP into inactivated RAS-GDP, and is a negative regulator of the RAS signaling pathway. Loss of NF1 function leads to persistent RAS activation, which in turn drives abnormal activation of downstream pro-proliferative and anti-apoptotic pathways such as RAS / MAPK (RAF-MEK-ERK) and PI3K-AKT-mTOR. Furthermore, inactivation of tumor suppressor genes such as CDKN2A and TP53, as well as loss-of-function mutations in the multicomb repressor complex 2 (PRC2) core component SUZ12 or EED, further promote the malignant progression of benign precursor neurofibromas to MPNSTs. The aforementioned molecular events collectively constitute the complex signaling network of MPNST, among which the PI3K-AKT-mTOR pathway, as a key hub connecting RAS signaling with cellular metabolism and survival, plays a crucial role in the occurrence and maintenance of MPNST. Therefore, inhibitors targeting the PI3K signaling pathway hold promise for providing a new therapeutic breakthrough for MPNST.

[0004] Copanlisib is an intravenously administered class I PI3K inhibitor with potent inhibitory activity against the p110α and p110δ isoforms, as well as p110β and p110γ. This drug reversibly binds to the ATP-binding pocket of PI3K, blocking PI3K-mediated phosphatidylinositol-3,4,5-triphosphate (PIP3) production, thereby inhibiting downstream AKT phosphorylation and the activation of its effector molecules, ultimately suppressing tumor cell proliferation and inducing apoptosis. Currently, copanlisib is approved for the treatment of relapsed follicular lymphoma, but there is no evidence to support or suggest its use in the treatment of MPNST. Summary of the Invention

[0005] The purpose of this invention is to propose the use of cupanicine in the preparation of a medicament for the treatment of malignant peripheral nerve schwannoma (MPNST).

[0006] Technical solution:

[0007] The first aspect of this invention proposes the use of cupanixine in the preparation of a drug for treating MPNST.

[0008] Furthermore, the MPNST is a malignant peripheral nerve sheath tumor associated with neurofibromatosis type 1.

[0009] Furthermore, the cupanicine achieves therapeutic effects on MPNST by inhibiting the proliferation of malignant peripheral nerve sheath tumor cells.

[0010] Furthermore, the cupanixexi achieves therapeutic effects on MPNST by inhibiting the invasion and / or metastasis of malignant peripheral nerve sheath tumor cells.

[0011] A second aspect of the present invention provides a pharmaceutical composition for treating malignant peripheral nerve sheath tumors, comprising a therapeutically effective amount of cupanidine and a pharmaceutically acceptable carrier.

[0012] Furthermore, the pharmaceutical composition uses cupanixine as the sole active ingredient.

[0013] Furthermore, the pharmaceutical composition also contains at least one additional active ingredient selected from MEK inhibitors, mTOR inhibitors, PRMT5 inhibitors, or LSD1 inhibitors.

[0014] Beneficial effects:

[0015] This application marks the first time that copanlisib has been used to treat malignant peripheral nerve schwannomas (MPNST). As a class I PI3K inhibitor, copanlisib directly targets key nodes, inhibiting the malignant progression of MPNST at the molecular level, particularly NF1-related MPNST. Currently, there is no literature or clinical evidence suggesting the use of copanlisib for MPNST treatment. This application fills a technological gap in the field of MPNST treatment with this drug, providing MPNST patients with a novel systemic treatment option.

[0016] This application systematically evaluated the efficacy of cupanidine against MPNST using three culture models: a 2D adherent system, a Matrigel 3D system, and a GF996 suspended organoid system. The results showed that cupanidine exhibited a clear concentration-dependent inhibitory effect in all three systems, with an IC50 as low as 0.01 μM in the GF996 suspended organoid system, demonstrating extremely strong antitumor activity. Cross-validation across multiple systems ensured the comprehensiveness and reliability of the efficacy evaluation. Furthermore, cupanidine maintained a consistent concentration-dependent inhibitory effect in all three culture systems, and its anti-MPNST activity was unaffected by differences in culture systems, indicating that the drug's antitumor effect is robust. Combined with its known clinical pharmacokinetic characteristics and safety data, this application lays a solid experimental foundation for the clinical translation of cupanidine in the treatment of MPNST.

[0017] This application provides a potential effective treatment for highly aggressive, clinically unmet rare tumors, which has significant clinical and social value. Attached Figure Description

[0018] Figure 1 Morphological characterization of PNST and benign NF clinical samples and patient-derived organoids; in the figure, a is the clinical manifestation of MPNST, b is the resected specimen, c is the HE-stained image of clinical tissue, d is the HE immunohistochemical image of organoids, e is the S100 immunohistochemical image; f is the corresponding clinical manifestation of benign NF, g is gross, h is the HE-stained image of clinical tissue, i is the HE immunohistochemical image of organoids, and j is the S100 immunohistochemical image.

[0019] Figure 2 To show the exome consistency and differences between PNST clinical tissues and matched organoids, figure a shows filtered SNV / Indel overlap, b shows VAF comparison of 208 co-word variants, c shows the six-base substitution profile, and d shows NF1p.R997K and VAF and supporting reads, e is the CNV / LOH trajectory of paired samples normalized by the mode total copy number, and f is the multidimensional WES consensus summary; this analysis is tumor-only WES, without paired peripheral blood, therefore the relevant variants are not directly expressed as germline filtered somatic mutations.

[0020] Figure 3 Transcriptional program preservation and remodeling for PNST clinical tissues and matched organoids; Figure a shows the protein-coding gene expression relationships and five descriptive classifications; b shows the number and proportion of genes in each classification; c shows the expression of marker genes for Schwann lineage, immunity, endothelium, ECM / mesenchyma, and culture / inflammation programs; d shows the gene expression changes and medians of functional modules.

[0021] Figure 4 The image shows the drug sensitivity results for the GF996 suspension system (organoid) cultured in this way.

[0022] Figure 5 The image shows the drug susceptibility results for the Matrigel 3D culture system.

[0023] Figure 6 The image shows the drug susceptibility results for a 2D adherent culture system.

[0024] Figure 7 The results show the drug sensitivity of organoids; in the figure, a is a representative image of the 2D adherent system, Matrigel 3D system and GF996 suspension system (organoids) before and after treatment with Copanlisib 0.0l μM for 5 days; b is a heatmap of IC50 of 9 candidate drugs in the three culture systems, sorted by the geometric mean of available IC50.

[0025] Figure 8 The figure shows the preliminary results of invasion inhibition; in the figure, a is a schematic diagram of chip invasion from the cell sample addition channel on the right to the drug / culture medium channel on the left, and b is a representative time series image of the control group and the Copanlisib group from 0-60h. Detailed Implementation

[0026] To further illustrate the technical means and effects of this invention, the following description, in conjunction with embodiments and accompanying drawings, provides a further explanation of the invention. It is understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it.

[0027] Statistical analysis: Independent culture batches of each patient model were used as biological replicates, and the mean of technical replicates within each batch was included in the analysis. Invasive time-series data were analyzed using linear mixed models (continuous variables) or generalized estimating equations (non-normally distributed), with treatment, time, and interaction as fixed effects, and patient models and experimental batches as random intercepts. Beta coefficients or OR / RR and 95% CI were reported, avoiding reporting only P-values. Multiple comparisons were corrected using Tukey's method.

[0028] Quality control: At the sample level, the sampling site, necrosis rate, and transport time are recorded; at the model level, the number of passages, pellet formation rate, diameter, resuscitation viability, and pathological / genetic matching are recorded; at the data level, the original images, analysis parameters, outlier identification, and reasons for exclusion are preserved. Pathological scoring and quantitative image analysis are blinded, with information hidden by a third-party encoding team.

[0029] Example 1: Clinical Tissue Acquisition and Pathological Confirmation

[0030] This study employed an exploratory design of "existing single-case pilot-continuous expansion-time-series revalidation". One previously identified MPNST clinical tissue-organoid paired model served as a methodological pilot case and was not used alone to determine biomarker thresholds. Approximately 10-15 pathologically confirmed NF1-related MPNST fresh tissue specimens were consecutively included (aiming to successfully establish at least 6 organoids capable of completing the full evaluation process), and approximately 8-12 localized benign NF specimens (aiming to successfully establish at least 6). Each patient-derived model was considered an independent biological sample, and each model underwent at least 3 experimental batches independently, with at least 3 technical replicates for each concentration.

[0031] Inclusion criteria included: pathologically confirmed MPNST or localized benign NF; availability of sufficient fresh tissue; complete clinicopathological data; signed informed consent and compliance with ethical requirements; peripheral blood collection was conducted concurrently for newly added cases whenever possible to differentiate between germline and tumor-related variants; in the absence of paired blood, the term "tumor single-sample filtered variant" was explicitly used, and information such as age, sex, NF1 clinical history, tumor location, grade, previous treatment, and sample necrosis rate was recorded. Patients with severe necrosis, contamination, insufficient tissue volume, or inability to form stable organoids were excluded from functional association analysis, but their modeling success rate and failure reasons were included in the statistics.

[0032] Example 2 Construction of organoids from malignant neurofibroma

[0033] Fresh tissues were transported to the laboratory within a specified timeframe, where they were aseptically mechanically minced and enzymatically digested to obtain cell / tissue aggregates. These were cultured in the GF996 suspension system (organoids) and the Matrigel 3D system, with short-term 2D culture retained as a methodological control if necessary. The number of inoculated cells, time to aggregate formation, aggregate formation rate, diameter, roundness, growth curve, passage stability, cryopreservation recovery rate, and contamination status were recorded. Antimicrobial susceptibility testing prioritized early-generation organoids, and was limited to culture times and diameter ranges with similar diameters.

[0034] The method for constructing organoids from malignant neurofibromas is as follows:

[0035] Step 1: Obtain fresh tumor tissue from a patient with malignant peripheral nerve sheath tumor, and perform sterile cleaning, removal of necrotic tissue and mechanical shearing on the fresh tumor tissue;

[0036] Tissue processing: The freshly removed tissue was cleaned twice with tissue cleaning solution to remove surface blood; it was then transferred to a 60mm culture dish, and the tissue was separated into 3-5mm pieces using sterile ophthalmic scissors and forceps. 3 Collect small tissue samples into 1.5mL EP tubes, 2-3 pieces per tube. Use ophthalmic scissors to cut the tissue into small pieces of approximately 0.5-1mm. 3 .

[0037] Step 2: Perform enzymatic digestion on the shredded tumor tissue to obtain a cell suspension containing malignant peripheral nerve sheath tumor cells;

[0038] Add enzyme digest I (catalog number D001-1, Suzhou Jiyan Biomedical Technology Co., Ltd.) to an EP tube to resuspend the tissue, transfer it to a new 60mm culture dish, and incubate according to 1cm... 3 Add 5-6 mL of enzyme digestion solution I to the tissue of the desired size. Incubate at 37°C for enzymatic digestion; every 10 minutes, use a Pasteur pipette to gently pipette the tissue to promote digestion; observe the dissociation of the tissue block and the number of detached cells under a microscope during this period.

[0039] Step 3: Filter, centrifuge, and resuspend the cell suspension to obtain cells to be inoculated;

[0040] After most of the tissue has been digested and a large number of cells have detached, tissue washing solution (B002, Suzhou Jiyan Biomedical Technology Co., Ltd.) is added to stop the digestion. All the liquid after digestion is transferred to a 15mL centrifuge tube and centrifuged horizontally at 1500-2000rpm to collect all cells. The cells are washed twice with tissue washing solution to remove residual digestion solution I.

[0041] Resuspend the cell pellet in tissue washing solution, let stand for 10 seconds to allow any undissociated fibrous tissue to settle, then transfer the supernatant cell suspension to a 100 μm cell sieve placed in a 50 mL centrifuge tube beforehand. Transfer the filtered cells to a 15 mL centrifuge tube and collect the cells by horizontal centrifugation at 1500-2000 rpm.

[0042] The collected cells were seeded into 6-well cell culture plates. 2.5 mL of NFT (Suzhou Jiyan Biomedical Technology Co., Ltd.) medium specifically for malignant neurofibroma was added. The plates were incubated statically for 2-3 days.

[0043] Once the cells have fully adhered to the culture medium, replace it with fresh medium, 2.5 mL each time. Change the medium every 2-3 days.

[0044] Once the cells reach 80% fusion, they can be passaged or directly used to construct organoids.

[0045] Wash adherent cells once with sterile PBS, add 1.5 mL of enzyme digestion solution II (D002, Suzhou Jiyan Biomedical Technology Co., Ltd.), and incubate at 37°C for 3-5 minutes. After the cells are completely dissociated, add 2 mL of culture medium, gently pipette to promote cell dissociation, collect the cells in a 15 mL centrifuge tube, and centrifuge horizontally at 1500-2000 rpm to collect the cell pellet.

[0046] Remove the supernatant and resuspend the cell pellet in 1 mL of culture medium.

[0047] Take 10 μL of cell suspension and mix it with an equal volume of AOPI. Count the number of viable cells using a cell counter.

[0048] Step 4: The cells to be inoculated are seeded into a micron-scale low-adsorption U-bottom well array culture system for three-dimensional culture, so that the cells self-aggregate to form organoids derived from patients with malignant peripheral nerve sheath tumors.

[0049] Each organoid contains 500-2000 cells seeded into a 96-well high-throughput 3D cell culture assay plate (GF996, Suzhou Jiyan Biomedical Technology Co., Ltd.). After promoting cell aggregation, the plates are incubated at 37°C for 24-48 hours to obtain organoids. Simultaneously, the cells are cultured using the Matrigel 3D system, with short-term 2D culture retained as a methodological control if necessary.

[0050] Primary tissue and organoids were subjected to HE and S100 assays. Vimettin and other indicators were supplemented based on pathological characteristics. Results are shown in [Table missing]. Figure 1All pathological images were recorded at consistent magnification and scale. Cell morphology, arrangement, necrosis, proliferation, and nerve sheath lineage phenotype were independently evaluated by two observers unaware of the drug sensitivity results. Benign NF was used as a reference for non-malignant nerve sheath origin and was not interpreted as a precursor to malignancy or a paired lesion with MPNST.

[0051] Depend on Figure 1 As can be seen, this embodiment has established organoids from fresh tumor tissue of MPNST patients and obtained corresponding clinical manifestations, resected specimens, primary tissue HE, organoid HE, and S100 immunohistochemical images. This embodiment also established organoids for localized benign NF, completing clinical, gross, primary tissue HE, and organoid HE / S100 recordings. These results demonstrate that this embodiment has established a complete chain from patient tissue collection and organoid construction to basic pathological evaluation, and also provides a sample basis for subsequent parallel functional evaluation of MPNST and non-malignant NF.

[0052] Example 3: Matching analysis of paired WES and RNA-seq

[0053] 3.1 Paired WES Analysis

[0054] WES will use primary tissue-based organ pairing as the analysis unit, comparing SNV / Indel overlap rates, shared variant VAF six-base substitution patterns, CNV / LOH chromosomal localization, STR homology, and key gene status such as NF1, TP53, CDKN2A, and SUZ12. Newly paired peripheral blood cases will be filtered for germline mutations. For single-sample tumor analysis, population frequency functional annotation and purity / ploidy background will be carefully interpreted, and mutations will not be directly defined as somatic mutations. Models with good shared subsets but significant changes in individual key variant VAFs will be recorded as clonal shifts and subjected to sensitivity analysis during drug sensitivity interpretation. Results are shown below. Figure 2 .

[0055] Figure 2 The results showed that the constructed MPNST organoids were highly consistent with the primary tumor tissue in terms of core driver gene mutations (NF1, TP53), mutation allele frequency distribution (Pearson correlation coefficient r = 0.974), and whole-genome mutation spectrum, confirming the fidelity of this patient-derived organoid model at the molecular level.

[0056] 3.2 RNA-seq matching analysis

[0057] RNA-seq employed a unified workflow for library preparation, sequencing depth, and data quality control. First, overall expression correlation, sample clustering, and batch effects were evaluated. Second, protein-coding genes were categorized according to pre-defined thresholds into low expression / unclassified, stable retention, retention but remodeling, tissue enrichment / organoid loss, and organoid enrichment / acquisition. Third, pre-defined modules such as Schwann lineage, immunity, endothelial, ECM / mesenchymal culture / inflammation, proliferation, and hypoxia / stress were described. Pre-treatment biomarker screening prioritized features detectable in primary tissues and stably retained in organoids. The culture acquisition procedure was primarily used as a model bias indicator or covariate. Results are shown below. Figure 3 .

[0058] Figure 3 The results showed that although in vitro culture led to the downregulation of some microenvironment-related genes (such as C1QA, HLA-DRA, and COL1A1), the organoids still closely clustered with their source tissues at the global transcriptome level, and the core neural sheath lineage marker SOX10 and the autonomous transcriptional program of tumor cells were preserved. These results confirm that the organoid model constructed in this application effectively removes the confounding interference of the in vivo microenvironment on drug efficacy evaluation, and can more realistically reflect the inherent drug sensitivity of MPNST tumor cells to Copanlisib, providing a purer and more personalized tumor biology model for subsequent multi-system drug efficacy comparisons.

[0059] Example 4: Evaluation of Copanlisib Multi-System Dose-Response and Therapeutic Window

[0060] Organoids with stable morphology and in the logarithmic growth phase were selected. A solvent control and a copanlisib concentration gradient were set up, covering five concentration points from 0.001 to 100 μM. Viability was assessed using Cell Titer-Glo 3D or a validated equivalent method after a uniform treatment time, with simultaneous bright-field imaging. A four-parameter logistic model was used to fit the dose-response curves, and the IC50 was calculated. Results are shown below. Figure 4-6 .

[0061] Figure 4-6 Drug sensitivity results were presented in three systems. Bright-field imaging showed that with increasing drug concentration, organoids / cells in all three systems exhibited varying degrees of morphological damage and number reduction, with the GF996 suspension system (organoids) showing the most significant spheroid fragmentation at a concentration of 100 μM. The dose-response curves all exhibited typical S-shaped fits, but significant differences in IC50 were observed among the different systems. Figure 4 Among them, the GF996 suspension system (organoid) showed the highest sensitivity, with an IC50 of approximately 0.01 μM, and the dose-response curve shifted to the left most significantly, indicating that it had the highest sensitivity. Figure 5, the Matrigel 3D system follows, with an IC50 of approximately 0.01 μM, but the curve shifts to the right to a greater extent than that of GF996; Figure 6 , the 2D adherent system has the lowest sensitivity, with an IC50 of approximately 0.13 μM. Combining the results of viability curves and morphological observations, Copanlisib exhibits the strongest antitumor activity in the GF996 suspension organoid system, and its mechanism is further verified by proliferation inhibition and apoptosis induction assays.

[0062] The above results indicate that the traditional 2D adherent system screening model may underestimate the actual efficacy of this drug, while the GF996 suspension organoid system can better reflect the true killing effect of the drug in the suspension state than the Matrigel 3D culture system and the 2D adherent system. More importantly, Copanlisib exhibits a half-maximal inhibitory concentration as low as 0.01 μM in the GF996 organoid model, and shows a clear concentration-dependent inhibitory effect in all three culture systems, confirming that its anti-MPNST activity is not affected by differences in culture systems, with good cross-system stability and universality. The above findings provide direct in vitro pharmacodynamic evidence for the application of Copanlisib in the preparation of a medicament for treating malignant peripheral nerve sheath tumor (MPNST), and also lay a foundation for subsequent in vivo pharmacodynamic verification and determination of the therapeutic window.

[0063] Example 5

[0064] Benign NF organoids use the same batch of drugs, concentrations and detection procedures. The in vitro selectivity is described by the IC50 values of MPNST and NF, and benign NF is not directly used as a substitute for normal tissue in safety evaluation. Paired or mixed effect analysis is adopted for different culture systems to distinguish the patient model effect, the culture system effect and the interaction between the two. The independent drug screening results of Pirarubicin, Onalespib, Linsitinib, Anlotinib and the like are only used to illustrate that the platform can distinguish different responses, and are not mixed with the main mechanism of Copanlisib and the chip invasion assay. The results are shown in Figure 7 a-b.

[0065] Figure 7 a shows representative morphological images before treatment and after 5 days of treatment with Copanlisib (0.01 μM) in the 2D adherent system, Matrigel 3D system and GF996 suspension system. In Figure 7 b, Pirarubicin, Onalespib, Linsitinib, Ifosfamide, Tazemetostat, Selumetinib and Larotrectinib in the heat map are only used as independent multi-drug screening references, showing that the platform can distinguish different responses; Figure 7 ​As shown in b, the IC50 values ​​of Copanlisib were approximately 0.13, 0.01, and 0.01 μM, respectively, and the IC50 of Anlotinib was 0.2423 μM, obtained from an independent GF996 experiment. The 2D adherent culture system and Matrigel 3D culture have not yet been tested. These results indicate that the MPNST organoids constructed using the GF996 system in this embodiment can grow stably without Matrigel embedding and exhibit quantifiable inhibitory responses to targeted drugs. Compared to 2D adherent culture, GF996 suspension culture preserves cell-cell contact and three-dimensional spatial structure, avoiding interference from changes in cell polarity and abnormal activation of adhesion signals on drug efficacy evaluation. Compared to the Matrigel 3D system, GF996 does not require exogenous Matrigel, is simpler to operate, has more standardized culture conditions, and exhibits smaller batch-to-batch variations, making it more suitable for high-throughput drug screening and cross-system efficacy comparisons. Based on the aforementioned organoid model, Copanlisib exhibited significant in vitro inhibitory activity in MPNST (IC50 as low as 0.01 μM), and its efficacy remained consistent across different culture systems, indicating a clear application potential for this drug in MPNST. These results validate the feasibility and advantages of the GF996 organoid model constructed in this application for pharmacodynamic evaluation and provide direct experimental evidence for the application of Copanlisib in the preparation of drugs for the treatment of malignant peripheral nerve schwannomas (MPNST).

[0066] Example 6: Quantitative Analysis of Organ-on-a-Chip Invasions

[0067] According to the final Figure 8 As shown in Figure a, organoid-derived cells were placed in the right-hand sample loading channel of the chip, with the middle channel serving as a collagen matrix barrier, and the left-hand channel for adding drugs / culture medium. A solvent control, low-to-medium doses of Copanlisib, and an orthogonal PI3K inhibitor group with a clearly defined mechanism were set up. Dose that maintained an overall viability of at least 80% of the control after treatment was prioritized to evaluate the relatively independent anti-invasive effect. A dose close to the IC50 was also set up to observe the comprehensive tumor-suppressive effect. Continuous imaging was performed at 0, 12, 24, 36, 48, and 60 hours, and extended to 72 hours if necessary.

[0068] Cell body area, invasion area, maximum invasion distance, average expansion rate, and number of branches were calculated using a fixed threshold or semi-automatic segmentation process. Image analysts blinded the processed groups. Each model underwent at least three independent microarray experiments, each including multiple evaluable fields of view. The invasion inhibition rate was interpreted in conjunction with concurrent cell viability, proliferation, and apoptosis. If a decrease in invasion only occurred under obvious cell death conditions, it was not considered a relatively independent anti-invasion effect.

[0069] Depend on Figure 8As shown in b, the control group exhibited significant cross-barrier migration from 36 hours onwards, and the migration distance and number of migrating cells increased in a time-dependent manner with prolonged culture time (48 hours, 60 hours), suggesting that MPNST cells possess a strong spontaneous invasive ability. The copanlisib-treated group showed no significant cross-matrix migration at any time point (36-60 hours), and even at 60 hours, only a very small number of cells crossed the collagen barrier, indicating that copanlisib significantly inhibited the invasive ability of MPNST cells. These results further suggest that copanlisib may affect the migration and invasion functions of tumor cells by inhibiting the PI3K-AKT signaling pathway.

[0070] The pathogenesis of MPNST involves the abnormal activation of multiple signaling pathways. Loss of NF1 function leads to persistent activation of the RAS signaling pathway, which in turn drives the abnormal activation of two key downstream pathways: RAF-MEK-ERK (MAPK) and PI3K-AKT-mTOR. Therefore, in some embodiments of this application, the pharmaceutical composition may further comprise at least one additional active ingredient selected from MEK inhibitors, mTOR inhibitors, PRMT5 inhibitors, or LSD1 inhibitors.

[0071] The above description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the concept described herein through the above teachings or related technologies or knowledge. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be within the protection scope of the appended claims.

Claims

1. Use of cupanlisib in the manufacture of a medicament for treating MPNST.

2. Use of bleperide according to claim 1 for the preparation of a medicament for the treatment of MPNST, characterized in that, The MPNST is a malignant peripheral nerve sheath tumor associated with neurofibromatosis type 1.

3. Use of bleperide for the manufacture of a medicament for the treatment of MPNST according to claim 1, characterized in that, The cupanlisib achieves the treatment of MPNST by inhibiting the proliferation of malignant peripheral nerve sheath tumor cells.

4. Use of bleperide according to claim 1 for the preparation of a medicament for the treatment of MPNST, characterized in that, The cupanlisib achieves the treatment of MPNST by inhibiting the invasion and / or metastasis of malignant peripheral nerve sheath tumor cells.

5. A pharmaceutical composition for treating malignant peripheral nerve sheath tumor, characterized by, A pharmaceutical composition comprising a therapeutically effective amount of cupanlisib and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition for use in the treatment of malignant peripheral nerve sheath tumors according to claim 5, characterized in that, The pharmaceutical composition has cupanlisib as the only active ingredient.

7. The pharmaceutical composition for use in the treatment of malignant peripheral nerve sheath tumors according to claim 5, characterized in that, The pharmaceutical composition further comprises at least one additional active ingredient selected from a MEK inhibitor, an mTOR inhibitor, a PRMT5 inhibitor, or an LSD1 inhibitor.

Images