A pharmaceutical combination for treating non-small cell lung cancer

Abstract

This disclosure provides a pharmaceutical combination for treating non-small cell lung cancer, comprising a therapeutically effective amount of a PIN1 inhibitor and a therapeutically effective amount of neratinib.

Description

Technical Field

[0001] This disclosure pertains to the field of pharmaceutical technology and relates to a drug combination for the treatment of non-small cell lung cancer. Background Technology

[0002] Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer cases and remains the leading cause of cancer-related deaths. Despite significant advances in targeted therapy, most patients eventually develop treatment resistance, severely limiting long-term survival. ERBB family receptor tyrosine kinases, including EGFR, HER2, HER3, and HER4, form the core signaling network driving NSCLC tumorigenesis and maintaining its survival. Notably, these receptors are frequently co-expressed in lung tumors and function by forming highly active homodimers and heterodimers. The clinical efficacy of highly selective EGFR or HER2 inhibitors is limited because NSCLC cells can dynamically remodel their receptor network, bypassing single-target blockade through compensatory heterodimerization.

[0003] To overcome this receptor-level resistance, researchers have developed irreversible pan-ERBB tyrosine kinase inhibitors, such as neratinib, which can form covalent bonds within the ATP binding pocket of these receptors, thereby achieving sustained inhibition and preventing receptor reactivation. However, recent clinical trials evaluating neratinib for advanced lung cancer have shown limited objective response rates when used as monotherapy. Furthermore, dual therapy with drugs such as temsirolimus or trastuzumab has also failed to significantly improve overall patient outcomes, highlighting the profound impact of intrinsic and adaptive bypass mechanisms. Therefore, there is an urgent clinical need to enhance the therapeutic potential of neratinib in NSCLC. Summary of the Invention

[0004] This disclosure provides a drug combination for treating NSCLC that can effectively address the above-mentioned problems.

[0005] This disclosure is implemented as follows: In a first aspect, this disclosure provides a pharmaceutical combination comprising a therapeutically effective amount of a PIN1 inhibitor and a therapeutically effective amount of neratinib.

[0006] Secondly, this disclosure provides the use of PIN1 inhibitors in combination with neratinib in the preparation of drugs for inducing apoptosis in NSCLC cells.

[0007] Thirdly, this disclosure provides the use of PIN1 inhibitors in the preparation of drugs for enhancing the sensitivity of NSCLC cells to neratinib.

[0008] Compared with the prior art, this disclosure has the following beneficial effects: (1) Significant synergistic anti-tumor effect The combination of PIN1 inhibitors and neratinib can produce a significant synergistic anti-tumor effect, effectively inducing apoptosis in NSCLC cells and overcoming the efficacy limitations of neratinib monotherapy.

[0009] (2) Enhance the therapeutic potential of neratinib in NSCLC PIN1 inhibitors can significantly enhance the sensitivity of NSCLC cells to neratinib, providing a novel strategy for improving the therapeutic potential of neratinib in NSCLC. Attached Figure Description

[0010] To more clearly illustrate the technical solutions of the embodiments of this disclosure, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this disclosure and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0011] Figure 1 This is the result of Kaplan-Meier survival curve analysis, which divides stage I NSCLC patients into a low expression group (black, n=417) and a high expression group (red, n=239) based on PIN1 mRNA expression levels.

[0012] Figure 2 This is a comparison of PIN1 protein levels in PIN1-WT and PIN1-KO NCI-H460 cells detected by Western blot, using β-actin as an internal control, to confirm the effectiveness of CRISPR / Cas9-mediated PIN1 gene knockout.

[0013] Figure 3 The cell viability scatter plot is obtained by high-throughput drug screening of PIN1-WT and PIN1-KO NCI-H460 cells using an FDA-approved targeted drug library (n=103 drugs). The red dashed line represents the 10% cell viability threshold.

[0014] Figure 4 This is a bar chart comparing the cell viability of PIN1-WT cells (blue bars) and PIN1-KO cells (red bars) after treatment with targeted drugs that reduced PIN1-KO cell viability to below the 10% threshold. Data are expressed as mean ± standard deviation, *p<0.05, ****p<0.0001.

[0015] Figure 5This is a ranking chart of the PIN1-WT / PIN1-KO cell viability ratio (WT-to-KO ratio) for each screened targeted drug.

[0016] Figure 6 The results are based on correlation analysis using the Cancer DepMap database. The left figure is a scatter plot of the area under the curve (AUC) sensitivity of PIN1 mRNA expression level (Log2(TPM+1)) and neratinib in 350 cancer cell lines (Pearson=0.301, p=8.89e-9). The right figure is a violin plot of neratinib AUC sensitivity for cell lines with low and high PIN1 expression (p<0.0001).

[0017] Figure 7 The results are the cell viability assays of NCI-H460 cells after treatment with different concentrations of neratinib (0, 2, 4, 6 μM) and the PIN1 inhibitor Sulfopin (0, 2, 4, 8 μM) as single or combined drugs. Data are expressed as mean ± standard deviation. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0018] Figure 8 The images show heatmaps of cell inhibition rates at different concentrations of neratinib and Sulfopin (top image) and two-dimensional contour plots of ZIP synergistic fractions calculated based on cell viability data (bottom image).

[0019] Figure 9 The left image shows a representative scatter plot of NCI-H460 cells treated with DMSO control, Sulfopin, neratinib, or a combination of both, and a quantitative statistical bar chart of the total percentage of apoptotic and necrotic cells in each treatment group, obtained by Annexin V / propidium iodide (PI) double staining flow cytometry. The right image shows a quantitative statistical bar chart of the percentage of apoptotic and necrotic cells in each treatment group. *p<0.05, **p<0.01.

[0020] Figure 10 These are representative images and quantitative analysis results of the long-term colony formation assay of NCI-H460 cells after treatment with Sulfopin, neratinib alone or in combination. **p<0.01, ***p<0.001, ns indicates no significant difference.

[0021] Figure 11The data include tumor growth curves of NCI-H460 xenografts in different treatment groups, tumor weight measurements of mice euthanized 12 days after treatment (Vehicle and Sulfopin groups n=5, neratinib and combination groups n=4), and weight monitoring results of mice in each group during treatment. Data are expressed as mean ± standard error. Statistical analysis was performed using an unpaired two-sided t-test. *p<0.05, ns indicates no significant difference.

[0022] Figure 12 In the diagram, A and C represent the KEGG pathway enrichment analysis results of phosphorylated proteins that were significantly downregulated relative to the DMSO control after NCI-H460 cells were treated with Sulfopin (A), neratinib (B), or a combination of both (C). D is a hierarchical clustering heatmap of the main downregulated phosphorylation sites in the MAPK pathway in the combined treatment group (Z-score represents relative phosphorylation intensity). EG is a Log2 fold waterfall plot of phosphorylation site changes in different treatment groups (E: Sulfopin vs. control; F: neratinib vs. control; G: combined treatment vs. control), marking the most significantly downregulated phosphorylation sites in the MAPK pathway in each group. H is a bar chart showing the enrichment of MAPK pathway proteins based on published PIN1 immunoprecipitation-mass spectrometry (IP-MS) data (Log2 fold change of GST-PIN1 relative to GST control).

[0023] Figure 13 The results are from a single-sample gene set enrichment analysis (ssGSEA) of primary tumor transcriptome data from the TCGA lung adenocarcinoma (LUAD) cohort. The samples were divided into a low PIN1 expression group (blue, n=300) and a high PIN1 expression group (red, n=301). A represents the gene set enrichment results for the KEGG pathway; B represents the gene set enrichment results for Gene Ontology Biology Processes (GOBP) and GNF2; and C represents the gene set enrichment results for the REACTOME pathway.

[0024] Figure 14 In this table, A represents the results of Western blot analysis of p-ERK1 / 2 and p-PAK2 protein levels in each treatment group. NCI-H460 cells were treated with DMSO, Daraxonrasib (1 μM), neratinib (4 μM), Sulfopin (8 μM), or a combination of neratinib and Sulfopin (two or three drugs), with β-actin as an internal control. B represents the results of Western blot analysis of p-PAK2 protein levels in NCI-H460 cells after treatment with increasing concentrations of Daraxonrasib (0, 50, 250, 500, 1000 nM).

[0025] Figure 15The images are representative scatter plots of NCI-H460 cells treated with DMSO, Daraxonrasib, Sulfopin + neratinib (dual or triple therapy), and flow cytometry using Annexin V / PI to detect apoptosis. They also include quantitative statistical bar charts of the percentage of apoptotic and necrotic cells. Data are expressed as mean ± standard deviation. *p<0.05, **p<0.01.

[0026] Figure 16 This is a schematic diagram of the downstream signaling bifurcation cascade pathway mechanism of the three-drug combination regimen: the ERBB signaling bifurcation is the classic RAS-ERK proliferation pathway (blue) and the anti-apoptotic RAC / Cdc42-PAK2-MAP3K3 survival pathway (pink). Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of the embodiments of this disclosure clearer, the technical solutions of the embodiments of this disclosure will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this disclosure, not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this disclosure without inventive effort are within the scope of protection of this disclosure. Therefore, the following detailed description of the embodiments of this disclosure provided in the accompanying drawings is not intended to limit the scope of the claimed disclosure, but merely represents selected embodiments of this disclosure. All other embodiments obtained by those skilled in the art based on the embodiments of this disclosure without inventive effort are within the scope of protection of this disclosure.

[0028] The “neratinib” mentioned in this disclosure refers to the compound Neratinib (CAS: 698387-09-6), whose chemical structure is shown in formula (I), including but not limited to its free base and its pharmaceutically acceptable salt.

[0029] (I) The “Daraxonrasib” mentioned in this disclosure refers to compound RMC-6236 (CAS: 2765081-21-6), whose chemical structure is shown in formula (II), including but not limited to its free base and its pharmaceutically acceptable salt.

[0030] (II) The “Sulfopin” mentioned in this disclosure refers to the compound PIN1-3 (CAS: 2451481-08-4), whose chemical structure is shown in formula (III), including but not limited to its free base and its pharmaceutically acceptable salt.

[0031] (III) The “pharmaceutically acceptable carriers or excipients” described in this disclosure include, but are not limited to, diluents, disintegrants, binders, lubricants, pH adjusters, isotonic adjusters, lyophilization protectants, solvents, surfactants, and preservatives, with specific examples being conventional choices in the art.

[0032] Example 1: Clinical relevance of PIN1 in NSCLC and the effect of PIN1 knockout on neratinib sensitivity Proline-directed phosphorylation of serine or threonine residues (pSer / Thr-Pro) is a central hub in cancer signal transduction, broadly regulating the stability and function of various oncoproteins and tumor suppressor proteins. This process is critically regulated by PIN1. PIN1 is a unique peptidyl-prolyl cis-trans isomerase that specifically recognizes phosphorylated Ser / Thr-Pro motifs and catalyzes conformational changes in its target proteins. Although PIN1 is frequently overexpressed in NSCLC and promotes tumorigenesis, its specific role in mediating resistance to pan-ERBB inhibitors, and whether inhibiting PIN1 can enhance the efficacy of the pan-ERBB inhibitor neratinib, remain unclear.

[0033] To explore the clinical significance and functional role of PIN1 in NSCLC, this publication analyzed transcriptomic datasets from independent NSCLC patient cohorts with clinical annotations. Kaplan-Meier survival analysis showed that in stage I NSCLC patients (low expression group n=417, high expression group n=239), high PIN1 mRNA expression was significantly associated with shortened overall survival (HR=1.58, 95% CI: 1.23–2.03, logrank P=0.00027). These results suggest that high PIN1 expression is a poor prognostic factor for NSCLC patients. Figure 1 ).

[0034] Furthermore, this disclosure describes the construction of a PIN1 gene knockout (PIN1-KO) NCI-H460 cell line using CRISPR / Cas9 gene editing technology. This cell line is an aggressive NSCLC cell line that is primarily resistant to neratinib and carries the KRAS Q61H mutation. Western blot analysis confirmed that PIN1 protein expression was completely eliminated in the knockout cells. Figure 2 Using PIN1 wild-type (PIN1-WT) NCI-H460 cells as a control, a high-throughput screening experiment was conducted using a drug library containing 103 FDA-approved targeted therapies. Figure 3As shown, in PIN1-WT cells, cell viability remained at a high level after most drug treatments; while in PIN1-KO cells, cell viability was significantly reduced after the same drug treatment, with a large number of data points below the 10% threshold, indicating that PIN1 knockout fundamentally increases the sensitivity of tumor cells to a variety of targeted therapies.

[0035] To further identify the drug with the most significant sensitizing effect, this disclosure includes a quantitative analysis of the screening results. For example... Figure 4 As shown, comparing the cell viability of PIN1-WT and PIN1-KO cells, among the tested drugs, the pan-ERBB inhibitor neratinib (marked in red) showed the most significant difference: neratinib treatment resulted in higher cell viability in PIN1-WT cells, while cell viability decreased significantly in PIN1-KO cells, with a statistically significant difference (****P<0.0001). Figure 5 As shown, neratinib ranked first among all tested drugs in terms of the PIN1-WT to PIN1-KO cell viability ratio, exhibiting the highest sensitization effect. This indicates that PIN1 activity is crucial for maintaining tumor cell survival signals under ERBB signaling blockade conditions.

[0036] The above results indicate that high PIN1 expression is not only associated with poor prognosis in NSCLC patients, but also that PIN1 deficiency can significantly enhance the sensitivity of NSCLC cells to the pan-ERBB inhibitor neratinib.

[0037] Example 2: Sulfopin combined with neratinib induces apoptosis in NSCLC cells To elucidate the relationship between PIN1 expression levels and neratinib sensitivity, this disclosure first analyzes the Cancer DepMap dataset. For example... Figure 6 As shown, among 350 cancer cell lines, the PIN1 mRNA expression level (Log2(TPM+1)) was significantly positively correlated with the area under the curve (AUC) of neratinib (Pearson=0.301, P=8.89e-9), indicating that higher PIN1 expression was associated with lower neratinib sensitivity; at the same time, the neratinib AUC sensitivity of the PIN1 low expression cell line group was significantly higher than that of the PIN1 high expression group (P<0.0001).

[0038] To validate the function and clarify whether PIN1 inhibition can enhance the antitumor efficacy of neratinib, this disclosure describes the treatment of NCI-H460 cells with the selective PIN1 inhibitor Sulfopin (0, 2, 4, 8 μM), neratinib (0, 2, 4, 6 μM), or a combination of both. Figure 7As shown, single-drug treatment produced only moderate cell viability inhibition, while the combination of Sulfopin and neratinib produced a significant, dose-dependent synergistic cell viability inhibition effect (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001). The ZIP synergistic score calculated based on cell viability data was 20.545 (…). Figure 8 This indicates that the two have a strong synergistic effect.

[0039] Further flow cytometry using Annexin V / PI double staining was used to detect cell apoptosis. The results showed that the apoptosis rates were low in the DMSO control group, the Sulfopin monotherapy group, and the neratinib monotherapy group, while the total percentage of apoptotic and necrotic cells in the Sulfopin + neratinib combination treatment group was significantly higher than that in any single-drug group (*P<0.05, **P<0.01). Figure 9 Furthermore, long-term colony formation experiments confirmed that the combined treatment had a significantly stronger inhibitory effect on colony formation potential than the single-drug treatment (**P<0.01, ***P<0.001). Figure 10 ).

[0040] To further evaluate the in vivo antitumor efficacy of the combined regimen of the two drugs, this disclosure established an NCI-H460 xenograft mouse model. NCI-H460 cells were subcutaneously inoculated into the lateral abdomen of 6-week-old female BALB / c nude mice. When the tumor diameter reached approximately 5 mm, the mice were randomly assigned to groups to begin treatment. Sulfopin was administered intraperitoneally (60 mg / kg, dissolved in 5% DMSO / D5W, daily); neratinib was administered orally by gavage (60 mg / kg, dissolved in 0.5% CMC-Na, daily). The dosage and regimen of each drug in the combination therapy group were the same as in the single-drug groups. Figure 11 As shown, compared with the control group (Vehicle group, n=5), the Sulfopin monotherapy group (n=5) and the neratinib monotherapy group (n=4) had limited inhibitory effects on tumor growth, while the Sulfopin and neratinib combination group (n=4) significantly inhibited tumor growth. Tumor weight was measured at euthanasia of mice 12 days after treatment, and the results showed that the tumor weight in the combination group was significantly lower than that in the monotherapy group (*P<0.05). Figure 11 (Middle). Weight monitoring results of mice in each group during treatment showed that the combined administration did not cause significant toxic side effects. Figure 11 Down).

[0041] The above results indicate that the combination of Sulfopin and neratinib produces a significant synergistic antitumor effect both in vitro and in vivo. This synergistic effect is mainly driven by a strong induction of apoptosis, thereby making NSCLC cells significantly sensitive to neratinib.

[0042] Example 3: Sulfopin and neratinib synergistically block the classical RAS-ERK and anti-apoptotic PAK2-MAP3K3 signaling pathways To elucidate the molecular mechanism of the synergistic effect of Sulfopin and neratinib, this disclosure presents a quantitative phosphorylated proteomics analysis of NCI-H460 cells. At a strict post-translational modification (PTM) localization probability threshold (0.75), approximately 13,000 phosphopeptides were reliably identified and quantified in each treatment group (CTR: DMSO control group; NER: neratinib monotherapy group; SUL: Sulfopin monotherapy group; SUL_NER: combination treatment group). KEGG pathway enrichment analysis showed that the MAPK signaling pathway was significantly enriched in the Sulfopin monotherapy, neratinib monotherapy, and both combination treatment groups, with the most significant downregulation in the combination treatment group. Figure 12 The AC of the combined treatment group was analyzed by hierarchical clustering of differentially phosphorylated sites in the MAPK pathway. The results showed that treatment with Sulfopin or neratinib alone only caused mild inhibition of phosphorylation levels in the MAPK pathway, while the combined treatment of the two drugs resulted in a significant synergistic reduction in multiple key phosphorylation nodes in the MAPK pathway. Figure 12 (D).

[0043] Further analysis of the Log2 fold changes of all phosphorylation sites revealed that the combination therapy synergistically reduced phosphorylation levels at corresponding sites on multiple key proteins in both the classical RAS-ERK pathway and the alternative PAK2-MAP3K3 pathway. These key proteins include, but are not limited to: NF1, BRAF, RPS6KA3 / 4, PAK1 / 2, and MAP2K2 / 3. Figure 12 To verify whether PIN1 has a direct regulatory effect on core proteins of the MAPK signaling pathway, this disclosure analyzes published PIN1 immunoprecipitation-mass spectrometry (IP-MS) datasets. The results show that there are direct physical interactions between PIN1 and multiple core proteins of the MAPK signaling network, with the top-ranking binding proteins including BRAF, RAF1, MAP3K2 / 3, MAP2K2, and PAK2 (E–G). Figure 12 H).

[0044] To further validate the clinical relevance of PIN1 to the MAPK pathway, this publication also analyzed transcriptomic data from the TCGA lung adenocarcinoma (LUAD) cohort, dividing the samples into a low PIN1 expression group (n=300) and a high PIN1 expression group (n=301). Single-sample gene set enrichment analysis (ssGSEA) results showed that at the KEGG pathway level, tumors with high PIN1 expression exhibited significant upregulation in the global MAPK signaling network (including the classical RAS-ERK branch and the anti-apoptotic PAK2-MAP3K3-ERK5 survival axis). Figure 13 At the GOBP and GNF2 gene set level, high expression of PIN1 was significantly associated with upregulation of the ERK1 / ERK2, ERK5 cascade pathways, and PAK2-specific functional network. Figure 13 In the REACTOME pathway, PIN1-overexpressing tumors showed significantly enhanced upstream ERK signaling and downstream ERK / MAPK target activation. Figure 13 (C).

[0045] The above results indicate that PIN1, as a core regulatory node, maintains the phosphorylation integrity of the classical RAS-ERK and alternative PAK2-MAP3K3 signaling pathways through physical interactions, while the combined treatment of Sulfopin and neratinib can effectively disrupt this regulatory network.

[0046] Example 4: Sulfopin combined with neratinib overcomes adaptive resistance by blocking PAK2 compensatory activation. To verify whether the combination of Sulfopin and neratinib blocked phosphorylation at key nodes in the RAS-ERK and PAK2-MAP3K3 signaling cascade, this disclosure used Western blot to detect the protein levels of p-PAK2 and p-ERK1 / 2. Specific treatment conditions were as follows: NCI-H460 cells were treated with DMSO, neratinib (4 μM), Sulfopin (8 μM), or a combination of neratinib and Sulfopin. Results showed that the combination of Sulfopin and neratinib significantly inhibited the levels of p-PAK2 and p-ERK1 / 2, indicating that this two-drug regimen can simultaneously target both the classical RAS-ERK and alternative PAK2-MAP3K3 signaling pathways. Figure 14 A).

[0047] Given that NCI-H460 cells carry the KRAS Q61H mutation, this disclosure further investigates the use of the pan-KRAS inhibitor Daraxonrasib as a baseline targeted therapy. Results showed that, unlike the combination of Sulfopin and neratinib, while Daraxonrasib monotherapy effectively eliminated p-ERK1 / 2, it caused a compensatory significant increase in p-PAK2 levels. Figure 14 (A). Further treatment of NCI-H460 cells with escalating concentrations of Daraxonrasib (0, 50, 250, 500, 1000 nM) showed a dose-dependent compensatory increase in p-PAK2 levels. Figure 14 The results (B) indicate that under the condition of KRAS blockade by Daraxonrasib, cells activated an adaptive survival feedback loop. Annexin V / PI double staining flow cytometry further showed that Daraxonrasib monotherapy failed to effectively induce apoptosis. Figure 15 ).

[0048] To clarify whether blocking adaptive PAK2 signaling can enhance the therapeutic effect of daraxonrasib and induce apoptosis, this disclosure further evaluated the effect of the combination of Sulfopin and neratinib on daraxonrasib resistance phenotype. The results showed that the combination of Sulfopin and neratinib significantly enhanced the therapeutic effect of daraxonrasib and induced a large number of apoptosis (*P<0.05, **P<0.01), effectively overcoming the intrinsic adaptive resistance exhibited by daraxonrasib monotherapy. Figure 15 ).

[0049] In summary, this disclosure proposes as follows: Figure 16 The illustrated mechanism model illustrates the synergistic mechanism by which the Sulfopin and neratinib combination regimen overcomes adaptive resistance to daraxonrasib: ERBB signaling branches into the classical RAS-ERK proliferation pathway (blue) and the anti-apoptotic RAC / Cdc42-PAK2-MAP3K3 survival pathway (pink). PIN1, as a key regulator, stabilizes multiple phosphorylation nodes in the two signal cascades through isomerization. The key limitation of daraxonrasib monotherapy in blocking KRAS is that while it effectively blocks RAS-ERK proliferation signaling, it cannot induce apoptosis and triggers compensatory activation of the PAK2-MAP3K3 anti-apoptotic pathway, thus leading to adaptive resistance. The Sulfopin and neratinib combination regimen, by simultaneously targeting the ERBB receptor and PIN1, can effectively block this compensatory PAK2-MAP3K3 anti-apoptotic cascade, inducing a large number of apoptosis, thereby overcoming the adaptive resistance induced by daraxonrasib monotherapy. The above results indicate that the combination of Sulfopin and neratinib provides a novel and effective treatment strategy for overcoming resistance in NSCLC, particularly NSCLC that has developed adaptive resistance to Daraxonrasib monotherapy.

[0050] Example 5: A pharmaceutical composition containing a PIN1 inhibitor and neratinib (two dosage forms combined) This embodiment provides a drug combination for treating NSCLC, consisting of the following two independent dosage forms: (1) Sulfopin lyophilized powder for injection (for intravenous injection) The formulation composition and dosage of each component of the lyophilized powder injection (for intravenous injection) are shown in Table 1.

[0051] Table 1. Formulation of Lyophilized Powder for Intravenous Injection

[0052] Preparation method: Sulfopin and mannitol were dissolved in water for injection and the volume was adjusted to 1 mL. After sterile filtration through a 0.22 μm filter membrane, the solution was dispensed into sterile vials and freeze-dried in a freeze dryer to prepare a lyophilized powder for injection. The solution was then sealed and stored. Before clinical use, the solution was reconstituted with water for injection and administered intravenously.

[0053] (2) Neratinib oral tablets The formulation composition and dosage of each component of neratinib oral tablets are shown in Table 2.

[0054] Table 2. Formulation of neratinib oral tablets

[0055] Preparation method: Lenatinib, microcrystalline cellulose, lactose, and croscarmellose sodium are mixed evenly in the above proportions. Magnesium stearate is added and the mixture is further mixed before being compressed into tablets. Each tablet contains 40 mg of the active ingredient lenatinib. For clinical use, it is administered orally.

[0056] Example 6: Two-drug kit for treating NSCLC This embodiment provides a drug kit for treating NSCLC, comprising: (a) First component: a formulation containing a therapeutically effective amount of the PIN1 inhibitor Sulfopin; Specifically, 50 mg of Sulfopin and 50 mg of mannitol were dissolved in water for injection, and the volume was adjusted to 1 mL. After sterile filtration through a 0.22 μm filter membrane, the solution was dispensed into a first independent container (sterile vial) and freeze-dried to prepare a lyophilized powder for injection.

[0057] (b) Second component: a formulation containing a therapeutically effective amount of neratinib; Specifically, 40 mg of natinib, 100 mg of microcrystalline cellulose, 60 mg of lactose, and 10 mg of croscarmellose sodium are mixed evenly, and 2 mg of magnesium stearate is added before being compressed into tablets and packaged in a second independent container (aluminum-plastic blister packaging).

[0058] The first component and the second component are physically independent formulations.

[0059] Instructions for use: The first component is reconstituted with water for injection and administered intravenously; the second component is administered orally. The two components may be administered to the subject simultaneously, separately, or sequentially, with the first component preferably administered before the second component, or both on the same day.

[0060] Recommended dosage (based on in vivo animal data of this disclosure): Sulfopin: 30–120 mg / kg body weight, preferably 60 mg / kg body weight; Lenatinib: 30–120 mg / kg body weight, preferably 60 mg / kg body weight.

[0061] The above description is merely a preferred embodiment of this disclosure and is not intended to limit the scope of this disclosure. Various modifications and variations can be made to this disclosure by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this disclosure should be included within the scope of protection of this disclosure.

Claims

1. A drug combination, characterized in that, The drug combination comprises a therapeutically effective amount of a PIN1 inhibitor and a therapeutically effective amount of neratinib.

2. The drug combination according to claim 1, characterized in that, The PIN1 inhibitor is Sulfopin, or a pharmaceutically acceptable salt, solvate, or prodrug of Sulfopin.

3. The drug combination according to claim 1 or 2, characterized in that, The drug combination is used to treat non-small cell lung cancer.

4. The drug combination according to claim 2, characterized in that, The Sulfopin is formulated as an injection, a lyophilized powder for injection, or an intravenous infusion; the neratinib is formulated as tablets, capsules, or granules.

5. The drug combination according to claim 2, characterized in that, The dosage of Sulfopin is 30–120 mg / kg body weight, and the dosage of neratinib is 30–120 mg / kg body weight; preferably, the dosage of both Sulfopin and neratinib is 60 mg / kg body weight.

6. Application of PIN1 inhibitors in combination with neratinib in the preparation of drugs for inducing apoptosis in non-small cell lung cancer cells.

7. The application according to claim 6, characterized in that, The non-small cell lung cancer cells are tumor cells that have developed primary or acquired resistance to neratinib monotherapy.

8. Application of PIN1 inhibitors in the preparation of drugs for enhancing the sensitivity of non-small cell lung cancer cells to neratinib.

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