A small molecule inhibitor targeting nid1 and applications thereof

Abstract

The application discloses a small-molecule inhibitor targeting NID1 and application, and relates to the technical field of biological medicines.The application selects W-5 hydrochloride as a small molecule for specifically inhibiting the function and activity of NID1 by molecular docking prediction and function verification from existing small-molecule drugs, and finds and verifies for the first time that W-5 can directly combine and inhibit NID1 protein.Starting from the core pathological link of "NID1-CSCs-resistance / metastasis", it is verified through experiments that W-5 hydrochloride can effectively combine with NID1, and can inhibit lung adenocarcinoma cell stemness, invasion and drug resistance by inhibiting the function of NID1 in lung adenocarcinoma.The application not only provides a clear orientation and a solid foundation for chemical optimization, structure modification, dosage form development and pharmacokinetics / toxicology (ADMET) research of a clear mechanism of directly targeting NID1, but also discovers new medicinal value of W-5 hydrochloride.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, specifically to a small molecule inhibitor targeting NID1 and its application. Background Technology

[0002] Lung adenocarcinoma is a leading cause of cancer-related death, with poor prognosis primarily attributed to delayed diagnosis, cancer stem cell (CSC)-mediated drug resistance, invasion, and metastasis. Nestin 1 (Nidogen-1, NID1), a key component of the basement membrane and extracellular matrix (ECM), has been shown to play a crucial role in tumor progression in various malignancies, including lung cancer. However, there are currently no commercially available small-molecule inhibitors targeting NID1.

[0003] Currently, biologics targeting NID1 that directly neutralize or induce an immune response include anti-NID1 monoclonal neutralizing antibodies and NID1 mRNA vaccines. Both NID1-targeting neutralizing antibodies and NID1-targeting mRNA vaccines are still in the research and development stage and are not yet commercially available, significantly hindering both basic research and clinical translation of NID1-targeted therapies. The clinical translation cycle for biologics is long, resulting in poor timeliness of application. Antibodies and mRNA vaccines are still in the basic research stage, requiring complex preclinical studies including process development, pharmacodynamics, pharmacokinetics, and safety assessments, as well as lengthy multi-phase clinical trials before final clinical application, failing to meet current urgent clinical needs. Existing NID1-targeting biologics have not fully elucidated the core role of NID1 in maintaining the stemness of cancer cells (CSCs), promoting drug resistance, and metastasis.

[0004] While existing anti-NID1 neutralizing antibodies can significantly inhibit lung cancer tumor growth, their effects are not ideal. The therapeutic potential of NID1-targeting anti-tumor mRNA vaccines for lung cancer is also under investigation, but their mechanism of action involves inducing a NID1-specific immune response to limit tumor progression. However, their mechanism of action does not directly target the NID1-mediated cancer cell resistance (CSC) characteristics, potentially limiting their ability to reverse NID1-mediated multidrug resistance and other NID1-mediated malignant phenotypes in lung cancer. Furthermore, current technologies are still in the research and development stage, facing long translational cycles and high costs. In recent years, drug repurposing has become an effective strategy. By applying known drugs to new therapeutic areas, drug development can be accelerated and costs reduced, providing a novel treatment strategy that can be rapidly translated into clinical practice. This strategy has shown great potential in the treatment of multidrug-resistant cancers. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide a small molecule inhibitor targeting NID1 and its application.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: the use of the prodrug of the compound shown in formula (I), its pharmaceutically acceptable salt, or its pharmaceutically acceptable solvate in the preparation of a drug targeting and inhibiting NID1, wherein the CAS number of the compound is 61714-25-8.

[0007] This invention screens existing small molecule drugs, using molecular docking prediction and functional verification, to identify W-5 hydrochloride as a small molecule that specifically inhibits the function and activity of NID1. Experimental verification shows that W-5 hydrochloride can effectively bind to NID1 and, in lung adenocarcinoma, inhibits NID1 function, thereby suppressing the stemness, invasion, and drug resistance of lung adenocarcinoma cells.

[0008] In a preferred embodiment of the application described in this invention, the targeted inhibition of NID1 is to inhibit the protein function and / or activity of NID1.

[0009] The present invention also provides the use of a prodrug of the compound shown in formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof in the preparation of a medicament for the prevention and / or treatment of lung adenocarcinoma, characterized in that the CAS number of the compound is 61714-25-8.

[0010] In a preferred embodiment of the application described in this invention, the drug inhibits the stemness of tumor stem cells by inhibiting NID1.

[0011] In a preferred embodiment of the application described in this invention, the drug reverses tumor cell drug resistance by inhibiting NID1.

[0012] In a preferred embodiment of the application described in this invention, the drug inhibits tumor cell proliferation, invasion, and metastasis by inhibiting NID1.

[0013] The present invention also provides a small molecule inhibitor targeting NID1, comprising a prodrug of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

[0014] The present invention also provides a method for screening NID1 inhibitors, wherein the method uses a prodrug of a compound represented by formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as a positive control or standard.

[0015] The present invention also provides a pharmaceutical composition for treating lung adenocarcinoma, comprising a prodrug of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

[0016] As a preferred embodiment of the pharmaceutical composition of the present invention, the pharmaceutical composition further includes an anticancer drug.

[0017] As a preferred embodiment of the pharmaceutical composition of the present invention, the anticancer drug includes cisplatin or osimertinib.

[0018] Preferably, the drug composition is administered via nebulization, intravenous injection, intraperitoneal injection, subcutaneous injection, intramuscular injection, or ocular administration.

[0019] The beneficial effects of the present invention: The present invention provides a small molecule inhibitor targeting NID1 and its application, which has the following advantages: (1) Novel mechanism of action and precise target: The present invention is the first to discover and verify that W-5 hydrochloride can directly bind to and inhibit NID1 protein (confirmed by SPR experiment, KD value reaches nM level). This reveals a completely new mechanism of action; (2) Targeting the core malignant phenotype of lung cancer: The present invention starts from the core pathological link of "NID1-CSCs-drug resistance / metastasis", and confirms that W-5 hydrochloride can significantly inhibit the NID1-mediated cancer stem cell phenotype from three dimensions: self-renewal ability (tumor spheroid formation), migration and invasion ability and drug resistance, thereby reversing the drug resistance of chemotherapy and targeted drugs from the root and overcoming the pain points of easy recurrence and easy metastasis of existing therapies; (3) Significant efficacy: The present invention confirms the significant efficacy of W-5 hydrochloride through in vitro cell proliferation inhibition, stem cell functional experiments, drug resistance reversal experiments and in vivo mouse xenograft models at multiple levels and in all aspects. It has anti-tumor activity and can effectively reverse the resistance of chemotherapy drugs (cisplatin) and targeted drugs (osimertinib) mediated by NID1 overexpression, and has shown good safety in vivo; (4) It has good prospects: Although W-5 hydrochloride is not a marketed drug, as a small molecule compound, it still has advantages such as relatively simple production process, controllable cost, good stability, easy storage and administration (potential oral possibility) compared to biological agents. Its clear mechanism of directly targeting NID1 provides a clear direction and solid foundation for its subsequent chemical optimization, structural modification, dosage form development and pharmacokinetic / toxicological (ADMET) studies, accelerating its progress towards clinical application. Attached Figure Description

[0020] Figure 1The results show the screening and molecular docking analysis of candidate compounds for NID1. A is a heatmap of the top 32 NID1 candidate inhibitor compounds screened using cMap, with color bars and blocks representing similarity scores; B is a comparison of NID1 expression levels using the COMPARE tool based on the NCI60 project; C is the GI50 value of W-5 in cancer cell lines, with the midline representing the mean NID1 expression or the mean log10 (GI50) value.

[0021] Figure 2 The image shows the 3D structure of NID1 obtained from protein homology modeling. The left side displays the NID1 drug-binding pocket. The right side shows the 2D structure of the drug, the active amino acid residues, molecular forces, and molecular spatial distances. The visualization analysis was performed using Discovery Studio software.

[0022] Figure 3 The affinity of W-5 hydrochloride for NID1 was determined using SPR. The figure shows the concentration-corrected binding kinetics curves. W-5 hydrochloride solutions were flowed at different concentrations (6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM, 200 μM) over the surface of a CM5 sensor chip immobilized with NID1. Different colored curves represent the corresponding concentration gradients. Experiments were conducted at 25°C in a pH 7.4 buffer system. Response values ​​are expressed in response units (RU).

[0023] Figure 4 W-5 hydrochloride was used to inhibit the uptake of CAF-EV-derived NID1 protein in A549 and HCC827 cells. A shows the expression of NID1 protein in A549 cells treated with CAF-EV-NID1; B shows the expression of NID1 protein in HCC827 cells treated with CAF-EV-NID1; β-Actin was used as an internal control for sample loading to verify the consistency and comparability of total protein loading amounts in each lane.

[0024] Figure 5 The study investigated the inhibitory effect of W-5 hydrochloride on the proliferation of lung cancer cells. A represents the effect of different concentrations of W-5 on the proliferation of A549 cells, detected using the MTT assay; B represents the effect of different concentrations of W-5 on the proliferation of HCC827 cells, detected using the MTT assay. Data are expressed as mean ± standard deviation, and statistical analysis was performed using Student's t-test.

[0025] Figure 6The stem cell characteristics of lung adenocarcinoma cells overexpressing NID1, inhibited by W-5, are shown in the following figures: A) bright-field image showing tumor spheroid formation (scale bar: 100 μm); B) quantitative statistical analysis of the number and diameter of tumor spheroids; C) image showing crystal violet-stained migrating cells (A549 cells scale bar: 100 μm; HCC827 cells scale bar: 200 μm); D) quantitative statistical analysis of the number of migrating cells; E) flow cytometry analysis of tumor stem cell markers; and F) quantitative statistical analysis of the ALDH⁺ / CD44⁺ cell ratio.

[0026] Figure 7 To illustrate W-5 reversal of CAF-EV-NID1-induced chemotherapy resistance, A represents the dose-response curves and IC50 values ​​of A459 cells treated with Cisplatin, and B represents the dose-response curves and IC50 values ​​of HCC827 cells treated with Osimertinib (*p<0.05, ****p<0.0001). Data are expressed as mean ± standard deviation, and statistical analysis was performed using Student's t-test.

[0027] Figure 8 Figure 1 shows the in vivo inhibition of NID1-mediated tumor growth by W-5. Figure 2 shows the experimental diagram of the LLC-1 cell C57BL / 6 mouse subcutaneous xenograft model. Figure 3 shows the tumor images of each group after day 18 of treatment. Figure 4 shows the statistical analysis of tumor weight (*p<0.05). Figure 5 shows the curve of mouse body weight change during treatment, where ns represents no significant difference. Figure 6 shows the dynamic curve of xenograft growth (*p<0.05, ****p<0.0001). All data are expressed as mean ± standard deviation (n=2). Statistical analysis was performed using one-way ANOVA and Tukey's post-hoc test. Detailed Implementation

[0028] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Unless otherwise specified, the various raw materials, reaction equipment, testing equipment, and testing methods used in the following embodiments are all known in the art.

[0029] Example 1 This embodiment experimentally verifies the binding site of the interaction between compound W-5 hydrochloride and NID1. The specific experimental steps are as follows: First, this study uses shRNA-mediated RNA interference technology to independently downregulate the expression of the NID1 gene. Two shRNA lentiviral vectors (shNID1-1 and shNID1-2) targeting different NID1 target sequences were designed and constructed, with non-target shRNA (shCtrl) as a control. The above lentiviral vectors were transduced into target cell lines, and after puromycin selection, stable NID1 knockdown cell lines were obtained. The knockdown efficiency of NID1 at the mRNA and protein levels was verified by real-time quantitative PCR (qRT-PCR) and Western blot technology. Subsequently, transcriptome sequencing (RNA-seq) analysis was performed on the shNID1-1, shNID1-2, and shCtrl cell lines to screen out differentially expressed genes common to both shNID1 and the control group, totaling 54 overlapping genes. Based on a comprehensive analysis of the cMap database, 10 potential small molecule compounds targeting NID1 were identified. The transcriptomic changes induced by these compounds were highly consistent with the gene expression alterations caused by NID1 knockdown. Further analysis using the NCI COMPARE tool to examine NID1 expression in pan-cancer cell lines and the corresponding GI50 concentrations of these compounds revealed that W-5 hydrochloride showed a good fit. Figure 1 ).

[0030] 3D structures of NID1 and the compound were obtained using AlphaFold 3.0 and the PubChem database. Homology modeling and molecular docking were performed using PyMol 2.5 and Pyrx 9.5. Finally, the docking pocket and interaction details were visualized using Biovia Discovery Studio Client 2020, suggesting that W-5 may be an effective NID1 inhibitor. The binding site of W-5 hydrochloride to NID1 is shown below. Figure 2 As shown.

[0031] To verify the direct binding interaction between W-5 hydrochloride and NID1, the interaction between W-5 hydrochloride and NID1 was monitored using surface plasmon resonance (SPR) technology with a BIAcore T200 (Cytiva) microarray. The experiment was conducted at 25°C in a low molecular weight, multi-cycle mode. Biotin-labeled NID1 protein was immobilized on the surface of a CM5 microarray (approximately 2000 RU). Subsequently, different concentrations (6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM, 200 μM) of W-5 hydrochloride were passed through the microarray, and real-time response curves (RU) were recorded. All experiments were performed in a pH 7.4 buffer system. The binding affinity (equilibrium dissociation constant KD value) for each interaction pair was calculated using the BIAcore T200 evaluation software (Cytiva). Didoxigenin was used as a negative control to exclude non-specific binding. Figure 3 As shown, the SPR results indicate that W-5 hydrochloride can directly bind to NID1, with a steady-state fitted KD value of 72.2 nM.

[0032] To further verify whether W-5 hydrochloride could block cellular uptake of NID1 protein, the specific experimental method was as follows: A549 and HCC827 lung cancer cells were treated with extracellular vesicles rich in NID1 protein derived from tumor-associated fibroblasts (CAFs) (CAF-EV-NID1), while simultaneously being treated with W-5 hydrochloride to detect whether W-5 hydrochloride could inhibit the binding of NID1 protein to tumor cells. Western blotting was used to detect NID1 protein expression in cells: after different treatments, total protein was extracted from cells, quantified using the BCA method, and equal amounts of protein samples were separated by SDS-PAGE electrophoresis and transferred to a PVDF membrane; after blocking with 5% BSA, anti-NID1 primary antibody (ab254325) was added and incubated overnight at 4°C. After washing with TBST, the cells were incubated with the corresponding HRP-labeled secondary antibody, and finally, the images were acquired using an ECL chemiluminescence system. The results are as follows: Figure 4 As shown, W-5 hydrochloride can downregulate NID1 protein expression in CAF-EV-NID1 treated tumor cells, indicating that W-5 hydrochloride may block the binding of extracellular NID1 to receptors on the surface of tumor cells, thereby inhibiting its function.

[0033] Example 2 This study investigated the inhibitory effect of W-5 hydrochloride on the phenotype of lung cancer cell proliferation (CSCs). Specifically, W-5 hydrochloride was used as a potential NID1 inhibitor, and its inhibitory effect on lung cancer cell proliferation was detected by the MTT assay. The specific experimental method is as follows: A549 and HCC827 cells in logarithmic growth phase were seeded into 96-well plates at a density of 5 × 10³ cells per well and pre-cultured at 37°C in a 5% CO2 incubator for 24 hours. After cell attachment, the medium was replaced with fresh medium containing different concentrations of W-5 hydrochloride (0 μM, 1.25 μM, 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM, 80 μM, 160 μM) and cultured for another 48 hours. 10 μL of MTT solution (5 mg / mL) was added to each well and incubated at 37°C for 4 hours. After terminating the culture, the supernatant was carefully aspirated, and 100 μL of dimethyl sulfoxide (DMSO) was added to each well. The plates were shaken for 10 minutes to fully dissolve the formazan crystals. The absorbance (OD value) of each well was measured at 490 nm using a microplate reader, and cell viability was calculated. The experimental results are as follows: Figure 5 As shown, W-5 inhibited the proliferation of two lung cancer cell lines in a concentration-dependent manner. In A549 cells, its IC50 value was 99.33 ± 5.81 μM; in HCC827 cells, the IC50 value was 50.78 ± 2.33 μM. To eliminate the interference of non-specific cytotoxicity on subsequent functional experiments, sublethal concentrations below the IC50 were selected in subsequent studies (A549: 50 μM; HCC827: 25 μM) to evaluate the specific effects of this inhibitor on tumor cell stemness and drug resistance.

[0034] Example 3 This study investigated the regulatory effect of W-5 hydrochloride on the stem cell characteristics of lung cancer. In a lung adenocarcinoma model overexpressing NID1, the intervention effect of W-5 hydrochloride was evaluated from three dimensions: self-renewal capacity, migration and invasion capacity, and stem cell marker expression. The specific experimental methods are as follows: In the tumor spheroid formation assay, single-cell suspensions were seeded into ultra-low adsorption 96-well plates (50 cells per well) and cultured for 14 days in serum-free stem cell conditioned medium (RPMI 1640 containing EGF, FGF, IGF, and B27). Medium was added every 3 days. After the assay, the number of spheroids with a diameter >50 μm was counted and the diameter of the spheroids was measured under an inverted microscope. In the Transwell migration assay, 200 μL of serum-free cell suspension (5 × 10⁶ cells / well) was added to the upper chamber of the Transwell. 4Cells were cultured in a 10% FBS medium at 800 μL in the lower chamber. After 48 hours of culture, cells that had migrated to the lower chamber were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and the cell count was performed by randomly selecting five fields of view under a microscope. For flow cytometry analysis, the treated cells were collected, and aldehyde dehydrogenase activity was detected using an ALDH assay kit (Stem Cell Technologies) following the manufacturer's instructions, co-incubated with anti-CD44-APC (BD Biosciences) antibody. The proportion of ALDH⁻ / CD44⁺ double-positive cells was detected using flow cytometry, and data analysis was performed using FlowJo software.

[0035] Experimental results are as follows Figure 6 As shown, W-5 hydrochloride treatment significantly inhibited the tumor spheroid formation ability of NID1-overexpressing cells. Figure 6 AB); Transwell assays showed that this compound effectively inhibited cell migration ( ). Figure 6 CD); Flow cytometry confirmed a significant decrease in the proportion of ALDH⁺ / CD44⁺ stem cell-like cells. Figure 6 The above results demonstrate, from three dimensions—self-renewal, migration capacity, and stem cell markers—that W-5 hydrochloride can inhibit the stemness of lung adenocarcinoma by targeting the NID1 pathway.

[0036] Example 4 This embodiment investigates the reversal effect of W-5 hydrochloride on NID1-mediated tumor drug resistance using the MTT assay. The specific experimental method is as follows: A549 and HCC827 cells were divided into three groups: a control group, a CAF-EV-NID1 treatment group, and a CAF-EV-NID1 and W-5 hydrochloride co-treatment group. After each group was treated for 24 hours, the cells were transferred to 96-well plates and cultured for 48 hours in fresh medium containing different concentrations of chemotherapeutic drugs (Cisplatin or Osimertinib). 10 μL of MTT solution (5 mg / mL) was added to each well, and the cells were incubated at 37°C for 4 hours. The culture was then terminated, the supernatant was discarded, and 100 μL of DMSO was added to dissolve formazan crystals. The absorbance was measured at 490 nm using a microplate reader. Cell viability was calculated, and dose-response curves were plotted. The IC50 value for each group was calculated using a Logistic model.

[0037] Experimental results are as follows Figure 7As shown, CAF-EV-NID1 treatment significantly enhanced the resistance of A549 and HCC827 cells to Cisplatin and Osimertinib, as evidenced by a significantly higher IC50 value compared to the control group. However, co-treatment with W-5 hydrochloride significantly reversed the CAF-EV-NID1-induced resistance effect, resulting in a significant decrease in the IC50 values ​​of both chemotherapeutic drugs. These results indicate that W-5 hydrochloride, targeting the NID1 pathway, can effectively inhibit acquired drug resistance in tumor cells.

[0038] Example 5 This embodiment investigates the inhibitory effect of W-5 hydrochloride on NID1-mediated lung cancer tumorigenesis. The specific experimental method is as follows: First, extracellular vesicles (CAF-EV-NID1) overexpressing NID1 from cancer-associated fibroblasts were prepared. The specific operation procedure is as follows: The full-length human NID1 cDNA was cloned into an expression vector, and then transfected into primary cultured human cancer-associated fibroblasts (CAFs). Stable NID1-overexpressing CAF cell lines were obtained through puromycin selection. The conditioned medium of this cell line was collected, and extracellular vesicles (EVs) were separated using differential ultracentrifugation. The specific steps included: first, centrifugation at 3,000×g for 20 minutes at 4°C to remove intact cells and large cell debris; then, the supernatant was collected and centrifuged at 10,000×g for 30 minutes at 4°C to remove organelles and large vesicles; finally, the supernatant was ultracentrifuged at 16,700×g for 4 hours at 4°C to obtain EV precipitate. The precipitate was detected by Western blot, and identified using EV markers such as CD63 and TSG101, as well as NID1 antibody. Protein concentration was determined using the BCA method, ultimately yielding CAF-EV-NID1. The control group CAF-EV (CAF-EV-Ctrl) was prepared using the same method from CAFs transfected with an empty vector. Subsequently, as... Figure 8As shown in Figure A, a subcutaneous xenograft tumor model of LLC-1 cells was constructed in C57BL / 6 mice. Mouse-derived LLC-1 lung cancer cells in the logarithmic growth phase were harvested, digested with trypsin, counted, and resuspended in serum-free culture medium. The cells were then mixed with an equal volume of pre-chilled matrix gel to prepare an inoculation suspension with a cell density of 2 × 10^5 cells / 100 μL (i.e., 50 μL of cell suspension mixed with 50 μL of matrix gel). 100 μL of the mixed suspension was drawn up using a pre-chilled syringe and subcutaneously injected into the right back of 6- to 8-week-old male C57BL / 6 mice. The health status and tumor growth of the mice were monitored daily during the experiment. Four days after tumor formation, the experimental group received NID1-overexpressing EV (tail vein injection, 20 μg / time, every 3 days) combined with W-5 hydrochloride (1 mg / kg, intraperitoneal injection every other day). After 18 days, W-5 hydrochloride showed significant inhibition of tumor growth without affecting mouse body weight. Figure 8 (BD), indicating no significant toxicity. Tumor growth curves showed that the tumor volume in the W-5 hydrochloride group was significantly smaller than that in the EV-NID1 group, indicating that W-5 hydrochloride can effectively inhibit the EV-NID1-mediated pro-tumorigenesis. Figure 8 E).

[0039] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. The use of the prodrug of the compound shown in formula (I), its pharmaceutically acceptable salt, or its pharmaceutically acceptable solvate in the preparation of a medicament targeting and inhibiting NID1, characterized in that, The CAS number of the compound is 61714-25-8 。 2. The application according to claim 1, characterized in that, The targeted inhibition of NID1 refers to the inhibition of the protein function and / or activity of NID1.

3. The use of the prodrug of the compound shown in formula (I), its pharmaceutically acceptable salt, or its pharmaceutically acceptable solvate in the preparation of a medicament for the prevention and / or treatment of lung adenocarcinoma, characterized in that, The CAS number of the compound is 61714-25-8 。 4. The application according to claim 3, characterized in that, The drug inhibits the stemness of tumor stem cells by inhibiting NID1.

5. The application according to claim 3, characterized in that, The drug reverses drug resistance in tumor cells by inhibiting NID1.

6. The application according to claim 3, characterized in that, The drug inhibits tumor cell proliferation, invasion, and metastasis by inhibiting NID1.

7. A small molecule inhibitor targeting NID1, characterized in that, This includes prodrugs of compounds represented by formula (I), pharmaceutically acceptable salts thereof, or pharmaceutically acceptable solvates thereof. 。 8. A pharmaceutical composition for treating lung adenocarcinoma, characterized in that, This includes prodrugs of compounds represented by formula (I), pharmaceutically acceptable salts thereof, or pharmaceutically acceptable solvates thereof. 。 9. The pharmaceutical composition according to claim 8, characterized in that, The pharmaceutical composition also includes an anticancer drug.

10. The pharmaceutical composition according to claim 9, characterized in that, The anticancer drugs mentioned include cisplatin or osimertinib.

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