A small molecule inhibitor targeting nlrc5 and applications thereof

Abstract

The application provides a small-molecule inhibitor targeting NLRC5 and an application, and an effective component of the small-molecule inhibitor is N5I-1 and a pharmaceutically acceptable salt thereof. The small-molecule inhibitor has blood-brain barrier permeability, can significantly inhibit the transcription function of brain nerve cells NLRC5 in an in-vivo environment, and further inhibits the expression of NLRC5 target genes MHC-I. Meanwhile, the small-molecule inhibitor has a significant inhibitory effect on the transcription of NLRC5 and the expression of MHC-I in various peripheral blood cells and immune cells. In monocyte / macrophage cells, brain glioma cells and lung cancer cells cultured in vitro, N5I-1 all exhibits a significant NLRC5 transcription inhibitory effect.

Description

Technical Field

[0001] This invention belongs to the field of biopharmaceutical technology, and in particular relates to a small molecule inhibitor targeting NLRC5 and its application. Background Technology

[0002] NLRC5 is a major transcriptional regulator of MHC-I molecules. Through an enhancer-dependent mechanism, it regulates the expression profile of MHC-I in immune cells, tissue cells, and blood cells, thereby directly regulating CD8. + The recognition and activation of antigens by T cells and NK cells influence adaptive immune surveillance and innate immune responses. For example, patent US20130177577A only proposes that NLRC5 can serve as a regulatory site for MHC-I expression. In 2024, Cell reported a novel role of NLRC5 in driving PANoptosis, further emphasizing the importance of this molecule in regulating immune inflammatory responses. However, due to the large protein structure (1915 Aa) and functional domains (101 Aa) of NLRC5, the regulation of NLRC5 expression can only be achieved through gene editing in experimental animals (CRISPR or conditional gene knockout), and specific small molecule inhibitors are still lacking. Summary of the Invention

[0003] In view of this, the present invention aims to overcome the deficiencies in the prior art and propose a small molecule inhibitor targeting NLRC5 and its application.

[0004] To achieve the above objectives, the technical solution of the present invention is implemented as follows: In a first aspect, the present invention provides the application of compound N5I-1 in the preparation of inhibitors targeting NLRC5, wherein the structural formula of compound N5I-1 is as follows: .

[0005] Secondly, this invention provides the application of compound N5I-1 in the preparation of antitumor drugs, wherein the structural formula of compound N5I-1 is: ; The tumor is either non-small cell lung cancer or glioma.

[0006] Thirdly, this invention provides the use of compound N5I-1 in the preparation of drugs for treating neuroinflammatory lesions, wherein the structural formula of compound N5I-1 is: .

[0007] Furthermore, the neuroinflammatory lesion is a neuroinflammatory lesion caused by chronic traumatic encephalopathy.

[0008] Fourthly, the present invention provides a pharmaceutical preparation for treating cancer, wherein the active ingredient of the pharmaceutical preparation is compound N5I-1 and its pharmaceutically acceptable salt, supplemented with pharmaceutically acceptable excipients.

[0009] The aforementioned pharmaceutical preparations can be formulated into liquid, solid, semi-solid, or gaseous formulations using pharmaceutically acceptable methods. Liquid dosage forms include solutions, injections, lotions, etc.; solid dosage forms include powders, pills, tablets, films, etc.; semi-solid dosage forms include ointments, gels, etc.; and gaseous formulations include aerosols, sprays, etc.

[0010] Compared with the prior art, the present invention has the following advantages: (1) The small molecule inhibitor described in this invention, namely compound N5I-1, has blood-brain barrier permeability, meaning it significantly inhibits the transcriptional function of NLRC5 in brain nerve cells. Simultaneously, it significantly inhibits the transcriptional activity of NLRC5 in various peripheral blood cells, immune cells, monocytes / macrophages, glioma cells, and lung cancer cells. This is manifested as a significant reduction in the transcriptional function of NLRC5 and a significant downregulation of the expression level of the NLRC5 target gene MHC-I.

[0011] (2) The small molecule inhibitor described in this invention, namely compound N5I-1, provides a new intervention method for brain injury diseases such as chronic traumatic encephalopathy and stroke, neuroimmunological diseases such as multiple sclerosis, and tumor diseases such as non-small cell lung cancer, glioma, and hepatocellular carcinoma. Attached Figure Description

[0012] Figure 1 Conformation analysis diagram of NLRC5 protein and N5I-1 / NLRC5 binding site; Figure 2 This is a diagram showing the contribution of each amino acid at the N5I-1 / NLRC5 binding site to molecular binding. Figure 3 The figures show the molecular dynamics simulations of N5I-1 and NLRC after docking (A is the root mean square deviation plot, B is the root mean square fluctuation plot, and C is the radius of gyration (Rg) plot). Figure 4 The interaction fingerprint of N5I-1 / NLRC5; Figure 5 The image shows the SPR (gold standard for molecular docking) test results for N5I-1 / NLRC5. Figure 6To construct an in vitro model of TNF-α+IFN-γ (T / I)-induced HT-22 neuron PANoptosis (Figure A shows the expression levels of NLRC5 and H-2K1 after N5I-1 intervention detected by RT-PCR, and Figure B shows the transcriptional activity of NLRC5 / H-2K1 after N5I-1 intervention detected by ChIP). Figure 7 IC50 curves of N5I-1 in human monocytic leukemia cell line THP-1 (A) and mouse monocytic macrophage leukemia cell line RAW264.7 (B); Figure 8 IC50 curves of N5I-1 in human non-small cell lung cancer cell line A549 (A) and human lung cancer cell line H292 (B); Figure 9 IC50 curves of N5I-1 in human glioblastoma cell line U87 (A) and human glioma cell line U251HT-22 (B); Figure 10 IC50 curves of N5I-1 in mouse microglia BV2 (A) and mouse hippocampal neuron cell line HT-22 (B); Figure 11 The first result of the N5I-1 in vivo intervention experiment (Figure A shows the purity of neurons, astrocytes, and microglia sorted from brain tissue of mice with chronic traumatic encephalopathy using flow cytometry (immunomagnetic bead method), and B and D show the expression levels of H-2K1 in neurons (B), astrocytes (C), and microglia (D) detected by RT-PCR, respectively). Figure 12 The second result of the N5I-1 in vivo intervention experiment (Figure A shows the neutrophils, monocytes, CD4+ T cells, CD8+ T cells and NK cells in the peripheral blood of mice with chronic traumatic encephalopathy, sorted by flow cytometry; B and F show the expression levels of H-2K1 in neutrophils (B), monocytes (C), CD4+ T cells (D), CD8+ T cells (E) and NK cells (F), respectively, detected by RT-PCR). Detailed Implementation

[0013] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0014] In this document, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0015] In this document, when values ​​are described as ranges, it should be understood that such disclosure includes disclosure of all possible subranges within that range, as well as the specific numerical values ​​falling within that range, regardless of whether the specific numerical value or specific subrange is explicitly specified.

[0016] In this article, the terms "multiple" or "more than" are used unless otherwise specified, referring to a quantity greater than or equal to 2. For example, "one or more" means one or more types.

[0017] In this document, the terms "preferred" and "more preferred" are used only to describe implementation methods or embodiments with better effects, and should be understood as not constituting a limitation on the scope of protection of this invention.

[0018] In this document, terms such as "further" are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of this invention.

[0019] In this article, the term "and / or" describes an association between objects, indicating that three relationships can exist. For example, A and / or B means: A or B, or A and B.

[0020] In this document, the term "about" means a specified value of + / - 10%, preferably + / - 5%, and more preferably + / - 1%.

[0021] In this article, the terms “include,” “including,” “have,” “contain,” etc., are all open-ended terms, meaning that they include but are not limited to.

[0022] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar to or equivalent to those described herein may be used in the implementation or testing of this invention.

[0023] This invention integrates AlphaFold3 protein conformation analysis, Schrödinger virtual screening, molecular docking, molecular dynamics simulation and other technologies, combined with surface plasmon resonance (SPR) detection and in vitro and in vivo verification experiments, to successfully develop the NLRC5 small molecule inhibitor N5I-1, providing a reliable tool for achieving in vivo targeted inhibition of NLRC5.

[0024] The present invention will be described in detail below with reference to the embodiments.

[0025] The chemical name of N5I-1 in the following examples is: N-propyl-2,4-bis(4-methylbenzyl)-5-oxo-2,3,3a,4-tetrahydro-5H-oxazolo[3,2-a]pyrimidine-3-carboxamide, and its structural formula is: Its chemical structure can also be represented by the following SMILES: CCCNC(C(C(c1cc(NC(c2ccc(C)cc2)=O)ccc1)O1)N(Cc2ccc(C)cc2)C1=O)=O, purchased from ChemDiv, CA USA, with the number HIT103776906.

[0026] Example 1: Screening and molecular simulation of N5I-1, a small molecule inhibitor targeting and inhibiting NLRC5 1.1 Screening of small molecule inhibitors targeting NLRC5 (1) Experimental methods ① NLRC5 Target Structure Prediction and Pocket Identification: The structure of the NLRC5 protein (UniProt C3VPR6) was predicted using the AlphaFold3 server, and the NACHT-LRR core region containing candidate binding pockets was used as the acceptor input for subsequent pocket identification, docking, and molecular dynamics simulations. The prediction model was repaired with missing heavy atoms using the PDBFixer tool in the OpenMM suite and hydrogenated at pH 7.4. The orientation of polar hydrogens was then optimized using the Reduce program to establish a reasonable hydrogen bond network, which served as the acceptor input file for subsequent pocket identification and docking calculations. Candidate binding pockets were identified using POCASA (Roll algorithm): the full-atom model was discretized with a 1.0 Å grid, and a rolling probe with a radius of 1.2 Å was used to scan the protein surface. Open grooves were first filtered using the PSP (Protein Surrounded Points) rule, and then candidate pockets were ranked using a Volume-Depth comprehensive score. The pocket with the highest score was selected as the search box center for subsequent ChemDiv library docking. This pocket is located in the inner groove between NLRC5 LRR and NACHT.

[0027] ② ChemDiv Compound Library Preparation and Drug-likeness Screening: The starting library for virtual screening was the ChemDiv compound set. Preprocessing was first performed using RDKit: SMILES normalization and desalting (retaining only the principal component), generating a single three-dimensional conformation using the ETKDGv3 algorithm, and performing 200-step energy minimization with an MMFF94 force field; protonated states were estimated using Open Babel at pH 7.4 and hydrogenated; drug-likeness filtering simultaneously used the Lipinski five rules (MW ≤ 500, LogP ≤ 5, HBD ≤ 5, HBA ≤ 10) and the Veber rules (rotation bond ≤ 10, TPSA ≤ 140 Å). 2Based on the PAINS (Pan-Assay Interference Compounds) reactive substructure table, potential interfering molecules (aldehydes, Michael acceptors, α-halocarbonyl groups, etc.) are eliminated, and the screened molecules enter the docking stage.

[0028] ③ Molecular docking and MM-GBSA (Molecular Mechanics Generalized Born Surface Area) re-evaluation: Molecular docking was performed using AutoDock Vina 1.2. Receptors and ligands were converted to PDBQT format using Meeko. The search box was centered on the pocket center given by POCASA, with a size of approximately 24 × 24 × 24 Å. 3 A three-round hierarchical docking strategy is adopted to balance computational overhead and sampling accuracy: the first round uses extremely low precision (exhaustiveness = 1) to sample the entire ChemDiv database (approximately 1.6 × 10⁻⁶). 6 The initial screening process involved rapid evaluation of molecules, with the top 10% based on Vina scores advancing to the second round. The second round used standard precision (exhaustiveness = 10) to re-interact the initial results, with the top 10% again advancing to the third round based on Vina scores. The third round used ultra-high precision (exhaustiveness = 100) for the most refined inter-merging and scoring. The top 10% of compounds from the third round underwent single-frame MM-GBSA re-evaluation using gmx_MMPBSA in GB-Neck2 implicit solvent (igb=5). Further analysis of Vina docking scores, MM-GBSA binding energies, and the plausibility of interaction modes with key residues (Val516, Tyr240, Thr413, His517, etc.) was used to screen candidate compounds for subsequent activity testing. The top 10 MM-GBSA compounds were used for plotting and analysis. HIT103776906 was ultimately selected and named N5I-1 (Vina total score -9.4 kcal / mol, MM-GBSA single-frame ΔG_bind = -81.64 kcal / mol) as the research object for subsequent 100 ns all-atom molecular dynamics verification.

[0029] (2) Experimental results The results are as follows Figure 1 and Figure 2 As shown, it can be seen that N5I-1 and NLRC5 are perfectly connected at the three levels of global, local, and two-dimensional structure; the combination mode of the two is mainly hydrophobic contact, accompanied by hydrogen bonding and π-type interactions.

[0030] 1.2 Molecular simulation of small molecule inhibitors targeting NLRC5 (1) Experimental methods Molecular dynamics simulations were performed in GROMACS 2024.03. The protein was modeled using the Amber ff14SB force field; N5I-1 was parameterized using GAFF; the AM1-BCC charge was calculated using the antechamber module of AmberTools; and the water model was TIP3P. The system was placed in a cubic periodic cell with the solute at least 1.0 nm from the cell wall, and 0.15 mol / L NaCl was added to neutralize the net charge to simulate physiological ionic strength. Energy minimization was performed using the steepest descent method at 5 × 10⁻⁶. 4 Step to maximum force < 1000 kJ·mol -1 ·nm -1 Subsequently, equilibration was performed at 100 ps NVT (v-rescale, temperature controlled to 300 K) and 100 ps NPT (Parrinello-Rahman, pressure controlled to 1 bar) under positional constraints. Finally, the constraints were released for a 100 ns NPT simulation, with one frame output every 10 ps. Long-range electrostatics were processed using Particle-Mesh Ewald, truncated to 1.0 nm; all H bonds were constrained by LINCS, with an integration step size of 2 fs. RMSD, RMSF, and Rg were calculated using GROMACS' built-in tools based on the entire trajectory (protein fitting, ligand statistics in the fitted reference frame). Residue-ligand interaction fingerprints (hydrogen bonds, hydrophobic contacts, π-cations, etc.) were statistically analyzed frame by frame within 9000-9990 frames (the last 1000 frames, corresponding to 90-100 ns) using PLIP (Protein-Ligand Interaction Profiler). The cumulative frequencies of residues were summarized and the contact relationships between residues and ligand atoms were displayed in a Sankey diagram.

[0031] (2) Experimental results The results are as follows Figure 3 and Figure 4 As shown, it can be seen that: Figure 3The root mean square deviation (RMSD) of A reflects whether the binding system has reached equilibrium in molecular dynamics simulations. If the RMSD of the protein backbone fluctuates slightly within a certain range, it indicates that the molecular dynamics simulation of the protein-small molecule complex system has reached equilibrium. The RMSD curves of the NLRC5 backbone, ligand N5I-1, and binding site rapidly increase within 0-20 ns and then enter a relative plateau phase. No obvious overall drift is observed between the curves, indicating that the NLRC5 / N5I-1 complex is in a stable state. The density distribution on the right side mainly shows a single peak, with only a slight secondary peak shown by the ligand, suggesting that there is no significant overall drift or large-scale conformational change in the trajectory. These results indicate that the NLRC5 / N5I-1 binding system is stable. Figure 3 The root mean square fluctuation (RMSF) of B is used to analyze the degrees of freedom of movement of amino acid residues, i.e., flexibility. The RMSF of the main residues of NLRC5 falls between 0.5 and 1.5 Å, with an average of 1.14 Å, indicating that the overall structural flexibility of the protein molecule is low (most <2.0 Å), and the RMSF near the N5I-1 binding site is also small, indicating that the binding region is relatively stable; Figure 3 The radius of gyration (Rg) of C reflects the overall tightness of the protein and ligand. The Rg curves and three-component curves of NLRC5 and N5I-1 show relatively small fluctuations, indicating that the NLRC5 / N5I-1 complex did not show overall loosening or collapse within 100 ns; Figure 4 The residue-ligand interaction fingerprint shows that the steady-state binding of N5I-1 to the NLRC5 pocket depends on two hydrogen bonds (Val516-Lig:N3, Tyr240-Lig:O4), a set of hydrophobic contacts mainly composed of Val / Leu / Ala / Pro / Phe / Thr, and a low-frequency Arg263 Pi-Cation contact.

[0032] Example 2 Molecular docking experiment of N5I-1 and NLRC5 (1) Experimental methods The binding curves of NLRC5 with different concentrations of N5I-1 were determined using surface plasmon resonance (SPR) experiments (the gold standard for molecular interaction detection). The purified recombinant protein was immobilized on a CM5 sensor chip, and the binding affinity between N5I-1 and NLRC5 was subsequently determined using a GE Healthcare Biacore S200 molecular interaction instrument.

[0033] (2) Experimental results The results are as follows Figure 5 As shown, N5I-1 and NLRC5 can stably bind to form a complex. Single-concentration experiments show that N5I-1 has high binding capacity with NLRC5 at concentrations of 62.5 μM, 125 μM, 250 μM, 500 μM, and 1000 μM.

[0034] Example 3: In vitro activity test of N5I-1 3.1 Inhibitory effect of N5I-1 on the transcriptional function of NLRC5 / MHC-I (1) Experimental methods A TNF-α+IFN-γ-induced panapoptosis model of HT-22 neurons was constructed in vitro (20 ng / ml TNF-α + 10 ng / ml IFN-γ, treatment for 36 h) to activate the NLRC5 / H-2K1 (mouse MHC-I) transcriptional complex. The expression levels of NLRC5 and H-2K1 and the transcriptional activity of NLRC5 / H-2K1 after N5I-1 intervention were detected by RT-PCR and ChIP-PCR, respectively. The RT-PCR primers for H-2K1 were: forward primer, 5'-GAGACACAGGTCGCCAAGAAC-3' (as shown in SEQ ID NO.1); reverse primer, 5'-CGCTGGTAAGTGTGAGAGCC-3' (as shown in SEQ ID NO.2). The ChIP-PCR primers are: forward primer, 5'-CCGCGGACGCTGGATA-3' (as shown in SEQ ID NO.3); reverse primer, 5'-GGCGATTCGCGACTTCTG-3' (as shown in SEQ ID NO.4).

[0035] (2) Experimental results The results are as follows Figure 6 As shown, it can be seen that: Figure 6 A shows that after inducing panapleosis in in vitro neurons, intervention with N5I-1 can significantly reduce the expression level of H-2K1 mRNA (without affecting the expression of N5I-1 mRNA). Figure 6 B shows that after intervention with N5I-1, the binding of NLRC5 to the H2K1 promoter was effectively inhibited. Both results indicate that N5I-1 has an inhibitory effect on the transcriptional function of NLRC5 / MHC-I.

[0036] 3.2 Inhibitory effect of N5I-1 on transcriptional function of human and mouse mononuclear / macrophage cell lines (1) Experimental methods Human THP-1 monocyte cell line and mouse monocyte / macrophage RAW 264.7 cell line expressing NLRC5 and MHC-I molecules under physiological conditions were selected as the research subjects. Cells were treated with N5I-1 for 24 h. The expression levels of HLA-B (human HLA-I) and H-2K1 after intervention with different concentrations of N5I-1 were detected by RT-PCR. The RT-PCR primers for HLA-B were: forward primer, 5'-CAGTTCGTGAGGTTCGACAG-3' (as shown in SEQ ID NO.5); reverse primer, 5'-CAGCCGTACATGCTCTGGA-3' (as shown in SEQ ID NO.6).

[0037] (2) Experimental results The results are as follows Figure 7 A- Figure 7 As shown in Figure B, IC50 < 100, indicating that N5I-1 has a significant inhibitory effect on the transcription of MHC-I in both monocytes and macrophages.

[0038] 3.3 Inhibitory effect of N5I-1 on transcriptional function of lung cancer cells (A549 and H292) (1) Experimental methods Human lung cancer cell lines A549 and H292, which express NLRC5 and MHC-I molecules under physiological conditions, were selected as the research subjects. Cells were treated with N5I-1 for 24 h. The expression level of HLA-B after intervention with different concentrations of N5I-1 was detected by RT-PCR.

[0039] (2) Experimental results The results are as follows Figure 8 A- Figure 8 As shown in Figure B, IC50 < 100, indicating that N5I-1 has a significant inhibitory effect on the transcription of HLA-B in both types of lung cancer cells.

[0040] 3.4 Inhibitory effect of N5I-1 on transcriptional function of glioma cells (U87 and U251) (1) Experimental methods Human glioma cell lines U87 and U251, which express NLRC5 and MHC-I molecules under physiological conditions, were selected as the research subjects. Cells were treated with N5I-1 for 24 h. HLA-B expression levels after intervention with different concentrations of N5I-1 were detected by RT-PCR.

[0041] (2) Experimental results The results are as follows Figure 9 A- Figure 9 As shown in Figure B, IC50 < 100, indicating that N5I-1 has a significant inhibitory effect on the transcription of HLA-B in two types of glioma cells.

[0042] 3.5 Inhibitory effect of N5I-1 on transcriptional function of BV2 and HT-22 cells (1) Experimental methods The mouse microglia BV2 and neuronal cell line HT-22, which expressed low levels of NLRC5 and MHC-I molecules under physiological conditions, were selected as research subjects. A TNF-α+IFN-γ (T / I)-induced PANoptosis in vitro model was constructed to activate the NLRC5 / H-2K1 transcriptional complex. Cells were treated with N5I-1 for 24 h. RT-PCR was used to detect the expression level of H-2K1 after intervention with different concentrations of N5I-1.

[0043] (2) Experimental results The results are as follows Figure 10 A- Figure 10 As shown in Figure B, IC50 < 100, indicating that N5I-1 has a significant inhibitory effect on the transcription of H-2K1 in microglia and neurons.

[0044] Example 4: In vivo intervention experiment of N5I-1 (1) Experimental methods 1) A mouse model of chronic traumatic encephalopathy (CTE) inducing central nervous system and systemic chronic inflammation was prepared. 100 μL of N5I-1 at a concentration of 40 μg / mL (80 μM) was injected via the tail vein 72 h after injury, and subsequent experiments were carried out at 6, 12 and 24 h thereafter.

[0045] 2) Neurons, astrocytes, and microglia from the brain tissue of CTE mice were sorted using magnetic immunomagnetic bead sorting (MACS). The purity of the sorted cells was assessed by flow cytometry, and the expression level of MHC class I molecule H-2K1 in these cells was detected by RT-PCR. Furthermore, neutrophils, monocytes, and CD4+ cells from the peripheral blood of CTE mice were sorted using flow cytometry (FACS). + T cells, CD8 + T cells and NK cells were analyzed, and the expression level of H-2K1 in these cells was detected by RT-PCR.

[0046] 3) The reagent kit and antibody information used in the previous experiment are as follows: ① Neuron isolation kit (#130-115-390; Miltenyi Biotec, Bergisch Gladbach, Germany), anti-GLAST magnetic beads (#130-095-826; Miltenyi Biotec), anti-CD45 magnetic beads (#130-052-301; Miltenyi Biotec), and anti-CD11b magnetic beads (#130-093-636; Miltenyi Biotec).

[0047] ② All flow cytometry antibodies were purchased from BioLegend, San Diego, CA, USA; including: APC / Cy7-CD11b (#101226), PerCP / Cy5.5-CD45 (#103132), APC-Ly6C (#128016), PE-Ly6G (#127608), PE / Cy7-CD3 (#100220), APC-CD4 (#100516), PE-CD8 (#100708), PerCP / Cy5.5-CD19 (#115534), and Biotin-NK1.1 (#108704).

[0048] (2) Experimental results The results are as follows Figure 11 and Figure 12 As shown, it can be seen that: Figure 11 A indicates that flow cytometry was used to identify and confirm that the three types of nerve cells obtained by MACS sorting had high purity. Figure 11 B- Figure 11 D indicates that N5I-1 can significantly inhibit the expression of H-2K1 in neurons, astrocytes, and microglia, demonstrating its ability to effectively cross the blood-brain barrier. Figure 12 A demonstrates a gating strategy for separating various blood cells and immune cells from peripheral blood using flow cytometry sorting technology; Figure 12 B- Figure 12 F indicates that N5I-1 significantly inhibited the activity of peripheral blood neutrophils, monocytes, and CD4+. + T cells, CD8 + H-2K1 expression in T cells and NK cells.

[0049] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The application of compound N5I-1 in the preparation of inhibitors targeting NLRC5, characterized in that: The structural formula of the compound N5I-1 is: 。 2. The application according to claim 1, characterized in that: The application of compound N5I-1 in the preparation of antitumor drugs, wherein the tumor is non-small cell lung cancer or glioma.

3. The application according to claim 1, characterized in that: Application of compound N5I-1 in the preparation of drugs for treating neuroinflammatory lesions.

4. The application according to claim 3, characterized in that: The neuroinflammatory lesions mentioned are neuroinflammatory lesions caused by chronic traumatic encephalopathy.

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