Application of VARS1 gene, VARS1 inhibitor and application

Abstract

The invention belongs to the field of medicines, and discloses application of VARS1 protein in screening and preparing prostatic cancer (PCa) treatment medicament development and application of a kit for diagnosing prostatic cancer and prostatic cancer malignant progression. According to the application, the effect of the VARS1 in the PCa is defined, the medicine which is specifically combined with the VARS1 in a targeted manner is screened, the killing effect of the medicine on the PCa cells is verified, and a new treatment thought and medicine are provided for further improving the PCa treatment effect. Meanwhile, the invention also discloses a plurality of compounds screened based on the gene, and experiments prove the effects of the compounds on the cellular level and the animal level. In addition, the invention also provides related applications of the compounds.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more particularly to the application of the VARS1 gene and protein, VARS1 inhibitors, and their uses. Background Technology

[0002] Prostate cancer (PCa) is one of the leading causes of death among middle-aged and elderly men. For early-stage PCa patients, androgen deprivation therapy (ADT) is a widely used first-line treatment because it blocks the androgen / androgen receptor (AR) signaling pathway essential for PCa cell survival. ADT is highly effective in the initial stages of treatment, but with prolonged treatment, almost all patients relapse within two years into castration-resistant prostate cancer (CRPC), often accompanied by metastasis. Currently, CRPC is not only incurable but also has a high mortality rate; this is mainly because the molecular mechanisms of its malignant progression are not yet fully understood, leading to a lack of therapeutic targets and very limited practical treatment options. Therefore, it is necessary to strengthen research on the mechanisms of CRPC development and progression, assess its high-risk factors, and identify new therapeutic targets or strategies. Summary of the Invention

[0003] The purpose of this invention is to provide an application of the VARS1 gene and protein. This invention clarifies the role of VARS1 in PCa, screens drugs that specifically target and bind to VARS1, and verifies their in vitro and in vivo killing effects on PCa cells, providing new treatment ideas and drugs for further improving the treatment effect of PCa.

[0004] Furthermore, this invention also discloses several compounds obtained based on screening of this protein, and experimentally demonstrates the effects of these compounds at the cellular and animal levels; in addition, this invention provides related applications of these compounds.

[0005] To achieve the above objectives, this application discloses:

[0006] Applications of VARS1 protein or VARS1 gene in screening and developing drugs for the treatment of prostate cancer, and in developing kits for diagnosing prostate cancer and its progression.

[0007] Valine tRNA synthetase 1 (VARS1) participates in valine tRNA biogenesis and encodes the only known valine cytoplasmic aminoacyl-tRNA synthetase. The protein encoded by the VARS1 gene belongs to the class I amino acid tRNA synthetase (aaRS) family. VARS1 catalyzes the high-fidelity binding of valine to homologous tRNA, a process known as valine charging. aaRS plays a driving or supporting role in tumor development. Mutations, aberrant expression, uncontrolled secretion, and interactions with other oncogenic factors can all affect intracellular aaRS expression levels, leading to imbalances in the metabolome and proteome, and ultimately inducing cancer. Furthermore, aaRS abnormalities support cancer growth by promoting protein synthesis and cell proliferation. Changes in aaRS, both at the gene and protein levels, ultimately affect protein synthesis and metabolome homeostasis (particularly the metabolome related to amino acids, energy, and RNA). Currently reported diseases associated with VARS1 (mainly its mutations) include neurodevelopmental disorders with microcephaly, epilepsy, cortical atrophy, and oxidative phosphorylation deficiency, but its application in the treatment of prostate cancer has not yet been observed.

[0008] To verify the feasibility of VARS1 as a therapeutic target, this invention employed computer-generated virtual screening of the reported VARS1 AF-P26640-F1-v4 structure. Polar hydrogen atoms and charges were added to protein molecules using Autodock Tools 1.5.6, and the molecules were then converted to PDBQT format. The SPECS database was split using Openbabel software, and the compounds were batch-converted to PDBQT format using a script. Batch molecular docking was performed using Autovina 1.1.2. Virtual scoring based on binding energies identified hundreds of small molecule compounds that might interact with VARS1. These small molecule compounds were screened from over one million compounds, and after verification, three compounds were found to significantly inhibit the proliferation of prostate cancer cells.

[0009] Then, through in vitro experiments, small molecule inhibitors that can bind to and inhibit VARS1 activity were further screened, and it was confirmed that they target and bind to VARS1 in prostate cancer cells and inhibit the proliferation of prostate cancer cells.

[0010] Meanwhile, in this invention, VARS1 expression increased sequentially in normal prostate tissue, prostate intraepithelial neoplasia, prostate carcinoma in situ, and castration-resistant prostate cancer. Downregulating VARS1 expression in prostate cancer cell lines significantly inhibited cell proliferation, migration, and stemness. This result indicates that VARS1 plays a role as an oncogene in prostate cancer. Following the general research approach in this field, detecting VARS1 expression holds promise for diagnosing the malignant progression of prostate cancer.

[0011] In specific embodiments, the expression of VARS1 increased sequentially in normal prostate tissue, prostate intraepithelial neoplasia, prostate carcinoma in situ, and castration-resistant prostate cancer.

[0012] In a preferred embodiment, the diagnostic reagent includes an immunohistochemical staining reagent targeting VARS1 protein expression.

[0013] Therefore, the application of the VARS1 gene proposed in this invention in the screening and development of drugs for the treatment of prostate cancer can be the application of substances in the form of small molecule compounds, peptides, protein degradation targeted chimeras (PROTACs) derived from small molecules, and molecular glutes that can specifically bind to the VARS1 protein as drugs for the treatment of prostate cancer. Alternatively, it can be the application of various optional detection methods in the field, such as primer-based, primer-probe-based, modified antibody-based, antibody-mediated immunohistochemical staining methods, and other related detection reagents and kits that can clearly characterize the gene and its expression level.

[0014] Based on this, the present invention also discloses a VARS1 inhibitor, wherein the VARS1 inhibitor is one or more compounds represented by Formula 1 to Formula 3 below;

[0015]

[0016] Formula 1

[0017]

[0018] Formula 2

[0019]

[0020] Formula 3

[0021] R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R 20 R 21 R 22 R 23 R 24 R 25 R 26 R 27 R28 R 29 R 30 Each is independently selected from H, OH, CX3, CHX2, CH2X, OCX3, OCHX2, OCH2X, CN, C(O)Y1, C(O)OY1, C(O)NY1Y2, C(O)NHY1, NHC(O)Y1, NY1C(O)Y2, NY1C(O)OY2, NY1OY2, N3, NY1C(O)Y2, NY1NY2Y3, SO4Y1, SO3Y1, SO2Y1, SO4NY1Y2, SO3NY1Y2, SO2NY1Y2 or SF5;

[0022] Where X is H, Cl, F, Br or I; Y1, Y2, Y3 are each independently selected from OH, CCl, CBr3, CF3, CI3, CH2Cl, CH2F, CH2Br, CH2I, CHCl2, CHF2, CHBr2, CHI2, CN, NH2, COOH, CONH, NO2, SH, SO3H, SO2HN2, SO4H, HNNH2, ONH2, NHC(O) NHNH2, NHC(O) NH2, NSO2H, NHC(O) H, NHC(O) OH, NHOH, OCCI3, OCBr3, OCF3, OCI3, OCHCl2, OCHBr2, OCHF2, OCHI2.

[0023] In the above-mentioned VARS1 inhibitors, the VARS1 inhibitor is one or more compounds shown in Formulas 4 to 6 below;

[0024]

[0025] Formula 4

[0026]

[0027] Formula 5

[0028]

[0029] Formula 6.

[0030] The chemical formulas corresponding to Equations 4 to 6 above are numbered ZINC04656109, ZINC02211211, and ZINC08443122 in the database, respectively. In some of the figures below, ZINC04656109, ZINC02211211, and ZINC08443122 are referred to as numbers 2701, 2901, and 3201. All subsequent experiments of this invention were purchased from the holder of this database and conducted accordingly.

[0031] In addition, the present invention also discloses the use of the VARS1 inhibitor described above in the preparation of anticancer drugs.

[0032] In the above-mentioned uses, the target of the anticancer drug is the VARS1 gene.

[0033] Of the above uses, the applicable condition for the anticancer drug is prostate cancer.

[0034] Finally, the present invention also discloses an anticancer drug containing the VARS1 inhibitor as described above, wherein the anticancer drug is an anticancer drug for treating prostate cancer.

[0035] This application has at least the following beneficial effects:

[0036] 1. This invention clarifies the role of VARS1 in PCa, screens drugs that specifically target and bind to VARS1 and verifies their killing effect on PCa cells, providing new treatment ideas and drugs for further improving the detection and treatment efficacy of PCa;

[0037] 2. The research results of this invention clarify the role and mechanism of ZINC04656109 (No. 2701), ZINC02211211 (No. 2901) and ZINC08443122 (No. 3201) in prostate cancer. According to the research results of this invention, inhibiting VARS1 can achieve a therapeutic effect on prostate cancer and inhibit the growth of prostate tumors.

[0038] 3. The results of this invention screened drugs that specifically target and bind to VARS1, which is beneficial for the development of clinical treatment drugs for prostate cancer and has important clinical significance. Attached Figure Description

[0039] Figure 1 The expression of VARS1 in normal prostate tissue, prostate intraepithelial neoplasia, prostate carcinoma in situ, and castration-resistant prostate cancer as described in Example 1.

[0040] Figure 2 This describes the cell biological function following downregulation of VARS1 expression in the PCa cell line as described in Example 1; wherein, Figure 2 A represents the verification of VARS1 expression after downregulation. Figure 2 B represents cell proliferation. Figure 2 C stands for stem cell, Figure 2 D stands for cell migration. Figure 2 E represents a xenograft subcutaneous tumor.

[0041] Figure 3 This is a schematic diagram of the VARS1 fraction inhibitor screening process.

[0042] Figure 4A The chemical structural diagrams of compounds ZINC04656109, ZINC02211211 and ZINC08443122 are shown.

[0043] Figure 4B The structural formulas of compounds ZINC00629069 and ZINC08435127 and their interaction curves with VARS1 are shown.

[0044] Figure 5 The graphs show the interactions between ZINC04656109, ZINC02211211 and ZINC08443122 and VARS1 as confirmed by in vitro fluorescence titration experiments.

[0045] Figure 6 The effects of ZINC04656109, ZINC02211211, and ZINC08443122 on the proliferation of prostate cancer cells.

[0046] Figure 7 The effects of compounds ZINC04656109, ZINC02211211 and ZINC08443122 on the proliferation of VARS1 knockdown cells and control cells are shown.

[0047] Figure 8A This is a plasmid map of the pLKO.1-EGFP-Puro lentiviral vector.

[0048] Figure 8B The effects of three small molecule compounds on cell proliferation were analyzed in VARS1 knockdown cells and control cells.

[0049] Specific implementation methods

[0050] The present invention will now be clearly and completely described in conjunction with embodiments thereof. It should be noted that, unless otherwise specified in the embodiments, conditions are performed under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products. Unless otherwise specified, all parts used in the embodiments of the present invention are parts by weight.

[0051] Example 1: Analysis of the role of the VARS1 gene in prostate cancer

[0052] The role and mechanism of VARS1 in prostate cancer remain unclear. Based on prostate cancer cell lines, we have confirmed that VARS1 plays the role of an oncogene in prostate cancer and identified its potential as a therapeutic target for prostate cancer.

[0053] 1.1 Expression of VARS1 in prostate cancer patients

[0054] To clarify the expression of VARS1 in prostate cancer patients, this study collected clinical samples from 13 CRPC patients, 10 PCa patients, and 10 normal prostate tissue samples. Immunohistochemical staining was used to detect the expression level of VARS1 protein.

[0055] The immunohistochemical staining procedure includes:

[0056] Sample processing: dewaxing and hydration (paraffin sections) or fixation (frozen sections / cell smears).

[0057] Antigen retrieval: thermal retrieval (citrate buffer) or enzyme retrieval.

[0058] Blocking: Treat with 3% H2O2 for 10 minutes (HRP system).

[0059] Sealing: 5% BSA, room temperature sealing for 30 minutes.

[0060] Antibody incubation: Primary antibody overnight at 4°C / 1-2 h at room temperature HRP / fluorescence (Proteintech, 15931-1-AP), secondary antibody 30-60 min at room temperature.

[0061] Color development: DAB color development or direct observation of fluorescence.

[0062] Counterstaining and mounting: counterstain with hematoxylin, and mount with neutral resin.

[0063] Test results are as follows Figure 1 The results show that the expression of VARS1 protein increased sequentially in normal tissue, PIN, PCa and CRPC, indicating that the protein expression level of VARS1 increases with the malignancy of PCa.

[0064] 1.2 Effects of downregulating VARS1 expression on cell biological function in prostate cancer cell lines

[0065] In this embodiment, siRNA was used to downregulate VARS1 expression in two prostate cancer cell lines, PC3 and LNCaP. Figure 2 A) siRNA inhibits protein expression by specifically degrading target mRNA through RNA interference mechanisms.

[0066] The specific procedure is as follows: Seed cells in 6-well plates. When the confluence reaches 60-70%, dilute 5 μl of Lipofectamine 2000 and 50-100 nM siRNA in 250 μl of Opti-MEM, respectively. After standing at room temperature for 5 minutes, mix to form a liposome-siRNA complex, and continue to stand for 15 minutes. Discard the cell culture medium and wash with PBS. Add 500 μl of Opti-MEM medium and transfection complex, and incubate at 37°C for 6 hours. Then replace with complete culture medium and continue to incubate for 24-72 hours before detecting protein expression levels.

[0067] This method utilizes liposomes to encapsulate siRNA, promoting its entry into cells. Subsequently, the siRNA binds to the RISC complex and guides the degradation of target mRNA, thereby achieving gene silencing.

[0068] The siRNA sequence used is as follows:

[0069] siNC: 5'-UUCUCCGAACGUGUCACGUTT-3';

[0070] siVARS1#542: 5'-GUUACGCCGACACGGAGUUAATT-3';

[0071] siVARS1#1782: 5'-CCUCGUGUCCUUUGCCUAUAATT-3';

[0072] siVARS1#3313: 5'-GUCUACUUGGAGUGCCUGAAATT-3'.

[0073] Finally, the effects on cell biological functions were verified using cell counting experiments, cell colony formation experiments, cell spheroidization experiments, and transwell experiments.

[0074] The results showed that downregulating VARS1 expression in prostate cancer cell lines could inhibit cell proliferation. Figure 2 B), cell cloning ( Figure 2 C), cell spheroidization ( Figure 2 D), and cell migration ( Figure 2 E).

[0075] Example 2: Drugs that specifically target and bind to VARS1

[0076] Since high expression of VARS1 is highly correlated with the occurrence and development of prostate cancer, novel targeted drugs can be designed based on VARS1. This embodiment first uses the AF-P26640-F1-v4 structure of VARS1 for computer-generated virtual screening to identify small molecule compounds that may interact with VARS1. Then, in vitro experiments are used to further screen small molecule inhibitors that can bind to and inhibit VARS1 activity, confirming their targeted binding to VARS1 within prostate cancer cells and their inhibition of prostate cancer cell proliferation.

[0077] 2.1 Computer-generated virtual screening revealed that ZINC04656109, ZINC02211211, and ZINC08443122 interact with VARS1.

[0078] This invention uses the SPECS database (306,709 small molecule compounds, http: / / www.specs.net) for virtual screening based on molecular docking. The human VARS1 (AF-P26640-F1-v4) protein structure database is from AlphaFold2 (https: / / alphafold.com / ). Polar hydrogen atoms and charges were added to the protein molecules using Autodock Tools 1.5.6, and the molecules were finally converted to PDBQT format. To further determine the binding pockets between the small molecules and the protein compounds, we first predicted the protein binding pockets using the proteinplus website (https: / / proteins.plus / ). The AutoDockTools software was used to generate a parameter file for the docking box based on the predicted pocket locations. Molecular docking was performed using AutoDock Vina software. PyMOL was used to search for hydrogen bond interactions. A two-dimensional interaction map was obtained from the proteinplus website.

[0079] The coordinates of the active pocket of the VARS1 protein molecule were set as follows: center_x = 10.137, center_y = 1.818, center_z = -11.350, size_x = 114, size_y = 72, size_z = 68, spacing = 1.000, exhaustiveness = 32, num_modes = 10. Unless otherwise specified, all other parameters used their default values. Finally, the SPECS database was split using Openbabel software, then batch-converted to PDBQT format using a script, and then batch molecular docking was performed using Autodock vina 1.1.2. Virtual evaluation and scoring were performed based on the binding energy. The screening process is as follows: Figure 3Through high-throughput virtual screening, we ultimately identified 42 candidate inhibitors of VARS1. To evaluate the cytotoxicity of these inhibitors to PCa cells, we purchased 35 compounds from the SPECs compound library (as some compounds were out of stock). Subsequently, we screened these compounds using an MTT assay.

[0080] The results are as follows Figure 4A As shown, 16 compounds were found to have significant inhibitory effects on PC3, DU145 and LNCaP in PCa cells.

[0081] 2.2 In vitro confirmation of the targeted binding of small molecule compounds ZINC04656109, ZINC02211211 and ZINC08443122 to VARS1

[0082] The screening results in section 4.1 were evaluated based on the diversity of chemical structures and binding energies. Sixteen small molecule compounds were selected, and the interaction between the small molecule compounds and VARS1 was further confirmed using fluorescence titration.

[0083] The specific method is as follows: Prepare a 100mM compound stock solution, take 1.0μL of the stock solution and add it to 99 μL of buffer (consistent with VARS1 protein buffer), and dilute to 1000μM; add 100 μL of VARS1 protein solution (5 μM) to a 96-well plate, and add an equal volume of buffer to the parallel control.

[0084] Then, a fixed volume of compound solution was added (0, 1, 2, 4, 6, 8, 10, and 20 μL when the compound bound to VARS1), and the mixture was gently pipetted and incubated at 4°C for 10 minutes. Fluorescence values ​​were read for each well under excitation at 280 nm (332 nm). The fluorescence values ​​of the corresponding buffer wells were subtracted from the protein well fluorescence values ​​(to remove optical interference from the compound), and the decrease in fluorescence intensity with increasing compound amount was observed to conform to a dose-response relationship.

[0085] The results are as follows Figure 5 As shown, the fluorescence titration results indicate that ZINC04656109, ZINC02211211, and ZINC08443122 can bind to VARS1.

[0086] refer to Figure 4B ZINC00629069 and ZINC08435127, as controls, did not target and bind to VARS1 in vitro (Kd>10μM).

[0087] 2.3 Effects of compounds ZINC04656109, ZINC02211211 and ZINC08443122 on the biological function of prostate cancer cells

[0088] 2.3.1 Inhibition of cell proliferation

[0089] Based on the results of fluorescence titration, this embodiment further evaluates the effects of three small molecule compounds on cell proliferation in the PC3 cell line;

[0090] Cells were seeded into 96-well plates and then treated with a VARS1 candidate inhibitor. Cell viability was assessed using the MTT assay. Absolute viability measurements were normalized to the DMSO control group and expressed as percentage viability, as shown below. Figure 6 As shown, ZINC04656109, ZINC02211211 and ZINC08443122 all inhibited the proliferation of PC3 and LNCaP cells.

[0091] 2.3.2 Mouse Experiment

[0092] The effect of this drug on the in vivo growth of PCa was examined using a subcutaneous mouse tumor-bearing model. Specifically, 50 μl of PBS cell suspension and 50 μl of matrix gel were mixed and injected subcutaneously into the cells. When the tumor size reached 70 mm... 3 Around 100 μL / 20g body weight, mice were divided into groups based on tumor size. The drug was prepared as a mixed solution of 10% DMSO, 20% PEG300, 5% Tween80, and 65% ddH2O. Mice (BALB / c-nu) were injected intraperitoneally with this solution at a dose of 100 μL / 20g body weight, every 2-3 days for a total of 8-10 injections, after which they were sacrificed for tissue collection. The dosages for each injection were: 2701: 15 mg / kg, 2901: 10 mg / kg, 20 mg / kg; 3201: 10 mg / kg, 20 mg / kg.

[0093] The results are as follows Figure 7 As shown, ZINC04656109, ZINC02211211 and ZINC08443122 were found to inhibit the progression of prostate cancer in vivo;

[0094] Figure 7 In the diagram, 2701 represents ZINC04656109, 2901 represents ZINC02211211, and 3201 represents ZINC08443122.

[0095] 2.3.3 Cell knockdown experiment

[0096] Based on this, this embodiment analyzes the effects of three small molecule compounds on cell proliferation in VARS1 knockdown cells and control cells;

[0097] The specific method is as follows: using the pLKO.1-EGFP-Puro lentiviral vector (see map) Figure 8A A VARS1 knockdown plasmid was constructed, and a lentiviral vector was generated through homologous recombination. Subsequently, three plasmids (lentiviral plasmid, psPax2, and pMD2.G) were co-transfected into HEK293T cells with polyethylenimine and packaged into lentiviral particles.

[0098] The method for constructing lentiviral plasmids is as follows: Homologous sequences of 15-50 bp are designed at both ends of the target fragment and the linearized pLKO.1-EGFP-Puro lentiviral vector, and precise splicing is achieved using recombinase; the steps include:

[0099] 1) When amplifying the target fragment by PCR, add a homologous arm to the 5' end of the primer;

[0100] The sequence of the target fragment is:

[0101] shVARS1#542:

[0102] 5'ccggGTTACGCCGACACGGAGTTAACTCGAGTTAACTCCGTGTCGGCGTAACTTTTT 3'; (SEQID NO.1)

[0103] shVARS1#3313:

[0104] 5'ccggGTCTACTTGGAGTGCCTGAAACTCGAGTTTCAGGCACTCCAAGTAGACTTTTT 3'. (SEQ ID NO.2)

[0105] The primers used for vector construction are:

[0106] shVARS1#542-Forward: tggaaaggacgaaacaccggtCCGGGGAGGATTTCATCTCTTGTATCTCGAGATACAAG; (SEQ ID NO.3)

[0107] shVARS1#542-Reverse: AagttatgtaacgcggaattcAAAAAGGAGGATTTCATCTCTTGTATCTCGAGATACAA; (SEQ ID NO.4)

[0108] shVARS1#3313-Forward:

[0109] tggaaaggacgaaacaccggtccggGTCTACTTGGAGTGCCTGAAACTCGAGTTTCAGG; (SEQ IDNO.5)

[0110] shVARS1#3313-Reverse:

[0111] AagttatgtaacgcggaattcaaaaaGTCTACTTGGAGTGCCTGAAACTCGAGTTTCAG; (SEQ IDNO.6)

[0112] 2) Simultaneously prepare linearized vectors with homologous ends (enzyme sites EcoRI and AgeRI);

[0113] 3) Mix the fragment and vector in the specified ratio, and incubate with recombinase;

[0114] 4) Directly transform competent cells, screen for positive clones, and verify them through sequencing;

[0115] 5) Extract and preserve plasmids.

[0116] Transfection mixtures were prepared in OPTI-MEM solution at a molar ratio of 10:7.5:3.5. Viral supernatants were harvested at 48 and 72 hours post-transfection. Viral infection was performed at room temperature in the presence of polyebrene. Stable vars1 knockdown or overexpression PCa cell lines were obtained via lentiviral transduction and then screened with puromycin (1 μg / mL) for 2–4 days. At this point, the stably knockdown VARS1 cell lines were successfully constructed. The effect of VARS1 candidate inhibitors on the viability of stably knockdown VARS1 prostate cancer cell lines was further evaluated using the MTT assay. The resistance of stably knockdown VARS1 cell lines to treatment with VARS1 candidate inhibitors was also compared with the control group.

[0117] The results are as follows Figure 8B As shown, cell lines with knocked-down VARS1 did indeed exhibit enhanced resistance to VARS1 candidate inhibitors. Therefore, at the cellular level, the targeting and inhibition of VARS1 by three small molecule compounds were confirmed.

[0118] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description; thus, all changes falling within the meaning and scope of the claims are intended to be embraced within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

Claims

1. Use of VARS1 protein in screening and preparing a medicament for developing a drug for treating prostate cancer.

2. Use of VARS1 gene or VARS1 protein in developing a kit for diagnosing prostate cancer and malignant progression of prostate cancer.

3. A VARS1 inhibitor, characterized in that, The VARS1 inhibitor is a compound represented by one or more of the following Formula 1 to Formula 3; R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 21 , R 22 , R 23 , R 24 , R 25 , R 26 , R 27 , R 28 , R 29 , R 30 each is independently selected from H, OH, CX3, CHX2, CH2X, OCX3, OCHX2, OCH2X, CN, C(O)Y1, C(O)OY1, C(O)NY1Y2, C(O)NHY1, NHC(O)Y1, NY1C(O)Y2, NY1C(O)OY2, NY1OY2, N3, NY1C(O)Y2, NY1NY2Y3, SO4Y1, SO3Y1, SO2Y1, SO4NY1Y2, SO3NY1Y2, SO2NY1Y2, or SF5; wherein X is H, Cl, F, Br or I; Y1, Y2, Y3 are each independently selected from OH, CCl, CBr3, CF3, CI3, CH2Cl, CH2F, CH2Br, CH2I, CHCl2, CHF2, CHBr2, CHI2, CN, NH2, COOH, CONH, NO2, SH, SO3H, SO2HN2, SO4H, HNNH2, ONH2, NHC(O)NHNH2, NHC(O)NH2, NSO2H, NHC(O)H, NHC(O)OH, NHOH, OCCI3, OCBr3, OCF3, OCI3, OCHCl2, OCHBr2, OCHF2, OCHI2.

4. The VARSI inhibitor of claim 3, wherein, The VARS1 inhibitor is a compound represented by one or more of the following Formula 4 to Formula 6; 5. Use of the VARS1 inhibitor according to claim 3 or 4 for preparing an anticancer drug.

6. Use according to claim 5, characterized in that, The target of the anticancer drug is VARS1 protein.

7. Use according to claim 5, characterized in that, The anticancer drug is used for treating prostate cancer.

8. An anticancer drug, characterized in that, The anticancer drug contains the VARS1 inhibitor according to claim 3 or 4.

9. The anticancer drug according to claim 8, characterized by, The anticancer drug is an anticancer drug for treating prostate cancer.

Images