Compound containing 2, 4-quinazolinedione and application of compound in antitumor drugs

Abstract

The invention discloses a compound containing 2, 4-quinazolinedione and application of the compound in antitumor drugs, and relates to the technical field of antitumor drugs. The 2, 4-quinazolinedione-containing skeleton is taken as a core, the whole shows a stronger and more stable inhibition trend to EGFR targets, and the advantages of target ends can be better conducted to tumor cell proliferation inhibition, so that the positioning of efficient EGFR inhibitors / efficient anti-tumor drugs is supported. Compared with the limitation that an existing EGFR inhibitor is insufficient in mutation subtype selectivity, drug resistance is easily generated after long-term medication, and the curative effect is attenuated, the EGFR inhibitor wholly points to the trend that the EGFR inhibitor has more advantages on a drug resistance related model, and the drug resistance is higher. Aiming at the common problems of pharmacokinetic defects and clinical application limitation caused by off-target toxicity in the prior art, through the design thought of novel structure and performance optimization, the invention integrally presents the trend of'more hopeful consideration of curative effect and safety and better druggability '.

Description

Technical Field

[0001] This invention relates to the field of antitumor drug technology, specifically to a compound containing 2,4-quinazolinedione and its application in antitumor drugs. Background Technology

[0002] Cancer, as a malignant disease that seriously threatens human health worldwide, continues to rise in incidence and mortality rates, making clinical treatment extremely urgent. Traditional anti-tumor methods such as chemotherapy and radiotherapy can inhibit tumor growth to a certain extent, but due to the lack of target specificity, they are prone to causing irreversible damage to normal cells, leading to serious side effects such as bone marrow suppression and gastrointestinal reactions. Moreover, their efficacy against advanced metastatic tumors is limited, making it difficult to meet the clinical demand for safe and effective treatment options.

[0003] With the development of precision medicine technology, targeted anti-tumor drugs have become a core direction in anti-tumor drug development because they can specifically act on key driving targets of tumor cells and significantly reduce toxicity to normal tissues. Among them, epidermal growth factor receptor (EGFR), as an important tumor-related target, is overexpressed or activatingly mutated in various malignant tumors such as lung cancer, breast cancer, and colorectal cancer. By continuously activating signaling pathways such as cell proliferation and inhibiting apoptosis, it directly promotes the occurrence and progression of tumors. Therefore, EGFR inhibitors have become a research focus in the field of targeted therapy.

[0004] While currently marketed EGFR inhibitors have achieved some clinical efficacy, they still have many limitations: some inhibitors are not selective enough for EGFR mutation subtypes (such as the T790M resistance mutation), resulting in poor efficacy; long-term use can easily induce new drug resistance mechanisms in tumor cells, causing the drug to quickly become ineffective; at the same time, some compounds have pharmacokinetic defects (such as low bioavailability and excessively rapid metabolism in vivo) or off-target toxicity problems, which further limit their clinical application.

[0005] 2,4-Quinazolidinedione compounds, due to their unique heterocyclic structure, can efficiently bind to kinase active sites through hydrogen bonding and hydrophobic interactions, demonstrating excellent potential in the development of kinase inhibitors. Studies have confirmed that these scaffold compounds exert antitumor effects by inhibiting EGFR activity. However, existing EGFR inhibitors containing 2,4-quinazolidinedione structures still require breakthroughs in terms of activity potency, target selectivity, resistance overcoming ability, and safety, failing to fully meet the clinical demand for highly effective and low-toxicity antitumor drugs. Therefore, developing novel and optimized EGFR inhibitors containing 2,4-quinazolidinediones has significant clinical value and practical implications for advancing antitumor drug development and improving patient prognosis. Summary of the Invention

[0006] The purpose of this invention is to address the problems existing in the prior art by providing a compound containing 2,4-quinazolinidone and its application in antitumor drugs. It can serve as a highly effective EGFR epidermal growth factor receptor inhibitor, suitable for use in the active pharmaceutical ingredient field, and can also be used to prepare highly effective and low-toxicity antitumor drugs, thereby overcoming the limitations of existing antitumor drugs and meeting clinical treatment needs.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is: a compound containing 2,4-quinazolinidone, wherein the compound has the structure shown in Formula 1:

[0008] Formula 1: ;

[0009] The application of the compound or its tautomers or pharmaceutically acceptable salts or solvates in the field of active pharmaceutical ingredients;

[0010] R1 in Formula 1 is a substituent;

[0011] R1 is any one of hydrogen, halogen, nitro, alkyl with 1-10 carbon atoms, alkoxy with 1-10 carbon atoms, or ester with 2-10 carbon atoms;

[0012] Or R1 may be an alkyl group with 1-10 carbon atoms substituted by a halogen.

[0013] Furthermore, the halogen is selected from F or Cl.

[0014] Furthermore, the alkyl group having 1-10 carbon atoms is selected from any one of methyl, ethyl, propyl, isopropyl, and tert-butyl.

[0015] Furthermore, the alkoxy group having 1-10 carbon atoms is selected from any one of methoxy, ethoxy, and propoxy.

[0016] Furthermore, the ester group with 2-10 carbon atoms is selected from ester groups with 2-10 carbon atoms, such as propyl ester group and ethyl ester group.

[0017] Furthermore, the alkyl group with 1-10 carbon atoms substituted by the halogen is selected from trifluoromethyl.

[0018] Furthermore, the compound is selected from any one of the compounds shown in the following structures:

[0019] ;

[0020] ; .

[0021] Application of a compound containing 2,4-quinazolinidone in the field of protein inhibitors.

[0022] Furthermore, the protein inhibitor is an EGFR epidermal growth factor receptor inhibitor.

[0023] Application of a compound containing 2,4-quinazolinidone in the preparation of antitumor drugs.

[0024] A compound containing 2,4-quinazolinedione is a chemical pharmaceutical raw material and can be used in the manufacture of formulations containing chemical pharmaceutical raw materials.

[0025] Furthermore, the tumor is one or more of the following: non-small cell lung cancer (NSCLC), colorectal cancer, head and neck squamous cell carcinoma (HNSCC), breast cancer, pancreatic cancer, and gastric cancer.

[0026] Compared with the prior art, the beneficial effects of the present invention are:

[0027] 1. This invention uses a 2,4-quinazolinedione skeleton as its core, and as a whole, it exhibits a stronger and more stable inhibitory trend on the EGFR target. It can also better transmit the target advantage to the inhibition of tumor cell proliferation, thereby supporting the positioning of "highly effective EGFR inhibitor / highly effective anti-tumor drug".

[0028] 2. Compared with the limitations of existing EGFR inhibitors, such as insufficient selectivity for mutant subtypes and the tendency to develop drug resistance and thus reduce efficacy with long-term use, this invention generally points to the trend of "being more advantageous for drug resistance-related models and having stronger resistance to drug resistance".

[0029] 3. In response to the common pharmacokinetic defects and off-target toxicity that limit clinical application in existing technologies, this invention presents a trend of "more promising to balance efficacy and safety, and better drug-likeness" through a novel structural and performance-optimized design approach. Attached Figure Description

[0030] Figure 1 This is the synthetic route for a compound containing 2,4-quinazolinedione as described in this invention.

[0031] Figure 2 This is a molecular docking diagram of compound 1 described in this invention with the EGFR epidermal growth factor receptor. Detailed Implementation

[0032] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are merely some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] Example 1

[0034] Synthesis of Compound 1:

[0035] ;

[0036] Step 1-1:

[0037] Compound 1-a: 3-bromo-4-fluoro-N,N-dimethylaniline;

[0038] Compound 1-b: 4-ethynylphenol;

[0039] Compound 1-c: 3-(4-ethynylphenoxy)-4-fluoro-N,N-dimethylaniline;

[0040] In a dry, clean, round-bottom flask equipped with a stirrer, add 5.00 g of compound 1-a, 3.25 g of compound 1-b, 0.43 g of CuI, 0.64 g of N,N-dimethylglycine hydrochloride, and 14.94 g of cesium carbonate. Connect the flask to a vacuum / nitrogen purging system, evacuate the system, and backfill with nitrogen. Repeat this operation three times to ensure the reaction system is under inert gas protection. Under nitrogen protection, add 70 mL of anhydrous 1,4-dioxane via a syringe. Place the flask in an oil bath preheated to 100°C, start vigorous stirring, and reflux for 24 hours. After the reaction is complete, cool the reaction solution to room temperature. Filter the reaction solution using diatomaceous earth to remove insoluble inorganic salts, and wash the filter cake thoroughly with ethyl acetate (50 mL × 3). Combine the filtrate and washings, transfer them to a separatory funnel, and wash successively with water (100 mL) and saturated saline (100 mL). After organic phase separation, the product was dried with anhydrous sodium sulfate for 30 minutes; the desiccant was removed by filtration, and the filtrate was concentrated by rotary evaporation under reduced pressure to obtain the crude product; the crude product was purified by rapid silica gel column chromatography with an elution gradient of n-hexane / ethyl acetate = 20:1 to 10:1. The fraction containing the target product was collected, the solvent was removed by rotary evaporation, and the product was dried under vacuum to obtain 4.18 g of compound 1-c.

[0041] Step 2-1

[0042] Compound 1-c: 3-(4-ethynylphenoxy)-4-fluoro-N,N-dimethylaniline;

[0043] Compound 2-a: 3-(tert-butyl)-6-chloroquinazoline-2,4(1H,3H)-dione;

[0044] Compound 1: 3-(tert-butyl)-6-((4-(5-(dimethylamino)-2-fluorophenoxy)phenyl)vinyl)quinazolin-2,4(1H,3H)-dione;

[0045] In a dry, clean, round-bottom flask equipped with a stirrer, add 4.18 g of compound 1-c, 4.95 g of compound 2-a, 0.57 g of Pd(PPh3)2Cl2, and 0.31 g of CuI. Evacuate the mixture using a double-row tube and backfill with nitrogen (or argon) gas, repeating this process three times to ensure the reaction system is under inert gas protection. Under nitrogen protection, add 60 ml of anhydrous DMF and 4.97 g of triethylamine. Start stirring and place the reaction mixture in an oil bath preheated to 90°C. The mixture was heated to reflux and stirred for 16 hours. After the reaction, the reaction solution was cooled to room temperature. The reaction mixture was filtered through a funnel lined with diatomaceous earth, and the filter cake was washed with a small amount of ethyl acetate. The filtrate was transferred to a separatory funnel, diluted with ethyl acetate (100 mL), and then washed with water (3 × 50 mL) and saturated brine (50 mL), respectively. The organic phase was dried over anhydrous sodium sulfate for 30 minutes and then filtered. The filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography. Gradient elution was used with a mobile phase of n-hexane / ethyl acetate (gradually changing from 10:1 to 2:1). The fraction containing the target product was collected, concentrated under reduced pressure, and dried under vacuum to obtain 5.61 g of compound 1.

[0046] Structural assessment:

[0047] Compound 1-c, mass spectrometry m / z-M+1: 256; NMR. 1 HNMR-CDCl3:

[0048] δ7.55-7.45(m,2H),7.09-6.96(m,3H),6.61(m,1H),6.40(dd,1H),3.09(s,1H),2.86(s,6H);

[0049] Compound 1, mass spectrometry m / z-M+1: 472; NMR. 1 HNMR-CDCl3:

[0050] δ8.07(d,1H),7.61-7.41(m,4H),7.12-6.93(m,3H),6.61(m,1H),6.40(dd,1H),2.86(s,6H),1.51(s,9H).

[0051] Examples 2-10

[0052] In Examples 2-10, compounds 2-10 were prepared sequentially, following the preparation method of Example 1, except that compounds 1-b were replaced, and the rest remained the same as in Example 1. For details, please refer to Table 1.

[0053] Table 1

[0054]

[0055]

[0056] Comparative compound 1: 3-(4-chloro-3-phenoxybenzyl)-7-fluoroquinazoline-2,4(1H,3H)-dione;

[0057] Comparative compound 2: Erlotinib;

[0058] Comparative compound 3: Dacomitinib.

[0059] Performance testing:

[0060] 1. EGFR epidermal growth factor receptor IC 50 :

[0061] The target compounds and control compounds (erlotinib, dacomitinib, and control compound 1) prepared in Examples 1-10 were dissolved in DMSO to prepare a 10 mM stock solution, which was then serially diluted with kinase buffer to eight concentration gradients (10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM, and 0.003 μM) to ensure that the final DMSO concentration in each well was ≤1%. In a 384-well microplate, 2 μL of the serially diluted compound solution and 4 μL of recombinant EGFR kinase solution were added sequentially. 2 μL of peptide substrate solution (final concentration 5 nM) and 20 μM of peptide substrate solution were added and incubated at room temperature for 10 min to allow the compound to fully bind with the enzyme. 2 μL of ATP solution (final concentration 10 μM, matching the ATP Km value) was added, quickly mixed, and incubated at 37°C for 60 min. 10 μL of the stop solution from the LFRET assay kit (containing a phosphatase inhibitor and fluorescent probe) was added, and the mixture was incubated at room temperature in the dark for 20 min. The fluorescence signal values ​​of each well were then detected using a microplate reader (Ex = 485 nm, Em = 520 nm).

[0062] Negative and positive controls: Negative control (no compound group): kinase buffer was used instead of the compound solution; Positive control (complete inhibition group): a known EGFR kinase inhibitor was added; Blank control: kinase buffer was used instead of all reaction components;

[0063] Calculate the EGFR kinase inhibition rate at each compound concentration: Inhibition rate (%) = [1 - (sample well fluorescence value - blank control value) / (negative control value - blank control value)] × 100%, and fit the IC50 value. 50 The data is shown in Table 2.

[0064] 2. Non-small cell lung cancer (NSCLC) IC 50 :

[0065] H1975 cells were placed in culture flasks containing complete culture medium and cultured at 37°C in a 5% CO2 incubator until the logarithmic growth phase. Cells were then collected by digestion with 0.25% trypsin, washed twice with PBS, and resuspended in complete culture medium to adjust the cell concentration to 5 × 10⁶ cells / mL. 4 Cells were seeded at a density of 100 μL / well in a 96-well plate and incubated at 37°C for 24 h to allow cell adhesion. The original culture medium was then discarded, and 100 μL of complete culture medium containing different concentrations of the compound (10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM, 0.003 μM) was added to each well, with six replicates for each concentration. A negative control (complete culture medium containing 1% DMSO) and a blank control (complete culture medium only, cell-free) were also included. The 96-well plate was incubated at 37°C in a 5% CO2 incubator for 72 h. 10 μL of CCK-8 reagent was added to each well, gently vortexed, and incubated at 37°C for 2 h. The absorbance (OD) of each well was then measured using a microplate reader. 450 Inhibition rate (%) = [1 - (sample well OD value - blank control OD value) / (negative control OD value - blank control OD value)] × 100%, and the IC50 value is fitted. 50 The data is shown in Table 2.

[0066] Table 2

[0067] compound source <![CDATA[EGFR epidermal growth factor receptor IC 50 (nM)]]> <![CDATA[H1975(NSCLC)IC 50 (µM)]]> Example 1 3.2 0.18 Example 2 6.5 0.32 Example 3 4.8 0.25 Example 4 2.1 0.12 Example 5 9.7 0.55 Example 6 15.4 0.92 Example 7 7.9 0.41 Example 8 28 1.8 Example 9 12.2 0.75 Example 10 18.6 1.1 Comparative compound 1 45 3.6 Comparative compound 2 2.6 12 Comparative compound 3 6 1.9

[0068] The compounds in the examples generally exhibited strong and similar inhibitory activities at the EGFR enzyme activity level, indicating that this series binds stably to the target. Simultaneously, they also demonstrated good overall inhibitory effects on H1975 cells, with cell activity and enzyme activity generally changing in the same direction, suggesting that target inhibition can be effectively transmitted to the cell phenotype. However, a few compounds showed a "good enzyme activity, but only average cell activity" or the opposite, reflecting the significant contribution of ADME factors such as cell permeability, efflux, and metabolic stability to the cellular level. Among the comparative compounds, the first-generation EGFR inhibitors showed significantly weaker performance on H1975 cells, consistent with the drug resistance background of this cell model. While the irreversible / broader-spectrum controls were relatively more advantageous at the cellular level, they were not necessarily superior to the best examples in all aspects. The overall trend supports the potential for further optimization and screening of leading compounds in this series for drug resistance-related cell models.

[0069] 3. Molecular docking: Compound 1 underwent molecular docking with the EGFR epidermal growth factor receptor, with the following binding sites: LYS692, LEU694, GLY695, ALA698, PHE699, VAL702, LYS704, ALA719, LYS721, LEU723, ALA731, GLU734, ILE735, GLU738, MET742, CYS751, LEU764, TH R766, GLN767, LEU768, MET769, PRO770, PHE771, GLY772, CYS773, ASP776, TYR777, GLU780, HIS7 81. ASP813, ARG817, ASN818, LEU820, THR830, ASP831, GLY833, LEU834, LYS851, VAL852, PRO853.

[0070] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A compound containing 2,4-quinazolinedione, characterized in that, The compound has the structure shown in Formula 1: Formula 1: ; The application of the compound or its tautomers or pharmaceutically acceptable salts or solvates in the field of active pharmaceutical ingredients; R1 in Formula 1 is a substituent; R1 is any one of hydrogen, halogen, nitro, alkyl with 1-10 carbon atoms, alkoxy with 1-10 carbon atoms, or ester with 2-10 carbon atoms; Or R1 may be an alkyl group with 1-10 carbon atoms substituted by a halogen.

2. A compound containing 2,4-quinazolinidone according to claim 1, characterized in that, The halogen is selected from F or Cl.

3. A compound containing 2,4-quinazolinidone according to claim 1, characterized in that, The alkyl group having 1-10 carbon atoms is selected from any one of methyl, ethyl, propyl, isopropyl, and tert-butyl.

4. A compound containing 2,4-quinazolinidone according to claim 1, characterized in that, The alkoxy group having 1-10 carbon atoms is selected from any one of methoxy, ethoxy, and propoxy.

5. A compound containing 2,4-quinazolinidone according to claim 1, characterized in that, The ester group with 2-10 carbon atoms is selected from ester groups with 2-10 carbon atoms, such as propyl ester group and ethyl ester group.

6. A compound containing 2,4-quinazolinidone according to claim 1, characterized in that, The alkyl group with 1-10 carbon atoms substituted by the halogen is selected from trifluoromethyl.

7. A compound containing 2,4-quinazolinidone according to claim 1, characterized in that, The compound is selected from any one of the compounds shown in the following structures: ； ； 。 8. The use of a compound containing 2,4-quinazolinidone as described in any one of claims 1-7 in the field of protein inhibitors.

9. The application of a compound containing 2,4-quinazolinidone according to claim 8 in the field of protein inhibitors, characterized in that, The protein inhibitor is an EGFR epidermal growth factor receptor inhibitor.

10. The use of a compound containing 2,4-quinazolinidone as described in any one of claims 1-7 in the preparation of an antitumor drug.

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