An acylhydrazine compound with antitumor activity, its preparation method and application

Abstract

This invention relates to the field of biomedical technology, specifically to an acylhydrazine compound with antitumor activity, its preparation method, and its applications. In this invention, an acylhydrazine compound was prepared that can act as a USP16 inhibitor, exhibiting highly efficient activity in inhibiting the proliferation of breast cancer cells, providing a safe and effective candidate drug molecule for the treatment of breast cancer. The synthesis method of this acylhydrazine compound is simple, the raw materials are readily available, and it has good functional group compatibility. Experimental results show that the tumor inhibition rate of this acylhydrazine compound in the tumor-bearing mouse group is superior to that in the positive control group, demonstrating promising application prospects.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to an acylhydrazine compound with antitumor activity, its preparation method, and its application. Background Technology

[0002] Breast cancer is a malignant tumor that seriously threatens women's health worldwide. It exhibits extremely high heterogeneity and can be classified into molecular subtypes such as Luminal A, Luminal B, HER2-overexpressing breast cancer, and triple-negative breast cancer (TNBC) based on the expression status of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). Among these, TNBC, lacking targets for ER, PR, and HER2, is insensitive to endocrine therapy and HER2-targeted therapy, resulting in a high recurrence rate and poor prognosis, making it a challenging area for clinical treatment. Therefore, a deeper understanding of the molecular mechanisms underlying breast cancer development and progression, and the discovery of new therapeutic targets and biomarkers, are crucial for improving patient outcomes.

[0003] Ubiquitin-specific protease 16 (USP16), a member of the deubiquitinating enzyme family, is abnormally expressed in breast cancer tissues (especially TNBC) and is closely related to the occurrence and development of breast cancer. Clinical data show that patients with high USP16 expression have shorter disease-free survival and overall survival, and an increased risk of tumor recurrence. This suggests that USP16 is not only a potential prognostic biomarker for breast cancer (especially TNBC), but also a highly promising therapeutic target.

[0004] Preliminary progress has been made in the research of small molecule inhibitors targeting USP16. For example, the compound IU1 can inhibit the deubiquitinating enzyme activity of USP16 by binding to its catalytic domain, effectively inhibiting breast cancer cell proliferation and migration and enhancing chemosensitivity in preclinical models. However, existing USP16 inhibitors (such as IU1) still have significant limitations: on the one hand, their inhibitory activity and selectivity need to be improved, and off-target effects may lead to safety risks; on the other hand, most inhibitors are still in the preclinical research stage, lacking highly targeted and druggable candidate molecules. Therefore, the development of novel USP16 inhibitors with high selectivity and low toxicity remains of significant scientific importance. Summary of the Invention

[0005] To enhance the therapeutic effect of breast cancer, this invention provides an acylhydrazine compound with antitumor activity, its preparation method, and its application.

[0006] The first aspect of the present invention provides an acylhydrazine compound or a pharmaceutically acceptable salt thereof, the acylhydrazine compound having the structure shown in Formula 3:

[0007] ;

[0008] Among them, R 1 R is a mono- or poly-substituted group on the benzene ring. 1 Selected from C1-C4 alkyl groups, C1-C4 alkoxy groups, and halogen atoms; R 2 Selected from C1 to C4 alkyl, hydrogen, and halogen atoms.

[0009] The aforementioned acylhydrazine compounds, or pharmaceutically acceptable salts thereof, possess the property of selectively inhibiting USP16 and can be used to prepare USP16 inhibitors. Pharmaceutically acceptable salts of the aforementioned acylhydrazine compounds can be readily prepared from the aforementioned acylhydrazine compounds as starting materials; for example, reacting the aforementioned acylhydrazine compounds with hydrochloric acid can yield the hydrochloride salts of the aforementioned acylhydrazine compounds.

[0010] In some alternative embodiments, the above-mentioned R 1 Selected from methyl, ethyl, methoxy, ethoxy, fluorine, chlorine, bromine, and iodine atoms; The above R 2 Selected from methyl, ethyl, hydrogen, fluorine, chlorine, bromine, and iodine atoms.

[0011] In some alternative embodiments, the above-mentioned acylhydrazine compound has a structure shown in one of formulas 3a-3k:

[0012] , , , , , , , , , , .

[0013] A second aspect of the present invention provides a method for preparing the above-mentioned acylhydrazine compound, comprising the following steps:

[0014] In an organic solvent, the compound shown in Formula 1 undergoes an amide condensation reaction with the compound shown in Formula 2 to obtain the acylhydrazine compound shown in Formula 3.

[0015] , ;

[0016] Among them, R 1 R 2 The definition is the same as above.

[0017] In some alternative embodiments, the above-mentioned amide condensation reaction is carried out in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine.

[0018] In some optional embodiments, the above-mentioned amide condensation reaction is carried out at 0~80°C.

[0019] In some optional embodiments, the organic solvent described above may be N,N-dimethylformamide.

[0020] A third aspect of the present invention provides the use of the above-mentioned acylhydrazine compound or a pharmaceutically acceptable salt thereof in the preparation of a USP16 inhibitor.

[0021] A fourth aspect of the present invention provides a USP16 inhibitor comprising the above-described acylhydrazine compound or a pharmaceutically acceptable salt thereof.

[0022] The fifth aspect of the present invention provides the use of the above-mentioned acylhydrazine compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing breast cancer.

[0023] In some alternative embodiments, the aforementioned breast cancer cell line may be MCF-7 or MDA-MB-231.

[0024] A sixth aspect of the present invention provides a medicament for treating and / or preventing breast cancer, comprising the aforementioned acylhydrazine compound or a pharmaceutically acceptable salt thereof. The aforementioned acylhydrazine compound or a pharmaceutically acceptable salt thereof is the main active ingredient (principal drug) in the aforementioned medicament for treating and / or preventing breast cancer.

[0025] In some optional embodiments, the above-mentioned medicaments for treating and / or preventing breast cancer also include excipients. The dosage form of the above-mentioned medicaments for treating and / or preventing breast cancer is any pharmaceutically acceptable dosage form. The above-mentioned excipients are stable in nature, have no incompatibility with the active pharmaceutical ingredient, do not produce side effects, do not affect efficacy, are not easily deformed, cracked, or moldy at room temperature, and are harmless to the human body.

[0026] In some alternative embodiments, the excipient is at least one selected from gum arabic, syrup, lanolin, and starch.

[0027] The technical solutions of the embodiments of the present invention have the following beneficial effects:

[0028] The acylhydrazine compound in this invention can be used as a USP16 inhibitor, exhibiting highly efficient activity in inhibiting the proliferation of breast cancer cells, thus providing a safe and efficient candidate drug molecule for the treatment of breast cancer. The synthesis method of this acylhydrazine compound is simple, the raw materials are readily available, and the functional groups have good compatibility. Experimental results show that the tumor inhibition rate of this acylhydrazine compound in the mouse tumor-bearing group is better than that in the positive control group, indicating good application prospects. Attached Figure Description

[0029] Figure 1This is a graph showing the tumor volume changes in the MDA-MB-231 cell tumor model mouse in Example 14 of this invention.

[0030] Figure 2 This is a graph showing the tumor volume changes in the MCF-7 cell colony model mice in Example 14 of this invention.

[0031] Figure 3 This is a graph showing the weight changes of mice with the MDA-MB-231 cell tumor model in Example 14 of this invention.

[0032] Figure 4 This is a graph showing the weight changes of the MCF-7 cell tumor model mice in Example 14 of this invention.

[0033] Figure 5 This is a graph showing the changes in tumor weight in the MDA-MB-231 cell tumor model mice in Example 14 of this invention.

[0034] Figure 6 This is a diagram showing the tumor changes in the MCF-7 cell colony model mouse in Example 14 of this invention. Detailed Implementation

[0035] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0036] In the following examples, the synthetic route of the acylhydrazine compound is as follows:

[0037]

[0038] Among them, R 1 R is a mono- or poly-substituted group on the benzene ring. 1 Selected from C1-C4 alkyl groups, C1-C4 alkoxy groups, and halogen atoms; R 2 Selected from C1 to C4 alkyl, hydrogen, and halogen atoms.

[0039] The synthetic route for the above-mentioned acylhydrazine compounds specifically includes the following steps:

[0040] The substituted phenethylhydrazine compound shown in Formula 1 (1.0 mmol, 1.0 eq) and the substituted 2-thiophenecarboxylic acid shown in Formula 2 (1.0 mmol, 1.0 eq) were dissolved in 10 mL of N,N-dimethylformamide. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 mmol, 1.0 eq), 1-hydroxybenzotriazole (1.0 mmol, 1.0 eq), and N,N-diisopropylethylamine (2.0 mmol, 2.0 eq) were added sequentially to the system, and the reaction was carried out at room temperature for 6 hours. The reaction was quenched with 100 mL of water, and the mixture was extracted three times with ethyl acetate (50.0 mL). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was evaporated under vacuum to remove the solvent, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate 1:1 ~ dichloromethane: methanol = 20:1) to obtain the target product.

[0041] Only the substituent R in the phenylethylhydrazine-like compounds shown in Formula 1 is changed. 1 The substituent R in the substituted 2-thiophenecarboxylic acid shown in Formula 2 2 A series of acylhydrazine compounds as shown in Formula 3 can be obtained.

[0042] Example 1: Synthesis of the acylhydrazine compound shown in formula 3a

[0043]

[0044] This example yielded 226.8 mg of the acylhydrazine compound of formula 3a (yield 81%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d ) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J = 5.3, 1.6 Hz, 1H), 7.33 – 7.29 (m, 2H), 7.19 (dt, J = 8.2, 1.1 Hz, 2H), 7.14 (dd, J = 6.5, 5.4 Hz, 1H), 5.78 (dt, J = 4.9, 3.6Hz, 1H), 3.08 (td, J = 5.5, 3.7 Hz, 2H), 2.81 (tt, J = 5.6, 1.1 Hz, 2H). 13CNMR (100 MHz, Chloroform- d ) δ 162.1, 136.9, 136.7, 132.7, 132.0, 130.4,129.1, 128.5, 128.2, 49.5, 34.0.

[0045] Example 2: Synthesis of the acylhydrazine compound shown in formula 3b

[0046]

[0047] This example yielded 209.8 mg of the acylhydrazine compound shown in formula 3b (yield 76%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d ) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J = 5.3, 1.6 Hz, 1H), 7.14 (dd, J = 6.5, 5.4 Hz, 1H), 7.05 (dt, J = 8.5, 1.0 Hz, 2H), 6.88 – 6.82 (m, 2H), 5.78 (dt, J = 4.9, 3.7Hz, 1H), 3.78 (s, 3H), 3.08 (td, J = 5.5, 3.7 Hz, 2H), 2.81 (tt, J = 5.5, 0.9Hz, 2H). 13 C NMR (100 MHz, Chloroform- d ) δ 162.1, 158.3, 136.7, 132.1, 132.0,129.6, 129.1, 128.2, 113.6, 55.3, 49.5, 34.0.

[0048] Example 3: Synthesis of the acylhydrazine compound shown in formula 3c

[0049]

[0050] This example yielded 223.4 mg of the acylhydrazine compound shown in formula 3c (yield 73%), and its characterization results are as follows: 1HNMR (400 MHz, Chloroform- d ) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J = 5.3, 1.6 Hz, 1H), 7.14 (dd, J = 6.5, 5.4 Hz, 1H), 6.76 (d, J = 8.4 Hz, 1H), 6.70 (dt, J = 1.9, 1.0 Hz, 1H), 6.64 (ddt, J = 8.4,2.0, 1.0 Hz, 1H), 5.77 (dt, J = 4.8, 3.7 Hz, 1H), 3.82 (d, J = 4.6 Hz, 6H), 3.10 (td, J = 5.5, 3.7 Hz, 2H), 2.79 (tt, J = 5.3, 1.0 Hz, 2H). 13 C NMR (100MHz, Chloroform- d ) δ 162.1, 149.1, 147.9, 136.7, 132.4, 132.0, 129.1, 128.2,122.2, 112.6, 111.9, 55.9, 55.8, 49.5, 34.0.

[0051] Example 4: Synthesis of the acylhydrazine compound shown in formula 3d

[0052]

[0053] This example yielded 249.5 mg of the acylhydrazine compound of formula 3d (yield 77%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d ) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J= 5.3, 1.6 Hz, 1H), 7.40 – 7.34 (m, 2H), 7.20 (dt, J = 8.2, 1.0 Hz, 2H), 7.14 (dd, J = 6.5, 5.4 Hz, 1H), 5.78 (dt, J = 4.9, 3.7Hz, 1H), 3.08 (td, J = 5.5, 3.7 Hz, 2H), 2.80 (tt, J = 5.5, 1.0 Hz, 2H). 13 CNMR (100 MHz, Chloroform- d ) δ 162.1, 137.6, 136.7, 132.1, 131.3, 130.6,129.1, 128.2, 120.0, 49.5, 33.9.

[0054] Example 5: Synthesis of the acylhydrazine compound shown in formula 3e

[0055]

[0056] This example yielded 198.0 mg of the acylhydrazine compound shown in formula 3e (yield 75%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d ) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J = 5.3, 1.6 Hz, 1H), 7.36 – 7.28 (m, 1H), 7.24 – 7.07(m, 4H), 5.54 (dt, J = 4.9, 3.6 Hz, 1H), 3.07 (td, J = 5.3, 3.7 Hz, 2H), 2.90(td, J = 5.3, 0.9 Hz, 2H). 13 C NMR (100 MHz, Chloroform- d) δ 162.5, 162.1,160.5, 136.7, 132.0, 131.0, 131.0, 129.1, 128.2, 128.2, 126.8, 126.6, 124.8,124.8, 115.2, 115.1, 48.2, 48.2, 29.1, 29.1.

[0057] Example 6: Synthesis of the acylhydrazine compound shown in formula 3f

[0058]

[0059] This example yielded 187.2 mg of the acylhydrazine compound of formula 3f (yield 72%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d ) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J = 5.3, 1.6 Hz, 1H), 7.17 – 7.08 (m, 4H), 7.08 – 7.03(m, 1H), 5.58 (dt, J = 4.8, 3.7 Hz, 1H), 3.05 (td, J = 5.3, 3.6 Hz, 2H), 2.87(td, J = 5.3, 1.0 Hz, 2H), 2.31 (s, 3H). 13 C NMR (100 MHz, Chloroform- d ) δ162.1, 138.5, 137.2, 136.7, 132.0, 130.1, 129.2, 129.1, 128.2, 126.7, 48.4,31.0, 20.7.

[0060] Example 7: Synthesis of the acylhydrazine compound shown in formula 3g

[0061]

[0062] This example yielded 192.7 mg of the acylhydrazine compound shown in formula 3 g (yield 73%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J = 5.3, 1.6 Hz, 1H), 7.23 (ddt, J = 8.2, 5.1, 1.0 Hz,2H), 7.14 (dd, J = 6.5, 5.4 Hz, 1H), 7.08 – 7.00 (m, 2H), 5.78 (dt, J = 4.9, 3.6 Hz, 1H), 3.08 (td, J = 5.5, 3.7 Hz, 2H), 2.81 (tt, J = 5.5, 0.9 Hz, 2H). 13 C NMR (100 MHz, Chloroform- d ) δ 162.5, 162.1, 160.5, 136.7, 134.6, 134.5,132.0, 130.5, 130.4, 129.1, 128.2, 115.2, 115.1, 49.5, 33.3.

[0063] Example 8: Synthesis of the acylhydrazine compound shown in formula 3h

[0064]

[0065] This example yielded 184.6 mg of the acylhydrazine compound shown in Formula 3h (yield 71%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d ) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J = 5.3, 1.7 Hz, 1H), 7.17 – 7.11 (m, 2H), 7.11 – 7.08(m, 2H), 7.04 (dt, J = 8.0, 1.0 Hz, 2H), 5.77 (dt, J = 4.9, 3.7 Hz, 1H), 3.07(td,J = 5.5, 3.7 Hz, 2H), 2.80 (tt, J = 5.5, 1.0 Hz, 2H), 2.34 (s, 3H). 13 CNMR (100 MHz, Chloroform- d ) δ 162.1, 137.0, 136.7, 136.3, 132.0, 129.1,129.0, 128.8, 128.2, 49.5, 33.7, 21.0.

[0066] Example 9: Synthesis of the acylhydrazine compound shown in formula 3i

[0067]

[0068] This example yielded 215.6 mg of the acylhydrazine compound of formula 3i (yield 77%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d ) δ 8.49 (d, J = 4.9 Hz, 1H), 7.86 (dd, J = 6.5, 1.7 Hz, 1H), 7.74 (dd, J = 5.3, 1.6 Hz, 1H), 7.36 – 7.30 (m, 1H), 7.27 – 7.21(m, 3H), 7.21 – 7.11 (m, 2H), 5.53 (dt, J = 4.8, 3.6 Hz, 1H), 3.09 – 3.02 (m, 2H), 2.96 – 2.90 (m, 2H). 13 C NMR (100 MHz, Chloroform- d ) δ 162.1, 137.1,136.7, 134.5, 132.0, 130.7, 129.3, 129.1, 128.5, 128.2, 127.8, 48.4, 30.9.

[0069] Example 10: Synthesis of the acylhydrazine compound shown in formula 3j

[0070]

[0071] This example yielded 211.7 mg of the acylhydrazine compound shown in formula 3j (yield 72%), and its characterization results are as follows:1 HNMR (400 MHz, Chloroform- d ) δ 8.27 (d, J = 4.9 Hz, 1H), 7.65 (d, J = 6.6 Hz,1H), 7.36 – 7.30 (m, 1H), 7.27 – 7.17 (m, 3H), 6.78 (dq, J = 6.7, 0.8 Hz, 1H), 5.53 (dt, J = 4.8, 3.6 Hz, 1H), 3.09 – 3.02 (m, 2H), 2.96 – 2.90 (m,2H), 2.52 (s, 3H). 13 C NMR (100 MHz, Chloroform- d ) δ 161.9, 145.7, 137.2,137.1, 134.5, 130.7, 130.6, 129.3, 128.5, 127.8, 127.7, 48.4, 30.9, 16.0.

[0072] Example 11: Synthesis of the acylhydrazine compound shown in formula 3k

[0073]

[0074] This example yielded 232.4 mg of the acylhydrazine compound of formula 3k (yield 74%), and its characterization results are as follows: 1 HNMR (400 MHz, Chloroform- d ) δ 8.50 (d, J = 4.9 Hz, 1H), 7.57 (d, J = 6.8 Hz,1H), 7.36 – 7.30 (m, 1H), 7.27 – 7.17 (m, 3H), 7.03 (d, J = 6.8 Hz, 1H), 5.55(dt, J = 5.0, 3.7 Hz, 1H), 3.09 – 3.02 (m, 2H), 2.96 – 2.90 (m, 2H). 13 C NMR (100 MHz, Chloroform- d) δ 161.4, 137.4, 137.1, 134.5, 134.1, 130.7, 129.3,128.5, 128.5, 127.8, 127.3, 48.4, 30.9.

[0075] Example 12: Test of USPs kinase inhibition rate of acylhydrazine compound

[0076] The acylhydrazide compounds shown in formulas 3a-3k were first diluted to 10 μM with deionized water. Then, the optimal concentration (2 nM) of deubiquitinating enzyme (DUB) was added, and after brief mixing, the mixture was placed in a 200 μL reaction volume and incubated at room temperature for 1 hour. The system contained 50 mM HEPES (pH 8.0), 150 mM NaCl, 0.5 mM EDTA, 1 mM DTT, and 0.1 mg / mL bovine serum albumin (BSA). Next, 200 nM ubiquitin-rhodamine 110 (Ub-Rho110, catalog number #M3022, UPBio) was added, and the final fluorescence signal (excitation wavelength: 485 nm, emission wavelength: 535 nm) was acquired using a TECAN SPARK multi-mode microplate reader. The inhibition rate of the compound on enzyme activity was then calculated.

[0077] The test results are shown in Table 1. All the acylhydrazine compounds shown in Formulas 3a-3k can selectively inhibit the kinase activity of USP16 at the protein level.

[0078] Table 1. Results of USPs kinase inhibition rate test of acylhydrazine compounds (%)

[0079]

[0080] Example 13: Determination of the in vitro antiproliferative activity of acylhydrazine compounds against breast cancer cell lines

[0081] The in vitro antiproliferative activity of the acylhydrazine compounds shown in formulas 3b, 3c, 3d, 3f, 3g, 3h, 3j, and 3k against breast cancer cell lines (MCF-7 and MDA-MB-231) was evaluated using a CCK-8 assay. Ten concentration gradients of the acylhydrazine compounds were established, with a maximum concentration of 10 µM, followed by 3-fold serial dilutions, and a minimum concentration of 0 μM. These solutions were co-incubated with MCF-7 and MDA-MB-231 cells for 72 h. Cell viability was assessed using CCK-8, and data were processed using GraphPad Prism 9 to calculate the IC50. 50 value.

[0082] The results are shown in Table 2. The acylhydrazine compounds represented by formulas 3b, 3c, 3d, 3f, 3g, 3h, 3j, and 3k all exhibited excellent in vitro anti-breast cancer cell proliferation activity. Among them, the acylhydrazine compounds represented by formulas 3b, 3f, and 3j showed the best activity.

[0083] Table 2. Results of the determination of the inhibitory activity of acylhydrazine compounds on the proliferation of MCF-7 and MDA-MB-231.

[0084]

[0085] Example 14: Animal Tumor Suppression Experiment

[0086] First, MCF-7 and MDA-MB-231 cells were cultured in a 37°C, 5% CO2 incubator. BALB / c mice were housed under standard specific pathogen-free (SPF) conditions. A xenograft tumor model was established by subcutaneously injecting MCF-7 and MDA-MB-231 cells into 6-8 week old BALB / c mice. Tumor formation was examined daily to determine tumor development. Once the tumors reached a certain size, mice were randomly divided into a control group and a treatment group (n=5 per group). Mice in the treatment group received an intraperitoneal injection of an aqueous solution of the acylhydrazine compounds shown in formulas 3b, 3f, and 3j at a dose of 50 mpk. Tumor diameter was measured daily using calipers, and mouse weight was recorded. Tumor tissue was harvested on day 21, and the tumor volume change was used to evaluate the antitumor effect of the compound.

[0087] The experimental results are shown in Table 3 and Figures 1-6 As shown, the acylhydrazine compounds represented by formulas 3b, 3f, and 3j all exhibit excellent in vivo antiproliferative activity.

[0088] Table 3. Results of animal tumor inhibition rate experiment (%)

[0089]

[0090] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any form or substance. It should be noted that those skilled in the art can make various improvements and additions without departing from the method of the present invention, and these improvements and additions should also be considered within the scope of protection of the present invention. Any modifications, alterations, and equivalent changes made by those skilled in the art based on the above-disclosed technical content without departing from the spirit and scope of the present invention are equivalent embodiments of the present invention. Furthermore, any modifications, alterations, and evolutions made to the above embodiments based on the essential technology of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. An acylhydrazine compound or a pharmaceutically acceptable salt thereof, characterized in that, The acylhydrazine compound has the structure shown in Formula 3: ； Among them, R 1 R is a mono- or poly-substituted group on the benzene ring. 1 Selected from C1-C4 alkyl groups, C1-C4 alkoxy groups, and halogen atoms; R 2 Selected from C1 to C4 alkyl, hydrogen, and halogen atoms.

2. The acylhydrazine compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The R 1 Selected from methyl, ethyl, methoxy, ethoxy, fluorine, chlorine, bromine, and iodine atoms; The R 2 Selected from methyl, ethyl, hydrogen, fluorine, chlorine, bromine, and iodine atoms.

3. The acylhydrazine compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The acylhydrazine compound has a structure shown in one of formulas 3a-3k: 、 、 、 、 、 、 、 、 、 、 。 4. The method for preparing the acylhydrazine compound according to claim 1, characterized in that, Includes the following steps: In an organic solvent, the compound shown in Formula 1 undergoes an amide condensation reaction with the compound shown in Formula 2 to obtain the acylhydrazine compound shown in Formula 3. ， ； Among them, R 1 R is a mono- or poly-substituted group on the benzene ring. 1 Selected from C1-C4 alkyl groups, C1-C4 alkoxy groups, and halogen atoms; R 2 Selected from C1 to C4 alkyl, hydrogen, and halogen atoms.

5. The method according to claim 4, characterized in that, The amide condensation reaction was carried out in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine.

6. The method according to claim 4, characterized in that, The amide condensation reaction is carried out at 0~80℃.

7. The use of any of the acylhydrazine compounds according to claims 1-3 or a pharmaceutically acceptable salt thereof in the preparation of USP16 inhibitors.

8. A USP16 inhibitor, characterized in that, Includes any of the acylhydrazine compounds described in claims 1-3 or their pharmaceutically acceptable salts.

9. The use of any of the acylhydrazine compounds according to claims 1-3 or a pharmaceutically acceptable salt thereof in the preparation of medicaments for the treatment and / or prevention of breast cancer.

10. A medicament for treating and / or preventing breast cancer, characterized in that, Includes any of the acylhydrazine compounds described in claims 1-3 or their pharmaceutically acceptable salts.

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