Acylhydrazine compound with anti-tumor activity as well as preparation method and application of acylhydrazine compound

Abstract

The invention relates to the technical field of biological medicine, in particular to an acylhydrazine compound with anti-tumor activity and a preparation method and application thereof. In the invention, the acylhydrazine compound is prepared, and the acylhydrazine compound can be used as a USP16 inhibitor, has the activity of efficiently inhibiting breast cancer cell proliferation, and provides a safe and efficient candidate drug molecule for treating breast cancer. The synthesis method of the acylhydrazine compound is simple, raw materials are easy to obtain, and functional groups are good in compatibility. Experimental results show that the tumor inhibition rate of the acylhydrazine compound for treating a mouse tumor-bearing group is superior to that of a positive control group, and the acylhydrazine compound has a good application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to an acylhydrazine compound with anti-tumor activity, a preparation method and application thereof. Background Art

[0002] Breast cancer is a serious malignancy threatening women's health worldwide. It is highly heterogeneous and can be divided into molecular subtypes, including luminal A, luminal B, HER2-overexpressing, and triple-negative breast cancer (TNBC), based on the expression of estrogen receptors (ER), progesterone receptors (PR), and human epidermal growth factor receptor 2 (HER2). TNBC, lacking ER, PR, and HER2 targets, is insensitive to endocrine therapy and HER2-targeted therapies, resulting in high recurrence rates and poor prognosis, making it a challenge in clinical treatment. Therefore, in-depth understanding of the molecular mechanisms underlying breast cancer development and progression, as well as 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, particularly TNBC, and is closely associated with the development and progression of breast cancer. Clinical data show that patients with high USP16 expression have shortened disease-free and overall survival, and an increased risk of tumor recurrence. This suggests that USP16 is not only a potential prognostic marker for breast cancer, particularly TNBC, but also a promising therapeutic target.

[0004] Initial progress has been made in the study of small molecule inhibitors targeting USP16. For example, compound IU1 inhibits the deubiquitinase activity of USP16 by binding to its catalytic domain, effectively inhibiting breast cancer cell proliferation and migration and enhancing chemotherapy sensitivity in preclinical models. However, existing USP16 inhibitors, such as IU1, still have significant limitations. Firstly, their inhibitory activity and selectivity need to be improved, and off-target effects may pose safety risks. Secondly, the vast majority of inhibitors are still in the preclinical development stage, lacking highly targeted and druggable candidate molecules. Therefore, the development of novel USP16 inhibitors with high selectivity and low toxicity remains of great scientific significance. Summary of the Invention

[0005] In order to promote the therapeutic effect of breast cancer, the present invention provides an acylhydrazine compound with anti-tumor activity, a preparation method and application thereof.

[0006] The first aspect of the present invention provides an acylhydrazine compound or a pharmaceutically acceptable salt thereof, wherein the acylhydrazine compound has a structure shown in Formula 3: ; Among them, R 1is a single or multiple substituted group on the benzene ring, R 1 is selected from C1~C4 alkyl, C1~C4 alkoxy, and halogen atom; R 2 selected from C1~C4 alkyl groups, hydrogen atoms, and halogen atoms.

[0007] The aforementioned acylhydrazine compounds or pharmaceutically acceptable salts thereof have the property of selectively inhibiting USP16 and can be used to prepare USP16 inhibitors. The pharmaceutically acceptable salts of the aforementioned acylhydrazine compounds can be easily prepared using the aforementioned acylhydrazine compounds as raw materials. For example, the hydrochloride salt of the aforementioned acylhydrazine compound can be prepared by reacting the aforementioned acylhydrazine compound with hydrochloric acid.

[0008] In some optional embodiments, the above R 1 is selected from the group consisting of methyl, ethyl, methoxy, ethoxy, fluorine, chlorine, bromine, and iodine atoms; 2 It is selected from a methyl group, an ethyl group, a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

[0009] In some optional embodiments, the acylhydrazine compound has a structure shown in one of Formula 3a to Formula 3k: 、 、 、 、 、 、 、 、 、 、 .

[0010] A second aspect of the present invention provides a method for preparing the above-mentioned acylhydrazine compound, which comprises the following steps: In an organic solvent, the compound represented by Formula 1 and the compound represented by Formula 2 undergo an amide condensation reaction to obtain an acylhydrazine compound represented by Formula 3; , ; Among them, R 1 、R 2 The definition of is the same as above.

[0011] In some optional embodiments, the amide condensation reaction is carried out in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine.

[0012] In some optional embodiments, the amide condensation reaction is carried out at 0-80°C.

[0013] In some optional embodiments, the organic solvent may be N,N-dimethylformamide.

[0014] The third aspect of the present invention provides use of the above-mentioned acylhydrazine compound or a pharmaceutically acceptable salt thereof in the preparation of a USP16 inhibitor.

[0015] The fourth aspect of the present invention provides a USP16 inhibitor comprising the above-mentioned acylhydrazine compound or a pharmaceutically acceptable salt thereof.

[0016] A fifth aspect of the present invention provides 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.

[0017] In some optional embodiments, the breast cancer cell line may be MCF-7 or MDA-MB-231.

[0018] A sixth aspect of the present invention provides a drug 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 (primary agent) in the drug for treating and / or preventing breast cancer.

[0019] In some optional embodiments, the drug for treating and / or preventing breast cancer further comprises an excipient. The drug for treating and / or preventing breast cancer can be in any pharmaceutically acceptable dosage form. The excipient is stable, has no incompatibility with the main drug, does not produce side effects, does not affect efficacy, is not easily deformed, cracked, or moldy at room temperature, and is harmless to the human body.

[0020] In some optional embodiments, the excipient is at least one of gum arabic, syrup, lanolin, and starch.

[0021] The technical solution of the embodiment of the present invention has the following beneficial effects: The acylhydrazine compound in the embodiment of the present invention can be used as a USP16 inhibitor, has the activity of effectively inhibiting the proliferation of breast cancer cells, and provides a safe and efficient candidate drug molecule for the treatment of breast cancer; the synthesis method of the acylhydrazine compound is simple, the raw materials are readily available, and the functional group compatibility is good; experimental results show that the tumor inhibition rate of the tumor-bearing group of mice treated with the acylhydrazine compound is better than that of the positive control group, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a graph showing the changes in tumor volume in mice bearing the MDA-MB-231 cell tumor model in Example 14 of the present invention.

[0023] Figure 2This is a graph showing the changes in tumor volume in mice bearing the MCF-7 cell-derived tumor model in Example 14 of the present invention.

[0024] Figure 3 This is a graph showing weight changes in mice bearing the MDA-MB-231 cell tumor model in Example 14 of the present invention.

[0025] Figure 4 This is a graph showing weight changes in mice bearing the MCF-7 cell tumor model in Example 14 of the present invention.

[0026] Figure 5 This is a graph showing changes in tumor weight in mice harboring the MDA-MB-231 cell-derived tumor model in Example 14 of the present invention.

[0027] Figure 6 This is a diagram showing the tumor changes in the MCF-7 cell-derived tumor model mice in Example 14 of the present invention. DETAILED DESCRIPTION

[0028] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0029] In the following examples, the synthetic routes of acylhydrazine compounds are as follows:

[0030] Among them, R 1 is a single or multiple substituted group on the benzene ring, R 1 is selected from C1~C4 alkyl, C1~C4 alkoxy, and halogen atom; R 2 selected from C1~C4 alkyl groups, hydrogen atoms, and halogen atoms.

[0031] The synthetic route of the above-mentioned acylhydrazine compound specifically comprises the following steps: The substituted phenelzizine compound (1.0 mmol, 1.0 eq) of Formula 1 and the substituted 2-thiophenecarboxylic acid (1.0 mmol, 1.0 eq) of Formula 2 were dissolved in 10 mL of N,N-dimethylformamide. 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 then added sequentially to the system and reacted at room temperature for 6 hours. The reaction was quenched by the addition of 100 mL of water and extracted three times with ethyl acetate (50.0 mL). The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by vortexing the organic phases under vacuum. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1 to dichloromethane:methanol = 20:1) to obtain the desired product.

[0032] Only the substituent R in the substituted phenelzine compound shown in formula 1 is changed 1 and the substituent R in the substituted 2-thiophenecarboxylic acid shown in formula 2 2 , a series of acylhydrazine compounds shown in formula 3 can be obtained.

[0033] Example 1: Synthesis of the acylhydrazine compound represented by formula 3a

[0034] In this example, 226.8 mg of the acylhydrazine compound represented by formula 3a was obtained (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. Example 2: Synthesis of the acylhydrazine compound represented by formula 3b

[0035] In this example, 209.8 mg of the acylhydrazine compound represented by Formula 3b was obtained (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, 0.9 Hz, 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. Example 3: Synthesis of the acylhydrazine compound represented by formula 3c

[0036] In this example, 223.4 mg of the acylhydrazine compound represented by formula 3c was obtained (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), 7.14 (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. Example 4: Synthesis of the acylhydrazine compound represented by formula 3d

[0037] In this example, 249.5 mg of the acylhydrazine compound represented by Formula 3d was obtained (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. Example 5: Synthesis of the acylhydrazine compound represented by formula 3e

[0038] In this example, 198.0 mg of the acylhydrazine compound represented by Formula 3e was obtained (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. Example 6: Synthesis of the acylhydrazine compound represented by formula 3f

[0039] In this example, 187.2 mg of the acylhydrazine compound represented by Formula 3f was obtained (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. Example 7: Synthesis of the acylhydrazine compound shown in formula 3g

[0040] In this example, 192.7 mg of the acylhydrazine compound represented by Formula 3g was obtained (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. Example 8: Synthesis of the acylhydrazine compound represented by formula 3h

[0041] In this example, 184.6 mg of the acylhydrazine compound represented by formula 3h was obtained (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. Example 9: Synthesis of the acylhydrazine compound represented by formula 3i

[0042] In this example, 215.6 mg of the acylhydrazine compound represented by formula 3i was obtained (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. Example 10: Synthesis of the acylhydrazine compound represented by formula 3j

[0043] In this example, 211.7 mg of the acylhydrazine compound represented by Formula 3j was obtained (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). 13C 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. Example 11: Synthesis of the acylhydrazine compound represented by formula 3k

[0044] In this example, 232.4 mg of the acylhydrazine compound represented by formula 3k was obtained (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. Example 12: Acylhydrazine Compound USPs Kinase Inhibition Rate Test Acylhydrazide compounds represented by Formulas 3a-3k were diluted to 10 μM in deionized water. Deubiquitinase (DUB) was then added at an optimal concentration (2 nM). After brief mixing, the mixture was placed in a 200 μL reaction system containing 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). The reaction was incubated at room temperature for 1 hour. Next, 200 nM ubiquitin rhodamine 110 (Ub-Rho110, Catalog # M3022, UBP Bio) was added, and the resulting fluorescence signal was acquired using a TECAN SPARK multi-function microplate reader (excitation wavelength: 485 nm, emission wavelength: 535 nm). The inhibition rate of the enzyme activity was calculated based on the fluorescence signal.

[0045] The test results are shown in Table 1. The acylhydrazine compounds represented by Formula 3a to Formula 3k can selectively inhibit the kinase activity of USP16 at the protein level.

[0046] Table 1 Kinase inhibition test results of acylhydrazine compounds USPs (%)

[0047] Example 13: In vitro antiproliferative activity assay of acylhydrazide compounds against breast cancer cell lines The in vitro antiproliferative activity of the acylhydrazine compounds represented by 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. The acylhydrazine compound solution was diluted three-fold to a minimum concentration of 0 μM, with a total of 10 concentration gradients. MCF-7 and MDA-MB-231 cells were incubated for 72 h. Cell viability was assessed using CCK-8 assays, and data were processed using GraphPad Prism 9 to calculate the IC values. 50 value.

[0048] The test results are shown in Table 2. The acylhydrazine compounds represented by Formulas 3b, 3c, 3d, 3f, 3g, 3h, 3j, and 3k all have excellent in vitro anti-breast cancer cell proliferation activity, among which the acylhydrazine compounds represented by Formulas 3b, 3f, and 3j have the best activity.

[0049] Table 2 Results of the assay of the inhibitory activity of acylhydrazide compounds on the proliferation of MCF-7 and MDA-MB-231

[0050] Example 14: Animal tumor inhibition experiment MCF-7 and MDA-MB-231 cells were cultured at 37°C in a 5% CO2 incubator. BALB / c mice were housed under standard specific pathogen-free (SPF) conditions. Six- to eight-week-old BALB / c mice were subcutaneously injected with MCF-7 and MDA-MB-231 cells to establish xenograft tumor models. Mice were examined daily to assess tumor progression. When tumors reached a certain size, mice were randomly divided into a control group and a treatment group, with five mice in each group. The treatment groups received an intraperitoneal injection of 50 mpk of an aqueous solution of the acylhydrazine compounds represented by Formulas 3b, 3f, and 3j. Tumor diameters were measured daily with a vernier caliper and mouse weights were recorded. Tumor tissue was harvested on day 21, and the anti-tumor effect of the compound was evaluated by changes in tumor volume.

[0051] 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 have excellent in vivo anti-proliferative activity.

[0052] Table 3 Animal tumor inhibition rate experimental results (%)

[0053] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form or substance. It should be pointed out that ordinary technicians in this technical field can make several improvements and supplements without departing from the method of the present invention. These improvements and supplements should also be regarded as the scope of protection of the present invention. Any equivalent changes, modifications and evolutions made by technicians familiar with this profession without departing from the spirit and scope of the present invention by using the technical content disclosed above are all equivalent embodiments of the present invention; at the same time, any equivalent changes, modifications and evolutions made to the above embodiments based on the essential technology of the present invention are still 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 a structure shown in Formula 3: ； Among them, R 1 is a single or multiple substituted group on the benzene ring, R 1 is selected from C1~C4 alkyl, C1~C4 alkoxy, and halogen atom; R 2 selected from C1~C4 alkyl groups, hydrogen atoms, and halogen atoms.

2. The acylhydrazine compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein The R 1 is selected from the group consisting of methyl, ethyl, methoxy, ethoxy, fluorine, chlorine, bromine, and iodine atoms; 2 It is selected from a methyl group, an ethyl group, a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

3. The acylhydrazine compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The acylhydrazine compound has a structure shown in one of Formula 3a to Formula 3k: 、 、 、 、 、 、 、 、 、 、 。 4. The method for preparing an acylhydrazine compound according to claim 1, wherein The following steps are involved: In an organic solvent, the compound represented by Formula 1 and the compound represented by Formula 2 undergo an amide condensation reaction to obtain an acylhydrazine compound represented by Formula 3; ， ； Among them, R 1 is a single or multiple substituted group on the benzene ring, R 1 is selected from C1~C4 alkyl, C1~C4 alkoxy, and halogen atom; R 2 selected from C1~C4 alkyl groups, hydrogen atoms, and halogen atoms.

5. The method according to claim 4, characterized in that The amide condensation reaction is 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 a temperature of 0-80°C.

7. Use of the acylhydrazine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a USP16 inhibitor.

8. A USP16 inhibitor, characterized in that The invention comprises the acylhydrazine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3.

9. Use of the acylhydrazine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a medicament for treating and / or preventing breast cancer.

10. A drug for treating and / or preventing breast cancer, characterized in that: The invention comprises the acylhydrazine compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3.

Images