Derivative small molecules containing indole group and triazine group and application thereof

Abstract

The invention belongs to the technical field of medicinal chemistry, and discloses a derivative small molecule containing an indole group and a triazine group and application of the derivative small molecule in preparation of an NAE / HDAC double-target inhibitor. The derivative small molecule containing the indole group and the triazine group disclosed by the invention is a derivative small molecule developed based on a rational pharmacophore fusion design strategy, and as a single-molecule multi-target inhibitor, the derivative small molecule can overcome the inherent technical defects of drug combination, retains the synergistic inhibition activity, and can be used for preparing a drug for preventing and treating diseases. The inhibition efficiency of part of derivative small molecules is superior to that of the existing NAE inhibitor and HDAC inhibitor combined medication.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically relating to a class of small molecules containing indole and triazine groups and their application in the preparation of NAE / HDAC dual-target inhibitors. Background Technology

[0002] Histone deacetylases (HDACs) play a crucial role in inducing tumor cell apoptosis and differentiation by removing acetylation groups from various protein substrates, altering DNA-protein interactions, and regulating the expression of tumor suppressor genes and DNA repair genes. Furthermore, research has revealed that HDACs also play a key role in numerous non-tumor pathological processes: in the cardiovascular system, specific HDAC isoenzymes are involved in the development of myocardial hypertrophy, heart failure, and atherosclerosis; in the nervous system, HDAC activity is closely related to neural plasticity and learning and memory, and its dysregulation is involved in the pathological mechanisms of neurodegenerative diseases such as Alzheimer's disease and Huntington's disease; in immune and inflammatory responses, HDACs influence the progression of autoimmune diseases such as rheumatoid arthritis and inflammatory bowel disease by regulating the expression of inflammatory factors. Since HDACs are typically overexpressed in tumor cells, HDAC inhibitors (HDACi) have emerged as a novel targeted cancer therapy modality. Currently, four drugs have been approved by the US FDA for the treatment of hematologic malignancies, including Voronostat (SAHA), Romidepsin (FK228), Belinostat (PXD10), and Panobinostat (LBH589). HDACi have been shown to have monotherapy activity in acute myeloid leukemia (AML), but in a phase II clinical trial, their monotherapy efficacy was poor, mainly because they promote the production of NF-κB (Nuclear factor kappa-B), which is beneficial to tumor cell survival. Studies have demonstrated that using proteasome-associated inhibitors can increase the lethality of HDACi against tumor cells by blocking NF-κB activation.

[0003] Neddylation is a crucial post-translational modification of proteins, catalyzed by the NEDD8-activating enzyme (NAE), which covalently links the ubiquitin molecule NEDD8 to a substrate protein. One of its core physiological functions is the activation of the Cullin-RING E3 ubiquitin ligase (CRL). The activated CRL then labels downstream substrate proteins, leading to their ubiquitination and degradation via the proteasome pathway. This precisely regulates a range of vital processes, including the cell cycle, apoptosis, DNA damage responses, and the NF-κB signaling pathway. Abnormal activity in this pathway is closely associated with various human diseases. The NAE-CRL pathway plays a critical role in a variety of physiological and pathological processes. In the cardiovascular system, it participates in regulating vascular endothelial function, smooth muscle cell behavior, and the stability of atherosclerotic plaques. In the nervous system, this pathway is crucial for maintaining protein homeostasis; its dysfunction hinders the clearance of abnormal protein aggregates, which is closely related to the development of various neurodegenerative diseases. In chronic inflammatory and fibrotic diseases (such as pulmonary fibrosis and liver fibrosis), the NAE-CRL pathway also plays an important regulatory role by regulating the expression of inflammatory mediators and pro-fibrotic factors. As the only activating enzyme in mimicry, NAE occupies a core regulatory position in this pathway and has become a promising new target in the field of anticancer drug development. The first-in-class covalent inhibitor Pevonedistat (MLN4924) can effectively inhibit NAE, leading to effects such as NF-κB inhibition, ROS accumulation, DNA replication, and damage. The completed phase I clinical trial showed that it exhibited good therapeutic potential in acute myeloid leukemia, lymphoma, melanoma, and various solid tumors, preliminarily validating the clinical safety and efficacy of NAE inhibition. The drug has now entered phase II clinical trials.

[0004] It is worth noting that NAE inhibitors and HDAC inhibitors have synergistic potential in terms of mechanism. For example, the study by Zhou et al. demonstrated that the combination of Pevonedistat and the pan-HDAC inhibitor Belinostat could produce a synergistic lethal effect on AML cells from various genetic backgrounds in vitro and improve survival rates in mouse xenograft models. This provides important evidence for developing dual-target therapeutic strategies that simultaneously target NAE and HDAC. However, the covalent action mechanism of Pevonedistat may also bring potential mechanistic toxicity. In order to expand the chemical space of NAE inhibitors and potentially obtain better pharmacokinetic properties and safety, non-covalent NAE inhibitors have become an important research direction in this field. Through reversible intermolecular forces such as hydrogen bonds, hydrophobic interactions, and van der Waals forces, NAEs such as V11 bind to NAEs. Their molecular design usually relies on a planar heterocyclic core, such as purine or triazolopyridine, that can accurately mimic adenine and occupy its binding pocket, while being supplemented with specific hydrophobic side chains to fill the hydrophobic cavity on the protein surface, thereby achieving high affinity and high selectivity inhibitory effects. These non-covalent inhibitor molecules provide a more flexible and optimized structural basis for the subsequent design and rational splicing of HDAC inhibitory pharmacophores to construct dual-target drugs. This invention abandons the traditional covalent inhibition strategy and instead focuses on non-covalent inhibitors that can reversibly bind to the NAE active site. These inhibitors typically possess a planar heterocyclic aromatic core, such as purine or triazolidine, to mimic adenine nucleosides and bind to the adenine-binding pocket of the NAE via hydrogen bonds. Simultaneously, a hydrophobic aromatic ring or alkyl chain is introduced at a specific position in the core to fill the hydrophobic cavity on the protein surface, thereby enhancing binding affinity and selectivity. When the HDAC inhibitor SAHA is used in combination with the NAE inhibitor V11, it exhibits significant synergistic antiproliferative activity against hematologic malignancies (combination index CI = 0.475), providing direct experimental evidence for the pharmacological synergistic effect between the two targets. However, combination therapy has unavoidable inherent technical drawbacks in clinical treatment, namely, reduced patient medication adherence, complex pharmacokinetic behavior, and the risk of drug-drug interactions, which significantly limit its clinical application value. Summary of the Invention

[0005] This invention addresses the shortcomings of existing technologies by providing a class of small molecules containing indole and triazine groups and their application in the preparation of NAE / HDAC dual-target inhibitors.

[0006] In a first aspect, the present invention provides a class of small molecules containing indole and triazine groups, as shown in the following general formula:

[0007] ALB or BLA;

[0008] Wherein, group A is an indoleamino group or a derivative thereof, and the structure of group A is any one of the structures shown below. "Represents a connection site;

[0009] ;

[0010] The L group is a 1,2,4-triazine derivative, covalently linked to A and B, and the structure of the L group can be any one of the structures shown below. "Represents a connection site;

[0011] ;

[0012] Wherein, the B group is an N-hydroxyamide derivative that has substituted an aromatic group, and the structure of the B group is any one of the structures shown below. "Represents a connection site; ;

[0013] Wherein, n is an integer from 1 to 10; and preferably, the value of n is an integer from 4 to 9.

[0014] Furthermore, the preferred compounds of the present invention include Class Ia compounds, Class Ib compounds, Class Ic compounds, Class Id compounds, Class IIa compounds, and Class IIb compounds;

[0015] The structural formulas of the Class Ia compounds are shown below:

[0016] ;

[0017] The structural formulas of the Class Ib compounds are shown below, wherein R is selected from any one of -H, -F, -CF3, -OCH3, and -CH3:

[0018] ;

[0019] The structural formulas of the Ic class compounds are shown below:

[0020] ;

[0021] The structural formula of the Id class compounds is shown below:

[0022] ;

[0023] The structural formulas of the Class IIa compounds are shown below:

[0024] ;

[0025] The structural formulas of the Class IIb compounds are shown below:

[0026] ;

[0027] This invention also provides a method for preparing the Class Ia and IIa compounds, comprising the following preparation steps and preparation route:

[0028] .

[0029] The preparation steps of the Class Ia and IIa compounds are as follows: using p-nitrophenol and ethyl bromophenol as raw materials, tetrabutylammonium iodide as a phase transfer catalyst, and acetonitrile as a solvent, Williamson ether synthesis reaction is carried out to generate intermediate product 1. Then, intermediate product 1 is reduced with stannous chloride and concentrated hydrochloric acid as reducing conditions and anhydrous ethanol as a solvent to generate intermediate product 2. Using 3,5,6-trichloro-[1,2,4]-thiazide, 6-aminoindole, or 5-aminoindole as raw materials, triethylamine as an acid-binding agent, and tetrahydrofuran as a solvent, intermediate product 3 is generated. Then, intermediate product 3 and intermediate product 2 are reacted under camphor sulfonic acid catalysis to generate intermediate product 4. Finally, intermediate product 4 is reacted under alkaline conditions of NH2OK to generate the final product.

[0030] The present invention also provides a method for preparing the aforementioned Class Ib and Class IIb compounds, comprising the following preparation steps and preparation route;

[0031] .

[0032] The preparation steps of the Class Ib or Class IIb compounds are as follows: First, intermediate product 2 and 3,5,6-trichloro-[1,2,4]-thiazide are used as raw materials, triethylamine is used as an acid-binding agent, and tetrahydrofuran is used as a solvent to generate intermediate product 5. Then, intermediate product 5 and 6-aminoindole or 5-aminoindole are reacted under camphor sulfonic acid catalysis to generate intermediate product 6. Finally, intermediate product 6 is reacted under alkaline conditions of NH2OK to generate the final product.

[0033] The present invention also provides a method for preparing the aforementioned Class Ic compounds, comprising the following preparation steps and preparation route;

[0034] .

[0035] The preparation steps of the Class Ic compounds are as follows: First, using 6-nitropyridine-3-ol and ethyl bromoacetate as raw materials, tetrabutylammonium iodide as a phase transfer catalyst, and acetonitrile as a solvent, Williamson ether synthesis reaction is carried out to generate intermediate product 7. Then, intermediate product 7 is reduced with stannous chloride and concentrated hydrochloric acid as reducing conditions and anhydrous ethanol as a solvent to generate intermediate product 8. Using the above intermediate product 2 and 3,5,6-trichloro-[1,2,4]-thiazide as raw materials, triethylamine as an acid-binding agent, and tetrahydrofuran as a solvent, intermediate product 9 is generated. Then, intermediate product 5 and 6-aminoindole are reacted under camphor sulfonic acid catalysis to generate intermediate product 6. Finally, intermediate product 6 is reacted under alkaline conditions of NH2OK to generate the final product.

[0036] The present invention also provides a method for preparing the aforementioned Id class compounds, comprising the following preparation steps and preparation path;

[0037]

[0038] The preparation steps of the Id class compounds are as follows: First, using tert-butyl 1H-pyrazole-4-ylaminomethyl ester and ethyl bromomethyl ester as raw materials and acetonitrile as solvent, a nucleophilic substitution reaction is carried out to generate intermediate product 11. Then, intermediate product 11 is deprotected with dioxane hydrochloride to remove the Boc protecting group, and 3,5,6-trichloro-[1,2,4]-thiazide is added. Triethylamine is used as an acid-binding agent and tetrahydrofuran is used as a solvent to react and generate intermediate product 12. Then, intermediate product 12 and 6-aminoindole are reacted under camphor sulfonic acid catalysis to generate intermediate product 13. Finally, intermediate product 6 is reacted under alkaline conditions of NH2OK to generate the final product.

[0039] Secondly, the present invention provides the application of the above-mentioned derivative small molecules or their pharmaceutically acceptable salts, hydrates or prodrugs in the preparation of NAE / HDAC dual-target inhibitors, that is, to provide a novel NAE / HDAC dual-target inhibitor, wherein the NAE / HDAC dual-target inhibitor is the above-mentioned derivative small molecules or their pharmaceutically acceptable salts, hydrates or prodrugs, constructed using a pharmacophore fusion strategy.

[0040] It should be noted that "pharmaceutically acceptable" as used in this invention refers to any substance that does not interfere with the bioactivity of the active ingredient and is non-toxic to the host to which it is given. Unless otherwise stated, all tautomeric forms of the compounds of this invention are included within the scope of this invention. Furthermore, unless otherwise stated, the structural formulas of the compounds described in this invention include enriched isotopes of one or more different atoms.

[0041] Thirdly, the present invention provides the use of the above-mentioned derivative small molecules or their pharmaceutically acceptable salts, hydrates or prodrugs in the preparation of drugs for treating and / or preventing cardiovascular-related diseases, drugs for treating and / or preventing neurological-related diseases, drugs for treating and / or preventing immune system-related diseases, drugs for treating and / or preventing fibrotic diseases, and drugs for treating tumors. For example, the above-mentioned derivative small molecules or their pharmaceutically acceptable salts, hydrates or prodrugs, as NAE / HDAC dual-target inhibitors, can exert the synergistic effect of the pharmacophore, enhance the anti-tumor efficacy, and delay drug resistance caused by cross-activation between signaling pathways by interfering with multiple oncogenic / tumor-promoting pathways.

[0042] It should be noted that the aforementioned cardiovascular-related diseases include, but are not limited to, myocardial hypertrophy, heart failure, and atherosclerosis; the aforementioned nervous system-related diseases refer to neurodegenerative diseases, including, but not limited to, Alzheimer's disease and Huntington's disease; the aforementioned immune system-related diseases refer to autoimmune diseases, including, but not limited to, rheumatoid arthritis and inflammatory bowel disease; and the aforementioned fibrotic diseases include, but are not limited to, pulmonary fibrosis and liver fibrosis.

[0043] Beneficial effects: Through a rational pharmacophore fusion design strategy, the derivative small molecules developed in this invention serve as single-molecule multi-target inhibitors, which can overcome the inherent technical defects of combination therapy and retain synergistic inhibitory activity. Some derivative small molecules can even achieve inhibitory efficacy superior to existing NAE inhibitors and HDAC inhibitors in combination. Detailed Implementation

[0044] The examples provided below are merely illustrative of the invention and are not intended to limit the scope of the invention in any way. Unless otherwise specified, the raw materials may be obtained commercially or prepared by methods known in the art or according to the methods described herein. The HDAC positive drug SAHA (Vorinostat) was purchased from Shanghai Haohong Biomedical Technology Co., Ltd., and the NAE positive drug V11 was synthesized according to the synthetic route reported in the Experimental section of Appendix A. Supplementary data in the literature (Bioorg Med Chem Lett. 2020;30(2):126791).

[0045] The structures of the following compounds were determined by nuclear magnetic resonance (NMR). 1The determination was performed using 1H NMR and / or mass spectrometry (MS). NMR measurements were performed using a BRUKER AVANCE 300M or BRUKER AVANCE NEO 400M NMR spectrometer, with deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6) as the solvent and TMS as the internal standard. Column chromatography was performed using 200-300 mesh silica gel (manufactured by Shanghai Haohong Biomedical Technology Co., Ltd.). Preparative thin-layer chromatography was performed using silica gel plates (Silica Gel 60GF254, 1 mm coating thickness, sodium carboxymethyl cellulose binder, manufactured by Shanghai Haohong Biomedical Technology Co., Ltd.). Unless otherwise specified, all commercially available pharmaceutical products were purchased directly from Shanghai Haohong Biomedical Technology Co., Ltd., Shanghai Bide Pharmaceutical Technology Co., Ltd., and Anhui Zesheng Technology Co., Ltd. U-937 cells (human histiocytic lymphoma cells) were purchased from Suzhou Haixing Biotechnology Co., Ltd.

[0046] The reagents used in the biological laboratory are shown below:

[0047]

[0048] Example 1: 2-(4-((5-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)-N-hydroxyacetamide (Ia-1), with the following structural formula:

[0049] ;

[0050] The preparation process of Ia-1 is as follows:

[0051] Step S1, Preparation of N-(3,6-dichloro-1,2,4-triazine-5-yl)-1H-indole-6-amine: Compound 3,5,6-trichloro-[1,2,4]-thiazine (690 mg, 3.79 mmol), 6-aminoindole (500 mg, 3.79 mmol), triethylamine (459 mg, 4.55 mmol), and tetrahydrofuran (30 mL) were refluxed at 63 °C for 4 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained after concentration was purified by silica gel column chromatography (dichloromethane:methanol = 500:1) to give 950 mg of yellow solid (yield 90.0%). 1H NMR (300MHz, DMSO-d6) δ(ppm) 11.27 (s, 1H), 10.18 (s, 1H), 7.67 (s, 1H), 7.56 (d, J =8.5 Hz, 1H), 7.42 – 7.37 (m, 1H), 7.15 (dd, J = 8.6, 2.2 Hz, 1H), 6.51 – 6.39(m, 1H).ESI(m / z): 280.0153[M+H] + The structural formula of the obtained N-(3,6-dichloro-1,2,4-triazin-5-yl)-1H-indole-6-amine is as follows:

[0052] ;

[0053] Step S2, preparation of ethyl 2-(4-nitrophenoxy)acetic acid ester: A mixture of p-nitrophenol (1 g, 7.19 mmol), cesium carbonate (1.52 g, 7.9 mmol), tetrabutylammonium iodide (0.27 g, 0.72 mmol), and acetonitrile (30 mL) was stirred at room temperature for 0.5 h, followed by the addition of ethyl bromoacetate (1.20 g, 7.19 mmol). The reaction was carried out at 85 °C for 8 h. The reaction was monitored by TLC until completion. Insoluble matter was removed by diatomaceous earth filtration. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 50:1) to obtain 1.4 g of yellow solid (yield 87%). The structural formula of the obtained ethyl 2-(4-nitrophenoxy)acetic acid ester is as follows:

[0054] ;

[0055] Step S3, Preparation of Ethyl 2-(4-aminophenoxy)acetic acid: Under a nitrogen atmosphere, concentrated hydrochloric acid (5 mL) and anhydrous ethanol (25 mL) were added to ethyl 2-(4-nitrophenoxy)acetic acid (1.4 g, 6.2 mmol) and stannous chloride (8.18 g, 43.14 mmol), and the reaction was continued with stirring at room temperature for 12 h. The pH was adjusted to neutral using saturated sodium bicarbonate solution, and the insoluble matter was filtered off. The filtrate was extracted with dichloromethane (3 × 50 mL), and the extracts were combined, washed with water (3 × 60 mL), and dried over anhydrous sodium sulfate. After filtration, the residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 20:1) to give 0.8 g of brown solid (yield 67%). ESI (m / z): 196.0974 [M+H] + The structural formula of the obtained ethyl 2-(4-aminophenoxy)acetic acid ester is as follows:

[0056] ;

[0057] Step S4, preparation of ethyl 2-(4-((5-((1H-indole-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)acetate: Ethyl N-(3,6-dichloro-1,2,4-triazin-5-yl)-1H-indole-6-amine (107 mg, 0.547 mmol), ethyl 2-(4-aminophenoxy)acetate (153 mg, 0.548 mmol), camphor sulfonic acid (51 mg, 0.22 mmol), and isopropanol (30 mL) were refluxed at 85 °C for 12 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained from the concentration was purified by thin-layer chromatography (dichloromethane:methanol = 100:1) to give 126 mg of yellow solid (yield 54%). 1 H NMR (400MHz, DMSO-d6) δ(ppm) 11.18 (s, 1H), 9.46 (s, 1H), 9.45 (s, br, 1H), 7.58 (s,1H), 7.55 (d, J = 8.7 Hz, 1H), 7.47 (d, 2H), 7.40 – 7.35 (m, 1H), 7.20 (dd, J= 8.5, 1.9 Hz, 1H), 6.69 (d, J = 9.1 Hz, 2H), 6.48 – 6.40 (m, 1H), 4.66 (s,2H), 4.16 (q, J = 7.1 Hz, 2H), 1.21 (d, J = 7.1 Hz, 3H). ESI (m / z): 439.1267 [M+H] + The structural formula of the prepared ethyl 2-(4-((5-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazine-3-yl)amino)phenoxy)acetate is as follows:

[0058] ;

[0059] Step S5, preparation of 2-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)-N-hydroxyacetamide (Ia-1): A methanol solution of potassium hydroxide (8.4 g, 150.1 mmol) was placed in an ice bath, and then a methanol solution of hydroxylamine hydrochloride (7.0 g, 100.6 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 30 min. Insoluble matter was removed by filtration to obtain an NH2OK / methanol solution. Ethyl 2-(4-((5-(((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)acetate (39 mg, 0.091 mmol) was added to 8 mL of the NH2OK / methanol solution, and stirring was continued for 1 h. After the reaction was completed by TLC monitoring, the pH of the reaction mixture was adjusted to 6–7 with 1M hydrochloric acid. The reaction solution was concentrated by vacuum distillation, and the residue was washed with water to give 30 mg of yellow solid (yield 77%). 1 H NMR (300 MHz, DMSO-d6) δ(ppm)11.15 (s, 1H), 10.79 (s, 1H), 9.43 (s, 1H), 8.95 (s, 1H), 7.58 (d, J = 1.7Hz, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.48 (d, J = 9.0 Hz, 2H), 7.40 – 7.35 (m,1H), 7.20 (dd, J = 8.5, 1.9 Hz, 1H), 6.74 (d, J = 9.1 Hz, 2H), 6.48 – 6.42(m, 1H), 4.35 (s, 2H). ESI(m / z): 426.1087 [M+H] + .

[0060] Example 2: 4-(4-((5-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)-N-hydroxybutyramide (Ia-2), with the following structural formula:

[0061] ;

[0062] The only difference between the synthesis steps of Example 2 and Example 1 is that in step S4, the ethyl 2-(4-amino) 1 The remaining steps are the same; (HNMR) phenoxy) acetate is substituted for ethyl 2-(4-aminophenoxy) butyrate. 1H NMR (400MHz, DMSO-d6) δ(ppm) 11.17 (s, 1H), 10.42 (s, 1H), 9.43 (s, 1H), 9.40 (s,1H), 8.72 (s, 1H), 7.57 (s, 1H), 7.55 (d, J = 8.7 Hz, 1H), 7.46 (d, J = 8.6Hz, 2H), 7.40 – 7.36 (m, 1H), 7.19 (dd, J = 8.4, 1.8 Hz, 1H), 6.68 (d, J =8.6 Hz, 2H), 6.49 – 6.42 (m, 1H), 3.84 (t, J = 6.3 Hz, 2H), 2.10 (t, J = 7.4Hz, 2H), 1.93 – 1.85 (m, 2H). ESI(m / z): 454.1295[M+H] +

[0063] Example 3: 5-(4-((5-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)-N-hydroxypentanamide (Ia-3), with the following structural formula:

[0064] ;

[0065] The difference between the synthesis steps of Example 3 and Example 1 is that in step S4, ethyl 2-(4-aminophenoxy)acetic acid ester is replaced with ethyl 2-(4-aminophenoxy)valerate. 1 H NMR (400 MHz, DMSO-d6) δ(ppm)11.19 (s, 1H), 10.41 (s, 1H), 9.44 (s, 1H), 9.41 (s, 1H), 8.74 (s, 1H), 7.59(s, 1H), 7.56 (d, J = 8.3 Hz, 1H), 7.47 (d, J = 8.4 Hz, 2H), 7.41 – 7.35 (m,1H), 7.25 – 7.11 (m, 1H), 6.69 (d, J = 8.5 Hz, 2H), 6.52 – 6.39 (m, 1H), 3.86(d, J = 6.4 Hz, 2H), 2.01 (t, J = 6.4 Hz, 2H), 1.75 – 1.53 (m, 4H). ESI(m / z):468.1555[M+H] +

[0066] Example 4: 6-(4-((5-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)-N-hydroxyhexanoamide (Ia-4), with the following structural formula:

[0067] ;

[0068] The difference between the synthesis steps of Example 4 and Example 1 is that in step S4, ethyl 2-(4-aminophenoxy)acetic acid ester is replaced with ethyl 2-(4-aminophenoxy)hexanoate ester. 1 H NMR (400 MHz, DMSO-d6) δ(ppm)11.19 (s, 1H), 10.38 (s, 1H), 9.44 (s, 1H), 9.41 (s, 1H), 8.72 (s, 1H), 7.58(s, 1H), 7.56 (d, J = 8.3 Hz, 1H), 7.46 (d, J = 8.5 Hz, 2H), 7.41 – 7.35 (m,1H), 7.19 (d, J = 8.4 Hz, 1H), 6.68 (d, J = 8.4 Hz, 2H), 6.50 – 6.43 (m, 1H), 3.83 (t, J = 6.3 Hz, 2H), 1.98 (t, J = 7.3 Hz, 2H), 1.75 – 1.60 (m, 2H), 1.59 – 1.46 (m, 2H), 1.42 – 1.29 (m, 2H).ESI(m / z):482.1710[M+H] +

[0069] Example 5: 7-(4-((5-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)-N-hydroxyheptanamide (Ia-5), with the following structural formula:

[0070] ;

[0071] The difference between the synthesis steps of Example 5 and Example 1 is that in step S4, ethyl 2-(4-aminophenoxy)acetic acid ester is replaced with ethyl 2-(4-aminophenoxy)heptanoate ester. 1H NMR (400 MHz, DMSO-d6) δ(ppm)11.20 (s, 1H), 10.37 (s, 1H), 9.44 (s, 1H), 9.39 (s, br, 1H), 8.68 (s, 1H),7.58 (s, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.45 (d, J = 8.5 Hz, 2H), 7.40 – 7.35(m, 1H), 7.18 (dd, J = 8.4, 1.8 Hz, 1H), 6.68 (d, J = 8.5 Hz, 2H), 6.48 –6.43 (m, 1H), 3.84 (t, J = 6.5 Hz, 2H), 1.95 (t, J = 7.3 Hz, 2H), 1.71 – 1.59(m, 2H), 1.56 – 1.45 (m, 2H), 1.42 – 1.33 (m, 2H), 1.32 – 1.25 (m, 2H). ESI(m / z): 496.1868[M+H] +

[0072] Example 6: 8-(4-((5-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)-N-hydroxyoctamide (Ia-6), with the following structural formula:

[0073] ;

[0074] The difference between the synthesis steps of Example 6 and Example 1 is that in step S4, ethyl 2-(4-aminophenoxy)acetic acid ester is replaced with ethyl 2-(4-aminophenoxy)octanoate. 1H NMR (400 MHz, DMSO-d6) δ(ppm)11.18 (s, 1H), 10.35 (s, 1H), 9.43 (s, 1H), 9.40 (s, 1H), 8.69 (s, 1H), 7.58(s, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.45 (d, J = 8.6 Hz, 2H), 7.39 – 7.35 (m,1H), 7.19 (dd, J = 8.3, 1.8 Hz, 1H), 6.68 (d, J = 8.5 Hz, 2H), 6.48 – 6.42(m, 1H), 3.84 (t, J = 6.5 Hz, 2H), 1.95 (t, J = 7.3 Hz, 2H), 1.69 – 1.60 (m,2H), 1.54 – 1.46 (m, 2H), 1.41 – 1.17 (m, 8H). ESI(m / z): 510.2005[M+H] +

[0075] Example 7: 10-(4-((5-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)-N-hydroxydecanoamide (Ia-7), with the following structural formula:

[0076] ;

[0077] The difference between the synthesis steps in Example 7 and those in Example 1 is that in step S4, ethyl 2-(4-aminophenoxy)acetic acid ester is replaced with ethyl 2-(4-aminophenoxy)decanoate ester. 1H NMR (400 MHz, DMSO-d6) δ(ppm)11.18 (s, 1H), 10.33 (s, 1H), 9.43 (s, 1H), 9.39 (s, br, 1H), 8.67 (s, 1H),7.57 (s, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.45 (d, J = 8.6 Hz, 2H), 7.38 – 7.32(m, 1H), 7.18 (dd, J = 8.5, 1.8 Hz, 1H), 6.67 (d, J = 8.6 Hz, 2H), 6.47 –6.41 (m, 1H), 3.84 (t, J = 6.5 Hz, 2H), 1.93 (t, J = 7.4 Hz, 2H), 1.69 – 1.60(m, 2H), 1.52 – 1.43 (m, 2H), 1.41 – 1.33 (m, 2H), 1.31 – 1.23 (m, 8H). ESI(m / z): 538.2336[M+H] +

[0078] Example 8: 8-(4-((5-((1H-indol-5-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)-N-hydroxyoctamide (IIa-1), with the following structural formula:

[0079] ;

[0080] Step S1, Preparation of N-(3,6-dichloro-1,2,4-triazine-5-yl)-1H-indole-5-amine: Compound 3,5,6-trichloro-[1,2,4]-thiazine (690 mg, 3.79 mmol), 5-aminoindole (500 mg, 3.79 mmol), triethylamine (459 mg, 4.55 mmol), and tetrahydrofuran (30 mL) were refluxed at 63 °C for 4 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained after concentration was purified by silica gel column chromatography (dichloromethane:methanol = 500:1) to give 950 mg of yellow solid (yield 90.0%). 1H NMR(400 MHz, DMSO-d6) δ(ppm) 11.25 (s, 1H), 10.21 (s, 1H), 7.67 (s, 1H), 7.44(d, J = 8.6 Hz, 1H), 7.42 – 7.39 (m, 1H), 7.20 (dd, J = 8.7, 2.0 Hz, 1H),6.53 – 6.41 (m, 1H). ESI(m / z): 280.0158[M+H] + The product N-(3,6-dichloro-1,2,4-triazin-5-yl)-1H-indole-5-amine has the following structural formula:

[0081] ;

[0082] The remaining synthesis steps S2-S3 are the same as steps S2-S3 in Example 1;

[0083] Step S4 differs from Step S4 in Example 1 only in that N-(3,6-dichloro-1,2,4-triazin-5-yl)-1H-indole-5-amine is used instead of N-(3,6-dichloro-1,2,4-triazin-5-yl)-1H-indole-6-amine to prepare ethyl 8-(4-((5-((1H-indole-5-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)octanoate. 1 HNMR (400 MHz, DMSO-d6) δ(ppm) 11.16 (s, 1H), 9.40 (s, br, 1H), 9.36 (s, 1H), 7.81 (s, 1H), 7.46 (d, J = 8.6 Hz, 2H), 7.42 (d, J = 8.7 Hz, 1H), 7.40 – 7.37(m, 1H), 7.25 (dd, J = 8.7, 2.1 Hz, 1H), 6.69 (d, J = 8.6 Hz, 2H), 6.47 –6.41 (m, 1H), 4.05 (q, J = 7.1 Hz, 2H), 3.85 (t, J = 6.5 Hz, 2H), 1.72 – The structural formula of ethyl 8-(4-((5-((1H-indol-5-yl)amino)-6-chloro-1,2,4-triazin-3-yl)amino)phenoxy)octanoate is as follows: (T, J = 7.1 Hz, 3H)

[0084] ;

[0085] Step S5: Prepare the final product 8-(4-((5-((1H-indol-5-yl)amino)-6-chloro-1,2,4-triazine-3-yl)amino)phenoxy)-N-hydroxyoctamide (IIa-1), the preparation steps are the same as step S5 in Example 1; 1 H NMR(300 MHz, DMSO-d6) δ(ppm) 11.16 (s, 1H), 10.36 (s, br, 1H), 9.38 (s, 1H),9.35 (s, br, 1H), 8.70 (s, br, 1H), 7.80 (s, 1H), 7.45 (d, J = 8.3 Hz, 2H), 7.41 (d, J = 8.1 Hz, 1H), 7.40 – 7.37 (m, 1H), 7.24 (dd, J = 8.6, 2.0 Hz, 1H), 6.69 (d, J = 8.9 Hz, 2H), 6.48 – 6.40 (m, 1H), 3.86 (t, J = 6.4 Hz, 2H),1.95 (t, J = 7.3 Hz, 2H), 1.73 – 1.59 (m, 2H), 1.55 – 1.44 (m, 2H), 1.41 –1.25 (m, 6H). ESI(m / z): 510.2041[M+H] +

[0086] Example 9: 2-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)-N-hydroxyacetamide (Ib-1), with the following structural formula:

[0087] ;

[0088] Step S1, preparation of ethyl 2-(4-nitrophenoxy)acetic acid ester: A mixture of p-nitrophenol (1 g, 7.19 mmol), cesium carbonate (1.52 g, 7.9 mmol), tetrabutylammonium iodide (0.27 g, 0.72 mmol), and acetonitrile (30 mL) was stirred at room temperature for 0.5 h, followed by the addition of ethyl bromoacetate (1.20 g, 7.19 mmol). The reaction was carried out at 85 °C for 8 h. The reaction was monitored by TLC until completion. Insoluble matter was removed by diatomaceous earth filtration. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1) to obtain 1.4 g of yellow solid (yield 87%). The structural formula of the prepared ethyl 2-(4-nitrophenoxy)acetic acid ester is as follows:

[0089] ;

[0090] Step S2, Preparation of ethyl 2-(4-aminophenoxy)acetic acid: Under a nitrogen atmosphere, concentrated hydrochloric acid (5 mL) and anhydrous ethanol (25 mL) were added to ethyl 2-(4-nitrophenoxy)acetic acid (1.4 g, 6.2 mmol) and stannous chloride (8.18 g, 43.14 mmol), and the reaction was continued to be stirred at room temperature for 12 h. The pH was adjusted to neutral using saturated sodium bicarbonate solution, and the insoluble matter was filtered off. The filtrate was extracted with dichloromethane (3 × 50 mL), and the extracts were combined, washed with water (3 × 60 mL), and dried over anhydrous sodium sulfate. After filtration, the residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to give 0.8 g of brown solid (yield 67%); ESI (m / z): 196.0974 [M+H]+; The structural formula of the prepared ethyl 2-(4-aminophenoxy)acetic acid is as follows:

[0091] ;

[0092] Step S3, preparation of ethyl 2-(4-((3,6-dichloro-1,2,4-triazine-5-amino)phenoxy)acetate: Compound 3,5,6-trichloro-[1,2,4]-thiazine (0.246 mg, 1.35 mmol), ethyl 2-(4-aminophenoxy)acetate (0.264 mg, 1.35 mmol), triethylamine (160 mg, 1.62 mmol), and tetrahydrofuran (30 mL) were refluxed at 63 °C for 4 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained from the concentration was purified by silica gel column chromatography (dichloromethane:methanol = 500:1) to give 0.40 g of yellow solid (yield 86.1%). 1H NMR (400 MHz, CDCl3) δ (ppm) 7.84 (s, 1H), 7.55 –7.48 (m, 2H), 6.96 – 6.88 (m, 2H), 4.62 (s, 2H), 4.26 (q, J = 7.1, 2H), 1.29(t, J = 7.1, 3H). ESI(m / z):343.0362[M+H] + The structural formula of the prepared ethyl 2-(4-((3,6-dichloro-1,2,4-triazine-5-amino)phenoxy)acetate is as follows:

[0093] ;

[0094] Step S4, preparation of ethyl 2-(4-((3-((1H-indole-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)acetate: Ethyl 2-(4-((3,6-dichloro-1,2,4-triazin-5-amino)phenoxy)acetate (150 mg, 0.44 mmol), 6-aminoindole (58 mg, 0.44 mmol), camphor sulfonic acid (42 mg, 0.18 mmol), and isopropanol (30 mL) were refluxed at 85 °C for 12 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained from the concentration was purified by thin-layer chromatography (dichloromethane:methanol = 100:1) to give 72 mg of yellow solid (yield 37%). 1 H NMR (400 MHz, CDCl3) δ(ppm)8.50 (s, 1H), 7.92 (s, 1H), 7.51 (d, J = 8.0 Hz, 1H), 7.48 (d, J = 8.8 Hz,2H), 7.40 (s, br, 1H), 7.19 – 7.13 (m, 2H), 6.97 (d, J = 8.5 Hz, 2H), 6.82 (dd, J = 8.4, 2.0 Hz, 1H), 6.50 – 6.44 (m, 1H), 4.72 (s, 2H), 4.30 (q, J =7.1 Hz, 2H), 1.32 (d, J = 3.7 Hz, 3H). ESI(m / z): 439.1287[M+H] + The structural formula of the prepared ethyl 2-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)acetate is as follows:

[0095] ;

[0096] Step S5, preparation of 2-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)-N-hydroxyacetamide (Ib-1): A methanol solution of potassium hydroxide (8.4 g, 150.1 mmol) was placed in an ice bath, and then a methanol solution of hydroxylamine hydrochloride (7.0 g, 100.6 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 30 min. Insoluble matter was removed by filtration to obtain an NH2OK / methanol solution. Ethyl 2-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)acetate (35 mg, 0.078 mmol) was added to 8 mL of the NH2OK / methanol solution, and stirring was continued for 1 h. After the reaction was completed by TLC monitoring, the pH of the reaction mixture was adjusted to 6–7 with acetic acid. The reaction solution was concentrated by vacuum distillation, and the residue was washed with water to give 27 mg of yellow solid (yield 81%). 1 H NMR (300 MHz, DMSO-d6) δ(ppm)10.88 (s, 1H), 10.86 (s, 1H), 9.44 (s, 1H), 9.27 (s, 1H), 9.02 (s, 1H), 7.80(s, 1H), 7.63 (d, J = 8.7 Hz, 2H), 7.38 (d, J = 8.5 Hz, 1H), 7.26 – 7.21 (m,1H), 7.19 (dd, J = 8.6, 1.9 Hz, 1H), 6.97 (d, J = 8.9 Hz, 2H), 6.39 – 6.30(m, 1H), 4.48 (s, 2H). ESI (m / z): 426.1083 [M+H] + .

[0097] Example 10: 4-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)-N-hydroxybutyramide (Ib-2), with the following structural formula:

[0098] ;

[0099] The difference between the synthesis steps of Example 10 and Example 9 is that in step S3, 2-(4-aminophenoxybutyrate is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester; 1H NMR (400 MHz, DMSO-d6) δ(ppm) 10.92 (s,1H), 10.45 (s, 1H), 9.41 (s, 1H), 9.25 (s, 1H), 8.74 (s, 1H), 7.79 (s, 1H),7.61 (d, J = 8.9 Hz, 2H), 7.38 (d, J = 8.6 Hz, 1H), 7.27 – 7.21 (m, 1H), 7.18 (dd, J = 8.5, 1.8 Hz, 1H), 6.93 (d, J = 9.1 Hz, 3H), 6.37 – 6.32 (m, 1H), 3.96 (t, J = 6.3 Hz, 3H), 2.14 (t, J = 7.4 Hz, 2H), 1.96 – 1.90 (m, 2H). ESI(m / z): 454.1401[M+H] + Example 11: 5-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazine-5-yl)amino)phenoxy)-N-hydroxypentanamide (Ib-3), with the following structural formula:

[0100] ;

[0101] The only difference between the synthesis steps of Example 11 and Example 9 is that in step S3, 2-(4-aminophenoxyvalerate) ester is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester. 1 H NMR (400 MHz, DMSO-d6) δ(ppm) 10.92 (s,1H), 10.41 (s, 1H), 9.41 (s, 1H), 9.25 (s, 1H), 8.73 (s, 1H), 7.78 (s, 1H),7.60 (d, J = 8.9 Hz, 2H), 7.38 (d, J = 8.5 Hz, 1H), 7.26 – 7.22 (m, 1H), 7.18 (dd, J = 8.5, 1.9 Hz, 1H), 6.94 (d, J = 8.9 Hz, 2H), 6.39 – 6.31 (m, 1H), 4.00 – 3.93 (m, 2H), 2.03 (t, J = 6.8 Hz, 2H), 1.77 – 1.62 (m, 4H). ESI(m / z):468.1557[M+H] + .

[0102] Example 12: 6-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazine-5-yl)amino)phenoxy)-N-hydroxyhexanoamide (Ib-4), with the following structural formula:

[0103] ;

[0104] The only difference between the synthesis steps of Example 12 and Example 9 is that in step S3, 2-(4-aminophenoxyhexanoate is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester. 1 H NMR (400 MHz, DMSO-d6) δ(ppm) 10.92 (s,1H), 10.37 (s, 1H), 9.41 (s, 1H), 9.24 (s, 1H), 8.70 (s, 1H), 7.77 (s, 1H),7.59 (d, J = 8.6 Hz, 3.94 (t, J = 6.4 Hz, 2H), 1.98 (t, J = 7.3 Hz, 2H), 1.78 – 1.64 (m, 2H), 1.60– 1.51 (m, 2H), 1.47 – 1.35 (m, 2H). ESI(m / z): 482.1726[M+H] + .

[0105] Example 13: 7-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)-N-hydroxyheptanamide (Ib-5), with the following structural formula:

[0106] ;

[0107] The only difference between the synthesis steps of Example 13 and Example 9 is that in step S3, 2-(4-aminophenoxyheptanoate ester is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester. 1H NMR (400 MHz, DMSO-d6) δ(ppm) 10.91 (s,1H), 10.39 (s, 1H), 9.41 (s, 1H), 9.25 (s, 1H), 8.72 (s, 1H), 7.77 (s, 1H),7.59 (d, J = 8.4 Hz, 2H), 7.38 (d, J = 8.5 Hz, 1H), 7.26 – 7.20 (m, 1H), 7.18(d, J = 8.5 Hz, 1H), 6.92 (d, J = 8.4 Hz, 2H), 6.43 – 6.25 (m, 1H), 3.94 (t,J = 6.4 Hz, 2H), 1.97 (t, J = 7.3 Hz, 2H), 1.77 – 1.61 (m, 2H), 1.58 – 1.46(m, 2H), 1.44 – 1.36 (m, 2H), 1.35 – 1.21 (m, 2H). ESI(m / z):496.1870[M+H] + .

[0108] Example 14: 8-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazine-5-yl)amino)phenoxy)-N-hydroxyoctamide (Ib-6), with the following structural formula:

[0109] ;

[0110] The only difference between the synthesis steps of Example 14 and Example 9 is that in step S3, 2-(4-aminophenoxyoctanoate is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester. 1H NMR (400 MHz, DMSO-d6) δ(ppm) 10.93 (s,1H), 10.39 (s, br, 1H), 9.43 (s, 1H), 9.26 (s, br, 1H), 8.76 (s, br, 1H),7.79 (s, 1H), 7.60 (d, J = 8.4 Hz, 2H), 7.39 (d, J = 8.5 Hz, 1H), 7.25 – 7.22(m, 1H), 7.20 (d, J = 8.6 Hz, 1H), 6.92 (d, J = 8.4 Hz, 2H), 6.38 – 6.31 (m,1H), 3.93 (s, 2H), 1.97 (t, J = 7.3 Hz, 2H), 1.76 – 1.63 (m, 2H), 1.57 – 1.46(m, 2H), 1.38 – 1.18 (m, 6H). ESI(m / z): 510.2037[M+H] + .

[0111] Example 15: 10-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)-N-hydroxydecanoamide (Ib-7), with the following structural formula:

[0112] ;

[0113] The only difference between the synthesis steps of Example 15 and Example 9 is that in step S3, 2-(4-aminophenoxydecanoate is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester. 1H NMR (300 MHz, DMSO-d6) δ(ppm) 10.92 (s,1H), 10.34 (s, br, 1H), 9.42 (s, 1H), 9.25 (s, br, 1H), 8.68 (s, br, 1H),7.77 (s, 1H), 7.59 (d, J = 8.7 Hz, 2H), 7.37 (d, J = 8.5 Hz, 1H), 7.29 – 7.20(m, 1H), 7.18 (dd, J = 8.5, 1.9 Hz, 1H), 6.93 (d, J = 8.7 Hz, 2H), 6.38 –6.29 (m, 1H), 3.95 (t, J = 6.5 Hz, 2H), 1.94 (t, J = 7.3 Hz, 2H), 1.76 – 1.66(m, 2H), 1.54 – 1.27 (m, 12H). ESI(m / z): 538.2342[M+H] + .

[0114] Example 16: 7-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-2-fluorophenoxy)-N-hydroxyheptanamide (Ib-8), with the following structural formula:

[0115] ;

[0116] The difference between the synthesis steps of Example 16 and Example 9 is that in step S3, ethyl 7-(4-amino-2-fluorophenoxy)heptanoate is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester. 1H NMR (300 MHz, DMSO-d6) δ(ppm)10.94 (s, 1H), 10.36 (s, br, 1H), 9.51 (s, 1H), 9.33 (s, br, 1H), 8.71 (s,br, 1H), 7.78 (s, 1H), 7.72 (d, J = 11.8 Hz, 1H), 7.48 (d, J = 9.0 Hz, 1H), 7.40 (d, J = 8.5 Hz, 1H), 7.23 (s, 1H), 7.20 (d, J = 7.5 Hz, 1H), 7.18 – 7.03(m, 1H), 6.56 – 6.14 (m, 1H), 4.02 (t, J = 6.4 Hz, 2H), 1.96 (t, J = 7.3 Hz,2H), 1.82 – 1.62 (m, 2H), 1.61 – 1.47 (m, 2H), 1.45 – 1.35 (m, 2H), 1.36 –1.27 (m, 2H). ESI(m / z): 514.1766[M+H] + .

[0117] Example 17: 7-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-2-methylphenoxy)-N-hydroxyheptanamide (Ib-9), with the following structural formula:

[0118] ;

[0119] The difference between the synthesis steps in Example 17 and those in Example 9 is that in step S3, ethyl 7-(4-amino-2-trifluoromethylphenoxy) is substituted for ethyl 2-(4-aminophenoxy) acetate. 1H NMR (300 MHz, DMSO-d6) δ(ppm)10.96 (s, 1H), 10.38 (s, br, 1H), 9.51 (s, 1H), 9.33 (s, br, 1H), 8.73 (s,br, 1H), 8.03 (dd, J = 9.1, 2.6 Hz, 1H), 7.84 (d, J = 2.5 Hz, 1H), 7.74 (s,1H), 7.38 (d, J = 8.5 Hz, 1H), 7.26 – 7.23 (m, 1H), 7.22 (d, J = 2.6 Hz, 1H),7.22 – 7.17 (m, 1H), 6.51 – 6.15 (m, 1H), 4.10 (t, J = 6.2 Hz, 2H), 1.97 (t,J = 7.3 Hz, 2H), 1.83 – 1.64 (m, 2H), 1.60 – 1.49 (m, 2H), 1.48 – 1.38 (m,2H), 1.38 – 1.27 (m, 2H). ESI(m / z):564.1709[M+H] + .

[0120] Example 18: 7-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-2-methylphenoxy)-N-hydroxyheptanamide (Ib-10), with the following structural formula:

[0121] ;

[0122] The difference between the synthesis steps of Example 18 and Example 9 is that in step S3, ethyl 7-(4-amino-2-methoxyphenoxy)heptanoate is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester. 1H NMR (300 MHz, DMSO-d6) δ10.90 (s, 1H), 10.37 (s, 1H), 9.44 (s, 1H), 9.22 (s, 1H), 8.70 (s, 1H), 7.79(s, 1H), 7.37 (d, J = 8.5 Hz, 1H), 7.34 – 7.30 (m, 1H), 7.29 (s, 1H), 7.26 –7.22 (m, 1H), 7.22 – 7.18 (m, 1H), 6.93 (d, J = 8.5 Hz, 1H), 6.38 – 6.32 (m,1H), 3.94 (t, J = 6.5 Hz, 2H), 3.67 (s, 3H), 1.97 (t, J = 7.3 Hz, 2H), 1.79 –1.63 (m, 2H), 1.58 – 1.46 (m, 2H), 1.45 – 1.36 (m, 2H), 1.34 – 1.26 (m, 2H).ESI(m / z):526.1957[M+H] + .

[0123] Example 19: 7-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-2-methylphenoxy)-N-hydroxyheptanamide (Ib-11), with the following structural formula:

[0124] ;

[0125] The difference between the synthesis steps of Example 19 and Example 9 is that in step S3, ethyl 7-(4-amino-2-methylphenoxy)heptanoate is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester. 1H NMR (300 MHz, DMSO-d6) δ(ppm) 10.91 (s, 1H), 10.36 (s, 1H), 9.41 (s, 1H), 9.16 (s, 1H), 8.68 (s, 1H),7.73 (s, 1H), 7.52 (d, J = 15.0 Hz, 1H), 7.46 – 7.43 (m, 1H), 7.38 (d, J =8.6 Hz, 1H), 7.24 – 7.22 (m, 1H), 7.21 – 7.18 (m, 1H), 6.91 (d, J = 9.4 Hz,1H), 6.38 – 6.28 (m, 1H), 3.95 (t, J = 6.3 Hz, 2H), 2.09 (s, 3H), 1.96 (t, J= 7.3 Hz, 2H), 1.79 – 1.67 (m, 2H), 1.59 – 1.49 (m, 2H), 1.48 – 1.40 (m, 2H), 1.32 (d, J = 7.3 Hz, 2H). ESI(m / z): 510.2020[M+H] + .

[0126] Example 20: 7-((6-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)-N-hydroxyheptanamide (Ic-1), with the following structural formula:

[0127] ;

[0128] Step S1: Preparation of ethyl 7-((6-nitropyridin-3-yl)oxy)heptanoate: A mixture of 6-nitropyridin-3-ol (0.5 g, 3.57 mmol), cesium carbonate (1.28 g, 3.93 mmol), tetrabutylammonium iodide (0.26 g, 0.70 mmol), and acetonitrile (30 mL) was stirred at room temperature for 0.5 h, followed by the addition of ethyl bromoheptanoate (1.27 g, 5.36 mmol). The reaction was carried out at 85 °C for 8 h. The reaction was monitored by TLC until completion. Insoluble matter was removed by diatomaceous earth filtration. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 50:1) to give 0.97 g of yellow solid (yield 92%). The structural formula of the prepared ethyl 7-((6-nitropyridin-3-yl)oxy)heptanoate is as follows:

[0129] ;

[0130] Step S2, Preparation of Ethyl 7-((6-aminopyridin-3-yl)oxy)heptanoate: Under a nitrogen atmosphere, concentrated hydrochloric acid (5 mL) and anhydrous ethanol (25 mL) were added to ethyl 2-(4-nitrophenoxy)acetate (0.94 g, 3.17 mmol) and stannous chloride (3.7 g, 19.0 mmol), and the reaction was continued to be stirred at room temperature for 12 h. The pH was adjusted to neutral using saturated sodium bicarbonate solution, and the insoluble matter was filtered off. The filtrate was extracted with dichloromethane (3 × 50 mL), and the extracts were combined, washed with water (3 × 60 mL), and dried over anhydrous sodium sulfate. After filtration, the residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to give 0.54 g of green solid (yield 64%). The structural formula of the prepared ethyl 7-((6-aminopyridin-3-yl)oxy)heptanoate is as follows:

[0131] ;

[0132] Step S3, preparation of ethyl 7-((6-((3,6-dichloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)heptanoate: Compound 3,5,6-trichloro-[1,2,4]-thiazine (0.200 g, 1.10 mmol), ethyl 2-(4-((3,6-dichloro-1,2,4-triazin-5-yl)phenoxy)acetate (0.293 g, 1.10 mmol), triethylamine (133 mg, 1.32 mmol), and tetrahydrofuran (30 mL) were refluxed at 63 °C for 4 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained from the concentration was purified by thin-layer chromatography (dichloromethane:methanol = 95:1) to give 0.32 g of yellow solid (yield 71.0%). 1 H NMR (300 MHz, DMSO-d6) δ(ppm) 10.46 (s, 1H), 8.18 (d, J = 3.0 Hz, 1H), 7.69 (d, J = 8.9 Hz, 1H), 7.56 (dd, J = 9.0, 3.0 Hz, 1H), 4.10 – 3.99 (m, 4H), 2.29 (t, J = 7.3Hz, 2H), 1.81 – 1.66 (m, 2H), 1.63 – 1.49 (m, 2H), 1.48 – 1.28 (m, 4H), 1.17(t, J = 7.1 Hz, 3H). The structural formula for preparing ethyl 7-((6-((3,6-dichloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)heptanoate is as follows:

[0133] ;

[0134] Step S4, preparation of ethyl 7-((6-((3-((1H-indole-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)heptanoate: Ethyl 7-((6-((3,6-dichloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)heptanoate (83 mg, 0.2 mmol), 6-aminoindole (27 mg, 0.20 mmol), camphorsulfonic acid (19 mg, 0.08 mmol) and isopropanol (30 mL) were refluxed at 85 °C for 12 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained from the concentration was purified by thin-layer chromatography (dichloromethane:methanol = 100:1) to give 40 mg of a yellow solid (yield 39.8%). The structural formula of the prepared 7-((6-((3,6-dichloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)heptanoate is as follows:

[0135] ;

[0136] Step S5, preparation of ((6-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)-N-hydroxyheptanamide (Ic-1): A methanol solution of potassium hydroxide (8.4 g, 150.1 mmol) was placed in an ice bath, and then a methanol solution of hydroxylamine hydrochloride (7.0 g, 100.6 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 30 min. Insoluble matter was removed by filtration to obtain an NH2OK / methanol solution. 7-((6-((3,6-dichloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)heptanamide (40 mg, 0.080 mmol) was added to 8 mL of the NH2OK / methanol solution, and stirring was continued for 1 h. After the reaction was completed by TLC monitoring, the pH of the reaction mixture was adjusted to 6–7 with acetic acid. The reaction solution was concentrated by vacuum distillation, and the residue was washed with water to give 26 mg of yellow solid (65% yield). 1H NMR (400 MHz, DMSO-d6) δ(ppm) 10.98(s, 1H), 10.37 (s, 1H), 9.59 (s, 1H), 9.22 (s, 1H), 8.70 (s, 1H), 8.16 (d, J= 3.0 Hz, 1H), 8.02 (d, J = 8.9 Hz, 1H), 7.82 (s, 1H), 7.43 (d, J = 8.8 Hz,1H), 7.41 (d, J = 8.4 Hz, 1H), 7.28 – 7.22 (m, 1H), 7.17 (dd, J = 8.5, 1.9Hz, 1H), 6.43 – 6.29 (m, 1H), 4.05 (t, J = 6.4 Hz, 2H), 1.97 (t, J = 7.4 Hz,2H), 1.77 – 1.68 (m, 2H), 1.57 – 1.48 (m, 2H), 1.47 – 1.37 (m, 2H), 1.36 –1.27 (m, 2H). ESI(m / z): 497.1808[M+H] + .

[0137] Example 21: 8-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-1H-pyrazole-1-yl)-N-hydroxyoctamide (Id-1), with the following structural formula:

[0138] ;

[0139] Step S1: Preparation of ethyl 8-(4-((tert-Butoxycarbonyl)amino)-1H-pyrazole-1-yl)octanoate: tert-butyl(1H-pyrazole-4-yl)carbamate (0.9 g, 4.9 mmol), cesium carbonate (3.48 g, 9.85 mmol), and ethyl bromooctanoate (1.475 g, 5.88 mmol) were dissolved in acetonitrile (30 mL), and the reaction was carried out at 85 °C for 8 h. The reaction was monitored by TLC until completion. Insoluble matter was removed by diatomaceous earth filtration. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1) to give 1.64 g of yellow solid (95% yield). The structural formula of the prepared ethyl 8-(4-((tert-Butoxycarbonyl)amino)-1H-pyrazole-1-yl)octanoate is as follows:

[0140] ;

[0141] Step S2, preparation of ethyl 8-(4-((3,6-dichloro-1,2,4-triazine-5-yl)amino)-1H-pyrazole-1-yl)octanoate: Dioxane hydrochloride (5 mL) and anhydrous ethanol (2 mL) were added to ethyl 8-(4-((tert-butoxycarbonyl)amino)-1H-pyrazole-1-yl)octanoate (0.24 g, 0.7 mmol), and the reaction was stirred at room temperature for 12 h. After removing the solvent by rotary evaporation, 3,5,6-trichloro-[1,2,4]-thiazide (0.13 g, 0.7 mmol) and triethylamine (0.21 g, 2.1 mmol) were dissolved in tetrahydrofuran (30 mL) and refluxed at 63 °C for 4 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained from the concentration was purified by thin-layer chromatography (petroleum ether: ethyl acetate = 1:2) to give 0.14 g of green solid (yield 50.1%). 1 H NMR (400 MHz, CDCl3) δ(ppm)8.32 (s, 1H), 8.04 (s, 1H), 7.66 (s, 1H), 4.16 – 4.05 (m, 4H), 2.26 (t, J =7.5 Hz, 2H), 1.91 – 1.80 (m, 2H), 1.65 – 1.53 (m, 2H), 1.33 – 1.26 (m, 6H), 1.24 (d, J = 7.0 Hz, 3H). ESI(m / z): 401.1323[M+H] + The structure of the prepared ethyl 8-(4-((3,6-dichloro-1,2,4-triazin-5-yl)amino)-1H-pyrazole-1-yl)octanoate is as follows:

[0142] ;

[0143] Step S3, preparation of ethyl 8-(4-((3-((1H-indole-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-1H-pyrazole-1-yl)octanoate: ethyl 8-(4-((3-((1H-indole-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-1H-pyrazole-1-yl)octanoate (107 mg, 0.27 mmol), 6-aminoindole (35 mg, 0.27 mmol), camphorsulfonic acid (25 mg, 0.11 mmol) and isopropanol (30 mL) were refluxed at 85 °C for 12 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained from the concentration was purified by thin-layer chromatography (dichloromethane:methanol = 100:1) to give 58 mg of a pale yellow solid (yield 43.2%). The structural formula of the prepared ethyl 8-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-1H-pyrazole-1-yl)octanoate is as follows:

[0144] ;

[0145] Step S5, preparation of ((6-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)pyridin-3-yl)oxy)-N-hydroxyheptanamide (Id-1): A methanol solution of potassium hydroxide (8.4 g, 150.1 mmol) was placed in an ice bath, and then a methanol solution of hydroxylamine hydrochloride (7.0 g, 100.6 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 30 min. Insoluble matter was removed by filtration to obtain an NH2OK / methanol solution. Ethyl 8-(4-((3-((1H-indol-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)-1H-pyrazol-1-yl)octanoate (87 mg, 0.17 mmol) was added to 19 mL of the NH2OK / methanol solution, and stirring was continued for 1 h. After the reaction was completed as monitored by TLC, the pH of the reaction mixture was adjusted to 6–7 with acetic acid. The reaction solution was concentrated by vacuum distillation, and the residue was washed with water to give 50 mg of white solid (yield 61.2%). 1H NMR (300MHz, DMSO-d6) δ(ppm) 11.04 (s, 1H), 10.34 (s, 1H), 9.77 (s, 1H), 9.43 (s,1H), 8.68 (s, 1H), 8.07 (s, 1H), 7.81 (s, 1H), 7.77 (s, 1H), 7.49 (d, J = 8.4Hz, 1H), 7.31 – 7.25 (m, 1H), 7.13 (dd, J = 8.5, 1.8 Hz, 1H), 6.45 – 6.32 (m,1H), 3.88 (d, J = 6.8 Hz, 2H), 1.92 (t, J = 7.3 Hz, 2H), 1.68 – 1.52 (m, 2H),1.51 – 1.37 (m, 2H), 1.21 – 1.06 (m, 6H). ESI(m / z): 484.2002[M+H] +

[0146] Example 22: 7-(4-((3-((1H-indol-5-yl)amino)-6-chloro-1,2,4-triazine-5-yl)amino)phenoxy)-N-hydroxyheptanamide (IIb-1), with the following structural formula:

[0147] ;

[0148] The only difference between the synthesis steps S1 to S5 of Example 22 and those of Example 9 is that in step S3, ethyl 2-(4-aminophenoxy)heptanoate is replaced with ethyl 2-(4-aminophenoxy)acetic acid ester, and in step S4, 5-aminoindole is replaced with 6-aminoindole.

[0149] In step S4 of Example 22, ethyl 7-(4-((3-((1H-indole-5-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)heptanoate was prepared by refluxing ethyl 7-(4-((3,6-dichloro-1,2,4-triazin-5-yl)amino)phenoxy)heptanoate (60 mg, 0.146 mmol), 5-aminoindole (19 mg, 0.146 mmol), camphorsulfonic acid (13.5 mg, 0.058 mmol) and isopropanol (30 mL) at 85 °C for 12 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue obtained from the concentration was purified by thin-layer chromatography (dichloromethane:methanol = 100:1) to give 25 mg of a yellow solid (yield 34%). The structural formula of the prepared ethyl 7-(4-((3-((1H-indol-5-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)heptanoate is as follows:

[0150] ;

[0151] Step S5 of Example 22: Preparation of 7-(4-((3-((1H-indole-5-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)-N-hydroxyheptanamide (IIb-6): The only difference from step S5 of Example 9 is that 7-(4-((3-((1H-indole-5-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)heptanyl ester is substituted for ethyl 2-(4-((3-((1H-indole-6-yl)amino)-6-chloro-1,2,4-triazin-5-yl)amino)phenoxy)acetate. 1 H NMR(400 MHz, DMSO-d6) δ(ppm) 10.93 (s, 1H), 10.36 (s, 1H), 9.44 (s, 1H), 9.28(s, 1H), 8.70 (s, 1H), 7.80 (s, 1H), 7.54 (d, J = 8.6 Hz, 2H), 7.28 – 7.25(m, 1H), 7.24 (d, J = 8.6 Hz, 1H), 7.16 (dd, J = 8.7, 2.1 Hz, 1H), 6.93 (d, J= 8.8 Hz, 2H), 6.27 – 6.22 (m, 1H), 3.97 (t, J = 6.4 Hz, 2H), 1.96 (t, J =7.3 Hz, 2H), 1.77 – 1.66 (m, 2H), 1.59 – 1.47 (m, 2H), 1.48 – 1.38 (m, 2H),1.37 – 1.27 (m, 2H). ESI(m / z): 496.1894[M+H] +

[0152] Test example:

[0153] The half-maximal inhibitory concentration (IC50) of the drug on U937 cells was determined using the CCK-8 assay. 50 First, resuscitate U937 cells, quickly thaw them in a 37 °C water bath, then transfer them to centrifuge tubes containing RPMI-1640 complete medium (containing 10% fetal bovine serum and 1% penicillin antibiotics). Centrifuge at 1000 rpm for 5 minutes, discard the supernatant, resuspend the cells, and seed them into culture flasks. Incubate at 37 °C and 5% CO2. After passage to the logarithmic growth phase, collect cells in good condition, count them using a hemocytometer, and adjust the cell concentration to 1 × 10⁻⁶ cells / mL. 5cells / mL, 1×10⁻⁶ per well 4 Cells were seeded in 96-well plates, with a blank control group (culture medium only), a negative control group (cells + culture medium only), and different drug treatment groups, each with 3–5 replicates. Cells were cultured for 1–2 h until stable. Then, serially diluted test drugs were added, with a concentration gradient of 9, typically starting at 100 μM and diluted 1:2. The test drugs refer to the compounds in Table 1 below, including the compounds in the embodiments of this invention, two positive control drugs (SAHA and V11), and their combination. Culture was continued for 48 h. After culture, 10 μL of CCK-8 reagent was added to each well, and incubation was performed for 1–4 h. The absorbance (OD value) was measured at 450 nm using a microplate reader. Cell viability was calculated as follows: viability (%) = (OD of experimental group - OD of blank control group) / (OD of negative control group - OD of blank control group) × 100%. A dose-response curve was plotted with drug concentration on the x-axis and cell viability on the y-axis. Nonlinear regression fitting was performed using GraphPadPrism software to calculate IC50. 50 Values. The experiment was repeated at least 3 times, and the test results are shown in Table 1 below. In the table, A represents a value less than 1 μM, B represents a value greater than 1 μM and less than 5 μM, C represents a value greater than 5 μM and less than 10 μM, and D represents a value greater than 10 μM and less than 20 μM.

[0154] The compounds in the embodiments of the present invention exhibited significant antiproliferative activity against the tumor cell line U-937, and the antiproliferative activity of some compounds was superior to that of the two positive control drugs (SAHA and V11) and their combination.

[0155] Table 1: Inhibitory activity of preferred compounds against U937 cells

[0156] .

Claims

1. A class of small molecules containing indole and triazine groups, characterized in that, The structural formula of the derivative small molecule is as follows: ALB or BLA; Wherein, group A is an indoleamino group or a derivative thereof, and the structure of group A is any one of the structures shown below. "Represents a connection point; ； The L group is a 1,2,4-triazine derivative, covalently bonded to A and B. The structure of the L group can be any one of the structures shown below. "Represents a connection point; ； Wherein, the B group is an N-hydroxyamide derivative with a substituted aromatic group, and the structure of the B group can be any one of the structures shown below. "Represents a connection point; ； Where n is an integer between 1 and 10.

2. The small molecule derivative containing indole and triazine groups as described in claim 1, characterized in that, n=4~9。 3. The small molecule derivative containing an indole group and a triazine group as described in claim 1, characterized in that, The derivative molecule is a class Ia compound, and the structural formula of the class Ia compound is as follows: 。 4. The small molecule derivative containing an indole group and a triazine group as described in claim 1, characterized in that, The derivative small molecule is a class Ib compound, and the structural formula of the class Ib compound is as follows: ； R is selected from any one of -H, -F, -CF3, -OCH3, and -CH3.

5. The small molecule derivative containing an indole group and a triazine group as described in claim 1, characterized in that, The derivative molecule is a class Ic compound, and the structural formula of the class Ic compound is as follows: 。 6. The small molecule derivative containing an indole group and a triazine group as described in claim 1, characterized in that, The derivative small molecule is an Id class compound, and the structural formula of the Id class compound is as follows: 。 7. The small molecule derivative containing an indole group and a triazine group as described in claim 1, characterized in that, The derivative molecule is a class IIa compound, and the structural formula of the class IIa compound is as follows: 。 8. The small molecule derivative containing an indole group and a triazine group as described in claim 1, characterized in that, The derivative molecule is a class IIb compound, and the structural formula of the class IIb compound is as follows: 。 9. The use of the derivative small molecule of any one of claims 1 to 8 or its pharmaceutically acceptable salt, hydrate or prodrug in the preparation of NAE / HDAC dual-target inhibitors.

10. The use of the derivative small molecule of any one of claims 1 to 8 or its pharmaceutically acceptable salt, hydrate or prodrug in the preparation of medicaments for the treatment and / or prevention of cardiovascular diseases, medicaments for the treatment and / or prevention of nervous system diseases, medicaments for the treatment and / or prevention of immune system diseases, medicaments for the treatment and / or prevention of fibrotic diseases, and tumor suppressor drugs.

Images