PROTAC degradation agent and application thereof

Abstract

The invention discloses a PROTAC degradation agent and application thereof, and belongs to the technical field of medicines. The PROTAC degradation agent provided by the invention comprises a compound as shown in a formula (1) or pharmaceutically acceptable salts, solvates, hydrates, prodrugs, stereoisomers, polymorphs, co-crystals and deuterated substances of the compound,

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to PROTAC degraders and applications thereof. Background Art

[0002] Janus kinases (JAKs) are non-receptor tyrosine kinases. The JAK-STAT signaling pathway, formed by Janus kinases and signal transducers and activators of transcription (STATs), influences cell proliferation, differentiation, apoptosis, and immune regulation. JAK and STAT proteins bind to the intracellular portions of type I / II cytokine receptors, which recognize soluble inflammatory mediators such as interleukins and interferons. Upon binding of extracellular ligands to cytokine receptors, JAK proteins are activated and phosphorylate STAT proteins, which then dimerize and translocate to the nucleus, where they activate downstream gene transcription. The four members of the JAK family (JAK1, JAK2, JAK3, and TYK2) and the seven members of the STAT family selectively bind to different cytokine receptors in various combinations. These cytokines include interferon (IFN), interleukin (IL) and hormone signal transduction [such as growth hormone (GH), erythropoietin (EPO), thrombopoietin (TPO), etc.], and cytokines have functions such as regulating autoimmunity, promoting cell growth and blood cell production, and repairing tissue damage.

[0003] Currently, several small-molecule JAK inhibitors are in Phase III clinical trials or have been approved for marketing for the treatment of diseases such as rheumatoid arthritis (RA), psoriasis (PS), and atopic dermatitis. JAK inhibitors can be divided into first-generation non-selective inhibitors and second-generation selective inhibitors. The former target two or more of the four JAK isoforms—JAK1, JAK2, JAK3, and TYK2. However, traditional JAK inhibitors currently have several major limitations, primarily manifested in the following aspects: 1. Insufficient targeting: Traditional JAK inhibitors typically inhibit multiple JAK family members, resulting in off-target effects and increased side effects; 2. Drug resistance: Long-term use of JAK inhibitors may lead to target mutations, resulting in drug resistance; 3. Low bioavailability: Some JAK inhibitors have low bioavailability, requiring frequent dosing, which affects patient compliance.

[0004] Proteolysis targeting chimera (PROTAC) is a kind of bifunctional molecule with two recruiting ligands combined together by a linker. One ligand can recruit E3 ubiquitin ligase, and the other ligand can specifically recruit target protein of interest (POI). When the ternary complex is formed, PROTAC can recruit E3 ligase to the target protein of interest, which is spatially conducive to substrate polyubiquitination, thereby leading to the subsequent degradation of POI. Compared with occupancy-based pharmacology, PROTAC technology has several advantages. First, due to the catalytic nature of PROTAC, PROTAC can achieve a therapeutic effect comparable to occupancy-based inhibitors at a lower concentration. Second, PROTAC can be used as a new method to target “undruggable” proteins, such as transcription factors, scaffold proteins and non-enzyme proteins. Third, PROTAC can selectively bind to targets that are difficult to achieve with some small molecules. Therefore, how to develop new JAK degraders using PROTAC technology to achieve more precise and effective treatment has become a technical problem to be solved in the field. SUMMARY

[0005] To solve the above technical problems, the present application provides a PROTAC degrader and its application.

[0006] To achieve the above purpose, the present application provides the following technical solutions.

[0007] The present application provides a PROTAC degrader, comprising a compound represented by formula (1) or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, deuteride thereof,

[0008] Formula (1);

[0009] Wherein, L is , or ; R is H, , or ; G1 and G2 are independently a bond, CH2 or O, and G1 and G2 are not a bond at the same time; n and m are independently 1, 2, 3 or 4.

[0010] Further, the L is , , , , , , , or .

[0011] Further, the structure formula of the PROTAC degrader is 、 、 or .

[0012] Further, the structural formula of the PROTAC degrader is:

[0013]

[0014]

[0015]

[0016]

[0017]

[0018] .

[0019] The application provides a preparation method of the PROTAC degrader.

[0020] (1) Preparation of the compound of formula VI:

[0021] ;

[0022] a) mixing the compound of formula I with N,N-dimethylformamide dimethyl acetal and then reacting at 50-120 DEG C. for 10-14 h. to obtain the compound of formula II;

[0023] b) mixing the compound of formula III with cyanamide and hydrochloric acid and then reacting at 50-120 DEG C. for 8-12 h. to obtain the compound of formula IV;

[0024] c) mixing the compound of formula II obtained in step a) with the compound of formula IV obtained in step b) and then reacting at 80-120 DEG C. for 40-60 h. to obtain the compound of formula V;

[0025] d) hydrolyzing the compound of formula V obtained in step c) at 20-80 DEG C. for 1-8 h. to obtain the compound of formula VI;

[0026] (2) Preparation of the compound of formula XI:

[0027] ;

[0028] e) reacting the compound of formula VII and the compound of formula VIII in the presence of N,N-diisopropylethylamine at 50-100° C. for 2-8 hours to obtain a compound of formula IX;

[0029] f) reacting the compound of formula IX obtained in step e) with an HCl-dioxane solution at 20-50° C. for 1-3 h to obtain a compound of formula X;

[0030] g) reacting the compound of formula X obtained in step f) with the compound of formula VI obtained in step d) at 20-50° C. for 8-12 h to obtain a compound of formula XI;

[0031] Or (3) Preparation of the compound of formula XV:

[0032] ;

[0033] h) mixing the compound of formula X obtained in step f) with the compound of formula XII and reacting the mixture at 20-50° C. for 8-12 hours to obtain a compound of formula XIII;

[0034] i) mixing the compound of formula XIII obtained in step h) with an HCl-dioxane solution and reacting the mixture at 20-50° C. for 1-3 hours to obtain a compound of formula XIV;

[0035] j) mixing the compound of formula XIV obtained in step i) with the compound of formula VI obtained in step d) and reacting the mixture at 20-50° C. for 8-12 hours to obtain a compound of formula XV;

[0036] Or (4) Preparation of the compound of formula XXI:

[0037] ;

[0038] k) mixing the compound of formula VI obtained in step d) with the compound of formula XVI and reacting them at 20-50° C. for 10-14 hours to obtain a compound of formula XVII;

[0039] i) mixing the compound of formula XVII obtained in step k) with an HCl-dioxane solution and reacting the mixture at 20-50° C. for 1-3 hours to obtain a compound of formula XVIII;

[0040] m) reacting the compound of formula XIX in the presence of a Dess-Martin periodinane at 20-50° C. for 2-4 h to obtain a compound of formula XX;

[0041] n) reacting the compound of formula XX obtained in step m) and the compound of formula XVIII obtained in step i) with glacial acetic acid and sodium triacetoxyborohydride at 20-50° C. for 14-18 hours to obtain a compound of formula XXI;

[0042] In formula VIII, formula IX, formula X, formula XI, formula XIII, formula XIV, formula XV, formula XIX, formula XX and formula XXI, R is H, , or ; G1 and G2 are independently a bond, CH2 or O, and G1 and G2 are not a bond at the same time; n and m are independently 1, 2, 3 or 4.

[0043] The application further provides a pharmaceutical composition comprising the PROTAC degrader described in the above technical solution and a pharmaceutically acceptable auxiliary or excipient.

[0044] The application further provides use of the PROTAC degrader or the pharmaceutical composition described in the above technical solution in preparation of a drug for preventing and / or treating an inflammatory disease, a respiratory disease, a skin disease, an immune system disease or colitis.

[0045] Further, the inflammatory disease is an inflammatory skin disease; the respiratory disease is chronic obstructive pulmonary disease, colitis or asthma; the skin disease is psoriasis or specific dermatitis; the immune system disease is systemic lupus erythematosus or rheumatoid arthritis.

[0046] The application further provides use of the PROTAC degrader or the pharmaceutical composition described in the above technical solution in preparation of a drug for preventing and / or treating a JAK-mediated disease or a JAK inhibitor.

[0047] Further, the JAK is JAK1 or JAK2.

[0048] Compared with the prior art, the application has the following advantages and technical effects:

[0049] The PROTAC degrader provided by the application has anti-inflammatory activity, can effectively relieve inflammatory reactions, has strong selectivity for JAK1 / 2 subtypes and can dose-dependently degrade JAK1 proteins. BRIEF DESCRIPTION OF DRAWINGS

[0050] The accompanying drawings, which form a part of the present application, are included to provide a further understanding of the application, and are incorporated herein for explanation by reference. These drawings illustrate exemplary embodiments of the present application and, together with the description, serve to explain the present application:

[0051] Figure 1 are the screening results of JAK1 protein degradation activity of each compound (A) and the heat map of JAK1 protein degradation rate (B);

[0052] Figure 2 are the results of cytotoxicity experiments of compounds A8, C2, C4 and C6;

[0053] Figure 3 Screening results (left) and degradation rate curves (right) of JAK1 protein degradation activity in RAW264.7 cells after treatment with compounds A8 (A), C2 (B), and C4 (C);

[0054] Figure 4 The anti-inflammatory effects of compounds A8, C2, and C4 in LPS-induced RAW264.7 cells, where A represents the NO production, B represents the IL-6 level, and C represents the TNF-α level;

[0055] Figure 5 The screening results (left) and degradation rate curve (right) of compound A8 on the degradation activity of JAK2 (A), JAK3 (B) and TYK2 (C);

[0056] Figure 6 The results show the improvement effect of compound A8 on the symptoms of DSS-induced colitis in mice, where A is a schematic diagram of the animal experimental design, B is the weight change during the experiment, C is the disease activity index (DAI) score, D is the colon length, E is the spleen index, F is a representative picture of the colon in each group, and G is a representative picture of the spleen in each group. The values ​​are expressed as mean ± standard deviation (n=8). Compared with the model group, *P<0.05, **P<0.01, ***P<0.001;

[0057] Figure 7 The intestinal histopathological evaluation results of DSS-induced colitis mice, where A is the H&E staining and Alcian blue-PAS staining image of the colon, and B is the immunohistochemical staining result of the colon tight junction protein ZO-1 and Occludin;

[0058] Figure 8 Figure 3: Inflammatory factor levels, blood routine examination, and immunohistochemical analysis results of mice with DSS-induced colitis. A shows the levels of IL-6, IL-10, IL-1β, and TNF-α in serum (ELISA assay); B shows the levels of IL-6, IL-10, IL-1β, and TNF-α in colon tissue (ELISA assay); C shows the subtype analysis of peripheral blood immune cells, including total white blood cells (WBCs), lymphocytes, monocytes, and neutrophils. The dotted lines indicate the upper and lower limits of the normal range; D shows the immunohistochemical staining of MPO and iNOS in colon tissue. Positive signals appear brown. Scale bar = 200 μm. Values ​​are expressed as mean ± standard deviation (n = 8). Compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001. DETAILED DESCRIPTION

[0059] With reference to the accompanying drawings on the basis of the embodiments of the present application, the technical solutions in the embodiments of the present application will be described clearly and completely. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments of the present application, all the other embodiments obtained by a person of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0060] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the present application will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0061] Unless otherwise specified, the raw materials in the embodiments of the present application are obtained by commercial channels.

[0062] Synthesis of intermediates 3a-3f in Example 1

[0063] The reaction route is: ;

[0064] The preparation process is:

[0065] (1) Synthesis of compounds 2a-2f

[0066] The reaction route is: ;

[0067] Dissolve 2.0 g of compound 1 (7.24 mmol, 1.0 equiv) in 10.0 mL of N,N-dimethylformamide solution, then add the amine reagent (8.69 mmol, 1.2 equiv) and N,N-diisopropylethylamine (DIPEA, 14.48 mmol, 2.0 equiv) into the above solution, heat the reaction at 90 ℃ for 4 h, and monitor the reaction by thin layer chromatography. After the reaction is completed, quench with water, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, concentrate the organic phase, and purify by flash column chromatography to obtain compounds 2a-2f;

[0068] The structural formula of compounds 2a-2f is:

[0069]

[0070] ;

[0071] Synthesis of compound 2a: the amine reagent is selected as single Boc-ethylenediamine (CAS No.: 57260-73-8), and compound 2a is obtained, with a yield of 25%; 1H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 7.57 (dd, J = 8.5,7.0 Hz, 1H), 7.14 (d, J = 8.5 Hz, 1H), 7.05–6.95 (m, 2H), 6.71 (t, J = 6.0Hz, 1H), 5.05 (dd, J = 12.5, 5.5 Hz, 1H), 3.39–3.34 (m, 2H), 3.14–3.09 (m,2H), 2.92–2.85 (m, 1H), 2.63–2.51 (m, 2H), 2.04–1.99 (m, 1H), 1.36 (s, 9H).ESI-MS m / z: The calculated value is C 20 H 24 O6N4Na + [M + Na] + , 439.2; the measured value is 439.2.

[0072] Synthesis of compound 2b: N-Boc-1,3-propylenediamine (CAS No. 75178-96-0) was selected as an amine reagent to obtain compound 2b in a yield of 25%; 1 H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 7.57 (dd, J =8.5, 7.0 Hz, 1H), 7.08 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 7.0 Hz, 1H), 6.91(t, J = 6.0 Hz, 1H), 6.66 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.5, 5.5 Hz,1H), 3.33–3.29 (m, 2H), 3.02–2.97 (m, 2H), 2.92–2.85 (m, 1H), 2.63–2.51 (m,2H), 2.05–2.00 (m, 1H), 1.69–1.63 (m, 2H), 1.37 (s, 9H). ESI-HRMS m / z: calcd for C 21 H 26 O6N4Na + [M + Na] + , 453.1745; the measured value is 453.1731.

[0073] Synthesis of compound 2c: tert-butyl (4-aminobutyl)carbamate (CAS No. 68076-36-8) was selected as an amine reagent to obtain compound 2c in a yield of 27%; 1 H NMR (500 MHz, DMSO-d6) δ 11.08 (s, 1H), 7.57(dd, J = 8.5, 7.0 Hz, 1H), 7.10 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 7.0 Hz,1H), 6.82 (t, J = 6.0 Hz, 1H), 6.54 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 13.0,5.5 Hz, 1H), 3.31–3.26 (m, 2H), 2.98–2.92 (m, 2H), 2.91–2.84 (m, 1H), 2.62–2.51 (m, 2H), 2.05–2.00 (m, 1H), 1.57–1.51 (m, 2H), 1.48–1.41 (m, 2H), 1.36(s, 9H). ESI-HRMS m / z: calculated for C 22 H 28 O6N4Na + [M + Na] + , 467.1901; the measured value is 467.1876.

[0074] Synthesis of compound 2d: tert-butyl N-(5-aminopentyl)carbamate (CAS No. 51644-96-3) was selected as an amine reagent to obtain compound 2d in a yield of 42%; 1H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 7.57(dd, J = 8.5, 7.0 Hz, 1H), 7.08 (d, J = 8.5 Hz, 1H), 7.01 (d, J = 7.0 Hz,1H), 6.77 (t, J = 6.0 Hz, 1H), 6.51 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 13.0,5.5 Hz, 1H), 3.29–3.25 (m, 2H), 2.94–2.90 (m, 2H), 2.89–2.84 (m, 1H), 2.62–2.51 (m, 2H), 2.06–2.00 (m, 1H), 1.59–1.53 (m, 2H), 1.43–1.38 (m, 2H), 1.36(s, 9H), 1.33–1.28 (m, 2H). ESI-HRMS m / z: calculated for C 23 H 30 O6N4Na + [M + Na] + ,481.2058; the measured value is 481.2039.

[0075] Synthesis of compound 2e: amine reagent selected (6-aminohexyl) carbamic acid tert-butyl ester (CAS No.: 51857-17-1) to obtain compound 2e in a yield of 47%; 1 H NMR (500 MHz, DMSO-d6) δ 11.08 (s, 1H), 7.57(dd, J = 8.5, 7.0 Hz, 1H), 7.08 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 7.0 Hz,1H), 6.75 (t, J = 6.0 Hz, 1H), 6.52 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 13.0,5.5 Hz, 1H), 3.30–3.26 (m, 2H), 2.92–2.88 (m, 2H), 2.88–2.84 (m, 1H), 2.61–2.51 (m, 2H), 2.05–2.00 (m, 1H), 1.59–1.52 (m, 2H), 1.40–1.37 (m, 2H), 1.36(s, 9H), 1.34–1.26 (m, 4H). ESI-HRMS m / z: calculated for C 24 H32 O6N4Na + [M + Na] + , 527.2112; found 527.2097.

[0076] The synthesis of compound 2f: tert-butyl 2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate (CAS No.: 153086-78-3) was selected as the amine reagent to give compound 2f in 41% yield. 1 H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 7.58 (dd, J = 8.5, 7.0 Hz, 1H), 7.13 (d, J = 8.5 Hz, 1H), 7.03 (d, J = 7.0 Hz, 1H), 6.72 (t, J = 6.0 Hz, 1H), 6.60 (t, J = 6.0 Hz, 1H), 5.06 (dd, J = 13.0, 5.5 Hz, 1H), 3.62 (t, J = 5.5 Hz, 2H), 3.57–3.54 (m, 2H), 3.52–3.50 (m, 2H), 3.48–3.45 (m, 2H), 3.38 (t, J = 6.0 Hz, 2H), 3.08–3.03 (m, 2H), 2.92–2.84 (m, 1H), 2.62–2.51 (m, 2H), 2.05–2.00 (m, 1H), 1.35 (s, 9H). ESI-HRMS m / z: calcd for C 24 H 32 O8N4Na + [M + Na] + , 527.2112; found 527.2097.

[0077] (2) Synthesis of compounds 3a-3f

[0078] The reaction scheme is: ;

[0079] The 500 mg of compound 2a-2f obtained in step (1) was dissolved in 9.0 mL of DCM, 3 mL of hydrogen chloride-1,4 dioxane solution was added, and the reaction was carried out at room temperature for 2 h. After the reaction was completed, the solid was collected by filtration and dried to obtain yellow-green solid 3a-3f in 75-89% yield.

[0080] Example 2 Synthesis of compound 5a

[0081] The reaction scheme is:

[0082] ;

[0083] The preparation process is:

[0084] (1) Synthesis of compound 4a

[0085] The reaction scheme is: ;

[0086] 500 mg of compound 3f prepared in Example 1 (1.1 mmol, 1.0 equiv) and 2-(4-(tert-butoxycarbonyl)piperazin-1-yl)acetic acid (CAS No.: 156478-71-6) (1.21 mmol, 1.1 equiv) were dissolved in 5.0 mL of N,N-dimethylformamide, and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 1.1 mmol, 1.0 equiv, CAS No: 148893-10-1) and N,N-diisopropylethylamine (DIPEA, 5.5 mmol, 5.0 equiv) were added. The reaction was stirred at room temperature for 10 h, and thin layer chromatography was used for monitoring. After the reaction was completed, water was added for quenching, and the organic phase was dried with anhydrous sodium sulfate, the organic phase was concentrated, and yellow-green solid compound 4a was obtained by flash column chromatography, with a yield of 44%.

[0087] The obtained compound 4a was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were as follows: 1H NMR (500MHz, DMSO-d6) δ 11.08 (s, 1H), 7.71 (t, J = 6.0 Hz, 1H), 7.58 (dd, J = 8.5, 7.0 Hz, 1H), 7.14 (d, J = 8.5 Hz, 1H), 7.04 (d, J = 7.0 Hz, 1H), 6.61 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 13.0, 5.5 Hz, 1H), 3.62 (t, J = 5.5 Hz, 2H), 3.57–3.51 (m, 4H), 3.49–3.45 (m, 2H), 3.43 (t, J = 6.0 Hz, 2H), 3.32–3.29 (m, 4H), 3.26–3.22 (m, 2H), 2.91 (s, 2H), 2.89–2.84 (m, 1H), 2.62–2.51 (m, 2H), 2.35 (t, J = 5.5 Hz, 4H), 2.05–2.00 (m, 1H), 1.37 (s, 9H). ESI-HRMS m / z: Calcd for C 30 H 43 O9N6 + [M + H] + , 631.3086; Found, 631.3058.

[0088] (2) Synthesis of compound 5a

[0089] The reaction scheme is as follows: ;

[0090] 500 mg of compound 4a prepared in step (1) was dissolved in 9.0 mL of DCM, and 3 mL of hydrogen chloride-1,4 dioxane solution was added to the above solution, and the reaction was carried out at room temperature for 2 h. After the reaction was completed, the solid was collected by filtration and dried to obtain a yellow-green solid 5a, yield: 78 %.

[0091] The obtained compound 5a was identified by nuclear magnetic resonance spectrum and mass spectrometry, and the identification result was as follows: 1H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.89 (br, 1H), 8.61 (br, 1H), 7.59 (dd, J = 8.5, 7.0 Hz, 1H), 7.15 (d, J = 8.5 Hz, 1H), 7.04 (d, J = 7.0 Hz, 1H), 6.60 (br, 1H), 5.05 (dd, J = 13.0, 5.5 Hz, 1H), 3.91-3.80 (m, 2H), 3.63-3.61 (m, 2H), 3.58-3.53 (m, 6H), 3.49-3.44 (m, 6H), 3.39-3.34 (m, 4H), 3.29-3.26 (m, 2H), 2.93-2.85 (m, 1H), 2.63-2.51 (m, 2H), 2.05-2.00 (m, 1H). ESI-HRMS m / z: calcd for C 25 H 35 O7N6 + [M + H] + , 531.2562; found 531.2537.

[0092] Example 3. Synthesis of compounds 8a-8b

[0093] The reaction scheme is: ;

[0094] The preparation process is:

[0095] (1) Synthesis of compounds 7a-7b

[0096] The reaction scheme is: ;

[0097] Dissolve 1.0 g of compound 6 (3.65 mmol, 1.0 equiv) in 5.0 mL of N,N- dimethylformamide, add 5-bromo-1-pentanol or 6-bromo-1-hexanol (7.30 mmol, 2.0 equiv), then add NaHCO3(14.60 mmol, 4.0 equiv) and KI (0.37 mmol, 0.1 equiv), and react at 80 °C for 8 h, monitoring by thin layer chromatography. After the reaction is completed, quench with water, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, concentrate the organic phase, and purify by flash column chromatography to obtain white solid compounds 7a-7b.

[0098] The structural formula of compounds 7a-7b is: ;

[0099] Synthesis of compound 7a: 5-bromo-1-pentanol was selected as the reactant to obtain compound 7a in a yield of 48%; 1 H NMR (500MHz, DMSO-d6) δ 11.11 (br, 1H), 7.80 (dd, J = 8.5, 7.0 Hz, 1H), 7.50 (d, J =8.5 Hz, 1H), 7.43 (d, J = 7.5 Hz, 1H), 5.08 (dd, J = 13.0, 5.5 Hz, 1H), 4.40(br, 1H), 4.19 (t, J = 6.5 Hz, 2H), 3.42–3.40 (m, 2H), 2.92–2.84 (m, 1H), 2.63–2.50 (m, 2H), 2.06–2.00 (m, 1H), 1.79–1.73 (m, 2H), 1.51–1.45 (m, 4H).ESI-MS m / z: calculated for C 18 H 21 O6N2 + [M + H] + , 361.1; the measured value is 361.1.

[0100] Synthesis of compound 7b: 6-bromo-1-hexanol was selected as the reactant to obtain compound 7b in a yield of 45%; 1 H NMR (500MHz, DMSO-d6) δ 11.10 (s, 1H), 7.80 (dd, J = 8.5, 7.0 Hz, 1H), 7.50 (d, J =8.5 Hz, 1H), 7.43 (d, J = 7.0 Hz, 1H), 5.08 (dd, J = 13.0, 5.5 Hz, 1H), 4.35(t, J = 5.5 Hz, 1H), 4.19 (t, J = 6.5 Hz, 2H), 3.42–3.38 (m, 2H), 2.92–2.84(m, 1H), 2.63–2.50 (m, 2H), 2.06–2.00 (m, 1H), 1.78–1.72 (m, 2H), 1.48–1.41 (m, 4H), 1.38–1.33 (m, 2H). ESI-HRMS m / z: calculated for C 19 H 22 O6N2Na + [M + Na] +397.1370; found 397.1350.

[0101] (2) Synthesis of compounds 8a-8b

[0102] The reaction scheme is as follows: ;

[0103] The compound 7a-7b prepared in step (1) (2.0 mmol, 1.0 equiv) was dissolved in dichloromethane, Dess-Martin Periodinane (CAS No: 87413-09-0, 4.0 mmol, 2.0 equiv) was added, stirred at room temperature for 3 h, then saturated sodium bicarbonate and saturated sodium thiosulfate solution were added, stirred for 10 min, the organic phase was collected, the aqueous phase was extracted with dichloromethane, the organic phase was dried with anhydrous sodium sulfate, the organic phase was concentrated, and white solid compound 8a-8b was obtained by flash column chromatography.

[0104] The structural formula of compound 8a, 8b is as follows: ;

[0105] Synthesis of compound 8a: reactant selection 7a, to obtain compound 8a, yield 53%; 1 H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.69 (t, J = 1.6 Hz, 1H), 7.81 (dd, J = 8.4, 7.2 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.44 (d, J = 7.2 Hz, 1H), 5.08 (dd, J = 12.8, 5.4 Hz, 1H), 4.21 (t, J = 6.0 Hz, 2H), 2.93-2.83 (m, 1H), 2.63-2.50 (m, 4H), 2.07-1.97 (m, 1H), 1.85-1.65 (m, 4H). ESI-HRMS m / z: calculated for C 18 H 19 O6N2 + [M+ H] + , 359.1238; found 359.1238.

[0106] Synthesis of compound 8b: reactant selection 7b, to obtain compound 8b, yield 55%; 1H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.67 (t, J = 1.6 Hz, 1H), 7.80 (dd, J = 8.4, 7.2 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.44 (d, J = 7.2 Hz, 1H), 5.08 (dd, J = 12.8, 5.4 Hz, 1H), 4.19 (t, J = 6.4 Hz, 2H), 2.93 - 2.84 (m, 1H), 2.62 - 2.51 (m, 2H), 2.46 (td, J = 7.2, 1.6 Hz, 2H), 2.06 - 1.98 (m, 1H), 1.80 - 1.72 (m, 2H), 1.65 - 1.56 (m, 2H), 1.49 - 1.42 (2H). ESI-HRMS m / z: calcd for C 19 H 21 O6N2 + [M + H] + ,373.1394; Found, 373.1395.

[0107] Example 4 Synthesis of compounds 14a-14d

[0108] The reaction scheme is: ;

[0109] The preparation process is:

[0110] (1) Synthesis of compound 10

[0111] The reaction scheme is: ;

[0112] Compound 9 (4.0 g, 22.4 mmol, 1.0 equiv) was dissolved in 20 mL of N,N- dimethylformamide dimethyl acetal (DMF-DMA) and heated to reflux at 85 °C for 12 h. After the reaction was completed, filtration was performed to obtain 4.5 g of yellow solid compound 10, yield: 86%.

[0113] The obtained compound 10 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1H NMR (500MHz, DMSO-d6) δ 6.85 (d, J = 9.0 Hz, 2H), 6.79 (d, J = 8.0 Hz, 2H), 6.58–5.49 (m, 4H), 3.72 (t, J = 4.5 Hz, 4H), 3.01 (t, J = 4.5 Hz, 4H). ESI-HRMS m / z: calcd for C 13 H 16 O3N + [M + H] + , 234.1125; found 234.1124.

[0114] (2) Synthesis of compounds 12a-12d

[0115] The reaction scheme is: ;

[0116] The amine reagent compounds 11a-11d (20.0 mmol, 1.0 equiv) and cyanamide (CAS No.: 420-04-2, 60.0 mmol, 3.0 equiv) were dissolved in 30 mL of ethanol, 8 mL of concentrated hydrochloric acid was added and heated to reflux at 85 °C for 10 h. After the reaction was completed, the ethanol was removed by concentration, the pH was adjusted to 12 by adding aqueous sodium carbonate solution, and the crude product was obtained by concentration. The intermediate 12a-12d was obtained by flash column chromatography.

[0117] The structural formula of compounds 12a-12d is:

[0118] ;

[0119] Synthesis of compound 12a: reactant selected 11a, to obtain compound 12a, yield: 79%; 1 H NMR (500MHz, DMSO-d6) δ 6.85 (d, J = 9.0 Hz, 2H), 6.79 (d, J = 8.0 Hz, 2H), 6.58–5.49 (m, 4H), 3.72 (t, J = 4.5 Hz, 4H), 3.01 (t, J = 4.5 Hz, 4H). ESI-HRMS m / z: calcd for C 11 H 17 ON4 + [M + H] + , 221.1397; found 221.1399.

[0120] Synthesis of compound 12b: reactant selected 11b, to obtain compound 12b, yield: 67%;1 H NMR (500MHz, Methanol-d4) δ 7.50–7.45 (m, 2H), 7.39–7.34 (m, 1H), 7.31–7.27 (m, 2H),5.71 (s, 1H), NH (3H, not observed). ESI-HRMS m / z: Calcd for C7H 10 N3 + [M + H] + , 136.0869; Found, 136.0870.

[0121] Synthesis of compound 12c: Reactant selection 11c afforded compound 12c in yield: 68%; 1 H NMR (500MHz, Methanol-d4) δ 7.75–7.71 (m, 2H), 7.66–7.61 (m, 2H), 7.48–7.43 (m, 2H),7.39–7.33 (m, 3H), NH (4H, not observed). ESI-HRMS m / z: Calcd for C 13 H 14 N3 + [M + H] + , 212.1182; Found, 212.1184.

[0122] Synthesis of compound 12d: Reactant selection 11d afforded compound 12d in yield: 86%; 1 H NMR (500MHz, DMSO-d6) δ 7.14 (d, J = 8.5 Hz, 2H), 6.92 (d, J = 8.0 Hz, 2H), 2.48–2.40 (m, 1H), 2.10–1.85 (m, 1H), 1.82–1.74 (m, 4H), 1.72–1.67 (m, 1H), 1.40–1.32 (m, 4H), NH (4H, not observed). ESI-HRMS m / z: Calcd for C 13 H 20 N3 + [M + H] + , 218.1652; Found, 218.1654.

[0123] (3) Synthesis of compounds 13a-13d

[0124] The reaction scheme is: ;

[0125] To a solution of compound 10 (15.0 mmol, 1.0 equiv) prepared in step (1) and compound 12a-12d (15.0 mmol, 1.0 equiv) prepared in step (2) in 40 mL of acetonitrile, heat to reflux at 100 °C for 48 h, TLC test. When the reaction is completed, cool the reaction mixture to room temperature, solid is formed, filter to get the crude product, and finally purify by flash column chromatography to get intermediates 13a-13d.

[0126] The structural formula of compounds 13a-13d is: ;

[0127] Synthesis of compound 13a: reactant selection 12a, to get compound 13a, yield: 74%; 1 H NMR (400MHz, DMSO-d6) δ 9.55 (s, 1H), 8.55 (d, J = 5.2 Hz, 1H), 8.30–8.26 (m, 2H),8.13–8.08 (m, 2H), 7.69–7.64 (m, 2H), 7.40 (d, J = 5.2 Hz, 1H), 6.96–6.91 (m,2H), 3.89 (s, 3H), 3.77–3.72 (m, 4H), 3.07–3.02 (m, 4H). ESI-HRMS m / z: calculated for C 22 H 23 O3N4 + [M + H] + , 391.1765; found 391.1762.

[0128] Synthesis of compound 13b: reactant selection 12b, to get compound 13b, yield: 22%; 1 H NMR (500MHz, DMSO-d6) δ 9.76 (s, 1H), 8.61 (d, J = 5.0Hz, 1H), 8.32–8.27 (m, 2H),8.15–8.09 (m, 2H), 7.83 (d, J = 7.5Hz, 2H), 7.48 (d, J = 5.0Hz, 1H), 7.35–7.29 (m, 2H), 6.98 (t, J = 7.5Hz, 1H), 3.90 (s, 3H). ESI-HRMS m / z: calculated for C 18 H 16 O2N3 +[M + H] + Found 306.1238.

[0129] Synthesis of compound 13c: Reactant selection 12c afforded compound 13c in 57% yield; 1 H NMR (500MHz, DMSO-d6) δ 9.68 (s, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.31- 8.25 (m, 2H), 8.13-8.06 (m, 2H), 7.73-7.67 (m, 2H), 7.43 (d, J = 5.0 Hz, 1H), 7.15 (d, J = 8.5 Hz, 2H), 3.89 (s, 3H), 2.47-2.38 (m, 1H), 1.78 (d, J = 9.5 Hz, 4H), 1.72-1.66 (m, 1H), 1.41-1.32 (m, 4H), 1.26-1.21 (m, 1H). ESI-HRMS m / z: Calcd for C 24 H 19 O2N3Na + [M + Na] + Found 404.1358.

[0130] Synthesis of compound 13d: Reactant selection 12d afforded compound 13d in 76% yield; 1 H NMR (500MHz, DMSO-d6) δ 9.68 (s, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.31- 8.25 (m, 2H), 8.13-8.06 (m, 2H), 7.73-7.67 (m, 2H), 7.43 (d, J = 5.0 Hz, 1H), 7.15 (d, J = 8.5 Hz, 2H), 3.89 (s, 3H), 2.47-2.38 (m, 1H), 1.78 (d, J = 9.5 Hz, 4H), 1.72-1.66 (m, 1H), 1.41-1.32 (m, 4H), 1.26-1.21 (m, 1H). ESI-HRMS m / z: Calcd for C 24 H 25 O2N3Na + [M + Na] + Found 410.1857.

[0131] (4) Synthesis of compounds 14a-14d

[0132] The reaction scheme is: ;

[0133] The compound 13a-13d (4.0 mmol, 1.0 equiv) prepared in step (3) was weighed into 20 mL of a mixed solvent of ethanol and water in a volume ratio of 1:1, 1.6 g of sodium hydroxide (40.0 mmol, 10.0 equiv) was added, and the reaction was stirred at 50 °C for 4 h, which was monitored by TLC. After the reaction was completed, the ethanol was removed by rotary evaporation, and the pH was adjusted to 6 with dilute hydrochloric acid (concentration of 1 mol / L). Filtration under suction gave a solid which was dried to obtain a yellow solid compound 14a-14d.

[0134] The structural formula of compound 14a-14d is:

[0135] ;

[0136] Synthesis of compound 14a: the reactant selected was 13a to obtain compound 14a, yield: 91%; 1 H NMR (500MHz, DMSO-d6) δ 13.17 (br, 1H), 9.60 (s, 1H), 8.55 (d, J = 5.0 Hz, 1H), 8.26(d, J = 8.0 Hz, 2H), 8.09 (d, J = 8.0 Hz, 2H), 7.71 (d, J = 8.5 Hz, 2H), 7.40(d, J = 5.0 Hz, 1H), 7.05 (s, 2H), 3.79 (t, J = 4.5 Hz, 4H), 3.13 (s, 4H).ESI-HRMS m / z: calculated C 21 H 21 O3N4 + [M + H] + , 377.1608; found 377.1605.

[0137] Synthesis of compound 14b: the reactant selected was 13b to obtain compound 14b, yield: 79%; 1H NMR (500MHz, DMSO-d6) δ 13.18 (br, 1H), 9.75 (s, 1H), 8.60 (d, J = 5.0 Hz, 1H), 8.29–8.25 (m, 2H), 8.12–8.08 (m, 2H), 7.86–7.81 (m, 2H), 7.46 (d, J = 5.0 Hz, 1H),7.36–7.29 (m, 2H), 7.00–6.95 (m, 1H). ESI-HRMS m / z: Calcd for C 17 H 14 O2N3 + [M + H] + , 292.1081; Found, 292.1077.

[0138] Synthesis of compound 14c: Reactant selection 13c afforded compound 14c in yield: 80%; 1 H NMR (500MHz, DMSO-d6) δ 9.83 (s, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.16–8.11 (m, 2H),8.07–8.02 (m, 2H), 8.00–7.94 (m, 2H), 7.66 (td, J = 7.5, 7.0, 2.0 Hz, 4H),7.46–7.41 (m, 3H), 7.33–7.28 (m, 1H), NH (1H, not observed). ESI-HRMS m / z: Calcd for C 23 H 18 O2N3 + [M + H] + , 368.1394; Found, 368.1392.

[0139] Synthesis of compound 14d: Reactant selection 13d afforded compound 14d in yield: 65%; 1H NMR (500MHz, DMSO-d6) δ 13.16 (br, 1H), 9.64 (s, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.29–8.24 (m, 2H), 8.12–8.07 (m, 2H), 7.74–7.69 (m, 2H), 7.43 (d, J = 5.0 Hz, 1H),7.18–7.14 (m, 2H), 2.48–2.40 (m, 1H), 1.82–1.75 (m, 4H), 1.72–1.66 (m, 1H),1.43–1.32 (m, 4H), 1.26–1.21 (m, 1H). ESI-HRMS m / z: Calcd for C 23 H 24 O2N3 + [M + H] + ,374.1863; Found, 374.1859.

[0140] Example 5. Synthesis of compounds 16a-16d

[0141] The reaction scheme is:

[0142] ;

[0143] The preparation process is:

[0144] (1) Synthesis of compounds 15a-15d

[0145] The reaction scheme is:

[0146] ;

[0147] The compound 14a-14d (1.5 mmol, 1.0 equiv) prepared in Example 4 and piperazine-1-carboxylic acid tert-butyl ester (CAS No.: 57260-71-6, 1.65 mmol, 1.1 equiv) were dissolved in DCM, and N-hydroxy-7-azabenzotriazole (CAS No: 39968-33-7, HOAT, 1.65 mmol, 1.1 equiv), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI, 1.65 mmol, 1.1 equiv) and N-methylmorpholine (NMM, 1.65 mmol, 1.1 equiv) were added at room temperature, and the mixture was stirred at room temperature for 12 h. After the reaction was completed, the crude product was concentrated under vacuum, and then yellow solid intermediates 15a-15d were obtained by flash column chromatography;

[0148] The structural formulas of compounds 15a-15d are:

[0149] ;

[0150] Synthesis of compound 15a: reactant 14a was selected to give compound 15a, yield: 71%; 1 H NMR (500 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.52 (d, J = 5.0 Hz, 1H), 8.23–8.18 (m, 2H),7.69–7.64 (m, 2H), 7.59–7.55 (m, 2H), 7.36 (d, J = 5.0 Hz, 1H), 6.95–6.91 (m,2H), 3.76–3.72 (m, 4H), 3.68–3.52 (m, 2H), 3.49–3.26 (m, 6H), 3.06–3.02 (m,4H), 1.41 (s, 9H). ESI-HRMS m / z: calcd for C 30 H 37 O4 N6 + [M + H] + , 545.2871; the measured value is 545.2872.

[0151] Synthesis of compound 15b: reactant 14b was selected to give compound 15b, yield: 80%; 1 H NMR (500 MHz, DMSO-d6) δ 9.71 (s, 1H), 8.58 (d, J = 5.0 Hz, 1H), 8.25–8.22 (m, 2H),7.85–7.82 (m, 2H), 7.61–7.57 (m, 2H), 7.45 (d, J = 5.0 Hz, 1H), 7.34–7.30 (m,2H), 6.97 (tt, J = 7.5, 1.0 Hz, 1H), 3.68–3.53 (s, 2H), 3.51–3.33 (m, 6H),1.41 (s, 9H). ESI-HRMS m / z: calcd for C 26 H 30 O3N5 + [M + H] + , 460.2343; the measured value is 460.2378.

[0152] Synthesis of compound 15c: reactant 14c was selected to give compound 15c, yield: 75%; 1H NMR (500MHz, DMSO-d6) δ 9.86 (s, 1H), 8.61 (d, J = 5.0 Hz, 1H), 8.28– 8.24 (m, 2H), 7.97–7.93 (m, 2H), 7.68–7.64 (m, 4H), 7.61–7.58 (m, 2H), 7.47 (d, J = 5.0 Hz, 1H), 7.46–7.41 (m, 2H), 7.33–7.29 (m, 1H), 3.68–3.35 (m, 8H), 1.41 (s, 9H). ESI-HRMS m / z: calcd for C 32 H 34 O3N5 + [M + H] + , 536.2656; found, 536.2648.

[0153] Synthesis of compound 15d: Reactant selected 14d, to obtain compound 15d in yield: 85%; 1 H NMR (500MHz, DMSO-d6) δ 9.58 (s, 1H), 8.55 (d, J = 5.0 Hz, 1H), 8.24– 8.19 (m, 2H), 7.74–7.69 (m, 2H), 7.60–7.55 (m, 2H), 7.40 (d, J = 5.0 Hz, 1H), 7.17–7.13 (m, 2H), 3.66–3.33 (m, 8H), 2.44 (td, J = 10.0, 5.0 Hz, 1H), 1.81–1.74 (m, 4H), 1.73–1.66 (m, 1H), 1.41 (s, 9H), 1.39–1.30 (m, 4H), 1.27–1.21 (m, 1H). ESI-HRMS m / z: calcd for C 32 H 40 O3N5 + [M + H] + , 542.1326; found, 542.3120.

[0154] (2) Synthesis of compounds 16a-16d

[0155] The reaction scheme is:

[0156] ;

[0157] The compound 15a-15d prepared in step (1) 500 mg was dissolved in 9.0 mL of DCM, 3 mL of hydrogen chloride-1,4 dioxane solution was added, and the reaction was carried out at room temperature for 2 h. After the reaction was completed, the solid was collected by filtration and dried to obtain yellow solid compound 16a-16d.

[0158] Example 6 Synthesis of compounds A1-A24

[0159] The reaction scheme is as follows:

[0160] ;

[0161] The compound 14a-14d prepared in example 4 (0.2 mmol, 1.0 equiv) and the compound 3a-3f prepared in example 1 (0.22 mmol, 1.1 equiv) were dissolved in N,N-dimethylformamide, HATU (0.22 mmol, 1.1 equiv) and DIPEA (1.0 mmol, 5.0 equiv) were added, and the reaction was carried out at room temperature for 10 h. After the reaction was completed, water was added for quenching, and the organic phase was extracted with ethyl acetate, dried with anhydrous sodium sulfate, and concentrated. The compound A1-A24 was obtained by column chromatography.

[0162] The structural formula of the compound A1-A24 is as follows:

[0163]

[0164] .

[0165] Synthesis of compound A1: Compound 14a and compound 3a were reacted to obtain compound A1, yield: 43%; 1H NMR(400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.51 (s, 1H), 8.86 (t, J = 5.2 Hz, 1H), 8.52 (d, J = 5.2 Hz, 1H), 8.23 ​​(d, J = 8.0 Hz, 2H), 7.98 (d, J = 8.0 Hz, 2H),7.67 (d, J = 8.4 Hz, 2H), 7.59 (t, J = 8.0 Hz, 1H), 7.39 (d, J = 5.2 Hz, 1H),7.26 (d, J = 8.8 Hz, 1H), 7.03 (d, J = 7.2 Hz, 1H), 6.93 (d, J = 8.4 Hz, 2H), 6.90–6.84 (m, 1H), 5.06 (dd, J = 12.8, 5.2 Hz, 1H), 3.80–3.70 (m, 4H), 3.59–3.48 (m, 4H), 3.09–2.99 (m, 4H), 2.93–2.83 (m, 1H), 2.63–2.52 (m, 2H), 2.05–1.98 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 168.8, 167.4, 166.3,162.6, 160.4, 159.3, 146.4, 146.3, 139.3, 136.3, 136.1, 132.9, 132.3, 127.8(2×C), 126.8 (2×C), 120.4 (2×C), 117.3, 115.7 (2×C), 110.6, 109.3, 107.6,66.2 (2×C), 49.3 (2×C), 48.6, 41.3, 39.0, 31.0, 22.2. ESI-HRMS m / z: calculated value is C 36 H 34 O6N8Na + [M + Na] + , 697.2494; the measured value is 697.2500.

[0166] Synthesis of Compound A2: Compound 14a and Compound 3b were reacted to give Compound A2, yield: 56%; 1H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.51 (s, 1H), 8.71 (t, J = 5.6 Hz, 1H), 8.52 (d, J = 5.2 Hz, 1H), 8.27–8.21 (m, 2H), 8.00 (d, J = 8.4 Hz, 2H), 7.71–7.64 (m, 2H), 7.58 (dd, J = 8.4, 7.2 Hz, 1H), 7.39 (d, J = 5.2 Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 7.03 (d, J = 7.2 Hz, 1H), 6.97–6.89 (m, 2H), 6.79 (t, J = 6.0 Hz, 1H), 5.06 (dd, J = 12.8, 5.2 Hz, 1H), 3.78–3.71 (m, 4H), 3.45–3.38 (m, 4H), 3.08–3.01 (m, 4H), 2.93–2.91 (m, 1H), 2.62–2.51 (m, 2H), 2.07–1.98 (m, 1H), 1.88–1.78 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 168.9, 167.4, 165.9, 162.7, 160.4, 159.3, 146.3, 146.3, 139.2, 136.4, 136.3, 132.9, 132.3, 127.8 (2 x C), 126.8 (2 x C), 120.4 (2 x C), 117.2, 115.7 (2 x C), 110.5, 109.2, 107.6, 66.2 (2 x C), 49.3 (2 x C), 48.6, 39.0, 36.8, 31.0, 28.7, 22.2. ESI-HRMS m / z: calcd for C 37 H 36 O6N8Na + [M + Na] + , 711.2650; found 711.2653.

[0167] Synthesis of compound A3: Compound 14a and compound 3c were reacted to give compound A3 in yield of 51%; 1H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.47 (s, 1H), 8.62 (t, J = 5.5 Hz, 1H), 8.52 (d, J = 5.0 Hz, 1H), 8.22 (d, J = 8.0 Hz, 2H), 7.98 (d, J = 8.0 Hz, 2H), 7.67 (d, J = 8.5 Hz, 2H), 7.57 (dd, J = 8.5, 7.0 Hz, 1H), 7.38 (d, J = 5.0 Hz, 1H), 7.12 (d, J = 8.5 Hz, 1H), 7.01 (d, J = 7.0 Hz, 1H), 6.93 (d, J = 9.0 Hz, 2H), 6.58 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.5, 5.5 Hz, 1H), 3.78–3.71 (m, 4H), 3.44–3.29 (m, 4H), 3.09–3.29 (m, 4H), 2.94–2.82 (m, 1H), 2.63–2.51 (m, 2H), 2.06–1.99 (m, 1H), 1.70–1.58 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 173.0, 170.2, 169.0, 167.4, 165.7, 162.7, 160.4, 159.3, 146.5, 146.3, 139.2, 136.5, 136.4, 133.0, 132.3, 127.8 (2×C), 126.8 (2×C), 120.4 (2×C), 117.3, 115.7 (2×C), 110.5, 109.1, 107.6, 66.3 (2×C), 49.3 (2×C), 48.6, 41.6, 39.0, 31.0, 26.6, 26.3, 22.2. ESI-HRMS m / z: calculated for C 38 H 38 O6N8Na + [M + Na] + ,725.2807; found, 724.2803.

[0168] Synthesis of compound A4: Compound 14a and compound 3d were reacted to give compound A4 in yield of 40%; 1H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.47 (s, 1H), 8.59 (t, J = 5.5 Hz, 1H), 8.52 (d, J = 5.0 Hz, 1H), 8.24–8.18 (m, 2H), 8.01–7.95 (m, 2H), 7.70–7.63 (m, 2H), 7.57 (dd, J = 8.5, 7.0 Hz, 1H), 7.38 (d, J = 5.0 Hz, 1H), 7.11 (d, J = 8.5 Hz, 1H), 7.01 (d, J = 7.0 Hz, 1H), 6.96–6.89 (m, 2H), 6.55 (t, J = 6.0 Hz, 1H), 5.04 (dd, J = 12.5, 5.5 Hz, 1H), 3.77–3.71 (m, 4H), 3.32–3.28 (m, 4H), 3.08–3.00 (m, 4H), 2.91–2.83 (m, 1H), 2.61–2.51 (m, 2H), 2.04–1.99 (m, 1H), 1.66–1.56 (m, 4H), 1.46–1.39 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.4, 165.6, 162.7, 160.4, 159.3, 146.5, 146.3, 139.1, 136.5, 136.4, 132.9, 132.3, 127.7 (2 x C), 126.8 (2 x C), 120.4 (2 x C), 117.3, 115.7 (2 x C), 110.5, 109.0, 107.6, 66.3 (2 x C), 49.3 (2 x C), 48.6, 41.9, 39.0, 31.0, 28.9, 28.5, 23.9, 22.2. ESI-HRMS m / z: calcd for C 39 H 40 O6N8Na + [M + Na] + ,739.2963; found, 739.2969.

[0169] Synthesis of compound A5: Compound 14a and compound 3e were reacted to give compound A5 in yield of 60%; 1H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.47 (s, 1H), 8.58 (t, J = 5.5 Hz, 1H), 8.52 (d, J = 5.0 Hz, 1H), 8.24–8.18 (m, 2H), 8.01–7.95 (m, 2H), 7.69– 7.64 (m, 2H), 7.57 (dd, J = 8.5, 7.0 Hz, 1H), 7.38 (d, J = 5.0 Hz, 1H), 7.08 (d, J = 8.5 Hz, 1H), 7.01 (d, J = 7.0 Hz, 1H), 6.96–6.90 (m, 2H), 6.53 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.5, 5.5 Hz, 1H), 3.77–3.70 (m, 4H), 3.31–3.25 (m, 4H), 3.07–3.01 (m, 4H), 2.92–2.83 (m, 1H), 2.61–2.51 (m, 2H), 2.06–1.98 (m, 1H), 1.62–1.52 (m, 4H), 1.42–1.34 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.4, 165.6, 162.7, 160.4, 159.3, 146.5, 146.2, 139.1, 136.5, 136.3, 132.9, 132.2, 127.7 (2 x C), 126.8 (2 x C), 120.3 (2 x C), 117.2, 115.7 (2 x C), 110.4, 109.0, 107.6, 66.2 (2 x C), 49.3 (2 x C), 48.6, 41.8, 39.0, 31.0, 29.1, 28.7, 26.3, 26.1, 22.2. ESI-HRMS m / z: calcd for C 40 H 42 O6N8Na + [M +Na] + , 753.3120; found, 753.3126.

[0170] Synthesis of compound A6: Compound 14a and compound 3f were reacted to give compound A6 in yield of 43%; 1H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 9.50 (s, 1H), 8.65 (t, J = 5.2 Hz, 1H), 8.52 (d, J = 5.2 Hz, 1H), 8.21 (d, J = 8.4 Hz, 2H), 7.99 (d, J = 8.4 Hz, 2H), 7.70 - 7.63 (m, 2H), 7.54 (dd, J = 8.4, 7.2 Hz, 1H), 7.37 (d, J = 4.8 Hz, 1H), 7.10 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 7.2 Hz, 1H), 6.96 - 6.89 (m, 2H), 6.59 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.77 - 3.71 (m, 4H), 3.61 (t, J = 5.2 Hz, 2H), 3.60 - 3.53 (m, 6H), 3.47 - 3.41 (m, 4H), 3.08 - 3.00 (m, 4H), 2.92 - 2.82 (m, 1H), 2.62 - 2.51 (m, 2H), 2.06 - 1.96 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.3, 165.8, 162.6, 160.4, 159.3, 146.4, 146.3, 139.2, 136.3, 136.2, 132.9, 132.1, 127.7 (2 x C), 126.8 (2 x C), 120.4 (2 x C), 117.4, 115.7 (2 x C), 110.7, 109.3, 107.6, 69.7 (2 x C), 68.9 (2 x C), 66.2 (2 x C), 49.3 (2 x C), 48.6, 41.7, 39.0, 31.0, 22.2. ESI-HRMS m / z: calcd for C 40 H 42 O8N8Na + [M + Na] + , 785.3018; found 785.3034.

[0171] Synthesis of compound A7: Compound 14b and compound 3a were reacted to give compound A7 in yield of: 60%;1 H NMR (500 MHz, DMSO-d6) δ 11.09 (br, 1H), 9.73 (s, 1H), 9.00 (t, J = 5.5 Hz, 1H), 8.59 (d, J = 5.0 Hz, 1H), 8.27–8.23 (m, 2H), 8.05–8.01 (m, 2H), 7.86–7.81 (m, 2H), 7.59 (dd, J = 8.5, 7.0 Hz, 1H), 7.48 (d, J = 5.0 Hz, 1H), 7.34–7.27 (m, 3H), 7.02 (d, J = 7.0 Hz, 1H), 6.97 (tt, J = 7.5, 1.0 Hz, 1H), 6.86 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.5, 5.5 Hz, 1H), 3.57–3.48 (m, 4H), 2.92–2.84 (m, 1H), 2.61–2.51 (m, 2H), 2.04–1.99 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 173.0, 170.2, 168.8, 167.4, 166.3, 162.8, 160.3, 159.4, 146.4, 140.6, 139.2, 136.3, 136.2, 132.3, 128.6 (2×C), 127.9 (2×C), 126.9 (2×C), 121.6, 119.0 (2×C), 117.4, 110.7, 109.3, 108.4, 48.6, 41.4, 39.0, 31.0, 22.2. ESI-HRMS m / z: Calcd for C 32 H 28 O5N7 + [M + H] + , 590.2146; Found, 590.2141.

[0172] Synthesis of compound A8: Compound 14b and compound 3b were reacted to give compound A8 in yield of 54%; 1H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.75 (s, 1H), 8.73 (t, J = 5.6 Hz, 1H), 8.59 (d, J = 5.2 Hz, 1H), 8.29–8.23 (m, 2H), 8.02 (d, J = 8.4 Hz, 2H), 7.88–7.82 (m, 2H), 7.57 (dd, J = 8.4, 7.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 7.35–7.29 (m, 2H), 7.12 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 7.00–6.94 (m, 1H), 6.79 (t, J = 6.0 Hz, 1H), 5.07 (dd, J = 12.8, 5.2 Hz, 1H), 3.44–3.38 (m, 4H), 2.93–2.83 (m, 1H), 2.63–2.51 (m, 2H), 2.06–1.99 (m, 1H), 1.88–1.80 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 168.9, 167.4, 165.9, 162.8, 160.2, 159.3, 146.3, 140.6, 139.1, 136.5, 136.3, 132.3, 128.6 (2 x C), 127.8 (2 x C), 126.8 (2 x C), 121.5, 119.0 (2 x C), 117.2, 110.5, 109.2, 108.3, 48.6, 39.0, 36.8, 31.0, 28.7, 22.2. ESI-HRMS m / z: calcd for C 33 H 30 O5N7 + [M + H] + ,604.2303; found 604.2299.

[0173] Synthesis of compound A9: Compound 14b and compound 3c were reacted to give compound A9 in yield of 44%; 1H NMR(500 MHz, DMSO-d6) δ 11.09 (br, 1H), 9.73 (s, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.58 (d, J = 5.0 Hz, 1H), 8.26–8.21 (m, 2H), 8.05–8.01 (m, 2H), 7.86–7.81 (m,2H), 7.56 (dd, J = 8.5, 7.0 Hz, 1H), 7.47 (d, J = 5.0 Hz, 1H), 7.34–7.29 (m,2H), 7.13 (d, J = 8.5 Hz, 1H), 7.00 (d, J = 7.0 Hz, 1H), 6.97 (tt, J = 7.0,1.0 Hz, 1H), 6.58 (t, J = 6.0 Hz, 1H), 5.04 (dd, J = 13.0, 5.5 Hz, 1H), 3.35–3.31 (m, 4H), 2.92–2.84 (m, 1H), 2.60–2.51 (m, 2H), 2.05–1.99 (m, 1H), 1.67–1.59 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 173.0, 170.2, 169.0, 167.4, 165.6,162.8, 160.2, 159.4, 146.5, 140.6, 139.0, 136.5, 136.4, 132.3, 128.6 (2×C),127.9 (2×C), 126.8 (2×C), 121.5, 119.0 (2×C), 117.4, 110.5, 109.0, 108.4,48.6, 41.6, 38.9, 31.0, 26.5, 26.3, 22.2. ESI-HRMS m / z: The calculated value is C 34 H 32 O5N7 + [M+H] + , 618.2459; the measured value is 618.2453.

[0174] Synthesis of compound A10: Compound 14b and compound 3d were reacted to give compound A10, yield: 42%; 1HNMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 9.74 (s, 1H), 8.65 (t, J = 5.6 Hz, 1H), 8.59 (d, J = 5.2 Hz, 1H), 8.27–8.20 (m, 2H), 8.02–7.97 (m, 2H), 7.87–7.81 (m, 2H), 7.57 (dd, J = 8.4, 7.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 7.35–7.29 (m, 2H), 7.11 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 7.2 Hz, 1H), 6.97 (tt, J = 7.2, 1.2 Hz, 1H), 6.55 (t, J = 6.0 Hz, 1H), 5.04 (dd, J = 12.8, 5.4 Hz, 1H), 3.34–3.27 (t, 4H), 2.92–2.81 (m, 1H), 2.60–2.53 (m, 2H), 2.03–1.98 (m, 1H), 1.65–1.55 (m, 4H), 1.44–1.38 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.4, 165.6, 162.8, 160.3, 159.4, 146.5, 140.6, 139.0, 136.6, 136.4, 132.3, 128.6 (2 x C), 127.8 (2 x C), 126.8 (2 x C), 121.6, 119.0 (2 x C), 117.3, 110.5, 109.0, 108.4, 48.6, 41.9, 39.0, 31.0, 28.9, 28.5, 23.9, 22.2. ESI-HRMS m / z: calcd for C 35 H 34 O5N7 + [M + H] + , 632.2616; found 632.2616.

[0175] Synthesis of compound A11: Compound 14b and compound 3e were reacted to give compound A11 in yield of 47%; 1HNMR (400 MHz, DMSO-d6) δ 11.10 (br, 1H), 9.75 (s, 1H), 8.68 (t, J = 5.6 Hz, 1H), 8.58 (d, J = 5.2 Hz, 1H), 8.27–8.20 (m, 2H), 8.04–7.99 (m, 2H), 7.87–7.81 (m, 2H), 7.57 (dd, J = 8.4, 7.2 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 7.35–7.28 (m, 2H), 7.09 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 7.2 Hz, 1H), 6.99–6.94 (m, 1H), 6.55 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.33–2.25 (m, 4H), 2.92–2.82 (m, 1H), 2.62–2.52 (m, 2H), 2.06–1.99 (m, 1H), 1.62–1.52 (m, 4H), 1.42–1.33 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0,167.4, 165.5, 162.8, 160.2, 159.3, 146.5, 140.6, 138.9, 136.6, 136.4, 132.2, 128.6 (2×C), 127.8 (2×C), 126.8 (2×C), 121.5, 119.0 (2×C), 117.3, 110.4, 109.0, 108.3, 48.6, 41.8, 39.0, 31.0, 29.1, 28.7, 26.3, 26.1, 22.2. ESI-HRMS m / z: calcd for C 36 H 36 O5N7 + [M + H] + , 646.2772; found 646.2768.

[0176] Synthesis of compound A12: Compound 14b and compound 3f were reacted to give compound A12 in yield of 42%; 1HNMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.75 (s, 1H), 8.66 (t, J = 5.6 Hz, 1H), 8.58 (d, J = 5.2 Hz, 1H), 8.26–8.20 (m, 2H), 8.03–7.98 (m, 2H), 7.87–7.80 (m, 2H), 7.54 (dd, J = 8.4, 7.0 Hz, 1H), 7.46 (d, J = 5.2 Hz, 1H), 7.36–7.28 (m, 2H), 7.10 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 7.2 Hz, 1H), 6.97 (tt, J = 7.2, 1.2 Hz, 1H), 6.59 (t, J = 5.6 Hz, 1H), 5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.61 (t, J = 5.2 Hz, 2H), 3.60–3.54 (m, 6H), 3.48–3.41 (m, 4H), 2.92–2.81 (m, 1H), 2.62–2.51 (m, 2H), 2.05–1.97 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.3, 165.7, 162.7, 160.2, 159.3, 146.4, 140.6, 139.1, 136.3, 136.3, 132.1, 128.6 (2 x C), 127.8 (2 x C), 126.8 (2 x C), 121.5, 119.0 (2 x C), 117.4, 110.7, 109.3, 108.3, 69.7 (2 x C), 68.9 (2 x C), 48.6, 41.7, 39.0, 31.0, 22.2. ESI-HRMS m / z: calcd for C 36 H 36 O7N7 + [M + H] + , 678.2671; found 678.2671.

[0177] Synthesis of compound A13: Compound 14c and compound 3a were reacted to give compound A13 in yield of 46%; 1HNMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.87 (s, 1H), 8.84 (t, J = 5.5 Hz,1H), 8.62 (d, J = 5.0 Hz, 1H), 8.30–8.26 (m, 2H), 8.03–7.99 (m, 2H), 7.97–7.94 (m, 2H), 7.68–7.64 (m, 4H), 7.59 (dd, J = 8.5, 7.0 Hz, 1H), 7.51 (d, J =5.0 Hz, 1H), 7.44 (t, J = 8.0 Hz, 2H), 7.33–7.30 (m, 1H), 7.27 (d, J = 8.5Hz, 1H), 7.03 (d, J = 7.0 Hz, 1H), 6.87 (t, J = 6.0 Hz, 1H), 5.06 (dd, J =12.5, 5.5 Hz, 1H), 3.57–3.48 (m, 4H), 2.92–2.83 (m, 1H), 2.62–2.51 (m, 2H), 2.05–1.99 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 173.0, 170.2, 168.8, 167.4,166.4, 162.9, 160.2, 159.4, 146.4, 140.1, 140.0, 139.2, 136.3, 136.3, 133.2,132.3, 129.0 (2×C), 127.9 (2×C), 127.0 (2×C), 126.9 (2×C), 126.8, 126.2(2×C), 119.3 (2×C), 117.3, 110.7, 109.4, 108.6, 48.6, 41.4, 39.0, 31.1,22.2. ESI-HRMS m / z: calculated for C 38 H 32 O5N7 + [M + H] + , 666.2459; the measured value is 666.2449.

[0178] Synthesis of compound A14: Compound 14c and compound 3b were reacted to give compound A14, yield: 47%; 1HNMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.91 (s, 1H), 8.73 (t, J = 5.6 Hz,1H), 8.62 (d, J = 5.2 Hz, 1H), 8.32–8.25 (m, 2H), 8.05–8.00 (m, 2H), 7.98–7.93 (m, 2H), 7.69–7.63 (m, 4H), 7.58 (dd, J = 8.4, 7.2 Hz, 1H), 7.51 (d, J =5.2 Hz, 1H), 7.44 (t, J = 7.6 Hz, 2H), 7.34–7.29 (m, 1H), 7.13 (d, J = 8.4Hz, 1H), 7.03 (d, J = 7.2 Hz, 1H), 6.80 (t, J = 6.0 Hz, 1H), 5.06 (dd, J =12.8, 5.2 Hz, 1H), 3.44–3.38 (m, 4H), 2.93–2.83 (m, 1H), 2.62–2.51 (m, 2H), 2.06–1.98 (m, 1H), 1.89–1.78 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9,170.2, 168.9, 167.4, 165.9, 162.9, 160.2, 159.4, 146.3, 140.1, 140.0, 139.0,136.5, 136.3, 133.1, 132.3, 128.9 (2×C), 127.8 (2×C), 126.9 (2×C), 126.8(2×C), 126.7, 126.1 (2×C), 119.2 (2×C), 117.2, 110.5, 109.2, 108.5, 48.6,39.0, 36.8, 31.0, 28.7, 22.2. ESI-HRMS m / z: calculated for C 39 H 34 O5N7 + [M + H] + ,680.2616; the measured value is 680.2612.

[0179] Synthesis of compound A15: Compound 14c and compound 3c were reacted to give compound A15, yield: 34%; 1HNMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.90 (s, 1H), 8.67 (t, J = 5.6 Hz,1H), 8.62 (d, J = 5.2 Hz, 1H), 8.31–8.24 (m, 2H), 8.01 (d, J = 8.4 Hz, 2H), 7.97–7.93 (m, 2H), 7.67 (dd, J = 5.2, 1.6 Hz, 2H), 7.65–7.64 (m, 1H), 7.57 (dd, J = 8.4, 7.2 Hz, 1H), 7.50 (d, J = 5.2 Hz, 1H), 7.44 (t, J = 7.6 Hz,2H), 7.34–7.29 (m, 1H), 7.28–7.25 (m, 1H), 7.12 (d, J = 8.4 Hz, 1H), 7.01 (d,J = 7.2 Hz, 1H), 6.60 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.8, 5.4 Hz, 1H),3.36–3.31 (m, 4H), 2.93–2.82 (m, 1H), 2.62–2.51 (m, 2H), 2.06–1.98 (m, 1H),1.68–1.59 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.4,165.6, 162.9, 160.2, 159.5, 159.4, 146.4, 140.1, 140.0, 139.0, 136.7, 136.6,136.3, 133.1, 132.3, 130.2, 128.9 (2×C), 127.8, 126.9, 126.8, 126.7, 126.1(2×C), 120.0, 119.2, 117.3, 110.4, 109.0, 108.5, 48.6, 41.6, 39.0, 31.0,26.6, 26.3, 22.2. ESI-HRMS m / z: Calculated for C 40 H 36 O5N7 + [M + H] + , 694.2772; the measured value is 694.2767.

[0180] Synthesis of compound A16: Compound 14c and compound 3d were reacted to give compound A16, yield: 35%; 1 HNMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.90 (s, 1H), 8.63 (d, J = 5.2 Hz,2H), 8.30–8.24 (m, 2H), 8.02–7.99 (m, 2H), 7.96 (dd, J = 8.8, 2.0 Hz, 2H),7.67 (d, J = 1.2 Hz, 1H), 7.66–7.64 (m, 2H), 7.57 (dd, J = 8.4, 7.2 Hz, 1H),7.50 (d, J = 5.2 Hz, 1H), 7.47–7.42 (m, 2H), 7.34–7.30 (m, 1H), 7.30–7.24 (m,1H), 7.10 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 7.2 Hz, 1H), 6.56 (t, J = 6.0 Hz,1H), 5.05 (dd, J = 12.8, 5.4 Hz, 1H), 3.34–3.27 (m, 4H), 2.93–2.82 (m, 1H), 2.62–2.51 (m, 2H), 2.05–1.96 (m, 1H), 1.67–1.55 (m, 4H), 1.46–1.36 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.4, 165.6, 162.9, 160.2,159.5, 159.4, 146.4, 140.1, 140.0, 138.9, 136.6, 136.3, 133.1, 132.2, 130.2,128.9 (2×C), 127.8, 126.9, 126.8, 126.7, 126.1 (2×C), 120.0, 119.2 (2×C),117.3, 110.4, 109.0, 108.5, 48.6, 41.8, 39.0, 31.0, 28.9, 28.5, 23.9, 22.2. ESI-HRMS m / z: Calculated for C 41 H 38 O5N7 + [M + H] +, 708.2929; the measured value is 708.2962.

[0181] Synthesis of compound A17: Compound 14c and compound 3e were reacted to give compound A17, yield: 59%; 1 HNMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.90 (s, 1H), 8.62 (d, J = 5.2 Hz,2H), 8.27 (d, J = 8.4 Hz, 2H), 8.04–7.99 (m, 2H), 7.98–7.93 (m, 2H), 7.69–7.62 (m, 4H), 7.56 (dd, J = 8.4, 7.2 Hz, 1H), 7.50 (d, J = 5.2 Hz, 1H), 7.44(t, J = 7.6 Hz, 2H), 7.34–7.28 (m, 1H), 7.08 (d, J = 8.4 Hz, 1H), 7.01 (d, J= 7.2 Hz, 1H), 6.55 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.8, 5.4 Hz, 1H), 3.33–3.24 (m, 4H), 2.93–2.83 (m, 1H), 2.63–2.51 (m, 2H), 2.06–1.99 (m, 1H), 1.63–1.52 (m, 4H), 1.42–1.34 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9,170.2, 169.0, 167.4, 165.6, 162.9, 160.2, 159.3, 146.4, 140.1, 140.0, 138.9,136.6, 136.3, 133.1, 132.2, 128.9 (2×C), 127.8 (2×C), 126.9 (2×C), 126.8(2×C), 126.7, 126.1 (2×C), 119.2 (2×C), 117.2, 110.4, 109.0, 108.5, 48.6,41.8, 39.0, 31.0, 29.1, 28.7, 26.3, 26.2, 22.2. ESI-HRMS m / z: Calculated for C 42 H 40 O5N7 + [M + H] +, 722.3085; the measured value is 722.3084.

[0182] Synthesis of compound A18: Compound 14c and compound 3f were reacted to give compound A18, yield: 54%; 1 HNMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.90 (s, 1H), 8.67 (t, J = 5.6 Hz,1H), 8.61 (d, J = 5.2 Hz, 1H), 8.28–8.22 (m, 2H), 8.05–7.99 (m, 2H), 7.98–7.92 (m, 2H), 7.69–7.63 (m, 4H), 7.54 (dd, J = 8.4, 7.2 Hz, 1H), 7.48 (d, J =5.2 Hz, 1H), 7.44 (t, J = 7.6 Hz, 2H), 7.33–7.28 (m, 1H), 7.10 (d, J = 8.4Hz, 1H), 7.00 (d, J = 7.2 Hz, 1H), 6.59 (t, J = 6.0 Hz, 1H), 5.05 (dd, J =12.8, 5.4 Hz, 1H), 3.61 (t, J = 5.4 Hz, 2H), 3.60–3.55 (m, 6H), 3.48–3.42 (m,4H), 2.93–2.80 (m, 1H), 2.61–2.51 (m, 2H), 2.04–1.97 (m, 1H). 13 C NMR (101MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.4, 165.8, 162.8, 160.2, 159.4,146.4, 140.1, 140.0, 139.1, 136.3, 136.2, 133.1, 132.1, 128.9 (2×C), 127.8(2×C), 126.9 (2×C), 126.8 (2×C), 126.7, 126.1 (2×C), 119.2 (2×C), 117.5,110.7, 109.3, 108.5, 69.7 (2×C), 68.9 (2×C), 48.6, 41.7, 39.0, 31.0, 22.2. ESI-HRMS m / z: calculated for C 42 H 40 O7N7 +[M + H] + Found 754.2981.

[0183] Synthesis of compound A19: Compound 14d and compound 3a were reacted to give compound A19 in yield: 60%; 1 HNMR (500 MHz, DMSO-d6) δ 11.11 (s, 1H), 9.63 (s, 1H), 8.84 (t, J = 5.5 Hz,1H), 8.56 (d, J = 5.0 Hz, 1H), 8.24 (d, J = 8.0 Hz, 2H), 7.99 (d, J = 8.0 Hz,2H), 7.71 (d, J = 8.0 Hz, 2H), 7.59 (t, J = 8.0 Hz, 1H), 7.44 (d, J = 5.0 Hz,1H), 7.26 (d, J = 8.5 Hz, 1H), 7.15 (d, J = 8.0 Hz, 2H), 7.03 (d, J = 7.0 Hz,1H), 6.89 (d, J = 6.0 Hz, 1H), 5.06 (dd, J = 12.5, 5.5 Hz, 1H), 3.58–3.47 (m,4H), 2.94–2.93 (m, 1H), 2.62-2.52 (m, 2H), 2.46–2.39 (m, 1H), 2.07–1.98 (m,1H), 1.78 (d, J = 10.0 Hz, 4H), 1.72–1.66 (m, 1H), 1.41–1.31 (m, 4H), 1.26–1.19 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 173.0, 170.2, 168.8, 167.4, 166.4,162.7, 160.4, 159.4, 146.5, 141.0, 139.3, 138.3, 136.4, 136.2, 132.3, 127.8(2×C), 126.9 (2×C), 126.7 (2×C), 119.2 (2×C), 117.3, 110.7, 109.4, 108.1,48.6, 43.3, 41.4, 39.0, 34.3 (2×C), 31.1, 26.5 (2×C), 25.8, 22.3. ESI-HRMS m / z: Calcd for C 38 H38 O5N7 + [M + H] + Found 672.2924.

[0184] Synthesis of compound A20: Compound 14d and compound 3b were reacted to give compound A20 in yield: 65%; 1 HNMR (500 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.63 (s, 1H), 8.71 (t, J = 5.5 Hz,1H), 8.56 (d, J = 5.0 Hz, 1H), 8.25 (d, J = 8.0 Hz, 2H), 8.01 (d, J = 8.0 Hz,2H), 7.72 (d, J = 8.0 Hz, 2H), 7.58 (t, J = 8.0 Hz, 1H), 7.44 (d, J = 5.0 Hz,1H), 7.18-7.11 (m, 3H), 7.03 (d, J = 7.0 Hz, 1H), 6.78 (t, J = 6.0 Hz, 1H),5.06 (dd, J = 13.0, 5.5 Hz, 1H), 3.45–3,37 (m, 4H), 2.92–2.83 (m, 1H), 2.61–2.52 (m, 2H), 2.46–2.39 (m, 1H), 2.05–1.99 (m, 1H), 1.88–1.81 (m, 2H), 1.78(d, J = 10.0 Hz, 4H), 1.71–1.66 (m, 1H), 1.42–1.31 (m, 4H), 1.27–1.21 (m,1H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 168.9, 167.4, 165.9, 162.7,160.3, 159.3, 146.3, 140.9, 139.1, 138.3, 136.4, 136.3, 132.3, 127.8 (2×C),126.8 (2×C), 126.6 (2×C), 119.2 (2×C), 117.2, 110.5, 109.2, 108.0, 48.6,43.2, 39.0, 36.8, 34.2 (2×C), 31.0, 28.7, 26.5 (2×C), 25.7, 22.2. ESI-HRMS m / z: calculated 672.2929. Found 672.2924.39 H 40 O5N7 + [M + H] + , 686.3085; Found, 686.3081.

[0185] Synthesis of compound A21 : Compound 14d and compound 3c were reacted to give compound A21 in yield of 42%; 1 HNMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.64 (s, 1H), 8.65 (t, J = 5.6 Hz,1H), 8.56 (d, J = 5.2 Hz, 1H), 8.23 (d, J = 8.0 Hz, 2H), 7.99 (d, J = 8.0 Hz,2H), 7.71 (d, J = 8.0 Hz, 2H), 7.57 (t, J = 8.0 Hz, 1H), 7.43 (d, J = 5.2 Hz,1H), 7.16 (d, J = 8.2 Hz, 2H), 7.14–6.98 (m, 2H), 6.60 (t, J = 6.4 Hz, 1H),5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.36–3.31 (m, 4H), 2.94–2.81 (m, 1H), 2.62–2.51 (m, 2H), 2.47–2.40 (m, 1H), 2.06–1.98 (m, 1H), 1.78 (d, J = 9.6 Hz, 4H),1.73–1.68 (m, 1H), 1.67–1.57 (m, 4H), 1.44–1.31 (m, 4H), 1.26–1.22 (m, 1H). 13C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.4, 165.7, 162.8, 160.3,159.3, 146.4, 140.9, 139.1, 138.3, 136.5, 136.3, 132.3, 127.8 (2×C), 126.8(2×C), 126.7 (2×C), 119.2 (2×C), 117.3, 110.5, 109.1, 108.0, 48.6, 43.2,41.6, 39.0, 34.2 (2×C), 31.0, 26.6, 26.5 (2×C), 26.3, 25.7, 22.2. ESI-HRMS m / z: calcd for C 40 H 42 O5N7 + [M + H] + , 700.3242; found, 700.3235.

[0186] Synthesis of compound A22: Compound 14d and compound 3d were reacted to give compound A22 in yield of 37%; 1 HNMR (500 MHz, DMSO-d6) δ 11.11 (s, 1H), 9.63 (s, 1H), 8.66–8.52 (m, 2H), 8.23(d, J = 8.0 Hz, 2H), 7.99 (d, J = 8.0 Hz, 2H), 7.72 (d, J = 8.0 Hz, 2H), 7.57(t, J = 8.0 Hz, 1H), 7.46–7.38 (m, 1H), 7.15 (d, J = 8.5 Hz, 2H), 7.10 (d, J= 8.5 Hz, 1H), 7.01 (d, J = 6.5 Hz, 1H), 6.60–6.48 (m, 1H), 5.05 (dd, J =13.0, 5.5 Hz, 1H), 3.33–3.23 (m, 4H), 2.91–2.82 (m, 1H), 2.64–2.52 (m, 2H),2.45–2.39 (m, 1H), 2.05–1.96 (m, 1H), 1.78 (d, J = 10.0 Hz, 4H), 1.71–1.67(m, 1H), 1.65–1.53 (m, 4H), 1.44–1.31 (m, 6H), 1.27–1.21 (m, 1H).13 C NMR (101 MHz, DMSO-d6) δ 173.0, 170.3, 169.1, 167.4, 165.7, 162.8, 160.4, 159.4, 146.5, 141.0, 139.1, 138.3, 136.6, 136.4, 132.3, 127.8 (2 x C), 126.9 (2 x C), 126.7 (2 x C), 119.3 (2 x C), 117.3, 110.5, 109.1, 108.1, 48.6, 43.3, 41.9, 39.0, 34.3 (2 x C), 31.1, 28.9, 28.5, 26.5 (2 x C), 25.8, 23.9, 22.3. ESI-HRMS m / z: calcd for C 41 H 44 O5N7 + [M + H] + , 714.3398; found 714.3392.

[0187] Synthesis of compound A23: Compound 14d and compound 3e were reacted to give compound A23 in yield of 51%; 1HNMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.64 (s, 1H), 8.60 (t, J = 5.6 Hz,1H), 8.55 (d, J = 5.2 Hz, 1H), 8.25–8.20 (m, 2H), 8.00 (s, 1H), 7.98 (d, J =1.6 Hz, 1H), 7.74–7.69 (m, 2H), 7.57 (dd, J = 8.4, 7.2 Hz, 1H), 7.43 (d, J =5.2 Hz, 1H), 7.18–7.13 (m, 2H), 7.08 (d, J = 8.8 Hz, 1H), 7.01 (d, J = 7.2Hz, 1H), 6.55 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.32–3.25(m, 4H), 2.92–2.82 (m, 1H), 2.62–2.51 (m, 2H), 2.47–2.40 (m, 1H), 2.06–1.98 (m, 1H), 1.78 (d, J = 9.2 Hz, 4H), 1.72–1.66 (m, 1H), 1.62–1.52 (m, 4H), 1.43–1.32 (m, 8H), 1.28–1.21 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9,170.2, 169.0, 167.4, 165.6, 162.8, 160.3, 159.3, 146.5, 140.9, 139.0, 138.3,136.6, 136.4, 132.2, 127.8 (2×C), 126.8 (2×C), 126.6 (2×C), 119.2 (2×C), 117.2, 110.4, 109.0, 108.0, 48.6, 43.2, 41.8, 39.0, 34.2 (2×C), 31.0, 29.1,28.7, 26.5 (2×C), 26.3, 26.2, 25.7, 22.2. ESI-HRMS m / z: calculated for C 42 H 46 O5N7 + [M+H] + , 728.3555; the measured value is 728.3549.

[0188] Synthesis of compound A24: Compound 14d and compound 3f were reacted to give compound A24, yield: 51%; 1 HNMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.63 (s, 1H), 8.65 (t, J = 5.6 Hz,1H), 8.55 (d, J = 5.2 Hz, 1H), 8.22 (d, J = 8.4 Hz, 2H), 8.00 (d, J = 8.4 Hz, 2H), 7.71 (dd, J = 8.8, 2.8 Hz, 2H), 7.54 (dd, J = 8.4, 7.2 Hz, 1H), 7.42 (d,J = 5.2 Hz, 1H), 7.18–7.12 (m, 2H), 7.09 (d, J = 8.4 Hz, 1H), 7.01 (d, J =7.2 Hz, 1H), 6.59 (d, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.61(t, J = 5.2 Hz, 2H), 3.59–3.54 (m, 6H), 3.47–3.41 (m, 4H), 2.91–2.82 (m, 1H), 2.61–2.51 (m, 2H), 2.46–2.39 (m, 1H), 2.05–1.97 (m, 1H), 1.77 (d, J = 9.4 Hz,4H), 1.72–1.65 (m, 1H), 1.42–1.29 (m, 4H), 1.27–1.20 (m, 1H). 13 C NMR (101MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 167.3, 165.8, 162.7, 160.3, 159.3,146.4, 140.9, 139.1, 138.3, 136.3, 132.1, 127.8 (2×C), 126.8 (2×C), 126.7(2×C), 119.2 (2×C), 117.4, 114.7, 110.7, 109.3, 108.0, 69.7 (2×C), 68.9 (2×C), 48.6, 43.2, 41.7, 39.0, 34.2 (2×C), 31.0, 26.5 (2×C), 25.7, 22.2.ESI-HRMS m / z: calculated for C 42H 46 O7N7 + [M + H] + 760.3453; found 760.3449.

[0189] Example 7 Synthesis of compounds B1-B4

[0190] The reaction scheme is as follows:

[0191] ;

[0192] The compound 14a-14d prepared in Example 4 (0.2 mmol, 1.0 equiv) and compound 5a prepared in Example 2 (0.22 mmol, 1.1 equiv) were dissolved in N,N- dimethylformamide, HATU (0.22 mmol, 1.1 equiv) and DIPEA (1.0 mmol, 5.0 equiv) were added and reacted at room temperature for 10 h. After the reaction was completed, water was added for quenching, the organic phase was extracted with ethyl acetate, dried over anhydrous sodium sulfate, the organic phase was concentrated, and compounds B1-B4 were obtained by flash column chromatography.

[0193] The structural formula of compounds B1-B4 is as follows:

[0194] .

[0195] Synthesis of compound B1: compound 14a and compound 5a were reacted to obtain compound B1, yield: 56%; 1H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.48 (s, 1H), 8.50 (d, J = 5.2 Hz, 1H), 8.22–8.14 (m, 2H), 7.78 (t, J = 6.0 Hz, 1H), 7.69–7.63 (m, 2H), 7.55 (dd, J = 8.4, 7.2 Hz, 1H), 7.53–7.49 (m, 2H), 7.33 (d, J = 4.8 Hz, 1H), 7.10 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.95–6.90 (m, 2H), 6.58 (t, J = 6.0 Hz, 1H), 5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.76–3.71 (m, 4H), 3.71–3.61 (m, 2H), 3.59 (t, J = 5.2 Hz, 2H), 3.55–3.50 (m, 4H), 3.45–3.41 (m, 4H), 3.38–3.37 (m, 4H), 3.27–3.22 (m, 2H), 3.07–3.01 (m, 4H), 2.95 (s, 2H), 2.92–2.82 (m, 1H), 2.61–2.52 (m, 2H), 2.45–2.35 (m, 2H), 2.04–1.97 (m, 1H). 13 C NMR (101MHz, DMSO-d6) δ 172.9, 170.2, 169.1, 169.0, 168.5, 167.4, 162.7, 160.4, 159.3, 146.4, 146.2, 137.9, 137.8, 136.3, 132.9, 132.1, 127.5 (2×C), 127.0(2×C), 120.3 (2×C), 117.4, 115.7 (2×C), 110.8, 109.3, 107.4, 69.8, 69.6, 69.1, 68.9, 66.2 (2×C), 60.9, 53.0, 52.4, 49.3 (2×C), 48.6, 47.2, 41.7, 41.6, 38.2, 31.0, 22.2. ESI-HRMS m / z: calculated 1049.3802. Found 1049.3800. 46 H 52 O9N 10 Na+ [M + Na] + found 911.3817.

[0196] Synthesis of compound B2: reaction of compound 14b and compound 5a gave compound B2 in yield of 49%; 1 H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.70 (s, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.23-8.18 (m, 2H), 7.85-7.81 (m, 2H), 7.79 (t, J = 6.0 Hz, 1H), 7.57-7.54 (m, 1H), 7.53 (d, J = 8.0 Hz, 2H), 7.42 (d, J = 5.0 Hz, 1H), 7.34-7.29 (m, 2H), 7.10 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 7.0 Hz, 1H), 6.97 (tt, J = 7.5, 1.0 Hz, 1H), 6.57 (t, J = 6.0 Hz, 1H), 5.04 (dd, J = 13.0, 5.5 Hz, 1H), 3.73-3.61 (m, 2H), 3.59 (t, J = 5.5 Hz, 2H), 3.56-3.50 (m, 4H), 3.46-3.37 (m, 6H), 3.35-3.32 (m, 2H), 3.27-3.22 (m, 2H), 2.96 (s, 2H), 2.92-2.83 (m, 1H), 2.61-2.52 (m, 2H), 2.46-2.37 (s, 2H), 2.04-1.99 (m, 1H). 13C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.1, 169.0, 168.4, 167.4, 162.8, 160.2, 159.4, 146.4, 140.6, 138.1, 137.6, 136.3, 132.1, 128.6 (2 x C), 127.6 (2 x C), 127.0 (2 x C), 121.5, 119.0 (2 x C), 117.5, 110.8, 109.3, 108.2, 69.8, 69.6, 69.0, 68.9, 60.9, 53.9, 52.3, 48.6, 47.1, 41.7, 41.5, 38.2, 31.0, 22.2. ESI-HRMS m / z: calcd for C 42 H 46 O8N9 + [M + H] + 804.3464; found 804.3458.

[0197] Synthesis of compound B3: Compound 14c and compound 5a were reacted to give compound B3 in the yield of 38%; 1H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.88 (s, 1H), 8.60 (d, J = 5.2 Hz, 1H), 8.26–8.21 (m, 2H), 7.97–7.92 (m, 2H), 7.79 (t, J = 6.0 Hz, 1H), 7.70–7.61 (m, 4H), 7.58–7.52 (m, 3H), 7.48–7.39 (m, 3H), 7.34–7.28 (m, 1H), 7.10 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.58 (t, J = 5.8 Hz, 1H), 5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.74-3.61 (m, 2H), 3.59 (t, J = 5.2 Hz, 2H), 3.56–3.49 (m, 4H), 3.43 (t, J = 5.6 Hz, 6H), 3.37–3.33 (m, 2H), 3.25 (q, J = 5.6 Hz, 2H), 2.96 (s, 2H), 2.92–2.81 (m, 1H), 2.61–2.52 (m, 2H), 2.46–2.37 (m, 2H), 2.04–1.97 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.1, 169.0, 168.4, 167.3, 162.9, 160.2, 159.4, 146.4, 140.1, 140.0, 138.1, 137.6, 136.3, 133.1, 132.1, 128.9 (2×C), 127.6 (2×C), 127.1 (2×C), 126.8 (2×C), 126.7, 126.1 (2×C), 119.2 (2×C), 117.4, 110.8, 109.3, 108.3, 69.8, 69.6, 69.0, 68.8, 60.9, 52.9, 52.4, 48.6, 47.2, 41.6, 41.5, 38.2, 31.0, 22.2. ESI-HRMS m / z: Calculated 1049.3832. Found 1049.3830. 48 H 50 O8N9 + [M + H] +Calcd: 880.3777; Found: 880.3777.

[0198] Synthesis of compound B4: Reaction of compound 14d and compound 5a gave compound B4 in yield of 37%; 1 H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.62 (s, 1H), 8.54 (d, J = 5.2 Hz, 1H), 8.22 - 8.17 (m, 2H), 7.78 (t, J = 5.6 Hz, 1H), 7.74 - 7.69 (m, 2H), 7.58 - 7.51 (m, 3H), 7.38 (d, J = 5.2 Hz, 1H), 7.17 - 7.13 (m, 2H), 7.10 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.59 (t, J = 5.6 Hz, 1H), 5.05 (dd, J = 12.8, 5.2 Hz, 1H), 3.74 - 3.61 (m, 2H), 3.59 (t, J = 5.2 Hz, 2H), 3.56 - 3.50 (m, 4H), 3.48 - 3.38 (m, 6H), 3.37 - 3.31 (m, 2H), 3.25 (q, J = 5.6 Hz, 2H), 2.95 (s, 2H), 2.92 - 2.82 (m, 1H), 2.61 - 2.52 (m, 2H), 2.47 - 2.38 (m, 3H), 2.05 - 1.97 (m, 1H), 1.77 (d, J = 9.2 Hz, 4H), 1.72 - 1.65 (m, 1H), 1.43 - 1.29 (m, 4H), 1.26 - 1.20 (m, 1H). 13C NMR (101 MHz, DMSO-d6) δ 172.9, 170.2, 169.0, 169.0, 168.4, 167.3, 162.8, 160.3, 159.3, 146.4, 140.9, 138.3, 138.0, 137.7, 136.3, 132.1, 127.5 (2 x C), 127.0 (2 x C), 126.7 (2 x C), 119.1 (2 x C), 117.4, 110.8, 109.3, 107.8, 69.8, 69.6, 69.1, 68.8, 60.9, 53.1, 52.3, 48.6, 47.1, 43.2, 41.6, 41.5, 38.2, 34.2 (2 x C), 31.0, 26.5 (2 x C), 25.7, 22.2. ESI-HRMS m / z: calcd for C 48 H 56 O8N9 + [M + H] + , 886.4246; found 886.4240.

[0199] Example 8. Synthesis of compounds C1-C8

[0200] The reaction scheme is as follows:

[0201] ;

[0202] The compound 8a-8b prepared in Example 3 (0.2 mmol, 1.0 equiv) was dissolved in acetonitrile, and compound 16a-16d prepared in Example 5 (0.2 mmol, 1.0 equiv) and glacial acetic acid were added, and the reaction was carried out at room temperature for 1 h, then sodium triacetoxyborohydride (NaBH(OAc)3, 0.6 mmol, 3.0 equiv) was added, and the reaction was carried out at room temperature for 16 h, after the reaction was completed, the crude product was concentrated under vacuum, then white solid compound C1-C8 was obtained by flash column chromatography.

[0203] The structural formula of compound C1-C8 is as follows:

[0204]

[0205] .

[0206] Synthesis of compound C1: compound 16a and compound 8a were reacted to obtain compound C1, yield: 30%; 1H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.49 (s, 1H), 8.51 (d, J = 5.2 Hz, 1H), 8.23-8.16 (m, 2H), 7.80 (dd, J = 8.4, 7.2 Hz, 1H), 7.71-7.63 (m, 2H), 7.56-7.49 (m, 3H), 7.43 (d, J = 7.2 Hz, 1H), 7.36 (d, J = 5.2 Hz, 1H), 6.97-6.89 (m, 2H), 5.08 (dd, J = 12.8, 5.4 Hz, 1H), 4.20 (t, J = 6.4 Hz, 2H), 3.76-3.71 (m, 4H), 3.70-3.56 (m, 2H), 3.35-3.29 (s, 2H), 3.09-2.98 (m, 4H), 2.93-2.83 (m, 1H), 2.62-2.51 (m, 2H), 2.49-2.41 (m, 2H), 2.40-2.24 (m, 4H), 2.05-1.97 (m, 1H), 1.81-1.73 (m, 2H), 1.56-1.42 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.0, 168.4, 166.9, 165.4, 162.7, 160.4, 159.3, 156.0, 146.2, 138.0, 137.8, 137.1, 133.3, 132.9, 127.5 (2 x C), 127.0 (2 x C), 120.3 (2 x C), 119.8, 116.2, 115.7 (2 x C), 115.2, 107.4, 68.8, 66.2 (2 x C), 57.5, 53.0, 52.4, 49.3 (2 x C), 48.8, 47.2, 41.5, 31.0, 28.3, 25.8, 23.3, 22.0. ESI-HRMS m / z: Calcd for C 43 H 47 O7N8 + [M + H] + 787.3562; found 787.3545.

[0207] Synthesis of compound C2: Compound 16a and compound 8b were reacted to give compound C2 in yield of: 36%;1 H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.50 (s, 1H), 8.51 (d, J = 5.2 Hz, 1H), 8.22–8.18 (m, 2H), 7.80 (dd, J = 8.4, 7.2 Hz, 1H), 7.69–7.64 (m, 2H), 7.55–7.52 (m, 2H), 7.50 (d, J = 8.4 Hz, 1H), 7.43 (d, J = 7.2 Hz, 1H), 7.36 (d, J= 5.2 Hz, 1H), 6.95–6.91 (m, 2H), 5.08 (dd, J = 12.8, 5.2 Hz, 1H), 4.19 (t, J= 6.4 Hz, 2H), 3.76–3.72 (m, 4H), 3.65–3.59 (m, 2H), 3.34–3.28 (m, 2H), 3.06–3.02 (m, 4H), 2.92–2.83 (m, 1H), 1.37–1.32 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.0, 168.3, 166.9, 165.4,162.7, 160.4, 159.3, 156.1, 146.2, 138.1, 137.7, 137.1, 133.3, 132.9, 127.5(2×C), 127.0 (2×C), 120.3 (2×C), 119.8, 116.2, 115.7 (2×C), 115.2, 107.4,68.8, 66.2 (2×C), 57.6, 53.0, 52.5, 49.3 (2×C), 48.8, 47.2, 41.8, 31.0,28.4, 26.6, 26.2, 25.3, 22.0. ESI-HRMS m / z: Calculated for C 44 H 49 O7N8 + [M + H] + ,801.3719; the measured value is 801.3722.

[0208] Synthesis of compound C3: Compound 16b and compound 8a were reacted to give compound C3 in yield of 37%; 1 H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.74 (br, 1H), 8.58 (d, J = 5.2 Hz, 1H), 8.24–8.19 (m, 2H), 7.86–7.82 (m, 2H), 7.80 (dd, J = 8.4, 7.2 Hz, 1H), 7.57–7.53 (m, 2H), 7.50 (d, J = 8.4 Hz, 1H), 7.44 (dd, J = 6.0, 4.0 Hz, 2H), 7.35–7.29 (m, 2H), 6.97 (tt, J = 7.2, 1.2 Hz, 1H), 5.08 (dd, J = 12.8, 5.2 Hz, 1H), 4.19 (t, J = 6.4 Hz, 2H), 3.70–3.56 (m, 2H), 3.36–3.28 (m, 2H), 2.93–2.82 (m, 1H), 2.61–2.51 (m, 2H), 2.46–2.38 (m, 2H), 2.37–2.26 (m, 4H), 2.05–1.98 (m, 1H), 1.80–1.72 (m, 2H), 1.54–1.43 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.1, 168.4, 166.9, 165.4, 162.9, 160.3, 159.3, 156.1, 140.6, 138.2, 137.6, 137.1, 133.3, 128.6 (2 x C), 127.6 (2 x C), 127.1 (2 x C), 121.5, 119.8, 119.0 (2 x C), 116.2, 115.2, 108.2, 68.8, 57.6, 53.0, 52.5, 48.8, 47.3, 41.6, 31.0, 28.3, 25.8, 23.3, 22.1. ESI-HRMS m / z: calcd for C 39 H 40 O6N7 + [M + H] + ,702.3035; found, 702.3030.

[0209] Synthesis of compound C4: Compound 16b and compound 8b were reacted to give compound C4 in yield: 33%; 1 H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.73 (s, 1H), 8.58 (d, J = 5.2 Hz, 1H), 8.24-8.20 (m, 2H), 7.86-7.82 (m, 2H), 7.82-7.77 (m, 1H), 7.56-7.53 (m, 2H), 7.50 (d, J = 8.4 Hz, 1H), 7.44 (dd, J = 6.0, 4.0 Hz, 2H), 7.35-7.28 (m, 2H), 6.97 (tt, J = 7.2, 1.2 Hz, 1H), 5.07 (dd, J = 12.8, 5.2 Hz, 1H), 4.18 (t, J = 6.3 Hz, 2H), 3.65-3.59 (m, 2H), 3.35-3.27 (m, 2H), 2.93-2.83 (m, 1H), 2.62-2.51 (m, 2H), 2.46-2.38 (m, 2H), 2.35-2.25 (m, 4H), 2.05-1.98 (m, 1H), 1.78-1.71 (m, 2H), 1.49-1.41 (m, 4H), 1.37-1.31 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.1, 168.4, 166.9, 165.4, 162.9, 160.3, 159.4, 156.1, 140.6, 138.2, 137.6, 137.1, 133.3, 128.6 (2 x C), 127.6 (2 x C), 127.1 (2 x C), 121.5, 119.8, 119.0 (2 x C), 116.2, 115.2, 108.2, 68.8, 57.7, 53.0, 52.5, 48.8, 47.3, 41.6, 31.0, 28.4, 26.6, 26.2, 25.3, 22.1. ESI-HRMS m / z: Calcd for C 40 H 42 O6N7 + [M+ H] + 716.3191; found 716.3191.

[0210] Synthesis of compound C5: Compound 16c and compound 8a were reacted to give compound C5 in yield: 35%; 1 H NMR (500 MHz, DMSO-d6) δ 11.10 (s, 1H), 9.86 (s, 1H), 8.61 (d, J = 5.0 Hz, 1H), 8.27-8.22 (m, 2H), 7.97-7.93 (m, 2H), 7.80 (dd, J = 8.5, 7.0 Hz, 1H), 7.68-7.63 (m, 4H), 7.58-7.54 (m, 2H), 7.50 (d, J = 8.5 Hz, 1H), 7.47 (d, J = 5.0 Hz, 1H), 7.46-7.42 (m, 3H), 7.33-7.29 (m, 1H), 5.08 (dd, J = 13.0, 5.5 Hz, 1H), 4.20 (t, J = 6.5 Hz, 2H), 3.68-3.36 (m, 4H), 2.93-2.83 (m, 1H), 2.62-2.51 (m, 2H), 2.48-2.39 (m, 2H), 2.38-2.28 (m, 4H), 2.05-1.99 (m, 1H), 1.81-1.73 (m, 2H), 1.54-1.42 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.1, 168.3, 166.9, 165.4, 162.9, 160.2, 159.4, 156.0, 140.1, 140.0, 138.2, 137.6, 137.1, 133.3, 133.1, 128.9 (2 x C), 127.6 (2 x C), 127.1 (2 x C), 126.8 (2 x C), 126.7, 126.1 (2 x C), 119.8, 119.2 (2 x C), 116.2, 115.2, 108.3, 68.8, 57.6, 53.1, 52.5, 48.8, 47.2, 41.6, 31.0, 28.3, 25.8, 23.3, 22.0. ESI-HRMS m / z: calcd for C 45 H 44 O6N7 + [M + H] + 778.3348; found 778.3342.

[0211] Synthesis of compound C6: Compound 16c and compound 8b were reacted to give compound C6 in yield: 37%; 1 H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.89 (s, 1H), 8.61 (d, J = 5.2 Hz, 1H), 8.27-8.23 (m, 2H), 7.97-7.93 (m, 2H), 7.80 (dd, J = 8.4, 7.2 Hz, 1H), 7.68-7.63 (m, 4H), 7.58-7.54 (m, 2H), 7.50 (d, J = 8.4 Hz, 1H), 7.47 (d, J = 5.2 Hz, 1H), 7.46-7.41 (m, 3H), 7.33-7.28 (m, 1H), 5.07 (dd, J = 12.8, 5.2 Hz, 1H), 4.18 (t, J = 6.4 Hz, 2H), 3.70-3.54 (m, 2H), 3.36-3.29 (m, 2H), 2.93-2.81 (m, 1H), 2.62-2.51 (m, 2H), 2.46-2.37 (m, 2H), 2.36-2.24 (m, 4H), 2.05-1.97 (m, 1H), 1.79-1.69 (m, 2H), 1.50-1.40 (m, 4H), 1.38-1.29 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.1, 168.3, 166.9, 165.4, 162.9, 160.2, 159.4, 156.1, 140.1, 140.0, 138.2, 137.6, 137.1, 133.3, 133.1, 128.9 (2 x C), 127.6 (2 x C), 127.1 (2 x C), 126.8 (2 x C), 126.7, 126.1 (2 x C), 119.8, 119.2 (2 x C), 116.2, 115.2, 108.3, 68.8, 57.7, 53.3, 52.5, 48.8, 47.2, 41.7, 31.0, 28.4, 26.6, 26.2, 25.3, 22.1. ESI-HRMS m / z: calcd for C 46 H 46 O6N7 +[M + H] + Found 792.3504; Calculated 792.3504.

[0212] Synthesis of compound C7: Compound 16d and compound 8a were reacted to give compound C7 in yield of 36%; 1 H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.63 (s, 1H), 8.55 (d, J = 5.2 Hz, 1H), 8.25 - 8.18 (m, 2H), 7.80 (dd, J = 8.4, 7.2 Hz, 1H), 7.73 - 7.69 (m, 2H), 7.56 - 7.52 (m, 2H), 7.51 (d, J = 8.4 Hz, 1H), 7.44 (d, J = 7.2 Hz, 1H), 7.41 (d, J = 5.2 Hz, 1H), 7.18 - 7.13 (m, 2H), 5.08 (dd, J = 12.8, 5.2 Hz, 1H), 4.20 (t, J = 6.4 Hz, 2H), 3.68 - 3.57 (m, 2H), 3.34 - 3.29 (m, 2H), 2.92 - 2.82 (m, 1H), 2.62 - 2.52 (m, 2H), 2.47 - 2.39 (m, 3H), 2.37 - 2.27 (m, 4H), 2.04 - 1.98 (m, 1H), 1.78 (d, J = 8.8 Hz, 6H), 1.72 - 1.66 (m, 1H), 1.54 - 1.44 (m, 4H), 1.41 - 1.32 (m, 4H), 1.29 - 1.23 (m, 1H). 13C NMR (101 MHz, DMSO-d6) δ 172.9, 170.1, 168.4, 166.9, 165.4, 162.8, 160.3, 159.3, 156.1, 140.9, 138.3, 138.1, 137.7, 137.1, 133.3, 127.5 (2 x C), 127.0 (2 x C), 126.7 (2 x C), 119.8, 119.1 (2 x C), 116.2, 115.2, 107.9, 68.8, 57.6, 53.1, 52.5, 48.8, 47.3, 43.2, 41.8, 34.2 (2 x C), 31.0, 28.3, 26.5 (2 x C), 25.8, 25.7, 23.3, 22.1. ESI-HRMS m / z: calcd for C 45 H 50 O6N7 + [M+ H] + 784.3817; found 784.3813.

[0213] Synthesis of compound C8: Compound 16d and compound 8b were reacted to give compound C8 in yield of 38%; 1H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 9.63 (s, 1H), 8.55 (d, J = 5.2 Hz, 1H), 8.23–8.19 (m, 2H), 7.80 (dd, J = 8.4, 7.2 Hz, 1H), 7.73–7.70 (m, 2H), 7.56–7.53 (m, 2H), 7.51 (d, J = 8.4 Hz, 1H), 7.44 (d, J = 7.2 Hz, 1H), 7.41 (d, J = 5.2 Hz, 1H), 7.17–7.14 (m, 2H), 5.08 (dd, J = 12.8, 5.2 Hz, 1H), 4.19 (t, J = 6.4 Hz, 2H), 3.65–3.59 (m, 2H), 3.34–3.27 (m, 2H), 2.93–2.82 (m, 1H), 2.62–2.51 (m, 2H), 2.46–2.38 (m, 3H), 2.34–2.26 (m, 4H), 2.05–1.99 (m, 1H), 1.78 (d, J = 9.6 Hz, 6H), 1.71–1.67 (m, 1H), 1.48–1.42 (m, 4H), 1.39–1.32 (m, 6H), 1.29–1.24 (s, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 172.9, 170.0, 168.3, 166.9, 165.4, 162.8, 160.3, 159.3, 156.1, 140.9, 138.3, 138.1, 137.7, 137.1, 133.3, 127.5 (2 x C), 127.0 (2 x C), 126.7 (2 x C), 119.8, 119.1 (2 x C), 116.2, 115.2, 107.9, 68.8, 57.6, 53.0. 52.5, 48.8, 47.3, 43.2, 41.6, 34.2 (2 x C), 31.0, 28.4, 26.6, 26.5 (2 x C), 26.2, 25.7, 25.3, 22.0. ESI-HRMS m / z: Calcd for C 46 H 52 O6N7 + [M + H] + , 798.3974; Found, 798.3968.

[0214] Performance test

[0215] 1. Evaluation of the in vitro JAK1 degradation effect of the compound

[0216] Experimental method:

[0217] (1) Raw264.7 macrophages were selected and cultured in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO2.

[0218] (2) Drug treatment: After overnight culture and adhesion, different compounds (3 μM) were pre-administered for 1 h, and LPS (1 μg / mL) was stimulated for 6 h.

[0219] (3) JAK1 protein degradation detection: After 6 h of LPS (1 μg / mL) induction, Western blot was used to detect the change in JAK1 protein level, and the results are shown in Figure 1 .

[0220] Figure 1 The results are shown in FIG. 1, which are the screening results of the JAK1 protein degradation activity of each compound (A) and the heat map of the JAK1 protein degradation rate (B). In B, the color represents the degradation rate range, in which blue represents a lower degree of degradation, and red represents a higher degree of degradation. Figure 1 The results show that compounds A8, C2, C4 and C6 have better degradation effect on JAK1 protein, and therefore these four compounds are selected for subsequent activity detection and screening.

[0221] 2. Cell toxicity detection of the compound

[0222] Experimental method:

[0223] (1) Raw264.7 macrophages were selected and cultured in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO2.

[0224] (2) Drug treatment: After overnight culture and adhesion, different compounds (0.47-60 μM) were administered for 24 h.

[0225] (3) Cell toxicity detection: After 24 h of administration, CCK-8 method was used to detect the cell activity, the absorbance was measured at 450 nm, and the cell survival rate was calculated, and the results are shown in Figure 2 .

[0226] Figure 2 The results of the cell toxicity experiment of compounds A8, C2, C4 and C6 are shown in FIG. 2, expressed as a percentage of the control group, and the data are expressed as mean ± standard deviation (n=3), ***P<0.001 compared with the control group. Figure 2Results showed that compounds A8, C2 and C4 had no significant toxicity to RAW264.7 cells (survival rate > 90%) in the experimental concentration range.

[0227] 3. Compound DC 50 detection

[0228] Experimental method:

[0229] (1) Raw264.7 macrophages were selected and placed in DMEM medium containing 10% fetal bovine serum and cultured at 37℃, 5% CO2.

[0230] (2) Drug treatment: After overnight culture and adhesion of cell inoculation, different compounds (0.47-60 μM) were pre-administered for 1 h, and LPS (1 μg / mL) was stimulated for 6 h.

[0231] (3) JAK1 protein degradation detection: After LPS (1 μg / mL) induction for 6 h, Western blot was used to detect the change of JAK1 protein level, and the results are shown in Figure 3 .

[0232] Figure 3 Screening results (left) and degradation rate curve (right) of JAK1 protein degradation activity in RAW264.7 cells after treatment with compounds A8 (A), C2 (B) and C4 (C). Figure 3 Results showed that compounds A8, C2 and C4 could dose-dependently reduce the level of JAK1, and the degradation of JAK1 by A8, C2 and C4 showed a dose-dependent relationship, and their DC 50 were 1.40 μM, 4.0 μM and 0.85 μM, respectively.

[0233] 4. Detection of in vitro anti-inflammatory activity of compounds:

[0234] Experimental method:

[0235] (1) Raw264.7 macrophages were selected and placed in DMEM medium containing 10% fetal bovine serum and cultured at 37℃, 5% CO2.

[0236] (2) Drug treatment: After overnight culture and adhesion of cell inoculation, different compounds (0.47-60 μM) were pre-administered for 1 h, and LPS (1 μg / mL) was stimulated for 6 h.

[0237] (3) Inflammatory factor detection: After LPS (1 μg / mL) induction for 24 h, the levels of NO (Griess method) and IL-6, TNF-α (ELISA method) were detected, and the results are shown in Figure 4 .

[0238] Figure 4A8, C2 and C4 on LPS-induced RAW264.7 cells, wherein A is NO production, B is IL-6 level, and C is TNF-a level; the cell treatment concentration range is 0.05-60 μM, the treatment time is 24 h, the data are expressed as mean ± standard deviation (n = 3), *P < 0.05, ***P < 0.001 compared with the LPS group. Figure 4 The results show that compounds A8, C2 and C4 have dose-dependent inhibitory effect on NO production, and A8 can significantly reduce NO production at a concentration as low as 0.469 μM. ELISA results further show that A8 has the strongest anti-inflammatory effect, and its IC 50 values for IL-6 and TNF-a are 12.89 μM and 17.17 μM, respectively; the IC 50 values of C2 and C4 for IL-6 are 18.51 μM and 33.44 μM, respectively, and the IC 50 values for TNF-a are greater than 60 μM and 21.05 μM, respectively.

[0239] 5. Selective detection of compound A8 on JAK isozyme degradation:

[0240] Experimental method:

[0241] (1) Raw264.7 macrophages were selected and cultured in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO2.

[0242] (2) Drug treatment: after overnight culture of cell inoculation and adhesion, different compounds were pre-administered for 1 h, and LPS (1 μg / mL) was stimulated.

[0243] (3) JAKs protein degradation detection: after 6 h of LPS (1 μg / mL) induction, Western blot was used to detect the protein level changes of JAK2, JAK3 and TYK2, and the results are shown in Figure 5 .

[0244] Figure 5 Screening results (left) and degradation rate curve (right) of compound A8 on JAK2 (A), JAK3 (B) and TYK2 (C) degradation activity. Figure 5 The results show that A8 has a dose-dependent degradation effect on JAK2, and the DC 50 value is 0.92 μM, while it has no significant effect on JAK3 and TYK2 (the DC 50 values are both greater than 60 μM), suggesting that it has strong selectivity for JAK1 / 2 subtypes.

[0245] 6. Animal efficacy evaluation

[0246] Objective: To evaluate the anti-inflammatory effect of compound A8 in DSS-induced acute colitis mouse model.

[0247] Animal grouping: C57BL / 6J mice were randomly divided into blank control group, model group, A8 low-dose group (10 mg / kg), medium-dose group (20 mg / kg), high-dose group (40 mg / kg), and positive control group (5-aminosalicylic acid, 5-ASA).

[0248] Modeling and drug administration: Except for the blank group, the rest of the mice were allowed to drink 2.5% DSS solution freely for 7 days, while daily gavage administration was performed.

[0249] General indicators: Body weight was monitored, disease activity index (DAI) was recorded, colon length and spleen index were recorded.

[0250] Histology and inflammation analysis: H&E and Ab-PAS staining were performed on the colon to evaluate the mucosal structure, goblet cell number and mucus secretion; IHC was used to detect MPO and iNOS expression; ELISA was used to detect serum and colon tissue IL-6, IL-1β, TNF-α, IL-10 levels, and the results are shown in Figure 6-8 .

[0251] Figure 6 Results of the improvement of compound A8 on the symptoms of DSS-induced colitis mice, where A is the animal experiment design diagram, B is the body weight change during the experiment, C is the disease activity index (DAI) score, D is the colon length, E is the spleen index, F is the representative picture of the colon of each group, G is the representative picture of the spleen of each group, the values are expressed as mean ± standard deviation (n=8), compared with the model group, *P<0.05, **P<0.01, ***P<0.001. Figure 6 The results showed that in the DSS-induced colitis model group, the mice showed significant weight loss, increased disease activity index (DAI) score, significantly shortened colon length, and increased spleen index accompanied by splenomegaly, indicating that the acute colitis model was successfully constructed. Compared with the model group, the mice in the A8 treatment group showed different degrees of improvement in various indicators, including maintaining body weight, reducing DAI score, restoring colon length, and reducing spleen index. Notably, the medium and high dose A8 (20 mg / kg and 40 mg / kg) groups had more significant effects, and were comparable or even superior to the 5-ASA group (positive control).

[0252] Figure 7The results of intestinal histopathological evaluation of DSS-induced colitis mice, in which A is the images of colon H&E staining and Alcian blue-PAS staining, B is the results of immunohistochemical staining of colon tight junction proteins ZO-1 and Occludin, scale bar = 200 μm. The results of H&E staining show that there are significant pathological changes in the colon tissues of DSS-treated mice, including epithelial exfoliation, crypt damage and a large number of inflammatory cell infiltration. After A8 treatment, these pathological injuries are alleviated to varying degrees, showing that the mucosal structure is relatively complete and the inflammatory infiltration is reduced. To further evaluate the mucosal barrier function, Alcian blue-PAS (Ab-PAS) staining was used to visualize goblet cells and mucus secretion, and the results showed that the number of goblet cells in the model group was significantly reduced, and the mucus staining was weakened, suggesting that the intestinal barrier function was impaired; while the A8 treatment group can effectively restore the number of goblet cells and improve mucus secretion. IHC results show that the brown positive staining of tight junction proteins ZO-1 and Occludin in the colon tissues of the model group is significantly weakened and unevenly distributed, suggesting that DSS-induced colitis leads to impaired tight junction structure and decreased barrier function. After A8 treatment, the expression of ZO-1 and Occludin is enhanced and shows a relatively continuous distribution, indicating that A8 can restore the expression of tight junction proteins to some extent and improve the integrity of the intestinal barrier.

[0253] Figure 8 The results of inflammatory factor levels, blood routine and immunohistochemical analysis of DSS-induced colitis mice, in which A is the levels of IL-6, IL-10, IL-1β and TNF-α in serum (ELISA detection), B is the levels of IL-6, IL-10, IL-1β and TNF-α in colon tissue (ELISA detection), C is the analysis of peripheral blood immune cell subtypes, including total white blood cells (WBCs), lymphocytes, monocytes and neutrophils, the dashed line represents the upper and lower limits of the normal range, D is the immunohistochemical staining of MPO and iNOS in colon tissue, the positive signal is brown, scale bar = 200 μm, the numerical value is represented by mean ± standard deviation (n = 8), compared with the model group, *P < 0.05, **P < 0.01, ***P < 0.001. ELISA results show that the levels of IL-6, IL-10, IL-1β and TNF-α in the serum (A) and colon tissues (B) of DSS-induced mice are significantly increased, indicating that DSS-induced colitis can cause systemic inflammation and intestinal inflammation. Figure 8 Figure 8 ​The levels of pro-inflammatory cytokines IL-6, IL-1β and TNF-α were significantly increased, while the level of anti-inflammatory cytokine IL-10 was significantly decreased in B) of the above, suggesting that strong systemic and local inflammatory responses occurred. Compared with the model group, the A8 treatment group showed varying degrees of anti-inflammatory effects, mainly manifested as decreased levels of pro-inflammatory cytokines and restored IL-10 expression. Peripheral immune cell analysis showed that the number of peripheral blood white blood cells (WBC), lymphocytes, monocytes and neutrophils in the model group was significantly increased, suggesting that DSS-induced colon damage triggered systemic immune activation. After A8 treatment, the number of the above cells decreased significantly, suggesting that it had effective systemic anti-inflammatory effects. Myeloperoxidase (MPO) is a commonly used marker of neutrophil infiltration and acute inflammation; inducible nitric oxide synthase (iNOS) is expressed by activated macrophages and epithelial cells during the inflammatory process, which can cause tissue damage. Immunohistochemical results showed that the expression of MPO and iNOS in the colon tissue of DSS-treated mice was significantly enhanced, suggesting that the inflammatory activity was increased; while the A8 treatment group could significantly reduce the expression of these two markers, suggesting that it could effectively relieve the inflammatory response.

[0254] The above, only for the preferred specific embodiments of the present application, but the protection scope of the present application is not limited to this, any person skilled in the art in the technical range disclosed by the present application, can easily think of changes or replacement, should be covered within the scope of protection of the present application.

Claims

1. A PROTAC degrader, characterized in that Including the compound represented by formula (1) or its pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, cocrystal, deuterated product, Formula (1); Where L is 、 or ; R is H, 、 or ; G1 and G2 are independently a bond, CH2 or O, and G1 and G2 are not simultaneously a bond; n and m are independently 1, 2, 3 or 4.

2. The PROTAC degrader according to claim 1, characterized in that The L is 、 、 、 、 、 、 、 or .

3. The PROTAC degrader according to claim 1, characterized in that The structural formula of the PROTAC degrader is 、 、 or .

4. The PROTAC degrader according to claim 3, characterized in that The structural formula of the PROTAC degrader is: 。 5. A method for preparing a PROTAC degrader according to any one of claims 1 to 4, characterized in that: The following steps are involved: (1) Preparation of the compound of formula VI: ； a) mixing the compound of formula I with N,N-dimethylformamide dimethyl acetal and reacting at 50-120° C. for 10-14 hours to obtain a compound of formula II; b) mixing the compound of formula III with cyanamide and hydrochloric acid and reacting at 50-120° C. for 8-12 hours to obtain a compound of formula IV; c) mixing the compound of formula II obtained in step a) with the compound of formula IV obtained in step b) and reacting them at 80-120° C. for 40-60 h to obtain a compound of formula V; d) hydrolyzing the compound of formula V obtained in step c) at 20-80° C. for 1-8 hours to obtain a compound of formula VI; (2) Preparation of the compound of formula XI: ； e) reacting the compound of formula VII and the compound of formula VIII in the presence of N,N-diisopropylethylamine at 50-100° C. for 2-8 hours to obtain a compound of formula IX; f) reacting the compound of formula IX obtained in step e) with an HCl-dioxane solution at 20-50° C. for 1-3 h to obtain a compound of formula X; g) reacting the compound of formula X obtained in step f) with the compound of formula VI obtained in step d) at 20-50° C. for 8-12 h to obtain a compound of formula XI; Or (3) Preparation of the compound of formula XV: ； h) mixing the compound of formula X obtained in step f) with the compound of formula XII and reacting the mixture at 20-50° C. for 8-12 hours to obtain a compound of formula XIII; i) mixing the compound of formula XIII obtained in step h) with an HCl-dioxane solution and reacting the mixture at 20-50° C. for 1-3 hours to obtain a compound of formula XIV; j) mixing the compound of formula XIV obtained in step i) with the compound of formula VI obtained in step d) and reacting the mixture at 20-50° C. for 8-12 hours to obtain a compound of formula XV; Or (4) Preparation of the compound of formula XXI: ； k) mixing the compound of formula VI obtained in step d) with the compound of formula XVI and reacting them at 20-50° C. for 10-14 hours to obtain a compound of formula XVII; i) mixing the compound of formula XVII obtained in step k) with an HCl-dioxane solution and reacting the mixture at 20-50° C. for 1-3 hours to obtain a compound of formula XVIII; m) reacting the compound of formula XIX in the presence of a Dess-Martin periodinane at 20-50° C. for 2-4 h to obtain a compound of formula XX; n) reacting the compound of formula XX obtained in step m) and the compound of formula XVIII obtained in step i) with glacial acetic acid and sodium triacetoxyborohydride at 20-50° C. for 14-18 hours to obtain a compound of formula XXI; In Formula VIII, Formula IX, Formula X, Formula XI, Formula XIII, Formula XIV, Formula XV, Formula XIX, Formula XX and Formula XXI, R is H, 、 or ; G1 and G2 are independently a bond, CH2 or O, and G1 and G2 are not simultaneously a bond; n and m are independently 1, 2, 3 or 4.

6. A pharmaceutical composition, characterized in that Comprising the PROTAC degrader according to any one of claims 1 to 4 and a pharmaceutically acceptable excipient or excipient.

7. Use of the PROTAC degrader according to any one of claims 1 to 4 or the pharmaceutical composition according to claim 6 in the preparation of a medicament for preventing and / or treating inflammatory diseases, respiratory diseases, skin diseases, immune system diseases or colitis.

8. The use according to claim 7, characterized in that The inflammatory disease is an inflammatory skin disease; the respiratory disease is chronic obstructive pulmonary disease, colitis or asthma; the skin disease is psoriasis or atopic dermatitis; and the immune system disease is systemic lupus erythematosus or rheumatoid arthritis.

9. Use of the PROTAC degrader according to any one of claims 1 to 4 or the pharmaceutical composition according to claim 6 in the preparation of a medicament or JAK inhibitor for preventing and / or treating a JAK-mediated disease.

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