5, 7-dihydroxyflavone derivative, preparation method and application thereof, and CSF-1R inhibitor

Abstract

The invention relates to the technical field of medicine synthesis, in particular to a 5, 7-dihydroxy flavone derivative, a preparation method and application thereof and a CSF-1R inhibitor. The 3, 5, 7-dihydroxyflavone derivative is selected from any one of compounds shown in the following structural formula: # imgabs0 # imgabs1 #, and the 3, 5, 7-dihydroxyflavone derivative can be effectively combined with CSF-1R, so that the 3, 5, 7-dihydroxyflavone derivative has a good treatment effect on cancers.

Description

Technical Field

[0001] The present invention relates to the technical field of drug synthesis, in particular to a 5,7-dihydroxyflavone derivative, a preparation method and application thereof, and a CSF-1R inhibitor. Background Art

[0002] As a natural product, chrysin exhibits significant pharmacological activity in anti-tumor, antioxidant, and anti-inflammatory settings. Existing studies have demonstrated that chrysin not only effectively regulates immune cell function but also inhibits tumor cell proliferation and induces apoptosis, thus holding broad application prospects in tumor immunotherapy. Despite this, research on the molecular mechanisms of action of chrysin in the tumor immune microenvironment remains relatively scarce, particularly with regard to the identification and validation of its specific immune targets. Therefore, elucidating the potential immune targets of chrysin and exploring its detailed mechanisms of action are of great theoretical significance for promoting its application in tumor immunotherapy.

[0003] Colony-stimulating factor 1 receptor (CSF1R, also known as fms) is a receptor tyrosine kinase (RTK) encoded by the c-Fms proto-oncogene and a type I single-transmembrane protein primarily expressed on the surface of monocytes and macrophages. The CSF1 / CSF1R signaling pathway plays a key regulatory role in macrophage proliferation and differentiation in solid tumors and is a key factor in macrophage phenotypic changes. Notably, CSF1R activation in the tumor microenvironment (TME) is closely associated with tumor metastasis and growth, as well as poor prognosis. CSF1R is often abnormally overexpressed in breast cancer, ovarian cancer, Hodgkin's lymphoma, skin, and peripheral T-cell lymphomas, and is activated by CSF1 in an autocrine or paracrine-dependent manner. Binding to CSF1R regulates macrophage proliferation, survival, and differentiation, recruiting macrophages to the TME to promote their transformation into an immunosuppressive phenotype (i.e., M2 macrophages). These M2 macrophages promote tumor growth, metastasis, and angiogenesis by secreting a variety of immunosuppressive factors (such as IL-10 and TGF-β) and factors that promote tumor angiogenesis (such as VEGF), thereby inhibiting anti-tumor immune responses. This not only enhances the tumor-promoting function of TAMs, but also directly promotes the release of inflammatory mediators from tumor cells to trigger an inflammatory cascade, driving the continued progression of tumors. The CSF1 / CSF1R signaling pathway modulates the survival of TAMs to change the cellular microenvironment to maintain the immunosuppressive state in the TME, and the presence of CSF1R+ macrophages in tumors is closely associated with poor prognosis in various tumor types. In the TME, CSF1R plays a key role by regulating the macrophage population TAMs. As the core mechanism for regulating TAM function, the CSF1 / CSF1R signaling pathway is a key target for the development of new anti-tumor therapies and has important research and application value. Therefore, targeting the CSF1 / CSF1R signaling pathway has become an effective strategy to eliminate TAMs, reduce the recruitment of M2 macrophages, or reshape their immune phenotype.

[0004] The application of CSF-1R in the tumor microenvironment still faces several challenges. Currently, research on the role of CSF-1R in specific cell types (such as endothelial cells, dendritic cells and regulatory T cells) is still insufficient, and the existing research results are inconsistent, which makes it difficult to draw clear conclusions. Therefore, the specific functions and molecular mechanisms of CSF-1R in these cells have not been fully elucidated, which to some extent limits its development in targeted therapy strategies. The same problem is that although CSF-1R is highly expressed in macrophages, its expression in cancer cells is low, which makes it difficult to effectively detect in some experiments. Natural products have significant advantages in drug discovery due to their unique active skeletons, active groups and excellent biological activities. However, no natural products have been found to significantly inhibit CSF-1R activity.

[0005] In summary, there are still many directions and issues that need to be studied and explored regarding CSF-1R. Future research should focus on in-depth exploration of CSF-1R signaling in different cell types and its potential role in tumor immunity. At the same time, exploring the effects of natural products on CSF-1R and studying combined treatment strategies will help improve the efficacy of CSF-1R targeted therapy and promote its widespread implementation in clinical applications.

[0006] In view of this, the present invention is proposed. Summary of the Invention

[0007] The present invention aims to provide a 5,7-dihydroxyflavone derivative, a preparation method thereof, its application and a CSF-1R inhibitor. The present invention provides a novel compound that can effectively bind to CSF-1R and thereby have a good therapeutic effect on cancer.

[0008] The present invention is achieved in that:

[0009] In a first aspect, the present invention provides a 5,7-dihydroxyflavone derivative selected from any one of the compounds represented by the following structural formulas:

[0010]

[0011] Wherein, R is selected from Any one of the functional groups formed;

[0012] R' is selected from C1-C10 substituted or unsubstituted alkyl, R" is selected from hydrogen or C1-C5 unsubstituted alkyl, represents a 4-7 membered heterocycloalkyl group containing one N, R' represents a C1-C8 unsubstituted alkyl group, R"" represents a C1-C3 unsubstituted alkyl group, represents a 4- to 7-membered heterocycloalkyl group containing 2 N groups, where n is any value between 1 and 5;

[0013] R1 is selected from substituted phenyl or substituted thiophene, and m is any value between 1-5.

[0014] In a second aspect, an embodiment of the present invention provides a method for preparing a 7-dihydroxyflavone derivative, comprising: selecting any one of the following synthetic pathways for synthesis:

[0015] Path 1:

[0016]

[0017] Path 2:

[0018]

[0019] In a third aspect, the present invention provides a CSF-1R inhibitor comprising the 5,7-dihydroxyflavone derivative described in the aforementioned embodiment.

[0020] In a fourth aspect, the present invention provides a use of a 7-dihydroxyflavone derivative in the preparation of an anti-tumor drug.

[0021] The present invention has the following beneficial effects: the embodiment of the present invention modifies the structure of chrysin and then provides a new 5,7-dihydroxyflavone derivative, which has a good inhibitory effect on CSF-1R kinase and a good therapeutic effect on tumors such as colon cancer and breast cancer, and can be used in the treatment of tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0023] Figure 1 This is the kinase spectrum of the 5,7-dihydroxyflavone derivative 5c provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0024] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0025] In a first aspect, an embodiment of the present invention provides a 5,7-dihydroxyflavone derivative selected from any one of the compounds represented by the following formula 1 and formula 2:

[0026] Wherein, R is selected from Any one of the functional groups formed.

[0027] R' is selected from C1-C10 substituted or unsubstituted alkyl groups, for example, preferably C1-C6 unsubstituted alkyl groups or C3-C6 cycloalkyl substituted C1-C4 alkyl groups; or more preferably C1-C4 unsubstituted alkyl groups or C3-C6 cycloalkyl substituted C1-C4 alkyl groups. For example, it includes but is not limited to any one of methyl, ethyl, isopropyl, n-propyl, isobutyl, n-butyl, tert-butyl, n-pentyl, n-hexyl, and cyclopropyl substituted ethyl groups.

[0028] R" is selected from hydrogen or C1-C5 unsubstituted alkyl.

[0029] represents a 4- to 7-membered heterocycloalkyl group containing one N, such as azetidine, azetidine, azetidine and azetidine.

[0030] R'' represents a C1-C8 unsubstituted alkyl group. R"" represents a C1-C3 unsubstituted alkyl group.

[0031] represents a 4- to 7-membered heterocycloalkyl group containing 2 N atoms, for example, tetrahydropyrazine.

[0032] n is any value between 1 and 5, for example, 1, 2, 3, 4, 5, or any other value between 1 and 5.

[0033] It should be noted that the above Indicates the position of bonding to the parent nucleus.

[0034] m is any value between 1 and 5, for example, 1, 2, 3, 4, 5, or any other value between 1 and 5.

[0035] R1 is selected from substituted phenyl or substituted thiophene. The substituted phenyl is monosubstituted phenyl or trisubstituted phenyl, and the substituent of the monosubstituted phenyl is located at the para position. The substituent of the substituted phenyl is selected from any one of halogen, nitro, cyano, C1-C5 unsubstituted alkyl, C1-C3 aldehyde, and C1-C3 amide.

[0036] The substituted thiophene is a disubstituted thiophene; preferably, the substituent of the substituted thiophene is a halogen.

[0037] It should be noted that the halogens provided in the embodiments of the present invention include but are not limited to chlorine, bromine and iodine.

[0038] The C1-C10 unsubstituted alkyl group, C1-C8 unsubstituted alkyl group, C1-C6 unsubstituted alkyl group and C1-C5 unsubstituted alkyl group provided in the embodiments of the present invention include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, tert-pentyl and other alkyl groups.

[0039] The C3-C6 cycloalkyl groups provided in the embodiments of the present invention include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and other C3-C6 cycloalkyl groups.

[0040] The C1-C3 aldehyde groups provided in the embodiments of the present invention include, but are not limited to, formaldehyde, acetaldehyde and n-propionaldehyde.

[0041] The C1-C3 amide groups provided in the embodiments of the present invention include but are not limited to formamide and acetamide.

[0042] Specifically, it is selected from any one of the compounds represented by the following structural formulas:

[0043]

[0044] In a second aspect, an embodiment of the present invention provides a method for preparing a 5,7-dihydroxyflavone derivative, comprising: selecting any one of the following synthesis pathways for synthesis:

[0045] Path 1:

[0046]

[0047] Specifically,

[0048] Step a: Compound 1, Compound 2 and an acid-binding agent (for example, including but not limited to potassium carbonate) are mixed to carry out a substitution reaction, wherein the reaction conditions include: 40-80° C., 4-8 h;

[0049] Step b: subjecting compound 3 to ester hydrolysis reaction under alkaline conditions (such as but not limited to sodium hydroxide); the reaction conditions include: 40-80° C., 4-8 h;

[0050] Step c: reacting compound 4, a coupling agent (e.g., TBTU), an acid-binding agent (e.g., DIPEA), and an R-containing amine to form an amide bond; the reaction conditions include: room temperature (20-30° C.), 10-14 h;

[0051] Then hydrolysis is carried out, and the reaction conditions include: 40-80°C, 4-8h.

[0052] The compounds provided in the examples of the present invention are used for illustration, with reference to the following synthesis pathway:

[0053]

[0054] The overall reaction process is as follows: Commercially available chrysin (compound 1) undergoes a substitution reaction with ethyl bromoacetate (compound 2) to obtain compound 3, which is then hydrolyzed under alkaline conditions to produce the key compound 4. Compound 4 then reacts with different amines under the catalysis of coupling agents O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU) and N,N-diisopropylethylamine (DIPEA) to form amide bonds to obtain some target products 5a-m. Finally, 5a-m is hydrolyzed under alkaline conditions to produce the target products 6a-j.

[0055] Among them, experimental reagents and reaction conditions: (a) K2CO3, DMSO, 60℃, 6h, yield: 80-90%; (b) NaOH, H2O, 60℃, 6h, yield: 80-90%; (c) TBTU, DIPEA, anhydrous DMF, rt, 12h, yield: 50-60%; (d) NaOH, H2O, 60℃, 6h, yield: 80-90%.

[0056] Path 2:

[0057]

[0058] Wherein, step d: Compound 7a-k is mixed with compound 8 to undergo substitution reaction to form a sulfonamide bond; the reaction conditions include: 40-80° C., 4-8 h;

[0059] Step e: Compounds 9a-k, a condensing agent, compound 1 and a catalyst are mixed and reacted. The reaction conditions include: reflux for 20-30 hours.

[0060] The overall reaction process is as follows: Commercially available compounds 7a-k undergo a substitution reaction with alanine (compound 8) to form a sulfonamide bond to obtain intermediates 9a-k. Then, under the action of a condensing agent 1,3-dicyclohexylcarbodiimide (DCC) and a catalyst 4-dimethylaminopyridine (DMAP), compounds 9a-k react with compound 1 to form an ester bond to obtain the final product 10a-k.

[0061] Specifically, experimental reagents and reaction conditions: (d) NaOH, H2O, 65°C, 6h, yield: 60-90%; (e) DCC, DMAP, DCM, rt, 24h, yield: 40-50%.

[0062] After each step of the reaction, post-treatment is required to obtain a compound with higher purity. The post-treatment methods include pH adjustment, rotary evaporation, drying, and extraction, etc., which will not be described in detail in the embodiments of the present invention.

[0063] In a third aspect, the present invention provides a CSF-1R inhibitor comprising the 5,7-dihydroxyflavone derivative described in the aforementioned embodiment.

[0064] In a fourth aspect, the present invention provides a use of a 7-dihydroxyflavone derivative in the preparation of an anti-tumor drug.

[0065] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0066] Examples 1-23

[0067] Examples 1-23 provide a method for synthesizing 5,7-dihydroxyflavone derivatives (denoted as 5a-m and 6a-j), respectively, with reference to the following synthetic routes:

[0068]

[0069] Experimental reagents and reaction conditions: (a) K2CO3, DMSO, 60℃, 6h, yield: 80-90%; (b) NaOH, H2O, 60℃, 6h, yield: 80-90%; (c) TBTU, DIPEA, anhydrous DMF, rt, 12h, yield: 50-60%; (d) NaOH, H2O, 60℃, 6h, yield: 80-90%.

[0070] The specific steps are as follows:

[0071] 1. Step a

[0072] Chrysin (Compound 1) (2.54 g, 0.01 mol, 1 equivalent) and anhydrous potassium carbonate (2.76 g, 0.02 mol, 2 equivalents) were added to 100 mL of dimethyl sulfoxide (DMSO) solution and stirred. Ethyl bromoacetate (2.00 g, 0.012 mol, 1.2 equivalents) was then added dropwise to the solution. The reaction mixture was then stirred at 60°C for 6 hours. TLC confirmed the completion of the reaction. After the reaction solution cooled, dilute sulfuric acid was added dropwise to adjust the pH to 1-2. A yellow solid precipitated, which was filtered to obtain a precipitate. Subsequently, the precipitate was recrystallized by heating it in an isopropanol solution until the precipitate was completely dissolved. The solution was then cooled to room temperature. During the process, light yellow crystals precipitated and were filtered to obtain compound 3, which was used in the next reaction with a yield of approximately 80-90%.

[0073] 2. Step b

[0074] Compound 3 (3.40 g, 0.01 mol, 1 equivalent) was added to a 2 mol / L aqueous sodium hydroxide solution (150 mL) and stirred at 60°C for 6 hours until the solution became clear. The reaction was complete as determined by TLC. After the reaction solution cooled, dilute hydrochloric acid was added dropwise to adjust the pH to 2-3. A large amount of white solid precipitated, which was filtered, washed three times with ultrapure water, and dried to obtain compound 4 as a white powder, which was used directly in the next reaction with a yield of approximately 80-90%.

[0075] 3. Step c

[0076] Compound 4 (3.12 g, 0.01 mol, 1 equivalent) and different amines (0.011 mol, 1.1 equivalents) were added to 100 mL of anhydrous N,N-dimethylformamide (DMF) solution, followed by the addition of O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU) (3.85 g, 0.012 mol, 1.2 equivalents) and N,N-diisopropylethylamine (DIPEA) (1.94 g, 0.015 mol, 1.5 equivalents) to the solution and stirred at room temperature for 12 h. TLC was used to monitor the reaction until complete. Ultrapure water (25 mL) was then added to quench the reaction. Solids precipitated during the reaction and were filtered to obtain the precipitate. The precipitate was washed three times with saturated NaHCO3 solution and then dried. The resulting residue was dry-loaded onto a 12 g flash preparative liquid chromatography column and purified by silica gel chromatography (Pure C-810 produced by BUCHI, eluent: dichloromethane:methanol = 100:1) to obtain the target products 5a-m in yields of approximately 50-60%.

[0077] 4. Step d

[0078] A portion of the target product 5a-m (5 mmol, 1 equivalent) was added to a 2 mol / L aqueous sodium hydroxide solution (150 mL) and stirred at 60°C for 6 hours until the solution became clear. The reaction was complete as determined by TLC. After the reaction solution cooled, dilute hydrochloric acid was added dropwise to adjust the pH to 2-3. A large amount of white solid precipitated, which was filtered, washed three times with ultrapure water, and dried to obtain the target product 6a-j in an approximately 80-90% yield.

[0079] Examples 24-35

[0080] Examples 24-35 provide a method for synthesizing 5,7-dihydroxyflavone derivatives (denoted as 10a-k), respectively, with reference to the following synthetic routes:

[0081]

[0082] Experimental reagents and reaction conditions: (d) NaOH, H2O, 65°C, 6 h, yield: 60-90%; (e) DCC, DMAP, DCM, rt, 24 h, yield: 40-50%.

[0083] The specific steps are as follows:

[0084] 1. Step d

[0085] Alanine (compound 8) (445 mg, 5 mmol, 1 equivalent) was dissolved in a 2 mol / L aqueous sodium hydroxide solution. Another starting material, compound 7a-k (5 mmol, 1 equivalent), was added over 30 minutes and stirred at room temperature for 6 hours. After TLC analysis, the reaction was completed. Dilute hydrochloric acid was added to adjust the pH to 1-2, and a large amount of white solid precipitated. White compounds 9a-k were obtained by filtration and used directly in the next reaction with a yield of approximately 60-90%.

[0086] 2. Step e

[0087] 1,3-Dicyclohexylcarbodiimide (DCC) (2.27 g, 0.011 mol, 1.1 eq) and 4-dimethylaminopyridine (DMAP) (24 mg, 2 mmol, 0.2 eq) were dissolved in anhydrous dichloromethane (100 mL). Compound 1 (2.54 g, 0.01 mol, 1 eq) and intermediate 9a-k (0.01 mol, 1 eq) were then added at 0°C. The mixed reaction solution was stirred at room temperature for 24 hours. After TLC was performed to determine the reaction completion, the filtrate was filtered to remove the large amount of insoluble byproducts produced during the reaction. Anhydrous sodium sulfate was then added to the filtrate for drying. The filtrate was then evaporated under reduced pressure using a rotary evaporator. The resulting residue was dry-loaded onto a 12 g flash preparative liquid chromatography column and purified by silica gel chromatography (Pure C-810 manufactured by BUCHI, eluent: dichloromethane:methanol = 100:1) to obtain the target products 10a-k in approximately 40-50% yields.

[0088] Characterization

[0089] The characterization data of the 5,7-dihydroxyflavone derivatives prepared in Examples 1-35 are as follows:

[0090] Example 1: Ethyl (2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7yl)oxy)acetyl)glycine ester (Compound 5a).

[0091]

[0092] White powder, yield is about 73%. 1H NMR (400MHz, DMSO-d6, ppm) δ12.80(s,1H),8.63(s,1H),8.08(d,J=7.4Hz,2H),7.60(dd,J=11.8,7.1Hz,3H),7.03( s,1H),6.82(s,1H),6.44(s,1H),4.72(s,2H),4.11(q,J=7.1Hz,2H),3.93(d,J=5.9Hz,2H),1.19(t,J=7.1Hz,3H).

[0093] 13 C NMR(101MHz,DMSO-d6,ppm)δ182.56,170.04,168.00,163.99,161.55,157.63,132.65 ,130.99,129.60,126.89,105.87,105.79,99.35,94.07,67.48,61.02,40.99,14.52.

[0094] HRMS(ESI):calcd.C 21 H 19 NO7,[M+Na] + m / z:420.1054; found:420.1055.

[0095] Example 2: Ethyl 3-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)propanoate (Compound 5b).

[0096]

[0097] White powder, yield is about 47%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.80 (s, 1H), 8.25 (t, J = 5.7Hz, 1H), 8.16-7.88 (m, 2H), 7.66-7.42 (m, 3H), 7.03 (s, 1H), 6.78 ( d,J=2.3Hz,1H),6.41(d,J=2.3Hz,1H),4.62(s,2H),4.04(q,J=7.1Hz,2H),3.40(t,2H),2.51(t,2H),1.16(t,J=7.1Hz,3H).

[0098] 13C NMR (101MHz, DMSO-d6, ppm) δ182.54,171.68,167.33,164.04,164.00,161.56,157.63,132.6 3,130.99,129.60,126.89,105.86,105.74,99.26,93.97,67.58,60.43,35.06,34.13,14.49.

[0099] HRMS(ESI):calcd.C 22 H 21 NO7,[M+Na] + m / z:434.1210; found:434.1216.

[0100] Example 3: Ethyl 3-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetylamino)butanoate (Compound 5c).

[0101]

[0102] Yellow powder, yield about 85%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.79(s,1H),8.12(s,1H),8.06(d,J=7.5Hz,2H),7.59(dq,J=14.5,7.1Hz,3H),7.01(s,1H),6.75(s,1H ), 6.40 (s, 1H), 4.60 (s, 2H), 4.25 (p, J = 7.0Hz, 1H), 4.02 (q, J = 7.1Hz, 2H), 2.51 (dtd, J = 21.9, 15.2, 6.9Hz, 2H), 1.14 (t, J = 6.8Hz, 6H).

[0103] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.52,171.15,166.40,164.11,163.97,161.56,157.60,132.6 1,130.98,129.58,126.86,105.83,105.70,99.24,93.99,67.61,60.33,42.22,20.56,14.48.

[0104] HRMS(ESI):calcd.C 23 H 23 NO7,[M+Na] + m / z:448.1367; found:448.1369.

[0105] Example 4: Ethyl 4-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetylamino)butanoate (Compound 5d).

[0106]

[0107] White powder, yield is about 89%. 1 H NMR(400MHz,DMSO-d6,ppm)δ12.79(s,1H),8.22(t,J=5.8Hz,1H),8.07(dt,J=6.8,1.6Hz,2H),7.76-7.47(m,3H),7.03(s,1H),6.79(s,1H), 6.43 (s, 1H), 4.63 (s, 2H), 4.01 (q, J = 7.1Hz, 2H), 3.17 (q, J = 6.6Hz, 2H), 2.29 (t, J = 7.5Hz, 2H), 1.70 (p, J = 7.1Hz, 2H), 1.14 (t, J = 7.1Hz, 3H).

[0108] 13 C NMR(101MHz,DMSO-d6,ppm)δ182.54,173.06,167.27,164.09,163.99,161.56,157.63,132.62,1 30.99,129.60,126.87,105.85,105.71,99.30,93.97,67.66,60.22,38.12,31.31,24.89,14.53.

[0109] HRMS(ESI):calcd.C 23 H 23 NO7,[M+Na]+m / z:448.1367; found:448.1377.

[0110] Example 5: Methyl 5-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)pentaciate (Compound 5e).

[0111]

[0112] Yellow powder, yield is about 82%. 1H NMR (400MHz, DMSO-d6, ppm) δ12.79 (s, 1H), 8.19 (t, J = 5.9Hz, 1H), 8.10-7.94 (m, 2H), 7.81-7.42 (m, 3H), 7.01 (s, 1H), 6.77(s,1H),6.41(s,1H),4.62(s,2H),3.55(s,3H),3.15(q,J=6.3Hz,2H),2.29(t,J=7.0Hz,2H),1.68-1.26(m,4H).

[0113] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.52,173.66,167.11,164.08,163.95,161.56,157.61,132.59,1 30.98,129.57,126.85,105.82,105.70,99.27,93.95,67.67,51.61,38.38,33.32,28.91,22.25.

[0114] HRMS(ESI):calcd.C 23 H 23 NO 7, [M+Na] + m / z:448.1367; found:448.1371.

[0115] Example 6: Methyl 6-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)hexanoate (Compound 5f).

[0116]

[0117] Yellow powder, yield about 47%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.79 (s, 1H), 8.16 (t, J = 5.8Hz, 1H), 8.12-7.98 (m, 2H), 7.68-7.50 (m, 3H), 7.02 (s, 1H), 6.78 (s, 1H), 6.42 (s, 1H), 4.62 (s, 2H), 3.55 (s, 3H), 3.13 (q, J = 6.6Hz, 2H), 2.24 (t, J = 7.4Hz, 2H), 1.47 (dp, J = 22.3, 7.3Hz, 4H), 1.24 (qd, J = 9.8, 9.1, 5.9Hz, 2H).

[0118] 13C NMR (101MHz, DMSO-d6, ppm) δ182.52,173.71,167.05,164.11,163.96,161.56,157.60,132.60,130. 98,129.58,126.85,105.84,105.69,99.28,93.98,67.68,51.61,38.58,33.62,29.16,26.24,24.58.

[0119] HRMS(ESI):calcd.C 24 H 25 NO7,[M+Na] + m / z:462.1523; found:462.1524.

[0120] Example 7: Ethyl 7-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)heptanoate (Compound 5g).

[0121]

[0122] Pale yellow powder, yield is about 43%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.79 (s, 1H), 8.15 (t, J = 5.8Hz, 1H), 8.11-8.03 (m,2H),7.59(dq,J=14.2,7.0Hz,3H),7.02(s,1H),6.78(s,1H),6.42(s,1H) ,4.62(s,2H),4.01(q,J=7.1Hz,2H),3.13(q,J=6.6Hz,2H),2.20(t,J=7.4Hz ,2H),1.44(dp,J=14.4,7.5Hz,4H),1.28-1.18(m,4H),1.15(t,J=7.1Hz,3H).

[0123] 13 C NMR(101MHz,DMSO-d6,ppm)δ182.53,173.25,167.03,164.13,163.96,161.57,157.61,132.61,130.98, 129.59,126.86,105.85,99.28,93.98,67.70,60.08,38.72,33.87,29.34,28.62,26.47,24.85,14.56.

[0124] HRMS(ESI):calcd.C 26 H29 NO7,[M+Na] + m / z:490.1836; found:490.1842.

[0125] Example 8: Ethyl 1-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetyl)piperidine-4-carboxylate (Compound 5h).

[0126]

[0127] Yellow powder, yield is about 64%. 1 H NMR(400MHz,DMSO-d6,ppm)δ12.80(s,1H),8.17-7.90(m,2H),7.68-7.46(m,4H),7.02(s,1H),6 .78(s,1H),6.40(s,1H),5.09-4.95(m,2H),4.19(dd,J=10.3,6.4Hz,1H),4.08(q,J=7.1Hz,2H), 3.78(d,J=13.6Hz,1H),3.20-3.09(m,1H),2.85-2.75(m,1H),2.63(tt,J=11.1,4.0Hz,1H),1.87 (dt,J=13.2,8.8Hz,2H),1.71-1.57(m,1H),1.42(qd,J=11.6,4.1Hz,1H),1.19(t,J=7.1Hz,3H).

[0128] 13 C NMR(101MHz,DMSO-d6,ppm)δ182.51,174.31,165.24,164.74,163.94,161.55,157.60,132.59,1 31.04,129.59,126.89,105.81,105.49,99.09,94.11,66.56,60.47,43.72,28.58,28.04,14.54.

[0129] HRMS(ESI):calcd.C 25 H 25 NO7,[M+Na] + m / z:474.1523; found:474.1524.

[0130] Example 9: Methyl 1-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetyl)pyrrolidine-3-carboxylate (Compound 5i).

[0131]

[0132] Yellow powder, yield is about 64%. 1 H NMR(400MHz,DMSO-d6,ppm)δ12.79(d,J=1.5Hz,1H),8.19-7.94(m,2H),7.72-7.46(m,3H),7.00(s,1H),6.76(dd,J=9.7,2.3Hz,1H),6 .39(dd,J=3.5,2.2Hz,1H),4.94-4.79(m,2H),3.83-3.56(m,5H),3.56-3.44(m,1H),3.23(dp,J=47.6,7.5Hz,1H),2.28-1.91(m,2H).

[0133] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.49,173.67,173.34,165.35,165.30,164.68,163.91,161.54,132.57,131. 02,129.56,126.86,105.78,105.50,99.10,99.07,94.06,94.04,52.39,52.36,43.25,41.06,29.30,27.15.

[0134] HRMS(ESI):calcd.C 23 H 21 NO7,[M+Na] + m / z:446.1210; found:446.1208.

[0135] Example 10: Ethyl 3-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetylamino)-2,2-dimethylpropanoate (Compound 5j).

[0136]

[0137] Yellow powder, yield about 54%. 1H NMR (400MHz, DMSO-d6, ppm) δ12.79(s,1H),8.10-8.05(m,2H),8.01(t,J=6.4Hz,1H),7.67-7.52(m,3H),7.02(s,1H),6.75(d,J= 2.3Hz, 1H), 6.40 (d, J = 2.2Hz, 1H), 4.69 (s, 2H), 4.04 (q, J = 7.1Hz, 2H), 3.32 (d, J = 6.4Hz, 2H), 1.16 (t, J = 7.1Hz, 3H), 1.10 (s, 6H).

[0138] 13 C NMR(101MHz,DMSO-d6,ppm)δ182.52,176.15,167.55,164.18,163.97,161.61,157.61,132.63,1 30.99,129.59,126.86,105.84,105.68,99.13,93.97,67.49,60.67,46.39,43.44,23.28,14.41.

[0139] HRMS(ESI):calcd.C 24 H 25 NO7,[M+Na] + m / z:462.1523; found:462.1527.

[0140] Example 11: 5-Hydroxy-7-(2-(4-(2-hydroxyethyl)piperazin-1-yl)-2-oxoethoxy)-2-phenyl-4H-chroman-4-one (Compound 5k).

[0141]

[0142] Light yellow powder, yield is about 65%. 1 H NMR(400MHz,DMSO-d6,ppm)δ12.81(d,J=7.2Hz,1H),8.06(d,J=7.4Hz,2H),7 .58(dq,J=8.0,4.8,3.4Hz,3H),7.08-6.80(m,1H),6.75(d,J=2.2Hz,1H),6. 55-6.28(m,1H),5.76(d,J=1.6Hz,1H),4.98(d,J=8.6Hz,2H),4.30(t,J=5.6 Hz, 1H), 3.59-3.21 (m, 4H), 2.78-2.34 (m, 5H), 1.11 (dt, J = 85.7, 7.4Hz, 1H).

[0143] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.54,182.51,168.52,165.31,164.65,164.05,164.02,161.68,161.55,157.6 2,157.59,132.62,129.58,126.89,105.83,105.51,98.96,94.11,94.08,66.47,65.44,62.55,55.39,38.71.

[0144] HRMS(ESI):calcd.C 23 H 24 N2O6,[M+Na] + m / z:447.1526; found:447.1527.

[0145] Example 12: 1-((2-((5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)methyl)cyclopropane-1-carboxylate (Compound 51).

[0146]

[0147] Pale yellow powder, yield is about 87%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.78(s,1H),8.20-7.90(m,3H),7.58(dq,J=14.8,7.3Hz,3H),6.99(s,1H),6.72(s,1H),6.38( s,1H),4.67(s,2H),4.03(q,J=7.1Hz,2H),3.43(s,2H),1.15(t,J=7.1Hz,3H),1.05(t,J=3.5Hz,2H),0.94(t,J=3.6Hz,2H).

[0148] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.52,173.90,167.44,164.12,163.97,161.63,157.59,132.62,130.9 8,129.59,126.86,126.83,105.84,105.69,99.13,93.95,67.52,60.78,41.11,24.01,14.44,13.99.

[0149] HRMS(ESI):calcd.C 24 H23 NO7,[M+Na] + m / z:460.1367; found:460.1372.

[0150] Example 13: 1-((2-((5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)methyl)cyclopropane-1-carboxylate (Compound 5m).

[0151]

[0152] Pale yellow powder, yield is about 87%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.78(s,1H),8.20-7.90(m,3H),7.58(dq,J=14.8,7.3Hz,3H),6.99(s,1H),6.72(s,1H),6.38( s,1H),4.67(s,2H),4.03(q,J=7.1Hz,2H),3.43(s,2H),1.15(t,J=7.1Hz,3H),1.05(t,J=3.5Hz,2H),0.94(t,J=3.6Hz,2H).

[0153] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.52,173.90,167.44,164.12,163.97,161.63,157.59,132.62,130.9 8,129.59,126.86,126.83,105.84,105.69,99.13,93.95,67.52,60.78,41.11,24.01,14.44,13.99.

[0154] HRMS(ESI):calcd.C 24 H 23 NO7,[M+Na] + m / z:460.1367; found:460.1372.

[0155] Example 14: (2-((5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetyl)glycine (Compound 6a).

[0156]

[0157] Light yellow powder, yield is about 52%. 1H NMR (400MHz, DMSO-d6, ppm) δ12.80 (s, 2H), 8.53 (t, J = 6.0Hz, 1H), 8.27-7.95 (m, 2H), 7.71-7.43 ( m, 3H), 7.03 (d, J = 1.4Hz, 1H), 6.82 (dd, J = 6.7, 2.2Hz, 1H), 6.42 (s, 1H), 4.78 (s, 2H), 3.86 (s, 2H).

[0158] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.56,171.46,167.81,164.02,164.00,161.55,157.65,132.64,13 0.99,129.62,129.60,126.91,105.87,105.78,99.37,99.01,94.08,93.91,67.50,65.32,40.92.

[0159] HRMS(ESI):calcd.C 19 H 15 NO7,[M+Na] + m / z:392.0741; found:392.0738.

[0160] Example 15: 3-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)propanoic acid (Compound 6b).

[0161]

[0162] White powder, yield is about 56%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.81(s,1H),12.29(s,1H),8.24(t,J=5.7Hz,1H),8.17-7.92(m,2H),7.60(qd,J=8.6 ,7.7,3.6Hz,3H),7.05(s,1H),6.81(s,1H),6.44(s,1H),4.63(s,2H),3.67(t,J=5.0Hz,2H),2.46(t,J=7.0Hz,2H).

[0163] 13C NMR (101MHz, DMSO-d6, ppm) δ182.59,173.32,167.27,164.08,164.05,161.56,157.68 ,132.67,131.02,129.64,126.94,105.89,105.76,99.28,94.03,67.59,35.10,34.18.

[0164] HRMS(ESI):calcd.C 20 H 17 NO7,[M+Na] + m / z:406.0897; found:406.0898.

[0165] Example 16: 3-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetylamino)butanoic acid (Compound 6c).

[0166]

[0167] White powder, yield is about 78%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.80(s,1H),12.33(s,1H),8.12(s,1H),8.07(dt,J=6.8,1.6Hz,2H),7.65-7.49(m,3H),7.02(s,1H) ,6.79(s,1H),6.42(s,1H),4.60(s,2H),4.29-4.09(m,1H),2.57-2.46(m,1H),2.39(dd,J=15.6,7.3Hz,1H),1.15(d,J=6.6Hz,3H).

[0168] 13 C NMR(101MHz,DMSO-d6,ppm)δ182.54,172.86,166.32,164.10,163.99,161.57,157.62 ,132.62,131.00,129.60,126.89,105.84,105.71,99.22,94.06,67.62,42.18,20.57.

[0169] HRMS(ESI):calcd.C 21 H 19 NO7,[M+Na] + m / z:420.1054; found:420.1055.

[0170] Example 17: 4-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetylamino)butanoic acid (Compound 6d).

[0171]

[0172] White powder, yield is about 75%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.81 (s, 1H), 12.08 (s, 1H), 8.22 (t, J = 5.8Hz, 1H), 8.09 (d, 2H), 7.73-7.37 (m, 3H), 7.0 5(s,1H),6.82(s,1H),6.45(s,1H),4.64(s,2H),3.17(q,J=6.6Hz,2H),2.23(t,J=7.4Hz,2H),1.68(p,J=7.2Hz,2H).

[0173] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.59,174.69,167.22,164.14,164.04,161.56,157.67,13 2.66,131.03,129.64,126.93,105.90,105.74,99.33,94.02,67.66,38.26,31.44,24.94.

[0174] HRMS(ESI):calcd.C 21 H 19 NO7,[M+Na] + m / z:420.1054; found:420.1060.

[0175] Example 18: 5-(2-((5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)pentanoic acid (Compound 6e).

[0176]

[0177] White powder, yield is about 68%. 1H NMR (400MHz, DMSO-d6, ppm) δ12.80 (s, 1H), 12.09 (s, 1H), 8.22 (t, J = 5.8Hz, 1H), 8.11-7.99 (m, 2H), 7.67-7.52 (m, 3H) ,7.03(s,1H),6.80(s,1H),6.43(s,1H),4.63(s,2H),3.15(q,J=6.3Hz,2H),2.22(t,J=6.9Hz,2H),1.59-1.37(m,4H).

[0178] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.55,174.84,167.07,164.13,164.00,161.55,157.64,132.6 3,131.01,129.62,126.90,105.87,105.71,99.28,94.00,67.67,38.49,33.74,29.02,22.34.

[0179] HRMS(ESI):calcd.C 22 H 21 NO7,[M+Na] + m / z:434.1210; found:434.1211.

[0180] Example 19: 6-(2-((5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetamido)hexanoic acid (Compound 6f).

[0181]

[0182] Yellow powder, yield is about 52%. 1 H NMR(400MHz,DMSO-d6,ppm)δ12.79(s,1H),12.06(s,1H),8.18(t,J=5.8Hz,1H),8.11-7.99(m,2H),7.59(dq,J=14.2,6.9Hz,3H),7.01(s,1H), 6.78(s,1H),6.42(s,1H),4.61(s,2H),3.14(q,J=6.6Hz,2H),2.17(t,J =7.3Hz, 2H), 1.47 (dp, J = 14.7, 7.3Hz, 4H), 1.26 (td, J = 8.6, 4.0Hz, 2H).

[0183] 13C NMR(101MHz,DMSO-d6,ppm)δ182.52,174.91,167.04,164.10,163.97,161.55,157.60,132.61,1 30.98,129.59,126.86,105.84,105.69,99.26,94.00,67.67,38.66,34.03,29.25,26.36,24.66.

[0184] HRMS(ESI):calcd.C 23 H 23 NO7,[M+Na] + m / z:448.1367; found:448.1371.

[0185] Example 20: 7-(2-((5-hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetylamino)heptanoic acid (Compound 6g).

[0186]

[0187] Yellow powder, yield about 51%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.81(s,1H),12.04(s,1H),8.17(t,J=5.8Hz,1H),8.14-8.06(m,2H),7.60(qd,J=8.7,7.7,2.4Hz,3H),7.05( s,1H),6.81(s,1H),6.44(s,1H),4.63(s,2H),3.13(q,J=6.6Hz,2H),2.15(t,J=7.3Hz,2H),1.44(h,J=7.1,6.5Hz,4H),1.32-1.18(m,4H).

[0188] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.58,174.92,167.03,164.17,164.03,161.56,157.65,132.67,131. 02,129.64,126.92,105.90,105.72,99.31,94.05,67.71,38.76,34.04,29.38,28.74,26.55,24.89.

[0189] HRMS(ESI):calcd.C 24 H 25 NO7,[M+Na] +m / z:462.1523; found:462.1527.

[0190] Example 21: 1-(2-((5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetyl)piperidine-4-carboxylic acid (Compound 6h).

[0191]

[0192] Yellow powder, yield about 77%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.79(s,2H),8.06(t,J=6.9Hz,2H),7.59(dq,J=14.8,7.3Hz,3H ),7.01(d,J=5.7Hz,1H),6.78(d,J=11.9Hz,1H),6.39(s,1H),5.15-4.90(m,2H),4.85(s,2H) ,4.19(d,J=13.0Hz,1H),3.77(d,J=13.7Hz,1H),3.14(t,J=12.4Hz,1H),2.80(t,J=12.1Hz,1 H), 2.56 (dd, J = 10.1, 4.1Hz, 1H), 1.87 (t, J = 14.8Hz, 1H), 1.52 (dq, J = 81.8, 12.0, 11.3Hz, 1H).

[0193] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.53,176.04,169.96,164.23,163.96,161.60,157.65,13 2.60,129.58,126.90,105.78,105.65,98.99,93.89,65.31,43.85,41.13,28.66,28.11.

[0194] HRMS(ESI):calcd.C 23 H 21 NO7,[M+Na] + m / z:446.1210; found:446.1215.

[0195] Example 22: 1-(2-((5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetyl)pyrrolidine-3-carboxylic acid (Compound 6i).

[0196]

[0197] Yellow powder, yield about 70%.1 H NMR (400MHz, DMSO-d6, ppm) δ12.79(s,1H),8.19-7.95(m,2H),7.69-7.47(m,3H),7.00(d,J=5.9Hz,1H),6.85-6.58(m,1 H),6.39(dt,J=4.2,1.9Hz,1H),5.02-4.74(m,2H),3.81-3.44(m,4H),3.12(dp,J=47.7,7.5Hz,1H),2.26-1.91(m,2H).

[0198] 13 C NMR(101MHz,DMSO-d6,ppm)δ182.48,174.77,174.47,165.29,164.69,163.91,161.54,131.02,1 29.56,126.87,105.77,105.49,99.09,94.08,66.71,66.63,44.57,43.53,41.31,29.35,27.27.

[0199] HRMS(ESI):calcd.C 22 H 19 NO7,[M+Na] + m / z:432.1056; found:432.1054.

[0200] Example 23: 3-(2-((5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl)oxy)acetylamino)-2,2-dimethylpropanoic acid (Compound 6j).

[0201]

[0202] Light yellow powder, yield is about 57%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.72(s,1H),7.99(dd,J=7.2,5.0Hz,3H),7.55(dq,J=14.5,7.1Hz,3H),6. 89 (s, 1H), 6.68 (d, J = 2.2Hz, 1H), 6.33 (d, J = 2.2Hz, 1H), 4.64 (s, 2H), 3.29 (d, J = 6.3Hz, 2H), 1.06 (s, 6H).

[0203] 13C NMR(101MHz,DMSO-d6,ppm)δ182.43,178.40,167.76,164.03,163.98,161.49,157.53,13 2.64,130.80,129.59,126.78,105.63,105.61,99.07,93.86,67.35,46.19,43.05,23.32.

[0204] HRMS(ESI):calcd.C 22 H 21 NO7,[M+Na] + m / z:434.1210; found:434.1212.

[0205] Example 24: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-methylphenyl)sulfonamide)propanoate (Compound 10a).

[0206]

[0207] White powder, yield: 87%. 1 H NMR(400MHz,DMSO-d6,ppm)δ12.83(d,J=0.8Hz,1H),8.07(dt,J=8.6,1.7Hz,2 H),7.87(t,J=5.8Hz,1H),7.74(d,J=8.3Hz,2H),7.58(dtd,J=15.3,6.9,1.5Hz ,3H),7.42(d,J=8.1Hz,2H),7.13-7.08(m,1H),7.05(t,J=1.8Hz,1H),6.65(dd ,J=2.1,0.9Hz,1H),3.13(q,J=6.4Hz,2H),2.78(t,J=6.7Hz,2H),2.38(s,3H).

[0208] 13 C NMR (101MHz, DMSO-d6, ppm) δ183.01,169.35,164.61,161.19,156.65,156.15,143.27,137.77,132. 82,130.79,130.19,129.61,127.06,127.02,108.72,106.18,105.84,102.06,38.80,34.91,21.42.

[0209] HRMS(ESI):calcd.C 25 H 21NO7S,[M+Na] + m / z:502.0931; found:502.0936.

[0210] Example 25: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((2,4,6-trimethylphenyl)sulfonamide)propanoate (Compound 10b).

[0211]

[0212] Light green powder, yield: 62%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.83(s,1H),8.09(d,J=7.5Hz,2H),7.71(t,J=5.9Hz,1H),7.60(dt,J=14.0,6.7Hz,3H),7 .12(s,1H),7.04(s,3H),6.63(d,J=1.9Hz,1H),3.13(q,J=6.4Hz,2H),2.74(t,J=6.6Hz,2H),2.58(s,6H),2.25(s,3H).

[0213] 13 C NMR(101MHz,DMSO-d6,ppm)δ183.04,169.35,164.64,161.16,156.65,156.15,142.03,138.86,134.62, 132.85,132.17,130.82,129.65,127.05,108.71,106.21,105.81,102.02,34.70,33.83,22.99,20.83.

[0214] HRMS(ESI):calcd.C 27 H 25 NO7S,[M+Na] + m / z:530.1244; found:530.1249.

[0215] Example 26: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-ethylphenyl)sulfonamide)propanoate (Compound 10c).

[0216]

[0217] Pale yellow powder, yield: 57%. 1H NMR (400MHz, DMSO-d6, ppm) δ12.84 (s, 1H), 8.07 (d, J = 7.4Hz, 2H), 7.88 (t, J = 5 .8Hz,1H),7.76(d,J=8.0Hz,2H),7.59(dt,J=14.4,6.9Hz,3H),7.45(d,J=8.0H z,2H),7.11(s,1H),7.06(d,J=2.0Hz,1H),6.66(d,J=2.0Hz,1H),3.13(q,J=6 .4Hz, 2H), 2.78 (t, J = 6.6Hz, 2H), 2.68 (q, J = 7.7Hz, 2H), 1.19 (t, J = 7.5Hz, 3H).

[0218] 13 C NMR(101MHz,DMSO-d6,ppm)δ183.03,169.35,164.63,161.19,156.67,156.17,149.26,138.00,132.83, 130.80,129.63,129.05,127.16,127.03,108.73,106.19,105.86,102.08,38.82,34.93,28.46,15.56.

[0219] HRMS(ESI):calcd.C 27 H 25 NO6S,[M+H] + m / z:492.1476; found:492.1477.

[0220] Example 27: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-bromophenyl)sulfonamide)propanoate (Compound 10d).

[0221]

[0222] White powder, yield: 54%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.85 (s, 1H), 8.13-8.03 (m, 3H), 7.87-7.82 (m, 2H), 7.80-7.75 (m, 2H), 7.61 (ddd, J=14.4, 8.0, 6.1Hz, 3H), 7.14 (s, 1H), 7.06 (d, J=2.1Hz, 1H), 6.67 (d, J=2.0Hz, 1H), 3.16 (q, J=6.4Hz, 2H), 2.79 (t, J=6.6Hz, 2H).

[0223] 13 C NMR (101MHz, DMSO-d6, ppm) δ183.07, 169.29, 164.68, 161.20, 156.70, 156.14, 139.96, 132.86, 130. 83, 129.66, 129.59, 129.07, 127.08, 126.86, 126.84, 108.77, 106.24, 105.85, 102.08, 38.79.34.87.

[0224] HRMS(ESI):calcd.C 24 H 18 BrNO7S, [M+Na] + m / z: 565.9879; found: 565.9881.

[0225] Example 28: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-nitrophenyl)sulfonamide)propanoate (Compound 10e).

[0226]

[0227] Yellow powder, yield: 46%. 1 H NMR (400MHz, DMSO-d6, ppm) δ8.36 (d, J=8.6Hz, 2H), 8.01 (d, J=8.4Hz, 2H), 7.96 (d, J=7.4Hz, 2H), 7.51 (d, J=8.2Hz, 3H), 6.77 (s, 1H), 6.45 (s, 1H), 6.17 (s, 1H), 3.07 (d, J=7.4Hz, 2H), 2.41 (t, J=7.8Hz, 2H).

[0228] 13 C NMR (101MHz, DMSO-d6, ppm) δ182.19, 167.92, 164.61, 163.74, 161.75, 157.94, 153.58, 149.93, 146. 51, 131.96, 131.14, 129.17, 128.28, 128.25, 126.37, 124.48, 124.44, 105.22, 33.49, 31.86, 30.58.

[0229] HRMS(ESI):calcd.C 24 H 18 N2O9S, [M+H] + m / z: 511.0806; found: 511.0809.

[0230] Example 29: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-chlorophenyl)sulfonamide)propanoate (Compound 10f).

[0231]

[0232] White powder, yield: 56%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.85 (s, 1H), 8.10 (d, J = 7.6Hz, 2H), 8.07 (t, J = 5.6Hz, 1H), 7.85 (d, J = 8.4Hz, 2H), 7.70 (d, J = 8.4Hz , 2H), 7.65-7.57 (m, 3H), 7.15 (s, 1H), 7.07 (d, J=2.0Hz, 1H), 6.67 (d, J=2.0Hz, 1H), 3.16 (q, J=6.4Hz, 2H), 2.79 (t, J=6.6Hz, 2H).

[0233] 13 C NMR (101MHz, DMSO-d6, ppm) δ183.08, 169.30, 164.68, 161.20, 156.71, 156.14, 139.55, 137.89, 1 32.88, 130.84, 129.93, 129.67, 128.98, 127.09, 108.78, 106.25, 105.85, 102.09, 38.79, 34.87.

[0234] HRMS(ESI):calcd.C 24 H 18 ClNO7S, [M+Na] + m / z: 522.0385; found: 522.0390.

[0235] Example 30: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-fluorophenyl)sulfonamide)propanoate (Compound 10g).

[0236]

[0237] White powder, yield: 58%. 1H NMR (400MHz, DMSO-d6, ppm) δ12.85 (s, 1H), 8.12 (d, J=6.8Hz, 2H), 8.00 (t, J=5.8Hz, 1H), 7.91 (td, 2H), 7.66-7.57 (m, 3H), 7.50-7.44 (m, 2H), 7.16 (s, 1H), 7.09 (d, J=2.0Hz, 1H), 6.68 (d, J=2.0Hz, 1H), 3.14 (q, J=6.4Hz, 2H), 2.78 (t, J=6.7Hz, 2H).

[0238] 13 C NMR (101MHz, DMSO-d6, ppm) δ183.09, 169.33, 165.90, 164.70, 163.41, 161.20, 156.73, 156.17, 137.05, 137.02, 132.90, 130. 86, 130.10, 130.00, 129.97, 129.69, 127.10, 117.05, 116.97, 116.83, 116.75, 108.79, 106.26, 105.87, 102.13, 38.79, 34.87.

[0239] HRMS(ESI):calcd.C 24 H 18 FNO7S, [M+Na] + m / z: 506.0680; found: 506.0686.

[0240] Example 31: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-cyanophenyl)sulfonamide)propanoate (Compound 10h).

[0241]

[0242] Yellow crystals, yield: 76%. 1H NMR (400MHz, DMSO-d6, ppm) δ12.83 (s, 1H), 8.27 (d, J=1.8Hz, 1H), 8.21 (t, J=5.7Hz, 1H), 8.15 (d, J=7.5Hz, 2H), 8.07 (d, J=7.2Hz, 3H), 7.84 (t, J=7.9 Hz, 1H), 7.58 (ddd, J=14.5, 7.9, 6.2Hz, 3H), 7.10 (s, 1H), 7.05 (d, J=2.0Hz , 1H), 6.64 (d, J=2.0Hz, 1H), 3.21 (q, J=6.3Hz, 2H), 2.80 (t, J=6.5Hz, 2H).

[0243] 13 C NMR (101MHz, DMSO-d6, ppm) δ183.02, 169.29, 164.63, 161.21, 156.67, 156.10, 142.01, 136.61, 132.82, 131. 47, 131.29, 130.79, 130.62, 129.61, 127.03, 118.04, 113.02, 108.74, 106.19, 105.78, 102.02, 34.90, 33.82.

[0244] HRMS(ESI):calcd.C 25 H 18 N2O7S, [M+Na] + m / z: 513.0727; found: 513.0726.

[0245] Example 32: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((2,5-dichlorothiophene)-3-sulfonamide)propanoate (Compound 10i).

[0246]

[0247] Pale yellow powder, yield: 53%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.84 (s, 1H), 8.38 (t, J=5.6Hz, 1H), 8.11-8.05 (m, 2H), 7.59 (dt, J=14.4, 6.9Hz, 3H), 7 .36 (s, 1H), 7.11 (s, 1H), 7.07 (d, J=2.0Hz, 1H), 6.68 (d, J=2.0Hz, 1H), 3.32 (q, J=6.3Hz, 2H), 2.84 (t, J=6.6Hz, 2H).

[0248] 13 C NMR (101MHz, DMSO-d6, ppm) δ183.03, 169.24, 164.64, 161.22, 156.68, 156.13, 137.27, 132.83, 1 30.80, 129.63, 129.18, 127.19, 127.03, 126.85, 108.75, 106.21, 105.81, 102.04, 38.72, 34.85. HRMS(ESI):calcd.C 22 H 15 Cl2NO7S2, [M+Na] + m / z: 561.9559; found: 561.9561.

[0249] Example 33: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-acetylphenyl)sulfonamide)propanoate (Compound 10j).

[0250]

[0251] Pale yellow powder, yield: 47%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.82 (s, 1H), 8.17 (dd, J=12.6, 6.9Hz, 3H), 8.05 (d, J=7.7Hz, 2H), 7.98 (d, J=8.4Hz, 2H), 7. 57 (dt, J=14.7, 7.1Hz, 3H), 7.09 (s, 1H), 7.03 (d, J=1.9Hz, 1H), 6.63 (d, J=1.9Hz, 1H), 3.18 (t, J=6.3Hz, 2H), 2.62 (s, 3H).

[0252] 13 C NMR (101MHz, DMSO-d6, ppm) δ197.73, 182.99, 169.26, 164.60, 161.17, 156.63, 156.09, 144.39, 139.94, 132.80, 130.77, 129.60, 129.56, 127.36, 127.01, 108.71, 106.16, 105.78, 102.00, 38.81.34.92.27.45.

[0253] HRMS(ESI):calcd.C 26 H 21 NO8S, [M+Na] + m / z: 530.0880; found: 530.0879.

[0254] Example 34: 5-Hydroxy-4-oxo-2-phenyl-4H-chroman-7-yl 3-((4-acetamidophenyl)sulfonamide)propanoate (Compound 10k).

[0255]

[0256] White powder, yield: 38%. 1 H NMR (400MHz, DMSO-d6, ppm) δ12.82 (s, 1H), 8.17 (dd, J=12.6, 6.9Hz, 3H), 8.05 (d, J=7.7Hz, 2H), 7.98 (d, J=8.4Hz, 2H), 7. 57 (dt, J=14.7, 7.1Hz, 3H), 7.09 (s, 1H), 7.03 (d, J=1.9Hz, 1H), 6.63 (d, J=1.9Hz, 1H), 3.18 (t, J=6.3Hz, 2H), 2.62 (s, 3H).

[0257] 13 C NMR (101MHz, DMSO-d6, ppm) δ197.73, 182.99, 169.26, 164.60, 161.17, 156.63, 156.09, 144.39, 139.94, 132.80, 130.77, 129.60, 129.56, 127.36, 127.01, 108.71, 106.16, 105.78, 102.00, 38.81.34.92.27.45.

[0258] HRMS(ESI):calcd.C 26 H 22 N2O8S, [M+H] + m / z: 523.1170; found: 523.1173.

[0259] Experimental Example 1

[0260] The in vitro anti-tumor activity of the 5,7-dihydroxyflavone derivatives provided in Examples 1-34 of the present invention was detected using the Cell Counting Kit 8 (CCK8) method.

[0261] Among them, the cells used are as follows: six types of adherent cells, including A549 (human non-small cell lung cancer cells), HCT-116 (human colon cancer cells), U87 (human glioma cells), HGC-27 (human gastric cancer cells), MDB-MA-231 (human triple-negative breast cancer cells), MCF-7 (human breast cancer cells) and HUVEC (human umbilical vein endothelial cells).

[0262] The details are as follows: According to 1.5×10 5 cells·mL -1 A 96-well plate was inoculated with 100 μL of liquid per well. The edge of the plate was left uninoculated, and 100 μL of PBS buffer was added to the wells to prevent edge effects. After inoculation, the 96-well plate was transferred to an incubator and incubated for 24 hours before dosing.

[0263] The 5,7-dihydroxyflavone derivatives and chrysin provided in Examples 1-34 were set to five concentrations, specifically 10 μM, 5.0 μM, 2.50 μM, 1.25 μM, and 0.625 μM. 10 μL of the corresponding compound was then added to each well of the cell plate (three wells per compound). 10 μL of the corresponding complete medium was added to three wells of the blank control group, with three replicates, and a blank control group was set up. The administered compounds were then placed in an incubator and cultured for 48 hours.

[0264] Under a dark background, add 10 μL of CCK8 solution to each experimental well of the cell plate after 48 h of culture (avoid bubbles during the addition process to affect the experimental results), then place the cell plate in the incubator and culture for 2 h, and then place the cell plate in a microplate reader to detect the OD value.

[0265] The calculation formula for cell viability is:

[0266]

[0267] The results are shown in Table 1.

[0268] Table 1 In vitro anti-tumor cell proliferation activity

[0269]

[0270]

[0271]

[0272] a After all cells were treated with 10 μM of the compound for 72 hours, the average inhibition rate was determined using the CCK-8 assay. All data are derived from three independent experiments and are presented as the mean.

[0273] At the same time, the IC values of some 5,7-dihydroxyflavone derivatives were tested. 50 , the results are shown in Table 2.

[0274] Table 2 IC values of some 5,7-dihydroxyflavone derivatives 50

[0275]

[0276]

[0277] From Table 1 and Table 2, it can be seen that the compounds provided in the embodiments of the present invention can effectively inhibit tumor cells and have a therapeutic effect on tumors.

[0278] The HUVEC cytotoxicity test was used to evaluate the biosafety of some 5,7-dihydroxyflavone derivatives 5a, 5b, 5c, 5d, 5g, 5l, 6c, 6e, 6f, 10h, 10i, 10j and 10k. The results are shown in Table 3.

[0279] Table 3 Evaluation results of HUVEC cytotoxicity

[0280]

[0281] According to Table 3, the cytotoxicity of the 5,7-dihydroxyflavone derivatives provided in the embodiments of the present invention is within the safe range, which proves that the 5,7-dihydroxyflavone derivatives provided in the embodiments of the present invention have good biocompatibility.

[0282] Experimental Example 2

[0283] Testing the CSF-1R inhibitory activity of 5,7-dihydroxyflavone derivatives

[0284] The specific method is as follows:

[0285] The radioactive element-labeled kinase activity test method uses γ-32P-ATP or γ-33P-ATP as a substrate in the reaction to carry out the phosphorylation reaction. After the reaction is completed, the radioactive labeled product generated will be bound to the filter membrane, and the unreacted radioisotope will be washed away to ensure that only the phosphorylated product is accurately detected. In this way, the phosphorylation level can be accurately measured and the interference of unreacted substrates or radioisotopes can be effectively avoided. Therefore, in order to explore the effect of the synthesized inhibitor on kinase activity, this study selected Eurofins' KinaseProfiler TM The service conducts kinase inhibitory activity analysis, which can accurately evaluate the inhibitory effects of different inhibitors on target kinases and provide valuable data support for subsequent drug development and biological research.

[0286] Results see Figure 1 and Table 4.

[0287] Table 4 Kinase test results of 5,7-dihydroxyflavone derivatives on CSF-1R

[0288]

[0289] According to the above results, the 5,7-dihydroxyflavone derivatives provided by the embodiments of the present invention have a good inhibitory effect on CSF-1R kinase.

[0290] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. A 5,7-dihydroxyflavone derivative, characterized in that It is selected from any one of the compounds represented by the following structural formulas: Wherein, R is selected from Any one of the functional groups formed; R' is selected from C1-C10 substituted or unsubstituted alkyl, R" is selected from hydrogen or C1-C5 unsubstituted alkyl, represents a 4-7 membered heterocycloalkyl group containing one N, R' represents a C1-C8 unsubstituted alkyl group, R"" represents a C1-C3 unsubstituted alkyl group, represents a 4- to 7-membered heterocycloalkyl group containing 2 N groups, where n is any value between 1 and 5; R1 is selected from substituted phenyl or substituted thiophene, and m is any value between 1-5.

2. The 5,7-dihydroxyflavone derivative according to claim 1, characterized in that R′ is selected from C1-C6 unsubstituted alkyl or C3-C6 cycloalkyl substituted C1-C4 alkyl; Preferably, R' is selected from any one of methyl, ethyl, isopropyl, n-propyl, isobutyl, n-butyl, tert-butyl, n-pentyl, n-hexyl and cyclopropyl-substituted ethyl.

3. The 5,7-dihydroxyflavone derivative according to claim 1, characterized in that The substituted phenyl group in R1 is a monosubstituted phenyl group or a trisubstituted phenyl group; Preferably, the substituent of the monosubstituted phenyl group is located in the para position; Preferably, the substituent of the substituted phenyl group is selected from any one of halogen, nitro, cyano, C1-C5 unsubstituted alkyl, C1-C3 aldehyde and C1-C3 amide.

4. The 5,7-dihydroxyflavone derivative according to claim 1, characterized in that The substituted thiophene in R1 is a disubstituted thiophene; Preferably, the substituent of the substituted thiophene is halogen.

5. The 5,7-dihydroxyflavone derivative according to claim 1, characterized in that It is selected from any one of the compounds represented by the following structural formulas:

6. A method for preparing the 5,7-dihydroxyflavone derivative according to claim 1, characterized in that: include: Choose any of the following synthesis routes: Path 1: Path 2:

7. The preparation method according to claim 6, characterized in that include: Step a: Compound 1, Compound 2 and an acid-binding agent are mixed to carry out a substitution reaction, wherein the reaction conditions include: 40-80° C., 4-8 h; Step b: subjecting compound 3 to ester hydrolysis under alkaline conditions; the reaction conditions include: 40-80° C., 4-8 h; Step c: reacting compound 4, a coupling agent, an acid-binding agent, and an R-containing amine to form an amide bond; the reaction conditions include: room temperature, 10-14 hours; Then hydrolysis is carried out, and the reaction conditions include: 40-80°C, 4-8h.

8. The preparation method according to claim 6, characterized in that include: Step d: Compound 7a-k is mixed with compound 8 to undergo a substitution reaction to form a sulfonamide bond; The reaction conditions include: the reaction conditions include: 40-80°C, 4-8h; Step e: Compounds 9a-k, a condensing agent, compound 1 and a catalyst are mixed and reacted. The reaction conditions include: room temperature, 20-30 hours.

9. A CSF-1R inhibitor, characterized in that It comprises the 5,7-dihydroxyflavone derivative according to claim 1.

10. Use of the 5,7-dihydroxyflavone derivative according to claim 1 in the preparation of anti-tumor drugs.

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