Compounds containing benzothiazole or benzoxazole, preparation methods and applications thereof

Abstract

The present invention relates to the technical field of compound synthesis, and more specifically, to a compound containing benzothiazole or benzoxazole, a preparation method thereof, and an application thereof. The structural formula of the compound containing benzothiazole or benzoxazole is as follows: wherein Y represents O or S, X represents hydrogen or halogen, and R represents any one of a substituted or unsubstituted C3-C8 cycloalkyl, a substituted or unsubstituted phenyl, a substituted or unsubstituted C1-C10 alkyl, and a substituted or unsubstituted C3-C10 alkenyl. The compound can be used as a soluble epoxide hydrolase inhibitor, thereby having a certain therapeutic effect on diseases caused by soluble epoxide hydrolase disorders.

Description

Technical Field

[0001] The present invention relates to the technical field of compound synthesis, and in particular to a compound containing benzothiazole or benzoxazole, a preparation method thereof and an application thereof. Background Art

[0002] Soluble epoxide hydrolase (sEH) inhibitors are promising candidates for the treatment of diseases caused by sEH dysregulation, including the treatment or alleviation of renal deterioration, hypertension, vascular inflammation, obstructive pulmonary disease, interstitial lung disease, and asthma. However, current research focuses primarily on hypertension and the cardiovascular diseases it causes. Advances in understanding human sEH have facilitated the rational design of inhibitors. In particular, the innovative pharmacodynamic model for human sEH (hsEH) inhibitors proposed by Hammoc et al. has made the design and synthesis of hsEH inhibitors more rational and feasible. Subsequently, research on sEH inhibitors has progressed from initially exhibiting only micromolar inhibitory activity in vitro and minimal or no activity in vivo to currently exhibiting nanomolar activity in vitro and excellent pharmacokinetic properties in vivo. However, numerous challenges remain with sEH inhibitors. For example, 12-(3-adamantan-1-yl-ureido)-dodecanoic acid (AUDA) exhibits significant antihypertensive and anti-inflammatory effects in various animal models. However, their poor metabolic stability and limited water solubility make them difficult to use pharmacologically. With the exception of trans-4-[4-(4-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB), which has glioma inhibitory activity, no other compounds have been reported to exhibit tumor suppressive activity. Therefore, the development of more active compounds is urgently needed. Summary of the Invention

[0003] The present invention aims to provide a benzothiazole- or benzoxazole-containing compound, a preparation method thereof, and its application. The present invention provides a novel benzothiazole- or benzoxazole-containing compound that can act as a soluble epoxide hydrolase inhibitor, thereby having a certain therapeutic effect on diseases caused by soluble epoxide hydrolase disorders.

[0004] The present invention is achieved in that:

[0005] In a first aspect, the present invention provides a compound containing benzothiazole or benzoxazole, the structural formula of which is shown below:

[0006] Wherein, Y represents O or S, X represents hydrogen or halogen, and R represents any one of substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted C1-C10 alkyl and substituted or unsubstituted C3-C10 alkenyl.

[0007] In an optional embodiment, when Y is O, X represents halogen, and R represents any one of C2-C6 unsubstituted alkyl, C4-C7 unsubstituted cycloalkyl, substituted and unsubstituted phenyl;

[0008] Preferably, halogen represents chlorine or bromine;

[0009] Preferably, the substituted phenyl group is a monosubstituted phenyl group; more preferably, it is a para-substituent.

[0010] In an optional embodiment, when Y is O and X represents chlorine, R represents a C2-C6 unsubstituted linear alkyl group, a phenyl group, a C6-C7 unsubstituted cycloalkyl group, or a monosubstituted phenyl group, and the substituent of the monosubstituted phenyl group is selected from a C1-C5 carbonyl group;

[0011] Preferably, when Y is O and X represents chlorine, R represents a C3-C6 unsubstituted straight-chain alkyl, a phenyl, an unsubstituted cyclohexane, or a para-monosubstituted phenyl, and the substituent of the para-monosubstituted phenyl is selected from a C1-C3 carbonyl group.

[0012] In an alternative embodiment, when Y is O and X represents bromine, R represents a C2-C6 unsubstituted linear alkyl group, a C5-C7 unsubstituted cycloalkyl group, or a monosubstituted phenyl group, and the substituent of the monosubstituted phenyl group is selected from a C1-C5 ether group;

[0013] Preferably, when Y is O and X represents bromine, R represents a C3-C6 unsubstituted linear alkyl group, a C5-C7 unsubstituted cycloalkyl group, or a para-substituted phenyl group, and the substituent of the para-substituted phenyl group is selected from a C1-C3 ether group.

[0014] In an alternative embodiment, when Y is S, X represents bromine and chlorine, and R represents C1-C10 unsubstituted alkyl and monosubstituted phenyl;

[0015] Preferably, R represents a C1-C8 unsubstituted straight-chain alkyl group and a para-monosubstituted phenyl group, and the substituent is selected from a C1-C5 unsubstituted straight-chain alkyl group.

[0016] In an optional embodiment, when Y is S and X represents bromine, R represents a monosubstituted phenyl group; the substituent of the monosubstituted phenyl group is selected from C1-C10 unsubstituted alkyl groups; preferably C1-C8 unsubstituted linear alkyl groups; more preferably C5-C8 unsubstituted linear alkyl groups.

[0017] In an optional embodiment, when Y is S and X represents chlorine, R represents a para-monosubstituted phenyl group, and the substituent of the para-monosubstituted phenyl group is selected from C1-C5 unsubstituted straight-chain alkyl groups; preferably C1-C3 unsubstituted straight-chain alkyl groups.

[0018] In an optional embodiment, the compound containing benzothiazole or benzoxazole is selected from any one of the compounds represented by the following structural formulas:

[0019]

[0020]

[0021] In a second aspect, the present invention provides a method for preparing the compound containing benzothiazole or benzoxazole described in the aforementioned embodiment, and the reaction is carried out according to the following synthetic route:

[0022]

[0023] Preferably, the compound represented by Formula 1, the compound represented by Formula 2 and a solvent are mixed and reacted at 90-120° C.;

[0024] Preferably, the molar ratio of the compound represented by Formula 1 to the compound represented by Formula 2 is 1:1.2-1.8.

[0025] In a third aspect, the present invention provides a use of the benzothiazole- or benzoxazole-containing compound described in the aforementioned embodiment in the preparation of a soluble epoxide hydrolase inhibitor.

[0026] The present invention has the following beneficial effects: The embodiments of the present invention provide new compounds that can inhibit the activity of soluble epoxide hydrolase and can be used as soluble epoxide hydrolase inhibitors to have a certain therapeutic effect on diseases caused by soluble epoxide hydrolase disorders. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] 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.

[0028] Figure 1 The results of the initial screening provided by the test example of the present invention;

[0029] Figure 2-Figure 3 The result diagram of the re-screening provided for the test example of the present invention;

[0030] Figure 4 The NMR carbon spectrum of the compound containing benzoxazole provided in Example 2 of the present invention;

[0031] Figure 5 This is the H NMR spectrum of the compound containing benzoxazole provided in Example 2 of the present invention;

[0032] Figure 6 This is the mass spectrum of the compound containing benzoxazole provided in Example 2 of the present invention. DETAILED DESCRIPTION

[0033] 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.

[0034] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0035] In a first aspect, the present invention provides a compound containing benzothiazole or benzoxazole, the structural formula of which is shown below:

[0036] Wherein, Y represents O or S, X represents hydrogen or halogen, and R represents any one of substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted C1-C10 alkyl and substituted or unsubstituted C3-C10 alkenyl.

[0037] For example, when Y is O, X represents halogen, and R represents any one of C2-C6 unsubstituted alkyl, C4-C6 unsubstituted cycloalkyl, substituted and unsubstituted phenyl; for example, halogen represents chlorine or bromine; the substituted phenyl is a monosubstituted phenyl; more preferably, it is a para-substituent.

[0038] Specifically, when Y is O and X represents chlorine, R represents a C2-C6 unsubstituted straight-chain alkyl group, a phenyl group, a C6-C7 unsubstituted cycloalkyl group, or a monosubstituted phenyl group, wherein the substituent of the monosubstituted phenyl group is selected from a C1-C5 carbonyl group; for example, R represents an unsubstituted straight-chain alkyl group such as ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, or n-heptyl, preferably a C3-C6 unsubstituted straight-chain alkyl group. The substituted phenyl group is a para-monosubstituted phenyl group, wherein the substituent is selected from a C1-C5 carbonyl group, preferably a C1-C3 carbonyl group. The cycloalkyl group is an unsubstituted cycloalkyl group such as cyclohexane or cycloheptane.

[0039] When Y is O and X represents bromine, R represents a C2-C6 unsubstituted straight-chain alkyl group, a C5-C7 unsubstituted cycloalkyl group, or a monosubstituted phenyl group, and the substituent of the monosubstituted phenyl group is selected from a C1-C5 ether group; for example, R represents an unsubstituted straight-chain alkyl group such as ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, or n-heptyl, preferably a C3-C6 unsubstituted straight-chain alkyl group, and the monosubstituted phenyl group is preferably an unpara-monosubstituted phenyl group, and the substituent of the para-monosubstituted phenyl group is selected from a C1-C5 ether group, preferably a C1-C3 ether group.

[0040] When Y is S, X represents bromine and chlorine, and R represents C1-C10 unsubstituted alkyl and monosubstituted phenyl; for example, R represents C1-C8 unsubstituted linear alkyl and para-monosubstituted phenyl, and the substituent is selected from C1-C5 unsubstituted linear alkyl.

[0041] Specifically, when Y is S and X represents bromine, R represents a monosubstituted phenyl group; the substituent of the monosubstituted phenyl group is selected from C1-C10 unsubstituted alkyl groups; preferably C1-C8 unsubstituted linear alkyl groups; more preferably C5-C8 unsubstituted linear alkyl groups.

[0042] When Y is S and X represents chlorine, R represents a para-monosubstituted phenyl group, and the substituent of the para-monosubstituted phenyl group is selected from C1-C5 unsubstituted straight-chain alkyl groups; preferably C1-C3 unsubstituted straight-chain alkyl groups.

[0043] More specifically, the compound containing benzothiazole or benzoxazole is selected from any one of the compounds represented by the following structural formulas:

[0044]

[0045]

[0046] In a second aspect, the present invention provides a method for preparing the compound containing benzothiazole or benzoxazole described in the aforementioned embodiment, and the reaction is carried out according to the following synthetic route:

[0047] The compound represented by Formula 1, the compound represented by Formula 2 and a solvent are mixed and reacted at 90-120° C.; the molar ratio of the compound represented by Formula 1 to the compound represented by Formula 2 is 1:1.2-1.8.

[0048] In a third aspect, the present invention provides the use of the benzothiazole- or benzoxazole-containing compound described in the aforementioned embodiments for the preparation of a soluble epoxide hydrolase inhibitor. Because the compound can function as a soluble epoxide hydrolase inhibitor, it has a therapeutic effect on diseases caused by soluble epoxide hydrolase imbalance, such as glioma, hypertension, renal deterioration, vascular inflammation, obstructive pulmonary disease, interstitial lung disease, and asthma.

[0049] Example 1

[0050] An embodiment of the present invention provides a benzoxazole-containing compound, the structural formula of which is shown below:

[0051] The preparation method is as follows:

[0052] Add 100 mg of 2-amino-5-bromobenzoxazole (about 0.47 mmol, 1 equivalent) to a 25 ml reaction bottle, then add 1.5 equivalents of cyclohexyl isocyanate, add about 10 ml of toluene to dissolve the reactants, place it on a heated magnetic stirrer, 120 degrees Celsius, 2000 rpm, and react for 12 hours.

[0053] After the reaction is completed, water is added to quench the reaction, and the mixture is extracted with ethyl acetate several times, an appropriate amount of saturated aqueous common salt is added, and Ph is adjusted to neutral. The organic layer is collected and distilled under reduced pressure to obtain a crude product. The crude product is pulped and purified with methanol. The crude product is placed in a 25ml reaction flask, an appropriate amount of methanol is added to dissolve, and the mixture is stirred at room temperature for 24h on a magnetic stirrer. TCL monitoring is performed. After stirring is completed, the liquid in the reaction flask is filtered by suction, and the precipitate is collected to obtain the product 1-(5-bromobenzoxazole-2-urea)-3-cyclohexyl.

[0054] The characterization data of the compound are as follows: 1 HNMR(600MHz,DMSO-d)6 11.12(s,1H),7.68(d,J=2.1Hz,1H),7.49(d,J=8.6Hz.1H),7.33(dd J=8 5, 2.2Hz, 1H), 1.83 (dtdd, J = 11.0, 5.5, 2.7, 1.4Hz, 2H), 1.71-1.61 (m2H), 1.55-1.48 (m, 1H), 1.38-1.10 (m, 5H). HRMS(ESI):calcdfor C 14 H 16 BrN3O2:337.0426; found:338.0504. 13 C NMR (101MHz, DMSO-d6) δ158.64,151.30,146.68,142.72,125.90,120.35,116.85,112.01,48.69,32.90,25.59,24.61.

[0055] Example 2

[0056] This embodiment provides a compound containing benzoxazole, the structural formula of which is shown below:

[0057]

[0058] This embodiment also provides a method for preparing the above-mentioned compound containing benzoxazole:

[0059] Add 100 mg of 2-amino-5-chlorobenzoxazole (about 0.6 mmol, 1 equivalent) to a 25 ml reaction bottle, then add 1.5 equivalents of trans-4-methylcyclohexyl isocyanate, add about 10 ml of toluene to dissolve the reactants, place it on a heated magnetic stirrer, 120 degrees Celsius, 2000 rpm, and react for 12 hours.

[0060] After reaction is completed, water is added to quench the reaction, and the mixture is extracted with ethyl acetate several times, and an appropriate amount of saturated aqueous common salt is added, and Ph is adjusted to neutral. The collected organic layer is distilled under reduced pressure to obtain a crude product. The crude product is beaten to purify the product with ethyl acetate. The crude product is placed in a 25ml reaction flask, an appropriate amount of ethyl acetate is added to dissolve, and the mixture is stirred at room temperature for 24h on a magnetic stirrer. TCL monitoring is performed. After stirring is completed, the liquid in the reaction flask is filtered by suction, and a precipitate is collected to obtain the product 1-(5-chlorobenzoxazole-2-urea)-3-trans-toluene.

[0061] The characterization spectrum of the benzoxazole-containing compound can be found in Figure 4-Figure 6 , the specific data are as follows: 1 HNMR(400MHz,DMSO-d)68.12(d,J=7.6Hz,1H),7.66-7.52(m,2H),7.24(dd J=8.6,2.2Hz,1H),3.38(s,57H),2.03-1.85(m,2H),1.70(d,J=12.9Hz,2H),1.42- 1.13(m4H),1.02(qd,J=13.3,3.3Hz,2H),0.89(d,J=6.5Hz.3H).HRMS(ESI):calcd for C 15 H 18 ClN3O2:307.1088; found:302.0683. 13 C NMR (101MHz, DMSO-d6) δ158.77,151.37,146.28,142.30,129.14,123.15,117.53,111.49,49.31,33.89,33.05,31.77,22.55.

[0062] Example 3-Example 60

[0063] Examples 3-60 The corresponding benzothiazole- or benzoxazole-containing compounds were synthesized according to the above method. Details will not be given here. Only the characterization results of the synthesized benzothiazole- or benzoxazole-containing compounds are provided, as follows:

[0064] Example 3—HO-1

[0065] Characterization data: 1H NMR(600MHz,DMSO-d6)δ10.98(s, 1 H),8.36(d,J=7.1Hz,1H),7.51(d,J=8.0Hz,1H),7.47(d,J=7.9Hz,1H),7.23(t,J=7.7Hz,1H),7.17(t,J=7.8Hz,1H),4.02(q,J=6 .8Hz,1H),1.90(dq,J=13.3,6.8Hz,2H),1.65(d,J=8.4Hz,2H),1.59-1.50(m,2H),1.44(dq,J=13.1,6.5Hz,2H).HRMS(ESI):calcd for C 13 H 15 N3O2:245.1164; found:246.1242. 13 C NMR (101MHz, DMSO-d6) δ155.43,153.28,147.63,142.58,125.13,123.98,114.94,109.65,56.46,32.56,23,78.

[0066] Example 4—HO-2

[0067] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.33(s,1H),10.36(s,1H),7.58(s,2H),7.48(d,J=8.2Hz,2H) ,7.27(d,J=28.8Hz,2H),6.93(d,J=8.3Hz,2H),3.74(s,3H).HRMS(ESI):(M+Na)+calcd for C 15 H 13 N3O3:306.0957; found:306.0865. 13 C NMR (101MHz, DMSO-d6) δ161.79,153.03,151.97,147.65,144.96,131.36,124.69,122.94,119.58,114.26,113.67,110.87,56.48.

[0068] Example 5—HO-3

[0069] Characterization data: 1H NMR (400MHz, DMSO-d6) δ11.50(s,1H),10.49(s,1H),7.96(d,J=8.1Hz,1H),7.56(dd,J=22.1,7.8Hz,2H),7 .30(t,J=7.5Hz,1H),7.20(tq,J=13.5,7.1,6.1Hz,3H),7.08-6.98(m,1H),2.37(s,4H).HRMS(ESI):calcd for C 15 H 13 N3O2:267.1008; found:268.1077. 13 C NMR (101MHz, DMSO-d6) δ153.45,152.24,149.78,143.91,135.67,132.02,131.21,126.56,125.45,124.32,115.78,114.25,109.87,17.23.

[0070] Example 6—HO-4

[0071] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.08(s,1H),8.43(s,1H),7.54(dd,J=17.2,7.8Hz,2H),7.25(dt, J=27.8,7.6Hz,2H),3.70(t,J=6.5Hz,2H),1.99(p,J=6.6Hz,2H).HRMS(ESI):(M+Na)+calcd for C 11 H 12 ClN3O2:276.0168; found:276.0510. 13 C NMR (101MHz, DMSO-d6) δ156.26,152.69,148.43,144.87,125.43,123.86,116.73,110.28,42.46,39.04,32.57.

[0072] Example 7—HO-5

[0073] Characterization data: 1H NMR(600MHz,DMSO-d6)δ11.31(s,1H),10.33(s,1H),7.82-7.36(m,4H),7.28(dt,J=15 .4,8.2Hz,1H),7.20(d,J=7.8Hz,1H),6.89(d,J=8.4Hz,2H).HRMS(ESI):(M+Na)+calcd for C 15 H 10 F3N3O3:360.0674; found:360.0574. 13 C NMR (101MHz, DMSO-d6) δ163.53,153.76,151.94,148.57,143.18,137.57,130.42,129.86,125.41,124.71,115.68,114.54,110.87,110.13.

[0074] Example 8—HO-6

[0075] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.19(s,1H),8.81(t,J=5.8Hz,1H),7.57(d,J=7.8Hz,1H),7.50(d,J=7.5 Hz,1H),7.36(d,J=4.4Hz,4H),7.31-7.17(m,3H),4.50(d,J=6.0Hz,2H).HRMS(ESI):(M+Na)+calcd for C 15 H 13 N3O2:290.1008; found:290.0896. 13 C NMR (101MHz, DMSO-d6) δ155.87,152.43,148.78,142.54,139.85,130.32,127.43,126,98,125.34,124.53,115.79,110.76,43.14.

[0076] Example 9—HO-7

[0077] Characterization data: 1H NMR (400MHz, DMSO-d6) δ10.70(s,1H),9.05(s,1H),7.92(d,J=21.1Hz,1H),7.66(s,1H),7. 40(s,1H),7.34-7.29(m,2H),7.14(d,J=8.1Hz,2H),2.26(s,3H).HRMS(ESI):(M+Na)+calcd for C 15 H 13 N3O2:290.1008; found:290.0905. 13 C NMR (101MHz, DMSO-d6) δ154.31,152,56,148.74,137.45,136.96,130.25,125.32,124.12,121.58,115.71,110.41,22.56.

[0078] Example 10—HO-8

[0079] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.15(s,1H),8.52(d,J=5.5Hz,1H),7.55(dd,J=15.7,7.8Hz,2H),7.26(dtd,J=26.4,7.6,1.2Hz,2H),5.9 4(ddd,J=22.1,10.3,5.1Hz,1H),5.29-5.19(m,1H),5.14(dd,J=10.3,1.7Hz,1H),3.93(d,J=5.7Hz,2H).HRMS(ESI):(M+Na)+calcd for C 11 H 11 N3O2:240.0851; found:240.0744. 13 C NMR (101MHz, DMSO-d6) δ154.87,152.92,149.65,144.57,134.28,125.79,124.93,117.41,114.32,111.56,44.84.

[0080] Example 11—HO-9

[0081] Characterization data: 1H NMR(600MHz, DMSO-d6)δ11.20(s,1H),8.81(s,1H),7.55(d,J=8.0Hz,1H),7.49(d,J=7.8Hz,1H),7.35(d,J=4.4Hz,4H),7.29-7.24(m,2H).HRMS(ESI):calcd for C 15 H 10 F3N3O2:321.0725; found:322.0798. 13 C NMR (101MHz, DMSO-d6) δ154.53,153.69,149.57,144.43,142.94,132.58,125.91,124.86,123.97,122.46,116.67,111.31.

[0082] Example 12—CO-2

[0083] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ11.15(s,1H),8.25(d,J=7.0Hz,1H),7.62-7.55(m,2H),7.23(dd,J=8.5,2.3Hz,1H),4.05(h,J=6.7Hz,1H), 1.93(dq,J=12.5,6.7Hz,2H),1.72-1.66(m,2H),1.58(tt,J=6.4,3.6Hz,2H),1.47(ddd,J=13.2,6.7,2.1Hz,2H).HRMS(ESI):calcd for C 13 H 14 ClN3O2:279.0775; found:280.0852. 13 C NMR (101MHz, DMSO-d6) δ154.81,152.25,146.94,144.46,129.82,123.57,118.76,111.45,59.45,32.84,23.68.

[0084] Example 13—CO-3

[0085] Characterization data: 1H NMR (400MHz, DMSO-d6) δ11.67(s,1H),10.54(s,1H),7.82(d,J=8.4Hz,2H),7.70( d,J=8.5Hz,2H),7.66-7.45(m,2H),7.27(dd,J=8.6,2.2Hz,1H).HRMS(ESI):calcd for C 15 H9Cl3N3O2:355.0335; found:356.0417. 13 C NMR (101MHz, DMSO-d6) δ153.58,152.46,146.84,145.73,142.86,133.58,129.43,126.31,124.51,122.05,120.85,117.67,110.43.

[0086] Example 14—CO-4

[0087] Characterization data: 1 H NMR(400MHz, DMSO-d6)δ11.33(s,1H),8.66(s,1H),7.65-7.50(m,2H),7.40-7.20(m,6H),4.49(d,J=6.0Hz,2H).HRMS(ESI):(M+Na)+calcd forC 15 H 12 ClN3O2:324.0618; found:324.0509. 13 C NMR (101MHz, DMSO-d6) δ154.34,152.45,147.85,145.93,138.43,128.75,127.56,123.41,119.58,109.91,45.31.

[0088] Example 15—CO-5

[0089] Characterization data: 1 H NMR(400MHz, DMSO-d6)δ10.96(s,1H),8.21(s,1H),7.57(dd,J=5.4,3.2Hz,2H),7.23(dd,J=8.6,2.2Hz,1H),1.37(s,9H).HRMS(ESI):(M+Na)+calcd forC 12 H 14 ClN3O2:290.0775; found:290.0664. 13C NMR (101MHz, DMSO-d6) δ155.43,153.45,147.69,145.94,129.23,122.47,118.64,110.47,57.31,29.13.

[0090] Example 16—CO-6

[0091] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ11.29(s,1H),8.35(dt,J=10.2,5.7Hz,1H),7.62-7.58(m,2H),7.25(dd,J=8.6,2.1Hz,1H),5.91(ddt,J=17.3 ,10.3,5.1Hz,1H),5.22(dq,J=17.2,1.7Hz,1H),5.13(dq,J=10.5,1.5Hz,1H),3.90(tt,J=5.5,1.8Hz,2H).HRMS(ESI):(M+Na)+calcd for C 11 H 10 ClN3O2:274.0462; found:274.0350. 13 C NMR (101MHz, DMSO-d6) δ155.32,153.22,147.24,145.31,135.64,128.53,122.47,117.54,109.89,44.53.

[0092] Example 17—CO-7

[0093] Characterization data: 1 H NMR(600MHz,DMSO-d6)δ8.18(dt,J=11.7,5.8Hz,1H),7.50-7.46(m,2H),7.14(dd,J=8.4,2.4 Hz,1H),3.60(t,J=6.3Hz,2H),3.26(d,J=6.4Hz,2H),1.88(p,J=6.5Hz,2H).HRMS(ESI):calcd for C 11 H 11 Cl2N3O2:287.0228; found:288.0306. 13 C NMR (101MHz, DMSO-d6) δ154.43,152.65,147.78,145.53,128.43,122.57,118.91,110.21,42.46,30.14,32.46.

[0094] Example 18—CO-8

[0095] Characterization data: 1 H NMR(400MHz, DMSO-d6)δ11.16(s,1H),8.23(t,J=5.5Hz,1H),7.63-7.53(m,2H),7.23(dd,J=8.5,2.2Hz,1H), 3.23(q,J=6.7Hz,2H),1.49(q,J=7.1Hz,2H),1.41-1.14(m,6H),0.92-0.80(m,3H).HRMS(ESI):(M+Na)+calcd for C 14 H 18 ClN3O2:318.1088; found:318.0988. 13 C NMR (101MHz, DMSO-d6) δ158.82,152.20,146.27,142.32,129.12,123.09,117.34,111.47,31.38,29.75,26.45,22.50,14.33.

[0096] Example 19—CO-9

[0097] Characterization data: 1 H NMR(400MHz, DMSO-d6)δ11.66(s,1H),10.35(s,1H),7.65(d,J=9.5Hz,2H),7.40-6.90(m,5H),2.39(s,3H).HRMS(ESI):(M+Na)+calcd for C 15 H 12 ClN3O2:324.0618; found:324.0508. 13 C NMR(101MHz,DMSO-d6)δ153.43,152.67,146.78,145.69,135.67,132.46, 130.32,129.87,128.79,126.43,123.54,117.68,115.92,110.21,18.02.

[0098] Example 20—CO-10

[0099] Characterization data: 1H NMR(600MHz, DMSO-d6)δ11.64(s,1H),10.38(s,1H),7.65(dd,J=56.9,6.7Hz,4H),7.35(d,J=8.5Hz,2H),7.27(dd,J=8.6,2.2Hz,1H).HRMS(ESI):(M+Na)+calcd for C 15 H9ClF3N3O3:394.0285; found:394.0176. 13 C NMR (101MHz, DMSO-d6) δ161.21,153.32,152.58,147.53,145.97,137.56,130.47,128.34,123.86,117.48,114.23,111.04,109.84.

[0100] Example 21—CO-11

[0101] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.66(d,J=148.5Hz,1H),8.30(d,J=8.3Hz,1H),7.97(d,J=1.6Hz,1H),7.94(d,J=1.7Hz,1H),7.82-7.75(m, 1H),7.67(d,J=2.9Hz,1H),7.58(s,1H),7.23(d,J=2.2Hz,1H),7.02(d,J=1.3Hz,1H),7.00(dd,J=3.0,1.8Hz,1H).HRMS(ESI):calcd for C 14 H9ClN4O4:332.0312; found:333.0377. 13 C NMR (101MHz, DMSO-d6) δ153.41,152.79,146.54,145.37,142.68,131.21,128.92,125.57,123.46,118,56,117.23,111.14.

[0102] Example 22—CO-12

[0103] Characterization data: 1H NMR (400MHz, DMSO-d6) δ11.14(s,1H),8.23(d,J=7.7Hz,1H),7.64-7.51(m,2H),7.24(dd,J=8.6,2.2Hz,1H),3.62(s,1H),1. 87(d,J=11.4Hz,2H),1.75-1.64(m,2H),1.55(d,J=13.0Hz,1H),1.30(tt,J=20.9,10.9Hz,5H).HRMS(ESI):(M+Na)+calcdfor C 14 H 16 ClN3O2:316.0931; found:316.0815. 13 C NMR (101MHz, DMSO-d6) δ154.57,152.46,147.31,145.54,129.34,122.43,117.68,111.07,53.89,32.59,26.25,24.58.

[0104] Example 23—CO-13

[0105] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ10.22 (s, 1H), 7.56 (td, J = 5.0, 2.4Hz, 4H), 7.22 (dd, J = 8. 6,2.2Hz,1H),7.16(q,J=8.8,8.0Hz,2H),3.31(s,3H).HRMS(ESI):(M+Na)+calcd for C 16 H 12 ClN3O3:352.0567; found:352.0466. 13 C NMR (101MHz, DMSO-d6) δ198.13,152.58,151.67,147.32,145.87,143.89,137.47,129.54,128.86,123.43,121.58,117.68,110.24,28.03.

[0106] Example 24—BO-1

[0107] Characterization data: 1H NMR(400MHz, DMSO-d6)δ11.00(s,1H),7.86(d,J=8.4Hz,1H),7.82(s,1H),7.40-7.32(m,5H),7.27(s,1H),4.38(d,J=5.9Hz,2H).HRMS(ESI):(M+Na)+calcdfor C 15 H 12 BrN3O2:368.0113; found:368.0000. 13 C NMR (101MHz, DMSO-d6) δ154.67,152.78,147.41,144.97,138.23,129.43,127.51,126.82,124.53,120.76,123.57,44.67.

[0108] Example 25—BO-2

[0109] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ11.16(s,1H),8.12(d,J=7.6Hz,1H),7.72(d,J=2.0Hz,1H),7.53(d,J=8.6Hz,1H),7.36(dd,J=8.6,2.0Hz,1H),3.56-3.46(m,1 H),1.97-1.89(m,2H),1.76-1.64(m,2H),1.43-1.32(m,1H),1.26(qd,J=12 .7,3.4Hz,2H),1.07-0.95(m,2H),0.88(d,J=6.5Hz,3H).HRMS(ESI):calcd for C 15 H 18 BrN3O2:351.0582; found:368.0003. 13 C NMR (101MHz, DMSO-d6) δ154.67,152.46,147.78,143.61,128.34,124.53,120.14,113.46,53.68,33.57,32.12,31.23,21.04.

[0110] Example 26—BO-4

[0111] Characterization data: 1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),9.35(s,1H),7.96(s,1H),7.73(s,1H),7 .64(s,1H),7.39-7.30(m,3H),7.28(d,J=1.6Hz,1H).HRMS(ESI):(M+Na)+calcd for C 15 H9BrF3N3O3:437.9779; found:437.9666. 13 C NMR (101MHz, DMSO-d6) δ161.13,152.43,151.35,147.49,144.75,137.53,130.23,128.93,127.03,124.25,120.41,114.58,112.98,110.58.

[0112] Example 27—BO-5

[0113] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.47(s,1H),10.31(s,1H),7.58(s,1H),7.47(d,J=7.9Hz,2H),7.40(d,J =8.5Hz,1H),7.32(d,J=8.3Hz,1H),7.17(d,J=8.1Hz,2H),1.99(s,3H).HRMS(ESI):(M+Na)+calcd for C 15 H 12 BrN3O2:368.0113; found:368.0011. 13 C NMR (101MHz, DMSO-d6) δ153.31,152.12,147.45,143.96,137.18,136.57,129.89,128.01,124.67,121.43,120.29,113.06,21.93.

[0114] Example 28—BO-6

[0115] Characterization data: 1H NMR (400MHz, DMSO-d6) δ11.45(s,1H),10.20(s,1H),7.76(s,1H),7.55(d,J=8.3Hz,1H),7.46(d ,J=8.5Hz,2H),7.41-7.29(m,1H),6.92(d,J=8.5Hz,2H),3.74(s,3H).HRMS(ESI):(M+Na)+calcd for C 15 H 12 BrN3O3:383.0062; found:383.0057. 13 CNMR(101MHz,DMSO-d6)δ159.45,152.97,152.13,148.34,143.31,132.87,128.76,124.58,120.21,114.37,113.32,56.24.

[0116] Example 29—BO-7

[0117] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ11.22(s,1H),8.28(s,1H),7.69(s,1H),7.53(d,J=8.5Hz,1H),7.36(dd,J=8.6,2 .0Hz,1H),3.69(t,J=6.5Hz,2H),3.37(d,J=6.5Hz,2H),1.97(p,J=6.7Hz,2H).HRMS(ESI):(M+Na)+calcd for C 11 H 11 BrClN3O2:353.9723; found:353.0917. 13 C NMR (101MHz, DMSO-d6) δ155.32,153.59,147.96,143.68,128.76,124.41,120.68,113.48,41.68,40.34,32.25.

[0118] Example 30—BO-8

[0119] Characterization data: 1H NMR (600MHz, DMSO-d6) δ11.29(s,1H),8.36(t,J=5.9Hz,1H),7.75-7.72(m,1H),7.55(d,J=8.6Hz,1H),7.37(dd,J=8.6,2.0Hz,1H),5.91(ddt ,J=17.2,10.3,5.0Hz,1H),5.22(dq,J=17.2,1.7Hz,1H),5.13(dq,J=10.3,1.6Hz,1H),3.90(tt,J=5.5,1.7Hz,2H).HRMS(ESI):(M+Na)+calcd for C 11 H 10 BrN3O2:317.9956; found:317.9839. 13 C NMR (101MHz, DMSO-d6) δ154.24,153.15,148.17,143.38,134.31,128.43,124.04,120.13,117.48,113.87,44.68.

[0120] Example 31—BO-9

[0121] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ10.29(s,1H),7.88(dd,J=8.7,1.7Hz,3H),7.71(d,J=2.0Hz,1H),7.7 0(d,J=1.8Hz,1H),7.64-7.62(m,1H),7.57(dd,J=9.2,2.3Hz,1H).HRMS(ESI):(M+Na)+calcd for C 15 H 12 BrN3O2:368.0113; found:368.0005. 13 C NMR(101MHz,DMSO-d6)δ152.97,152.08,147.79,143.56,135.68,131.79, 130.69,129.68,128.42,126.89,124.31,120.56,115.49,113.25,17.14.

[0122] Example 32—BO-10

[0123] Characterization data: 1H NMR (600MHz, DMSO-d6) δ11.18-11.13(m,1H),8.25(d,J=7.0Hz,1H),7.72(hept,J=1.4Hz,1H),7.53(dp,J=8.6,1.5Hz,1H),7.38-7. 34(m,1H),4.08-4.00(m,1H),1.96-1.88(m,2H),1.73-1.63(m,2H),1.63-1.53(m,2H),1.51-1.42(m,2H).HRMS(ESI):(M+Na)+calcd for C 13 H 14 BrN3O2:346.0269; found:346.0613. 13 C NMR (101MHz, DMSO-d6) δ154.97,152.76,147.64,143.13,128.39,124.61,119.86,112.47,58.34,32.42,23.31.

[0124] Example 33—BO-11

[0125] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ11.20(s,1H),8.23(t,J=5.8Hz,1H),7.67(s,1H),7.51(d,J=8.4Hz,1H),7.36-7.32(m,1H) ,3.22(q,J=6.8Hz,2H),1.50(p,J=7.1Hz,2H),1.32-1.24(m,7H),0.87(d,J=6.9Hz,3H).HRMS(ESI):(M+Na)+calcd forC 14 H 18 BrN3O2:362.0582; found:362.0476. 13 C NMR (101MHz, DMSO-d6) δ155.46,153.12,148.01,143.68,127.94,124.03,120.45,113.76,20.68,31.34,30.15,26.75,23.52,14.68.

[0126] Example 34—BO-12

[0127] Characterization data: 1H NMR (400MHz, DMSO-d6) δ11.33(s,1H),9.53(d,J=129.0Hz,1H),8.11(d,J=19.1Hz,1H) ,7.88(s,2H),7.78(s,1H),7.68(s,1H),7.48-7.37(m,1H).HRMS(ESI):(M+Na)+calcd for C 15 H8BrClF3N3O2:455.9441; found:455.9338. 13 C NMR (101MHz, DMSO-d6) δ152.31,151.78,148.59,143.73,134.58,129.53,128.67,127.84,134.31,133.58,119.79,119.03,113.57.

[0128] Example 35—BO-13

[0129] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ10.33(s,1H),7.91(s,1H),7.74-7.68(m,2H),7.61(d,J=4.5Hz,1H), 7.57(d,J=8.8Hz,1H),7.43(d,J=8.5Hz,1H),7.28(d,J=8.2Hz,1H).HRMS(ESI):(M+Na)+calcd for C 16 H 12 BrN3O3:395.0062; found:395.9960. 13 CNMR(101MHz,DMSO-d6)δ197.45,152.89,151,68,148.51,144.03,137.75,129.43,127.59,124.79,122.01,120.85,113.42,27.13.

[0130] Example 36—HS-1

[0131] Characterization data: 1H NMR (400MHz, DMSO-d6) δ10.80(s,1H),9.21(s,1H),8.66(s,1H),7.89(d,J=7.9Hz,1H),7.64(s,1 H),7.53(dd,J=8.9,5.1Hz,2H),7.47-7.37(m,2H),7.17-7.11(m,2H).HRMS(ESI):(M+Na)+calcd for C 14 H 10 FN3OS:310.0529; found:310.0415. 13 CNMR(101MHz,DMSO-d6)δ174.79,163.47,153.57,152.49,135.72,131.47,125.68,124.52,122.58,119.68,118.47,115.83.

[0132] Example 37—HS-2

[0133] Characterization data: 1 H NMR(400MHz, DMSO-d6)δ9.58(s,1H),9.22(s,2H),7.95(s,1H),7.67(d,J=8.7Hz,3H),7.60(s,2H),7.59(d,J=1.9Hz,2H).HRMS(ESI):(M+Na)+calcd forC 16 H 13 N3O2S:334.0728; found:334.0611. 13 C NMR (101MHz, DMSO-d6) δ198.23,174.34,153.79,152.45,148.21,136.79,131.45,129.13,125.89,124.57,121.79,121.13,118.67,27.46.

[0134] Example 38—HS-3

[0135] Characterization data: 1H NMR (600MHz, DMSO-d6) δ9.55(s,1H),9.19(s,1H),7.92(d,J=1.9Hz,1H),7.91(d,J=2.0Hz,1H),7.90-7.86(m,2H),7.65(d, J=8.4Hz,2H),7.37(ddd,J=8.3,7.2,1.2Hz,1H),7.22(ddd,J=8.1,7.3,1.1Hz,1H),2.50(s,3H).HRMS(ESI):(M+Na)+calcd for C 15 H 13 N3OS:306.0779; found:306.0675. 13 C NMR (101MHz, DMSO-d6) δ175.21,153.79,152.57,137.03,136.47,131.59,129.31,125.46,124.39,121.47,119.34,21.78.

[0136] Example 39—HS-4

[0137] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.11(s,1H),8.31(d,J=8.4Hz,1H),8.13(dd,J=8.4,1.6Hz,1H),8.03(dd,J=8.3,1 .6Hz,1H),7.80-7.73(m,1H),7.70(ddd,J=8.6,7.3,1.6Hz,1H),7.28(tt,J=8.1,7.0Hz,3H).HRMS(ESI):(M+Na)+calcdfor C 14 H 10 N4O3S:337.0474; found:337.0362. 13 C NMR (101MHz, DMSO-d6) δ174.43,153.87,151.96,142.57,131.78,125.46,124.76,122.35,118.75.

[0138] Example 40—HS-5

[0139] Characterization data: 1H NMR (400MHz, DMSO-d6) δ11.12(s,1H),8.67(s,1H),7.93(d,J=7.9Hz,1H),7.87(d,J=8.1Hz,1H),7.69 (d,J=8.1Hz,1H),7.45-7.37(m,1H),7.29-7.20(m,3H),7.08-7.01(m,1H).HRMS(ESI):(M+Na)+calcd for C 15 H 13 N3OS:306.0779; found:306.0673. 13 C NMR (101MHz, DMSO-d6) δ175.43,153.78,152.47,135.46,132.48,130.85,129.42,125.78,125.14,124.68,122.51,118.48,115.47,17.31.

[0140] Example 41—HS-6

[0141] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ10.72(s,1H),9.03(s,1H),8.44(s,1H),7.92-7.79(m,1H),7.63(dd,J=20.3 ,8.1Hz,1H),7.30-7.26(m,2H),7.10(d,J=8.2Hz,2H),7.06-6.98(m,2H).HRMS(ESI):(M+Na)+calcd for C 15 H 10 F3N3OS:360.0497; found:360.0393. 13 C NMR (101MHz, DMSO-d6) δ175.23,153.58,151.93,143.57,132.46,131.86,125.68,124.69,122.35,118.69.

[0142] Example 42—HS-7

[0143] Characterization data: 1H NMR(400MHz,DMSO-d6)δ10.75(s,1H),9.17(s,1H),7.89(s,1H),7.65(s,1H),7.41 -7.32(m,6H),7.24(d,J=7.6Hz,1H),7.02-6.97(m,4H).HRMS(ESI):(M+Na)+calcd for C 20 H 15 N3O2S:384.0885; found:384.0777. 13 C NMR(101MHz,DMSO-d6)δ174.98,157.47,153.46,152.36,150.48,132.47, 131.58,128.48,125.79,124.45,122.42,121.56,119.46,118.57,115.89.

[0144] Example 43—HS-8

[0145] Characterization data: 1 H NMR(400MHz,DMSO-d6)δ10.69(s,1H),8.97(s,1H),7.90(s,1H),7.65(s,1H),7.49 -7.28(m,4H),6.91(d,J=8.5Hz,2H),3.79-3.72(m,3H).HRMS(ESI):(M+Na)+calcd for C 15 H 13 N3O2S:322.0728; found:322.0625. 13 C NMR (101MHz, DMSO-d6) δ174.47,159.32,153.89,152.31,132.41,131.62,125.68,124.15,122.41,120.34,118.67,114.31,56.14.

[0146] Example 44—CS-1

[0147] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ10.97(s,1H),9.20(s,1H),7.96(d,J=8.5Hz,1H),7.74(s,1H),7.53( d,J=6.6Hz,2H),7.30(dd,J=8.4,2.1Hz,1H),7.19(t,J=8.9Hz,2H).HRMS(ESI):(M+Na)+calcd for C14 H9ClFN3OS:344.0319; found:344.0035. 13 CNMR(101MHz,DMSO-d6)δ175.21,163.34,152.14,150.46,135.46,130.83,129.54,124.58,123.42,121.89,119.49,116.26.

[0148] Example 45—CS-2

[0149] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ9.62(s,1H),8.10(s,1H),7.90(d,J=8.4Hz,1H),7.75(d,J =8.9Hz,1H),7.63(d,J=8.8Hz,2H),7.26(dd,J=8.4,2.1Hz,1H).HRMS(ESI):calcd for C 15 H8Cl2F3N3OS:404.9717; found:405.9784. 13 C NMR (101MHz, DMSO-d6) δ174.68,152.41,150.52,134.78,130.47,129.52,128.93,124.57,123.76,121.48,119.69.

[0150] Example 46—CS-3

[0151] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.17(s,1H),8.79(t,J=5.8Hz,1H),7.55(d,J=7.8Hz,1H),7.49(d,J=7.5Hz,1H),7.35(s,1H),7.3 4(s,2H),7.26(td,J=6.0,4.7,2.0Hz,2H),7.21(dd,J=7.6,1.2Hz,1H),4.49(d,J=6.0Hz,2H).HRMS(ESI):(M+Na)+calcdfor C 15 H 12 ClN3OS:340.0390; found:340.0290. 13C NMR (101MHz, DMSO-d6) δ175.13,154.57,150.78,138.13,130.59,128.98,128.42,126.87,126.31,124.25,123.67,121.56,44.79.

[0152] Example 47—CS-4

[0153] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ10.55(s,1H),7.90(d,J=8.4Hz,1H),7.65(s,1H),7.23(dd,J=8.4,2.0Hz,1H),6.61(s,1H),3.99(h,J=6.5Hz,1H),1.87(d,J= 12.4Hz,2H),1.68(d,J=12.8Hz,2H),1.33(s,1H),1.19(q,J=12.2,11.0Hz,2H),1.00(q,J=13.1,12.7Hz,2H),0.87(d,J=6.5Hz,3H).HRMS(ESI):calcd for C 15 H 18 ClN3OS:323.0859; found:340.0273. 13 C NMR (101MHz, DMSO-d6) δ174.58,154.78,150.41,130.69,129.21,124.57,123.45,121.15,54.68,33.42,32.53,31.79,20.52.

[0154] Example 48—CS-5

[0155] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),9.35(s,1H),7.96(s,1H),7.73(s,1H),7.64(s,1H),7.39-7.30(m,3H),7.28(d,J=1.6Hz,1H).HRMS(ESI):calcd forC 15 H9ClF3N3O2S:387.0056; found:388.0130. 13C NMR (101MHz, DMSO-d6) δ175.13,161.59,152.57,150.16,137.57,130.68,129.89,128.93,124.32,124.56,121.71,114.35,110.43,109.78.

[0156] Example 49—CS-6

[0157] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ10.52(s,1H),7.91(d,J=8.4Hz,1H),7.67(d,J=2.0Hz,1H),7.25(dd,J=8.4,2.1Hz,1H),6.77(d,J=7.2Hz,1H ),4.00(h,J=7.2,6.7Hz,1H),1.95-1.83(m,2H),1.73-1.62(m,2H),1.60-1.52(m,2H),1.49-1.37(m,2H).HRMS(ESI):(M+Na)+calcd for C 13 H 14 ClN3OS:318.0546; found:318.0439. 13 C NMR (101MHz, DMSO-d6) δ175.31,154.37,150.97,130.53,129.32,124.67,123.59,121.95,58.45,32.43,23.31.

[0158] Example 50—CS-7

[0159] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),9.56(s,1H),7.95(d,J=8.7Hz,1H),7.75(s,1H) ),7.72(s,2H),7.69(d,J=8.7Hz,3H),7.29(dd,J=8.6,2.0Hz,1H).HRMS(ESI):calcd for C 15 H9ClF3N3OS:371.0107; found:372.0179. 13 C NMR (101MHz, DMSO-d6) δ174.57,152.14,150.87,143,24,132.58,130.41,129.59,125.68,124.58,123.79,122.32,121.36.

[0160] Example 51—BS-1

[0161] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.27(s,1H),8.56(s,1H),7.93-7.85(m,2H),7.81(d,J=8.1Hz,1H),7.40(dd,J=8.4,1.9 Hz,1H),7.23(d,J=7.8Hz,1H),7.19(d,J=8.2Hz,1H),7.04(td,J=7.4,1.3Hz,1H),2.26(s,3H).HRMS(ESI):calcd forC 15 H 12 BrN3OS:360.9884; found:361.9947. 13 C NMR (101MHz, DMSO-d6) δ174.45,152.14,151.34,135.46,131.57,130.46,129.89,129.31,125.98,125.31,124.43,116.35,114.76,17.21.

[0162] Example 52—BS-2

[0163] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ10.78(s,1H),7.85(d,J=8.4Hz,1H),7.80(d,J=1.9Hz,1H),7.36(dd,J=8.4,2.0Hz,1H) ,6.73(s,1H),4.10(d,J=5.2Hz,2H),1.53-1.42(m,2H),1.35-1.25(m,6H),0.93-0.84(m,3H).HRMS(ESI):calcd for C 14 H 18 BrN3OS:355.0354; found:356.0423. 13 C NMR (101MHz, DMSO-d6) δ174.98,154.31,151.57,129.98,129.21,125.67,124.54,116.73,40.43,31.35,30.68,26.13,23.15,14.35.

[0164] Example 53—BS-3

[0165] Characterization data:1 H NMR (400MHz, DMSO-d6) δ10.88(s,1H),7.86(d,J=8.4Hz,1H),7.80(d,J=1.9Hz,1H),7.36(dd,J=8.4, 1.9Hz,1H),6.86(s,1H),3.68(t,J=6.5Hz,2H),3.30(s,2H),1.94(p,J=6.6Hz,2H).HRMS(ESI):calcd for C 11 H 11 BrClN3OS:346.9495; found:347.9577. 13 C NMR (101MHz, DMSO-d6) δ175.14,154.53,151.74,129.89,128.98,125.43,124.14,116.57,41.57,39.87,32.13.

[0166] Example 54—BS-4

[0167] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.10(s,1H),9.55(s,1H),7.90(d,J=8.4Hz,2H),7.74(d,J=8.6Hz,2H),7.70(s,2H),7.41(dd,J=8.4,1.9Hz,1H).HRMS(ESI):calcd for C 15 H9BrF3N3OS:414.9602; found:415.9680. 13 C NMR (101MHz, DMSO-d6) δ175.14,152.32,151.68,143.57,132.46,129.92,129.13,125.43,124,35,122.73,116.58.

[0168] Example 55—BS-5

[0169] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ11.33(s,1H),9.69(s,1H),8.11(d,J=19.1Hz,1H),7.88(s,2H),7.78(s,2H),7.68(s,1H).HRMS(ESI):calcd forC 15 H8BrClF3N3OS:448.9212; found:449.9280. 13C NMR (101MHz, DMSO-d6) δ174.68,152.68,151.43,134.67,129.89,129.34,128.67,125.42,124.56,123.13,119.57,116.41.

[0170] Example 56—BS-6

[0171] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ10.90(s,1H),7.86(d,J=8.4Hz,1H),7.81(d,J=1.8Hz,1H),7.37(dd,J=8.4,1.9Hz,1H),6.87(s,1H) ,5.88(ddd,J=22.2,10.4,5.2Hz,1H),5.19(dd,J=17.2,1.8Hz,1H),5.15-5.08(m,1H),3.86-3.75(m,2H).HRMS(ESI):calcd for C 11 H 10 BrN3OS:310.9728; found:311.9809. 13 C NMR (101MHz, DMSO-d6) δ174.56,154.68,151.97,134.56,130.14,129.56,125.67,124.75,116.76,115.43,44.56.

[0172] Example 57—BS-7

[0173] Characterization data: 1 H NMR(400MHz, DMSO-d6)δ11.00(s,1H),7.86(d,J=8.4Hz,1H),7.82(s,1H),7.40-7.32(m,5H),7.27(s,1H),4.38(d,J=5.9Hz,2H).HRMS(ESI):calcd forC 15 H 12 BrN3OS:360.9884; found:361.9949. 13 C NMR (101MHz, DMSO-d6) δ175.31,154.53,151.67,138.68,130.14,129.57,128.78,126.42,125.73,124.35,116.75,44.68.

[0174] Example 58—BS-8

[0175] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ10.95(s,1H),9.16(s,1H),7.90(d,J=12.1Hz,2H),7.55(s,1H),7.53(s,1H),7.43-7.39 (m,2H),7.39-7.36(m,2H),7.13(d,J=7.5Hz,1H),7.03(d,J=8.9Hz,2H),6.99(d,J=7.5Hz,2H).HRMS(ESI):calcd forC 20 H 14 BrN3O2S:438.9990; found:440.0066. 13 C NMR (101MHz, DMSO-d6) δ174.79,157.51,152.34,151.57,149.88,132.44,129. 89,129.14,128.34,125.42,124.31,122.21,121.68,119.45,116.31,115.42.

[0176] Example 59—BS-9

[0177] Characterization data: 1 H NMR (400MHz, DMSO-d6) δ10.43(s,1H),7.84(d,J=8.4Hz,1H),7.79(d,J=1.9Hz,1H),7.35(dd,J=8.4,2.0Hz,1H),6.62(s,1H),1.33(s,9H).HRMS(ESI):calcd for C 14 H 12 BrN3OS:327.0041; found:328.0120. 13 C NMR (101MHz, DMSO-d6) δ174.67,154.44,151.79,130.11,129.56,125.54,124.32,116.57,57.32,29.03.

[0178] Example 60—BS-10

[0179] Characterization data: 1H NMR (600MHz, DMSO-d6) δ10.96 (s, 1H), 9.24-9.09 (m, 1H), 7.84 (d, J = 8.4Hz, 1H), 7.81 (s, 1H), 7.49 (dd,J=8.9,4.8Hz,2H),7.35(dd,J=8.4,1.9Hz,1H),7.19-7.09(m,2H).HRMS(ESI):(M+Na)+calcd for C 14 H9BrFN3OS:369.9634; found:365.9716. 13 C NMR (101MHz, DMSO-d6) δ174.54,163.89,152.13,151.21,135.67,130.14,129.54,125.54,124.67,119.56,116.73,115.46.

[0180] Test example

[0181] Epoxyeicosatrienoic acids (EETs) are products of arachidonic acid metabolism via cytochrome P450 enzymes and possess antihypertensive, fibrinolytic, and anti-inflammatory activities. EETs are rapidly hydrolyzed in vivo by soluble epoxide hydrolase (sEH), reducing their activity. sEH inhibitors can increase endogenous EET levels.

[0182] We established an enzymatic reaction system to screen test compounds for their activity against hsEH. The specific substrate, PHOME, is inherently nonfluorescent, but it undergoes hydrolysis under the action of hsEH to produce 6-methoxy-2-naphthaldehyde, which emits fluorescence at a wavelength of 465 nm when excited by 330 nm light. The approximate initial velocity of the enzymatic reaction is calculated by calculating the rate of change of fluorescence intensity per unit time. Comparison with blank control wells allows the determination of the compound's effect on the hsEH enzyme.

[0183] (1) Reagents used: physiological saline, DMSO, hsEH enzyme working solution (1 mg / ml)

[0184] Solution of the compound to be tested: According to the molecular mass and weight of the provided compound, a 10 mM stock solution was prepared with DMSO, and diluted with physiological saline to obtain the corresponding primary screening working solution (100 μM) and a series of rescreening working solutions.

[0185] Preparation of 25 mM HEPES buffer (containing 0.1 mg / ml BSA): Weigh 595.75 mg HEPES, dissolve it in 100 ml pure water, adjust the pH to 7.0 with 1 M NaOH, and fully dissolve 10 mg BSA therein.

[0186] PHOME substrate working solution (6.25uM): 5μL 5mM stock solution + 4000μL HEPES buffer, store at -20℃ in the dark.

[0187] AUDA stock solution (25 mM): 10.0 mg AUDA + 1019 μL DMSO will yield a 25 mM AUDA stock solution. Aliquot and store at -80°C. Dilute with saline or DMSO before use.

[0188] The solution settings for each well are as follows:

[0189]

[0190] (1) Initial screening

[0191] The initial screening reaction system was 50 μL, that is, the final enzyme concentration was 0.5 mg / mL, the substrate PHOME was 2.5 μM; the final concentration of the test compound was 10 μM; the final concentration of the 1 μM positive inhibitor AUDA reaction system was 100 nM, and the final concentration of DMSO was 1‰.

[0192] (2) Rescreening

[0193] The total volume, enzyme concentration, and substrate concentration of the reaction system remained unchanged. A 1 mM solution of the test compound was serially diluted to obtain a series of 300, 100, 30, 10, 3, and 1 μM working solutions for repeated screening, with final concentrations of 30, 10, 3, 1, 0.3, and 0.1 μM. The AUDA stock solution was diluted with DMSO and saline to obtain a series of 10, 3, 1, 0.3, 0.1, 0.03, and 0.01 μM AUDA inhibitor working solutions, with a maximum DMSO concentration of 3‰.

[0194] (3) Fluorescence intensity value detection

[0195] After adding enzyme working solution and the test compound, inhibitor, or saline to each well, incubate at 25°C for 15 minutes, then add substrate working solution. In kinetic monitoring mode, fluorescence intensity is measured every 2 minutes for 20 minutes. The difference in fluorescence intensity between 0 and 4 minutes is used to calculate the reaction rate for each well, representing the approximate initial velocity of the enzymatic reaction.

[0196] (4) Results and analysis

[0197] The initial reaction velocity of the sample test well is represented by V, the initial velocity of the background well is represented by Vb, and the initial velocity of the blank control well is represented by Vc. The inhibition rate of the compound on the enzyme is calculated.

[0198]

[0199] (4.1) Initial screening results

[0200] The results are shown in Table 1 and Figure 1 .

[0201] Table 1 Preliminary screening results

[0202]

[0203]

[0204] According to the initial screening results, 11 of the submitted compounds had an inhibition rate greater than 50%, indicating a certain inhibitory effect, and were therefore included in the subsequent rescreening.

[0205] (4.2) Rescreening results

[0206] After repeated screening, the IC values ​​of each compound on hsEH enzyme were determined. 50 The dose-effect curve was drawn and the results were shown in Table 2 and Figure 2-Figure 3 .

[0207] Table 2 Rescreening results

[0208]

[0209] It can be seen that the compounds provided in the embodiments of the present invention have excellent hsEH enzyme inhibition effects, which basically reach the effect of the positive control drug, and even the effects of most compounds are better than the positive control drug.

[0210] 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. Use of a compound containing benzothiazole or benzoxazole in the preparation of a soluble epoxide hydrolase inhibitor, characterized in that: The compound containing benzothiazole or benzoxazole is selected from any one of the compounds represented by the following structural formula: 、 、 、 、 、 、 、 、 、 and .

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