Benzothiazole coupled hydroxamic acid compound as well as preparation method and application thereof

Abstract

The invention relates to the field of medicines, in particular to a benzothiazole coupled hydroxamic acid compound containing a formula (I) and application of the benzothiazole coupled hydroxamic acid compound in preparation of a bacterial biofilm inhibitor. In-vivo and in-vitro pharmacological verification shows that most of the benzothiazole coupled hydroxamic acid compounds have excellent bacterial biofilm inhibition activity, can be used for preparing anti-biofilm agents, and have the potential of being developed into novel antibacterial drugs. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medicine, and particularly relates to a class of benzothiazole coupled hydroxamic acid compounds and a preparation method and application thereof. Background Art

[0002] The discovery of antibiotics has changed the trajectory of medical development, allowing humans to greatly reduce the harm caused by microbial infections. However, in recent decades, the overuse of antibiotics and social and economic factors have accelerated the spread of drug-resistant bacteria and led to the emergence of multi-drug-resistant "super bacteria", making current antimicrobial drug treatment ineffective. The frequent occurrence of cellular resistance forces humans to continuously develop antibiotics with new structures and new targets. However, the development of new antibiotics requires a lot of time, manpower and material resources, which poses a major threat to human health and property.

[0003] The evolution of bacteria has derived a variety of resistance mechanisms to resist the effects of antibiotics, including reduced drug uptake, changes in drug targets, drug inactivation, and activation of drug efflux pumps, which make traditional antibiotics lose their original antibacterial efficiency and lead to treatment failure. Among them, the formation of biofilms is one of the main mechanisms for bacteria to resist the effects of antibiotics. Research in the past three decades has shown that bacteria use biofilms and quorum sensing to enhance the defense capabilities of the colony. Biofilms are composed of extracellular polymers secreted by bacteria. Using the characteristics of their structure, they form a natural barrier for bacteria, which can escape the effects of the immune system and reduce the uptake of antibiotics, which makes it difficult for bacteria in the biofilm to be killed easily. On the other hand, bacteria regulate the formation of biofilms and the growth of bacteria through quorum sensing, further enhancing the ability of bacteria to cope with harsh environments. At present, there are no effective drugs for this type of resistance mechanism in clinical practice. Compared with traditional antibiotics, biofilm and quorum sensing inhibitors can effectively weaken the resistance of bacteria, can more widely enhance the killing ability of existing antibiotics against resistant bacteria, and because they do not threaten the survival of bacteria, they are not easy to develop resistance. Therefore, the study of new anti-biofilm and quorum sensing drugs is of great significance for overcoming antibiotic resistance.

[0004] The benzothiazole skeleton has good biofilm inhibition activity. Therefore, the present invention, starting from the benzothiazole structure, invents a new class of highly efficient and stable benzothiazole-coupled hydroxamic acid compounds. Such compounds or their pharmaceutically acceptable salts can be used as potential anti-biofilm drugs and antibiotic synergists to enhance the clinical antibacterial treatment effect. Summary of the invention

[0005] In order to overcome the above-mentioned shortcomings and deficiencies of the prior art, the present invention provides a class of benzothiazole-coupled hydroxamic acid compounds, which have a stable chemical structure and have been shown by activity experiments to have anti-biofilm and quorum sensing effects, and have the potential to be new antibiotic synergists.

[0006] In one aspect, the present invention provides a benzothiazole-coupled hydroxamic acid compound represented by formula (I) or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof:

[0007]

[0008] Wherein R1 is a substituent group

[0009] R1=-H, 6′-CH3, 6′-OCH3, 6′-OCH3CH3, 6′-F, 6′-Cl, 6′-Br, 6′-Br, 6′-CF3, 5′-F, 5′-Cl, 5′-CH3, 5′-CF3, 5′-OCH3, 4′-OCH3

[0010] L is one of the following structures:

[0011]

[0012] Preferably, the compound is selected from:

[0013] Table 1 Structure and nomenclature of compounds

[0014]

[0015]

[0016]

[0017]

[0018] The present invention also provides a method for preparing the benzothiazole-coupled hydroxamic acid compound, comprising the following steps:

[0019] (1.1) reacting differently substituted 2-aminobenzothiazole, di-tert-butyl dicarbonate and triethylamine in a solvent, and purifying the reaction solution obtained after the reaction to obtain compounds 1a-1m;

[0020] (1.2) Compound 1a-1m, ethyl bromoacetate and sodium hydride are reacted in a solvent at 0°C, and the resulting reaction solution is purified to obtain compound 2a-2m;

[0021] (1.3) Compounds 2a-2m and sodium hydroxide aqueous solution were reacted in a solvent at 50°C, and the obtained reaction solution was purified to obtain compounds 3a-3m;

[0022] (1.4) Compound 3a-3m, aliphatic amine or aromatic amine methyl ester react with HATU and triethylamine in a solvent, and the obtained reaction solution is purified to obtain an intermediate, and then the intermediate is reacted in a dichloromethane solution of trifluoroacetic acid, and the obtained reaction solution is purified to obtain compound 4a-5a;

[0023] (1.5) Compounds 4a-5a, hydroxylamine hydrochloride and potassium hydroxide are placed in methanol, and the resulting reaction solution is purified to obtain compounds JH1-JH27;

[0024] The synthesis route is:

[0025]

[0026] Preferably, the reaction conditions are: (a) (Boc) 2 O, Et 3 N, DMAP, DCM, rt, 4 h; (b) Ethylbromoacetate, NaH, dry DMF, rt, 3 h; (c) NaOH, H 2 O, MeOH, 40° C., 6 h; (d) HAUT, Et 3 N, dry DCM, 40° C., 4 h; (e) CF 3 COOH, DCM, 40° C., 6 h; (f) NH 2 OH . HCl, KOH, MeOH, 0.5h.

[0027] The present invention also provides the use of the benzothiazole-coupled hydroxamic acid compound or its pharmaceutically acceptable salt, isomer, solvate or prodrug in the preparation of anti-biofilm drugs and antibiotic synergists.

[0028] Preferably, the anti-biofilm drug comprises an anti-biofilm drug for Pseudomonas aeruginosa.

[0029] Preferably, the antibiotic potentiator comprises a ciprofloxacin potentiator.

[0030] The present invention also provides a pharmaceutical composition, which comprises the benzothiazole-coupled hydroxamic acid compound or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof.

[0031] Preferably, the pharmaceutical composition further comprises ciprofloxacin.

[0032] Preferably, the weight ratio of the benzothiazole-coupled hydroxamic acid compound or its pharmaceutically acceptable salt, isomer, solvate or prodrug to ciprofloxacin is 0.2-10.0:1, preferably 0.2:1, 0.3:1, 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, 1.0:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2.0:1, and more preferably 0.83:1.

[0033] Preferably, the medicine or pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers or excipients.

[0034] Beneficial effects:

[0035] Starting from the biologically active benzothiazole structure, this study successfully synthesized a series of chemically stable benzothiazole-coupled hydroxamic acid compounds through structural optimization. The anti-biofilm inhibitory activity of these compounds against Pseudomonas aeruginosa was detected by crystal violet staining, and their antibiotic synergistic effect was evaluated in animal models. The experimental results showed that most compounds showed significant inhibitory effects on the biofilm formation of Pseudomonas aeruginosa and could effectively enhance the antibacterial activity of ciprofloxacin. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 The compound JH2 has a synergistic effect on CIP. (A) Bacterial plating results of the wound infection area of ​​mice in the control group, CIP alone group and JH2-CIP combined group. Each experiment was repeated 3 times; (B) Monitoring of the wound area of ​​mice, and calculating the wound area using Image J; (C) Photographing and monitoring of the wound infection area of ​​mice; (D) Bacterial survival rate in the wound infection area of ​​mice. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. DETAILED DESCRIPTION

[0037] Unless otherwise specified, the reagents used in the examples can be purchased from the market.

[0038] Example 1: Preparation of methyl 5-(2-(benzo[d]thiazol-2-ylamino)acetylamino)pentanoate (Intermediate 4a)

[0039] Step 1: Preparation of tert-butyl benzo[d]thiazol-2-ylcarbamate (Intermediate 1a)

[0040]

[0041] 2-Aminobenzothiazole (0.150 g, 0.001 mol) was dissolved in dichloromethane, triethylamine (0.211 ml, 0.0015 mol) and DMAP (0.012 g, 0.0001 mol) were added, and the mixture was stirred at room temperature. (Boc)2O (0.275 ml, 0.0012 mol) was then added dropwise. After the solvent was added dropwise, the reaction was continued for about 4 hours, and the reaction was terminated. The residual organic solvent in the reaction system was removed, and then the mixture was recrystallized with petroleum ether / ethyl acetate (200 / 20 v / v), and filtered under reduced pressure to obtain 0.195 g of intermediate 1a as an off-white solid with a yield of 78.04%. 1 H NMR (400MHz, DMSO-d6) δ11.74(s,1H),7.92(d,J=7.9Hz,1H),7.70–7.63(m,1H),7.39(t,J=7.7Hz,1H),7.28–7.22(m,1H),1.51(d,J=2.1Hz,9H). 13 CNMR (101 MHz, DMSO) δ 160.05, 153.41, 149.53, 132.56, 126.46, 123.61, 121.89, 120.61, 82.28, 28.31. 1b-1m The preparation method is the same as 1a.

[0042] Step 2: Preparation of N-(benzo[d]thiazol-2-yl)-N-(tert-butoxycarbonyl)glycine ethyl ester (Intermediate 2a)

[0043]

[0044] Intermediate 1a (0.250 g, 0.0001 mol) was placed in a 50 ml two-necked flask, dissolved with an appropriate amount of anhydrous DMF, and placed at 0 ° C. Sodium hydride (0.048 g, 0.0015 mol) was added and stirred for 30 min, and then ethyl bromoacetate (0.167 ml, 0.0015 mol) was added dropwise. After continuing the reaction for about 2 h, a small amount of ice water was added to quench the reaction. After extraction with 100 ml × 3 ethyl acetate, the organic phases were combined, and then washed with 100 ml × 3 saturated NH4Cl liquid and 100 ml × 3 saturated NaCl liquid until the outflowing water layer was neutral. After being fully dried with MgSO4, the mixture was filtered and the solvent was removed. The residue was purified by flash chromatography (petroleum ether-ethyl acetate = 200 / 20 v / v) to obtain intermediate 2a as a white solid 0.201 g, with a yield of 60.01%. 1HNMR(400MHz,Chloroform-d)δ7.77(dd,J=8.3,5.0Hz,2H),7.40(s,1H),7.32–7.26(m,1H),4. 99(d,J=2.8Hz,2H),4.26(dq,J=6.9,4.3,2.8Hz,2H),1.60(s,9H),1.30(dd,J=4.5,2.7Hz,3H). 13 CNMR (101MHz, CDCl3)δ169.12,160.52,148.80,133.67,133.41,125.78,123.51,121.20,120.82,77.24,52.31,48.10,28.08.2b-2m The preparation method is the same as 2a.

[0045] Step 3: Preparation of N-(Benzo[d]thiazol-2-yl)-N-(tert-butyloxycarbonyl)glycine (Intermediate 3a)

[0046]

[0047] The intermediate 2a (0.336 g, 0.001 mol) was dissolved in methanol, and a 2N sodium hydroxide aqueous solution (containing 0.002 mol of sodium hydroxide) was added, and the mixture was stirred at 50°C for 4 h to terminate the reaction. The methanol was removed, and the pH was adjusted to 5 with 1 mol / L HCl. Solids precipitated, and the solids were filtered off under reduced pressure and dried in vacuum to obtain 0.219 g of the intermediate 3a as a white solid, with a yield of 71.11%. 1 H NMR (400MHz, DMSO-d6) δ7.89(d,J=7.9Hz,1H),7.68(d,J=8.1Hz,1H),7.37(t,J=7.7Hz,1H),7.24(t,J=7.6Hz,1H),4.49(s,2H),1.49(s,9H). 13 C NMR (101 MHz, DMSO) δ 169.95, 169.09, 160.79, 148.80, 133.32, 126.59, 124.18, 121.91, 121.23, 82.92, 52.75, 48.33, 28.03. The preparation method of 3b-3m is the same as 3a.

[0048] Step 4: Preparation of methyl 5-(2-(benzo[d]thiazol-2-ylamino)acetylamino)pentanoate (Intermediate 4a)

[0049]

[0050] Dissolve the intermediate 3a (0.308 g, 0.001 mol) in DMF, add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 0.571 g, 0.0015 mol) and triethylamine (0.277 ml, 0.002 mol), stir at room temperature for 30 min, then add 5-aminopentanoic acid methyl ester hydrochloride (0.251 g, 0.0015 mol) for 6 h to terminate the reaction. Extract with 30 ml × 3 ethyl acetate, wash with 30 ml × 3 saturated NaHCO3 solution, 30 ml × 3 saturated NH4Cl solution and 100 ml × 3 saturated NaCl solution respectively, until the outflowing water layer is neutral. Combine the organic phases and dry them thoroughly with MgSO4, then filter and remove the solvent to obtain an oily liquid. Then the oily liquid was placed in a 2 mol trifluoroacetic acid dichloromethane solution, stirred at room temperature for 30 min, and the reaction was terminated. The pH was adjusted to 7 with 1 mol / L NaOH, and the organic phases were combined after extraction with 100 ml × 3 ethyl acetate, followed by washing with 100 ml × 3 saturated NH4Cl and 100 ml × 3 saturated NaCl until the outflowing water layer was neutral. After being fully dried with MgSO4, the mixture was filtered and the solvent was removed. The residue was purified by flash chromatography (petroleum ether-ethyl acetate = 40 / 80 v / v) to obtain intermediate 4a as a white solid 0.201 g yield: 62.51%. 1 H NMR (400MHz, DMSO-d6) δ8.24(d,J=6.0Hz,1H),8.02(d,J=6.0Hz,1H),7.66(dd ,J=7.9,1.9Hz,1H),7.37(dd,J=8.1,1.9Hz,1H),7.28–7.15(m,1H),7.02(dt,J =9.3,4.6Hz,1H),4.08–3.90(m,2H),3.56(d,J=2.0Hz,3H),3.09(q,J=6.3,5.6 Hz,2H),2.29(td,J=7.4,2.0Hz,2H),1.60–1.47(m,2H),1.41(p,J=7.1Hz,2H). 13 C NMR (101 MHz, DMSO) δ 173.73, 169.00, 166.67, 152.63, 131.10, 125.93, 121.53, 121.39, 118.56, 51.64, 47.06, 38.54, 33.36, 28.93, 22.24. The preparation method of 4b-5a is the same as 4a.

[0051] Example 2: N-Hydroxy-4-(2-(Benzo[d]thiazol-2-yl)amino)acetamido)butyramide (Compound JH1)

[0052]

[0053] Hydroxylamine hydrochloride (1.389 g, 0.02 mol) and potassium hydroxide (1.28 g, 0.024 mol) were reacted in methanol solvent for 30 min, and the filtrate was filtered and the reactant 4a (0.161 g, 0.0005 mol) was added to continue the reaction for 30 min. After the reaction was completed, the pH of the reaction solution was adjusted to 7 with dilute hydrochloric acid, and ice water was added to the reaction solution to precipitate solids. If no solids were precipitated, the reaction solution was concentrated under reduced pressure until solids were precipitated. The precipitated solids were filtered and washed with water. The solids were stirred overnight with methanol-ethyl acetate = 1 / 20 v / v, and the product JH1 was filtered to obtain 0.105 g of white solids, with a yield of 65.22%. 1 HNMR (400MHz, DMSO-d6) δ10.33(s,1H),8.67(s,1H),8.22(d,J=6.0Hz,1H),8.01(d,J=6.2Hz,1H),7.67(d,J=7.8Hz,1H),7.38(d,J=8.0 Hz,1H),7.22(t,J=7.9Hz,1H),7.03(t,J=7.7Hz,1H),3.99(d,J=5.3Hz,2H),3.08(q,J=6.7Hz,2H),1.94(t,J=7.4Hz,2H),1.44(m,4H). 13 C NMR (101MHz, DMSO) δ169.43,168.92,166.68,152.65,131.10,125.95,121.52,121.40,118.58,47.05,38.77,32.39,29.14,23.03.HRMS(ESI) of compound JH1:calcd.for C 14 H 19 N4O3S[M+H] + =323.1173, found [M + H] + =323.1165.

[0054] Example 3: N-Hydroxy-5-(2-((6-methoxybenzo[d]thiazol-2-yl)amino)acetamido)pentanamide (Compound JH2)

[0055] Using 4b (0.176 g, 0.0005 mol) and hydroxylamine hydrochloride (1.389 g, 0.02 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH2 as a white solid (0.141 g), with a yield of 80.57%. 1HNMR (400MHz, DMSO-d6) δ10.34(t,J=3.6Hz,1H),8.75–8.57(m,1H),8.00(t,J=6.8Hz,2H),7.43–7.23(m,2H),6.82(t, J=7.1Hz,1H),3.93(d,J=6.1Hz,2H),3.73(t,J=4.0Hz,3H),3.08(q,J=6.3Hz,2H),1.94(q,J=6.9Hz,2H),1.43(m,4H). 13 C NMR(101MHz,DMSO)δ169.46,168.96,165.23,154.94,146.10,131.83,118.77,113.48,106.07,56.01,47.11,38.75,32.38,29.11,23.03.HRMS(ESI)of compoundJH2:calcd.for C 15 H 21 N4O4S[M+H] + =353.1279, found [M + H] + =353.1275.

[0056] Example 4: N-Hydroxy-5-(2-((5-methoxybenzo[d]thiazol-2-yl)amino)acetamido)pentanamide (Compound JH3)

[0057] Using 4c (0.175 g, 0.0005 mol) and hydroxylamine hydrochloride (1.389 g, 0.02 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH3 as an off-white solid (0.113 g), with a yield of 64.57%. 1 HNMR (400MHz, DMSO-d6) δ10.35(d,J=4.4Hz,1H),8.68(s,1H),8.20(t,J=5.4Hz,1H),8.00(q,J=5.4Hz,1H),7.52(dd,J=8.7,4.5Hz,1H),6.98(d, J=4.0Hz,1H),6.65(t,J=8.2,3.6Hz,1H),3.97(t,J=5.3Hz,2H),3.75(d ,J=4.7Hz,3H),3.17–2.95(m,2H),2.08–1.76(m,2H),1.68–1.24(m,4H). 13C NMR(101MHz,DMSO)δ169.43,168.93,165.42,150.50,141.92,132.11,122.24,1 13.71,108.51,56.02,47.06,38.78,32.38,29.15,23.05.HRMS(ESI)ofcompound JH3:calcd.forC 15 H 21 N4O4S[M+H] + =353.1279, found [M + H] + =353.1276.

[0058] Example 5: N-Hydroxy-5-(2-((4-methoxybenzo[d]thiazol-2-yl)amino)acetamido)pentanamide (Compound JH4)

[0059] Using 4d (0.175 g, 0.0005 mol) and hydroxylamine hydrochloride (1.389 g, 0.02 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH4 as a white solid (0.146 g), with a yield of 83.43%. 1 HNMR (400MHz, DMSO-d6) δ10.33(s,1H),8.66(s,1H),8.11(d,J=6.1Hz,1H),8.02(d,J=6.3Hz,1H),7.26(d,J=7.9Hz,1H),6.99(t,J =8.1Hz,1H),6.84(d,J=8.0Hz,1H),4.00(d,J=5.6Hz,2H),3.84(s,3H),3.08(q,J=6.8Hz,2H),1.94(d,J=7.6Hz,2H),1.44(m,4H). 13 C NMR(101MHz,DMSO)δ169.41,168.92,165.42,150.50,141.93,132.12,122.23,1 13.72,108.52,56.02,47.06,38.78,32.38,29.16,23.05.HRMS(ESI)ofcompound JH4:calcd.for C 15 H 21 N4O4S[M+H] + =353.1279, found [M + H] + =353.1277.

[0060] Example 6: N-Hydroxy-5-(2-((6-methylbenzo[d]thiazol-2-yl)amino)acetamido)pentanamide (Compound JH5)

[0061] Using 4e (0.175 g, 0.0005 mol) and hydroxylamine hydrochloride (1.389 g, 0.02 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH5 ​​as a white solid (0.122 g), with a yield of 69.71%. 1 HNMR (400MHz, DMSO-d6) δ10.33(s,1H),8.66(s,1H),8.10(d,J=5.9Hz,1H),7.99(d,J=6.1Hz,1H),7.47(s,1H),7.26(d,J=8.1Hz,1H),7.03(d ,J=8.2Hz,1H),3.96(d,J=5.6Hz,2H),3.07(q,J=6.6Hz,2H),2.32(s,3H),1.94(t,J=7.3Hz,2H),1.48(d,J=7.7Hz,2H),1.39(d,J=7.6Hz,2H). 13 C NMR(101MHz,DMSO)δ169.42,168.99,165.98,150.53,131.18,130.61,126.97,1 21.34,118.26,47.05,38.75,32.39,29.14,23.03,21.23.HRMS(ESI)ofcompound JH5:calcd.forC 15 H 21 N4O3S[M+H] + =337.1329, found [M + H] + =337.1324.

[0062] Example 7: N-Hydroxy-4-(2-((6-methoxybenzo[d]thiazol-2-yl)amino)acetamido)butanamide (Compound JH6)

[0063] Using 4f (0.085 g, 0.00025 mol) and hydroxylamine hydrochloride (1.036 g, 0.015 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH6 as a white solid (0.042 g), with a yield of 49.41%. 1HNMR(400MHz,DMSO-d6)δ10.36(s,1H),8.72(s,1H),8.20–7.89(m,2H),7.44–7.18(m,2H),6.82(d,J=8.7Hz, 1H), 3.95 (d, J = 5.2Hz, 2H), 3.73 (s, 3H), 3.08 (q, J = 6.5Hz, 2H), 1.97 (t, J = 7.3Hz, 2H), 1.63 (p, J = 7.2Hz, 2H). 13 CNMR(101MHz,DMSO)δ169.27,169.19,165.09,154.87,146.71,132.13,118.93,113.40,105.99,55.99,47.08,38.67,30.29,25.83.HRMS(ESI)of compound JH6:calcd.for C 14 H 19 N4O4S[M+H] + =339.1122, found [M + H] + =339.1132.

[0064] Example 8: 5-(2-((5-fluorobenzo[d]thiazol-2-yl)amino)acetamido)-N-hydroxypentanamide (Compound JH7)

[0065] Using 4 g (0.170 g, 0.0005 mol) and hydroxylamine hydrochloride (1.389 g, 0.02 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH7 as a white solid (0.144 g), with a yield of 84.72%. 1 HNMR (400MHz, DMSO-d6) δ10.34(s,1H),8.80(s,1H),8.46(d,J=6.0Hz,1H),8.04(s,1H),7.69(d,J=2.6Hz,1H),7.39(d,J=3.2Hz,1 H),7.05(d,J=8.3Hz,1H),4.00(t,J=3.8Hz,2H),3.07(d,J=6.7Hz,2H),1.95(s,2H),1.49(t,J=7.6Hz,2H),1.39(d,J=7.6Hz,2H). 13 C NMR(101MHz,DMSO)δ169.42,168.89,168.71,161.70(d, 1 J C-F =238.0Hz),153.89(d, 3 J C-F=12.5Hz),126.63,122.21(d, 3 J C-F =10.1Hz),108.72(d, 2 J C-F =24.2Hz),105.16(d, 2 J C-F =24.0Hz),46.99,38.77,32.36,29.11,23.03.HRMS(ESI)ofcompound JH7:calcd.for C 14 H 17 FN4O3SK[M+K] + =379.0637, found[M+K] + =379.0621.

[0066] Example 9: 5-(2-(6-Fluorobenzo[d]thiazol-2-yl)amino)acetamido)-N-hydroxypentanamide (Compound JH8)

[0067] Using 4h (0.170 g, 0.0005 mol) and hydroxylamine hydrochloride (1.389 g, 0.02 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH8 as a white solid (0.144 g), with a yield of 84.72%. 1 HNMR (400MHz, DMSO-d6) δ10.34(s,1H),8.67(s,1H),8.23(t,J=5.8Hz,1H),8.07–7.97(m,1H),7.61(d,J=2.7Hz,1H),7.36(q,J=4.9Hz,1H ),7.05(t,J=9.1Hz,1H),3.97(d,J=5.5Hz,2H),3.07(q,J=6.5Hz,2H),1.94(t,J=7.2Hz,2H),1.49(p,J=7.5Hz,2H),1.38(p,J=7.2Hz,2H). 13 C NMR(101MHz,DMSO)δ169.44,168.85,166.59,157.69(d, 1 J C-F =236.6Hz),149.32,132.15(d, 3 J C-F =11.1Hz),118.99(d, 3 J C-F =8.9Hz),113.25(d, 2 J C-F =23.7Hz),108.31(d, 2J C-F =26.7Hz),47.04,38.77,32.39,29.14,23.03.HRMS(ESI)of compound JH8:calcd.for C 14 H 18 FN4O3S[M+H] + =341.1079, found[M+H] + =341.1072..

[0068] Example 10: 5-(2-(6-(Trifluoromethyl)benzo[d]thiazol-2-yl)amino)acetamido)-N-hydroxypentanamide (Compound JH9)

[0069] Using 4i (0.078 g, 0.0002 mol) and hydroxylamine hydrochloride (0.552 g, 0.008 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH9 as a white solid (0.051 g), with a yield of 65.38%. 1 HNMR (400MHz, DMSO-d6) δ10.34(s,1H),8.78–8.29(m,2H),8.14(s,1H),8.06(q,J=6.9Hz,1H),7.50(t,J=6.6Hz,1H),7.38–7.19 (m,1H),4.01(dd,J=20.3,5.4Hz,2H),3.08(q,J=6.5Hz,2H),1.95(t,J=7.3Hz,2H),1.50(p,J=7.6Hz,2H),1.39(p,J=6.9Hz,2H). 13 C NMR (101MHz, DMSO) δ169.34,168.51,167.26,155.64,140.94,131.79,123.06(q, 3 J C-F =5.7Hz),119.08(q, 3 J C-F =5.9Hz),118.37(q, 4 J C-F =4.7Hz),47.02,38.78,32.36,29.11,23.03.HRMS(ESI)of compound JH9:calcd.forC 15 H 18 F3N4O3S[M+H] + =391.1047, found [M+H] + =391.1046.

[0070] Example 11: 5-(2-((6-chlorobenzo[d]thiazol-2-yl)amino)acetylamino)-N-hydroxypentanamide (Compound JH10)

[0071] Using 4j (0.107 g, 0.0003 mol) and hydroxylamine hydrochloride (0.691 g, 0.01 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH10 as a white solid (0.086 g), with a yield of 80.37%. 1 HNMR (400MHz, DMSO-d6) δ10.31(s,1H),8.25(s,1H),8.02(s,1H),7.67(d,J=7.8Hz,1H),7.38(d,J=7.8Hz,1H),7.22(t,J= 7.9Hz,1H),7.02(t,J=7.0Hz,1H),3.98(s,2H),3.08(t,J=7.1Hz,2H),1.95(q,J=7.6Hz,2H),1.44(dq,J=36.0,7.3Hz,4H). 13 C NMR(101MHz,DMSO)δ169.39,168.93,166.68,152.65,131.11,125.94,121.50,121.39,118.57,47.06,38.76,32.39,29.13,23.04.HRMS(ESI)of compoundJH10:calcd.for C 14 H 18 ClN4O3S[M+H] + =357.0783, found [M + H] + =357.0775.

[0072] Example 12: 5-(2-((5-chlorobenzo[d]thiazol-2-yl)amino)acetamido)-N-hydroxypentanamide (Compound JH11)

[0073] Using 4k (0.071 g, 0.0002 mol) and hydroxylamine hydrochloride (0.691 g, 0.01 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH11 as a white solid (0.046 g), with a yield of 64.79%. 1HNMR (400MHz, DMSO-d6) δ10.34(s,1H),8.69(s,1H),8.41(d,J=5.9Hz,1H),8.04(d,J=6.2Hz,1H),7.67(q,J=3.2Hz,1H),7.18(d,J=10.5Hz ,1H),6.87(t,J=9.2Hz,1H),4.07–3.92(m,2H),3.08(t,J=6.8Hz,2H),1.94(t,J=7.4Hz,2H),1.48(q,J=7.8Hz,2H),1.39(q,J=7.4Hz,2H). 13 C NMR (101MHz, DMSO) δ169.43,168.70,168.40,153.92,130.62,129.92,122.66,121.17,117.93,47.04,38.76,32.37,29.08,23.04.HRMS(ESI) of compound JH11:calcd.for C 14 H 18 ClN4O3S[M+H] + =357.0783, found [M + H] + =357.0765.

[0074] Example 13: 5-(2-((6-bromobenzo[d]thiazol-2-yl)amino)acetamido)-N-hydroxypentanamide (Compound JH12)

[0075] Using 4l (0.201 g, 0.0005 mol) and hydroxylamine hydrochloride (1.389 g, 0.02 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH12 as a white solid (0.144 g), with a yield of 71.64%. 1 HNMR(400MHz,DMSO-d6)δ10.36(s,1H),8.69(s,1H),8.39(s,1H),8.04(s,1H),7.92(s,1H),7.44–7.24(m,2 H), 3.99 (s, 2H), 3.07 (d, J = 6.7Hz, 2H), 1.94 (d, J = 7.3Hz, 2H), 1.49 (t, J = 7.5Hz, 2H), 1.39 (q, J = 7.3Hz, 2H). 13C NMR (101MHz, DMSO) δ169.38,168.70,167.29,151.90,133.35,128.81,123.82,119.95,112.83,47.02,38.77,32.37,29.12,23.04.HRMS(ESI) of compound JH12:calcd.forC 14 H 18 BrN4O3S[M+H] + =401.0278, found [M + H] + =401.0273.

[0076] Example 14: N-Hydroxy-6-(2-((6-methoxybenzo[d]thiazol-2-yl)amino)acetamido)hexanamide (Compound JH13)

[0077] Using 4m (0.088 g, 0.00025 mol) and hydroxylamine hydrochloride (0.691 g, 0.01 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH13 as a white solid (0.055 g), with a yield of 62.50%. 1 H NMR(400MHz,DMSO-d6)δ10.40(s,1H),8.68(s,1H),8.04(dt,J=14.7,5.7Hz ,2H),7.32(d,J=2.6Hz,1H),7.28(d,J=8.7Hz,1H),6.82(dd,J=8.7,2.6Hz,1 H),3.94(d,J=5.5Hz,2H),3.74(s,3H),3.06(q,J=6.6Hz,2H),1.93(t,J=7.4 Hz,2H),1.46(q,J=7.4Hz,2H),1.39(q,J=7.4Hz,2H),1.24(d,J=2.7Hz,2H). 13 C NMR (101MHz, DMSO) δ169.57,169.07,165.11,154.84,146.70,132.13,118.90,113.38,105. 98,56.00,47.11,38.90,32.64,29.27,26.42,25.32.HRMS(ESI)ofcompoundJH13:calcd.for C 16 H 23 N4O4S[M+H] + =367.1435, found [M + H] + =367.1435.

[0078] Example 15: 6-(2-(Benzo[d]thiazol-2-ylamino)acetamido)-N-hydroxyhexanamide (Compound JH14)

[0079] Using 4n (0.084 g, 0.00025 mol) and hydroxylamine hydrochloride (0.691 g, 0.01 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH14 as a white solid (0.060 g), with a yield of 71.43%. 1 HNMR (400MHz, DMSO-d6) δ10.35(s,1H),8.69(s,1H),8.24(s,1H),8.01(s,1H),7.67(d,J=7.6Hz,1H),7.37(d,J=7.9Hz,1H),7.2 2(s,1H),7.02(s,1H),3.98(d,J=5.0Hz,2H),3.07(d,J=6.8Hz,2H),1.92(s,2H),1.44(d,J=29.4Hz,4H),1.24(d,J=7.5Hz,2H). 13 C NMR(101MHz,DMSO)δ169.50,168.88,166.68,152.65,131.11,125.94,121.51,121.40,118.56,47.06,38.93,32.65,29.29,26.43,25.33.HRMS(ESI)of compound JH14:calcd.for C 15 H 21 N4O3S[M+H] + =337.1329, found [M + H] + =337.1322.

[0080] Example 16: 6-(2-((6-chlorobenzo[d]thiazol-2-yl)amino)acetylamino)-N-hydroxyhexanamide (Compound JH15)

[0081] Using 4o (0.074 g, 0.0002 mol) and hydroxylamine hydrochloride (0.691 g, 0.01 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH15 as a white solid (0.066 g), with a yield of 89.19%. 1HNMR(400MHz,DMSO-d6)δ10.34(s,1H),8.67(s,1H),8.37(t,J=5.8Hz,1H), 8.01(t,J=5.6Hz,1H),7.80(d,J=2.3Hz,1H),7.35(d,J=8.5Hz,1H),7.23(d ,J=2.3Hz,1H),3.99(d,J=5.4Hz,2H),3.07(q,J=6.5Hz,2H),1.92(t,J=7.4 Hz,2H),1.48(q,J=7.5Hz,2H),1.39(q,J=7.9Hz,2H),1.24(d,J=5.8Hz,2H). 13 C NMR(101MHz,DMSO)δ169.54,168.71,167.31,151.57,132.83,126.07,125.23,1 21.09,119.41,47.03,38.94,32.65,29.26,26.42,25.32.HRMS(ESI)ofcompound JH15:calcd.for C 15 H 20 ClN4O3S[M+H] + =371.0940, found [M+H] + =371.0937.

[0082] Example 17: 7-(2-(Benzo[d]thiazol-2-ylamino)acetamido)-N-hydroxyheptylamide (Compound JH16)

[0083] Using 4p (0.063 g, 0.00018 mol) and hydroxylamine hydrochloride (0.518 g, 0.0075 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH16 as a light yellow solid (0.048 g), with a yield of 77.16%. 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.66(s,1H),8.23(s,1H),7.98(t,J=5.7Hz,1H),7.71–7.64(m,1H),7.37(d,J=7.9Hz,1H),7.27–7.18( m,1H),7.03(t,J=7.5Hz,1H),3.98(d,J=5.6Hz,2H),3.07(d,J=6.4Hz,2H),1.92(t,J=7.3Hz,2H),1.43(dt,J=24.7,7.0Hz,4H),1.24(m,4H). 13C NMR (101MHz, DMSO) δ169.57,168.90,166.68,152.64,131.10,125.94,121.52,121.40,118.5 5,47.07,38.97,32.68,29.43,28.77,26.52,25.53.HRMS(ESI)ofcompoundJH16:calcd.forC 16 H 23 N4O3S[M+H] + =351.1486, found [M+H] + =351.1482.

[0084] Example 18: 7-(2-((6-chlorobenzo[d]thiazol-2-yl)amino)acetylamino)-N-hydroxyheptylamide (Compound JH17)

[0085] Using 4q (0.115 g, 0.0003 mol) and hydroxylamine hydrochloride (0.828 g, 0.012 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH17 as a white solid (0.098 g), with a yield of 85.22%. 1 HNMR (400MHz, DMSO-d6) δ10.34(s,1H),8.68(s,1H),8.37(t,J=5.6Hz,1H),8.00(t,J=5.6Hz,1H),7.80(s,1H),7.34(d,J=8.5Hz,1H),7. 23(d,J=8.5Hz,1H),3.99(d,J=5.0Hz,2H),3.07(q,J=6.5Hz,2H),1.92(t,J=7.3Hz,2H),1.42(dp,J=20.3,7.0Hz,4H),1.27–1.17(m,4H). 13 C NMR (101MHz, DMSO) δ169.56,168.69,167.29,151.59,132.84,126.07,125.22,121. 09,119.40,47.04,38.98,32.68,29.42,28.76,26.52,25.53.HRMS(ESI)ofcompound JH17:calcd.for C 16 H 22 ClN4O3S[M+H] + =385.1096, found [M + H] + =385.1096.

[0086] Example 19: N-Hydroxy-2-(4-((2-((4-methoxybenzo[d]thiazol-2-yl)amino)acetamido)methyl)phenyl)acetamide (Compound JH18)

[0087] Using 4r (0.120 g, 0.0003 mol) and hydroxylamine hydrochloride (0.828 g, 0.012 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH18 as a light yellow solid (0.107 g), with a yield of 83.33%. 1 H NMR (400MHz, DMSO-d6) δ10.63(s,1H),8.80(s,1H),8.54(d,J=6.5Hz,1H),8.22(d,J=6.0Hz,1H),7.27(d,J=7.8Hz,1H),7.1 9(q,J=7.6Hz,4H),7.00(s,1H),6.86(d,J=8.0Hz,1H),4.29(t,J=3.9Hz,2H),4.15–4.01(m,2H),3.86(s,3H),3.24(s,2H). 13 C NMR(101MHz,DMSO)δ169.29,167.51,165.40,150.58,141.92,137.91,134.91,132.16,12 9.22(2C),127.52(2C),122.31,113.70,108.47,56.04,47.12,42.28,39.63.HRMS(ESI)of compound JH118:calcd.for C 19 H 21 N4O4S[M+H] + =401.1279, found[M+H] + =401.1275.

[0088] Example 20: N-Hydroxy-2-(4-((2-((6-methoxybenzo[d]thiazol-2-yl)amino)acetamido)methyl)phenyl)acetamide (Compound JH19)

[0089] Using 4s (0.081 g, 0.0002 mol) and hydroxylamine hydrochloride (0.552 g, 0.008 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH19 as a white solid (0.064 g), with a yield of 79.01%. 1HNMR (400MHz, DMSO-d6) δ10.62(s,1H),8.79(s,1H),8.48(d,J=7.4Hz,1H),8.09(q,J=4.9Hz,1H),7.31(d,J=12.2Hz,2H),7.26 –7.08(m,4H),6.85(d,J=8.7Hz,1H),4.28(d,J=5.7Hz,2H),4.01(t,J=4.3Hz,2H),3.75(d,J=3.3Hz,3H),3.24(d,J=3.3Hz,2H). 13 C NMR(101MHz,DMSO)δ168.97,167.08,164.64,154.45,146.28,137.56,134.47,131.74,12 8.78(2C),127.06(2C),118.49,113.00,105.57,55.55,46.71,41.80,39.07.HRMS(ESI)of compound JH19:calcd.for C 19 H 21 N4O4S[M+H] + =401.1279, found[M+H] + =401.1273.

[0090] Example 21: 2-((6-ethoxybenzo[d]thiazol-2-yl)amino)-N-(4-(2-(hydroxyamino)-2-oxoethyl)benzyl)acetamide (Compound JH20)

[0091] Using 4t (0.124 g, 0.0003 mol) and hydroxylamine hydrochloride (0.828 g, 0.012 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH20 as a white solid (0.071 g), with a yield of 57.26%. 1 HNMR (400MHz, DMSO-d6) δ10.64(s,1H),8.81(s,1H),8.50(d,J=7.3Hz,1H),8.08(d,J=6.4Hz,1H),7.30(q,J=3.3Hz,2H),7.2 7–7.09(m,4H),6.84(d,J=8.6Hz,1H),4.28(t,J=4.3Hz,2H),4.12–3.89(m,4H),3.24(d,J=2.9Hz,2H),1.31(q,J=6.1Hz,3H). 13C NMR(101MHz,DMSO)δ169.46,167.58,165.10,154.13,146.65,138.00,134.90,132.15,129.23(2C ),127.51(2C),118.96,113.99,106.69,64.01,47.16,42.25,15.22,39.47.HRMS(ESI)ofcompound JH20:calcd.forC 20 H 23 N4O4S[M+H] + =415.1435, found[M+H] + =415.1436.

[0092] Example 22: 2-((5-fluorobenzo[d]thiazol-2-yl)amino)-N-(4-(2-(hydroxyamino)-2-oxoethyl)benzyl)acetamide (Compound JH21)

[0093] Using 4u (0.117 g, 0.0003 mol) and hydroxylamine hydrochloride (0.828 g, 0.012 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH21 as a white solid (0.082 g), with a yield of 70.09%. 1 HNMR (400MHz, DMSO-d6) δ10.62(s,1H),8.79(s,1H),8.50(dt,J=23.3,5.7Hz,2H),7.68(q,J=5.5Hz,1H ),7.19(q,J=7.1Hz,5H),6.98–6.84(m,1H),4.28(d,J=5.8Hz,2H),4.06(d,J=5.6Hz,2H),3.25(s,2H). 13 C NMR(101MHz,DMSO)δ169.02,168.91,167.50,161.72(d, 1 J C-F =238.1Hz),153.90(d, 3 J C-F =12.3Hz),137.93,134.98,129.26(2C),127.56(2C),126.67(d, 4 J C-F =1.6Hz),122.27(d, 3 J C-F =10.2Hz),108.80(d, 2 J C-F =23.8Hz),105.16(d, 2J C-F =24.1Hz),47.07,42.30,40.56.HRMS(ESI)of compoundJH21:calcd.for C 18 H 18 FN4O3S[M+H] + =389.1079, found[M+H] + =389.1073.

[0094] Example 23: N-Hydroxy-2-(4-((2-((6-methylbenzo[d]thiazol-2-yl)amino)acetamido)methyl)phenyl)acetamide (Compound JH22)

[0095] Using 4v (0.096 g, 0.00025 mol) and hydroxylamine hydrochloride (0.691 g, 0.01 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH22 as a yellow solid (0.077 g), with a yield of 80.21%. 1 HNMR (400MHz, DMSO-d6) δ10.62(s,1H),8.79(s,1H),8.51(d,J=6.6Hz,1H),8.20(d,J=6.7Hz,1H),7.48(s,1H),7.33–7.26( m,1H),7.19(q,J=8.1Hz,4H),7.06(d,J=8.2Hz,1H),4.28(t,J=4.1Hz,2H),4.03(t,J=3.8Hz,2H),3.24(s,2H),2.33(s,3H). 13 C NMR (101MHz, DMSO) δ169.33,167.51,165.95,150.53,137.98,134.91,131.22,130.67,12 9.22(2C),127.50(2C),127.00,121.36,118.25,47.13,42.25,40.57,21.23.HRMS(ESI)of compound JH22:calcd.for C 19 H 21 N4O3S[M+H] + =385.1329, found [M + H] + =385.1329.

[0096] Example 24: 2-((5-bromobenzo[d]thiazol-2-yl)amino)-N-(4-(2-(hydroxyamino)-2-oxoethyl)benzyl)acetamide (Compound JH23)

[0097] Using 4w (0.090 g, 0.0002 mol) and hydroxylamine hydrochloride (0.691 g, 0.01 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH23 as a white solid (0.067 g), with a yield of 74.44%. 1 HNMR (400MHz, DMSO-d6) δ10.63(s,1H),8.80(s,1H),8.54(dt,J=11.0,5.8Hz,2H),7.66(d,J=8.3Hz,1H ),7.54(d,J=2.0Hz,1H),7.25–7.14(m,5H),4.28(d,J=5.9Hz,2H),4.07(d,J=5.7Hz,2H),3.25(s,2H). 13 C NMR(101MHz,DMSO)δ168.97,168.20,167.52,154.21,137.91,134.96,130.43,129.26(2C),127.58 (2C),123.99,123.14,120.85,118.74,47.08,42.30,39.98.HRMS(ESI)ofcompoundJH23:calcd.for C 18 H 18 BrN4O3S[M+H] + =449.0278, found [M + H] + =449.0269.

[0098] Example 25: 2-((6-bromobenzo[d]thiazol-2-yl)amino)-N-(4-(2-(hydroxyamino)-2-oxoethyl)benzyl)acetamide (Compound JH24)

[0099] Using 4x (0.135 g, 0.0003 mol) and hydroxylamine hydrochloride (0.828 g, 0.012 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH24 as a white solid (0.067 g), with a yield of 74.44%. 1 HNMR(400MHz,DMSO-d6)δ10.64(s,1H),8.81(s,1H),8.50(d,J=39.0Hz,2H),7.81( s,1H),7.37(s,1H),7.23(d,J=27.0Hz,5H),4.28(s,2H),4.06(s,2H),3.25(s,2H). 13C NMR(101MHz,DMSO)δ169.07,167.54,167.31,151.60,137.95,134.96,132.90,129.25 (2C),127.54(2C),126.12,125.29,121.13,119.44,55.37,47.12,42.29.HRMS(ESI)of compound JH24:calcd.for C 18 H 18 BrN4O3S[M+H] + =449.0278, found [M + H] + =449.0267..

[0100] Example 26: N-Hydroxy-2-(4-(2-((6-methoxybenzo[d]thiazol-2-yl)amino)acetamido)phenyl)acetamide (Compound JH25)

[0101] Using 4y (0.077 g, 0.0002 mol) and hydroxylamine hydrochloride (0.552 g, 0.008 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH25 as a white solid (0.061 g), with a yield of 79.22%. 1 HNMR (400MHz, DMSO-d6) δ10.62(s,1H),10.08(s,1H),8.81(s,1H),8.13(t,J=5.8Hz,1H),7.52(d,J=8.1Hz,2H),7.32(d,J=2.6Hz ,1H),7.28(d,J=8.7Hz,1H),7.19(d,J=8.1Hz,2H),6.81(dd,J=8.8,2.6Hz,1H),4.18(d,J=5.8Hz,2H),3.73(s,3H),3.22(s,2H). 13 C NMR (101MHz, DMSO) δ168.16,167.59,165.17,154.91,146.65,137.79,132.15,131.34 ,129.69(2C),119.46(2C),118.97,113.43,106.03,56.00,47.50,39.23.HRMS(ESI)of compound JH25:calcd.for C 18 H 19 N4O4S[M+H] + =387.1122, found [M + H] + =387.1129.

[0102] Example 27: N-Hydroxy-2-(4-(2-((6-methylbenzo[d]thiazol-2-yl)amino)acetamido)phenyl)acetamide (Compound JH26)

[0103] Using 4z (0.111 g, 0.0003 mol) and hydroxylamine hydrochloride (0.828 g, 0.012 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH26 as a white solid (0.0547 g), with a yield of 49.28%. 1 HNMR (400MHz, DMSO-d6) δ10.64(s,1H),10.10(s,1H),8.83(s,1H),8.25(t,J=5.8Hz,1H),7.54(d,J=8.1Hz,2H),7.48(d,J=1.7Hz ,1H),7.27(d,J=8.1Hz,1H),7.20(d,J=8.1Hz,2H),7.03(dd,J=8.2,1.8Hz,1H),4.21(d,J=5.6Hz,2H),3.24(s,2H),2.32(s,3H). 13 C NMR(101MHz,DMSO)δ168.09,167.58,166.05,150.46,137.79,131.35,131.20,130.72 ,129.70(2C),126.99,121.39,119.46(2C),118.29,47.49,39.24,21.23.HRMS(ESI)of compound JH26:calcd.for C 18 H 19 N4O3S[M+H] + =371.1173, found [M + H] + =371.1174.

[0104] Example 28: N-Hydroxy-2-(4-(2-((4-methoxybenzo[d]thiazol-2-yl)amino)acetamido)phenyl)acetamide (Compound JH27)

[0105] Using 5a (0.112 g, 0.0003 mol) and hydroxylamine hydrochloride (0.828 g, 0.012 mol) as raw materials, the synthesis method was the same as compound JH1 to obtain JH27 as a white solid (0.0479 g), with a yield of 43.15%. 1HNMR (400MHz, DMSO-d6) δ10.63(s,1H),10.15(s,1H),8.82(s,1H),8.27(t,J=5.8Hz,1H),7.53(d,J=8.3Hz,2H),7.27(d,J=7. 8Hz,1H),7.20(d,J=8.2Hz,2H),7.00(t,J=8.0Hz,1H),6.84(d,J=8.0Hz,1H),4.24(d,J=5.7Hz,2H),3.82(s,3H),3.23(s,2H). 13 C NMR(101MHz,DMSO)δ168.04,167.61,165.50,150.50,141.83,137.78,132.12,131.35 ,129.71(2C),122.35,119.45(2C),113.75,108.51,55.99,47.54,39.24.HRMS(ESI)of compound JH27:calcd.for C 18 H 19 N4O4S[M+H] + =387.1122, found [M + H] + =387.1123.

[0106] Example 29: Inhibitory activity of compounds against Pseudomonas aeruginosa biofilm

[0107] Experimental method: This experiment uses crystal violet staining to quantitatively determine the biofilm content: the maximum screening concentration of the compound is 10μM, a blank control group and a negative control group are set up, azithromycin is used as the positive control drug, and the compound is diluted into a series of concentration gradients by double dilution method. 75μL of drug-containing culture medium is added to each 96-well plate, and then 75μL of Pseudomonas aeruginosa PAO1 bacterial solution is inoculated. After culturing in a 37℃ constant temperature incubator for 24h, the planktonic bacteria are washed away with PBS buffer solution, 160μL of methanol is added to fix at room temperature for 20min, 150μL of 0.1% crystal violet stain is added, and the mixture is placed at room temperature for 15min. After washing away the excess dye with distilled water, 150μL of 33% glacial acetic acid is added, and after oscillation, the absorbance value is measured at a wavelength of 570nm using an enzyme reader. The experiment is repeated three times to screen out compounds with biofilm inhibition activity. The results of the biofilm inhibition activity experiment are shown in Table 1.

[0108] Table 1 Inhibition rate of benzothiazole-coupled hydroxamic acid derivatives on Pseudomonas aeruginosa PAO1 biofilm

[0109]

[0110] a, For all compounds, their IC 50 All of them were measured when the biofilm inhibition rate of the positive drug azithromycin was >50%.

[0111] Experimental results: According to the experimental results in Table 1, it can be concluded that with the increase of the length of the fatty carbon chain, the inhibitory activity of the compound against PAO1 biofilm decreased, among which the compound with a carbon chain length of 4 had the best biofilm inhibitory activity; this phenomenon may be related to the effect of the carbon chain length on the solubility of the compound. In addition, when the fatty chain was replaced by an aromatic ring, the biofilm inhibitory activity of the compound did not change significantly. In general, the biofilm inhibitory activity of compounds substituted with electron-donating groups was better than that of compounds substituted with electron-withdrawing groups; methoxy>methyl>electron-withdrawing group; among the methoxy substitutions at different positions, the substituted compound at position 4 showed the strongest biofilm inhibitory activity, followed by positions 6 and 5. This trend may be related to the position of the substituent on the benzene ring, which affects the electron distribution and spatial configuration of the molecule. Among them, compound JH2 substituted with methoxy at position 6 has the best biofilm inhibitory activity, IC 50 =0.40±0.09μM.

[0112] Example 30: Study on the synergistic effect of compound JH2 on ciprofloxacin (CIP) in a mouse wound infection model

[0113] Experimental methods: This animal experiment was conducted in accordance with the relevant national regulations on animal experiments. Pseudomonas aeruginosa PAO1 strain was used in this animal experiment, and female 5-week-old Babl / c mice were purchased from SPF (Beijing) Biotechnology.co. Ltd. During the entire experiment, the living environment of the mice was maintained at a constant temperature of 25°C, with a 12-h light / night cycle, and sufficient food and water were provided. First, 20 mice were randomly divided into 4 groups, 5 mice per group. After anesthesia with 4% chloral hydrate, the hair on the back of the mice was shaved, a 4-5 mm circular wound was established on the back of each mouse, and 5×10 8 CFU of PAO1 was used to establish a wound infection model for 24 hours. Then different groups of mice were treated with different drugs (normal saline, 1 mg / ml CIP, 0.002 mg / ml CIP+2.5 μM JH2, 0.001 mg / ml CIP+2.5 μM JH2). After 3 consecutive days of administration, the skin of the mouse wound was taken for CFU counting, and the mouse wound was photographed and monitored every day to calculate the wound area.

[0114] Experimental results: Figure 1As shown in the figure, after 3 days of continuous administration of the drug to mice, the bacteria in the wound of the mice in the combined administration treatment group were significantly reduced compared with the bacteria in the saline group (Control). By calculating the survival rate of bacteria in the wound, the survival rate of bacteria in the saline group was 100%. After 3 days of treatment with 1mg / ml CIP, the bacteria were basically completely eliminated, and the survival rate was 0%. After the combined treatment of 0.001mg / ml CIP+2.5μM JH2, very few bacteria survived, and the survival rate was close to 0%, indicating that the synergistic effect of JH2 on CIP reached 500-1000 times. Subsequently, the monitoring of the wound area of ​​mice also obtained consistent experimental results. After 3 days of combined administration of JH2 and diluted CIP, the wound area was basically the same as that of the CIP group alone, and both were much smaller than the wound area of ​​the Control group. On the 9th day, the degree of wound healing reached about 75%. The above results all show that JH2, as a biofilm inhibitor, has a good synergistic effect on CIP, and its synergistic effect is 500-1000 times.

[0115] The above embodiments are only used to help understand the method and core idea of ​​the present invention. It should be noted that, for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principles of the present invention, and these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A benzothiazole-coupled hydroxamic acid compound represented by formula (I) or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof: Wherein R1 is a substituent group R1=-H, 6′-CH3, 6′-OCH3, 6′-OCH2CH3, 6′-F, 6′-Cl, 6′-Br, 6′-Br, 6′-CF3, 5′-F, 5′-Cl, 5′-CH3, 5′-CF3, 5′-OCH3, 4′-OCH3 L is one of the following structures:

2. The benzothiazole-coupled hydroxamic acid compound or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof according to claim 1, characterized in that The compound is selected from:

3. A method for preparing the benzothiazole coupled hydroxamic acid compound as claimed in claim 1, characterized in that The following steps are involved: (1.1) reacting differently substituted 2-aminobenzothiazole, di-tert-butyl dicarbonate and triethylamine in a solvent, and purifying the reaction solution obtained after the reaction to obtain compounds 1a-1m; (1.2) Compound 1a-1m, ethyl bromoacetate and sodium hydride are reacted in a solvent at 0°C, and the resulting reaction solution is purified to obtain compound 2a-2m; (1.3) Compounds 2a-2m and sodium hydroxide aqueous solution were reacted in a solvent at 50°C, and the obtained reaction solution was purified to obtain compounds 3a-3m; (1.4) Compound 3a-3m, aliphatic amine or aromatic amine methyl ester react with HATU and triethylamine in a solvent, and the obtained reaction solution is purified to obtain an intermediate, and then the intermediate is reacted in a dichloromethane solution of trifluoroacetic acid, and the obtained reaction solution is purified to obtain compound 4a-5a; (1.5) Compounds 4a-5a, hydroxylamine hydrochloride and potassium hydroxide are placed in methanol, and the resulting reaction solution is purified to obtain compounds JH1-JH27; The synthesis route is:

4. Use of the benzothiazole-coupled hydroxamic acid compound or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof as claimed in claim 1 or 2 in the preparation of anti-biofilm drugs and antibiotic synergists.

5. The use according to claim 5, characterized in that: The anti-biofilm drug includes an anti-biofilm drug for Pseudomonas aeruginosa; the antibiotic enhancer includes a ciprofloxacin enhancer.

6. A pharmaceutical composition comprising the benzothiazole-coupled hydroxamic acid compound according to claim 1 or 2 or a pharmaceutically acceptable salt, isomer, solvate or prodrug thereof.

7. The pharmaceutical composition according to claim 6, characterized in that The pharmaceutical composition also comprises ciprofloxacin.

8. The pharmaceutical composition according to claim 7, characterized in that The weight ratio of the benzothiazole-coupled hydroxamic acid compound or its pharmaceutically acceptable salt, isomer, solvate or prodrug to ciprofloxacin is 0.2-10.0:1, preferably 0.8-2.0:1.

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