CN113801153A - 一种含硼酸及硼酸频哪醇酯基团的苯磺酰胺类hbv衣壳蛋白抑制剂及其制备方法与应用 - Google Patents
一种含硼酸及硼酸频哪醇酯基团的苯磺酰胺类hbv衣壳蛋白抑制剂及其制备方法与应用 Download PDFInfo
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- CN113801153A CN113801153A CN202111188493.2A CN202111188493A CN113801153A CN 113801153 A CN113801153 A CN 113801153A CN 202111188493 A CN202111188493 A CN 202111188493A CN 113801153 A CN113801153 A CN 113801153A
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- boric acid
- boronic acid
- pinacol ester
- hbv
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Classifications
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- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
- C07F5/02—Boron compounds
- C07F5/025—Boronic and borinic acid compounds
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- A—HUMAN NECESSITIES
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Abstract
本发明提供了一种苯磺酰胺‑硼酸级硼酸酯类衍生物及其制备方法和应用。所述化合物具有式I所示的结构。本发明还涉及含有式I结构化合物的制备方法，药物组合物以及提供上述化合物在制备抗HBV药物中的应用。
Description
技术领域
本发明属于医药技术领域，具体涉及一种含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂，本发明还涉及这类抑制剂的制备方法及其作为抗乙型肝炎病毒(HBV)抑制剂在制备抗乙肝药物中的应用。
背景技术
乙肝是由乙型肝炎病毒(HBV)所致的重大传染性疾病，长期发展可导致急慢性病毒性肝炎、肝硬化(liver cirrhosis,LC)、肝代谢失常和肝癌(hepatocellularcarcinoma,HCC)等疾病。目前，乙肝患者的病情虽然在相继上市的抗HBV药物的作用下得到了一定的控制，但现有医药治疗策略仍无法完全治愈慢性乙型肝炎。因此，研究开发高效、低毒和抗耐药的新型HBV抑制剂依然是当前抗乙肝药物研发的热点。基于对HBV生命周期的认识和分子生物学的研究，HBV核衣壳在病毒的生命周期中具有重要作用，其不仅作为HBV基因组的生成场所，也是HBV基因组避免被宿主清除的庇护所，更参与了分泌、包膜及成熟等重要生命周期，因此HBV核衣壳可作为理想的靶点进行药物开发设计。HBV核衣壳抑制剂按结构大致分为丙烯酰胺类抑制剂(Phenylpropenamides，PPA)、苯磺酰胺类抑制剂(Sulfamoybenzamide，SBA)、双噻唑类抑制剂、哒嗪酮类抑制剂、二氢嘧啶类抑制剂、包括酰胺类抑制剂(Benzamide derivatives,BA)在内的其他类抑制剂等。
苯磺酰胺类抑制剂可通过结构优化得到活性较好的衍生物，具有良好的发展前景。目前已经合成的部分化合物对突变株的活性达微摩尔水平，代表化合物有DVR-23和NVR3-778。相比于二氢嘧啶类抑制剂代表药物GLS4，NVR-3-778在体外动物实验中药代动力学性质较好，口服生物利用度达84.6％。然而，活性较GLS4仍有差距。因此，如何在保留苯磺酰胺类结构优势的基础上，设计合成出更高效安全的衣壳蛋白抑制剂这一研究具有重要意义。
发明内容
本发明针对现有技术的不足，提供一种含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂及其制备方法，本发明还提供了上述化合物的部分活性筛选结果及其用途。
本发明的技术方案如下：
1.含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂
本发明所涉及的是一种含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂，或其药学上可接受的盐、酯或前药，具有如下通式I所示的结构：
其中，
A为：三元至六元脂肪环或芳香杂环；
R1为：H、甲基、卤原子、氰基；
R2为：H、甲基、卤原子、氰基；
R3为：H、甲基、卤原子、氰基；
R4为氢原子或者卤原子；
R5为硼酸或硼酸频哪醇酯；
根据本发明优选的，A为苯环，R5如下：
根据本发明进一步优选的，含硼酸及硼酸频哪醇酯基团的苯磺酰胺类化合物是具有下列结构的化合物之一：
表1.目标化合物苯磺酰胺类-硼酸类衍生物的结构
2.含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂的制备方法
苯磺酰胺类-硼酸及硼酸酯类衍生物的制备方法，步骤包括：首先以3-氯磺酰基-4-氟苯甲酸或3-氯磺酰基苯甲酸为起始原料，在甲苯溶液中以二氯亚砜为酰氯化试剂进行羧基的氯酰化反应得到关键中间体I-2；在甲苯溶液中，中间体2与苯胺片段发生亲核酰基取代反应得到重要中间体I-3；将含硼酸及硼酸酯的氨基片段在乙腈中与中间体I-3反应，再加入三乙胺为缚酸剂反应得到终产物I-4。
合成路线如下：
试剂和条件：(Ⅰ)甲苯，二氯亚砜，DMF，回流；(II)苯胺片段，甲苯，回流；(III)含硼酸及硼酸酯的氨基片段，乙腈，三乙胺，产率50％，常温搅拌；
其中，
R1为：H、甲基、卤素原子、氰基；
R2为：H、甲基、卤素原子、氰基；
R3为：H、甲基、卤素原子、氰基；
R5为硼酸或硼酸频哪醇酯。
本发明所述的室温为20-30℃。
根据本发明优选的，含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物的制备方法，具体步骤如下：
(1)称取3-氯磺酰基苯甲酸I-1a或3-氯磺酰基-4-氟苯甲酸I-1b，并滴加数滴N,N-二甲基甲酰胺催化反应，投料完成后120℃下甲苯回流4h；反应结束后，蒸干甲苯和剩余的二氯亚砜得到中间体I-2；中间体I-2不稳定，迅速投至下一步反应；
(2)将中间体I-2溶于甲苯中，80℃下加入苯胺片段，投料完成后120℃回流2h；反应结束后，蒸干甲苯，得到中间体I-3；
(3)将中间体中间体I-3溶于乙腈中，向反应滴加不同含氨基的硼酸和硼酸酯片段的乙腈溶液和三乙胺，室温搅拌12h；反应结束后，萃取，快速柱色谱分离，重结晶得到目标化合物I-4。
3.含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物的应用
本发明公开了含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物的抗HBV活性筛选结果及其应用。通过实验证明本发明的硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物可作为HBV非核苷类抑制剂用于制备抗乙肝药物。
对所合成的目标化合物I-4通过PCR法测定了药物作用下抑制HBV DNA复制活性，通过MTS法测定了化合物的细胞毒性；同时，选择先导化合物NVR3-778和上市药物拉米夫定为阳性对照。对合成的共28个化合物进行了细胞水平的体外抗HBV DNA复制活性初步筛选，其中共18个化合物在20μM和5μM浓度水平对HBV DNA的表达产生了明显的抑制效果。
对这18个化合物进行进一步的体外抗HBV活性评价，通过MTS法测定了药物在不同浓度下的细胞毒性；通过PCR法测定了药物在不同浓度下抑制HBV DNA复制活性。选择先导化合物NVR3-778和上市药物拉米夫定为阳性对照，每个化合物设置五个浓度梯度，分别计算出CC50、EC50和选择性系数SI。
本发明的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物是一类结构新颖的非核苷类HBV抑制剂，可作为抗HBV的先导化合物。
本发明的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物可作为非核苷类HBV抑制剂应用。具体地说，作为HBV抑制剂用来制备抗乙肝药物。
一种抗HBV药物组合物，包括本发明的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物和一种或多种药学上可接受载体或赋形剂。
本发明公开了含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物、其制备方法、抗HBV活性筛选结果及其作为抗HBV抑制剂的首次应用。实验证明本发明的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物可作为HBV抑制剂用于制备抗乙肝药物。
具体实施方式
通过下述实例有助于理解本发明，但是不能限制本发明的内容。
合成路线：
试剂及条件:Ⅰ甲苯，二氯亚砜，DMF，回流；II苯胺片段，甲苯，回流；III苯硼酸及硼酸酯片段，乙腈，三乙胺，产率50％，常温搅拌。
实施例1.化合物I-2的制备
将3-氯磺酰基苯甲酸I-1a或3-氯磺酰基-4-氟苯甲酸I-1b(1.26mmol)溶于30mL甲苯中，室温条件下加入二氯亚砜(5.04mmol)，并加入数滴N,N-二甲基甲酰胺催化反应，120℃回流反应4h；反应结束后减压蒸馏出去溶剂和多余的二氯亚砜，得到黄色油状粗品(I-2)，中间体含酰氯结构不稳定，快速投入下一步反应。
实施例2.化合物I-3的制备
将中间体I-2(1.26mmol)溶于30mL甲苯中，80℃条件下加入苯胺片段(1.26mmol)120℃回流反应2h；反应结束后，冷却至室温，减压蒸馏除去溶剂，得到黄色油状粗品I-3，中间体I-3含不稳定的磺酰氯结构，快速投入下一步反应。
实施例3.化合物I-4的制备
将中间体I-3(1.26mmol)溶于30mL乙腈，室温条件下滴加含氨基的硼酸和硼酸酯片段(1.26mmol)的乙腈溶液和三乙胺(3.78mmol)室温搅拌12h。反应结束，减压除去乙腈，加入水(25mL)，用乙酸乙酯萃取(20mL×3)，收集并合并有机相，用饱和氯化钠萃取一次(25mL)，有机相用无水硫酸镁干燥。过滤，快速柱色谱分离。采用石油醚-乙酸乙酯体系和二氯甲烷-甲醇体系重结晶得到目标化合物I-4。
(3-((3-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)I-4a-1
白色粉末状固体，产率55.6％，熔点182-185℃；1H NMR(400MHz,DMSO-d6)δ10.78(s,1H,NH),10.30(s,1H,NH),8.32(s,1H,PhH),8.15(d,J＝7.8Hz,1H,PhH),8.02(s,2H,BOH),7.94(d,J＝7.9Hz,1H,PhH),7.75–7.69(m,3H,PhH),7.49(s,2H,PhH),7.23–7.13(m,2H,PhH).13C NMR(100MHz,DMSO-d6)δ164.94,δ150.44(d,J＝236.5Hz),140.77,136.98,135.57,135.46,132.16,130.53(d,J＝43.4Hz),130.07,128.62,126.88(d,J＝59.2Hz),122.91,105.24(d,J＝24.5Hz),31.42,26.81,22.94,22.53,14.41.EI-MS:(449.2)[M-H]-,C19H14BF3N2O5S[450.20].
(3-((3-((3,4-二氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4a-2
白色粉末状固体，产率33.2％，熔点159-163℃；1H NMR(400MHz,DMSO-d6)δ10.68(s,1H,NH),10.30(s,1H,NH),8.33(d,J＝1.9Hz,1H,PhH),8.16(d,J＝7.8Hz,1H,PhH),8.03(s,2H,BOH),7.95–7.86(m,2H,PhH),7.77–7.68(m,1H,PhH),7.50–7.45(m,4H,PhH),7.30–7.11(m,2H,PhH).13C NMR(100MHz,DMSO)δ164.68,140.71,δ137.11–135.69(m),131.43(d,J＝142.8Hz),130.04(d,J＝12.1Hz),128.62,126.87(d,J＝53.6Hz),122.86,117.86(d,J＝17.6Hz),117.40,117.33,110.01(d,J＝21.6Hz).,40.63,40.42,40.21,40.00,39.79,39.58,39.37.EI-MS:(431.5)[M-H]-,C19H15BF2N2O5S[432.39].
(4-((3-((3,4-二氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4a-3
白色粉末状固体，产率39.4％，熔点160-162℃；1H NMR(400MHz,DMSO-d6)δ10.67(s,1H,NH),10.53(s,1H,NH),8.36(s,1H,PhH),8.17(d,J＝7.9Hz,1H,PhH),7.96(d,J＝9.9Hz,2H,PhH),7.91(s,2H,BOH),7.73(t,J＝7.9Hz,1H,PhH),7.63(d,J＝7.9Hz,2H,PhH),7.53–7.41(m,2H,PhH),7.07(d,J＝7.9Hz,2H,PhH).13C NMR(100MHz,DMSO)δ164.64,149.39(d,J＝247.3Hz),140.05(d,J＝103.1Hz),135.85,135.68,132.28,130.12,126.63,118.90,117.87(d,J＝17.8Hz),117.38,110.04(d,J＝21.6Hz),40.63,40.42,40.21,40.00,39.79,39.58,39.37.EI-MS:(431.1)[M-H]-,C19H15BF2N2O5S[432.39].
(2-((3-((3,4-二氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4a-4
白色粉末状固体，产率53.4％，熔点140-142℃；1H NMR(400MHz,DMSO-d6)δ10.68(s,1H,NH),10.16(s,1H,NH),8.93(s,2H,BOH),8.32(s,1H,PhH),8.17(d,J＝7.8Hz,1H,PhH),7.91(d,J＝8.7Hz,2H,PhH),7.69(d,J＝8.6Hz,2H,PhH),7.54–7.42(m,2H,PhH),7.33(d,J＝4.0Hz,2H,PhH),7.03(dt,J＝4.7,8.4Hz,1H,PhH).13C NMR(151MHz,DMSO)δ164.49,150.14(d,J＝13.2Hz),148.53(d,J＝12.8Hz),147.06(d,J＝12.3Hz),145.46(d,J＝12.7Hz),143.08,140.00,136.54,136.26(d,J＝8.6Hz),135.80,132.23(d,J＝88.3Hz),130.15(d,J＝25.8Hz),128.28(d,J＝235.2Hz),126.64,123.90,118.96,117.82(d,J＝17.7Hz),117.40(t,J＝4.9Hz),110.07(d,J＝21.6Hz)，40.50,40.36,40.22,40.08,39.94,39.80,39.66.EI-MS:(431.3)[M-H]-,C19H15BF2N2O5S[432.39].
3-(N-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)氨磺酰基)-N-(3,4,5-三氟苯基)苯甲酰胺I-4a-5
白色粉末状固体，产率45.1％，熔点154-156℃；1H NMR(400MHz,DMSO-d6)δ10.77(s,1H,NH),10.69(s,1H,NH),8.35(q,J＝2.2Hz,1H,PhH),8.17(dt,J＝1.4,7.8Hz,1H,PhH),7.99(dt,J＝1.3,8.0Hz,1H,PhH),7.83–7.65(m,3H,PhH),7.56–7.49(m,2H,PhH),7.14(d,J＝8.3Hz,2H,PhH),1.24(s,12H,4CH3).13C NMR(151MHz,DMSO)δ164.85,151.26(d,J＝15.0Hz),149.65(d,J＝5.0Hz),140.81,140.55,136.47,136.09,135.57,134.84,132.80,132.36,131.93,130.47,130.25(d,J＝4.2Hz),126.60,126.56,119.06,105.36(d,J＝5.1Hz),105.23(d,J＝5.1Hz),84.02,65.47,40.50,40.36,40.22,40.08,39.94,39.80,39.66,30.49,25.09,19.10,13.97.EI-MS:(533.2)[M+H]+,C25H24BF3N2O5S[532.34].
(3-((3-((4-氟-3-甲基苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4a-6
白色粉末状固体，产率52.8％，熔点139-141℃；1H NMR(400MHz,DMSO-d6)δ10.45(s,1H,NH),10.29(s,1H,NH),8.32(t,J＝1.9Hz,1H),8.15(d,J＝7.8Hz,1H),8.03(s,2H,BOH),7.93–7.87(m,1H,PhH),7.74–7.62(m,2H,PhH),7.56(ddd,J＝2.7,4.6,7.9Hz,1H,PhH),7.52–7.44(m,2H,PhH),7.24–7.09(m,3H,PhH),2.24(d,J＝2.0Hz,3H,CH3).13C NMR(151MHz,DMSO)δ164.36,157.60(d,J＝240.3Hz),140.74,136.62(d,J＝135.5Hz),135.22,135.20,132.01,130.68,129.84(d,J＝4.9Hz),128.58,127.14,126.56,124.63(d,J＝18.1Hz),124.18(d,J＝4.7Hz),122.85,120.37(d,J＝7.8Hz),115.33(d,J＝23.2Hz),40.49,40.35,40.21,40.07,39.93,39.79,39.65,14.81,14.79.EI-MS:(427.7)[M-H]-,C20H18BFN2O5S[428.24].
(2-((3-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4a-7
白色粉末状固体，产率29.7％，熔点152-154℃；1H NMR(400MHz,DMSO-d6)δ10.77(s,1H,NH),10.15(s,1H,NH),8.90(s,2H,BOH),8.32(d,J＝1.8Hz,1H,PhH),8.20–8.13(m,1H,PhH),7.96–7.90(m,1H,PhH),7.75–7.68(m,4H,PhH),7.39–7.31(m,2H,PhH),7.12–6.99(m,1H,PhH)；13C NMR(100MHz,DMSO)δ165.55,151.24(d,J＝231.6Hz),143.83,140.77,136.80(d,J＝110.0Hz),133.09(d,J＝63.0Hz),132.78,131.15(d,J＝29.8Hz),130.43,129.94,127.44,124.75,106.07(d,J＝24.4Hz),105.95,66.30,41.45,41.24,41.03,40.82,40.61,40.40,40.19,31.29,19.93,14.81.EI-MS:(449.3)[M-H]-,C19H14BF3N2O5S[450.20].
(4-((3-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4a-8
白色粉末状固体，产率23.1％，熔点158-160℃；1H NMR(400MHz,DMSO-d6)δ10.77(s,1H,NH),10.53(s,1H,NH),8.35(t,J＝1.8Hz,1H,PhH),8.16(dt,J＝1.4,7.9Hz,1H,PhH),7.98(dt,J＝1.3,7.9Hz,1H,PhH),7.91(s,2H,BOH),7.77–7.66(m,3H,PhH),7.62(d,J＝8.3Hz,2H,PhH),7.10–7.03(m,2H,PhH).13C NMR(151MHz,DMSO)δ164.91,151.34–151.20(m),149.70,149.65(dd,J＝5.3,10.1Hz),149.60,140.74,139.52,136.47,135.65,135.59,135.55,134.84,132.25,130.23(d,J＝20.8Hz),126.60,119.02,105.38,105.35,105.23(d,J＝5.0Hz),40.50,40.46,40.36,40.22,40.08,39.94,39.80,39.66.EI-MS:(449.4)[M-H]-,C19H14BF3N2O5S[450.20].
(3-(((3-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)甲基)苯基)硼酸I-4a-9
白色粉末状固体，产率44.3％，熔点148-150℃；1H NMR(400MHz,DMSO-d6)δ10.77(s,1H,NH),8.35(d,J＝1.9Hz,1H,NH),8.27(t,J＝6.2Hz,1H,PhH),8.18(dt,J＝1.4,7.8Hz,1H,PhH),8.03(dt,J＝1.4,7.9Hz,1H,PhH),7.98(s,2H,BOH),7.81–7.70(m,3H,PhH),7.70–7.62(m,2H,PhH),7.29–7.19(m,2H,PhH),4.00(d,J＝6.2Hz,2H,CH2).13C NMR(100MHz,DMSO)δ163.88,150.52(d,J＝242.5Hz),136.90,135.77(d,J＝33.0Hz),130.11(d,J＝140.9Hz),127.73,126.05,124.68(d,J＝276.9Hz),123.30,118.38,118.27(d,J＝21.4Hz),105.20(d,J＝24.3Hz),84.26,40.62,40.42,40.21,40.00,39.79,39.58,39.37,25.04.EI-MS:(463.3)[M-H]-,C20H16BF3N2O5S[464.22].
3-(N-(2-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)氨磺酰基)-N-(3,4,5-三氟苯基)苯甲酰胺I-4a-10
白色粉末状固体，产率18.4％，熔点168-170℃；1H NMR(400MHz,DMSO-d6)δ10.80(s,1H,NH),9.02(s,1H,NH),8.25(s,1H,PhH),8.16(d,J＝7.8Hz,1H,PhH),7.81(d,J＝7.9Hz,1H,PhH),7.69(ddd,J＝4.2,6.9,11.4Hz,3H,PhH),7.56–7.49(m,1H,PhH),7.49–7.40(m,1H,PhH),7.32(d,J＝8.2Hz,1H,PhH),7.13(t,J＝7.3Hz,1H,PhH),1.26(s,12H,4CH3).13C NMR(100MHz,DMSO)δ164.79,150.46(d,J＝245.4Hz),142.33,139.80,135.98(d,J＝99.2Hz),132.78(d,J＝50.1Hz),130.28(d,J＝51.9Hz),126.59,125.24,121.61,105.13(d,J＝22.4Hz),84.77,40.63,40.43,40.22,40.12,40.01,39.80,39.59,39.38,24.92.EI-MS:(533.6)[M+H]+,C25H24BF3N2O5S[532.34].
(3-((2-氟-5-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4b-1
白粉末状固体，产率43.8％，熔点148-151℃；1H NMR(400MHz,DMSO-d6)δ10.75(s,1H,NH),10.61(s,1H,NH),8.38(dd,J＝2.3,6.7Hz,1H,PhH),8.24(ddd,J＝2.4,4.6,7.5Hz,1H,PhH),8.01(s,2H,BOH),7.72–7.64(m,2H,PhH),7.62(d,J＝9.1Hz,1H,PhH),7.50(dd,J＝4.6,9.2Hz,2H,PhH),7.21(t,J＝7.6Hz,1H,PhH),7.18–7.12(m,1H,PhH).13C NMR(100MHz,DMSO)δ163.94,150.61(d,J＝243.2Hz),136.38,135.68,131.15,130.79(d,J＝34.1Hz),128.65,128.25,128.17(d,J＝15.2Hz),127.18,122.92,118.16(d,J＝21.7Hz),105.35(d,J＝24.9Hz),40.68,40.47,40.26,40.05,39.85,39.64,39.43,21.49.EI-MS:(467.3)[M-H]-,C19H13BF4N2O5S[468.19].
(4-((2-氟-5-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4b-2
白色粉末状固体，产率55.3％，熔点192-194℃；1H NMR(400MHz,DMSO-d6)δ10.88(s,1H,NH),10.77(s,1H,NH),8.44(dd,J＝2.4,6.8Hz,1H,PhH),8.25(ddd,J＝2.4,4.5,8.7Hz,1H,PhH),7.92(s,2H,BOH),7.69(dd,J＝6.5,10.3Hz,2H,PhH),7.62(dd,J＝4.7,8.7Hz,3H,PhH),7.08(d,J＝8.3Hz,2H,PhH).13C NMR(151MHz,DMSO)δ163.90,160.51(d,J＝260.7Hz),151.36–149.49(m),138.94,136.49,135.80(d,J＝9.4Hz),135.67,135.50,134.86,131.20(d,J＝3.2Hz),130.72,127.97(d,J＝14.2Hz),118.74,118.22(d,J＝21.9Hz),105.39(d,J＝24.3Hz),40.48,40.34,40.20,40.07,39.93,39.79,39.65.EI-MS:(467.6)[M-H]-,C19H13BF4N2O5S[468.19].
4-氟-3-(N-(2-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)氨磺酰基)-N-(3,4,5-三氟苯基)苯甲酰胺I-4b-3
黄色粉末状固体，产率57.4％，熔点>200℃；1H NMR(400MHz,DMSO-d6)δ10.77(s,1H),9.40(s,1H),8.39(d,J＝6.6Hz,1H),8.26(s,1H),7.67(d,J＝16.9Hz,2H),7.64–7.55(m,2H),7.42(t,J＝7.8Hz,1H),7.31(d,J＝8.2Hz,1H),7.13(t,J＝7.4Hz,1H),1.29(s,12H).13C NMR(151MHz,DMSO)δ163.83,160.48(d,J＝260.8Hz),150.44(d,J＝252.8Hz),141.86,136.52,135.96(d,J＝9.7Hz),133.10,131.19,130.66,127.51(d,J＝14.9Hz),125.06,120.48,118.24(d,J＝23.4Hz),105.33(d,J＝21.6Hz),84.86,40.49,40.35,40.21,40.07,39.93,39.79,39.66,24.96.EI-MS:(551.3)[M+H]+,C25H23BF4N2O5S[550.33].
(2-((2-氟-5-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)苯基)硼酸I-4b-4
白色粉末状固体，产率41.6％，熔点137-139℃；1H NMR(400MHz,DMSO-d6)δ10.78(s,1H,NH),10.50(s,1H,NH),9.00(s,2H,BOH),8.46(dd,J＝2.3,6.8Hz,1H,PhH),8.24(ddd,J＝2.3,4.6,8.7Hz,1H,PhH),7.75–7.67(m,3H,PhH),7.62(dd,J＝8.6,10.0Hz,1H,PhH),7.34–7.26(m,2H,PhH),7.02(ddd,J＝3.1,5.2,8.0Hz,1H,PhH).13C NMR(100MHz,DMSO)δ163.84,160.45(d,J＝260.1Hz),149.19(d,J＝10.3Hz),142.56,136.65,136.15(d,J＝10.0Hz),135.52,132.10,131.17(d,J＝3.6Hz),130.87,127.28(d,J＝14.6Hz),123.74,118.28(d,J＝22.0Hz),117.51,105.53–105.11(m),40.62,40.41,40.20,39.99,39.78,39.57,39.37.EI-MS:(467.1)[M-H]-,C19H13BF4N2O5S[468.19].
4-氟-3-(N-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)氨磺酰基)-N-(3,4,5-三氟苯基)苯甲酰胺I-4b-5
白色粉末状固体，产率47.4％，熔点195-198℃；1H NMR(400MHz,DMSO-d6)δ11.05(s,1H,NH),10.78(d,J＝8.0Hz,1H,NH),8.46(dd,J＝2.3,6.8Hz,1H,PhH),8.25(ddd,J＝2.4,4.5,8.6Hz,1H,PhH),7.69(dd,J＝6.5,10.3Hz,2H,PhH),7.63(dd,J＝8.7,9.9Hz,1H,PhH),7.53(d,J＝8.3Hz,2H,PhH),7.15(d,J＝8.4Hz,2H,PhH),1.23(s,12H,4CH3).13C NMR(100MHz,DMSO)δ170.76,163.87,159.18,150.44(d,J＝237.3Hz),140.23,136.13,135.94(d,J＝9.2Hz),131.21,130.77,127.74(d,J＝14.6Hz),118.75,118.26(d,J＝21.7Hz),105.38(d,J＝24.3Hz)84.03,60.19,40.68,40.47,40.27,40.06,39.85,39.64,39.43,25.09,21.20,14.54.EI-MS:(551.4)[M+H]+,C25H23BF4N2O5S[550.33].
(3-((5-((3,4-二氟苯基)氨基甲酰基)-2-氟苯基)磺酰氨基)苯基)硼酸I-4b-6
白色粉末状固体，产率25.6％，熔点145-148℃；1H NMR(400MHz,DMSO-d6)δ10.65(s,1H,NH),10.61(s,1H,NH),8.39(dd,J＝2.4,6.8Hz,1H,PhH),8.25(ddd,J＝2.4,4.6,8.6Hz,1H,PhH),8.02(s,2H,BOH),7.89(ddd,J＝2.5,7.5,13.2Hz,1H,PhH),7.74–7.57(m,1H,PhH),7.54–7.48(m,3H,PhH),7.44(dt,J＝8.9,10.6Hz,1H,PhH),7.26–7.14(m,2H,PhH).13C NMR(100MHz,DMSO)δ163.68,160.41(d,J＝260.2Hz),150.58,150.52(d,J＝10.9Hz),148.10(d,J＝13.3Hz),147.46(d,J＝15.0Hz),145.04(d,J＝12.8Hz),136.41,136.20(d,J＝9.0Hz),135.59(d,J＝9.2Hz),131.43(d,J＝3.3Hz),130.77(d,J＝30.4Hz),128.67,128.04(d,J＝14.7Hz),127.09,122.83,117.96(d,J＝41.5Hz),117.75,117.41,110.10(d,J＝21.5Hz).40.63,40.42,40.21,40.00,39.79,39.59,39.38.EI-MS:(449.5)[M-H]-,C19H14BF3N2O5S[450.20].
(2-((5-((3,4-二氟苯基)氨基甲酰基)-2-氟苯基)磺酰氨基)苯基)硼酸I-4b-7
白色粉末状固体，产率33.4％，熔点142-145℃；1H NMR(400MHz,DMSO-d6)δ10.68(s,1H,NH),10.50(s,1H,NH),9.00(s,2H,BOH),8.47(dd,J＝2.4,6.8Hz,1H,PhH),8.26(ddd,J＝2.4,4.6,8.7Hz,1H,PhH),7.90(ddd,J＝2.5,7.5,13.2Hz,1H,PhH),7.77–7.71(m,1H,PhH),7.60(dd,J＝8.7,10.0Hz,1H,PhH),7.56–7.40(m,2H,PhH),7.36–7.27(m,2H,PhH),7.02(ddd,J＝2.6,5.7,7.9Hz,1H,PhH).13C NMR(100MHz,DMSO)δ163.58,160.33(d,J＝260.1Hz),150.59,148.10(d,J＝13.4Hz),147.41,145.12,142.58,136.65,136.15,136.05,132.11,131.48(d,J＝3.4Hz),130.86,129.76,127.19(d,J＝14.3Hz),123.72,120.43,118.18(d,J＝21.8Hz),117.87(d,J＝17.9Hz),117.49,110.10(d,J＝21.5Hz),40.62,40.42,40.21,40.00,39.79,39.58,39.37.EI-MS:(449.7)[M-H]-,C19H14BF3N2O5S[450.20].
(4-((5-((3,4-二氟苯基)氨基甲酰基)-2-氟苯基)磺酰氨基)苯基)硼酸I-4b-8
白色粉末状固体，产率45.5％，熔点192-194℃；1H NMR(400MHz,DMSO-d6)δ10.87(s,1H),10.68(s,1H),8.45(dd,J＝2.3,6.8Hz,1H),8.26(ddd,J＝2.4,4.6,8.6Hz,1H),7.93(s,2H),7.91–7.85(m,1H),7.62(dd,J＝8.5,10.9Hz,3H),7.50(dd,J＝2.7,6.3Hz,1H),7.48–7.40(m,1H),7.09(d,J＝8.1Hz,2H).13C NMR(100MHz,DMSO)δ163.64,160.38(d,J＝260.2Hz),150.59,150.46,148.10(d,J＝13.3Hz),147.41,145.06(d,J＝12.4Hz),138.97,136.16(d,J＝9.3Hz),135.85,135.70,131.48(d,J＝3.3Hz),130.73,127.80(d,J＝14.5Hz),118.60,118.14(d,J＝21.6Hz),117.85(d,J＝17.8Hz),117.52,117.49,117.46,117.42,110.25,110.04,40.62,40.41,40.20,40.10,39.99,39.78,39.57,39.36.EI-MS:(449.5)[M-H]-,C19H14BF3N2O5S[450.20].
4-氟-3-(N-(3-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)氨磺酰基)-N-(3,4,5-三氟苯基)苯甲酰胺I-4b-9
白色粉末状固体，产率26.2％，熔点180-183℃；1H NMR(400MHz,DMSO-d6)δ10.78(d,J＝1.7Hz,2H),8.42(dd,J＝2.4,6.8Hz,1H),8.25(ddd,J＝2.4,4.5,8.7Hz,1H),7.67(ddd,J＝7.6,10.1,21.2Hz,1H),7.43(t,J＝1.6Hz,3H),7.36–7.29(m,1H),7.28–7.24(m,1H),1.23(s,12H).13C NMR(100MHz,DMSO)δ163.88,150.52(d,J＝242.5Hz),136.90,135.77(d,J＝33.0Hz),130.11(d,J＝140.9Hz),127.73,126.05,124.68(d,J＝276.9Hz),123.30,118.38,118.27(d,J＝21.4Hz),105.20(d,J＝24.3Hz),84.26,40.62,40.42,40.21,40.00,39.79,39.58,39.37,25.04.EI-MS:(551.4)[M+H]+,C25H23BF4N2O5S[550.33].
N-(3,4-二氟苯基)-4-氟-3-(N-(3-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)氨磺酰基)苯甲酰胺I-4b-10
白色粉末状固体，产率38.4％，熔点174-176℃；1H NMR(400MHz,DMSO-d6)δ10.78(s,1H,NH),10.68(s,1H,NH),8.42(dd,J＝2.3,6.8Hz,1H,PhH),8.26(ddd,J＝2.3,4.5,7.3Hz,1H,PhH),7.90(ddd,J＝2.5,7.5,13.2Hz,1H,PhH),7.62(t,J＝9.2Hz,1H,PhH),7.50(t,J＝5.4Hz,1H,PhH),7.43(q,J＝3.9,4.3Hz,2H,PhH),7.33(dd,J＝3.1,5.7Hz,1H,PhH),7.29–7.24(m,2H,PhH),1.23(s,12H,4CH3).13C NMR(100MHz,DMSO)δ163.63,160.38(d,J＝260.2Hz),148.09(d,J＝12.8Hz),147.41(d,J＝12.4Hz),145.00(d,J＝13.4Hz),136.94,136.21,135.79(d,J＝9.3Hz),131.47(d,J＝3.4Hz),130.79,129.41,127.72(d,J＝14.8Hz),126.01,123.27,118.27,118.00(d,J＝9.1Hz),117.78,117.31,109.95(d,J＝21.8Hz),84.26,40.63,40.42,40.21,40.00,39.80,39.59,39.38,25.04.EI-MS:(533)[M+H]+,C25H24BF3N2O5S[532.34].
N-(3,4-二氟苯基)-4-氟-3-(N-(2-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)氨磺酰基)苯甲酰胺I-4b-11
黄色粉末状固体，产率26.2％，熔点>200℃；1H NMR(400MHz,DMSO-d6)δ10.67(s,1H,NH),9.38(s,1H,NH),8.40(d,J＝6.6Hz,1H,PhH),8.27(s,1H,PhH),7.94–7.83(m,1H,PhH),7.60(q,J＝7.1,8.1Hz,2H,PhH),7.52–7.41(m,3H,PhH),7.32(d,J＝8.3Hz,1H,PhH),7.13(t,J＝7.5Hz,1H,PhH),1.29(s,12H,4CH3).
13C NMR(150MHz,DMSO)δ163.57,160.35(d,J＝260.4Hz),150.14(d,J＝13.1Hz),148.52(d,J＝13.4Hz),147.10(d,J＝12.4Hz),145.49(d,J＝11.9Hz),141.93,136.53,136.18(d,J＝8.2Hz),135.91(d,J＝9.5Hz),133.12,131.94,131.52(d,J＝3.0Hz),130.66,127.40(d,J＝14.9Hz),125.01,120.33,118.13(d,J＝22.1Hz),117.83(d,J＝17.9Hz),117.46(t,J＝4.8Hz),110.12(d,J＝21.6Hz),84.88,40.50,40.37,40.23,40.09,39.95,39.81,39.67,24.96.EI-MS:(533.2)[M+H]+,C25H24BF3N2O5S[532.34].
N-(3,4-二氟苯基)-4-氟-3-(N-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苯基)氨磺酰基)苯甲酰胺I-4b-12
黄色粉末状固体，产率56.4％，熔点182-184℃；1H NMR(400MHz,DMSO-d6)δ11.02(s,1H,NH),10.66(s,1H,NH),8.46(d,J＝6.6Hz,1H,PhH),8.26(d,J＝8.0Hz,1H,PhH),7.90(t,J＝10.5Hz,1H,PhH),7.61(t,J＝9.2Hz,1H,PhH),7.53(d,J＝7.9Hz,3H,PhH),7.45(d,J＝9.5Hz,1H,PhH),7.16(d,J＝7.9Hz,2H,PhH),1.24(s,12H,4CH3).13C NMR(100MHz,DMSO)δ163.60,160.34(d,J＝260.3Hz),150.52(d,J＝13.5Hz),148.09(d,J＝15.0Hz),145.06(d,J＝13.6Hz),140.26,136.23,136.16,135.93(d,J＝9.5Hz),131.54,130.77,127.58(d,J＝14.8Hz),118.64,118.29,118.02(dd,J＝20.0,32.7Hz),117.77,117.47,110.14(d,J＝21.7Hz),84.04,40.63,40.42,40.21,40.00,39.80,39.59,39.38,25.10.EI-MS:(533.3)[M+H]+,C25H24BF3N2O5S[532.34].
4-氟-3-(N-(2-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苄基)氨磺酰基)-N-(3,4,5-三氟苯基)苯甲酰胺I-4b-13
白色粉末状固体，产率43.3％，熔点181-183℃；1H NMR(400MHz,DMSO-d6)δ10.75(s,1H,NH),8.38(t,J＝6.3Hz,1H,NH),8.30(dd,J＝2.4,6.8Hz,1H,PhH),8.20(ddd,J＝2.4,4.6,8.7Hz,1H,PhH),7.72(dt,J＝5.1,10.3Hz,2H,PhH),7.59–7.50(m,2H,PhH),7.40–7.32(m,2H,PhH),7.22–7.13(m,1H,PhH),4.41(d,J＝6.2Hz,2H,CH2),1.23(s,12H,4CH3).13C NMR(151MHz,DMSO-d6)δ160.51(d,J＝259.6Hz),150.49(d,J＝237.9Hz),134.92(d,J＝9.8Hz),130.94(d,J＝3.2Hz),129.41(d,J＝14.9Hz),117.95(d,J＝22.3Hz),105.33–105.10(m).13C NMR(151MHz,DMSO)δ164.06,160.51(d,J＝259.6Hz),150.49(d,J＝237.9Hz),143.51,135.77,134.92(d,J＝9.8Hz),131.43,130.94(d,J＝3.2Hz),130.12,129.41(d,J＝14.9Hz),128.20,126.85,117.95(d,J＝22.3Hz),105.33–105.10(m),84.06,45.77,40.50,40.36,40.22,40.08,39.94,39.80,39.67,24.98.EI-MS:(565.2)[M+H]+,C26H25BF4N2O5S[564.36].
4-氟-3-(N-(4-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苄基)氨磺酰基)-N-(3,4,5-三氟苯基)苯甲酰胺I-4b-14
白色粉末状固体，产率55.9％，熔点175-178℃；1H NMR(400MHz,DMSO-d6)δ10.71(s,1H,NH),8.72(t,J＝6.2Hz,1H,NH),8.26(dd,J＝2.3,6.8Hz,1H,PhH),8.19(ddd,J＝2.3,4.5,8.6Hz,1H,PhH),7.71(dt,J＝6.5,13.0Hz,2H,PhH),7.63–7.48(m,3H,PhH),7.22(d,J＝7.7Hz,2H,PhH),4.18(d,J＝6.2Hz,2H,CH2),1.27(s,12H,4CH3).13C NMR(100MHz,DMSO)δ163.93,160.52(d,J＝259.2Hz),150.44(d,J＝235.1Hz),141.00,135.01(d,J＝9.5Hz),134.79,130.36(d,J＝97.1Hz),129.58(d,J＝14.9Hz),127.42,118.01(d,J＝22.3Hz),105.31(d,J＝24.8Hz),84.03,46.45,40.63,40.43,40.22,40.01,39.80,39.59,39.38,25.08.EI-MS:(565.1)[M+H]+,C26H25BF4N2O5S[564.36].
4-氟-3-(N-(3-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)苄基)氨磺酰基)-N-(3,4,5-三氟苯基)苯甲酰胺I-4b-15
白色粉末状固体，产率53.3％，熔点168-170℃；1H NMR(400MHz,DMSO-d6)δ10.71(s,1H,NH),8.69(t,J＝6.3Hz,1H,NH),8.30(dd,J＝2.4,6.8Hz,1H,PhH),8.19(ddd,J＝2.4,4.6,8.7Hz,1H,PhH),7.77–7.62(m,2H,PhH),7.56–7.50(m,2H,PhH),7.50–7.44(m,1H,PhH),7.34(dt,J＝1.6,7.7Hz,1H,PhH),7.23(t,J＝7.5Hz,1H,PhH),4.17(d,J＝6.2Hz,2H,CH2),1.26(s,12H,4CH3).13C NMR(100MHz,DMSO)δ163.97,150.53(d,J＝249.3Hz),137.07,135.99(d,J＝218.0Hz),134.91,133.90(d,J＝39.6Hz),130.99(d,J＝36.6Hz),128.99(d,J＝166.5Hz),117.96(d,J＝22.9Hz),105.21(d,J＝23.7Hz),84.08,46.37,40.63,40.43,40.22,40.01,39.80,39.59,39.38,25.07.EI-MS:(565.4)[M+H]+,C26H25BF4N2O5S[564.36].
3-(((2-氟-5-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)甲基)苯基)硼酸I-4b-16白色粉末状固体，产率37.2％，熔点150-153℃；1H NMR(400MHz,DMSO-d6)δ10.72(s,1H,NH),8.62(t,J＝6.1Hz,1H,NH),8.33(dd,J＝2.3,6.9Hz,1H,PhH),8.21(ddd,J＝2.3,4.4,8.5Hz,1H,PhH),7.94(s,2H,BOH),7.75–7.67(m,2H,PhH),7.66–7.59(m,2H,PhH),7.56(dd,J＝8.7,9.9Hz,1H,PhH),7.27–7.17(m,2H,PhH),4.13(d,J＝6.1Hz,2H,CH2).13C NMR(100MHz,DMSO)δ164.05,163.99,160.52(d,J＝259.2Hz),151.61,150.43(d,J＝239.7Hz),136.46(d,J＝5.8Hz),135.02,133.95(d,J＝19.1Hz),133.37(d,J＝17.1Hz),130.91,129.98,129.83(d,J＝20.6Hz),127.72(d,J＝6.7Hz),118.03(d,J＝22.0Hz),117.84,105.46,105.30(d,J＝18.4Hz),50.61,49.06,46.70,40.63,40.42,40.21,40.00,39.79,39.58,39.38.EI-MS:(481.3)[M-H]-,C20H15BF4N2O5S[482.21].
4-(((2-氟-5-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)甲基)苯基)硼酸I-4b-17白色粉末状固体，产率43.3％，熔点158-160℃；1H NMR(400MHz,DMSO-d6)δ10.74(s,1H,NH),8.67(t,J＝6.2Hz,1H,NH),8.34(dd,J＝2.3,6.8Hz,1H,PhH),8.22(ddd,J＝2.4,4.6,8.7Hz,1H,PhH),7.95(s,2H,BOH),7.71(dd,J＝6.5,10.4Hz,2H,PhH),7.65(d,J＝7.7Hz,2H,PhH),7.62–7.56(m,1H,PhH),7.18(d,J＝7.7Hz,2H,PhH),4.15(d,J＝6.2Hz,2H,CH2).13C NMR(100MHz,DMSO)δ164.06,159.26,150.45(d,J＝243.7Hz),139.79,139.60,135.61(d,J＝3.3Hz),135.60,135.03(d,J＝9.8Hz),134.51,134.31,130.93,129.94,129.58(d,J＝15.1Hz),127.00,126.90,118.04(d,J＝22.3Hz),105.32(d,J＝24.1Hz),50.62,49.06,46.42,40.63,40.42,40.22,40.01,39.80,39.59,39.38.EI-MS:(481.4)[M-H]-,C20H15BF4N2O5S[482.21].
2-(((2-氟-5-((3,4,5-三氟苯基)氨基甲酰基)苯基)磺酰氨基)甲基)苯基)硼酸I-4b-18白色粉末状固体，产率42.4％，熔点183-186℃；1H NMR(400MHz,DMSO-d6)δ10.74(s,1H,NH),8.33(dd,J＝2.3,6.8Hz,1H,NH),8.27(t,J＝6.3Hz,1H,PhH),8.21(ddd,J＝2.4,4.6,8.7Hz,1H,PhH),8.13(s,2H,BOH),7.72(dd,J＝6.5,10.4Hz,2H,PhH),7.56(t,J＝9.3Hz,1H,PhH),7.50–7.46(m,1H,PhH),7.29–7.21(m,2H,PhH),7.14(td,J＝1.7,7.1Hz,1H,PhH),4.34(d,J＝6.2Hz,2H,CH2).13C NMR(151MHz,DMSO)δ164.15,160.53(d,J＝259.2Hz),150.46(d,J＝228.3Hz),141.57,135.63,135.01(d,J＝9.7Hz),134.81,134.33,130.97,129.91,129.65,129.55,129.52,127.73,126.55,117.99(d,J＝22.6Hz),105.33(d,J＝24.5Hz),46.71,43.46,40.48,40.35,40.21,40.07,39.93,39.79,39.65.EI-MS:(481.1)[M-H]-,C20H15BF4N2O5S[482.21].
实施例4.目标化合物的体外抗HBV活性实验(HepDES19细胞)
测试原理：HepDES19细胞是在四环素(可抑制启动子)的控制下，稳定转染了HBV D型基因组基因的HepG2(人肝母细胞瘤)细胞系衍生物。在不存在四环素的情况下，诱导HepDES19细胞HBV复制，添加化合物，并将细胞孵育3天,细胞表达的HBV DNA含量和细胞的生存状况会有所变化。通过定量聚合酶链反应(qPCR)分析HBV DNA含量也就是化合物抑制HBV复制的有效性，得到HBV DNA降低至一半所需要的化合物浓度，即为半数有效浓度(EC50)，表示化合物的抗HBV活性。通过MTS法测试化合物对细胞的毒性大小，得到化合物杀死半数细胞所需浓度，即为半数致死浓度(CC50)，表示化合物的细胞毒性。(Guo H,Jiang D,Zhou T,et al.Journal of virology 2007；81(22):12472-12484；Edwards TC,Lomonosova E,Patel JA,et al.Antiviral research 2017；(143):205-217).
实验方法：
(1)细胞培养。将细胞保存在Dulbecco改良的Eagle培养基(Dulbecco’s modifiedEagle’smedium，DMEM)/F12培养基中，该培养基补充了10％的胎牛血清(FBS)和1％的青霉素/链霉素(P/S)和1μg/mL的四环素。通过从培养基中去除四环素来诱导HBV pgRNA的同步表达。
(2)细胞含药培养。在不存在四环素的情况下，将HepDES19细胞以每孔4x104个细胞的密度接种在96孔板中。诱导HBV复制48小时后，添加最终DMSO浓度为1％的化合物溶液，与细胞共同孵育3天。
(3)细胞活性(EC50)测试方法。细胞在200μL磷酸盐缓冲盐水(PBS)中洗涤，并在150μL核心裂解缓冲液(10mM Tris pH 7.4，1％Tween20，150mM NaCl)中裂解。将细胞在20-23℃的定轨摇床上以350rpm孵育40分钟。将细胞裂解液转移至96孔PCR板中，并以3300×g离心5分钟。将50μL上清液转移至另一个96孔PCR板中，并与20单位微球菌核酸酶和100μMCaCl2混合。将裂解液在37℃孵育1小时，然后将核酸酶在70℃灭活10分钟。将Qiagen蛋白酶(0.005Anson单位)加入到裂解物中，并将混合物孵育过夜，然后在95℃使蛋白酶失活10分钟。
将裂解物用作链优先定量聚合酶链反应(qPCR)分析的模板。用40个循环在95℃下进行15s，在60℃下进行1min进行定量PCR。使用Kappa Probe Force通用PCR预混液。正极性DNA链的引物和探针为5′CATGAACAAGAGATGTGTAGTAGGCAGAG3′，5′GGAGGCTGTAGGCATAAATTGG3′和5′/56-FAM/CTGCGCACC/ZEN/AGCACCATGCA/3IABkFQ。负极性DNA链的引物和探针是5'GCAGATGAGAAGGCACAGA3'，5'CTTCTCCGTCTGCCGTT3'和5'/56-FAM/AGTCCGCGT/ZEN/AAAGAGAGGTGCG/3IABkFQ。使用GraphPad Prism和三参数变量响应对数(抑制剂)-反应算法(variable-response log(inhibitor)-versus-response algorithm)将最低值设置为零，计算正链DNA的EC50值。
(4)细胞毒性(CC50)测试方法。使用CellTiter 96TMAQueous非放射性细胞增殖测定法(MTS)在HepDES19细胞中测量存在化合物时的细胞生存力。在不存在四环素的情况下，将细胞以每孔1x104个细胞的密度接种到96孔板中，两天后使用化合物，并将细胞孵育3天。使用GraphPad Prism和三参数变量响应对数(抑制剂)-反应算法(底值设置为零)计算出50％的细胞毒性浓度(CC50)值。
表2.目标化合物(含硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物，化合物编号和结构式同表1所示)抑制HBV DNA复制及细胞毒性的初步评价
对合成的硼酸及硼酸频哪醇酯类化合物系列共28个化合物进行了细胞水平的体外抗HBV DNA复制活性初步评价，其中共18个化合物在20μM或者5μM浓度水平对HBV DNA的表达产生了明显的抑制效果(HBV DNA表达率低于50％)。对这18个化合物进行复筛，建立浓度梯度，测试计算出其EC50和CC50值如表3所示。
表3.目标化合物抑制HBV DNA复制及细胞毒性的活性评价
| 编号 | 平均EC<sub>50</sub>(μM) | 平均CC<sub>50</sub>(μM) |
| I-4a-1 | 4.85 | 18.1 |
| I-4a-2 | 5.46 | 19.8 |
| I-4a-3 | 8.63 | 14.2 |
| I-4a-4 | 4.12 | 11.5 |
| I-4a-5 | 4.23 | 16.4 |
| I-4a-6 | 3.63 | 15.0 |
| I-4a-7 | 5.14 | 7.6 |
| I-4a-8 | 3.28 | 16.4 |
| I-4a-10 | 4.25 | 8.3 |
| I-4b-6 | 9.13 | 14.9 |
| I-4b-2 | 3.53 | 17.9 |
| I-4b-5 | 3.47 | 17.2 |
| I-4b-10 | 5.07 | 22.8 |
| I-4b-12 | 4.43 | 27.9 |
| I-4b-14 | 1.3 | 27.0 |
| I-4b-15 | 1.6 | 22.4 |
| I-4b-16 | 0.83 | 21.0 |
| I-4b-17 | 1.17 | 28.2 |
| 3TC | 0.40 | >100.00 |
| NVR3-778 | 0.73 | 23.4 |
实验结论分析：新合成的硼酸及硼酸频哪醇酯基团的苯磺酰胺类衍生物呈现出显著的抗HBV活性。经初步的活性筛选，18个化合物抗HBV活性在0.8-9.13μM范围内。化合物I-4b-16活性最好，(EC50＝0.83μM)，与NVR3-778活性相当，具有进一步研究的价值。
4.化合物I-4b-16的水溶性测试化合物的水溶性是评价其成药性的一个重要物理化学性质。依据于中国药典溶解度的测试方法，我们测试了代表性化合物NVR3-778和I-4b-16的溶解度。如表4所示，在不同的pH值下，化合物I-4b-16的水溶性都比NVR3-778有显著的提髙。
表4.化合物NVR3-778和I-4b-16的溶解度
Claims (7)
1.一种含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂，或其药学上可接受的盐、酯或前药，具有如下通式I所示的结构：
其中，
A为：三元至六元脂肪环或芳香杂环；
R1为：H、甲基、卤原子、氰基；
R2为：H、甲基、卤原子、氰基；
R3为：H、甲基、卤原子、氰基；
R4为氢原子或者卤原子；
R5为硼酸或硼酸频哪醇酯。
2.如权利要求1所述的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂，其特征在于，A为苯环，R5如下：
3.如权利要求1或2所述的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂，其特征在于，是具有下列结构的化合物之一：
4.如权利要求1或2所述的如权利要求1所述的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂的制备方法，步骤包括：首先以3-氯磺酰基-4-氟苯甲酸或3-氯磺酰基苯甲酸为起始原料，在甲苯溶液中以二氯亚砜为酰氯化试剂进行羧基的氯酰化反应得到关键中间体I-2；在甲苯溶液中，中间体2与苯胺片段发生亲核酰基取代反应得到重要中间体I-3；将含硼酸及硼酸酯的氨基片段在乙腈中与中间体I-3反应，再加入三乙胺为缚酸剂反应得到终产物I-4；
合成路线如下：
试剂和条件：(Ⅰ)甲苯，二氯亚砜，DMF，回流；(II)苯胺片段，甲苯，回流；(III)含硼酸及硼酸酯的氨基片段，乙腈，三乙胺，产率50％，常温搅拌；
其中，
R1为：H、甲基、卤素原子、氰基；
R2为：H、甲基、卤素原子、氰基；
R3为：H、甲基、卤素原子、氰基；
R5为硼酸或硼酸频哪醇酯。
5.如权利要求4所述的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂的制备方法的制备方法，步骤如下：
(1)称取3-氯磺酰基苯甲酸I-1a或3-氯磺酰基-4-氟苯甲酸I-1b，并滴加数滴N,N-二甲基甲酰胺催化反应，投料完成后120℃下甲苯回流4h；反应结束后，蒸干甲苯和剩余的二氯亚砜得到中间体I-2；中间体I-2不稳定，迅速投至下一步反应；
(2)将中间体I-2溶于甲苯中，80℃下加入苯胺片段，投料完成后120℃回流2h；反应结束后，蒸干甲苯，得到中间体I-3；
(3)将中间体中间体I-3溶于乙腈中，向反应滴加不同含氨基的硼酸和硼酸酯片段的乙腈溶液和三乙胺，室温搅拌12h；反应结束后，萃取，快速柱色谱分离，重结晶得到目标化合物I-4。
6.权利要求1-3任一项所述的含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂在制备抗HBV的药物中的应用。
7.一种抗HBV药物组合物，包含权利要求1-3任一项所述含硼酸及硼酸频哪醇酯基团的苯磺酰胺类HBV衣壳蛋白抑制剂和一种或多种药学上可接受载体或赋形剂。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115677545A (zh) * | 2022-10-28 | 2023-02-03 | 潍坊医学院 | 一种抗hbv磺胺苯甲酰胺类衍生物及其制备方法和应用 |
| CN116410106A (zh) * | 2023-04-13 | 2023-07-11 | 潍坊医学院 | 一种苯甲酰胺类hbv衣壳蛋白抑制剂及其制备方法与应用 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2594554A1 (en) * | 2010-07-13 | 2013-05-22 | Dainippon Sumitomo Pharma Co., Ltd. | Biaryl amide derivative or pharmaceutically acceptable salt thereof |
| CN112912368A (zh) * | 2018-07-19 | 2021-06-04 | 圣拉斐尔医院有限公司 | 乙型肝炎病毒的抑制剂 |
| CN112920208A (zh) * | 2021-01-28 | 2021-06-08 | 山东大学 | 一种含硼酸的吲哚芳基砜类衍生物及其制备方法与应用 |
| CN113072484A (zh) * | 2021-04-12 | 2021-07-06 | 山东大学 | 含有琥珀酸酯的磺胺苯甲酰胺类化合物及其制备方法与应用 |
-
2021
- 2021-10-12 CN CN202111188493.2A patent/CN113801153B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2594554A1 (en) * | 2010-07-13 | 2013-05-22 | Dainippon Sumitomo Pharma Co., Ltd. | Biaryl amide derivative or pharmaceutically acceptable salt thereof |
| CN112912368A (zh) * | 2018-07-19 | 2021-06-04 | 圣拉斐尔医院有限公司 | 乙型肝炎病毒的抑制剂 |
| CN112920208A (zh) * | 2021-01-28 | 2021-06-08 | 山东大学 | 一种含硼酸的吲哚芳基砜类衍生物及其制备方法与应用 |
| CN113072484A (zh) * | 2021-04-12 | 2021-07-06 | 山东大学 | 含有琥珀酸酯的磺胺苯甲酰胺类化合物及其制备方法与应用 |
Non-Patent Citations (3)
| Title |
|---|
| IAN W.WINDSOR等: ""Sub-picomolar Inhibition of HIV‑1 Protease with a Boronic Acid"", 《J.AM.CHEM.SOC.》 * |
| PEK Y.CHONG等: ""Design of N‑Benzoxaborole Benzofuran GSK8175-Optimization of Human Pharmacokinetics Inspired by Metabolites of a Failed Clinical HCV Inhibitor"", 《J.MED.CHEM.》 * |
| SHU SONG等: ""Recent developments in the medicinal chemistry of single boron atom-containing compounds"", 《ACTA PHARMACEUTICA SINICA B》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115677545A (zh) * | 2022-10-28 | 2023-02-03 | 潍坊医学院 | 一种抗hbv磺胺苯甲酰胺类衍生物及其制备方法和应用 |
| CN115677545B (zh) * | 2022-10-28 | 2024-03-15 | 潍坊医学院 | 一种抗hbv磺胺苯甲酰胺类衍生物及其制备方法和应用 |
| CN116410106A (zh) * | 2023-04-13 | 2023-07-11 | 潍坊医学院 | 一种苯甲酰胺类hbv衣壳蛋白抑制剂及其制备方法与应用 |
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