CN110950863A - 一种喹唑啉酮类化合物及其制备方法和应用 - Google Patents
一种喹唑啉酮类化合物及其制备方法和应用 Download PDFInfo
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- CN110950863A CN110950863A CN201911122309.7A CN201911122309A CN110950863A CN 110950863 A CN110950863 A CN 110950863A CN 201911122309 A CN201911122309 A CN 201911122309A CN 110950863 A CN110950863 A CN 110950863A
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- Prior art keywords
- compound
- quinazolinone
- hydrogen
- alkyl
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- -1 Quinazolinone compound Chemical class 0.000 title claims abstract description 33
- 239000001257 hydrogen Substances 0.000 claims abstract description 24
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 24
- 150000002431 hydrogen Chemical class 0.000 claims abstract description 17
- AVRPFRMDMNDIDH-UHFFFAOYSA-N 1h-quinazolin-2-one Chemical compound C1=CC=CC2=NC(O)=NC=C21 AVRPFRMDMNDIDH-UHFFFAOYSA-N 0.000 claims abstract description 15
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 15
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- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 125000003545 alkoxy group Chemical group 0.000 claims abstract description 6
- 125000001475 halogen functional group Chemical group 0.000 claims abstract description 6
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Abstract
本发明公开了一种喹唑啉酮类化合物及其制备方法和应用,所述喹唑啉酮类化合物具有式(Ⅰ)、式(Ⅱ)或式(Ⅲ)所示分子结构;其中,R1为氢、卤基、烷基、烷氧基或硝基;R2为氢、卤基、烷基或烷氧基;R3为氢或烷基;R4为氢或烷基;X1为‑R5‑O‑、‑R5‑NH‑、其中,R5为烷基、烯烃基或烷烃磺酰基。本发明提供的喹唑啉酮类化合物能够与诸多细菌的苏氨酰转运核糖核酸合成酶结合,可作为苏氨酰转运核糖核酸合成酶抑制剂,能够明显地抑制革兰氏阳性(Gram+)和革兰氏阴性(Gram‑)细菌,属于广谱抗菌药物,在治疗细菌感染方面有良好的发展前景。另外,上述喹唑啉酮类化合物以喹唑啉酮类为原料,原料价廉易得,合成方法简单易操作,产率高,产物稳定。
Description
技术领域
本发明涉及医药技术领域,更具体地,涉及一种喹唑啉酮类化合物及其制备方法和应用。
背景技术
抗菌药物广泛用于治疗细菌感染,抗菌药物的滥用导致细菌对抗菌药物产生耐药性。目前,日趋严重的细菌耐药性是临床面临的重大挑战。因此,研发具有新靶标、新作用机制和新骨架的抗菌药物是当前药物研究亟待加强的方向。
氨酰转运核糖核酸合成酶(aminoacyl-tRNA synthetases,aaRSs)是蛋白质翻译过程中的一组关键酶。它们的功能是连接氨基酸与对应的tRNA,合成氨酰tRNA,为核糖体合成蛋白质提供原料。细菌一般含有20种aaRSs,各自催化一种天然氨基酸连接到相应的tRNA上。原则上,抑制任意一种aaRSs的活性,细菌的蛋白质合成都会停止或出错,从而抑制细菌的生长与繁殖。目前,细菌异亮氨酰转运核糖核酸合成酶(isoleucyl-tRNAsynthetase,IleRS)的抑制剂Mupirocin(百多邦)已被WHO列入基本药品目录,被广泛用于治疗包括多种耐药性金黄色葡萄球菌在内的革兰氏阳性球菌引起的皮肤感染。Mupirocin在临床上的成功应用,证实aaRSs家族是抗菌的有效靶标。
苏氨酰转运核糖核酸合成酶(threnoyl-tRNA synthetase,ThrRS)是aaRSs家族中重要的成员组成,由于其关键的催化功能,是一种潜在的抗感染药物靶标。ThrRS虽然在真核和原核细胞中普遍存在,但由于真核生物和原核生物该酶在进化过程中产生了巨大的差异,因此,可以利用这一特点开发出具有特异性抑制细菌酶系的药物。并且不同菌种之间都存在这一酶系,可以研发出广谱抗菌药物。但是,现有技术仍缺乏特异性抑制ThrRS的抗菌药物。
所以,亟需研发出新型抗菌药物,从而应对日趋严重的细菌耐药性。
发明内容
本发明为克服上述现有技术所述的细菌耐药性日趋严重的缺陷,提供一种喹唑啉酮类化合物,提供的喹唑啉酮类化合物可作为ThrRS抑制剂,能够特异性抑制ThrRS,对革兰氏阳性(Gram+)和革兰氏阴性(Gram-)细菌均具有良好抑制效果,属于广谱抗菌药物。
本发明的另一目的在于提供上述喹唑啉酮类化合物的制备方法。
本发明的又一目的在于提供上述喹唑啉酮类化合物在制备抗细菌感染药物中的应用。
为解决上述技术问题,本发明采用的技术方案是:
一种喹唑啉酮类化合物,具有式(Ⅰ)、式(Ⅱ)或式(Ⅲ)所示分子结构;
其中,R1为氢、卤基、烷基、烷氧基或硝基;R2为氢、卤基、烷基或烷氧基;R3为氢或烷基;R4为氢或烷基;
发明人研究发现,上述喹唑啉酮类化合物能够与诸多细菌的ThrRS结合,可作为ThrRS抑制剂,能够明显地抑制革兰氏阳性(Gram+)和革兰氏阴性(Gram-)细菌,属于广谱抗菌药物,例如,大肠杆菌、肠炎沙门杆菌、绿脓杆菌、标准金黄色葡萄球菌、耐甲氧西林金黄色葡萄球菌MRSA或粪肠球菌。所以,上述喹唑啉酮类化合物在治疗细菌感染方面有良好的发展前景。
优选地,R1为氢、氟、氯、溴、甲基、甲氧基或硝基。
优选地,R2为氢、氟、氯、溴、甲基或甲氧基。
优选地,R3为氢、甲基、乙基或异丙基。
优选地,R4为氢或甲基。
优选地,R5为C3~C6烷基、C4烯烃基或烷烃磺酰基。
优选地,R5为-(CH2)3-、-(CH2)4-、-(CH2)5-、-(CH2)6-、(E)-异-2-丁烯基或丙烷磺酰基。
本发明还保护上述喹唑啉酮类化合物的制备方法,所述制备方法为将含喹唑啉酮衍生基团的第一反应物与反应后取代基为X1基团的第二反应物、L-苏氨酸衍生物反应,得到所述喹唑啉酮类化合物。
上述喹唑啉酮类化合物以喹唑啉酮类为原料,原料价廉易得,合成方法简单易操作,产率高,产物稳定。
本发明还保护上述喹唑啉酮类化合物在制备抗细菌感染药物中的应用。
优选地,所述抗细菌感染药物的靶标为细菌型ThrRS。
优选地,所述抗细菌感染药物为抗大肠杆菌、肠炎沙门杆菌、绿脓杆菌、标准金黄色葡萄球菌、耐甲氧西林金黄色葡萄球菌MRSA或粪肠球菌药物。
与现有技术相比,本发明的有益效果是:
本发明提供的喹唑啉酮类化合物能够与诸多细菌的ThrRS结合,可作为ThrRS抑制剂,能够明显地抑制革兰氏阳性(Gram+)和革兰氏阴性(Gram-)细菌,属于广谱抗菌药物,在治疗细菌感染方面有良好的发展前景。
另外,上述喹唑啉酮类化合物以喹唑啉酮类为原料,原料价廉易得,合成方法简单易操作,产率高,产物稳定。
附图说明
图1为化合物30d和35a对肠炎沙门杆菌苏氨酰转运核糖核酸合成酶(Salmonellaenterica threnoyl-tRNA synthetase,SeThrRS)氨酰化的抑制率拟合得到的抑制率和半数抑制浓度IC50值。
图2为化合物30d和35a对SeThrRS的亲和力。
图3为SeThrRS三维结合模式图。
图4为SeThrRS-化合物30d复合物三维结合模式图。
具体实施方式
下面结合具体实施方式对本发明作进一步的说明。
实施例中的原料均可通过市售得到;
除非特别说明,本发明采用的试剂、方法和设备为本技术领域常规试剂、方法和设备。
实施例中各化合物的结构式和编号如表1所示。
表1 化合物结构式
实施例1~5化合物6a-e
实施例1~5的喹唑啉酮类化合物的结构如表1中的6a-e所示。合成路线如下:
(1)化合物3a-e的制备,以化合物3a为例,化合物3a的制备方法如下:
将化合物1(250mg,0.969mmol)与碳酸钾(267mg,1.94mmol)在溶剂DMF中室温搅拌30分钟,随后加入2a(392mg,1.94mmol),反应2小时。经TLC检测,待反应完全后,加水猝灭反应,用乙酸乙酯萃取(3×10.0mL),合并乙酸乙酯相,并用饱和食盐水洗(3×10.0mL),无水硫酸钠干燥,减压除去溶剂,得到的粗品用流动相(环己烷:二氯甲烷=1:2)过硅胶柱纯化,即得化合物3a,其产率约为75%。
(2)化合物5a-e的制备,以化合物5a为例,化合物5a的制备方法如下:
将化合物4(165mg,0.754mmol)、碳酸钾(104mg,0.754mmol)与3a(250mg,0.58mmol),在溶剂DMF中室温搅拌24小时。经TLC检测,待反应完全后,加水猝灭反应,用乙酸乙酯萃取(3×10.0mL),合并乙酸乙酯相,并用饱和食盐水洗(3×10.0mL),无水硫酸钠干燥,减压除去溶剂,得到的粗品用流动相(环己烷:二氯甲烷=1:2)过硅胶柱纯化,即得化合物5a,其产率约为72%。
(3)化合物6a-e的制备,以化合物6a为例,化合物6a的制备方法如下:
将化合物5a(150mg,0.290mmol)溶于1,4-二氧六环(4mL)中,滴加4M盐酸,室温搅拌2小时,经TLC检测,待反应完全后,减压除去溶剂,得到的粗品用流动相(二氯甲烷:甲醇=8:1)过硅胶柱纯化,即得化合物6a,其产率约为91%。
实施例6~9化合物10a-d
实施例6~9的喹唑啉酮类化合物的结构如表1中的10a-d所示。合成路线如下:
(1)化合物8a-d的制备,其制备方法参考实施例1~5中化合物3a的制备方法。
(2)化合物9a-d的制备,以化合物9a为例,化合物9a的制备方法如下:
化合物8a(200mg,0.45mmol)溶解于1,4-二氧六环溶液中,滴加4M盐酸溶液,室温反应过夜。经TLC检测,待反应完全后,减压除去溶剂。将得到的产物(150mg,0.396mmol),4(130mg,0.594mmol)与HATU(226mg,0.594mmol)溶于DMF中,在0℃下,滴加DIPEA(102mg,0.792mmol),在室温下,反应2小时。经TLC检测,待反应完全后,加水猝灭反应,用乙酸乙酯萃取(3×10.0mL),合并乙酸乙酯相,并用饱和食盐水洗(3×10.0mL),无水硫酸钠干燥,减压除去溶剂,得到的粗品用流动相(环己烷:二氯甲烷=1:2)过硅胶柱纯化,即得化合物9a,其产率约为79%。
(3)化合物10a-d的制备,其制备方法参考实施例1~5中化合物6a的制备方法。
实施例10~23化合物30a-n
实施例10~23的喹唑啉酮类化合物的结构如表1中的30a-n所示。合成路线如下:
(1)化合物27a-n的制备,其制备方法参考实施例1~5中化合物3a的制备方法。
(2)化合物28a-n的制备,以化合物28a为例,化合物28a的制备方法如下:
化合物27a(250mg,0.969mmol)、CuI(18mg,0.0969mmol)、叠氮化钠(81.9mg,1.26mmol)在溶剂DMSO(2mL)中室温反应0.5小时。经TLC检测,待反应完全后,加水猝灭反应,用乙酸乙酯萃取(3×10.0mL),合并乙酸乙酯相,并用饱和食盐水洗(3×10.0mL),无水硫酸钠干燥,减压除去溶剂。将得到的中间体(205mg,0.581mmol)和三苯基膦(198mg,0.755mmol)溶解在THF(5.0mL)中,室温搅拌30分钟。在反应液中滴加去离子水(1mL),室温搅拌过夜。减压除去溶剂,将粗品用流动相(二氯甲烷:甲醇=4:1)过硅胶柱纯化,即得化合物28a,其产率约为82%。
(3)化合物29a-n的制备,以化合物29a为例,化合物29a的制备方法如下:
将化合物28a(150mg,0.458mmol)、4(150mg,0.687mmol)与HATU(261mg,0.687mmol)溶解于DMF(5mL)中,在0℃下,滴加DIPEA(118mg,0.916mmol),在室温下,反应2小时。经TLC检测,待反应完全后,加水猝灭反应,用乙酸乙酯萃取(3×10.0mL),合并乙酸乙酯相,并用饱和食盐水洗(3×10.0mL),无水硫酸钠干燥,减压除去溶剂,得到的粗品用流动相(环己烷:二氯甲烷=1:2)过硅胶柱纯化,即得化合物29a,其产率约为86%。
(4)化合物30a-n的制备,其制备方法参考实施例1~5中化合物6a的制备方法。
实施例24~26化合物35a-c
实施例24~26的喹唑啉酮类化合物的结构如表1中的35a-c所示。合成路线如下:
(1)化合物27d、化合物28d的制备,其制备方法参考实施例10~23中化合物28a的制备方法。
(2)化合物32的制备,其制备方法参照实施例10~23中化合物29a的制备方法。
(3)化合物33的制备,制备方法如下:
将化合物32(450mg,0.696mmol)溶于二氯甲烷:哌啶=5:1(10mL),室温反应2小时,经TLC检测,待反应完全后,减压除去溶剂,得到的粗品用流动相(二氯甲烷:甲醇=10:1)过硅胶柱,即得化合物33,其产率约为90%。
(4)化合物35a-c的制备,其制备方法参考实施例1~5中化合物6a的制备方法。
实施例27化合物38
实施例27的喹唑啉酮类化合物的结构如表1中的38所示。合成路线如下:
(1)化合物36的制备,其制备方法参考实施例1~5中化合物3a的制备方法。
(2)化合物37的制备,其制备方法参考实施例10~23中化合物29a的制备方法。
(3)化合物38的制备,其制备方法参考实施例1~5中化合物6a的制备方法。。
制备高纯度SeThrRS蛋白样品
1.沙门肠炎杆菌SeThrRS原核表达质粒的构建
将SeThrRS(UniProKB编号A0A379WUT6)的截短体(242~642)的DNA编码序列插入到pET28a载体,并在该DNA序列的下游插入编码六组氨酸标签的相应DNA序列,构建成表达His6-SeThrRS融合蛋白的原核表达质粒。插入的DNA序列经过DNA测序予以验证。
SeThrRS截短体(242~642)的氨基酸序列为:
MGRDHRKIGKQLDLYHMQEEAPGMVFWHNDGWTIFRELEVFVRSKLKEYQYQEVKGPFMMDRVLWEKTGHWDNYKDAMFTTSSENREYCIKPMNCPGHVQIFNQGLKSYRDLPLRMAEFGSCHRNEPSGALHGLMRVRGFTQDDAHIFCTEEQIRDEVNACIRMVYDMYSTFGFEKIVVKLSTRPDKRIGSDEMWDRAEADLAVALEENNIPFEYQLGEGAFYGPKIEFTLYDCLDRAWQCGTVQLDFSLPSRLSASYVGEDNERKVPVMIHRAILGSMERFIGILTEEFAGFFPTWLAPVQVVVMNITDSQSEYVNELTQKLQNAGIRVKADLRNEKIGFKIREHTLRRVPYMLVCGDKEVEAGKVAVRTRRGKDLGSLDVNDVIEKLQQEIRSRSLQQLEELE。
编码SeThrRS截短体(242~642)的DNA序列为:
ATGGGCCGCGACCATCGTAAAATTGGTAAGCAGCTCGACCTGTATCATATGCAGGAGGAAGCGCCGGGCATGGTGTTCTGGCACAACGACGGCTGGACTATCTTCCGCGAGCTGGAGGTCTTTGTTCGTTCTAAACTCAAAGAGTACCAGTATCAAGAAGTTAAAGGCCCGTTCATGATGGACCGTGTGCTGTGGGAAAAAACCGGGCACTGGGACAACTATAAAGATGCGATGTTCACCACGTCCTCAGAAAACCGCGAATATTGCATCAAGCCGATGAACTGCCCGGGCCACGTTCAGATCTTTAACCAGGGTCTGAAATCCTATCGTGATTTGCCGCTGCGTATGGCGGAATTCGGTAGCTGCCACCGTAACGAGCCATCAGGCGCGCTGCATGGTCTGATGCGCGTACGCGGCTTTACGCAGGATGATGCGCATATCTTCTGCACCGAAGAGCAGATCCGCGATGAAGTTAACGCTTGTATTCGTATGGTCTACGATATGTACAGCACCTTTGGCTTCGAGAAGATCGTCGTCAAGCTTTCCACTCGTCCTGACAAGCGTATCGGCAGCGATGAGATGTGGGATCGTGCTGAGGCGGATCTGGCGGTTGCGCTGGAAGAAAATAATATCCCGTTTGAGTATCAACTGGGTGAAGGCGCATTCTACGGTCCGAAAATTGAATTTACCTTATATGACTGCCTCGATCGTGCATGGCAGTGCGGTACAGTACAGCTGGACTTCTCCTTACCGTCTCGTCTGAGCGCCTCCTATGTAGGCGAAGACAACGAGCGTAAGGTGCCGGTAATGATTCACCGTGCGATTCTTGGGTCGATGGAACGCTTCATCGGTATCCTGACCGAAGAGTTCGCTGGTTTCTTCCCGACATGGCTCGCGCCTGTTCAGGTAGTCGTGATGAATATTACCGATTCGCAGTCTGAATACGTTAACGAATTGACGCAGAAACTACAAAATGCGGGCATTCGTGTAAAAGCAGACTTGAGAAATGAGAAGATTGGCTTTAAAATCCGCGAGCACACTTTACGTCGTGTCCCTTATATGTTGGTCTGTGGTGATAAAGAGGTGGAAGCAGGCAAAGTTGCCGTTCGCACCCGCCGCGGTAAAGACCTGGGCAGCCTGGACGTAAATGACGTGATTGAGAAGCTGCAACAAGAGATTCGCAGCCGCAGTCTTCAACAACTGGAGGAACTCGAG。
2.SeThrRS融合蛋白的表达
将上述His6-SeThrRS质粒转入大肠杆菌BL21(DE3)。将转有质粒的BL21(DE3)细菌在含有100μg/L的氨苄霉素的Luria-Bertani(LB)培养基中37℃温度下220rpm震荡培养,直到OD600=0.6。然后,在培养基中加入0.15mM的诱导剂异丙基硫代半乳糖苷(IPTG),在18℃温度下继续培养,让目标蛋白充分表达。18小时后,离心法收集菌体。
3.SeThrRS的纯化
将上述收集到的菌体用裂解缓冲液(20mM Tris-HCl pH 8.0,500mM NaCl,20mM咪唑)充分悬浮,然后利用超声破碎法在冰浴温度下裂解细胞。高速离心去除细菌裂解液中的细菌残片、细胞器等,收集上清液。利用Ni-NTA亲和层析树脂捕获上清液中的目标蛋白,用裂解缓冲液充分清洗残留在Ni-NTA层析柱上的杂蛋白。在层析柱中加入蛋白酶Ulp1低温过夜孵育,将融合蛋白的His6-蛋白标签与SeThrRS目标蛋白切割开。隔天,收集层析柱穿透峰中的切割后的SeThrRS目标蛋白质样品。利用12%SDS-PAGE检测目标蛋白的纯度。最后,浓缩SeThrRS蛋白样品,得到2mL浓度为30mg/ml的高纯度SeThrRS蛋白样品,储存在-80℃备用。
试验例1
喹唑啉酮化合物6a-e,10a-d,30a-n,35a-c与病原细菌ThrRS结合能力的热稳定性迁移实验(thermal shift assay,TSA)。
在96孔PCR板上配置20μL反应体系(全程在冰上操作),包含:100mM HEPES(pH7.5),150mM NaCl,4×SYPRO orange荧光染料,8μM肠炎沙门杆菌的ThrRS蛋白质(SeThrRS),500μM化合物,轻柔混匀。把96孔PCR板放到StepOnePlus实时荧光定量PCR仪中,在25℃孵育10min,然后按照1℃/min的升温速率从25℃升到95℃。升温期间,每30秒检测一次每孔的荧光信号。随着温度升高,SeThrRS逐渐发生热变性,随之荧光信号增强。以荧光信号作为纵坐标,温度作为横坐标,用origin8软件的玻尔兹曼法拟合绘制蛋白质热变性曲线并推测蛋白质热变性温度Tm。某一个化合物对SeThrRS热稳定性的贡献可以用含有该化合物是蛋白质的Tm值减去不含化合物的空白蛋白质的Tm值,即ΔTm,来评价。ΔTm越大表示结合能力越强。实验测得的喹唑啉酮类化合物对SeThrRS的ΔTm数据见表2。
表2 喹唑啉酮类化合物与SeThrRS的结合能力
注:aNA表示ΔTm值≤0,表示相应化合物在实验条件下不与蛋白结合发生可以测量到的结合。
试验例2
喹唑啉酮类化合物对SeThrRS的抑制活性。
抑制活性测试方法:采用测试ATP消耗量的方法。反应缓冲液(30mM HEPES(pH7.5),150mM NaCl,30mM KCl,40mM MgCl2,1mM DTT和0.1%BSA)用于稀释化合物、蛋白及底物。取5μL 75nM SeThrRS与5μL不同浓度化合物溶液混匀,室温孵育20分钟,加入5μL底物溶液(含12μM ATP,30μM Ile,1.5mg/ml EctRNA)起始反应,室温反应3小时。取10μL反应溶液加到384白底微孔板中,加入10μL试剂(Promega)终止反应,室温放置10分钟测定荧光信号值。实验含A、B和C三组,其中A、B和C分别是不加化合物空白组、加化合物组和不加酶组。抑制率计算采用公式:抑制率%=(RLU(B)–RLU(A))/(RLU(C)–RLU(A))×100。IC50用GraphPad Prism拟合计算。
化合物30d和35a的测试结果如图1所示,均表现出对ThrRS较强的酶抑制活性。
试验例3
喹唑啉酮类化合物对SeThrRS的亲和力测定。
用等温滴定量热仪(VP-ITC MicroCalorimeter,MicroCal Inc.)测定化合物与蛋白质的亲和力。用透析的方法将SeThrRS置换到缓冲溶液20mM HEPES(pH 7.5)和150mMNaCl中,蛋白质浓度为20μM,用同样的缓冲液配制化合物溶液,浓度为200μM。用化合物溶液滴定蛋白质溶液,控制第一滴为5μL,接下来24滴各10μL,每滴间隔180秒。其它参数设定为:Cell Temperature,28℃;Reference Power,5μCal/s;Stirring Speed,394rpm。滴定完成后,用ORIGIN(MicroCal Inc.)软件分析实验数据,拟合曲线,可获得结合常数(Ka),结合化学计量学(N),结合热力学熵变和焓变(ΔS和ΔH)。
化合物30d和35a的测试结果如图2所示,均表现出对ThrRS较强的亲和力。
试验例4
喹唑啉酮类化合物的体外抗菌活性测定。
采用国际标准培养基,最小抑菌浓度(MIC)实验评价喹唑啉酮类化合物6a-e,10a-d,30a-n,35a-c的抗菌活性。测试的菌株有大肠杆菌(E.coli,ATCC25922),粪肠球菌(E.faecalis,ATCC29212),肠炎沙门杆菌(S.enterica,87),标准金黄色葡萄球菌(S.aureus,ATCC29213),耐甲氧西林金黄色葡萄球菌(MRSA,R3708),绿脓杆菌(P.aeruginosa,ATCC27853)。阳性对照用万古霉素与氨苄西林钠。
首先96孔板中第一列加入培养基90μL,其他除空白对照均加入培养基50μL。然后,第一列中加入用DMSO溶解的化合物10μL,混合均匀。吸取第一列中的混合物50μL,置于第二列中混合均匀,再吸取50μL加入第三列。以此类推,最终将化合物6a-e,10a-d,30a-n,35a-c和阳性对照药进行梯度稀释(256、128、64、32、16、8、4、2、1、0.5、0.25μg/mL)。在每孔中分别加入50μL各种菌液。其中最后一列为培养基和菌液的空白对照,最后一排为阳性对照。将96孔板置于37℃,培养12小时。测量600nm处各孔的吸收值,确定化合物对不同菌种的最小抑菌浓度(MIC)值。
分析唑啉酮衍生物(6a-e,10a-d,30a-n,35a-c)对6种细菌的抑菌活性。结果表明,针对大肠杆菌(ATCC25922),粪肠球菌(ATCC29212),肠炎沙门杆菌(87),标准金黄色葡萄球菌(ATCC29213),耐甲氧西林金黄色葡萄球菌MRSA(R3708),绿脓杆菌(ATCC27853)这六种感染性细菌,喹唑啉酮类化合物(化合物30a,30d,35a)均有较显著的活性。
表3 化合物30a,30d,35a抗菌效力
注:aNT表示未测试。
试验例5
化合物30d与SeThrRS的复合物晶体的制备、X射线衍射数据收集与共晶结构解析。
(1)SeThrRS与SeThrRS-化合物30d复合物的晶体的制备
晶体采用坐滴气相扩散法生长。化合物浓度为4mM,SeThrRS蛋白浓度为20mg/mL。结晶条件为0.22M醋酸锂pH 7.0,20%(w/v)PEG3350,在25℃生长2天,即可获得可供X射线衍射的高分辨率晶体。
(2)SeThrRS与SeThrRS-化合物30d复合物的晶体X射线衍射数据采集与共晶结构解析
衍射数据采集在上海同步辐射光源(SSRF)BL19U1工作站与中山大学药学院室光源进行。每颗晶体收集180幅衍射画面,每幅图像曝光时间为0.5秒,旋转角度1°。SeThrRS与SeThrRS-化合物30d复合物的两颗晶体均采集到分辨率的完整衍射数据。
衍射数据采用HKL2000软件与CrysAlis Pro进行指标化、积分和合并。利用MOLREP程序,以大肠杆菌ThrRS的三维结构座标(PDB编号:4HWP)为模板,通过分子置换解析衍射相位。利用Coot程序,根据实空间的电子密度图形状,手动修正和完善蛋白质结构模型。同时,使用Refmac5程序,在倒易空间对结构模型进行自动优化。在实空间与倒易空间之间,交替进行多轮的结构修正,直至结构模型达到较高质量。关于数据采集和结构修正的主要统计参数见表4。晶体结构显示化合物30d作用于SeThrRS的tRNA和氨基酸位点。图3为SeThrRS三维结合模式图;图4为SeThrRS-化合物30d复合物三维结合模式图。
表4 SeThrRS与SeThrRS-化合物30d的晶体收集修正统计表
a括号内为最高分辨率壳层的数据。
b计算公式为:数据合并的线性R因子=∑h∑k|Ihk-<Ih>|/∑h∑k<Ih>,其中,Ihk是第h个衍射点第k次重复的衍射强度,<Ih>是第h个衍射点重复收集多次后的平均值。
c工作R因子=∑h||Fobs(h)|-|Fcal(h)||/∑h|Fobs(h)|,其中Fobs(h)和Fcal(h)分别表示h衍射点实验观察到的和依据结构模型计算得到的结构因子。
d自由R因子的计算方法与工作R因子一致,但自由R因子是由随机挑选出的5%的专用于监测的衍射点来计算的。
显然,本发明的上述实施例仅仅是为清楚地说明本发明所作的举例,而并非是对本发明的实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等,均应包含在本发明权利要求的保护范围之内。
Claims (10)
2.根据权利要求1所述的喹唑啉酮类化合物,其特征在于,R1为氢、氟、氯、溴、甲基、甲氧基或硝基。
3.根据权利要求1所述的喹唑啉酮类化合物,其特征在于,R2为氢、氟、氯、溴、甲基或甲氧基。
4.根据权利要求1所述的喹唑啉酮类化合物,其特征在于,R3为氢、甲基、乙基或异丙基。
5.根据权利要求1所述的喹唑啉酮类化合物,其特征在于,R4为氢或甲基。
6.根据权利要求1所述的喹唑啉酮类化合物,其特征在于,R5为C3~C6烷基、C4烯烃基或烷烃磺酰基;优选地,R5为-(CH2)3-、-(CH2)4-、-(CH2)5-、-(CH2)6-、(E)-异-2-丁烯基或丙烷磺酰基。
7.权利要求1~6任一项所述喹唑啉酮类化合物的制备方法,其特征在于,将含喹唑啉酮衍生基团的第一反应物与反应后取代基为X1基团的第二反应物、L-苏氨酸衍生物反应,得到所述喹唑啉酮类化合物。
8.权利要求1~6任一项所述喹唑啉酮类化合物在制备抗细菌感染药物中的应用。
9.根据权利要求8所述的应用,其特征在于,所述抗细菌感染药物的靶标为细菌型苏氨酰转运核糖核酸合成酶。
10.根据权利要求8或9所述的应用,其特征在于,所述抗细菌感染药物为抗大肠杆菌、肠炎沙门杆菌、绿脓杆菌、标准金黄色葡萄球菌、耐甲氧西林金黄色葡萄球菌MRSA或粪肠球菌药物。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113181195A (zh) * | 2021-04-07 | 2021-07-30 | 中山大学 | 葡萄糖多酚酸酯类衍生物在制备异亮氨酰tRNA合成酶抑制剂中的应用 |
CN116082314A (zh) * | 2023-02-27 | 2023-05-09 | 中山大学 | 一种喹唑啉类化合物及其制备方法和在制备脯氨酰tRNA合成酶抑制剂中的应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1123028A (zh) * | 1993-05-12 | 1996-05-22 | 纳幕尔杜邦公司 | 杀真菌的稠合双环嘧啶酮类化合物 |
CN107257789A (zh) * | 2015-02-13 | 2017-10-17 | 牛津药物设计有限公司 | 作为氨酰‑trna合成酶抑制剂的新型n‑酰基‑芳基磺酰胺衍生物 |
CN110041306A (zh) * | 2018-01-15 | 2019-07-23 | 中国医学科学院药物研究所 | 4-羟基-2-喹诺酮-氮-(4-喹唑啉酮)-3-甲酰胺类衍生物 |
-
2019
- 2019-11-15 CN CN201911122309.7A patent/CN110950863B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1123028A (zh) * | 1993-05-12 | 1996-05-22 | 纳幕尔杜邦公司 | 杀真菌的稠合双环嘧啶酮类化合物 |
CN107257789A (zh) * | 2015-02-13 | 2017-10-17 | 牛津药物设计有限公司 | 作为氨酰‑trna合成酶抑制剂的新型n‑酰基‑芳基磺酰胺衍生物 |
CN110041306A (zh) * | 2018-01-15 | 2019-07-23 | 中国医学科学院药物研究所 | 4-羟基-2-喹诺酮-氮-(4-喹唑啉酮)-3-甲酰胺类衍生物 |
Non-Patent Citations (1)
Title |
---|
KADALLIPURA PUTTASWAMY RAKESH 等: "Effect of Low Charge and High Hydrophobicity on Antimicrobial Activity of the Quinazolinone-Peptide Conjugates", 《RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113181195A (zh) * | 2021-04-07 | 2021-07-30 | 中山大学 | 葡萄糖多酚酸酯类衍生物在制备异亮氨酰tRNA合成酶抑制剂中的应用 |
CN116082314A (zh) * | 2023-02-27 | 2023-05-09 | 中山大学 | 一种喹唑啉类化合物及其制备方法和在制备脯氨酰tRNA合成酶抑制剂中的应用 |
CN116082314B (zh) * | 2023-02-27 | 2024-05-31 | 中山大学 | 一种喹唑啉类化合物及其制备方法和在制备脯氨酰tRNA合成酶抑制剂中的应用 |
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