CN112707921A - 一种酶抑制剂的荧光标记的合成 - Google Patents

一种酶抑制剂的荧光标记的合成 Download PDF

Info

Publication number
CN112707921A
CN112707921A CN201911020611.1A CN201911020611A CN112707921A CN 112707921 A CN112707921 A CN 112707921A CN 201911020611 A CN201911020611 A CN 201911020611A CN 112707921 A CN112707921 A CN 112707921A
Authority
CN
China
Prior art keywords
cox
rhodamine
solution
mercaptoguanine
reaction solution
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201911020611.1A
Other languages
English (en)
Inventor
武松宇
任深圳
叶亚熙
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute Of Artificial Intelligence Biomedical Technology Nanjing University
Nanjing Carbon Silicon Artificial Intelligence Biomedical Technology Research Institute Co Ltd
Original Assignee
Institute Of Artificial Intelligence Biomedical Technology Nanjing University
Nanjing Carbon Silicon Artificial Intelligence Biomedical Technology Research Institute Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute Of Artificial Intelligence Biomedical Technology Nanjing University, Nanjing Carbon Silicon Artificial Intelligence Biomedical Technology Research Institute Co Ltd filed Critical Institute Of Artificial Intelligence Biomedical Technology Nanjing University
Priority to CN201911020611.1A priority Critical patent/CN112707921A/zh
Publication of CN112707921A publication Critical patent/CN112707921A/zh
Pending legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D519/00Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/06Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10Non-macromolecular compounds
    • C09K2211/1018Heterocyclic compounds
    • C09K2211/1025Heterocyclic compounds characterised by ligands
    • C09K2211/1029Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
    • C09K2211/1033Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom with oxygen
    • CCHEMISTRY; METALLURGY
    • C09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10Non-macromolecular compounds
    • C09K2211/1018Heterocyclic compounds
    • C09K2211/1025Heterocyclic compounds characterised by ligands
    • C09K2211/1074Heterocyclic compounds characterised by ligands containing more than three nitrogen atoms as heteroatoms

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Optics & Photonics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Engineering & Computer Science (AREA)
  • Materials Engineering (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

本发明本发明将非甾体抗炎药吲哚美辛转与罗丹明B反应形成缀合物,该缀合物能够选择性和有效地抑制两种纯化蛋白中的COX‑2和完整的细胞。这些不会显着抑制COX‑1的结合物可在体内炎症性病变和表达COX‑2的肿瘤中对COX‑2进行光学成像。该研究用于优化来自多种NSAID或COX‑2选择性抑制剂(COXIB)的荧光缀合物。将缀合物评估为靶向COX‑2的成像剂,并在体外和体内验证活性分子作为靶向COX‑2的细胞和肿瘤制剂。这为我们提供了一种荧光标记的光学成像剂,该荧光剂经验证可用于靶向COX‑2的炎症和癌症体内成像。本发明以吲哚美辛为原料,经与己二胺缩合,所得产物再与罗丹明B进行缩合。本发明提供了一种操作简便、原料易得,杂质含量少、生产成本低的在COX‑2抑制剂吲哚美辛上引入荧光基团的合成方法,在保证其基本骨架结构的基础上引入荧光基团,该化合物可自发荧光,为其在细胞生物学中的示踪应用。

Description

一种酶抑制剂的荧光标记的合成
技术领域
本发明属于药物化学领域
背景技术
叶酸是所有生物体生长的必需维生素。还原形式四氢叶酸是合成嘌呤,氨基酸,S-腺苷甲硫氨酸,胸苷单磷酸和甲酰基甲硫氨酸所需的碳转移反应的关键辅助因子。哺乳动物和高等真核生物依赖于饮食中的叶酸,而植物和大多数微生物是从头合成的。靶向叶酸合成途径的二氢蝶呤合酶(DHPS)的磺胺药物在数十年后仍在临床中使用,即使在今天,磺胺类药物仍与细菌二氢叶酸还原酶(DHFR)抑制剂共同使用,作为一种协同的广谱抗菌药物,可预防或治疗多种疾病和感染,包括疟疾,弓形虫脑炎,卡氏肺孢子虫肺炎,肺结核和金黄色葡萄球菌感染。
金黄色葡萄球菌的耐甲氧西林菌株(MRSA)(“超级细菌”)的出现突出了抗生素耐药性问题的严重性。最初仅限于医院环境,最近又传播到社区,并开始影响那些没有任何危险因素的人。寻找有效的未来治疗方法已引起人们对新型抗菌药物的替代酶标靶以及新型免疫策略的兴趣。有文献报道基于6-羟甲基-7,8-二氢蝶呤焦磷酸激酶(HPPK)抑制剂的开发,该酶是叶酸合成途径中位于DHPS之前的酶,负责催化焦磷酸酰基从结合镁的ATP辅助因子转移至6-羟甲基-7,8-二氢蝶呤(HMDP)。HPPK不是任何现有抗生素的已知靶标,没有相关的人类同系物,并且具有可开发可对当前和未来的SMX/TMP耐药病原体有效的抗菌药物的位点,目前发现8-巯基鸟嘌呤(8MG)是金黄色葡萄球菌HPPK(SaHPPK)的抑制剂。
如上所述,6-羟甲基-7,8-二氢蝶呤焦磷酸激酶(HPPK)抑制剂有着如此巨大的科研和生物医学潜力,但其作用机理并没有完全清晰,本发明在8-巯基鸟嘌呤结构的基础上引入罗丹明B荧光基团,使8-巯基鸟嘌呤可自发荧光,以便被示踪。
发明内容
本发明要解决的技术问题在于提供一种操作简便、原料易得,杂质含量少、生产成本低的8-巯基鸟嘌呤罗丹明合物的合成方法。
本发明提供的8-巯基鸟嘌呤罗丹明合物的技术方案如下:
罗丹明B酰肼的制备方法:
将罗丹明B(2g,4.2mmol)溶解于15mL乙醇的溶液中,随后加入过量的水合肼(2.5mL),然后将反应溶液回流直至粉红色消失。之后,将冷却的反应溶液倒入蒸馏水中,并用乙酸乙酯(3×30mL)萃取。合并的萃取物用无水硫酸钠干燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱法进一步纯化,用石油醚/CH2Cl2=1∶1洗脱,得到罗丹明B酰肼(1.0g,2.2mmol),为白色固体。产率:52.4%。反应式如下:
Figure BSA0000193118250000021
在0℃下向溶解于15mL无水CHCl3中的罗丹明B酰肼(815mg,1.8mmol)的溶液中添加三乙胺(332μL,2.4mmol),并将反应溶液搅拌几分钟。然后,滴加2-氯乙酰氯(180μL,2.4mmol)于5mL无水CHCl3中的溶液。之后,将混合物加热至室温并搅拌4小时。然后将反应溶液倒入蒸馏水中,并用CH2Cl2(3×30mL)萃取。合并的有机层依次用饱和NaCl水溶液(50mL)洗涤,并用无水Na2SO4干燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱进一步纯化,用石油醚/乙酸乙酯=3∶1洗脱,得到Rh-2(860mg,1.6mmol),为白色固体。产率:88.9%。反应式如下:
Figure BSA0000193118250000022
2.将罗丹明荧光基团引入6-羟甲基-7,8-二氢蝶呤焦磷酸激酶(HPPK)抑制剂8-巯基鸟嘌呤中,其具体合成方法为:
将Rh-2(530mg,1mmol)溶解于15m四氢呋喃中,随后加入过量的三乙胺(3mmol),再加入8-巯基鸟嘌呤,随后加热回流,反应过夜。之后,将冷却的反应溶液倒入蒸馏水中,并用乙酸乙酯(3×30mL)萃取。合并的萃取物用无水硫酸钠干燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱法进一步纯化,用CH2Cl2/MeOH=20∶1洗脱,得到8-巯基鸟嘌呤罗丹明合物(366mg,0-54mmol),为白色固体。产率:54%。反应式如下:
Figure BSA0000193118250000031
3.上述的8-巯基鸟嘌呤罗丹明合物,其特征在于,该化合物可以自发荧光,应用于细胞生物学中的示踪。
具体实施方式
下面结合实施例对本发明作进一步说明,但本发明的保护范围并不限于此
实施例1
罗丹明B酰肼的合成:将罗丹明B(2g,4.2mmol)溶解于15mL乙醇的溶液中,随后加入过量的水合肼(2.5mL),然后将反应溶液回流直至粉红色消失。之后,将冷却的反应溶液倒入蒸馏水中,并用乙酸乙酯(3×30mL)萃取。合并的萃取物用无水硫酸钠干燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱法进一步纯化,用石油醚/CH2Cl2=1∶1洗脱,得到罗丹明B酰肼(1.0g,2.2mmol),为白色固体。1H NMR(400MHz,CDCl3)δ7.94(d,J=8.0Hz,1H),7.51-7.38(m,2H),7.11(d,J=7.4Hz,1H),6.46(d,J=8.8Hz,2H),6.42(s,2H),6.29(d,J=8.8Hz,2H),3.61(s,2H),3.34(q,J=7.0Hz,8H),1.17(t,J=7.0Hz,12H)。
Rh-2的合成:在0℃下向溶解于15mL无水CHCl3中的罗丹明B酰肼(815mg,1.8mmol)的溶液中添加三乙胺(332μL,2.4mmol),并将反应溶液搅拌几分钟。然后,滴加2-氯乙酰氯(180μL,2.4mmol)于5mL无水CHCl3中的溶液。之后,将混合物加热至室温并搅拌4小时。然后将反应溶液倒入蒸馏水中,并用CH2Cl2(3×30mL)萃取。合并的有机层依次用饱和NaCl水溶液(50mL)洗涤,并用无水Na2SO4干燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱进一步纯化,用石油醚/乙酸乙酯=3∶1洗脱,得到Rh-2(860mg,1.6mmol),为白色固体。产率:88.9%。1H NMR(400MHz,DMSO)δ9.92(s,1H),7.83(d,J=7.6Hz,1H),7.62-7.48(m,2H),7.02(d,J=6.7Hz,1H),6.55-6.45(m,2H),6.34(dd,J=7.2,2.3Hz,4H),4.00(s,2H),3.32(q,J=7.0Hz,8H),1.08(t,J=7.0Hz,12H)。
8-巯基鸟嘌呤罗丹明合物的合成:将Rh-2(530mg,1mmol)溶解于15m四氢呋喃中,随后加入过量的三乙胺(3mmol),再加入8-巯基鸟嘌呤,随后加热回流,反应过夜。之后,将冷却的反应溶液倒入蒸馏水中,并用乙酸乙酯(3×30mL)萃取。合并的萃取物用无水硫酸钠干燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱法进一步纯化,用CH2Cl2/MeOH=20∶1洗脱,得到8-巯基鸟嘌呤罗丹明合物(366mg,0-54mmol),为白色固体。产率:54%。1H NMR(600MHz,DMSO-d6)δ10.50(s,1H),8.69(s,1H),7.82-7.76(m,1H),7.52-7.45(m,2H),7.01-6.95(m,1H),6.36(s,5H),6.04(s,2H),3.49(s,2H),3.34-3.24(m,7H),3.08(t,J=7.3Hz,2H),2.78(q,J=6.7Hz,2H),1.06(t,J=7.0Hz,12H)。

Claims (3)

1.罗丹明B酰肼的制备方法:
将罗丹明B(2g,4.2mmol)溶解于15mL乙醇的溶液中,随后加入过量的水合肼(2.5mL),然后将反应溶液回流直至粉红色消失。之后,将冷却的反应溶液倒入蒸馏水中,并用乙酸乙酯(3×30mL)萃取。合并的萃取物用无水硫酸钠于燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱法进一步纯化,用石油醚/CH2Cl2=1∶1洗脱,得到罗丹明B酰肼(1.0g,2.2mmol),为白色固体。产率:52.4%。反应式如下:
Figure FSA0000193118240000011
在0℃下向溶解于15mL无水CHCl3中的罗丹明B酰肼(815mg,1.8mmol)的溶液中添加三乙胺(332μL,2.4mmol),并将反应溶液搅拌几分钟。然后,滴加2-氯乙酰氯(180μL,2.4mmol)于5mL无水CHCl3中的溶液。之后,将混合物加热至室温并搅拌4小时。然后将反应溶液倒入蒸馏水中,并用CH2Cl2(3×30mL)萃取。合并的有机层依次用饱和NaCl水溶液(50mL)洗涤,并用无水Na2SO4干燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱进一步纯化,用石油醚/乙酸乙酯=3∶1洗脱,得到Rh-2(860mg,1.6mmol),为白色固体。产率:88.9%。反应式如下:
Figure FSA0000193118240000012
2.根据权利要求1所述的Rh-2的合成方法,其特征在于,将罗丹明荧光基团引入6-羟甲基-7,8-二氢蝶呤焦磷酸激酶(HPPK)抑制剂8-巯基鸟嘌呤中,其具体合成方法为:
将Rh-2(530mg,1mmol)溶解于15m四氢呋喃中,随后加入过量的三乙胺(3mmol),再加入8-巯基鸟嘌呤,随后加热回流,反应过夜。之后,将冷却的反应溶液倒入蒸馏水中,并用乙酸乙酯(3×30mL)萃取。合并的萃取物用无水硫酸钠干燥。减压除去溶剂,并将残余物通过硅胶上的柱色谱法进一步纯化,用CH2Cl2/MeOH=20∶1洗脱,得到8-巯基鸟嘌呤罗丹明合物(366mg,0-54mmol),为白色固体。产率:54%。反应式如下:
Figure FSA0000193118240000021
3.根据权利要求2所述的8-巯基鸟嘌呤罗丹明合物的合成方法,其特征在于,该化合物可以自发荧光,应用于细胞生物学中的示踪。
CN201911020611.1A 2019-10-25 2019-10-25 一种酶抑制剂的荧光标记的合成 Pending CN112707921A (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201911020611.1A CN112707921A (zh) 2019-10-25 2019-10-25 一种酶抑制剂的荧光标记的合成

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201911020611.1A CN112707921A (zh) 2019-10-25 2019-10-25 一种酶抑制剂的荧光标记的合成

Publications (1)

Publication Number Publication Date
CN112707921A true CN112707921A (zh) 2021-04-27

Family

ID=75540440

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201911020611.1A Pending CN112707921A (zh) 2019-10-25 2019-10-25 一种酶抑制剂的荧光标记的合成

Country Status (1)

Country Link
CN (1) CN112707921A (zh)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110272437A (zh) * 2018-03-16 2019-09-24 中国科学院大连化学物理研究所 可见光光控的snap蛋白标签类耐酸荧光分子开关及其合成

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110272437A (zh) * 2018-03-16 2019-09-24 中国科学院大连化学物理研究所 可见光光控的snap蛋白标签类耐酸荧光分子开关及其合成

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ZI-YINGWU等: "Two novel rhodamine-based fluorescent probes for the rapid and sensitive detection of Fe3+: Experimental and DFT calculations", 《SPECTROCHIMICA ACTA PART A: MOLECULAR AND BIOMOLECULAR SPECTROSCOPY》 *

Similar Documents

Publication Publication Date Title
Wang et al. A novel DCM-NBD conjugate fluorescent probe for discrimination of Cys/Hcy from GSH and its bioimaging applications in living cells and animals
Wang et al. A general approach to spirolactonized Si-rhodamines
EP3564239B1 (en) Aryl hydrocarbon receptor modulator
EP2736916B1 (en) Minor groove binder phosphoramidites and methods of use
JP2016540801A (ja) フルオロフェニルピラゾール化合物
Tietz et al. Pyrimidine-based fluorescent COX-2 inhibitors: synthesis and biological evaluation
CN112707921A (zh) 一种酶抑制剂的荧光标记的合成
EP3542796B1 (en) Compound having anti-cancer effect, and preparation method therefor and use thereof
Kreituss et al. Discovery of aziridine-triazole conjugates as selective MMP-2 inhibitors
Kore et al. Concise synthesis of 5-methyl-, 5-formyl, and 5-carboxy analogues of 2′-deoxycytidine-5′-triphosphate
Zhou et al. Synthesis, biological evaluation and cellular localization study of fluorescent derivatives of Jiyuan Oridonin A
Diwakar et al. Synthesis, X-ray characterization and biological evaluation of some new 2-(4-methy-2-oxo-2 H-chromen-7yloxy) acetamide derivatives
CN112920167B (zh) 靶向fgfr和hdac的双靶点抑制剂及其制备方法和应用、药物组合物及药剂
CN111362873B (zh) 一种加替沙星代谢物的合成方法
CN111606888B (zh) 吡咯类衍生物及其制备方法与应用
EP3543228B1 (en) Compound having anticancer activity, and preparation method and application thereof
CN111440199A (zh) 大环类谷氨酰胺酶gls1抑制剂或其可药用的盐、其制备方法及用途
CZ309849B6 (cs) Použití derivátů akridinu jako sloučenin interkalujících se do DNA
Ghosh et al. Synthesis and evaluation of an imidazole derivative–fluorescein conjugate
WO2023284249A1 (zh) 咪唑吡嗪酮类荧光素及其制备方法
CN113999210B (zh) 一组2-苯氨基-4-三氮唑基嘧啶类衍生物及其应用
Zou et al. Photocaged probes for spatiotemporal imaging
ES2207296T3 (es) Sal sodica de 3-(4-cinamil-1-piperazinil)imino-metil rifamicina sv y procedimiento de preparacion.
CN110698533B (zh) 一种熊果酸吲哚醌基类衍生物及其制备方法和应用
US8404660B2 (en) Method of obtaining of 4-N-furfurylcytosine and/or its derivatives, an anti-aging composition and use of 4-N-furfurylcytosine and/or its derivatives in the manufacture of anti-aging composition

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20210427