CN108503658A - 一种近红外氯化氮杂氟硼烷染料及其制备方法和应用 - Google Patents
一种近红外氯化氮杂氟硼烷染料及其制备方法和应用 Download PDFInfo
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- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
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Abstract
本发明公开了一种近红外氯化氮杂氟硼烷染料。所述染料在制备过程中经由烯酮的合成‑加成反应‑成环反应‑配位取代反应等多步反应最终使得氯取代基本的氮杂氟硼烷骨架。所述染料在近红外波长区域有较强的吸收和发射,光热性质和化学稳定性都有提高,可用于光热成像、光声成像引导下的光动力治疗和光热治疗、生物标记和检测、时间门成像技术等领域。
Description
技术领域
本发明属于有机光电材料技术领域。具体涉及一种基于氮杂氟硼烷的具有近红外吸收的光热染料和其制备方法及其在荧光成像、光热成像、光声成像、光热治疗以及光动力治疗等领域中的应用。
背景技术
肿瘤一直是难以攻克的致死性疾病,目前对肿瘤的治疗方法还停留在传统的手术疗法、化疗或放射疗法;这些治疗方式往往会伴随着毒副作用的产生,治疗效果不佳。探索新的、毒副作用小的治疗方法,或者在已有治疗方法的基础上增加一些辅助手段,进而提高疗效、减轻毒副作用,是目前肿瘤治疗亟待解决的问题。
随着科学技术的不断发展,科研工作者逐渐开发出了多种新型治疗方案,其中光热光动力治疗作为一种非侵入性的肿瘤治疗手段,可以实时的进行肿瘤部位精确治疗,大大的提高了肿瘤治疗的效果。主要是将吲哚菁绿(ICG)、卟啉衍生物和二氢卟吩-e6(Ce6)等光敏剂呈递到肿瘤部位,在外部光源照射下,光敏剂能够吸收光能,并将氧气转化为有细胞毒性的活性氧。但是,光热光动力作为一个整体,其能量分配方式存在了相互竞争的关系,如何选择合适的光敏剂已经成为人们研究的重点。
近红外染料是一类性能良好的功能性染料,在近红外光区有很好的吸收,在无线电射频识别、肿瘤治疗、太阳能电池、印刷防伪等领域有广泛的应用。用于生物成像时,除了具有近红外吸收、发射波长、还具有良好的水溶性和较低的生物毒性,特异的组织或细胞靶向性以及良好的细胞穿透性等,从而达到更安全、高效、灵敏的荧光成像目的。
近红外染料包括菁染料类、罗丹明类、方酸类、卟啉类和BODIPY类,其中,氟硼烷类染料作为一种常用的染料具有好的光稳定性、高的量子产率、大的摩尔消光系数并且易修饰。这也使得其广泛应用于荧光标记、光学成像、光学肿瘤治疗等领域。与具有相似结构的氟硼烷相比,氮杂氟硼烷具有更长的吸收波长,在活体应用中独具优势,且其展现出不可忽略的辐射跃迁,这部分辐射跃迁会削弱单线态到三线态的转变使其不仅有良好的光热效应,还拥有了一定的光动力效果。但是如何实现其光热光动力效果的平衡依然是一个重要的研究方向。
氮杂氟硼烷染料一般用于生物标记、成像、光动力治疗等领域,目前为止,关于如何平衡其光热光动力效果以使其能量利用效率达到最好的报道还较少,因此有必要设计合成具有良好光热和光动力相互协同效果的近红外染料来解决现有技术中存在的不足。
发明内容
本发明的目的在于解决现有技术中的不足,设计并合成一种近红外氯化氮杂氟硼烷染料,基于光致电子转移的理论指导和氮杂氟硼烷类染料的优异性能,将氯原子连接到具有近红外吸收的氮氟硼烷染料上,从而使其具有良好的光声成像、光热成像及生物肿瘤的光热和光动力治疗效果。
本发明的技术方案为:本发明提供了一种近红外氯化氮杂氟硼烷染料,其特征在于,所述染料的结构通式如下:
其中,R1、R2选自下列中的任一个
其中,R3为具有1至16个碳原子的直链、支链或者环状烷基链。
所述近红外氯化氮杂氟硼烷染料的具体合成路线如下:
进一步地,所述的近红外氯化氮杂氟硼烷染料的具体合成步骤为:
1)烯酮的合成:将化合物1与2在10%-50%氢氧化钠和乙醇的混合溶液中反应过夜后,用稀盐酸调节反应液的酸碱度至pH值在5~7之间,过滤、冷乙醇洗涤,得到产物3;
2)加成反应:将所述产物3、硝基甲烷和二乙胺溶于无水甲醇中,加热回流16-28h,用稀盐酸调节反应液的酸碱度至pH值在5~7之间,萃取,减压蒸发溶剂,在二氯甲烷和***中进行重结晶,得到产物4;
3)成环反应:所述产物4在正丁醇溶液中与乙酸铵加热回流40-50h,冷却、过滤、冷乙醇洗涤,得到产物5;
4)配位取代反应:将所述产物5溶于二氯甲烷溶液中,向该溶液中加入N,N-二异丙基乙胺,在冰浴条件下进行反应,向其中滴加三氟化硼***,萃取,减压蒸发溶剂,得到产物6;将1-氯-1,2-苯碘酰-3-酮与所述产物6溶于二氯甲烷或乙腈溶液中,室温下搅拌进行反应,浓缩,石油醚和乙酸乙酯通过柱色谱法纯化,得到目标产物7。
进一步地,所述近红外氯化氮杂氟硼烷染料可应用在肿瘤的光动力和光热治疗中。
本发明的有益效果是:
1.本发明所述的近红外氯化氮氟硼烷染料的制备及分离提纯过程相对简单、原料来源丰富、产率较高,制备的染料有较高的化学稳定性;
2.本发明所述的近红外氯化氮氟硼烷染料的吸收和发射波长位于近红外光区,削弱了激发光源对生物组织的伤害,削弱背景荧光对检测信号的影响,具有较深的组织穿透深度;
3.本发明所述的近红外氯化氮氟硼烷染料可用于光热成像、光声成像引导下的肿瘤治疗,是很好的生物光热光动力治疗材料。
附图说明
图1是本发明实施例2中测定的化合物7的MALDI-TOF图;
图2是本发明实施例3中测定的化合物7的1H-NMR图;
图3是本发明实施例4中测定的化合物7的紫外-可见光谱图;
图4是本发明实施例5中测定的化合物7的发射光谱图;
图5是本发明实施例6中测定的不同浓度的化合物7的光热效果图;
图6是本发明实施例7中测定的不同浓度的化合物7的光动力效果图。
具体实施方式
以下实施例进一步说明本发明的内容,但不应理解为对本发明的限制。在不背离本发明实质的情况下,对本发明方法、步骤或条件所作的修改和替换,均属于本发明的范围。
实施例1:氯原子取代氮杂氟硼烷的合成,具体的合成路线如下:
化合物1的合成
取一洁净两口瓶,加入磁子、3.40g的对羟基苯乙酮(约25mmol)、24.14g的溴辛烷(约125mmol)、17.25g碳酸钾和20mL的无水N,N-二甲基甲酰胺溶液。磁力搅拌下,80℃反应24小时。反应结束后,水/二氯甲烷萃取多次,合并有机相。经层析柱分离,得到无色液体(产率99%)。
化合物2的合成
取一洁净两口瓶,加入磁子、3.05g的对羟基苯甲醛(约25mmol)、24.14g的溴辛烷(约125mmol)、17.25g碳酸钾20mL的无水N,N-二甲基甲酰胺溶液。磁力搅拌下,80℃反应24小时。反应结束后,水/二氯甲烷萃取多次,合并有机相。经层析柱分离,得到浅黄色液体(产率约99%)。
化合物3的合成
向溶有化合物1(3.73g,15mmol)和化合物2(3.15mg,15mmol)的乙醇(100mL)溶液中滴加质量分数为10%的氢氧化钠溶液(15mL)并在室温下搅拌24小时。在反应过程中,产物从反应混合物中沉淀出来。过滤反应混合物,所得固体用冷乙醇洗涤,得到黄色固体(6.34g,90.9%)。
1H NMR(400MHz,CDCl3)d(ppm)δ8.10–7.94(m,2H),7.77(d,J=15.6Hz,1H),7.64–7.51(m,2H),7.43(d,J=15.6Hz,1H),7.03–6.94(m,4H),4.04–3.97(m,4H),1.88–1.74(m,4H),1.49–1.25(m,20H),0.89(t,J=13.6Hz,6H).
化合物4的合成
将化合物3(6.04g,13mmol),硝基甲烷(3.97g,65mmol)和二乙胺(4.76g,65mmol)溶于无水甲醇(100mL)中并加热回流24小时。将该溶液冷却,用1M HCl酸化,然后用CH2Cl2/H2O萃取。合并的有机层经无水Na2SO4干燥。减压蒸发溶剂。所得粗产物从二氯甲烷和***中重结晶。得到化合物4,为白色固体(4.92g,71.9%)。
1H NMR(400MHz,CDCl3)d(ppm):δ7.88(d,J=8.0Hz,2H),7.17(d,J=8.0Hz,2H),6.90(d,J=8.3Hz,2H),6.83(d,J=7.6Hz,2H)4.81–4.60(m,2H),4.18–4.11(m,1H),4.00(t,J=12.8,2H),3.90(t,J=12.8,2H),3.41–3.28(m,2H),1.82–1.72(m,4H),1.50–1.28(m,20H),0.88(t,J=10.8Hz,6H).
化合物5的合成
将化合物4(6.00g,11.4mmol)和乙酸铵(26.3g,0.38mol)在正丁醇(150mL)中加热回流48小时。将溶液冷却并过滤沉淀物。将固体用冷乙醇洗涤并分离成黑色固体。该固体直接用于下一步。
化合物6的合成
将化合物5(3.85g,4mmol)溶于二氯甲烷(100mL)中,向该溶液中加入N,N-二异丙基乙胺(2.58g,20mmol),然后将混合物在冰浴条件下搅拌2h。将三氟化硼醚合物(2.84g,20mmol)滴加到混合物中。该溶液用水淬灭,然后用CH2Cl2/H2O提取。合并的有机层经无水Na2SO4干燥。减压蒸发溶剂。得到黑色固体(3.86g,95.6%)。
1H NMR(400MHz,CDCl3)d(ppm):δ8.05(d,J=2.0,4H),8.03(d,J=2.0,4H)6.98(d,J=8.8Hz,8H,),6.92(s,2H),4.03(q,J=19.2Hz,8H),1.86–1.78(m,8H),1.51–1.44(m,8H),1.37–1.28(m,32H),1.11(s,12H)0.90(t,J=13.6 12H,).
化合物7的合成
根据已有合成方案合成1-氯-1,2-苯并碘代-3-酮,将HABM(202.0mg,0.2mmol)和1-氯-1,2-苯并碘代-3-酮(62.2mg,0.22mmol))溶于乙腈(30mL)中,将混合物在室温下搅拌12小时。然后用CH2Cl2萃取混合物,用水洗涤,用石油醚和乙酸乙酯通过柱色谱法纯化,得到黑色固体(164.9mg,74.6%)。
1H NMR(400MHz,CDCl3)d(ppm):δ7.97–7.90(m,4H),7.81–7.75(m,4H),7.01–6.93(m,8H),4.05–3.99(m,8H),1.87–1.76(m,8H),1.53–1.42(m,8H),1.40–1.25(m,32H),0.92–0.88(m,12H).
实施例2:化合物7的分子量的测试
取少量化合物6,与基质混匀,然后点样,用MALDI–TOF进行测定,结果如图1,初步证明了化合物7分子的正确性。
[m/e](M,MALDI-TOF)理论值:1077.59,实验值:1078.12。
实施例3:化合物7的核磁测试
取0.5mg化合物7溶于0.5mL氘代氯仿中,经核磁测试,结果如图2,进一步证明了化合物7分子的正确性。
1H NMR(400MHz,CDCl3)d(ppm):δ7.97–7.90(m,4H),7.81–7.75(m,4H),7.01–6.93(m,8H),4.05–3.99(m,8H),1.87–1.76(m,8H),1.53–1.42(m,8H),1.40–1.25(m,32H),0.92–0.88(m,12H).
13C NMR(100MHz,CDCl3)d(ppm):δ161.32,160.39,155.60,143.21,138.92,132.35,122.83,120.96,114.16,68.11,31.85,29.55,29.10,26.08,22.69.
19F NMR(376.5MHz,CDCl3):d(ppm):δ-131.51(q,2F).MALDI-TOF-MS(m/z):calcdfor C64H84BCl2F2N3O4,1079.10;found,1077.47.
实施例4:化合物7的紫外-可见光谱测试:
配置实施例1中的化合物7的稀溶液(10-5M,二氯甲烷为溶剂),移取2mL化合物7溶液于比色皿中进行紫外-可见光谱测试,结果如图3,从图3中可以看出随着引入供电子基团后,实现了更大的近红外吸收。
实施例5:化合物7的发射光谱测试:
配置实施例1中的化合物7的稀溶液(10-5M,二氯甲烷为溶剂),移取2mL化合物7的稀溶液于比色皿中进行发射光谱测试。结果如图4,从图4中可以看出化合物7在甲苯溶液中具有很好的近红外发射,最大发射波长是785nm。
实施例6:化合物7的光热效果测试:
将实施例1中的化合物7用二氯甲烷溶解,二氯甲烷溶液将其稀释成不同浓度梯度的溶液(10、20、30μM),用波长为660nm激光器照射5min(0.4W/cm2),用光热成像仪记录不同时间、不同浓度下,溶液温度的变化。结果如图5,从图5中可以看出,随着浓度增加,化合物7的溶液温差最高可达23℃,说明该化合物具有很好的光热效果。
实施例7:化合物7的光动力效果测试:
将实施例1中的化合物7用二氯甲烷溶解,DBPF的二氯甲烷溶液作为单线态氧指示剂,将其稀释成不同浓度梯度的溶液(0、5、10、15、25μM),用波长为690nm的激光器照射7min(0.4W/cm2),用光热成像仪记录不同时间、不同浓度下,溶液温度的变化。由图6表明,随着照射时间的改变,DPBF的吸收强度也随之变化,表明化合物7能够有效的产生单线态氧,可用于光动力治疗。
以上显示和描述了本发明的基本原理、主要特征及优点。但是以上所述仅为本发明的具体实施例,本发明的技术特征并不局限于此,任何本领域的技术人员在不脱离本发明的技术方案下得出的其他实施方式均应涵盖在本发明的专利范围之中。
Claims (5)
1.一种近红外氯化氮杂氟硼烷染料,其特征在于,所述染料的结构通式如下:
其中,R1、R2选自下列中的任一个
其中,R3为具有1至16个碳原子的直链、支链或者环状烷基链。
2.如权利要求1所述的一种近红外氯化氮杂氟硼烷染料的制备方法,其特征在于,具体合成路线如下:
3.如权利要求2所述的一种近红外氯化氮杂氟硼烷染料的制备方法,其特征在于,具体合成步骤为:
1)烯酮的合成:将化合物1与2在10%-50%氢氧化钠和乙醇的混合溶液中反应过夜后,用稀盐酸调节反应液的酸碱度至pH值在5~7之间,过滤、冷乙醇洗涤,得到产物3;
2)加成反应:将所述产物3、硝基甲烷和二乙胺溶于无水甲醇中,加热回流16-28h,用稀盐酸调节反应液的酸碱度至pH值在5~7之间,萃取,减压蒸发溶剂,在二氯甲烷和***中进行重结晶,得到产物4;
3)成环反应:所述产物4在正丁醇溶液中与乙酸铵加热回流40-50h,冷却、过滤、冷乙醇洗涤,得到产物5;
4)配位取代反应:将所述产物5溶于二氯甲烷溶液中,向该溶液中加入N,N-二异丙基乙胺,在冰浴条件下进行反应,向其中滴加三氟化硼***,萃取,减压蒸发溶剂,得到产物6;将1-氯-1,2-苯碘酰-3-酮与所述产物6溶于二氯甲烷或乙腈溶液中,室温下搅拌进行反应,浓缩,石油醚和乙酸乙酯通过柱色谱法纯化,得到目标产物7。
4.如权利要求1-3中任一项所述的近红外氯化氮杂氟硼烷染料在肿瘤的光动力治疗中的应用。
5.如权利要求1-3中任一项所述的近红外氯化氮杂氟硼烷染料在肿瘤的光热治疗中的应用。
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CN113912762B (zh) * | 2021-09-24 | 2023-02-10 | 复旦大学 | 水溶性的近红外二区大分子荧光探针及其制备方法和应用 |
CN113980039A (zh) * | 2021-11-19 | 2022-01-28 | 杭州师范大学 | 一种光热剂及其制备方法和应用 |
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