生物荧光分析试剂Cy系列的应用及合成

经过多步合成,得到了生物荧光分析试剂菁染料Cy3,探讨了合成工艺,并对其应用进行了总结.     

基于Cy7类菁染料分子探针的设计策略

菁染料是一类经典的荧光染料母核, 具有摩尔消光系数大、 吸收波长可调、 溶解性良好及生物兼容性好等优点, 被广泛用于蛋白标记、 痕量金属离子检测、 生物活性物质检测、 细胞和活体成像及肿瘤靶向治疗等领域. 近年来, 生物医学领域对活体结构及功能成像深度提出更高的需求, 基于优异的长波长染料母核开发近红外荧光分子探针逐渐成为领域的研究重点. 吲哚七甲川菁染料(Cy7)是一类最具代表性的菁染料, 本文重点综合评述了自1992年以来基于Cy7结构开发的分子探针, 并介绍了该类荧光探针的设计策略. 最后, 讨论了该领域研究面临的挑战, 并对未来的发展方向进行了总结和展望.     

Cy5-Fe2+-H2O2光度法测定天然抗氧化剂的抗氧化活性

在缓冲溶液中,Fenton体系产生的羟基自由基能迅速与五甲川菁染料(Cy5)作用使其褪色,在647 nm处测定体系吸光度的变化,可间接测定羟基自由基的生成量。抗氧化剂的加入可部分消除溶液中的羟基自由基,从而使溶液的吸光度下降程度减弱。据此建立了一种测定抗氧化剂对羟基自由基清除率的新方法。利用所建立的方法测试了抗氧化剂2,6-二叔丁基对甲基苯酚以及几种天然抗氧化剂的抗氧化活性,得到了满意的结果。该方法操作简便、稳定性好,可以作为一种简便的筛选抗氧化剂的方法。     

Preparation and reaction of platinum(II) complexes of N,N-bis(dicyclohexylphosphinomethyl)aminomethane. Crystal structures of (Cy2PCH2)2NMe (Cy = cyclohexyl) and [PtX2{(Cy2PCH2)2NMe}], (X = Cl and I)

Treatment of [Cy₂P(CH₂OH)₂]Cl with MeNH₂ in the presence of Et₃N affords a high yield of the phosphine (Cy₂PCH₂)₂NMe (1) (dcpam) which has been characterised by a single crystal X-ray structure. Treatment of [PtX₂(COD)], (COD=cyclo-octa-1,5-diene, X= Cl or I) with (1) affords the platinum complexes [PtX₂{(Cy₂PCH₂)₂NMe}] (2). The chloride complex, (2a), reacts with t-BuNC to afford [PtCl(t-BuNC)-{(Cy₂PCH₂)₂NMe}]Cl (3) and treatment of (2a) with 2-mercapto-1-methylimidazole affords [Pt{SCN(Me)CHCH=N(Me)}{Cy₂PCH₂)₂NMe}]Cl (5). The reaction of (2a) with 2-acetamidoacrylic acid in the presence of silver(I) oxide affords the carbon bonded isomer (8a) only whereas a similar reaction using [PtCl₂{Ph₂P-(CH₂)₃PPh₂}] affords a mixture of the azaallyl complex (7) and the carbon bonded isomer (8b) which can be separated by fractional crystallisation. The crystal structures of PtX₂{(Cy₂PCH₂)₂NMe}] are also reported.     

中位醛基取代Cy5的光谱性能及作为粘度荧光探针的研究

中位醛基取代的五甲川菁染料(Cy5)与传统Cy5相比其最大吸收和发射波长稍微蓝移, 斯托克斯位移增加, 在水中能达到48 nm|具有低的荧光量子产率, 而且随溶剂极性变化较小. 实验发现这种染料对环境的粘度非常敏感, 随着染料分子所处环境的粘度逐渐增加, 其荧光强度明显增加, 而且荧光强度与溶液粘度的关系满足F?rster-Hoffmann关系式. 该染料具有活细胞膜的穿透性, 能跨膜进入活细胞内, 使得该染料在检测活细胞内微环境粘度变化方面具有潜在的应用价值.     

吲哚类菁染料Cy3嵌入的核壳荧光纳米颗粒的制备及其性质研究

以蛋白质或多肽修饰的吲哚类菁染料Cy3为内核, 采用实验条件简单的油包水反相微乳液方法成核, 通过正硅酸乙酯水解形成的网状二氧化硅包壳的方法制备吲哚类菁染料Cy3嵌入的核壳荧光纳米颗粒. 考察了以不同等电点的蛋白质和多肽修饰的Cy3为内核材料对吲哚类菁染料Cy3嵌入的核壳荧光纳米颗粒制备的影响. 结果表明, 分别采用人免疫球蛋白(IgG)或多聚赖氨酸修饰的Cy3为内核材料, 都能制备荧光强度高、荧光稳定性强和染料泄漏极少的Cy3嵌入的核壳荧光纳米颗粒. 进一步对Cy3嵌入的核壳荧光纳米颗粒进行了表征, 并将基于这一新型的荧光纳米颗粒建立起来的生物标记方法初步应用于流感病毒DNA的检测, 其检测线性范围为3.18×10-10～1.27×10-9 mol/L, 检测下限为3.51×10-10 mol/L, 相关系数r为0.986 5.     

CD20靶向Cy7-Rituximab分子探针的制备及在小鼠活体荧光成像中的应用

以B淋巴细胞表面CD20抗原靶向的单克隆抗体Rituximab为载体, 通过共价键偶联荧光基团菁染料Cy7, 获得了新型荧光分子探针Cy7-Rituximab. 利用全光谱紫外-可见分光光度仪、SDS-聚丙烯酰胺凝胶电泳和基质辅助激光解析电离飞行时间质谱等对该探针结构进行表征, 并通过激光共聚焦显微镜观察了其在弥漫大B细胞淋巴瘤(DLBCL)细胞中的摄取情况. 选用BALB/C裸鼠为模型, 尾静脉注射 Cy7-Rituximab, 通过活体荧光成像系统观察了其在小鼠体内的分布情况. 研究结果表明, 修饰后的Cy7-Rituximab保持了原有抗体的免疫活性. 活体荧光成像结果表明, 在CD20高表达的脾脏部位监测到该分子探针的特异性浓集.     

三烃基（Ph,Cy,Bu）锡芳硫（亚砜,砜）乙酸酯的合成及表征

合成了２２种三烃基（Ｐｈ，Ｃｙ，Ｂｕ）锡芳硫（亚砜，砚）乙酸酯，用元素分析、ＩＲ、ＨＮＭＲ、ＭＳ、ＸＰＳ等手段对其结构进行了表征，结果表明，三环己基锡芳硫（亚砜、砜）乙酸酯具有四配位锡原子的单体结构，三苯基（三丁基）锡芳硫（亚砜、砜）乙酸醌具有五配位原子的聚合结构，通过估算它们的正辛醇－水分配系数，预测其在环境中的吸附及生物富集能力。     

Studies on the reactivity of the clusters [Ru3(CO)8(μ-H)2(μ-PR2)2] (R=But,Cy) towards phosphine ligands

The reaction behavior of the 48e-clusters [Ru₃(CO)₈(μ-H)₂(μ-PR₂)₂] (R=Bu^t, 1a; R=Cy, 1b) towards phosphine ligands has been studied. Whereas 1a reacts spontaneously with many phosphines at room temperature, a lack of reactivity for 1b under similar conditions is observed. Thus 1a reacts with dppm (Ph₂PCH₂PPh₂) to the known 46e-cluster [Ru₃(μ-CO)(CO)₄(μ₃-H)(μ-H)(μ-PBu^t₂)₂(μ-dppm)] (2a), and the reaction of 1a with dppe (Ph₂PC₂H₄PPh₂) yields analogously [Ru₃(μ-CO)(CO)₄(μ₃-H)(μ-H)(μ-PBu^t₂)₂(μ-dppe)] (3). Reactions of 1a with dmpm (Me₂PCH₂PMe₂), dmpe (Me₂PC₂H₄PMe₂) and PBuⁿ₃, respectively, gave in each case a mixture of products which could not be characterized. Contrary to the reaction behavior at room temperature, 1b reacts with phosphines in THF under reflux yielding the novel complexes [Ru₃(CO)₆(μ-H)₂(μ-PCy₂)₂L₂] (L=Cy₂PH, 4a; L=Bu^t₂PH, 4b; L=Ph₂PH, 4c; L=P(OEt)₃, 4d). 4a is also obtained directly by the reaction of [Ru₃(CO)₁₂] with an excess of Cy₂PH. The molecular structure of 4a has been determined by a single-crystal X-ray analysis. Moreover, the thermolysis of 1a in octane affords [Ru₃(CO)₈(μ-H)₂(μ₃-PBu^t)(Bu^t₂PH)] (6) as the main product, and the thermolysis of [Ru₃(CO)₉(Bu^t₂PH)(μ-dppm)] (7) yields 2a to a considerable extent. Treatment of 1a with carbon tetrachloride leads to [Ru₃(CO)₇(μ-H)(μ-PBu^t₂)₂(μ-Cl)] (8) as the main product.     

Solid-State Confinement Effects in Selective exo-H/D Exchange in the Rhodium σ-Norbornane Complex [(Cy2PCH2CH2PCy2)Rh(η2:η2-C7H12)][BArF4]

Density functional theory calculations modelling selective exo-H/D exchange observed in the Rh σ-alkane complex [(Cy₂PCH₂CH₂PCy₂)Rh(η²:η²-endo-NBA)][BAr^F₄], [1-NBA][BAr^F ₄ ] , are reported, where Ar^F=3,5-C₆H₃(CF₃)₂ and NBA=norbornane, C₇H₁₂. Two models were considered 1) an isolated molecular cation, [1-NBA]⁺ and 2) a full model in which [1-NBA][BAr^F ₄ ] is treated in the solid state through periodic DFT. After an initial endo-exo rearrangement, both models predict H/D exchange to proceed through D₂ addition and oxidative cleavage followed by a rate-limiting C−H activation of the norbornane through a σ-CAM step to form a [1-Rh(D)(η²-HD)(norbornyl)]⁺ intermediate. HD rotation followed by a σ-CAM C−D bond formation, HD reductive coupling and HD loss then complete the H/D exchange process. exo-H/D exchange is facilitated by a supporting agostic interaction and is consistently more accessible kinetically than the potentially competing endo-H/D exchange (isolated cation: ΔG^≠_exo=+15.9 kcal/mol, ΔG^≠_endo=+18.4 kcal/mol; solid state: ΔG^≠_exo=+22.1 kcal/mol, ΔG^≠_endo=+25.1 kcal/mol). The solid-state environment has a significant impact on the computed energetics, with barriers increasing by ca. 7 kcal/mol, while only the solid-state model correctly predicts the endo-bound NBA complex to be the resting state of the system. These outcomes reflect solid-state confinement effects within the pocket occupied by the [1-NBA]⁺ cation and defined by the pseudo-octahedral array of neighbouring [BAr^F₄]⁻ anions. The asymmetry of the solid-state environment also requires a second H/D exchange pathway to be defined to account for reaction at all four exo-C−H bonds. These entail slightly higher barriers (ΔG^≠_exo= +24.8 kcal/mol, ΔG^≠_endo=+27.5 kcal/mol) but retain a distinct preference for exo- over endo-H/D exchange.     

Complexes with Dative Bonds between d‐ and s‐Block Metals: Synthesis and Structure of [(Cy3P)2Pt?Be(Cl)X] (X=Cl,Me)

A platinum–beryllium adduct (see structure) was prepared by the reaction of [Pt(PCy₃)₂] and BeCl₂. Treatment with methyllithium resulted in ligand substitution at the beryllium center. Both complexes were structurally characterized and display unprecedented two‐center two‐electron bonds between a transition metal and beryllium.

     

Reactions of the Dirhenium(II) Complexes Re2X4(μ-dppm)2 (X=Cl, Br; dppm=Ph2PCH2PPh2) with Isocyanides. XXII. A Comparison of Mixed Carbonyl–Isocyanide Complexes with Those Formed by Re2Cl4(μ-dcpm)2 (dcpm=Cy2PCH2PCy2)

The reactions of the electron-rich triply bonded dirhenium(II) complex Re₂Cl₄(-dcpm)₂ (dcpm=Cy₂PCH₂PCy₂) with the isocyanide ligands XylNC (Xyl=2,6-dimethylphenyl) and t-BuNC afford the complexes Re₂Cl₄(-dcpm)₂(CNXyl) and Re₂Cl₄(-dcpm)₂(CN-t-Bu)₂ which in turn react with CO to give salts of the [Re₂Cl₃(-dcpm)₂(CO)₂(CNXyl)]⁺ and [Re₂Cl₃(-dcpm)₂(CN-t-Bu)₂(CO)]⁺ cations which exist in different isomeric forms. This chemistry is compared with that developed previously for the analogous complexes derived from Re₂Cl₄(-dppm)₂.     

Functionalized cyclopalladated compounds with bidentate Group 15 donor atom ligands: the crystal and molecular structures of [{Pd[5-(COH)C6H3C(H)NCy-C2,N](Cl)}2(μ-Ph2PRPPh2)] (R=CH2CH2, Fe(C5H4)2], [Pd{5-(COH)C6H3C(H)NCy-C2,N}(Ph2PCH2PPh2-P,P)][PF6] and [Pd{5-(COH)C6H3C(H)N(Cy)-C2,N}(Ph2PCH2CH2AsPh2-P,As)][PF6]

The chloro-bridged dinuclear compound [{Pd[5-(COH)C₆H₃C(H)N(Cy)-C2,N]}(μ-Cl)]₂ (1), reacts with tertiary diphosphines in 1:1 molar ratio to give [{Pd[5-(COH)C₆H₃C(H)NCy-C2,N](Cl)}₂(μ-Ph₂PRPPh₂)] (R: CH₂, 2; CH₂CH₂, 3; (CH₂)₄, 4; (CH₂)₆, 5; Fe(C₅H₄)₂, 6; trans-CHCH, 7; C≡C, 8). Treatment of 1 with Ph₂PCH₂CH₂AsPh₂ (arphos) gives the dinuclear complex [{Pd[5-(COH)C₆H₃C(H)N(Cy)-C2,N](Cl)}₂(μ-Ph₂PCH₂CH₂AsPh₂)] (9). The reaction of 1 with tertiary diphosphines or arphos in 1:2 molar ratio in the presence of NH₄PF₆ yields the mononuclear compounds [Pd{5-(COH)C₆H₃C(H)NCy-C2,N}(Ph₂PRPPh₂-P,P)][PF₆] (R: (CH₂)₄, 10; (CH₂)₆, 11; Fe(C₅H₄)₂, 12; 1,2-C₆H₄, 13; cis-CHCH, 14; NH, 15) and [Pd{5-(COH)C₆H₃C(H)N(Cy)-C2,N}(Ph₂PCH₂CH₂AsPh₂-P,As)][PF₆] (16). ¹H-, ³¹P-{¹H}- and ¹³C-{¹H}-NMR, IR and mass spectroscopic data are given. The crystal structures of compounds 3, 6, 9 and 16 have been determined by X-ray crystallography.