抗流感病毒抑制剂Nucleozin衍生物的设计合成及抗病毒活性评价

小分子化合物Nucleozin作为靶向流感病毒核蛋白的抑制剂具有良好的抑制活性。本文围绕Nucleozin分子中与哌嗪环直接相连的芳环部分进行研究。通过钯催化偶联反应合成了一系列Nucleozin衍生物,通过检测所合成化合物对流感病毒H1N1的抑制活性,明确了Nucleozin分子中该部分的构效关系。利用甲基在药物分子设计中的作用,设计将分子中的氯原子替换为甲基,发现与原型分子Nucleozin相比其抑制活性有了明显的提高。本文的结果对该类分子成药性的提高具有积极意义。     

丁烷型木脂素及衍生物的合成和生物活性

报道了一条合成丁烷木脂素的新路线. 以芳香醛为起始原料, Stobbe缩合和烷基化反应为关键步骤, 构建了木脂素骨架, 再经拆分及还原, 可得到相应的苏式和赤式异构体. 经官能团转化得到5个丁烷木脂素和8个丁醚木脂素, 其中3个天然产物为首次合成. 对合成产物进行抗HIV病毒和和抗疱疹病毒活性研究, 部分化合物显示出较高的抗病毒活性, 而且骨架构型对活性影响较大.     

鸭瘟病毒DNA合成动态与部位的电子显微镜放射自显影研究

本文应用~3H-TdR掺入的电子显微镜放射自显影技术研究了鸭瘟病毒DNA在细胞内合成的动态与部位,证明鸭瘟病毒DNA合成的持续时间很长,当病毒核壳体大量装配、病毒粒子已经成熟与释放时,病毒DNA的合成仍继续进行。病毒DNA是在核内电子密度低的基质中合成的。核内毒浆是病毒DNA积聚与核壳体装配的场所,不是病毒DNA合成的部位。     

合成多肽疫苗研究进展

综述了合成多肽疫苗的研究进展,系统地介绍了目前寻找抗原肽的各种方法以及各类可用作疫苗接种的合成多肽抗原的制备和应用.     

黄瓜花叶病毒单克隆抗体的制备及其对株系特异性的研究

黄瓜花叶病毒CMV-Ca分离株免疫的BALB/c小鼠,取脾淋巴细胞与小鼠骨髓瘤SP2/0细胞融合,筛选分离出杂交瘤细胞3A2,回接小鼠所得到的腹水中单克隆抗体,经酶联免疫吸附试验(ELISA)测得效价为1:2.5×10~6。在琼脂双扩散及免疫电泳反应中与同源抗原CMV-Ca产生沉淀线,与黄瓜花叶病毒其它分离株CMV-p,CMV-Cul,CMV-m不起反应。在ELISA试验中与同源抗原反应强烈,只有在病毒浓度很高时才与其他几个分离株起反应,但反应很弱。最后讨论了株系变异可能是部分改变了抗原决定簇的组份或结构以及单克隆抗体用作探针检查这些变异的可能性。     

重金属镉离子人工合成抗原的荧光特性分析

以1-(4-异硫氰苄基)乙烯基二胺-N,N,N′,N′-四乙酸为双功能螯合剂,偶联Cd2+与血清蛋白质分子, 合成了Cd2+人工抗原.利用荧光光谱分析了Cd2+人工合成抗原的荧光特性并得到合成反应的偶联比为11∶ 1～16∶ 1;通过荧光相图分析表明, Cd2+人工抗原合成过程中血清蛋白质的折叠状态符合"二态模型";偏振荧光光谱检测结果显示, Cd2+人工抗原合成中血清蛋白质色氨酸残基的微区构象发生一定变化,导致载体蛋白质构象改变.     

艾滋病（AIDS）病毒抗体检测方法研究进展

对艾滋病(A IDS) 病毒抗体检测方法的研究进展作了较系统的评述。从抗原试剂的构成及样品的来源等方面对各种检测方法进行归纳分类。论述了病毒抗体检测方法研究发展的三个阶段, 分析比较了各种艾滋病毒抗体检测方法的优缺点。给出125 篇参考文献。     

HIV-1 p24抗原电化学免疫传感器

HIV-1的p24抗原是由人类免疫缺陷病毒gag基因编码病毒的主要核心蛋白,艾滋病毒感染的早期诊断主要依赖于准确和灵敏的p24抗原的ELISA法检测~([1]).然而,ELISA法仍然昂贵、费时耗力.在此,我们尝试使用电化学免疫传感器方法快速检测p24抗原,取得了较好的效果,并将有效地降低分析测试的成本.     

禽流感病毒流式微球量子点探针免疫诊断新方法

采用微波法水相中合成羧基化的绿色量子点,通过羧基与禽流感单克隆抗体氨基的共价结合,制备了检测禽流感病毒的探针,并结合流式微球技术,建立了量子点生物探针流式微球免疫检测禽流感病毒的新方法.以聚苯乙烯微球为蛋白质载体,将多克隆抗体包被到荧光微球上,依次加入待测抗原和量子点生物探针,形成双抗体夹心复合物,用流式细胞仪进行检测.实验结果表明,多抗和单抗的最佳质量浓度分别为92和4 mg/L,检测禽流感病毒比双抗体夹心ELISA灵敏16倍,比FITC标记单抗检测方法灵敏4倍.对阳性尿囊液的检测与ELISA呈现良好的相关性,不与鸡传染性支气管炎病毒、鸡马力克氏病毒、新城疫病毒等发生交叉反应.     

克隆的乙型肝炎病毒核心抗原基因的起始密码上游序列对表达的影响

本文报道以双脱氧链终止法测定了乙型肝炎病毒(HBV)adw亚型不同表达水平克隆株中核心抗原基因(HBc)的核苷酸序列。结果表明,表达水平的差异主要是由于核心抗原基因起始密码上游核苷酸序列结构的不同所致。低表达克隆株中的HBc在起始密码前形成一个茎状的二级结构;高表达克隆株则在Bal-31外切酶的作用下,此结构被完金切除;中庋表达克隆株此结构的一半被切除。显然,二级结构的存在阻碍了核心抗原基因的转录和转译。从而降低了核心抗原蛋白在大肠杆菌中的合成量。     

2-(N-三氟乙酰-N-三甲基硅烷基)氨基-3-三甲基硅烷氧基-羧酸三甲基硅烷酯和2-(N-三氟乙酰)-2,3-脱氢氨基酸的合成

利用三氟甲基磺酸三甲基硅烷酯, 我们合成了一种新的、化学活性很高的合成中间产物2-(N-三氟乙酰-N-三甲基硅烷基)氨基-1, 1-二(三甲基硅烷氧基)乙烯。脂肪醛或芳香醛发生碳碳成键的加成反应, 生成β碳原子上带有易离去基团三甲基硅烷氧基、N原子上带有保护基团三氟乙酰基的α氨基酸三甲基硅烷酯。消除反应得到了一个合成α、β脱氢氨基酸的可行途径。这类化合物是合成复杂多肽和肽生物碱的基元物。     

Peptide synthesis using pentafluorophenyl diphenylphosphate (FDP) as coupling reagent

Comparison of the reaction speed and yield with its analogues and some conventional peptide coupling reagents, pentafluorophenyl diphenylphosphate (FDP) was shown to be a more preferable "active ester" type reagent for the peptide synthesis. The synthesis of oligopeptides using FDP was achieved with high yields. The influences of several reaction parameters such as solvent, base, additive and temperature on the coupling reaction were studied using HPLC method. The degree of racemization with FDP determined by HPLC or Young test was shown to be lower than that of DCCI. Octapeptide Gly-Cys(Bzl)-Ser-Gly-Lys-Leu-Ile-Cys(Bzl)-OH, corresponding to the amino acid sequence of gp41 of HIV-1, was successfully synthesized by 5+3 approach using FDP with high yield.     

A bioorganometallic approach for the electrochemical detection of proteins: a study on the interaction of ferrocene-peptide conjugates with papain in solution and on Au surfaces

In this paper, a new bioorganometallic approach for the detection of proteins using surface-bound ferrocene-peptide conjugates is presented. Specifically, a series of peptide conjugates of 1'-aminoferrocene-1-carboxylic acid (ferrocene amino acid, Fca) is synthesized: Boc-Fca-Gly-Gly-Tyr(Bzl)-Arg(NO2)-OMe (2), Thc-Fca-Gly-Gly-Tyr(Bzl)-Arg(NO2)-OMe (3), Thc-Fca-Gly-Gly-Tyr(Bzl)-Arg(NO2)-OH (4), Boc-Fca-Gly-Gly-Arg(Mtr)-Tyr-OMe (7), Thc-Fca-Gly-Gly-Arg(Mtr)-Tyr-OMe (8), Thc-Fca-Gly-Gly-Arg(Mtr)-Tyr-OH (9), Thc-Fca-Gly-Gly-Arg-Tyr-OH (10). The peptide is conjugated to the C-terminal side of Fca and compounds 4, 7-10 possess a thiostic acid linked to the N-terminal side of Fca in order to facilitate formation of thin films on gold substrates. Competitive inhibition towards papain was determined for Thc-Fca-Gly-Gly-Tyr(Bzl)-Arg(NO2)-OH (4), Thc-Fca-Gly-Gly-Arg(Mtr)-Tyr-OH (9) and Thc-Fca-Gly-Gly-Arg-Tyr-OH (10). The binding interaction between the peptide modified substrates and papain enzyme was studied using real-time surface plasmon resonance (SPR) imaging. Peptide 10 showed the strongest binding affinity to papain. Adsorption/desorption rate constants were ka = 1.75+/-0.05 x 10(5) M(-1) s(-1) and kd = 2.90 +/- 0.05 x 10(-2) s(-1). Interactions of papain with Fca-peptide 10 were investigated by cyclic voltammetry. The interaction results were also verified by measuring the electrochemical response of the peptide-papain interaction as function of increasing enzyme concentration. A linear relationship was observed for papain concentration of up to 80 nM. Shifting to higher potentials as well as decrease in the overall signal intensity was observed. The detection limit was 4 x 10(-9) M.     

2-Benzoylpyridine thiosemicarbazone as a novel reagent for the single pot synthesis of dinuclear Cu(I)-Cu(II) complexes: formation of stable copper(II)-iodide bonds

2-Benzoylpyridine thiosemicarbazone {R(1)R(2)C(2)=N(2)·N(3)H-C(1)(=S)-N(4)H(2), R(1) = py-N(1), R(2) = Ph; Hbpytsc} with copper(I) iodide in acetonitrile-dichloromethane mixture has formed stable Cu(II)-I bonds in a dark green Cu(II) iodo-bridged dimer, [Cu(2)(II)(μ-I)(2)(η(3)-N(1),N(2),S-bpytsc)(2)] 1. Copper(I) bromide also formed similar Cu(II)-Br bonds in a dark green Cu(II) bromo-bridged dimer, [Cu(2)(II)(μ-Br)(2)(η(3)-N(1),N(2),S-bpytsc)(2)] 3. The formation of dimers 1 and 3 appears to be due to a proton coupled electron transfer (PCET) process wherein copper(I) loses an electron to form copper(II), and this is accompanied by a loss of -N(3)H proton of Hbpytsc ligand resulting in the formation of anionic bpytsc(-). When copper(I) iodide was reacted with triphenylphosphine (PPh(3)) in acetonitrile followed by the addition of 2-benzoylpyridine thiosemicarbazone in dichloromethane (Cu?:?PPh(3)?:?Hbpytsc in the molar ratio 1:1:1), both Cu(II) dimer 1 and an orange Cu(I) sulfur-bridged dimer, [Cu(2)(I)I(2)(μ-S-Hbpytsc)(2)(PPh(3))(2)] 2 were formed. Copper(I) bromide with PPh(3) and Hbpytsc also formed Cu(II) dimer 3 and an orange Cu(I) sulfur-bridged dimer, [Cu(2)(I)Br(2)(μ-S-Hbpytsc)(2)(PPh(3))(2)] 4. While complexes 2 and 4 exist as sulfur-bridged Cu(I) dimers, 1 and 3 are halogen-bridged. The central Cu(2)S(2) cores of 2 and 4 as well as Cu(2)X(2) of 1 (X = I) and 3 (X = Br) are parallelograms. One set of Cu(II)-I and Cu(II)-Br bonds are short, while the second set is very long {1, Cu-I, 2.565(1), 3.313(1) ?; 3, Cu-Br, 2.391(1), 3.111(1) ?}. The Cu···Cu separations are long in all four complexes {1, 4.126(1); 2, 3.857(1); 3, 3.227(1); 4, 3.285(1) ?}, more than twice the van der Waals radius of a Cu atom, 2.80 ?. The pyridyl group appears to be necessary for stabilizing the Cu(II)-I bond, as this group can accept π-electrons from the metal.     

From the tetra(amino) phosphonium cation, [P(NHPh)4]+, to the tetra(imino) phosphate trianion, [P(NPh)4]3-, two-faced ligands that bind anions and cations

The tetraanilino phosphonium cation, [P(N(H)Ph)4]+, 1+, is sequentially deprotonated by Bu(n)Li in thf. The deprotonation reaction of the chloride derivative, Cl, was monitored by (31)P NMR, which revealed the successive formation of the neutral [P(N(H)Ph)3(NPh)], 2, the monoanionic [P(N(H)Ph)2(NPh)2]-, 3-, the dianionic [P(N(H)Ph)(NPh)3]2-, 4(2-), and finally the trianionic species [P(NPh)(4)](3-), (3-). Considering the isoelectronic relationship of oxo, =O, and imino groups, =NR, as well as hydroxy, -OH, and amino groups, -N(H)R, the neutral complex corresponds to phosphoric acid, H3PO4, whereas the anions 3-, 4(2-) and 5(3-) are analogues of dihydrogen phosphate, H2PO4-, monohydrogenphosphate, HPO4(2-), and orthophosphate ions, PO4(3-), respectively. Solid state structures were obtained of 1Cl, 2LiCl(thf)(2), 3Li(thf)(3.5), 3Li(2)Cl(thf)(4.25), 3Li(2)Cl(thf)(6) and 5Li(4)Cl(thf)(4). All systems provide two separate N-P-N chelation sites at opposite ligand faces, either consisting of the di(amino) arrangement P(NH)(2), acting as a double H-bond donor, the di(imino) arrangement PN(2), donating two electron pairs, or the mixed amino imino arrangement P(N)(NH), which supplies both electron pair and H-donor site. Interesting in this aspect is the mixed amino imino derivative 3- which has the ability to chelate a Lewis acid, such as a metal ion, at one face and a Lewis base, such as an anionic or neutral donor at the opposite ligand face. The formation of 1-D aggregates and the entrapment of lithium chloride are key characteristics of the supramolecular structures of the discussed complexes.     

Cationic and neutral diphenyldiazomethanerhodium(I) complexes as catalytically active species in the C-C coupling reaction of olefins and diphenyldiazomethane

Cationic rhodium(I) complexes cis-[Rh(acetone)2(L)(L')]+ (2: L = L'=C8H14; 3: L=C8H14; L'=PiPr3; 4: L=L'=PiPr3), prepared from [RhCl(C8H14)2]2] and isolated as PF6 salts, catalyze the C-C coupling reaction of diphenyldiazomethane with ethene, propene, and styrene. In most cases, a mixture of isomeric olefins and cyclopropanes were obtained which are formally built up by one equivalent of RCH=CH2 (R = H, Me, Ph) and one equivalent of CPh2. The efficiency and selectivity of the catalyst depends significantly on the coordination sphere around the rhodium(I) center. Treatment of 4 with Ph2CN2 in the molar ratio of 1:1 and 1:2 gave the complexes trans-[Rh(PiPr3)2(acetone)(eta1-N2CPh2)]PF6 (8) and trans-[Rh(PiPr3)2(eta1-N2CPh2)2]PF6 (9), of which 8 was characterized by X-ray crystallography. Since 8 and 9 not only react with ethene but also catalyze the reaction of C2H4 and free Ph2CN2, they can be regarded as intermediates (possibly resting states) in the C-C coupling process. The lability of 8 and 9 is illustrated by the reactions with pyridine and NaX (X=Cl, Br, I, N3) which afford the mono(diphenyldiazomethane)rhodium(I) compounds trans-[Rh(PiPr3)2(py)(eta1-N2CPh2)]PF6 (10) and trans-[RhX(eta1-N2CPh2)(PiPr3)2] (11-14), respectively. The catalytic activity of the neutral complexes 11 - 14 is somewhat less than that of the cationic species 8, 9 and decreases in the order Cl > Br> I > N3.     

Amino acid and peptide bioconjugates of copper(II) and zinc(II) complexes with a modified N,N-bis(2-picolyl)amine ligand

Four chelating nitrogen ligands 2-5 derived from N,N-bis(2-picolyl)amine (bpa, 1) were synthesized, namely, (PyCH(2))(2)N-CH(2)-p-C(6)H(4)-CO(2)R (R = Me, 2, and R = H, 3) and (PyCH(2))(2)N-(CH(2))(n)-CO(2)H (n = 2, 4, and n = 5, 5). Amino acid conjugates 6 and 7 were formed by condensation of 3 with H-Phe-OMe and H-betaAla-OMe, respectively. Cu(II) and Zn(II) complexes of 1-7 were prepared and fully characterized. The X-ray structures of 1(Zn), 2(Zn), 4(Cu), and 7(Cu) were determined. The Zn complexes 1(Zn) and 2(Zn) as well as 7(Cu) show a distorted trigonal bipyramidal coordination environment in the solid state. An octahedral complex is observed for 4(Cu) which forms chains along the crystallographic b axis by intermolecular coordination of the carboxylic acid to the metal ion of a neighboring complex. Ligand 3 was used to prepare the peptide bioconjugate 8 (3-Ahx-Pro-Lys-Lys-Lys-Arg-Lys-Phe-NH(2)) with a nuclear localization signal (nls) heptapeptide by solid phase synthesis. Cu(II) and Zn(II) complexes of 8 were synthesized in situ and studied by FAB-MS, ESI-MS, UV/vis, and EPR (for 8(Cu)), and FAB-MS, ESI-MS, and NMR (for 8(Zn)). All spectroscopic results clearly support metal coordination to the bpa ligand in the bioconjugates 8(M), even in the presence of other potential ligands from amino acid side chains of the peptide. We suggest metal-peptide conjugates like 8(M) as artificial metallochaperones because they have the potential to deliver metal ions to specific compartments in the cell as determined by the peptide moieties.     

抗菌肽IB-367的固相合成与抑菌活性

采用固相合成方法,以Rink Amide树脂为载体,Fmoc保护氨基酸为原料,经苯并三唑-1-四甲基六氟磷酸酯(HBTU)/N,N-二异丙基乙胺(DIEA)缩合,三氟乙酸/苯甲硫醚/乙二硫醇/苯甲醚裂解体系脱除保护基制得IB-367线性肽(4); 4经双氧水氧化制得IB-367一环肽(5); 5经碘乙醇溶液氧化合成抗菌肽IB-367(6),收率34.1%,纯度＞95.0%,其结构经MS(ESI)和氨基酸组成分析确证。抑菌活性研究结果表明：6对大肠杆菌和金黄色葡萄球菌的最小抑菌浓度为5.0 μg·mL^-1。     

Cube-type nitrido complexes containing titanium and zinc/copper

Several azaheterometallocubane complexes containing [MTi3N4] cores have been prepared by the reaction of [{Ti(eta5-C5Me5)(mu-NH)}3(mu3-N)] (1) with zinc(II) and copper(I) derivatives. The treatment of 1 with zinc dichloride in toluene at room temperature produces the adduct [Cl2Zn{(mu3-NH)3Ti3(eta5-C5Me5)3(mu3-N)}] (2). Attempts to crystallize 2 in dichloromethane gave yellow crystals of the ammonia adduct [(H3N)Cl2Zn{(mu3-NH)Ti3(eta5-C5Me5)3(mu-NH)2(mu3-N)}] (3). The analogous reaction of 1 with alkyl, (trimethylsilyl)cyclopentadienyl, or amido zinc complexes [ZnR2] leads to the cube-type derivatives [RZn{(mu3-N)(mu3-NH)2Ti3(eta5-C5Me5)3(mu3-N)}] (R = CH2SiMe3 (5), CH2Ph (6), Me (7), C5H4SiMe3 (8), N(SiMe3)2 (9)) via RH elimination. The amido complex 9 decomposes in the presence of ambient light to generate the alkyl derivative [{Me3Si(H)N(Me)2SiCH2}Zn{(mu3-N)(mu3-NH)2Ti3(eta5-C5Me5)3(mu3-N)}] (10). The chloride complex 2 reacts with lithium cyclopentadienyl or lithium indenyl reagents to give the cyclopentadienyl or indenyl zinc derivatives [RZn{(mu3-N)(mu3-NH)2Ti3(eta5-C5Me5)3(mu3-N)}] (R = C5H5 (11), C9H7 (12)). Treatment of 1 with copper(I) halides in toluene at room temperature leads to the adducts [XCu{(mu3-NH)3Ti3(eta5-C5Me5)3(mu3-N)}] (X = Cl (13), I (14)). Complex 13 reacts with lithium bis(trimethylsilyl)amido in toluene to give the precipitation of [{Cu(mu4-N)(mu3-NH)2Ti3(eta5-C5Me5)3(mu3-N)}2] (15). Complex 15 is prepared in a higher yield through the reaction of 1 with [{CuN(SiMe3)2}4] in toluene at 150 degrees C. The addition of triphenylphosphane to 15 in toluene produces the single-cube compound [(Ph3P)Cu{(mu3-N)(mu3-NH)2Ti3(eta5-C5Me5)3(mu3-N)}] (16). The X-ray crystal structures of 3, 8, 9, and 15 have been determined.     

Synthesis of CuII-RuII-CuII trinuclear complexes via redox reaction of copperI across thiosemicarbazones coordinated to rutheniumII

Pyridine-2-carbaldehyde thiosemicarbazones [C5H4N1-C(H)N2-N3H-C(S)-N4HR, R = H, L1H2; CH3, L2H2-Me; CH2CH3, L3H2-Et] with Ru(PPh3)3Cl2 have formed mononuclear RuII precursors for the generation of trinuclear complexes. The reaction of 2 mol each of L1H2, L2H2-Me, or L3H2-Et with Ru(PPh3)3Cl2 in the presence of Et3N has yielded mononuclear complexes [Ru(N3,S-L1H)2(PPh3)2] (1), [Ru(N3,S-L2H-Me)2(PPh3)2] (2), and [Ru(N3,S-L3H)2(PPh3)2] (3). The addition of 2 equiv of copperI chloride solution to complex 1 in acetonitrile has formed a novel trinuclear complex, (Ph3P)2RuII(L1)2CuII2Cl2 (4), in which the pendant amino group (-N4H2) loses one hydrogen along with the oxidation of CuI to CuII. In this complex, RuII is bonded to two P, two S, and two N3 atoms, while each CuII is coordinated to N1, N2, N4, and Cl atoms. Reaction with copper(I) bromide yielded a similar trinuclear complex, (Ph3P)2Ru(L1)2CuII2Br2 (5). From precursors 2 and 3, analogous complexes (Ph3P)2RuII(L2-Me)2CuII2Cl2 (6), (Ph3P)2RuII(L2-Me)2CuII2Br2 (7), (Ph3P)2RuII(L3-Et)2CuII2Cl2 (8), and (Ph3P)2RuII(L3-Et)2CuII2Br2 (9) have been synthesized. These complexes have been characterized using analytical, spectroscopic, and electrochemical techniques. Single-crystal X-ray crystallography has been carried out for precursor 2 and all of the trinuclear complexes, 4-9. X-band electron spin resonance and UV-vis spectra have confirmed the presence of CuII. The cyclic voltammetry studies support the RuII/RuIII redox behavior of this metal in trinuclear complexes.