4-酰基-5-吡唑酮的二烃基锡配合物的合成、表征及其抗癌活性研究

目的　设计合成一系列新型有机锡化合物,通过药理筛选寻找具有抗肿瘤活性的药物并探讨其构效关系。方法　以4-酰基-5-吡唑酮为配体合成有机锡化合物,用元素分析、红外光谱、 ¹H,¹³C,¹¹⁹Sn NMR等方法对得到的化合物作结构表征并利用几种药理模型进行体外抗癌活性筛选。结果　合成了一系列R₂SnL₂ 型有机锡新配合物并确定其结构。结论　药理筛选结果表明,多个化合物(3～6,9～11)对HL-60, HCT-8, Bel-7402,BGC-823和KB癌细胞有效,显示了较强的抗癌活性。     

孤啡肽及其片段的合成、痛觉调节作用和对免疫功能的影响

目的：研究孤啡肽(NC)与其4个片段(NC(1-15)NH₂, NC(1-13)NH₂, NC(1-11)NH₂, NC(1-5)NH₂) 在痛觉调节和免疫活性上的变化,探讨NC的构效关系。方法：固相多肽合成法合成NC及其片段;甩尾法测定它们对小鼠的痛敏作用和对吗啡镇痛作用的拮抗;T细胞玫瑰花结形成百分率和红细胞免疫粘附能力评测对免疫功能的影响。结果：虽然NC及其片段均有痛敏作用并可拮抗吗啡的镇痛作用,但NC(1-11)NH₂和NC(1-5)NH₂比母体活性约降低100倍,而NC(1-13)NH₂和NC(1-15)NH₂与母体有相同的活性。NC及片段(0.3～3 nmol.kg^-1)对T细胞免疫功能均有促进作用;NC(1-11)NH₂(0.3 nmol.kg^-1)对红细胞的免疫粘附能力有促进作用;NC(1-5)NH₂(0.3～30 nmol.kg^-1)不影响红细胞的免疫功能。结论：C端在NC的构效关系中有重要的作用。     

双取代乙二胺金属配合物的合成及抗菌活性

目的　设计合成一系列双取代乙二胺过渡金属配合物并寻找抗菌活性药物。方法　以N ,N′-双取代乙二胺为配体合成过渡金属配合物,并进行体外抑菌活性筛选。结果　合成了6个(Ⅲ_1-6)新配合物,其结构经红外光谱、元素分析确证。结论　初步抑菌实验表明,合成的配合物对多种菌株有明显的抑制活性,有进一步研究的价值。     

［Ln(Phen)2(5-Fu)3(NO3)］(NO3)2的合成、表征及体外抗肿瘤活性研究

目的研究稀土元素的生物化学作用和无机、有机抗肿瘤药物的协同作用。方法用稀土硝酸盐、邻菲罗啉(Phen)、氟尿嘧啶(5-Fu)合成了7种稀土配合物。用MTT,SRB法进行体外抗肿瘤活性研究。结果经过元素分析、摩尔电导率测定、差热分析、红外光谱测定和核磁共振谱测定，初步确定配合物化学式为［Ln(Phen)₂(5-Fu)₃(NO₃)］(NO₃)₂。配合物对K562(人粒细胞白血病细胞株)、CA(人肝癌细胞株)、SGC-7901(人胃癌细胞株)、HCT(人结肠癌细胞株)、GLC-82(人肺癌细胞株)和CNE(人鼻咽癌细胞株)增殖有很强的抑制作用。结论配合物产生了抗肿瘤协同作用。     

1,3-二甲基-4-酰基-5-吡唑酮二烃基锡抗癌配合物与单核苷酸作用的研究

目的研究1,3-二甲基-4-乙酰基-5-吡唑酮(HL₁)的二烃基锡抗癌配合物在近生理条件下与单核苷酸和DNA的作用机制。方法以高分辨核磁共振研究有机锡化合物在不同时间、不同条件下与核苷酸的作用方式并用UV法进一步研究有机锡化合物与DNA的作用。 结果(L₁)₂SnEt₂与AMP作用时可引起AMP碱基上H(8),H(2)和³¹P峰化学位移值显著变化并可引起DNA增色效应;(L₁)₂SnMe₂与AMP作用时只引起AMP的³¹P化学位移值的显著变化,未观察到AMP的H(8)和H(2)化学位移值的明显改变,(L₁)₂SnMe₂与DNA的作用可导致DNA的减色效应;(L₁)₂SnEt₂和(L₁)₂SnMe₂在与dCMP和dGMP作用时只引起dCMP和dGMP的³¹P的化学位移数值的明显变化。结论 3-二甲基-4-乙酰基-5-吡唑酮的二乙基锡配合物(L₁)₂SnEt₂可与AMP的碱基N₁和磷酸根氧原子螯合并可能会破坏DNA的双股螺旋结构;而二甲基锡配合物(L₁)₂SnMe₂易与单核苷酸的磷酸氧结合,难以与单核苷酸的碱基氮原子稳定结合且只引起DNA双股螺旋的收缩。     

有机锡化合物抗肿瘤生物活性研究

有机锡(IV)化合物能明显地抑制大鼠脑组织中PKC活性和肿瘤细胞增殖，且两者之间存在着相关性。抗肿瘤活性的构效关系是：(1)有机基团R决定整个化合物的生物活性，Ph>Et>n-Bu；(2)卤素的电负性影响化合物活性的大小。抗肿瘤活性可能通过[snR₂]²⁺实现。并能部分地阻断HL-60细胞周期中的G_I期向S期的移行。[SnPh₂F₂]，[SnPh₂(CysOS)]H₂O和[SnPh₂Cl2·phen(CH₃)₂]对PKC的IC₅₀值分别为25，15和20 umol·L^-1，对HL-60细胞的IC₅₀值分别为0．5，4．0和0．3umol·L^-1。但它们无诱导分化HL-60和K562细胞的作用。     

国家食品药品监督管理局国家药品标准修订——法莫替丁钙镁咀嚼片

本品每片含法莫替丁(C₈H₁₅N₇O₂S₃)、碳酸钙(CaCO₃)与氢氧化镁[Mg(OH)₂]均应为标示量的90.0%~110.0%。     

水杨醛缩氨基脲含硅有机锡化合物的合成和抑菌活性测试

目的 合成新型的含硅有机锡化合物,并进行抑菌活性测试。方法 利用4种水杨醛缩氨基脲配体与(Me₃SiCH₂)₂SnCl₂通过常温下的自组装反应合成了4种新型含硅有机锡化合物,利用红外光谱、核磁共振以及X-射线单晶衍射对所得产物的结构进行表征,通过纸片扩散法测试产物的抑菌活性。结果 在化合物1~4中,水杨醛缩氨基脲配体均呈现三齿配体形式与锡原子螯合配位,中心锡原子呈五配位的三角双锥结构。抑菌实验发现3种新合成的水杨醛缩氨基脲含硅有机锡化合物对G⁺菌包括金黄色葡萄球菌、G⁺蜡样芽孢杆菌和真菌蜡蚧轮枝孢菌显示抑菌作用。化合物3对金黄色葡萄球菌的抑菌圈为20 mm,最低抑菌浓度为25.17 mol·L^-1。此类化合物对G^-菌包括大肠杆菌、伤寒杆菌、产气肠杆菌无明显抑制作用。结论 水杨醛缩氨基脲含硅有机锡化合物对G⁺菌包括金黄色葡萄球菌及真菌蜡蚧轮枝孢菌具有一定的抑菌活性,对G^-菌无明显的抑制作用。     

柿子黄酮类化合物的双水相系统纯化及止血凝血抗炎作用研究

目的 采用聚乙二醇600(PEG 600)-(NH4)2SO4的双水相体系纯化柿子黄酮(flavonoids extracts from persimmon,FEP)并考察其止血凝血抗炎作用。方法 以FEP得率为指标,采用单因素试验设计,探讨PEG 600-(NH₄)₂SO₄双水相体系中的PEG 600质量分数、(NH₄)₂SO₄质量分数、温度、MgCl₂用量、pH对FEP提取率的影响,并采用Box-Behnken中心组合方法设计3因素3水平的响应面法试验,选择Design-Expert 8.05软件进行响应面分析。以KM小鼠为试验对象,采用剪尾法、毛细管法、二甲苯致炎法对FEP进行止血、凝血、急性抗炎作用的初步研究,并分别与三七粉和阿司匹林进行比较。结果 最佳纯化条件：PEG 600质量分数为12%,(NH₄)₂SO₄质量分数为28%,pH值为8,最终得率为95.97%。FEP可使小鼠的出血时间和凝血时间明显缩短,对二甲苯所致的小鼠耳肿胀具有较显著的抑制作用。结论 本法所形成的双水相体系,操作简便,高效温和并有效保持生物活性;可以考虑将FEP作为活性成分添加入口腔用生活用品或药品。     

4-氨基-3-(α-呋喃)-5-巯基-1,2,4-三唑的SCHIFF氏碱、铜(Ⅱ)配合物的杀菌、抑霉活性

为筛选新型杀菌抑霉药物,本文报道了标题化合物对7种细菌和4种真菌的杀菌抑霉活性。18～64%的受试菌株对配体Ⅰ₂～Ⅰ₁₁和配合物Ⅱ₂～Ⅱ₁₁表现敏感;多数化合物对大肠杆菌有抑制作用;串珠镰刀菌、毛霉菌、黄曲霉菌、青霉菌对化合物Ⅰ₂高度敏感,抑菌圈直径为15～25mm;化合物Ⅰ₁₁和Ⅱ₁₁分别对73%和64%的菌株有抑菌作用。     

Synthesis,biological activity and conformational analysis of cyclic GRF analogs

A novel cyclic GRF analog, cyclo(Asp⁸-Lys¹²)-[Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂, i.e. cyclo^8.12[Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂, was synthesized by the solid phase procedure and found to retain significant biological activity. Solid phase cyclization of Asp⁸ to Lys¹² proceeded rapidly (～2h) using the BOP reagent. Substitution of Ala¹² with d -Ala² and/or NH₂-terminal replacement (desNH₂-Tyr¹ or N-MeTyr¹) in the cyclo^8.12[Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂ system resulted in highly potent analogs that were also active in vivo. Conformational analysis (circular dichroism and molecular dynamics calculations based on NOE-derived distance constraints) demonstrated that cyclo^8.12[Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂ contains a long α-helical segment even in aqueous solution. A series of cyclo^8.12 stereoisomers containing d -Asp⁸ and/or d -Lys¹² were prepared and also found to be highly potent and to retain significant α-helical conformation. The high biological activity of cyclo^8.12[N-MeTyr¹,d -Ala²,Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂ may be explained on the basis of retention of a preferred bioactive conformation.     

Study of the cytotoxicity and antioxidant capacity of N/OFQ(1–13)NH2 and its structural analogues

The effect of side-chain shortening of N/OFQ(1–13)NH₂ at position 9 ([Orn⁹]N/OFQ(1–13)NH₂, [Dab⁹]N/OFQ(1–13)NH₂, [Dap⁹]N/OFQ(1–13)NH₂) was studied regarding potential toxicity and antioxidant capacity. Staurosporine- and H2O2-induced damage of SH-SY5Y neuroblastoma cells was not changed in the presence of N/OFQ(1–13)NH2 and [Orn⁹]N/OFQ(1–13)NH₂, but was strongly enhanced in the presence of [Dab⁹]N/OFQ(1–13)NH₂ and [Dap⁹]N/OFQ(1–13)NH₂. Moreover, treatment of cells with the latter two analogues alone led to cell injury. Neuropeptide-dependent differences in the viability of SH-SY5Y cells were also observed, i.e., a cytoprotective effect was observed only in the presence of N/OFQ(1–13)NH₂ and [Orn⁹]N/OFQ(1–13)NH₂. Compared to [Dab⁹]N/OFQ(1–13)NH₂ and [Dap⁹]N/OFQ(1–13)NH₂, the effects of N/OFQ(1–13)NH₂ and [Orn⁹]N/OFQ(1–13)NH₂ were more beneficial in systems generating free oxygen radicals (O2^? and OH), as well as on the antioxidant status of rat brain and liver. Taken together, our findings show that N/OFQ(1–13)NH₂ and its structural analogue [Orn⁹]N/OFQ(1–13)NH₂ possess more favorable profiles than the other two nociceptin (N/OFQ) analogues. The present results suggest that shortening of the side-chain of N/OFQ(1–13)NH2 might increase cell damage and reduce the viability of SH-SY5Y neuroblastoma cells. Moreover, such alterations may lead to changes in free-oxygen generating systems and in antioxidant status in animal tissues.     

In vitro pharmacological characterisation of a novel cyclic nociceptin/orphanin FQ analogue c[Cys<Superscript>7,10</Superscript>]N/OFQ(1–13)NH<Subscript>2</Subscript>

Nociceptin/orphanin FQ (N/OFQ) is the endogenous 17 amino acid peptide ligand for the G_i-protein-coupled N/OFQ receptor (NOP). In an attempt to improve the metabolic stability of N/OFQ, we have produced a truncated cyclic analogue with cysteine residues at positions 7 and 10, c[Cys^7,10]N/OFQ(1–13)NH₂ (c[Cys^7,10]). c[Cys^7,10], the template N/OFQ(1–13)NH₂ and N/OFQ displaced the binding of [³H]N/OFQ to Chinese hamster ovary cells expressing recombinant human NOP (CHO_hNOP) with pK _i values of 9.98, 9.83 and 9.18, respectively. In addition, c[Cys^7,10], N/OFQ(1–13)NH₂ and N/OFQ stimulated the binding of guanosine triphosphate gamma [³⁵S] to CHO_hNOP cells with pEC₅₀/E _max (stimulation factor) of 9.16/5.5, 9.11/4.9 and 8.35/5.5, respectively. c[Cys^7,10], N/OFQ(1–13)NH₂ and N/OFQ inhibited forskolin-stimulated cyclic adenosine monophosphate (cAMP) formation with pEC₅₀ values of 10.08, 10.11 and 9.78, respectively. All ligands produced complete inhibition of cAMP formation. In both functional assays, c[Cys^7,10] was a full agonist. In a series of metabolism experiments, incubation of 1 nM c[Cys^7,10], N/OFQ(1–13)NH₂ and N/OFQ with a rat brain homogenate produced a time-dependent loss of peptide that was greatest for the native peptide N/OFQ. Amidation in N/OFQ(1–13)NH₂ produced some metabolic protection, but this was not significantly improved by further inclusion of c[Cys^7,10]. In summary, c[Cys^7,10] is a high-affinity, high-potency full agonist of the NOP receptor. However, we were unable to demonstrate clear metabolic protection.     

Degradation of growth hormone releasing factor analogs in neutral aqueous solution is related to deamidation of asparagine residues

The incubation of a solution of the human growth hormone releasing factor analog, [Leu²⁷] hGRF(1-32)NH₂ at pH 7.4 and 37°, resulted in extensive degradation of the sample. The major degradation products were identified as the peptides [β-Asp⁸, Leu²⁷] hGRF(l-32)NH₂ and [α-Asp⁸, Leu²⁷] hGRF(1-32)NH₂, produced by deamidation of the Asn⁸ residue. When tested as growth hormone (GH) secretagogues in cultured bovine anterior pituitary cells, [β-Asp⁸, Leu²⁷] hGRF(l-32)NH₂ was estimated to be 400-500 times less potent than the parent Asn⁸ peptide, while [2-Asp⁸, Leu²⁷] hGRF(l-32)NH₂ was calculated to be 25 times less potent than the parent Asn⁸ peptide. Three additional analogs of [Leu²⁷] hGRF(1-32)NH₂ containing either Ser or Asn at positions 8 and 28 were prepared and evaluated for their GH releasing activity and stability in aqueous phosphate buffer (pH 7.4, 37°). Based on disappearance kinetics, [Leu²⁷] hGRF(1-32)NH₂ had a half-life of 202 h while the other analogs had the following half-lives: [Leu²⁷, Asn²⁸] hGRF(1-32)NH_: (150h); [Ser⁸, Leu²⁷, Asn²⁸] hGRF(l-32)NH₂ (746h); and [Ser⁸, Leu²⁷] hGRF(1-32)NH₂ (1550 h). After 14 days, incubated samples of the Asn⁸ analogs lost GH releasing potency, while the Ser⁸ analogs retained full potency. The potential for loss of biological activity brought about by deamidation of other engineered peptides and proteins should be considered in their design.     

Lipoxygenase inhibitory sphingolipids from Launaea nudicaulis

Four new sphingolipids: nudicaulin A [(2S,3S,4R,14E)-2-{[octadecanoyl]amino}tetraeicos-14-ene-1,3,4-triol; 1], nudicaulin B [(2S,3S,4R,14E)-2-{[(2R)-2-hydroxyoctadecanoyl]amino}tetraeicos-14-ene-1,3,4-triol; 2], nudicaulin C [(2S,3S,4R,14E)-2-{[(2R)-2-hydroxyoctadecanoyl]amino}tetraeicos-14-ene-1,3,4-triol-1-O-β-d-glucopyranoside; 3], and nudicaulin D [(2S,3S,4R)-2-{[(2R,3S,12E)-2,3-dihydroxyeicos-12-enoyl]amino}octadecane-1,3,4-triol; 4] together with 1-hexatriacontanol, β-sitosterol, octadecyl 4-hydroxycinnamate, elaidic acid, cholesta-5,22-diene-3,7-diol, oleanolic acid, apigenin, and β-sitosterol 3-O-β-d-glucopyranoside were isolated from the methanolic extract of the whole plant of Launaea nudicaulis. Their structures were elucidated using ¹H and ¹³C NMR spectra and 2D NMR analyses (HMQC, HMBC, and COSY) in combination with mass spectrometry (EI-MS, HR-EI-MS, FAB-MS, and HR-FAB-MS) experiments and comparison with literature data of related compounds. Compounds 1–4 displayed moderate inhibitory potential against enzyme lipoxygenase in concentration-dependent manner with IC₅₀ value ranges 103–193 μM.     

Platinum and Palladium‐triazole Complexes as Highly Potential Antitumor Agents

The palladium complexes [(dppe)Pd(L)₂PdCl₂], [(dppe)Pd(L)₂PtCl₂], [(dppp)Pd(L)₂PdCl₂], [(dppm) Pd(L)₂NiCl₂], and [(dppm)Pd(L)₂SnCl₄] 15–19 were prepared. The antiproliferative activity of the newly synthesized complexes as well as their previously prepared analogues 3–14 and 20–26 were screened against a large panel of human cancer cell lines derived from haematological CD4⁺ human T‐cells containing an integrated HTLV‐1 genome (MT‐4). The complex 12a , b exhibited remarkable antiproliferative activity against MT‐4, CD4⁺ human acute T‐lymphoblastic leukemia (CCRF‐CEM), human splenic B‐lymphoblastoid cells (WIL‐2NS), human acute B‐lymphoblastic leukemia (CCRF‐SB), skin melanoma (SK‐MEL‐28), and prostate carcinoma (DU145) cell lines (CC₅₀ = 0.5 μM, 0.4 ± 0.05 μM, 0.6 ± 0.05 μM, 0.4 ± 0.1 μM, and 0.8 ± 0.2 μM, respectively), meanwhile, 9a , b , 14a , b , and 23 showed significant activity against the CCRF‐SB cell lines (CC₅₀ = 0.6 ± 0.06 μM, 0.7 ± 0.05 μM, 0.6 ± 0.05 μM, and 0.8 ± 0.15 μM, respectively). Further, 19 exhibited activity against the CCRF‐CEM cell line (CC₅₀ = 0.4 ± 0.05 μM).     

Cellular and Immunologic Injury with PM-10 Inhalation

Abstract

Airborne particles Jess than 10 μsm (PM-10) in mass median aerodynamic diameter (MMAD) are associated with adverse effects on human health including chronic lung diseases and mortality, but the mechanisms by which these particles might cause or aggravate diseases are not specifically known. PM-10 represents a complex mixture, both in terms of size and chemical composition, and it contains both aqueous-media soluble and insoluble particles. Furthermore, the ambient aerosol composition varies markedly in different locations and at different times in the same location. To test the effects of PM-10 on pulmonary defenses in relation to specific cell targets, barrier-reared Sprague-Dawley rats were exposed to purified air (control), to two important constituents of the fine-particle < 1 μm MMAD) fraction of PM-10–ammonium sulfate [(NH₄)₂SO₄^2-] (20 or 70 μg SO₄^2- m^?3, 0.2 μm MMAD) and ammonium nitrate [NH₄NO₃1 (90 or 350 NO₃ μg m ^?3, 0.6 μm MMAD). Rats were also exposed to resuspended road dust (300 and 900 μg m^?3, 4.0 μm MMAD), an important contributor to the coarse (> 2.5 μm MMAD) fraction of PM-10. Exposures were 4 h/day, 4 dayslwk for 8 wk. Macrophage-dependent lung defense functions (antigen binding to Fc receptors and respiratory burst activity) were significantly depressed by NOf, SO₄^2-, and the high-concentration road dust exposures, compared to purified air controls. Lung permeability, as determined from measurements of albumin concentrations in bronchoalveolar lavage fluid, was significantly greater in rats exposed to high concentrations of road dust and NO₃^?, but not to SO₄^2-, when compared to air-exposed controls. Quantitative histopathologic analyses, which included measurements of alveolar nuclear density, alveolar chord length, alveolar septal thickness, and alveolar cross sectional area, showed moderate to substantial changes. In general, the severity of the responses was in the order of SO₄^2-NO₃^?road dust. The findings are consistent with those of epidemiological studies. This study also supports the hypothesis that the fine fraction of PM-10 is more toxic than the coarse fraction.     

新抗肿瘤药铂(Ⅱ),(Ⅳ)配合物的合成

The synthesis of sixteen new analogs of cis-platinum (CDDP) and seven known Pt analogs, as well as the antitumor activity of seven Pt analogs against solid sarcoma 180 are reported. The new congeners consist of Pt (Ⅱ) and Pt (Ⅳ) complexes containing primary amine ligands (eg· NH3, en, pren) with variable anionic ligands such as malonate and 2-substituted malonates. In the experimental antitumor activity tests, 7 new platinum complexes were given on days 1～7, 24 hours after implanted with 10⁶ cells of solid sarcoma 180 tumor and producedT/C (treated/control) value between 15～35% with less toxicity than the parent cisplatinum. The experimental data indicate that it is favorable to introduce certain hydrophilic groups into the platinum complexes, in the view of increasing their aqueous solubility and lowering their toxicity while retaining their effectiveness.     

In vitro pharmacological study of monomeric platinum(III) hematoporphyrin IX complexes

Three stable mononuclear hematoporphyrin IX ((7,12-bis(1-hydroxyethyl)-3,8,13,17-tetramethyl-21H-23H-porphyn-2,18-dipropionic acid), Hp) complexes of Pt^III, namely cis-[ Pt^III(NH₃)₂(Hp_−3H)(H₂O)₂].H₂O 1, [Pt^III(Hp_−3H)(H₂O)₂].H₂O 2 and [Pt^III((O,O)Hp_−2H)Cl(H₂O)₃] 3 with distorted octahedral structure and (d_z2)¹ ground state have been tested in vitro for antineoplastic activity in a panel of tumor cell lines. The novel platinum(III) complexes showed cytotoxic activity in a concentration-dependent manner with IC₅₀ values comparable to those of referent cytotoxic agent cisplatin together with lower cytotoxicity against renal cells. Further detailed evaluation of the active analogue 2 and the less active complex 3 showed that their potency greatly correlates with the ability to induce apoptosis and to bind DNA. Despite the structural dissimilarities between complex 2 and cisplatin, their DNA-adducts were equally effectively recognized and repaired by the nucleotide excision repair system. Complex 2 showed quite superior ability to accumulate in K-562 cells relative to cisplatin.