肝靶向抗病毒药NGA－ACV的制备及其趋肝性

以无唾液酸糖蛋白受体（ａｓｉａｌｏｇｌｙｃｏｐｒｏｔｅｉｎｒｅｃｅｐｔｏｒ，ＡＳＧＰＲ）的特异性配体半乳糖基拟糖白蛋白（ｎｅｏｇｌｙｃｏａｌｂｕｍｉｎ，ＮＧＡ）为载体，通过丁二酰基桥将抗病毒药无环鸟苷（ａｃｙｃｌｏｖｉｒ，ＡＣＶ）与ＮＧＡ偶联，得到肝靶向抗病毒药ＮＧＡＡＣＶ。差热分析和高效液相色谱分析结果表明，ＮＧＡＡＣＶ是共价键偶联物，且在血液中稳定性很好。将偶联物用１３１Ｉ标记后进行家兔放射性显像比较研究。结果，高、低药密度ＮＧＡＡＣＶ的肝脏放射性分别是全身放射性的８１．６％和８６．６％，其趋肝性无明显差别。研究小鼠体内高药密度１３１ＩＮＧＡＡＣＶ的分布，在５ｍｉｎ时肝脏放射性达到峰值，为注入量的８１．７±１０．４％。受体竞争抑制实验表明ＮＧＡＡＣＶ的肝靶向机理为受体介导的主动靶向过程。初步体外抗乙肝病毒比较研究表明，ＮＧＡＡＣＶ较ＡＣＶ的抗病毒剂量有明显降低。     

米托蒽醌聚乳酸缓释毫微粒冻干针剂在动物体内的靶向性研究

目的：比较肝靶向米托蒽醌聚乳酸缓释毫微粒（ＤＨＡＱ－ＰＬＡ－ＮＰ）冻干针剂和ＤＨＡＱ水针剂在小鼠体内的分布规律,验证前者的肝靶向性。方法：采用ＨＰＬＣ法测定静注ＤＨＡＱ－ＰＬＡ－ＮＰ和ＤＨＡＱ水针剂后小鼠血液、心、肝、脾、肺、肾的药物浓度,由此计算各器官的相对百分含量。结果：ＤＨＡＱ－ＰＬＡ－ＮＰ冻干针剂在肝脏的分布明显高于ＤＨＡＱ水针剂,在其它器官中的含量则低于水针剂,给药２４小时后药物在肝中的     

米托蒽醌白蛋白微球和联糖米托蒽醌白蛋白微球肝靶向性比较研究

目的：比较米托蒽醌白蛋白微球和联糖米托蒽醌白蛋白微球肝靶向性的优劣。方法：以小鼠为实验动物,以静脉注射给药后小鼠血浆及各器官中的米托蒽醌含量为指标。结果：联糖米托蒽醌白蛋白微球在肝中的分布高于米托蒽醌白蛋白微球,而且肝中米托蒽醌较高浓度的维持时间也更长。结论：联糖米托蒽醌白蛋白微球肝靶向性比米托蒽醌白蛋白微球好,但无统计学上的显著性差异。     

1-(2,6-二甲基苯氧基)-2-(3,4-二甲氧基苯乙胺基)丙烷盐酸盐对自发性高血压大鼠血管平滑肌细胞增殖及对PDGF-B,bFGF,c-sis,c-myc的影响

用氚胸腺嘧啶核苷（３ＨＴｄＲ）掺入法，电镜，免疫组化，原位杂交方法，在自发性高血压大鼠（ＳＨＲ）观察了１（２，６二甲基苯氧基）２（３，４二甲氧基苯乙胺基）丙烷盐酸盐（ＤＤＰＨ）对血管平滑肌细胞（ＶＳＭＣ）增殖的作用及对生长因子ＰＤＧＦＢ，ｂＦＧＦ及其相关癌基因ｃｓｉｓ与ｃｍｙｃ表达的影响。结果发现：ＤＤＰＨ在降低ＳＨＲ血压同时，能减少肾动脉ＶＳＭＣ的线粒体，粗面内质网和３ＨＴｄＲ掺入量，并能逆转ＶＳＭＣ增殖时ＰＤＧＦＢ，ｂＦＧＦ抗原及ｃｓｉｓ与ｃｍｙｃｍＲＮＡ的表达增强。提示：ＤＤＰＨ能抑制ＳＨＲ的ＶＳＭＣ增殖，与生长因子及癌基因调控的分子生物学机制有关。     

三种阿片受体激动剂对突触传递的抑制作用

目的：比较不同浓度三种阿片受体激动剂对突触传递的抑制作用．方法：用细胞内记录和细胞外微电泳技术，于大鼠伏核脑片制备上记录神经元的兴奋性突触后电位和谷氨酸钠所致细胞膜去极化．结果：（ＤＡｌａ２，ＮＭｅＰｈｅ４，Ｇｌｙｏｌ）ｅｎｋｅｐｈａｌｉｎ（ＤＡＧＯ），（ＤＰｅｎ２，５）ｅｎｋｅｐｈａｌｉｎ（ＤＰＥＮ），ａｎｄｔｒａｎｓ（±）３，４ｄｉｃｈｌｏｒｏＮｍｅｔｈｙｌＮ［２（１ｐｙｒｏｌｉｄｉｎｙｌ）ｃｙｃｌｏｈｅｘｙｌ］ｂｅｎｚｅｎｅａｃｅｔａｍｉｄｅｍｅｔｈａｎｅｓｕｌｆｏｎａｔｅ（Ｕ５０４８８Ｈ）（１μｍｏｌ·Ｌ－１）均降低突触传递，尤以ＤＡＧＯ抑制作用最著．结论：三种阿片受体激动剂对突触传递的抑制作用均具有浓度依赖性，其作用机制与谷氨酸介导的突触传递受抑制有关．     

米托蒽醌毫微球在裸小鼠人肝癌模型体内的靶向性研究

采用液体闪烁计数技术研究厂氖标记米托惠醌聚氰基丙烯酸正丁酯毫微球（￣（３）Ｈ－ＤＨＡＱ－ＰＢＣＡ－ＮＳ）在常位及异位（腋下）裸小鼠人肝癌模型体内各脏器、肌肉、肝常位肿瘤组织和腋下肿瘤组织中的分布。证明该制剂具有良好的肝靶向作用，肝脏中的含量为体内总药量的７１．３１±１０．４９％（ｎ＝５）。并发现其在常位肿瘤组织中的含量高于肝组织中的含量，腋下肿瘤组织中的含量高于腋下肌肉组织中的含量，但均无显著性差异。为该制剂的进一步研究提供了一定的科学依据。     

肝靶向抗疟药半乳糖基拟糖白蛋白－伯氨喹偶联物和磷酸伯氨喹的药代动力学

用ＨＰＬＣ法对肝靶向抗疟药ＮＧＡ－ＰＱ各磷酸伯氨喹（ＰＱＰ）在小鼠体内的药代动力学行为进行了比较研究。结果表明ＮＧＡ－ｐＱ在血中有较好的稳定性，不易解离出ＰＱ。ＮＧＡ－ＰＱ和ＰＱＰ在肝中的Ｔｍ分别为１０和１５ｍｉｎ，在血中的Ｔ１／２分别为２０．４４和３５．７４ｍｉｎ。在肝中的Ｔ１／２分别为４３．９５和２１．４６ｍｉｎ，肝中的ＡＵＣ分别为２３０５．８０和３３３．２９ｍｉｎ·μｇ－１·ｇ－１。说明ＮＧＡ－ＰＱ在血中很快消除并浓集于肝脏，在肝脏的保留时间长，从而证实ＮＧＡ－ＰＱ具肝靶向分布特性。     

Antisense oligodeoxynucleotides downregulated c sis mRNA expression to inhibit proliferation of vascular smooth muscle cells 1

目的：研究ｃｓｉｓ反义寡脱氧核苷酸（ＯＤＮ）对ｃｓｉｓ表达及血管平滑肌细胞（ＶＳＭＣ）增殖抑制效应．方法：用合成ｃｓｉｓ正、反义ＯＤＮ培养ＶＳＭＣ;用液闪测定［３Ｈ］ＴｄＲ掺入并细胞计数,观察细胞增殖;用逆转录ＰＣＲ,评价ｃｓｉｓ表达．结果：ｃｓｉｓ反义ＯＤＮ２,４,６,８,１０μｍｏｌ·Ｌ－１抑制ＶＳＭＣ（１０３％±０７％,２２６％±０９％,３１０％±１１％,３５４％±０９％,４３３％±１２％）和降低［３Ｈ］ＴｄＲ掺入（６８％±０３％,９７％±０７％,２９０％±０６％,３２０％±０７％,５０６％±１３％）呈有剂量依赖性．反义ＯＤＮ１０μｍｏｌ·Ｌ－１培养细胞４ｄ,最大抑制率达６０３％±１０％,［３Ｈ］ＴｄＲ掺入降低５６３％±０９％,ｃｓｉｓｍＲＮＡ表达明显降低;而正义ｃｓｉｓＯＤＮ对ＶＳＭＣ无抑制,细胞数和［３Ｈ］ＴｄＲ掺入及ｃｓｉｓｍＲＮＡ水平与对照无差异．结论：ｃｓｉｓ反义ＯＤＮ明显下调ｃｓｉｓｍＲＮＡ表达,显著抑制ＶＳＭＣ增殖     

反相高效液相色谱法测定慢性肾功能衰竭家兔肾组织中的磷酸腺苷

研究测定慢性肾功能衰竭家兔肾组织中ＡＴＰ,ＡＤＰ,ＡＭＰ的含量。方法：用高效液相色谱法（ＨＰＬＣ）,ＵＬＴＲＡＳＰＨＥＲＥ（ＴＭ）Ｃ１８柱（４．６ｍｍ×２５ｃｍ）,流动相为甲醇磷酸缓冲液（ｐＨ５．９,３０ｍｍｏｌ·Ｌ－１）（３∶９７）,检测波长２５４ｎｍ,在６ｍｉｎ内检测完毕。结果：家兔肾组织中磷酸腺苷ＡＴＰ,ＡＤＰ,ＡＭＰ）的含量分别为８９．６±１０．２,１１２．４±８．９,７８．４±６．２μｇ·ｇ－１（湿组织）,平均回收率分别为１００．９％,９９．９％,１０１．０％。结论：此方法简便,快速,准确,可用于药物监测及基础研究。     

甘糖酯对家兔全脑缺血再灌损伤的保护作用

甘糖酯是一种新型的低分子量、低抗凝活性的多糖类海洋药物。本文报道甘糖酯对４动脉阻断家兔全脑缺血再灌损伤脑组织过氧化脂质（ＭＤＡ）、超氧化物歧化酶（ＳＯＤ）、谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）、一氧化氮（ＮＯ）和脑含水量的影响。结果显示，甘糖酯可降低全脑缺血再灌损伤的ＭＤＡ含量，提高ＳＯＤ和ＧＳＨ－Ｐｘ活性，降低脑含水量，对ＮＯ无明显影响，提示甘糖酯对全脑再灌损伤有明显的抗氧化作用。     

肝靶向药物研究——模型化合物NGA-去甲替林的合成及其趋肝性

肝细胞膜上的肝结合蛋白(HBP)是无唾液酸糖蛋白的受体。作者合成了无唾液酸糖蛋白的类似物—半乳糖新糖白蛋白(NGA)并用其作为肝靶向药物的载体,采用琥珀酸酐法与药物去甲替林(NT)偶联得NGA-NT。以此作模型化合物,经~(99m)Tc标记后的动物放射显像及HPLC检测结果,说明NGA-NT在血中稳定,在肝脏降解释出NT,具有显著的趋肝性。     

肝靶向抗癌药NGA-DHAQ的合成研究

以D－半乳糖和人血清白蛋白（HSA）合成无唾液酸糖蛋白受体的特异性配体一半乳糖基拟糖白蛋白（neoglycoalbumin,NGA）;以NGA为载体通过丁二酰基桥将米托蒽醌（mitoxantrone,DHAQ）与其偶联,得到肝靶向药物NGA－DHAQ,紫外光谱和HPLC分析表明：NGA－DHAQ为化学偶联物,在血液中稳定。     

99mTc-neoglycoalbumin (NGA)-binding to human hepatic binding protein (HBP) in vitro.

1 Neoglycoalbumin (NGA) was synthesised by covalent coupling of 2-imino-2-methoxyethyl-1-thio-beta-D-galactopyranoside (IME-thiogalactose) to the primary amino groups of human serum albumin (HSA). NGA was purified by ultrafiltration and size exclusion h.p.l.c. (SEC). 99mTc-labelling was performed with and without SEC purification. 2 Estimation of 99mTc-NGA-binding to human hepatic binding protein (HBP) revealed a complex behaviour indicating saturable high- and low-affinity sites. The high-affinity binding capacity was 1.1 +/- 0.4 pmol mg-1 human liver plasma membrane protein, the low-affinity binding capacity was 6.2 +/- 1.8 pmol mg-1 liver plasma membrane protein. The apparent equilibrium dissociation constants were 2.4 +/- 1.2 and 18.4 +/- 4.8 nM, respectively. 3 Specific binding of 99mTc-NGA to human HBP in the presence of 100 microM unlabelled NGA, Ca++ and Mg++ at pH 7.5 and 37 degrees C reached 85 +/- 5% at equilibrium. The amount of ligand specifically bound increased with the amount of human liver membrane protein added. The concentration of unlabelled agonist necessary to displace 50% of ligand bound amounted to 100 nM.     

肝靶向抗病毒药NGA-ACV的制备及其趋肝性

以无唾液酸糖蛋白受体(asialoglycoprotein receptor,ASGP-R)的特异性配体——半乳糖基拟糖白蛋白(neoglycoalbumin,NGA)为载体,通过丁二酰基桥将抗病毒药无环鸟苷(acyclovir,ACV)与NGA偶联,得到肝靶向抗病毒药NGA-ACV。差热分析和高效液相色谱分析结果表明,NGA-ACV是共价键偶联物,且在血液中稳定性很好。将偶联物用¹³¹I标记后进行家兔放射性显像比较研究。结果,高、低药密度NGA-ACV的肝脏放射性分别是全身放射性的81.6%和86.6%,其趋肝性无明显差别。研究小鼠体内高药密度¹³¹I-NGA-ACV的分布,在5min时肝脏放射性达到峰值,为注入量的81.7±10.4%。受体竞争抑制实验表明NGA-ACV的肝靶向机理为受体介导的主动靶向过程。初步体外抗乙肝病毒比较研究表明,NGA-ACV较ACV的抗病毒剂量有明显降低。     

Design and synthesis of novel galactosylated polymers for liposomes as gene drug carriers targeting the hepatic asialoglycoprotein receptor

The 18-mer oligodeoxynucleotides (ODNs) that can inhibit survivin gene expression were selected as a model gene drug to study hepatic-targeting drug delivery system. Novel galactosylated polymers (cholesteryloxycarbonylamino) ethylamine-α,β-polyasparthydrazied (CHE-PAHy-Lacs), which target asialoglycoprotein receptor on hepatic parenchymal cells (PC), were designed and synthesized as non-toxic, non-antigenic and non-teratogenic ligands for liposomes. The liposomes incorporating different CHE-PAHy-Lacs were prepared and characterized by zeta potential and particle size analyzer. The drug encapsulation efficiency was measured by gel filtration method. 1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate was used as a marker for all the liposome preparations in the in vivo experiments. The CHE-PAHy-Lac liposomes produced a significant improvement in the encapsulation efficiency of ODNs (28.73–51.37%) compared with conventional liposomes (9.88%). The in vivo results showed that the liposomes incorporating CHE-PAHy-Lac, which contained about 30% (w/w) galactosyl residues, exhibited marked accumulation in the liver and hepatic PC. These results suggest that the novel galactosylated polymers used for liposomes have a great potential as a gene delivery system for hepatic targeting.     

肝动脉栓塞米托蒽醌乙基纤维素微球的研究

利用正交实验设计法,优选适用于肝动脉栓塞的米托蒽醌乙基纤维素微球制备条件和工艺;采用动态透析法研究了该微球的体外释药规律;根据混悬液的稳定性理论,优选并制备了适于临床肝动脉介导栓塞使用的米托蒽醌乙基纤维素微球混悬注射液。结果表明∶在优化工艺条件下制得的米托蒽醌乙基纤维素微球外形圆整,球径在40～200μm范围内的占总数的91.9%,平均球径为110.24±38.19μm;包封率为55.6%;载药量为12.5%;体外释药符合单指数模型,释药方程为lg(Y_∞-Y)=-0.116t-1.198×10^-3(γ=0.9992,t₅₀=2.6h);其混悬液适于临床应用。用狗进行的实验表明肝血药浓度高,平均驻留时间比注射剂长2.45倍。     

肺靶向米托蒽醌明胶微球的研究

采用二步法制备米托蒽醌明胶微球,球径范围为5.1～25.0μm的占总数87.36%,体外释药与原药相比t1/2延长4倍,DTA曲线上的特征吸热峰为133℃,经37℃,RH75%考察3月,几乎无变化。经小鼠体内分布试验表明具有明显的肺靶向性,靶向效率增加3～35倍,肺中药代动力学行为可用一室开放模型描述,平均滞留时间延长10h。     

反义核酸药物脂质体肝靶向性的研究

目的研究反义核酸药物脂质体给药系统的肝主动靶向性。方法采用薄膜分散法制备脂质体,以花青染料荧光标记脂质体,以荧光分光光度法定量分析;利用肝组织的体内冷冻切片和小鼠体内组织分布研究肝靶向性;检测大鼠肝非实质细胞与肝实质细胞的荧光强度(INPC和IPC)。结果主动靶向脂质体在肝脏中的荧光强度明显强于非主动靶向脂质体,相对摄取率(re)为5.64;非主动靶向脂质体的荧光值比(IPC/INPC)为1.16±0.89,主动靶向脂质体的IPC/INPC为8.24±1.37。结论主动靶向脂质体具有良好的肝实质细胞靶向性和肝靶向性。     

Trans-nasal zolmitriptan novasomes: in-vitro preparation,optimization and in-vivo evaluation of brain targeting efficiency

Migraine attack is a troublesome physiological condition associated with throbbing, intense headache, in one half of the head. Zolmitriptan is a potent second-generation triptan, prescribed for patients with migraine attacks, with or without an aura, and cluster headaches. The absolute bioavailability of zolmitriptan is about 40% for oral administration; due to hepatic first metabolism. Nasal administration would circumvent the pre-systemic metabolism thus increasing the bioavailability of zolmitriptan. In addition, due to the presence of microvilli and high vasculature, the absorption is expected to be faster compared to oral route. However, the bioavailability of nasal administered drugs is particularly restricted by poor membrane penetration. Thus, the aim of this work is to explore the potential of novel nanovesicular fatty acid enriched structures (novasomes) for effective and enhanced nasal delivery of zolmitriptan and investigate their nose to brain targeting potential. Novasomes were prepared using nonionic surfactant, cholesterol in addition to a free fatty acid. A 2³ full factorial design was adopted to study the influence of the type of surfactant, type of free fatty acid and ratio between the free fatty acid and the surfactant on novasomes properties. The particle size, entrapment efficiency, polydispersity index, zeta potential and % zolmitriptan released after 2?h were selected as dependent variables. Novasomes were further optimized using Design Expert® software (version 7; Stat-Ease Inc., Minneapolis, MN), and an optimized formulation composed of Span® 80:Cholesterol:stearic acid (in the ratio 1:1:1) was selected. This formulation showed zolmitriptan entrapment of 92.94%, particle size of 149.9?nm, zeta potential of ?55.57?mV, and released 48.43% zolmitriptan after 2?h. The optimized formulation was further examined using transmission electron microscope, which revealed non-aggregating multi-lamellar nanovesicles with narrow size distribution. DSC, XRD examination of the optimized formulation confirmed that the drug have been homogeneously dispersed throughout the novasomes in an amorphous state. In-vivo bio-distribution studies of ^99mTc radio-labeled intranasal zolmitriptan loaded novasomes were done on mice, the pharmacokinetic parameters were compared with those following administration of intravenous ^99mTc-zolmitriptan solution. Results revealed the great enhancement in zolmitriptan targeting to the brain, with drug targeting potential of about 99% following intranasal administration of novasomes compared with the intravenous drug solution. Zolmitriptan loaded novasomes administered via the nasal route may therefore constitute an advance in the management of acute migraine attacks.