注射用胸腺肽辅料的研究

目的：寻找一种注射用胸腺肽适用的辅料。方法：以三批同一批次的胸腺肽原料液为实验对象,通过Folin-酚法和T细胞活性测定法-脱E受体法研究了三种辅料对胸腺肽多肽含量及活性的影响。结果：7％甘露醇作为辅料时使胸腺肽多肽含量测定显著增高,而右旋糖酐则不影响测定结果,三种辅料对胸腺肽活性均无显著影响。结论：最终选用4％右旋糖酐作为注射用胸腺肽的辅料。     

Antagonistic effect of nerve growth factor on neuronal injury induced by hypoxia in cultured cerebral cortical neurons of rats

目的：以连二亚硫酸钠造成原代培养的大鼠胎鼠脑皮质神经元缺氧损伤,观察神经生长因子（ＮＧＦ）对缺氧损伤神经元的影响．方法：测定神经元生存力及细胞外液乳酸脱氢酶（ＬＤＨ）的活性来分析ＮＧＦ的作用,脑皮质细胞匀浆用于测定丙二醛（ＭＤＡ）含量及超氧化物歧化酶（ＳＯＤ）和谷胱甘肽过氧化物酶（ＧＳＨＰｘ）活性．结果：缺氧后,ＬＤＨ释放及细胞生存力降低．ＮＧＦ（１－１００μｇ·Ｌ－１）浓度依赖地减少ＬＤＨ的释放及ＭＤＡ的生成,ＮＧＦ１００μｇ·Ｌ－１显著提高细胞生存力及提高ＳＯＤ和ＧＳＨＰｘ活性２７倍．结论：ＮＧＦ通过减少脂质过氧化物生成及提高ＳＯＤ和ＧＳＨＰｘ活性来保护大脑皮质细胞抗缺氧损伤     

赋形剂对注射用促肝细胞生长素含量测定的影响

目的 :研究赋形剂对注射用促肝细胞生长素含量测定的影响。方法 :根据注射用促肝细胞生长素的处方[1 ] ,考察甘露醇及右旋糖酐对多肽含量测定影响的程度。结果 :实验表明甘露醇在 0 32～ 1 6mg ml浓度范围内 ,与吸收度呈线性关系 ;右旋糖酐与吸收度呈相关性 ,但不呈线性关系 ;原料溶液按处方量加入甘露醇和右旋糖酐[1 ] ,含量平均增加 4 82μg ml。结论 :甘露醇在赋形剂中所占比例较高 ,对含量测定影响较大 ,而右旋糖酐在赋形剂中所占比例较少 ,不影响含量测定     

人卵促性腺激素释放肽（hF－GRP）及其类似物合成和生物活性初探

用固相法合成了ｈＦ－ＧＲＰ及其１５个类似物。全部裂解均用三氟甲磺酸完成。产物总收率６０％～８０％。对所有合成肽进行了影响离体的小鼠垂体分泌ＬＨ的活性筛选。结果表明，当合成肽的浓度为０．０５ｍｍｏｌ／Ｌ时：（１）将ｈＦ－ＧＲＰ的Ｃ端ＣＯＯＨ变成ＣＯＮＨ２，活性变化不大；（２）Ｃ端残基Ａｓｎ１４被Ｐｈｅ替换后刺激垂体分泌ＬＨ的活性明显高于ｈＦ－ＧＲＰ；（３）Ｔｈｒ３被Ｔｙｒ替换后片段ｈＦ－ＧＲＰ（３～１３）有抑制ＬＨ分泌的活性；（４）其余类似物与空白对照相似。     

药物对映体含量测定的核磁共振法研究──Ⅱ．3，4－环氧－3，4－二氢－2，2－二甲基－6－乙酰胺基－7－硝基－二氢－1－苯骈吡喃

报道了位移试剂Ｅｕ（ＦＯＤ）３、Ｅｕ（ＨＦＣ）３，存在下３，４－环氧－３，４－二氢－２，２－二甲基－６－乙酰胺基－７－硝基－二氢－１－苯骈吡喃（Ⅰ）的镧诱导位移（ＬＩＳ）值；在手性位移试剂Ｅｕ（ＨＦＣ）３存在下，观察到５－Ｈ和８－Ｈ明显的对映体位移差值（△△δ），据此，可对化合物Ⅰ直接进行对映体的定量分析。当Ｅｕ（ＨＦＣ）３：Ⅰ为０．９５５（０．７０５）（摩尔比）时，８－Ｈ（５－Ｈ）的（＋）和（－）对映体两峰峰谷高度为峰高的３．５％（７．１％），即当对映体之一的含量不低于３．５％（７．１％）时，可对化合物Ⅰ的对映体含量进行精确的直接测定。     

4-〔4'-(2',2',6',6'-四甲基哌啶氮氧自由基)氨基〕-4'-去甲表鬼臼毒及其自由基还原物抗肿瘤作用比较

目的：比较鬼臼毒氮氧自由基衍生物４－〔４″－（２″，２″，６″，６″－四甲基哌啶氮氧自由基）氨基〕－４′－去甲表鬼臼毒（ＧＰ－７）及其自由基还原物ＧＰ－７－Ｈ和ＧＰ－７－ＯＨ的抗肿瘤活性和急性毒性。方法：采用小鼠移植性肿瘤Ｓ１８０、ＨｅＰＳ和腹腔注射ＬＤ５０值。结果：ＧＰ－７、ＧＰ－７－Ｈ、ＧＰ－７－ＯＨ５～１０ｍｇ·ｋｇ－１给药１０ｄ，对Ｓ１８０肉瘤的抑制率分别为３９．７％～４６．８％、１７．３％～２９．５％和１９．９％～２２．４％，对ＨｅＰＳ的抑制率分别为３８．７％～４８．８％、１５．５％～３５．１％和１８．４％～３３．３％。昆明种小鼠腹腔注射一次给药，ＬＤ５０分别为２３１．２、８９．７和１２９．５ｍｇ·ｋｇ－１。结论：ＧＰ－７对Ｓ１８０、ＨｅＰＳ生长抑制作用均较其还原物ＧＰ－７－Ｈ、ＧＰ－７－ＯＨ强，急性毒性较其还原物ＧＰ－７－Ｈ和ＧＰ－７－ＯＨ小；提示ＧＰ－７中的自由基在加强抗肿瘤作用和降低毒性方面起着重要的作用。     

Effect of artemether on glyceraldehyde-3-phosphate dehydrogenase,phosphogly-cerate kinase,and pyruvate kinase of Schistosoma japonicum harbored in mice

目的：研究蒿甲醚（Ａｒｔ）对日本血吸虫３磷酸甘油醛脱氢酶（ＧＡＰＤＨ）、磷酸甘油酸激酶（ＰＧＫ）和丙酮酸激酶（ＰＫ）的影响．方法：小鼠感染血吸虫尾蚴３２－３８ｄ后ｉｇＡｒｔ１００－３００ｍｇ·ｋｇ－１，２４－７２ｈ后取虫测定上述３种酶和乳酸含量．结果：小鼠ｉｇＡｒｔ３００ｍｇ·ｋｇ－１后２４－４８ｈ，血吸虫♀、♂虫的ＰＧＫ和ＰＫ活力被抑制２７％－４８％；♀虫的ＧＡＰＤＨ对Ａｒｔ亦较敏感，♂虫则否．给药后７２ｈ，♀虫乳酸含量降低７２％，♂虫的降低４９％．结论：Ａｒｔ对血吸虫的ＰＧＫ和ＰＫ活力有明显抑制作用，♀虫的ＧＡＰＤＨ对Ａｒｔ亦较敏感，♀虫乳酸含量降低较♂虫明显     

Effect of basic fibroblast growth factor on focal ischemic injury and antioxidant enzyme activities

目的：考察碱性成纤维细胞生长因子（ｂＦＧＦ）对局部脑缺血损伤和抗氧酶活性的影响．方法：大鼠用插丝法阻塞大脑中动脉２４ｈ,梗塞灶用ＴＴＣ染色显示,图象分析测量;神经功能缺损采用０－５级评分．分别分析每只大鼠左右两侧大脑的４种抗氧化酶活性水平．结果：ｂＦＧＦ（缺血５ｍｉｎ后以４５μｇ·ｋｇ－１·ｈ－１给予３ｈ）可降低梗塞灶体积和神经功能缺损评分．局部脑缺血升高过氧化氢酶（ＣＡＴ）,超氧化物歧化酶（ＳＯＤ）和谷胱甘肽过氧化物酶（ＧＳＨＰＸ）的活性,但使谷胱甘肽Ｓ转移酶（ＧＳＨＳＴ）的活性水平降低．ｂＦＧＦ可在缺血状态下进一步升高ＣＡＴ的活性,但对ＳＯＤ,ＧＳＨＰＸ和ＧＳＨＳＴ的水平没有影响．结论：ｂＦＧＦ对局部脑缺血损伤具有神经保护作用,此作用可能与ＣＡＴ活性水平的升高有关．     

人白细胞干扰素和基因工程干扰素α-1b治疗慢性乙型肝炎

目的：比较人白细胞干扰素（ＬＩＦＮ）和基因工程干扰素α－１ｂ（ｒＩＦＮα－１ｂ）治疗慢性乙型肝炎（ＣＨＢ）的疗效和不良反应。方法：ＣＨＢ病人８３例（男性６９例，女性１４例；年龄３２±ｓ８ａ）。Ａ组２９例，用ＬＩＦＮ３ＭＵ；Ｂ组２７例，用ｒＩＦＮα－１ｂ３ＭＵ，Ｃ组２７例用人血浆白蛋白冻干制品１０ｇ，３组均ｉｍ，ｑｄ×４ｗｋ，继以ｑｏｄ×８ｗｋ。结果：３组ＨＢｅＡｇ阴转率分别为３８％，４１％，７％；ＨＢＶ－ＤＮＡ阴转率分别为４０％，５２％，７％。Ａ和Ｂ组ＩＦＮ对ＨＢＶ复制的抑制作用显著优于Ｃ组（Ｐ＜０．０５），但Ａ和Ｂ组间无显著差异。Ａ组不良反应显著大于Ｂ组（Ｐ＜０．０５）。结论：２种不同来源的ＩＦＮ治疗ＣＨＢ均有显著疗效，２药无显著差别，但Ａ组不良反应明显。     

神经生长因子体外生物学活性测定国际标准方法的初步建立

建立神经生长因子（ＮＧＦ）体外生物学活性测定的国际标准方法。方法：利用ＭＴＴ法测定ＮＧＦ促进ＴＦ－１细胞的增殖活性。结果：５～１０ ｎｇ／ｍｌ ＮＧＦ能明显促进ＴＦ－１细胞的增殖。结论：初步建立的 ＮＧＦ体外生物学活性测定的国际标准方法稳定可靠，简便易行。     

甘露醇对福林-酚法测定注射用P-转移因子中多肽含量的影响

采用甘露醇和牛血清白蛋白为对照,消除了甘露醇对注射用P-转移因子中多肽测定的影响.线性浓度范围;甘露醇0～1.3mg/ml、牛血清白蛋白4.3～10.8μg/ml;平均回收率为102.l%相对标准偏差为0.85%(n=4).     

Lyoprotection of aviscumine with low molecular weight dextrans

The aim of this research was to ascertain whether dextrans with low molecular weight will stabilize aviscumine. During freeze-drying increasing concentrations of dextran T1 (MW 1000) stabilized aviscumine. Eight percent of dextran resulted in a nearly 100% recovery of the activity and in addition a complete amorphous structure of the solid phase was obtained. By decreasing the molecular weight of the dextran from 75 to 1 kDa, the protein activity was increased by 20% in the lyophilisate. Combinations of dextran with either trehalose or mannitol showed no additional effects on stability. The improved stabilization of aviscumine using low molecular weight dextrans is explained by an increased interaction between the protein and the dextran molecules (like hydrogen bonds), whereas they are sterically hindered if larger dextran molecules are used. When the protein concentration was increased from 10 to 100 microg/ml (in formulas with 8% dextran T1), no influence on the protein activity could be found. With regard to the carbohydrate-binding activity of the protein, it was shown that the optimal content of residual water in the lyophilisate should be about 2%. Above and below this percentage a destabilization of the protein was observed. The often discussed failure of dextran as a stabilizing excipient in the freeze-drying of proteins seems to be a question of the selection of the correct molecular weight.     

注射用促肝细胞生长素的质量考察

目的研究注射用促肝细胞生长素的质量。方法测定水分、游离氨基酸、稳定剂、赋形剂含量 ,并考察制剂外观。结果与正常组比较 ,萎缩组游离氨基酸含量显著增加 (P <0 .0 1 ) ,褐变组稳定剂含量显著减少 (P <0 .0 1 ) ,水分含量增加 (P <0 .0 5)。结论在现有工艺条件下 ,注射用促肝细胞生长素外观质量与游离氨基酸、稳定剂、水分有关 ,控制其含量有助保证质量     

Physical characteristics and aerosolization performance of insulin dry powders for inhalation prepared by a spray drying method

The objective of this study was to investigate the influence of formulation excipients on the physical characteristics and aerosolization performance of insulin dry powders for inhalation. Insulin dry powders were prepared by a spray drying technique using excipients such as sugars (trehalose, lactose and dextran), mannitol and amino acids (L-leucine, glycine and threonine). High performance liquid chromatography and the mouse blood glucose method were used for determination of the insulin content. The powder properties were determined and compared by scanning electron microscopy, thermo-gravimetric analysis and size distribution analysis by a time-of-flight technique. The in-vitro aerosolization behaviour of the powders was assessed with an Aerolizer inhaler using a twin-stage impinger. Powder yield and moisture absorption were also determined. Results showed that there was no noticeable change in insulin content in any of the formulations by both assay methods. All powders were highly wrinkled, with median aerodynamic diameters of 2-4 microm, and consequently suitable for pulmonary administration. The tapped density was reduced dramatically when glycine was added. The powders containing mannitol, with or without L-leucine, were less sensitive to moisture. The highest respirable fraction of 67.3 +/- 1.3% was obtained with the formulation containing L-leucine, in contrast to formulations containing glycine and threonine, which had a respirable fraction of 11.2 +/- 3.9% and 23.5 +/- 2.5%, respectively. In addition, powders with good physical properties were achieved by the combination of insulin and trehalose. This study suggests that L-leucine could be used to enhance the aerosolization behaviour of the insulin dry powders for inhalation, and trehalose could potentially be used as an excipient in the formulations.     

Effects of Excipients on Protein Conformation in Lyophilized Solids by Hydrogen/Deuterium Exchange Mass Spectrometry

Purpose Excipients are added to lyophilized protein drug formulations to protect the protein during processing and storage, but the mechanisms are poorly understood. Here, hydrogen/deuterium (H/D) exchange with mass spectrometry was used to assess protein conformation and excipient interactions in lyophilized solids. Methods Calmodulin (CaM, 17 kD) was co-lyophilized with carbohydrate excipients (sucrose, mannitol, trehalose, raffinose, dextran 5,000, dextran 12,000) or guanidine hydrochloride (negative control) and exposed to D₂O vapor at 33% RH and RT. Samples were then dissolved and analyzed by mass spectrometry (+ESI/MS). Peptic digestion provided additional, site-specific information on H/D exchange. Solids were further characterized by powder x-ray diffraction (PXRD), differential scanning calorimetry (DSC), infrared spectroscopy (FTIR) and water vapor sorption. Results Excipients protected CaM from H/D exchange, increasing in the order guanidine hydrochloride < no excipient, mannitol < dextran 5,000, dextran 12,000 < sucrose < raffinose < trehalose. Effects were exerted primarily in the protein’s α-helical segments. Conclusions The effects of carbohydrate excipients on protein conformation in lyophilized solids are not exhibited uniformly along the protein sequence, but instead are exerted in a site-specific manner. The results also demonstrate the utility of H/D exchange with ESI/MS for protein structure characterization in lyophilized samples.     

Estimation of the percolation thresholds in ternary lobenzarit disodium–dextran–HPMC hydrophilic matrices tablets: Effects of initial porosity

The aim of this work is to estimate the excipient percolation threshold for a new combined matrix native dextran (DT), series B110-1-2 (Mw 2 × 10⁶): HPMC K4M CR: lobenzarit disodium (LBD) system and demonstrate the advantages of this ternary system with respect to previously reported binary dextran:LBD and HPMC:LBD tablets. The formulations studied were prepared with different amounts of excipient (DT:HPMC, 4:1 (wt/wt) for all tablets and relative polymer/drug particle size of 4.17) in the range of 10–70% (wt/wt). Dissolution studies were carried out using the paddle method (100 rpm) and one face water uptake measurements were performed using a modified Enslin apparatus. The Higuchi's models as well as the non-linear regression were employed as empiric methods to study the released data. Values of diffusion exponent 0.588 < n < 0.784 (Korsmeyer equation) for dissolution profile and water uptake mechanism 0.715 < n < 0.960 (Davidson and Peppas equation) suggests anomalous or complex mechanisms in all cases. The critical points in ternary tablets were reduced from 44.75% (v/v) of excipient (correspond to purely native dextran) to 22.34% (v/v) (corresponding to mixture native dextran:HPMC, 4:1, wt/wt). The initial porosity (IP) of hydrophilic matrices above the values of 20% has an important influence on the percolation threshold as well as on establishment of the gel barrier responsible for the controlled release from the DT:HPMC:LBD tablets.     

注射用雷莫司琼的制备

目的:探讨注射用雷莫司琼的处方组成、测定方法与制备工艺。方法:以甘露醇、山梨醇、右旋糖酐为填充剂,制备了注射用雷莫司琼;采用HPLC测定注射用雷莫司琼的含量及有关物质。结果:最佳处方为甘露醇100 g,雷莫司琼0．3 mg;3批样品的含量分别为100．5％,101．2％,99．6％,有关物质的含量分别为 0．17％,0．17％,0．18％,符合新药申报要求。结论:处方及工艺经中试放大制备的注射用粉针性质稳定;质量可控。     

Dextran or hydroxyethyl starch in spray-freeze-dried trehalose/mannitol microparticles intended as ballistic particulate carriers for proteins

The goal of this study was to clarify the effects of dextran 10 kDa on the properties of spray-freeze-dried microparticles for use with ballistic injectors. A novel carrier of trehalose, mannitol, and the polymer is known to maximize particle density. Measurements of T'(g) showed that the dextran anti-plasticizes the trehalose/mannitol, but also undergoes phase separation. The product temperature exceeded T'(g) during primary drying. The collapsed particles can therefore be explained by plastic flow of the freeze concentrate. DSC of the powder showed T(g) at 45 degrees C and, in the first scan, a wide endothermic melting peak caused by mannitol recrystallization. Catalase showed 35% activity loss on rehydration of its spray freeze-drying (SFD) powder, which was improved in the TM/D (3:3:4) formulation, but not up to that level seen with either trehalose or mannitol alone. The dextran 10 kDa, which is vital to maximize particle density, was therefore detrimental to protein integrity during SFD, as also found with a 65-72 kDa dextran. Hydroxyethyl starch (HES) 200 kDa gave similar, limited stabilizing effects on the protein. The proportion of polymer in the formulation should be low to minimize protein damage, whilst high enough to give required particle morphology and density.     

Do co-spray dried excipients offer better lysozyme stabilisation than single excipients?

The inherent instability of proteins when isolated from their native conditions creates the necessity of suitable stabilisation techniques. Because of the instability of proteins in solution it is often necessary to produce them as solid formulations. A method of producing relatively stable, solid protein pharmaceuticals is to incorporate them with a suitable excipient into an amorphous matrix by dehydration. The use of spray dried multiple excipient/single protein blends was compared to single excipient/protein systems using lysozyme as a model protein to establish the stabilising ability of such systems. Unprocessed controls and spray dried samples were characterised structurally by X-ray powder diffraction and Fourier transform Raman spectroscopy and also thermally by differential scanning calorimetry and thermogravimetric analysis. Retained lysozyme activity was assayed enzymatically. To assess long-term stability, samples were subjected to conditions of elevated temperature and relative humidity (RH) 40 degrees C/75% RH. Structural and thermal analysis of samples revealed that mannitol/trehalose spray dried excipient/lysozyme blends were completely amorphous upon production but partially recrystallised upon storage at elevated temperature and RH. Biological activity assays revealed that samples containing trehalose retained the highest percentage activity. Under the conditions employed mannitol/trehalose systems stabilise lysozyme more effectively than single excipient systems due to their ability to form amorphous products.