不同配方冻干保护剂对猪源纤维蛋白原冻干质量的影响

目的研究不同配方冻干保护剂与猪源纤维蛋白原冻干质量的关系,为猪源纤维蛋白原制剂的质量控制提供参考。方法采用冷冻干燥法制备猪源纤维蛋白原制剂,并分别加入A(组氨酸)及B、C、D 4种配方保护剂缓冲液(精氨酸浓度:B 相似文献   

注射用5-氮杂胞苷的制备及质量研究

目的 采用冷冻干燥工艺制备注射用5-氮杂胞苷,并对其质量进行评价。方法 通过对5-氮杂胞苷溶液的稳定性研究确定最佳溶液配制温度。根据样品外观确定预冻工艺参数。根据样品温度和干燥时间采用析因实验设计优化一次干燥过程中隔板温度和真空度。根据水分、有关物质结果确定二次干燥的温度及时间。根据优化的冻干工艺参数制备3批注射用5-氮杂胞苷,并与参比制剂进行对比研究。结果 5-氮杂胞苷溶液对温度极其敏感,需在(4±2)℃条件下配制,在6 h内药物较为稳定;采用反复冻结法,降温速率为0.5～1℃·min^-1,退火温度-10℃及保持时间为2 h,冻干得到的注射用5-氮杂胞苷外观均匀、无鳞片状。通过实验优化得到最佳一次干燥工艺参数为：隔板温度为20℃,真空度为60 Pa,在该工艺参数范围内进行操作,冻干时间明显缩短;二次干燥温度为30℃,干燥时间为40 h,产品水分与有关物质均可以达到与参比制剂相似;三批自制注射用5-氮杂胞苷的检测指标与参比制剂相似或一致。结论 本研究作者按照优化的冷冻干燥工艺参数制备注射用5-氮杂胞苷,可以得到与参比制剂质量一致的制剂产品。     

卡巴他赛纳米脂质体冻干工艺的考察

为了提高贮存稳定性、降低乙醇残留量,将卡巴他赛纳米脂质体制备成冻干注射剂,并对冻干工艺进行了研究与优化。以冻干产品的外观、主药含量、包封率、水分含量、乙醇残留量、贮存稳定性为评价指标,考察了冻干保护剂(蔗糖)用量、料液装量、预冻速度、干燥温度对产品质量的影响。结果表明,蔗糖与磷脂的质量比为5.0时较佳;脂质体中卡巴他赛的质量浓度为2 mg/mL;装量为每瓶20 mg;西林瓶选用20或30 mL 2个规格。优化冻干工艺参数为：预冻温度–40℃,隔板降温时间90 min,–40℃时保温5 h;一次干燥温度–15℃,一次干燥时间14 h;二次干燥温度28℃,二次干燥时间28 h;冻干全程总耗时48.83 h;干燥过程采用阶段式升温方式。优化冻干工艺的冻干周期大幅缩短,所得冻干制剂贮存稳定性良好。     

阿霉素脂质体的冻干工艺考察

目的:对阿霉素脂质体冻干工艺进行考察。方法:以外观、再分散性、粒径为指标,对冻干工艺中参数进行优化。结果:优化的冻干工艺为-75℃预冻24h,升温至-25℃保持16h,再升温至20℃持续8h,选择10%海藻糖作为阿霉素脂质体的冻干保护剂。结论:通过优化冻干工艺参数,所得的阿霉素脂质体冻干制品外观平整、质地疏松,加水后能迅速分散均匀。     

冻干过程中常见异常现象的处理

介绍冷冻真空干燥过程中的各种异常现象，分析其原因，提出预防措施，以帮助有关制药单位改善制品的外观，降低冻干制品的废品率和生产成本。     

无盐硫酸氨基葡萄糖冻干工艺优化

目的:为冻干无盐硫酸氨基葡萄糖寻找最佳工艺条件。方法:采用单因素平行试验法探索硫酸氨基葡萄糖冻干工艺优化条件,计算机编制升温程序、升温曲线,调整冻结时间、控制升华速率。优化的试验条件是冷凝器表面温度-45℃、预冷开始工作温度-31℃、液料厚度3cm、预冻时间6h;真空冷冻干燥的搁板表面温度10℃、最终板温35℃;升华界面温度与冷凝器冰表面保持温差7～12℃。结论:①冻结速率对冻结时间、升华时间及能源消耗的影响非常显著,冻结采用缓慢冻结法有利于升华的进行,冻干效果较好;②物料厚度对升华干燥时间影响显著。     

纯化人白细胞α-干扰素热稳定性试验的研究

为探讨纯化人白细胞α-干扰素的热稳定性,确定其有效期,我们用细胞病变抑制法(微量法)。将三批冻干制品分别放置在37℃、22℃、4℃、-20℃定期测定其效价,计算效价损失率结果表明,在不同温度,相同时间内,随放置温度升高,效价损失率增加。在同一温度,不同时间内,随放置时间的增长,效价损失率增加。在较低温度4℃及低温-20℃冻干制品效价的损失率增加缓慢,效价相对稳定。     

脂质体冻结的临界参数

本文研究含亲水性药物脂质体,在冻干/再水化过程中改善质量的途径。冻干包括三个步骤,先是产品冻结,以后开始升华(初级干燥),最后次级干燥。为了在再水化后保持最高的药物量,在三个步骤中均需要最佳的操作参数。作者以亲水而不与双分子层相互作用的羧基荧光素(CF)为模型药物,以氢化大豆磷脂和双十六烷基磷酸酯等材料,用经典薄膜法制备负电荷多层脂质体。并研究冻结速度、温度、时间(样品保持于选定温度的时间)以及不同类型和浓度对冻结保护剂的影响。     

冻干过程中喷瓶现象浅析

喷瓶是影响冻干制剂外观质量的重要因素。本文就造成喷瓶的原因进行了分析,消除药液中的气泡,适宜的预冻温度(约低于共晶点15℃)及控制好冻干速率可有效地消除喷瓶现象。     

注射用还原型谷胱甘肽冻干工艺的优化

为提高注射用还原型谷胱甘肽的稳定性及缩短冻干周期,以冻千时间、产品外观、水分等关键质量属性为指标,对溶剂和干燥温度进行了单因素优化.最终确定采用1％叔丁醇的水溶液作溶剂溶解谷胱甘肽和碳酸氢钠,预冻时隔板温度为-50℃,维持3h至药物全部冻实,再控制一次干燥温度为-36℃,真空度为20 Pa,维持21 h待溶剂完全升华,...     

The improved dissolution and prevention of ampoule breakage attained by the introduction of pretreatment into the production process of the lyophilized formulation of recombinant human Interleukin-11 (rhIL-11)

Lyophilized protein formulations sometimes pose problems such as the formation of a cloudy solution upon reconstitution. Ampoule or vial breakage can also occur during the production processes of lyophilized pharmaceutical products. Various efforts have been made to overcome those difficult problems. In this study, we introduce a particular temperature program into the production process of a recombinant human Interleukin-11 (rhIL-11) lyophilized formulation containing sodium phosphates (Na2HPO4/NaH2PO4, pH 7.0) and glycine in an attempt to improve its dissolution properties and to prevent ampoule breakage from occurring. The formulation was pretreated by nucleating ice and maintaining the solution overnight at a temperature of -6 degrees C. The solution was then completely frozen at a lower temperature. This pretreatment proved successful in not only producing a lyophilized cake which readily disintegrated and dissolved in the reconstitution media, but also prevented ampoule breakage from occurring during the production processes. In contrast, a lyophilized cake produced without the pretreatment created a cloudy solution particularly when reconstituted using water for injection contaminated with aluminum (Al3+), although the solution became transparent within 20-30 min. The pretreatment induced the crystallization of sodium dibasic phosphate (Na2HPO4) in the freeze-concentrate whereas direct freezing without the pretreatment did not crystallize the salt. Thermal analyses (DSC and TMA) showed that amorphous sodium dibasic phosphate in the freeze-concentrate became crystallized upon heating, accompanied by an increase in volume, which probably caused the ampoule breakage that occurred without the pretreatment. Although power X-ray diffraction (PXRD) experiments suggested that, with or without the pretreatment, glycine assumed the beta-form and sodium phosphate stayed amorphous in the final products, an electrostatic interaction between dibasic phosphate anions and rhIL-11, a highly cationic protein, would only exist in the lyophilized cake produced without the pretreatment. This interaction is highly likely because aluminum facilitates the formation of a cloudy solution upon reconstitution possibly by using the divalent anions which effectively reduce electrostatic repulsions between aluminum and the protein to form an aggregate structure that is not readily soluble. The pretreatment would circumvent the interaction by crystallizing the sodium salt before freezing creating a relatively soluble lyophilized cake that is much less sensitive to aluminum.     

Effects of benzyl alcohol on aggregation of recombinant human interleukin-1-receptor antagonist in reconstituted lyophilized formulations

A major limitation in the successful development of multidose protein formulations is protein aggregation induced by antimicrobial preservatives such as benzyl alcohol, which are included to maintain product sterility. Studies were conducted to evaluate the strategy of developing lyophilized formulations of a therapeutic protein, recombinant human interlukin-1 receptor antagonist (rhIL-1ra), to be reconstituted with a bacteriostatic amount (0.9% w/v) of benzyl alcohol in water. The strategy was based on the following hypotheses. The first was that benzyl alcohol would foster aggregation during reconstitution of the lyophilized sample. The second hypothesis was that the extent of benzyl alcohol-induced protein aggregation would correlate directly with the degree of structural perturbation of rhIL-1ra in the dried solid after lyophilization. Differential structural retention of rhIL-1ra in the dried solid was obtained by using a combination of formulation variables important for lyophilization and included: protein concentration, type of stabilizer, and presence or absence of NaCl. Infrared spectroscopic analysis of the lyophilized samples indicated that high initial solution protein concentration and the stabilizer sucrose minimized structural perturbation of rhIL-1ra during lyophilization. In contrast, NaCl was destabilizing. Reconstitution of the dried solid with 0.9% (w/v) benzyl alcohol caused a greater degree of protein aggregation than reconstitution with water, confirming our first hypothesis. In support of our second hypothesis, the extent of aggregation induced by benzyl alcohol during reconstitution was strongly modulated by the degree of retention of native rhIL-1ra secondary structure during lyophilization. During storage of the reconstituted lyophilized samples at room temperature, benzyl alcohol did not accelerate aggregation of rhIL-1ra. This study demonstrated that for development a multidose lyophilized protein formulation involving reconstitution with a solution of benzyl alcohol, protein structural perturbations during freeze-drying should be minimized with a stabilizing excipient and appropriate choice of protein concentration and tonicity modifier. Furthermore, postreconstitution storage at reduced temperature (e.g., room temperature or 4 degrees C) could minimize the risk of preservative-induced protein aggregation.     

Optimizing storage stability of lyophilized recombinant human interleukin-11 with disaccharide/hydroxyethyl starch mixtures

Optimal storage stability of a protein in a dry formulation depends on the storage temperature relative to the glass transition temperature (T(g)) of the dried formulation and the structure of the dried protein. We tested the hypothesis that optimizing both protein structure and T(g)--by freeze-drying recombinant human interleukin-11 (rhIL-11) with mixtures of disaccharides and hydroxyethyl starch (HES)--would result in increased storage stability compared with the protein lyophilized with either disaccharide or hydroxyethyl starch alone. The secondary structure of the protein in the dried solid was analyzed immediately after lyophilization and after storage at elevated temperatures by infrared spectroscopy. After rehydration, aggregation was monitored by size exclusion chromatography. Oxidation levels and cleavage products were quantified by reversed-phase chromatography. For the formulation with HES alone, which has a relatively high T(g), storage stability of rhIL-11 was poor, because HES failed to inhibit lyophilization-induced unfolding. The sugar formulations inhibited unfolding, and had intermediate T(g) values and storage stabilities. Addition of hydroxyethyl starch to sucrose or trehalose increased T(g) without affecting the capacity of the sugar to inhibit protein unfolding during lyophilization. Optimal storage stability of lyophilized rhIL-11 was achieved by using a mixture of disaccharide and polymeric carbohydrates.     

Development of a lyophilization formulation that preserves the biological activity of the platelet-inducing cytokine interleukin-11 at low concentrations

Recombinant human interleukin-11 (rhIL-11) is a licensed biological therapeutic product in at least one country and is used to combat thrombocytopenia during chemotherapeutic regimens, as well as undergoing clinical trials for a range of other disorders. Following attempts to lyophilize IL-11 at low concentrations, it was clear that a significant loss of recoverable biological activity occurred. Investigation of a variety of factors, including the type of container in which the rhIL-11 was lyophilized, revealed that surface adsorption to glass was a major factor resulting in loss of activity of rhIL-11 in solution (> 40% reduction after 3 h at room temperature), in addition to losses of activity post-lyophilization. To overcome this problem, different formulations containing combinations of human serum albumin (HSA), trehalose and Tween-20 have been investigated. Two formulations were successful in entirely preserving the biological activity of rhIL-11 through lyophilization and subsequent reconstitution (potency estimates of formulated relative to original material being > or =0.97). Accelerated degradation studies, performed at intervals over a six-month period, demonstrated the stability of freeze-dried rhIL-11 using these formulations (predicted annual reduction in potency after storage at -20 degrees C < or =1.4%). In conclusion, we have developed a working combination of excipients (0.5% HSA, 0.1% trehalose and 0.02% Tween-20 in potassium phosphate buffer (pH 7.4)) to formulate a stable rhIL-11 freeze-dried product in glass containers, with no loss in potency. These findings should facilitate development of low dose rhIL-11 products and be an indicator of caution to those using this and other material with similar physical properties, without taking appropriate precautions to avoid losses through adsorption.     

温度对采用HPLC法精制白细胞介素-2的影响

目的分析在不同柱温状态下HPLC法对制备rhIL-2收率的影响。方法将rhIL-2复性液以50ml.min-1的速度进样,以不同的乙腈浓度进行洗脱,以280nm紫外分光光度法检测分析,并分析在不同的柱温状态下HPLC法对制备rhIL-2的收率影响。结果与结论采用HPLC法制备重组人白细胞介素-2时,有效的控制温度有利于rhIL-2收率,当柱温为24℃—28℃时蛋白收率较高,纯度符合要求。     

Rapid Depressurization Based Controlled Ice Nucleation in Pharmaceutical Freeze-drying: The Roles of the Ballast Gas and the Vial

Controlled ice nucleation offers several key benefits to the pharmaceutical lyophilization process, including reducing lyophilization cycle time, control of ice crystal morphology, and increased consistency of lyophilized product quality attributes. The rapid depressurization based controlled ice nucleation technique is one of the several demonstrated controlled ice nucleation technologies and relies on the rapid discharge of an inert pressurized gas to induce ice nucleation. In this work, a series of custom wireless gas pressure and temperature sensors were developed and applied to this process to better understand the mechanism of controlled ice nucleation by depressurization. The devices capture highly transient conditions both in the chamber near the vial and within the vial headspace throughout the entire process. The effects of ballast gas composition, initial charge pressure, and vial size on gas pressure and headspace/chamber temperature are explored individually. We model the depressurization as an isentropic process, allowing the influence of these parameters to be evaluated quantitatively. It is demonstrated that monatomic gases (e.g. argon) with low thermal conductivity produce the most favorable conditions for ice nucleation at the end of depressurization, based on temperature drop in the vial headspace. Experimental data also reveal a correlation between initial charge pressure and vial size with the temperature drop within the vial headspace, during depressurization. These findings ultimately provide deeper insight into the rapid depressurization based controlled ice nucleation process and help lay the foundation for a more robust process development and control.     

Improved Folding Yields of a Model Protein Using Protein Disulfide Isomerase

Purpose. To study the effects of recombinant human protein disulfide isomerase (rhPDI) concentration, reduced glutathione:oxidized glutathione ratio (GSH:GSSG) and temperature on the efficiency of oxidative folding of a model protein, recombinant human interleukin 2 (C125A mutation) (C125A rhIL-2). Methods. C125A rhIL-2 inclusion bodies were reduced and denatured by guanidium hydrochloride (Gdm.Cl) and 100 mM GSH. The solution was diluted 10 times into folding buffer, allowing C125A rhIL-2 to fold either in the absence or presence of rhPDI. The renatured and unfolded C125A rhIL-2 species were quantitated by reversed phase-HPLC. Results. The initial folding rate of C125A rhIL-2 linearly increased with rhPDI:C125A rhIL-2 molar ratio in the first 2.5 minutes, and reached the highest rate when the rhPDI:C125A rhIL-2 ratio was 1:1. The oxidative folding of C125A rhIL-2 linearly increased as the GSH:GSSG molar ratio decreased from 10:0 to 10:3. The folding of C125 A rhIL-2 was also dependent on temperature, and optimum folding was realized at 23°C. Conclusions. These results demonstrate that under optimal redox potential and temperature, rhPDI enhances the oxidative folding of C125A rhIL-2. In the oxidative folding of C125A rhIL-2, rhPDI exerts its effect on folding by the acceleration of thiol/disulfide interchange.     

盐酸普鲁卡因注射液稳定性考察

目的考察温度和湿度对普鲁卡因注射液的稳定性影响。方法取3批供试品,在温度(40±2)℃、相对湿度(75±5)%的条件下,分别取放置0、2、4、6个月的样品。用高效液相色谱法测定盐酸普鲁卡因注射液中盐酸普鲁卡因和其降解产物对氨基苯甲酸的含量。结果普鲁卡因注射液中盐酸普鲁卡因的含量随时间延长逐渐下降,6个月下降了10.42%,降解产物对氨基苯甲酸随时间延长逐渐增加,6个月增加了412.89%。结论普鲁卡因注射液在温度(40±2)℃和相对湿度(75±5)%条件下不稳定。