Lyophilization provides long-term stability for a lipid nanoparticle-formulated,nucleoside-modified mRNA vaccine

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叶酸受体靶向β-榄香烯固体脂质纳米粒的制备及其大鼠体内分布

 目的研究叶酸复合物修饰β-榄香烯固体脂质纳米粒(SLN)制剂在大鼠体内的药动学与组织分布情况。方法脂质中加入聚乙二醇单甲醚胆固醇琥珀酸酯(CHS-PEG)和N-硬脂酰基-N′-蝶酰谷氨酰基-聚乙二醇二胺(FA-PEG-S),采用超声-挤压过滤法制备叶酸受体靶向β-榄香烯SLN制剂(FA-PEG-SLN)并加以表征。大鼠尾静脉给药,HPLC测定该制剂的药动学特征及组织分布情况,与普通β-榄香烯SLN(SLN-1)及榄香烯注射乳剂的相比较。结果静脉给药后,FA-PEG-SLN在血浆中的消除半衰期为44.0min,显著长于SLN-1和乳剂的15.6和15.4min。与SLN-1相比,FA-PEG-SLN5min时在肝肾中的药物浓度较高,在脾中稍低。30和60min时,FA-PEG-SLN组在除肺外的各组织中的β-榄香烯浓度皆显著高于SLN-1组和乳剂组。结论FA-PEG-SLN能够在血浆中较长时间循环,在主要器官中的浓度较高且消除速度较慢,有进一步研究的价值。     

Multivariate optimization of mebeverine analysis using molecularly imprinted polymer electrochemical sensor based on silver nanoparticles

Thin film of a moleculary imprinted polymer (MIP) based on electropolymerization method with sensitive and selective binding sites for mebeverine (MEB) was developed. This film was cast on pencil graphite electrode (PGE) by electrochemical polymerization in solution of pyrrole (PY) and template MEB via cyclic voltammetry scans and further electrodeposition of silver nanoparticles (AgNPs). Several parameters controlling the performance of the silver nano particles MIP pencil graphite electrode (AgNPs-MIP-PGE) including concentration of PY(mM) concentration of mebeverine (mM), number of cycles in electropolymerization, scan rate of CV process (mV. s⁻¹), deposition time of AgNPs on to the MIP surface (s), stirring rate of loading solution (rpm), electrode loading time (min), pH of Britton–Robinson Buffer (BRB) solution were examined and optimized using multivariate optimization methods such as Plackett–Burman design (PBD) and central composite design (CCD). Two dynamic linear ranges of concentration for the MIP sensor were obtained as. 1 × 10⁻⁸ to 1 × 10⁻⁶ and 1 × 10⁻⁵ to 1 × 10⁻³ M with the limit of detection (LOD) of 8.6 × 10⁻⁹ M (S/N = 3). The proposed method was successfully intended for the determination of MEB in real samples (serum, capsule). The sensor was showed highly reproducible response (RSD 1.1%) to MEB concentration.     

Pulmonary delivery of siRNA against acute lung injury/acute respiratory distress syndrome

The use of small interfering RNAs (siRNAs) has been under investigation for the treatment of several unmet medical needs, including acute lung injury/acute respiratory distress syndrome (ALI/ARDS) wherein siRNA may be implemented to modify the expression of pro-inflammatory cytokines and chemokines at the mRNA level. The properties such as clear anatomy, accessibility, and relatively low enzyme activity make the lung a good target for local siRNA therapy. However, the translation of siRNA is restricted by the inefficient delivery of siRNA therapeutics to the target cells due to the properties of naked siRNA. Thus, this review will focus on the various delivery systems that can be used and the different barriers that need to be surmounted for the development of stable inhalable siRNA formulations for human use before siRNA therapeutics for ALI/ARDS become available in the clinic.     

星点设计-效应面法优化淫羊藿苷固体脂质纳米粒制备工艺

目的采用星点设计-效应面法优化淫羊藿苷固体脂质纳米粒制备工艺。方法采用高压乳匀法制备淫羊藿苷固体脂质纳米粒,考察卵磷脂,F68,投药量对包封率,载药量以及药物利用率的影响,应用星点设计-效应面法优化处方工艺。结果采用二项式方程拟合实验结果,相关性较好(r>0.9),采用优化后处方条件对预测值进行验证,包封率为(93.09±0.13)%,载药量为(6.34±0.18)%,药物利用率为(84.53±2.45)%,与预测值偏差较小。结论采用星点设计-效应面法优化淫羊藿苷固体脂质纳米粒处方工艺,快速简单,预测准确度高,是较为理想的处方设计及优化方法。     

氟尿嘧啶炭纳米粒新型制剂的淋巴靶向性研究

目的对氟尿嘧啶(5-FU)炭纳米粒新型制剂在大鼠体内的淋巴靶向性进行研究。方法采用反相高效液相色谱法测定大鼠经腹腔注射5-FU炭纳米粒新剂型与5-FU普通剂型(20mg/kg体重)后大鼠体内淋巴组织的药物浓度。结果腹腔注射5-FU炭纳米粒新剂型组中淋巴组织中药物浓度高于5-FU普通剂型组,炭纳米粒可以吸附5-FU并被淋巴组织吞噬,体现出一定的淋巴靶向性,5-FU炭纳米粒新剂型组中的5-FU在淋巴组织中持续时间较长。结论 5-FU炭纳米粒注射液能显著改善淋巴靶器官局部药物浓度,具有较好的淋巴靶向性。     

纳米磁小体氟尿嘧啶微球靶向治疗肝癌的实验研究

目的评价纳米磁性氟尿嘧啶微球治疗肝癌的靶向效应。方法肝癌裸鼠模型32只，随机分成4组：实验组，采用自制的0．03T强度的磁性支架丝，在肿瘤内部建立磁场，尾静脉注射氟尿嘧啶纳米磁小体；生理盐水对照组，无磁场和药物应用；单纯内磁场组，建立0．03T肿瘤局部内磁场，无药物治疗；单纯氟尿嘧啶治疗组，尾静脉注射氟尿嘧啶注射液，无磁场应用。各组于治疗前及连续5d治疗完成后第1，4，7，10，13天各用游标卡尺测量肿瘤大小。电镜观察肿瘤组织病理变化。结果实验组肿瘤抑制率64．0％，与其他三组的肿瘤体积有显著性差异（P〈0．05），该组肿瘤组织镜下显示大量细胞凋亡。结论纳米磁性氟尿嘧啶微球在内磁场的作用下有明显的靶向治疗效应。     

星点设计-效应面法优化pH值依赖型岩黄连碱口服结肠靶向纳米粒

目的 采用星点设计-效应面法（central composite design-response surface methodology,CCD-RSM）优化pH值依赖型岩黄连碱口服结肠靶向纳米粒（dehydrocavidine-chitosan/pectin-nanoparticles,DC-CS/PT-NPs）制备工艺,并对其进行质量表征及体外释放行为评价。方法 采用离子凝胶法制备DC-CS/PT-NPs,以粒径、PDI、ζ电位、包封率、载药量作为评价指标,采用单因素考察和CCD-RSM优化DC-CS/PT-NPs制备工艺。通过透射电子显微镜（transmission electron microscope,TEM）、扫描电子显微镜（scanning electron microscope,SEM）、傅里叶红外光谱（Fourier transform infrared spectroscopy,FT-IR）、差示扫描量热法（differential scanning calorimetry,DSC）和X射线衍射法（X-ray diffraction,XRD）对DC-CS/PT-NPs进行表征,并进行体外释放性能评价。结果 最佳处方为壳聚糖质量浓度为1.5 mg/mL,果胶质量浓度为1.5 mg/mL,TPP质量浓度为2.0 mg/mL,壳聚糖pH值为5.0。DC-CS/PT-NPs包封率为（61.64±1.77）%,载药量为（8.05±0.18）%,粒径为（418.65±4.92）nm,ζ电位为（−14.14±0.22）mV。DC-CS/PT-NPs呈均匀的球形或类球形;制备成纳米粒后,药物的晶型发生了改变;体外释药结果表明,DC-CS/PT-NPs在人工胃液中2 h仅释放24.35%,在人工小肠液中4 h累积释放率＜40%,在人工结肠液中10 h累积释放率＞85%。结论 CCD-RSM所建立的模型可用于DC-CS/PT-NPs处方优化,DC-CS/PT- NPs具有良好的体外结肠释药特征。     

Plasmon Tuning of Liquid Gallium Nanoparticles through Surface Anodization

In this work, tunable plasmonic liquid gallium nanoparticles (Ga NPs) were prepared through surface anodizing of the particles. Shape deformation of the Ga NPs accompanied with dimpled surface topographies could be induced during electrochemical anodization, and the formation of the anodic oxide shell helps maintain the resulting change in the particle shape. The nanoscale dimple-like textures led to changes in the localized surface plasmon resonance (LSPR) wavelength. A maximal LSPR red-shift of ~77 nm was preliminarily achieved using an anodization voltage of 0.7 V. The experimental results showed that an increase in the oxide shell thickness yielded a negligible difference in the observed LSPR, and finite-difference time-domain (FDTD) simulations also suggested that the LSPR tunability was primarily determined by the shape of the deformed particles. The extent of particle deformation could be adjusted in a very short period of anodization time (~7 s), which offers an efficient way to tune the LSPR response of Ga NPs.     

Antibodies to Crucial Epitopes on HSV-2 Glycoprotein D as a Guide to Dosing an mRNA Genital Herpes Vaccine

The toxicity of mRNA-lipid nanoparticle (LNP) vaccines depends on the total mRNA-LNP dose. We established that the maximum tolerated dose of our trivalent mRNA-LNP genital herpes vaccine was 10 μg/immunization in mice. We then evaluated one of the mRNAs, gD2 mRNA-LNP, to determine how much of the 10 μg total dose to assign to this immunogen. We immunized mice with 0.3, 1.0, 3.0, or 10 μg of gD2 mRNA-LNP and measured serum IgG ELISA, neutralizing antibodies, and antibodies to six crucial gD2 epitopes involved in virus entry and spread. Antibodies to crucial gD2 epitopes peaked at 1 μg, while ELISA and neutralizing titers continued to increase at higher doses. The epitope results suggested no immunologic benefit above 1 μg of gD2 mRNA-LNP, while ELISA and neutralizing titers indicated higher doses may be useful. We challenged the gD2 mRNA-immunized mice intravaginally with HSV-2. The 1-μg dose provided total protection, confirming the epitope studies, and supported assigning less than one-third of the trivalent vaccine maximum dose of 10 μg to gD2 mRNA-LNP. Epitope mapping as performed in mice can also be accomplished in phase 1 human trials to help select the optimum dose of each immunogen in a multivalent vaccine.