[Consideration about chemistry, manufacture and control (CMC) key problems in simplified registration of classical traditional Chinese medicine excellent prescriptions].

As an outstanding representative of traditional Chinese medicine(TCM) prescriptions accumulated from famous TCM doctors' clinical experiences in past dynasties, classical TCM excellent prescriptions (cTCMeP) are the most valuable part of TCM system. To support the research and development of cTCMeP, a series of regulations and measures were issued to encourage its simplified registration. There is still a long-way to go because many key problems and puzzles about technology, registration and administration in cTCMeP R&D process are not resolved. Based on the analysis of registration and management regulations of botanical drug products in FDA of USA and Japan, and EMA of Europe, the possible key problems and countermeasures in chemistry, manufacture and control (CMC) of simplified registration of cTCMeP were analyzed on the consideration of its actual situation. The method of "reference decoction extract by traditional prescription" (RDETP) was firstly proposed as standard to evaluate the quality and preparation uniformity between the new developing product under simplified registration and traditional original usages of cTCMeP, instead of Standard Decoction method in Japan. "Totality of the evidence" approach, mass balance and bioassay/biological assay of cTCMeP were emphatically suggested to introduce to the quality uniformity evaluation system in the raw drug material, drug substance and final product between the modern product and traditional decoction.

CuI/1,10-phen/PEG promoted decarboxylation of 2,3-diarylacrylic acids: synthesis of stilbenes under neutral and microwave conditions with an in situ generated recyclable catalyst.

A series of trans- or cis-stilbenes have been synthesized in good to excellent yields via a functional group-dependent decarboxylation process from the corresponding 2,3-diaryl acrylic acids in a neutral CuI/1,10-phen/PEG-400 system under microwave conditions. The in situ generation of the recyclable catalytic complex, the use of environmentally benign solvent PEG-400, the operational simplicity, the short reaction times, as well as the functional group-dependent chemo- and stereo-selectivity have made the decarboxylation process a highly efficient and applicable protocol.

Binaprofen induces zebrafish liver injury via the mitochondrial pathway.

Binaprofen (C18H23NO5) is a drug not commercially available that causes liver injury; however, the underlying mechanism is unknown. The aim of the present study was to determine the mechanism underlying binaprofen­induced liver injury at the genetic level. Zebrafish were treated with binaprofen. Serum biomarkers [alanine transaminase (ALT), aspartate transaminase (AST) and lactate dehydrogenase (LDH)], malondialdehyde (MDA) and glutathione (GSH) content analysis, liver cell morphology examination, DAPI staining, electron microscopy, microarray analysis and reverse transcription­quantitative (RT­q)PCR were performed 12, 24 and 48 h post­treatment to analyze the mechanism underlying binaprofen­induced liver injury. Following exposure to binaprofen, zebrafish serum levels of ALT, AST and LDH increased; MDA content of liver tissue increased and GSH content decreased. Liver cells exhibited mild to moderate vacuolization and mitochondria exhibited vacuolization and disrupted cristae. Liver cell apoptosis rate increased. There were 190 common differentially expressed genes at 12, 24 and 48 h. Gene Ontology analysis showed that the function of downregulated genes was primarily associated with 'DNA replication', 'DNA metabolic process', 'cell cycle', 'cell redox homeostasis', 'mitochondrion' and 'lipid transport'. The function of upregulated genes was primarily associated with 'peroxisome proliferator', 'oxidation activity', 'peroxisome' and 'apoptosis'. Pathway analysis showed that downregulated genes were those pertaining to 'cell cycle', 'DNA replication', 'ribosome', 'spliceosome', 'pyrimidine metabolism', 'purine metabolism', upregulated genes were those pertaining to 'PPAR signaling pathway', 'p53 signaling pathway'; RT­qPCR assay supported the microarray results. The mechanism underlying binaprofen­induced liver injury was associated with lipid peroxidation and apoptosis. Binaprofen downregulated genes associated with lipid transport and anti­apoptosis genes, upregulated pro­apoptosis genes and induces liver cell injury via the mitochondrial signaling pathway.

Cross-linked thermosensitive nanohydrogels for ocular drug delivery with a prolonged residence time and enhanced bioavailability.

AIM: A temperature-triggered, cross-linked nano hydrogel formulation (NPs-gel) was prepared to prolong the residence time of dexamethasone (DXM) in the eye and increase its bioavailability. RESEARCH DESIGN AND METHODS: The NPs-gel was prepared by combining a high pressure homogenization method with a cold solution method. Soy lecithin E200, lecithin oil, glycerol, kolliphor P188, kolliphor P407, and polycarbophil were the excipients used for the formation of NPs-gel containing DXM. The nanoparticle size, temperature-sensitive phase transition characteristics, in vitro and in vivo release behavior, corneal permeability, and eye irritation level of the NPs-gel were evaluated. RESULTS: The NPs-gel had slightly larger particle size than the DXM-loaded nanoparticles, yet it retained the properties of nanoparticles such as surface effect and size effect. The phase transition temperature was 33.2 °C, which is within the trigger conditions of intraocular temperature. Under physiological conditions, the adhesion and adhesion work of the NPs-gel were 1.1 and 2.1 times that of an in situ-formed gel, and the gel strength of NPs-gel was 1.8 times that of an in situ-formed gel. These results indicate that NPs-gel has greater adhesion and mechanical strength. The area under the curve of NPs-gel was 3.08 and 1.51 times that of DXM-loaded nanoparticles and in situ-formed gel, showing higher bioavailability. CONCLUSION: The NPs-gel is a suitable formulation to further enhance ocular drug delivery.

Effects of serum and follicular fluid on the in vitro maturation of canine oocytes.

The effects of gonadotropin, serum and follicular fluid on the in vitro maturation of canine oocytes were examined. Additionally, spindle size and spindle migration in MI-stage oocytes derived by in vivo or in vitro maturation were evaluated for the first time. Mature oocytes collected from beagle dog ovaries were divided into two experiments. In experiment I, oocytes were cultured in basic TCM 199 medium supplemented with different levels of P4, E2 and FSH. In experiment II, oocytes in the estrus or anestrus stage were cultured in basic medium supplemented with 30% or 40% canine serum plus 20% or 10% follicular fluid. Our results showed that in experiment I, more oocytes reached MI-MII (18.57%) after supplementation with 1 IU/ml FSH+ 5 IU/ml P4 + 5 IU/ml E2 than after supplementation with other levels of reagents. However, there were no significant differences among the groups (three different concentration groups and a control group) with respect to the proportions of oocytes that resumed meiosis, completed meiosis or degenerated. In experiment II, the number of oocytes from the estrus stage that reached MI-MII in TCM 199 medium supplemented with 40% canine serum and 10% follicular fluid (46.72%) was significantly higher (p < 0.01) than the number of oocytes from the anestrus stage that reached MI-MII in medium supplemented with 30% canine serum and 20% follicular fluid (21.84%). In addition, the degeneration rate was significantly lower (p < 0.05) in the 40% canine serum/10% follicular fluid group from follicular stage than in the other three groups. The average spindle length of the MI-stage oocytes that matured in vivo was significantly (p < 0.01) longer than that of the MI-stage oocytes that matured in vitro (21.75 vs. 14.39 µm). These results suggest that supplementation of the culture medium with 40% estrus serum and 10% follicular fluid had a positive influence on the in vitro maturation of canine oocytes and greatly affected spindle size in MI-stage oocytes.

PLGA Nanoparticle Platform for Trans-Ocular Barrier to Enhance Drug Delivery: A Comparative Study Based on the Application of Oligosaccharides in the Outer Membrane of Carriers.

PURPOSE: The trans-ocular barrier is a key factor limiting the therapeutic efficacy of triamcinolone acetonide. We developed a poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs) surface modified respectively with 2-hydroxypropyl-ß-cyclodextrin (2-HP-ß-CD), chitosan oligosaccharide and trehalose. Determination of the drug/nanoparticles interactions, characterization of the nanoparticles, in vivo ocular compatibility tests, comparisons of their corneal permeability and their pharmacokinetics in aqueous humor were carried out. METHODS: All PLGA NPs were prepared by the single emulsion and evaporation method and the drug-nanoparticle interaction was studied. The physiochemical features and in vitro corneal permeability of NPs were characterized while the aqueous humor pharmacokinetics was performed to evaluate in vivo corneal permeability of NPs. Ocular compatibility of NPs was investigated through Draize and histopathological test. RESULTS: The PLGA NPs with lactide/glycolide ratio of 50:50 and small particle size (molecular weight 10 kDa) achieved optimal drug release and corneal permeability. Surface modification with different oligosaccharides resulted in uniform particle sizes and similar drug-nanoparticle interactions, although 2-HP-ß-CD/PLGA NPs showed the highest entrapment efficiency. In vitro evaluation and aqueous humor pharmacokinetics further revealed that 2-HP-ß-CD/PLGA NPs had greater trans-ocular permeation and retention compared to chitosan oligosaccharide/PLGA and trehalose/PLGA NPs. No ocular irritation in vivo was detected after applying modified/unmodified PLGA NPs to rabbit's eyes. CONCLUSION: 2-HP-ß-CD/PLGA NPs are a promising nanoplatform for localized ocular drug delivery through topical administration.