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Development of novel cationic microemulsion as parenteral adjuvant for influenza vaccine

Asian Journal of Pharmaceutical Sciences 2020年第5期15卷 591-604页

作者： Sakalanunt Lamaisakul Angkana Tantituvanont vimolmas lipipun Garnpimol Ritthidej Department of Pharmaceutics and Industrial Pharmacy Faculty of Pharmaceutical SciencesChulalongkorn UniversityBangkok 10330Thailand Department of Biochemistry and Microbiology Faculty of Pharmaceutical SciencesChulalongkorn UniversityBangkok 10330Thailand

Squalene-based oil-in-water(O/W)emulsions have been used as effective and safe adjuvants in approved influenza vaccines.However,there are concerns regarding the safety and side effects of increasing risk of narcolepsy.In present study,novel O/W microemulsions(MEs)containing wheat germ oil,D-alpha tocopheryl polyethylene glycol 1000 succinate(TPGS)and Cremophor EL(CreEL)or Solutol HS15 were formulated with/without a cationic surfactant,cetyltrimethylammonium bromide(CTAB)and then sterilized by autoclaving.Their physical properties and biological efficacies were evaluated.The results demonstrated that autoclaving reduced the droplet size to^20 nm with narrow size distributions resulting in monodisperse systems with good stability up to 3 years.Hemolytic activity,viscosity,pH,and osmolality were appropriate for parenteral use.Bovine serum albumin(BSA),a model antigen,after mixing with MEs retained the protein integrity,assessed by SDS-PAGE and CD spectroscopy.Greater percentages of 28 SC cell viability were observed from CreEL-based MEs.Uptake of FITC-BSA-MEs increased with the increasing concentration of CTAB confirmed by CLSM images.Furthermore,cationic CreEL-based MEs could induce Th1 cytokine synthesis with an increase in TNF-α and IL-12 levels and a decrease in IL-10 level.In vivo immunization study in mice of adjuvants admixed with influenza virus solution revealed that nonionic and selected cationic CreEL-MEs enhanced immune responses as measured by influenza-specific serum antibody titers and hemagglutination inhibition titers.Particularly,cationic CreEL-based ME showed better humoral and cellular immunity with higher IgG2 a titer than nonionic CreEL-based ME and antigen alone.No differences in immune responses were observed between mice immunized with selected cationic CreEL-based ME and marketed adjuvant.In addition,the selected ME induced antigen-sparing while retained immune stimulating effects compared to antigen alone.No inflammatory change in muscle fiber structure was observed.Accordingly,the develop

关键词： Cationic microemulsion Adjuvant Influenza virus Vaccine

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Development of polyurethane foam dressing containing silver and asiaticoside for healing of dermal wound

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Asian Journal of Pharmaceutical Sciences 2019年第1期14卷 40-51页

作者： Nantaporn Namviriyachote vimolmas lipipun Yada Akkhawattanangkul Phingphol Charoonrut Garnpimol C.Ritthidej Faculty of Pharmaceutical Sciences Chulalongkorn UniversityBangkok 10330Thailand Faculty of Veterinary Science Mahidol UniversityNakhon Pathom 73170Thailand

Polyurethane foam dressings for dermal wounds were formulated with natural polyols in order to improve the foam characteristics and the release of 2 active agents,silver and asiaticoside(AS)as an antimicrobial agent and an herbal wound healing agent,respectively.The foam was instantly formed by interaction of polyols and diisocyanate.Hydroxypropyl methylcellulose,chitosan and sodium alginate were individually mixed with themain polyols,polypropylene glycol,in the formulation while the active componentswere impregnated into the obtained foam dressing sheets.Although the type and amount of the natural polyols slightly affected the pore size,water sorption-desorption profile and compression strength of the obtained foam sheets,a prominent effect was found in the release of both active components.Among natural polyols formulations,foam sheets with alginate showed the highest silver and AS release.Non-cytotoxicity of these foam sheets to human fibroblast cells was confirmed.Antimicrobial testing on four bacteria strains showed that 1mg/cm^2 silver in formulations with 6%of natural polyols and without natural polyols had sufficient content of the silver release with comparable inhibition zone and significantly larger zone than other formulations.In pig study,the foam dressing with 6%alginate,1mg/cm^2 silver and 5%AS could improve wound healing in both the percentage of the wound closure and histological parameters of the dermal wound without any dermatologic reactions.In conclusion,this innovative foam dressing had potential to be a good candidate for wound treatment.

关键词： Polyurethane foam dressing Natural polyols Silver nanoparticles Asiaticoside Wound healing

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The identities and anti-herpes simplex virus activity of Clinacanthus nutans and Clinacanthus siamensis

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Asian Pacific Journal of Tropical Biomedicine 2013年第4期3卷 284-290页

作者： Paween Kunsorn Nijsiri Ruangrungsi vimolmas lipipun Ariya Khanboon Kanchana Rungsihirunrat College of Public Health Sciences Chulalongkorn UniversityBangkok 10330Thailand Faculty of Pharmaceutical Sciences Chulalongkorn UniversityBangkok 10330Thailand

Objective:To distinguish the difference among the Clinacanthus nutans(Burm.f.)Lindau(C.nutans)and Clinacanthus siamensis Bremek(C.siamensis)by assessing pharmacognosy characteristics,molecular aspect and also to evaluate their anti-herpes simplex virus(HSV)type 1 and type 2 activities.Methods:Macroscopic and microscopic evaluation were performed according to WHO Geneva guideline.Stomatal number,stomatal index and palisade ratio of leaves were evaluated.Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PGR and then sequenced.Dry leaves were subsequently extracted with n-hexane,dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay.Results:Cross section of midrib and stem showed similar major components.Leaf measurement index of stomatal number,stomatal index and palisade ratio of C.nutans were 168.32±29.49,13.83±0.86 and 6.84±0.66,respectively,while C.siamensis were 161.60±18.04,11.93±0.81and 3.37±0.31,respectively.The PCR amplification of ITS region generated the PGR product approximately 700 bp in size.There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions.The IC_(50)values of C.nutans extracted with n-hexane,dichloromethane and methanol against HSV-1 were(32.05±3.63)μg/mL,(44.50±2.66)μg/mL,(64.93±7.00)μg/mL,respectively where as those of C.siamensis were(60.00±11.61)μg/mL,(55.69+4.41)μg/mL,(37.39±5.85)μg/mL,respectively.Anti HSV-2 activity of n-hexane,dichloromethane and methanol C.nutans leaves extracts were(72.62±12.60)μg/mL,(65.19±21.45)μg/mL,(65.13±2.22)μg/mL,respectively where as those of C.siamensis were(46.52±4.08)μg/mL,(49.63±2.59)μg/mL,(72.64±6.52)μg/mL,respectively.Conclusions:The combination of macroscopic,microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.

关键词： Clinacanthus nutans Clinacanthus siamensis Microscopic analysis Biomolecular analysis Internal transcribed spacer Herpes simplex virus Plaque reduction assay

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Effect of cationic microemulsion on hemolysis and immune cell uptake

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Asian Journal of Pharmaceutical Sciences 2016年第1期11卷 164-165页

作者： Sakalanunt Lamaisakul Angkana Tantituvanont vimolmas lipipun Garnpimol C.Ritthidej Department of Pharmaceutics and Industrial Pharmacy Chulalongkorn UniversityBangkok 10330Thailand Department of Biochemistry and Microbiology Faculty of Pharmaceutical SciencesChulalongkorn UniversityBangkok 10330Thailand

Squalene-based oil-in-water nanoemulsions, constituents in influenza vaccines, have been approved in Europe as human licensed adjuvants (1)With similar constituents but easier manufacturing procedure and greater physical stability [2],microemulsion was of interest. To improve cellular uptake,positive surface charge of nanosystems has been reported(3)In the present study, cationic microemulsions were prepared, characterized and evaluated in immune cell.

关键词： Cationic microemulsion Hemolysis Adjuvant Immune cell uptake

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