Development of topical nanoemulsion product containing Benjakul remedy extract for anti-inflammation

Benjakul is a Thai traditional remedy and used to be adaptogen. It iscomposed with five plants such as Piper chaba Hunt., Piper sarmentosum Roxb.,Piper interruptum Opiz., Plumbago indica Linn., and Zingiber officinale Roscoe. It hasever been reported to treat Osteoartitis knee patients. It showed good efficacyequally NSAID drug (Diclofenac). However it has side effect such as stomach pain.Thus, the aim of this research was to develop to be a topical nanoemulsion productcontaining BKE for anti-inflammation, to reduce side-effect and to increase theefficient delivery of the drug.The ethanolic extract of Benjakul before adding to the NE-base showedanti-inflammatory activity with IC50 value of 19.72 ± 0.72 µg/ml. and it had piperinecontent of 88.89 mg/g. Stress testing found that the extract was treated underthermolysis, hydrolysis, acid hydrolysis, base hydrolysis, and oxidation condition hadanti-inflammatory activity with IC50 values of 21.17±2.09, 15.97±1.4, 19.47±3.2,20.87±3.2, 15.35±1.8 µg/ml respectively and piperine content in Benjakul extract ofevery conditions were not decreased significantly. In the development ofnanoemulsion the solubility of piperine in BKE was studied by dissolving in oleic acid,Ref. code: 25615811031524RMU(2)myritol, isopropyl myristate, oil mix and tween80. The results showed that piperinecontent in every solvent were 31.00, 34.16, 41.41, 44.86, 51.30 mg/g, respectively.The development of a topical nanoemulsion product containing BKEinvolved the study of the various formulations, i.e. oleic acid, myritol, isopropylmyristate, tween80, and other compositions with properties in the acceptable rangewere selected for the study. BKE3-NE-T8-x0.2 contains BKE and oil mixed (oleic acid:myritol: isopropyl myristate), and aqueous phase (propylene glycol, paraben cocn.)contains the surfactant (tween80), water and xanthan gum. Properties of the bestformula of Benjakul nanoemulsion (BKE3-NE-T8-x0.2) was tested and showedparticle size, PDI, zeta potential, and viscosity as 587.6 ± 25.0 nm, 0.6 ± 0.09 and -5.7± 0.2 mV respectively. The viscosity had a flow curve exhibiting non-Newtonian flowbehavior (pseudoplastic) and the release of BKE from nanoemulsion was 0.0855µg/min.BKE3-NE-T8-x0.2 showed anti-inflammatory activity by inhibitory effecton nitric oxide induced by LPS in RAW264.7 cells, with IC50 of 8.36±1.0 µg/ml. andcytotoxicity activity by using MTT assay in HaCaT cells showed that the percentage ofcell viability was more than 70% at every concentration which indicated that theywere not toxic to human skin. The stability test of BKE3-NE-T8-x0.2 indicated it wasunstable but still anti-inflammatory and piperine content was not reduced,surprisingly BKE3-NE-T8-x0.2 showed increasing anti-inflammatory activity indicatingthat it can be stored more than two years.In conclusion, BKE3-NE-T8-x0.2 is an effective anti-inflammatory productand it is also a non-toxic on keratinocytec cell line. Although, the product wasunstable but it still showed higher anti-inflammatory activity when it was keep longtime. Therefore, these results can conclude that a topical nanoemulsion productcontaining BKE has potential as an anti-inflammatory drug. The further researchBKE3-NE-T8-x0.2 should be studied on anti-inflammatory activities and, irritation testin the animal model and normal volunteers for safety.