Research On The Anti-migraine Mechanism Of Zanthoxylum Schinifolium Volatile Oil And The Preparation Of Patch

Zanthoxylum schinifolium（ZS）is the dry mature pericarp of Zanthoxylum schinifolium Sieb.et Zucc.which belongs to Rutaceae,together with Red pepper,which is collectively referred to as Zanthoxylum bungeanum（ZB）.It is a characteristic spice and traditional Chinese medicine resource in southwest China,which has the effect of dispelling cold and relieving arthralgia,killing insects and relieving itching."Theory of Medicinal Properties"records:ZB cures“head wind”and tears.The"head wind"of traditional Chinese medicine includes migraine in modern medicine,and there is also a folk use of prickly ash to treat migraine.In the herbal literature,ZB is mainly used to relieve pain,and modern pharmacological studies have also confirmed its analgesic effect,and the analgesic effect and duration of ZS is higher than that of ZB,and its toxicity is lower than that of ZB,but there are few studies on the anti-migraine effect of ZS.Based on this,our research group studied the efficacy and mechanism of Zanthoxylum schinifolium Essential oil（ZSEO）in the treatment of migraine,and planned to develop it into a hot melt pressure sensitive adhesive transdermal patch for migraine relief,making it a transdermal drug delivery product that is easy to give,easy to expose,quick to take effect,safe and non-toxic,and to promote the development of traditional Chinese medicine resources with southwest characteristics and the innovation of migraine prevention and treatment products.Chemical composition research:Five kinds of ZSEO,were prepared by CO₂supercritical fluid extraction（SFE-CO₂）,steam distillation（SD）,Soxhlet extraction（SE）,ultrasonic extraction（UE）and simultaneous distillation extraction（SDE）.The composition was analyzed by gas chromatography-mass spectrometry（GC-MS）and the relative abundance of each compound was calculated.The extraction rates of different extraction methods and the characteristics of different volatile oils were compared,and the best extraction method was selected.The results showed that the yield of ZSEO extracted by SD and SFE-CO₂was 7.6%and 12.3%,respectively.There were 23 components in the volatile oil obtained by SD,16 species of SE,13species of UE and 13 species of SDE.The main components of ZSEO were linalool,limonene and sabinene.Considering the extraction rate,fluidity and the demand of large-scale industrial production,the volatile oil was prepared by SFE-CO2 method and the follow-up study on efficacy and preparation was carried out.Study on the anti-migraine activity of ZSEO:The classic nitroglycerin（NTG）migraine rat model and reserpine migraine mouse model were selected to evaluate the anti-migraine effect of ZSEO and explore its mechanism.The results show that ZSEO can alleviate the migraine behavior of animals induced by nitroglycerin and reserpine,reduce the up-regulation of NO,ET-1 and CGRP in animals,increase the expression of 5-HT,and reduce NF-κBp65,IκBαand IL-1βin TCC,suggesting that ZSEO may reduce neurogenic inflammation by affecting vasomotor and inhibiting NF-κB pathway,and play an anti-migraine effect.Study on the anti-migraine activity of linalool:Linalool is the main component of ZSEO,and its relative abundance is more than 70%.The NTG migraine rat model was used to explore whether linalool is the main active component of ZSEO against migraine.It was found that 185 mg kg^-1linalool could reduce the increase of head scratching times in migraine rats induced by NTG,and increase the expression of NO,ET-1 and CGRP in serum,which was the main active component of ZSEO against migraine.In addition,transdermal linalool had the best effect,followed by olfactory administration,and oral administration had the worst effect.Preparation of ZSEO Hot Melt pressure sensitive adhesive:According to the sensory evaluation of homogeneity,spreadability,permeation degree,skin following,exfoliation,repeated exfoliation and residual,as well as initial adhesion,adhesion holding force and peeling strength as indexes,the ratio of hot melt pressure sensitive adhesive matrix was screened by single factor design and Box-Behnken design-response surface method,the in vivo animal evaluation method was used to screen the number of flexure heads of NTG migraine rats as index to select the dosage of the paste.The results show that the optimum process of patch is SIS elastomer 8.0g,hydrogenated petroleum resin 4.0 g,liquid paraffin 10.0 g,and the optimum drug loading per patch is 70 mg.Safety investigation of ZSEO hot melt pressure sensitive adhesive:The safety of ZSEO hot melt pressure sensitive adhesive was evaluated by acute toxicity test in mice,skin irritation test in rabbits and skin hypersensitivity test in guinea pigs.The results showed that ZSEO hot melt pressure sensitive adhesive had no acute toxicity to mice,no allergy to guinea pig skin and mild irritation to rabbit skin,but it was eliminated within 6 hours,which proved that ZSEO hot melt pressure sensitive adhesive could be used safely.