| Objective: To develop a new route of administration, Amiloride hydrochloride has been investigated by transcranial administration in the rat. To study significant differences between transcranial acupuncture and non-acupuncture point administration of amiloride hydrochloride. Transcranial administration is a new administration route for brain diseases treatment, which will make a profound influence in brain diseases treatment.Methods: A HPLC method has been developed and validated for determination of amiloride hydrochloride in rat brain homogenates. Amiloride hydrochloride has been investigated by transcranial administration in the rat, the data were processed by 3p97.The relationship between the thickness of skull and permeability was studied by the study of transcranial administration of amiloride hydrochloride in rat of different ages.To study significant differences between transcranial acupuncture and non-acupuncture point administration of amiloride hydrochloride through the study of transcranial acupuncture point administration of amiloride hydrochloride. Amiloride hydrochloride gel were prepared for the study of transcranial administration in vitro.Eucalyptus oil is selected as penetrant, 3% Azone as controls, then screening penetrant in the study of amiloride hydrochloride of transcranial administration in vitro.Results: Calibration curves equation of Amiloride hydrochloride is y = 70724 x +1983.7. Lnearity of the methods was confirmed(R2=0.9991),over a range of 0.02~0.32μg/m L. The relative standard deviation(RSD) of intra-day precision were3.33%,1.73% and 7.98% respectively in low(0.08 μg/m L), medium(0.16 μg/m L) and high(0.24 μg/m L) concentrations and RSD of inter-day precision were 0.47%,0.72%, 1.04% respectively.All RSD of intra-day and inter-day precision were smaller than 10%, they are corresponding to quality control samples of the method. RSD of stability at ambient laboratory temperature were 0.61%, 0.06%, 1.59% within 10 hours. All of them were smaller than 10%, they are in line with the methodology requirements.The extraction recoveries were 88.2±2.08%,87.9±1.97% and90.4±2.36% respectively in low, medium and high concentrations. All of extraction recoveries were more than or equal to 70%, they are in line with the methodology requirements.The relative recoveries were 105.9±1.12%, 101.2±1.42% and112.0±2.78% in low, medium and high concentrations. All of relative recoveries were between 85% and 115%, they are in line with the methodology requirements.The pharmacokinetics of Amiloride hydrochloride is in line with one-compartment model with first absorption and lag time in rat’s brain(weight=1). The A of Amiloride hydrochloride was 0.266875±0.0290 μg/m L, Ke was 0.004208±0.0002min-1,Ka was0.092585±0.0313 min-1, Lag time was 50.416229±1.3574 min, T1/2(Kα) was7.486592±1.0691 min, T1/2(Ke) was 164.7375564±11.2555 min,Tpeak was85.39389±2.6294 min, Cmax was 0.219884±0.0201 μg/m L, AUC was60.544632±4.0446(μg/m L)*min, CL/F(s) was 1.982009±0.1319 mg/kg/min/(μg/m L),V/F(c) was 471.0563±40.2545(mg/kg)/(μg/m L). The amount of Amiloride hydrochloride were 0.30275μg/m L, 0.22726 μg/m L, 0.06789 μg/m L in brain of young,middle-aged,aged rats. The quatity of amiloride hydrochloride in rat brain after transcranial acupuncture point administration could not be detected by analytical method established.The formulation of amiloride hydrochloride contain 0.4g HPMC, 1.5 g glycerin,then add water to 10 g, adjust p H to 6.Penetration increased after added penetrant, the penetration effects of 4% and 5%eucalyptus oil were better than others. Er were 1.295,1.264. And the penetration effects of 4% and 5% eucalyptus oil is better than 3% Azone. The Q(transdermal) of aminloride hydrochloride is larger than Q(transcranial), but there was no significant difference(P> 0.05).Conclusion: The parameters of amiloride hydrochloride indicated that Amiloride hydrochloride could penetrate into the brain. The penetration of amiloride hydrochloride is inversely proportional to age, which indicated that the penetration of amiloride hydrochloride has a relationship with thickness of skull. The quatity of amiloride hydrochloride in rat brain after transcranial acupuncture point administration could not be detected by analytical method established, indicated that the quatity of amiloride hydrochloride in rat brain was below the detection limit.The work provides a new administration for the treatment of brain diseases. |
PDF Full Text Request