Permeation-enhancing effects of chitosan formulations on recombinant hirudin-2 by nasal delivery in vitro and in vivo.
OBJECTIVE
To investigate the enhancing effects of chitosan with or without enhancers on nasal recombinant hirudin-2 (rHV2) delivery in vitro and in vivo, and to evaluate the ciliotoxicity of these formulations.


METHODS
The permeation-enhancing effect of various chitosan formulations was estimated by using the permeation coefficient of fluorescein isothiocyanate recombinant hirudin-2 (FITC-rHV2) across the excited rabbit nasal epithelium in vitro. The effect was further evaluated by measuring the blood concentration level after nasal absorption of FITC-rHV2 in rats. The mucosal ciliotoxicity of different formulations was evaluated with an in situ toad palate model.


RESULTS
Chitosan at a concentration of 0.5% with or without various enhancers significantly increased the permeability coefficient (P) and relative bioavailability (Fr) of FITC-rHV2 compared with the blank control. The addition of 1% sodium dodecylsulfate, 5% Brij35, 5% Tween 80, 1.5% menthol, 1% glycyrrhizic acid monoammonium salt (GAM) or 4% Azone into the 0.5% chitosan solution resulted in a further increase in absorption (P<0.05) compared with 0.5% chitosan alone. But co-administration of chitosan with 5% hydroxyl-propyl-beta-cyclodextrin (HP-beta-CD), 5% lecithin or 0.1% ethylenediamine tetraacetic acid (EDTA) was not more effective than using the 0.5% chitosan solution alone. Chitosan alone and with 5% HP-beta-CD, 0.1% EDTA, 1% GAM or 5% Tween 80 was relatively less ciliotoxic.


CONCLUSIONS
Chitosan with or without some enhancers was able to effectively promote the nasal absorption of recombinant hirudin, while not resulting in severe mucosal ciliotoxicity. A chitosan formulation system would be a useful approach for the nasal delivery of recombinant hirudin.
Zhang
Yu-jie
YJ
School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100083, China.
Ma
Chang-hua
CH
Lu
Wan-liang
WL
Zhang
Xuan
X
Wang
Xiao-liang
XL
Sun
Jian-ning
JN
Zhang
Qiang
Q
D000042:Absorption / Q000187:drug effects; D000281:Administration, Intranasal; D000818:Animals; D019540:Area Under Curve; D001682:Biological Availability; D002023:Bufo bufo; D048271:Chitosan / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D016650:Fluorescein-5-isothiocyanate / Q000493:pharmacokinetics*; D006629:Hirudins / Q000493:pharmacokinetics*; D008297:Male; D009297:Nasal Mucosa / Q000187:drug effects*; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000493:pharmacokinetics
D006629:Hirudins; D011994:Recombinant Proteins; C072299:hirudin HV2; D048271:Chitosan; D016650:Fluorescein-5-isothiocyanate