Development of a dog microdialysis model for determining synovial fluid pharmacokinetics of anti-arthritis compounds exemplified by methotrexate.
OBJECTIVE
The purpose of this study was to develop and validate an animal model of drug disposition in synovial fluid (SF) by comparing microdialysis with arthrocentesis using the anti-arthritic drug methotrexate (MTX).


METHODS
Microdialysis probes were calibrated in vitro with the no net flux method using dog synovial fluid. The probes were implanted surgically into the stifle joint space of four dogs and were dialyzed overnight using a portable microinfusion pump. The membrane integrity of the probes was monitored by retrodialysis using an internal standard. After an intravenous bolus of 2.5 mg/kg of MTX, unbound concentrations in synovial fluid, as well as total plasma concentrations, were measured by liquid chromatography tandam mass spectrometer (LC/MS/MS) in samples collected from 0 to 48 h postdose.


RESULTS
The probe membrane remained intact at least 48 h after implantation. The mean probe recovery and unbound fraction of MTX in SF were 46.8% and 44.8%, respectively. The unbound fraction of MTX was 44% in synovial fluid. MTX penetrated into the joint space rapidly, with maximal concentrations of 6.6 microM reached at approximately 1 h postdose. The unbound MTX area under the curve in SF was approximately 40% of the total area under the curve in plasma. These data agree well with the previous data obtained for MTX using arthrocentesis.


CONCLUSIONS
In contrast with arthrocentesis, microdialysis enables the collection of multiple serial SF samples from individual animals with minimal trauma and potential blood contamination. This animal model should prove valuable for studying the disposition of new antiarthritis compounds or biomarkers in SF.
Qian
Mingxin
M
Department of Drug Metabolism and Pharmacokinetics, Bristol Myers Squibb Company, Wilmington, Delaware, USA. mingxin.qian@bms.com
West
Wanda
W
Wu
Jing-Tao
JT
Lu
Bing
B
Christ
David D
DD
D000818:Animals; D018501:Antirheumatic Agents / Q000493:pharmacokinetics*; D004285:Dogs; D005240:Feasibility Studies; D008401:Gas Chromatography-Mass Spectrometry; D008297:Male; D008727:Methotrexate / Q000737:chemistry / Q000493:pharmacokinetics*; D017551:Microdialysis; D023421:Models, Animal; D019152:Paracentesis; D013582:Synovial Fluid / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism*
D018501:Antirheumatic Agents; D008727:Methotrexate