Method and apparatus for improving in vitro measurement of...

Chemistry: analytical and immunological testing – Optical result – With fluorescence or luminescence

Reexamination Certificate

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C436S171000, C436S050000

Reexamination Certificate

active

07022528

ABSTRACT:
This invention improves the PAMPA (parallel artificial membrane permeability assay) method used in pharmaceutical, biotechnological, and agrochemical R&D. This new high-throughput method and apparatus for measurement of permeability and membrane retention of compounds overcomes shortcomings of prior art and includes sensitivity and speed enhancing reagents. The phospholipid membranes used here consist of 10–74% wt/vol soybean lecithin extract dissolved in dodecane. Concentrations are measured by direct UV spectroscopy. To reduce membrane retention, surfactants, cyclodextrins, or water-soluble lipophilic polymers with low UV absorption are used in the acceptor comparment of the permeation cells and create an artificial sink state. A pH gradient established between donor and acceptor solutions creates a secondary sink state. This “double-sink” makes successful modeling of passive-diffusion transport of molecules possible. By accelerating transport of certain molecules, it shortens measurement time and increases assay throughput. A new permeability equation accounts for the “double-sink” condition as well as membrane retention.

REFERENCES:
patent: 4812407 (1989-03-01), Buchmann et al.
patent: 2002/0004244 (2002-01-01), Avdeef et al.
patent: 101 18 725 (2002-10-01), None
patent: WO 03/003007 (2003-01-01), None
Drug absorption in vitro model: filter-immobilized artificial membranes 2. Studies of the permeability properties of lactones in Piper methysticum Forst; Avdeef et al., European Journal of Pharmaceutical Sciences, www.elsevier.nl/locate/ejps, pp. 271-280.
Physicochemical Profiling(Solubility, Permeability and Charge State); Alex Avdeef, Current Topics in Medicinal Chemistry 2001, Bentham Science Publishers Ltd., pp. 277-351.
Physicochemical High Throughput Screening: Parallel Artificial Membrane Permeation Assay in the Description of Passive Absorption Processes; Manfred Kansy, et al, Journal of Medicinal Chemistry, vol. 41, No. 7, Mar. 26, 1998, pp. 1007-1010.
High-Throughput Permeability pH Profile and High-Throughput Alkane/Water log P with Artificial Membranes; Frank Wohnsland, et al., Novartis Pharma AG, WKL-122.P.33, CH-4002 Basel, Switzerland, Journal of Medicinal Chemistry, 2001, vol. 44, pp. 923-930.
Optimized conditions of bio-mimetic artificial membrane permeation assay; Kiyohiko Sugano, et al.; International Journal of Pharmaceutics 228 (2001), pp. 181-188.
A comparative study of artificial membrane permeability assay for high throughput profiling of drug absorption potential; Chengyue Zhu, et al., European Journal of Medicinal Chemistry 37 (2002), pp. 399-407.
*Solid-Supported Lipid Membranes as a Tool for Determination of Membrane Affinity: High-Throughput Screening Of a Physicochemical Parameter; A. Loidl-Stahlhofen et al.; Journal of Pharmaceutical Sciences, vol. 90, No. 5, May 2001.
*Physiocochemical Profiling in Drug Research: A Brief Survey of the State-of-the-Art of Experimental Techniques; A. Avdeef et al.; CMLS Cellular and Molecular Life Sciences59 (2002) pp. 1681-1689.

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