Immobilized artificial membranes IAM

A very promising method, immobilized artificial membrane (IAM) chromatography, was developed by Pidgeon and co-workers [299-304,307], where silica resin was modified by covalent attachment of phospholipid-like groups to the surface. The retention parameters mimic the partitioning of drugs into phospholipid bilayers. The topic has been widely reviewed [47,298,307,309-311]. 

Ottiger, C. Wunderli-Allenspach, H., Immobilized artificial membrane (IAM)-HPLC for partition studies of neutral and ionized acids and bases in comparison with the liposomal partition system, Pharm. Res. 16, 643-650 (1999). 

Taillardat-Bertschinger, A. Martinet, C. A. M. Carrupt, P.-A. Reist, M. Caron, G. Fmttero, R. Testa, B., Molecular factors influencing retention on immobilized artificial membranes (IAM) compared to partitioning in liposomes and n-octanol, Pharm. Res. 19, 129-Til (2002). 

Immobilized artificial membranes (IAM) are another means of measuring lipophilic characteristics of drug candidates and other chemicals [90-94], IAM columns may better mimic membrane interactions than the isotropic octanol/water or other solvent/solvent partitioning system. These chromatographic indices appear to be a significant predictor of passive absorption through the rat intestine [95]. 

Such new RP-HPLC stationary-phase materials have been available for some years (Regis Chemical Company, Morton Grove, IL, USA). These so-called immobilized artificial membrane (IAM) columns consist of lipid molecules covalently bound to propylamine-silica. The unreacted propylamine moieties are end-capped with methylglycolate. The membrane lipid, phosphatidylcholine, possesses polar head groups and two non-polar hydrocarbon chains (C18). One of the alkyl chains is linked to the propylamine-silica surface. 

Correlation to HAS and IAM column retentivity VD is a function of albumin binding and partitioning into phospholipid membranes, and these can be measured using affinity HPLC columns Retentivity on human albumin and immobilized artificial membrane HPLC columns [33]... 

Due to their better biomimetic properties, phospholipids have been proposed as an alternative to 1-octanol for lipophiiicity studies. The use of immobilized artificial membranes (lAM) in lipophiiicity determination was recently reviewed and we thus only briefly summarize the main conclusions [108]. lAM phases are silica-based columns with phospholipids bounded covalently. lAM are based on phosphatidylcholine (PC) linked to a silica propylamine surface. Most lipophiiicity studies with lAM were carried out using an aqueous mobile phase with pH values from 7.0 to 7.4 (log D measurements). Therefore, tested compounds were neutral, totally or partially ionized in these conditions. It was shown that the lipophiiicity parameters obtained on I AM stationary phases and the partition coefficients in 1-octanol/water system were governed by different balance of intermolecular interactions [109]. Therefore the relationships between log kiAM and log Poet varied with the class of compounds studied [110]. However, it was shown that, for neutral compounds with log Poet > 1, a correspondence existed between the two parameters when double-chain lAM phases (i.e., lAM.PC.MG and IAM.PC.DD2) were used [111]. In contrast, in the case of ionized compounds, retention on lAM columns and partitioning in 1 -octanol / water system were significantly different due to ionic interactions expressed in lAM retention but not in 1-octanol/water system and due to acidic and basic compounds behaving differently in these two systems. 

Support for these results comes from a paper in which the partitioning of 17 dmgs into adipose tissue was examined [84]. For these drugs, log Poct, log Doct atpH 74, and the retention time on an immobilized artificial membrane, log k IAM, were determined. The adipose storage index (ASI) was defined as ... 

The use of partition coefficients between water and lipophilic media is of wide use in pharmaceutical research. As discussed in the last chapters, different lipophilicity scales are used to describe the lipophilicity of a compound and relate it to its absorption behaviour in vivo. Differences between the logPow and partitioning between phospholipids and water (mainly determined using liposomes) for diverse compounds have been described leading to the development of the immobilized artificial membrane chromatography system. However, also the predictivity of the IAM system is limited and only a small number of membrane systems are available. 

Immobilized Artificial Membrane (LAM) packings for HPLC provide a different environment from that of the hydrocarbon-based ODS columns [17,30—32]. In this model, IAMs are purified phospholipids that are covalently bonded to the silicon support. At this time, only IAM columns prepared from phosphatidylcholine are commercially available (Regis Technologies, Inc., Morton Grove, IL). The hypothesis is that the bonded phospholipid layer is akin to the cellular monolayer that represents a barrier to transport. Interaction of the solute with the phospholipid results in a capacity factor ( J that is proportional to the membrane partition coefficient [PCm in Eq. (1)]. In this sense, the IAM approach does not attempt to correlate with literature values of oil water partition coefficients, but seeks to establish a unique membrane interaction parameter. 

Figure 9.4 Immobilized artificial membrane lAM) chromatography. Binding of solute to fluid membranes in (A) can be modeled by solute binding to an immobilized membrane as in (B) through measurement of iam as shown in (C). (Reprinted with permission from Yang, C.Y., el al. Immobilized artificial membranes—Screens for drug membrane interactions. Adv. Drug Deliv. Rev. 1996, 23 229-256, copyright 1997, Elsevier).

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