Membrane carrier-mediated

As described above, because MAO is bound to mitochondrial outer membranes, MAOIs first increase the concentration of monoamines in the neuronal cytosol, followed by a secondary increase in the vesicle-bound transmitter. The enlarged vesicular pool will increase exocytotic release of transmitter, while an increase in cytoplasmic monoamines will both reduce carrier-mediated removal of transmitter from the synapse (because the favourable concentration gradient is reduced) and could even lead to net export of transmitter by the membrane transporter. That MAOIs increase the concentration of extracellular monoamines has been confirmed using intracranial microdialysis (Ferrer and Artigas 1994). 

Fig. 9 Schematic representation depicting the movement of molecules from the absorbing (mucosal or apical) surface of the GIT to the basolateral membrane and from there to blood. (A) transcellular movement through the epithelial cell. (B) Paracellular transport via movement between epithelial cells. (Q Specialized carrier-mediated transport into the epithelial cell. (D) Carrier-mediated efflux transport of drug out of the epithelial cell. (Copyright 2000 Saguaro Technical Press, Inc., used with permission.)...

Carrier-mediated transport is linear with mucosal solute concentration until this concentration exceeds the number of available carriers. At this point the maximal solute flux (7max) is independent of further increases in mucosal solute concentration. In the linear range of solute flux versus mucosal concentration (C), the proportionality constant is the ratio of / to the solute-carrier affinity constant (Km). This description of Michaelis-Menten kinetics is directly analogous to time changes in mass per unit volume (velocity of concentration change) found in enzyme kinetics, while here the appropriate description is the time change in solute mass per unit surface area of membrane supporting the carrier. 

Both secondary active transport and positive cooperativity effects enhance carrier-mediated solute flux, in contrast to negative cooperativity and inhibition phenomena, which depress this flux. Most secondary active transport in intestinal epithelia is driven by transmembrane ion gradients in which an inorganic cation is cotransported with the solute (usually a nutrient or inorganic anion). Carriers which translocate more than one solute species in the same direction across the membrane are referred to as cotransporters. Carriers which translocate different solutes in opposite directions across the membrane are called countertransporters or exchangers (Figs. 10 and 11). 

Figure 12 Schematic of generation of mucosal microclimate pH as a transmucosal proton-gradient driving force for di- and tripeptide carrier-mediated translocation across the mucosal membrane into the enterocyte.

The coupled processes described by Eqs. (8), (14), (17), and (22) can be added in (20) as parallel solute transport pathways across the membrane. The phenomenological coefficients (Ly) describe the membrane permeability by these pathways [potential-dependent, Eq. (8) via membrane lipid partition and diffusion, Eq. (14) carrier-mediated, Eq. (17) and convectively coupled, Eq. (22)]. These pathways define parallel resistances through the intestinal barrier in series with precellular resistances to solute transport. 

Solute uptake can also be evaluated in isolated cell suspensions, cell mono-layers, and enterocyte membrane vesicles. In these preparations, uptake is normalized by enzyme activity and/or protein concentration. While the isolation of cells in suspension preparations is an experimentally easy procedure, disruption of cell monolayers causes dedifferentiation and mucosal-to-serosal polarity is lost. While cell monolayers from culture have become a popular drug absorption screening tool, differences in drug metabolism and carrier-mediated absorption [70], export, and paracellular transport may be cell-type- and condition-depen-dent. 

Figure 1 General pathways through which molecules can actively or passively cross a monolayer of cells. (A) Endocytosis of solutes and fusion of the membrane vesicle with the opposite plasma membrane in an active process called transcytosis. (B) Similar to A, but the solute associates with the membrane via specific (e.g., receptor) or nonspecific (e.g., charge) interactions. (C) Passive diffusion between the cells through the paracellular space. (C, C") Passive diffusion (C ) through the cell membranes and cytoplasm or (C") via partitioning into and lateral diffusion within the cell membrane. (D) Active or carrier-mediated transport of an otherwise poorly membrane permeable solute into and/or out of a cellular barrier.

Thus, the fraction of dose absorbed is exponentially related to the absorption number. Equation (10) shows that the absorption number (and therefore the membrane permeability) is a fundamental parameter while other parameters such as the partition coefficient and pKa are useful guides but not fundamental parameters. For highly soluble drugs with linear absorption kinetics, dose and dissolution have no effect on the fraction of dose absorbed. In the case of drugs that are absorbed by a carrier-mediated process, a mean permeability should be used [30],... 

GL Amidon, PJ Sinko, D Fleisher. Estimating human oral fraction dose absorbed A correlation using rat intestinal membrane permeability for passive and carrier-mediated compounds. Pharm Res 5 651-654, 1988. 

Several attempts have been made to estimate the dose required in humans in relation to a drug s potency, and to put this into the context of solubility and permeability for an optimal oral drug [2, 3]. A relatively simple example of this is where a 1.0 mg kg-1 dose is required in humans, then 52 pg mL"1 solubility is needed if the permeability is intermediate (20-80%) [3]. This solubility corresponds approximately to 100 pM of a compound with a MW of 400 g mol-1. Most screening activities for permeability determinations in, e.g., Caco-2, are made at a concentration of 10 pM or lower due to solubility restrictions. The first implication of this is that the required potency for these compounds needs to correspond to a dose of <0.1 mg kg-1 in humans if the drug should be considered orally active. Another implication would be the influence of carrier-mediated transport (uptake or efflux), which is more evident at low concentrations. This could result in low permeability coefficients for compounds interacting with efflux transporters at the intestinal membrane and which could either be saturated or of no clinical relevance at higher concentrations or doses. 

The water-soluble vitamins generally function as cofactors for metabolism enzymes such as those involved in the production of energy from carbohydrates and fats. Their members consist of vitamin C and vitamin B complex which include thiamine, riboflavin (vitamin B2), nicotinic acid, pyridoxine, pantothenic acid, folic acid, cobalamin (vitamin B12), inositol, and biotin. A number of recent publications have demonstrated that vitamin carriers can transport various types of water-soluble vitamins, but the carrier-mediated systems seem negligible for the membrane transport of fat-soluble vitamins such as vitamin A, D, E, and K. 

SlROTNAK, F. M. AND B. TOLNER. Carrier-mediated membrane transport of folates in mammalian cells. Annu. Rev. Nutr. 1999, 39, 91-122. 

Ishizawa, T., et al. Sodium and pH dependent carrier-mediated transport of antibiotic, fosfomydn, in the rat intestinal brush-border membrane. J. Pharmacobiodyn. 1990, 13, 292—300. 

Lastly, pharmacogenomics could provide new tools for the design of more specific and active CNS pharmaceuticals. The efficacy of a broad spectrum of neuro-pharmaceutical drugs is often complicated by their inability to reach their site of action because of the BBB. One way to overcome this is to use carrier-mediated transport at the luminal and/or abluminal membranes of the endothelial cells of the BBB. This will provide a physiologically based drug delivery strategy for the brain by designing new chemical entities or fused proteins that can cross the BBB via these transporters. 

Although the absence of paracellular transport across the BBB impedes the entry of small hydrophilic compounds into the brain, low-molecular-weight lipophilic substances may pass through the endothelial cell membranes and cytosol by passive diffusion [7]. While this physical barrier cannot protect the brain against chemicals, the metabolic barrier formed by the enzymes from the endothelial cell cytosol may transform these chemicals. Compounds transported through the BBB by carrier-mediated systems may also be metabolized. Thus, l-DOPA is transported through the BBB and then decarboxylated to dopamine by the aromatic amino acid decarboxylase [7]. 

Figure 9.4. Schematic representation of carrier-mediated metal-ion transport through a liquid membrane (A = anion).

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