Transcellular Route

There are two routes potentially involved in drug absorption across the nasal epithelial barrier the transcellular and paracellular routes [20], Several experimental evidences dealing with the mechanism of transnasal permeation support the existence of both lipoidal pathyway (i.e., transcellular route) and an aqueous pore pathway (i.e., paracellular route). 

This refers to the transport across the epithelial cells, which can occur by passive diffusion, carrier-mediated transport, and/or endocytic processes (e.g., transcytosis). Traditionally, the transcellular route of nasal mucosa has been simply viewed as primarily crossing the lipoidal barrier, in which the absorption of a drug is determined by the magnitude of its partition coefficient and molecular size. However, several investigators have reported the lack of linear correlation between penetrant lipophilicity and permeability [9], which implies that cell membranes of nasal epithelium cannot be regarded as a simple lipoidal barrier. Recently, compounds whose transport could not be fully explained by passive simple diffusion have been investigated to test if they could be utilized as specific substrates for various transporters which have been identified in the 

The existence of a carrier-mediated transport in nasal mucosa was first suggested by Kimura et al. [34], P-glycoprotein, organic cation transporter, dopamine transporter, and amino acid transporters have all been identified in the nasal mucosa, especially in the olfactory mucosa [31, 32, 35, 36], These transporters determine the polarized absorption and excretion of their substrates including amino acids, amines, and cations. 

Since the uptake of particles in nasal epithelial tissue is known to be mostly mediated by M cells, nasal administration has been investigated as a noninva-sive delivery of vaccines [37], However, since the uptake of naked DNA by endocytocis is limited, use of either nanoparticles as mucosal delivery systems [37] or hypotonic shock [38] is reported for the efficient transfection of gene and vaccine into the nasal epithelium. It was also reported that polypeptides and polypeptide-coated nanospheres (diameter about 500 nm) are transported through endocytic process in rat M cells [39], 

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Kelder et al. [19] have shown that PSA can be used to model oral absorption and brain penetration of drugs that are transported by the transcellular route. A good correlation was found between brain penetration and PSA (n=45, r=0.917). From analyzing a set of 2366 central nervous system (CNS) and non-CNS oral drugs that have reached at least phase 11 clinical trials it was concluded that orally active drugs that are transported passively by the transcellular route should have PSA< 120 Al In addition, different PSA distributions were found for CNS and non-CNS drugs. 

Most drugs appear to be absorbed in humans by passive diffusion (linear or first-order kinetics). The predominant pathway taken by most drugs is through the epithelial cell, the transcellular route. It is this route that requires the compound to have a reasonable K0/w... 

The perturbation of monolayers with agents (e.g., disodium ethylenediamine tetraacetate, Ca+2-free medium, sodium citrate, cytochalasin D) to open tight junctions and the effect on the transmonolayer flux of permeants are addressed in this section. It has been observed that permeants taking predominantly the trans-cellular route are not affected by perturbants of the paracellular route, compared to extracellular or relatively hydrophilic permeants (Artursson and Magnusson, 1990). Let us put these general observations into a quantitative intepretation in the light of the transmonolayer kinetic studies of steroids in this section and of paracellular permeants in Section III. There are three cases to consider (1) ABL-controlled permeants, (2) monolayer-controlled permeants transported principally by the transcellular route, and (3) monolayer-controlled permeants for which the paracellular route dominates. 

If the paracellular route is perturbed to the extent that the pore radius R is changed from 12 A to 500 A, then F(5/500) 1.0 and Pparacell = 6.1 x 10 6 cm/sec. Although this represents a 12-fold gain from 5 X 10 7 cm/sec, the transcellular route taken by testosterone is yet the dominant pathway of the monolayer. In making the final calculations, we can conclude that the change in the observed / , will be imperceptible. 

Case 2 Monolayer-Controlled Permeants by the Transcellular Route... 

There are two principal routes of drug transport across any epithelium transcellu-lar and paracellular (Fig. 6). In the transcellular route, drugs are transported... 

Figure 6 Epithelial penetration routes for topically applied drugs. The transcellular route (1) is preferred by lipophilic drugs, while the paracellular route (2) is preferred by hydrophilic drugs.

For the evaluation of a possible relationship between the molecular structure of a potential candidate and its transport abilities to cross the epithelial membrane of the gut, the mechanism or route of transport must be known [1,4]. This is due to the structural requirements for the transcellular route being different from the paracellular route. During the lead optimization phase - when many mechanistically based studies are performed - the cell culture-based models can also be used with great confidence. 

Molecules with a large molecular weight or size are confined to the transcellular route and its requirements related to the hydrophobicity of the molecule. The transcellular pathway has been evaluated for many years and is thought to be the main route of absorption of many drugs, both with respect to carrier-mediated transport and passive diffusion. The most well-known requirement for the passive part of this route is hydrophobicity, and a relationship between permeability coefficients across cell monolayers such as the Caco-2 versus log P and log D 7.4 or 6.5 have been established [102, 117]. However, this relationship appears to be nonlinear and reaches a plateau at around log P of 2, while higher lipophilicities result in reduced permeability [102, 117, 118]. Because of this, much more attention has recently been paid towards molecular descriptors other than lipophilicity [86, 119-125] (see section 5.5.6.). The relative contribution between the para-cellular and transcellular components has also been evaluated using Caco-2 cells, and for a variety of compounds with different charges [110, 112] and sizes [112] (see Section 5.4.5). 

Originating from the structure of the stratum corneum, two permeation pathways are possible (a) the intercellular route and (b) the transcellular route. 

Under normal conditions, the transcellular route is not considered as the preferred way of dermal invasion, the reason being the very low permeability through the corneocytes and the obligation to partition several times from the more hydrophilic corneocytes into the lipid intercellular layers in the stratum corneum and vice versa. The transcellular pathway can gain in importance when a penetration enhancer is used, for example, urea, which increases the permeability of the corneocytes by altering the keratin structure. 

Secretory epithelia control transport of water and solutes from the subluminal compartment (blood) into the lumen or body exterior. At present, there is no single unifying model for transepithelial fluid or water transport. In some epithelia, transcellular routes of fluid transport via water channels may predominate [88a], However, in other types of epithelia, such as the cervical-vaginal epithelia, transport of fluids usually occurs via the paracellular route [1, 14], In the latter, movement of fluid can be driven by three main mechanisms (Figure 15.1C) ... 

Orally active drugs that are transported passively by the transcellular route can be tailored to brain permeation by decreasing the PSA to less than 60-70 A2. 

Recent years have witnessed an explosive growth in the imderstanding of the mechanisms associated with the absorption of drugs, especially therapeutic peptides and proteins. Scientists from a variety of disciplines continue to elucidate the variables associated with the optimal formulation and delivery of drugs via the oral mucosa. A greater rmderstanding of the para- and transcellular route of drug absorption, pro-... 

Compounds can cross biological membranes by two passive processes, transcellu-lar and paracellular mechanisms. For transcellular diffusion two potential mechanisms exist. The compound can distribute into the lipid core of the membrane and diffuse within the membrane to the basolateral side. Alternatively, the solute may diffuse across the apical cell membrane and enter the cytoplasm before exiting across the basolateral membrane. Because both processes involve diffusion through the lipid core of the membrane the physicochemistry of the compound is important. Paracellular absorption involves the passage of the compound through the aqueous-filled pores. Clearly in principle many compounds can be absorbed by this route but the process is invariably slower than the transcellular route (surface area of pores versus surface area of the membrane) and is very dependent on molecular size due to the finite dimensions of the aqueous pores. 

For simple molecules, like p-adrenoceptor antagonists octanol/water log D74 values are remarkably predictive of absorption potential. Compounds with log Dy 4 values below 0 are absorbed predominantly by the paracellular route and compounds with log Dy 4 values above 0 are absorbed by the transcellular route. 

The use of in vitro models for prediction of compounds that are predominantly absorbed passively by the transcellular route is generally good with these models. Predicting compounds which are absorbed paracellularly or via active uptake or efflux mechanisms is more difficult. There is a lack of understanding of expression levels of transporters in the gut, which makes in vivo predictions difficult. 

Beside membrane transporters such as PepTl and PepT2, which act as absorptive systems, there are transporters like P-gp and the MRP 15, which transport certain drugs actively back into the intestinal lumen. These efflux pumps are located in several tissues including liver, kidney, brain, and intestine [90,91]. In the intestine, efflux systems are predominantly located at the apical side of the epithelial cells. Lipophilic drugs are usually absorbed by the transcellular route so that they are mostly affected by these systems. Interestingly, the intracellular occurring CYP3A metabolizes compounds to substrates that are eliminated by P-gp [92],... 

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