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Advanced Drug Delivery Reviews, 7 (1991) 339 364

(Q 1991 Elsevier Science Publishers B.V. All rights reserved. / 0169-409X/91/$03.50 ADONIS 0169409X91004023 ADR 00103



(B) Mechanisms of Peptide and Protein Absorption (l) Paracellular intestinal transport: modulation of absorption
Hugh N. Nellans
Abbott Laboratories, Department of Pharmacology, G1 Pharm. Sec., Abbott Park, IL, USA
(Received December 12, 1990) (Accepted May 6, 1991)

Key words: Intestinal absorption; Intestinal secretion; Cellular; Paracellular; Permeability; Partition coefficient; Molecular size; Tight junction; Convective flow; Solvent drag

Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I. Intestinal paracellular pathway: functional properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2. Intestinal paracellular pathway: physiological control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . II.Passive modulation of intestinal paracellular transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1. Descriptive equations for transcellular and paracellular fluxes . . . . . . . . . . . . . . . . . . . . . . . 2. Relative transport contributions of transcellular and paracellular pathways ...... 3. Convective flow and paracellular flux . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4. Net intestinal transport as a difference in unidirectional fluxes . . . . . . . . . . . . . . . . . . . . . . 5. Rectified convective water flow: physiological barrier to absorption ............... |II. Active control of paracellular absorption . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .... 1. Mechanism of change in intestinal permeability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2. Other epithelia and in vitro/in vivo comparisons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3. Permeability changes and in vivo absorption . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Abbreviations: PEG, poly(ethylene glycol); MDCK, Madine Darby canine kidney. Correspondence: H.N. Nellans, Abbott Laboratories, Department of Pharmacology (46R), Pharmaceutical Products Division, G1 Pharm. Sec., Building AP9, One Abbott Park Road, Abbott Park, IL 600643500, USA. Fax: (1) (708) 938-5286. 340 340 341 342 344 344 347 348 348 349 352 352 354 354


H.N. N E L L A N S IV. The p a r a c e l l u l a r p a t h w a y in disease states . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . V. P a r a c e l l u l a r probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357 358 359 359 360

VI. E x p l o r a t o r y studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . VII. C o n c l u s i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Summary The significance of the paracellular pathway in both the absorption and secretion of therapeutic compounds by the intestine has received little attention to date. A growing body of literature firmly documents the importance of this aqueous pathway as a major contributor to the transepithelial flow of water and solutes in a wide spectrum of epithelia, including both the small and large intestine. The relative contributions of the paracellular and cellular pathways to intestinal absorption are discussed with respect to both diffusion and convective flow of water. Emphasis is placed on the interaction of convective flow with net intestinal absorption. The discussion of convective flow is extended to the intriguing observations that the permeability of the paracellular tight junction may be under active regulation by glucose and amino acids through cytoskeletal connections to the perijunctional actomyosin complex. The review cites studies from other epithelia, suggesting the universal character of the importance of the transport between, as well as through, cells and attempts to establish physiological criteria for assessing the contribution of paracellular transport to the intestinal absorption of therapeutic compounds, including peptides and peptidomimetics. 1. Introduction Historically, solute transport across biological 'membranes' has been dominated by the concept of diffusion through the lipid barrier of cell plasma membranes. The classic studies of Collander [1], relating membrane permeability to the partition coefficient of an array of non-electrolytes, have served several generations of students and investigators as the appropriate paradigm for considering most biological transport phenomena. However, distinct deficiencies arise within this framework when considering epithelia; it is the objective of this review to point to alternative pathways for transport, particularly those dominated by high water permeation. The specific target will be the pathway between, rather than through, mucosal epithelial cells of the intestinal tract. Despite the fact that a significant body of data identifies the extracellular or paracellular pathway of the intestine as a dominant route for water and electrolyte transport [2-5], many current models of drug absorption

Powell [6] has provided a comprehensive review of the intestinal and gastric barriers to passive diffusion.~ ec~uivalent pore diameter in rat and rabbit [3] and may be slightly smaller (4-8 A).6]. The limiting dimensions of the paracellular pathway under fasting conditions is of the order of 10-15 . however. This is to say that seven times more ion traffic traverses the spaces and intercellular junctions between intestinal epithelial cells than the route through the cells.8] have disregarded the potential role of this pathway in the intestinal absorption of therapeutic drugs. for some electrolytes. The paracellular pathway provides the route for such large volume transport and manipulations of the driving forces and selectivity of this pathway may provide a feasible mode for enhancing oral delivery of small peptides and peptidomimetics. In fact.10]. Cereijido and coworkers [11-13] have been particularly active in the characterization of MDCK (Madine-Darby canine kidney) cells. such as calcium. and factors such as osmotic and . These pivotal studies have established a strong correlation between the electrical conductance of the monolayer and the integrity of the tight junctions. Intestinal paracellular pathway. Cho and coworkers [14. H. In that review he has characterized the role of the tight junction in distinguishing gastrointestinal tissues which transport large quantities of solutes against small gradients (small intestine) from those transporting smaller quantities against large gradients (stomach and colon). The appreciation of this route for drug delivery across the intestine is only beginning to be explored [9. capable of large mass transfer against relatively small gradients of concentration and pressure. it has been recognized that the small intestine of common laboratory mammals. The intent of this review is to discuss the physiological role of the paracellular pathway in intestinal water homeostasis and to explore the implications of control mechanisms for paracellular permeation which may impact oral drug delivery.15] have recently begun more intensive work on the MDCK monolayer to characterize size and charge selectivity of the paracellular pathway. It is primarily the small intestine. Lord and Di Bona [16] have also demonstrated that the septate junctions of invertebrate epithelia serve essentially identical functions to those of vertebrate tight junctions."functional properties For nearly 20 years. as reported by Fordtran and coworkers [17] in man. Virtually all of these kidney epithelial characteristics are shared by the intestine.1. such as rat and rabbit [3.P A R A C E L L U L A R INTESTINAL TRANSPORT: M O D U L A T I O N OF ABSORPTION 341 [7. Cation selectivity has been established and the requisite role of extracellular calcium for maintaining tight junctional structure has been identified. These dimensions are not fixed. the paracellular pathway may be the primary route of intestinal absorption under conditions of adequate intake [4]. although the small intestinal epithelium has not been satisfactorily maintained as an isolated monolayer. which will be the focus of this review. Several reviews have focused on the role of the epithelial tight junction of intact tissue or of cells maintained in cultured monolayers. moves as much as 85% of the transepithelial electrolyte flow via the aqueous environment of the extracellular or paracellular pathway between epithelial cells.

N E L L A N S hydrostatic pressure differences.30] have observed that rat . the dog jejunum increases the equivalent pore dimensions of the convective (paracellular) pathway for water and solutes by at least threefold. in fact. H. For example. totally abolished net water absorption. Extrapolating such observations to homeostatic control of water and solute transport reaffirms that hypertonicity in the serosal space beneath intestinal epithelial cells is capable of inducing water absorption.2. the role of angiotensin II in increasing proximal tubular water reabsorption is well established [24]. Intestinal paracellular pathway: physiological control These observations suggest that both hydrostatic and osmotic forces are able to alter the dimensions of the intercellular space and perhaps the tight junctional complexes. The similarity between intestine and tubule suggests that common mechanisms may. ferritin and Evans blue to access the lumenal solution. It is striking that similar observations have been made in the small intestine where the renin-angiotensin system has been shown to stimulate water absorption [25. These results are entirely consistent with the physical factors known to control water flow in other epithelial such as the renal proximal tubule [21-23].28] and Munck and Rasmussen [29. From the early work of Lifson and his coworkers [18-20]. Such an effect would lead to water conservation similar to that in the kidney. as well as recently described cellular control mechanisms [5. from 10 to 30 A diameter [18-20].32]. Such increases in equivalent pore size are attributable to expansion of the lateral intercellular space and may include changes in dimension of the perijunctional complexes binding adjacent cells to one another. while a reverse tonicity gradient expands the space and increases water absorption.342 H. Further. whereas serosal interstitial hydrostatic pressure and lumenal (mucosal) hypertonicity cause water secretion into the intestine. applied from the serosal side. an effect of angiotensin II to decrease intestinal interstitial pressure could result in increased water absorption. the effect of hydrostatic pressure also alters dimensions of the paracellular pathway in a rectified fashion. it has become clear that mucosal hypertonicity collapses the lateral space between cells and markedly decreases water absorption. It has been known for some time that under conditions of water absorption.26]. Nellans and Kimberg [27. modulate whole body water homeostasis at the two sights. may modulate the limiting dimensions of the paracellular pathway. Other provocative examples of pressure effects on intestinal absorption have been observed with in vitro preparations of small intestine. while pressures as small as 2-6 cm. Differences in response of the paracellular pathway to changes in osmotic and hydrostatic gradients are quite striking and reveal an intriguing degree of asymmetry. Further.N. Although the mechanism has not been established with certainty. Mucosal pressure as high as 22 cm H20 failed to cause any detectable increase in water absorption. increases in serosal pressure to 10 cm H20 led to frank water secretion with corresponding increases in paracellular equivalent dimensions allowing inulin.

Intestinal hydrostatic pressure beneath the mucosal epithelium may also be modulated by influences of intestinal smooth muscle. To date the role of smooth muscle motility in intestinal absorption has been limited to the consideration of mixing and transit time. such as prostaglandins [41] and the renin-angiotensin system [25. pressure builds in the lateral space between cells. displays a remarkable asymmetry in hydrophilic solute transport under conditions which normally stimulate net water absorption. However. poly(ethylene glycol) (PEG) and thiourea. Such water flow contributes to a significant increase in solvent drag of solutes in the serosal to mucosal direction leading to net mannitol.P A R A C E L L U L A R INTESTINAL TRANSPORT: M O D U L A T I O N OF ABSORPTION 343 small intestine. In the absence of sufficient capillary and/or lymphatic outlets for this water. These studies highlight not only the influence of hydrostatic pressure on net intestinal solute flux. have been shown to display net secretory fluxes in the presence of glucose-stimulated ion and water uptake by the tissue. As water uptake into tissue spaces is stimulated by glucose through active ion absorption. may hamper their own intestinal absorption through such a pharmacological mechanism. acting as agonists for smooth muscle contraction. pressure continues rising to a point at which water flows back to the mucosal compartment. in the absence of intact blood supply. This net secretion occurs in the absence of a concentration-dependent difference in driving force for the solutes. Such a pressure increase. The introduction of an influence of motility on paracellular transport of water and contained solutes opens a new arena for interpretation of atypical pharmacokinetics of orally administered compounds. Solutes. It is highly probable that tissue hydrostatic pressure in the intact animal plays a significant role in water and water-driven solute absorption (or secretion) through the paraceilular pathway. may be capable of either inhibiting absorption through the paracellular pathway and/or causing secretion of absorbed solutes residing in the subepithelial space. if of sufficient magnitude. but also raise the cautionary note that in vitro transport studies must be continually compared to in vivo counterparts to ascertain the relevance of observations from isolated tissues to those in the intact animal. remains to be delineated and will be discussed in greater detail in subsequent sections. Either or both effects would reduce the net intestinal absorption of lumenal solutes as well as the systemic bioavailability. PEG and thiourea fluxes (see Fig. Limited observations from this laboratory suggest that erythromycin derivatives. such pharmacological interactions must be considered when evaluating anomalies in intestinal absorption for compounds which may influence intestinal tissue water flow through effects on intestinal blood pressure. These passive pressure effects. together with the active control of . 2).26]. It is suggested that increased intestinal motility may contribute to either transient or sustained intestinal tissue pressure. The influence of physiological factors controlling such tissue pressure. such as mannitol. presumably through the junctional complexes of the paracellular pathway. The explanation for this anomaly lies in the generation of hydrostatic pressure within the subepithelial space of these isolated intestinal preparations.

N E L L A N S paracellular pathway permeability. As an initial attempt to consider the relative contributions of both pathways to absorption and secretion.1.. The cellular and paracellular pathways for solute and water flux (J) across the intestinal epithelium are depicted using two adjacent cells with the tight junctional complex illustrated by broken lines. II. 1. provide the focus for the following discussions..N. In this model. The fundamental assumption for developing a formalism for intestinal transport is that both absorption (mucosal to serosal flux) and secretion (serosal to mucosal flux) may occur through the parallel pathways of the columnar epithelial cells and the junctions and associated lateral spaces between the cells. Passive modulation of intestinal paraeellular transport H.344 H. Descriptive equations for transcellular and paracellularfluxes A comprehensive view of intestinal transport in the current era demands inclusion of both cellular and extracellular (paracellular) routes of translocation. Parallel pathways for intestinal absorption and secretion.. . as depicted in Fig. Paracellular Pathway Fig. 1. the following set of descriptive flux equations is advanced to account for mass transfer without consideration of the small effects of epithelial potential on fluxes of solutes with low charge to mass ratios. as well as the aqueous/membranous route through the cellular cytosol. the resistance to transcellular flux is considered to include both the apical and basolateral plasma membranes. Intestinal Epithelial Cell Lumen __~ Jcell ~Spacer_ Subepithelial JparaceII .

For initial approximation. respectively. multiplied by the diffusional driving force (concentration in compartment of origin. Specifically. LIS) may also contribute impedance to flow. the transport path lengths for the two pathways are considered equivalent. If the driving forces for solute flux across the intestinal epithelium are initially limited to diffusion and convection. In the case of the transcellular flux. C . both the transcellular and paracellular pathways are described with unidirectional diffusional flux equations. This solvent drag phenomenon is completely ignored in transport treatments which do not . At this point. cm 2) of the relevant pathway. For all uncharged compounds and most charged compounds with relatively low charge to mass ratios. Jv represents the volume or convective water flux (#l. the above equations would be sufficient to describe mass transfer or flux (tool/h) in the unidirectional dimension. Although some bulk water transport (convection) may occur through the cells.l) and will be considered INTESTINAL TRANSPORT: MODULATION OF ABSORPTION 345 The primary resistance to paracellular flux is considered to reside in the tight junctional complexes [3. the partition coefficient is considered to be unity. however.6.32]. AceH ~cell (1) Jparacell = ( Dparacell+ DparacellJvl " C DH20 (2) (2)The diffusion coefficients (D. it is considered insignificant relative to the mass transfer between the cells. For the paracellular pathway. Changes in LIS dimensions observed with osmotic and/or hydrostatic pressure gradients are believed to reflect capacitative changes as opposed to resistive changes. convective water absorption or secretion may induce solvent drag of solutes from the lumenal or subepithelial compartments. cm2/h) for a solute through the transcellular (cell) or paracellular (paracell) pathways are incorporated in a diffusional permeability term of the form (DK/z). C. As discussed previously. binding the epithelial cells together near their apical (lumenal) domains. the following equations describe the contributions of cellular and paracellular flux: Jcell = Dceu" Kcell . an additional driving force must be incorporated in the intestinal model to provide for the influence of convective water flux across the paracellular pathway. mol/cm 3) and the total transporting area (A.h . Changes in the geometry of the space between epithelial cells (lateral intercellular space. K is a lumped partition coefficient representing both plasma membranes and possible aqueous/membranous partitioning in the cytosol. consisting of a permeability term (cm/h). where K represents the partition coefficient and the length of the relevant diffusional pathway. The significance of convective water flow across the paracellular pathway lies in the capacity of such flow to carry dissolved solutes in the convective stream.

As a second example. the diffusion coefficient for water. 2 is also dependent on D paracell. 1 and 2. as previously described for the diffusional fluxes. The significance of this dimensionless ratio can be appreciated by use of two limiting examples. A resulting value of zero for the diffusion coefficient ratio is associated with complete sieving of the solute from the convective water stream with no convection-dependent mass transport. The implication of such sieving or filtering as a result of convective flow. 2. The diffusion coefficient for solutes in the aqueous pathway may. This is to say that bulk water transport would move such a solute without any sieving. Consequently. For the sake of establishing common ground for discussion. For the sake of parallel arguments in Eqns. is that for a sufficiently soluble compound.346 H. let the solute diffusion coefficient be identical to that of water. the conventional use of the solute reflection coefficient [5] has been replaced by the ratio of the solute diffusion coefficient through the paracellular pathway with respect to the diffusion coefficient of water through the same pathway [33].N. filtering or reflection. the diffusional driving force through the cellular pathway may be enhanced. NELLANS recognize the paracellular pathway as a significant parallel route for intestinal mass transfer. These dimensions suggest that molecules slightly in excess of 3-5 kDa may traverse this pathway. The first case is characterized by the absence of a detectable diffusion coefficient for a solute of interest. no solute would be expected to traverse the paracellular pathway by either mechanism. Discussion of the active modulation of paracellular permeability will be found in a subsequent section. The physical dimensions of the paracellular pathway have been discussed previously and will no doubt be subject to continuing investigations. It should be added that the concentration on the opposite side of the paracellular pathway could be simultaneously decreased through wash-out. the solute entrained in the convective water stream across the intestine would enter the receiving compartment at the concentration in the originating compartment. The effect of this sieving would be to enhance transcellular diffusional net flux independent of mass transfer via the convective stream of water flow. the equilavent pore diameter for this pathway appears to lie between 10 and 30-50 A. It is not difficult to imagine that most solutes of interest as therapeutic agents would possess paracellular diffusion coefficients between the extremes of zero and that of water. Because the diffusional permeability t e r m (oParacell/XParacell)of Eqn. The form of the solvent drag or convective component of paracellular flux is dependent on concentration (C) and area (A). . approach or in unusual circumstances exceed. Such evidence exists for both animal [18-20] and human [17] investigations and recent studies have implicated a regulatory mechanism for changing the dimensions of this pathway [5]. but the sieving characteristics of the paracellular pathway must also be incorporated in this component of Eqn. however. most convectively driven solutes would be filtered to varying degrees by the paracellular tight junction and may even accumulate at higher concentration than in the bulk medium. In this instance.

That is to say that the paracellular pathway represents only 0.1% from functional transport studies in the rat.-. The cellular partition coefficient (K) represents another significant factor which can dramatically alter the balance of solute flux between cell and tight junction. The contribution of relative cross-sectional areas (Aparacen/Acen) for transport represents a significant difference between the two pathways. although it is conceivable that the cellular diffusion route may be significantly longer than the paracellular route.cell mparace. This has.1 and the reciprocal partition coefficient ranges from 1 to 10 -6. For lipophilic compounds (K.2 and 10 +4.l {/Jr Y. J~aracei. 1).10-2).paracel. With inclusions of area amplification of epithelial cells due to microvilli and limited diffusional area through the tight junctions.-~ 104). This is to say that for hydrophilic compounds (K. what is the significance of this pathway with respect to absorption and secretion relative to the cellular pathway? One approach to this question is to divide the expression for the paracellular flux (Eqn. For the sake of simplicity.1% of the available intestinal epithelial transport cross-section. in fact.PARACELLULAR INTESTINAL TRANSPORT: MODULATION OF ABSORPTION 347 11. The ratio of diffusion coefficients between paracellular and cellular pathways Dparacell/D call is likely to be greater than 1 in most cases. 1%. + 1) • l Dccll ~paracell "'4ccll "k. then the area of the paracellular pathway would be approximately 8% of that of a single cell of 5 #m diameter.5 kDa). Relative transport contributions of transcellular and paracellular pathways If the dimensions of the paracellular pathway are large enough to accommodate compounds with molecular radius approaching 15 A ( ~ 3. The contributions of the area and cell partition terms weigh heavily in favoring the transcellular pathway for transport of many compounds considered for oral delivery.1 #m. the area and partition coefficient product would predict that roughly equal mass transfer would occur across the two parallel pathways. then the product of the area ratio 0. the ratio of paracellular-tocellular flux can be analyzed with respect to ratios of several contributing parameters. Jcell Dparacell ]. If it is assumed that the majority of compounds of therapeutic interest possess partition coefficients which lie between 10 . a more reasonable ratio for the two pathways would be closer to 0. the paracellular pathway may contribute an equivalent fraction of transported mass when compared to the transcellular pathway. . Pappenheimer and Reiss [5] have calculated this cross-sectional ratio to be < 0. we will assume the ratio of pathway lengths (XCeU/gparacen)is approximately 1. 2) by the expression for cellular flux (Eqn. been confirmed for water transport in the rat [5]. OH20 / R (3) After simple rearrangement and simplification. since free solution viscosity is significantly less than that of the cellular plasma membranes. to this point of the discussion. no more than 1 in 106 of the transported molecules would be expected to traverse the paracellular pathway. At best. If one assumes that the width of the tight junctional complex is 0.2.

Acell • Dcell " K Zcell \ + Ap aracell . The significance of this back flux is that the net flux becomes the difference between the forward or absorptive. Convective flow and paracellular flux The contributions of convective water flux. particularly in vivo where the variables driving the flux from subepithelial space to lumen are not well controlled and are often entirely unknown. The importance of this observation is that net flux can be changed appreciably by changes in either the secretory or absorptive unidirectional flux. little or no consideration is given to the magnitude of the flux from tissue to lumen. As a result. 1 and 2 to yield an expression for net flux which provides one novel insight with respect to the influence of convective water flow. This is a very feasible value for intestinal water transport under physiological conditions where values for absorption can frequently approach 20 #1 × 10 -5 cm2/s [34]. the contribution of the paracellular pathway as a transport route increases. the net flux or difference of the two unidirectional fluxes is frequently measured simply as the rate of appearance of solute in portal blood or often as the disappearance of lumenal solute. has focused explicitly on unidirectional transfer of both solutes and water.348 H. as represented by Eqns. The discussion of increasing absorptive paracellular water flux as a mode of enhancing paracellular solute flux will be discussed in the section dealing with active regulation of paracellular dimensions. paracellular convective flow of 20 ktl-l-cm-2.2.4. Eqn. NELLANS 11. Jnet + ( . At non-zero values of Jv. H. For a value of unity for Jv/DH~o. areas (A) and the concentration difference . The value of the free diffusion coefficient for water at 35°C is approximately 3. In these measurements. normalized to unit path length. Op aracell x Zparacell ms sin / o (Cm Cs) (4) mparacell" Dparacell ' (Jv OH20 • C m . to intestinal transport are capable of significantly enhancing paracellular mass transfer. h-1 could increase the ratio of Jv/DH2o to 10 and dramatically enhance the relative contribution of paracellular flux. Net intestinal transport as a difference in unidirectional fluxes The discussion of intestinal transport via diffusive and convective pathways.Jv • Cs) Note that the first two bracketed terms simply describe the diffusive net flux in terms of permeabilities (DK/x). (Jv/DH2o) + 1. h . 1 and 2. The unidirectional fluxes are often difficult to measure empirically. the volume flow of water required is approximately 2 / d . the dimensionless ratio between Jv and DH~o is zero and the area ratio and cellular partition coefficient dominate balance between paracellular and cellular flux. h-1 Consequently. 4 combines the terms of Eqns.3.r. flux and the back or secretory flux.1.N. In the absence of convective water flux. c m .

the simple difference in concentration becomes the driving force for the net flux. the inclusion of a component in Eqn. . while pressure originating internally is readily . Owing to the fact that diffusional fluxes are most frequently governed by entropic forces. such as inadequate serosal vascular perfusion [35] or where epithelial permeation is not rate limiting. The result of this phenomenon is that the net convective flux of solute is essentially a unidirectional flux as contrasted to the net diffusional flux which is the difference between what may be large unidirectional fluxes. contrary to diffusive movement.) between the mucosal and serosal bathing media. Under conditions.5.) is frequently quite small compared to the musosal concentration (C.PARACELLULAR INTESTINAL TRANSPORT: MODULATION OF ABSORPTION 349 (C. resistance to flow drops and flow increases markedly toward the lumen [1820. when hydrostatic pressure is applied from the serosal surface. although not well understood on mechanistic grounds. Hydrostatic pressure differences directed from lumen to serosa induce a contraction of the lateral intercellular space and increased resistance to volume flow [18-201. . the mucosal concentration is the primary driving force for net diffusive flux. These observations on diffusive flux are well-accepted principles of mass transfer. The significance of this net unidirectional flow for solvent drag (convective) flux is that the intestine displays distinct rectification properties with respect to hydrostatic pressure differences. does transfer significant energy to oppositely directed streams. the cellular spaces expand.). may increase to such an extent that net diffusive absorption is substantially reduced.C. Implicit in this characterization is the fact that diffusional fluxes in opposite directions through the same pathway are essentially independent of one another.JVSm C. The contribution of convective water flow to solute transport occurs as a result of solvent drag of dissolved solute molecules. RectiJied convective water flow: physiological barrier to absorption Rectification of epithelial resistance to convective flow. Namely. pressure from exterior to the organism (lumen) does not efficiently drive water into the organism. the concentration in the serosal or subepithelial space (C. C. This is to say that no significant energy transfer occurs between solute or water molecules diffusing in opposite directions across the intestine. The practical implication of this principle is that for solutes which are transportlimited by their epithelial permeation. 4 based on volume or convective flow represents the novel contribution to the net flux expression. The random dissipation of the concentration gradient determines the net flux. Convective water flow. does have appeal on the teleological level. 11.). If we assume for the moment that paracellular cross-sectional area (Aparacelt) and solute sieving are invariant. then the controlling factor for convective transport lies (Drarace~~) in the difference of the products of unidirectional volume flow and source compartment concentration (FC. Consequently. This rectified behavior of the intestinal epithelium to pressure-driven convective flow has significant implications for net intestinal solute absorption.931. However.

One interesting observation in this regard is the continuing difficulty in the pharmaceutical industry associated with the oral delivery of putative renin inhibitors. The efficiency of such a scheme in the control of internal water homeostasis is both simple and elegant.350 H. The rise in interstitial pressure could initiate either a decrease in net water absorption or potentially. Interstitial pressure is controlled by capillary hydrostatic pressure leading to lymphatic drainage and intestinal secretion. These studies demonstrated that as smooth muscle activity increases. 2). water secretion. The mechanism for the AII effect has not yet been elucidated. Recently. but hydrostatic pressure is the dominant force in controlling water secretion.26] have shown that in cat small intestine. Similar hemodynamic changes may be occurring in the small intestine such that AII stimulates water absorption and AII antagonism may lead to decreased absorption or frank secretion. what are the factors modulating water flow from tissue space to lumen and what effect do such flows have on net solute absorption from the lumen? The principle driving force for water flow from subepithelial tissue space to intestinal lumen is hydrostatic pressure.37].N. An additional source of control of interstitial hydrostatic pressure in the intestine emanates from intestinal smooth muscle. cellular and paracellular pathways of . the rate of water absorption declines and finally reverses to net secretion. Levens and Suvannapura [25. Testing of such a hypothesis requires sensitive measurements of hydrostatic pressure changes as well as identification of the sites of AII vasoconstriction. Very small osmotic pressure gradients within the lateral intercellular space are responsible for water absorption. such as Vibrio cholerae and Escherichia coli [39. Such a constriction decreases hydrostatic pressure and increases osmotic pressure in downstream capillaries resulting in increased water reabsorption from the proximal tubule lumen [24]. In the latter cases. Although the magnitude of intestinal interstitial pressure is extremely difficult to measure accurately [36. that hydrostatic pressure within the intestinal tissue mass could be elevated during waves of either spontaneous contractions in the fed state or during migrating myoelectric complexes in the fasted state. but a striking parallel surfaces with respect to effects of AII in the kidney proximal tubule where this peptide exerts sensitive control over water balance through constriction of efferent glomerular capillary sphincters. Is it possible that these highly potent compounds may lead to local reductions in AII levels in the intestinal vasculature which in turn blunts further absorption due to an increase in interstitial hydrostatic pressure? Such a hydrostatic pressure increase could be envisioned to carry absorbed solute in the lateral interstitial spaces back to the intestinal lumen in a cycle of futile turnover with little net absorption (Fig.40]. If lumenal pressure is of relatively little significance in the control of intestinal water flow. vasoactive angiotensin II (AII) is a significant factor controlling absorption of water. Similar observations have been made in response to toxigenic bacteria. One can envision. a striking correlation between duodenal smooth muscle activity and duodenal water transport was reported in humans [38]. N E L L A N S dissipated through water flow to the outside.

When oral bioavailability was assessed. While far from definitive. Reductionof absorption by paracellular water secretion. The common mechanism for these effects is. conceivably. The mechanism for such an effect would be by recycling diffusionally absorbed solutes from the lateral intercellular space back to the lumen before such solutes can gain access to lymphatics or mucosal capillaries. Another example of a causal link between motility and absorption lies in unpublished observations from this laboratory with derivatives of erythromycin. These compounds are documented to possess a broad range of motilityinducing properties in the dog. Hydrostaticpressure in this spaceregulatesintestinalsecretionas well as lymphaticand capillaryabsorption. intestinal motility may change interstitial hydrostatic pressure such that absorption of lumenal solutes is either slowed or in some cases halted. as illustrated in Fig. 2. The above findings suggest that net flux of solute transport by the intestine may possibly be influenced by paracellular convective water transport. such a correlation suggests that similar to the human studies above [38]. a striking inverse correlation was observed between motility induction potency and systemic bioavailability. . Recyclingof absorbed solutes and wash-out of diffusionalfluxesdiminishesabsorption. The influenceof secretorywater flow betweenadjacentepithelialcellsis depictedto includea reductionin net absorption througha solventdrag effect on solutes(X)in the lateral intercellular space. 2. an increase in Intestinal Epithelial Cell Lumen _ ~ ~ Lymphatics ~ Capillaries Fig. These effects stem from influences of the absorbed solutes on water flow from intestinal tissue to lumen.PARACELLULARINTESTINALTRANSPORT: MODULATION OF ABSORPTION 351 secretion are stimulated making the identification of the paracellular secretory component more difficult. The absorption of solutes by entirely predictable diffusional routes may be thwarted by physiologically based mechanisms.

N E L L A N S the tissue or interstitial hydrostatic pressure. Studies by Meza and associates [54] in kidney epithelial cells have also demonstrated a specific effect of the cytoskeletal perturbing activity of cytochalasins on tight junctions. resulting in increased water and solute flow.52] and c-AMP [53] may also control permeation of solutes through the paracellular pathway of cultured epithelial cells have all added to the evidence in favor of a modulated barrier function for the tight junction. This convective effect can wash out solutes deposited in the tissue space as a result of absorptive diffusion through the cells.352 H. since it is not currently known how selective absorption can be controlled by such a process. Similar findings have also been reported for intact intestinal epithelia [46. Consequently. III. however.48]. this effect may manifest itself as a dramatic rise in paracellular solvent drag. such as glucose and amino acids.1. The following discussion will identify. Observations that protein kinase C [49]. Discrimination among solutes absorbed by such changes is both intriguing and perplexing. Investigations from Madara's laboratory [32.N. Other laboratories have advanced similar hypotheses [47. In addition to such passive reactions to physical forces.46] have suggested that the cytoskeleton may control the physical geometry of the tight junction through contraction of actin microfilaments. [56. Further. From the extensive structural studies of Madara. These changes in electrical conductance are paralleled by increased fluxes of sodium and the specific paracellular probe. independent of partitioning properties. Madara and coworkers [46] have also been able to demonstrate that perturbations of the tight junctions of cultured human colonic epithelial cells by cytochalasin D lead to decreases in electrical resistance. Active control of paracellular absorption III. Mechanism of change in intestinal permeability Secretory water flow and associated mechanisms giving rise to increases in pressure in the subepithelial space are associated with passive responses of the paracellular pathway. suggesting that paracellular pathway permeability may be increased several-fold in response to nutrients. intestinally absorbed solutes can be restricted from access to systemic compartments by local recycling from epithelial tissue space to intestinal lumen. The active components of these structurally based changes are the actin microfilaments of the tight junctional complex. as well as the functional studies of Madara and Pappenheimer and their coworkers. calcium [50. the paracellular pathway. may have the capacity to alter actively its resistance to solute and water transport. cellularly controlled changes in paracellular permeability as well as functional transport results correlated with such changes. particularly the tight junction. both the structural basis for such active. Stevenson et al. mannitol.57] and D'Angelo-Siliciano and Goodenough [61] have .55] by Madara and collaborators. a significant body of literature has emerged. When coupled to the propensity of the small intestine to lower resistance and thereby increase flow when subjected to pressure from the serosal side.

depending on the direction of the gradient. the number of strands did not decrease and the most apical strand never displayed discontinuities despite significant increases in HRP translocation. believed to contribute mechanical integrity to the tight junctional complex [63].60]. that osmotic loads may alter tight junction structure [59]. reports that strand density and depth increase with cell age and migration up the villus in guinea pig cecum. The secretory crypt cells may benefit more from such increased junctional leakiness owing to their function as a source of net salt and water movement into the lumen [67].PARACELLULAR INTESTINAL TRANSPORT: MODULATION OF ABSORPTION 353 recently isolated a protein (ZO-1) from intestinal epithelium. it is undetermined whether a spontaneous tip-to-crypt villar gradient of tight junctional leakiness may exist. these 'passive' effects may also be under the control of the cellular cytoskeletal system. but this observation could well be linked to physical factors such as osmotic pressure rather than a specific effect on a cellular control system through the cytoskeleton. Schulzke and coworkers [64] have reported that the density and depths of tight junctional strands decreases from crypt to villus in rat jejunum and that in experimentally induced blind loops. that. presumably to maintain solute gradients. His morphological studies revealed that while. the resultant void was filled simultaneously. He concluded that voids in the continuous villar tip epithelium were so transient that such a pathway could not account for the magnitude of paracellular flux observed with intact tissue. It is of interest to speculate. Marcial and Madara [62] have suggested that the cells at the tips of intestinal villi may be more vulnerable to changes in junctional structure than cells of the crypts. Further. the density and depth of strands increased at all sites. These studies. Pappenheimer and Reiss [5] have been able to demonstrate in rat small . Madara [66] in a subsequent investigation explored the possibility that the villus tip may be the most likely site for significant paracellular flux as the result of shedding of mature cells. which may represent the junctional complex linking cytoskeleton with tight junction. in guinea pig small intestine. however. Madara's group has also been able to demonstrate. It remains to be determined if cellular signal transduction in addition to simple mechanical stress is requisite for induction of rectified responses in both water and solute flux. Absorption efficiency would be significantly limited by increased leakiness. decreased their crosslinking. which employed osmotic gradients to induce changes in horseradish peroxidase (HRP) translocation. villar tip cells did exfoliate.51]. but the villus tips have also been implicated as the primary sites of small intestinal water absorption. Mora-Golindo [65]. The charge selectivity of the paracellular pathway appears to favor cations [2. on the other hand. since the effects of hydrostatic and osmotic pressure exhibit distinct properties of rectification [1820. also revealed that the junctional strands. indeed. Additional inquiry is required before meaningful conclusions can be drawn with respect to variations in paracellular conductance as a function of a tip-tocrypt villar gradient. As a consequence of the above studies. The villar tips may be more susceptible to decreases in tight junctional integrity.

consequently. through a link to the tight junction. as well as the possibility of additional control signals. most probably serves as the mechanism of action to explain changes in the geometry and. Consequently. as suggested by Madara and Pappenheimer [32] and Atisook et al. gall bladder [95]. It is quite likely that cellular responses may remain intact following isolation. Other epithelia and in vitro/in vivo comparisons Additional studies are necessary to elucidate the signal transduction system between extracellular control factors and cellular mediated changes in paracellular permeability. determine the significance of changes observed in isolation. effects on absorption may be greatly overestimated. Gustke et al.3. Permeability changes and in vivo absorption If for the moment we assume that the cytoskeleton is responsible for changes in tight junctional geometry and permeability. including proximal kidney tubule [21-23]. the final arbiter of physiological significance is the intact animal where nuance of driving force. cornea [99] and pancreas [101]. however. an anionic solute may be completely impermeant through the paracellular pathway. The cytoskeleton. tight junctional permeability may increase in the presence of luminal glucose. but the balance of forces determining net water and solute fluxes may be dramatically altered by the in vitro conditions. in extrapolating conclusions drawn from studies on isolated tissues or cultured cell monolayers to the in vivo state. Caution must be exercised. the permeability of the intestinal paracellular pathway. For example. [71]. 111. but if interstitial hydrostatic pressure is decreased due to experimentally induced decreases in capillary blood pressure. forces and resistances for the paracellular pathway have not yet been accounted for in the in vitro or in situ studies reported to date. that with increased interstitial hydrostatic pressure in vivo. cortical collecting duct [70]. while the specific elements of paracellular permeability may be studied in detail with isolated or cultured systems. with four negative charges. III.2. Other investigators [68] have published similar experiences. in the example above. Many other epithelia have been characterized with respect to plasticity of tight junctional conductance and permeability. Several attempts from this laboratory (unpublished results) to stimulate paracellular solute flux in intact rats using lumenal glucose have not met with success.354 H. behaved as an ideal osmotic solute. they could not detect any significant intestinal transport despite a relatively small molecular mass (175 Da). These investigators reported that the ferrocyanide ion.N. higher permeability may be offset by secretory water flow such that little or no net increase in absorptive flux is observed. what is the magnitude of such changes and what size solutes are likely to cross this barrier? Pappenheimer and . It is conceivable. NELLANS intestine that with a charge to mass ratio high enough. [58] have investigated cation selectivity for human ileum and jejunum and report that up to 55% of total conductance is attributable to the cation selective paracellular pathway of jejunum. suggesting that the full complement of flows.

suggesting transport by convective flow (solvent drag). accounting for 50% of total water absorption. In these studies. These structural studies were carried out. lead to not only increases in water absorption but also an increase in the permeability of the tight junction.5 kDa) and PEG (4 kDa) with only slight restriction.P A R A C E L L U L A R INTESTINAL TRANSPORT: M O D U L A T I O N OF ABSORPTION 355 Reiss [5] have reported an intriguing series of studies on paracellular transport using anesthetized rats. . Further. The absorption of creatinine. Functional electrical impedance measurements [69] as well as tight junctional morphology studies Intestinal Epithelial Cell Lumen Lymphatics I-I20 Capillaries Fig. Since glucose absorption was also observed to be a linear function of water absorption. The cross-section of the tight junction (broken lines) has been postulated to increase in the presence of glucose and amino acids as discussed in the text. not in rat. Pappenheimer and Reiss postulated that active sodium transport. the dimensions of small intestinal paracellular tight junctions were calculated at 100 A diameter. 3. but in isolated segments of hamster small intestine or in anesthetized hamster in situ. lumenal glucose (23 mM) doubled net water absorption and the increase in the flux of the three hydrophilic solutes doubled as well. indeed. Independent investigations by Madara and Pappenheimer [32] demonstrate that glucose and amino acids. Such an increase is suggested to be associated with an increase in intestinal absorption via both diffusion and convection (solvent drag) when lymphatic and capillary drainage allow adequate removal of water and solutes. 3. as illustrated in Fig. passing molecules the size of inulin (5. generates the requisite driving force for water absorption through the paracellular pathway. PEG and inulin were all demonstrated to be linear functions of water absorption in the small intestine. Enhanced absorption via paracellular diffusion and convection. stimulated by the addition of glucose.

lead to both significant decreases or increases in integrated permeation such that in vitro . consequently. increased permeability. One possible explanation for such results may lie in lower secretory convective flow in anesthetized rats resulting from lower mucosal capillary perfusion pressure. It will be necessary to critically evaluate intact animal models to ascertain whether the absorption promoting effects of glucose and amino acids observed in vitro or in situ can be translated to comparable enhancement in uncompromised animals. consequently. suggesting that the uncompromised animal may not respond as dramatically to glucose stimulation as the anesthetized animal. Such studies. Where permeation through the paracellular pathway is a significant fraction of total flux. The influence of additional in vivo factors. provide consistent permeability data allowing rank ordering of compounds. the magnitude of the results was appreciably lower than the prediction of the anesthetized model. While this observation is qualitatively supportive of the glucose-induced permeability effect seen in anesthetized rats. integrated permeation of solute from lumen to blood.5-fold more exogenously administered oral creatinine than controls consuming glucose-free water. active modulation of tight junctional permeation and the more traditional role of capillary blood flow clearing solute from the subepithelial space [35]. in fact. Lower secretory water flow may cause less recycling of absorbed solute and. Certainly. lending additional credence to the physiological significance of these changes. greater net absorption. alanine and leucine may be responsible for initiating contraction of the junctional actomyosin leading to expanded geometry of the occluding junctions and. the in vitro model may bear little or no relation to the in vivo state. the rates of glucose-stimulated absorption of creatinine in the anesthetized animals were nearly threefold greater than in glucose-free controls. as recently exemplified by investigations of Gross and Sweetana [81]. Pappenheimer [69] reports that his survey of the literature indicates glucose-dependent changes in electrical impedance of rat small intestine are seldom observed in isolated sheets and may be limited to tissues that are oxygenated comparably to in vivo conditions. These dynamic factors may. the design of the unanesthetized animal study may not optimize the absorption promoting potential of glucose. be explained as a result of paracellular recycling. Passive permeability characteristics of intestinal epithelium are often measured in vitro with flat sections of the intestinal mucosa stripped of underlying musculature. such as solvent drag.N. can all play significant roles in the dynamic. Indeed. However. NELLANS [32] support the hypothesis that glucose. Decreased net creatinine absorption in t h e unanesthetized rat could. Further. consequently. however.356 H. The critical dependence of the permeability (impedance) change on adequate oxidative metabolism also suggests that significant differences may exist between anesthetized and unanesthetized rat models. The increased electrical impedance observed with glucose and amino acids was both oxygen dependent and reversible. Pappenheimer and Reiss [5] report that rats given access to 20% glucose in water excrete approximately 1.

Further. human colonic epithelial cell line. The paracellular pathway in disease states Changes in permeability of the intestine have been attributed to specific alterations in the paracellular pathway associated with a variety of pathological conditions. Hecht et al. Model systems exist for in vitro investigation.73. Another intriguing example of paracellular plasticity has been described in the studies of McRoberts and coworkers [72]. virtually nothing is known about methods to prevent such disease-induced functional degradation of the intestinal epithelial barrier. significantly lower paracellular permeability of the colon will require development of cultured small intestinal cell lines before tissue specific data are available from these in vitro models. Insulin is reported to be responsible for a significant reversible increase in . Fleisher and coworkers [68] have attempted to capitalize on the absorption enhancing potential of glucose. In these studies. [44.P A R A C E L L U L A R INTESTINAL TRANSPORT: M O D U L A T I O N OF ABSORPTION 357 studies must always be compared to the observed in vivo results to confirm utility of a selected in vitro technique. [75] have suggested that both the small and large intestine demonstrate increases in paracellular pathway permeability in Crohn's and possibly celiac disease. Recently. Madara and Trier [79] note. It is not unlikely that the ability to break the barrier between internal and external (lumenal) environments is a key factor in the progress of these disease states. as well. given significant lumen to blood concentration gradients. [76] and in mice. IV. bacterial toxin A is responsible for a massive loss of mucosal integrity due to disruption of the tight junctional complexes as demonstrated in both the human colonic T-84 epithelial cell line. as demonstrated by Madara et al. Hollander et al. but a satisfactory cell culture system for small intestine has not been developed to date. employing phenytoin as the solute targeted for solvent drag-promoted absorption. At the current time. [78]. a significant disruption of tight junctional complexes of both villus tip and crypt cells in celiac spruce. These results suggest that other driving forces may be influencing net intestinal absorption in addition to the elegantly characterized component of absorptive solvent drag. Heyman et al. Madara and their coworkers. These observations of decreases in intestinal mucosal restriction of lumenal molecules from subepithelial tissues suggest that breaching the epithelial barrier could be a significant factor in the etiology of these conditions. The well-known Caco-2 and T-84 colonic cell lines have certainly provided initial insight to transport pathways of the colon and perhaps the small intestine. [77]. With the infection of Clostridium difficile. only marginal improvement in absorption was noted in glucose-treated rats. described by Pappenheimer. inflammation and leukocyte transmigration across T-84 monolayers have also been associated with reversible decreases in electrical resistance and increased fluxes of paracellular probes consistent with increased tight-junctional permeability. These investigators have recently demonstrated a unique effect of insulin in the T84. However.74] and Dawson et al.

358 H. the tight junctional complex is frequently perturbed in animal studies with agents that chelate calcium ion. however. such as EDTA. While these agents may be affecting an increase in junctional permeation by inhibiting perijunctional actomyosin [31]. as may be the case for sugars and amino acids are possible. It is a more desirable candidate as a paracellular probe for peptidic drugs than more conventional hydrophilic molecules. The implications of hormonal control of paracellular permeability add an additional dimension to the dynamic control of this route of absorption. alcohol [85] and other irreversible procedures are not likely to be viable alternatives for clinical utility in drug delivery. the most probable route for the observed intestinal translocation of the compound.82-84]. Recently. EGTA [14. Bioavailability of PEG-898 monomer has been reported [102] to be in the 5-10% range in man. including enterotoxins or inflammatory peptides as in the cases above.44. have investigated the characteristics of PEG transport in the rat intestine. Paraceilular probes The identification of an appropriate probe molecule(s) for the paracellular pathway is of great utility in assessing the site of action of agents which may modulate intestinal permeability. although an argument could be made for increased efficiency of glucose absorption under conditions of restricted dietary intake. the effects are frequently irreversible. NELLANS tight-junctional permeability generated over a period of 3-4 days. This result suggests that peptides in this size range. with appropriate formulation and water flow conditions. Hollander and his collaborators. V.73]. owing to greater molecular size. PEG-900 was shown to be nearly completely excluded from entry across isolated brush-border membranes of rabbit intestinal epithelial cells. such as mannitol. might be expected to approach bioavailabilities as high as 20-30%.N. In an effort to increase the paracellular route of drug delivery. calcium chelation [31] and some bile salts [80. This irreversible degradation of tight junctional integrity may lead to indiscriminate penetration of the epithelium by lumenal solutes. Reversible modes of increasing junctional permeability. PEG-900 represents a spectrum of polymers with molecular weights from 200 to 1100 and hydrophilic properties favoring paracellular translocation.15] and citrate. With an octanol/water partition coefficient of 7. looking for a probe to assess changes in human intestinal permeability [43.9 • 10 4. The Hollander group found that PEG-900 behaved as if the only pathway for translocation was identical to that available for bulk water . The aqueous environment of the paracellular pathway is. PEG-900. The physiological implications of such a long time-course for an insulin effect in the intestine are not readily apparent. may serve such a function. Hollander and his coworkers [42] have provided compelling evidence that poly(ethylene glycol). consequently. Such hormonal regulation is a possibility which calls for additional study of the active cellular mechanism for permeability modulation.

Further. if net water reabsorption is not required by the organism. Results from our own laboratory.26].P A R A C E L L U L A R INTESTINAL TRANSPORT: M O D U L A T I O N O F ABSORPTION 359 flow. have confirmed the utility of PEG as a paracellular probe and the synthesis of tritium-labeled polymers of known molecular weight [45] has been helpful in determining the size discrimination of the paracellular pathway both in vitro and in vivo. volume depletion and osmotic activity with aldosterone and angiotensin II in control of whole animal water homeostasis. Conclusion Intestinal absorption through the aqueous environment of the paracellular pathway represents an unexploited route for the absorption of molecules whose weights may approach 5 kDa. Consequently. it appears that the intestine via the reninangiotensin system participates. These investigators conclude that PEG-900. both conditions which reduce net water absorption. aldosterone is known to have water conserving effects on the colon [100]. Because of the myriad interactions of sodium loading. in parallel with the kidney. In this case. to techniques which may stimulate solute absorption via the paracellular route. the osmotic activity of the extracellular fluid may determine the efficacy of stimulation of intestinal fluid (and solute) absorption by glucose and amino acids. VII. Stimulation of water absorption increased PEG absorption. in the homeostatic control of water balance. They reported reduced absorption with increased lumenal osmolarity and increased lumenal hydrostatic pressure. factors such as sodium loading and/or volume depletion may result in increased intestinal solute absorption through the actions of angiotensin II and aldosterone. by virtue of both its size and low partition coefficient. As another example. many combinations of factors can be explored in regulating intestinal drug absorption. as well as several others [86-92]. suggesting that colonic drug delivery might also be modulated by convective water flux. is a viable probe for paracellular permeability and would be of utility in both clinical and animal studies for monitoring changes in paracellular permeability. with editorial license. With equivalent pore diameters as large as 30 to 100 A. this pathway provides a portal of significant specific cross-section for . VI. From the observations of Levens [25. Exploratory studies The argument that secretory convective water flux may decrease absorption of lumenal solutes may be extended. water recycling from lateral space to lumen may occur as a result of the renin-angiotensin control of mucosal capillary uptake. The implication for such a control mechanism is that factors favoring water conservation (increased absorption and/or decreased secretion) may be used to test the hypothesis that convective water flow is a dominant factor in controlling both the direction and magnitude of paracellular drug transport. Such exploratory studies may well identify a drug delivery niche for exploiting the aqueous intestinal paracellular pathway.

Drug Delivery to the Gastrointestinal Tract. 863-868. 107. S. 16. and Reiss. Meza. (1990) Intestinal calcium transport: interplay of paracellular and cellular pathways. Whether the paracellular junctional resistance will prove to be a portal for oral delivery. The paracellular pathway is capable of both increasing and decreasing intestinal absorption.. Wilson (Eds. M.. New York. R. Pharmacol. J. M. J... 5 Pappenheimer. Frizzell. Biol. Hardy. References 1 Collander. 31-61. K.G. and Cereijido. 43 56.. Membr. 123 136.1%. remains to be established. Lynch. G.L. J. Rev. which can be actively controlled in a reversible fashion. A. S. G. D. J. Tox.G. Physiol. (1981) Occluding junctions in cultured epithelial monolayers. NELLANS absorption of high-potency therapeutic compounds. Membr. stimulating and in some cases inhibiting. G. (1985) Absorption potential: estimating the fraction absorbed for orally administered compounds. and Amidon. (1972) Ionic conductances of extracellular shunt pathway in rabbit ileum. 318 346. Amidon. 2 Frizzell. 11.A.360 H. Bot.A. 59. The possibilities are beginning to be explored and dogma that intestinal drug permeation is dominated exclusively by high partition coefficients and absence of charge must be challenged.D. I. (1981) Barrier function in epithelia. LS~zaro. R. While the total area of the paracellular pathway relative to the cellular pathway may approach 0. (1974) Ion transport by mammalian small intestine. 241.N. 4 Nellans. J. the challenge of the future is to harness its plasticity to selectively and reversibly enhance oral bioavailability of highly potent peptides. (1990) Absorption of poly(ethylene glycol)s 600 through 2000: the molecular weight dependence of gastrointestinal and nasal absorption. Biol.N. Physiol. 10 Donavan. 9 Taylor. B. The importance of including the physiological effects of orally delivered drugs on hydrostatic pressure has been discussed in the context of divergences between in vitro and in vivo observations. 3 Schultz. (1933) Permeabilitatsstudien an Chara ceratophylla. Chvez de Ramirez. Res. J. Am. Flynn. G275-G288. The more integrated approach must include an aqueous route of delivery between cells for potent compounds with significantly higher hydrophilic character.C. Pharm. In: J. A. Ellis Howard.L. 8 Dressman. Davis and C. 133-145. and Leahy.). 11 Gonzfi. 7 Lien. R. 51 92. The importance of hydrostatic pressure in the subepithelial space can dramatically alter net absorption.E. 6 Powell. (1987) Contribution of solvent drag through intercellular junctions to absorption of nutrients by small intestine of the rat. 100. C96-C102. E. Gen.S. D. H. 101-108. Ann. I. S. 12 Cereijido. such as peptides and peptidomimetics. and Nellans. (1981) Structure-activity relationships and drug disposition.. D. Acta. Sci. (1989) Establishment of tight junctions between cells from different animal species and different sealing capacities.N. and Schultz. 240. Physiol. and Martinez-Palomo. D. 36. 21.R.G. Mineral Electrolyte Metab. J. the influence of convective water flow provides a compensating factor in balancing the relative transport contribution of the two routes..B. delivery to the central systemic compartment. Annu. Rev. Am. pp. 588-590. M. .J. H. J. (1989) Models for intestinal permeability to drugs.lez-Mariscal. 74.L.Z. 7.G. 1-114. J. and Fleisher. Fem. Pharm. Physiol.

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