Transport permeability coefficient

The effect of the substituents on chain mobility and chain packing has been related to the gas transport properties [209]. Role of symmetry of methyl group placement on bisphenol rings in PES shows the permeability coefficients in the following order ... 

The gas sorption and transport properties also depend on the bisphenol connector groups [210]. The permeability coefficients for all gases rank in the order ... 

The equations used to calculate permeability coefficients depend on the design of the in vitro assay to measure the transport of molecules across membrane barriers. It is important to take into account factors such as pH conditions (e.g., pH gradients), buffer capacity, acceptor sink conditions (physical or chemical), any precipitate of the solute in the donor well, presence of cosolvent in the donor compartment, geometry of the compartments, stirring speeds, filter thickness, porosity, pore size, and tortuosity. 

Shah et al. [51] demonstrated the use of a donor-receptor compartment apparatus separated by a cell monolayer to estimate membrane transport parameters. Permeability coefficients, P, were calculated as... 

The effective permeability coefficient is composed of the permeability coefficients for the various transport barriers in series—the ABLs, the cell monolayer, and the filter support ... 

The plot of permeability coefficient versus molecular radius in Figure 10 shows the interdependence of molecular size and electric charge. The permeability of the solutes decreases with increasing size. The protonated amines permeate the pores faster than neutral solutes of comparable size, and the anions of weak acids permeate the pores at a slower rate. The transport behavior of the ionic permeants is consistent with a net negatively charged paracellular route. These results are phenomenologically identical to those found in the transport kinetics of... 

The quantitative role of electrical factors affecting the transport of charged molecules is obtained by comparing permeability coefficients with the permeability coefficient P(Lcdi for molecular size-restricted diffusion independent of the charge on the molecule (i.e., the neutral image). With Eqs. (45) and (46) one obtains... 

IV. FILTER SUPPORT TRANSPORT KINETICS A. Permeability Coefficient... 

The summary of Pe values for the steroids as a function of stirring rates is found in Table 11 and their correlations with log PC (n-octanol-water) in Figure 20. The transport kinetics of the relatively hydrophilic hydrocortisone and dexa-methasone are controlled by passive diffusion across the cell monolayer. On the other hand, the Pe values of testosterone and progesterone are highly dependent on stirring rate. The results for testosterone are used to obtain the relationships between the effective permeability coefficients of the ABL on the donor and receiver sides and the stirring rate, using the linear expression (see Eq. (69)]... 

The transport of both solute and solvent can be described by an alternative approach that is based on the laws of irreversible thermodynamics. The fundamental concepts and equations for biological systems were described by Kedem and Katchalsky [6] and those for artificial membranes by Ginsburg and Katchal-sky [7], In this approach the transport process is defined in terms of three phenomenological coefficients, namely, the filtration coefficient LP, the reflection coefficient o, and the solute permeability coefficient to. 

Here, we briefly describe the automated Caco-2 assay used at the research site in AstraZeneca R D Molndal. The solubility of the test compounds is measured (or theoretically predicted) before they are run in the Caco-2 assay. In order to be able to make correct determinations of the permeability coefficient, the substance must be dissolved when added to cell monolayer in the transport experiment. Compounds with insufficient solubility are therefore not tested. We generally apply a test concentration of 10 pM, but in specific projects or under certain circumstances a concentration of only 1 pM is applied. The test compounds are first prepared in DM SO solution (1 mM) on a parent plate and are then diluted in transport buffer to give a final drug concentration of 10 pM (solution containing 1% DMSO) when added to the cell monolayers. 

In our own studies to establish an in-house correlation between Caco-2 permeability and extent of drug absorption in humans, a set of 25 model drugs was used (Table 5.1). The importance of concentration and pH conditions were investigated and transport was studied both in apical to basolateral (absorptive) and basolateral to apical (secretory) directions. The apparent permeability coefficients were determined at concentrations of 10, 50, and 500 pM, and at two different settings of apical/basolateral pH values 6.5/7.4 and 7.4/7.4. The marker compounds represented a good diversity in molecular structure and transport properties and covered a range of low (<20%), moderate (20-80%) and high (>80%) extent of absorption in humans (Tab. 5.1). 

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. 

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). 

From an industrial perspective, quantitative knowledge of the existence of different transporters within the cellular system used in screening procedures is of major importance as it can influence both the predictive value of the permeability coefficients and interpretation of the results. In addition, information on species differences or similarities or discrepancies between cell culture models and animals now provide an important basis for the scaling of data during the early phases of drug discovery for animals or humans [48]. 

In Figure 2 we presented the permeability coefficient K of oxygen as a function of the mean gas pressure experimentally obtained for a sample of porous material from acetylene black modified with 35% PTFE. The experimental linear dependence is obtained. The intercept with the abscissa corresponds to the Knudsen term DiK. The value obtained is 2,89.1 O 2 cm2/s. The slope of the straight line is small, so that the ratio K,/ Dik at mean gas pressure 1 atm. is small ( 0.1) which means that the gas flow is predominantly achieved by Knudsen diffusion and the viscous flow is quite negligible. At normal conditions (1 atm, 25°C) the mean free path of the air molecules (X a 100 nm) is greater than the mean pore radii in the hydrophobic material (r 20 nm), so that the condition (X r) for the Knudsen-diffusion mechanism of gas transport is fulfilled. 

As noted earlier, the choice of species for experimentation is critical due to anatomical differences and it may also reflect species, as well as individual, differences in the expression/activity of transporter and metabolic proteins [43, 44], Since the fraction absorbed across buccal mucosa in vivo is not established for many compounds in different species including humans, the potential existence of a correlation between in vitro permeability coefficients in freshly isolated pig, dog, monkey, and human buccal mucosa was investigated (Figure 7.3). The correlation coefficient obtained for porcine and canine tissue was poor (0.65 and 0.67, respectively, at the 95% confidence level). Results for relatively high permeability compounds in porcine tissue resemble those previously reported where permeability coefficients were by an order of magnitude... 

MDCK Madin-Darby canine kidney (MDCK) cells have received attention as an alternative to Caco-2 cells for permeability measurements. When grown under standard culture conditions, MDCK cells develop tight junctions and form monolayers of polarized cells. The main advantage over Caco-2 cells is the shorter culture time to confluence (3-5 days). The transep-ithelial electrical resistance of MDCK cells is lower than that of Caco-2 cells and thus, closer to the TEER of the small intestine in vivo. The permeability coefficients of hydrophilic compounds are usually lower in Caco-2 cells than in MDCK cells, which is consistent with the lower TEER values for MDCK cell monolayers. The nonhuman (canine) and nonintestinal (renal) origin of MDCK cells is considered as a disadvantage. They have low expression levels of transporter proteins and low metabolic activity [34], MDCK cells that are stably transfected with P-gp/MDRl are often proposed as an alternative for Caco-2 cells to study bidirectional transport of compounds and, more... 

Either Transwell inserts or side-by-side diffusion chambers can be used for transport studies. Bode et al. have provided an excellent review on this subject [60], Briefly, cells are incubated for 30-60 min with a buffer solution. To initiate the transport study, a transport buffer containing the drug under investigation is added to either the apical or the basal chamber depending on the transport direction of interest. At predetermined time points, the respective receiver chamber is sampled and the withdrawn volume is replaced with the same volume of fresh buffer. The permeability coefficient (Papp) is calculated and the ratio of /apP in the basolateral-to-apical direction versus that in the apical-to-basolateral direction gives the efflux ratio. These sort of transport experiments are well suited to determine if drugs/xenobiotics are substrates of the placental efflux proteins. 

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