Effects of Digestion, Cell Culture Media, and Mucous on the Physical Properties, Cellular Effects, and Translocation of Polystyrene and Polymethacrylate Nanoparticles.

The discovery of plastic and metal nanoparticles in organisms, foods, and beverages has generated numerous studies on the effects of these particles on the barrier cells and their subsequent absorption into the body. Following ingestion, nanoparticles travel down the gastrointestinal tract (GIT), and their physicochemical characteristics change in response to the change in proteins and pH during their digestion. We measured the translocation of digested nanoparticles across a co-culture monolayer of Caco-2 and various combinations (1:9, 5:5, and 9:1) of HT29-MTX-E12. The in vitro model of the intestine was used to determine the translocation of digested 20 nm polymethacrylate (PMA) particles and the accompanying monolayer barrier effects after a 72 h exposure. The in vitro digestion increased the agglomeration and hydrodynamic diameters and decreased the surface charge of the nanoparticles. For NH2-functionalized polymethacrylate nanoparticles (PMA-NH2), the diameters increased from 57 nm (water) to 3800 nm (media), or 2660 nm (chyme). These nanoparticles compromised the integrity of the monolayer (trans-epithelial electrical resistance, Lucifer yellow translocation) and translocated across all the cell ratio configurations. Digestion can have a large effect on nanoparticle agglomeration and surface charge. Excess mucous was not seen as a barrier to the translocation of PMA-NH2.

Solid nanocrystalline dispersions of ziprasidone with enhanced bioavailability in the fasted state.

Reducing the absorption difference between fed and fasted states is an important goal in the development of pharmaceutical dosage forms. The goal of this work was to develop and characterize a solid nanocrystalline dispersion (SNCD) to improve the oral absorption of ziprasidone in the fasted state, thereby reducing the food effect observed for the commercial formulation. A solution of ziprasidone hydrochloride and the polymer hydroxypropyl methylcellulose acetate succinate (HPMCAS) was spray-dried to form a solid amorphous spray-dried dispersion (SDD), which was then exposed to a controlled temperature and relative humidity (RH) to yield the ziprasidone SNCD. The SNCD was characterized using powder X-ray diffraction, thermal analysis, microscopy, and in vitro dissolution testing. These tools indicate the SNCD consists of a high-energy crystalline form of ziprasidone in domains approximately 100 nm in diameter but with crystal grain sizes on the order of 20 nm. The SNCD was dosed orally in capsules to beagle dogs. Pharmacokinetic studies showed complete fasted-state absorption of ziprasidone, achieving the desired improvement in the fed/fasted ratio.

The use of gastrointestinal intubation studies for controlled release development.

AIMS: This review describes clinical results of gastrointestinal intubation studies of eight controlled release (CR) candidates under development during the 1990s and offers suggestions for determining why, when and how to conduct human intubation studies. METHODS: Experience with the administration of the following eight compounds to various regions of the gastrointestinal tract is described: CJ-13,610, CP-195,543, CP-331,684, CP-409,092, CP-424,391, azithromycin, sertraline, and trovafloxacin. Also included are human pharmacokinetic studies with prototype CR dosage forms for CJ-13,610 and CP-424,391. RESULTS: Intubation studies, while appearing invasive, are safe and not unpleasant procedures that have been found to be valuable in the development of CR formulations. CONCLUSIONS: The following recommendations are made regarding intubation studies: (i) no intubation study is recommended for compounds with high permeability, since these compounds are likely to be well absorbed from the colon; (ii) compounds with moderate permeability may require an intubation study if the dog colon and in silico models predict a marginally acceptable CR concentration-time profile; (iii) use a dose that approximates 1 h of the intended CR delivery rate; (iv) use the smallest volume possible; (v) define and record tubing placement; (vi) use a thermodynamically stable solution or/and suspension.

Gut Microbiome: Profound Implications for Diet and Disease.

The gut microbiome plays an important role in human health and influences the development of chronic diseases ranging from metabolic disease to gastrointestinal disorders and colorectal cancer. Of increasing prevalence in Western societies, these conditions carry a high burden of care. Dietary patterns and environmental factors have a profound effect on shaping gut microbiota in real time. Diverse populations of intestinal bacteria mediate their beneficial effects through the fermentation of dietary fiber to produce short-chain fatty acids, endogenous signals with important roles in lipid homeostasis and reducing inflammation. Recent progress shows that an individual's starting microbial profile is a key determinant in predicting their response to intervention with live probiotics. The gut microbiota is complex and challenging to characterize. Enterotypes have been proposed using metrics such as alpha species diversity, the ratio of Firmicutes to Bacteroidetes phyla, and the relative abundance of beneficial genera (e.g., Bifidobacterium, Akkermansia) versus facultative anaerobes (E. coli), pro-inflammatory Ruminococcus, or nonbacterial microbes. Microbiota composition and relative populations of bacterial species are linked to physiologic health along different axes. We review the role of diet quality, carbohydrate intake, fermentable FODMAPs, and prebiotic fiber in maintaining healthy gut flora. The implications are discussed for various conditions including obesity, diabetes, irritable bowel syndrome, inflammatory bowel disease, depression, and cardiovascular disease.

The impact of gastric pH, volume, and emptying on the food effect of ziprasidone oral absorption.

In a recent food effect clinical study, the authors concluded that a meal consisting of ≥500 kcal, regardless of fat content, produced the maximal bioavailability for ziprasidone. Using GastroPlus™, a commercially available pharmacokinetic simulation software, a semiphysiological model-a kind of physiologically based pharmacokinetic (PBPK) absorption model-was developed that could predict the concentration-time profiles when ziprasidone was administered with any one of the five test meals or fasting. Ziprasidone intravenous pharmacokinetics and oral absorption permeability were determined from clinical studies following the intravenous and duodenal infusion of ziprasidone to volunteers. From the detailed dietary information of each meal provided in the previously published food effect study, the stomach pH, volume, and gastric emptying could be predicted. Incorporating these meal-specific parameters into the model improved the predictions beyond the default fed/fasted parameters commonly used in the software. Compared to the default models, the improved models resulted in an improved prediction of the average ziprasidone concentration-time profile for each meal. Using this type of semiphysiological absorption model, we have shown that the dietary contents of the meals should be taken into account to predict food effects for ziprasidone and perhaps other BCS class I or II compounds.

Companion animal physiology and dosage form performance.

Among the most critical parameters for any drug candidate are tolerability, dose, solubility and permeability. For controlled release formulations, gastrointestinal transit is an added hurdle. While we might assume that intestinal transit is independent of the drug candidate, the relative importance of gastrointestinal transit time (GITT) depends directly on the other parameters. For example, a formulation of a drug with low solubility (LS) and/or low permeability (LP) characteristics might provide the required systemic concentrations when administered with food, but not if administered on an empty stomach. In the LS case, the drug may require the solubilizing effects of increased fluid and bile salts that accompany the meal. Likewise, a controlled release formulation of a drug with a region of preferred absorption may empty from the fasted stomach and move beyond the region before drug release is complete. Companion animals (e.g. cats and dogs) differ from humans and each other with respect to GITT, food effects, eating habit influences, breed and size variability, gastric pH, intestinal enzymes, GI permeability and absorption regions. This review examines how the anatomy and physiology of companion animals relates to the performance of orally administered immediate and controlled release formulations. Examples are presented of techniques used to predict the dose and acceptable solubility of drug candidates, and the performance of formulations in companion animals.

A computational model for particle size influence on drug absorption during controlled-release colonic delivery.

The effect of particle size on the percent drug absorbed is computationally modeled for controlled-release dosage forms that deliver drug particles to the colon. The relative benefit of reducing particle size is mapped on a diagram of the drug's absorption rate constant (estimated from rat intestinal perfusion, CACO-2 or human intubation permeation rates) versus the drug's solubility. Some drugs fall into a limit of high percentage absorption even with large particles such that particle size reduction has little impact. Another group of drugs is solubility limited such that even with small particles, absorption is negligible. Between the two regions, only drugs with sufficiently high absorption rates are influenced by the drug dissolution rate and thereby the particle size. The size of this region is a function of dosing rate. Comparisons between calculated particle size effects on colon absorption as a function of colon volume suggest caution when using animal models to predict bioavailability from colonic drug delivery. This volume dependence also suggests that the particle size influence will vary as a function of the digestive cycle.

A statistical method for the determination of absorption rate constant estimated using the rat single pass intestinal perfusion model and multiple linear regression.

The guide "Waiver of In Vivo Bioavailability and Bioequivalence Studies for Immediate Release Solid Dosage Forms Containing Certain Active Moieties/Active Ingredients Based on a Biopharmaceutical Classification System" (Rockville, MD: CDER, 2000) outlined non-in vivo tests of permeability that may satisfy the classification of a compound in the biopharmaceutical classification system. However, absent from that document were specific statistical methods to legitimatize the non-in vivo tests. This report describes the appropriate statistical treatment of absorption data, and recommends its adoption in the estimation of absorption and/or permeability measurements. The calculation of the absorption rate constants (k(a)) of ten compounds by a new multiple linear regression (MLR) method was completed after the separate perfusion of each compound through the rat single pass intestinal perfusion model (n = 3 rats per compound). Studentized residuals were evaluated to determine whether any statistically significant outliers were present in the data. The standard error of k(a) was estimated using variance components from the random effects model. The results were compared with the "traditional method" for k(a) calculations. Although both methods produced similar values of k(a), the MLR method's error estimate included multiple components of variability, which was largely ignored by the traditional method. The MLR method provided objective tests for outliers and achievement of steady-state. A preferred method for the statistical analysis of absorption data was demonstrated. These methods should be applied to all forms of permeability measurements, especially the non-in vivo measurements that classify a compound in the biopharmaceutical classification system.

Role of physiological intestinal water in oral absorption.

Water volume has impact when the compound has low aqueous solubility. For example, the absorption of compounds with a Biopharmaceutics Classification System class 2 or 4 is likely to be solubility-limited. Provided the formulation does not contribute to a dissolution-limited condition (e.g., particle size, Waterman and Sutton, J Control Release 86:293-304, 2003) and permeability is rapid, any impact on solubility factors in the gastrointestinal (GI) tract will directly impact the fraction absorbed These factors are in situ solubility, precipitation, and volume of water. Using GastroPlus, models were developed with literature values of water volume in the small (SIWV) and large (LIWV) intestines for several solubility limited compounds. One or more models were developed for the mean plasma concentration-time profile of each compound. The consistency of the models with known literature and experimental data for the compounds' solubility and precipitation was determined. The SIWV associated with best fits of solubility limited compounds averaged about 130 ml, with a range of 10-150 ml in the fasted state. The average LIWV in the fasted state was about 10 ml and ranged as large as 125 ml. The wide range of individual LIWV values is likely due to variability in pharmacokinetics, permeability, GI transit, and the observation that data set was collected during a "snapshot in time". The preferred values of 10% organ volume for small intestine and 1-10% organ volume for large intestine are recommended in lieu of the GastroPlus default values of 40% and 10%, respectively.

Fed and fasted gastric pH and gastric residence time in conscious beagle dogs.

The gastric pH values are controversial in the literature. Some suggest the dog gastric pH is higher than human and dog gastric pH after fed with particular diet is uncertain. Gastric pH in 16 male beagle dogs was measured using Bravo pH telemetry system. For the fed study, the dogs received 10 or 200 g of dog dry food (5L18) 15 min before dosing the Bravo pH capsule, followed by a 50 mL of water to aid in swallowing. It was surprising to find a small, but statistically significantly lower pH in the fed compared to the fasted stomach. The average gastric pH in fasted dogs was 2.05 and 1.08 and 1.26 for 10 and 200 g fed dogs. The average gastric emptying time of the capsule was 1.4, 9.4 and 20 h for fasted, 10 g fed and 200 g fed dogs, respectively. The inter-individual variability was higher in fasted dogs than in fed dogs. The results showed the gastric pH in each colony of dogs can be different from reported values in the literature. It emphasizes that the importance of measuring the pH in each colony when dogs are used to evaluate pharmacokinetics of pH sensitive drugs or formulations.

An automated process for building reliable and optimal in vitro/in vivo correlation models based on Monte Carlo simulations.

Many mathematical models have been proposed for establishing an in vitro/in vivo correlation (IVIVC). The traditional IVIVC model building process consists of 5 steps: deconvolution, model fitting, convolution, prediction error evaluation, and cross-validation. This is a time-consuming process and typically a few models at most are tested for any given data set. The objectives of this work were to (1) propose a statistical tool to screen models for further development of an IVIVC, (2) evaluate the performance of each model under different circumstances, and (3) investigate the effectiveness of common statistical model selection criteria for choosing IVIVC models. A computer program was developed to explore which model(s) would be most likely to work well with a random variation from the original formulation. The process used Monte Carlo simulation techniques to build IVIVC models. Data-based model selection criteria (Akaike Information Criteria [AIC], R2) and the probability of passing the Food and Drug Administration "prediction error" requirement was calculated. To illustrate this approach, several real data sets representing a broad range of release profiles are used to illustrate the process and to demonstrate the advantages of this automated process over the traditional approach. The Hixson-Crowell and Weibull models were often preferred over the linear. When evaluating whether a Level A IVIVC model was possible, the model selection criteria AIC generally selected the best model. We believe that the approach we proposed may be a rapid tool to determine which IVIVC model (if any) is the most applicable.

Dog colonoscopy model for predicting human colon absorption.

PURPOSE: This study was conducted to develop and validate a dog colon model that predicts colon permeability in humans. METHODS: The following compounds were studied: Class 1 highly soluble (HS)/highly permeable (HP): aminophylline, propranolol, CP-409092; Class 2 LS/HP: nifedipine; trovafloxacin, sertraline; Class 3 HS/LP: azithromycin, atenolol, CP-331684, CP-424391; Class 4 LS/LP: CJ-13610. Administration to dogs was made 30 cm cranial to the anal sphincter with a lubricated Schott Model VFS-5 flexible endoscope. The bioavailability of the compound following the colon administration in dogs, relative to the same formulation administered orally (relative bioavailability), was determined. RESULTS: Except for atenolol, a small hydrophillic molecule, the relative bioavailability from administration to the colon of the dog correlated well with the following compound properties: high solubility and high, passive permeability > high solubility, low permeability > low solubility, high, passive permeability approximately low solubility, low permeability. CONCLUSION: The dog colon model is proposed as a surrogate for human intubation studies when the controlled release candidate falls in BCS Classes 2 (LS/HP), 3 (HS/LP), and 4 (LS/LP). However, no human intubation or dog colon studies are required for Class 1 (HS/HP), as these compounds are likely to be well absorbed from the colon.