Simultaneous Determination of Procaspase Activating Compound 1 and Permeability Markers in Intestinal Perfusion Samples and Application to a Rat Intestinal Absorption Study

A sensitive and simple HPLC method for simultaneous determination of PAC-1 (first procaspase-activating compound), phenol red, and permeability markers (carbamazepine and furosemide) in perfusion samples was developed and validated to assess intestinal absorption of PAC-1 using single-pass intestinal perfusion technique (SPIP) in rats. The chromatographic separation was carried out on a Kromasil C₁₈ column (150 mm × 4.6 mm, 5 μm) with acetonitrile–methanol–30 mmol L⁻¹ phosphate buffer (pH 3.0, 25:10:65, v/v/v) as mobile phase at a flow rate of 1.0 mL min⁻¹, and the wavelength of the UV detector was set at 281 nm. The calibration curves were linear in the ranges of 2.40–48.0 μg mL⁻¹ for PAC-1; 3.60–72.0 μg mL⁻¹ for carbamazepine; 3.20–64.0 μg mL⁻¹ for furosemide, and 4.80–96.0 μg mL⁻¹ for phenol red (r > 0.999). Both the intra- and inter-day precisions (RSD%) of all analytes were less than 6.8 % at three concentration levels, while accuracy ranged from 95.4 to 104.5 %. Data obtained in all method validation studies indicated that the method was suitable for the intended purpose. The effective permeability values (P _eff) considering water flux with the help of non-permeable marker phenol red was calculated to be 0.42 × 10⁻⁴, 0.62 × 10⁻⁴, 0.32 × 10⁻⁴ cm s⁻¹ for PAC-1; 0.72 × 10⁻⁴, 0.77 × 10⁻⁴, 0.52 × 10⁻⁴ cm s⁻¹ for carbamazepine; 0.20 × 10⁻⁴, 0.16 × 10⁻⁴, 0.12 × 10⁻⁴ cm s⁻¹ for furosemide in duodenum, jejunum and ileum, respectively. The P _eff value can be increased by co-perfusion with verapamil, indicating that absorption of PAC-1 is efficiently transported by P-glycoprotein (P-gp) in the gut wall.

     

Recent Advancements in Using Polymers for Intestinal Mucoadhesion and Mucopenetration

Oral administration of actives is the most desired form of delivery, but the formulations need to overcome a variety of barriers including the intestinal mucus. This feature article summarizes the developments from the past 2–3 years in this context focusing on polymer‐based formulations. The progress in assembling mucopenetrating nanoparticles is outlined considering coatings using noninteracting polymers as well as virus‐like particles and charge‐shifting particles. Next, polymers and their modification to enhance mucoadhesion are discussed, followed by providing examples of double‐encapsulation systems that aim to combine mucopenetration with mucoadhesion in the same formulation. Finally, a short outlook is provided highlighting a few of the most pressing challenges to address.     

Intestinal metabolism of Polygonum cuspidatum in vitro and in vivo

Rhizoma et Radix Polygoni Cuspidati (RRPC) is commonly prescribed for the treatment of amenorrhea, arthralgia, jaundice and abscess in traditional Chinese medicine. Previous pharmacological studies have indicated that polyphenols are the main pharmacological active ingredients in RRPC. Meanwhile, the poor bioavailability of polyphenols in RRPC implies that those components are probably metabolized by intestinal bacteria before absorption. However, there is rather limited information about RRPC''s metabolites produced by intestinal bacteria and the intestinal absorbed constituents. In the present study, the metabolites were characterized after the aqueous extract of RRPC was incubated with the crude enzyme of human intestinal bacteria in vitro. The metabolic characteristics of glycosides in RRPC were figured out by comparing the metabolic profiles of emodin‐8‐O‐β‐d ‐glucopyranoside and polydatin between aqueous extract of RRPC and equivalent amounts of these two glycosides. The transitional constituents absorbed into blood were investigated in rats via intraduodental administration and portal vein intubation. A total of 38 prototype components and 43 metabolites were detected and characterized in vivo. The overall results demonstrated that the intestinal bacteria played an important role in the metabolism of RRPC, and the main metabolic pathways were hydrolysis in vitro, glucuronidation and sulfation in vivo.     

Intestinal absorption of dolichol from emulsions and liposomes in rats

The intestinal absorption of dolichol from various dosage forms was investigated using the intestinal loop and everted sac methods in the rat. The in situ loop experiments showed that the absorption of dolichol from a triglyceride emulsion was dependent on the chain-length of the triglyceride; the absorption from a tri-n-butyrin emulsion in 1 h was 18.0% of the dose; and the absorption from an HCO-60 suspension was 4.3%. The liposomal preparation enhanced the absorption up to 39.1% of the dose. In in vitro experiments, 25.0% and 13.2% of dolichol were taken up by everted sacs of the jejunum and the ileum, respectively. On the other hand, phospholipids composing liposomes were not absorbed under these conditions. The above results suggest that the absorption mechanism from liposomal preparations may be as follows: dolichol is released from the liposomes into the aqueous phase adjacent to the surface of the intestine and is subsequently partitioned into the intestinal tissue.     

Strawberry Decreases Intraluminal and Intestinal Wall Hydrolysis of Testosterone Undecanoate

Male hypogonadism is often treated by testosterone (T) replacement therapy such as oral administration of the ester prodrug, testosterone undecanoate (TU). However, the systemic exposure to T following oral TU is very low due to esterase-mediated metabolism, particularly in the small intestine. The aim of this work was to examine the esterase-inhibitory effect of natural fruit extract of strawberry (STW) on the intestinal degradation of TU as a potential approach to increasing the oral bioavailability of T. Herein, the hydrolysis of TU was assessed in fasted state simulated intestinal fluid with added esterase activity (FaSSIF/ES) and Caco-2 cell homogenates in the presence of STW extract. It is noteworthy that STW substantially inhibited the degradation of TU in FaSSIF/ES and Caco-2 cell homogenates at concentrations that could be achieved following oral consumption of less than one serving of STW fruit. This can significantly increase the fraction of unhydrolyzed TU in the intestinal lumen as well as in enterocytes. In addition, it was demonstrated that TU has high intestinal lymphatic transport potential as the association of TU with plasma-derived human chylomicrons was in the range of 84%. Therefore, oral co-administration of TU with STW could potentially increase the intestinal stability of TU and consequently the contribution of lymphatically delivered TU to the systemic exposure of T in vivo.     

大肠癌与七种元素关系的分析

用原子吸收光谱法测定了38例大肠癌组织及其边缘组织中铬、锰、锌、铁、镁、钙、铜的含量.结果表明,癌组织中锰含量(0.281±0.179)×10-6,镁含量(76.85±17.66)×10-6,高于其边缘组织(P＜0.01).七种元素含量与大肠癌的病理类型及分化程度无关.提示,大肠癌的发生与锰、镁的含量变化有关.     

LC Evaluation of Intestinal Transport of Praziquantel

Praziquantel (PZQ) is a highly lipophilic drug with low aqueous solubility. Despite this, it is well absorbed from the gastrointestinal tract. In this study, a simple LC method was developed and validated, in order to monitor the concentration of PZQ in TC-199 buffer in vitro, in the rat everted gut sac absorption model. PZQ was analyzed by a reversed-phase LC method with an isocratic mobile phase containing acetonitrile and water in the proportions 45:55. The flow-rate was 1 mL min⁻¹ and PZQ was determined by measuring absorbance at 215 nm, at 25 °C. The method was found to be specific, as none of the components of TC-199 or intestinal sac artefacts interfered with the drug peak. Recovery was within acceptable statistical limits. The limit of detection was 0.54 μg mL⁻¹ and the limit of quantitation was 1.63 μg mL⁻¹. The calibration curve was found to be linear in the concentration range of 10–90 μg mL⁻¹ PZQ. The proposed method was found to be rapid and selective and hence can be applied in the monitoring of the absorption of PZQ in in vitro everted gut sac absorption studies.     

LC Evaluation of Intestinal Transport of Praziquantel

Praziquantel (PZQ) is a highly lipophilic drug with low aqueous solubility. Despite this, it is well absorbed from the gastrointestinal tract. In this study, a simple LC method was developed and validated, in order to monitor the concentration of PZQ in TC-199 buffer in vitro, in the rat everted gut sac absorption model. PZQ was analyzed by a reversed-phase LC method with an isocratic mobile phase containing acetonitrile and water in the proportions 45:55. The flow-rate was 1 mL min⁻¹ and PZQ was determined by measuring absorbance at 215 nm, at 25 °C. The method was found to be specific, as none of the components of TC-199 or intestinal sac artefacts interfered with the drug peak. Recovery was within acceptable statistical limits. The limit of detection was 0.54 μg mL⁻¹ and the limit of quantitation was 1.63 μg mL⁻¹. The calibration curve was found to be linear in the concentration range of 10–90 μg mL⁻¹ PZQ. The proposed method was found to be rapid and selective and hence can be applied in the monitoring of the absorption of PZQ in in vitro everted gut sac absorption studies.

     

Deoxycholic Acid Modulates Cell-Junction Gene Expression and Increases Intestinal Barrier Dysfunction

Diet-related obesity is associated with increased intestinal hyperpermeability. High dietary fat intake causes an increase in colonic bile acids (BAs), particularly deoxycholic acid (DCA). We hypothesize that DCA modulates the gene expression of multiple cell junction pathways and increases intestinal permeability. With a human Caco-2 cell intestinal model, we used cell proliferation, PCR array, biochemical, and immunofluorescent assays to examine the impact of DCA on the integrity of the intestinal barrier and gene expression. The Caco-2 cells were grown in monolayers and challenged with DCA at physiological, sub-mM, concentrations. DCA increased transcellular and paracellular permeability (>20%). Similarly, DCA increased intracellular reactive oxidative species production (>100%) and accompanied a decrease (>40%) in extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathways. Moreover, the mRNA levels of 23 genes related to the epithelial barrier (tight junction, focal adhesion, gap junction, and adherens junction pathways) were decreased (>40%) in (0.25 mM) DCA-treated Caco-2 cells compared to untreated cells. Finally, we demonstrated that DCA decreased (>58%) the protein content of occludin present at the cellular tight junctions and the nucleus of epithelial cells. Collectively, DCA decreases the gene expression of multiple pathways related to cell junctions and increases permeability in a human intestinal barrier model.     

配糖蛋白B肠溶微胶囊的研究

采用双乳液法研制配糖蛋白Ｂ的肠溶微胶囊，考察了微胶囊的形态、粒径及其分布，在摸拟肠液的缓冲液中进行溶解释放试验。配制了两种口服微胶囊混悬制剂，并考察了它的稳定性。     

In Vitro Gastric and Intestinal Digestions of Pulsed Light-Treated Shrimp Extracts

Pulsed ultraviolet light (PUV), a novel technology most commonly used for microbial inactivation, has recently been employed to effectively mitigate food allergens in peanuts, soybean, shrimp, and almond. Putative mechanisms for the efficacy of PUV in reducing allergen reactivity include photothermal, photochemical, and photophysical effects. To date, there are no published data highlighting the effects of in vitro simulated gastric and intestinal digestion on the stability of PUV reduced allergen reactivity of food. In this study, PUV-treated shrimp extracts were subjected to simulated gastric fluid containing pepsin and simulated intestinal fluid containing trypsin and chymotrypsin, and then tested for changes in allergen potency. SDS-PAGE showed no major band deviation between undigested and digested PUV-treated shrimp extracts. IgE binding to tropomyosin remained markedly decreased as seen in Western blot analysis. Total shrimp allergen reactivity remained unchanged following in vitro peptic digestion and was markedly reduced following in vitro intestinal digestion as illustrated in indirect ELISA. The PUV reduced shrimp allergens remained at a low level under the in vitro simulated digestive conditions. The results inferred that PUV could be a potential method to create less allergenic shrimp products that would remain at a low allergen level under human gastric and intestinal digestive conditions.     

QSAR Study and VolSurf Characterization of Human Intestinal Absorption of Druge

The prediction of human intestinal absorption is a major goal in the design,optimization,and selection of candidates for the develoment of oral drugs.In this study,a computerized method(VolSurf with GRID) was used as a novel tool for predicting human intestinal absorption of test compound,and for determining the critical molecular properties needed for human intestinal absorption.The tested molecules consisted of 20 diverse drug-like compounds.Partial least squares(PLS) discriminant analysis was used to correlate the experimental data with the theoretical molecular properties of human intestinal absorption.A good correlation(r^2=0.95,q^2=0.86) between the molecular modeling results and the experimental data demonstrated that human intestinal absorption could be predicted from the three-dimensional(3D) molecular structure of a compound .Favorable structureal properties identified for the potent intestinal absorption of drugs included strong imbalance between the center of mass of a molecule and the barycentre of its hydrophilic and hydrophobic regions and a definitive hydrophobic region as well as less hydrogen bonding donors and acceptors in the molecule.     

Intestinal absorption of tellurium studied with stable tracers

An investigation on tellurium metabolism by administration of stable tellurium isotopes¹²⁴Te and¹²⁶Te has been performed. Fractional intestinal absorption was determined in rabbits by the double tracer technique. The investigated subjects were given an enriched solution of one tellurium isotope orally and a few minutes later an enriched solution of the other isotope intravenously. The¹²⁴Te and¹²⁶Te contents in plasma samples were determined by proton nuclear activation. The methodology described offers a means to study tellurium metabolism in humans without radiation risk.     

Determination of Ethynylestradiol and Norethindrone in Synthetic Intestinal Fluid and in Timed-Release Oral Formulations

Abstract

High performance liquid chromatographic methods are described for the rapid assay of the antifertility steroids ethynylestradiol and norethindrone in sustained-release oral formulations. A novel method is reported for isolation of these hormones from the synthetic intestinal fluid used for in-vitro release rate studies.     

海藻酸钠-壳聚糖微胶囊作为肠道内生化微反应器的研究

基因工程技术的发展使蛋白、多肽类高值生化药物的大规模生产得以实现并用于临床^[1].但目前存在产物分离、纯化工艺复杂、成本高等问题.因此,研制一种无需分离纯化、低成本的肠道内生化微反应器作为基因工程药物释放系统具有实际应用意义(例如将基因工程微生物包埋在具有半透性高分子膜的微胶囊中,口服后微囊化活细胞在肠道内生长并分泌有治疗作用的基因工程药物而达到治疗目的^[2]).本文以酵母菌Pichia pastoris GS115为模型菌株,以海藻酸钠-壳聚糖(A lginate-chitosan,AC)微胶囊为载体,考察了AC微囊化酵母菌在模拟胃肠液中的形态、膨胀性能、酵母菌存活率及小鼠口服后肠道黏膜粘附性能,初步证明AC微囊化基因工程酵母菌作为肠道生化微反应器是可行的.