Impact of active pharmaceutical ingredient variables and oleaginous base on the in vitro drug release from ophthalmic ointments: An investigation using dexamethasone as a model drug.

The present study systematically investigates the impact of active pharmaceutical ingredient (API) variables and oleaginous base characteristics on the in vitro release (IVR) performance of ophthalmic ointments, utilizing dexamethasone as a model drug. The interplay between selected attributes (i.e., particle size distribution, crystallinity, and polymorphic form for API, and rheological factors for compendial-grade white petrolatum) and IVR performance was investigated. APIs from different vendors exhibited variations in crystallinity and polymorphism. Ointments containing amorphous dexamethasone presented higher release amounts/rates compared to crystalline counterparts, emphasizing the role of physical state in release kinetics. Variations in particle size of this lipophilic API (5.4 - 21.2 µm) did not appear to impact IVR performance significantly. In contrast, white petrolatum's rheological attributes, which varied substantially within USP-grade petrolatum, were found to critically affect the drug release rate and extent of the ointment. The study's comprehensive analysis establishes a coherent connection between the quality attributes of both API and petrolatum and IVR, delineating their intricate interdependent effects on ophthalmic ointment performance. These findings provide reference to formulation design, quality control, and regulatory considerations within the pharmaceutical industry, fostering a robust foundational understanding of commonly overlooked quality attributes in ophthalmic ointments.

Impact of Apparatus and Adapter on In vitro Drug Release of Ophthalmic Semisolid Drug Products.

PURPOSE: In vitro release testing (IVRT) is a widely used tool for evaluating the quality and performance of drug products. However, standardized sample adaptors or drug release apparatus setups for IVRT studies are still lacking for ophthalmic ointments. The aim of this study was to provide a better understanding of the impact of apparatus and sample adaptor setups on IVRT of ophthalmic ointments. METHODS: Dexamethasone (DEX), a steroidal ingredient commonly used in ophthalmic drug products, was selected as a model drug. Ointments were prepared by mixing DEX in white petrolatum using a high shear mixer. A novel two-sided adapter was developed to increase the drug release surface area. DEX ointment was placed in one-sided or two-sided release adaptors coupled with 1.2 µm polyethersulfone membrane, and the drug release was studied in different USP apparatuses (I, II, and IV). RESULTS: The sample adaptor setups had a minimal impact on cumulative drug release amount per area or release rate while USP IV apparatus with agitated flow enhanced drug release rates. The USP apparatus I with a two-sided semisolid adapter, which uses membranes on both sides, showed dramatically higher cumulative drug release and discriminative release profiles when evaluating ophthalmic formulations. CONCLUSIONS: USP apparatuses and sample adaptors are critical considerations for IVRT. Two-sided semisolid adapter provides higher cumulative release, facilitating the discrimination between low drug content ophthalmic ointment formulations with good sensitivity and repeatability without affecting the drug release rate.

Impact of Membranes on In Vitro Release Assessment: a Case Study Using Dexamethasone.

In vitro release studies are commonly used to assess the product performance of topical dosage forms. In such studies, the mass transport of drugs through synthetic membranes into a receiving chamber filled with a release medium is measured. The release medium is also passed through filtration membranes prior to chromatographic analysis. There are no official guidelines directing membrane selection for in vitro release studies or for filtration. Considering the diversity in membrane materials and their physical properties, the aim of this study was to investigate membrane-drug binding and the effect of various membranes on the release performance of a model drug dexamethasone (DEX) using USP dissolution apparatus IV. Seven membranes of different pore sizes (0.45 and 1.2 µm) and materials (cellulose acetate, polyethersulfone, and nylon) were assessed. Two different methods, syringe filter and 24-h incubation, were used for the determination of membrane-drug binding effects at low drug concentrations and saturated concentration conditions. Cellulose acetate and nylon membranes showed significant drug binding after 24-h incubations at both drug concentrations. DEX diffusion through membranes was significantly slowed down in all the tested membranes when compared with DEX solution without membranes. The extent of the retardation varied due to the differences in membrane structures. In conclusion, materials and sources of membranes affected drug dissolution profiles and the results showed membrane-drug binding effects. Proper selection of membranes with low drug binding ability and low diffusion resistance is essential to ensure appropriate and reproducible in vitro release assessments and filtration studies. Graphical Abstract.

Influence of in vitro release methods on assessment of tobramycin ophthalmic ointments.

The current investigation was carried out to identify appropriate parameters for measuring the in vitro release of tobramycin (TOB) ophthalmic ointments and to evaluate the feasibility of in vitro release testing methods to assess the product performance. Drug release was assessed using USP dissolution apparatus IV and a modified USP dissolution apparatus I with simulated tear solution (STS) as the dissolution medium. The study variables included temperature, membrane material, source and pore size. The results demonstrated a significant influence of the membrane source and pore size on the release of TOB from the ointments. A dissolution medium temperature of 40 °C was found to be appropriate for the release studies. Both of the apparatuses were able to discriminate between the release profiles of ointments with different physicochemical/rheological properties. Maximum release rate of TOB was observed in the first hour which followed a logarithmic time dependent release. The correlation between the release rates/amounts and yield stress of the ointments was observed in both the dissolution apparatuses. These results support a rational approach to guide the in vitro release testing of TOB ophthalmic ointments.

Kaempferia parviflora Nanosuspension Formulation for Scalability and Improvement of Dissolution Profiles and Intestinal Absorption.

Kaempferia parviflora (KP) is an herbal medicine for enhancement of physical fitness and male sexual function improvement with low oral absorption of the main active compounds, methoxyflavones. The purpose of this study is to optimize the preparation of nanosuspensions of KP extract for enhancing intestinal absorption using antisolvent precipitation technique which is an accessible nanomanufacturing methodology in the small industrial factory. Nanosuspensions were prepared using various types and concentrations of stabilizers. Then, the dry powder of KP nanosuspension was produced by spray drying. Its dissolution rate was determined using USP dissolution apparatus II. The rat everted intestinal sac was tested to confirm the improvement of intestinal absorption of KP nanosuspension. The result showed that 3% sodium lauryl sulfate (SLS) was the optimal condition for covering the nano-size of KP nanosuspension. KP nanosuspensions had particle sizes ranging from 100 to 300 nm with narrow size distribution (PDI < 0.60) and zeta potential at - 58 to - 70 mV. These characteristics were stable at 4°C and 25°C/60%RH for 1-month storage. Its methoxyflavones content also unchanged at 4°C and 25°C/60%RH for 1-month storage. KP nanosuspension released > 80% of the methoxyflavones within 30 min both in 0.1 N HCl and 0.01 M phosphate buffer solution (pH 6.8). Moreover, the developed nanosuspension dramatically improved the rat intestinal absorption about 10-fold. Therefore, the KP nanosuspension was successfully prepared. It has relatively high stability, fast dissolution rate, and high intestinal absorption.

In vivo effect of Kaempferia parviflora extract on pharmacokinetics of acetaminophen.

Kaempferia parviflora is widely used as a food supplement and a herbal medicine for vitalization. Previous study has shown that K. parviflora had CYP2E1 inducer activity. It is likely to affect the metabolism of CYP2E1 substrates such as acetaminophen which is a common household pain relief medicine. This study investigated the possible pharmacokinetic interaction between K. parviflora and acetaminophen in rats. Acetaminophen (100 mg/kg, p.o) was administered to rats for nine consecutive days. On days 4-9, K. parviflora extract (250 mg/kg, p.o) was given to the acetaminophen-treated rats. After co-administration with K. parviflora, the concentrations of acetaminophen during day 5-8 markedly decreased compared with acetaminophen-only group. At day 9, the pharmacokinetic parameters of acetaminophen in the presence of K. parviflora extract also decreased, including area under the concentration-time curve (from 1.68 ± 0.16 to 0.34 ± 0.04 mg.min/mL), the maximum concentration (from 19.10 ± 1.90 to 4.48 ± 0.56 µg/mL), and half-life (from 21.29 ± 1.36 to 10.81 ± 1.24 min). In addition, clearance and the elimination rate constant of acetaminophen were significantly increased (from 0.003 ± 0.000 to 0.006 ± 0.001 L/min and 0.03 ± 0.00 to 0.07 ± 0.01 min-1, respectively) in the presence of K. parviflora extract. These findings provide the data for in vivo herb-drug interaction between K. parviflora extract and acetaminophen. Therefore, the concomitant use of K. parviflora as a food supplement and acetaminophen should occasion therapeutic and safety concerns.

Glucose Tolerance Test and Pharmacokinetic Study of Kaempferia parviflora Extract in Healthy Subjects.

Kaempferia parviflora Wall. ex Baker (KP), Krachaidam in Thai or Thai ginseng, is a herbal medicine that has many potential pharmacological effects. The effect of KP extract on blood glucose level in rodent was reported. This study focused on the oral glucose tolerance test and pharmacokinetic study in healthy volunteers administered with KP extract (90 and 180 mg/day, placebo). The oral glucose tolerance tests were performed at baselines and 28-days of administration. The pharmacokinetics were determined after a single dose administration of the tested products using 3,5,7,3',4'-pentamethoxyflavone (PMF) and 5,7,4'-trimethoxylflavone (TMF) as markers. The results showed that glucose metabolism via oral glucose tolerance test was not affected by KP extract. Blood glucose levels of volunteers at 120 min after glucose loading were able to be returned to initial levels in placebo, KP 90 mg/day, and KP 180 mg/day groups both at baseline and 28-days of administration. The results of the pharmacokinetic study revealed that only TMF and PMF, but not 5,7-dimethoxyflavone (DMF) levels could be detected in human blood. The given doses of KP extract at 90 and 180 mg/day showed a linear dose-relationship of blood PMF concentration whereas blood TMF was detected only at high given dose (180 mg/day). The half-lives of PMF and TMF were 2-3 h. The maximum concentration (Cmax), area under the curve of blood concentration and time (AUC), and time to maximum concentration (Tmax) values of PMF and TMF estimated for the 180 mg/day dose were 71.2 ± 11.3, 63.0 ± 18.0 ng/mL; 291.9 ± 48.2, 412.2 ± 203.7 ng∙h/mL; and 4.02 ± 0.37, 6.03 ± 0.96 h, respectively. PMF was quickly eliminated with higher Ke and Cl than TMF at the dose of 180 mg/day of KP extract. In conclusion, the results demonstrated that KP extract had no effect on the glucose tolerance test. In addition, this is the first demonstration of the pharmacokinetic parameters of methoxyflavones of KP extract in healthy volunteers. The data suggest the safety of the KP extract and will be of benefit for further clinical trials using KP extract as food and sport supplements as well as a drug in health product development.

Pharmacokinetic interaction between Kaempferia parviflora extract and sildenafil in rats.

Kaempferia parviflora (KP) is a plant widely used in Southeast Asia. Its major compounds are 3,5,7,3',4'-pentamethoxyflavone (PMF), 5,7,4'-trimethoxylflavone (TMF), and 5,7-dimethoxyflavone (DMF). This study investigated the effect of KP extract on the blood levels and pharmacokinetics of sildenafil co-administration in rats. Rats were randomly assigned to four groups. Groups 1, 2, and 3 were given sildenafil 20 mg/kg daily for 9 days. On days 4-9 of each treatment period, the treated rats received KP extract (250 mg/kg) and vehicle (groups 2 and 3, respectively). Group 4 received KP extract only (250 mg/kg daily for 9 days). Daily blood concentrations of sildenafil, PMF, TMF, and DMF were determined by HPLC to evaluate the daily blood level interactions. Additional blood samples were collected at various times on the last day of treatment to evaluate the pharmacokinetic interactions. The KP extract decreased blood levels of sildenafil on the first day of co-administration by 95 % but the percentage reduction was insignificant on subsequent days. When co-administered with KP extract, the area under the curve (AUC), maximum concentration (C max), and half-life (T 1/2) of sildenafil were decreased by 60-65, 40-52, and 32-54 %, respectively, with the elimination rate constant (K e) increased by 37-77 %. In addition, PMF, TMF, and DMF concentrations and their AUC, C max, T max, K e, and T 1/2 values were changed after co-administration of KP extract and sildenafil.

Novel formulation strategies for enhancing oral delivery of methoxyflavones in Kaempferia parviflora by SMEDDS or complexation with 2-hydroxypropyl-ß-cyclodextrin.

The Kaempferia parviflora (KP) plant contains several methoxyflavones including 5,7-dimethoxyflavone (DMF), 5,7,4'-trimethoxyflavone (TMF), and 3,5,7,3',4'-pentamethoxyflavone (PMF). Ethanolic extracts of KP have shown various pharmacological effects and have been used as an aphrodisiac, a antimicrobial agent and for the treatment of inflammation, and peptic ulcers. Given its poor water solubility and low oral bioavailability (1-4%), there are limitations to the utilization of KP. Accordingly, self-microemulsifying drug delivery system (SMEDDS) and cyclodextrin (CD) complex formulations were developed to improve the oral absorption of methoxyflavones. Polyoxyethylene castor oil (53.3%), propylene glycol (26.7%), and triglyceride of coconut oil (20%) were combined to form KP-SMEDDS. A complex of 2-hydroxypropyl-ß-cyclodextrin (2-HP-ß-CD) and KP was prepared by lyophilization. The developed formulations were then evaluated for their physicochemical properties, in vitro dissolution tests, permeability through Caco-2 cells, and in vivo oral absorption in rats by using PMF, TMF, and DMF as the markers for quantitation. The results showed that KP-SMEDDS and KP-2-HP-ß-CD complex improved the dissolution rate of methoxyflavones in both 0.1N HCl and 0.2M PBS pH 6.8 compared to KP dissolved in a solution of propylene glycol, PEG 400, ethanol, and water. KP-SMEDDS and KP-2-HP-ß-CD formulations showed about 10- and 3.5-fold greater Papp values of methoxyflavones in Caco-2 cells. The oral bioavailability values of KP-SMEDDS formulations were higher than those of KP (25.38-, 42.00-, and 26.01-fold for PMF, TMF, and DMF, respectively). For the KP-2-HP-ß-CD complex, oral bioavailability values were 21.63-, 34.20-, and 22.90-fold greater than those of KP, respectively. Therefore, these two novel formulations, KP-SMEDDS and KP-2-HP-ß-CD, were successfully developed to improve the dissolution rate, drug permeability through Caco-2 cells and oral bioavailability of methoxyflavones in KP.

Pharmacokinetics, bioavailability, tissue distribution, excretion, and metabolite identification of methoxyflavones in Kaempferia parviflora extract in rats.

Kaempferia parviflora (KP) is an herbal plant in the family of Zingiberaceae. KP mainly contains methoxyflavones, especially 5,7-dimethoxyflavone (DMF), 5,7,4'-trimethoxyflavone (TMF), and 3,5,7,3',4'-pentamethoxyflavone (PMF). The present study was designed to characterize the pharmacokinetics, including bioavailability, distribution, excretion, and identification of metabolites after administration of a KP ethanolic extract. Male rats were orally or intravenously administered a 250 mg/kg concentration of a KP extract, and blood samples were obtained at selected times to determine pharmacokinetic parameters of PMF, TMF, and DMF. For distribution and excretion studies, the organs, urine, and feces samples were collected at various times after oral administration of a larger (750 mg/kg) dose of KP extract. Methoxyflavones in the biological samples were quantified by high-performance liquid chromatography-UV, and the metabolites in urine and feces were further identified by using liquid chromatography-tandem mass spectrometry. After oral administration, concentrations of the three methoxyflavones quickly approached their maximal concentration, ranging from 0.55 to 0.88 µg/ml within 1 to 2 h after administration, and then were gradually excreted with half-lives of 3 to 6 h. The methoxyflavones showed low oral bioavailability of 1 to 4%. Three methoxyflavones were detected at their highest levels in liver followed by kidney. They were also found in lung, testes, and brain. After absorption, organ distribution, and metabolism, the components of KP were mainly eliminated through urine in the forms of demethylated, sulfated, and glucuronidated products and as demethylated metabolites in the feces. The parent compounds were found to have 0.79, 1.76, and 3.10% dose recovery in urine and 1.06, 1.77, and 0.96% dose recovery in feces for PMF, TMF, and DMF, respectively. These studies are the first to describe the pharmacokinetics of KP extract to provide the information on blood and tissue levels.

Modulatory effects of Kaempferia parviflora extract on mouse hepatic cytochrome P450 enzymes.

ETHNOPHARMACOLOGICAL RELEVANCE: Kaempferia parviflora is a herbal plant, the extracts of which are commonly used as alternative medicines. It widely uses as aphrodisiac, anti-inflammation, anti-microbacterial, and anti-peptic ulcer. AIM OF THE STUDY: In order to obtain an effective utilization and safety of the herb, the influence of Kaempferia parviflora on hepatic CYP450 metabolizing enzymes including CYP1A1, CYP1A2, CYP2B, CYP2E1, and CYP3A was investigated. MATERIALS AND METHODS: The impact of Kaempferia parviflora on CYP450 both in vitro and in vivo was examined by using ethoxyresorufin O-dealkylation, methoxyresorufin O-dealkylation, pentoxyresorufin O-dealkylation, p-nitrophenol hydroxylation, and erythromycin N-demethylation assays, respectively. RESULTS: In vitro studies using non-induced mouse hepatic microsomes in the presence or absence of Kaempferia parviflora extract showed that Kaempferia parviflora extract altered CYP1A1, CYP1A2, CYP2B, and CYP2E1 activities by non-competitive, mixed-competitive, competitive, and uncompetitive mechanisms, respectively. Among these enzymes, CYP1A2 was affected by Kaempferia parviflora based on the highest value of V(max) (15.276±0.206 nmol/min) and lowest of K(i) value (0.008±0.002 µg/ml). In addition, the plant extract also modulated CYP2B activity based on the low K(m) value (1.599±0.147 pmol). For in vivo studies, mice were orally treated with 250 mg/kg of Kaempferia parviflora extract for 7, 14, and 21 days. The results demonstrated that Kaempferia parviflora extract significantly induced CYP1A1, CYP1A2 enzyme activities following short-term treatment. CYP2B enzyme activities were markedly increased all Kaempferia parviflora extract treatment timepoints, whereas Kaempferia parviflora extract significantly enhanced CYP2E1 activity only after long-term treatment. However, Kaempferia parviflora extract did not affect the CYP3A enzyme activity. CONCLUSIONS: Kaempferia parviflora extract modulated several CYP450 enzyme activities, thus, its utilization with drugs or other herbs should raise concern for potential drug-herb interactions.