Empiric “Three-in-One” Bismuth Quadruple Therapy for Second-Line Helicobacter pylori Eradication: An Intervention Study in Southern Italy

The eradication of Helicobacter pylori (H. pylori) may be difficult due to antibiotic resistance. Indeed, after one failure, a second-line therapy is needed and a bismuth containing quadruple therapy (BQT) with a three-in-one capsule formulation is becoming very popular. Therefore, we aimed to evaluate effectiveness and safety of BQT as a second-line therapy. We recruited consecutive patients with one therapy failure. For ten days patients received the three-in-one BQT Pylera® therapy, in combination with a proton-pump inhibitor (PPI), decided at the choice of the investigator, at full dose bid. The eradication rate was calculated by intention-to-treat (ITT) and per-protocol (PP)analyses and 95% confidence intervals (CI) were calculated. Seventy-three patients were recruited, 41 females and 32 males (mean age 53.0±13.1 years). Fifty-five patients failed triple therapy with amoxicillin and clarithromycin and the remaining 18 received sequential therapy. Seventy-two patients consumed at least 90% of the capsules, while only one did not complete the therapy due to adverse events (nausea and diarrhea). By ITT analysis, BQT was successful in 62 subjects (eradication rate 84.9%, 95%CI 76.7–93.1%). By PP analysis, the eradication rate was 86.1% (95%CI 78.1–94.1%).Adverse events were observed in 14 subjects (20.5%).In conclusion, our report confirmed that BQT is effective as an empiric second-line regimen.

A Validated HPTLC Densitometric Method for The Quantitative Determination of Ubidecarenone in Bulk and in Capsule Formulation

A new, simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and validated for the estimation of ubidecarenone in bulk and in capsule formulation. The chromatographic separation was performed on aluminium TLC plates precoated with silica gel 60F254 as a stationary phase and methanol:water (7:3) as a mobile phase. Detection was performed densitometrically in the absorbance mode at 280nm for the evaluation of chromatograms. The system has given well sharp peak of ubidecarenone (Rf=0.51±0.02). The linearity of the method was established in the range of 1-6 ng/µL with correlation coefficient (r2) of 0.9995. The method was validated for precision, accuracy, robustness, ruggedness, LOD, and LOQ as per ICH guidelines. The limit of detection was found to be 0.0392 ng/µL, whereas the limit of quantitation was found to be 0.1189 ng/µL. The percentage label claim for ubidecarenone in the capsule formulation was found to be 99.96±0.4703. The accuracy of the method was confirmed by recovery studies. The percentage recovery was found to be in the range of 100.10-101.45% for ubidecarenone. The % RSD value was found to be less than 2. The low %RSD value indicates that there is no interference due to excipients used in the formulation. Hence, the developed method was found to be simple, precise, accurate, and rapid for the analysis of ubidecarenone in bulk and pharmaceutical formulation and it can be effectively applied for the quality control analysis of ubidecarenone in bulk and pharmaceutical formulation.

Capsule-C: an improved Steinernema carpocapsae capsule formulation for controlling Agrotis ipsilon Hufnagel (Lepidoptera: Noctuidae)

Background Entomopathogenic nematodes (EPNs) have long been used for controlling soil-dwelling insects. Steinernema carpocapsae HB310, previously showed a high virulence against many pests including Agrotis ipsilon Hufnagel (Lepidoptera: Noctuidae). Due to the lack of durable formulations, up until now, S. carpocapsae HB310 has thus far been prevented from use in large-scale farming. The present study aimed to get a better EPNs capsule formulation suitable for long-term storage and effective application. Results An improved EPNs capsule formulation, herein named: Capsule-C was prepared by the following composition: Solution I: 18% glycerol, 0.075% formaldehyde, 1% sodium alginate, 0.2% xanthan gum, 0.5% potassium sorbate, 9% glucose, 2% fructose, 2% sucrose, and the remainder was distilled water. The nematodes suspension was added to the alginate mixture in 2 × 104 IJs/mL; Solution II: 18% glycerol, 0.075% formaldehyde, 0.5% calcium chloride, 0.5% potassium sorbate, with the remainder being distilled water. After storage for 180 days at 16 °C and 100% RH, the survival rate of nematodes in Capsule-C was 75.68 ± 0.48% and the nematodes caused 82.33 ± 1.45% mortality in the 5th instar larvae of Galleria mellonella. A. ipsilon larvae preferred to chew and ingest Capsule-C due to the addition of the glucose compound. The feeding rate of A. ipsilon larvae on Capsule-C reached to 100% within 24 h and the larval mortality of A. ipsilon was 90.48 ± 6.35%. Conclusion EPNs-containing capsules were as effective as sprayed EPNs in water solution at killing A. ipsilon. These results will provide ideas to acquire a stable and efficient EPNs capsule formulation and further promote the application of environmental friendly biological pesticides.

DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR SIMULTANEOUS ESTIMATION OF CLIDINIUM BROMIDE, RABEPRAZOLE, CHLORDIAZEPOXIDE AND DICYCLOMINE HYDROCHLORIDE

Clidinium bromide, rabeprazole sodium, chlordiazepoxide and dicyclomine hydrochloride(COLIWIN-R) drug combinations are used for the treatment of gastric acidity, anxiety, intestinal ulcers, abdominal cramps, irritable bowel syndrome and abdominal pain. A high performance liquid chromatographic method has been developed and validated for the simultaneous determination of clidinium bromide, rabeprazole sodium, chlordiazepoxide and dicyclomine hydrochloride in capsules dosage forms using WATERS C18 column (50 mm × 4.6 mm, 5 µm) with mobile phase consisting of methanol, acetonitrile and phosphate buffer 40:30:30 (V/V) (0.05M, pH 4.0 adjusting with 0.5% ortho phosphoric acid) at a flow rate of 1.0 mL/ min monitoring the effluents at 220 nm. The retention times of clidinium bromide, rabeprazole sodium, chlordiazepoxide and dicyclomine hydrochloride in capsule formulation were found to be 2.9 min, 3.5 min, 4.7 min, and 8.0 min, respectively. The method was validated according to the ICH guidelines for specificity, LOD, LOQ, precision, accuracy, linearity, ruggedness and robustness. The method show good reproducibility and recovery with % RSD less than 2. The proposed method was found to be simple, specific, precise, accurate and linear. Hence, it can be applied for routine analysis of clidinium bromide, rabeprazole sodium, chlordiazepoxide and dicyclomine hydrochloride in pharmaceutical combined dosage forms.

Capsule Formulation Essay of Herbal Extracts of Trunk Bark of Anogeissus leiocarpus (DC) Guill. Et Perr. (Combretaceae) for the Treatment of Hypertension

Introduction: The trunks barks of Anogeissus leiocarpus contains multiple antihypertensive components and are, therefore, widely used for the treatment of hypertension. Objective: This study aimed to formulate capsules containing a freeze-dried aqueous extract of the trunk’s barks. Methodology: Three (03) capsule formulations were prepared using wet granulation from lyophilised aqueous extract of Anogeissus leiocarpus trunk bark, with different proportions of excipients including Corn starch used as the diluent, Polyvinylpyrrolidone K25 (PVP) used as binding agent and Magnesium stearate used as gliding agents. The filling of capsules was done using a semi-automatic capsule filler with empty capsules of size 3. The flow properties of the granules and the quality control were performed according to the European Pharmacopoeia 10th ed. Results/Discussion: All granules had good flow properties, and the F1 and F3 formulations had the best pharmaceutics characteristics according to the recommendations of the European Pharmacopoeia 10th ed. The mean levels of phenolic tracers were 0.039±0.0097 mg gallic acid equivalent per capsule (GAE/capsule) for the F1 formulation and 0.059±0.0063 mg GAE/capsule for the F3 formulation. Conclusion: This study allowed the galenic formulation of capsules based on extracts of good characteristic quality for the treatment of hypertension. Keywords: Anogeissus leiocarpus, extract, excipients, formulations, capsule

Separation and Simultaneous Quantitation of Tolterodine Tartrate and Tamsulosin Hydrochloride in Capsule Formulation by using Stability-Indicating RP-HPLC-DAD Method

In the present work, a precise, accurate and selective stability-indicating RP-HPLC method has been developed and validated according to International Conference on Harmonization guidelines Q2-(R1) for the simultaneous quantitative determination of Tolterodine tartrate and Tamsulosin hydrochloride in bulk and its capsule formulation. The chromatographic separation was achieved on Hypersil octadecyl silane C18 (250 x 4.6mm, 5 μm) column at room temperature and mobile phase comprised of methanol: 0.05 M phosphate buffer, pH 7.0 in the ratio of 90:10 V/V. The flow rate of mobile phase was set at 1.0mL/min and compounds were monitored at 255nm using photodiode array detector. Tamsulosin hydrochloride and Tolterodine tartrate retention time were found to be 4.15±0.2 min and 8.42±0.2 min, respectively. The drug substances and products were subjected to acid hydrolysis, alkali hydrolysis, oxidative hydrolysis, photolytic and thermal degradation. The percent degradation of drugs was calculated in all stressed conditions. The analytical method validation parameters such as linearity, accuracy, precision, detection limits, quantitation limits and robustness indicate that drug substances and products were efficiently separated in present of their degradants and successfully applied for the routine analysis of Tolterodine tartrate and Tamsulosin hydrochloride in bulk and its capsule formulation in the quality control laboratory.

LIPOPHILICITY, AQUEOUS SOLUBILITY, AND DEGREE OF IONIZATION OF ATRACTYLODIN AND β-EUDESMOL, THE BIOACTIVE COMPOUNDS ISOLATED FROM ATRACTYLODES LANCEA

Objective: The study aimed to evaluate the critical physicochemical properties (lipophilicity, aqueous solubility, and degree of ionization) of atractylodin and β-eudesmol using in vitro testing. Methods: Lipophilicity (Log P and Log D) was determined using the shake-flask method (n-octanol/water partition). Aqueous solubility was determined using kinetic solubility assay in media with pH ranging from 1.2 to 7.4. The degree of ionization (pKa) was determined using the potentiometric titration method. Results: Log P and Log D values of 3.0-5.0 suggested moderate lipophilicity of both compounds. Both exhibited low aqueous solubility over the investigated pH range (0.08-0.93 and 1.97-32.48 μg/ml for atractylodin and β-eudesmol, respectively). Based on the pKa values of 9.63 (atractylodin) and 9.12 (b-eudesmol), both are classified as basidic compounds. Conclusion: Atractylodin and β-eudesmol are classified as BCS class II drugs. The physicochemical parameters of both compounds obtained from the current study will be further applied for in silico prediction of their ADME (absorption, distribution, metabolism, and excretion) properties. In addition, PBPK modeling will be used for the prediction of optimal dose regimens of the capsule formulation of the standardized extract of Atractylodes lancea for first-in-human (FIH) and phase II studies in patients with cholangiocarcinoma.

Pharmacokinetics and Safety of an Improved Oral Formulation of Paltusotine, a Selective, Non-Peptide Somatostatin Receptor 2 (SST2) Agonist for the Treatment of Acromegaly

Depot injection formulations of peptide somatostatin receptor ligands (SRLs) are routinely used to treat acromegaly and neuroendocrine tumors (NETs). Paltusotine (CRN00808), an orally administered small molecule nonpeptide selective somatostatin receptor 2 (SST2) agonist has been shown to maintain GH and IGF-1 levels in acromegaly patients previously on depot SRLs (ACROBAT Edge NCT03789656). In this study, a capsule formulation was used, which did not exhibit dose proportional pharmacokinetics (PK) at doses >40 mg, required a 2-hour post-dose fast in overnight fasted patients, and had the potential for reduced bioavailability when taken with proton-pump inhibitors (PPI). A spray-dried dispersion (SDD) tablet formulation was developed with improved solubility in the physiological pH range and its performance was evaluated in healthy volunteers. Male and female healthy volunteers who met inclusion/exclusion criteria were enrolled in a single-center Phase 1 study (ANZCTR registration ACTRN12619001562167). A Cohort of 12 subjects was administered a single dose of paltusotine in a four-period cross-over design. Periods 1 and 2 assessed the effect of lansoprazole (a PPI) on PK of 20 mg dose of paltusotine SDD tablets. In Period 3, 20 mg dose of paltusotine SDD tablets was co-administered with a high fat, high-calorie meal. In Period 4, a 60 mg dose of paltusotine SDD tablets was administered to assess dose proportionality. In a separate cohort of subjects (n=12; also, a 4-period cross-over design), the relative bioavailability of capsules and SDD tablets was assessed, and the effect of food administration 0.5, 1, and 2 hour post-dose was evaluated. Subsequently, in another cohort of 12 subjects (a 3-period cross-over design), dose proportionality of the SDD tablets was evaluated at 40 mg and 80 mg dose with a 1-hour post-dose fast. A 4 hours post-dose fast was also assessed for the 80 mg dose. Pharmacokinetics and safety of paltusotine were evaluated. Paltusotine was generally well tolerated in this study. SDD tablets exhibited dose proportional increase in total systemic exposure (AUC) up to a dose of 80 mg. Healthy volunteers pretreated with the PPI, lansoprazole (15 mg bid for 3 days), and co-administered with paltusotine SDD tablets exhibited a small decrease (approximately 25%) in systemic exposure to paltusotine compared with the same subjects that had washed out from the PPI-pretreatment. SDD tablets exhibited significant reduction in systemic exposure when co-administered with a high-fat, high-calorie meal. However, the SDD formulation was less sensitive to timing of post-dose food administration compared to the capsule formulation. These data suggest that the SDD tablet formulation of paltusotine improves flexibility in dose administration, can be co-administered with PPIs and other agents that increase stomach pH, and reduces the post-dose fasting requirement.

Pharmacokinetics and Safety of a Novel Oral Liquid Formulation of 13-cis Retinoic Acid in Children with Neuroblastoma: A Randomized Crossover Clinical Trial

(1) Background: 13-cis-retinoic acid (13-CRA) is a key component of neuroblastoma treatment protocols. This randomized crossover study compares the pharmacokinetics (PK), safety and palatability of a novel oral liquid formulation to the current method of extracting 13-CRA from capsules. (2) Methods: Pharmacokinetics was evaluated in two consecutive treatment cycles. Patients were randomized to receive either liquid or capsule formulation on cycle 1 and then crossed over to the alternative formulation on cycle 2. The daily dose was 200 mg/m2, reduced to 160 mg/m2 in patients with weight ≤ 12 kg. (3) Results: A total of 20 children, median (range) age 4.3 (1–11.6) y were recruited. Pharmacokinetic data were pooled and a population model describing the disposition of 13-CRA and 4-oxo-13-CRA was developed. Bioavailability of the liquid formulation was estimated to be 65% higher (95% CI; 51–79%) than the extracted capsule. CmaxSS and AUC(0-12)SS estimates were also significantly higher; mean (95% CI) differences were 489 (144–835) ng/mL and 3933 (2020–5846) ng/mL·h, respectively (p < 0.01). There were no significant differences in reported adverse effects. Parents found dosing considerably easier with liquid formulation. (4) Conclusions: The pharmacokinetics, safety and palatability of a new liquid formulation of 13-CRA compares favorably to 13-CRA extracted from capsules. Clinical Trial Registration: clinicaltrial.gov NCT03291080.

Effects of Sources or Formulations of Vitamin K3 on Its Stability during Extrusion or Pelleting in Swine Feed

Two studies were conducted to determine the stability of vitamin K3 (VK3) in swine diets during extrusion or pelleting. The two sources were menadione sodium bisulfite (MSB) and menadione nicotinamide bisulfite (MNB), and the three formulations were crystal micro-capsule formulation and micro-sphere formulation. The recovery of six types of VK3 in swine diets was investigated after extrusion at 100 °C or 135 °C in Experiment 1. The recovery of six types of VK3 was investigated when the diets were pelleted at 60 °C (low temperature; LT) or 80 °C (high temperature; HT) and the length to diameter ratios were 5.2:1 (low length to diameter ratio; LR) or 7.2:1 (high length to diameter ratio; HR) in Experiment 2. In Experiment 1, MNB recovery (72.74%) was higher than MSB recovery (64.67%) after extrusion, while recovery of VK3 of crystal (74.16%) was higher than the recovery of micro-capsule (65.25%) and micro-sphere (66.72%). The recovery of VK3 (70.88%) was higher when extruded at 100 °C than that at 135 °C (66.54%). In Experiment 2, MNB recovery (86.21%) was higher than MSB recovery (75.49%) after pelleting, while the recovery of VK3 of micro-capsule (85.06%) was higher than the recovery of crystal (81.40%) and micro-sphere (76.09%). The recovery of VK3 (75.50%) was lower after HTHR pelleting than LTLR (83.62%), LTHR (81.52%) or HTLR (82.76%) treatment. Our results show that MNB has greater stability than MSB. VK3 of crystal or VK3 of micro-capsule were recommended for extrusion or pelleting, respectively.