Behavioural Parameters of Circadian Rhythm Are Not Correlated with Dim Light Melatonin Onset: An Observational Study on Healthy Volunteers.

Dim light melatonin onset (DLMO) is considered the most reliable marker of the circadian rhythm phase in humans. DLMO may moderately correlate with sleep onset and sleep offset time. There are no sufficient data about the correlations between DLMO and clinical scales assessing sleep quality and daytime symptoms of poor night sleep. The aim of the study was to determine the association between DLMO and basic sleep parameters from actigraphy and sleep diaries, as well as the association between DLMO and the following insomnia clinical scales: the Athens Insomnia Scale (AIS), Insomnia Severity Index (ISI), Epworth Sleepiness Scale (ESS), and chronotype questionnaires: Morningness-Eveningness Questionnaire (MEQ) and Composite Scale of Morningness (CSM). Participants of the study were healthy volunteers. Sleep parameters were measured by sleep diaries and actigraphy, and the following clinical scales: the AIS, ISI, and ESS, and chronotype questionnaires: MEQ and CSM. DLMO was calculated based on plasma melatonin concentration. The blood samples were collected hourly at five time points between 20:00 and 00:00 during the session in dim red light (<50 lux). Melatonin concertation was determined by LC-MS/MS. Twenty-one volunteers participated in the study. DLMO was calculated in 12 participants. There was a significant correlation between DLMO and ISI (r = 0.60, p = 0.038) and ESS (r = 0.61, p = 0.034). The correlation coefficient between the DLMO and the AIS was also high, however insignificant (r = 0.57, p = 0.054). There were no significant correlations between DLMO and chronotype scales MEQ and CSM. DLMO did not correlate with sleep onset and sleep offset; however, DLMO correlated with the Sleep Fragmentation Index (SFI) (r = 0.67, p = 0.017). DLMO is associated with poorer sleep maintenance, a stronger feeling of insomnia, and sleepiness during the day. Simultaneously, chronotype pattern and circadian rhythm parameters do not correlate with DLMO. Biological circadian rhythm does not reflect the real-life sleep-wake rhythm, indicating that the lifestyle is more often disconnected from the biological clock.

Dispensing of minitablets - Has the problem been resolved?

Recently, minitablets have been given extensive coverage in literature, as they are perfectly matched to the current therapy individualization trend. Within this scope, special attention is paid to minitablets that enable convenient drug intake for patients with swallowing problem. However, the packaging system, dispensing the necessary amount of drug units and safe administration still remain unsolved problems or are partially overlooked. Although there are many different approaches towards dosing tablets, only a few seem to be tailored to particularly small tablets. Moreover, none of these approaches meets all the user's expectations. This paper comprehensively elaborates and critically discusses the available dosing options like sachets, blisters, home electronic dispensing systems and minitablets manual dispensers. Additional tests have been also conducted to simulate the handling and dosing procedure with 2 mm diameter placebo minitablets. Despite many advantageous inventions, it has been revealed that further efforts are necessary to identify the optimal design that would allow to eliminate the shaking procedure, adjust cavities diameter or provide better protection against humidity. Nevertheless, the current trend may lead to individual therapy becoming more convenient, safe and reliable, especially in pediatric and geriatric patients.

Assessment of dim light melatonin onset based on plasma and saliva samples.

Melatonin (MELA) is a nocturnal hormone involved in the regulation of the circadian rhythm. MELA can be detected in plasma and saliva, and its salivary concentration strongly correlates with its plasma concentration. Dim light melatonin onset (DLMO) is considered to be the most accurate objective marker for assessing the circadian phase. The purpose of the study was to establish a method for the determination of MELA in plasma and saliva based on the liquid chromatography with tandem mass spectrometry (LC-MS/MS) and compare DLMO using both plasma and saliva matrices. The validation of the LC-MS/MS methods was performed in accordance with the European Medicines Agency (EMA) guideline. The study was conducted on a group of 21 volunteers, male and females, aged 26-54 years. Plasma and saliva were collected at five time points: between 20:00 and 00:00 hours. The MELA concentration was determined by the LC-MS/MS. The DLMO was considered as the point in time when MELA concentration exceeds 20 pg/mL in plasma and 7 pg/mL in saliva. The correlation coefficient between the plasma and salivary MELA concentration was r = 0.764 (p < .001). The ratio of the plasma/saliva MELA concentrations was 2.87. The mean time of the DLMO in the plasma was 21:30 ± 0:45 hours, and in the saliva was as follows: 21:34 ± 1:00 hours. The correlation between the DLMO, calculated based on the plasma and saliva MELA profiles, was r = 0.679 (p < .05). The determination of salivary MELA concentration using LC-MS/MS allows for the determination of the DLMO. Our method may be applied in clinical practice for the diagnosis and monitoring of circadian rhythm disorders.Abbreviations: CE: Collision Energy; CID: Collision-Induced Dissociation; DL: Desolvation Module; DLMO: Dim Light Melatonin Onset; EFSA: European Food Safety Authority; EMA: European Medicines Agency; ESI: electrospray ionization; HB: heat block; HPLC: high performance liquid chromatography; IS: internal standard; K3EDTA: ethylenediaminetetraacetic acid tripotassium salt; LC-MS/MS: liquid chromatography with tandem mass spectrometry; LLE: liquid-liquid extraction; LLOQ: lower limit of quantification; MELA: melatonin; MELA-D4: melatonin-d4; MRM: multiple reaction monitoring; Q1: quadrupole 1; Q3: quadrupole 3; RE: relative error; RIA: radioimmunoassay; RSD: relative standard deviation; SD: standard deviation; ULOQ: upper limit of quantification.

Implementing the Design of Experiments (DoE) Concept into the Development Phase of Orodispersible Minitablets (ODMTs) Containing Melatonin.

Development of orodispersible minitablets (ODMTs) requires consideration of aspects related to small dimensions, while ensuring short disintegration time with sufficient mechanical stability. In order to meet these and other critical quality attributes (CQAs), quality by design is encouraged. According to this approach, formulation and compression process factors were systematically studied using design of experiments (Plackett-Burman for screening purposes, full and fractional factorial design for in-depth characterization) to understand their influence on CQAs of orodispersible minitablets containing melatonin. Mathematical models describing the relationships between processing variables and attributes such as resistance to crushing and disintegration time were successfully developed, characterized by high coefficients of determination (R2adj = 0.90-0.97) and prediction errors in the range (+2.4 to -10.8%). In conclusion, based on these models, the design space was created for melatonin ODMTs, ensuring the product's quality and process robustness. Moreover, the study demonstrated the suitability of texture analysis as an alternative to compendial measurement methods of resistance to crushing and disintegration time. Graphical abstract.

Biorelevant In Vitro Release Testing and In Vivo Study of Extended-Release Niacin Hydrophilic Matrix Tablets.

Niacin (nicotinic acid, NA) is administered orally as an antihyperlipidemic agent in extended-release (ER) tablets in high doses. Due to rapid absorption and extensive metabolism (non-linear pharmacokinetics), the drug plasma levels are highly variable, which may correlate with side effects. Interestingly, this erratic drug delivery behavior of niacin ER products cannot be clarified by compendial in vitro release testing. The standard dissolution tests do not allow to mimic the selected GI tract characteristics in order to estimate the robustness of formulation under the variability of the physiological conditions. These are characterized by the pH value, impact of motility forces and composition, as well as volume of GI liquids. Our paper demonstrates a comparison of a newly developed ER HPMC niacin formulation with an originator product. The research aimed to design a robust matrix tablet of comparable biopharmaceutical behavior, safety and efficacy. The extensive in vitro investigation, including dynamic studies in flow-through cell apparatus and stress test device, forms the basis for the evaluation of nicotinic acid plasma concentrations in vivo. The occurrence of erratic, multiple NA plasma peaks after the administration of both extended-release products is a result of its local input excess over the metabolic threshold (at the level corresponding to maximum 2% of the administered dose, i.e., 20 mg of drug) due to the mechanical stresses of physiological intensity. We demonstrate how this behavior is similar for both marketed and test products. In this context, we describe how a robust ER matrix and well-designed formulation does not guarantee the test product's bioequivalence to the comparator one out of reasons unrelated to technology and biopharmaceutical properties, but because of the active compound's intrinsic pharmacokinetic characteristics, i.e., highly variable, extensive metabolism of nicotinic acid.

Two Aspergillus nidulans genes encoding methylenetetrahydrofolate reductases are up-regulated by homocysteine.

Methylenetetrahydrofolate reductase (MTHFR) catalyzes the reduction of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, a co-substrate in the synthesis of methionine from homocysteine. We have cloned and characterized two Aspergillus nidulans genes encoding MTHFRs: metA and metF. Mutations in either gene result in methionine requirement; the metA-encoded enzyme is responsible for only 10-15% of total MTHFR activity. These two enzymes belong to different classes of MTHFRs. Mutations in metA but not in the metF gene are suppressed by mutations resulting in enhancement of homocysteine synthesis. The expression of both genes is up-regulated by homocysteine.