Small-scale in vitro methods for evaluating the impact of gastrointestinal transfer on luminal drug / drug product performance in the fasted state

2020 ◽  
Author(s):  
Πάτρικ Ο' Ντουάιρ
Keyword(s):  
Drug Product ◽  
Product Performance ◽  
Small Scale ◽  
In Vitro Methods ◽  
Drug Products ◽  
Fasted State ◽  
The Impact

Η βιοδιαθεσιμότητα ενός φαρμάκου μετά από per os χορήγηση επηρεάζεται από πολλές διαδικασίες. Η γαστρεντερική μεταφορά αποτελεί μια σημαντική φυσιολογική διαδικασία, που επηρεάζει της συγκεντρώσεις του φαρμάκου στο λεπτό έντερο, δηλαδή στην περιοχή από την οποία απορρόφουνται τις περισσότερες φορές τα per os χορηγούμενα φάρμακα. Στην παρούσα εργασία αναπτύχθηκαν δυο μικρής κλίμακας μέθοδοι για την εκτίμηση της σημασίας της γαστρεντερικής μεταφοράς στην απορρόφηση ενός φαρμάκου μετά από per os χορήγηση κατά τη διάρκεια της διαπεπτικής περιόδου. Οι μέθοδοι βασίστηκαν σε ένα διφασικό σύστημα διάλυσης δύο σταδίων και σε μια συσκευή διάλυσης-διαπέρασης δυο σταδίων. Η διφασική μέθοδος διάλυσης προσομοιώνει τη διαδικασία της μεταφοράς μέσω του εντερικού επιθηλίου με τη χρήση μιας στοιβάδας δεκανόλης. Η μέθοδος διάλυσης-διαπέρασης προσομοιώνει τη διαδικασία μεταφοράς μέσω του γαστρεντερικού επιθηλίου με τη χρήση μιας συνθετικής λιπιδικής μεμβράνης και ενός διαμερίσματος υποδοχής του φαρμάκου. Οι δυο μικρής κλίμακας μέθοδοι δυο σταδίων αναπτύχθηκαν και αξιολογήθηκαν με βάση δεδομένα δυο ασθενών βάσεων που ανήκουν στην Τάξη ΙΙ της Βιοφαρμακευτικής Ταξινόμησης των Φαρμάκων, της διπυριδαμόλης και της κετοκοναζόλης. Οι ρυθμοί καθίζησης, που υπολογίσθηκαν από τα δεδομένα των πειραμάτων με τις δυο μεθόδους, χρησιμοποιήθηκαν για την προσομοίωση των απεικονίσεων των συγκεντρώσεων στο πλάσμα μετά από χορήγηση σε ενήλικες με τη βοήθεια φυσιολογικών φαρμακοκινητικών μοντέλων. Οι τιμές των φαρμακοκινητικών παραμέτρων, που υπολογίσθηκαν από τις απεικονίσεις, συγκρίθηκαν με τις τιμές των φαρμακοκινητικών παραμέτρων, που είχαν υπολογισθεί από προηγούμενες κλινικές μελέτες των φαρμάκων σε υγιείς ενήλικες. Τα δεδομένα της διφασικής μεθόδου διάλυσης οδήγησαν σε καλύτερη εκτίμηση των φαρμακοκινητικών παραμέτρων των δυο ασθενών βάσεων. Τα δεδομένα της μεθόδου διάλυσης-διαπέρασης οδήγησαν σε υπερεκτίμηση της καθίζησης, πιθανότατα λόγω της αργής ροής μεταφοράς στο διαμέρισμα υποδοχής. Στη συνέχεια, οι δυο μικρής κλίμακας μέθοδοι δυο σταδίων αξιολογήθηκαν χρησιμοποιώντας φαρμακευτικά προϊόντα με αυξημένες δυνατότητες (enabling drug products) που περιλάμβαναν άμορφες στερεές διασπορές, ένα σύμπλοκο φαρμάκου με κυκλοδεξτρίνη και λιπιδικά προϊόντα. Για τη μελέτη των λιπιδικών προϊόντων η διφασική μέθοδος τροποποιήθηκε για να ενσωματωθεί και η διαδικασία της πέψης των εκδόχων. Συνολικά, η διφασική μέθοδος ήταν περισότερο χρήσιμη από τη μέθοδο-διάλυσης διαπέρασης και τα δεδομένα σχετίστηκαν καλύτερα με τα in vivo δεδομένα. Τέλος, η χρησιμότητα των δυο μικρής κλίμακας μεθόδων δυο σταδίων σε σχέση με δυο άλλες χρησιμοποιούμενες μεθόδους, τη διατάξη μικρού περιστρεφόμενου πτερυγίου Erweka και τη διάταξη ΒioGIT, μελετήθηκε με βάση δεδομένα δυο φαρμάκων που ανήκουν στην Τάξη ΙΙ της Βιοφαρμακευτικής Ταξινόμησης των Φαρμάκων, της δικλοφενάκης (ασθενές εξύ) και της ριτοναβίρης (ασθενής βάση). Με όλες τις μεθόδους, η δικλοφενάκη διαλύθηκε πολύ γρήγορα σε μέσα που προσομοιώνουν σε Επίπεδο ΙΙ τα περιεχόμενα του λεπτού εντέρου, τόσο μετά από μορφοποίηση σε σκόνη όσο και μετά από μορφοποίηση σε δισκίο. Φυσιολογική φαρμακοκινητική μοντελοποίηση των δεδομένων έδειξε, ότι μετά από χορήγηση προϊόντων άμεσης αποδέσμευσης δικλοφενάκης, η απορρόφηση εξαρτάται από τη γαστρική κένωση και τη μεταφορά μέσω του εντερικού επιθηλίου, ανεξάρτητα από τον τύπο του προϊόντος. Αντίθετα, η άμορφη στερεή διασπορά της ριτοναβίρης οδηγεί σε καθίζηση του φαρμάκου στον αυλό του λεπτού εντέρου. Όλες οι μέθοδοι που προσομοίωσαν την αλλαγή των συνθηκών από το στόμαχο στο λεπτό έντερο προσομοίωσαν επαρκώς την επίδραση του μειωμένου ρυθμού έκκρισης γαστρικού οξέος στη συμπεριφορά του προϊόντος in vivo. Με βάση τα δεδομένα συνολικά, κάθε μέθοδος παρουσιάζει ιδιαίτερα προτερήματα και αδυναμίες. Συνολικά, με βάση τα αποτελέσματα της παρούσας μελέτης η διφασική μέθοδος θα μπορούσε να διευκολύνει σημαντικά στα αρχικά στάδια της ανάπτυξης νέων φαρμάκων, ιδιαίτερα αν συνδυαστεί με φυσιολογικά φαρμακοκινητικά μοντέλα.

scholarly journals In Vivo Predictive Dissolution Testing of Montelukast Sodium Formulations Administered with Drinks and Soft Foods to Infants

2020 ◽  
Vol 21 (7) ◽  
Author(s):  
J. Martir ◽  
T. Flanagan ◽  
J. Mann ◽  
N. Fotaki
Keyword(s):  
Drug Product ◽  
Drug Dissolution ◽  
Simulated Gastric Fluid ◽  
Dissolution Testing ◽  
Fed State ◽  
Fasted State ◽  
Direct Administration ◽  
The Impact

Abstract In vitro dissolution testing conditions that reflect and predict in vivo drug product performance are advantageous, especially for the development of paediatric medicines, as clinical testing in this population is hindered by ethical and technical considerations. The aim of this study was to develop an in vivo predictive dissolution test in order to investigate the impact of medicine co-administration with soft food and drinks on the dissolution performance of a poorly soluble compound. Relevant in vitro dissolution conditions simulating the in vivo gastrointestinal environment of infants were used to establish in vitro-in vivo relationships with corresponding in vivo data. Dissolution studies of montelukast formulations were conducted with mini-paddle apparatus on a two-stage approach: infant fasted-state simulated gastric fluid (Pi-FaSSGF; for 1 h) followed by either infant fasted-state or infant fed-state simulated intestinal fluid (FaSSIF-V2 or Pi-FeSSIF, respectively; for 3 h). The dosing scenarios tested reflected in vivo paediatric administration practices: (i.) direct administration of formulation; (ii.) formulation co-administered with vehicles (formula, milk or applesauce). Drug dissolution was significantly affected by co-administration of the formulation with vehicles compared with after direct administration of the formulation. Montelukast dissolution from the granules was significantly higher under fed-state simulated intestinal conditions in comparison with the fasted state and was predictive of the in vivo performance when the granules are co-administered with milk. This study supports the potential utility of the in vitro biorelevant dissolution approach proposed to predict in vivo formulation performance after co-administration with vehicles, in the paediatric population.

scholarly journals Evaluation of Excipient Risk in BCS Class I and III Biowaivers

2022 ◽  
Vol 24 (1) ◽  
Author(s):  
Melissa Metry ◽  
James E. Polli
Keyword(s):  
Drug Product ◽  
Scale Up ◽  
Drug Formulation ◽  
Product Regulatory ◽  
Drug Bioavailability ◽  
Drug Products ◽  
Fda Guidance ◽  
Regulatory Guidance

AbstractThe objective of this review article is to summarize literature data pertinent to potential excipient effects on intestinal drug permeability and transit. Despite the use of excipients in drug products for decades, considerable research efforts have been directed towards evaluating their potential effects on drug bioavailability. Potential excipient concerns stem from drug formulation changes (e.g., scale-up and post-approval changes, development of a new generic product). Regulatory agencies have established in vivo bioequivalence standards and, as a result, may waive the in vivo requirement, known as a biowaiver, for some oral products. Biowaiver acceptance criteria are based on the in vitro characterization of the drug substance and drug product using the Biopharmaceutics Classification System (BCS). Various regulatory guidance documents have been issued regarding BCS-based biowaivers, such that the current FDA guidance is more restrictive than prior guidance, specifically about excipient risk. In particular, sugar alcohols have been identified as potential absorption-modifying excipients. These biowaivers and excipient risks are discussed here.

scholarly journals Analysis of Non-Pivotal Bioequivalence Studies Submitted in Abbreviated New Drug Submissions for Delayed-Release Drug Products

2017 ◽  
Vol 20 ◽  
pp. 252
Author(s):  
Paramjeet Kaur ◽  
Xiaojian Jiang ◽  
Ethan Stier
Keyword(s):  
Coating Layer ◽  
Drug Product ◽  
Enteric Coating ◽  
Drug Products ◽  
New Drug ◽  
Delayed Release ◽  
The Us ◽  
Generic Products ◽  
The Impact

The US FDA’s rule on “Requirements for Submission of Bioequivalence Data” requiring submission of all bioequivalence (BE) studies conducted on the same formulation of the drug product submitted for approval was published in Federal Register in January 2009. With the publication of this rule, we evaluated the impact of data from non-pivotal BE studies in assessing BE and identified the reasons for failed in vivo BE studies for generic oral delayed-release (DR) drug products only. We searched the Agency databases from January 2009 toDecember 2016 to identify Abbreviated New Drug Applications (ANDAs) submitted for DR drug products containing non-pivotal BE studies. Out of 202 ANDAs, 43 ANDAs contained 102 non-pivotal BE studies. Forty-nine non-pivotal BE studies were conducted on the to-be-marketed (TBM) formulation and 53 were conducted on formulations different from the TBM formulation. These experimental formulations primarily differed in the ratio of components of the enteric coating layer and/or amount (i.e., %w/w) of enteric coating layer. Of the 49 non-pivotal BE studies conducted on the TBM formulation, 41 failed to meet the BE acceptance criteria. The majority of failed non-pivotal BE studies on the TBM DR generic products had insufficient power, which was expected as these studies are exploratory in nature and not designed to have adequate power to pass the BE statistical criteria. In addition, among the failed non-pivotal BE studies on the TBM DR generic products, the most commonly failing pharmacokinetic parameter was Cmax. The data from these non-pivotal BE studies indicate that inadequate BE study design can lead to failure of the BE on the same formulation. Also, the non-pivotal BE studies on formulations different from the TBM formulation help us link the formulation design to the product performance in vivo. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

Analysis of antidepressant properties of some Papaver species by in vivo and in vitro methods

Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
OML Bayazeid ◽  
F Yalcin ◽  
M İlhan ◽  
H Karahan ◽  
E Kupeli-Akkol ◽  
...  
Keyword(s):  
In Vitro Methods

The Effects of Some Synthetic Compounds on In Vitro Fibrinolytic Activity Measured by Different Methods and the Relevance to Activity In Vivo

1972 ◽  
Vol 28 (03) ◽  
pp. 351-358
Author(s):  
A.J Baillie ◽  
A. K Sim
Keyword(s):  
Inhibitory Activity ◽  
Substrate Concentration ◽  
Fibrinolytic Activity ◽  
Synthetic Compounds ◽  
In Vitro Methods ◽  
Narrow Concentration Range

SummaryThe activity of several synthetic compounds, rated from good to poor (or inactive) fibrinolytic activators, has been assessed by two different commonly-used in vitro methods. Compounds shown to be active over a narrow concentration range in the hanging clot test were shown to be inhibitors of plasmin and trypsin in the casein-olytic test. The inhibitory activity of these compounds was shown to increase with increasing substrate concentration and apparent activity in the hanging clot test. Possible explanations and relevance of these observations are discussed.

The impact of Zataria multiflora Boiss extract on in vitro and in vivo Th1/Th2 cytokine (IFN-γ/IL4) balance

2013 ◽  
Vol 150 (3) ◽  
pp. 1024-1031 ◽  
Author(s):  
Mohammad Hossein Boskabady ◽  
Sakine Shahmohammadi Mehrjardi ◽  
Abadorrahim Rezaee ◽  
Houshang Rafatpanah ◽  
Sediqeh Jalali
Keyword(s):  
Zataria Multiflora ◽  
Th2 Cytokine ◽  
Ifn Γ ◽  
The Impact

scholarly journals The impact of FGF19/FGFR4 signaling inhibition in antitumor activity of multi-kinase inhibitors in hepatocellular carcinoma

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hiroaki Kanzaki ◽  
Tetsuhiro Chiba ◽  
Junjie Ao ◽  
Keisuke Koroki ◽  
Kengo Kanayama ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Molecular Mechanisms ◽  
Kinase Inhibitors ◽  
Progression Free Survival ◽  
Erk Signaling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antitumor Effects ◽  
The Impact

AbstractFGF19/FGFR4 autocrine signaling is one of the main targets for multi-kinase inhibitors (MKIs). However, the molecular mechanisms underlying FGF19/FGFR4 signaling in the antitumor effects to MKIs in hepatocellular carcinoma (HCC) remain unclear. In this study, the impact of FGFR4/ERK signaling inhibition on HCC following MKI treatment was analyzed in vitro and in vivo assays. Serum FGF19 in HCC patients treated using MKIs, such as sorafenib (n = 173) and lenvatinib (n = 40), was measured by enzyme-linked immunosorbent assay. Lenvatinib strongly inhibited the phosphorylation of FRS2 and ERK, the downstream signaling molecules of FGFR4, compared with sorafenib and regorafenib. Additional use of a selective FGFR4 inhibitor with sorafenib further suppressed FGFR4/ERK signaling and synergistically inhibited HCC cell growth in culture and xenograft subcutaneous tumors. Although serum FGF19high (n = 68) patients treated using sorafenib exhibited a significantly shorter progression-free survival and overall survival than FGF19low (n = 105) patients, there were no significant differences between FGF19high (n = 21) and FGF19low (n = 19) patients treated using lenvatinib. In conclusion, robust inhibition of FGF19/FGFR4 is of importance for the exertion of antitumor effects of MKIs. Serum FGF19 levels may function as a predictive marker for drug response and survival in HCC patients treated using sorafenib.

In Vitro Toxicology Methods in Risk Assessment

1996 ◽  
Vol 24 (3) ◽  
pp. 325-331
Author(s):  
Iain F. H. Purchase
Keyword(s):  
Risk Assessment ◽  
Hazard Assessment ◽  
Human Health Risk ◽  
In Vitro Test ◽  
In Vitro Methods ◽  
New Concepts ◽  
Vitro Test

The title of this paper is challenging, because the question of how in vitro methods and results contribute to human health risk assessment is rarely considered. The process of risk assessment usually begins with hazard assessment, which provides a description of the inherent toxicological properties of the chemical. The next step is to assess the relevance of this to humans, i.e. the human hazard assessment. Finally, information on exposure is examined, and risk can then be assessed. In vitro methods have a limited, but important, role to play in risk assessment. The results can be used for classification and labelling; these are methods of controlling exposure, analogous to risk assessment, but without considering exposure. The Ames Salmonella test is the only in vitro method which is incorporated into regulations and used widely. Data from this test can, at best, lead to classification of a chemical with regard to genotoxicity, but cannot be used for classification and labelling on their own. Several in vitro test systems which assess the topical irritancy and corrosivity of chemicals have been reasonably well validated, and the results from these tests can be used for classification. The future development of in vitro methods is likely to be slow, as it depends on the development of new concepts and ideas. The in vivo methods which currently have reasonably developed in vitro alternatives will be the easiest to replace. The remaining in vivo methods, which provide toxicological information from repeated chronic dosing, with varied endpoints and by mechanisms which are not understood, will be more difficult to replace.

scholarly journals Transcriptional Profiling of Porcine Blastocysts Produced In Vitro in a Chemically Defined Culture Medium

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1414
Author(s):  
Josep M. Cambra ◽  
Emilio A. Martinez ◽  
Heriberto Rodriguez-Martinez ◽  
Maria A. Gil ◽  
Cristina Cuello
Keyword(s):  
Culture Medium ◽  
Embryo Quality ◽  
Transcriptional Profiling ◽  
Defined Medium ◽  
Platelet Factor ◽  
Defined Media ◽  
Defined Culture ◽  
The Impact

The development of chemically defined media is a growing trend in in vitro embryo production (IVP). Recently, traditional undefined culture medium with bovine serum albumin (BSA) has been successfully replaced by a chemically defined medium using substances with embryotrophic properties such as platelet factor 4 (PF4). Although the use of this medium sustains IVP, the impact of defined media on the embryonic transcriptome has not been fully elucidated. This study analyzed the transcriptome of porcine IVP blastocysts, cultured in defined (PF4 group) and undefined media (BSA group) by microarrays. In vivo-derived blastocysts (IVV group) were used as a standard of maximum embryo quality. The results showed no differentially expressed genes (DEG) between the PF4 and BSA groups. However, a total of 2780 and 2577 DEGs were detected when comparing the PF4 or the BSA group with the IVV group, respectively. Most of these genes were common in both in vitro groups (2132) and present in some enriched pathways, such as cell cycle, lysosome and/or metabolic pathways. These results show that IVP conditions strongly affect embryo transcriptome and that the defined culture medium with PF4 is a guaranteed replacement for traditional culture with BSA.

scholarly journals Impact of RARα and miR-138 on retinoblastoma etoposide resistance

Tumor Biology ◽  
2021 ◽  
Vol 43 (1) ◽  
pp. 11-26
Author(s):  
Maike Busch ◽  
Natalia Miroschnikov ◽  
Jaroslaw Thomas Dankert ◽  
Marc Wiesehöfer ◽  
Klaus Metz ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Lines ◽  
Cancer Progression ◽  
Treated Patient ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Gene Assay ◽  
The Impact

BACKGROUND: Retinoblastoma (RB) is the most common childhood eye cancer. Chemotherapeutic drugs such as etoposide used in RB treatment often cause massive side effects and acquired drug resistances. Dysregulated genes and miRNAs have a large impact on cancer progression and development of chemotherapy resistances. OBJECTIVE: This study was designed to investigate the involvement of retinoic acid receptor alpha (RARα) in RB progression and chemoresistance as well as the impact of miR-138, a potential RARα regulating miRNA. METHODS: RARα and miR-138 expression in etoposide resistant RB cell lines and chemotherapy treated patient tumors compared to non-treated tumors was revealed by Real-Time PCR. Overexpression approaches were performed to analyze the effects of RARα on RB cell viability, apoptosis, proliferation and tumorigenesis. Besides, we addressed the effect of miR-138 overexpression on RB cell chemotherapy resistance. RESULTS: A binding between miR-138 and RARα was shown by dual luciferase reporter gene assay. The study presented revealed that RARα is downregulated in etoposide resistant RB cells, while miR-138 is endogenously upregulated. Opposing RARα and miR-138 expression levels were detectable in chemotherapy pre-treated compared to non-treated RB tumor specimen. Overexpression of RARα increases apoptosis levels and reduces tumor cell growth of aggressive etoposide resistant RB cells in vitro and in vivo. Overexpression of miR-138 in chemo-sensitive RB cell lines partly enhances cell viability after etoposide treatment. CONCLUSIONS: Our findings show that RARα acts as a tumor suppressor in retinoblastoma and is downregulated upon etoposide resistance in RB cells. Thus, RARα may contribute to the development and progression of RB chemo-resistance.

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