Julia Rohrer

Development and in vitro characterisation of an oral self-emulsifying delivery system for daptomycin

It was the aim of this study to develop an oral self-emulsifying drug delivery system (SEDDS) for the peptide drug daptomycin exhibiting an anionic net charge. Drug lipophilicity was increased by hydrophobic ion pairing with cationic surfactant dodecylamine hydrochloride in molar ratio of surfactant to peptide 5:1. Log P (octanol/water) of -5.0 was even raised to +4.8 due to complexation with dodecylamine hydrochloride. Various SEDDS formulations were developed and characterised regarding emulsification properties, droplet size, polydispersity index and zeta potential. When the daptomycin dodecylamine complex (DAP/DOA) was dissolved in a formulation containing 35% Dermofeel MCT, 30% Capmul MCM and 35% Cremophor RH40, a maximum payload of even 8.0% (w/w) corresponding to 5.5% pure daptomycin was achieved. The formulation was degraded by lipase within 90min. Release studies of daptomycin from this formulation emulsified in 50mM phosphate buffer pH6.8 demonstrated a sustained drug release for at least six hours. Moreover, SEDDS exhibited also mucus permeating properties as well as a protective effect towards drug degradation by α-chymotrypsin. According to these results, SEDDS containing 8% DAP/DOA complex may be considered as a new potential oral delivery system for daptomycin.

Impact of lipases on the protective effect of SEDDS for incorporated peptide drugs towards intestinal peptidases

AIM The aim of this study is the development of self-emulsifying drug delivery systems (SEDDS) differing in amounts of ester substructures and to evaluate their stability in presence of pancreatic lipase and protective effect against luminal enzymatic metabolism using leuprorelin as model peptide drug. METHODS Hydrophobic leuprolide oleate was incorporated into three different SEDDS formulations and their stability towards pancreatic lipases was investigated utilizing a dynamic in vitro digestion model. Protective effect of SEDDS in respect to peptide drug stability against proteolytic enzymes, trypsin and α-chymotrypsin, was determined via HPLC. RESULTS Results of in vitro digestion demonstrated that 80% of SEDDS containing the highest amount of ester linkages was degraded within 60min. In comparison to that, SEDDS without ester bonds showed no degradation. With increasing oil droplets hydrolysis the remaining amount of peptide encapsulated into formulation decreased. Furthermore, after 180min incubation with trypsin up to 33.5% and with α-chymotrypsin up to 60.5% of leuprolide oleate was intact while leuprorelin acetate aqueous solution was completely metabolized by trypsin within 120min and by α-chymotrypsin within 5min. Protective effect in environment containing lipases was lower due to oil phase degradation, however, the amount of peptide in ester-free SEDDS was remarkably higher compared to SEDDS susceptible to lipases. CONCLUSION The present study revealed that SEDDS stable towards hydrolysis is able to exhibit a protective effect for oral peptide delivery.

development, in vitro and in vivo evaluation of a self-emulsifying drug delivery system (sedds) for oral enoxaparin administration

AIM The aim of this study was to develop SEDDS for oral enoxaparin administration and evaluate it in vitro and in vivo. METHODS The emulsifying properties of SEDDS composed of long chain lipids (LC-SEDDS), medium chain lipids (MC-SEDDS), short chain lipids (SC-SEDDS) and no lipids (NL-SEDDS) were evaluated. Thereafter, enoxaparin was incorporated via hydrophobic ion pairing in the chosen SEDDS, which were evaluated regarding their mucus permeating properties, stability towards pancreatic lipase, drug release profile and cytotoxicity. Finally, in vivo performance of SEDDS was evaluated. RESULTS The average droplet size of chosen LC-SEDDS, MC-SEDDS and NL-SEDDS ranged between 30 and 40nm. MC-SEEDS containing 30% Captex 8000, 30% Capmul MCM, 30% Cremophor EL and 10% propylene glycol and NL-SEDDS containing 31.5% Labrafil 1944, 22.5% Capmul PG-8, 9% propylene glycol, 27% Cremophor EL and 10% DMSO exhibited 2-fold higher mucus diffusion than LC-SEDDS and were therefore chosen for further studies. The enoxaparin-dodecylamine complex (ENOX/DOA) was incorporated in a payload of 2% (w/w) into MC-SEDDS and NL-SEDDS. After 90min 97% of MC-SEDDS and 5% of NL-SEDDS were degraded by pancreatic lipase. Both MC-SEDDS and NL-SEDDS showed sustained in vitro enoxaparin release. Furthermore, orally administrated MC-SEDDS and NL-SEDDS yielded an absolute enoxaparin bioavailability of 2.02% and 2.25%, respectively. CONCLUSION According to the abovementioned findings, SEDDS could be considered as a potential oral LMWH delivery system.

Thiolated silicone oil: Synthesis, gelling and mucoadhesive properties

Graphical abstract

Preactivated thiolated poly(methacrylic acid-co-ethyl acrylate): Synthesis and evaluation of mucoadhesive potential

The study was aimed to developed and investigate a novel polymer for intestinal drug delivery with improved mucoadhesive properties. Therefore Eudragit® L 100-55 (poly(methacrylic acid-co-ethyl acrylate)) was thiolated by covalent attachment of L-cysteine. The immobilized thiol groups were preactivated by disulfide bond formation with 2-mercaptonicotinic acid. Resulting derivative (Eu-S-MNA) was investigated in terms of mucoadhesion via three different methods: tensile studies, rotating cylinder studies and rheological synergism method, as well as water-uptake capacity and cytotoxicity. Different derivatives were obtained with increasing amount of bound L-cysteine (60, 140 and 266 μmol/g polymer) and degree of preactivation (33, 45 and 51 μmol/g polymer). Tensile studies revealed a 30.5-, 35.3- and 52.2-fold rise of total work of adhesion for the preactivated polymers compared to the unmodified Eudragit. The adhesion time on the rotating cylinder was prolonged up to 17-fold in case of thiolated polymer and up to 34-fold prolonged in case of the preactivated polymer. Rheological synergism revealed remarkable interaction of all investigated modified derivatives with mucus. Further, water-uptake studies showed an over 7h continuing weight gain for the modified polymers whereat disintegration took place for the unmodified polymer within the first hour. Cell viability studies revealed no impact of modification. Accordingly, the novel preactivated thiolated Eudragit-derivative seems to be a promising excipient for intestinal drug delivery.

Mucus permeating thiolated self-emulsifying drug delivery systems.

CONTEXT Mucus represents a critical obstacle for self-emulsifying drug delivery systems (SEDDS) targeting the epithelial membrane site. OBJECTIVE The aim of the study was the development of a novel SEDDS to overcome the mucus barrier. MATERIALS AND METHODS Two novel conjugates N-dodecyl-4-mercaptobutanimidamide (thiobutylamidine-dodecylamine, TBA-D) and 2-mercapto-N-octylacetamide (thioglycolicacid-octylamine, TGA-O) were synthesized, incorporated into SEDDS and analyzed for stability, cytotoxicity and physico-chemical characteristics using dynamic light scattering. Mucus interaction studies were performed using in vitro assays based on multiple particle tracking, rotational silicone tubes and rheology. RESULTS AND DISCUSSION TBA-D was synthesized using dodecylamine and iminothiolane as thiol precursor (yield=55 ± 5%). TGA-O was obtained via crosslinking of octylamine with SATA ((2,5-dioxopyrrolidin-1-yl) 2-acetylsulfanylacetate) (yield=70 ± 6%). The chemical structure of target compounds was confirmed via NMR analysis. The thiol-conjugates were incorporated in an amount of 3% (m/m) into SEDDS (Cremophor EL 30%, Capmul MCM 30%, Captex 355 30% and propylene glycol 10%), namely thiolated SEDDS leading to a droplet size around 50 nm and zeta potential close to 0 mV. Thiolated SEDDS with an effective diffusion coefficient 〈Deff〉 of up to 0.871 ± 0.122 cm(2) s(-1) × 10(-9) were obtained. Rotational silicone studies show increased permeation of the thiolated SEDDS A in comparison with unthiolated control. Rheological studies confirmed the mucolytic activity of the thiol-conjugates which differed only by 3% from DTT (dithiothreitol) serving as positive control. CONCLUSION Low molecular weight thiol-conjugates were identified to improve the mucus permeation, leading to highly efficient mucus permeating SEDDS, which were superior to conventional SEDDS and might thus be a new carrier for lipophilic drug delivery.

Self-emulsifying drug delivery systems (SEDDS): Proof-of-concept how to make them mucoadhesive

Aim: The objective of this study was to provide a proof‐of‐concept that self‐emulsifying drug delivery systems can be made mucoadhesive by the incorporation of hydrophobic mucoadhesive polymers. Methods: In order to obtain such a hydrophobic mucoadhesive polymer, Eudragit® S100 was thiolated by covalent attachment of cysteamine. After determination of the thiol group content, in vitro mucoadhesion studies (rotating cylinder and rheological measurements) were performed. Then, synthesized conjugate was incorporated into self‐emulsifying drug delivery systems (SEDDS) and their toxic potential as well as that of unmodified and thiolated Eudragit® S100 was examined on Caco‐2 cell line. Lastly, the mucoadhesiveness of developed SEDDS on porcine intestinal mucosa was determined. Results: Generated thiolated Eudragit® S100 displaying 235 ± 14 &mgr;mol of free thiol groups and 878 ± 101 &mgr;mol of disulfide bonds per gram polymer showed a great improvement in both: dynamic viscosity with mucus and adhesion time on mucosal tissue compared to the unmodified polymer. Resazurin assay revealed that unmodified and thiolated polymers and also SEDDS dispersions were non‐toxic over Caco‐2 cells. Furthermore, the incorporation of 1.5% (w/w) of such thiomer into SEDDS led to remarkably improved mucoadhesiveness. Blank SEDDS were completely removed from the mucosa within 15 min, whereas >60% of SEDDS containing thiolated Eudragit® S100 were still attached to it. Conclusion: These results provide evidence that SEDDS can be made mucoadhesive by the incorporation of hydrophobic mucoadhesive polymers. Graphical abstract Figure. No caption available.

Thiolated silicone oils as adhesive skin protectants for improved barrier function.

The purpose of this study was the evaluation of thiolated silicone oil as novel skin protectant exhibiting prolonged residence time, enhanced barrier function and reinforced occlusivity.

Mucoadhesive polymers: Synthesis and in vitro characterization of thiolated poly(vinyl alcohol)

The aim of this study was to synthesize thiolated poly(vinyl alcohol) (PVA) and to evaluate its mucoadhesive properties. Thiourea and 3-mercaptopropionic acid were utilized in order to obtain thiolated PVAs, namely, TPVA1 and TPVA2, respectively. TPVA1 and TPVA2 displayed 130.44 ± 14.99 and 958.35 ± 155.27 μmol immobilized thiol groups per gram polymer, respectively, which were then evaluated regarding reactivity of thiol groups, swelling behavior and mucoadhesive properties. Both thiolated PVAs exhibited the highest reactivity at pH 8.0 whereas more than 95% of free thiol groups were preserved at pH 5.0. Thiolation of PVA decelerated water uptake and prolonged disintegration time of test discs compared to unmodified PVA. Contact time of TPVA1- and TPVA2-based test discs on porcine intestinal mucosa was 3.2- and 15.8-fold prolonged, respectively, in comparison to non-thiolated PVA as measured by rotating cylinder method. According to tensile studies on mucosa, the total work of adhesion (TWA) and the maximum detachment force (MDF) were increased when compared to PVA. Furthermore, thiolated PVAs preserved higher percentage of viable cells compared to unmodified PVA within 24h as evaluated by MTT assay. Accordingly, thiolated PVA represents a novel excipient that can likely improve the mucoadhesive properties of various pharmaceutical formulations.

Role of sonication pre-treatment and cation valence in the sol-gel transition of nano-cellulose suspensions

Sol-gel transition of carboxylated cellulose nanocrystals has been investigated using rheology, SAXS, NMR and optical spectroscopies to unveil the distinctive roles of ultrasound treatments and addition of various cations. Besides cellulose fiber fragmentation, sonication treatment induces fast gelling of the solution. The gelation is independent of the addition of cations, while the final rheological properties are highly influenced by the type, concentration and sequence of the operations since the cations must be added prior to sonication to produce stiff gels. The gel elastic modulus was found to increase proportionally to the ionic charge rather than the cationic size. In cases where ions were added after sonication, SAXS analysis of the Na+ hydrogel and Ca2+ hydrogel indicated the presence of structurally ordered domains in which water is confined, and 1H-NMR investigation showed the dynamics of water exchange within the hydrogels. Conversely, separated phases containing essentially free water were characteristic of the hydrogels obtained by sonication after Ca2+ addition, confirming that this ion induces irreversible fiber aggregation. The rheological properties of the hydrogels depend on the duration of the ultrasound treatments, enabling the design of programmed materials with tailored energy dissipation response.Sol-gel transition of carboxylated cellulose nanocrystals is investigated using rheology, SAXS, NMR and optical spectroscopies to unveil the distinctive roles of ultrasounds treatment and ions addition. Besides cellulose fibers fragmentation, sonication treatment induces fast gelling of the solution. Gelation is induced independently on the addition of cations, while the final rheological properties are highly influenced by the type, the concentration as well as on the sequence of the operations since salts must be added before sonication to produce stiff gels. Cations with various charge and dimension have been associated to ultrasounds to induce gelation and the gel elastic modulus increase proportionally with the charge over the ion size ratio. SAXS analysis of the Na + hydrogel and Ca 2+ hydrogel to which the ion was added after sonication shows the presence of structurally ordered domains where water is confined as indicated by 1H-NMR investigation of the dynamic of water exchange in the hydrogels. Conversely, separated phases containing essentially free water, characterize the hydrogels obtained by sonication after Ca 2+ addition, confirming that this ion induces irreversible fiber aggregation. The rheological properties of the hydrogels depend on the duration of the ultrasound treatment and it enables the design of materials programmed with tailored energy dissipation response. INTRODUCTION Nanocellulose (NC) is a renewable and biocompatible material with interesting and versatile properties which allow its integration in a huge number of applications, as has been extensively reviewed [1,2]. The procedures to break natural cellulose and obtain nano-sized structures are usually based on the combination of chemical modification or enzymatic hydrolysis with mechanical refinement [3,4]. Fine changes of these procedures give rise to different nanostructure morphology: branched nanofibrils with amorphous regions (CNF) and rod-like rigid nanocrystals (CNC) [5]. TEMPO-mediated oxidation of cellulose followed by sonication provides well dispersed, negatively charged CNC [6]. Despite the large interest on NC and its applications, several basic aspects regulating NC properties and its interaction with the environment are still unclear. Concerning the structure investigation, the effort has been addressed mainly toward the understanding of the liquid crystalline self-assembly resulting in ordered helical structures with peculiar mechanical and optical properties [7-9]. The self-assembly of NC or NC-composites into soft hydrogels [10,11] has been characterized in terms of macroscopic parameters such as mesh size, charge density, gelation rate, mechanical performances, or stability [12-15]. In this context, rheology experiments have been performed and a gel-like behavior of NC suspensions with an elastic response even at a low concentration [16] has been reported. In general, the rheological behaviour of NC suspensions is strongly dependent on NC production: mechanical fibrillation without chemical modification produces suspensions with flocculated structure while NC which underwent chemical processes, produces suspensions with better colloidal stability [17]. The static and dynamic rheological behavior of