Penetration Kinetics Research Articles

Formulation and evaluation of novel vesicular nanoethosomal patches for transdermal delivery of zidovudine: In vitro, ex vivo, and in vivo pharmacokinetic investigations

Ethosomes are novel soft malleable nanovesicles composed mainly of phospholipids, ethanol (relatively high concentration), glycol, and water. They are used for enhanced drug delivery through the skin. These zidovudine (ZV) vesicles were formulated by cold technique using phospholipids, cholesterol, propylene glycol, and ethanol. The nanoethosomal dispersion was transformed to transethosomal patches using HPMC E15 and chitosan as polymers, a mixture of dichloromethane (DCM) and methanol (1:1 ratio) as solvent, triethyl citrate as plasticizer, and span 80 as permeation enhancer. The physical features of the prepared patches were found to be within acceptable limits. The in vitro study showed drug diffusion of 71.25 − 88.19% through the semipermeable egg membrane in 24 hours. The ex vivo study showed drug permeation of 78.35 − 91.45% through the goat ear membrane in 24 hours. The combination of HPMC and EP4 transdermal patch was determined to be the most effective formulation for transdermal drug administration with sustained drug release, outperforming all other formulations based on acceptable physicochemical parameters, in vitro and ex vivo evaluation studies. Compared with the other formulations, the EP4 formulation had superior penetration kinetics, as proven by its highest permeation co-efficient (2.56 mg/cm2/h), flux (0.494 mg/cm2/h), and enhancement ratio (2.43). Moreover, dermal irritation tests demonstrated that topical therapy did not cause any irritation, indicating its safety. The in vivo bioavailability studies revealed that the AUC showed good bioavailability, which was approximately 188.99 times greater than that of the marketed product. According to the in vitro–in vivo correlation, the ex vivo (in vitro) data can represent physiological conditions that are exact replicas of those found in vivo. Therefore, the ZV transdermal patches exhibit enhanced drug permeation and good bioavailability for better therapeutic outcomes.

MRI on ion exchange resins at different length scales

AbstractIon exchange resins were studied on different length scales by magnetic resonance imaging (MRI) with the focus on their interactions with nanoparticles (NP) and molecular clusters. On the length scale of resin beds (bed diameters <20 mm), the behavior of NP and of molecular clusters was shown to depend on the kind of ion exchange resin and nanoscale moiety. The kinetics of absorption and penetration into the resin beads was quantified on a smaller length scale of a stack of resin beads (sample with an outer diameter of 1.7 mm). Finally, using an MRI μ‐coil (3D spatial resolution ≥8 μm), adsorption of superparamagnetic NP on individual resin beads was observed via the magnetic field disturbance characteristic for magnetic dipoles. As a result, this allows the detection of NP (diameter ≤100 nm) by MRI on much larger length scales of several micrometers.

Challenges of forward osmosis desalination processes using hydrogels as draw agents

The present study aims to investigate the properties, identification, and comprehension of the obstacles encountered in the forward osmosis process when utilizing a hydrogel drawing agent (FO-HG). Furthermore, a comparison is made between the FO-HG system and conventional forward osmosis systems employing a salt solution drawing agent. The comparison and evaluation of the swelling process of hydrogel and the kinetics of water penetration are conducted in both un-constrained and constrained states. Furthermore, the investigation and analysis are carried out to determine the presence or absence of internal concentration polarization (ICP) and external concentration polarization (ECP) phenomena. These phenomena are studied in situations with and without mixing, as well as in different orientations of the membrane (FO-mode and PRO-mode). The impact of these phenomena on the water flux of the systems FO-HG and FO-NaCl is also examined. An evaluation is conducted to determine the influence of the amount and size of hydrogel used as a draw agent in the FO-HG system on the water flux. The results of the study reveal that smaller hydrogel particles in the FO-HG system exhibit a higher flux compared to larger particles. Additionally, it is observed that the water flux in PRO-mode is unexpectedly higher when salt water is used as feed solution. This phenomenon can be ascribed to a counter-osmotic effect, originating from the FO state. Despite the high water absorption capacity of hydrogel and its potential as an ideal drawing agent in the forward osmosis process, the results demonstrate that the flux of the FO-HG system is inferior to that of the FO-NaCl system. Finally, the focus is on resolving the low flux issue by suggesting a process involving multiple cycles throughout day and night. We investigate the influence of hydrogel particle size, membrane surface, hydrogel layer thickness, as well as swelling and deswelling time in one cycle. The swelling time displays a peak at an optimal absorption duration, while the deswelling time does not show a similar optimal point, highlighting the difference between swelling and deswelling phenomena. Therefore, the hydrogels' high absorption capacity alone is insufficient for achieving desalination success. The research findings emphasize the high importance of synthesis of a membrane with minimal resistance, enabling a high water flux and suitable selectivity.

Preclinical evaluation of targeted therapies for central nervous system metastases.

The central nervous system (CNS) represents a site of sanctuary for many metastatic tumors when systemic therapies that control the primary tumor cannot effectively penetrate intracranial lesions. Non-small cell lung cancers (NSCLCs) are the most likely of all neoplasms to metastasize to the brain, with up to 60% of patients developing CNS metastases during the disease process. Targeted therapies such as tyrosine kinase inhibitors (TKIs) have helped reduce lung cancer mortality but vary considerably in their capacity to control CNS metastases. The ability of these therapies to effectively target lesions in the CNS depends on several of their pharmacokinetic properties, including blood-brain barrier permeability, affinity for efflux transporters, and binding affinity for both plasma and brain tissue. Despite the existence of numerous preclinical models with which to characterize these properties, many targeted therapies have not been rigorously tested for CNS penetration during the discovery process, whereas some made it through preclinical testing despite poor brain penetration kinetics. Several TKIs have now been engineered with the characteristics of CNS-penetrant drugs, with clinical trials proving these efforts fruitful. This Review outlines the extent and variability of preclinical evidence for the efficacy of NSCLC-targeted therapies, which have been approved by the US Food and Drug Administration (FDA) or are in development, for treating CNS metastases, and how these data correlate with clinical outcomes.

Imaging-Based Drug Penetration Profiling in an Excised Sheep Cornea Model.

Formulations designed to address ocular conditions and diseases are predominantly administered topically. While in vitro test systems have been developed to assess corneal permeation under extended contact conditions, methods focusing on determining the penetration depth and kinetics of a substance within the cornea itself rather than through it, are scarce. This study introduces a method for time-dependent penetration depth analysis (10 and 60 min) by means of a semiquantitative imaging method in comparison with a quantitative corneal depth-cut technique, employing fluorescein sodium at concentrations of 0.2 and 0.4 mg/mL as a small molecule model substance and sheep cornea as a human surrogate. Excised tissues exhibited sustained viability in modified artificial aqueous humor and maintained thickness (746 ± 43 µm) and integrity (electrical resistance 488 ± 218 Ω∙cm2) under the experimental conditions. Both methods effectively demonstrated the expected concentration- and time-dependent depth of penetration of fluorescein sodium, displaying a significantly strong correlation. The traceability of the kinetic processes was validated with polysorbate 80, which acted as a penetration enhancer. Furthermore, the imaging-based method enabled detecting the retention of larger structures, such as hyaluronic acid and nanoemulsions from the commercial eyedrop formulation NEOVIS® TOTAL multi, inside the lacrimal layer.

(Invited) Seeing the Sound: An Ultrasound-Mediated Intravascular Light Source for Noninvasive Brain Modulation

Light is used in a wide range of methods in biology and medicine, such as fluorescence imaging, optogenetics, photoactivatable gene editing, photothermal and photodynamic therapies to treat cancers, and photochemotherapy to inactivate viruses in vivo. A critical challenge of applying light in vivo, such as deep-brain optogenetic neuromodulation and photochemotherapy in deep organs, arises from the poor penetration of photons in biological tissue due to scattering and absorption. Therefore, delivering light deep into the body requires invasive procedures, such as the insertion of optical fibers and endoscopes, as well as surgical removal of overlying tissues. The very invasiveness of these procedures also precludes easy repositioning and volume adjustment of the illuminated region in the same subject. To address these challenges, our lab has developed an ultrasound-mediated intravascular light source, leveraging the deep-tissue penetration of focused ultrasound. We capitalized on mechanoluminescent nanotransducers (MLNTs), which are colloidal nanoparticles of mechanoluminescent materials synthesized via a biomineral-inspired suppressed dissolution approach. These MLNTs can be delivered intravenously into blood circulation and emit light locally at the ultrasound focus. Owing to the deep penetration and fast temporal kinetics of ultrasound, we have demonstrated that this method can produce on-demand and dynamically programmable light emission patterns at elevated depths in different organs of live mice with millisecond precision. This ultrasound-mediated intravascular light source has allowed us to perform noninvasive “sono-optogenetic” neuromodulation in live mice, as well as brain-wide “scanning optogenetics” that activate different brain regions of the same mouse brain. Our development of the ultrasound-mediated intravascular light source has been published in PNAS (2019), Science (2020), Science Advances (2022), JACS (2022), JACS (2023), and Nature Protocols (2023).

(Invited) Non-Invasive Injectable Nanotube-Hydrogel Formulations for Local Analyte Detection

Rapid, minimally-invasive in situ detection of analytes in vivo remains a goal to study inflammatory signaling molecules, disease biomarkers, and pharmacology of therapeutics. To achieve this, we are using the near-infrared fluorescent properties of single-walled carbon nanotubes (SWCNT) to design sensors for use in live mice. SWCNT fluorescence is responsive to the local environment, in the tissue-transparent window, and does not photobleach, allowing for long-term signal monitoring. In this work, we are exploring several formulations of SWCNT embedded within hydrogels which gel in situ. We have investigated the kinetics of penetration of several classes of analytes—proteins, small molecules, and ions—through the hydrogel and their interaction with SWCNT sensors in vitro. Then, we explored the long-term stability of these nanotube-hydrogel formulations in live mice prior to detection of the chemotherapeutic doxorubicin. In ongoing work, we are exploring disease biomarker-specific sensing and monitoring of inflammatory cytokines in mouse models of disease using this nanotube-hydrogel system. We anticipate long-term applications of this work may include minimally-invasive and rapid clinical diagnostic development.

Topical calcineurin and mammalian target of rapamycin inhibitors in inflammatory dermatoses: Current challenges and nanotechnology‑based prospects (Review).

Topical therapy remains a critical component in the management of immune‑mediated inflammatory dermatoses such as psoriasis and atopic dermatitis. In this field, macrolactam immunomodulators, including calcineurin and mammalian target of rapamycin inhibitors, can offer steroid‑free therapeutic alternatives. Despite their potential for skin‑selective treatment compared with topical corticosteroids, the physicochemical properties of these compounds, such as high lipophilicity and large molecular size, do not meet the criteria for efficient penetration into the skin, especially with conventional topical vehicles. Thus, more sophisticated approaches are needed to address the pharmacokinetic limitations of traditional formulations. In this regard, interest has increasingly focused on nanoparticulate systems to optimize penetration kinetics and enhance the efficacy and safety of topical calcineurin and mTOR inhibitors in inflamed skin. Several types of nanovectors have been explored as topical carriers to deliver tacrolimus in both psoriatic and atopic skin, while preclinical data on nanocarrier‑based delivery of topical sirolimus in inflamed skin are also emerging. Given the promising preliminary outcomes and the complexities of drug delivery across inflamed skin, further research is required to translate these nanotherapeutics into clinical settings for inflammatory skin diseases. The present review outlined the dermatokinetic profiles of topical calcineurin and mTOR inhibitors, particularly tacrolimus, pimecrolimus and sirolimus, focusing on their penetration kinetics in psoriatic and atopic skin. It also summarizes the potential anti‑inflammatory benefits of topical sirolimus and explores novel preclinical studies investigating dermally applied nanovehicles to evaluate and optimize the skin delivery, efficacy and safety of these 'hard‑to‑formulate' macromolecules in the context of psoriasis and atopic dermatitis.

Effect of Exposure Conditions on Mortar Subjected to an External Sulfate Attack.

This study aims to investigate the influence of exposure conditions on the behavior of mortar subjected to an external sulfate attack (ESA). Three different exposure conditions (full immersion, semi-immersion, and drying/wetting cycles) were tested on mortar prisms made with Portland cement and two w/c ratios (0.45 and 0.6). To monitor degradation, it was necessary to evaluate variations in length (expansion), mass changes, compressive and tensile strengths, changes in the total porosity measured using water accessible porosity tests, and changes in the macroscopic behavior of the samples. Mercury intrusion porosimetry (MIP) was used to determine the size distribution of the pores. It was demonstrated that mixing mortar with the lower w/c ratio of 0.45 results in improved performance against an ESA. This study also demonstrates that the type of exposure to an ESA has no significant effect on the kinetics of sulfate penetration during the exposure period. However, the sample's surface becomes more cracked when subjected to repeated drying and wetting cycles. For all the considered exposure conditions, expansion occurred in three stages. In stage 1, the reaction product (ettringite) precipitated in large voids, without causing significant expansion (the expansion remained low and stable). During the second stage, the reaction products generated growing internal stress. The final stage of expansion resulted in microcracks, strength losses, and the formation of macropores, which ultimately lead to material failure. The MIP results indicate that major changes in the porosity and pore volume distribution occur at the surface layer in regard to the gel and capillary pore ranges.

An experimental study on the impact of particle surface wettability on melt infiltration in particulate beds

Melt infiltration into porous media is an intriguing phenomenon that holds immense significance across various sciences and technologies. In this work, the problem of metallic melt infiltration in particulate beds is investigated for understanding and prediction of severe accident progression associated with a molten pool penetrating through an underlying debris bed which may form in the reactor core or in the lower head of a light water reactor.The present study aims to quantify the effect of particle surface’s wettability on melt infiltration kinetics. For this purpose, two categories of experiment are conceived and carried out to measure the wettability of different material surfaces by melt and to characterize melt infiltration kinetics in one-dimensional particulate beds, respectively. The melt material is tin–bismuth eutectic alloy with a melting point of 139 °C. Copper (Cu), stainless steel (SS), Tin (Sn) and tin-coated stainless steel (Sn-coated SS) are chosen as materials of substrates and particles in wettability measurement and melt infiltration study. The particulate beds, packed with 1.5 mm spheres, are preheated to 200 °C before the melt infiltration begins.The experimental data of wettability measurement shows that the contact angles of liquid Sn-Bi eutectic on the above-mentioned material surfaces range from 79° to 135°. The results of melt infiltration tests confirm the significant effect of wettability on melt penetration kinetics. The capillary force plays a significant role in the initial infiltration of particulate beds. Specifically, a wettable particulate bed enhances the initial melt infiltration, whereas non-wettable beds hinder it.

Diffusion kinetics and perfusion times in tissue models obtained by bioorthogonal Raman μ-spectroscopy

The penetration kinetics of small-molecule compounds like nutrients, drugs, and cryoprotective agents into artificial cell aggregates are of pivotal relevance in many applications, from stem cell differentiation and drug screening through to cryopreservation. Depending on compound and tissue properties as well as aggregate size and shape, the penetration behavior can differ vastly. Here, we introduce bioorthogonal Raman microspectroscopy as a contactless technique to investigate the penetration of various compounds into spheroids, organoids, and other tissue models in terms of diffusion coefficients and perfusion times. We showcase the potential of the method by applying it to the radial perfusion of neural stem cell spheroids with the prevalent cryopreservation additive dimethyl sulfoxide. Employing a diffusion model for spherical bodies, the spectroscopic data were quantitatively analyzed. Perfusion times were obtained for spheroids in the sub-mm region, and interesting findings about the spheroid-size dependence of the diffusion coefficient are reported.

Infrared Spectroscopy Study of Edge Water Penetration at Hydrophilic Bonding Interface

Hydrophilic wafer bonding is a process used to bond two wafers without adding material between the two surfaces. Water has a definite impact both during the bonding (bonding wave propagation, adhesion) (1) and after annealing (mechanical strength of the bonding interface, adherence) (2) during such a process. To have a better understanding of mechanisms ruling direct hydrophilic wafer bonding, it is thus important to quantify the amount of water present at the bonding interface.It is known that water is able to penetrate at the bonding interface at room temperature and 40% relative humidity. This has been demonstrated by analyzing defects with scanning acoustic microscopy (SAM) and from X-Ray Reflectivity (XRR) measurements (3,4). Kinetics observed by Tedjini et al. were consistent with a Lucas-Washburn flow through a one nm gap driven by the capillary forces of the hydrophilic surfaces.Nevertheless, no investigation of this phenomenon was performed using Fourier Transform Infra-Red Multiple Internal Reflection (FTIR-MIR) spectroscopy (5). Infrared spectroscopy of silanols, hydrogen bonded water and free water is well documented (6–8) and provides knowledge about water content and organization at the bonding interface. Here FTIR-MIR spectroscopy yields a direct measurement of the relative amount of water and its structure at the interface.More precisely this study will enable us to compare Si-Si, Si-SiO2 and SiO2-SiO2 water penetration kinetics at a fixed distance from the edge of the bonded substrates. A schematic view of the FTIR-MIR setup is shown in Figure 1 to explain the measurement principle.We report the study of water kinetics penetration at bonding interface by FTIR-MIR characterization. A Gaussian decomposition of the OH stretching absorption band signal allows identifying the contributions of silanols, surface bonded water and free liquid water as shown on Figure 2. The integral of each Gaussian peak is otherwise proportional to the relative amount of water. Surface preparation will be discussed and results compared to other characterizations techniques.REFERENCES Larrey V, Mauguen G, Fournel F, Radisson D, Rieutord F, Morales C, et al. Adhesion Energy and Bonding Wave Velocity Measurements. ECS Transactions. 23 sept 2016;75(9):145-52.Fournel F, Tedjini M, Larrey V, Rieutord F, Morales C, Bridoux C, et al. Impact of Water Edge Absorption on Silicon Oxide Direct Bonding Energy. ECS Transactions. 23 sept 2016;75(9):129-34.Tedjini M, Fournel F, Moriceau H, Larrey V, Landru D, Kononchuk O, et al. Interface water diffusion in silicon direct bonding. Appl Phys Lett. 12 sept 2016;109(11):111603.Rieutord F, Tardif S, Landru D, Kononchuk O, Larrey V, Moriceau H, et al. Edge Water Penetration in Direct Bonding Interface. ECS Trans. 24 août 2016;75(9):163-7.Rochat N, Olivier M, Chabli A, Conne F, Lefeuvre G, Boll-Burdet C. Multiple internal reflection infrared spectroscopy using two-prism coupling geometry: A convenient way for quantitative study of organic contamination on silicon wafers. Appl Phys Lett. 2 oct 2000;77(14):2249-51.Baum M, Rébiscoul D, Juranyi F, Rieutord F. Structural and Dynamical Properties of Water Confined in Highly Ordered Mesoporous Silica in the Presence of Electrolytes. J Phys Chem C. 30 août 2018;122(34):19857-68.Caër SL, Pin S, Esnouf S, Raffy Q, Ph. Renault J, Brubach JB, et al. A trapped water network in nanoporous material: the role of interfaces. Physical Chemistry Chemical Physics. 2011;13(39):17658-66.Davis KM, Tomozawa M. An infrared spectroscopic study of water-related species in silica glasses. Journal of Non-Crystalline Solids. 2 juin 1996;201(3):177-98. Figure 1

Abstract B157: Preclinical evaluation of the blood-brain barrier permeability and intracranial efficacy of the next-generation RET inhibitor Vepafestinib

Abstract Background. Drug resistance and central nervous system (CNS) metastasis often hampers response to precision therapy for RET-rearranged solid tumors. Vepafestinib (Vepa) is a clinical-stage, potent and selective RET tyrosine-kinase inhibitor (TKI) with proven activity against on-target resistance mutations that arise post first-generation RET TKI. Here, we utilized an array of structural and pharmacokinetic methods to determine blood-brain barrier (BBB) permeability of vepa compared with other RET TKIs. Furthermore, we correlate BBB penetrability of RET inhibitors with in vivo efficacy in preclinical models of RET-rearranged intracranial solid tumors. Methods. RET inhibitors were generated by rational chemical design to identify agents which penetrate BBB more effectively than currently approved RET TKIs, and with reduced efflux transporter susceptibility. Transcellular transport assays were conducted in LLC-PK1 and MDCK II cells expressing P-gp and BCRP, respectively. Brain penetrability was assessed by total (Kp) and unbound (Kp,uu) brain:plasma concentration ratios in male Balb/c mice and microdialysis studies in freely-moving male Wistar rats. Models of CNS metastasis were generated by implanting cells labelled with luciferase into the cerebellum of mice. Studies were conducted in comparison with selpercatinib (Selp), pralsetinib (Pral) and TPX-0046. Results. Assessment of Kp and Kp,uu parameters of analogs revealed an excellent structure-activity relationship where modification at the 6-position of the pyrrolopyrimidine core resulted in improved BBB penetrability. The analog with the most favorable physiochemical and biological properties was Vepa. Vepa showed poor susceptibility to P-gp and BCRP-mediated efflux and better CNS penetrability (Kp,uu: 1.3) and retention than Sel, Pral and TPX-0046. The ratio of the observed concentrations of Vepa in microdialysates from the prefrontal cortex, cerebrospinal fluid (CSF) and plasma free fraction was approximately 1:1:1; these ratios were maintained from 2h to 6.5h after Vepaadministration (up to 8h for CSF). Importantly, Vepa was more effective than Selp at causing regression of intracranial xenograft tumors of RET fusion-driven malignancies (lung adenocarcinoma and sarcoma), thus extending survival of tumor-bearing animals. Common first-generation RET TKI resistance mutations at the solvent front (G810A/C/S/R), gatekeeper (V804L/M), hinge (Y806C) and roof (L730Q/R) regions remained sensitive to Vepa. Vepa was more selective than Selp, Pral and TPX-0046, inhibiting only RET when profiled against a panel of 256 kinases. In contrast, Selp, Pral and TPX-0045 also inhibited VEGFR2, and 4, 11 and 39 additional kinases, respectively. Conclusions. Vepafestinib is a structurally distinct and specific RET inhibitor, with superior brain penetration and retention kinetics. It is more effective than selpercatinib in controlling CNS disease. Vepafestinib is currently undergoing phase 1 and 2 clinical trial for patients with advanced solid tumors with RET alterations (margaRET, NCT04683250). Citation Format: Igor Odintsov, Kentaro Wakayama, Tom Zhang, Satoru Iguchi, Masanori Kato, Allan JW Lui, Inna Khodos, Morana Vojnic, Claudio Giuliano, Annalisa Bonifacio, Monika A Davare, Elisa de Stanchina, Emanuela Lovati, Marc Ladanyi, Isao Miyazaki, Romel Somwar. Preclinical evaluation of the blood-brain barrier permeability and intracranial efficacy of the next-generation RET inhibitor Vepafestinib [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr B157.

Boost of cosmetic active ingredient penetration triggered and controlled by the delivery of kHz plasma jet on human skin explants

This work reports on the demonstration of the penetration of cosmetic active ingredients (caffeine and hyaluronic acid) in human skin explants following safe and controlled plasma jet exposure. First, temperature increase and immunohistochemistry in the stratum corneum and epidermis were characterized to check the safe delivery of plasma jets and to select two operation regimes at 1 and 20 kHz. Plasma exposure for tens of seconds is shown to induce transient modulations of skin pH, transepidermal water loss, and skin wettability, revealing a reversible skin barrier function modulation. Then, it is demonstrated that plasma exposure significantly accelerates the penetration of active ingredients. The tuning of the plasma jet pulse repetition rate allows controlling the penetration kinetics. Such ex vivo results agree with previous in vitro experiments also exhibiting a transient permeabilization time window. A preliminary demonstration of human skin wettability modulation with a low-power, user-friendly dielectric barrier discharge setup is documented, opening perspectives for plasma-based home cosmetic care device development. To the best of our knowledge, this work is one of the first demonstrations of safe and controlled plasma-assisted active ingredients’ skin penetration in the context of cosmetic applications.

Evaluation of the 18F-labeled analog of the therapeutic all-D-enantiomeric peptide RD2 for amyloid β imaging

Positron emission tomography (PET) imaging with radiotracers that bind to fibrillary amyloid β (Aβ) deposits is an important tool for the diagnosis of Alzheimer's disease (AD) and for the recruitment of patients into clinical trials. However, it has been suggested that rather than the fibrillary Aβ deposits, it is smaller, soluble Aβ aggregates that exert a neurotoxic effect and trigger AD pathogenesis. The aim of the current study is to develop a PET probe that is capable of detecting small aggregates and soluble Aβ oligomers for improved diagnosis and therapy monitoring. An 18F-labeled radioligand was prepared based on the Aβ-binding d-enantiomeric peptide RD2, which is currently being evaluated in clinical trials as a therapeutic agent to dissolve Aβ oligomers. 18F-labeling was carried out using palladium-catalyzed S-arylation of RD2 with 2-[18F]fluoro-5-iodopyridine ([18F]FIPy). Specific binding of [18F]RD2-cFPy to brain material from transgenic AD (APP/PS1) mice and AD patients was demonstrated with in vitro autoradiography. In vivo uptake and biodistribution of [18F]RD2-cFPy were evaluated using PET analyses in wild-type and transgenic APP/PS1 mice. Although brain penetration and brain wash-out kinetics of the radioligand were low, this study provides proof of principle for a PET probe based on a d-enantiomeric peptide binding to soluble Aβ species.

Visualization of mechanical forces in 3D cell models with molecular DNA tension probe.

Mechanical force can act as an important stimulus to regulate cellular processes. Accurate measurement of these forces plays a fundamental role in our understanding of cell mechanosensing and mechanotransduction. Current methods have focused on investigating the cell-matrix and intercellular forces on 2D monolayer cell models. In contrast, cells in real biological systems exist in multilayers and expose to forces from 3D directions. Exploring cellular mechanical forces in 3D cell models is thus critical to reveal their native interaction patterns in organs or tissues. We have recently developed a type of lipid-DNA-based molecular tension probes that have emerged as a potent tool for imaging intercellular mechanical forces. In this work, we have further investigated the potential usage of these molecular DNA tension probes for measuring mechanical forces in 3D spheroids and organoids. Using tumor spheroids and stem cell organoids of different sizes and shapes, we first characterized and optimized the probe penetration and cell membrane modification kinetics and efficiencies of the lipid-DNA conjugates. Afterwards, as a proof of concept, E-cadherin-mediated intercellular tensile forces were imaged and quantified in these 3D cell models using designer ratiometric DNA tension probes. Such membrane tensile force measurement was enabled because these DNA hairpin-based probes can convert cellular mechanical forces into digital fluorescence signals. These optimized DNA tension probes can also be used to investigate the dynamic changes and spatial distributions of different cellular mechanotransduction processes. We believe this is an important study that can potentially fill the gap between current studies of 2D cell mechanics and that in real 3D biological models.

Assessment of penetration and permeation of caffeine by confocal Raman spectroscopy in vivo and ex vivo by tape stripping.

Tape stripping is an often-used non-invasive destructive method to investigate the skin penetration of a substance. In recent years, however, the suitability of confocal Raman spectroscopy (CRS) as a non-invasive method of non-destructive examination of the skin has become increasingly apparent. In this study, we compared invasion and depletion penetration and permeation kinetics of a 2% caffeine solution with and without 1,2-pentanediol as a penetration enhancer measured with CRS and tape stripping. Porcine skin was used for tape stripping and human skin for CRS. 2% caffeine solution was applied to the skin for different incubation times. Human skin was then examined by CRS while caffeine was extracted from porcine skin and quantified via reverse-phase HPLC. Fluxes were also measured and calculated by sum of the total amounts of caffeine penetrated into the skin. Without penetration enhancers, there is hardly any difference between the penetration profiles of the two measurement methods for invasion, but the curves for depletion are different. Furthermore, the calculated flux values for the invasion are almost identical, but for the depletion the tape stripping values are about twice as high as the CRS values. The relevance of conducting invasion and depletion studies became clear and was able to show the still existing problems in the comparability of CRS and tape stripping.

MA13.05 TA0953/HM06, a Novel RET-specific Inhibitor Effective in Extracranial and CNS Disease Models of NSCLC with RETfusions

RET kinase fusions act as oncogenic drivers in approximately 2% of NSCLC patients. Two RET-selective kinase inhibitors, selpercatinib and pralsetinib, have proven effective at reducing tumor burden, and are FDA-approved. However, the emergence of resistance to these drugs, and brain metastasis, remain serious clinical challenges. TAS0953/HM06 is a new pharmacologically advanced RET-specific inhibitor that is structurally different from other RET inhibitors, has superior brain penetration kinetics compared to selpercatinib and pralsetinib, and retains activity in the setting of RET G810 (solvent front) mutations.

Investigation of the filling of a porous ceramic matrix by molten salts using nano X-ray tomography

High-resolution nano X-ray computed tomography (nano-XCT) was used to investigate the process of infiltration of a Molten Carbonate Fuel Cell (MCFC) matrix by liquid electrolyte. A ceramic matrix made of a LiAl 2O3 powder was infiltrated by a liquid electrolyte during the start-up process of the MCFC. Nano-XCT extends the state-of-the art techniques for characterizing the morphology of the ceramic matrix and for the study of the penetration kinetics with electrolyte. Quantitative morphology analysis using high-resolution nano-XCT provides 3D information for each MCFC component nondestructively. The spatially resolved volume fraction of the molten electrolyte calculated from the 3D XCT data is used for simulation and to improve MCFC performance. Based on the results obtained, a new start-up procedure is proposed for a single MCFC. This procedure is consistent with the morphology changes which occur during the melting process inside the ceramic matrix.

Comparison Between Franz Diffusion Cell and a novel Micro-physiological System for In Vitro Penetration Assay Using Different Skin Models.

In vitro diffusive models are an important tool to screen the penetration ability of active ingredients in various formulations. A reliable assessment of skin penetration enhancing properties, mechanism of action of carrier systems, and an estimation of a bioavailability are essential for transdermal delivery. Given the importance of testing the penetration kinetics of different compounds across the skin barrier, several in vitro models have been developedThe aim of this study was to compare the Franz Diffusion Cell (FDC) with a novel fluid-dynamic platform (MIVO) by evaluating penetration ability of caffeine, a widely used reference substance, and LIP1, a testing molecule having the same molecular weight but a different lipophilicity in the two diffusion chamber systems. A 0.7% caffeine or LIP1 formulation in either water or propylene glycol (PG) containing oleic acid (OA) was topically applied on the Strat-M® membrane or pig ear skin, according to the infinite-dose experimental condition (780 ul/cm2). The profile of the penetration kinetics was determined by quantify the amount of molecule absorbed at different time-points (1, 2, 4, 6, 8 hours), by means of HPLC analysis. Both diffusive systems show a similar trend for caffeine and LIP1 penetration kinetics. The Strat-M® skin model shows a lower barrier function than the pig skin biopsies, whereby the PGOA vehicle exhibits a higher penetration, enhancing the effect for both diffusive chambers and skin surrogates. Most interestingly, MIVO diffusive system better predicts the lipophilic molecules (i.e. LIP1) permeation through highly physiological fluid flows resembled below the skin models.