TY - JOUR AB - In the field of non-viral drug delivery, polyplexes (PXs) represent an advanced investigated and highly promising tool for the delivery of nucleic acids. Upon encountering physiological fluids, they adsorb biological molecules to form a protein corona (PC), that influence PXs biodistribution, transfection efficiencies and targeting abilities. In an effort to understand protein - PX interactions and the effect of PX material on corona composition, we utilized cationic branched 10 kDa polyethyleneimine (b-PEI) and a hydrophobically modified nylon-3 polymer (NM0.2/CP0.8) within this study to develop appropriate methods for PC investigations. A centrifugation procedure for isolating hard corona - PX complexes (PCPXs) from soft corona proteins after incubating the PXs in fetal bovine serum (FBS) for PC formation was successfully optimized and the identification of proteins by a liquid chromatography-tandem mass spectrometry (LC-MS-MS) method clearly demonstrated that the PC composition is affected by the underlying PXs material. With regard to especially interesting functional proteins, which might be able to induce active targeting effects, several candidates could be detected on b-PEI and NM0.2/CP0.8 PXs. These results are of high interest to better understand how the design of PXs impacts the PC composition and subsequently PCPXs-cell interactions to enable precise adjustment of PXs for targeted drug delivery. AU - Hartl, N.* AU - Jürgens, D.C.* AU - Carneiro, S.* AU - König, A.-C. AU - Xiao, X.* AU - Liu, R.* AU - Hauck, S.M. AU - Merkel, O.M.* C1 - 68075 C2 - 54553 CY - Radarweg 29, 1043 Nx Amsterdam, Netherlands TI - Protein corona investigations of polyplexes with varying hydrophobicity - From method development to in vitro studies. JO - Int. J. Pharm. VL - 643 PB - Elsevier PY - 2023 SN - 0378-5173 ER - TY - JOUR AB - Glucagon-like peptide-1 (GLP-1) has been considered to be a promising peptide for treatment of type 2 diabetes mellitus (T2DM). However, the extremely short half-life (minutes) of native GLP-1 limits its clinical application potential. Here, we designed two GLP-1 analogues by genetic fusion of GLP-1 to one or two tandem human serum albumin-binding designed ankyrin repeat proteins (DARPins), denoted as GLP-DARPin or GLP-2DARPin. The two DARPin-fusion GLP-1 proteins were expressed in E. coli and purified, followed by measurements of their bioactivities and half-lives in mice. The results revealed that the half-life of GLP-2DARPin, binding two HSA molecules, was approximately 3-fold longer than GLP-DARPin (52.3 h versus 18.0 h). In contrast, the bioactivity results demonstrated that the blood glucose-lowering effect of GLP-DARPin was more potent than that of GLP-2DARPin. The oral glucose tolerance tests indicated that blood glucose levels were significantly reduced for at least 48 hours by GLP-DARPin, but were reduced for only 24 h by GLP-2DARPin. Injected once every two days, GLP-DARPin substantially reduced blood glucose levels in streptozotocin (STZ)-induced diabetic mice to the same levels as normal mice. During the treatment course, GLP-DARPin significantly reduced the food intake and body weight of diabetic mice up to approximately 17% compared with the control group. A histological analysis revealed that GLP-DARPin alleviated islet loss in diabetic mice. These findings suggest that long-acting GLP-DARPin holds great potential for further development into drugs for the treatment of T2DM and obesity. Meanwhile, our data indicate that albumin-binding DARPins can be used as a universal scaffold to improve the pharmacokinetic profiles and pharmacological activities of therapeutic peptides and proteins. AU - Tan, H.* AU - Su, W.* AU - Zhang, W.* AU - Zhang, J.* AU - Sattler, M. AU - Zou, P. C1 - 62863 C2 - 51116 CY - Radarweg 29, 1043 Nx Amsterdam, Netherlands TI - Generation of novel long-acting GLP-1R agonists using DARPins as a scaffold. JO - Int. J. Pharm. VL - 607 PB - Elsevier PY - 2021 SN - 0378-5173 ER - TY - JOUR AB - Background and Objective: Diabetes Mellitus (DM) is a major healthcare problem worldwide and considerable evidence proved its negative impact on the male reproductive system. Adenium obesum is an interesting medicinal plant with a wide range of bioactivities. The current study examined the protective effects of A. obesum flower extract (AOE) on testicular injury in streptozotocin (STZ)-induced type I diabetic rats. Materials and Methods: Diabetes was induced by a single injection of 50 mg kg(-1) STZ. Diabetic rats received 250 and 500 mg kg(-1) AOE for 21 days and samples were collected for analysis. Results: As compared to the diabetic control rats, treatment with AOE increased serum testosterone, Follicle-Stimulating Hormone (FSH) and Luteinizing Hormone (LH) levels, decreased testicular thiobarbituric acid reactive substances (TBARS) content, effectively enhanced reduced glutathione (GSH) content and superoxide dismutase (SOD) activity. Additionally, AGE effectively inhibited diabetes-induced testicular tissue injuryand prevented inflammatoryand apoptotic responses manifested by decreased TNF-alpha, IL-6 and Bax and increased Bcl-2. Conclusion: These results demonstrated that AGE mitigates testicular injury, oxidative stress, inflammatory response and apoptotic cell death in STZ-induced diabetic rats. AU - Elgebaly, H.* AU - Germoush, M.* AU - Mosa, N.* AU - Zahou, F.* AU - Soffar, A.* AU - Alotaibi, N.* AU - Qarmush, M.* AU - Hussein, O.* AU - Bin-Jumah, M.* AU - Hassanein, E.* AU - Hernandez-Bautista, R. AU - Mahmoud, A.M.* C1 - 59287 C2 - 48716 CY - 308-lasani Town, Sargodha Rd, Faisalabad, 38090, Pakistan SP - 310-318 TI - Adenium obesum flowers extract mitigates testicular injury and oxidative stress in streptozotocin-induced diabetic rats. JO - Int. J. Pharm. VL - 16 IS - 4 PB - Asian Network Scientific Information-ansinet PY - 2020 SN - 0378-5173 ER - TY - JOUR AB - Aerosolized administration of biopharmaceuticals to the airways is a promising route for nasal and pulmonary drug delivery, but - in contrast to small molecules - little is known about the effects of aerosolization on safety and efficacy of biopharmaceuticals. Proteins are sensitive against aerosolization-associated shear stress. Tailored formulations can shield proteins and enhance permeation, but formulation development requires extensive screening approaches. Thus, the aim of this study was to develop a cell-based in vitro technology platform that includes screening of protein quality after aerosolization and transepithelial permeation. For efficient screening, a previously published aerosolization-surrogate assay was used in a design of experiments approach to screen suitable formulations for an IgG and an antigen-binding fragment (Fab) as exemplary biopharmaceuticals. Efficient, dose-controlled aerosol-cell delivery was performed with the ALICE-CLOUD system containing RPMI 2650 epithelial cells at the air-liquid interface. We could demonstrate that our technology platform allows for rapid and efficient screening of formulations consisting of different excipients (here: arginine, cyclodextrin, polysorbate, sorbitol, and trehalose) to minimize aerosolization-induced protein aggregation and maximize permeation through an in vitro epithelial cell barrier. Formulations reduced aggregation of native Fab and IgG relative to vehicle up to 50% and enhanced transepithelial permeation rate up to 2.8-fold. AU - Röhm, M.* AU - Carle, S.* AU - Maigler, F.* AU - Flamm, J.* AU - Kramer, V.* AU - Mavoungou, C.* AU - Schmid, O. AU - Schindowski, K.* C1 - 51905 C2 - 43553 CY - Amsterdam SP - 537-546 TI - A comprehensive screening platform for aerosolizable protein formulations for intranasal and pulmonary drug delivery. JO - Int. J. Pharm. VL - 532 IS - 1 PB - Elsevier Science Bv PY - 2017 SN - 0378-5173 ER - TY - JOUR AB - Succesful gene therapy requires stability and sufficient bioavailability of the applied drug at the site of action. In the case of RNA interference (RNAi), non-viral vectors play a promising role for delivering intact siRNA molecules. We selected a low molecular weight polyethyleneimine (PEI F25-LMW) and investigated the biokinetics of PEI F25-LMW/siRNA polyplexes in comparison to non-complexed siRNA molecules upon intratracheal application into mice. Additionally, a bronchoalveolar lavage was performed to locate the siRNA within the different lung compartments and to analyse possible inflammatory reactions. Liquid scintillation counting of a 32P-label was used to follow the siRNA within the whole body. During the complete observation time more than 75% of the applied dose was found at the target site. The complexation with PEI F25- LMW prevented the siRNA from being degraded and cleared and prolonged its retention time. A low inflammatory reaction was observed on the basis of cell differentiation. Taken together, PEI F25-LMW meets fundamental requirements on non-viral vectors for local pulmonary siRNA delivery. AU - Lipka, J. AU - Semmler-Behnke, M. AU - Wenk, A. AU - Burkhardt, J.* AU - Aigner, A.* AU - Kreyling, W.G. C1 - 47738 C2 - 39488 CY - Amsterdam SP - 227-235 TI - Biokinetic studies of non-complexed siRNA versus nano-sized PEI F25-LMW/siRNA polyplexes following intratracheal instillation into mice. JO - Int. J. Pharm. VL - 500 IS - 1-2 PB - Elsevier Science Bv PY - 2016 SN - 0378-5173 ER - TY - JOUR AB - Indocyanine green (ICG) is an FDA-approved, strongly photo-absorbent/fluorescent probe that has been incorporated into a clinically-relevant PEGylated liposome as a flexible optoacoustic contrast agent platform. This study describes the engineering of targeted PEGylated liposome-ICG using the anti-MUC-1 "humanized" monoclonal antibody (MoAb) hCTM01 as a tumour-specific theranostic system. We aimed to visualise non-invasively the tumour accumulation of these MoAb-targeted liposomes over time in tumour-bearing mice using multispectral optoacoustic tomography (MSOT). Preferential accumulation of targeted PEGylated liposome-ICG was studied after intravenous administration in comparison to non-targeted PEGylated liposome-ICG using both fast growing (4T1) and slow growing (HT-29) MUC-1 positive tumour models. Monitoring liposomal ICG in the tumour showed that both targeted and non-targeted liposome-ICG formulations preferentially accumulated into the tumour models studied. Rapid accumulation was observed for targeted liposomes at early time points mainly in the periphery of the tumour volume suggesting binding to available MUC-1 receptors. In contrast, non-targeted PEGylated liposomes showed accumulation at the centre of the tumour at later time points. In an attempt to take this a step further, we successfully encapsulated the anticancer drug, doxorubicin (DOX) into both targeted and non-targeted PEGylated liposome-ICG. The engineering of DOX-loaded targeted ICG liposome systems present a novel platform for combined tumour-specific therapy and diagnosis. This can open new possibilities in the design of advanced image-guided cancer therapeutics. AU - Lozano, N.* AU - Al-Ahmady, Z.S.* AU - Bézière, N. AU - Ntziachristos, V. AU - Kostarelos, K.* C1 - 42897 C2 - 35730 CY - Amsterdam SP - 2-10 TI - Monoclonal antibody-targeted PEGylated liposome-ICG encapsulating doxorubicin as a potential theranostic agent. JO - Int. J. Pharm. VL - 482 IS - 1-2 PB - Elsevier Science Bv PY - 2015 SN - 0378-5173 ER -