A5010694970- Receptor Mechanisms and Signaling
- Chemical Synthesis and Analysis
- Neuropeptides and Animal Physiology
- Mast cells and histamine
- Computational Drug Discovery Methods
- Bioinformatics and Genomic Networks
- Neuroscience and Neuropharmacology Research
- Protein Structure and Dynamics
- Monoclonal and Polyclonal Antibodies Research
- Gene Regulatory Network Analysis
- Pharmacological Receptor Mechanisms and Effects
- Drug Transport and Resistance Mechanisms
- Circadian rhythm and melatonin
- DNA and Nucleic Acid Chemistry
- Pharmacological Effects and Assays
- vaccines and immunoinformatics approaches
- Coagulation, Bradykinin, Polyphosphates, and Angioedema
- Pancreatic function and diabetes
- Biochemical and Structural Characterization
- Cell Image Analysis Techniques
- PARP inhibition in cancer therapy
- Asthma and respiratory diseases
- Innovative Microfluidic and Catalytic Techniques Innovation
- Ion channel regulation and function
- Genetics, Bioinformatics, and Biomedical Research
University of Regensburg
2013-2021
Leibniz Supercomputing Centre
2021
Bavarian Academy of Sciences and Humanities
2021
The race to meet the challenges of global pandemic has served as a reminder that existing drug discovery process is expensive, inefficient and slow. There major bottleneck screening vast number potential small molecules shortlist lead compounds for antiviral development. New opportunities accelerate lie at interface between machine learning methods, in this case, developed linear accelerators, physics-based methods. two
Derivatization of biologically active peptides by conjugation with fluorophores or radionuclide-bearing moieties is an effective and commonly used approach to prepare molecular tools diagnostic agents. Whereas lysine, cysteine, N-terminal amino acids have been mostly for peptide conjugation, we describe a new, widely applicable based on the nonclassical bioisosteric replacement guanidine group in arginine functionalized carbamoylguanidine moiety. Four arginine-containing receptor ligands...
The drug discovery process currently employed in the pharmaceutical industry typically requires about 10 years and $2–3 billion to deliver one new drug. This is both too expensive slow, especially emergencies like COVID-19 pandemic. In silico methodologies need be improved select better lead compounds, so as improve efficiency of later stages protocol, identify those compounds more quickly. No known methodological approach can this combination higher quality speed. Here, we describe an...
In contrast to the corresponding mouse and rat orthologs, human histamine H4 receptor (hH4R) shows extraordinarily high constitutive activity. extracellular loop (ECL), replacement of F169 by V as in H4R significantly reduced Stabilization inactive state was even more pronounced for a double mutant, which, addition F169V, S179 ligand binding site replaced M. To study role FF motif ECL2, we generated hH4R-F168A mutant. The co-expressed Sf9 insect cells with G-protein subunits Gαi2 Gβ1γ2,...
Background and Purpose Some histamine H 4 receptor ligands act as inverse agonists at the human ( hH R ), a with exceptionally high constitutive activity, but neutral antagonists or partial constitutively inactive mouse mH ) rat rH ). To study molecular determinants of reciprocal mutants were constructed: single mutants: ‐ F 169 V , 171 S 179 A M ; double + 181 . Experimental Approach Site‐directed mutagenesis p VL 1392 plasmids containing receptors performed. Wild‐type mutant co‐expressed G...
On the basis of long-known prototypic pharmacophore 3-(1H-imidazol-4-yl)propylguanidine (SK&F 91486, 2), monomeric, homodimeric, and heterodimeric bisalkylguanidine-type histamine H2 receptor (H2R) agonists with various alkyl spacers were synthesized. Aiming at increased H2R selectivity ligands, imidazol-4-yl moiety was replaced by imidazol-1-yl, 2-aminothiazol-5-yl or 2-amino-4-methylthiazol-5-yl according to a bioisosteric approach. All compounds turned out be partial full h/gp/rH2R. The...
An index of the activation Class A G-protein-coupled receptors (GPCRs) has been trained using interhelix distances from a series microsecond molecular-dynamics simulations and tested for 268 published X-ray structures. In three-class model that includes intermediate structures, 63% active structures are classified in agreement with experimental assignment, 81% 89% inactives. alternative two-state classifies 94% actives 99% inactives correctly. The distributed 2:1 between protein nanobodies...
Comprehensively characterized fluorescent probes for the histamine H3 receptor (H3R) and especially H4R orthologs [e.g., human (h) mouse (m)] are highly needed as versatile complementary tools to radioligands. In view of BRET-based binding studies localizing in live cells, we synthesized biologically Py-5-labeled derivatives. The most notable compound was UR-DEBa242 (26, 1-[4-(1H-Imidazol-4-yl)butyl]-4-{(1E,3E)-4-[4-(dimethylamino)phenyl]buta-1,3-dienyl}-2,6-dimethylpyridinium...
The investigation of the (patho)physiological role histamine H4 receptor (H4R) and its validation as a possible drug target in translational animal models are compromised by distinct species-dependent discrepancies regarding potencies subtype selectivities pharmacological tools. Such differences were extremely pronounced case proximal readouts, e. g. [32P]GTPase or [35S]GTPγS binding assays. To improve predictability vitro investigations, aim this study was to establish reporter gene assay...
The dualsteric ligand approach, aiming at ligands with improved subtype selectivity, has been increasingly applied to muscarinic receptors (MRs). In this article, we present the synthesis and characterization of a M2R subtype-preferring radiolabeled dibenzodiazepinone-type antagonist ([3H]UNSW-MK259, [3H]19) its homodimeric analogue [3H]UR-AP060 ([3H]33). Saturation binding studies M2R, using orthosteric atropine determine unspecific binding, proved that monomeric dimeric compound bind site...
Fluorescently labeled dibenzodiazepinone-type muscarinic acetylcholine receptor (MR) antagonists, including dimeric ligands, were prepared using red-emitting cyanine dyes. Probes containing a fluorophore with negative charge showed high M2R affinities (pKi (radioligand competition binding): 9.10-9.59). Binding studies at M1 and M3-M5 receptors indicated preference. Flow cytometric high-content imaging saturation binding (M1R, M2R, M4R) confirmed occupation of the orthosteric site. Confocal...
The family of neuropeptide Y (NPY) receptors comprises four subtypes (Y1R, Y2R, Y4R, Y5R), which are addressed by at least three endogenous peptides, i.e., NPY, peptide YY, and pancreatic polypeptide (PP), the latter showing a preference for Y4R. A series cyclic oligopeptidic Y4R ligands were prepared applying novel approach, N-terminus to arginine side-chain cyclization. Most peptides acted as partial agonists, up 60-fold higher affinity compared linear precursor peptides. Two hexapeptides...
Within the family of neuropeptide Y (NPY) receptors, Y4 receptor (Y4R) is unique as it prefers pancreatic polypeptide over NPY and peptide YY. Today, low-molecular-weight Y4R ligands are lacking, in particular antagonists. We synthesized a series peptidic ligands, derived from hexapeptide acetyl-Arg-Tyr-Arg-Leu-Arg-Tyr-NH2 (1), reported to be partial agonist with high affinity (pKi Y4R: 8.43). Peptide 1 was N-terminally extended well truncated subjected d-amino acid scan, Leu replaced by...
Muscarinic acetylcholine receptors (MRs), comprising five subtypes (M1R–M5R) in humans, exhibit a high degree of structural similarity. Therefore, subtype-selective MR agonists and antagonists are lacking. We present an approach to highly M2R-selective based on the conjugation di- or tripeptides M2R-preferring dibenzodiazepinone-type antagonists. M2R selectivity was dependent peptide sequence type linker. The introduction basic amino acids resulted improved (e.g., UR-AP148 (48): pKi (hM2R)...
Abstract Histamine H 4 receptor (H R) orthologues are G‐protein‐coupled receptors (GPCRs) that exhibit species‐dependent basal activity. In contrast to the basally inactive mouse R (mH R), human (hH shows a high degree of We have performed long‐timescale molecular dynamics simulations and rigidity analyses on wild‐type hH R, experimentally characterized variants S179M, F169V, F169V+S179M, F168A, mH investigate nature differential variant‐dependent differences between essential motifs GPCR...
Abstract New classes of alkylated hetarylpropylguanidines with different functionality and variation in spacer length were synthesized to determine their behavior at the four histamine receptor (H 1 R, H 2 3 4 R) subtypes. Alkylated guanidines terminal functional groups varied basicity, like amine, guanidine urea developed, based on lead structure SK&F 91486 ( ). Furthermore, heteroatomic exchange led simple analogues compound. Radioassays all subtypes accomplished, as well organ bath...
The drug discovery process currently employed in the pharmaceutical industry typically requires about 10 years and $2-3 billion to deliver one new drug. This is both too expensive slow, especially emergencies like COVID-19 pandemic. In silicomethodologies need be improved better select lead compounds that can proceed later stages of protocol accelerating entire process. No single methodological approach achieve necessary accuracy with required efficiency. Here we describe multiple...
The recently resolved crystal structure of the neuropeptide Y Y1 receptor (Y1R), co-crystallized with high-affinity (pKi: 10.11), argininamide-type Y1R antagonist UR-MK299 (2), revealed that Nω-carbamoyl substituent (van der Waals volume: 139 Å3) is deeply buried in receptor, occupying a hydrophobic pocket. We synthesized and characterized series argininamides, structurally related to 2. affinity decreased increasing size carbamoyl residue (minimal pKi: 5.67). Exceeding critical 212 Å3),...