TY  - THES
A1  - Götz, Marcus Rudolf
T1  - Effiziente Synthese von Dronabinol und weiterer cannabinoider Derivate und deren pharmakologische Charakterisierung
T1  - An efficient synthesis of Dronabinol and further cannabinoid derivatives and their pharmacological characterization
N2  - In dieser Arbeit wurde ein Verfahren zur effizienten Herstellung von (−)-trans-Cannabidiol (CBD, 10), (−)-trans-Δ9-Tetrahydrocannabinol (Dronabinol, 21) und (−)-trans-Cannabidivarin (CBDV, 30) durch kontinuierliche Synthese untersucht und entwickelt. 
CBD konnte durch kontinuierliche Synthese in drei Schritten aus Olivetolcarbonsäuremethylester (OM, 6) und Menthadienol G (3) mit einer Ausbeute von 41 % synthetisiert werden. Bei optimierten Bedingungen betrug die Reinheit nach Kristallisation > 99 %. Die Stereochemie konnte durch Röntgenstrukturanalyse eindeutig als 1R,6R bestimmt werden. Vorteilhaft war dabei, dass Toluol anstatt eines chlorierten Lösungsmittels verwendet werden konnte. Weitere Vorteile waren die kurze Reaktionszeit und die Tatsache, dass die Synthese bei Raumtemperatur durchgeführt werden konnte. Es konnten fünf Nebenprodukte detektiert und identifiziert werden, wovon eines Dronabinol war.
Bei optimierten Reaktionsparametern konnte eine Ausbeute an Dronabinol von 64,5 % erreicht werden. Durch Simulated Moving Bed (SMB)-Chromatographie konnte Dronabinol kontinuierlich mit einem Gehalt von > 95 % hergestellt werden. Nach der Synthese waren vier Verunreinigungen detektierbar, und zwar Olivetol (17), CBD, Exo-Tetrahydrocannabinol (Exo-THC, 23) und Δ8-Tetrahydrocannabinol (Δ8-THC, 22). Durch die SMB-Aufreinigung konnten alle Verunreinigungen auf einen monographiekonformen (USP 37) Gehalt abgereichert werden. Nach der finalen destillativen Aufarbeitung trat eine noch nicht identifizierte Verunreinigung in einem Gehalt von ca. 0,4 Flächen-% auf.
CBDV konnte durch kontinuierliche Synthese in drei Schritten aus Divarincarbonsäuremethylester (DM, 25) und Menthadienol G synthetisiert werden. Die Ausbeute betrug ca. 30 %, die Reinheit nach Kristallisation > 99 %. Es konnten fünf Nebenprodukte detektiert werden, die im Rahmen dieser Arbeit nicht weiter charakterisiert wurden.
Der Syntheseweg bietet durch Modifikation der Seitengruppen an Position 6 (R1) und Position 5 (R2) der Alkylbenzol-Gruppe Zugang zu synthetischen Cannabinoiden mit einem CBD- oder CBDV-Grundgerüst. Es wurden neun neue Cannabinoide hergestellt: 2-Hydroxyethylcannabidiolat (2-HEC, 31), 2-Hydroxypentylcannabidiolat (2 HPC, 32), Glycerylcannabidiolat (GCBD, 33), Cyclohexylcannabidiolat (CHC, 34), Hexylcannabidiolat (HC, 35), N-Methylsulfonylcannabidiolat (NMSC, 36), 2 Hydroxyethylcannabidivarinolat (2-HECBDV, 37), Cyclohexylcannabidivarinolat (CHCBDV, 38) und Hexylcannabidivarinolat (HCBDV, 39).
Die Bindungsaffinität wurde in Cannabinoid-Rezeptor-transfizierten HEK293EBNA-Zellen untersucht, die intrinsische Aktivität in CHO-Zellen, die Induktion von NF-κB (nuclear factor kappa B) sowie von NFAT (nuclear factor of activated T cells) in Jurkat-T Zellen, die Induktion proinflammatorischer Zytokine und Chemokine (Interleukin(IL)-6, IL-1β, CC Chemokinligand 2' (CCL2) und Tumornekrosefaktor(TNF)-α) auf mRNA-Ebene in RAW264.7-Makrophagen und die Expression von proinflammatorischen Zytokinen (IL-1β, IL-6, IL-8, TNF-α) und Prostaglandin E2 (PGE2) auf Proteinebene in primären humanen Monozyten. 
Die CBD-Derivate zeigten eine höhere Selektivität für CB2-Rezeptoren. Die CBDV-Derivate HCBDV und CHCBDV zeigten eine spezifische Bindung an CB1- und CB2-Rezeptoren im nanomolaren Bereich. 2-HEC, 2-HPC, GCBD und NMSC wirkten als Agonisten an CB2- und als Antagonisten am CB1-Rezeptor. CHC band an CB1 und CB2 im submikromolaren Bereich und schien ein Agonist für beide Rezeptoren zu sein. 2- HECBD wirkte als Agonist auf CB2-Rezeptoren und als Antagonist auf CB1-Rezeptoren. In Jurkat-T Zellen hemmte NMSC dosisabhängig die Aktivität von NF-κB sowie von NFAT. 2-HEC, 2-HPC und GCBD hemmten die Expression von NFAT ebenfalls dosisabhängig. CHC und HC reduzierten dosisabhängig die Expression von IL-1β- und CCL2-mRNA in RAW264.7-Makrophagen. NMSC hemmte in geringeren Dosen IL-1β, CCL2 sowie TNF-α und induzierte in höheren Dosen einen starken Anstieg der IL-6-mRNA. In primären humanen Monozyten hemmten 2 HEC und GCBD konzentrationsabhängig die Synthese von IL-1β, IL-6 und TNF-α. 2-HPC hemmte dosisabhängig die Bildung von TNF-α und IL-6. HC verminderte dosisabhängig die Freisetzung von TNF-α und IL-6. NMSC steigerte die durch LPS erhöhte Freisetzung von IL-1β noch weiter, hemmte aber TNF-α, IL-8 und PGE2.
Die hier untersuchten CBD- und CBDV-Derivate sind geeignet, gezielt an Cannabinoid-Rezeptoren zu wirken. Einige der Derivate könnten als selektive CB2-Agonisten genutzt werden. Die Länge des aliphatischen Rests an R2 von CBD (Pentyl-Cannabinoiden) und CBDV (Propyl-Cannabinoiden) korrelierte nicht mit der Bindungsaffinität. Eine höhere Polarität an R1 (2-HECBDV > NMSC > GCBD > 2-HEC) schien demgegenüber die agonistische Aktivität an CB2 zu begünstigen. Um den Ergebnissen zur Beziehung zwischen Struktur und Wirkung noch mehr Bedeutung zu geben, wären weitere synthetische Derivate und deren Testung notwendig.
N2  - In this thesis a process for the efficient production of (−)-trans-cannabidiol (CBD, 10), (−)-trans-Δ9-tetrahydrocannabinol (dronabinol, 21) und (−)-trans-cannabidivarin (CBDV, 30) by means of continuous synthesis was researched and developed.
CBD was synthesized in three steps using a continuous synthesis process from olivetol-carboxylic acid methyl ester (OM, 6) and Menthadienol G (3) with a yield of 41%. With optimized conditions, the purity after crystallization was >99%. The stereochemistry was distinctly determined as 1R,6R by X-ray crystal analysis. Beneficial was the fact that toluene was used instead of a chlorinated solvent. Further advantages comprised a short reaction time and the fact that the synthesis can be carried out at room temperature. Five by-products were detected and identified, one of them being dronabinol.
With optimized reaction parameters, a yield of up to 64.5% of dronabinol was achieved. By simulated moving bed (SMB) chromatography, dronabinol was produced repeatedly with a purity of >95%. After the synthesis process, four impurities were detectable, namely olivetol (17), CBD, exo-tetrahydrocannabinol (exo-THC, 23) and Δ8-tetrahydrocannabinol (Δ8-THC, 22). All impurities were depleted to a monograph (USP 37) conforming level through SMB purification. After the final distillation, one further unidentified impurity occurred with a content of about 0.4 area%.
CBDV was synthesized through continuous synthesis in three steps from divarin-carboxylic acid methyl ester (DM, 25) und Menthadienol G. The yield was approximately 30%, the purity after crystallization >99%. Five by-products were detected which were not further characterized within the scope of this thesis.
Through modification of the side groups at position 6 (R1) and position 5 (R2) of the alkyl benzene moiety, the synthesis route offers access to synthetic cannabinoids with a CBD or CBDV scaffold. Nine new cannabinoids have been produced: 2-hydroxyethyl cannabidiolate (2-HEC, 31), 2-hydroxypentyl cannabidiolate (2-HPC, 32), glyceryl cannabidiolate (GCBD, 33), cyclohexyl cannabidiolate (CHC, 34), hexyl cannabidiolate (HC, 35), N-methyl-sulfonyl cannabidiolate (NMSC, 36), 2-hydroxyethyl cannabidivarinolate (2-HECBDV, 37), cyclohexyl cannabidivarinolate (CHCBDV, 38), hexyl cannabidivarinolate (HCBDV, 39). 
Binding affinity was studied in CB receptor-transfected HEK293EBNA cells, the intrinsic activity in CHO cells, the induction of nuclear factor kappa B (NF-κB) and nuclear factor of activated T cells (NFAT) in Jurkat T cells, the induction of pro-inflammatory cytokines and chemokines (interleukin (IL)-6, IL 1β, CC chemokine ligand 2 (CCL2) and tumor necrosis factor (TNF)-α) on the mRNA level in RAW264.7 macrophages and the expression of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, TNF-α) and prostaglandin E2 (PGE2) on the protein level in primary human monocytes.
The CBD derivatives showed higher selectivity for CB2. The CBDV derivatives HCBDV and CHCBDV showed specific binding at CB1 and CB2 receptors in the nanomolar range. 2-HEC, 2-HPC, GCBD and NMSC acted as agonists at CB2 and as antagonists at CB1 receptors. CHC bound CB1 and CB2 at the submicromolar range, and acted as an agonist for both receptors. 2-HECBDV was demonstrated to be an agonist at CB2 and an agonist at CB1. In Jurkat T cells, NMSC inhibited both NF-κB and NFAT activity in a dose-dependent fashion. 2-HEC, 2-HPC and GCBD inhibited the expression of NFAT, also in a dose-dependent manner. CHC and HC dose-dependently reduced the expression of IL-1β and CCL2 mRNA in RAW264.7 macrophages. NMSC inhibited at lower doses IL-1β, CCL2 and TNF-α and at higher doses induced a pronounced increase in IL-6 mRNA. In human primary monocytes, 2-HEC and GCBD inhibited IL-1β, IL-6 and TNF-α synthesis in a dose-dependent fashion. 2-HPC dose-dependently prevented the expression of TNF-α and IL-6 in high concentrations. HC decreased TNF-α and IL-6 release in a dose-dependent fashion. NMSC further increased LPS-elevated IL-1β release but inhibited TNF-α, IL-8 und PGE2. 
The CBD and CBDV derivatives studied here are suitable for targeting CB receptors. Some of the derivatives might be used as selective CB2 agonists. The length of the aliphatic rest at R2 of CBD (pentyl) and CBDV (propyl) did not correlate with binding affinity. Higher polarity at R1 (2-HECBDV > NMSC > GCBD > 2 HEC) however appeared to favor the agonistic activity at CB2 receptors. To give the results on the relationship between structure and effect more significance, further synthetic derivatives and their testing would be necessary.
KW  - Dronabinol
KW  - Cannabinoide
KW  - Cannabidiol
KW  - Cannabidivarin
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-166625
ER  - 
TY  - JOUR
A1  - Götz, Rudolf
A1  - Raulf, Friedrich
A1  - Schartl, Manfrad
T1  - Brain-derived neurotrophic factor is more highly conserved in structure and function than nerve growth factor during vertebrate evolution
N2  - Mammalian nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are members of a protein family with perfectly conserved domains arranged around the cysteine residues thought to stabilize an invariant three-dimensional scaffold in addition to distinct sequence motifs that convey different neuronal functions. To study their structural and functional conservation during evolution, we have compared NGF and BDNF from a lower vertebrate, the teleost fi.sh Xiphophorus, with the mammalian homlogues. Genomic clones encoding fish NGF and BDNF were isolated by cross-hybridization using probes from the cloned mammalian factors. Fish NGF and BDNF were expressed by means of recombinant vaccinia viruses, purified, and their neuronal survival specificities for different classes of neurons were found to mirror those of the mammalian factors. The half-maximal survival concentration for chick sensory neurons was 60 pg/ml for both fish and mammalian purifi.ed recombinant BDNF. However, the activity ofrecombinant fish NGF on both chick sensory and sympathetic neurons was 6 ng,lml, 75-fold lower than that of mouse NGF. The different functional conservation of NGF and BDNF is also reflected in their structures. The DNA-deduced amino acid sequences of processed mature fish NGF and BDNF showed, compared to mouse, 63% and 90% identity, respectively, indicating that NGF bad reached an optimized structure later than BDNF. The retrograde extrapolation of these data indicates that NGF and BDNF evolved at strikingly different rates ftom a common ancestral gene about 600 million years ago. By RNA gel blot anaJysis NGF mRNA was detected during late embryonie development; BDNF was present in adult brain.
KW  - Physiologische Chemie
KW  - Brain-derived neurotrophic factor
KW  - Nerve growth factor
KW  - Fish
KW  - Recombinant protein expression
KW  - Neuronal survival
Y1  - 1992
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61703
ER  - 
TY  - JOUR
A1  - Götz, Rudolf
A1  - Köster, Reinhard
A1  - Winkler, Christoph
A1  - Raulf, Friedrich
A1  - Lottspeich, Friedrich
A1  - Schartl, Manfred
A1  - Thoenen, Hans
T1  - Neurotrophin-6 is a new member of the nerve growth factor family
N2  - DURING vertebrale development, many neurons depend for survival and differentiation on their target cells\(^{1-3}\). The best documented mediator of such a retrograde trophic action is the neurotrophin nerve growth factor (NGF)\(^1\). NGF and the other known members of tbe neurotrophin family, brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT -3) and neurotrophin-4/5 (NT -4/5) are conserved as distinct genes over large evolutionary distances\(^{4 -6}\). Here we report the cloning of neurotrophin-6 (NT -6), a new member of this family from the teleost fish Xiphophorus. NT -6 distinguishes itself from the other known neurotrophins in that it is not found as a soluble protein in the medium of producing cells. The addition of heparin (but not chondroitin) effects the release of NT -6 from cell surface and extracellular matrix molecules. Recombinant purified NT -6 has a spectrum of actions similar to NGF on chick sympathetic and sensory neurons, albeit with a lower potency. NT -6 is expressed in tbe embryonie valvulla cerebelli; expression persists in some adult tissues. The interaction of NT-6 with heparin-binding molecuJes may modulate its action in the nervous system .
KW  - Physiologische Chemie
Y1  - 1994
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61544
ER  - 
TY  - JOUR
A1  - Götz, Rudolf
A1  - Sendtner, Michael
T1  - Cooperation of Tyrosine Kinase Receptor TrkB and Epidermal Growth Factor Receptor Signaling Enhances Migration and Dispersal of Lung Tumor Cells
JF  - PLoS ONE
N2  - TrkB mediates the effects of brain-derived neurotrophic factor (BDNF) in neuronal and nonnneuronal cells. Based on recent reports that TrkB can also be transactivated through epidermal growth-factor receptor (EGFR) signaling and thus regulates migration of early neurons, we investigated the role of TrkB in migration of lung tumor cells. Early metastasis remains a major challenge in the clinical management of non-small cell lung cancer (NSCLC). TrkB receptor signaling is associated with metastasis and poor patient prognosis in NSCLC. Expression of this receptor in A549 cells and in another adenocarcinoma cell line, NCI-H441, promoted enhanced migratory capacity in wound healing assays in the presence of the TrkB ligand BDNF. Furthermore, TrkB expression in A549 cells potentiated the stimulatory effect of EGF in wound healing and in Boyden chamber migration experiments. Consistent with a potential loss of cell polarity upon TrkB expression, cell dispersal and de-clustering was induced in A549 cells independently of exogeneous BDNF. Morphological transformation involved extensive cytoskeletal changes, reduced E-cadherin expression and suppression of E-cadherin expression on the cell surface in TrkB expressing tumor cells. This function depended on MEK and Akt kinase activity but was independent of Src. These data indicate that TrkB expression in lung adenoma cells is an early step in tumor cell dissemination, and thus could represent a target for therapy development.
KW  - metastasis
KW  - neurons
KW  - non-small cell lung cancer
KW  - neuron migration
KW  - adenocarcinoma of the lung
KW  - vector cloning
KW  - lung and intrathoracic tumors
KW  - secondary lung tumors
Y1  - 2014
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-119578
SN  - 1932-6203
VL  - 9
IS  - 6
ER  - 
TY  - JOUR
A1  - Zanucco, Emanuele
A1  - Götz, Rudolf
A1  - Potapenko, Tamara
A1  - Carraretto, Irene
A1  - Ceteci, Semra
A1  - Ceteci, Fatih
A1  - Seeger, Werner
A1  - Savai, Rajkumar
A1  - Rapp, Ulf R.
T1  - Expression of B-RAF V600E in Type II Pneumocytes Causes Abnormalities in Alveolar Formation, Airspace Enlargement and Tumor Formation in Mice
JF  - PLOS ONE
N2  - Growth factor induced signaling cascades are key regulatory elements in tissue development, maintenance and regeneration. Perturbations of these cascades have severe consequences, leading to developmental disorders and neoplastic diseases. As a major function in signal transduction, activating mutations in RAF family kinases are the cause of human tumorigenesis, where B-RAF V600E has been identified as the prevalent mutant. In order to address the oncogenic function of B-RAF V600E, we have generated transgenic mice expressing the activated oncogene specifically in lung alveolar epithelial type II cells. Constitutive expression of B-RAF V600E caused abnormalities in alveolar epithelium formation that led to airspace enlargements. These lung lesions showed signs of tissue remodeling and were often associated with chronic inflammation and low incidence of lung tumors. The inflammatory cell infiltration did not precede the formation of the lung lesions but was rather accompanied with late tumor development. These data support a model where the continuous regenerative process initiated by oncogenic B-RAF-driven alveolar disruption provides a tumor-promoting environment associated with chronic inflammation.
KW  - obstructive pulmonary-disease
KW  - lung-cancer
KW  - somatic mutations
KW  - epithelial-cells
KW  - mouse models
KW  - protein
KW  - kinase
KW  - inflammation
KW  - activation
KW  - pathway
Y1  - 2011
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-137061
VL  - 6
IS  - 12
ER  - 
TY  - JOUR
A1  - Pfeiffer, Verena
A1  - Götz, Rudolf
A1  - Xiang, Chaomei
A1  - Camarero, Guadelupe
A1  - Braun, Attila
A1  - Zhang, Yina
A1  - Blum, Robert
A1  - Heinsen, Helmut
A1  - Nieswandt, Bernhard
A1  - Rapp, Ulf R.
T1  - Ablation of BRaf Impairs Neuronal Differentiation in the Postnatal Hippocampus and Cerebellum
JF  - PLoS ONE
N2  - This study focuses on the role of the kinase BRaf in postnatal brain development. Mice expressing truncated, non-functional BRaf in neural stem cell-derived brain tissue demonstrate alterations in the cerebellum, with decreased sizes and fuzzy borders of the glomeruli in the granule cell layer. In addition we observed reduced numbers and misplaced ectopic Purkinje cells that showed an altered structure of their dendritic arborizations in the hippocampus, while the overall cornus ammonis architecture appeared to be unchanged. In male mice lacking BRaf in the hippocampus the size of the granule cell layer was normal at postnatal day 12 (P12) but diminished at P21, as compared to control littermates. This defect was caused by a reduced ability of dentate gyrus progenitor cells to differentiate into NeuN positive granule cell neurons. In vitro cell culture of P0/P1 hippocampal cells revealed that BRaf deficient cells were impaired in their ability to form microtubule-associated protein 2 positive neurons. Together with the alterations in behaviour, such as autoaggression and loss of balance fitness, these observations indicate that in the absence of BRaf all neuronal cellular structures develop, but neuronal circuits in the cerebellum and hippocampus are partially disturbed besides impaired neuronal generation in both structures.
KW  - granule cells
KW  - hippocampus
KW  - neurons
KW  - neuronal dendrites
KW  - embryos
KW  - dentate gyrus
KW  - neuronal differentiation
KW  - cerebellum
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130304
VL  - 8
IS  - 3
ER  - 
TY  - JOUR
A1  - Stöckli, K. A.
A1  - Lottspeich, F.
A1  - Sendtner, Michael
A1  - Masiakowski, P.
A1  - Carroll, Patrick
A1  - Götz, Rudolf
A1  - Lindholm, D.
A1  - Thoenen, Hans
T1  - Molecular cloning, expression and regional distribution of rat ciliary neurotrophic factor
N2  - CILIARY neurotrophic factor (CNTF) was originally characterized as a survival factor for chick ciliary neurons in vitro. More recently, it was shown to promote the survival of a variety of otherneuronal cell types and to affect the differentiation of E7 chick sympathetic neurons by inhibiting their proliferation and by inducing the expression of yasoactiYe intestinal peptide immunoreactiyity (VIP-IR). In cultures of dissociated sympathetic neurons from newborn rats, CNTF induces cholinergic differentiation as shown by increased levels of choline acetyltransferase (ChAT.
Y1  - 1989
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-34229
ER  -