@phdthesis{He2024,
  author    = {He, Feng},
  title     = {Drug Discovery based on Oxidative Stress and HDAC6 for Treatment of Neurodegenerative Diseases},
  doi       = {10.25972/OPUS-25349},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-253497},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2024},
  abstract  = {Most antioxidants reported so far only achieved limited success in AD clinical trials. Growing evidences suggest that merely targeting oxidative stress will not be sufficient to fight AD. While multi-target directed ligands could synergistically modulate different steps in the neurodegenerative process, offering a promising potential for treatment of this complex disease. Fifteen target compounds have been designed by merging melatonin and ferulic acid into the cap group of a tertiary amide HDAC6 inhibitor. Compound 10b was screened as the best hybrid molecule exhibit potent HDAC6 inhibition and potent antioxidant capacity. Compound 10b also alleviated LPS-induced microglia inflammation and led to a switch from neurotoxic M1 to the neuroprotective M2 microglial phenotype. Moreover, compound 10b show pronounced attenuation of spatial working memory and long-term memory damage in an in vivo AD mouse model. Compound 10b can be a potentially effective drug candidate for treatment of AD and its druggability worth to be further studied. We have designed ten novel neuroprotectants by hybridizing with several common antioxidants, including ferulic acid, melatonin, lipoic acid, and trolox. The trolox hybrid compound exhibited the most potent neuroprotective effects in multiple neuroprotection assays. Besides, we identified the synergistic effects between trolox and vitamin K derivative, and our trolox hybrid compound showed comparable neuroprotection with the mixture of trolox and vitamin K derivative. We have designed and synthesized 24 quinone derivatives based on five kinds of different quinones including ubiquinone, 2,3,5-trimethyl-1,4-benzoquinone, memoquin, thymoquinone, and anthraquinone. Trimethylbenzoquinone and thymoquinone derivatives showed more potent neuroprotection than other quinones in oxytosis assay. Therefore, trimethylbenzoquinone and thymoquinone derivatives can be used as lead compounds for further mechanism study and drug discovery for treatment of neurodegenerative disease. We designed a series of photoswitchable HDAC inhibitors, which could be effective molecular tools due to the high spatial and temporal resolution. In total 23 target compounds were synthesized and photophysicochemically characterized. Azoquinoline-based compounds possess more thermally stable cis-isomers in buffer solution, which were further tested in enzyme-based HDAC inhibition assay. However, none of those tested compounds show significant differences in activities between trans-isomers and corresponding cis-isomers.},
  subject      = {Alzheimerkrankheit},
  language  = {en}
}
@phdthesis{Riemens2023,
  author    = {Riemens, Renzo J. M.},
  title     = {Neuroepigenomics in Alzheimer's disease: The single cell ADds},
  isbn      = {978-94-6423-524-1},
  doi       = {10.25972/OPUS-25457},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-254574},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2023},
  abstract  = {Die Forschung, die in dieser Arbeit zusammengestellt wird, kann in zwei Teile geteilt werden. Der erste Teil, bestehend aus vier Kapiteln, konzentriert sich auf die Rolle der epigenetischen Dysregulation in der {\"A}tiopathophysiologie der sporadischen Alzheimer-Krankheit (sAD). Neben Einblicken in die neuesten Entwicklungen in neuroepigenomischen Studien zu dieser Krankheit geht der erste Teil der Arbeit auch auf verbleibende Herausforderungen ein und gibt einen Ausblick auf m{\"o}gliche Entwicklungen auf diesem Gebiet. Der zweite Teil, der drei weitere Kapitel umfasst, konzentriert sich auf die Anwendung von auf induzierten pluripotenten Stammzellen (iPSC) basierenden Krankheitsmodellen f{\"u}r das Studium der AD, einschließlich, aber nicht beschr{\"a}nkt auf mechanistische Studien zur epigenetischen Dysregulation unter Verwendung dieser Plattform. Neben der Skizzierung der bisherigen Forschung mit iPSC-basierten Modellen f{\"u}r sAD gibt der zweite Teil der Arbeit auch Einblicke in die Gewinnung krankheitsrelevanter Nervenkulturen auf Basis der gezielten Differenzierung von iPSCs und beinhaltet dar{\"u}ber hinaus einen experimentellen Ansatz f{\"u}r den Aufbau eines solchen Modellsystems.},
  subject      = {Epigenetik},
  language  = {en}
}
@phdthesis{Hofmann2023,
  author    = {Hofmann, Julian},
  title     = {Synthesis of Sterubin, Flavonoid Hybrids, and Curcumin Bioisosteres and Characterization of their Neuroprotective Effects},
  doi       = {10.25972/OPUS-26664},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-266641},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2023},
  abstract  = {Alzheimer´s disease (AD) is a neurodegenerative disease and the most common form of dementia with still no preventive or curative treatment. Besides several risk factors, age is one of the major risks for AD and with an aging society, there is an urgent need for disease modifying agents. The strategy to address only one target within the intertwined network of AD failed so far. Natural products especially the phytochemical flavonoids, which are poly-phenolic natural products, have shown great potential as disease modifying agents against neurodegenerative disorders like Alzheimer´s disease (AD) with activities even in vivo. Flavonoids are produced by many plants and the native Californian plant Eriodictyon californicum is particularly rich in flavonoids. One of the major flavonoids of E. californicum is sterubin, a very potent agent against oxidative stress and inflammation, two hallmarks and drivers of AD and neurodegeneration. Herein, racemic sterubin was synthesized and separated into its pure (R)- and (S)-enantiomer by chiral HPLC. The pure enantiomers showed comparable neuroprotection in vitro with no significant differences. The stereoisomers were configurationally stable in methanol, but fast racemization was observed in culture medium. Moreover, the activity of sterubin was investigated in vivo, in an AD mouse model. Sterubin showed a significant positive impact on short- and long-term memory at low dosages. A promising concept for the increase of activity of single flavonoids is hybridization with aromatic acids like cinnamic or ferulic acids. Hybridization of the natural products taxifolin and silibinin with cinnamic acid led to an overadditive effect of these compounds in phenotypic screening assays related to neurodegeneration and AD. Because there are more potent agents as taxifolin or silibinin, the hybrids were further developed, and different flavonoid cinnamic acid hybrids were synthesized. The connection between flavonoids and cinnamic acid was achieved by an amide instead of a labile ester to improve the stability towards hydrolysis to gain better "druggability" of the compounds. To investigate the oxidation state of the C-ring of the flavonoid part, the dehydro analogues of the respective hybrids were also synthesized. The compounds show neuroprotection against oxytosis, ferroptosis and ATP-depletion in the murine hippocampal cell line HT22. While no overall trend within the flavanones compared to the flavones could be assigned, the taxifolin and the quercetin derivative were the most active compounds in course of all assays. The quercetin derivate even shows greater activity than the taxifolin derivate in every assay. As desired no hydrolysis product was found in cellular uptake experiments after 4h, whereas different metabolites were found. The last part of this work focused on synthetic bioisoteres of the natural product curcumin. Due to the drawbacks of curcumin and flavonoids arising from poor pharmacokinetics, rapid metabolism and sometimes instability in aqueous medium, we have examined the biological activity of azobenzene compounds designed as bioisoteres of curcumin, carrying the pharmacophoric catechol group of flavonoids. These bioisosteres exceeded their parent compounds in counteracting intracellular oxidative stress, neuroinflammation and amyloid-beta aggregation. By incorporating an azobenzene moiety and the isosteric behaviour to the natural parent compounds, these compounds may act as molecular tools for further investigation towards the molecular mode of action of natural products.},
  subject      = {Organische Synthese},
  language  = {en}
}
@phdthesis{Gunesch2021,
  author    = {Gunesch, Sandra},
  title     = {Molecular Mode of Action of Flavonoids: From Neuroprotective Hybrids to Molecular Probes for Chemical Proteomics},
  doi       = {10.25972/OPUS-23936},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-239360},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2021},
  abstract  = {Alzheimer's disease (AD) is the most common form of dementia, and currently, there is no treatment to cure or halt disease progression. Because the one-target strategy focusing on amyloid-β has failed to generate successful pharmaceutical treatment, this work studies natural products with pleiotropic effects focusing on oxidative stress and neuroinflammation as key drivers of disease progression. The central part of this work focused on flavonoids as neuroprotectants. 7-O-Esters of taxifolin and cinnamic or ferulic acid were synthesized and investigated towards their neuroprotective potential addressing aging and disease. 7-O-Feruloyl- and 7-O-cinnamoyltaxifolin showed overadditive effects in oxidative stress-induced assays in the mouse neuronal cell line HT22 and proved to be protective against neuroinflammation in microglial BV-2 cells. The overadditive effect translated to animals using an Aβ25-35-induced memory-impaired AD mouse model where the compounds were able to ameliorate short-term memory defects. While the disease-modifying effects in vivo were observed, the detailed mechanisms of action and intracellular targets of the compounds remained unclear. Hence, a chemical probe of the neuroprotective flavonoid ester 7-O-cinnamoyltaxifolin was developed and applied in an activity-based protein profiling approach. SERCA and ANT-1 were identified as potential targets. Further, chemical modifications on the flavonoids taxifolin, quercetin, and fisetin were performed. The achievements of this work are an important contribution to the use of secondary plant metabolites as neuroprotectants. Chemical modifications increased the neuroprotective effect of the natural products, and distinct intracellular pathways involved in the neuroprotective mechanisms were identified. The results of this work support the use of secondary plant metabolites as potential therapeutics and hint towards new pharmacological targets for the treatment of neurodegenerative disorders.},
  subject      = {Alzheimerkrankheit},
  language  = {en}
}
@phdthesis{Sawatzky2016,
  author    = {Sawatzky, Edgar},
  title     = {Design und Synthese selektiver Butyrylcholinesterase (BChE) Inhibitoren zur Entwicklung von Radiopharmazeutika zur Erforschung der Alzheimer Erkrankung},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-144037},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2016},
  abstract  = {Although the physiological roles of BChE are not yet determined to date, the importance of this enzyme is continuously increasing as it was found to be associated with several disorders like diabetes mellitus type 2, cardiovascular diseases, obesity and especially with Alzheimer's disease (AD). In consequence, for investigations of BChE's pathological role in these diseases and to find new medication strategies, the development of selective and potent inhibitors is necessary. For this purpose, the current work progresses in five chapters on the exploration of the chemical, physical and biochemical properties of tetrahydroquinazoline based carbamates which were previously reported to be selective BChE inhibitors with potency in the low nanomolar range. 1) A Novel Way to Radiolabel Human Butyrylcholinesterase for PET through Irreversible Transfer of the Radiolabeled Moiety: PET-radiotracers represent an innovative tool to determine the distribution and the expression of a biological target in vivo. BChE lacks to a large degree of such tracers with a few exceptions. In this work, methods were developed to incorporate the radioisotopes 11C and 18F into the carbamate moiety of an tetrahydroquinazoline based inhibitor. In contrast to reversibly acting PET-probes, the described radiotracers were proven by kinetic studies to transfer the radioisotope covalently onto the active site of BChE, thus labeling the enzyme directly and permanently. 2) Discovery of Highly Selective and Nanomolar Carbamate-Based Butyrylcholinesterase Inhibitors by Rational Investigation into Their Inhibition Mode: To investigate the role of the tetrahydroquinazoline carrier scaffold on BChE inhibition, carbamate based inhibitors were synthesized. These compounds were successively used to perform kinetic investigations to determine their inhibition mode. Based on these data, a plausible binding model was postulated explaining the influence of the tetrahydroquinazoline carrier scaffold for binding at BChE's active site just before carbamate transfer takes place. Additionally, these compounds feature neuroprotective properties and prevent oxidative stress induced cell death in their carbamate form as well as after the release of the tetrahydroquinazoline carrier scaffold. 3) Dual Addressing of Butyrylcholinesterase by Targeting the Catalytic Active Site (CAS) and the Peripheral Anionic Site (PAS): Compounds which are dual-targeting the CAS and the PAS of BChE are the most potent and selective BChE inhibitors to date with inhibition values in the picomolar range. In this work, a strategy is described how to turn tetrahydroquinazoline based carbamates into dual binding BChE inhibitors. These inhibitors feature a carbamate moiety which is covalently transferred onto the CAS of BChE, and in addition provide a second pharmacophore connected via a linker to the carbamate moiety which is proposed to target the PAS. Preliminary results reveal a high tolerance of BChE towards different linker lengths without decrease in affinity. 4) Investigation into Selective Debenzylation and Ring Cleavage of Quinazoline based Heterocycles: The tetrahydroquinazoline system is well investigated in terms of its synthesis and its selective oxidation. To explore the reactivity of this system, a tetracyclic tetrahydroquinazoline was exposed to common reduction agents. These experiments revealed a high sensitivity of the tetrahydroquinazoline core towards several reduction conditions 5) Experimental and Theoretical Investigation into the Stability of Cyclic Aminals: Tetrahydroquinazolines are known to degrade in acidic media through hydrolysis of their aminal system; but literature is lacking of a systematic investigation into this behavior. Therefore, different tetrahydroquinazolines were synthesized and exposed to phosphate buffered systems with defined pH-values. A clear increase of the hydrolysis rate of the aminal system was determined in dependency of an increasing acidic media. Computational studies predicted and experimental studies proved that hydrolysis takes place in an acidic environment while the condensation of this system is preferred in neutral or basic aqueous media.},
  subject      = {Cholinesterase},
  language  = {en}
}
@phdthesis{Sun2015,
  author    = {Sun, Ping},
  title     = {Alzheimer`s disease and brain insulin resistance: The diabetes inducing drug streptozotocin diminishes adult neurogenesis in the rat hippocampus - an in vivo and in vitro study},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-119252},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2015},
  abstract  = {Alzheimer's disease (AD) is the most prevalent neurodegenerative disease of the brain, which is characterized by a progressive loss of memory and spatial orientation. Only less than 5-10\% of AD sufferers are familial cases due to genetic mutations in the amyloid precursor protein (APP) gene or presenilin (PS) 1 and 2 genes. The cause of sporadic AD (sAD) which covers > 95\% of AD patients is still unknown. Current research found interactions between aging, diabetes and cognitive decline including dementia in general and in AD in particular. Disturbances of brain glucose uptake, glucose tolerance and utilization and impairment of the insulin/insulin receptor (IR) signaling cascade are thought to be key targets for the development of sAD. In the brain of AD patients, neural plasticity is impaired indicated by synaptic and neuronal loss. Adult neurogenesis (AN), the generation of functional neurons in the adult brain, may be able to restore neurological function deficits through the integration of newborn neurons into existing neural networks. The dentate gyrus of the hippocampus is one out of few brain regions where life-long AN exists. However, there is a big controversy in literature regarding the involvement of AN in AD pathology. Most animal studies used transgenic mice based on the Amyloid ß (Aß) hypothesis which primarily act as models for the familial form of AD. Findings from human post mortem AN studies were also inconstistent. In this thesis, we focused on the possible involvement of AN in the pathogenesis of the sporadic form of AD. Streptozotocin intracerebroventricularily (STZ icv) treated rats, which develop an insulin-resistant brain state and learning and memory deficits preceding Aß pathology act as an appropriate animal model for sAD. We used STZ treatment for both parts of my work, for the in vivo and in vitro study. In the first part of my thesis, my coworkers and I investigated STZ icv treatment effects on different stages of AN in an in vivo approach. Even if STZ icv treatment does not seem to considerably influence stem cell proliferation over a short-term (1 month after STZ icv treatment) as well as in a long-term (3 months after STZ icv treatment) period, it results in significantly less immature and newborn mature neurons 3 months after STZ icv treatment. This reduction detected after 3 months was specific for the septal hippocampus, discussed to be important for spatial learning. Subsequently we performed co-localization studies with antibodies detecting BrdU (applied appr. 27 days before sacrifice) and cell-type specific markers such as NeuN, and GFAP, we found that STZ treatment does not affect the differentiation fate of newly generated cells. Phenotype analysis of BrdU-positive cells in the hilus and molecular layer revealed that some of the BrdU-positive cells are newborn oligodendrocytes but not newborn microglia. In the second part of my thesis I worked with cultured neural stem cells (NSCs) isolated from the adult rat hippocampus to reveal STZ effects on the proliferation of of NSCs, and on the survival and differentiation of their progeny. Furthermore, this in vitro approach enabled me to study cellular mechanisms underlying the observed impaired neurogenesis in the hippocampus of STZ-treated rats. In contrast to our findings of the STZ icv in vivo study we revealed that STZ supplied with the cell culture medium inhibits the proliferation of NSCs in a dose-dependent and time-dependent manner. Moreover, performing immunofluorescence studies with antibodies detecting cell-type specific markers after triggering NSCs to differentiate, we could show that STZ treatment affects the number of newly generated neurons but not of astrocytes. Analyzing newborn cells starting to differentiate and migrate I was able to demonstrate that STZ has no effect on the migration of newborn cells. Trying to reveal cellular mechanisms underlying the negative influence of STZ on hippocampal AN, we performed qRT-PCR and immunofluorescence staining and thus could show that in NSCs the expression of glucose transporter (GLUT)3 mRNA as well as IR and GLUT3 protein levels are reduced after STZ treatment. Therefore, the inhibition of the proliferation of NSCs may be (at least partially) caused by these two molecules. Interestingly, the effect of STZ on differentiating cells was shown to be different, as IR protein expression was not significantly changed but GLUT3 protein levels were decreased in consequence of STZ treatment. In summary, this project delivered further insights into the interrelation between AN the sporadic form of sAD and thus provides a basis of new therapeutic approaches in sAD treatment through intervening AN. Discrepancies between the results of the two parts of my thesis, the in vivo and in vitro part, were certainly caused to a certain extent by the missing microenvironment in the in vitro approach with cultured NSCs. Future studies e.g. using co-culture systems could at least minimize the effect of a missing natural microenvironment of cultured NSCs, so that the use of an in vitro approach for the investigation of STZ treatment underlying cellular mechanisms can be improved.},
  subject      = {Alzheimerkrankheit},
  language  = {en}
}
@phdthesis{Kroker2011,
  author    = {Kroker, Katja},
  title     = {Establishment and validation of hippocampal LTP for characterization of memory enhancing drugs as potential treatment of Alzheimer's disease},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-85412},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2011},
  abstract  = {Die Alzheimer'sche Erkrankung ist eine neurodegenerative Erkrankung des Gehirns. Um geeignete Medikamente f{\"u}r die Behandlung der Alzheimer'schen Erkrankung zu finden, werden experimentelle Modellsysteme zur Erforschung von Substanzkandidaten verwendet. Ein solches experimentelles System ist die hippocampale Langzeitpotenzierung (LTP), welche ein anerkanntes in vitro Modell f{\"u}r die Erforschung der zugrundeliegenden zellul{\"a}ren Prozesse der Ged{\"a}chtnisbildung ist. Die vorliegende Arbeit besch{\"a}ftigt sich mit der Etablierung und Validierung von LTP in hippocampalen Hirnschnitten der Ratte um ged{\"a}chtnissteigernde Substanzen zur potentiellen Behandlung der Alzheimer'schen Erkrankung zu charakterisieren. Dazu wurde zun{\"a}chst ein Messsystem zur parallelen Charakterisierung mehrerer Schnitte aufgebaut, das Messungen bis zu sieben Stunden erlaubt (Kapitel 2). Dann wurden unterschiedliche Protokolle etabliert um Fr{\"u}h- und Sp{\"a}tphasen-LTP zu generieren. Dabei w{\"u}rde Fr{\"u}hphasen-LTP konzeptionell eher mit dem Kurzzeitged{\"a}chtnis einhergehen, w{\"a}hrend Sp{\"a}tphasen-LTP dem Langzeitged{\"a}chtnis gleichkommen w{\"u}rde (Kapitel 3). Da in Alzheimer-Patienten haupts{\"a}chlich ein Defizit cholinerger und glutamaterger Neurone vorliegt, wurden die validierten LTP Formen benutzt, um solche Substanzen zu analysieren, die potentiell cholinerge und/oder glutamaterge neuronale Funktion erh{\"o}hen. Die Effekte zweier ausschließlich cholinerge Funktion erh{\"o}hender Substanzen wurden analysiert: Der α4β2 nicotinische Acetylcholin-Rezeptor Agonist TC-1827 (Kapitel 4) und der Acetylcholinesterase-Inhibitor Donepezil (Kapitel 5). Beide Substanzen erh{\"o}hten Fr{\"u}hphasen-LTP, aber hatten keinen Effekt auf Sp{\"a}tphasen-LTP. Desweiteren wurden zwei Substanzen getestet, die ausschließlich mit glutamaterger Funktion interferieren: Der metabotrope Glutamatrezeptor 5 positiv allosterische Modulator ADX-47273 (Kapitel 3) und der Phosphodiesterase (PDE) 9A-Inhibitor BAY 73-6691 (Kapitel 5). ADX-47273 erh{\"o}hte Sp{\"a}tphasen-LTP, aber hatte keinen Effekt auf Fr{\"u}hphasen-LTP, wohingegen BAY 73-6691 eine erh{\"o}hende Wirkung auf beide LTP Formen aufwies und sogar Fr{\"u}h- in Sp{\"a}tphasen-LTP umwandelte. Die gleichen Effekte, wie bei dem PDE9A-Inhibitor, konnten auch mit dem partiellen α7 nicotinische Acetylcholin-Rezeptor Agonisten SSR180711 (Kapitel 4) demonstriert werden. SSR180711 wirkt sowohl auf cholinerge, als auch auf glutamaterge neuronale Funktion. Dann wurde die F{\"a}higkeit der Substanzen {\"u}berpr{\"u}ft, durch l{\"o}sliche Aβ Oligomere verschlechtertes LTP zu verbessern (Kapitel 6). L{\"o}sliche Aβ Oligomere, auch als amyloid-β derived diffusible ligands (ADDLs) bezeichnet, werden zurzeit als eine mutmaßliche Ursache der Alzheimer'schen Erkrankung angesehen. In der vorliegenden Arbeit wurde gezeigt, dass ADDLs Fr{\"u}h- und Sp{\"a}tphasen-LTP in verschiedenem Ausmaß vermindern. Donepezil und TC-1827 konnten die durch ADDLs induzierten Defizite bei Fr{\"u}hphasen-LTP geringf{\"u}gig wiederherstellen, aber sie hatten keinen Einfluss auf das durch ADDLs verschlechterte Sp{\"a}tphasen-LTP. Im Gegensatz dazu, konnten sowohl SSR180711 als auch BAY 73-6691 ein durch ADDLs verschlechtertes Fr{\"u}h- und Sp{\"a}tphasen-LTP komplett wiederherstellen. ADX-47273 hatte keinen positiven Effekt auf Fr{\"u}hphasen-LTP, welches durch ADDLs verschlechtert worden war, konnte aber ein durch ADDLs verschlechtertes Sp{\"a}tphasen-LTP teilweise wiederherstellen. Somit wurde der vorherige Befund der Arbeit best{\"a}tigt: Substanzen, welche die glutamaterge Funktion verbessern, scheinen nicht nur wirksamer im Bezug auf LTP-Erh{\"o}hung zu sein als Substanzen die ausschließlich cholinerge Funktion erh{\"o}hen, sondern sie sind auch in der Lage, durch l{\"o}sliche Aβ Oligomere verursachte Defizite bei LTP zu verbessern. Aus einem pr{\"a}klinischen Blickwinkel und basierend auf den Ergebnissen der vorliegenden Arbeit weisen demnach Substanzen, die glutamaterge Funktionen verbessern, ein hohes therapeutisches Potential als alternative Ans{\"a}tze bez{\"u}glich kognitiver Defizite auf. M{\"o}glicherweise k{\"o}nnten sie sogar wirksamere Ans{\"a}tze f{\"u}r die symptomatische Behandlung der Alzheimer'schen Erkrankung darstellen, als derzeitige Behandlungen, die ausschließlich cholinerge Funktion verbessern.},
  subject      = {Alzheimerkrankheit},
  language  = {en}
}