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Die vorliegende Erfindung betrifft neue Chinolon-Antibiotika und deren Verwendung. The present invention relates to novel quinolone antibiotics and to the use thereof.

Die vorliegende Erfindung betrifft neue Chinolon-Antibiotika und deren Verwendung, wie in den Ansprüchen definiert. The invention relates to novel quinolone antibiotics and to the use thereof, as defined in the claims.

Die vorliegende Erfindung betrifft neue Chinolon-Antibiotika und deren Verwendung, wie in den Ansprüchen definiert.

REACH (Registration, Evaluation, Authorisation and Restriction of Chemicals) is a regulation of the European Union, adopted to improve the protection of human health and the environment from risks that can be posed by chemicals, while enhancing the competitiveness of the EU chemical industry [1]. Often information on compound properties available to small companies is insufficient for REACH registration. Experimental testing of compounds is expensive and time consuming. Alternative methods for hazard assessment such as QSPR/QSAR (Quantitative structure ̶ property/activity relationships) are expressly recommended by regulatory authorities. However, there are at least two problems with this approach: i) Available computer programs for compound property prediction may be not appropriate for the particular compounds of interest. ii) Small chemical companies may not have personal sufficiently familiar with such programs to realistically estimate their limitations. Our project "Use of Cheminformatics models for the development and optimization of chemical products" is intended to alleviate this situation by supporting small companies in using appropriate computer programs. For example, a compound property of fundamental importance is biodegradability or not so. If a compound is readily biodegraded in the environment, there will be no major concern on bioaccumulation or long-term toxicity. Biodegradability is often predicted using the Biowin models of the U. S. EPA. However, the Biowin models are primarily based on the presence of certain substructures in molecules that are recognized by the program, for each such substructure a certain numerical contribution is added to a predicting function. If a compound structure is devoid of these substructures, and perhaps contains other substructures important for biodegradation, the Biowin predictions are based on the molecular mass only. A user unfamiliar with the programs may take such predictions literally, not realizing that his compounds are outside the domain of applicability of the programs. The approach will be to use a variety of biodegradability prediction software on particular classes of compounds containing less familiar functional groups, and to generate experimental biodegradation data for certain compounds with the aim to use these in establishing “local” biodegradation models for these compound classes in contrast to existing models that claim “global” applicability but were trained on compound classes containing the most fundamental functional groups only. This is a joint research project of Leuphana University Innovation Incubator and Bruno Bock GmbH, a leading producer of organosulfur compounds. Biodegradation tests for selected compounds will be performed according to OECD guidelines such as the Closed Bottle test (CBT). Reference:(1) http://echa.europa.eu/web/guest/regulations/reach/ (accessed May 13, 2014).

RATIONALE: The environmental fate of the organic pollutants such as drugs and their transformation products has received an increasing interest as a new research area over the last years. Psychiatric drugs are among the most important prescription drugs worldwide, therefore environmental fate and persistence of these pharmaceuticals and their degradation products is of high relevance and it is, yet, scarcely elucidated. Chlorprothixene (CPTX) is an antipsychotic drug of the thioxanthene class.METHODS: The photodegradation of CPTX was conducted using 150 W medium-pressures Hg-lamp and Xenon lamp in Ultra-pure water. The elimination process was continuously monitored to follow the transformation and mineralization rate of the parent compound using Liquid Chromatography combined with UV-Vis detection (LC–UV-Vis) and total organic carbon (TOC), respectively. Liquid chromatography coupled to ion-trap mass spectrometry (ESI–LC–MS/MS (ion-trap)) was employed to monitor and identify the newly formed photo-transformation products (PTPs). Biodegradation testing of CPTX itself and samples from photolysis was assessed employing standardized OECD tests: Closed Bottle test (CBT; OECD 301 D) and Manometric Respirometry test (MRT; OECD 301 F). Environmentally relevant properties and selected toxicity endpoints of CPTX and its PTPs were predicted using different QSAR software: Oasis Catalogic software V.5.11.6 TB, Case Ultra V 1.4.5.1, OECD QSAR toolbox V 3.2 and Leadscope software V.3.0.11-1.RESULTS: It was found that CPTX underwent complete transformation after 64 min with 13% mineralization under UV irradiation. The mineralization reached 33% after 128 min irradiation time. In case of xenon lamp, CPTX was partially transformed to its geometric isomer without any mineralization. Moreover, according to mass data, the putative molecular formulae for twenty different PTPs for CPTX could be established, most of them unprecedented. Thus, a pathway for the photo-transformation of CPTX in aqueous medium could be proposed. Neither the CPTX nor its PTPs were biodegradable in the OECD tests. There was a good correlation between the experimental findings for the PTPs and the modelled environmental relevant properties from the QSAR applications. Finally, in silico toxicity analysis of CPTX and its PTPs provided evidence for carcinogenicity, ecotoxicity and mutagenicity.CONCLUSIONS: These results indicate the importance of identification of the PTPs arising from photolysis treatment as these PTPs can be more hazardous to the environment than their parent compounds.

The antibiotic sulfamethoxypyridazine (SMP) undergoes phototransformation without complete mineralization. New phototransformation products (PTPs) are formed [1]. The photocatalytic degradation of SMP in ultrapure water (UP) and sewage treatment plant effluent (STP) was examined during the irradiation with a medium pressure Hg-lamp in the presence of TiO2, FeCl3 and TiO2/FeCl3 catalysts at natural pH (without using buffer or pH adjustment) in UP and STP effluent samples and pH3 in STP effluent samples. The primary elimination of SMP was monitored and structures of PTPs were assessed by LC-UV-MS/MS and total organic carbon (TOC) analyzer. The biodegradation of SMP samples after photolysis and photocatalysis was performed using Closed Bottle Test (CBT; OECD 301 D). Additional data for an environmental risk assessment of SMP and its PTPs were collected using in silico QSAR models.It was found that SMP underwent photocatalytic degradation and mineralization in the presence of TiO2, FeCl3 and TiO2/ FeCl3. The total degradation time decreased from 128 min in direct photolysis in UP to 16, 4, and 4 min in photocatalysis with TiO2, FeCl3 and TiO2/FeCl3, respectively and complete mineralization took place in case of TiO2 catalyst. In STP effluent, the degradation time decreased from about 64 min in direct photolysis to 32, 8, and 8 min in photocatalysis with TiO2, FeCl3 (pH3) and TiO2/FeCl3 (pH3), respectively. Based on the identified PTPs, a degradation pathway was proposed and a comparison between the three catalysts concerning the formed PTPs intensities was performed. On the one hand it was found that SMP and its PTPs were not readily biodegradable. On the other hand in HPLC-UV analysis, it was found that one of the PTPs showed full elimination at day 28 in CBT. QSAR results revealed that the formed PTPs might be more toxic than SMP to the environment with a higher risk of these PTPs on bacteria. Therefore, photocatalysis will be more preferable than photolysis as it offers complete mineralization. However the appropriate time period of irradiation has to be chosen.Acknowledgements: Waleed M. M. Mahmoud Ahmed thanks the Egyptian-German Research Long Term Scholarship (DAAD and MHESR Egypt) program for the scholarship. The authors wish to thank MultiCASE Inc. for providing CASE Ultra.[1] N. D. H. Khaleel, W. M. M. Mahmoud, G. M. Hadad, R. A. A. Salam, K. Kümmerer, Photolysis of sulfamethoxypyridazine in various aqueous media: aerobic biodegradation and identification of photoproducts by LC-UV-MS/MS. Journal of Hazardous Materials 2013 (244-245) 654-661.

Thalidomide (TD) became known as a sedative drug used for sleeping pills and morning sickness of pregnant women in the 1950s. In the 1960s, TD was withdrawn because of its teratogenicity. Recently, TD became a promising drug for the treatment of a number of cancers and inflammatory diseases. Consequently, a potential increased influx of TD into the aquatic environment has to be expected. Furthermore, incomplete degradation in surface water or effluent and drinking water treatment photolysis can occur. This makes it imperative to assess the fate and toxic effects of TD and its transformation products. In this study, the behavior of TD was monitored during irradiation with a medium-pressure Hg-lamp. The primary elimination of TD was monitored and structures of PTPs were assessed by LC-UV-FL-MS/MS. The estimation of the relevant properties of TD and its photoproducts (PTPs) and hydrolysis products (HTPs) was performed using in silico QSAR models. Mutagenicity of 47mg/L TD after 2, 4, 8, 16, 32, 64 and 128 min of irradiation was assessed in the Ames microplate format (MPF) aqua assay (Xenometrix, AG). In addition, toxicity towards environmental bacteria was investigated in a modified luminescent bacteria test using Vibrio fischeri. New PTPs were formed during irradiation. They were more polar than TD. All the PTPs peaks increased with irradiation time until 32 min and then began to decrease. One PTPs peak was formed at 16 min then increased until 128 min. Two of the PTPs were isomers of TD with the same molecular mass. TD and its PTPs did not exhibit mutagenic activities in the Salmonella typhimurium strains TA 98, and TA 100 with and without metabolic activation. In contrast, QSAR analysis of PTPs and HTPs provided evidence for mutagenicity and carcinogenicity by investigating additional endpoints in silico compared with the experimental Ames tests. In the luminescent bacteria test, a steady increase of toxicity during the treatment procedure was observed. The acute toxicity started to increase significantly after 16 min of irradiation compared to the parent compound. After 128min of irradiation, a maximum of 96% acute luminescence inhibition of the reaction mixtures was measured [dilution 1:2 (v/v)]. The diluted reaction mixtures [dilution 1:50 (v/v)] showed no significant inhibition. The analysis of luminescence inhibition and growth inhibition showed a significant toxification starting from 32 min of irradiation. QSAR analysis with these endpoints provided evidence for positive alerts for acute toxicity on environmental bacteria in several identified PTPs and HTPs.In conclusion, the UV irradiation eliminated TD itself but lead to the formation of several PTPs. Some of the PTPs are more toxic to the investigated bacteria than the parent compound but do not show mutagenic activity in the Ames test. Nevertheless the results show the toxic potential of the photo PTPs and deserve further attention.

Die vorliegende Erfindung betrifft ein Fusionsprotein, enthaltend a) ein erstes Polypeptid, das ausgewählt ist aus SDF-1 (Stromal Cell derived Factor-1) oder Peptidase-/Protease resistenten Varianten oder Fragmenten davon, die die CXCR4-/CXCR7-Bindungsfunktion von SDF-1 besitzen; und b) ein zweites Polypeptid, das ausgewählt ist aus GPVI (Glycoprotein VI), oder der extrazellulären Domäne von GPVI, oder Fragmenten oder Varianten der extrazellulären Domäne von GPVI, die die Kollagen-Bindungsfunktion von GPVI besitzen, wobei das erste Polypeptid und das zweite Peptid direkt oder über ein Linkermolekül miteinander verknüpft sind. Die Erfindung betrifft ferner die Verwendung des Fusionsproteins zur Behandlung von Erkrankungen.

The co-inhibitory B7-homologue 1 (B7-H1/PD-L1) influences adaptive immune responses and has been proposed to contribute to the mechanisms maintaining peripheral tolerance and limiting inflammatory damage in parenchymal organs. To understand the B7-H1/PD1 pathway in CNS inflammation, we analyzed adaptive immune responses in myelin oligodendrocyte glycoprotein (MOG)(35-55)-induced EAE and assessed the expression of B7-H1 in human CNS tissue. B7-H1(-/-) mice exhibited an accelerated disease onset and significantly exacerbated EAE severity, although absence of B7-H1 had no influence on MOG antibody production. Peripheral MOG-specific IFN-gamma/IL-17 T cell responses occurred earlier and enhanced in B7-H1(-/-) mice, but ceased more rapidly. In the CNS, however, significantly higher numbers of activated neuroantigen-specific T cells persisted during all stages of EAE. Experiments showing a direct inhibitory role of APC-derived B7-H1 on the activation of MOG-specific effector cells support the assumption that parenchymal B7-H1 is pivotal for delineating T cell fate in the target organ. Compatible with this concept, our data investigating human brain tissue specimens show a strong up-regulation of B7-H1 in lesions of multiple sclerosis. Our findings demonstrate the critical importance of B7-H1 as an immune-inhibitory molecule capable of down-regulating T cell responses thus contributing to the confinement of immunopathological damage. The co-inhibitory B7-homologue 1 (B7-H1/PD-L1) influences adaptive immune responses and has been proposed to contribute to the mechanisms maintaining peripheral tolerance and limiting inflammatory damage in parenchymal organs. To understand the B7-H1/PD1 pathway in CNS inflammation, we analyzed adaptive immune responses in myelin oligodendrocyte glycoprotein (MOG)(35-55)-induced EAE and assessed the expression of B7-H1 in human CNS tissue. B7-H1(-/-) mice exhibited an accelerated disease onset and significantly exacerbated EAE severity, although absence of B7-H1 had no influence on MOG antibody production. Peripheral MOG-specific IFN-gamma/IL-17 T cell responses occurred earlier and enhanced in B7-H1(-/-) mice, but ceased more rapidly. In the CNS, however, significantly higher numbers of activated neuroantigen-specific T cells persisted during all stages of EAE. Experiments showing a direct inhibitory role of APC-derived B7-H1 on the activation of MOG-specific effector cells support the assumption that parenchymal B7-H1 is pivotal for delineating T cell fate in the target organ. Compatible with this concept, our data investigating human brain tissue specimens show a strong up-regulation of B7-H1 in lesions of multiple sclerosis. Our findings demonstrate the critical importance of B7-H1 as an immune-inhibitory molecule capable of down-regulating T cell responses thus contributing to the confinement of immunopathological damage.

An object of the invention is to provide methods and compositions relating to a vaccine against prostate cancer which includes a non- human -primate PSA for administration to humans to provide an immune response against human PSA. More generally, an object of the invention is to provide methods and compositions relating to using a non-human primate xenogeneic antigen (e. g. protein) in a human, wherein, with respect to the non-human primate xenogeneic antigen that is used, there are relatively few interspecies differences between the non-human primate xenogeneic antigen and the human self antigen in order to induce an optimal immune response in the human to its native self antigen.

The causal interrelation of CNS degeneration and adaptive immunityis assumed as a critical pathogenic element influencing the clinicalphenotype in neuroinflammatory as well as in neurodegenerativedisorders. We have recently shown that primary oligodendrocyticdamage (by overexpression of PLP) leads to a low-grade inflammatoryreaction of high pathological impact in the CNS, potentially reflectingpathogenic principles in some forms of multiple sclerosis. Asa completely novel finding, these reactions were mediated by CD8+T cells whereas CD4+ cells were not necessary for the pathologicaleffects of the immune system. In order to yield further insights intopathogenic relevance and nature of the adaptive immune responsein myelin mutated mice (PLP transgenic mice), we here apply a detailedimmunological characterization of systemic and CNS-specificCD8+ T cell responses in PLP transgenic mice and aged wild typemice. We provide evidence that T effector cells are surveying the CNSof PLP transgenic and wild-type mice. These cells accumulate withage and show a higher level of activation in PLP transgenic mice.Exclusively in PLP transgenic mice, T cells show clonal expansions,as demonstrated by T cell receptor CDR3-spectratype analysis. Althoughwe could not find specific responses of CD8+ T cells againstmyelin-antigen-derived peptides, V-beta/J-beta similarities stronglysuggest specificity against a common antigen. Our data suggest thata primary degenerative CNS disorder, exemplified by PLP-transgenicmice, can be associated with secondary expansions of pathogenicallyrelevant CD8+ T cells in the CNS. These findings have implicationsfor our understanding of the role of secondary adaptive immunereactions in the context of neurodegenerative and neuroinflammatorydiseases.

Infections by human papillomaviruses (HPV) are the major cause of uterine cancer in women worldwide. Aiming to develop a combined prophylactic and therapeutic vaccine we have previously demonstrated immunogenicity of chimeric virus-like particles consisting of a C-terminally truncated HPV 16 L1 capsid protein fused to an E7 portion. Here we show that genetic vaccination with a corresponding DNA was inefficient in the induction of a L1-specific prophylactic immune response. DNA immunization with C-terminally truncated HPV 16 L1 genes of different lengths revealed that only short deletions (L1(1-498)) were tolerated for eliciting a humoral immune response against viral capsids. This correlates with the observation that the C-terminal sequences are critical for nuclear localization, capsomere and capsid assembly. However, only the ability of L1 protein to form capsomeres or capsids showed a direct influence on the outcome of the immune response. C-terminal insertion of 60 amino acids of E7 was tolerated in fusion constructs, whereas insertion of full-length E7(1-98) or shuffled E7 (149 aa) completely abolished the humoral immune response. The L1(1-498)/E7(1-60) fusion construct not only induced L1-specific antibodies but also L1- and E7-specific CTL responses after DNA vaccination.

In recent years, a number of tumor vaccination strategies have been developed. Most of these rely on the identification of tumor antigens that can be recognized by the immune system. DNA vaccination represents one such approach for the induction of both humoral and cellular immune responses against tumor antigens. Studies in animal models have demonstrated the feasibility of utilizing DNA vaccination to elicit protective antitumor immune responses. However, most tumor antigens expressed by cancer cells in humans are weakly immunogenic, and therefore require the development of strategies to potentiate DNA vaccine efficacy in the clinical setting. This review focuses on recent advances in understanding of the immunology of DNA vaccines, as well as strategies used to increase DNA vaccine potency with respect to cytotoxic T-lymphocyte activity. In recent years, a number of tumor vaccination strategies have been developed. Most of these rely on the identification of tumor antigens that can be recognized by the immune system. DNA vaccination represents one such approach for the induction of both humoral and cellular immune responses against tumor antigens. Studies in animal models have demonstrated the feasibility of utilizing DNA vaccination to elicit protective antitumor immune responses. However, most tumor antigens expressed by cancer cells in humans are weakly immunogenic, and therefore require the development of strategies to potentiate DNA vaccine efficacy in the clinical setting. This review focuses on recent advances in understanding of the immunology of DNA vaccines, as well as strategies used to increase DNA vaccine potency with respect to cytotoxic T-lymphocyte activity.

Described is a method of producing a repertoire of random peptides on the surface of AAV particles wherein said random peptides are expressed as a fusion with an AAV capsid protein of an AAV particle which displays at its surface said random polypeptides. Also described is a peptide library obtainable by said method as well as a method of selecting a gene therapy vector specific for a desired cell type comprising the steps of (a) infecting the desired cell type with a peptide library of the invention and (b) harvesting AAV library particles from the supernatant and/or cell lysates. Finally, AAV vectors obtained by said method are described which are useful for gene therapy, e.g., AAV vectors targeting primary human coronary artery endothelial cells which are suitable for the treatment of diseases associated with a dysfunction of said cells.

Infection of cells with adeno-associated virus (AAV) type 2 (AAV-2) is mediated by binding to heparan sulfate proteoglycan and can be competed by heparin. Mutational analysis of AAV-2 capsid proteins showed that a group of basic amino acids (arginines 484, 487, 585, and 588 and lysine 532) contribute to heparin and HeLa cell binding. These amino acids are positioned in three clusters at the threefold spike region of the AAV-2 capsid. According to the recently resolved atomic structure for AAV-2, arginines 484 and 487 and lysine 532 on one site and arginines 585 and 588 on the other site belong to different capsid protein subunits. These data suggest that the formation of the heparin-binding motifs depends on the correct assembly of VP trimers or even of capsids. In contrast, arginine 475, which also strongly reduces heparin binding as well as viral infectivity upon mutation to alanine, is located inside the capsid structure at the border of adjacent VP subunits and most likely influences heparin binding indirectly by disturbing correct subunit assembly. Computer simulation of heparin docking to the AAV-2 capsid suggests that heparin associates with the three basic clusters along a channel-like cavity flanked by the basic amino acids. With few exceptions, mutant infectivities correlated with their heparin- and cell-binding properties. The tissue distribution in mice of recombinant AAV-2 mutated in R484 and R585 indicated markedly reduced infection of the liver, compared to infection with wild-type recombinant AAV, but continued infection of the heart. These results suggest that although heparin binding influences the infectivity of AAV-2, it seems not to be necessary.