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Chelate stabilization of a titanium(IV)–salan alkoxide by ligand exchange with 2,6-pyridinedicarboxylic acid (dipic) resulted in heptacoordinate complex 3 which is not redox-active, stable on silica gel and has increased aqueous stability. 3 is highly toxic in HeLa S3 and Hep G2 and has enhanced antitumor efficacy in a mouse cervical-cancer model.
Air- and water-stable phenyl complexes with nitridotechnetium(V) cores can be prepared by straightforward procedures. [TcNPh2(PPh3)2] is formed by the reaction of [TcNCl2(PPh3)2] with PhLi. The analogous N-heterocyclic carbene (NHC) compound [TcNPh2(HLPh)2], where HLPh is 1,3,4-triphenyl-1,2,4-triazol-5-ylidene, is available from (NBu4)[TcNCl4] and HLPh or its methoxo-protected form. The latter compound allows the comparison of different Tc–C bonds within one compound. Surprisingly, the Tc chemistry with such NHCs does not resemble that of corresponding Re complexes, where CH activation and orthometalation dominate.
N,N-Dialkylamino(thiocarbonyl)-N′-picolylbenzamidines react with (NEt4)2[M(CO)3X3] (M = Re, X = Br; M = Tc, X = Cl) under formation of neutral [M(CO)3L] complexes in high yields. The monoanionic NNS ligands bind in a facial coordination mode and can readily be modified at the (CS)NR1R2 moiety. The complexes [99Tc(CO)3(LPyMor)] and [Re(CO)3(L)] (L = LPyMor, LPyEt) were characterized by X-ray diffraction. Reactions of [99mTc(CO)3(H2O)3]+ with the N′-thiocarbamoylpicolylbenzamidines give the corresponding 99mTc complexes. The ester group in HLPyCOOEt allows linkage between biomolecules and the metal core.
Generation and Characterization of a Novel Multidrug Resistance Protein 2 Humanized Mouse Line
(2012)
Nico Scheer
; Praveen Balimane
; Michael D. Hayward
; Sandra Buechel
; Gunther Kauselmann
; C. Roland Wolf
The multidrug resistance protein (MRP) 2 is predominantly expressed in liver, intestine, and kidney, where it plays an important role in the excretion of a range of drugs and their metabolites or endogenous compounds into bile, feces, and urine. Mrp knockout [Mrp2(−/−)] mice have been used recently to study the role of MRP2 in drug disposition. Here, we describe the first generation and initial characterization of a mouse line humanized for MRP2 (huMRP2), which is nulled for the mouse Mrp2 gene and expresses the human transporter in the organs and cell types where MRP2 is normally expressed. Analysis of the mRNA expression for selected cytochrome P450 and transporter genes revealed no major changes in huMRP2 mice compared with wild-type controls. We show that human MRP2 is able to compensate functionally for the loss of the mouse transporter as demonstrated by comparable bilirubin levels in the humanized mice and wild-type controls, in contrast to the hyperbilirubinemia phenotype that is observed in MRP2(−/−) mice. The huMRP2 mouse provides a model to study the role of the human transporter in drug disposition and in assessing the in vivo consequences of inhibiting this transporter by compounds interacting with human MRP2.
Compared with rodents and many other animal species, the human cytochrome P450 (P450) Cyp2c gene cluster varies significantly in the multiplicity of functional genes and in the substrate specificity of its enzymes. As a consequence, the use of wild-type animal models to predict the role of human CYP2C enzymes in drug metabolism and drug-drug interactions is limited. Within the human CYP2C cluster CYP2C9 is of particular importance, because it is one of the most abundant P450 enzymes in human liver, and it is involved in the metabolism of a wide variety of important drugs and environmental chemicals. To investigate the in vivo functions of cytochrome P450 Cyp2c genes and to establish a model for studying the functions of CYP2C9 in vivo, we have generated a mouse model with a deletion of the murine Cyp2c gene cluster and a corresponding humanized model expressing CYP2C9 specifically in the liver. Despite the high number of functional genes in the mouse Cyp2c cluster and the reported roles of some of these proteins in different biological processes, mice deleted for Cyp2c genes were viable and fertile but showed certain phenotypic alterations in the liver. The expression of CYP2C9 in the liver also resulted in viable animals active in the metabolism and disposition of a number of CYP2C9 substrates. These mouse lines provide a powerful tool for studying the role of Cyp2c genes and of CYP2C9 in particular in drug disposition and as a factor in drug-drug interaction.
Harri Lempiäinen
; Philippe Couttet
; Federico Bolognani
; Arne Müller
; Valérie Dubost
; Raphaëlle Luisier
; Alberto del Rio-Espinola
; Veronique Vitry
; Elif B. Unterberger
; John P. Thomson
; Fridolin Treindl
; Ute Metzger
; Clemens Wrzodek
; Florian Hahne
; Tulipan Zollinger
; Sarah Brasa
; Magdalena Kalteis
; Magali Marcellin
; Fanny Giudicelli
; Albert Braeuning
; Laurent Morawiec
; Natasa Zamurovic
; Ulrich Längle
; Nico Scheer
; Dirk Schübeler
; Jay Goodman
; Salah-Dine Chibout
; Jennifer Marlowe
; Dietlinde Theil
; David J. Heard
; Olivier Grenet
; Andreas Zell
; Markus F. Templin
; Richard R. Meehan
; Roland C. Wolf
; Clifford R. Elcombe
; Michael Schwarz
; Pierre Moulin
; Rémi Terranova
; Jonathan G. Moggs
The molecular events during nongenotoxic carcinogenesis and their temporal order are poorly understood but thought to include long-lasting perturbations of gene expression. Here, we have investigated the temporal sequence of molecular and pathological perturbations at early stages of phenobarbital (PB) mediated liver tumor promotion in vivo. Molecular profiling (mRNA, microRNA [miRNA], DNA methylation, and proteins) of mouse liver during 13 weeks of PB treatment revealed progressive increases in hepatic expression of long noncoding RNAs and miRNAs originating from the Dlk1-Dio3 imprinted gene cluster, a locus that has recently been associated with stem cell pluripotency in mice and various neoplasms in humans. PB induction of the Dlk1-Dio3 cluster noncoding RNA (ncRNA) Meg3 was localized to glutamine synthetase-positive hypertrophic perivenous hepatocytes, sug- gesting a role for β-catenin signaling in the dysregulation of Dlk1-Dio3 ncRNAs. The carcinogenic relevance of Dlk1-Dio3 locus ncRNA induction was further supported by in vivo genetic dependence on constitutive androstane receptor and β-catenin pathways. Our data identify Dlk1-Dio3 ncRNAs as novel candidate early biomarkers for mouse liver tumor promotion and provide new opportunities for assessing the carcinogenic potential of novel compounds.
Nico Scheer
; Yury Kapelyukh
; Jillian McEwan
; Vincent Beuger
; Lesley A. Stanley
; Anja Rode
; C. Roland Wolf
The highly polymorphic human cytochrome P450 2D6 enzyme is involved in the metabolism of up to 25% of all marketed drugs and accounts for significant individual differences in response to CYP2D6 substrates. Because of the differences in the multiplicity and substrate specificity of CYP2D family members among species, it is difficult to predict pathways of human CYP2D6-dependent drug metabolism on the basis of animal studies. To create animal models that reflect the human situation more closely and that allow an in vivo assessment of the consequences of differential CYP2D6 drug metabolism, we have developed a novel straightforward approach to delete the entire murine Cyp2d gene cluster and replace it with allelic variants of human CYP2D6. By using this approach, we have generated mouse lines expressing the two frequent human protein isoforms CYP2D6.1 and CYP2D6.2 and an as yet undescribed variant of this enzyme, as well as a Cyp2d cluster knockout mouse. We demonstrate that the various transgenic mouse lines cover a wide spectrum of different human CYP2D6 metabolizer phenotypes. The novel humanization strategy described here provides a robust approach for the expression of different CYP2D6 allelic variants in transgenic mice and thus can help to evaluate potential CYP2D6-dependent interindividual differences in drug response in the context of personalized medicine.
Johannes Schiffels
; Olaf Pinkenburg
; Maximilian Schelden
; El-Hussiny A. A. Aboulnaga
; Marcus Baumann
; Thorsten Selmer
El-Hussiny Abulnaga
; Olaf Pinkenburg
; Johannes Schiffels
; Ahmed E-Refai
; Wolfgang Buckel
; Thorsten Selmer
Sandra Scheele
; Dan Oertel
; Johannes Bongaerts
; Stefan Evers
; Hendrik Hellmuth
; Karl-Heinz Maurer
; Michael Bott
; Roland Freudl
Anja Wilming
; Jens Begemann
; Stefan Kuhne
; Lars Regestein
; Johannes Bongaerts
; Stefan Evers
; Karl-Heinz Maurer
; Jochen Büchs
The response of Bacillus licheniformis to heat and ethanol stress and the role of the SigB regulon
(2013)
Birgit Voigt
; Rebecca Schroeter
; Britta Jürgen
; Dirk Albrecht
; Stefan Evers
; Johannes Bongaerts
; Karl-Heinz Maurer
; Thomas Schweder
; Michael Hecker
Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis
(2013)
Michael Rachinger
; Melanie Bauch
; Axel Strittmatter
; Johannes Bongaerts
; Stefan Evers
; Karl-Heinz Maurer
; Rolf Daniel
; Wolfgang Liebl
; Heiko Liesegang
; Armin Ehrenreich
Sandra Wiegand
; Sascha Dietrich
; Robert Hertel
; Johannes Bongaerts
; Stefan Evers
; Sonja Volland
; Rolf Daniel
; Heiko Liesegang
Shoko Takenaga
; Cony F. Herrera
; Frederik Werner
; Manfred Biselli
; Thomas Schnitzler
; Michael Josef Schöning
; Peter Öhlschläger
; Torsten Wagner