Stefan Trentmann - Academia.edu (original) (raw)
Uploads
Papers by Stefan Trentmann
Exploiting Lipocalin Biochemistry For The Treatment Of Allergy And Asthma: Discovery And Characterization Of An Anti-IL-4RA Therapeutic
/ Poster Discussion Session / Tuesday, May 17/8:15 AM-10:45 AM / Room 503-504 C21 NEW TARGETS FOR... more / Poster Discussion Session / Tuesday, May 17/8:15 AM-10:45 AM / Room 503-504 C21 NEW TARGETS FOR ASTHMA THERAPY ... Exploiting Lipocalin Biochemistry For The Treatment Of Allergy And Asthma: Discovery And Characterization Of An Anti-Il-4ra Therapeutic
Angiogenesis, Dec 9, 2015
Human tear lipocalin (Tlc) was utilized as a protein scaffold to engineer an Anticalin that speci... more Human tear lipocalin (Tlc) was utilized as a protein scaffold to engineer an Anticalin that specifically binds and functionally blocks vascular endothelial growth factor A (VEGF-A), a pivotal inducer of physiological angiogenesis that also plays a crucial role in several neovascular diseases. Starting from a naive combinatorial library where residues that form the natural ligand-binding site of Tlc were randomized, followed by affinity maturation, the final Anticalin PRS-050 was selected to bind all major splice forms of VEGF-A with picomolar affinity. Moreover, this Anticalin cross-reacts with the murine ortholog. PRS-050 efficiently antagonizes the interaction between VEGF-A and its cellular receptors, and it inhibits VEGF-induced mitogenic signaling as well as proliferation of primary human endothelial cells with subnanomolar IC 50 values. Intravitreal administration of the Anticalin suppressed VEGF-induced blood-retinal barrier breakdown in a rabbit model. To allow lasting systemic neutralization of VEGF-A in vivo, the plasma half-life of the Anticalin was extended by site-directed PEGylation. The modified Anticalin efficiently blocked VEGF-mediated vascular permeability as well as growth of tumor xenografts in nude mice, concomitantly with reduction in microvessel density. In contrast to bevacizumab, the Anticalin did not trigger platelet aggregation and thrombosis in human FccRIIa transgenic mice, thus suggesting an improved safety profile. Since neutralization of VEGF-A activity is well known to exert beneficial effects in cancer and other neovascular diseases, including wet agerelated macular degeneration, this Anticalin offers a novel potent small protein antagonist for differentiated therapeutic intervention in oncology and ophthalmology.
Analysis of Arabidopsis cDNA that shows homology to the tomato E8 cDNA
Plant Molecular Biology, Oct 1, 1995
Journal of Bacteriology, Dec 1, 1989
inserting a A placMu bacteriophage into gene gimS encoding glucosamine 6-phosphate synthetase (GI... more inserting a A placMu bacteriophage into gene gimS encoding glucosamine 6-phosphate synthetase (GImS), the key enzyme of amino sugar biosynthesis, a nonreverting mutant of Escherichia coli K-12 that was strictly dependent on exogenous N-acetyl-D-glucosamine or D-glucosamine was generated. Analysis of suppressor mutations rendering the mutant independent of amino sugar supply revealed that the catabolic enzyme D-glucosamine-6-phosphate isomerase (deaminase), encoded by gene nagB of the nag operon, was able to fulfill anabolic functions in amino sugar biosynthesis. The suppressor mutants invariably expressed the isomerase constitutively as a result of mutations in nagR, the locus for the repressor of the nag regulon. Suppression was also possible by transformation of glmS mutants with high-copy-number plasmids expressing the gene nagB. Efficient suppression of the gimS lesion, however, required mutations in a second locus, termed glmX, which has been localized to 26.8 min on the standard E. coli K-12 map. Its possible function in nitrogen or cell wail metabolism is discussed.
British Journal of Pharmacology, Feb 23, 2018
Background and purpose Anaemia of chronic disease (ACD) has been linked to iron restricted erythr... more Background and purpose Anaemia of chronic disease (ACD) has been linked to iron restricted erythropoiesis imposed by high circulating levels of hepcidin, a 25 amino acid hepatocyte-derived peptide that controls systemic iron homeostasis. Here we report the engineering of the human lipocalin-derived, small protein-based Anticalin PRS-080 hepcidin antagonist with high affinity and selectivity. Experimental approach Anticalin-and hepcidin-specific pharmacokinetic/pharmacodynamic modelling was used to design and select the suitable drug candidate based on half life extension and duration of hepcidin suppression. Development of a novel free hepcidin assay enabled accurate analysis of bioactive hepcidin suppression and elucidation of the observed plasma iron levels after PRS-080-PEG30 administration in vivo. Key results PRS-080 had a hepcidin binding affinity of 0.07 nM and, after coupling to 30kD PEG (PRS-080-PEG30), a half life of 43h in cynomolgus monkeys. Dose-dependent iron mobilization and hepcidin suppression was observed after a single intravenous dose of PRS-080-PEG30 in cynomolgus monkeys. Importantly, in these animals Ligand ID INN IUPAC Hepcidin-25 5378 NA NA
Data from A Highly Potent and Specific MET Therapeutic Protein Antagonist with Both Ligand-Dependent and Ligand-Independent Activity
PDF - 53KB, Demonstration of benign immunogenicity profile of PRS-110 in ex vivo PBMC model
PDF - 31KB, Characterisation of cell line panel for MET and HGF expression
PDF - 136KB, Demonstration of PRS-110 dependent MET trafficking in HT29 cells
By inserting a A placMu bacteriophage into gene gimS encoding glucosamine 6-phosphate synthetase ... more By inserting a A placMu bacteriophage into gene gimS encoding glucosamine 6-phosphate synthetase (GImS), the key enzyme of amino sugar biosynthesis, a nonreverting mutant of Escherichia coli K-12 that was strictly dependent on exogenous N-acetyl-D-glucosamine or D-glucosamine was generated. Analysis of suppressor mutations rendering the mutant independent of amino sugar supply revealed that the catabolic enzyme D-glucosamine-6-phosphate isomerase (deaminase), encoded by gene nagB of the nag operon, was able to fulfill anabolic functions in amino sugar biosynthesis. The suppressor mutants invariably expressed the isomerase constitutively as a result of mutations in nagR, the locus for the repressor of the nag regulon. Suppression was also possible by transformation of glmS mutants with high-copy-number plasmids expressing the gene nagB. Efficient suppression of the gimS lesion, however, required mutations in a second locus, termed glmX, which has been localized to 26.8 min on the sta...
An il-4-r-alpha bindende tropfen-lipocalinmuteine
Molecular and General Genetics MGG, 1993
The En/Spm-encoded TNPA protein binds to 12-bp DNA sequence motifs that are present in the subter... more The En/Spm-encoded TNPA protein binds to 12-bp DNA sequence motifs that are present in the subtermini of the transposable element. DNA binding of TNPA to monomeric and dimeric forms of the binding motif was analyzed by gel retardation and cross-linking studies. A DNA binding domain at the N-terminal and a dimerization domain at the C-terminal portion of TNPA were localized using deletion derivatives of TNPA. These domains are novel since no apparent homology has been found in the data bases. The stoichiometry of the TNPA-DNA complexes was analyzed. A special complex is formed with a tail-to-tail dimeric DNA binding motif, most probably involving two DNA-bound TNPA molecules that interact via their dimerization domains. In redox reactions the requirement for one or two disulfide bonds for DNA binding of TNPA was shown. The implications of these findings for the excision mechanism of En/Spm are discussed.
British journal of pharmacology, 2018
Anaemia of chronic disease (ACD) has been linked to iron-restricted erythropoiesis imposed by hig... more Anaemia of chronic disease (ACD) has been linked to iron-restricted erythropoiesis imposed by high circulating levels of hepcidin, a 25 amino acid hepatocyte-derived peptide that controls systemic iron homeostasis. Here, we report the engineering of the human lipocalin-derived, small protein-based anticalin PRS-080 hepcidin antagonist with high affinity and selectivity. Anticalin- and hepcidin-specific pharmacokinetic (PK)/pharmacodynamic modelling (PD) was used to design and select the suitable drug candidate based on t extension and duration of hepcidin suppression. The development of a novel free hepcidin assay enabled accurate analysis of bioactive hepcidin suppression and elucidation of the observed plasma iron levels after PRS-080-PEG30 administration in vivo. PRS-080 had a hepcidin-binding affinity of 0.07 nM and, after coupling to 30 kD PEG (PRS-080-PEG30), a t of 43 h in cynomolgus monkeys. Dose-dependent iron mobilization and hepcidin suppression were observed after a sing...
Methods for Preventing or Treating Disorders by Increasing Bioavailability of Iron and Related Pharmaceutical Formulation
Muteins with Tear Lipocalin Having Affinity to Human C-Met Receptor Tyrosine Kinase and Methods for Obtaining the Same
Binding proteins for hepcidin
Exploiting Lipocalin Biochemistry For The Treatment Of Allergy And Asthma: Discovery And Characterization Of An Anti-IL-4RA Therapeutic
/ Poster Discussion Session / Tuesday, May 17/8:15 AM-10:45 AM / Room 503-504 C21 NEW TARGETS FOR... more / Poster Discussion Session / Tuesday, May 17/8:15 AM-10:45 AM / Room 503-504 C21 NEW TARGETS FOR ASTHMA THERAPY ... Exploiting Lipocalin Biochemistry For The Treatment Of Allergy And Asthma: Discovery And Characterization Of An Anti-Il-4ra Therapeutic
Angiogenesis, Dec 9, 2015
Human tear lipocalin (Tlc) was utilized as a protein scaffold to engineer an Anticalin that speci... more Human tear lipocalin (Tlc) was utilized as a protein scaffold to engineer an Anticalin that specifically binds and functionally blocks vascular endothelial growth factor A (VEGF-A), a pivotal inducer of physiological angiogenesis that also plays a crucial role in several neovascular diseases. Starting from a naive combinatorial library where residues that form the natural ligand-binding site of Tlc were randomized, followed by affinity maturation, the final Anticalin PRS-050 was selected to bind all major splice forms of VEGF-A with picomolar affinity. Moreover, this Anticalin cross-reacts with the murine ortholog. PRS-050 efficiently antagonizes the interaction between VEGF-A and its cellular receptors, and it inhibits VEGF-induced mitogenic signaling as well as proliferation of primary human endothelial cells with subnanomolar IC 50 values. Intravitreal administration of the Anticalin suppressed VEGF-induced blood-retinal barrier breakdown in a rabbit model. To allow lasting systemic neutralization of VEGF-A in vivo, the plasma half-life of the Anticalin was extended by site-directed PEGylation. The modified Anticalin efficiently blocked VEGF-mediated vascular permeability as well as growth of tumor xenografts in nude mice, concomitantly with reduction in microvessel density. In contrast to bevacizumab, the Anticalin did not trigger platelet aggregation and thrombosis in human FccRIIa transgenic mice, thus suggesting an improved safety profile. Since neutralization of VEGF-A activity is well known to exert beneficial effects in cancer and other neovascular diseases, including wet agerelated macular degeneration, this Anticalin offers a novel potent small protein antagonist for differentiated therapeutic intervention in oncology and ophthalmology.
Analysis of Arabidopsis cDNA that shows homology to the tomato E8 cDNA
Plant Molecular Biology, Oct 1, 1995
Journal of Bacteriology, Dec 1, 1989
inserting a A placMu bacteriophage into gene gimS encoding glucosamine 6-phosphate synthetase (GI... more inserting a A placMu bacteriophage into gene gimS encoding glucosamine 6-phosphate synthetase (GImS), the key enzyme of amino sugar biosynthesis, a nonreverting mutant of Escherichia coli K-12 that was strictly dependent on exogenous N-acetyl-D-glucosamine or D-glucosamine was generated. Analysis of suppressor mutations rendering the mutant independent of amino sugar supply revealed that the catabolic enzyme D-glucosamine-6-phosphate isomerase (deaminase), encoded by gene nagB of the nag operon, was able to fulfill anabolic functions in amino sugar biosynthesis. The suppressor mutants invariably expressed the isomerase constitutively as a result of mutations in nagR, the locus for the repressor of the nag regulon. Suppression was also possible by transformation of glmS mutants with high-copy-number plasmids expressing the gene nagB. Efficient suppression of the gimS lesion, however, required mutations in a second locus, termed glmX, which has been localized to 26.8 min on the standard E. coli K-12 map. Its possible function in nitrogen or cell wail metabolism is discussed.
British Journal of Pharmacology, Feb 23, 2018
Background and purpose Anaemia of chronic disease (ACD) has been linked to iron restricted erythr... more Background and purpose Anaemia of chronic disease (ACD) has been linked to iron restricted erythropoiesis imposed by high circulating levels of hepcidin, a 25 amino acid hepatocyte-derived peptide that controls systemic iron homeostasis. Here we report the engineering of the human lipocalin-derived, small protein-based Anticalin PRS-080 hepcidin antagonist with high affinity and selectivity. Experimental approach Anticalin-and hepcidin-specific pharmacokinetic/pharmacodynamic modelling was used to design and select the suitable drug candidate based on half life extension and duration of hepcidin suppression. Development of a novel free hepcidin assay enabled accurate analysis of bioactive hepcidin suppression and elucidation of the observed plasma iron levels after PRS-080-PEG30 administration in vivo. Key results PRS-080 had a hepcidin binding affinity of 0.07 nM and, after coupling to 30kD PEG (PRS-080-PEG30), a half life of 43h in cynomolgus monkeys. Dose-dependent iron mobilization and hepcidin suppression was observed after a single intravenous dose of PRS-080-PEG30 in cynomolgus monkeys. Importantly, in these animals Ligand ID INN IUPAC Hepcidin-25 5378 NA NA
Data from A Highly Potent and Specific MET Therapeutic Protein Antagonist with Both Ligand-Dependent and Ligand-Independent Activity
PDF - 53KB, Demonstration of benign immunogenicity profile of PRS-110 in ex vivo PBMC model
PDF - 31KB, Characterisation of cell line panel for MET and HGF expression
PDF - 136KB, Demonstration of PRS-110 dependent MET trafficking in HT29 cells
By inserting a A placMu bacteriophage into gene gimS encoding glucosamine 6-phosphate synthetase ... more By inserting a A placMu bacteriophage into gene gimS encoding glucosamine 6-phosphate synthetase (GImS), the key enzyme of amino sugar biosynthesis, a nonreverting mutant of Escherichia coli K-12 that was strictly dependent on exogenous N-acetyl-D-glucosamine or D-glucosamine was generated. Analysis of suppressor mutations rendering the mutant independent of amino sugar supply revealed that the catabolic enzyme D-glucosamine-6-phosphate isomerase (deaminase), encoded by gene nagB of the nag operon, was able to fulfill anabolic functions in amino sugar biosynthesis. The suppressor mutants invariably expressed the isomerase constitutively as a result of mutations in nagR, the locus for the repressor of the nag regulon. Suppression was also possible by transformation of glmS mutants with high-copy-number plasmids expressing the gene nagB. Efficient suppression of the gimS lesion, however, required mutations in a second locus, termed glmX, which has been localized to 26.8 min on the sta...
An il-4-r-alpha bindende tropfen-lipocalinmuteine
Molecular and General Genetics MGG, 1993
The En/Spm-encoded TNPA protein binds to 12-bp DNA sequence motifs that are present in the subter... more The En/Spm-encoded TNPA protein binds to 12-bp DNA sequence motifs that are present in the subtermini of the transposable element. DNA binding of TNPA to monomeric and dimeric forms of the binding motif was analyzed by gel retardation and cross-linking studies. A DNA binding domain at the N-terminal and a dimerization domain at the C-terminal portion of TNPA were localized using deletion derivatives of TNPA. These domains are novel since no apparent homology has been found in the data bases. The stoichiometry of the TNPA-DNA complexes was analyzed. A special complex is formed with a tail-to-tail dimeric DNA binding motif, most probably involving two DNA-bound TNPA molecules that interact via their dimerization domains. In redox reactions the requirement for one or two disulfide bonds for DNA binding of TNPA was shown. The implications of these findings for the excision mechanism of En/Spm are discussed.
British journal of pharmacology, 2018
Anaemia of chronic disease (ACD) has been linked to iron-restricted erythropoiesis imposed by hig... more Anaemia of chronic disease (ACD) has been linked to iron-restricted erythropoiesis imposed by high circulating levels of hepcidin, a 25 amino acid hepatocyte-derived peptide that controls systemic iron homeostasis. Here, we report the engineering of the human lipocalin-derived, small protein-based anticalin PRS-080 hepcidin antagonist with high affinity and selectivity. Anticalin- and hepcidin-specific pharmacokinetic (PK)/pharmacodynamic modelling (PD) was used to design and select the suitable drug candidate based on t extension and duration of hepcidin suppression. The development of a novel free hepcidin assay enabled accurate analysis of bioactive hepcidin suppression and elucidation of the observed plasma iron levels after PRS-080-PEG30 administration in vivo. PRS-080 had a hepcidin-binding affinity of 0.07 nM and, after coupling to 30 kD PEG (PRS-080-PEG30), a t of 43 h in cynomolgus monkeys. Dose-dependent iron mobilization and hepcidin suppression were observed after a sing...
Methods for Preventing or Treating Disorders by Increasing Bioavailability of Iron and Related Pharmaceutical Formulation
Muteins with Tear Lipocalin Having Affinity to Human C-Met Receptor Tyrosine Kinase and Methods for Obtaining the Same
Binding proteins for hepcidin