RESUMEN
MET, the cell-surface receptor for the hepatocyte growth factor/scatter factor, which is widely overexpressed in various solid cancer types, is an attractive target for the development of antibody-based therapeutics. BYON3521 is a novel site-specifically conjugated duocarmycin-based antibody-drug conjugate (ADC), comprising a humanized cysteine-engineered IgG1 monoclonal antibody with low pmol/L binding affinity towards both human and cynomolgus MET. In vitro studies showed that BYON3521 internalizes efficiently upon MET binding and induces both target- and bystander-mediated cell killing. BYON3521 showed good potency and full efficacy in MET-amplified and high MET-expressing cancer cell lines; in moderate and low MET-expressing cancer cell lines good potencies and partial efficacy were observed. In mouse xenograft models, BYON3521 showed significant antitumor activity upon single-dose administration in multiple non-MET-amplified tumor types with low, moderate, and high MET expression, including complete tumor remissions in models with moderate MET expression. In the repeat-dose Good Laboratory Practice (GLP) safety assessment in cynomolgus monkeys, BYON3521 was well tolerated and based on the observed toxicities and their reversibility, the highest non-severely toxic dose was set at 15 mg/kg. A human pharmacokinetics (PK) model was derived from the PK data from the cynomolgus safety assessments, and the minimal efficacious dose in humans is estimated to be in the range of 3 to 4 mg/kg. In all, our nonclinical data suggests that BYON3521 is a safe ADC with potential for clinical benefit in patients. A first-in-human dose-escalation study is currently ongoing to determine the maximum tolerated dose and recommended dose for expansion (NCT05323045).
Asunto(s)
Anticuerpos Monoclonales , Inmunoconjugados , Animales , Humanos , Ratones , Anticuerpos Monoclonales Humanizados , Línea Celular Tumoral , Inmunoglobulina G , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Engineering cysteines at specific sites in antibodies to create well-defined ADCs for the treatment of cancer is a promising approach to increase the therapeutic index and helps to streamline the manufacturing process. Here, we report the development of an in silico screening procedure to select for optimal sites in an antibody to which a hydrophobic linker-drug can be conjugated. Sites were identified inside the cavity that is naturally present in the Fab part of the antibody. Conjugating a linker-drug to these sites demonstrated the ability of the antibody to shield the hydrophobic character of the linker-drug while resulting ADCs maintained their cytotoxic potency in vitro. Comparison of site-specific ADCs versus randomly conjugated ADCs in an in vivo xenograft model revealed improved efficacy and exposure. We also report a selective reducing agent that is able to reduce the engineered cysteines while leaving the interchain disulfides in the oxidized state. This enables us to manufacture site-specific ADCs without introducing impurities associated with the conventional reduction/oxidation procedure for site-specific conjugation.
Asunto(s)
Antibióticos Antineoplásicos/química , Cisteína/química , Duocarmicinas/análogos & derivados , Inmunoconjugados/química , Animales , Antibióticos Antineoplásicos/uso terapéutico , Línea Celular Tumoral , Duocarmicinas/uso terapéutico , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Inmunoconjugados/uso terapéutico , Inmunoglobulina G/química , Inmunoglobulina G/uso terapéutico , Ratones , Modelos Moleculares , Neoplasias/tratamiento farmacológico , Oxidación-ReducciónRESUMEN
Antibody-drug conjugates (ADCs) that are currently on the market or in clinical trials are predominantly based on two drug classes: auristatins and maytansinoids. Both are tubulin binders and block the cell in its progression through mitosis. We set out to develop a new class of linker-drugs based on duocarmycins, potent DNA-alkylating agents that are composed of a DNA-alkylating and a DNA-binding moiety and that bind into the minor groove of DNA. Linker-drugs were evaluated as ADCs by conjugation to the anti-HER2 antibody trastuzumab via reduced interchain disulfides. Duocarmycin 3b, bearing an imidazo[1,2-a]pyridine-based DNA-binding unit, was selected as the drug moiety, notably because of its rapid degradation in plasma. The drug was incorporated into the linker-drugs in its inactive prodrug form, seco-duocarmycin 3a. Linker attachment to the hydroxyl group in the DNA-alkylating moiety was favored over linking to the DNA-binding moiety, as the first approach gave more consistent results for in vitro cytotoxicity and generated ADCs with excellent human plasma stability. Linker-drug 2 was eventually selected based on the properties of the corresponding trastuzumab conjugate, SYD983, which had an average drug-to-antibody ratio (DAR) of about 2. SYD983 showed subnanomolar potencies against multiple human cancer cell lines, was highly efficacious in a BT-474 xenograft model, and had a long half-life in cynomolgus monkeys, in line with high stability in monkey and human plasma. Studies comparing ADCs with a different average DAR showed that a higher average DAR leads to increased efficacy but also to somewhat less favorable physicochemical and toxicological properties. Fractionation of SYD983 with hydrophobic interaction chromatography resulted in SYD985, consisting of about 95% DAR2 and DAR4 species in an approximate 2:1 ratio and having an average DAR of about 2.8. SYD985 combines several favorable properties from the unfractionated ADCs with an improved homogeneity. It was selected for further development and recently entered clinical Phase I evaluation.
Asunto(s)
Inmunoconjugados/química , Indoles/química , Receptor ErbB-2/metabolismo , Antineoplásicos/química , Antineoplásicos/farmacocinética , Línea Celular Tumoral , Duocarmicinas , Humanos , Inmunoconjugados/farmacocinética , Pirrolidinonas/químicaRESUMEN
We describe the synthesis of a series of interlocked structures from porphyrin-glycoluril cage compounds and bis(olefin)-terminated viologens by an olefin-metathesis protocol. The length of the chain connecting the olefin substituents with the viologen has a marked effect on the products of the ring-closure reaction. Long chains give [2]- and [3]catenane structures, whereas short chains give a mixture of [3]-, [4]-, and [5]catenanes. For comparison several [2]rotaxane compounds were prepared. The interlocked catenane and rotaxane structures display switching behavior, which can be controlled by the addition of acid and base. The kinetic and thermodynamic parameters of the switching processes have been determined by NMR spectroscopy.
RESUMEN
The cooperative binding effects of viologens and pyridines to a synthetic bivalent porphyrin receptor are used as a model system to study how the magnitudes of these effects relate to the experimentally obtained values. The full thermodynamic and kinetic circles concerning both activation and inhibition of the cage of the receptor for the binding of viologens were measured and evaluated. The results strongly emphasize the apparent character of measured binding and rate constants, in which the fractional saturation of receptors with other guests is linearly expressed in these constants. The presented method can be used as a simple tool to better analyze and comprehend the experimentally observed kinetics and thermodynamics of natural and artificial cooperative systems.
Asunto(s)
Porfirinas/química , Piridinas/química , Viológenos/química , Sitios de Unión , Cinética , Espectroscopía de Resonancia Magnética , Modelos Químicos , Modelos Moleculares , Estructura Molecular , TermodinámicaRESUMEN
The threading behavior of a zinc analogue of a previously reported processive manganese porphyrin catalyst onto a series of polymers of different lengths is reported. It is demonstrated that the speed of the threading process is determined by the opening of the cavity of the toroidal porphyrin host, which can be tuned with the help of axial ligands that coordinate to the metal center in the porphyrin.
Asunto(s)
Rotaxanos/química , Algoritmos , Catálisis , Cristalografía por Rayos X , Cinética , Ligandos , Luminiscencia , Conformación Molecular , Peso Molecular , Porfirinas/química , Termodinámica , Compuestos de Zinc/químicaRESUMEN
The kinetics and thermodynamics of the threading and dethreading process of polymers through the cavity of a synthetic toroidal host is investigated by studying its complexation with a series of end-functionalized polymers of different lengths containing an end group that is selectively recognized by the host. The system is designed in such a way that complexation is only observed if the host has traveled all of the way across the complete polymer. Detailed kinetic investigations using fluorescence spectroscopy have revealed that the barrier for this process is length dependent and most likely related to the stretching of the polymer. Moreover, the results indicate that our previously reported processive enzyme mimic most likely operates by randomly sliding along its macromolecular substrate.
Asunto(s)
Materiales Biomiméticos/química , Materiales Biomiméticos/metabolismo , Proteínas/química , Proteínas/metabolismo , Termodinámica , Cinética , Espectroscopía de Resonancia Magnética , Manganeso/química , Manganeso/metabolismo , Modelos Moleculares , Conformación Molecular , Peso Molecular , Oxidación-Reducción , Porfirinas/química , Porfirinas/metabolismo , Espectrometría de Fluorescencia , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Especificidad por SustratoRESUMEN
Proposed as intermediates in the catalytic oxidation of olefins to ketones, 3-rhoda-1,2-dioxolanes (κ2 C1 ,O2 -2-peroxyethyl rhodium complexes) have now been prepared by oxygenation of solid [(N4 -ligand)RhI (ethene)]PF6 with air. This process leads to stable isomeric 3-rhoda-1,2-dioxolanes A and B. Upon substitution of PF6- by BPh4- only isomer B is obtained. The X-ray structure of isomer B is presented.