ABSTRACT
In this article, the earlier reported procedure for the synthesis of 2'-O-ß-D-ribofuranosyl nucleosides was extended to the synthesis of 2'-O-α-D-ribofuranosyl adenosine, a monomeric unit of poly(ADP-ribose). It consists in condensation of a small excess of 1-O-acetyl-2,3,5-tri-O-benzoyl-α,ß-D-arabinofuranose activated with tin tetrachloride with 3',5'-O-tetra-isopropyldisiloxane-1,3-diyl-ribonucleosides in 1,2-dichloroethane. The following debenzoylation and silylation of arabinofuranosyl residue and inversion of configuration at C-2'' atom of arabinofuranosyl residue and final removal of silyl protective groups gave 2'-O-α-D-ribofuranosyl adenosine in overall 13% to 21% yield. © 2019 by John Wiley & Sons, Inc.
Subject(s)
Adenosine/chemistry , Poly Adenosine Diphosphate Ribose/chemical synthesisABSTRACT
Poly(ADP-ribose) polymerase-1 (PARP-1) is an important target in cancer therapy. We present the synthesis of novel disaccharide nucleoside analogues that resemble the central motif of poly(ADP-ribose) and test their inhibitory effects on human PARP-1. Some compounds show inhibition of enzymatic activity in vitro and thus might be interesting for further investigations.
Subject(s)
Poly (ADP-Ribose) Polymerase-1/antagonists & inhibitors , Poly Adenosine Diphosphate Ribose/analogs & derivatives , Poly Adenosine Diphosphate Ribose/pharmacology , Poly(ADP-ribose) Polymerase Inhibitors/chemistry , Poly(ADP-ribose) Polymerase Inhibitors/pharmacology , Chemistry Techniques, Synthetic , Disaccharides/chemical synthesis , Disaccharides/chemistry , Disaccharides/pharmacology , Drug Discovery , Humans , Poly (ADP-Ribose) Polymerase-1/metabolism , Poly Adenosine Diphosphate Ribose/chemical synthesis , Poly Adenosine Diphosphate Ribose/chemistry , Poly(ADP-ribose) Polymerase Inhibitors/chemical synthesisABSTRACT
Poly(ADP-ribose) (PAR) is an important biopolymer, which is involved in various life processes such as DNA repair and replication, modulation of chromatin structure, transcription, cell differentiation, and in pathogenesis of various diseases such as cancer, diabetes, ischemia and inflammations. PAR is the most electronegative biopolymer and this property is essential for its binding with a wide range of proteins. Understanding of PAR functions in cell on molecular level requires chemical synthesis of regular PAR oligomers. Recently developed methodologies for chemical synthesis of PAR oligomers, will facilitate the study of various cellular processes, involving PAR.
Subject(s)
Drug Design , Enzyme Inhibitors/pharmacology , Poly Adenosine Diphosphate Ribose/pharmacology , Poly(ADP-ribose) Polymerases/metabolism , Dose-Response Relationship, Drug , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Humans , Molecular Structure , Poly Adenosine Diphosphate Ribose/chemical synthesis , Poly Adenosine Diphosphate Ribose/chemistry , Structure-Activity RelationshipABSTRACT
The synthesis of the core motif of branched poly(adenosine diphosphate ribose) (poly(ADPr)) is described, and structural analysis reasserted the proposed stereochemistry for branching. For the synthesis, a ribose trisaccharide was first constructed with only α-O-glycosidic linkages. Finally, the adenine nucleobase was introduced via a Vorbrüggen-type glycosylation reaction. The orthogonality of the selected protecting groups was demonstrated, allowing for the construction of branched poly(ADPr) oligomers in the near future.
Subject(s)
Poly Adenosine Diphosphate Ribose/chemical synthesis , Glycosylation , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Poly Adenosine Diphosphate Ribose/chemistryABSTRACT
Poly(ADP-ribose) (PAR) is a natural polymer, taking part in numerous important cellular processes. Several enzymes are involved in biosynthesis and degradation of PAR. One of them, poly(ADP-ribose)polymerase-1 (PARP-1) is considered to be a perspective target for the design of new drugs, affecting PAR metabolism. The structure of PAR was established by enzymatic hydrolysis and further analysis of the products, but total chemical synthesis of PAR hasn't been described yet. Several approaches have been developed on the way to chemical synthesis of this unique biopolymer.
Subject(s)
Biological Products , Biopolymers , Poly Adenosine Diphosphate Ribose/chemistry , Poly Adenosine Diphosphate Ribose/metabolism , Binding Sites , Biosynthetic Pathways , Clinical Trials as Topic , Humans , Molecular Structure , Poly Adenosine Diphosphate Ribose/chemical synthesis , Poly(ADP-ribose) Polymerase Inhibitors/chemistry , Poly(ADP-ribose) Polymerase Inhibitors/pharmacology , Poly(ADP-ribose) Polymerase Inhibitors/therapeutic use , Poly(ADP-ribose) Polymerases/chemistry , Poly(ADP-ribose) Polymerases/metabolism , Protein Binding , Static ElectricityABSTRACT
The post-translational modification of proteins that is known as adenosine diphosphate ribosylation (ADPr) regulates a wide variety of important biological processes, such as DNA-damage repair and cellular metabolism. This modification is also involved in carcinogenesis and the process of aging. Therefore, a better understanding of the function of ADP-ribosylation is crucial for the development of novel therapeutics. To facilitate the elucidation of the biology of ADPr, the availability of well-defined fragments of poly(ADP-ribose) is essential. Herein we report a solid-phase synthetic approach for the preparation of ADP-ribose oligomers of exactly defined length. The methodology is exemplified by the first reported synthesis of an ADP-ribose dimer and trimer.
Subject(s)
Adenosine Diphosphate Ribose/chemistry , Poly Adenosine Diphosphate Ribose/chemical synthesis , DNA Damage , DNA Repair , Magnetic Resonance Spectroscopy , Phosphorylation , Poly Adenosine Diphosphate Ribose/chemistry , Solid-Phase Synthesis TechniquesABSTRACT
Poly(ADP-ribose) is a significant nucleic acid polymer involved with diverse functions in eukaryotic cells, yet no structural information is available. A method for the synthesis of (13)C, (15)N-poly(ADP-ribose) (PAR) has been developed to allow characterization of the polymer using multidimensional nuclear magnetic resonance (NMR) spectroscopy. Successful integration of pentose phosphate, nicotinamide adenine dinucleotide biosynthesis, and cofactor recycling pathways with poly(ADP-ribose) polymerase-1 permitted labeling of PAR from (13)C-glucose and (13)C, (15)N-ATP in a single pot reaction. The scheme is efficient, yielding approximately 400 nmoles of purified PAR from 5 mumoles ATP, and the behavior of the synthetic PAR is similar to data from PAR synthesized by cell extracts. The resonances for (13)C, (15)N-PAR were unambiguously assigned, but the polymer appears to be devoid of inherent regular structure. PAR may form an ordered macromolecular structure when interacting with proteins, and due to the extensive involvement of PAR in cell function and disease, further studies of PAR structure will be required. The labeled PAR synthesis reported here will provide an essential tool for the future study of PAR-protein complexes.
Subject(s)
Poly Adenosine Diphosphate Ribose/chemistry , Poly Adenosine Diphosphate Ribose/chemical synthesis , Poly(ADP-ribose) Polymerases/chemistry , Adenosine Triphosphate/chemistry , Adenosine Triphosphate/metabolism , Carbon Isotopes , Glucose/chemistry , Glucose/metabolism , Isotope Labeling/methods , Niacinamide/chemistry , Niacinamide/metabolism , Nicotinamidase/chemistry , Nicotinamidase/metabolism , Nitrogen Isotopes , Nucleic Acid Conformation , Poly Adenosine Diphosphate Ribose/biosynthesis , Poly(ADP-ribose) Polymerases/metabolismABSTRACT
Using a poly(ADP-ribose) binding assay on protein blots we examined the ability of rat testis histone H1 variants to establish non-covalent interactions with the polymer. All the H1 variants bound ADP-ribose polymers; the binding was salt resistant and highly specific, occurring even in the presence of a large excess of competitor DNA. A comparison among the H1 variants showed that H1t has the highest affinity for poly(ADP-ribose). Long and branched poly(ADP-ribose) molecules were found to be preferentially involved in the interaction with the histone variants. The results further corroborate the concept that non-covalent interactions of poly(ADP-ribose) with target proteins may constitute an important mechanism to modulate chromatin structure.
Subject(s)
Histones/chemistry , Poly Adenosine Diphosphate Ribose/chemistry , DNA/chemistry , Electrophoresis, Polyacrylamide Gel , Endopeptidase K , Histones/classification , Poly Adenosine Diphosphate Ribose/chemical synthesisABSTRACT
The homopolymer of ADP-ribose, poly(ADP-ribose), was synthesized in vitro by liver nuclei from NAD. The protein-poly(ADP-ribose) adducts were isolated and, after base hydrolysis or proteolysis by proteinase K, the free polymers were separated from NAD, ADP-ribose, AMP and adenosine, and quantitatively determined by reversed-phase chromatography on an Ultrasphere ODS 5-micron column. Oxidation of the polymer by sodium periodate and labeling with 3H by borotritiation maintained the polymeric structure, but its modification was detectable by the chromatographic system employed.