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1.
Nucleic Acids Res ; 49(15): 8462-8470, 2021 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-34358308

RESUMO

Small-molecules interacting with particular RNAs and modulating their functions are vital tools for RNA-targeting drug discovery. Considering the substantial distribution of the internal loops involving two contiguous cytosines opposite to a single-nucleotide base (Y/CC; Y = C, U or A) within the biologically significant functional RNAs, developing small-molecule probes targeting Y/CC sites should provide profound insight into their functions and roles in biochemical processes. Herein, we report ANP77 as the small-molecule probe for sensing RNA internal loop of Y/CC motifs and molecules binding to the motifs. The Y/CC motifs interact with ANP77 via the formation of a 1:1 complex and quench the fluorescence of ANP77. The flanking sequence-dependent binding to C/CC and U/CC sites was assessed by fluorometric screening, provided the binding heat maps. The quenching phenomena of ANP77 fluorescence was confirmed with intrinsic potential drug target pre-miR-1908. Finally, the binding-dependent fluorescence quenching of ANP77 was utilized in the fluorescence indicator displacement assay to demonstrate the potential of ANP77 as an indicator by using the RNA-binding drugs, risdiplam and branaplam.


Assuntos
Corantes Fluorescentes/química , RNA/química , Compostos Azo/metabolismo , Citosina/química , Descoberta de Drogas , MicroRNAs/química , Motivos de Nucleotídeos , Pirimidinas/metabolismo , RNA/metabolismo
2.
J Chem Phys ; 155(4): 044110, 2021 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-34340364

RESUMO

A multi-level layered elongation method was developed for efficiently analyzing the electronic states of local structures in large bio/nano-systems at the full ab initio level of theory. The original elongation method developed during the last three decades in our group has focused on the system in one direction from one terminal to the other terminal to sequentially construct the electronic states of a polymer, called a theoretical synthesis of polymers. In this study, an important region termed the central (C) part is targeted in a large polymer and the remainder are terminal (T) parts. The electronic structures along with polymer elongation are calculated repeatedly from both end T parts to the C central part at the same time. The important C part is treated with large basis sets (high level) and the other regions are treated with small basis sets (low level) in the ab initio theoretical framework. The electronic structures besides the C part can be reused for other systems with different structures at the C part, which renders the method computationally efficient. This multi-level layered elongation method was applied to the investigation on DNA single bulge recognition of small molecules (ligands). The reliability and validity of our approach were examined in comparison with the results obtained by direct calculations using a conventional quantum chemical method for the entire system. Furthermore, stabilization energies by the formation of the complex of bulge DNA and a ligand were estimated with basis set superposition error corrections incorporated into the elongation method.


Assuntos
DNA/química , Conformação de Ácido Nucleico , Química Computacional , Descoberta de Drogas , Ligantes , Teoria Quântica
3.
Nucleic Acids Res ; 49(16): 9479-9495, 2021 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-34358321

RESUMO

Fragile X-associated tremor/ataxia syndrome (FXTAS) is a late-onset neurodegenerative disorder caused by a limited expansion of CGG repeats in the FMR1 gene. Degeneration of neurons in FXTAS cell models can be triggered by accumulation of polyglycine protein (FMRpolyG), a by-product of translation initiated upstream to the repeats. Specific aims of our work included testing if naphthyridine-based molecules could (i) block FMRpolyG synthesis by binding to CGG repeats in RNA, (ii) reverse pathological alterations in affected cells and (iii) preserve the content of FMRP, translated from the same FMR1 mRNA. We demonstrate that cyclic mismatch binding ligand CMBL4c binds to RNA structure formed by CGG repeats and attenuates translation of FMRpolyG and formation of nuclear inclusions in cells transfected with vectors expressing RNA with expanded CGG repeats. Moreover, our results indicate that CMBL4c delivery can reduce FMRpolyG-mediated cytotoxicity and apoptosis. Importantly, its therapeutic potential is also observed once the inclusions are already formed. We also show that CMBL4c-driven FMRpolyG loss is accompanied by partial FMRP reduction. As complete loss of FMRP induces FXS in children, future experiments should aim at evaluation of CMBL4c therapeutic intervention in differentiated tissues, in which FMRpolyG translation inhibition might outweigh adverse effects related to FMRP depletion.

4.
Chem Commun (Camb) ; 57(26): 3235-3238, 2021 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-33646236

RESUMO

We demonstrated that a synthetic ligand NA, which selectively binds to a 5'-CAG-3'/5'-CAG-3' triad, induced repeat contractions during DNA polymerase-mediated primer extension through the CAG repeat template. A thorough capillary electrophoresis and sequencing analysis revealed that the d(CAG)20 template gave shortened nascent strands mainly containing 3-6 CTG units in the presence of NA.


Assuntos
DNA/genética , Naftiridinas/farmacologia , Quinolonas/farmacologia , Repetições de Trinucleotídeos/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , Eletroforese Capilar , Humanos , Ligantes , Conformação de Ácido Nucleico/efeitos dos fármacos
5.
Bioorg Med Chem ; 36: 116070, 2021 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-33773376

RESUMO

RNA is an emerging target of next-generation drug development. Recently, new small molecules targeting RNAs were discovered by several pharmaceutical companies. Methods have been reported to identify small molecules targeting a specific RNA sequence and structural motif, however, because of diverse sequence and structural motifs potentially present in the druggable functional RNAs, large sets of structure-activity relationships (SARs) information of small molecule - RNA interactions will be required for the acceleration and efficient startup of the discovery programs toward unprecedented RNA targets. Here we describe our iterative RNA selection and compounds screening to accumulate rich information about small molecules - RNA interaction. The RNAs that selectively bind to the initial molecular target, compound 1 from our in-house chemical library (JT-library), was isolated using in vitro selection technique from a hairpin-structured RNA library mimicking precursor microRNA (pre-miRNA). Then, we engineered pre-let-7f-2 to create its mutant that can bind to compound 1 by embedding the in vitro selected RNA motif for compound 1 in the hairpin loop region. The obtained mutant pre-let-7f-2-loop-mt was used as a target for screening 316 analogs of compound 1. A surface plasmon resonance (SPR) -based screening was performed against pre-let-7f-2-loop-mt-immobilized sensor surface and we obtained four compounds that can bind to the RNA. Among these four compounds, three compounds showed higher affinity to pre-let-7f-2-loop-mt than the parental compound 1, which suggests the feasibility of our strategy for gathering the SAR information on small molecule - RNA interactions. We demonstrated only one cycle of RNA selection and compounds screening in the present study, but can continue this cycle with the selected molecule to gain new RNAs and even new RNA motifs and gather much SAR information with improved accuracy.


Assuntos
Descoberta de Drogas , RNA/antagonistas & inibidores , Bibliotecas de Moléculas Pequenas/farmacologia , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Humanos , Estrutura Molecular , RNA/química , Bibliotecas de Moléculas Pequenas/química , Relação Estrutura-Atividade
6.
Biochemistry ; 60(4): 245-249, 2021 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-33476116

RESUMO

MicroRNAs are potential targets for drug development. Small molecules that can inhibit or promote a specific miRNA's biogenesis would be useful for regulating its target genes. Various types of small molecules have been investigated so far for their potential application in modulating miRNA biogenesis. They bind to the target primary or precursor miRNAs and inhibit the processing of these precursors by Drosha or Dicer. However, the binding site that effectively interferes with the Dicer cleavage reaction is still undetermined. Here we report that our designed small molecule restricted naphthyridine dimer (RND) binds to the hairpin loop of a hairpin RNA and induces its dimerization. This study shows that the binding of the RND to the hairpin loop was not effective in interfering with the Dicer cleavage reaction, but dimerization of the hairpin RNA by RND binding effectively interfered with the Dicer cleavage reaction.


Assuntos
RNA Helicases DEAD-box/química , MicroRNAs/química , Modelos Químicos , Conformação de Ácido Nucleico , Ribonuclease III/química , Humanos
7.
Nat Commun ; 12(1): 236, 2021 01 11.
Artigo em Inglês | MEDLINE | ID: mdl-33431896

RESUMO

Synthetic small molecules modulating RNA structure and function have therapeutic potential for RNA diseases. Here we report our discovery that naphthyridine carbamate dimer (NCD) targets disease-causing r(UGGAA)n repeat RNAs in spinocerebellar ataxia type 31 (SCA31). Structural analysis of the NCD-UGGAA/UGGAA complex by nuclear magnetic resonance (NMR) spectroscopy clarifies the mode of binding that recognizes four guanines in the UGGAA/UGGAA pentad by hydrogen bonding with four naphthyridine moieties of two NCD molecules. Biological studies show that NCD disrupts naturally occurring RNA foci built on r(UGGAA)n repeat RNA known as nuclear stress bodies (nSBs) by interfering with RNA-protein interactions resulting in the suppression of nSB-mediated splicing events. Feeding NCD to larvae of the Drosophila model of SCA31 alleviates the disease phenotype induced by toxic r(UGGAA)n repeat RNA. These studies demonstrate that small molecules targeting toxic repeat RNAs are a promising chemical tool for studies on repeat expansion diseases.


Assuntos
Drosophila/genética , RNA/genética , Animais , Sequência de Bases , Núcleo Celular/metabolismo , Modelos Animais de Doenças , Células HeLa , Humanos , Íntrons/genética , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Conformação de Ácido Nucleico , Fenótipo , Fosforilação , Proteínas de Ligação a RNA/metabolismo , Bibliotecas de Moléculas Pequenas/farmacologia , Temperatura
8.
Biochemistry ; 59(29): 2679-2683, 2020 07 28.
Artigo em Inglês | MEDLINE | ID: mdl-32628834

RESUMO

The methylation of cytosine in the full mutation of the expanded CGG repeat and subsequent deamination to thymine could be a measure of repeat instability. We report the synthesis of NCD-Bpy, which binds to the TGG/CGG site in the repeat hairpin. NCD-Bpy forces the thymine in the TGG/CGG site to flip out from the π-stack, recruits osmium tetroxide in the vicinity of the flipped-out T, and oxidizes the T. The piperidine-induced cleavage band successfully determined the position of the T in the expanded CGG repeat.


Assuntos
2,2'-Dipiridil/química , 5-Metilcitosina/análise , Naftiridinas/química , Timina/análise , Repetições de Trinucleotídeos , 2,2'-Dipiridil/síntese química , Desaminação , Metilação , Naftiridinas/síntese química , Expansão das Repetições de Trinucleotídeos
9.
Chemistry ; 26(63): 14305-14309, 2020 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-32449537

RESUMO

Expanded CUG repeat RNA in the dystrophia myotonia protein kinase (DMPK) gene causes myotonic dystrophy type 1 (DM1) and sequesters RNA processing proteins, such as the splicing factor muscleblind-like 1 protein (MBNL1). Sequestration of splicing factors results in the mis-splicing of some pre-mRNAs. Small molecules that rescue the mis-splicing in the DM1 cells have drawn attention as potential drugs to treat DM1. Herein we report a new molecule JM642 consisted of two 1,3-diaminoisoquinoline chromophores having an auxiliary aromatic unit at the C5 position. JM642 alternates the splicing pattern of the pre-mRNA of the Ldb3 gene in the DM1 cell model and Clcn1 and Atp2a1 genes in the DM1 mouse model. In vitro binding analysis by surface plasmon resonance (SPR) assay to the r(CUG) repeat and disruption of ribonuclear foci in the DM1 cell model suggested the binding of JM642 to the expanded r(CUG) repeat in vivo, eventually rescue the mis-splicing.


Assuntos
Distrofia Miotônica , Quinolinas , Splicing de RNA , Animais , Canais de Cloreto/genética , Canais de Cloreto/metabolismo , Dimerização , Camundongos , Distrofia Miotônica/tratamento farmacológico , Distrofia Miotônica/genética , Quinolinas/química , Quinolinas/farmacologia , RNA/metabolismo , Splicing de RNA/efeitos dos fármacos , Proteínas de Ligação a RNA/metabolismo , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/genética , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Expansão das Repetições de Trinucleotídeos
10.
Biochem Biophys Res Commun ; 531(1): 56-61, 2020 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-32278549

RESUMO

An expanded GGGGCC hexanucleotide (G4C2) repeat within the non-coding region of C9ORF72 gene has been identified as the most common genetic cause of FTD/ALS kindred, and synthetic ligand targeting this pathological expansion sequence holds a promising approach for the disease interference. We here describe the naphthyridine carbamate tetramer, p-NCTB, as a binding ligand to hairpin G4C2 repeat. p-NCTB simultaneously recognizes two distal CGGG/CGGG sites in G4C2 repeat DNA and RNA leading to the formation of the interhelical (inter- and intrastrand) binding complexes. The intrastrand binding was predominant when p-NCTB bound to long repeat sequence that accommodates multiple binding sites by folding into hairpins, while the interstrand binding was exclusive for short repeat sequence. The binding of p-NCTB showed repeat-length selectivity: the longer repeat sequence is a better target for p-NCTB. p-NCTB demonstrated inhibition of transcription against G4C2 repeat template in vitro in a repeat length-dependent manner.


Assuntos
Proteína C9orf72/genética , Expansão das Repetições de DNA/efeitos dos fármacos , Naftiridinas/farmacologia , Transcrição Genética/efeitos dos fármacos , Esclerose Amiotrófica Lateral/tratamento farmacológico , Esclerose Amiotrófica Lateral/genética , Demência Frontotemporal/tratamento farmacológico , Demência Frontotemporal/genética , Humanos , Naftiridinas/química , RNA/genética , Sequências Repetitivas de Ácido Nucleico/efeitos dos fármacos
11.
Chem Commun (Camb) ; 56(39): 5227-5230, 2020 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-32342975

RESUMO

Herein, we show two new DNA binding small molecules, NCD-RO and NCD-RC, and their ability to bind and selectively assemble ruthenium complexes on G-G mismatch DNA. This study used a naphthyridine carbamate dimer (NCD) as an efficient scaffold to assemble metal complexes in a controlled manner on dsDNA.


Assuntos
Carbamatos/química , Complexos de Coordenação/química , DNA/química , Naftiridinas/química , Rutênio/química , Pareamento Incorreto de Bases , Dimerização , Ligantes , Estrutura Molecular
12.
Nat Genet ; 52(2): 146-159, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32060489

RESUMO

In many repeat diseases, such as Huntington's disease (HD), ongoing repeat expansions in affected tissues contribute to disease onset, progression and severity. Inducing contractions of expanded repeats by exogenous agents is not yet possible. Traditional approaches would target proteins driving repeat mutations. Here we report a compound, naphthyridine-azaquinolone (NA), that specifically binds slipped-CAG DNA intermediates of expansion mutations, a previously unsuspected target. NA efficiently induces repeat contractions in HD patient cells as well as en masse contractions in medium spiny neurons of HD mouse striatum. Contractions are specific for the expanded allele, independently of DNA replication, require transcription across the coding CTG strand and arise by blocking repair of CAG slip-outs. NA-induced contractions depend on active expansions driven by MutSß. NA injections in HD mouse striatum reduce mutant HTT protein aggregates, a biomarker of HD pathogenesis and severity. Repeat-structure-specific DNA ligands are a novel avenue to contract expanded repeats.


Assuntos
Proteína Huntingtina/genética , Doença de Huntington/genética , Naftiridinas/farmacologia , Quinolonas/farmacologia , Expansão das Repetições de Trinucleotídeos/efeitos dos fármacos , Animais , Corpo Estriado/efeitos dos fármacos , DNA/metabolismo , Reparo de Erro de Pareamento de DNA/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , Modelos Animais de Doenças , Humanos , Proteína Huntingtina/metabolismo , Doença de Huntington/tratamento farmacológico , Doença de Huntington/patologia , Masculino , Camundongos , Camundongos Transgênicos , Instabilidade de Microssatélites , Mutação , Ribonucleases/metabolismo , Proteína de Ligação a TATA-Box/genética , Transcrição Genética
13.
Chembiochem ; 21(4): 477-480, 2020 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-31397042

RESUMO

MicroRNAs (miRNAs) are short RNAs that regulate the expression of complementary messenger RNAs and are involved in numerous human diseases. However, current detection techniques lack the sensitivity to detect miRNAs of low abundance. Moreover, at a length of 20-25 bases, miRNAs are too short for the reverse transcription (RT) polymerase chain reaction (PCR). Here we have developed a new, rapid, and simple miRNA detection system utilizing an RT primer containing a DNA tag at the 5'-end to increase the length of the cDNA. This strategy increases the length of the hybridized tagged primer and the complementary template DNA, as well as the melting temperature of the primer⋅template DNA duplex. PCR efficiency is thus increased, thereby enhancing miRNA detection sensitivity.


Assuntos
Primers do DNA/química , MicroRNAs/análise , Humanos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
14.
Chem Commun (Camb) ; 56(5): 754-757, 2020 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-31844850

RESUMO

We have designed and synthesized a novel naphthyridine tetramer, p-NCTB, for the recognition of tandem guanine-guanine (G-G) mismatches in DNA. p-NCTB possesses a p-biphenyl linker connecting two naphthyridine carbamate dimer (NCD) moieties that recognize G-G mismatches. p-NCTB preferentially bound to tandem G-G mismatches in dCGGG/dCGGG over dCGG/dCGG. Two dCGGG/dCGGG sites were simultaneously recognized and were noncovalently cross-linked via the formation of inter- and intrastrand complexes with p-NCTB. The intrastrand binding was more favorable, which could allow p-NCTB to bind selectively to a sequence containing multiple dCGGG/dCGGG sites.

15.
Nucleic Acids Res ; 47(20): 10906-10913, 2019 11 18.
Artigo em Inglês | MEDLINE | ID: mdl-31566242

RESUMO

The trinucleotide repeat expansion disorders (TREDs) constitute of a group of >40 hereditary neurodegenerative human diseases associated with abnormal expansion of repeated sequences, such as CAG repeats. The pathogenic factor is a transcribed RNA or protein whose function in the cell is compromised. The disorders are progressive and incurable. Consequently, many ongoing studies are oriented at developing therapies. We have analyzed crystal structures of RNA containing CAG repeats in complex with synthetic cyclic mismatch-binding ligands (CMBLs). The models show well-defined interactions between the molecules in which the CMBLs mimic nucleobases as they form pseudo-canonical base pairs with adenosine residues and engage in extensive stacking interactions with neighboring nucleotides. The binding of ligands is associated with major structural changes of the CAG repeats, which is consistent with results of biochemical studies. The results constitute an early characterization of the first lead compounds in the search for therapy against TREDs. The crystallographic data indicate how the compounds could be further refined in future biomedical studies.


Assuntos
RNA/genética , Expansão das Repetições de Trinucleotídeos/genética , Adenosina/metabolismo , Sequência de Bases , Ligantes , Conformação de Ácido Nucleico , Desnaturação de Ácido Nucleico , RNA/química , Solventes , Temperatura , Raios Ultravioleta
16.
Methods ; 167: 78-91, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31078794

RESUMO

Much recent attention has been focused on small organic molecules binding to non-canonical structures of nucleic acids, especially, RNA. The Human Genome Project and the ENCODE (encyclopedia of DNA elements) project revealed that more than 75% of the human genome is transcribed into RNA, while only ∼3% of the human genome encodes a protein. These non-protein-coding RNAs are thought to play significant roles in many cellular processes and are promising targets for drug discovery. Emerging roles of the non-coding RNAs in a variety of diseases provides enormous opportunities for pharmaceutical research on developing drugs targeting undruggable and rare diseases. During the last two decades, our laboratory has focused attention on small molecules binding to non-canonical DNA and RNA structures, especially to mismatched base pairs. Mismatch binding ligands (MBLs) we have developed are synthetic molecules designed in silico based on the hypothesis of hydrogen-bonding and semi-intercalation to DNA and RNA. Most of MBLs consists of two heterocycles having hydrogen bonding surfaces fully or partially complementary to that of nucleotide bases. In our design, each heterocycle binds to one of the mismatched bases by hydrogen bonding to form pseudo-base pairs, which would be stacked with the adjacent base pairs. The hypothesis allows us in principle to design small molecules binding to any mismatched base pairs, but it turned out not to be the case in reality. However, we have so far succeeded in developing several MBLs binding to DNA and RNA motifs of biological significance. In this review, we shall describe the hypothesis of molecular design of MBLs and its outcome regarding RNA targeting.


Assuntos
DNA/ultraestrutura , Conformação de Ácido Nucleico/efeitos dos fármacos , RNA/ultraestrutura , Bibliotecas de Moléculas Pequenas/química , Pareamento de Bases/genética , Simulação por Computador , DNA/efeitos dos fármacos , DNA/genética , Humanos , Ligação de Hidrogênio , Ligantes , Motivos de Nucleotídeos/genética , RNA/efeitos dos fármacos , RNA/genética , Bibliotecas de Moléculas Pequenas/farmacologia
17.
Chembiochem ; 20(23): 2903-2910, 2019 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-31087756

RESUMO

We introduce the concept of molecular glues for RNA, in which specific RNA-binding small molecules induce designed structural changes in target functional RNAs, resulting in modulation of the functions. (Z)-NCTS is an RNA-mismatch-binding small molecule that recognizes 5'-r(XGG)-3'/5'-r(XGG)-3' sequences (X=U or A) and acts as a molecular glue for RNA. The binding of (Z)-NCTS brings two distinct 5'-r(XGG)-3' domains into contact with each other, and this can result in higher-order structural changes of target RNAs. We applied (Z)-NCTS to induce the formation of a proposed tertiary structure of a ribozyme together with activation of RNA-cleaving ability. The concept of a molecular glue could inspire new small-molecule-based strategies for regulating biological functions: a synthetic small molecule targeting functional RNAs could regulate the RNA structure and function.


Assuntos
Naftiridinas/farmacologia , Conformação de Ácido Nucleico/efeitos dos fármacos , RNA Catalítico/metabolismo , DNA de Cadeia Simples/genética , DNA de Cadeia Simples/metabolismo , Expressão Gênica/efeitos dos fármacos , Células HeLa , Humanos , Sequências Repetidas Invertidas , Naftiridinas/metabolismo , Hibridização de Ácido Nucleico/efeitos dos fármacos , RNA Catalítico/genética
18.
Bioorg Med Chem ; 27(10): 2140-2148, 2019 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-30952388

RESUMO

Small-molecule modulators, along with antisense oligonucleotide, would be powerful tools and potential drug candidates for modulating miRNA-related gene expressions. The mechanism of the inhibitory effect of the C-bulge binding small molecule BzDANP for the Dicer processing reaction of pre-miR-136 was discussed on the data obtained by SPR, NMR, and kinetic analysis for Dicer processing. SPR and NMR analysis showed the preference of BzDANP binding to the C-bulge. Michaelis-Menten analysis suggested the formation of a ternary complex pre-miR-136-BzDANP-Dicer during the Dicer-cleavage reaction of pre-miR-136 in the presence of BzDANP. The inhibitory effect of BzDANP is likely attributed to the slower reaction from the ternary complex than that from the binary pre-miR-136-Dicer complex.


Assuntos
RNA Helicases DEAD-box/metabolismo , MicroRNAs/química , Naftiridinas/química , Ribonuclease III/metabolismo , Bibliotecas de Moléculas Pequenas/química , RNA Helicases DEAD-box/antagonistas & inibidores , Espectroscopia de Ressonância de Spin Eletrônica , Humanos , Espectroscopia de Ressonância Magnética , MicroRNAs/genética , MicroRNAs/metabolismo , Mutação , Naftiridinas/metabolismo , Conformação de Ácido Nucleico , Ligação Proteica , Precursores de RNA/química , Precursores de RNA/genética , Precursores de RNA/metabolismo , Ribonuclease III/antagonistas & inibidores , Bibliotecas de Moléculas Pequenas/metabolismo
19.
Chemistry ; 24(68): 18115-18122, 2018 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-30302858

RESUMO

Expanded r(CUG) repeats are the cause of the neurological disorder myotonic dystrophy type 1 (DM1). The pathological features of DM1 include the formation of ribonuclear foci containing expanded r(CUG) repeats, which sequester the MBNL1 protein and lead to the misregulation of alternative pre-mRNA splicing. Small molecules that bind to the r(CUG) repeats and improve alternative splicing have therapeutic potential in the treatment of DM1. Herein, the synthesis of DDAP (a dimeric form of the CUG-binding molecule DAP reported previously), its binding properties to r(CUG) repeats, and its effect on the misregulation of splicing are reported. The surface plasmon resonance assay, circular dichroism spectra, and ESI-TOF mass spectrometry results confirmed the binding of DDAP to r(CUG)9 repeats. Studies on a DM1 cell model and a DM1 mouse model revealed that DDAP was partially effective in the recovery of the pre-mRNA splicing defects. The mechanism underlying this recovery was studied in vitro through a competitive binding assay, and suggested that DDAP could interfere with the binding of MBNL1 to r(CUG) repeats in a concentration-dependent manner.


Assuntos
Processamento Alternativo/efeitos dos fármacos , Distrofia Miotônica/tratamento farmacológico , Distrofia Miotônica/genética , Fenantrolinas/química , Fenantrolinas/farmacologia , Animais , Proteínas de Ligação a DNA/genética , Modelos Animais de Doenças , Descoberta de Drogas , Humanos , Ligantes , Camundongos , RNA Mensageiro/genética , Proteínas de Ligação a RNA/genética
20.
Chem Commun (Camb) ; 54(93): 13072-13075, 2018 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-30280156

RESUMO

A new DNA binding small molecule, NCD-CC is reported. NCD-CC has a NCD domain, which recognizes the G-G mismatch in a CGG/CGG triad, and a cysteinylcystein (CC) moiety. Dimerization of NCD-CC through intermolecular disulfide bond formation was accelerated in the presence of CGG repeat DNA.


Assuntos
DNA/química , Dipeptídeos/química , Dissulfetos/síntese química , Naftiridinas/química , Sítios de Ligação , Dimerização , Dissulfetos/química , Humanos , Estrutura Molecular , Repetições de Trinucleotídeos
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