Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 25
Filtrar
1.
Chem Rec ; 21(4): 616-630, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33570227

RESUMO

This Personal Account describes the authors' foray into DNA-encoded libraries. The article addresses several key aspects of this hit generation technology, from the development of new synthetic methodology to the subsequent conception, design, and delivery of a DNA-encoded library. In particular, we have been engaged in adapting photocatalytic reactions to the idiosyncratic requirements of DNA-encoded chemistry. We have chosen one such methodology, namely a photocatalytic [2+2] cycloaddition reaction, to showcase how we employed property-based computational analyses to guide the selection and validation of building blocks for the production of a library. Ultimately, these novel bond disconnections and design principles led to the assembly of a DNA-encoded library that is composed of structurally diverse compounds within largely desirable property space and, therefore, well positioned to deliver novel chemical matter for drug discovery programs.


Assuntos
DNA/química , Desenho de Fármacos , Biblioteca Gênica , Bibliotecas de Moléculas Pequenas/síntese química , Catálise , Técnicas de Química Combinatória , Processos Fotoquímicos , Bibliotecas de Moléculas Pequenas/química
2.
Bioorg Med Chem ; 41: 116205, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-34000509

RESUMO

The ability to predict chemical structure from DNA sequence has to date been a necessary cornerstone of DNA-encoded library technology. DNA-encoded libraries (DELs) are typically screened by immobilized affinity selection and enriched library members are identified by counting the number of times an individual compound's sequence is observed in the resultant dataset. Those with high signal reads (DEL hits) are subsequently followed up through off-DNA synthesis of the predicted small molecule structures. However, hits followed-up in this manner often fail to translate to confirmed ligands. To address this low conversion rate of DEL hits to off-DNA ligands, we have developed an approach that eliminates the reliance on chemical structure prediction from DNA sequence. Here we describe our method of combining non-combinatorial resynthesis on-DNA following library procedures as a rapid means to assess the probable molecules attached to the DNA barcode. Furthermore, we apply our Bead-Assisted Ligand Isolation Mass Spectrometry (BALI-MS) technique to identify the true binders found within the mixtures of on-DNA synthesis products. Finally, we describe a Normalized Enrichment (NE) metric that allows for the quantitative assessment of affinity selection in these studies. We exemplify how this combined approach enables the identification of putative hit matter against a clinically relevant therapeutic target bisphosphoglycerate mutase, BPGM.


Assuntos
DNA/química , Descoberta de Drogas , Biblioteca Gênica , Espectrometria de Massas/métodos , Técnicas de Química Combinatória , Ligantes , Estrutura Molecular , Bibliotecas de Moléculas Pequenas/química
3.
Biochem Biophys Res Commun ; 533(2): 201-208, 2020 12 03.
Artigo em Inglês | MEDLINE | ID: mdl-32414574

RESUMO

A catalytic manifold that enables photoredox cross-electrophile coupling of alkyl bromides with DNA-tagged aryl iodides in aqueous solution is presented. This metallaphotoredox transformation was aided by the identification of a new pyridyl bis(carboxamidine) ligand, which proved critical to the nickel catalytic cycle. The described C(sp2)-C(sp3) coupling tolerates a wide range of both DNA-tagged aryl iodides as well as alkyl bromides. Importantly, this reaction was optimized for parallel synthesis, which is a paramount prerequisite for the preparation of combinatorial libraries, by using a 96-well plate-compatible blue LED array as the light source. Therefore, this mild and DNA-compatible transformation is well positioned for the construction of DNA-encoded libraries.


Assuntos
Alcanos/química , Brometos/química , DNA/química , Hidrocarbonetos Aromáticos/química , Iodetos/química , Alcanos/síntese química , Alquilação , Brometos/síntese química , Catálise , Técnicas de Química Combinatória , DNA/síntese química , Hidrocarbonetos Aromáticos/síntese química , Iodetos/síntese química , Ligantes , Níquel/química , Oxirredução , Bibliotecas de Moléculas Pequenas/síntese química , Bibliotecas de Moléculas Pequenas/química
4.
Angew Chem Int Ed Engl ; 59(19): 7377-7383, 2020 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-32050046

RESUMO

DNA encoded libraries (DEL) have shown promise as a valuable technology for democratizing the hit discovery process. Although DEL provides relatively inexpensive access to libraries of unprecedented size, their production has been hampered by the idiosyncratic needs of the encoding DNA tag relegating DEL compatible chemistry to dilute aqueous environments. Recently reversible adsorption to solid support (RASS) has been demonstrated as a promising method to expand DEL reactivity using standard organic synthesis protocols. Here we demonstrate a suite of on-DNA chemistries to incorporate medicinally relevant and C-S, C-P and N-S linkages into DELs, which are underrepresented in the canonical methods.


Assuntos
DNA/síntese química , Adsorção , Técnicas de Química Sintética , Técnicas de Química Combinatória , Descoberta de Drogas , Indicadores e Reagentes , Bibliotecas de Moléculas Pequenas , Solubilidade , Sulfonas/química , Sulfóxidos/química
5.
J Am Chem Soc ; 141(25): 9998-10006, 2019 06 26.
Artigo em Inglês | MEDLINE | ID: mdl-31136164

RESUMO

DNA Encoded Libraries have proven immensely powerful tools for lead identification. The ability to screen billions of compounds at once has spurred increasing interest in DEL development and utilization. Although DEL provides access to libraries of unprecedented size and diversity, the idiosyncratic and hydrophilic nature of the DNA tag severely limits the scope of applicable chemistries. It is known that biomacromolecules can be reversibly, noncovalently adsorbed and eluted from solid supports, and this phenomenon has been utilized to perform synthetic modification of biomolecules in a strategy we have described as reversible adsorption to solid support (RASS). Herein, we present the adaptation of RASS for a DEL setting, which allows reactions to be performed in organic solvents at near anhydrous conditions opening previously inaccessible chemical reactivities to DEL. The RASS approach enabled the rapid development of C(sp2)-C(sp3) decarboxylative cross-couplings with broad substrate scope, an electrochemical amination (the first electrochemical synthetic transformation performed in a DEL context), and improved reductive amination conditions. The utility of these reactions was demonstrated through a DEL-rehearsal in which all newly developed chemistries were orchestrated to afford a compound rich in diverse skeletal linkages. We believe that RASS will offer expedient access to new DEL reactivities, expanded chemical space, and ultimately more drug-like libraries.


Assuntos
Compostos de Anilina/síntese química , Técnicas de Química Combinatória/métodos , DNA/química , Piperidinas/síntese química , Compostos de Amônio Quaternário/química , Estudo de Prova de Conceito
6.
J Chem Inf Model ; 59(11): 4645-4653, 2019 11 25.
Artigo em Inglês | MEDLINE | ID: mdl-31689098

RESUMO

DNA encoded libraries (DEL) are being used as a complement or alternative to traditional high throughput screening (HTS). To maximize the chances of finding chemically attractive lead material that is appropriate for medicinal chemistry optimization, for example, in the Rule of Five compliant chemistry space, it is important to design DEL library compounds such that they are highly diverse and fall within a desired property space. Currently available library design methods can be classified as either monomer-based or product-based. As monomers may undergo significant structural changes when participating in a reaction, monomer based design can provide a poor representation of the properties of resultant DEL products. However, product-based design introduces a technical obstacle due to the enormous chemical design space for many DELs. Here a new method for monomer based selections is described using representative sublibraries as surrogates for fully enumerated DEL property-based optimization. Through a series of rational and systematic library enumerations and property calculations, building-block representatives are identified and representative sublibraries are defined to drive the optimization process. A published data set for a triazine library was used to demonstrate the effectiveness of the multiple objective optimization for six properties. All of the evaluated properties for the designed library are shown to consistently shift toward the desired property distribution as driven by the design criteria.


Assuntos
DNA/química , Bibliotecas de Moléculas Pequenas/química , Química Farmacêutica , Técnicas de Química Combinatória , DNA/síntese química , Descoberta de Drogas , Biblioteca Gênica , Humanos , Relação Quantitativa Estrutura-Atividade , Bibliotecas de Moléculas Pequenas/síntese química
7.
Bioorg Med Chem Lett ; 22(18): 5989-94, 2012 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-22892121
8.
SLAS Discov ; 26(2): 263-280, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33412987

RESUMO

Over the past 20 years, the toolbox for discovering small-molecule therapeutic starting points has expanded considerably. Pharmaceutical researchers can now choose from technologies that, in addition to traditional high-throughput knowledge-based and diversity screening, now include the screening of fragment and fragment-like libraries, affinity selection mass spectrometry, and selection against DNA-encoded libraries (DELs). Each of these techniques has its own unique combination of advantages and limitations that makes them more, or less, suitable for different target classes or discovery objectives, such as desired mechanism of action. Layered on top of this are the constraints of the drug-hunters themselves, including budgets, timelines, and available platform capacity; each of these can play a part in dictating the hit identification strategy for a discovery program. In this article, we discuss some of the factors that we use to govern our building of a hit identification roadmap for a program and describe the increasing role that DELs are playing in our discovery strategy. Furthermore, we share our learning during our initial exploration of DEL and highlight the approaches we have evolved to maximize the value returned from DEL selections. Topics addressed include the optimization of library design and production, reagent validation, data analysis, and hit confirmation. We describe how our thinking in these areas has led us to build a DEL platform that has begun to deliver tractable matter to our global discovery portfolio.


Assuntos
Descoberta de Drogas/métodos , Biblioteca Gênica , Bibliotecas de Moléculas Pequenas , Descoberta de Drogas/normas , Humanos
9.
Org Lett ; 22(8): 2908-2913, 2020 04 17.
Artigo em Inglês | MEDLINE | ID: mdl-32239950

RESUMO

The on-DNA synthesis of highly substituted cyclobutanes was achieved through a photocatalytic [2 + 2] cycloaddition reaction in aqueous solution. Readily available DNA-tagged styrene derivatives were reacted with structurally diverse cinnamates in the presence of an iridium-based photocatalyst, Ir(ppy)2(dtbbpy)PF6, to forge two new C(sp3)-C(sp3) bonds. This transformation was demonstrated to have excellent functional group tolerance and allowed for the facile installation of a variety of heteroaromatic substituents on a densely functionalized cyclobutane scaffold.


Assuntos
Ciclobutanos/química , DNA/síntese química , Catálise , Reação de Cicloadição , DNA/química , Estrutura Molecular , Processos Fotoquímicos
10.
ACS Med Chem Lett ; 11(6): 1101-1110, 2020 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-32550988

RESUMO

High-throughput experimentation (HTE) has emerged as an important tool in drug discovery, providing a platform for preparing large compound libraries and enabling swift reaction screening over wide-ranging conditions. Recent advances in automated high-density, material-sparing HTE have necessitated the development of rapid analytics with sensitivity and resolution sufficient to identify products and/or assess reaction performance in a timely and data-rich manner. Combination of an ultrathroughput (UT) reader platform with Acoustic Droplet Ejection-Open Port Interface-Mass Spectrometry (ADE-OPI-MS) provides the requisite speed and sensitivity. Herein, we report the application of ADE-OPI-MS to HTE in the areas of parallel medicinal chemistry and reaction screening.

11.
Chem Sci ; 11(45): 12282-12288, 2020 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-34094436

RESUMO

DNA-encoded library (DEL) technology has the potential to dramatically expedite hit identification in drug discovery owing to its ability to perform protein affinity selection with millions or billions of molecules in a few experiments. To expand the molecular diversity of DEL, it is critical to develop different types of DNA-encoded transformations that produce billions of molecules with distinct molecular scaffolds. Sequential functionalization of multiple C-H bonds provides a unique avenue for creating diversity and complexity from simple starting materials. However, the use of water as solvent, the presence of DNA, and the extremely low concentration of DNA-encoded coupling partners (0.001 M) have hampered the development of DNA-encoded C(sp3)-H activation reactions. Herein, we report the realization of palladium-catalyzed C(sp3)-H arylation of aliphatic carboxylic acids, amides and ketones with DNA-encoded aryl iodides in water. Notably, the present method enables the use of alternative sets of monofunctional building blocks, providing a linchpin to facilitate further setup for DELs. Furthermore, the C-H arylation chemistry enabled the on-DNA synthesis of structurally-diverse scaffolds containing enriched C(sp3) character, chiral centers, cyclopropane, cyclobutane, and heterocycles.

12.
ACS Comb Sci ; 21(8): 588-597, 2019 08 12.
Artigo em Inglês | MEDLINE | ID: mdl-31283168

RESUMO

A new catalytic manifold that merges photoredox with nickel catalysis in aqueous solution is presented. Specifically, the combination of a highly active, yet air-stable, nickel precatalyst with a new electron-deficient pyridyl carboxamidine ligand was key to the development of a water-compatible nickel catalysis platform, which is a crucial requirement for the preparation of DNA-encoded libraries (DELs). Together with an iridium-based photocatalyst and a powerful light source, this dual catalysis approach enabled the efficient decarboxylative arylation of α-amino acids with DNA-tagged aryl halides. This C(sp2)-C(sp3) coupling tolerates a wide variety of functional groups on both the amino acid and the aryl halide substrates. Due to the mild and DNA-compatible reaction conditions, the presented transformation holds great potential for the construction of DELs. This was further evidenced by showing that well plate-compatible LED arrays can serve as competent light sources to facilitate parallel synthesis. Lastly, we demonstrate that this procedure can serve as a blueprint toward the adaptation of other established nickel metallaphotoredox transformations to the idiosyncratic requirements of a DEL.


Assuntos
DNA/química , Níquel/química , Água/química , Catálise , Técnicas de Química Combinatória , Descarboxilação , Ligantes , Estrutura Molecular , Oxirredução , Processos Fotoquímicos , Soluções
13.
ACS Comb Sci ; 21(10): 650-655, 2019 10 14.
Artigo em Inglês | MEDLINE | ID: mdl-31425646

RESUMO

DNA-encoded chemical library (DECL) synthesis must occur in aqueous media under conditions that preserve the integrity of the DNA encoding tag. While the identification of "DNA-compatible" reaction conditions is critical for the development of DECL designs that explore previously inaccessible chemical space, reports measuring such compatibility have been largely restricted to methods that do not faithfully capture the impact of reaction conditions on DNA fidelity in solution phase. Here we report a comprehensive methodology that uses soluble DNA substrates that exactly recapitulate DNA's exposure to the chemically reactive species of DECL synthesis. This approach includes the assessment of chemical fidelity (reaction yield and purity), encoding fidelity (ligation efficiency), and readability (DNA compatibility), revealing the fate of the DNA tag during DECL chemistry from a single platform.


Assuntos
DNA/química , Bibliotecas de Moléculas Pequenas/síntese química , Técnicas de Química Combinatória , Estrutura Molecular , Bibliotecas de Moléculas Pequenas/química , Soluções
14.
ChemMedChem ; 13(20): 2159-2165, 2018 10 22.
Artigo em Inglês | MEDLINE | ID: mdl-30063289

RESUMO

A new procedure for the photoredox-mediated conjugate addition of radicals that can be conveniently generated from α-amino acids to DNA-tagged Michael acceptors and styrenes is presented. This C(sp3 )-C(sp3 ) coupling tolerates a broad array of structurally diverse radical precursors, including all of the 20 proteinogenic amino acids. Importantly, this reaction proceeds under mild conditions and in DNA-compatible aqueous media. Furthermore, the presented reaction conditions are compatible with DNA, making this reaction platform well suited for the construction of DNA-encoded libraries. The scope and limitations of the chemistry are discussed herein along with proposals for how this methodology might be used to construct DNA-encoded libraries.


Assuntos
Aminoácidos/química , DNA/química , Acrilamidas/química , Alquilação , Aminas/síntese química , Aminoácidos/efeitos da radiação , Catálise , Complexos de Coordenação/química , Complexos de Coordenação/efeitos da radiação , Descarboxilação , Radicais Livres/química , Irídio/química , Luz , Oxirredução , Estudo de Prova de Conceito
15.
J Med Chem ; 61(3): 1130-1152, 2018 02 08.
Artigo em Inglês | MEDLINE | ID: mdl-29298069

RESUMO

Janus kinases (JAKs) are intracellular tyrosine kinases that mediate the signaling of numerous cytokines and growth factors involved in the regulation of immunity, inflammation, and hematopoiesis. As JAK1 pairs with JAK2, JAK3, and TYK2, a JAK1-selective inhibitor would be expected to inhibit many cytokines involved in inflammation and immune function while avoiding inhibition of the JAK2 homodimer regulating erythropoietin and thrombopoietin signaling. Our efforts began with tofacitinib, an oral JAK inhibitor approved for the treatment of rheumatoid arthritis. Through modification of the 3-aminopiperidine linker in tofacitinib, we discovered highly selective JAK1 inhibitors with nanomolar potency in a human whole blood assay. Improvements in JAK1 potency and selectivity were achieved via structural modifications suggested by X-ray crystallographic analysis. After demonstrating efficacy in a rat adjuvant-induced arthritis (rAIA) model, PF-04965842 (25) was nominated as a clinical candidate for the treatment of JAK1-mediated autoimmune diseases.


Assuntos
Doenças Autoimunes/tratamento farmacológico , Ciclobutanos/farmacologia , Janus Quinase 1/antagonistas & inibidores , Inibidores de Proteínas Quinases/farmacologia , Pirimidinas/farmacologia , Pirróis/farmacologia , Sulfonamidas/farmacologia , Animais , Artrite Experimental/tratamento farmacológico , Ciclobutanos/química , Ciclobutanos/farmacocinética , Ciclobutanos/uso terapêutico , Cães , Avaliação Pré-Clínica de Medicamentos , Humanos , Concentração Inibidora 50 , Janus Quinase 1/química , Janus Quinase 2/antagonistas & inibidores , Modelos Moleculares , Conformação Proteica , Inibidores de Proteínas Quinases/química , Inibidores de Proteínas Quinases/farmacocinética , Inibidores de Proteínas Quinases/uso terapêutico , Pirimidinas/química , Pirimidinas/farmacocinética , Pirimidinas/uso terapêutico , Pirróis/química , Pirróis/farmacocinética , Pirróis/uso terapêutico , Ratos , Especificidade por Substrato , Sulfonamidas/química , Sulfonamidas/farmacocinética , Sulfonamidas/uso terapêutico , Distribuição Tecidual
16.
Sci Rep ; 6: 38573, 2016 12 09.
Artigo em Inglês | MEDLINE | ID: mdl-27934919

RESUMO

Inducing α-helicity through side-chain cross-linking is a strategy that has been pursued to improve peptide conformational rigidity and bio-availability. Here we describe the preparation of small peptides tethered to chiral sulfoxide-containing macrocyclic rings. Furthermore, a study of structure-activity relationships (SARs) disclosed properties with respect to ring size, sulfur position, oxidation state, and stereochemistry that show a propensity to induce α-helicity. Supporting data include circular dichroism spectroscopy (CD), NMR spectroscopy, and a single crystal X-ray structure for one such stabilized peptide. Finally, theoretical studies are presented to elucidate the effect of chiral sulfoxides in inducing backbone α-helicity.


Assuntos
Peptídeos/química , Conformação Proteica em alfa-Hélice , Safrol/análogos & derivados , Dicroísmo Circular , Modelos Moleculares , Oxirredução , Safrol/química
17.
J Med Chem ; 57(12): 5023-38, 2014 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-24417533

RESUMO

The Janus kinases (JAKs) are a family of intracellular tyrosine kinases that play an essential role in the signaling of numerous cytokines that have been implicated in the pathogenesis of inflammatory diseases. As a consequence, the JAKs have received significant attention in recent years from the pharmaceutical and biotechnology industries as therapeutic targets. Here, we provide a review of the JAK pathways, the structure, function, and activation of the JAK enzymes followed by a detailed look at the JAK inhibitors currently in the clinic or approved for these indications. Finally, a perspective is provided on what the past decade of research with JAK inhibitors for inflammatory indications has taught along with thoughts on what the future may hold in terms of addressing the opportunities and challenges that remain.


Assuntos
Anti-Inflamatórios/química , Doenças Autoimunes/tratamento farmacológico , Inflamação/tratamento farmacológico , Janus Quinases/antagonistas & inibidores , Inibidores de Proteínas Quinases/química , Animais , Anti-Inflamatórios/farmacologia , Antirreumáticos/farmacologia , Antirreumáticos/uso terapêutico , Doenças Autoimunes/imunologia , Doenças Autoimunes/metabolismo , Ensaios Clínicos como Assunto , Citocinas/metabolismo , Humanos , Inflamação/imunologia , Inflamação/metabolismo , Doenças Inflamatórias Intestinais/tratamento farmacológico , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/metabolismo , Janus Quinases/química , Janus Quinases/metabolismo , Piperidinas/farmacologia , Piperidinas/uso terapêutico , Conformação Proteica , Inibidores de Proteínas Quinases/farmacologia , Pirimidinas/farmacologia , Pirimidinas/uso terapêutico , Pirróis/farmacologia , Pirróis/uso terapêutico , Transdução de Sinais
18.
J Med Chem ; 57(23): 10072-9, 2014 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-25375838

RESUMO

Interest in drugs that covalently modify their target is driven by the desire for enhanced efficacy that can result from the silencing of enzymatic activity until protein resynthesis can occur, along with the potential for increased selectivity by targeting uniquely positioned nucleophilic residues in the protein. However, covalent approaches carry additional risk for toxicities or hypersensitivity reactions that can result from covalent modification of unintended targets. Here we describe methods for measuring the reactivity of covalent reactive groups (CRGs) with a biologically relevant nucleophile, glutathione (GSH), along with kinetic data for a broad array of electrophiles. We also describe a computational method for predicting electrophilic reactivity, which taken together can be applied to the prospective design of thiol-reactive covalent inhibitors.


Assuntos
Inibidores Enzimáticos/química , Glutationa/química , Desenho de Fármacos , Glutationa/metabolismo , Humanos , Cinética , Espectrometria de Massas , Ressonância Magnética Nuclear Biomolecular , Preparações Farmacêuticas/química
19.
J Med Chem ; 56(12): 5079-93, 2013 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-23735048

RESUMO

We report novel polymyxin analogues with improved antibacterial in vitro potency against polymyxin resistant recent clinical isolates of Acinetobacter baumannii and Pseudomonas aeruginosa . In addition, a human renal cell in vitro assay (hRPTEC) was used to inform structure-toxicity relationships and further differentiate analogues. Replacement of the Dab-3 residue with a Dap-3 in combination with a relatively polar 6-oxo-1-phenyl-1,6-dihydropyridine-3-carbonyl side chain as a fatty acyl replacement yielded analogue 5x, which demonstrated an improved in vitro antimicrobial and renal cytotoxicity profiles relative to polymyxin B (PMB). However, in vivo PK/PD comparison of 5x and PMB in a murine neutropenic thigh model against P. aeruginosa strains with matched MICs showed that 5x was inferior to PMB in vivo, suggesting a lack of improved therapeutic index in spite of apparent in vitro advantages.


Assuntos
Infecção Hospitalar/tratamento farmacológico , Descoberta de Drogas , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Polimixinas/química , Polimixinas/farmacologia , beta-Alanina/análogos & derivados , Animais , Antibacterianos/química , Antibacterianos/farmacocinética , Antibacterianos/farmacologia , Antibacterianos/toxicidade , Cães , Feminino , Bactérias Gram-Negativas/fisiologia , Humanos , Masculino , Testes de Sensibilidade Microbiana , Polimixinas/farmacocinética , Polimixinas/toxicidade , Ratos , beta-Alanina/química
20.
PLoS One ; 7(12): e51732, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23240059

RESUMO

Antibacterial compounds that affect bacterial viability have traditionally been identified, confirmed, and characterized in standard laboratory media. The historical success of identifying new antibiotics via this route has justifiably established a traditional means of screening for new antimicrobials. The emergence of multi-drug-resistant (MDR) bacterial pathogens has expedited the need for new antibiotics, though many in the industry have questioned the source(s) of these new compounds. As many pharmaceutical companies' chemical libraries have been exhaustively screened via the traditional route, we have concluded that all compounds with any antibacterial potential have been identified. While new compound libraries and platforms are being pursued, it also seems prudent to screen the libraries we currently have in hand using alternative screening approaches. One strategy involves screening under conditions that better reflect the environment pathogens experience during an infection, and identifying in vivo essential targets and pathways that are dispensable for growth in standard laboratory media in vitro. Here we describe a novel screening strategy for identifying compounds that inhibit the glyoxylate shunt in Pseudomonas aeruginosa, a pathway that is required for bacterial survival in the pulmonary environment. We demonstrate that these compounds, which were not previously identified using traditional screening approaches, have broad-spectrum antibacterial activity when they are tested under in vivo-relevant conditions. We also show that these compounds have potent activity on both enzymes that comprise the glyoxylate shunt, a feature that was supported by computational homology modeling. By dual-targeting both enzymes in this pathway, we would expect to see a reduced propensity for resistance development to these compounds. Taken together, these data suggest that understanding the in vivo environment that bacterial pathogens must tolerate, and adjusting the antibacterial screening paradigm to reflect those conditions, could identify novel antibiotics for the treatment of serious MDR pathogens.


Assuntos
Antibacterianos , Glioxilatos/metabolismo , Isocitrato Liase/antagonistas & inibidores , Malato Sintase/antagonistas & inibidores , Pseudomonas aeruginosa , Antibacterianos/química , Antibacterianos/uso terapêutico , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Glioxilatos/antagonistas & inibidores , Ensaios de Triagem em Larga Escala , Humanos , Isocitrato Liase/metabolismo , Malato Sintase/metabolismo , Redes e Vias Metabólicas/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/patogenicidade , Deleção de Sequência , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/farmacologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA