Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 13 de 13
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
J Pers Med ; 5(4): 354-69, 2015 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-26529019

RESUMO

"Cancer 2015" is a longitudinal and prospective cohort. It is a phased study whose aim was to pilot recruiting 1000 patients during phase 1 to establish the feasibility of providing a population-based genomics cohort. Newly diagnosed adult patients with solid cancers, with residual tumour material for molecular genomics testing, were recruited into the cohort for the collection of a dataset containing clinical, molecular pathology, health resource use and outcomes data. 1685 patients have been recruited over almost 3 years from five hospitals. Thirty-two percent are aged between 61-70 years old, with a median age of 63 years. Diagnostic tumour samples were obtained for 90% of these patients for multiple parallel sequencing. Patients identified with somatic mutations of potentially "actionable" variants represented almost 10% of those tumours sequenced, while 42% of the cohort had no mutations identified. These genomic data were annotated with information such as cancer site, stage, morphology, treatment and patient outcomes and health resource use and cost. This cohort has delivered its main objective of establishing an upscalable genomics cohort within a clinical setting and in phase 2 aims to develop a protocol for how genomics testing can be used in real-time clinical decision-making, providing evidence on the value of precision medicine to clinical practice.

2.
Drug Discov Today ; 20(12): 1429-32, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26494144

RESUMO

Genomic cancer medicine promises revolutionary change in oncology. The impacts of 'personalized medicine', based upon a molecular classification of cancer and linked to targeted therapies, will extend from individual patient outcomes to the health economy at large. To address the 'whole-of-system' impact of genomic cancer medicine, we have established a prospective cohort of patients with newly diagnosed cancer in the state of Victoria, Australia, about whom we have collected a broad range of clinical, demographic, molecular, and patient-reported data, as well as data on health resource utilization. Our goal is to create a model for investigating public investment in genomic medicine that maximizes the cost:benefit ratio for the Australian community at large.


Assuntos
Neoplasias/tratamento farmacológico , Neoplasias/genética , Austrália , Genômica/métodos , Humanos , Estudos Longitudinais , Medicina/métodos , Medicina de Precisão/métodos , Estudos Prospectivos
3.
PLoS One ; 10(10): e0139036, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26447468

RESUMO

Frameshifting during translation of viral or in rare cases cellular mRNA results in the synthesis of proteins from two overlapping reading frames within the same mRNA. In HIV-1 the protease, reverse transcriptase, and integrase enzymes are in a second reading frame relative to the structural group-specific antigen (gag), and their synthesis is dependent upon frameshifting. This ensures that a strictly regulated ratio of structural proteins and enzymes, which is critical for HIV-1 replication and viral infectivity, is maintained during protein synthesis. The frameshift element in HIV-1 RNA is an attractive target for the development of a new class of anti HIV-1 drugs. However, a number of examples are now emerging of human genes using -1 frameshifting, such as PEG10 and CCR5. In this study we have compared the HIV-1 and PEG10 frameshift elements and shown they have distinct functional characteristics. Frameshifting occurs at several points within each element. Moreover, frameshift modulators that were isolated by high-throughput screening of a library of 114,000 lead-like compounds behaved differently with the PEG10 frameshift element. The most effective compounds affecting the HIV-1 element enhanced frameshifting by 2.5-fold at 10 µM in two different frameshift reporter assay systems. HIV-1 protease:gag protein ratio was affected by a similar amount in a specific assay of virally-infected cultured cell, but the modulation of frameshifting of the first-iteration compounds was not sufficient to show significant effects on viral infectivity. Importantly, two compounds did not affect frameshifting with the human PEG10 element, while one modestly inhibited rather than enhanced frameshifting at the human element. These studies indicate that frameshift elements have unique characteristics that may allow targeting of HIV-1 and of other viruses specifically for development of antiviral therapeutic molecules without effect on human genes like PEG10 that use the same generic mechanism.


Assuntos
HIV-1/genética , Proteínas/metabolismo , RNA Viral/metabolismo , Bibliotecas de Moléculas Pequenas/química , Proteínas Reguladoras de Apoptose , Sequência de Bases , Cromatografia Líquida de Alta Pressão , Proteínas de Ligação a DNA , Mutação da Fase de Leitura , Genes Reporter , Células HEK293 , HIV-1/metabolismo , Humanos , Espectrometria de Massas , Conformação de Ácido Nucleico , Biossíntese de Proteínas , Proteínas de Ligação a RNA
4.
BMC Med Genomics ; 7: 23, 2014 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-24885028

RESUMO

BACKGROUND: Clinical specimens undergoing diagnostic molecular pathology testing are fixed in formalin due to the necessity for detailed morphological assessment. However, formalin fixation can cause major issues with molecular testing, as it causes DNA damage such as fragmentation and non-reproducible sequencing artefacts after PCR amplification. In the context of massively parallel sequencing (MPS), distinguishing true low frequency variants from sequencing artefacts remains challenging. The prevalence of formalin-induced DNA damage and its impact on molecular testing and clinical genomics remains poorly understood. METHODS: The Cancer 2015 study is a population-based cancer cohort used to assess the feasibility of mutational screening using MPS in cancer patients from Victoria, Australia. While blocks were formalin-fixed and paraffin-embedded in different anatomical pathology laboratories, they were centrally extracted for DNA utilising the same protocol, and run through the same MPS platform (Illumina TruSeq Amplicon Cancer Panel). The sequencing artefacts in the 1-10% and the 10-25% allele frequency ranges were assessed in 488 formalin-fixed tumours from the pilot phase of the Cancer 2015 cohort. All blocks were less than 2.5 years of age (mean 93 days). RESULTS: Consistent with the signature of DNA damage due to formalin fixation, many formalin-fixed samples displayed disproportionate levels of C>T/G>A changes in the 1-10% allele frequency range. Artefacts were less apparent in the 10-25% allele frequency range. Significantly, changes were inversely correlated with coverage indicating high levels of sequencing artefacts were associated with samples with low amounts of available amplifiable template due to fragmentation. The degree of fragmentation and sequencing artefacts differed between blocks sourced from different anatomical pathology laboratories. In a limited validation of potentially actionable low frequency mutations, a NRAS G12D mutation in a melanoma was shown to be a false positive. CONCLUSIONS: These findings indicate that DNA damage following formalin fixation remains a major challenge in laboratories working with MPS. Methodologies that assess, minimise or remove formalin-induced DNA damaged templates as part of MPS protocols will aid in the interpretation of genomic results leading to better patient outcomes.


Assuntos
Artefatos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Mutação/genética , Neoplasias/genética , Fixação de Tecidos , Linhagem Celular Tumoral , Fragmentação do DNA , Reações Falso-Positivas , Formaldeído , Humanos , Inclusão em Parafina , Reação em Cadeia da Polimerase , Estudos Prospectivos , Análise de Sequência de DNA , Moldes Genéticos , Uracila-DNA Glicosidase/metabolismo
5.
J Med Chem ; 57(4): 1323-43, 2014 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-24456288

RESUMO

The prosurvival BCL-2 proteins are attractive yet challenging targets for medicinal chemists. Their involvement in the initiation and progression of many, if not all, tumors makes them prime targets for developing new anticancer therapies. We present our approach based on de novo structure-based drug design. Using known structural information from complexes engaging opposing members of the BCL-2 family of proteins, we designed peptidomimetic compounds using a benzoylurea scaffold to reproduce key interactions between these proteins. A library stemming from the initial de novo designed scaffold led to the discovery of ligands with low micromolar potency (KD = 4 µM) and selectivity for BCL-XL. These compounds bind in the canonical BH3 binding groove in a binding mode distinct from previously known BCL-2 inhibitors. The results of our study provide insight into the design of a new class of antagonists targeting a challenging class of protein-protein interactions.


Assuntos
Ureia/análogos & derivados , Proteína bcl-X/antagonistas & inibidores , Cristalografia por Raios X , Desenho de Fármacos , Espectroscopia de Ressonância Magnética , Mimetismo Molecular , Estrutura Molecular , Espectrometria de Massas por Ionização por Electrospray , Relação Estrutura-Atividade , Ureia/síntese química , Ureia/farmacologia , Proteína bcl-X/química
6.
J Med Chem ; 56(13): 5514-40, 2013 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-23767404

RESUMO

Developing potent molecules that inhibit Bcl-2 family mediated apoptosis affords opportunities to treat cancers via reactivation of the cell death machinery. We describe the hit-to-lead development of selective Bcl-XL inhibitors originating from a high-throughput screening campaign. Small structural changes to the hit compound increased binding affinity more than 300-fold (to IC50 < 20 nM). This molecular series exhibits drug-like characteristics, low molecular weights (Mw < 450), and unprecedented selectivity for Bcl-XL. Surface plasmon resonance experiments afford strong evidence of binding affinity within the hydrophobic groove of Bcl-XL. Biological experiments using engineered Mcl-1 deficient mouse embryonic fibroblasts (MEFs, reliant only on Bcl-XL for survival) and Bax/Bak deficient MEFs (insensitive to selective activation of Bcl-2-driven apoptosis) support a mechanism-based induction of apoptosis. This manuscript describes the first series of selective small-molecule inhibitors of Bcl-XL and provides promising leads for the development of efficacious therapeutics against solid tumors and chemoresistant cancer cell lines.


Assuntos
Apoptose/efeitos dos fármacos , Benzotiazóis/farmacologia , Hidrazonas/farmacologia , Proteína bcl-X/antagonistas & inibidores , Animais , Benzotiazóis/síntese química , Benzotiazóis/metabolismo , Ligação Competitiva , Linhagem Celular Tumoral , Células Cultivadas , Descoberta de Drogas , Embrião de Mamíferos/citologia , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Hidrazonas/síntese química , Hidrazonas/metabolismo , Cinética , Camundongos , Camundongos Knockout , Modelos Químicos , Estrutura Molecular , Proteína de Sequência 1 de Leucemia de Células Mieloides/deficiência , Proteína de Sequência 1 de Leucemia de Células Mieloides/genética , Ressonância de Plasmônio de Superfície , Proteína Killer-Antagonista Homóloga a bcl-2/deficiência , Proteína Killer-Antagonista Homóloga a bcl-2/genética , Proteína X Associada a bcl-2/deficiência , Proteína X Associada a bcl-2/genética , Proteína bcl-X/química , Proteína bcl-X/metabolismo
7.
Nat Chem Biol ; 9(6): 390-7, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23603658

RESUMO

The prosurvival BCL-2 family protein BCL-X(L) is often overexpressed in solid tumors and renders malignant tumor cells resistant to anticancer therapeutics. Enhancing apoptotic responses by inhibiting BCL-X(L) will most likely have widespread utility in cancer treatment and, instead of inhibiting multiple prosurvival BCL-2 family members, a BCL-X(L)-selective inhibitor would be expected to minimize the toxicity to normal tissues. We describe the use of a high-throughput screen to discover a new series of small molecules targeting BCL-X(L) and their structure-guided development by medicinal chemistry. The optimized compound, WEHI-539 (7), has high affinity (subnanomolar) and selectivity for BCL-X(L) and potently kills cells by selectively antagonizing its prosurvival activity. WEHI-539 will be an invaluable tool for distinguishing the roles of BCL-X(L) from those of its prosurvival relatives, both in normal cells and notably in malignant tumor cells, many of which may prove to rely upon BCL-X(L) for their sustained growth.


Assuntos
Antineoplásicos/farmacologia , Desenho de Fármacos , Proteína bcl-X/antagonistas & inibidores , Proteína bcl-X/química , Animais , Apoptose , Benzotiazóis/química , Linhagem Celular Tumoral , Cristalografia por Raios X , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Hidrazonas/química , Cinética , Camundongos , Modelos Químicos , Proteína de Sequência 1 de Leucemia de Células Mieloides , Ligação Proteica , Proteínas Proto-Oncogênicas c-bcl-2/genética
8.
Dis Model Mech ; 6(2): 521-9, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22996645

RESUMO

Anti-cancer drug development involves enormous expenditure and risk. For rapid and economical identification of novel, bioavailable anti-tumour chemicals, the use of appropriate in vivo tumour models suitable for large-scale screening is key. Using a Drosophila Ras-driven tumour model, we demonstrate that tumour overgrowth can be curtailed by feeding larvae with chemicals that have the in vivo pharmacokinetics essential for drug development and known efficacy against human tumour cells. We then develop an in vivo 96-well plate chemical screening platform to carry out large-scale chemical screening with the tumour model. In a proof-of-principle pilot screen of 2000 compounds, we identify the glutamine analogue, acivicin, a chemical with known activity against human tumour cells, as a potent and specific inhibitor of Drosophila tumour formation. RNAi-mediated knockdown of candidate acivicin target genes implicates an enzyme involved in pyrimidine biosynthesis, CTP synthase, as a possible crucial target of acivicin-mediated inhibition. Thus, the pilot screen has revealed that Drosophila tumours are glutamine-dependent, which is an emerging feature of many human cancers, and has validated the platform as a powerful and economical tool for in vivo chemical screening. The platform can also be adapted for use with other disease models, thus offering widespread applications in drug development.


Assuntos
Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Drosophila melanogaster/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Neoplasias/tratamento farmacológico , Animais , Benzamidas/farmacologia , Benzamidas/uso terapêutico , Disponibilidade Biológica , Proliferação de Células/efeitos dos fármacos , Citidina Trifosfato/biossíntese , Difenilamina/análogos & derivados , Difenilamina/farmacologia , Difenilamina/uso terapêutico , Drosophila melanogaster/citologia , Glutamina/metabolismo , Isoxazóis/farmacologia , Isoxazóis/uso terapêutico , Farmacogenética , Projetos Piloto
9.
J Biomol Screen ; 16(10): 1196-205, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22086725

RESUMO

Epigenetic aberrations are increasingly regarded as key factors in cancer progression. Recently, deregulation of histone acetyltransferases (HATs) has been linked to several types of cancer. Monocytic leukemia zinc finger protein (MOZ) is a member of the MYST family of HATs, which regulate gene expression in cell proliferation and differentiation. Deregulation of these processes through constitutively active MOZ fusion proteins gives rise to the formation of leukemic stem cells, rendering MOZ an excellent target for treating myeloid leukemia. The authors implemented a hit discovery campaign to identify small-molecule inhibitors of MOZ-HAT activity. They developed a robust, homogeneous assay measuring the acetylation of synthetic histone peptides. In a primary screening campaign testing 243 000 lead-like compounds, they identified inhibitors from several chemical classes. Secondary assays were used to eliminate assay-interfering compounds and prioritize confirmed hits. This study establishes a new high-throughput assay for HAT activity and could provide the foundation for the development of a new class of drugs for the treatment of leukemias.


Assuntos
Epigênese Genética/efeitos dos fármacos , Ensaios de Triagem em Larga Escala/métodos , Histona Acetiltransferases/metabolismo , Ativação Enzimática/efeitos dos fármacos , Histona Acetiltransferases/antagonistas & inibidores , Histona Acetiltransferases/genética , Humanos , Reprodutibilidade dos Testes , Bibliotecas de Moléculas Pequenas
10.
J Med Chem ; 54(6): 1914-26, 2011 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-21366295

RESUMO

ABT-737 and ABT-263 are potent inhibitors of the BH3 antiapoptotic proteins, Bcl-x(L) and Bcl-2. This class of putative anticancer agents invariantly contains an acylsulfonamide core. We have designed and synthesized a series of novel quinazoline-based inhibitors of Bcl-2 and Bcl-x(L) that contain a heterocyclic alternative to the acylsulfonamide. These compounds exhibit submicromolar, mechanism-based activity in human small-cell lung carcinoma cell lines in the presence of 10% human serum. This comprises the first successful demonstration of a quinazoline sulfonamide core serving as an effective benzoylsulfonamide bioisostere. Additionally, these novel quinazolines comprise only the second known class of Bcl-2 family protein inhibitors to induce mechanism-based cell death.


Assuntos
Apoptose , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Quinazolinas/síntese química , Sulfonamidas/síntese química , Proteína bcl-X/metabolismo , Animais , Ligação Competitiva , Linhagem Celular Tumoral , Cristalografia por Raios X , Ensaios de Seleção de Medicamentos Antitumorais , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Humanos , Camundongos , Modelos Moleculares , Proteína de Sequência 1 de Leucemia de Células Mieloides , Ligação Proteica , Proteínas Proto-Oncogênicas c-bcl-2/genética , Quinazolinas/química , Quinazolinas/farmacologia , Relação Estrutura-Atividade , Sulfonamidas/química , Sulfonamidas/farmacologia
11.
Antimicrob Agents Chemother ; 54(5): 1712-9, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20160053

RESUMO

The current treatment for leishmaniasis is based on chemotherapy, which relies on a handful of drugs with serious limitations, such as high cost, toxicity, and a lack of efficacy in regions of endemicity. Therefore, the development of new, effective, and affordable antileishmanial drugs is a global health priority. Leishmania synthesizes a range of mannose-rich glycoconjugates that are essential for parasite virulence and survival. A prerequisite for glycoconjugate biosynthesis is the conversion of monosaccharides to the activated mannose donor, GDP-mannose, the product of a reaction catalyzed by GDP-mannose pyrophosphorylase (GDP-MP). The deletion of the gene encoding GDP-MP in Leishmania led to a total loss of virulence, indicating that the enzyme is an ideal drug target. We developed a phosphate sensor-based high-throughput screening assay to quantify the activity of GDP-MP and screened a library containing approximately 80,000 lead-like compounds for GDP-MP inhibitors. On the basis of their GDP-MP inhibitory properties and chemical structures, the activities of 20 compounds which were not toxic to mammalian cells were tested against ex vivo amastigotes and in macrophage amastigote assays. The most potent compound identified in the primary screen (compound 3), a quinoline derivative, demonstrated dose-dependent activity in both assays (50% inhibitory concentration = 21.9 microM in the macrophage assay) and was shown to be nontoxic to human fibroblasts. In order to elucidate signs of an early structure-activity relationship (SAR) for this class of compounds, we obtained and tested analogues of compound 3 and undertook limited medicinal chemistry optimization, which included the use of a number of SAR probes of the piperazinyl aryl substituent of compound 3. We have identified novel candidate compounds for the design and synthesis of antileishmanial therapeutics.


Assuntos
Antiprotozoários/farmacologia , Desenho de Fármacos , Leishmania major/efeitos dos fármacos , Leishmaniose Cutânea/tratamento farmacológico , Nucleotidiltransferases/antagonistas & inibidores , Antiprotozoários/química , Células Cultivadas , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Fibroblastos/citologia , Fibroblastos/parasitologia , Humanos , Leishmania major/enzimologia , Leishmaniose Cutânea/parasitologia , Nucleotidiltransferases/metabolismo , Pirazóis/farmacologia , Quinolinas/farmacologia , Bibliotecas de Moléculas Pequenas , Tiadiazóis/farmacologia
12.
Antimicrob Agents Chemother ; 53(7): 2824-33, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19364854

RESUMO

High-throughput screening of 100,000 lead-like compounds led to the identification of nine novel chemical classes of trypanothione reductase (TR) inhibitors worthy of further investigation. Hits from five of these chemical classes have been developed further through different combinations of preliminary structure-activity relationship rate probing and assessment of antiparasitic activity, cytotoxicity, and chemical and in vitro metabolic properties. This has led to the identification of novel TR inhibitor chemotypes that are drug-like and display antiparasitic activity. For one class, a series of analogues have displayed a correlation between TR inhibition and antiparasitic activity. This paper explores the process of identifying, investigating, and evaluating a series of hits from a high-throughput screening campaign.


Assuntos
Antiparasitários/farmacologia , Avaliação Pré-Clínica de Medicamentos/métodos , NADH NADPH Oxirredutases/antagonistas & inibidores , Trypanosoma/efeitos dos fármacos , Animais , Antiparasitários/síntese química , Antiparasitários/química , Antiparasitários/uso terapêutico , Humanos , Microssomos Hepáticos/metabolismo , Estrutura Molecular , Relação Estrutura-Atividade , Tripanossomíase/tratamento farmacológico
13.
Bioorg Med Chem Lett ; 17(5): 1422-7, 2007 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-17194585

RESUMO

A high-throughput screening campaign of a library of 100,000 lead-like compounds identified 2-iminobenzimidazoles as a novel class of trypanothione reductase inhibitors. These 2-iminobenzimidazoles display potent trypanocidal activity against Trypanosoma brucei rhodesiense, do not inhibit closely related human glutathione reductase and have low cytotoxicity against mammalian cells.


Assuntos
Benzimidazóis/química , Benzimidazóis/farmacologia , Avaliação Pré-Clínica de Medicamentos/métodos , NADH NADPH Oxirredutases/antagonistas & inibidores , Animais , Benzimidazóis/toxicidade , Ligação Competitiva , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Química Combinatória , Humanos , Concentração Inibidora 50 , Relação Estrutura-Atividade , Trypanosoma brucei rhodesiense/efeitos dos fármacos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...