RESUMO
Research collaborations and licensing deals are critical for the discovery and development of life-saving drugs. This practice has been ongoing since the inception of the pharmaceutical industry. The current process of drug discovery and development is complex, regulated, and highly regimented, having evolved over time. Academia excels in the discovery of fundamental scientific concepts and biological processes, while industry excels in translational science and product development. Potential for collaboration exists at every step of the drug discovery and development continuum. This perspective walks through such collaborative activities, provides examples, and offers tips for potential collaborations.
Assuntos
Descoberta de Drogas , Indústria Farmacêutica , Humanos , História do Século XX , Comportamento Cooperativo , História do Século XXI , Desenvolvimento de Medicamentos , AcademiaRESUMO
The actinomycete Streptomyces platensis produces two compounds that display antibacterial activity: platensimycin and platencin. These compounds were discovered by the Merck Research Laboratories, and a complex insoluble production medium was reported. We have used this medium as our starting point in our studies. In a previous study, we developed a semi-defined production medium, i.e., PM5. In the present studies, by varying the concentration of the components of PM5, we were able to develop a superior semi-defined medium, i.e., PM6, which contains a higher concentration of lactose. Versions of PM6, containing lower concentrations of all components, were also found to be superior to PM5. The new semi-defined production media contain dextrin, lactose, MOPS buffer, and ammonium sulfate in different concentrations. We determined antibiotic production capabilities using agar diffusion assays and chemical assays via thin-layer silica chromatography and high-performance liquid chromatography. We reduced crude nutrient carryover from the seed medium by washing the cells with distilled water. Using these semi-defined media, we determined that addition of the semi-defined component soluble starch stimulated antibiotic production and that it and dextrin could both be replaced with glucose, resulting in the chemically defined medium, PM7.
Assuntos
Adamantano/metabolismo , Aminobenzoatos/metabolismo , Anilidas/metabolismo , Antibacterianos/metabolismo , Meios de Cultura/química , Streptomyces/crescimento & desenvolvimento , Streptomyces/metabolismo , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Testes de Sensibilidade MicrobianaRESUMO
The 70% aqueous methanolic extract of the Chinese plant Aristolochia manshuriensis was found to contain two novel substituted phenanthrene compounds, SCH 546909 (1), and another phenanthrene glycoside (2). The structures of 1 and 2 were established based on NMR studies. They were identified as inhibitors of the CDK2 enzyme. Compound 1 was found to be a potent inhibitor of the CDK2 enzyme with an IC50 of 140 nM, whereas compound 2 was found to be less active with an IC50 of >10 microM.
Assuntos
Antineoplásicos/farmacologia , Aristolochia/química , Quinase 2 Dependente de Ciclina/antagonistas & inibidores , Glicosídeos/química , Compostos Heterocíclicos de 4 ou mais Anéis/química , Extratos Vegetais/química , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Ativação Enzimática/efeitos dos fármacos , Glicosídeos/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Metanol/química , Estrutura Molecular , Água/químicaRESUMO
Several analogs of aristolochic acids were isolated and derivatized into their lactam derivatives to study their inhibition in CDK2 assay. The study helped to derive some conclusions about the structure-activity relation around the phenanthrin moiety. Semi-synthetic aristolactam 21 showed good activity with inhibition IC50 of 35 nM in CDK2 assay. The activity of this compound was comparable to some of the most potent synthetic compounds reported in the literature.
Assuntos
Ácidos Aristolóquicos/síntese química , Proteína Quinase CDC2/antagonistas & inibidores , Pirazóis/síntese química , Quinolinas/síntese química , Ácidos Aristolóquicos/química , Ácidos Aristolóquicos/farmacologia , Linhagem Celular Tumoral , Simulação por Computador , Cristalografia por Raios X , Humanos , Concentração Inibidora 50 , Modelos Moleculares , Estrutura Molecular , Pirazóis/química , Pirazóis/farmacologia , Quinolinas/química , Quinolinas/farmacologia , Relação Estrutura-AtividadeRESUMO
Four new caryophyllene derivatives, Sch 725432 (1), Sch 601253 (2), Sch 601254 (3), and Sch 725434 (4), were isolated from the fungal fermentation broth of Chrysosporium pilosum by reversed-phase HPLC purification. The structure elucidation of trioxygenated caryophyllenes 1-4 was accomplished on the basis of spectroscopic data interpretation. Sch 725434 (4) possesses a dihydrofuran-3-one ring, forming a tricyclic ring skeleton, which represents an unprecedented ring skeleton for the caryophyllene-type of sesquiterpenes. Compounds 1-4 were evaluated for their antifungal activity.
Assuntos
Chrysosporium/química , Saccharomyces cerevisiae/efeitos dos fármacos , Sesquiterpenos/isolamento & purificação , Antifúngicos/química , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Testes de Sensibilidade Microbiana , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Sesquiterpenos Policíclicos , Sesquiterpenos/química , Sesquiterpenos/farmacologiaRESUMO
Investigation of unexpected levels of impurities in Intron product has revealed the presence of low levels of impurities leached from the silicone tubing (Rehau RAU-SIK) on the Bosch filling line. In order to investigate the effect of these compounds (1a, 1b and 2) on humans, they were isolated identified and synthesized. They were extracted from the tubing by stirring in Intron placebo at room temperature for 72 h and were enriched on a reverse phase CHP-20P column, eluting with gradient aqueous ACN and were separated by HPLC. Structural elucidation of 1a, 1b and 2 by MS and NMR studies demonstrated them to be halogenated biphenyl carboxylic acids. The structures were confirmed by independent synthesis. Levels of extractable impurities in first filled vials of actual production are estimated to be in the range of 0.01-0.55 microg/vial for each leached impurity. Potential toxicity of these extractables does not represent a risk for patients under the conditions of clinical use.
Assuntos
Contaminação de Medicamentos , Interferon-alfa/química , Interferon-alfa/isolamento & purificação , Compostos Orgânicos/química , Silicones/química , Soluções Tampão , Química Farmacêutica , Cromatografia Líquida de Alta Pressão/métodos , Liofilização , Humanos , Concentração de Íons de Hidrogênio , Interferon alfa-2 , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas/métodos , Estrutura Molecular , Pós , Controle de Qualidade , Proteínas Recombinantes , Temperatura , Fatores de TempoRESUMO
A novel fungal secondary metabolite, Sch 213766 was isolated from the fungal fermentation broth of Chaetomium globosum as the chemokine receptor CCR-5 inhibitor and shown to be the methyl ester of the previously described tetramic acid Sch 210972 on the basis of UV, MS and NMR spectral data analyses. Sch213766 exhibited an IC(50) value of 8.6 muM in the CCR-5 receptor in vitro binding assay.
Assuntos
Antagonistas dos Receptores CCR5 , Chaetomium/metabolismo , Fármacos Anti-HIV/química , Fármacos Anti-HIV/isolamento & purificação , Fármacos Anti-HIV/metabolismo , Chaetomium/crescimento & desenvolvimento , Meios de Cultivo Condicionados/metabolismo , Fermentação , Éteres Metílicos/química , Éteres Metílicos/isolamento & purificação , Éteres Metílicos/metabolismo , Éteres Metílicos/farmacologiaRESUMO
Streptomyces platensis MA7327 is a bacterium producing interesting antibiotics, which act by the novel mechanism of inhibiting fatty acid biosynthesis. The antibiotics produced by this actinomycete are platensimycin and platencin plus some minor related antibiotics. Platensimycin and platencin have activity against antibiotic-resistant bacteria such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus; they also lack toxicity in animal models. Platensimycin also has activity against diabetes in a mouse model. We have been interested in studying the effects of primary metabolites on production of these antibiotics in our chemically defined production medium. In the present work, we tested 32 primary metabolites for their effect. They included 20 amino acids, 7 vitamins and 5 nucleic acid derivatives. Of these, only l-aspartic acid showed stimulation of antibiotic production. We conclude that the stimulatory effect of aspartic acid is due to its role as a precursor involved in the biosynthesis of aspartate-4-semialdehyde, which is the starting point for the biosynthesis of the 3-amino-2,4-dihydroxy benzoic acid portion of the platensimycin molecule.
Assuntos
Antibacterianos/isolamento & purificação , Ácido Aspártico/administração & dosagem , Streptomyces/metabolismo , Adamantano/isolamento & purificação , Aminoácidos/administração & dosagem , Aminoácidos/metabolismo , Aminobenzoatos/isolamento & purificação , Aminofenóis/isolamento & purificação , Anilidas/isolamento & purificação , Antibacterianos/biossíntese , Ácido Aspártico/química , Ácidos Nucleicos/administração & dosagem , Ácidos Nucleicos/metabolismo , Compostos Policíclicos/isolamento & purificação , Vitaminas/administração & dosagem , Vitaminas/metabolismoRESUMO
A new 5-alkenylresorcinol Sch725681 (1) was isolated and identified from the culture of an Aspergillus sp. The structure elucidation of 1 was accomplished based on extensive NMR spectroscopic analyses. Compound 1 showed inhibitory activity against Saccharomyces cerevisiae (PM503) and Candida albicans (C43) with MICs of 16 and 64 microg/ml, respectively.
Assuntos
Antifúngicos/isolamento & purificação , Aspergillus/metabolismo , Resorcinóis/isolamento & purificação , Antifúngicos/química , Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Resorcinóis/química , Resorcinóis/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacosRESUMO
A new hydrogenated azaphilone Sch725680 (1) was isolated and identified from the culture of an Aspergillus sp. The structure elucidation of 1 was achieved based on extensive NMR spectroscopic analyses. Compound 1 showed inhibitory activity against Saccharomyces cerevisiae (PM503) and Candida albicans (C43) with MICs of 8 and 64 microg/ml, respectively.
Assuntos
Antibacterianos/química , Antifúngicos/química , Aspergillus/química , Benzopiranos/química , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Aspergillus/metabolismo , Benzopiranos/isolamento & purificação , Benzopiranos/farmacologia , Candida albicans/efeitos dos fármacos , Fermentação , Testes de Sensibilidade Microbiana , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Saccharomyces cerevisiae/efeitos dos fármacos , Espectrometria de Massas por Ionização por Electrospray , Staphylococcus aureus/efeitos dos fármacosRESUMO
In recent years, large pharmaceutical companies have significantly reduced or eliminated the search for new therapeutic agents from natural sources. In spite of the many successes from natural product drug discovery, these companies have chosen to focus on compound libraries as the source of new lead compounds. Smaller biotechnology companies are continuing the search for novel natural products by developing and employing new and innovative approaches. This paper will describe some of these recent approaches to natural product drug discovery.:
RESUMO
A new microbial metabolite Sch 725424 (1) was isolated from the culture of Kitasatospora sp. The structure elucidation of 1 was accomplished based on NMR spectroscopic analyses as well as extensive structure elucidation of its dehydration product Sch 725428 (2). Compound 1 showed inhibitory activity against Staphylococcus aureus with MIC values 1-2 microg/ml, and also displayed weak antifungal activity against Saccharomyces cerevisiae (PM503) with an MIC 32 microg/ml.
Assuntos
Amidas/química , Amidas/metabolismo , Antibacterianos/biossíntese , Antibacterianos/química , Furanos/química , Furanos/metabolismo , Polienos/química , Polienos/metabolismo , Streptomycetaceae/metabolismo , Amidas/farmacologia , Antibacterianos/farmacologia , Antifúngicos/química , Antifúngicos/metabolismo , Antifúngicos/farmacologia , Fermentação , Furanos/farmacologia , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Estrutura Molecular , Polienos/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacosRESUMO
Two novel antibiotics, Sch 484129 (1) and Sch 484130 (2), were isolated from the fermentation broth of a fungal culture, which was identified as a Basidiomycete. The new antibiotics were obtained by ethyl acetate extraction followed by reversed phase HPLC purification. Structure elucidation of 1 and 2 was accomplished by spectroscopic data analyses. Derivatizations of the major component 1 were performed in order to provide definitive structural information. Both components were identified as glycolipids and displayed antifungal activity against Saccharomyces and Aspergillus strains.
Assuntos
Antifúngicos/isolamento & purificação , Basidiomycota/química , Glicolipídeos/isolamento & purificação , Monossacarídeos/isolamento & purificação , Antifúngicos/química , Antifúngicos/farmacologia , Aspergillus/efeitos dos fármacos , Basidiomycota/metabolismo , Fermentação , Glicolipídeos/química , Glicolipídeos/farmacologia , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Monossacarídeos/química , Monossacarídeos/farmacologia , Saccharomyces/efeitos dos fármacosRESUMO
Two novel antifungals SCH 643432 (1), and 2, were isolated from the fermentation broth of a fungus taxonomically classified as a Paecilomyces varioti. These compounds were separated from the fermentation broth filtrate by adsorption on a macroreticular resin XAD-16 (Amberlite). Purification and separation of the individual compounds were achieved by trituration of the extract with dichloromethane followed by preparative HPLC using reverse phase columns. Extensive FAB (Fast Atom Bombardment) and ESI (Electro Spray) mass spectrometric studies using fragmentation of various daughter ions, NMR experiments and degradative studies helped in elucidating the structure of compound 1. Compound 2 is an isomer of SCH 643432 (1). They were identified as straight chains peptides containing several amino acids such as alanine, aminoisobutyric acid, proline, leucine, glycine and arginine. The N-terminal is terminated in a previously identified beta-keto acid, 2-methyl 3-oxo tetradecanoic acid (MOTDA). Both compounds were active against Candida albicans, other Candidas, dermatophytes and Aspergillus (Geometric Mean MIC's 4.00, 2.59, 3.56, 11.31 and 4.49, 4.00, 5.66, 16.0 microg/ml, respectively for 1 and 2).
Assuntos
Antifúngicos/química , Fermentação , Fungos Mitospóricos/metabolismo , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Parede Celular/efeitos dos fármacosRESUMO
Platensimycin and platencin are compounds that were discovered at Merck Research Laboratories and have shown promising antibacterial activity. They are both produced in fermentation by the actinomycete Streptomyces platensis. Merck reported a crude, insoluble production medium to produce the antibiotics. To test the possible effects of different primary metabolites and inorganic compounds on the production of these antibiotics, a chemically-defined medium is needed. The effects that these compounds have on production could provide information about the precursors and biosynthetic pathway of the antibiotics. We have tested and developed a number of media with varying degrees of chemical definition and solubility using the Merck medium as our starting point. Our latest production medium, PM5, is soluble and semi-defined. It yields suitable production of the compounds, as shown by agar diffusion assays, bioautography and HPLC. The antibiotics were located in the extracellular broths and not in the mycelia.