RESUMO
The most important tools in killing and overcoming on the microbes and pathogens that cause diseases in medicine and/or in agriculture are the antibiotics. The discovery and synthesis of the microbial natural products or antibiotics has greatly developed genetically and biotechnologically quickly in the last decades. It is necessary to access this great genetic diversity by finding ways to increase the level of expression of these biosynthetic pathways. In this study, we carried out an improvement in the antibiotic production of weak Streptomyces graminofaciens strain NBR9 that has high lipid content; using Ultra-Violet irradiation mutagenesis. This strain was isolated from the Northern Region in the kingdom of Saudi Arabia and identified biochemically and confirmed genetically by sequencing of the 16S rRNA gene as Streptomyces graminofaciens NBR9; Accession No. (MN640578). The resultant mutant strain showed increasing in their antimicrobial activities. The methods and techniques used for the antibiotic extraction, purification, characterization and identification proved that the obtained antibiotic is same with antibiotic Carbomycin.
Assuntos
Vias Biossintéticas/genética , Microbiologia Industrial , Leucomicinas/biossíntese , Lipídeos/genética , Mutação/genética , Streptomyces/genética , Antibacterianos/biossíntese , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Leucomicinas/isolamento & purificação , Leucomicinas/farmacologia , Mutagênese , RNA Ribossômico 16S/genéticaRESUMO
Tylosin is a 16-membered macrolide broad-spectrum antibiotic that has an important role in veterinary medicine, active against Gram-positive and a restricted range of Gram-negative bacteria. We synthesized 15 types of tylosin-related derivatives by chemical modification and evaluated them against mastitis pathogens. Among them, 20-deoxy-20-{N-methyl-N-[1-(3-quinolyl)-1H-1,2,3-triazol-4-yl]methylamino}-5-O-mycaminosyltylonolide 2f and 20-deoxy-20-{N-benzyl-N-[1-(3-quinolyl)-1H-1,2,3-triazol-4-yl]methylamino}-5-O-mycaminosyltylonolide 2k were found to not only expand their antibacterial impact to include Gram-negative bacteria, such as Escherichia coli and Klebsiella pneumoniae, but also to retain or increase antibacterial activity against Gram-positive bacteria, such as Staphylococcus aureus and Streptococcus uberis in comparison with the parent tylosin.
Assuntos
Antibacterianos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Leucomicinas/farmacologia , Antibacterianos/síntese química , Antibacterianos/química , Leucomicinas/síntese química , Leucomicinas/química , Tilosina/farmacologiaRESUMO
BACKGROUND/AIMS: Macrolide antibiotics are effective drugs in chronic bronchiolitis and chronic rhinosinusitis with mucus hypersecretion. However, the mechanism of action is unclear. This study was designed to investigate the effect of azithromycin (AZM; 15-membered) and midecamycin acetate (MDM; 16-membered) on MUC5AC and MUC2 gene expression and secretion from human airway epithelial cells. The effects of the two macrolides on tumor necrosis factor-α (TNF-α) release were also examined. METHODS: Confluent NCI-H292 human mucoepidermoid airways epithelial cells were pretreated with AZM or MDM for 2 h and then stimulated with 200 nmol/l phorbol 12-myristate 13-acetate (PMA) for 8 h. The MUC5AC and MUC2 gene expression was measured by real-time quantitative RT-PCR. Total mucin in culture supernatants was measured using enzyme-linked lectin assay. Enzyme-linked immunosorbent assay was used to determine MUC5AC, MUC2 and TNF-α released by the cells. RESULTS: AZM and MDM attenuated PMA-induced MUC5AC and MUC2 gene and protein expression in NCI-H292 cells. They also suppressed PMA-mediated TNF-α in the cells. CONCLUSION: The present study demonstrates that AZM and MDM suppress the synthesis of mucin and TNF-α from human airway epithelial cells.
Assuntos
Antibacterianos/farmacologia , Azitromicina/farmacologia , Leucomicinas/farmacologia , Mucina-5AC/metabolismo , Mucina-2/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Células Epiteliais/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Mucina-5AC/genética , Mucina-2/genética , Ésteres de Forbol , Sistema Respiratório/citologiaRESUMO
A biotransformation approach using microbes as biocatalysts can be an efficient tool for the targeted modification of existing antibiotic chemical scaffolds to create previously uncharacterized therapeutic agents. By employing a recombinant Streptomyces venezuelae strain as a microbial catalyst, a reduced macrolide, 10,11-dihydrorosamicin, was created from rosamicin macrolide. Its chemical structure was spectroscopically elucidated, and the new rosamicin analog showed 2-4-fold higher antibacterial activity against two strains of methicillin-resistant Staphylococcus aureus compared with its parent rosamicin. This kind of biocatalytic approach is able to expand existing antibiotic entities and can also provide more diverse therapeutic resources.
Assuntos
Antibacterianos/metabolismo , Antibacterianos/farmacologia , Leucomicinas/metabolismo , Leucomicinas/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Streptomyces/metabolismo , Antibacterianos/química , Biotransformação , Leucomicinas/química , Macrolídeos/metabolismo , Macrolídeos/farmacologia , Testes de Sensibilidade Microbiana , Análise EspectralAssuntos
Antibacterianos/isolamento & purificação , Kitasamicina/análogos & derivados , Leucomicinas/isolamento & purificação , Micromonospora/metabolismo , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Engenharia Genética , Leucomicinas/farmacologia , Testes de Sensibilidade Microbiana , Micromonospora/genética , Ressonância Magnética Nuclear BiomolecularRESUMO
Design and synthesis of 16-membered macrolides modified at the C-12 and 13 positions are described. The compounds we report here have an arylalkylamino group attached to the C-12 position of the macrolactone. Both types of derivatives, 12,13-cyclic carbamates and non-carbamate analogues, were synthesized via 12-amino-13-hydroxy intermediates derived from 12,13-epoxide that was prepared by selective epoxidation at the C-12 and C-13 positions. 4'-Hydroxyl analogues were also prepared by acidic hydrolysis of a neutral sugar. These compounds were evaluated for in vitro antibacterial activity against respiratory tract pathogens. Some of these analogues exhibited an improved activity compared with the corresponding parent compound.
Assuntos
Antibacterianos/farmacologia , Carbamatos/síntese química , Carbamatos/farmacologia , Leucomicinas/síntese química , Leucomicinas/farmacologia , Miocamicina/síntese química , Miocamicina/farmacologia , Alquilação , Aminação , Antibacterianos/síntese química , Antibacterianos/química , Carbamatos/química , Hidroxilação , Cetolídeos/química , Leucomicinas/química , Viabilidade Microbiana/efeitos dos fármacos , Miocamicina/química , Estrutura MolecularRESUMO
In the present study, the effects of the macrolide antibiotic, midecamycin (MDM), on the Th2 cytokine response induced by the Staphylococcus aureus products, staphylococcal enterotoxin B (SEB), lipoteichoic acid (LTA), and peptidoglycan (PEG), was investigated in human peripheral blood mononuclear cells (PBMCs) from patients with atopic dermatitis (AD). MDM inhibited SEB-induced mRNA expression of the Th2 cytokines interleukin-4 (IL-4) and IL-5 in PBMCs from patients with AD. Furthermore, MDM also suppressed LTA-induced or PEG-induced IL-5 mRNA expression in these patients. Inhibition of mRNA expression by MDM correlated with the synthesis of cytokines in PBMCs, indicating that MDM controls Th2 cytokine production. In addition, S. aureus strains isolated from skin lesions of patients with AD were particularly susceptible to MDM compared with gentamicin, which is used widely in Japan as an antibiotic ointment combined with steroid for topical application in AD. These results suggest that topical administration of MDM might be beneficial in AD lesions infected with S. aureus.
Assuntos
Antibacterianos/farmacologia , Citocinas/biossíntese , Dermatite Atópica/imunologia , Leucomicinas/farmacologia , Staphylococcus aureus/fisiologia , Células Th2/efeitos dos fármacos , Adulto , Citocinas/genética , Enterotoxinas/antagonistas & inibidores , Regulação da Expressão Gênica , Gentamicinas/farmacologia , Humanos , Interleucina-4/biossíntese , Interleucina-4/genética , Interleucina-5/biossíntese , Interleucina-5/genética , Lipopolissacarídeos/antagonistas & inibidores , Macrolídeos/farmacologia , Peptidoglicano/farmacologia , RNA Mensageiro/biossíntese , Staphylococcus aureus/metabolismo , Ácidos Teicoicos/antagonistas & inibidores , Células Th2/imunologiaRESUMO
A total of 65 U. urealyticum cultures isolated from patients with chronic inflammatory diseases of the urogenital tract after their prolonged persistence of in the human body, were studied for sensitivity to medicinal preparations of different groups: tetracyclines (tetracycline and doxycycline), macrolids (erythomycin, clarithromycin, midecamycin, josamycin), quinolon (pefloxacin), amino glycoside (gentamicin), lincoamides (lincomycin, clindamycin). The majority of isolated U. urealyticun were highly sensitive to josamycin, clacide, doxycycline (89.2, 84.6, 76/9% respectively), and somewhat lesser number of these organisms were highly sensitive to midecamycin and pefloxacin (51.3 and 44.4% respectively). Among U. urealyticum strains circulating in the Moscow region some strains which persisted in patients with chronic inflammatory diseases of the urogenital tract for a long time were found to be resistant to erythromycin (23.1%), tetracycline (19.5%), and in very rare cases (1.6%) they were found to multiple drug resistance to all preparations under study. In view of the varying sensitivity of the clinical isolates of U. urealyticum to medicines and the presence of resistant forms in their population, the sensitivity of the isolated U. urealyticum should be determined in vitro prior to drug therapy.
Assuntos
Antibacterianos/farmacologia , Doenças Urogenitais Femininas/microbiologia , Doenças Urogenitais Masculinas , Infecções por Ureaplasma/microbiologia , Ureaplasma urealyticum/efeitos dos fármacos , Doença Crônica , Claritromicina/farmacologia , Doxiciclina/farmacologia , Doenças Urogenitais Femininas/patologia , Humanos , Inflamação/patologia , Josamicina/farmacologia , Leucomicinas/farmacologia , Testes de Sensibilidade Microbiana , Pefloxacina/farmacologia , Infecções por Ureaplasma/patologia , Ureaplasma urealyticum/isolamento & purificaçãoRESUMO
The synthesis and biological evaluation of sixteen-membered macrolides modified at the C-3 position are described. 3-Epi-leucomycin A7 (9), 3-O-acyl-3-epi-leucomycin A7 analogues (11a-11e), 3-O-acylleucomycin A7 analogues (13b-13e) and 3-O-methylleucomycin analogues (16a, 16b and 22) were synthesized via fully protected intermediates (7, 5a, 5b and 20). After appropriate modification, subsequent deprotections were performed to furnish a variety of leucomycin analogues. Methylation of the 3-hydroxyl group was found to improve the pharmacoprofile of leucomycin antibiotics. 3-O-Methylrokitamycin (16b) showed enhanced antibacterial activity in vitro and 3,3''-di-O-methyl-4''-O-(3-methylbutyl)leucomycin V (22) exhibited improved metabolic stability in rat plasma in vitro.
Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Leucomicinas/síntese química , Leucomicinas/farmacologia , Antibacterianos/química , Haemophilus influenzae/efeitos dos fármacos , Leucomicinas/química , Metilação , Testes de Sensibilidade Microbiana , Micrococcus luteus/efeitos dos fármacos , Estrutura Molecular , Moraxella catarrhalis/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Streptococcus pneumoniae/efeitos dos fármacosRESUMO
Glucuronide derivatives of CBI-bearing CC-1065 analogues have been synthesized, and their cytotoxicities tested against U937 leukemia cells. The new compounds show potent antitumor activity in vitro. Compounds 1 and 2, and their corresponding glucuronides 3 and 4 have IC(50) values of 0.6, 0.1, 1.4 and 0.6 nM, respectively. Glucuronide 3 is approximately 2-fold less toxic than its hydroxyl counterpart 1, and glucuronide 4 is approximately 6-fold less toxic than its hydroxyl counterpart 2. Glucuronides 3 and 4 may have limited use in the ADEPT approach. However, they may be used as antitumor agents in a conventional way.
Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Indóis , Leucomicinas/síntese química , Leucomicinas/farmacologia , DNA de Neoplasias/biossíntese , Ensaios de Seleção de Medicamentos Antitumorais , Duocarmicinas , Glucuronidase/metabolismo , Glucuronídeos/síntese química , Glucuronídeos/farmacologia , Humanos , Indicadores e Reagentes , Espectroscopia de Ressonância Magnética , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
CC-1065 analogues bearing different DNA-binding subunits were synthesized. A terminal C5-NO2 and -F moiety at the DNA-binding subunit increased the drug's potency and antitumor efficacy. A C5-OCH3 reduced the potency and antitumor efficacy. Compound (+/-)-7, bearing a trans double bond, had increased antitumor efficacy. A preliminary toxicity study indicated that terminal C5-OCH3 and -acetamido moieties at the DNA-binding subunit caused delayed death in mice.
Assuntos
Antineoplásicos/síntese química , DNA/química , Indóis/síntese química , Leucomicinas/síntese química , Animais , Antineoplásicos/farmacologia , Antineoplásicos/toxicidade , Ensaios de Seleção de Medicamentos Antitumorais , Duocarmicinas , Indóis/farmacologia , Indóis/toxicidade , Leucomicinas/química , Leucomicinas/farmacologia , Leucemia L1210/tratamento farmacológico , Camundongos , Estereoisomerismo , Relação Estrutura-Atividade , Taxa de Sobrevida , Testes de Toxicidade AgudaRESUMO
The cytocidal effect of seven macrolide antibiotics on human periodontal ligament fibroblasts (Pel cells) was studied. Pel cells were exposed for 48 h to erythromycin (EM), clarithromycin (CAM), roxithromycin (RXM), azithromycin (AZM), josamycin (JM), midecamycin (MDM), and rokitamycin (RKM), and allowed to form colonies. The cytocidal effect of the macrolides was measured as a decrease in colony-forming efficiency and was found to increase with the concentration. To obtain a quantitative measure of the cytocidal effect, the LD50, i.e. the concentration that decreases colony-forming efficiency 50% relative to control cells, was extrapolated from the concentration-response curves. The rank of the macrolides according to their cytocidal effect (LD50) was RKM > RXM > CAM > AZM > JM > MDM approximately EM. RKM, RXM, CAM, AZM, and JM were at least 1.7-12.2 times more cytocidal than MDM or EM. When extrapolated from the concentration-response curves, the relative survival of the Pel cells exposed to each of the macrolides at the MIC90 concentrations for periodontopathic bacteria was estimated to be: > or = 53.8% for RKM, > or = 92.7% for RXM, > or = 94.6% for CAM, > or = 97.1% for AZM, and > or = 86.2% for EM. The effect of the antibiotics on the mRNA expression of alkaline phosphatase (ALP) and type I procollagen (COL) was examined in Pel cells exposed for 48 h to RXM, CAM, AZM, and EM, which exhibited strong, moderate, and weak cytocidal activity. The constitutive levels of both ALP and COL mRNA were retained in cells exposed to RXM at < or = 3 microM, CAM at < or = 10 microM, and AZM or EM at < or = 3 microM. The MIC90 against periodontopathic bacteria is < or = 4.8 microM for RXM, 5.3 microM for CAM, 2.7 microM for AZM, and 21.8 microM for EM. These results suggest that topical administration of CAM or AZM to the gingival crevice at their MIC90 concentration for periodontopathic bacteria would have little adverse effect on the growth and differentiation of the periodontal ligament. It is important to note, however, that these findings have yet to be extrapolated to in vivo conditions.
Assuntos
Antibacterianos/farmacologia , Fibroblastos/efeitos dos fármacos , Miocamicina/análogos & derivados , Ligamento Periodontal/efeitos dos fármacos , Fosfatase Alcalina/efeitos dos fármacos , Fosfatase Alcalina/genética , Antibacterianos/administração & dosagem , Apoptose , Azitromicina/administração & dosagem , Azitromicina/farmacologia , Sobrevivência Celular , Células Cultivadas , Claritromicina/administração & dosagem , Claritromicina/farmacologia , Colágeno Tipo I/efeitos dos fármacos , Colágeno Tipo I/genética , Relação Dose-Resposta a Droga , Eritromicina/administração & dosagem , Eritromicina/farmacologia , Expressão Gênica/efeitos dos fármacos , Humanos , Josamicina/administração & dosagem , Josamicina/farmacologia , Dose Letal Mediana , Leucomicinas/administração & dosagem , Leucomicinas/farmacologia , Miocamicina/administração & dosagem , Miocamicina/farmacologia , Ligamento Periodontal/citologia , RNA Mensageiro/efeitos dos fármacos , Roxitromicina/administração & dosagem , Roxitromicina/farmacologia , Estatística como Assunto , Fatores de TempoRESUMO
The duocarmycins and (+)-CC-1065 are amongst the most potent antitumour antibiotics discovered to date and yet have not progressed into the clinic. The natural products are extremely stable to nucleophilic attack until bound to their DNA target and are not substrates for any other biological nucleophile. The mechanism for this target activation of the duocarmycins is discussed with relation to both an acid-catalyzed activation and a binding-induced conformational change leading to ground state destabilization. It is suggested that targeting of the duocarmycins to their site of action in a tumour may be more important than introducing systemically-activated prodrugs as the natural product itself can be considered to be a type of prodrug, activated only on binding to its targets. Methods that have been used to target CC-1065 and the duocarmycins are reviewed as well as efforts towards systemically activated prodrugs. A simple analysis of the approaches that could be taken to vary the structure for targeting is suggested.
Assuntos
Antibióticos Antineoplásicos/química , Antineoplásicos Alquilantes/química , DNA/química , Indóis , Pró-Fármacos/química , Animais , Antibióticos Antineoplásicos/farmacologia , Antineoplásicos Alquilantes/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Estabilidade de Medicamentos , Duocarmicinas , Leucomicinas/química , Leucomicinas/farmacologia , Leucemia L1210 , Pró-Fármacos/farmacologia , Pirróis/química , Pirróis/farmacologia , Pirrolidinonas/química , Pirrolidinonas/farmacologia , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
The effect of the macrolide antibiotics, clarithromycin, midecamycin acetate and josamycin, on the generation of Th1- and Th2-type cytokines by mitogen-stimulated human T lymphocytes was compared with that of fosfomycin. The following results were obtained. These drugs demonstrated potent inhibitory activity on the release and gene expression of TNF-alpha and IL-2. Their inhibitory effect on IFN-alpha, IL-4, IL-5, IL-6 was less marked. The release of IL-10 was poorly suppressed. Clarithromycin had the most potent inhibitory effect of the drugs used. The present results suggested that anti-bacterial agents might modify the host's immunological response by interfering with the activity of T helper cells.
Assuntos
Antibacterianos/farmacologia , Citocinas/biossíntese , Células Th1/imunologia , Células Th2/imunologia , Células Cultivadas , Claritromicina/farmacologia , Fosfomicina/farmacologia , Humanos , Interleucina-2/biossíntese , Interleucina-2/genética , Josamicina/farmacologia , Leucomicinas/farmacologia , Ativação Linfocitária , RNA Mensageiro/análise , Células Th1/efeitos dos fármacos , Células Th2/efeitos dos fármacos , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genéticaRESUMO
The solution-phase, parallel synthesis and evaluation of a library of 132 (+)-1,2,9,9a-tetrahydrocyclopropa[c]benz[e]indol-4-one (CBI) analogues of CC-1065 and the duocarmycins containing dimeric monocyclic, bicyclic, and tricyclic heteroaromatic replacements for the DNA-binding domain are described. This systematic study revealed clear trends in the structural requirements for observation of potent cytotoxic activity and DNA alkylation efficiency, the range of which spans a magnitude of > or =10 000-fold. Combined with related studies, these results highlight that the role of the DNA-binding domain goes beyond simply providing DNA-binding selectivity and affinity (10-100-fold enhancement in properties), consistent with the proposal that it contributes significantly to catalysis of the DNA alkylation reaction accounting for as much as an additional 1000-fold enhancement in properties.
Assuntos
Antibióticos Antineoplásicos/síntese química , Antibióticos Antineoplásicos/farmacologia , Leucomicinas/síntese química , Alquilantes/síntese química , Alquilantes/química , Alquilantes/farmacologia , Alquilação/efeitos dos fármacos , Antibióticos Antineoplásicos/química , Sítios de Ligação , Técnicas de Química Combinatória , Ciclopropanos/química , DNA/metabolismo , DNA Viral/efeitos dos fármacos , DNA Viral/metabolismo , Duocarmicinas , Indóis/química , Concentração Inibidora 50 , Leucomicinas/química , Leucomicinas/farmacologia , Pirróis/síntese química , Pirróis/química , Pirróis/farmacologia , Pirrolidinonas/síntese química , Pirrolidinonas/química , Pirrolidinonas/farmacologia , Vírus 40 dos Símios/genética , Relação Estrutura-AtividadeRESUMO
OBJECTIVE: To compare the in vitro activity of midecamycin diacetate to that of five other macrolides (erythromycin, clarithromycin, roxithromycin, azithromycin, and josamycin) and of clindamycin against 146 clinical isolates of Streptococcus pyogenes, with regard to three different phenotypes of erythromycin resistance. METHODS: Susceptibility pattern and resistance phenotype were determined by disk diffusion method and double disk test. Minimal inhibitory concentrations of antibiotics were obtained by the agar dilution method and evaluated according to the recommendations of the 'Comité de l'Antibiogramme de la Société Française de Microbiologie' (CA-SFM). The major determinants of erythromycin resistance in S. pyogenes (ermB, ermTR and mefA genes) were investigated by specific amplification protocols. RESULTS: Most of the isolates of S. pyogenes collected during 1995-99 were susceptible to midecamycin (93.8%), erythromycin (90.4%), clarithromycin (93.2%), roxithromycin (91.8%), azithromycin (88.4%), josamycin (94.5%), and clindamycin (94.5%). According to the CA-SFM criteria, 132 of the 146 isolates studied were susceptible to erythromycin (MICs < or = 1 mg/L), four were intermediate (MICs 2-4 mg/L), and 10 were resistant (MICs > 4 mg/L). Only nine isolates were midecamycin resistant (MICs > 4 mg/L), and the others were susceptible. The increased activity of midecamycin (MIC90 < or = 0.06 mg/L), as compared to erythromycin (MIC90 = 0.5 mg/L) and to other 14- or 15-membered macrolides, was related to the absence of the ermB determinant in seven isolates which displayed an efflux phenotype (five isolates) or an inducible resistance phenotype due to an ermTR determinant (two isolates). CONCLUSION: Midecamycin diacetate is active against most S. pyogenes strains isolated in France and may represent an attractive alternative to the treatment of streptococcal infections due to resistant isolates with efflux of erythromycin.
Assuntos
Antibacterianos/farmacologia , Leucomicinas/farmacologia , Streptococcus pyogenes/efeitos dos fármacos , Azitromicina/farmacologia , Claritromicina/farmacologia , Clindamicina/farmacologia , Contagem de Colônia Microbiana , Resistência Microbiana a Medicamentos , Eritromicina/farmacologia , França , Humanos , Josamicina/farmacologia , Testes de Sensibilidade Microbiana , Fenótipo , Reação em Cadeia da Polimerase , Roxitromicina/farmacologia , Streptococcus pyogenes/crescimento & desenvolvimentoRESUMO
The synthesis and examination of a novel class of reversed CPyI analogues of CC-1065 and the duocarmycins are described. Capable of a unique metal cation activation of DNA alkylation, these agents allowed the effects of the DNA binding domain (10(4)-fold increase in DNA alkylation rate and efficiency) to be partitioned into two components: that derived from enhanced DNA binding affinity and selectivity (10-80-fold) and that derived from a contribution to catalysis (250-5000-fold). In addition, the reversed enantiomeric selectivity of these sequence selective DNA alkylating agents provides further strong support for a previously disclosed model where it is the noncovalent binding selectivity of the compounds, and not the alkylation subunit or the source of catalysis, that controls the DNA alkylation selectivity.
Assuntos
Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacologia , Indóis/química , Indóis/farmacologia , Pirróis/química , Pirróis/farmacologia , Quinolonas/química , Quinolonas/farmacologia , Alquilantes/síntese química , Alquilantes/química , Alquilantes/farmacologia , Animais , Antibióticos Antineoplásicos/síntese química , Catálise , DNA/química , DNA/metabolismo , Duocarmicinas , Indóis/síntese química , Cinética , Leucomicinas/química , Leucomicinas/farmacologia , Leucemia L1210/tratamento farmacológico , Pirróis/síntese química , Quinolonas/síntese química , EstereoisomerismoRESUMO
The synthesis and evaluation of a series of C3-substituted 1,2,9,9a-tetrahydrocyclopropa[c]benz[e]indol-4-one (CBI) analogues of the CC-1065 and duocarmycin alkylation subunits are detailed, including methyl and the full series of halogens. Introduction of the key substituent was accomplished through directed metalation of the seco-CBI core followed by reaction of the resultant aryllithium with an appropriate electrophile. C3-Bromo and iodo substituents were only effectively installed on the hindered aryllithium intermediate using a novel halogen source, 1-bromo- and 1-iodophenylacetylene, that should prove generally useful beyond the studies we describe. X-ray crystal structures of the series show substantial distortion in the vinylogous amide due to unfavorable steric interactions between the C3-substituent and the N(2)-carbamate. In the halogen series, the N2-C2a bond length and the torsional angle chi(1) smoothly increase with the increasing size of the C3 substituent indicative of decreasing vinylogous amide conjugation through the series (H > F > Cl > Br > I). Unlike N-Boc-CBI, this series of substituted CBI analogues proved remarkably reactive toward solvolysis even at pH 7, where the reaction is uncatalyzed and the reactivity order (I > Br > Cl > F > H) follows a trend consistent with the extent of vinylogous amide conjugation and stabilization. The implications of these observations on the source of catalysis for the DNA alkylation reaction of the natural products are discussed.
Assuntos
Antibióticos Antineoplásicos/síntese química , Indóis , Leucomicinas/síntese química , Pirróis/síntese química , Pirrolidinonas/síntese química , Alquilação , Animais , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacologia , Cromatografia Líquida de Alta Pressão , Cristalografia por Raios X , Duocarmicinas , Humanos , Leucomicinas/química , Leucomicinas/farmacologia , Leucemia L1210/tratamento farmacológico , Espectroscopia de Ressonância Magnética , Conformação Molecular , Pirróis/química , Pirróis/farmacologia , Pirrolidinonas/química , Pirrolidinonas/farmacologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrofotometria UltravioletaRESUMO
A series of CBI analogues of the duocarmycins and CC-1065 exploring substituent effects within the first indole DNA binding subunit are detailed. Substitution at the indole C5 position led to cytotoxic potency enhancements that are > or =1000-fold, providing simplified analogues containing a single DNA binding subunit that are more potent (IC(50)=2-3 pM) than CBI-TMI, duocarmycin SA, or CC-1065.
Assuntos
Antibióticos Antineoplásicos/síntese química , DNA/química , Leucomicinas/síntese química , Pirróis/síntese química , Pirrolidinonas/síntese química , Adenina/análogos & derivados , Adenina/química , Adenina/metabolismo , Alquilação , Antibióticos Antineoplásicos/farmacologia , Sítios de Ligação/fisiologia , DNA/metabolismo , Adutos de DNA/química , Adutos de DNA/metabolismo , Duocarmicinas , Humanos , Indóis/química , Concentração Inibidora 50 , Leucomicinas/química , Leucomicinas/metabolismo , Leucomicinas/farmacologia , Conformação de Ácido Nucleico , Pirróis/farmacologia , Pirrolidinonas/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
[reaction in text] A novel photocyclization of tosylstilbenoids is used in the preparation of a cyclopropafuroindolone analogue of the DNA-alkylating unit of the antitumor compound CC-1065.