RESUMO
The hybridising potential of anhydrohexitol nucleoside analogues (HNAs) is well documented, but tedious synthesis of the monomers hampers their development. In a search for better analogues, the synthesis of two new methylated anhydrohexitol congeners 1 and 2 was accomplished and the physico-chemical properties of their respective oligomers were evaluated. Generally, oligonucleotides (ONs) containing the 3'-O-methyl derivative 1 showed a small increase in thermal stability towards complementary sequences as compared to HNA. Compared to the altritol modification, 3'-O-methylation seems to cause a small decrease in thermal stability of duplexes, especially when targeting RNA. These results suggest the possibility of derivatisation of the 3'-hydroxyl group of altritol-containing congeners without significantly affecting the thermal stability of the duplexes. The methyl glycosidic analogues 2 likewise increased the affinity for RNA in comparison with well-known HNA, while at the same time being economically more favorable monomers. However, homopolymers of 2 displayed self-pairing, but not so homopolymers of 1. Upon incorporation of the hexitols within RNA sequences in an effort to induce a beneficial pre-organised structure, the positive effect of the 3'-O-methyl derivative 1 proved larger than that of 2.
Assuntos
Hibridização de Ácido Nucleico/métodos , Oligonucleotídeos Antissenso/química , Oligonucleotídeos Antissenso/metabolismo , Álcoois Açúcares/metabolismo , Metilação , Oligonucleotídeos Antissenso/síntese química , Polirribonucleotídeos/metabolismo , RNA/metabolismo , Estabilidade de RNARESUMO
BACKGROUND: Synthetic nucleic acid analogues with a conformationally restricted sugar-phosphate backbone are widely used in antisense strategies for biomedical and biochemical applications. The modified backbone protects the oligonucleotides against degradation within the living cell, which allows them to form stable duplexes with sequences in target mRNAs with the aim of arresting their translation. The biologically most active antisense oligonucleotides also trigger cleavage of the target RNA through activation of endogenous RNase H. Systematic studies of synthetic oligonucleotides have also been conducted to delineate the origin of the chirality of DNA and RNA that are both composed of D-nucleosides. RESULTS: Hexitol nucleic acids (HNA) are the first example of oligonucleotides with a six-membered carbohydrate moiety that can bind strongly and selectively to complementary RNA oligomers. We present the first high resolution nuclear magnetic resonance structure of a HNA oligomer bound to a complementary RNA strand. The HNA-RNA complex forms an anti-parallel heteroduplex and adopts a helical conformation that belongs to the A-type family. Possibly, due to the rigidity of the rigid chair conformation of the six-membered ring both the HNA and RNA strand in the duplex are well defined. The observed absence of end-fraying effects also indicate a reduced conformational flexibility of the HNA-RNA duplex compared to canonical dsRNA or an RNA-DNA duplex. CONCLUSIONS: The P-P distance across the minor groove, which is close to A-form, and the rigid conformation of the HNA-RNA complex, explain its resistance towards degradation by Rnase H. The A-form character of the HNA-RNA duplex and the reduced flexibility of the HNA strand is possibly responsible for the stereoselectivity of HNA templates in non-enzymatic replication of oligonucleotides, supporting the theory that nucleosides with six-membered rings could have existed at some stage in molecular evolution.
Assuntos
DNA/química , Conformação de Ácido Nucleico , Ácidos Nucleicos Heteroduplexes/química , Oligorribonucleotídeos/química , RNA de Cadeia Dupla/química , RNA/química , Álcoois Açúcares/química , Sequência de Bases , Modelos Moleculares , SoluçõesRESUMO
The human N-acetylglucosaminyltransferase I gene was introduced in the genome of Trichoderma reesei strain VTT-D-80133. Expression was studied after induction from the cellobiohydrolase I promoter. Successful in vivo transfer of GlcNAc was demonstrated by analyzing the neutral N-glycans which were synthesized on cellobiohydrolase I. Final proof of the formation of GlcNAcMan5GlcNAc2 was obtained by NMR analysis.
Assuntos
N-Acetilglucosaminiltransferases/metabolismo , Polissacarídeos/biossíntese , Trichoderma/fisiologia , Sequência de Carboidratos , Celulase/genética , Celulose 1,4-beta-Celobiosidase , Clonagem Molecular , Humanos , Espectroscopia de Ressonância Magnética , Dados de Sequência Molecular , N-Acetilglucosaminiltransferases/genética , Oligossacarídeos/química , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Regiões Promotoras Genéticas , Proteínas Recombinantes/metabolismo , Esporos Fúngicos , Trichoderma/metabolismoRESUMO
Several water-soluble ester derivatives of acyclovir [9-[(2-hydroxyethoxy)methyl]guanine], i.e., the 2'-O-glycyl-, 2'-O-alpha-alanyl-, 2'-O-beta-alanyl- and 2'-O-3-carboxypropionyl esters, were synthesized and evaluated for their antiviral activity in cell culture. The compounds were all prepared directly from acyclovir by application of the usual esterification methods with the appropriate acyl precursors and isolated as their hydrochloride or sodium salts. When assayed in primary rabbit kidney cell cultures against various herpes simplex virus type 1 and type 2 strains, the four acyclovir esters proved almost as active as acyclovir itself, suggesting that they were readily hydrolyzed to release the parent compound.
Assuntos
Aciclovir/análogos & derivados , Aciclovir/farmacologia , Animais , Linhagem Celular , Rim , Coelhos , Simplexvirus/efeitos dos fármacos , Vírus da Estomatite Vesicular Indiana/efeitos dos fármacosRESUMO
5'-O-Phosphonomethylation of different pyrimidine 2',3'-dideoxynucleosides was accomplished by reaction of the latter with diethyl [(p-toylsulfonyl)oxy]methanephosphonate (1) in the presence of sodium hydride. The base-phosphonomethylated (15-19) and sugar-phosphonomethylated (8-12) derivatives could be readily distinguished by 1H and 13C NMR and MS analysis. Protection of the uracil or thymine residue with a N3-benzoyl group failed to prevent base modification. However, O4-methyl-protected 2',3'-dideoxyuridine readily afforded the 5'-O-phosphonomethylated derivative 12, which was converted to both the 2',3'-didoxyuridine analogue 27 and the 2',3'-dideoxycytidine counterpart 29. The 5'-O-phosphonomethyl derivatives of 3'-deoxythymidine (23), 2',3'-dideoxyuridine, (27), 2',3'-dideoxycytidine (29), 3'-O-methylthymidine (26), and 3'-amino-3'-deoxythymidine (28) did not show an appreciable anti-HIV activity in MT-4 cells. In contrast, the 5'-O-phosphonomethyl derivatives of 3'-deoxy-3'-fluorothymidine (24) and 3'-azido-3'-deoxythymidine (25) inhibited HIV-1 cytopathogenicity by 50% at a concentration of approximately 1 microM.
Assuntos
Antivirais/síntese química , Didesoxinucleosídeos/síntese química , Compostos Organofosforados/síntese química , Antivirais/farmacologia , Fenômenos Químicos , Química , Didesoxinucleosídeos/uso terapêutico , HIV-1/efeitos dos fármacos , Humanos , Compostos Organofosforados/uso terapêutico , Inibidores da Transcriptase Reversa , Relação Estrutura-AtividadeRESUMO
The clinical use of the potent, wide-spectrum aminoglycoside antibiotics is limited by oto- and nephrotoxicities. The latter is related to the binding of these polycationic drugs to negatively charged phospholipids and to the subsequent inhibition of lysosomal phospholipases. In order to explore the influence of a modification of the hydrophobic/hydrophilic balance at a specific site of an aminoglycoside, kanamycin B has been chemically modified in position 6" by substitution of the hydroxyl group with a halogen atom (or a pseudohalogen group), or an amino, an amido, a thioalkyl, or an alkoxy group, each series containing increasingly bulkier chains. Examination of the antibacterial activity of the synthesized compounds revealed a negative correlation between the size of the 6"-substituent and the antibacterial activity against kanamycin B sensitive Gram-positive and -negative organisms. Only derivatives with small substituents in position 6", namely chloro, bromo, azido, amino, methylcarbamido, acetamido, methylthio, methylsulfinyl, O-methyl, O-ethyl, and O-isopropyl, showed acceptable activity (geometric mean of minimum inhibitory concentrations for Gram-negative strains less than or equal to 2.5 mg/L; value for kanamycin B, 0.5 mg/L). In vitro toxicological evaluation of all derivatives and computer-aided conformational analysis of selected compounds inserted in a phosphatidylinositol monolayer are presented in the following paper in this issue.
Assuntos
Canamicina/análogos & derivados , Canamicina/síntese química , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Indicadores e Reagentes , Canamicina/química , Canamicina/farmacologia , Resistência a Canamicina , Testes de Sensibilidade Microbiana , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
Substitution of the C-1 atom in the 2-deoxystreptamine moiety of gentamicin C2, a broad-spectrum aminoglycoside antibiotic, by an axial hydroxymethyl group has been reported to confer protection against most clinically important bacterial enzymes inactivating aminoglycosides, while simultaneously reducing the nephrotoxic potential of this drug. We report here on a similar modification of kanamycin B. Microbiological evaluation, however, revealed no useful protection, as established by the almost complete lack of activity of 1-C-(hydroxymethyl)kanamycin B against an array of organisms producing defined types of aminoglycoside-inactivating enzymes and against which 1-C-(hydroxymethyl)gentamicin C2 and amikacin (1-N-[(S)-2-hydroxy-4-aminobutyryl]kanamycin A) are active. Moreover, toxicological evaluation, based on the in vitro measurement of the drug inhibitory potential toward lysosomal phospholipases, a predictive test of the intrinsic nephrotoxic potential of aminoglycosides, showed not decreased but rather increased toxicity. Comparative conformational analysis of the interactions of the drug with a phosphatidylinositol monolayer explained the lack of protective effect, since no significant change of the mode of insertion of the derivative in this monolayer was detected compared to that of kanamycin B. Combination of a 1-C-(hydroxymethyl) substituent with a 6"-chloro, 6"-acetamido substituent resulted in a partial improvement of the toxicological behavior with no loss of activity for the 6"-chloro and the 6"-azido derivatives, but not to the extent of obtaining better derivatives than kanamycin B itself. We, therefore, suggest that the advantages of an axial hydroxymethyl substituent at C-1 are probably restricted to the gentamicin family and do not extend to kanamycins. It might be concluded that the structural differences between gentamicins and kanamycins play an important, still undescribed role both in their effective recognition by aminoglycoside-inactivating enzymes, which are responsible for most of the clinically important cases of resistance to aminoglycosides, and also in the interactions with phospholipids, which in turn cause nephrotoxicity.
Assuntos
Canamicina/análogos & derivados , Canamicina/síntese química , Bactérias Gram-Positivas/efeitos dos fármacos , Indicadores e Reagentes , Canamicina/química , Canamicina/farmacologia , Canamicina/toxicidade , Testes de Sensibilidade Microbiana , Modelos Moleculares , Conformação Molecular , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
In a companion paper (previous paper in this issue), we report on the synthesis and microbiological evaluation of new derivatives of the aminoglycoside antibiotic kanamycin B carrying substitutions in 6" (halogeno, or amino, amido, thioalkyl, and alkoxy groups, each series with increasingly bulkier chains). These modifications were intended to potentially modulate the interactions of kanamycin B with phospholipids since these are related to inhibition of lysosomal phospholipase activities and lysosomal phospholipidosis, an early and predictive index of the nephrotoxic potential of aminoglycosides. The new derivatives were therefore examined for inhibitory potency in vitro toward lysosomal phospholipase A1 acting on phosphatidylcholine included in negatively charged liposomes. No simple correlation was observed between the nature or the size of the 6''-substituent and the inhibitory potencies of the corresponding derivatives, although certain groups (diethylamino, isopropylthio) caused a significant increase in inhibitory potency, whereas an N-acetyl-N-methylamino substituent had the opposite effect. 6''-Deoxy-6''-chlorokanamycin B, however, was the only derivative showing both a decrease (albeit limited) of inhibitory potency toward phospholipase A1 associated with the maintenance of a satisfactory microbiological activity (actually equal or slightly better than that of kanamycin B). Computer-aided conformational analysis showed that this chloro substituent did not allow the molecule to insert itself very differently compared to kanamycin B or 6''-deoxykanamycin B in a monolayer of phosphatidylinositol, all three drugs adopting an orientation largely parallel to the hydrophobic-hydrophilic interface and being largely "embedded" in the bilayer at that level. In contrast, the N-acetyl-N-methylamino and isopropylthio substituents caused the corresponding derivatives to adopt an orientation largely perpendicular to the interface, because of the attraction of this substituent, and therefore of the 3''-amino sugar moiety of kanamycin B into the hydrophobic domain of the monolayer, whereas the opposite part of the drug (2',6'-diamino sugar) protruded into the aqueous phase. No simple correlation, however, could be drawn between these changes of conformation and the relative inhibitory potencies of the derivatives.
Assuntos
Canamicina/análogos & derivados , Canamicina/síntese química , Gráficos por Computador , Indicadores e Reagentes , Canamicina/química , Canamicina/farmacologia , Lisossomos/enzimologia , Modelos Moleculares , Conformação Molecular , Estrutura Molecular , Fosfolipases A/antagonistas & inibidores , Fosfolipases A1 , Relação Estrutura-AtividadeRESUMO
A new series of anhydrohexitol nucleosides are described. These compounds have a pyrimidine base moiety substituted in the 5-position with a chloro (1b), trifluoromethyl (1c), vinyl (1d), 2-thienyl (1e), ethynyl (1f) or propynyl (1g) substituent. The vinyl, propynyl, and, in particular, the 5-trifluoromethyl analogue showed potent activity against herpes simplex virus (HSV), 1c with a selectivity index of >16000 against HSV-1 and >1000 against HSV-2. Conformational analysis of anhydrohexitol nucleosides using computational methods indicates that these nucleosides occur in an equilibrium between the C1 and 1C form with a DeltaE of 5.9 kJ/mol. When the anhydrohexitol nucleoside is cocrystallized with the HSV-1 thymidine kinase it adopts a 1C conformation, which is opposite to the conformation found for the small molecule alone. The enzyme, apparently, induces a conformational change, and conformational flexibility of an anhydrohexitol nucleoside may be advantageous for recognition by viral enzymes.
Assuntos
Antivirais/síntese química , Arabinonucleosídeos/síntese química , Pirimidinas/síntese química , Antivirais/química , Antivirais/farmacologia , Arabinonucleosídeos/química , Arabinonucleosídeos/farmacologia , Linhagem Celular , Cristalografia por Raios X , Citomegalovirus/efeitos dos fármacos , Herpesvirus Humano 1/efeitos dos fármacos , Herpesvirus Humano 1/enzimologia , Herpesvirus Humano 2/efeitos dos fármacos , Herpesvirus Humano 3/efeitos dos fármacos , Modelos Moleculares , Conformação Molecular , Pirimidinas/química , Pirimidinas/farmacologia , Relação Estrutura-Atividade , Timidina Quinase/químicaRESUMO
The structure-activity relationships of adenosine-3', 5'-bisphosphates as P2Y(1) receptor antagonists have been explored, revealing the potency-enhancing effects of the N(6)-methyl group and the ability to substitute the ribose moiety (Nandanan et al. J. Med. Chem. 1999, 42, 1625-1638). We have introduced constrained carbocyclic rings (to explore the role of sugar puckering), non-glycosyl bonds to the adenine moiety, and a phosphate group shift. The biological activity of each analogue at P2Y(1) receptors was characterized by measuring its capacity to stimulate phospholipase C in turkey erythrocyte membranes (agonist effect) and to inhibit its stimulation elicited by 30 nM 2-methylthioadenosine-5'-diphosphate (antagonist effect). Addition of the N(6)-methyl group in several cases converted pure agonists to antagonists. A carbocyclic N(6)-methyl-2'-deoxyadenosine bisphosphate analogue was a pure P2Y(1) receptor antagonist and equipotent to the ribose analogue (MRS 2179). In the series of ring-constrained methanocarba derivatives where a fused cyclopropane moiety constrained the pseudosugar ring of the nucleoside to either a Northern (N) or Southern (S) conformation, as defined in the pseudorotational cycle, the 6-NH(2) (N)-analogue was a pure agonist of EC(50) 155 nM and 86-fold more potent than the corresponding (S)-isomer. The 2-chloro-N(6)-methyl-(N)-methanocarba analogue was an antagonist of IC(50) 51.6 nM. Thus, the ribose ring (N)-conformation appeared to be favored in recognition at P2Y(1) receptors. A cyclobutyl analogue was an antagonist with IC(50) of 805 nM, while morpholine ring-containing analogues were nearly inactive. Anhydrohexitol ring-modified bisphosphate derivatives displayed micromolar potency as agonists (6-NH(2)) or antagonists (N(6)-methyl). A molecular model of the energy-minimized structures of the potent antagonists suggested that the two phosphate groups may occupy common regions. The (N)- and (S)-methanocarba agonist analogues were docked into the putative binding site of the previously reported P2Y(1) receptor model.
Assuntos
Nucleotídeos de Desoxiadenina/síntese química , Agonistas do Receptor Purinérgico P2 , Antagonistas do Receptor Purinérgico P2 , Ribose/química , Animais , Nucleotídeos de Desoxiadenina/química , Nucleotídeos de Desoxiadenina/farmacologia , Ativação Enzimática , Membrana Eritrocítica/enzimologia , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Técnicas In Vitro , Fosfatos de Inositol/metabolismo , Ligantes , Modelos Moleculares , Receptores Purinérgicos P2Y1 , Relação Estrutura-Atividade , Perus , Fosfolipases Tipo C/metabolismoRESUMO
Following topical application of (125)IVDU, the radiolabeled analogue of bromovinyldeoxyuridine ([E]-5-[2-bromovinyl]-2'-deoxyuridine), as 0.5% or 0.3% eyedrops, to rabbits, (125I)IVDU appeared in the anterior chamber fluid at drug levels well above the minimum concentration (0.01 microgram/mL) required for inhibition of herpes simplex virus type 1 replication. These findings are consistent with the efficacy of 0.5% bromovinyldeoxyuridine eyedrops in the topical treatment of herpes simplex uveitis.
Assuntos
Idoxuridina/análogos & derivados , Ceratite Dendrítica/tratamento farmacológico , Animais , Câmara Anterior/análise , Córnea/análise , Idoxuridina/análise , Idoxuridina/uso terapêutico , Radioisótopos do Iodo , CoelhosRESUMO
Within the polyA:polyT recognition system, cross-pairing between several nucleic acids with a phosphorylated six-membered carbohydrate (mimic) as repeating unit in the backbone structure has been observed. All investigated nucleic acids (except for beta-homo-DNA) hybridize with RNA, leaving RNA as a versatile biopolymer for informational transfer. [reaction: see text]
Assuntos
RNA/química , Conformação de Ácido NucleicoRESUMO
Decomposition products of tylosin A were isolated using open column chromatography and preparative liquid chromatography. Two decomposition products, formed in slightly alkaline medium, were identified as epimers of tylosin A aldol, one of which has been described previously. Another decomposition product was formed on exposure of a tylosin A solution to light. Isomerization of the double bond between C12-C13 takes place, resulting in the formation of the hitherto unreported isotylosin A.
Assuntos
Cromatografia Líquida/métodos , Tilosina/química , Sequência de Carboidratos , Dados de Sequência Molecular , Espectrofotometria UltravioletaRESUMO
Polymyxin B is a peptide antibiotic complex present as sulphate. The components were separated preparatively on a poly(styrene-divinylbenzene) (PLRP-S), 1000 A, 8 microm, 250 x 12.5 mm I.D. stationary phase maintained at 60 degrees C and using 215 nm detection. Elution was carried out with acetonitrile-sodium sulphate solution (0.7%, m/v; pH adjusted to 2.5 with trifluoroacetic acid)-water (18:50:32, v/v) at a flow-rate of 4.0 ml/min. Seven polymyxin B components were isolated and characterized using 1H and 13C NMR. The molecular masses were confirmed by mass spectrometry. The structures of two components were determined for the first time. Polymyxins B5 and B6 were identified as having the same composition as polymyxin B1 except that the fatty acid moiety was nonanoic acid and 3-hydroxy-6-methyloctanoic acid, respectively.
Assuntos
Polimixina B/química , Cromatografia Líquida/métodos , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Peso Molecular , Conformação Proteica , Espectrofotometria UltravioletaRESUMO
The synthesis and 14C-labeling of 3 alpha, 7 alpha, 12 alpha-trihydroxy-27-carboxymethyl-5 beta-cholestan-26-oic acid by two different approaches is described. One of them involves chain elongation of cholic acid via Wittig-Horner condensation of its formylated 24-aldehyde with tetraethyl phosphonoglutarate. The resulting cholestenoate, on deprotection and hydrogenation, affords the unusual C29 bile acid in good yield. An alternative procedure consists in a malonic ester synthesis starting from the formylated 24-alcohol which, after conversion into a mesylate, is reacted with sodium salt of 2-carboethoxy-gamma-butyrolactone. Alkaline hydrolysis, decarboxylation, esterification with diazomethane and selective tosylation of the newly introduced primary hydroxyl function give a C28 precursor, which is easily chain-elongated into a labeled or unlabeled C29 bile acid by reaction with cyanide and hydrolysis. Due to the easy lactonization of some of the C28 intermediates, the latter method provides a better way for introducing a C-29 label than the sequence usually employed for carboxyl labeling of bile acids and consisting in a decarboxylative halogenation of the parent acid followed by substitution of the norhalogenide with [14C]cyanide and hydrolysis. The structure of the synthesized acid or its dimethyl ester is confirmed by 13C nuclear magnetic resonance spectroscopy and mass spectrometry, and is also shown by gas liquid chromatography to be identified with an authentic sample of biosynthetic C29 dioic bile acid extracted from body fluids of Zellweger patients.
Assuntos
Ácidos Cólicos/síntese química , Marcação por Isótopo , Microcorpos/metabolismo , Aldeídos/química , Radioisótopos de Carbono , Ácido Cólico , Ácidos Cólicos/química , Esterificação , Humanos , Hidrólise , Espectroscopia de Ressonância Magnética , Malonatos/química , Mesilatos/química , Estrutura Molecular , Síndrome de Zellweger/metabolismoRESUMO
The glass forming properties of ketoconazole were investigated using differential scanning calorimetry (DSC), by quench cooling liquid ketoconazole from T(m)+10 to 273.1 K, followed by subsequent heating at 5 K/min to T(m)+10 K. It was shown that liquid ketoconazole forms a glass which did not recrystallise following reheating, indicating its stability; T(g) was found to be 317.5+/-0.3 K. However, the presence of a small amount of crystalline ketoconazole was able to convert the amorphous drug back to the crystalline state: the addition of only 4.1% (w/w) of crystalline material converted 77.1% of the glass back to the crystalline state, and this value increased as the amount of added crystals increased. PVP K25 was found to be highly effective in the prevention of such recrystallisation, but only if the amorphous drug was formulated in a solid dispersion, since physical mixing of amorphous ketoconazole with the polymer resulted in recrystallisation of the former compound. Storage of the solid dispersions for 30 days at 298.1 K (both 0 and 52% RH) in the presence or absence of crystals did not result in recrystallisation of the amorphous drug. Solid dispersions formed compatible blends as one single T(g) was observed, which gradually increased with increasing amounts of PVP K25, indicating the anti-plasticising property of the polymer. The values of T(g) followed the Gordon-Taylor equation, indicating no significant deviation from ideality and suggesting the absence of strong and specific drug-polymer interactions, which was further confirmed with 13C NMR and FT-IR. It can be concluded therefore that the physical mechanism of the protective effect is not caused by drug-polymer interactions but due to the polymer anti-plasticising effect, thereby increasing the viscosity of the binary system and decreasing the diffusion of drug molecules necessary to form a lattice.
Assuntos
Antifúngicos/química , Cetoconazol/química , Varredura Diferencial de Calorimetria , Estabilidade de Medicamentos , Espectroscopia de Ressonância Magnética , Excipientes Farmacêuticos , Povidona , Espectroscopia de Infravermelho com Transformada de Fourier , Suspensões , Termogravimetria , Difração de Raios XRESUMO
The isolation and identification of two pinitol alpha-D-galactosides from jojoba meal are described. The products were isolated by a combination of preparative HPLC on silica gel and TLC on amino silica gel and were identified by MS, NMR spectroscopy, and chemical derivatization as 5-O-(alpha-D-galactopyranosyl)-3-O-methyl-D-chiro-inositol or 5-alpha-D-galactopyranosyl-D-pinitol and 2-O-(alpha-D-galactopyranosyl)-3-O-methyl-D-chiro-inositol or 2-alpha-D-galactopyranosyl-D-pinitol. The same preparative HPLC method on silica gel allowed a new simmondsin derivative to be isolated and identified as 4,5-didemethyl-4-O-alpha-D-glucopyranosylsimmondsin mainly by NMR spectroscopy and high-resolution mass spectrometry.
Assuntos
Ração Animal/análise , Cicloexanos , Galactosídeos/isolamento & purificação , Inibidores do Crescimento/isolamento & purificação , Inositol/isolamento & purificação , Ceras/química , Acetonitrilas/análise , Acetonitrilas/isolamento & purificação , Animais , Cromatografia Líquida de Alta Pressão , Galactosídeos/análise , Glucosídeos/análise , Glucosídeos/isolamento & purificação , Inibidores do Crescimento/análise , Inositol/análogos & derivados , Inositol/análise , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Sílica Gel , Dióxido de SilícioRESUMO
Jojoba seed meal shows appetite-suppressing activity due to the presence of simmondsin. This pharmacological activity disappears with treatment of the meal with sodium hydroxide. To elucidate this mechanism of inactivation, the reaction of simmondsin in 1 N NaOH at 20 degrees C was monitored as a function of time. The end products of the reaction as well as intermediates were isolated and identified. The half-life of simmondsin was approximately 60 min with d-glucose and 2-hydroxy-3-methoxyphenylacetonitrile as reaction end products. The reaction mechanism could be elucidated by the isolation of isosimmondsin and a simmondsin lactone derivative. Those compounds were isolated and purified by a combination of column chromatography and HPLC and identified mainly by HRMS and NMR spectroscopy.
Assuntos
Acetonitrilas/química , Cicloexanos , Glucosídeos/química , Magnoliopsida/química , Sementes/química , Hidróxido de Sódio/farmacologia , Cromatografia Líquida de Alta Pressão , Estabilidade de Medicamentos , Glucose/análise , Glucose/química , Meia-Vida , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância Magnética , MetilaçãoRESUMO
The identification of five novel compounds, pseudo-erythromycin A-6,9-hemiketal, 8,9-anhydro-pseudo-erythromycin A-6,9-hemiketal, 8,9-anhydro-pseudo-N-demethylerythromycin A-6,9-hemiketal, 5-O-beta-D-desosaminylerythronolide A and 15-nor-erythromycin C, in mother liquor concentrates of Streptomyces erythraeus is described. The pseudo-erythromycin derivatives are characterized by a 12-membered macrocyclic ring as a result of C13----C11 trans-lactonization. The five compounds have very little antimicrobial activity.
Assuntos
Eritromicina/análogos & derivados , Eritromicina/isolamento & purificação , Streptomyces/análise , Bactérias/efeitos dos fármacos , Meios de Cultura , Eritromicina/farmacologia , Testes de Sensibilidade Microbiana , Relação Estrutura-AtividadeRESUMO
Preparative-scale separation of colistin sulphate bulk sample was carried out on a preparative poly(styrene-divinylbenzene) stationary phase. Isocratic elution with acetonitrile-sodium sulphate solution (0.7% m/v; pH adjusted to 2.5 with TFA) - water (16:50:34, % v/v/v) was carried out at a flow rate of 4.0 ml min(-1). Six colistin components were isolated and characterized using 1H and 13C NMR. The molecular weights were confirmed by mass spectrometry. The structures of 2 components were determined for the first time. Polymyxin E7 was identified as having the same composition as polymyxin E1, except that the fatty acid moiety was 7-methyloctanoic acid. Isoleucine polymyxin E8 was characterized as having the same composition as isoleucine polymyxin E1 with 7-methylnonanoic acid as the fatty acid moiety.