乙酰螺旋霉素

本品系大环内酯类抗菌素螺旋霉素(Spiramycin)的半合成衍生物。     

大环内酯抗菌素的快速(高压)液相色谱

近年来陆续发现,含多种成份的16-元环的大环内酯抗菌素,除碳霉素、螺旋霉素和柱晶白霉素(Leucomycins)(A)外,倘有 maridomycin、YL-704、SF-837和 espinomycin 等。这些抗菌素结构     

乙酰螺旋霉素之临床药理

乙酰螺旋霉素(Acetyl Spiramycin,Ac-SPM)系大环内酯抗菌素螺旋霉素(Spiramycin,SPM)半合成衍生物。1965年由日本合成,国内已于近年来合成并试用于临床.一、理化性质     

16员环大环内酯类抗菌素的化学和生物学研究

当碳霉素和柱晶白霉素的结构研究暂告一段落时,1967～1968年间,新的大环内酯类抗菌素陆续被发现。除了蔷薇霉素(Rosamicin)和铜霉素(Chalcomycin)外,大多数这类抗菌素都是日本研究者发现的。这除了受到柱晶白霉素(Leucomycin)组分结构测定的影响外,还可能是由于柱晶白霉素和螺旋霉素(Spiramycin)在日本临床广泛使用的结果。短期内大量化合物的发现和它们结构的测定推动了抗菌素分离方法的进展。     

螺旋霉素致过敏反应1例报告

螺旋霉素是一大环内酯类抗菌素 ,因其不良反应少 ,临床使用较为广泛 ,近期笔者遇到服用螺旋霉素引起药物性皮疹 1例。现报告如下。患者 ,女 ,3 2岁。因上呼吸道感染、头痛、咽喉痛来院就诊。体检 :咽喉肿 ,双侧扁桃体肿大 ,体温 3 8.1℃。心、肺及腹部无异常。诊断 :急性扁桃体炎。口服螺旋霉素 0 .1g/次 ,tid,并遵医嘱多饮水。服药次日患者眼部出现点状红疹 ,手指压下即褪色 ,伴有搔痒 ,3天后皮疹减少。2个月后患者因扁桃体炎又自服螺旋霉素 ,面部出现红疹 ,小米粒大 ,大小均匀 ,边界清楚 ,逐渐波及到前胸上肢 ,压不褪色 ,奇痒 ,又来院就诊…     

黄豆饼粉对螺旋霉素合成的影响

黄豆饼粉是许多抗生素发酵培养基中较好的氮源,而螺旋霉菌如采用黄豆饼粉为主要氮源与生产用的鱼粉为主要氮源的培养基比较,则螺旋霉素合成有很显著的差异。为要提供工业生产抗生素的有用资料,我们对此作了进一步研究。 材料和方法 (-)菌种:我国土壤中分离到的螺旋霉素产生菌 Streptomyces spiramyceticusA.SP编号为——799-1941. (二)培养基 1.孢子斜面培养基:麦敖6％,琼脂2％ 2.种子培养基:黄豆饼粉25％,淀粉4％,NaCl0.4％,CaCO_30.5％,自来水,自然pH,500毫升三角瓶装量100毫升。 3.发酵培养基:     

提高基因工程菌产二素链霉菌311—10产生酰化螺旋霉素的研究

基因工程菌产二素链霉菌(Str.ambofaciens)311-10是一株含麦迪霉素4″—羟基酰化酶基因的螺旋霉素产生菌,可发酵直接产生酰化螺旋霉素。经质谱鉴定,所产生的酰化产物为4″—异戊酰、4″—丁酰及4″—丙酰螺旋霉素,并含有50～60％的螺旋霉素组份。为了提高酰化螺旋霉素的含量,研究了17种氨基酸及有关化合物对产二素链霉菌311-10菌株直接产生酰化螺旋霉素的影响。结果显示,在发酵过程中添加一定量α—羟基异丁酸或L—异亮氨酸,酰化螺旋霉素的的百分含量可从对照组的45％分别提高到73％和67％。如同时加入α-羟基异丁酸和L-异亮氨酸.则酰化螺旋霉素的含量可增至88％左右。研究表明含麦迪霉素4″—羟基酰化酶基因的螺旋霉素产生菌能利用L—异亮氨酸或α—羟基异丁酸作为酰基的供体。从不同发酵时间加入试验表明合适的添加时间为发酵前期24小时左右。提示L—异亮氨酸或α—羟基异丁酸不只是提供酰化螺旋霉素4”—酰化酶酰基的供体,可能对4″—酰化酶基因的转录起诱导作用。 L—精氨酸能使酰化螺旋霉素组份下降至14％左右,并能抵消α-羟基异丁酸或L-异亮氨酸对酰化螺旋霉素的促进作用。 研究提示控制基因工程菌发酵条件,可以有效地获得所需基因工程产物。     

中心组合设计优化必特螺旋霉素合成培养基

将克隆自卡波霉素产生菌的4＂-O-异戊酰基转移酶基因整合到螺旋霉素产生菌Streptomyces spiratnvceticus F-21的染色体上，构建成一株稳定的生物工程菌WSJ-1-195，它产生的一组以4＂-O-异戊酰螺旋霉素为主要成分的多组分基因工程新型抗生素命名为必特螺旋霉素。针对目前没有适合必特螺旋霉素产生菌的合成培养基，所以本文顺序通过部分因子析因设计法、最速上升实验、中心组合实验，并利用统计学软件SAS V8对实验数据进行分析，确定了必特螺旋霉素合成培养基的组成，为以后对必特螺旋霉素生理生化特性的研究提供基础。经过优化后必特螺旋霉素的发酵效价从173μg／ml提高到1880μg／ml。     

溶剂浸渍树脂的制备及其在提取螺旋霉素上的应用

溶剂浸渍树脂是一种正在发展的分离介质。本文首次研究了醋酸戊酯浸渍树脂吸附分离螺旋霉素的工艺条件。浸渍树脂对螺旋霉素的吸附量不低于大孔吸附剂,用pH为4含15％乙醇的醋酸缓冲液作为解吸剂。解吸率达到95％以上。直接合成制得的浸渍树脂对螺旋霉素的吸附量亦较高。研究表明,浸渍树脂在分离过程中溶剂的萃取作用和吸附剂的表面吸附作用同时发生。测定了浸渍树脂的吸附等温线和吸附、解吸条件,提出了从发酵液中提取螺旋霉素的工艺过程。     

生技霉素E(4"-异戊酰螺旋霉素Ⅰ)的分离和结构鉴定

为获得高产的4"异戊酰螺旋霉素,将4"异戊酰基转移酶基因整合入螺旋霉素产生菌S.spiramyceticusF21染色体,构建成功了一株稳定的生物工程菌WSJ1,继从该菌株代谢产物中分离获得生技霉素A1(4"异戊酰螺旋霉素)、生技霉素B1(4"异戊酰螺旋霉素),又分离获得一主要组分生技霉素E。经UV、FTIR、HRSIMS、1HNMR、13CNMR、1H1HCOSY等光谱分析和与生技霉素A1及相关文献的光谱数据比较,确定生技霉素E为本菌株目标化合物之一——4"异戊酰螺旋霉素,高效液相分析表明,该生物工程菌代谢产物中4"异戊酰螺旋霉素含量超过55%。     

两株特境链霉菌产抗MRSA次级代谢产物结构类型的快速鉴定

目的对链霉菌ZZ027和ZZ021所产抗耐甲氧西林金黄色葡萄球菌(MRSA)次级代谢产物的结构类型进行快速鉴定。方法用2A培养基对两链霉菌菌株进行发酵;再采用HPLC微分离技术和琼脂扩散法对抗MRSA色谱峰进行定位,然后采用TLC对发酵液中抗MRSA次级代谢产物进行分离与制备,所得抗MRSA组分经薄层显色、IR和13C-NMR谱测试分析,鉴定其结构类型。结果链霉菌ZZ027和ZZ021抗MRSA组分的薄层显色、IR和13C-NMR谱特征均与大环内酯类化合物的相关表征一致。结论链霉菌ZZ027和ZZ021所产抗MRSA次级代谢产物为大环内酯类化合物。     

Microbial transformation of aniline to acetaminophen

In order to obtain acetaminophen, a popular analgesic-antipyretic, through microbial p-hydroxylation and N-acetylation of aniline, various fungi and bacteria were screened. Among them,Streptomyces species were chosen for strain improvement by the use of interspecific protoplast fusion technique. Two interspecific fused strains were developed betweenS. rimosus (N-cetylation function) andS. aureofaciens (p-hydroxylation function) and also betweenS. lividans andS. globisporus. For efficient protoplast fusion and cell wall regeneration, various conditions were examined. In a typical experiment of mixedS. rimosus (pro⁻ his⁻) andS. aureofaciens (ilv⁻) protoplasts with 40% (w/v) polyethylene glycol 3350 (PEG) for 3 min gave 8.3×10⁻⁷ of fusion frequency. Treatment of mixedS. lividans (pant⁻) andS. globisporus (leu⁻) protoplasts with 50% (w/v) PEG for 3 min at 30°C gave 1.2×10⁻⁶ of frequency. Among the fused strains, up to 40–50% increase in p-hydroxylation power was observed. To investigate the possibility of plasmid involvement in p-hydroxylation of acetanilide, plasmid curing was attempted. We found that cells treated with acriflavine (at the frequency of 100%) and cells regenerated from protoplsts ofS. aureofaciens (2% frequency) lost their p-hydroxylation function.     

Spiramycin in triple therapy of Helicobacter pylori-associated peptic ulcer disease. An open pilot study with 12-month follow-up

Background: Spiramycin (Rovamycin) is a well-established macrolide antibiotic with good anti-Helicobacter pylori activity in vitro. It is acid-stable and found in high concentration in various body fluids and cells after oral administration. Its anti-H. pylori activity in vivo has not yet been tested. Methods: Twenty-five consecutive patients with endoscopically verified peptic ulcer and a positive biopsy urease test were given spiramycin tables 1.5 MIU instead of oxytetracycline 500 mg q.d.s. in our triple therapy regimen with bismuth subnitrate suspension 10 mL (150 mg bismuth subnitrate) q.d.s. and metronidazole 400 mg t.d.s. for 10 days. Bismuth was taken between meals and spiramycin and metronidazole with meals. Re-endoscopy and ¹⁴C-urea breath test were performed 4 weeks after completion of therapy. Those who were H. pylori negative according to the breath test returned for 1-year follow-up. Results: Per protocol analysis at 4 weeks showed that 21 out of 23 patients were H. pylori negative and had healed ulcers. These 21 patients were persistingly H. pylori negative and had no ulcers at 1-year follow-up. H. pylori eradication and ulcer healing rates were thus 91.3%; 95 % confidence interval from 72.0% to 98.9%. Side-effects limiting daily activity were significantly less frequent than we have experienced previously using oxytetracycline in triple therapy. Conclusions: Spiramycin appears to be an alternative to tetracycline in the triple therapy of H. pylori infection. Further studies to position spiramycin as an anti-H. pylori drug are warranted.     

A new streptothricin family antibiotic producingStreptomyces spp. SNUS 8810-111; Characterization of the producing organisms,fermentation, isolation,and structure elucidation of antibioitics

A new streptothricin family antibiotic producingStreptomyces spp. SNUS 8810-111 was isolated from a soil sample. Study of its morphological and physiological characters indicated that the antibiotic producing organism was aStreptomyces spp. Taxonomical studies suggested that the organism might belong to the genusStreptomyces gougeroti. The organism produced antibiotics most in calcium carbonate-tryptic soy broth. The active principles were recovered from the broth with a cation exchange resin and eluted from the resin with HCl. Cellulose column chromatography gave two active principles.¹H-¹H Homo-COSY study on the first compound revealed four structural components. Total hydrolysis of the antibiotic with HCl allowed isolation of β-lysine. From these data the antibiotic was found to be streptothricin D. The other compound showed one additional signal in the¹H NMR and the¹³C NMR spectra. The signal was from a methyl group attached to a nitrogen atom. Comparison of the NMR signals with those of streptothricin D suggested that the compound wasN-methyl-streptothricin D which was a new compound in the family of streptothricin antibiotics.     

Activity of benzalkonium chloride and chlorhexidine diacetate against wild-type and envelope mutants of Escherichia coli and Pseudomonas aeruginosa

Whole cells of wild-type (DCO) and envelope mutant (DC2) strains of Escherichia coli took up approximately equal amounts of the cationic surfactant, benzalkonium chloride, although the mutant was considerably more sensitive to this bactericide. Lower concentrations of benzalkonium were needed to induce K⁺ leakage from the mutant than from the parent cells. DCO and DC2 showed the same order of sensitivity to chlorhexidine diacetate (minimum inhibitory concentrations against single cell inocula, 1.5 and 0.4 $\text{μg}\text{ml}$, respectively) and took up approximately equal amounts of this antiseptic. K⁺ leakage was much greater from DC2 exposed to benzalkonium and slightly higher from chlorhexidine-treated DC2 than from drugtreated DCO. Similar studies with wild type (799) and envelope mutant ($\text{799}\text{61}$) strains of Pseudomonas aeruginosa showed that they took up similar amounts of benzalkonium or of chlorhexidine from solution. However, somewhat greater leakage of K⁺ occurred from $\text{799}\text{61}$ than from 799. Lysozyme-EDTA spheroplasts of a wild-type strain and its envelope mutant were equally sensitive to an antibacterial agent.     

Production, isolation and biological activity of nargenicin from Nocardia sp. CS682

Culture broth of an actinomycete isolate, Nocardia sp. CS682 showed specifically higher antibacterial activity against methicilin resistant Staphylococcus aureus (MRSA). Purified substance from the organism, CS-682, which is active against MRSA and Micrococcus leuteus, is a C₂₈H₃₇NO₈ (M+H⁺, observed: 516.83) and identified as an unusual macrolide antibiotic, nargenicin. The chemical structure of CS-682 was identified by FT-IR, ¹H-NMR, ¹³C-NMR, and (¹H-¹H and ¹H-¹³H) COSY. The anti-MRSA activity of CS-682 was stronger than that of oxacillin, vancomycin, monensin, erythromycin, and spiramycin. Phylogenetic analysis showed that strain CS682 is closely related to Nocardia tenerifensis DSM 44704^T (98.7% sequence similarity), followed by N. brasiliensis ATCC 19296^T (98.4% sequence similarity). The ability of Nocardia sp. CS682 to produce nargenicin was unique.     

Ethanol inhibits the N-methyl-D-aspartate (NMDA)-induced attenuation of the NMDA-evoked noradrenaline release in the rat brain cortex: interaction with NMDA-induced desensitization

Summary The influence of ethanol, AP5 (DL-2-amino-5-phosphonopentanoic acid) and dizocilpine (MK-801) ((+)-5-methyl-10, 11-dihydro-5H-dibenzo (a, b)-cyclohept-5,10-imine hydrogen maleate) on the NMDA-induced attenuation of the NMDA-evoked noradrenaline release was examined in rat brain cortex slices preincubated with ³H-noradrenaline. The slices were superfused with Mg²⁺-free Krebs-Henseleit solution. Tritium overflow (corresponding to ³H-noradrenaline release) was stimulated by 300 mol/l NMDA for 2 min.Presence of 10–100 mol/l NMDA from 20 to 2 min before stimulation concentration-dependently inhibited the NMDA (300 mol/l)-evoked ³H overflow, suggesting an agonist-induced desensitization attributable to the modification of events at the NMDA receptor itself and/or distal to this receptor system. The desensitizing effect of 100 mol/l NMDA was almost complete and was not diminished when the time of preexposure was decreased to 10 min, and when NMDA was removed from the superfusion fluid for up to 5 min before the stimulus; however, the densitizing effect was reduced after further decrease in the duration of preexposure to NMDA or further prolongation of the interval between preexposure and stimulation. Ethanol inhibited the NMDA-induced ³H overflow (IC₅₀ 45 mmol/l); this effect was almost abolished when ethanol was omitted from the superfusion fluid from 2 min before stimulation onward. Ethanol, when simultaneously present with 100 mol/l NMDA in the superfusion fluid from 20 to 2 min before the NMDA stimulus, concentration-dependently (IC₅₀ 112 mol/l) decreased the inhibitory effect of NMDA. The effect of ethanol was identical to that of the competitive NMDA receptor antagonist AP5 both with respect to the rapid reversibility of the inhibition of NMDA induced ³H overflow (i.e. within 2 min) and to the decrease of the desensitizing effect of NMDA. The pattern of effects obtained with dizocilpine, an inhibitor of the ion channel coupled to the NMDA receptor, was different from that obtained with ethanol and AP5: the inhibitory effect of dizocilpine on the NMDA-evoked ³H overflow was only partly reversible within 8 min after withdrawal, and the inhibitory effect of NMDA on NMDA-evoked ³H overflow was not decreased by dizocilpine, but was more pronounced than with either NMDA or dizocilpine alone.It is concluded that ethanol mimics AP5 in that in inhibits the NMDA induced desensitization, possibly by preventing the NMDA receptor from being activated by NMDA applied during the desensitization period. In contrast, dizocilpine produces no such effect. The potency of ethanol in inhibiting the NMDA-induced desensitization was about 3 times less than in inhibiting NMDA-evoked noradrenaline release. These results are compatible with the suggestion that the NMDA recognition site itself may be one of the sites of action of ethanol and that ethanol does not modify the function of the NMDA receptor system by a machanism comparable to that of dizocilpine. Send offprint requests to M. Göthert at the above address     

Is the Nephrotoxicity of (R)-3-Chlorolactate in the Rat Caused by 3-Chloropyruvate?

1. When (R, S)-[3-³⁶Cl]chlorolactate was administered to male rats, two radioactive constituents were excreted in the urine. These were identified as ³⁶Cl^? and [3-³⁶Cl]chlorolactate which was subsequently shown to be essentially the (S)-isomer.

2. Analysis of the urinary oxalate content from rats receiving either (R)- or (S)-3-chlorolactate revealed that elevated levels were produced by the (R)-isomer whereas normal levels followed the administration of the (S)-isomer.

3. Treatment of (R, S)-3-chlorolactate with a modified Fenton' oxidizing system produced oxalate and an intermediate which was identified as 3-chloropyruvate.

4. 3-Chloropyruvate is a potent nephrotoxin in the rat producing a brief phase of diuresis when administered, increasing the urinary excretion of oxalate and inhibiting the oxidative metabolic capability of rat kidney tubules and rat kidney mitochondria in vitro.

5. Both (R)-3-chlorolactate and 3-chloropyruvate were shown to be inhibitors of the commercially-available pyruvate dehydrogenase complex.

6. 3-Chloropyruvate inhibits kidney mitochondrial metabolism possibly at the pyruvate dehydrogenase complex level and appears to be a metabolite of (R)- but not (S)-3-chlorolactate.     

Aspergillus flavus Growth Inhibition and Aflatoxin B1 Decontamination by Streptomyces Isolates and Their Metabolites

Aflatoxin B₁ is a potent carcinogen produced by Aspergillus flavus, mainly during grain storage. As pre-harvest methods are insufficient to avoid mycotoxin presence during storage, diverse curative techniques are being investigated for the inhibition of fungal growth and aflatoxin detoxification. Streptomyces spp. represent an alternative as they are a promising source of detoxifying enzymes. Fifty-nine Streptomyces isolates and a Streptomyces griseoviridis strain from the commercial product Mycostop^®, evaluated against Penicillium verrucosum and ochratoxin A during previous work, were screened for their ability to inhibit Aspergillus flavus growth and decrease the aflatoxin amount. The activities of bacterial cells and cell-free extracts (CFEs) from liquid cultures were also evaluated. Fifty-eight isolates were able to inhibit fungal growth during dual culture assays, with a maximal reduction going down to 13% of the control. Aflatoxin-specific production was decreased by all isolates to at least 54% of the control. CFEs were less effective in decreasing fungal growth (down to 40% and 55% for unheated and heated CFEs, respectively) and aflatoxin-specific production, with a few CFEs causing an overproduction of mycotoxins. Nearly all Streptomyces isolates were able to degrade AFB₁ when growing in solid and liquid media. A total degradation of AFB₁ was achieved by Mycostop^® on solid medium, as well as an almost complete degradation by IX20 in liquid medium (6% of the control). CFE maximal degradation went down to 37% of the control for isolate IX09. The search for degradation by-products indicated the presence of a few unknown molecules. The evaluation of residual toxicity of the tested isolates by the SOS chromotest indicated a detoxification of at least 68% of AFB₁’s genotoxicity.