New melanin biosynthesis inhibitors and their structural similarities

Relationships between their activities as blast control agents, and their abilities to inhibit mycelial melanisation on a nutrient agar, are described for 103 substituted benzothiazol-2(3H)-ones, benzoxazol-2(3H)-ones, indolin-2-ones, quinolin-2(1H)-ones, 1,2,3,4-tetrahydroquinolin-2-ones, benzo-1,4-thiazin-3(2H)-ones and benz-1,4-oxazin-3(2H)-ones, and some corresponding thiones. Several compounds in the respective series had a high protective activity and an antimelanisation activity against the blast fungus Pyricularia oryzae; furthermore, there was a good correlation in both of these activities, indicating that these compounds belong to the group of melanin biosynthesis inhibitors. Structural similarities of these compounds can be identified as follows: (a) having a benzo-bicyclic ring system; (b) containing a nitrogen atom in one ring at a position alpha to the benzene ring system; and (c) substitution, at the ring nitrogen atom, at the peri position in the aromatic ring relative to the nitrogen atom, and at the position alpha to the nitrogen atom in the ring system, with a double bond such as in carbonyl and thiocarbonyl groups. Among the compounds that have been proposed as melanin biosynthesis inhibitors, the chemical structures of tricyclazole, pyroquilon, 4,5-dihydro-4-methyltetrazolo[1,5-a]quinazolin-5-one (PP-389), [1,2,4]-triazolo[4,3-a]quinoline and 1-methylquinolin-2(1H)-one exhibit the structural similarities described above; however, 4,5,6,7-tetrachlorophthalide, 2,3,4,5,6-pentachlorobenzyl alcohol and N-substituted-2,3,4,5-tetrachloro-6-(hydroxymethyl)benzamides do not have such similarities in their chemical structures.     

The effect of three fungicides,specific for the control of rice blast disease,on the growth and melanin biosynthesis by Pyricularia oryzae cav.

Tricyclazole (EL 291), 4,5-dihydro-4-methyltetrazolo [1,5-a] quinazolin-5-one (PP 389), and pyroquilon (CGA 49104) were studied to determine the effects on growth and melanin biosynthesis by Pyricularia oryzae in vitro. The three fungicides were essentially devoid of toxicity to P. oryzae at concentrations up to 50 ug ml^?1 but each selectively inhibited melanin biosynthesis at much lower concentrations. Inhibition of melanin biosynthesis resulted in the accumulation of 2-hydroxyjuglone and flaviolin. The study indicated that the three compounds act by a similar mechanism in P. oryzae..     

Fungitoxicity of 6H-pyrazolo[3,4-c][l,2,5]thiadiazine-2,2-dioxides

A base-promoted cyclisation of 4-nitroso-5-benzylsulphonamidopyrazoles (VII) afforded 6 H -pyrazolo[3,4-c][1,2,5]thiadiazine-2,2-dioxides (VIII). They were tested in vitro for antifungal activity against a series of phytopathogenic fungi of different taxonomic classes. Five of the compounds had noteworthy activity; in particular 6H-3-phenyl-5-methyl-7–(3,4-dichlorophenyl) pyrazolo[3,4-c][1,2,5]thiadiazine-2,2-dioxide (VIIIc) at the concentration of 200 mg litre^?1 completely inhibited the growth of Pythium ultimum, Corticium solani and Sclerotinia minor.     

异香豆素噁二唑类化合物的合成及除草活性

为了寻找具有较高除草活性的异香豆素类化合物,设计合成了16个新型异香豆素噁二唑类化合物4a～4p,其结构均经核磁共振氢谱、碳谱及高分辨质谱确证。初步除草活性测定结果表明,部分目标化合物在500 mg/L下对马唐Digitaria sanguinalis L.和反枝苋Amaranthusretroflexus L.具有一定的除草活性,其中化合物4j（4-[5-（4-氯-苯基）-[1,2,4]二唑-3-基]-3-己基-6,7-二甲氧基异香豆素）的活性最好,对马唐和反枝苋的致死率分别为64%和62%。     

Bicyclophosphorothionate antagonists exhibiting selectivity for housefly GABA receptors

2,6,7-Trioxa-1-phosphabicyclo[2.2.2]octane 1-sulfides (bicyclophosphorothionates) with various C_1–4 alkyl groups at the 3- and 4-positions were synthesized and tested for their ability to compete with [³H]4′-ethynyl-4-n-propylbicycloorthobenzoate (EBOB), a non-competitive antagonist of γ-aminobutyric acid (GABA) receptors, for specific binding to rat-brain and housefly-head membranes, and for their insecticidal activity against houseflies. Among the 3,4-substituted analogues, 20 compounds were selectively active for housefly GABA receptors versus rat GABA receptors. The 3-alkyl groups of C₃ length and the 4-alkyl groups of C₄ length were tolerated in housefly receptors, whereas such bulky substituents were deleterious in rat receptors. The 4-isobutyl-3-isopropyl analogue was the most potent in housefly receptors (IC₅₀ = 45.2 nM ), and tert-butylbicyclophosphorothionate (TBPS), with the 4-tert-butyl group and no 3-substituent, was the most potent in rat receptors (IC₅₀ = 62.2 nM ). Their receptor selectivities (rat IC₅₀/housefly IC₅₀) were 52 and 0.038, respectively. The insecticidal activity (LD₅₀) of 20 active analogues was well correlated with their potency (IC₅₀) in inhibiting [³H]EBOB binding to housefly-head membranes (r = 0.93). The results obtained in the present study indicate that the introduction of appropriate alkyl groups into the 3- and 4-positions of bicyclophosphorothionate leads to non-competitive antagonists with increased affinity and selectivity for housefly ionotropic GABA receptors versus rat GABA_A receptors. © 1999 Society of Chemical Industry     

Inhibition of chitin biosynthesis by buprofezin analogs in relation to their activity controlling Nilaparvata lugens Stål

Buprofezin (Applaud, 2-tert-butylimino-3-isopropyl-5-phenyl-3,4,5,6-tetrahydro-2H-1,3,5-thiadiazin-4-one) strongly inhibited the [³H]chitin synthesis from N-acetyl-d-[1-³H]glucosamine in the brown rice planthopper, Nilaparvata lugens Stål. No inhibition was observed for [³H]-labeled protein biosynthesis from [5-³H]glucose or l-[3,5-³H]tyrosine but [³H]-labeled nucleic acid synthesis from [5-³H]glucose was weakly reduced by buprofezin. The lethal activity of buprofezin analogs related well to their inhibitory potency against chitin biosynthesis in N. lugens nymphs.     

Fungitoxicity of 5-aminoisoxazole-4-thiocyanate derivatives

3-Methyl-5-(acyl/alkyl)aminoisoxazole-4-thiocyanates were prepared by rhodanation of N-acyl/alkyl-3-methyl-5-isoxazolamines. These products were tested for antifungal activity against a series of phytopathogenic fungi of different taxonomic classes. Some of the compounds showed interesting in-vitro activity. The more active compound in the in-vitro test displayed a moderate preventive activity against Plasmopara viticola, Alternaria solani and Pyricularia recondita.     

Independent and joint biological activity of isomeric forms of synthetic pyrethroids

Various isomeric mixtures of pyrethroids were examined in topical application tests against houseflies, Musca domestica. On the basis of the activities of the separate isomers of 5-benzyl-3-furylmethyl (±)-cis,trans-chrysanthemate, it was shown that when combined in pairs to give the (±)-trans or (±)-cis or (+)-cis,trans mixtures the observed mortalities did not differ from those expected by simple additive action calculated by the harmonic mean. In contrast the (±)-cis,trans mixture showed considerable antagonism with a mortality only 60% of that expected. Similar evaluations using the separate and combined isomers of bioallethrin [(R,S)-3-allyl-2-methyl-4-oxocyclopent-2-enyl (allethronyl) ( + )-trans-[(1R,3R)-chrysanthemate] and the corresponding (+)-cis-(1R,3S)-chrysanthemate indicate antagonism calculated to be correlated with the content of the (R)-isomer of the alcoholic moiety. Hence the activity of the most active isomer of the “allethrin” series, (S)-3-allyl-2-methyl-4-oxocyclopent-2-enyl ( + )-trans-(1R,3R)-chrysanthemate, (S)-bioallethrin, is not fully realised unless it is present in pure form and a substantial part of the value of bioresmethrin (5-benzyl-3-furylmethyl ( + )-trans-chrysanthemate] as a killing agent is lost when the racemic form is used. In racemic mixtures there is mutual antagonism between pairs of isomers so that considerable masking of activity occurs.     

嘧啶联吡唑甲酰胺类化合物的合成及除草活性

设计合成了一系列结构新颖的嘧啶联吡唑甲酰胺类化合物5a~5o,其结构均经过1H NM R和MS分析确证。初步生物活性测试结果表明:在有效成分150 g/hm2剂量下苗后茎叶喷雾处理时,化合物(R)-N-[1-(4-氯苯基)乙基]-3-二氟甲基-1-(4,6-二甲氧基嘧啶-2-基)-1H-吡唑-4-甲酰胺(5c)、N-[1-(4-氯苯基)乙基]-1-(4,6-二甲氧基嘧啶-2-基)-N-甲基-3-三氟甲基-1H-吡唑-4-甲酰胺(5i)和N-[1-(4-氯苯基)乙基]-1-(4,6-二甲氧基嘧啶-2-基)-3-三氟甲基-1H-吡唑-4-甲酰胺(5k)对繁缕Stellaria media的抑制率高达90%以上;而同样剂量下苗前土壤喷雾处理时,化合物N-[1-(4-氯苯基)乙基]-3-二氟甲基-1-(4,6-二甲氧基嘧啶-2-基)-1H-吡唑-4-甲酰胺(5b)和5c对繁缕的抑制率达100%。该类结构化合物有望作为除草先导化合物进行开发。     

Heterocyclic Insecticides Acting at the GABA-Gated Chloride Channel: 5-Alkyl-2-arylpyrimidines and -1,3-thiazines

5-tert-Butyl-2-(4-ethynylphenyl)pyrimidine and the corresponding 2,5-disubstituted-4H-1,3-thiazine block the GABA-gated chloride channel at c.20and c.200 nm , respectively, measured as 50% inhibition of the binding of 1-(4-ethynylphenyl)-4-[³H]propyl-2,6,7-trioxabicyclo[2.2.2]octane (4′-ethynyl-4-n-[³H]propylbicycloorthobenzoate; [³H]EBOB) in house fly and mouse brain membranes, and they are also toxic to topically-treated flies with LD₅₀ values of 6–27 μg g⁻¹ alone and 2–6 μg g⁻¹ with piperonyl butoxide (PB) as synergist. In the pyrimidine series, the general pattern of effectiveness of substituents in the 5-position is tert-butyl>isopropyl≈cyclohexyl≈cyclopropyl>methyl, phenyl and 3- and 4-fluorophenyl, and in the 2-position is 4-ethynylphenyl≪4-bromophenyl. These planar pyrimidines and nearly-planar 4H-1,3-thiazines with 2-ethynylphenyl or 2-bromophenyl and 5-tert-butyl or 5-isopropyl substituents are more effective than the corresponding 6H-1,3-thiazine, 6-oxo-1,3-thiazines and 4,6-dioxo-1,3-thiazine examined, but they are less active than the analogous conformationally flexible trans-1,3-dioxanes and -1,3-dithianes. The heterocyclic moiety confers a region of high electron density and positions the 2- and 5-substituents in a linear or parallel relationship for optimal affinity at the receptor. Two observations indicate that the new pyrimidines and thiazines probably act as chloride channel blockers. First, the poisoning signs are identical to those of EBOB in both mice and house flies. Second, each of the pyrimidines, thiazines and dioxanes falls on the same correlation line for inhibition of [³H]EBOB binding and toxicity to house flies (with PB) as that obtained earlier for EBOB analogs, dithianes and polychlorocycloalkanes, suggesting that they all act at the same or closely coupled binding sites in the GABA-gated chloride channel.     

杨梅叶抑菌活性成分初步分离与鉴定

用石油醚、氯仿、乙酸乙酯和正丁醇分别对杨梅叶甲醇提取物进行了萃取,得不同萃取相。测定了其对番茄灰霉病菌Botrytis cinerea、番茄早疫病菌Alternaria solani、水稻立枯丝核Rhizoctonia solani、小麦禾谷镰刀菌Fusarium graminearum、 黄瓜炭疽病菌Colletotrichum lagenarium、玉米大斑病菌Setosphaeria turcica等植物病原菌的抑菌活性。结果表明,乙酸乙酯相和正丁醇相的抑菌活性较好。对乙酸乙酯相进一步分离得到7个化合物,经电喷雾质谱(ESI-MS)、核磁共振氢谱(1H NMR)和碳谱(13C NMR)鉴定其分别为已知化合物山楂酸( 1 )、3-O-反-对-香豆酰基马斯里酸( 2 )、3-O-没食子酰二氢杨梅素( 3 )、斛皮素-7-O-鼠李糖甙( 4 )、杨梅素3-O-鼠李糖甙( 5 )、杨梅素-3-O-鼠李糖(3-O-没食子酰)甙( 6 )和杨梅素-3-O-鼠李糖(2-O-没食子酰)甙( 7 )。孢子萌发法测定结果表明,化合物 3~6 对番茄灰霉病菌和玉米大斑病菌的孢子萌发具有较好的抑制作用,EC50值分别在3.44~4.59 mg/L和8.85~11.6 mg/L之间。     

The fungicidal activity of substituted 1,4-naphthoquinones. Part III: Amino,anilino and acylamino derivatives

A variety of 2-amino-, 2-anilino- and 2-acylamino-1, 4-naphthoquinones, with hydrogen, chlorine, alkoxy, phenoxy, methylthio or phenylthio as the 3-substituent, have been synthesised and assayed against Botrytis cinerea, Cladosporium fulvum and Venturia inaequalis. With few exceptions, only the 2-acylamino compounds possessed appreciable fungicidal activity: this was of a high order in the cases of 2-prop-ionamido-, 2-N-methylacetamido-3-methylthio-, and 3-methoxy-2-N-methylacetamido-1, 4-naphthoquinones.     

Effect of abamectin on electrophysiological activity in the ventral nerve cord of Periplaneta americana L.: Correlation with symptoms of poisoning and levels of toxicant in vivo

Twenty-four hours after injection of adult male Periplaneta americana with 36 ng [5-³H]abamectin, when symptoms of poisoning were apparent in most individuals, similar levels of radioactivity were found in tissue samples from the ventral nerve cord, metathoracic muscle, fat body and testis of randomly-selected insects. When insects injected with [5-³H]abamectin (36 ng) were separated after 24 h into three groups showing varying symptoms, the level of radioactivity in the ventral nerve cord was found to be significantly greater in partially-paralysed and paralysed groups (4- and 7-fold respectively) than in a treated but non- paralysed group. The mean levels of abamectin in the nerve cords of unaffected, ‘partially-paralysed’ and ‘paralysed’ insects were estimated to be 4, 23 and 37 nM respectively. Extracellular studies of the in-vitro action of abamectin (10 nM and 1 μM) on the spontaneous activity of a picrotoxin-sensitive, partially desheathed, ventral nerve cord preparation of adult male P. americana found the mean response time to be relatively slow (77 and 38 min respectively). In sheathed nerve cords, the mean response time to abamectin (1 μM) was 117 min. In all cases, treatment with abamectin increased the stimulus voltage required to evoke a response. Spontaneous activity was found to be progressively reduced in ventral nerve cord preparations from ‘partially-paralysed’ and ‘paralysed’ insects compared with ‘non-paralysed’ and untreated insects following dissection 24 h after injection of P. americana with unlabelled or labelled abamectin (36 ng), while the stimulus required to evoke activity in the ventral nerve cord showed the reverse trend. These differences in electrophysiological activity could be correlated directly with varying levels of abamectin in the nerve cords of ‘paralysed’ and ‘partially-paralysed’ insects.     

Synthetic pyrethroids containing a C–C triple bond

The three commercial synthetic pyrethroids containing a carbon–carbon triple bond, α-ethynyl-2-methylpent-2-enyl (1R)-trans-chrysanthemate, (S)-2-methyl-4-oxo-3-(2-propynyl)cyclopent-2-enyl (1R)-trans,cis-chrysanthemate and [2,5-dioxo-3-(2-propynyl)-1-imidazolidinyl]methyl (1R)-trans-chrysanthemate are reviewed with emphasis on their inventive histories. Their chemistry and efficacy are described briefly. The relationship between stereochemistry and the biological activity is also discussed. © 1998 SCI.     

Phytotoxic activity of norbornane and nortricyclene lactones

A series of bromo-, hydroxy- and keto-derivatives of norbornane-6-hydroxy-2-carboxylic acid, y-lactone and some related compounds have been prepared and examined for phytotoxic activity. The 3-exo-propionyl, and 3-exo-5-exo-dibromo substituted compounds have been found to have the greatest activity.     

Synthesis and Anthelmintic Activity of 7-Hydroxy- 5-oxo-5H-thieno[3,2-b]pyran-6-carboxanilides and -6-thiocarboxanilides

7-Hydroxy-5-oxo-5H-thieno[3,2-b]pyran-6-carboxanilides and -6-thio-carboxanilides represent a novel series of anthelmintic compounds, with broad-spectrum activity against important parasitic nematodes in sheep and dogs. In particular, an improved efficacy against Trichostrongylus colubriformis in sheep over the related 3-carbamoyl-4-hydroxycoumarins has been noted. New synthetic routes to the key intermediate, 7-hydroxythieno[3,2-b]pyran-5-one, have been developed.     

棉铃虫四个丝氨酸蛋白酶抑制剂基因的克隆与表达分析

为明确棉铃虫Helicoverpa armigera丝氨酸蛋白酶抑制剂（serine protease inhibitor,serpin）的种类及其表达特性,利用PCR技术克隆棉铃虫的serpin基因,使用生物信息学软件预测其结构并进行系统进化分析,采用实时荧光定量PCR（real time quantitative PCR,RT-qPCR）技术比较serpin基因在棉铃虫不同发育阶段和组织中的表达量及取食Cry1Ac后其表达量的变化。结果表明,共获得serpin-a、serpin-b、serpin-c、serpin-e四个棉铃虫serpin基因,全长为1 119~1 254 bp,编码373~418个氨基酸,均包含一段具有反应中心环的保守结构域,且与斜纹夜蛾Spodoptera litura、草地贪夜蛾S.frugiperda等鳞翅目昆虫serpin的同源性较高。serpin-a和serpin-e在棉铃虫4龄幼虫期的表达量最高,serpin-b和serpin-c分别在成虫期和蛹期表达量最高。serpin-a在中肠和围食膜中表达量最高,serpin-b在头、中肠和表皮中表达量最高,serpin-c在头部表达量最高,serpin-e在中肠和血淋巴中的表达量显著高于其他组织。棉铃虫取食低浓度Cry1Ac后,中肠的serpin-b和serpin-e的表达量显著增加。推测不同serpin基因在棉铃虫不同发育时期和组织中可能发挥不同的作用,其中serpin-b和serpin-e可能参与棉铃虫对Cry1Ac的解毒过程。     

棉铃虫BR-C Z2基因的克隆、原核表达及其表达谱

为明确转录因子广泛锌指复合物（broad complex,BR-C）在棉铃虫Helicoverpa armigera生长发育过程中的作用,基于转录组序列从棉铃虫幼虫中肠克隆获得BR-C Z2的cDNA序列并对其氨基酸序列和蛋白结构进行生物信息学分析,利用原核表达系统表达其融合蛋白;用实时荧光定量PCR（real-time fluorescence quantitative PCR,qRT-PCR）技术分析BR-C Z2基因在棉铃虫体内的表达规律,以及2-十三烷酮（2-tridecanone,2-TD）处理后其在棉铃虫6龄幼虫中肠内的变化规律。结果显示,棉铃虫BR-C Z2基因的开放阅读框为1 257 bp,编码418个氨基酸,预测编码蛋白的分子量和理论等电点分别为46.63 kD和6.94,主要定位于细胞核中。系统进化树结果显示棉铃虫BR-C Z2与家蚕Bombyx mori的BR-C Z2亲缘关系最近。成功表达His-HaBR-C Z2融合蛋白。BR-C Z2基因在棉铃虫蛹期和6龄幼虫中肠组织中相对表达量最高;不同浓度2-TD处理后,棉铃虫BR-C Z2基因相对表达量变化趋势不同,其中15 mg/g浓度处理12 h后棉铃虫BR-C Z2基因相对表达量达到峰值,是对照的2.5倍,而20 mg/g浓度处理20 h后棉铃虫BR-C Z2基因相对表达量降到最低,为对照的45.13%。表明BR-C Z2基因可能参与棉铃虫的生长发育,并响应2-TD的胁迫。     

Toxicity of some pyrethroids to the adult desert locust,Schistocerca gregaria forsk

The toxicity of a number of topically applied pyrethroids has been tested against adult male desert locusts, Schistocerca gregaria: the most potent proved to be 5-benzyl-3-furylmethyl (+)-trans-chrysanthemate (bioresmethrin) with a weighted mean LD₅₀ of 4.0 μg/g. The remaining compounds may be ranked in order of toxicity as follows: 5-benzyl-3-furylmethyl (±)-cis-trans-chrysanthemate (resmethrin) > 4-allyl-2,6-dimethyl-benzyl(+)-trans-chrysanthemate > 4-allylbenzyl (+)-trans-chrysanthemate > 2,4,6-trimethylbenzyl (+)-cis-trans-chrysanthemate > 2,3,4-trimethylbenzyl (+)-cis-trans-chrysanthemate > 2,4-dimethylbenzyl (±)-cis-trans-chrysanthemate; 2-methylbenzyl (±)-cis-trans-chrysanthemate. A small factor of synergism (4.2) was obtained with bioresmethrin following pre-treatment with sesamex, but with resmethrin the synergistic ratio (1.6) was of little practical significance.     

Acifluorfen metabolism in soybean: Diphenylether bond cleavage and the formation of homoglutathione, cysteine, and glucose conjugates

Metabolism of the substituted diphenylether herbicide, acifluorfen [sodium 5-(2-chloro-4-trifluoromethylphenoxy)-2-nitrobenzoate], was studied in excised leaf tissues of soybean [Glycine max (L.) Merr. ‘Evans’]. Studies with [chlorophenyl-¹⁴C]- and [nitrophenyl-¹⁴C]acifluorfen showed that the diphenylether bond was rapidly cleaved. From 85 to 95% of the absorbed [¹⁴C]acifluorfen was metabolized in less than 24 hr. Major polar metabolites were isolated and purified by solvent partitioning, adsorption, thin layer, and high-performance liquid chromatography. The major [chlorophenyl-¹⁴C]-labeled metabolite was identified as a malonyl-β- -glucoside (I) of 2-chloro-4-trifluoromethylphenol. Major [nitrophenyl-¹⁴C]-labeled metabolites were identified as a homoglutathione conjugate [S-(3-carboxy-4-nitrophenyl) γ-glutamyl-cysteinyl-β-alanine] (II), and a cysteine conjugate [S-(3-carboxy-4-nitrophenyl)cysteine] (III).