技术领域technical field
本发明属于烟草化学技术领域，具体涉及一种首次从烟草中提取得到的骈六元环联苯类化合物。同时本发明还涉及该化合物的制备方法和应用。The invention belongs to the technical field of tobacco chemistry, in particular to a parallel six-membered ring biphenyl compound extracted from tobacco for the first time. At the same time, the invention also relates to the preparation method and application of the compound.
背景技术Background technique
烟草是世界上化学成分最为复杂的植物，次生代谢产物非常丰富，经过几十年的研究，人们目前从烟草中鉴定出来的单体化学物质就超过3000多种，而且还有许多成分尚未鉴定出来。烟草除主要用于卷烟抽吸用途外，还可从中提取多种有利用价值的化学成分，从中发现有开发利用价值的先导性化合物。Tobacco is the plant with the most complex chemical composition in the world, and its secondary metabolites are very rich. After decades of research, more than 3,000 monomeric chemical substances have been identified from tobacco, and many components have not yet been identified. come out. In addition to being mainly used for smoking cigarettes, tobacco can also extract a variety of valuable chemical components from it, and discover leading compounds that are valuable for development and utilization.
联苯类化合物是一类天然植物中普遍存在的化合物，在烟草中也有存在该类化合物的文献报道，联苯类化合物具有广泛的药理作用，如抗肿瘤、抗人类免疫缺陷病毒(HIV)、抗氧化、抗菌、抗凝血等；同时已有研究证实，其药理作用与化学结构密切相关，可进一步研究和开发更多的联苯类化合物，从中寻找有效的先导化合和活性基团。本发明从烟草中分离得到了一种具有抗烟草花叶病毒活性的骈六元环联苯类化合物，该化合物至今尚未见到相关报道。Biphenyls are compounds that are ubiquitous in a class of natural plants, and there are also reports of such compounds in tobacco. Biphenyls have a wide range of pharmacological effects, such as anti-tumor, anti-human immunodeficiency virus (HIV), Antioxidant, antibacterial, anticoagulant, etc. At the same time, studies have confirmed that its pharmacological effects are closely related to its chemical structure, and more biphenyl compounds can be further studied and developed to find effective lead compounds and active groups. The present invention separates and obtains a parallel six-membered ring biphenyl compound with anti-tobacco mosaic virus activity from tobacco, and no related reports have been seen about the compound so far.
发明内容Contents of the invention
本发明的目的在于提供一种新的骈六元环联苯类化合物。The object of the present invention is to provide a new parallel six-membered ring biphenyl compound.
本发明的另一个目的是提供一种制备所述骈六元环联苯类化合物的方法；Another object of the present invention is to provide a method for preparing the parallel six-membered ring biphenyl compound;
本发明的目的还在于提供所述骈六元环联苯类化合物在制备抗烟草花叶病药物中的应用。The object of the present invention is also to provide the application of the parallel six-membered ring biphenyl compound in the preparation of anti-tobacco mosaic disease medicine.
本发明的目的通过下述技术方案予以实现。The purpose of the present invention is achieved through the following technical solutions.
除非另有说明，本发明中所采用的百分数均为质量百分数。Unless otherwise specified, the percentages used in the present invention are all mass percentages.
本发明所述的骈六元环联苯类化合物是从烟草(Nicotianatabacum)中分离得到，其分子式为C18H18O4，具有下述结构式：The parallel six-membered ring biphenyl compound of the present invention is isolated from tobacco (Nicotianatabacum), and its molecular formula is C18 H18 O4 , which has the following structural formula:
该化合物命名为烟草联苯C，英文名称为：TababiphenylC；为黄色胶状物。The compound is named Tobacco Biphenyl C, and its English name is: Tababiphenyl C; it is a yellow jelly.
一种制备所述骈六元环联苯类化合物的方法，以烟草为原料，经浸膏提取、硅胶柱层析、高压液相色谱、凝胶柱层析制备而成，具体包括以下步骤：A method for preparing the parallel six-membered ring biphenyl compound, using tobacco as a raw material, prepared through extraction from extract, silica gel column chromatography, high pressure liquid chromatography, and gel column chromatography, specifically comprising the following steps:
(1)浸膏提取：取烟草样品全株，将烟草粉碎或切成小段，用高浓度甲醇(w％：80％～100％)或高浓度乙醇(w％：80％～100％)或高浓度丙酮(w％：70％～100％)为提取溶剂，提取溶剂：烟草(重量比)＝2～4:1，浸泡24h～72h，提取3～5次，合并提取液、过滤浓缩成浸膏；(1) Extraction: Take the whole plant of tobacco sample, crush or cut the tobacco into small pieces, and use high-concentration methanol (w%: 80%-100%) or high-concentration ethanol (w%: 80%-100%) or High-concentration acetone (w%: 70% ~ 100%) is the extraction solvent, extraction solvent: tobacco (weight ratio) = 2 ~ 4:1, soak for 24h ~ 72h, extract 3 ~ 5 times, combine the extracts, filter and concentrate into extract;
(2)硅胶柱层析：浸膏用重量比1.5～3倍量的纯甲醇或纯乙醇或纯丙酮溶解后用重量比0.8～1.5倍的80～100目硅胶拌样，用重量比2～5倍量的160～300目硅胶干法装柱进行硅胶柱层析；以体积配比为(1:0、9:1、8:2、7:3、6:4、1:1和0:1)的氯仿-丙酮溶液进行梯度洗脱，合并相同的部分，收集各部分洗脱液并浓缩；(2) Silica gel column chromatography: the extract is dissolved with pure methanol or pure ethanol or pure acetone with a weight ratio of 1.5 to 3 times, and then mixed with 80 to 100 mesh silica gel with a weight ratio of 0.8 to 1.5 times. 5 times the amount of 160-300 mesh silica gel dry-packed column for silica gel column chromatography; the volume ratio is (1:0, 9:1, 8:2, 7:3, 6:4, 1:1 and 0 : 1) the chloroform-acetone solution carries out gradient elution, merges the same part, collects each part eluate and concentrates;
(3)高压液相色谱分离纯化：柱层析洗脱液的9:1部分进一步用高压液相色谱分离纯化即得所述的联苯类化合物。(3) Separation and purification by high-pressure liquid chromatography: the 9:1 part of the column chromatography eluent is further separated and purified by high-pressure liquid chromatography to obtain the biphenyl compounds.
高压液相色谱分离纯化是采用21.2mm×250mm，5μm的C18色谱柱，流速为20mL/min，流动相为43％的甲醇，紫外检测器检测波长为332nm，每次进样200μL，收集31.5min的色谱峰，多次累加后蒸干。The separation and purification of high-pressure liquid chromatography adopts a 21.2mm×250mm, 5μmC18 chromatographic column, the flow rate is 20mL/min, the mobile phase is 43% methanol, the detection wavelength of the ultraviolet detector is 332nm, and each injection is 200μL, collecting 31.5 The chromatographic peak of min was evaporated to dryness after being accumulated several times.
其中，凝胶柱层析步骤中，所得化合物再次用纯甲醇溶解，再以纯甲醇为流动相，用凝胶柱层析分离，以进一步分离纯化。Wherein, in the gel column chromatography step, the obtained compound is dissolved in pure methanol again, and then separated by gel column chromatography using pure methanol as a mobile phase for further separation and purification.
本发明的骈六元环联苯类化合物的结构通过以下方法测定。该化合物为黄色胶状物；紫外光谱(溶剂为甲醇),λmax(logε)213(4.26)、267(3.82)、332(3.38)nm；红外光谱(溴化钾压片)νmax3430,2923,2872,1675,1604,1542,1480,1435,1350,1138,962,871cm-1；高分辨质谱(HRESIMS)给出准分子离子峰m/z321.1106[M+Na]+(计算值321.1103)。结合1H和13CNMR谱给出一个分子式C18H18O4，不饱和度为10。从1H和13CNMR谱(数据归属见表-1)信号可以看出该化合物为联苯类化合物，其中可以明显发现有一个1,4-二取代的苯环、一个1,3,4,5-四取代苯环、一个甲氧基。详细对比该化合物与化合物的clusiparalicolineC(结构见图4)的核磁数据很接近，表明烟草联苯C是clusiparalicolineC的一个衍生物。The structure of the parallel six-membered ring biphenyl compound of the present invention is determined by the following method. The compound is yellow jelly; ultraviolet spectrum (solvent is methanol), λmax (logε) 213 (4.26), 267 (3.82), 332 (3.38) nm; infrared spectrum (potassium bromide tablet) νmax 3430, 2923,2872,1675,1604,1542,1480,1435,1350,1138,962,871cm-1 ; high-resolution mass spectrometry (HRESIMS) gives quasi-molecular ion peak m/z321.1106[M+Na]+ (calculated value 321.1103 ). Combining the1 H and13 CNMR spectra gave a molecular formula C18 H18 O4 with 10 degrees of unsaturation. From the signals of1 H and13 CNMR spectra (see Table-1 for data attribution), it can be seen that the compound is a biphenyl compound, in which it can be clearly found that there is a 1,4-disubstituted benzene ring, a 1,3,4, 5-tetrasubstituted benzene ring, a methoxyl group. A detailed comparison of the NMR data of this compound and the compound's clusiparalicolineC (see Figure 4 for the structure) is very close, indicating that tobacco biphenyl C is a derivative of clusiparalicolineC.
它们的主要不同在于烟草联苯C多了一个羰基、甲氧基和一个亚甲基，少了一对双键，这可能是由于C-7和C-8的变化导致。H-8到C-7、C-4、C-9、C-10和C-11的HMBC相关进一步证实了该推论。此外4'-OMe到C-4'的HMBC相关表明甲氧基与C-4'相连。至此本化合物的结构得以确定。Their main difference is that tobacco biphenyl C has one more carbonyl group, methoxy group and one methylene group, and one pair of double bonds is missing, which may be due to the changes of C-7 and C-8. The HMBC correlation of H-8 to C-7, C-4, C-9, C-10 and C-11 further confirmed this inference. Furthermore the HMBC correlation of 4'-OMe to C-4' indicates that the methoxy group is attached to C-4'. So far the structure of this compound has been determined.
表-1.化合物的1HNMR和13CNMR数据(CDCl3)Table-1.1 HNMR and13 CNMR data of the compound (CDCl3 )
将所述骈六元环联苯类化合物应用于制备抗烟草花叶病毒的药物。The parallel six-membered ring biphenyl compound is applied to the preparation of drugs against tobacco mosaic virus.
与现有技术相比，本发明具有以下优点：Compared with the prior art, the present invention has the following advantages:
本发明化合物系首次从烟草中分离提取得到的，通过核磁共振和质谱测定方法确定了为骈六元环联苯类化合物，并表征了其具体结构。经对抗烟草花叶病毒的实验，其相对抑制率达到48.4％，具有很好的抗烟草花叶病毒活性，超过阳性对照品南宁霉素的相对抑制率(29.6％)。以上结果揭示了本发明的化合物在制备抗烟草花叶病毒药物中有良好的应用前景。本发明化合物结构简单活性好，可作为抗烟草花叶病毒药物的先导性化合物。The compound of the present invention is isolated and extracted from tobacco for the first time, and is determined to be a parallel six-membered ring biphenyl compound through nuclear magnetic resonance and mass spectrometry methods, and its specific structure is characterized. Through the experiment against tobacco mosaic virus, its relative inhibition rate reaches 48.4%, and it has very good anti-tobacco mosaic virus activity, exceeding the relative inhibition rate (29.6%) of the positive control product Nanningmycin. The above results reveal that the compound of the present invention has good application prospects in the preparation of anti-tobacco mosaic virus drugs. The compound of the invention has a simple structure and good activity, and can be used as a lead compound of an anti-tobacco mosaic virus medicine.
附图说明Description of drawings
图1为本发明骈六元环联苯类化合物的核磁共振碳谱；Fig. 1 is the carbon nuclear magnetic resonance spectrum of parallel six-membered ring biphenyl compound of the present invention;
图2为本发明骈六元环联苯类化合物的核磁共振氢谱；Fig. 2 is the proton nuclear magnetic resonance spectrum of parallel six-membered ring biphenyl compound of the present invention;
图3为本发明骈六元环联苯类化合物的主要1H-1HCOSY和HMBC相关。Figure 3 shows the correlation between the main1 H-1 HCOSY and HMBC of the parallel six-membered ring biphenyl compounds of the present invention.
图4为对比化合物clusiparalicolineC的结构。Figure 4 is the structure of the comparative compound clusiparalicolineC.
具体实施方式detailed description
下面结合附图和实施例对本发明作进一步的详细说明，但不以任何方式对本发明加以限制，基于本发明教导所作的任何变换或改进，均落入本发明的保护范围。The present invention will be described in further detail below in conjunction with the accompanying drawings and examples, but the present invention is not limited in any way, and any changes or improvements based on the teaching of the present invention fall within the protection scope of the present invention.
实施例1Example 1
------化合物的制备，采用以下步骤：------The preparation of the compound adopts the following steps:
1、浸膏提取：取烟草样品全株，烟草可以粉碎或切成小段，用高浓度甲醇(w％：95％)或高浓度乙醇(w％：95％)或高浓度丙酮(w％：70％)为提取溶剂，提取溶剂：烟草(重量比)＝2～4:1，浸泡36h，提取4次，合并提取液、过滤浓缩成浸膏；1. Extraction of extract: get the whole plant of tobacco sample, tobacco can be pulverized or cut into small pieces, and use high concentration methanol (w%: 95%) or high concentration ethanol (w%: 95%) or high concentration acetone (w%: 95%) 70%) is the extraction solvent, extraction solvent: tobacco (weight ratio)=2～4:1, soak for 36h, extract 4 times, combine extract, filter and concentrate to become medicinal extract;
2、硅胶柱层析：浸膏用重量比2.5倍量的纯甲醇或纯乙醇或纯丙酮溶解后用重量比1.5倍的80～100目硅胶拌样，用重量比2.5倍量的160～300目硅胶干法装柱进行硅胶柱层析；以体积配比为(1:0、9:1、8:2、7:3、6:4、1:1、0:1)的氯仿-丙酮溶液进行梯度洗脱，合并相同的部分，收集各部分洗脱液并浓缩；2. Silica gel column chromatography: dissolve the extract with 2.5 times the weight of pure methanol or pure ethanol or pure acetone, mix the sample with 1.5 times the weight of 80-100 mesh silica gel, and use 2.5 times the weight of 160-300 Silica gel dry packing for silica gel column chromatography; chloroform-acetone with a volume ratio of (1:0, 9:1, 8:2, 7:3, 6:4, 1:1, 0:1) The solution is subjected to gradient elution, the same fractions are combined, and the eluents of each fraction are collected and concentrated;
3、高压液相色谱分离：柱层析洗脱液的9:1部分进一步用高压液相色谱分离纯化即得所述的联苯类化合物，高压液相色谱分离纯化是采用21.2mm×250mm，5μm的C18色谱柱，流速为20mL/min，流动相为43％的甲醇，紫外检测器检测波长为332nm，每次进样200μL，收集31.5min的色谱峰，多次累加后蒸干。3. High-pressure liquid chromatography separation: the 9:1 part of the column chromatography eluent is further separated and purified by high-pressure liquid chromatography to obtain the described biphenyl compounds. The high-pressure liquid chromatography separation and purification is 21.2mm×250mm, 5 μm C18 chromatographic column, the flow rate is 20mL/min, the mobile phase is 43% methanol, the detection wavelength of the ultraviolet detector is 332nm, each injection is 200μL, the chromatographic peaks are collected for 31.5min, and evaporated to dryness after multiple accumulations.
4、凝胶柱层析：步骤3中高压液相色谱法分离纯化后的物质经所述高效液相色谱分离纯化后，所得化合物再次用纯甲醇溶解，再以纯甲醇为流动相，用凝胶柱层析分离，以进一步分离纯化。4. Gel column chromatography: After the material separated and purified by high-pressure liquid chromatography in step 3 is separated and purified by high-performance liquid chromatography, the obtained compound is dissolved in pure methanol again, and then pure methanol is used as mobile phase, and gel Column chromatography separation for further separation and purification.
本发明所用原料不受地区和品种限制，均可以实现本发明，下面以来源于云南中烟工业有限责任公司不同产地的烟草原料对本发明做进一步说明：The raw materials used in the present invention are not limited by regions and varieties, and the present invention can be realized. The present invention will be further described below with tobacco raw materials derived from different places of production of China Tobacco Yunnan Industry Co., Ltd.:
实施例2Example 2
烟草样品来源于云南昆明，品种为红花大金元。将烟草取样2.0kg粉碎以95％的甲醇提取5次，每次提取24h，提取液合并，过滤，减压浓缩成浸膏，得浸膏100g。浸膏用重量比2.0倍量的纯甲醇溶解后用120g的100目粗硅胶拌样，0.6kg的160目硅胶装柱进行硅胶柱层析，用体积配比为1:0、9:1、8:2、7:3、6:4、1:1、0:1的氯仿-丙酮梯度洗脱，TLC监测合并相同的部分，得到8个部分，其中体积配比为9:1的氯仿-丙酮洗脱部分用安捷仑1100半制备高效液相色谱分离，以43％的甲醇为流动相，ZorbaxSB-C18(21.2×250mm,5μm)制备柱为固定相，流速为20ml/min，紫外检测器检测波长为332nm，每次进样200μL，收集31.5min的色谱峰，多次累加后蒸干；所得产物再次用纯甲醇溶解，再以纯甲醇为流动相，用SephadexLH-20凝胶柱层析分离，即得该新化合物。The tobacco samples were from Kunming, Yunnan, and the variety was Honghua Dajinyuan. Sampling 2.0 kg of tobacco was pulverized and extracted 5 times with 95% methanol for 24 hours each time. The extracts were combined, filtered, and concentrated under reduced pressure to form an extract to obtain 100 g of extract. After dissolving the extract with 2.0 times the amount of pure methanol by weight, mix the sample with 120g of 100-mesh silica gel, pack 0.6kg of 160-mesh silica gel into a column for silica gel column chromatography, and use a volume ratio of 1:0, 9:1, 8:2, 7:3, 6:4, 1:1, 0:1 chloroform-acetone gradient elution, TLC monitoring and merging the same parts to obtain 8 parts, in which the volume ratio is 9:1 chloroform-acetone The acetone elution part was separated by Agilent 1100 semi-preparative high-performance liquid chromatography, with 43% methanol as mobile phase, ZorbaxSB-C18 (21.2×250mm, 5μm) preparative column as stationary phase, flow rate was 20ml/min, UV detection The detection wavelength of the detector is 332nm, each injection is 200μL, and the chromatographic peak is collected for 31.5min, and evaporated to dryness after repeated accumulation; Separation, that is, the new compound.
实施例3Example 3
烟草样品来源于云南大理，品种为K326，将烟草取样4.0kg切碎，以95％的乙醇提取4次，每次提取48h，提取液合并，过滤，减压浓缩成浸膏，得浸膏192g。浸膏用重量比2.0倍量的纯甲醇溶解后用200g的80目粗硅胶拌样，1.2kg的200目硅胶装柱进行硅胶柱层析，用体积配比为1:0、9:1、8:2、7:3、6:4、1:1、0:1的氯仿-丙酮梯度洗脱，TLC监测合并相同的部分，得到8个部分，其中体积配比为9:1的氯仿-丙酮洗脱部分用安捷仑1100半制备高效液相色谱分离，以43％的甲醇为流动相，ZorbaxSB-C18(21.2×250mm,5μm)制备柱为固定相，流速为20ml/min，紫外检测器检测波长为332nm，每次进样200μL，收集31.5min的色谱峰，多次累加后蒸干；所得产物再次用纯甲醇溶解，再以纯甲醇为流动相，用SephadexLH-20凝胶柱层析分离，即得该新化合物。The tobacco sample comes from Dali, Yunnan, and the variety is K326. The tobacco sample is 4.0kg and chopped, extracted 4 times with 95% ethanol, each extraction is 48h, the extracts are combined, filtered, and concentrated under reduced pressure to obtain an extract 192g . The extract is dissolved in pure methanol with a weight ratio of 2.0 times, and then mixed with 200 g of 80 mesh thick silica gel, and 1.2 kg of 200 mesh silica gel is packed into a column for silica gel column chromatography, and the volume ratio is 1:0, 9:1, 8:2, 7:3, 6:4, 1:1, 0:1 chloroform-acetone gradient elution, TLC monitoring and merging the same parts to obtain 8 parts, in which the volume ratio is 9:1 chloroform-acetone The acetone eluted part was separated by Agilent 1100 semi-preparative high-performance liquid chromatography, with 43% methanol as mobile phase, ZorbaxSB-C18 (21.2×250mm, 5μm) preparative column as stationary phase, flow rate was 20ml/min, UV detection The detection wavelength of the detector is 332nm, each injection is 200μL, and the chromatographic peak is collected for 31.5min, and evaporated to dryness after repeated accumulation; Separation, that is, the new compound.
实施例4Example 4
烟草样品来源于云南楚雄，品种为云烟200，将烟草取样6kg粉碎，以75％的丙酮用超声提取3次，每次提取72h，提取液合并，过滤，减压浓缩成浸膏，得浸膏320g。浸膏用重量比1.6倍量的纯甲醇溶解后用360g的90目粗硅胶拌样，2.4kg的180目硅胶装柱进行硅胶柱层析，用体积配比为1:0、9:1、8:2、7:3、6:4、1:1、0:1的氯仿-丙酮梯度洗脱，TLC监测合并相同的部分，得到8个部分，其中体积配比为9:1的氯仿-丙酮洗脱部分用安捷仑1100半制备高效液相色谱分离，以43％的甲醇为流动相，ZorbaxSB-C18(21.2×250mm,5μm)制备柱为固定相，流速为20ml/min，紫外检测器检测波长为332nm，每次进样200μL，收集31.5min的色谱峰，多次累加后蒸干；所得产物再次用纯甲醇溶解，再以纯甲醇为流动相，用SephadexLH-20凝胶柱层析分离，即得该新化合物。The tobacco sample comes from Chuxiong, Yunnan, and the variety is Yunyan 200. The tobacco sample is 6kg and pulverized, and extracted 3 times with 75% acetone by ultrasound, each time for 72 hours, the extracts are combined, filtered, and concentrated under reduced pressure to obtain an extract. 320g. The extract is dissolved in pure methanol with a weight ratio of 1.6 times, and then mixed with 360 g of 90 mesh thick silica gel, and 2.4 kg of 180 mesh silica gel is packed for silica gel column chromatography, and the volume ratio is 1:0, 9:1, 8:2, 7:3, 6:4, 1:1, 0:1 chloroform-acetone gradient elution, TLC monitoring and merging the same parts to obtain 8 parts, in which the volume ratio is 9:1 chloroform-acetone The acetone eluted part was separated by Agilent 1100 semi-preparative high-performance liquid chromatography, with 43% methanol as mobile phase, ZorbaxSB-C18 (21.2×250mm, 5μm) preparative column as stationary phase, flow rate was 20ml/min, UV detection The detection wavelength of the detector is 332nm, each injection is 200μL, and the chromatographic peak is collected for 31.5min, and evaporated to dryness after repeated accumulation; Separation, that is, the new compound.
实施例5Example 5
-------化合物的鉴定-------identification of compounds
取实施例2制备的化合物，为黄色胶状物。紫外光谱(溶剂为甲醇),λmax(logε)213(4.26)、267(3.82)、332(3.38)nm；红外光谱(溴化钾压片)νmax3430,2923,2872,1675,1604,1542,1480,1435,1350,1138,962,871cm-1；高分辨质谱(HRESIMS)给出准分子离子峰m/z321.1106[M+Na]+(计算值321.1103)。结合1H和13CNMR谱给出一个分子式C18H18O4，不饱和度为10。从1H和13CNMR谱(数据归属见表-1)信号可以看出该化合物为联苯类化合物，其中可以明显发现有一个1,4-二取代的苯环、一个1,3,4,5-四取代苯环、一个甲氧基。详细对比该化合物与化合物的clusiparalicolineC(结构见图4)的核磁数据很接近，表明烟草联苯C是clusiparalicolineC的一个衍生物。The compound prepared in Example 2 was a yellow jelly. UV spectrum (solvent is methanol), λmax (logε) 213 (4.26), 267 (3.82), 332 (3.38) nm; infrared spectrum (potassium bromide tablet) νmax 3430, 2923, 2872, 1675, 1604, 1542,1480,1435,1350,1138,962,871cm-1 ; high resolution mass spectrometry (HRESIMS) gave quasi-molecular ion peak m/z 321.1106[M+Na]+ (calculated value 321.1103). Combining the1 H and13 CNMR spectra gave a molecular formula C18 H18 O4 with 10 degrees of unsaturation. From the signals of1 H and13 CNMR spectra (see Table-1 for data attribution), it can be seen that the compound is a biphenyl compound, in which it can be clearly found that there is a 1,4-disubstituted benzene ring, a 1,3,4, 5-tetrasubstituted benzene ring, a methoxyl group. A detailed comparison of the NMR data of this compound and the compound's clusiparalicolineC (see Figure 4 for the structure) is very close, indicating that tobacco biphenyl C is a derivative of clusiparalicolineC.
它们的主要不同在于烟草联苯C多了一个羰基、甲氧基和一个亚甲基，少了一对双键，这可能是由于C-7和C-8的变化导致。H-8到C-7、C-4、C-9、C-10和C-11的HMBC相关进一步证实了该推论。此外4'-OMe到C-4'的HMBC相关表明甲氧基与C-4'相恋。至此本化合物的结构得以确定。Their main difference is that tobacco biphenyl C has one more carbonyl group, methoxy group and one methylene group, and one pair of double bonds is missing, which may be due to the changes of C-7 and C-8. The HMBC correlation of H-8 to C-7, C-4, C-9, C-10 and C-11 further confirmed this inference. Furthermore the HMBC correlation of 4'-OMe to C-4' suggests that the methoxy group is in love with C-4'. So far the structure of this compound has been determined.
实施例6Example 6
取实施例3制备的化合物，为黄色胶状物。测定方法与实施例5相同，确认实施例3制备的化合物为所述的联苯类化合物——烟草联苯C。The compound prepared in Example 3 was a yellow jelly. The determination method was the same as in Example 5, and it was confirmed that the compound prepared in Example 3 was the biphenyl compound—tobacco biphenyl C.
实施例7Example 7
取实施例4制备的化合物，为黄色胶状物。测定方法与实施例5相同，确认实施例4制备的化合物为所述的联苯类化合物——烟草联苯C。Get the compound prepared in Example 4, which is a yellow jelly. The determination method was the same as in Example 5, and it was confirmed that the compound prepared in Example 4 was the biphenyl compound—tobacco biphenyl C.
实施例8Example 8
------抗烟草花叶病毒的活性试验------Anti-tobacco mosaic virus activity test
取实施例2～4制备的任一联苯类化合物进行抗烟草花叶病毒活性试验，试验情况如下：Get any biphenyl compounds prepared in Examples 2 to 4 to carry out the anti-tobacco mosaic virus activity test, and the test conditions are as follows:
采用半叶法，在药剂的质量浓度均为50mg/L时对本发明化合物进行抗烟草花叶病毒活性测定。在5～6龄烤烟的植株上，选取适用于测试的叶片(叶行正常，无病无虫)，先将叶片均匀撒上细金刚砂，用毛笔将备用的烟草花叶病毒源(3.0×10-3)均匀抹在撒有金刚砂的叶片上，待所有中选的叶片接毒结束后，立即放在盛有药液的培养皿中处理20min，取出，擦去叶片上水珠和药液，将两个半叶复原排放在铺有卫生纸保湿的玻璃缸中，并盖上玻璃盖，控温(23±2)℃，放在温室自然光照射，2～3d即可见枯斑.每个处理都设另一半叶为对照，另外设有1组为商品宁南霉素的处理作为对比，按下公式计算相对抑制率。The anti-tobacco mosaic virus activity of the compound of the present invention was determined when the mass concentration of the medicament was 50 mg/L by half-leaf method. On the plants of flue-cured tobacco in the 5th to 6th age, select the leaves suitable for the test (the leaf row is normal, no disease and no insects), and the leaves are evenly sprinkled with fine emery, and the spare tobacco mosaic virus source (3.0 × 10-3 ) Spread evenly on the leaves sprinkled with carborundum. After all the selected leaves are inoculated with the poison, immediately place them in a petri dish containing the medicine solution for 20 minutes, take them out, wipe off the water drops and medicine solution on the leaves, and put The two half-leaves were restored and placed in a glass jar covered with toilet paper and covered with a glass lid. The temperature was controlled at (23±2)°C, and they were placed in the greenhouse under natural light. Blight spots could be seen after 2-3 days. The other half of the leaf was the control, and another group was treated with commercial Ningnanmycin as a comparison, and the relative inhibition rate was calculated according to the formula.
XI％＝(CK-T)/CK×100％XI%=(CK-T)/CK×100%
X：相对抑制率(％)，CK：浸泡于清水中半片接毒叶的枯斑数(个)，T浸泡于药液中半片接毒叶的枯斑数(个)。X: relative inhibition rate (%), CK: the number of dead spots (pieces) of half a poisoned leaf soaked in clear water, T the number of dead spots (pieces) of half a poisoned leaf soaked in medicinal liquid.
结果明本化合物的相对抑制率为48.4％，超过对照宁南霉素的相对抑制率29.6％，说明化合物具有很好的抗烟草花叶病毒活性。The results showed that the relative inhibition rate of the compound was 48.4%, which was higher than that of the control Ningnanmycin, which was 29.6%, indicating that the compound had good activity against tobacco mosaic virus.
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| CN105175383B (en)* | 2015-08-12 | 2017-04-05 | 云南民族大学 | A kind of biphenyl compound and its preparation method and application |
| CN109438464B (en)* | 2018-11-19 | 2020-06-16 | 云南中烟工业有限责任公司 | A kind of chromone derivative extracted from Cassia brevis and its preparation method and application |
| CN117186049B (en)* | 2023-09-07 | 2025-07-22 | 贵州大学 | 2, 2-Dimethyl-2H chromene derivative containing sulfonamide unit, and preparation method and application thereof |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2145082A (en)* | 1983-08-15 | 1985-03-20 | Erba Farmitalia | Process for the preparation of 2,3-dihydro-[4H)-1 -benzopyran-4-one and 2,3,5,6,7,8- hexahydro-(4H)-1-benzopyran-4,5-dione derivatives |
| CN102090412A (en)* | 2009-12-11 | 2011-06-15 | 南开大学 | Ferulic acid and ferulic acid derivative anti-phytoviral agents |
| CN102304114A (en)* | 2011-08-08 | 2012-01-04 | 云南省烟草农业科学研究院 | Flavanone compound and application thereof |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2145082A (en)* | 1983-08-15 | 1985-03-20 | Erba Farmitalia | Process for the preparation of 2,3-dihydro-[4H)-1 -benzopyran-4-one and 2,3,5,6,7,8- hexahydro-(4H)-1-benzopyran-4,5-dione derivatives |
| CN102090412A (en)* | 2009-12-11 | 2011-06-15 | 南开大学 | Ferulic acid and ferulic acid derivative anti-phytoviral agents |
| CN102304114A (en)* | 2011-08-08 | 2012-01-04 | 云南省烟草农业科学研究院 | Flavanone compound and application thereof |
| Title |
|---|
| New Biphenyl Compounds with DNA Strand-Scission Activity from Clusia paralicola;Eun-Kyoung Seo et al.;《J. Nat. Prod.》;19991002;第62卷(第11期);第1484-1487页* |
| 宁南霉素对烟草花叶病毒的生物活性;邓光兵等;《应用与环境生物学报》;20041225;第10卷(第6期);第695-698页* |
| Publication number | Publication date |
|---|---|
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| Publication | Publication Date | Title |
|---|---|---|
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