A new phenylethanoid triglycoside in Veronica beccabunga L

Besides the expected iridoid glucosides aucubin and catalpol as well as three known esters of the latter, Veronica beccabunga (brooklime) was shown to contain five carboxylated iridoid glucosides, namely gardoside, mussaenosidic acid, 8-epiloganic acid, arborescosidic acid and alpinoside. In addition to these compounds, the plant contained salidroside and a previously unknown caffeoyl phenylethanoid glycoside (CPG) which we have named chionoside J. The structure was elucidated mainly by 1D and 2D NMR spectroscopy to be 2″-(β-glucopyranosyl)-plantamajoside. The distribution of plantamajoside and its derivatives as well as that of carbocyclic iridoids with an 8,9-double bond is briefly discussed, and it is noted that such compounds are mainly confined to the tribe Veroniceae of the Plantaginaceae.     

Chemical markers in Veronica sect. Hebe. III

In a continued chemosystematic investigation of the water-soluble compounds in Veronica sect. Hebe, we have investigated two more of the species formerly classified as Parahebe. Both species contained mannitol in considerable amounts and in addition some glucosides of iridoid acids. Veronica cheesemanii was characterised by aucubin and its esters: 2′-O-benzoylaucubin and an aucubin diester named cheesemanioside. The main iridoid compounds in Veronica hookeriana were catalpol and its ester verminoside, but this species also contained the sugar ester methyl 1-O-benzoyl-3-α-glucuronosylglycerol and a caffeoyl phenylethanoid glycoside (CPG) named parahebeoside, a 2′-O-β-xylopyranosyl derivative of the known plantamajoside. The results show that the studied species of the former genus Parahebe are very different with regard to their chemical content. This is in agreement with the DNA sequence data and implies the genus was polyphyletic as previously circumscribed.     

Chemotaxonomy of Veroniceae and its allies in the Plantaginaceae

In a chemosystematic investigation of tribe Veroniceae (Plantaginaceae), representatives of Camptoloma, Sibthorpia, Veronica subg. Pentasepalae and subg. Hebe, Veronicastrum, Wulfenia, and the related Ellisiophyllum and Globularia were examined for non-flavonoid glycosides. From the 14 species studied, 28 different iridoid glucosides and 10 caffeoyl phenylethanoid glucosides (CPGs), as well as salidroside and arbutin were isolated and characterized by NMR; of these, five compounds were previously unknown. It was found that the representatives of Veroniceae, as well as Globularia, were characterized by mannitol, aucubin, catalpol and catalpol esters. Each of the three studied species of Veronica subg. Hebe contained at least one of the 6-O-catalpol esters typical for Veronica s. str. (verminoside), supporting the inclusion of Hebe in Veronica. However, their main constituents were esters of 6-O-rhamnopyranosylcatalpol; a CPG, hebeoside (2'-beta-xylopyranosyl-verbascoside) was isolated from V. (Hebe) salicifolia. The two species of Veronicastrum also contained 6-O-rhamnopyranosylcatalpol esters, including the previously unknown 2',3'- and 3',4'-dicinnamoyl derivatives and, in contrast to the earlier reports, they lacked 6-O-catalpol esters. The main iridoid constituents in the three investigated species of Wulfenia were 10-O-aucubin and 10-O-catalpol esters (isoscrophularioside or globularin) while baldaccioside (10-O-cinnamoyl asystasioside E) was isolated from W. baldaccii. Globularia vulgaris contained 10-O-catalpol esters (e.g., globularin) and, in addition, asperuloside together with its benzoyl analogue named besperuloside. The representatives of Sibthorpia and Ellisiophyllum were almost completely devoid of iridoids; this, however, together with the CPGs present implied a close relationship between the two genera. Camptoloma lyperiiflorum lacked hexitols but contained esters of 6-O-rhamnopyranosylcatalpol different from those found in Veroniceae but known from Buddleja, Scrophularia and Verbascum (Scrophulariaceae s. str.).     

Iridoids and phenylethanoids in Lagotis integrifolia and Wulfeniopsis amherstiana (Plantaginaceae)

In a chemosystematic investigation of Wulfeniopsis amherstiana we have isolated four common iridoid glucosides as well as the esters 3″- and 4″-Cinnamoyl 6-O-rhamnopyranosylcatalpol. Furthermore the plant contained mannitol, arbutin, and four caffeoyl phenylethanoid glucosides, namely plantamajoside, aragoside and two new acetyl derivatives of the latter, named amherstianoside A and B. From Lagotis integrifolia we have obtained mannitol and three common iridoid glucosides together with the 8,9-unsaturated iridoids arborescosidic acid and anagalloside. The results show that the genus Wulfeniopsis is chemically different from Wulfenia and that it is closer related to Veronicastrum.     

Chemical markers in Veronica sect. Hebe. II

In a continued chemosystematic investigation of the water-soluble compounds in Veronica sect. Hebe, four additional species were investigated. In comparison to other, Northern Hemisphere (NH) species of Veronica, those belonging to the New Zealand species in sect. Hebe are apparently more variable in chemical content. In addition to the compounds characteristic for NH Veronica, namely mannitol, aucubin, catalpol and 6-O-esters of catalpol as well as some caffeoyl phenylethanoid glucosides (CGPs), Veronica topiaria (syn. Hebe topiaria) also gave an unusual 6-O-ester of aucubin named topiarioside. The former Hebe species Veronica cupressoides and Veronica stenophylla each provided one of the two previously undescribed disaccharide esters named hebitol I and II, respectively, and the former plant also provided a CPG named cuproside, a 6-O-β-glucopyranosyl derivative of the known hebeoside. The last species, namely Veronica hulkeana (syn. Heliohebe hulkeana) only contained compounds common to other species of Veronica. The taxonomic results are discussed and it is concluded that carbohydrate esters are common in sect. Hebe. The data so far obtained indicate that the occurrences of esters of 6-O-rhamnopyranosylcatalpol are confined to the most derived species in the section.     

Unusual iridoid glycosides in Veronica sects. Hebe and Labiatoides

In a chemosystematic investigation of three Southern hemisphere species of Veronica, namely the Australian Veronica derwentiana Andrews and Veronica perfoliata R.Br. (formerly Derwentia species), and the New Zealand Veronica catarractae G. Forster (formerly a species of Parahebe), the water-soluble constituents were isolated and identified by spectroscopic methods. Apart from other iridoid glucosides common to the genus, three unusual substituted benzoyl esters of aucubin (derwentiosides A–C) were obtained from V. derwentiana and a chlorinated iridoid glycoside (catarractoside) from V. catarractae in addition to other iridoids common to the genus. The chemical profile of V. perfoliata is similar to that of Northern hemisphere species of Veronica because of the presence of characteristic 6-O-catalpol esters. The profile of V. derwentiana is unique, since 6-O-esters of aucubin rather than of catalpol dominate, however, the acyl groups are the same as those present in catalpol esters found in some other Veronica sections. V. catarractae also contains one of the catalpol esters characteristic of Veronica, but in addition three 6-O-rhamnopyranosyl substituted iridoid glycosides, one of which is 6-O-rhamnopyranosylcatalpol. Esters of the latter compound are previously only known from the more derived species in recent phylogenetic trees of sect. Hebe to which V. catarractae now also belongs, but as a more basal member.     

Chemotaxonomic markers in Digitalideae (Plantaginaceae)

In a chemosystematic investigation of Digitalideae (Plantaginaceae), the water-soluble part of extracts of two species of Digitalis, two species of Isoplexis, as well as Erinus alpinus and Lafuentea rotundifolia were studied with regard to their content of main carbohydrates, iridoids and caffeoyl phenylethanoid glycosides (CPGs). Digitalis and Isoplexis contained sorbitol, cornoside and a number of other phenylethanoid glycosides including the new tyrosol beta-D-mannopyranoside, sceptroside but were found to lack iridoid glucosides. Erinus contained mainly glucose, the new 8,9-double bond iridoid, erinoside, and a number of known iridoid glucosides including two esters of 6-rhamnopyranosylcatalpol, as well as the CPG poliumoside. Finally, Lafuentea was characterized by the presence of glucose, aucubin and cryptamygin B but apparently lacked CPGs. The chemosystematic significance of the isolated compounds is discussed.     

Iridoids from Scutellaria albida ssp. albida

Three iridoid glycosides, 6'-O-E-p-coumaroylgardoside (1), 6'-O-p-E-coumaroyl-8-epi-loganic acid (2) and scutelloside (3) were isolated from the aerial parts of Scutellaria albida subsp. albida, in addition to an anomeric mixture in equilibrium of one iridoid aglycone (4, 4a), nine iridoid glycosides (5-13), four known phenylethanoid glycosides (14-17), and six known phenolic derivatives (18-23).     

Characterization of enzyme-catalysed endogenous β-hydroxylation of phenylethanoid glycosides in Euphrasia rostkoviana Hayne at the molecular level

Phenylethanoid glycosides, the main constituents of the aerial part of eyebright (Euphrasia rostkoviana Hayne) were treated by the endogenous hydroxylase enzyme and the concomitant biotransformation was characterized by applying high-performance liquid chromatography with UV and MS detections and NMR spectroscopy. In the extracts of the untreated (intact) samples, acteoside and eukovoside were determined as main compounds. The enzymatic treatment resulted in the quantitative transformation of these phenylethanoid glycosides into their corresponding hydroxyl derivatives identified as two epimers of β-hydroxyacteoside and β-hydroxyeukovoside. As to the importance of this hydroxylation β-hydroxyeukovoside was identified as a new compound and β-hydroxyacteoside was described for the first time in eyebright. We proved for first time that a β-hydroxylase enzyme is active in eyebright tissues which can transform phenylethanoid glycosides into their β-hydroxyl derivatives. Our new enzymatic method combined with a preparative HPLC facilitates the isolation of β-hydroxyl phenylethanoid glycosides from the aerial part of eyebright.     

Veronica: Iridoids and cornoside as chemosystematic markers

The iridoid glucoside, ajugol, and the phenylethanoid glucoside, cornoside, have been isolated from species of Veronica (Plantaginaceae) for the first time. The presence of these compounds has been screened in 18 plant accessions belonging to 15 species of Veronica (Plantaginaceae), by isolation or NMR spectroscopy of crude extracts. In addition, the distribution of iridoids in the genus has been reviewed, using mainly the published data of isolated compounds. Using the recent expansion and reclassification of the genus based on DNA-sequence results as the model, we find that the genus is rather homogeneous with regard to the distribution of iridoid glucosides, aucubin and/or catalpol as well as 6-O-esters of catalpol being universally present in 10 of the 12 subgenera for which data exist. Only the two subgenera Pocilla and Chamaedrys deviate from this pattern. Pocilla is heterogeneous; in this subgenus, species in subsect. Agrestes contain the standard iridoid garniture, while species in subsect. Biloba do not contain the 6-O-esters of catalpol, but ajugol instead. Veronica intercedens (subsect. Subracemosae) differs from the remainder of the subgenus in only containing 5-hydroxylated iridoids (melittoside and globularifolin) and is so far the only species within the genus in which such compounds have been detected. These chemical differences are clearly reflected in the DNA-based phylogram of the subgenus. Subg. Chamaedrys appears homogeneous in lacking iridoids or only containing these in small amounts, but instead half of the investigated species contained the phenylethanoid glucoside cornoside. The distribution of this compound in angiosperms is reviewed; cornoside often substitutes iridoid glucosides in plants where these are expected to be present. The chemical results of Veronica fit in very well with the phylogenetic implications of the DNA-sequence results.     

Phenylethanoid glycosides from Lippia javanica

Lippia javanica (N.L.Burm.) Spreng. is an aromatic, multipurpose medicinal plant from which a number of volatile compounds have been identified, together with toxic triterpenoids and iridoid glycosides. Two additional phenylethanoid glycosides, verbascoside and isoverbascoside, were isolated from L. javanica and characterized. High performance liquid chromatography analyses of polar extracts of three other Lippia species (L. scaberrima, L. rehmannii and L. wilmsii), indigenous to South Africa, revealed the presence of both isomers. When compared to the other indigenous Lippia species, the leaves of L. javanica were found to contain the highest concentrations of both isomers. In addition, the intraspecies variation of the verbascoside/isoverbascoside content of L. javanica, harvested from the same and different localities, was investigated. The concentrations of the two phenylethanoids remained fairly consistent within and between different populations, even when geographically separated. While these compounds are produced by many genera, they may now be added to the list of iridoid glucosides employed as chemotaxonomic markers for Lippia species.     

Tyrosinase Inhibitors from the Aerial Parts of Wulfenia carinthiaca Jacq.

Activity guided isolation of a MeOH extract of the aerial plant parts of Wulfenia carinthiaca Jacq . (Plantaginaceae), using a mushroom tyrosinase assay, resulted in the isolation of five phenylethanoid glucosides and four iridoid glycosides. Two of them, 2′‐O‐acetylisoplantamajoside and 2′,6″‐O‐diacetylisoplantamajoside, represent new natural products. Evaluation of the inhibitory activity of all isolated compounds revealed that the observed activity is not related to the isolated phenylethanoid glycosides but mainly due to the presence of the iridoid glycoside globularin (IC₅₀ 41.94 μm ; CI_95% ± 16.61/11.89 μm ). Interestingly, structurally close related compounds (globularicisin, baldaccioside, and isoscrophularioside) showed no or only a weak tyrosinase inhibitory activity.     

Melanogenesis‐Inhibitory Saccharide Fatty Acid Esters and Other Constituents of the Fruits of Morinda citrifolia (Noni)

Five new saccharide fatty acid esters, named nonioside P ( 3 ), nonioside Q ( 4 ), nonioside R ( 8 ), nonioside S ( 10 ), and nonioside T ( 14 ), and one new succinic acid ester, butyl 2‐hydroxysuccinate (=4‐butoxy‐3‐hydroxy‐4‐oxobutanoic acid) ( 31 ), were isolated, along with 26 known compounds, including eight saccharide fatty acid esters, 1, 2, 5, 6, 7, 9, 12 , and 13 , three hemiterpene glycosides, 15, 17 , and 18 , six iridoid glycosides, 21 – 25 , and 27 , and nine other compounds, 20, 28, 29 , and 32 – 37 , from a MeOH extract of the fruit of Morinda citrifolia (noni). Upon evaluation of these and five other glycosidic compounds, 11, 16, 19, 26 , and 30 , from M. citrifolia fruit extract for their inhibitory activities against melanogenesis in B16 melanoma cells induced with α‐melanocyte‐stimulating hormone (α‐MSH), most of the saccharide fatty acid esters, hemiterpene glycosides, and iridoid glycosides showed inhibitory effects with no or almost no toxicity to the cells. These compounds were further evaluated with respect to their cytotoxic activities against two human cancer cell lines (HL‐60 and AZ521) and their inhibitory effects on Epstein? Barr virus early antigen (EBV‐EA) activation induced with 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) in Raji cells.     

Chemotaxonomy of Plantago. Iridoid glucosides and caffeoyl phenylethanoid glycosides

Data for 34 species of Plantago (Plantaginaceae), including subgen. Littorella (= Littorella uniflora), have been collected with regard to their content of iridoid glucosides and caffeoyl phenylethanoid glycosides (CPGs). In the present work, 21 species were investigated for the first time and many known compounds were found together with three new iridoid glucosides. Of these, arborescoside and arborescosidic acid, both of the uncommon type with an 8,9-double bond, were present in several species, while 6-deoxymelittoside was found only in P. subulata. The known compounds deoxyloganic acid, caryoptoside and rehmannioside D were isolated from the genus for the first time. The earlier reported occurrence of sorbitol in the family was confirmed, and this compound was shown by NMR spectroscopy to be the main sugar in the three species investigated for this. The combined data show that CPGs are present in all species investigated. With regard to the iridoids, the distribution patterns showed a good correlation with the classification of Rahn. Thus, aucubin is typical for the whole genus, while bartsioside and catalpol as well as 5-substituted iridoids are each characteristic for a subgenus in the family. Finally, the close relationship between Plantago and Veronica suggested by chloroplast DNA sequence analysis. could be corroborated by the common occurrence of the rare 8,9-unsaturated iridoids in these two genera.     

Potential benefits of iridoid glycoside sequestration in Longitarsus melanocephalus (Coleoptera,Chrysomelidae)

Whenever potentially noxious plant compounds are taken up and recycled by herbivorous insects, a protective function of these sequestered compounds is assumed. The flea beetle Longitarsus melanocephalus sequesters iridoid glycosides from its host plant up to a concentration of 2% DW, yet so far it remained unknown whether the insects gain protection from natural enemies by sequestering plant compounds at these comparatively low concentrations. Here we tested whether iridoid glycosides might deter or inhibit various soil and litter-dwelling potential enemies and pathogens. In choice experiments presenting L. melanocephalus pupae together with Tribolium castaneum pupae, the predator Lithobius forficatus was deterred by the iridoid glycoside containing pupae, while Forficula auricularia as well as the nematode Heterorhabditis bacteriophora were not deterred. L. forficatus also avoided artificial baits doted with 2% iridoid glycosides while F. auricularia showed no aversion to iridoid glycosides at these concentrations and H. bacteriophora did not suffer any toxic effect. Of the pathogens tested, the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae were not inhibited in their growth by iridoid glycosides ranging up to 2%. However, an inhibitory effect could be observed against the entomopathogenic bacterium Bacillus thuringiensis, even at the relatively small concentrations that are common in L. melanocephalus. The antibacterial effect might thus be another important selective value of iridoid glycoside sequestration in this species.     

Phenylethanoids, iridoids and a spirostanol saponin from Veronica turrilliana

From the aerial parts of Veronica turrilliana two phenylethanoid glycosides, turrilliosides A and B and a steroidal saponin, turrillianoside were isolated and their structures elucidated as beta-(3,4-dihydroxyphenyl)ethyl-4-O-E-caffeoyl-O-[beta-glucopyranosyl-(1-->4)-alpha-rhamnopyranosyl-(1-->6)]-beta-glucopyranoside, beta-(3,4-dihydroxyphenyl)ethyl-4-O-E-caffeoyl-[6-O-E-feruloyl-beta-glucopyranosyl-(1-->4)-alpha-rhamnopyranosyl-(1-->6)]-beta-glucopyranoside and (23S,25S)-12beta,23-dihydroxyspirost-5-en-3beta-yl O-alpha-rhamnopyranosyl-(1-->4)-beta-glucopyranoside, respectively. Furthermore, eight known glucosides are reported namely, catalpol, catalposide, verproside, amphicoside, isovanilloylcatalpol, aucubin, arbutin, and 6-O-E-caffeoylarbutin, the latter two for the first time in the genus Veronica. The two phenylethanoid glycosides were found to be potent DPPH radical scavengers. All of the tested compounds were inactive against the representative species of fungi and bacteria.     

Iridoid and caffeoyl phenylethanoid glycosides of the endangered carnivorous plant Pinguicula lusitanica L. (Lentibulariaceae)

This work reports for the first time the identification of the major compounds of Pinguicula lusitanica, an endangered carnivorous plant species, using minimal amounts of plant material. A methanol extract was prepared from in vitro cultured plantlets and analyzed by HPLC–SPE–NMR/HPLC–MS. Three iridoid and five caffeoyl phenylethanoid glycosides were identified. These groups of natural compounds were previously reported in the Lentibulariaceae family and have been used as chemotaxonomic markers in related families.     

Iridoid allosides from Viburnum opulus

Arbutin and four novel iridoid glycoside esters, named opulus iridoids I–IV, have been isolated from foliage of Viburnum opulus (Caprifoliaceae). Each opulus iridoid constitutes an inseparable mixture of two compounds, differing by containing either 2-methyl- or 3-methylbutyric acid in ester linkage at the 1-OH-group in an iridoid glycoside. In all glycosides 2′,3′-di-O-acetyl-β-D-allopyranose is linked through a glycosidic bond to C-11 in the iridoid aglycone. The opulus iridoids differ by the degree of acetylation of the aglycone and by the attachment, in III and IV, of a β-D-xylopyranosyl group at C-4 of the allose moiety. The structures have been elucidated by ¹H and ¹³C-NMR spectroscopy and by cleavage of the glycosidic linkage with boron trifluoride etherate in acetic anhydride, yielding the acetates of the cyclized aglycone and of the appropriate mono- or disaccharide. This is the second report of an iridoid attached to a sugar other than glucose and the second time allose has been encountered in higher plants. The systematic position of Viburnum is briefly discussed.     

Further constituents from the bark of Tabebuia impetiginosa

Further study on the constituents from the bark of Tabebuia impetiginosa (Mart. ex DC) Standley afforded twelve compounds, consisting of four iridoid glycosides, one phenylethanoid glycoside, five phenolic glycosides, and one lignan glycoside, along with seven known compounds. The structures of these compounds were determined based on the interpretation of their NMR and MS measurements and by chemical evidence.     

Constituents from the bark of Tabebuia impetiginosa

The bark of Tabebuia impetiginosa afforded nineteen glycosides, consisting of four iridoid glycosides, two lignan glycosides, two isocoumarin glycosides, three phenylethanoid glycosides and eight phenolic glycosides. Their structures were determined using both spectroscopic and chemical methods. Iridoid glycosides, phenylethanoid glycosides and lignan glycosides had ajugol, osmanthuside H and secoisolariciresinol 4-O-beta-D-glucopyranoside as their structural elements, respectively, whereas the aglycone moieties of the isocoumarin glycosides were considered to be (-)-6-hydroxymellein. Phenolic glycosides had 4-methoxyphenol, 2,4-dimethoxyphenol, 3,4-dimethoxyphenol, 3,4,5-trimethoxyphenol and vanillyl 4-hydroxybenzoate as each aglycone moiety. Additionally, the sugar chains of these isocoumarin glycosides and phenolic glycosides were concluded to be beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside as well as those of osmanthuside H and above phenylethanoid glycosides.