Natural constituents of Ilex species

Journal of Ethnopharmacology, 20 (1987) Elsevier Scientific Publishers Ireland Ltd.

NATURAL

CONSTITUENTS

121

121-144

OF ILEX SPECIES

F. ALIKARIDIS* Organic Chemistry Laboratories, SW7 2AZ (U.K.)

Imperial College of Science and Technology,

London

(Accepted March 30, 1987)

Summary A literature survey on chemical constituents of ZEexspecies is given. General and medicinal uses of the plant are also described. Introduction The genus Zlex (Aquifoliaceae) consists of over 400 species, which grow as trees or shrubs. They have alternate simple leaves, single or clustered small flowers and red black or yellow berries. ZZexrequires a relatively wet and equable climate and has a world-wide distribution except for arid and arctic areas. Major centres of distribution are Central and South America, with 60 species in Brazil alone. Asia is another important centre with 112 species from China, 33 from Japan, 37 from Indochina, 15 from Burma, 17 from the Malay Peninsula, 21 from the Philippines and 45 in the Sumatra-BorneoNew Guinea region, ZIex is reasonably well documented in the fossil records. There are a few Creataceous (about 125 000 000 years ago) and abundant Tertiary (be~nning 65 000 000 years ago) fossil specimens related to it from every geographic region (Martin, 1977). Uses of ZZexspecies The genus ZZexis composed of species of the so called holly plant; Z. aqui(European or English holly), I. opaca (American holly), Z. cornuta (Chinese holly) and I. crenatu (Japanese holly), are among the most well known species of this genus. The use of the holly plant was known many centuries ago. European pagans offered holly twigs to the fairies of the forests and Remans used to exchange branches of holly as gifts during the Saturnalia. The early Christians used holly to decorate their homes during Christmas. Hollies are grown commercially today to be used for decorative purposes because of their coloured berries and shiny green leaves (Grieve, 1982).

folium

*Present address: Department of Biological Chemistry, School of Medicine, University of Athens, Athens 115 27, Greece 0378-87~1/87~$08.75 0 1987 Elsevier Scientific Publishers Ireland Ltd. Published and Printed in Ireland

122

In Brazil and Paraguay one of the most important species of Ilex, I. paraguayensis, or paraguensis, or parcrguariensis is cultivated. Mate, yerba mate or Paraguay tea is a tea-like beverage being prepared almost exclusively from the dried leaves of this species or, in some cases, mixed in a smaller amount with leaves of other Ilex species indigenous to Brazil, Argentina and Paraguay. According to Peckolt (1883), the beverage was known to South American Indians before the invasion of the Spaniards, in contrast to Joyce (1934), who believes that it became known later. In any case, the plant was first cultivated and developed by Jesuit missionaries, to whom the extensive use of mate as a beverage by the South Americans is due (Herzfeld, 1920; Moreau, 1948). The mixed leaves of I. cassine, I. uomitoria and I. dahoon were the basis of another hot drink used by the Indians of the South-Eastern United States (Venable, 1885; Power and Chesnut, 1919). They named it Yaupon or black drink and they used it as a ceremonial to “cleanse” themselves, because of its ability to cause sweat and vomiting. A drink made also of leaves from I. cassine and I. uomitoria was used, in appropriate dilution, as stimulant tea in Southern North America, particularly during the civil war, when the South was blockaded (Lewis and Elvin Lewis, 1977). European and American hollies f&ish white woods that are widely used for marquetry and inlay work and are sometimes dyed to imitate ebony (Metcalfe and Chalk, 1950). On the other hand bird lime, a sticky substance, is made from the bark of some European and Japanese species (Divers and Kawakita, 1888; Iseda et al., 1954), while the occurrence of rubber in the bark of Ilex has also been reported (Fernandez and Iquierdo, 1944; Fernandez, 1947; Paterson-Jones, 1983). Finally, Ito (1908) and Mel1 (1929, 1941) referred to the possibility of obtaining dyes from certain plant species. Natural constituents of Zlex species The various Ilex species contain many classes of chemical These constituents can be classified as follows.

constituents.

Phenols and phenolic acids Thomas and Budzikiewicz (1980b) isolated vanillic and p-hydroxybenzoic acids from the fruits of I, aquifolium, while a group of Chinese scientists isolated hydroquinone (Peking Inst. 1980b; &in et al., 198Oc), homovanillic acid (&in et al., 1980a,c; Peking Inst. 1980b) and 3,4 dihydroxyacetophenone (Peking Inst. 1980a,b; &in et al., 198Oc; Deng, 1981) from the leaves of I. pubescens. The presence of arbutin in the leaves of I. latifolia has also been reported (Miura et al., 1985). Ichikawa et al. (1973) isolated a new bitter principle, named ilesugerin (1) (Fig. I), as well as another compound, named sugeronin, from the leaves of I. sugeroki. The latter compound is a glycoside of the phenolic compound p-hydroxybenzyl benzoate with D-glucose and D-xylose.

123

,CHZOCOPh

e

H

OCHJ

ff0

/

0

\

If

-

OCHj

bH

Oi,

I

2 Glaberide

1 llesugerin

3

ti, II ’

Cl13

h

4 27p-Coumaroxy

3 Neoilexonol

ursolic acid

OH

6 Abbeokutone

5 llex lactone

glucoside

4 R0 CH3

CHZOH

8R=H 7 Pedunculoside

13 R = xylose, ilexolide

I”

A

124

Cl,

3

CHZoif

9

10 R, = CH,,

R, = OCH,

11 R, = CH,,

R, = OH

12 R, = OH, R, = CH,

14 R, = p-D-Glucosyl(1 R, = CH,,

-+ 2)ol-L-arabinose,

ilexside 1 methyl

15 R, = p-D-Glucosyl(1 R, = p-D-glucose,

ester 20 Hainanenside

-2)~L-arabinose, ilexside

II

16 R, = or-L-2’-acetoxyarabinose,

R, = p-D-glucose

18 R, = or-L-arabinose,

R, = p-Dglucose,

Ziyu glycoside

19 R, = ar-L-arabinose,

R, = H, Ziyu glycoside

I

II

21 R=OH 22 R = H, menisdaurin

Fig. 1.

125

Phenyl propanoids

Up to 1935, the presence of tannin in mate had been reported by several researchers. However, Woodard and Cowland (1935) reported that no genuine tannin is present in mate. Instead, the mate contained caffetannin (a pseudotannin) which: when hydrolyzed, gave caffeic acid. The same year Hauschild (1935) isolated from mate a substance which he believed was related to chlorogenic acid. Deulofeu et al. (1943, 19 $4) confirmed that this substance was an acid and mentioned that its hydrolysis gave caffeic acid. Descartes (1953, 1956) isolated two phenolic substances from mate, by means of paper chromatography. One was chlorogenic acid (3-caffeoyl quinic acid) and the other an oxidized condensation product of chlorogenic and probably isochlorogenic acids. Roberts (1956) reported that paper chromatograms of mate extract gave three very strong spots for chlorogenic, neochlorogenic (5-caffeoyl quinic acid) and isochlorogenic acids. Isochlorogenic acid is a mixture of 4,5-dicaffeoyl quinic, 3,4-dicaffeoyl quinic and 3,5-dicaffeoyl quinic acids. (Scarpati and Guiso, 1963; Corse et al., 1965). Ishikura (1975) separated chlorogenic acid, isochlorogenic acid and caffeoylglucose from the methanolic extract of the fruit skin of 11 Ilex species. Hydrolysis of the extracts afforded caffeic acid. The presence of caffeic acid in the amylic alcohol extract of the hydrolysate of I. aquifolium and I. in tegra leaves, had also been reported by Bate-Smith (1962). Recently, 6-methyl-7-hydroxycoumarin (Peking Inst., 1980a), 6,7-dihydroxycoumarin (aesculetin) (Peking Inst., 1980a,b; &in et al., 198Oc), 6-methoxy-7-hydroxycoumarin (scopoletin) (Peking Inst., 1980b; &in et al., 198Oc) and a new compound, named glaberide I (2) (Peking Inst., 1980b; Qin et al. 1980b), have been isolated from the leaves of I. pubescens. The structure of the last compound was determinated on the basis of IR, UV, NMR, MS and X-ray analysis (&in et al., 198Oc; Lin et al., 1982). An thocyanins

(Table

1)

Robinson and Robinson (1931) were the first to investigate the skins of holly berries (1. aquifolium) for the presence of anthocyanins and found, by means of colour and distribution tests, that they contained a pigment which corresponded to a pelargonidin 3-bioside. One year later the same authors reported that the red berries of I. shepherdii contained a pelargonidin 3-pentoseglycoside (Robinson and Robinson, 1932). Hayashi (1942) isolated cyanidin 3-xylosylglucoside, named ilicicyanin, from I. crenata. He also found (Hayashi and Abe, 1953), by means of paper chromatography, that a pelargonidin 3-hexopentoside and cyanidin 3glucoside (chrysanthemin), were present in a ratio 9: 1 in the red fruit of I. geniculata.

126 TABLE

1

ANTHOCYANINS Compound

Ilex

Cyanidin-3-xylosylgiucoside

I. I. I. I. I.

Cyanidin-3-glucoside

Cyanidin-3sophoroside

species

Plant

amelanchier aquifolium buergeri cassine chinensis

Fruit Fruit Fruit Fruit Fruit

I, coriacea I. crenata

Fruit Fruit

I. decidua I. geniculata

Fruit Fruit

I. I. I. I. I. I. I. I. I. I. I. I.

Fruit Fruit Fruit Fruit Fruit Fruit Fruit Fruit Fruit Fruit Fruit Fruit

georgei glabra integra kiusiana latifolia makropoda mitis myrtifolia nipponica opaca paraguariensis pedunculosa

part

I. rotunda I. serrata I. sugeroki

Fruit Fruit Fruit

I. I. I, I.

uomitoria chinensis coriaceae crenata

Fruit Fruit Fruit Fruit

I. I. I. I.

glabra micrococca paraguariensis pedunculosa

Fruit Fruit Fruit Fruit

I. I. I. I. I.

rotunda sugeroki latifolia pedunculosa pu bescens

Fruit Fruit Spr. leaves Spr. leaves Fruit

References Santamour (1973) Ishikura (1971a,c) Ishikura (1975) Santamour (1973) Ishikura (1971c); Santamour (1973) Santamour (1973) Hayashi (1942); Ishikura (1971c), Santamour (1973); Ishikura and Sugahara (1979) Santamour (1973) Ishikura (1975); Ishikura and Sugahara (1979) Santamour (1973) Santamour (1973) Ishikura (1971b,c) Ishikura (1975) Ishikura (1971b,c) Ishikura (1975) Santamour (1973) Santamour (1973) Ishikura (1975) Santamour (1973) Santamour (1973) Ishikura (1971c); Santamour (1973) Ishikura (197 lc) Ishikura (1971b,c) Ishikura (1975); Santamour (1973) Santamour (1973) Ishikura (197 lc) Santamour (1973) Ishikura (1971c); Santamour (1973); Ishikura and Sugahara (1979) Santamour (1973) Ishikura (1975) Santamour (1973) Ishikura (1971c), Santamour (1973) Ishikura (1971c) Ishikura (1975) Yoshitama et al. (1972) Yoshitama et al. (1972) Santamour (1973)

127 TABLE 1 (continued) Compound

Ilex species

Plant part

References

Cyanidin-3rhamnoglucoside Pelargonidin-3-xylosylglucoside

I. pedunculosa I. aquifolium

Spr. leaves Fruit

I. I. I. I,

Fruit Fruit Fruit Fruit

Yoshitama et al. (1972) Ishikura (1971a,c) Santamour (1973) Ishikura (1975) Santamour (1973) Santamour (1973) Ishikura (1975); Ishikura and Sugahara (1979) Santamour (1973) Ishikura (1971b,c); Santamour (1973) Ishikura (197 5) Ishikura (1971b,c); Santamour (1973) Santamour (1973) Ishikura (197 5) Santamour (1973) Santamour (1973) Ishikura (1971b,c); Santamour (1973) Santamour (1973) Santamour (1973) Santamour (1973) Santamour (1973) Santamour (1973) Santamour (1973) Santamour (1973) Santamour (1973)

Perlargonidin-3-glucoside

buergeri ciliospinose cornuta geniculata

I. georgei I. integra

Fruit Fruit

I. kiusiana I. latifolia

Fruit Fruit

I. leucocladia I. nipponica I. perado I. pernyi I serrata

Fruit Fruit Fruit Fruit Fruit

I, I. I. :. I. I. I. I.

Fruit Fruit Fruit Fruit Fruit Fruit Fruit Fruit

verticillata aquifolium corallina cornu ta leucocladia perado serrata verticillata

Ishikura (1971a) separated two kinds of anthocyanins from the red epicarps of I. aquifolium, which were present in a ratio of 1: 1 and identified them, by means of paper chromatographic analysis as pelargonidin 3-xylosylglucoside and cyanidin 3-xylosylglucoside. The same author (Ishikura, 1971b) also reported the occurrence of the above two anthocyanins in three other species of Ilex, but in different ratios: in I. Zatifolia (8: 2); I. integru (8: 2); I. serratu (9: 1). Further investigations on the anthocyanins of the fruit skin of various Ilex species (Ishikura, 1971c) showed that cyanidin 3-xylosylglucoside was commonly found in the genus Ilex. Thus, in I. pedunculosa, I. rotunda, I. crenata and I. chinensis, cyanidin 3-xylosylglucoside was found together with cyanidin 3-monoglucoside in a ratio of 9: 1, 8: 2, 7 : 3 and 6 : 4, respectively. Santamour (1973) reported the presence of pelargonidin 3-glucoside and pelargonidin 3-xylosylglucoside as well as cyanidin 3-glucoside and cyanidin 3-xylosylglucoside in the fruits of various Iiex species. Within the evergreen subgenus Aquifolium, species belonging to the section Lioprinus produced only cyanidin pigments, whereas in those belonging to the

128

section Aquifolium, pelargonidin was the major anthocyanin. Likewise, in the deciduous subgenus Prinos, species of the section Prinoides, contained cyanidin pigments and those of the section Euprinus, had pelargonidin compounds. Thus, contrary to Ishikura’s results, no cyanidin derivatives were found in I. aquifolium, I. integra, I. latifolia and I. w-rata. In addition, Santamour (1973) reported that a petunidin 3-bioside and traces of a petunidin 3-monoside were found in the fruit of I. crenata, while cyanidin 3sophoroside was the only anthocyanin in the fruit of I. pubescens. Ishikura (1975) published a further survey about anthocyanins and other phenolics in the ripe fruits of Ilex species. Cyanidin 3-xylosylglucoside was identified as the common anthocyanin in all the Ilex species examined except I. micrococca, which contained only cyanidin 3-monoglucoside. Pelargonidin 3-xylosylglucoside was found in those Ilex plants with peduncles on the leaf axil of biennial shoots. On the other hand, species bearing a peduncle on the axil formed between stem and annual shoots have only cyanidin 3-monoglucoside and cyanidin 3-xylosylglucoside. However two deciduous plants, I. macropoda and I. micrococca, in spite of belonging to the first group, they lack pelargonidin glycoside. Yoshitama et al. (1972) examined the sprouting leaves of some Ilex species for the presence of anthocyanins. Chrysanthemin was found in I. latifolia, lycoricyanin in I. rotunda and chrysanthemin and keracyanin (cyanidin-3-rhamnoglucoside) were found in I. pedunculosa. Flavonols

and flavones

Quercetin 3-rutinoside (rutin) was found in the leaves of I. aquifolium (Schindler and Herb, 1955), I. paraguariensis (Roberts, 1956), I. integra (Nakaoki and Morita, 1960) and I. hainanensis (Min and &in, 1984). Tkhelidze and Tsiklauri (1975) reported that the total phenols obtained from the leaves of I. colchica corresponded to 4%, on a weight basis. Quercetin 3-glucosylarabinoside, kaempferol 3-glucosylarabinoside and apigenin 5-glucosylrhamnosyl-7arabinoside were isolated and identified using chromatography, acid hydrolysis, colour reactions and spectrophotometry. Bate-Smith (1962) examined, by means of paper chromatography, the amylic alcohol extract of the acidic hydrolysate of I, aquifolium and I. integra leaves for the presence of phenolics. He mentioned the presence of quercetin and kaempferol, in addition to caffeic acid, and the absence of myricetin, delphinidin, ellagic acid, and cyanidin. Ishikura (1975) reported the presence in very low quantities of quercetin and kaempferol in the hydrolysed fruit extract of I. buergeri, I. kiusiana, I. micrococca, I. nipponica and I. sugeroki and the absence of flavonols in I. chinensis, I. macropoda and I. geniculata. Terpenoids

Personne

(Table

2)

(1884) was the first to isolate from I. aquifolium

a triterpene,

129

TABLE

2

TERPENOIDS Compound

Zlex species

Plant part

References

ol-Amyrin

I. aquifolium

Bark

Ursolic acid

I. I. I. I. I. I. I. I. I.

aquifolium aquifolium crenata goshiensis hancedna integra latifolia paraguariensis aquifolium

Leaves Fruit Bark Bark Bark Bark Bark Leaves Leaves

I. I. I. I. I. I,

aquifolium asprella cinerea crenata hanceana latifolia

Fruit Leaves Leaves Leaves Leaves Leaves

I. I. I. I.

latifolia memecylifolia opaca paraguan’ensis

Fruit Leaves Fruit Leaves

I. I. I. I. I. I, I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I.

perado pu bescens rotunda triflora aquifolium crenata hanceana in tegra latifolia latifolia aquifolium corm4 ta macrocarpa opaca pu bescens aquifolium crenata integra buergeri goshiensts comuta integra latifolia

Leaves Leaves Leaves Leaves Leaves Bark Bark Bark Bark Leaves Leaves Leaves Leaves Leaves Leaves Leaves Bark Bark Bark Bark Leaves Bark Leaves

Personne (1884); Jungfieisch and Leroux (1908) Catalan0 et al. (1978) Thomas and Budzikiewicz (1980b) Yagishita (1957b) Yagishita and Nishimura (1961) Yagishita (19 57b) Iseda et al. (1954) Yagishita (1957a) Mendive (1940); Descartes (1944) Nooyen (1920); Fischer and Linser (1930); Schindler and Herb (1955); Catalan0 et ai. (1978) Thomas and Budzikiewicz (1980b) Arthur et al. (1956) Arthur et al. (1956) Nooyen (1920) Arthur et al. (1956) Kariyone and Hashimoto (1949, 1953); Kariyone et al. (1953) Koyama and Kato (1954) Arthur et al. (1956) West et al. (1977) Nooyen (1920); Hauschild (1935); Mendive (1940) Nooyen (1920) Arthur et al. (1956) Arthur et al. (1956) Arthur et al. (1956) Catalan0 et al. (1978) Yagishita (1957b) Yagishita (1957b) Iseda et ai. (1954) Yagishita (1957a) Yamada (1966) Catalan0 et al. (1978) Gau et al. (1983) Arthur et ai. (19 56) West et al. (1977) Arthur et al. (1956) Catalan0 et al. (1978) Yagishita (1957b) Iseda et al. (1954) Yagishita and Nishimura (1961 Yagishita and Nishimura (1961 Gau et al. (1983) Iseda et ai. (1954) Yamada (1966)

p-Amyrin

Oleanolic acid

Baurenol

Neoilexonol Lupeol

130 TABLE

2 (continued)

Compound

Ilex species

Plant part

References

Tataxerol Uvaol

I. latifolia I. aquifolium I. aquifolium L. latifolia I. aquifolium

Leaves Leaves Fruit Leaves Leaves

Yamada Catalan0 Thomas Yamada Catalan0

I. cornu ta I. aquifolium

Leaves Fruit

Latifoloside A Ilexolide A Ilexside I

I. I. I. I. I. I.

Leaves Leaves Leaves Leaves Roots Leaves

Ilexside II

I. cornuta

Leaves

I. Hainanensis I, sugeroki I. aquifolium

Leaves Leaves Fruit

I. cornuta I. cornuta I. cornuta

Leaves Leaves Leaves

Gau et al. (1983) Thomas and Budzikiewicz (1980b); Budzikiewicz and Thomas (1980) Hase et al. (1973) Hase et al. (1973) Hase et al. (1973) Ochi et al. (1975) Zeng et al. (1980, 1984) Nakanishi et al. (1982); Otsuka Pharm Co. (1983) Nakanishi et al. (1982); Otsuka Pharm Co. (1983) Min and &in (1984) Ichikawa et al. (1973) Thomas and Budzikiewicz (1980a,b) Wenjuan et al. (1986) Wenjuan et al. (1986) Wenjuan et al. (1986)

I. cornu ta

Leaves

Wenjuan et al. (1986)

Erythrodiol 3p-Hydroxylup-20(29)en-30al 27-p-Coumaroxyursolic acid Pedunculoside

Hainanenside Abbeokutone Ilex lactone

glucoside

Ziyu-glycoside I Ziyu-glycoside II Pomolic acid 3p-O-2acetoxyarabino 28-0-glucoside X9-Hydroxyoleanolic acid Sp-0-arabino 28-O-glucoside

oldhami pedunculosa rotunda latifolia pu bescens cornuta

(1966) et al. (1978) and Budzikiewicz (1980b) (1966) et al. (1978)

named ilicic alcohol, which was later identified as a-amyrin by Jungfleisch and Leroux (1908). Twelve years later, Nooyen (1920) reported the occurrence of ursolic acid in four Ilex species: I. paraguariensis, I. aquifolium, I. crenata and I. perado. Fischer and Linser (1930), in a detailed procedure for the detection of small quantities of ursolic acid in plants, confirmed the presence of ursolic acid in I. aquifolium. Hauschild (1935) isolated a crystalline compound from mate, which he thought was a sterol, and consequently named it matesterol. Mendive (1940) isolated a-amyrin and ursolic acid from mate. Comparing the chemical constitution, the melting point and the specific rotation of ursolic acid with the descriptive data of matesterol, he proved that the two compounds were identical. Descartes (1944) found cY-amyrin in the unsaponifiable fraction of petroleum ether, benzene, chloroform, ether or acetone extract of mate infusions. Kariyone’s research group (Kariyone and Hashimoto,

131

1949, 1953; Kariyone et al., 1953) found ursolic acid in the leaves and Koyama and Kato (1954) in the fruits of 1. ~atifo~ia,while Schindler and Herb (1955) found the same acid in I, a~uifo~i~m leaves. Arthur et al. (1956) reported that six of the Ilex species of Hong-Kong, contained ursolic acid, but I. macrocarpa contained oleanolic acid and I. pubescens contained both acids. Yagishita’s research group (Iseda et al., 1954; Yagishita, 1957a,b; Yagishita and Nishimura, 1961) isolated 01-and p-amyrin from the bark of some Ilex species. They also reported the isolation of ilexol (baurenol) from I; crenatu (Yagishita, 1957b), ilexol and lupeol from I. integra (Iseda et al., 1954) and a new triterpenoid ketoalcohol, named neoilexonol(3), from I. goshiensis and I. buergeri (Yagishita and Nishimura, 1961), bark. Yamada (1966) mentioned the presence of lupeol, taraxerol, uvaol and @unyrin and the absence of a-amyrin from the leaves of I. latifolia. West et al. (1977) found ursolic acid in the ripe fruits and oleanolic acid in the leaves of 1, opuca. They also reported that saponins appeared to be present in both leaves and fruits. Catalano et al. (1978) isolated a large number of triterpenes from the leaves of I, aquifozium. Uvaol, ursolic acid and oleanolic acid were isolated from the light petroleum extract after successive concentrations. The unsaponifiable material of the residue contained or-amyrin, fl-amyrin, uvaol, ery~rodiol and baurenol; ursolic and oleanolic acids were obtained from the acetone extract. Thomas and Budzikiewicz (1980a,b) isolated cw-amyrin,uvaol, ursolic acid, a new constituent, 27-p-coumaroxy ursolic acid (4) as well as a new bisnormonoterpene, named ilex la&one (5) from the fruits of I, a~~ifo~i~rn. NMR and MS data of the two new compounds were given (Budzikiewicz and Thomas, 1980). Ichikawa et al. (1973) isolated a new bitter principle (6) from the leaves of I. sugeroki. Its structure was confirmed, by hydrolysis with acid and emulsin, as the glucoside of abbeokutone. Hase et al. (1973) reported the isolation of a new bitter principle, named pedun~uloside (7) from the leaves of I. oldhami, I. pedunculosu and I. rotunda. Acid hydrolysis of the pedunculoside gave mainly glucose and the acids of 8 and 9. Ochi et al. (1975) reported the isolation of two new bitter glycosides, name latifoloside A and B, from the leaves of I. a~~ifo~iurn. The acid hydrolysis of latifoloside A afforded three triterpenoid aglycones (10,11,12), whose structures were determined from spectroscopic and chemical evidence. A white amorphous solid compound, named ilexolide A (13), was isolated by Zeng et al. (1980,1984), from the root of I, pubescens and shown to be a triterpene glycoside con~ning ilexolic acid as the aglycone and xylose as the sugar moiety. Nakanishi et al. (1982) isolated two new triterpene glycosides, named ilexside I methyl ester (14), and ilexside II (15), from the leaves of I. cornuta. Their structures have been elucidated by chemical and spectral analysis, notably a novel and efficient application of high resolution ‘H-NMR spectroscopy. Phytochemic~ examination of I. corn~ta leaves, by Gau et al. (1983), resulted in the isolation and identification of lupeol,

132

3/?-hydroxylup-20(29)-en-30~al and oleanolic acid. Further examination of I. cornutu leaves by Wenjuan et al. (1986) led to the isolation of two new triterpene glycosides (16,17) along with two known pomolic acid glycosides (18,19). Finally Min and &in (1984) isolated a new triterpene glucoside, named hainanenside (20), from the leaves of I. hainanensis. The structure was determined by MS, ‘H-NMR 13C-NMR, and chemical evidence. Sterols The occurrence of sterols in mate has been reported by Sanchez (1942). They resembled cholesterol and ergosterol in their chemical behaviour and gave the same colour reactions. Yamada (1966) isolated sitosterol from the ethanol extract of the leaves of I. latifolia. Knights and Smith (1977) mentioned that sterols of male and female flowers and leaves from I. aquifolium were found to be mainly sitosterol with trace amounts of cholesterol, campesterol and stigmasterol. Catalano et al. (1978) reported the occurrence of p-sitosterol, campesterol, stigmasterol and ergosterol in the leaves of I. aquifolium and Thomas and Budzikiewicz (1980b) found p-sitosterol in the fruits of the plant. Tahara and Sakuda (1980) examined the sterol composition of the seeds of I. latifolia, I. in tegra, I. crenata and I. rotunda. They found p-sitosterol to be the main component with small amounts of cholesterol, campesterol and stigmasterol. Finally, Gau et al. (1983) isolated and identified P-sitosterol-fl-D-glucoside from the leaves of I. cornutu. Purine alkaloids Stenhouse in 1843, was the first to report the presence of caffeine in mate (Palet, 1917). This was confirmed by Peckolt (1883), while Lendner (1918) reported the presence of caffeine in the seeds of I. paraguariensis. Marangoni (1945) compared 10 known methods for the determination of caffeine in mate and reported that the method of Cortes (1939) was the most satisfactory. Power and Chesnut (1919) examined the leaves of American Ilex species and reported that in contrast to I. uomitoria, no caffeine was found in I. cassine, I. glabra, I. leavigata, I. myrifolia and I. opaca. The absence of caffeine from the leaves of I. dumosa had also been reported (Lendner, 1913). Oehrli (1927) and Hauschild (1935) reported the occurrence of theobromine, in addition to caffeine, in mate. This was confirmed by Diaz (1944) and Descartes (1956b, 1958, 1962). Michl and Haberler (1954) studied mate, using electrophoresis and paper chromatography, and found caffeine, theobromine, theophylline and adenine present. The presence of theophylline in addition to caffeine and theobromine was confirmed by Fleischer (1956) and Siesto (1959a,b), while Maravalhas (1965) found caffeine, theophylline and theobromine in the fruits of I. paraguariensis.

133

The presence of theobromine in various Ilex species was confirmed by Bohinc and coworkers, using techniques involving the destination of eutectic melting point and chromatography. They found theobromine in I. aquifolium (Bohinc, 1959, 1967), I. crenatu (Bohinc and Sobocanpazin, 1970), I. caroliunu (Bohinc et al., 1972a), I. cassine (Bohinc et al., 1972b), I. perudo (Bohinc et al., 1975) and 1. umbiguu (Bohinc et al., 1977). In a further survey for the presence of caffeine and theobromine in the leaves of Ilex species they also mentioned the absence of purine alkaloids from the leaves of I. decidua, I. opaca, I. coriuceu, I, glubra, I. pernyi and I. integra (Bohinc and Korbar-Smid, 1978). F&p et al. (1983) examined I. urgent&a for the presence of purine alkaloids and reported that caffeine and ~eophylline were not detected, but theobromine was present in both the stems and leaves of the plant. Amino acids Knapp and Liskens (1954) reported the presence of aspartic acid, cysteine, glutamic acid, serine, tryptophan and valine in the leaves of I. aquifolium and Cascon (1955) the presence of alanine, arginine, asparagine, aspartic acid, cysteine, cyst&e, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, me~ionine, threonine, ~yptoph~, tyrosine and valine in the leaves of 1. paruguuriensis, Schneider (1965) examined the changes in free amino acids in the leaves of I, crenuta Nigru. He detected arginine, hydroxyproline, isoleucine, leucine, lysine, phenylalanine, proline, threonine, tyrosine, valine, alanine, asparagine, aspartie acid, glutamic acid, glycine, histidine and y-aminobutyric acid. However, only the last mentioned eight compounds underwent quantitative changes during hardening off and storage at low temperature. ~ieell~eo~s

nitrogen eom~oun~

Trigonelline (Ribeiro, 1940; Correia, 1947; Descartes, 1962) as well as choline (Descartes, 1956b, 1958,1962; Barreto and Ruy, 1956) have been found in mate. Miura and Tsung-Ming (1984) reported the presence of choline and acetylcholine in the leaves of 1. opacu. Ueda et al (1983) isolated a new cyanoglucoside (21) from the fruits of I. wurburgii and elucidated its structure by chemical and spectral means. A related compound, menisdaurin (22) was also isolated. Futty acids stable 3) The fatty acid composition of the seed oil of I, paruguun’ensis (Cattaneo et al., 1952), 1. integru (Koyama and Toyama, 1957a,b; Hirose et al., 1971; Tahara and Sakuda, 1980), I, macropodu and 1. seratu (Koyama and Toyama, 1957a,b), I. IatifoEia (Kashimoto and Noda, 1958; Tahara and Sakuda, 1980)

134 TABLE

3

FATTY

ACIDS

Compound

Ilex species

Plant part

References --

Laurie

Myristic

Pentadecanoic

Palmitic

Heptadecanoic

Stearic

Nonadecanoic Arachidic

I. I. I. I. I. I. I. I.

crenata integra latifolia paraguariensis rotunda crenata in tegra Eatifolia

Seed Seed Seed Seed Seed Seed Seed Seed

I, I. I. I. I. I. I, I. I. I. I.

pedunculosa rotunda aquifolium crenata in tegra latifolia rotunda aquifolium crena ta in tegra la tifolia

Seed Seed Leaves Seed Seed Seed Seed Leaves Seed Seed Seed

I. I. I. I. I. I.

paraguariensis pedunculosa rotunda aquifolium crenata in tegra

Seed Seed Seed Leaves Seed Seed

I. I. I. I. I. I. I, I.

la tifolia rotunda aquifolium crenata crenata integra la tifolia latifolia

Seed Seed Leaves Bark Seed Seed Bark Seed

I. I. I. I. I. I, I.

paraguariensis pedunculosa rotunda in tegra aquifolium crenata integi-a

Seed Seed Seed Seed Leaves Seed Seed

I. latifolia

Seed

Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Cattaneo et al. (1952) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Kashimoto and Noda (1958); Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Catalan0 et al. (1978) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Crombie (1958); Catalan0 et al. (1978) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Kashimoto and Noda (1958); Tahara and Sakuda (1980) Cattaneo et al. (1952) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Catalan0 et al. (1978) Tahara and Sakuda (1980) Hirose et al. (1971); Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Crombie (1958); Catalans et al. (1978) Yagishita (19 57 b) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Yagishita (1957a) Kashimoto and Noda (1958); Tahara and Sakuda (1980) Cattaneo et al. (1952) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Hirose et al. (1971) Catalan0 et al. (1978) Tahara and Sakuda (1980) Hirose et al. (1971); Tahara and Sakuda (1980) Kashimoto and Noda (1958); Tahara and Sakuda (1980)

135 TABLE 3 (continued)

Compound

Behenic Lignoceric Cerotic Pentadecenoic Palmitoleic

Oleic

Nonadecenoic

Gadoleic Linoleic

Ilex species

Plant part

References

I. I. I. I. I. I. I. I. I. I. I. I. I, I. I.

Seed Seed Seed Leaves Leaves Seed Leaves Leaves Seed Seed Seed Seed Leaves Seed Seed

Cattaneo et al. (1952) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Catalan0 et al. (1978) Catalan0 et al. (1978) Kashimoto and Noda (1958) Catalan0 et al. (1978) Crombie (1958); Catalan0 et al. (1978) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Cattaneo et al. (1952) Tahara and Sakuda (1980) Crombie (1958); Catalan0 et al. (1978) Tahara and Sakuda (1980) Koyama and Toyama (1957 a,b); Tahara and Sakuda (1980) Kashimoto and Noda (1958); Tahara and Sakuda (1980) Koyama and Toyama (1957a,b) Cattaneo et al. (1952) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Koyama and Toyama (1957a,b) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Crombie (1958); Catalan0 et al. (1978) Crombie (1958); Catalan0 et al. (1978) Tahara and Sakuda (1980) Koyama and Toyama (1957a,b); Tahara and Sakuda (1980); Hirose et al. (1971) Kashimoto and Noda (1958); Tahara and Sakuda (1980) Koyama and Toyama (1957a,b) Cattaneo et al. (1952) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Koyama and Toyama (1957a,b) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Crombie (1958); Catalan0 et al. (1978) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980) Tahara and Sakuda (1980)

paraguarieneis pedunculosa rotunda aquifolium aquifolium latifolia aquifolium aquifolium crenata integra paraquariensis rotunda aquifolium crenata integra

I. latifolia

Seed

I. I. I. I. I. I. I. I. I. I. I. I.

Seed Seed Seed Seed Seed Seed Seed Seed Leaves Leaves Seed Seed

macropoda paraguariensis pedunculosa rotunda serrata crenata integra latifolia aquifolium aquifolium crenata integra

I. la tifolia

Nonadecadienoic Eicosadienoic Linolenic

I. I. I. I. I. I. I. I. I.

macropoda paraguariensis pedunculosa rotunda serrata integra latifolia latifolia aquifolium

Seed -Seed Seed Seed Seed Seed Seed Seed Leaves

I. I. I. I.

crenata integra latifolia rotunda

Seed Seed Seed Seed

136

as well as that of I. crenata, I.rotunda and I. pedunculosa (Tahara and Sakuda, 1980), has been reported. Crombie (1958) and Catalano et al. (1978) reported the fatty acid composition of I. aquifoZiu~ leaves while ageing and seasonal effects were reported by Niemann and Baas (1985a, b). Holloway and Deas (1973) mentioned the presence of epoxyoctadecanoic acids in cutins and suberins of I. aquifolium and Yagishita (1957a,b) of stearic acid in the bark of I, ~atifo~~uand I, emnate. Alkanes and alcohols Catalano et al. (1978) isolated alkanes with 26-33 carbon atoms from the unsaponifiable fraction of the light petroleum extract of I. aquifo~iu~ leaves. GLC analysis showed that alkanes with 29 and 31 carbon atoms were the most abundant constituents. West et al. (1977) also reported that nonacosane was the most abundant constituent of a hexane extract of I. opaca leaves. Mellisyl alcohol and a trace of glycerol have been isolated from the cuticle of the leaves of 1. latifolia (Kariyone and Hashimoto, 1953; Kariyone et al., 1953). Thomas and Budzikiewicz (1980b) isolated 2-trichlorometbylpropan-2-01 from the fruits of 1. uquifoliu~ and &in et al. (1980a,c), vomifolio1 from the leaves of I. pu~sce~. Carbohydrates Hauschild (1935) reported the isolation of an inactive inositol from mat& Plouvier (1948) examined the leaves of I. aquifolium, I. cassine, I. crenatu and I. pernyi for the presence of sugar alcohols and sucrose. He reported the absence of quebrachitol and dulcitol and the presence of sucrose in the examined plants. Sucrose, raffinose, glucose and levulose were identified in I. p~ru~urie~sis (Chl~ta~, 1955) and D-fructose, D-galactose, a-D-glucose, fl-D-glUCOSe and sucrose in I. opaca (Fretz et al., 1970; Fretz, 1971; Balge et al., 1978), leaves. D-Glucose has also been isolated as a free sugar from the leaves of I. latifolia (Kariyone and Hashimoto, 1953; Kariyone et al., 1953; Kato et al., 1979). Finally, Chrelashvili and Mg~oblishvili (1974) reported that monosaccharides and sucrose predominated in the young leaves of 1. aquifolium, whereas the maltose fraction increased with age. Vitamins and carotenoids Escudero et al. (1936, 1941) and Mendive (1938) were the first to report the presence of ascorbic acid (vitamin C) in the leaves of I. parcguariensis. Villela (1938, 1939) reported that thiamine (vitamin B,) occurs in the leaves of the above plant. This was confirmed by Escudero et al. (1944, 1945) and Chaves (1944) who also reported the presence of riboflavin (vitamin B2), nicotinic acid and carotene in yerba mate. The results of Sigueira et al. (1953) supported the presence of the above vitamins in the plant.

137

VaIadon et al. (1974) examined and reported the quantitative distribution of carotenoids in I. aquifoliurn berries. They found that phytofluene, (Ycarotene, pcarotene, 5,6 : monoepoxy-P-carotene, 5,6: 5’,6’-diepoxy-Pcarotene and aurochrome are always present and in some cases associated with {carotene, mutachrome, 5,6 : monoepoxylutein, lutein and neoxanthin. Kulikov and Ivantsova (1976) reported that the amount of carotenoids depends on the age and size of the plant leaves. Medicinal actions of Zlex species The progress in the elucidation of the chemical constituents of Zlex species offers a possible insight into the probable chemical basis of the traditional uses of these plants. I. pubescens a traditional Chinese medicinal plant, is used mainly for the treatment of coronary heart diseases (Lewis and Elvin-Lewis, 1977). Phenolics and phenyl propanoids isolated from I. pubescens leaves have been shown to possess antipyretic, anti-inflammatory, analgesic, cardiovascular and circulatory activities (Peking Institute 1980a,b). Koo (1986) proposed that the vasodilator and hyperaemic effects of I. pubescens are correlated with four substituted catechols, which are present in the plant. Zhang et al. (1981) reported that flavonoid glycosides from the bark of the plant are effective in the treatment of hypertension, hyperlipaemia and hepatitis, while a triterpene glycoside from its root exhibits cardiac activity (Zeng et al., 1984). I. cornuta is also known in China as a traditional medicinal plant, used for the treatment of dizziness and hypertension (Wenjuan et al., 1986). They suggested that adenosine must be the active principle of the plant, which is responsible for the increase in coronary blood flow. On the other hand, the leaves of the plant have been reported to contain triterpenoid glycosides with antithrombotic and anticholesteraemic action (Otsuca Pharmaceutical Company, 1983). Glucosides have also been isolated from the bark of the Chinese plant Chin-Pi-Ying (Zlex), which has been popularly used for curing malarial fever (Chu et al., 1956). I. aquifolium leaves have been traditionally used in intermittent fevers and rheumatisms, for their antipyretic properties, as well as an astringent, diuretic and expectorant (Grieve, 1982). I. opaca leaves have been used as a diuretic, tonic, purgative and cardiac stimulant (Grieve, 1982; Lewis and Elvin-Lewis, 1977). Waud (1931,1932) reported that a dried powder emulsion resulting from leaves and berries of I. aquifolium and I. opaca has the pharmacological action of digitalis. Watanabe (1938) on the other hand reported that injection of an extract of I. integra bark into rabbits, causes a definite fall in blood pressure. The depurative, stimulant and diuretic actions for which I. paraguariensis leaves have been traditionally used (Hegnauer, 1964) could be ascribed to their high purine content (Baltassat et al., 1984). Graham (1984) reported that mate constitutes the primary source of methylxanthines in the diet

138

of some groups in South America. In colonial times, a significant number of people in South America used to live on an almost exclusively meat diet and no vitamin deficiency symptoms had been reported (Meyer, 1937; Escudero and Landabure, 1945). According to the authors, the explanation could be found in that adequate vitamins were taken from mate, which was used as a beverage. References Arthur, H.R., Lee, C.M. and Ma, C.N. (1956)

Occurrence of triterpenes in Aquifoliaceae and Ericaceae of Hong Kong. Journal of the Chemical Society, 1461-1463. Balge, R.C., Dunham, C.W., Liebhardt, W.C. and Svec, L.V. (1978) Carbohydrate fractions of American holly, Ilex opaca Ait leaves. Journal of the American Society for Horticultural Science 103, 759-761. Baltassat, F., Darbour, N. and Ferry, S. (1984) Study on the purine levels in caffeine drugs. Plantes Medicinales et Phytotherapie 18, 195-203. Barreto, R. and Ruy, C. (1956) Microbiological determination of choline in herba mate. Reuista Quimica Znd. (Rio de Janeiro) 25 (288), 12-19. Bate-Smith, EC. (1962) The phenolic constituents of plants and their taxonomic significance I. Dicotyledons. Journal of the Linnean Society (Botany) 58 (371), 95-173. Bohinc, P. (1959) Contribution to the knowledge of the chemism of Holly (Zlex aquifolium). Farmacevtski Vestnik 10, 57-58. Bohinc, P. (1967) Determination of theobromine in Zlex aquifolium. Farmaceutski Vestnik

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Bohinc, P., Korbar-Smid, J. and Marinsek, A. (1977) Xanthine alcaloids in Zlex ambigua leaves. Farmacevtski Vestnik 28, 89-96. Budzikiewicz, H. and Thomas, H. (1980) Constituents of Celastrales: Part V 27-p-cumaroxyursolic acid - a new constituent of Rex aquifolium L. Zeitschrift fiir Naturforschung B 35,226-232. Cascon, C.S. (1955) Amino acids in Ilexparaguariensis. Argentina (Rio de Janeiro) 38, 7-l 5.

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Catalano, S., Marsili, A., Morelli, I., Pistelli, L. and Scartoni, V (1978) Constituents of the leaves of Ilex aquifolium L. Planta Medica 33, 416-417. Cattaneo, P., De Sutton, K.G. and Rodrigez, M.L. (1952) Chemical composition of the seed oil of Ilex paraguariensis. Anales Direc. National Quimica (Buenos Aires) 5 (9), 9-12. Chaves, J.M. (1944) Vitamines B,,B, and C in different varieties of yerba mate. Reuista alimentar (Rio de Janeiro) 8 (II), 5-7. Chen, C.T. and Wang, Y.S. (1976) A study on the constituents of Zlex asprella champ. Bulletin of the Institute of Chemistry AcademiaSinica 23, 13-15. Chlamtac, E.B. (1955) Sugars in Ilex paraguariensis. Boletin del Institute de Quimica Argentina

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