МОНОГРАФИИ ВОЗ Т 4

Lichen Islandicus

fumarprotocetraric acid, protolichestearic acid and cetraric acids (5, 13). A biologically active aliphatic -lactone, (+)-protolichesterinic acid has also been reported (14). The structures of fumarprotocetraric acid, cetraric acid and (+)-protolichesterinic acid are presented below.

Medicinal uses

Uses supported by clinical data

Used orally as a demulcent to treat inflammation and dryness of the pharyngeal mucosa (15, 16).

Uses described in pharmacopoeias and well established documents

Used orally for treatment of loss of appetite and nausea (3, 17).

Uses described in traditional medicine

Used to treat asthma, cramps, bronchitis, cough, diabetes, exhaustion, gastric disturbances, immune depletion, migraine, nausea in pregnancy and wounds. Also used as an emergency food source, an emollient and a galactagogue (4, 18–20).

Pharmacology

Experimental pharmacology

Antimicrobial and antiviral activities

An aqueous extract of the thalli and protolichesterinic acid were screened for in vitro activity against Mycobacterium aurum, a nonpathogenic organism with a similar sensitivity profile to M. tuberculosis. Protolichesterinic acid had a minimum inhibitory concentration (MIC) of θ 125.0 μg/ml (21). An aqueous extract of the thalli was screened for in vitro activity against Helicobacter pylori. (+)-Protolichesterinic acid was identified as

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an active component. The MIC range of protolichesterinic acid, in free as well as salt form, was 16.0–64.0 μg/ml (14).

Protolichesterinic acid, isolated from the thalli, inhibited the DNA polymerase activity of human immunodeficiency virus-1 reverse transcriptase, with a median inhibitory concentration (IC50) of 24.0 μM (22).

Antioxidant activity

The antioxidant activity, reducing power, superoxide anion radical scavenging and free radical scavenging activities of an aqueous extract of the thalli were investigated in vitro. The antioxidant activity increased with increasing amount of extract (from 50.0 to 500.0 μg) added to linoleic acid emulsion. About 50.0, 100.0, 250.0 and 500.0 μg of aqueous extract of the thalli exhibited higher antioxidant activity than 500.0 μg of Α-tocopherol. The samples showed 96, 99, 100 and 100% inhibition of peroxidation of linoleic acid, respectively. On the other hand, 500.0 μg of Α-tocopherol showed 77% inhibition of peroxidation of linoleic acid emulsion (23).

Antiproliferative effects

The effects of protolichesterinic acid, isolated from the thalli, on three malignant cell-lines (T-47D and ZR-75-1 from breast carcinomas and K-562 from erythroleukaemia), as well as normal skin fibroblasts and peripheral blood lymphocytes were assessed in vitro. Protolichesterinic acid caused a significant reduction in DNA synthesis, as measured by thymidine uptake, in all three malignant cell lines. The IC50 was between 1.1 and 24.6 μg/ml for protolichesterinic acid. The breast cancer cell-lines were more sensitive than K-562. The proliferative response of mitogen-stimulated lymphocytes was inhibited with an IC50 of 24.5 μg/ml for protolichesterinic acid. These concentrations are of the same order of magnitude as the MICs in the 5- lipoxygenase assay. Significant cell death (assessed by the 3-(4,5-dimethyl- thiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetra- zolium assay and trypan blue exclusion) occurred in the three malignant cell-lines at concentrations of protolichesterinic acid greater than 30.0 μg/ ml. In K-562, morphological changes consistent with apoptosis were detected. At a concentration of protolichesterinic acid of 20 μg/ml, up to 38% cell death was observed in mitogen-stimulated lymphocytes, but unstimulated lymphocytes were clearly less sensitive. In contrast, the DNA synthesis, proliferation and survival of normal skin fibroblasts were unaffected at doses of protolichesterinic acid of up to 20 μg/ml (24).

Immune effects

A polysaccharide, isolated from the lichen, and described as a branched galactomannan, with a backbone composed of two structural elements;

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(1λ6)-linked Α-D-mannopyranosyl and Α-D-(1λ6)-galactopyranosyl units, had pronounced immunostimulating activity. The polysaccharide enhanced granulocytic phagocytosis by 68%, when administered at a concentration of 1 mg/ml, in human granulocytes in vitro. Intraperitoneal administration of the polysaccharide to mice, at a dose of 1.0 mg/kg body weight (bw), increased the rate of carbon clearance, indicating significant activation of the reticuloendothelial system (13, 25, 26). A polysaccharide named Ci-3, isolated from an aqueous extract of the thalli, as well as from an alkali extract of the thalli had immune stimulating activity in vitro. At a concentration of 100.0 μg/ml, the polysaccharide Ci-3 stimulated granulocytic phagocytosis and reduced the complement-induced haemolysis in sheep erythrocytes by 80% (27).

Toxicity

Lichens are known to naturally contain bitter and potentially toxic lichen acids, as well as concentrated heavy metals that are taken up from the environment. The toxicity of the thalli was investigated using traditional preparation methods (boiling, ash-soaking). Untreated and briefly boiled thalli were lethal to mice when added to feed at concentrations of 25 and 50% w/w, but when the thalli were prepared by ash-soaking, the mice tolerated the crude drug reasonably well for 3 weeks (28).

In another study, the toxicity of the traditionally prepared thalli was assessed in mice and rats. As a 50% w/w mixture in normal feed neither the pretreated nor the untreated thalli additive was well tolerated by mice. However, rats tolerated 25% w/w ash-treated thalli in normal feed over 3 months, although the body weight did not increase as much as in control animals. At the end of the experiment, the rats in the thalli-treated group had proteinuria, and on autopsy some tubular changes were found, probably due to high concentrations of lead in the thalli and in the kidneys (29).

Clinical pharmacology

In an open clinical trial, 100 patients (children and adults) suffering from inflammatory irritative conditions of the respiratory tract were treated with lozenges containing an aqueous extract of the thalli. The illnesses ranged from pharyngitis and laryngitis to chronic or acute bronchitis. The patients were treated with lozenges containing 160.0 mg of an aqueous extract of the crude drug, for between 4 days and 3 weeks. After evaluation of the symptoms such as cough, hoarseness, secretion, inflammation and pain, a positive outcome was observed in 86% of patients. The lozenges were well tolerated, with few side-effects and good gastric tolerance (16).

Sixty-one patients who had recently undergone surgery to the nasal septum were included in a randomized, placebo-controlled, clinical trial.

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These patients suffer especially from dryness and inflammation of the oral cavity due to breathing only through the mouth while the nose is permanently closed by a nasal package. Three groups of patients treated with three different concentrations of an aqueous extract of the thalli (0.048 g drug per pastille/day, 0.3 g drug per pastille/day or 0.5 g drug per pastille/day) were compared. Coating, dryness and inflammation of the mucosa, lymph nodes and tongue, the patients’ tolerance of the drug, and symptoms such as hoarseness and sore throat were documented. The oral application was performed from the first to the fifth day after the operation. The treatment with the lichen caused a direct reduction of the observed pathological changes. There were no significant differences between the three test groups. A dose of 0.48 g per day (10 pastilles per day) is a sufficient treatment (15).

Adverse reactions

Allergic reactions, abdominal pain and nausea have been reported. Few adverse events were observed in a study of 3143 children aged 4–12 years who were administered 320–600 mg of an aqueous extract of the thalli in lozenges per day over a 1–2 week period. The adverse events were mild and transient (30).

Contraindications

Hypersensitivity or allergic reactions to the crude drug.

Warnings

No information was found.

Precautions

Carcinogenesis, mutagenesis, impairment of fertility

No information was found.

Other precautions

No information was found.

Dosage forms

Crude drug, extracts, fluidextract, infusion and tinctures (3, 17).

Storage

Store in tightly sealed containers away from light and moisture (5).

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Posology

(Unless otherwise indicated) (3, 17, 18)

As a demulcent to treat inflammation and dryness of the pharyngeal mucosa: comminuted herb for infusions and other Galenical formulations for internal use. For loss of appetite: comminuted herb, cold macerates for internal use.

Average daily dosage: 4–6 g of herb in divided doses, or equivalent preparations (17). Infusion: 1.5 g of crude drug in 150 ml water. Fluidextract 1:1 (g/ml): 4–6 ml. Tincture 1:5 (g/ml): 20–30 ml (18).

References

1.European Pharmacopoeia, 5th ed. Strasbourg, Directorate for the Quality of Medicines of the Council of Europe (EDQM), 2005.

2.Bedevian AK. Illustrated polyglottic dictionary of plant names. Cairo, Medbouly Library, 1994.

3.British herbal pharmacopoeia. Exeter, British Herbal Medicine Association, 1996.

4.Farnsworth NR, ed. NAPRALERT database. Chicago, University of Illinois at Chicago, IL (an online database available directly through the University of Illinois at Chicago or through the Scientific and Technical Network [STN] of Chemical Abstracts Services) 30 June 2005.

5.Wichtl M. (Bisset NG, ed and trans.) Herbal drugs and phytopharmaceuticals, English ed. Boca Raton, FL, CRC Press, 1994.

6.Evans WC. Trease and Evans Pharmacognosy, 15th ed. Edinburgh, WB Saunders, 2002.

7.Gathercoal EN, Wirth EH. Pharmacognosy, 2nd ed. Philadelphia, Lea & Febiger, 1948.

8.Gudjónsdottir GA, Ingólfsdottir K. Quantitative determination of protolichesterinicand fumarprotocetraric acids in Cetraria islandica by high-performance liquid chromatography. Journal of Chromatography A, 1997, 757:303–306.

9.WHO guidelines on assessing quality of herbal medicines with reference to contaminants and residues. Geneva, World Health Organization, 2007.

10.Deutsches Arzneibuch 10th ed. (DAB 10). Stuttgart, Deutscher Apotheker Verlag, 1991.

11.Guidelines for predicting dietary intake of pesticide residues, 2nd rev. ed. Geneva, World Health Organization, 1997 (WHO/FSF/FOS/97.7).

12.Kramer P et al. Rational approach to fractionation, isolation, and characterization of polysaccharides from the lichen Cetraria islandica. Arzneimittelforschung, 1995, 45:726–731.

13.Ingolfsdottir K. Bioactive compounds from Iceland moss. Proceedings of the Phytochemistry Society of Europe, 2000, 44:25–36.

14.Ingolfsdottir K et al. In vitro susceptibility of Helicobacter pylori to protolichesterinic acid from the lichen Cetraria islandica. Antimicrobial Agents Chemotherapy, 1997, 41:215–217.

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15.Kempe C et al. Isländisch-Moos-Pastillen zur Prophylaxe bzw. Heilung von und ausgetrockneter Rachenschleimhaut [Icelandic moss lozenges in the prevention or treatment of oral mucosa irritation and dried out throat mucosa]. Laryngorhinootologie, 1997, 76:186–188 [in German].

16.Vorberg G. Erfahrungsbericht: Flechtenwirkstoffe lindern Reizzustände der Atemwege: Noben den entzündungshemmenden Eigenschaften wirkt sich der Schleimhautschutz besonders günstig aus. Ärztliche Praxis, 1981, 85:3068.

17.Blumenthal M et al. The complete German Commission E monographs: therapeutic guide to herbal medicines. Austin, TX, American Botanical Council, 1998.

18.Blumenthal M et al, eds. Herbal medicine: expanded Commission E monographs. Newton, MA, Integrative Medicine Communications, 2000.

19.Lesiovskaya EE et al, eds. Pharmacotherapy with the principles of phytotherapy, Moscow, Geotar-Med Publishers, 2003 [in Russian].

20.Mommsen H. [Icelandic moss as a drug.] Zeitschrift für Allgemeinmedizin, 1970, 46:1566–1567 [in German].

21.Ingolfsdottir K, Jurcic K, Wagner H. Immunomodulating polysaccharides from aqueous extracts of Cetraria islandica (Iceland moss). Phytomedicine, 1998, 5:333–339.

22.Pengsuparp T et al. Mechanistic evaluation of new plant-derived compounds that inhibit HIV-1 reverse transcriptase. Journal of Natural Products, 1995, 58:1024–1031.

23.Gulcin I et al. Determination of antioxidant activity of lichen Cetraria islandica (L) Ach. Journal of Ethnopharmacology, 2002, 79:325–329.

24.Ogmundsdottir HM et al. Anti-proliferative effects of lichen-derived inhibitors of 5-lipoxygenase on malignant cell-lines and mitogen-stimulated lymphocytes. Journal of Pharmacy and Pharmacology, 1998, 50:107–115.

25.Ingolfsdottir K et al. Immunologically active polysaccharide from Cetraria islandica. Planta Medica, 1994, 60:527–531.

26.Ingolfsdottir K et al. Antimycobacterial activity of lichen metabolites in vitro. European Journal of Pharmaceutical Sciences, 1998, 6:141–144.

27.Olafsdottir ES et al. Immunologically active (1 λ 3)-(1 λ 4)-alpha-D-glucan from Cetraria islandica. Phytomedicine, 1999, 6:33–39.

28.Airaksinen MM, Peura P, Antere S. Toxicity of Iceland lichen and reindeer lichen. Archives of Toxicology (Suppl) 1986, 9:406–409.

29.Airaksinen MM et al. Toxicity of plant material used as emergency food during famines in Finland. Journal of Ethnopharmacology, 1986, 18:273–296.

30.Hecker M, Volp A. Tolerability of Icelandic moss lozenges in upper respiratory tract diseases – multicentric drug monitoring study with 3,143 children.

Forschende Komplementarmedizin und klassische Naturheilkunde, 2004, 11:76–82.

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Fructus Macrocarponii

Definition

Fructus Macrocarponii consists of the fresh or dried ripe fruit of Vaccinium macrocarpon Ait. (Ericaceae) (1).

Note: Cranberry liquid preparation is official in the United States Pharmacopeia (2).

Synonyms

Oxycoccus macrocarpus (Alt.) Pursh., O. painstris var. macrocarpus (1).

Selected vernacular names

Afonya, American cranberry, bear berry, black cranberry, cranberry, Kronsbeere, large cranberry, low cranberry, marsh apple, Moosbeere, Preisselbeere, suureviljaline jõhvikas, trailing swamp cranberry (1, 3, 4).

Geographical distribution

Native to eastern North America (1).

Description

Trailing evergreen shrub up to 15 cm in height; rhizomatous. Stem: slender, glabrous to hairy, rooting at nodes. Leaf: simple, alternate, sub-sessile; blade narrowly elliptic, rarely oblong, (5–) 7–10 (–18) mm long, (2–) 3–4 (–5) mm wide, apex rounded; leathery, upper surface green, lower surface glaucous; margin entire, rarely revolute. Inflorescence: flowers solitary in leaf axils of new shoots; bracts 2, green, 1–2 mm wide; pedicels 2–3 cm, recurved, jointed to the flower. Flower: perfect, radially symmetrical with a nectariferous disc surrounding the style; calyx 4-lobed; corolla white to pink, cup-shaped, 4-lobed with lobes much longer than cup and strongly reflexed at anthesis; stamens 8, filaments with stiff hairs along margins, anthers dehiscent by apical pores; ovary inferior, style 1, stigma capitate. Fruit: berry, globose, 9–20 mm in diameter, glabrous, red to crimson, dark burgundy or almost black, many-seeded. Chromosome number: n = 12 (1).

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Plant material of interest: dried ripe fruit

General appearance

The dried berry is dark red to almost black, 7.0–11.5 mm in width and 10–15 mm in length, with a smooth but deeply wrinkled, slightly lustrous surface. At the fruit apex is the dried, slightly raised nectariferous disc, 1.5–2 mm across, with a shallow depression in its centre inside which is a small protuberance from the remains of the style. At the fruit base is a small scar where the berry was attached to the pedicel. The texture is spongy. In cross-section, the mesocarp and endocarp are a dull red. The mesocarp has large air pockets, and varies in thickness when dried from 0.1–2 mm. In cross-section, the fruit is divided into 4 chambers (carpels), each running the entire length of the fruit. The chambers are separated by dull red, thin, translucent septa and each chamber contains 1–5 seeds. Each seed is narrowly ovoid with an acute apex, 1–2.7 mm long and approximately 1 mm wide, with a lustrous rose to red testa that is longitudinally wrinkled. The endosperm is an opaque white (1).

Organoleptic properties

Odour: slightly fruity and sweet; taste: very acidic and tart (1).

Microscopic characteristics

Fruit: the exocarp consists of anthocyanin-coloured polygonal cells covered by a thick cuticle. Groups of cells are separated by fairly thick, colourless walls, whereas the walls within the groups are very thin. The mesocarp consists of large, spherical, thin-walled cells in which small bundles of spirally thickened vessels are embedded.

Seed (cross-section): the epidermis of the testa is composed of radially elongated rectangular cells filled with mucilage. The walls are thickened in a U-shape, the thickest wall being the exterior one; the mucilage is radially striated, and in the centre of the cell is a small lumen. Below the epidermis are several layers of polygonal cells with thick, brown, occasionally reticulately thickened walls. These cells are 250–350 Μm long and approximately 80 Μm broad. The innermost layer of the testa consists of compressed, rectangular cells with sinuous longitudinal walls. The voluminous endosperm is made up of small polygonal and oil-containing cells (1).

Powdered plant material

Numerous fragments of the exocarp with colourless cell walls and violet contents; thin-walled parenchyma of the mesocarp; thickened and pitted cells of the testa; oil droplets (1).

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General identity tests

Macroscopic and microscopic examinations and high-performance thinlayer chromatography (1).

Purity tests

Microbiological

Tests for specific microorganisms and microbial contamination limits are as described in the WHO guidelines on assessing quality of herbal medicines with reference to contaminants and residues (5).

Foreign organic matter

To be established in accordance with national requirements.

Total ash

To be established in accordance with national requirements.

Acid-insoluble ash

To be established in accordance with national requirements.

Water-soluble extractive

To be established in accordance with national requirements.

Alcohol-soluble extractive

To be established in accordance with national requirements.

Loss on drying

To be established in accordance with national requirements.

Pesticide residues

The recommended maximum limit of aldrin and dieldrin is not more than 0.05 mg/kg (6). For other pesticides, see the European pharmacopoeia (6) and the WHO guidelines on assessing quality of herbal medicines with reference to contaminants and residues (5) and pesticide residues (7).

Heavy metals

For maximum limits and analysis of heavy metals, consult the WHO guidelines on assessing quality of herbal medicines with reference to contaminants and residues (5).

Radioactive residues

Where applicable, consult the WHO guidelines on assessing quality of herbal medicines with reference to contaminants and residues (5).

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Other purity tests

Chemical tests to be established in accordance with national requirements.

Chemical assays

To be established in accordance with national requirements.

Major chemical constituents

Contains organic acids (e.g. citric and quinic acids predominate), phenolic acids (e.g. chlorogenic and p-coumaric acids). The fruits contain up to 10% tannins (mostly catechol tannins). The major flavonoids are myricetin and quercitin glycoside. The primary constituents of interest in cranberry are the anthocyanins and related compounds based on the anthocyanidins, cyanidin and peonidin, to which are attached a glucose, arabinose or galactose sugar at the number 3 carbon, giving a total of 6 anthocyanins. The galactosides of peonidin and cyanidin predominate, with moderate amounts of the arabinosides. Also of interest are proanthocyanidins (condensed tannins) consisting of oligomers of two flavan-3-ols, primarily epicatechin plus small amounts of epigallocatechin. Cranberry proanthocyanidins contain a variety of complex linkages, including A-type, B-type and branching chains. A-type proanthocyanidins have doubly-linked flavan-3-ol units with both a carbon–oxygen–carbon (C2–O–C7) bond and a carbon–car- bon (C4–C8 or C4–C6) bond (1, 3). Structures of cyanidin, peonidin, myricetin and proanthocyanidin trimer are presented below.

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