PHYTOTHERAPY RESEARCH Phytother. Res.

14, 303322 (2000)

REVIEW ARTICLE

A Review of Antimycobacterial Natural Products

Sandra M. Newton,1* Clara Lau1,2 and Colin W. Wright1
1 2

The School of Pharmacy, University of Bradford, West Yorkshire, UK Department of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong

Tuberculosis is a chronic infectious disease caused by several species of mycobacteria. Due to multidrug resistant strains of mycobacteria and to a high prevalence of tuberculosis in patients who have acquired human immunodeciency syndrome (AIDS), the number of patients infected with the disease is increasing worldwide. Thus there is an urgent need for new effective antimycobacterial agents to replace those currently in use. In this instance, the plant kingdom is undoubtedly a valuable source for new antituberculosis agents. The present review article reports the ndings from an extensive literature search of all plants that have been assessed for antimycobacterial/antitubercular activity over the past 2030 years. An attempt has been made to summarize the information in order to highlight those promising plant species which are worthy of further investigation as leads for drug development. Over 350 plant species from a wide range of families and origins, containing various chemical classes of compounds, have been screened for such activity. A review of the relevant in vitro assays using different species of pathogenic and non-pathogenic mycobacteria is also included. Copyright # 2000 John Wiley & Sons, Ltd.
Keywords: mycobacteria; tuberculosis; plants; antimycobacterial; natural products; traditional medicine.

INTRODUCTION The infectious killer disease, tuberculosis (TB), is the leading cause of death worldwide from a single human pathogen, claiming more adult lives than diseases such as acquired immunodeciency syndrome (AIDS), malaria, diarrhoea, leprosy and all other tropical diseases combined (Zumla and Grange, 1998). The organism usually responsible is the tubercle bacillus, Mycobacterium tuberculosis (MT), discovered by Robert Koch in 1882. However, M. bovis, which infects cattle may also infect man and M. africanum is a cause of TB in West Africa. Furthermore, a number of normally nonpathogenic mycobacteria, especially M. avium, M. intracellulare and M. scrofulaceum, cause opportunistic infectious disease in patients with AIDS (Horne, 1996). Pulmonary TB, the most common type of the disease, is usually acquired by inhalation of the bacillus from an infectious patient and causes irreversible lung destruction. About one third of the worlds population is currently infected with M. tuberculosis; 10% of those infected will develop clinical disease, particularly those who also have the human immunodeciency

* Correspondence to: Dr S. M. Newton, The School of Pharmacy, University of Bradford, West Yorkshire, BD7 1DP, UK.

virus (HIV) infection (Zumla and Grange, 1998). With the discovery of effective antimycobacterial agents (including ethambutol, isoniazid, pyrazinamide, rifampicin and streptomycin) and a reduction in poverty, there was a drastic decline in the number of TB cases, especially in developed nations. However, since the late 1980s, the number of cases of TB throughout the world has been increasing rapidly due partly to the emergence of multidrug resistant M. tuberculosis. According to the World Health Organization (WHO, 1993), it was expected that the annual death rate caused by TB will reach an overwhelming 3.5 million by the year 2000. Thus the TB problem requires urgent attention. Short course anti-TB regimens initially using at least three rstline drugs (including isoniazid, rifampicin and pyrazinamide) are effective. The major problems faced in tuberculosis control are poor infrastructures for diagnosis and drug supply and failure of patients to complete their course of drugs. This is usually due to poor supervision and medical care and, as a result, drug resistance develops. Second-line drugs (e.g. capreomycin, kanamycin, cycloserine, ethionamide) which are often more toxic have to be used in this case. Furthermore, drugs with broader ranges of activity are also required to target emerging pathogens, such as those of the M. avium complex. Hence, there is a great need to search for and develop new, affordable, anti-TB agents. For as long as man can remember, plants particularly, have been used worldwide in traditional medicines for

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S. M. NEWTON ET AL.

the treatment of disease. It is estimated that even today approximately two-thirds to three-quarters of the worlds population rely on medicinal plants as their primary source of medicines (McChesney, 1995). Today, many of the drugs currently used are derived from natural products or have depended upon a natural product for their development and the recent discoveries of the antimalarial artemisinin and the anticancer agent taxol indicate the continuing importance of plant species in drug discovery. However, only a small proportion of plant species have been thoroughly investigated for their medicinal properties (Frame et al., 1998) and undoubtedly there are many novel biologically active compounds to be discovered. During the past two decades, pharmaceutical companies and research scientists have shown an increased interest in phytomedicine. Currently large numbers of species are being screened for pharmacological activities especially those used in traditional/folk medicine. Although plant species have not so far yielded antibacterial compounds of comparable potency to the antibiotics produced by microorganisms, many plant extracts have been tested for activity against microorganisms in the anticipation that highly active compounds will be found. With the urgent need for new anti-TB agents, it is particularly appropriate at this time to review the literature for information on plant species which have been assessed for antimycobacterial activity. The present review attempts to identify those species, which are worthy of further investigation as leads for drug development and to stimulate further work in this important area.

IN VITRO TESTS FOR ANTIMYCOBACTERIAL ACTIVITY Screening plant extracts for antimycobacterial activity is usually carried out using mycobacteria cultured in various types of broth and agar based media. M. tuberculosis has the disadvantages of being slow growing so that tests take several weeks and containment facilities are needed as it is a dangerous pathogen. Many investigators have therefore used non-pathogenic species of mycobacteria such as M. avium, M. intracellulare and M. kansaii, which like M. tuberculosis are slow growing, and other species including M. chelonei, M. fortuitum and M. smegmatis which are faster growing allowing tests to be completed in a few days. Most commonly, the test methods employed are the disc diffusion and the broth dilution methods. In the disc diffusion method, paper discs impregnated with the extract under test are placed on a semi-solid (agar based) medium which has been inoculated with mycobacteria. After incubation, zones of inhibition of bacterial growth around the discs are measured. The main disadvantages with this method are that non-polar compounds may not diffuse into the agar so that active compounds may be missed and that it is not possible to obtain reliable quantitative results for comparative purposes. In the broth dilution method, the minimum concentration required to inhibit bacterial growth (minimum inhibitory concentration, MIC) is determined using a series of tubes containing serial dilutions of the extract in inoculated broth; however solubilization of the extracts
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under test may be a problem (Satim and Washington, 1991). For high-throughput screening, rapid methods which can be automated are needed; these have been reviewed by Gordon et al. (1996). The rst rapid methods developed involved measuring the evolution of 14CO2 from M. tuberculosis cultured in medium containing 14 C-palmitic acid and formed the basis for the BACTEC system (Becton-Dickinson, Oxford, UK). This is used for the susceptibility testing of clinical isolates and can provide results in an average of 5 days compared with 34 weeks for conventional methods. However, the BACTEC system is not suitable for high-throughput screening due to the technical difculties involved in measuring 14CO2. Chung et al. (1995) developed an assay based on measuring the uptake of radiolabelled uracil into M. aurum, a fast growing and non-pathogenic species which appears to be a good model for M. tuberculosis as it has a similar prole of sensitivity to anti-TB drugs. The latter method may be used for highthroughput screening and does not require containment facilities but the separation of incorporated from unincorporated uracil is labour intensive. The major disadvantage of the above is the need for radiolabelled substrates but this has been overcome with the development of assays in which mycobacterial viability is determined using either bacterial or rey luciferase. The bacterial enzyme uses reduced avin (produced by viable mycobacteria) to oxidize an added aldehyde substrate (decanal) which is accompanied by the production of light at 490 nm. Firey luciferase is dependent upon ATP generated by the mycobacteria to decarboxylate luciferin, resulting in the production of light at 562 nm. Light production may be measured easily using a luminometer in high-throughput systems. Several species of mycobacteria, including M. tuberculosis, have been genetically modied by inserting the genes for the production of bacterial luciferase; only viable bacilli emit light when decanal is added and there is no requirement for growth so that susceptibility testing may be carried out rapidly. Similarly, the gene for rey luciferase has been incorporated into a number of mycobacteria species including M. aurum (Chung et al., 1995). Colorimetric methods are very suitable for use in microtitre plates and the results may be easily obtained using a spectrophotometer. Gomez-Flores et al. (1995) reported an assay for testing against the M. avium complex which depends on the ability of viable bacteria to reduce dimethylthiazoldiphenyltetrazolium (MTT) to formazan. Another similar method utilizes the redox dye Alamar blue which changes colour from blue to pink in the presence of viable M. tuberculosis (Yajko et al., 1995). These assays have the advantages of being simple and do not require radioactive substrates but require several days for growth of the bacteria so that they may not be as rapid as the bioluminescence methods described above. While simple antimycobacterial tests are convenient for screening crude plant extracts and isolated compounds, it must not be forgotten that M. tuberculosis is primarily an intracellular pathogen residing in the acidic vacuoles of macrophage cells. This environment may affect the action of anti-TB drugs such as streptomycin (activity reduced) and pyrazinamide (activity increased) and it may be valuable to evaluate the ability of plant
Phytother. Res. 14, 303322 (2000)

Table 1. Medicinal plants/natural products which have been assessed for antimycobacterial activity
Active Constituent(s)/ extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Bromhexine Ambroxol (semi-synthetic derivatives of alkaloid vasicine)

MT

Ethanol extract

High activity

Copyright # 2000 John Wiley & Sons, Ltd.

Inhibitory dilution (no growth or less than 5 colonies) = 1:320 Average MIC for 5 clinical isolates of MT with ambroxol was 64 mg/mL Average MIC for bromhexine was 128 mg/mL for 3 clinical isolates of MT When agent dissolved in DMSO, MIC of bromhexine was lowered Grange and Davey, 1990 Grange and Snell, 1996 (Quassinoids) Both compounds have in vitro inhibitory effects against MT Preparations of the owers, leaves and roots have been widely used in India for the treatment of tuberculosis, asthma and also as expectorants (Dymock et al., 1893). Compounds are widely used as mucolytic agents. Ambroxol has been shown to be well tolerated clinically when given orally in up to 500 mg doses twice daily (Oosterhuis et al., 1993). Activities of all quassinoids screened were very Rahman et al., 1997 low. `19% inhibition at 12.5 mg/mL. Rifampicin was used as the positive drug control Shinjulactone-K Ailanthone Shinjudilactone Diethyl ether extract Guinea-pigs Percentage inhibitions at 12.5 mg/mL are: 19% 17% 15% Inhibitory effect at 0.5 mg/mL and 1.0 mg/mL Jain, 1998 Allicin (diallyl thiolsulnate) isolated from water extract Mean MIC (n = 6) is 1.67 mg/mL for MT Delaha and Garagusi, 1985 Allicin (diallyl thiosulnate) Inhibitory effect of the two higher concentrations of garlic extract0.5 and 1.0 mg/mL. Guinea-pigs, which were inoculated with MT and given garlic extract, produced fewer marked lesions in the viscera. Positive drug controls included, isoniazid, paraaminsalicylic acid and streptomycin All 17 species of MT were inhibited by various concentrations of extract. However, the mean MIC demonstrated very low activity Garlic has been used in traditional Chinese and Egyptian medicine for many centuries (Bolton et al., 1982; Yuang, 1954) Signicant activity. Positive controls included isoniazid, streptomycin, ethambutol, rifampicin. Signicantly active Abbruzzese et al., 1987 Aqueous extract Methanol extract MIC (mean) 1 1.72 mg/mL 2 2.29 mg/mL 3 1.96 mg/mL MIC (complete inhibition) = 1:640 dilution Both parts of the plant completely inhibited growth of MT and MA at 50 mg extract/disc Active. (Concentration not stated) Fitzpatrick, 1954 McCutcheon et al., 1997 Gottshall et al., 1949 Ethanol extract Ethanol extract Alcohol extract Active. (Concentration not stated) No inhibition of MT using a 1:50 dilution Signicant activity. Isoniazid was used as positive control. Previously used in traditional medicine by First Nation's people Activity against MT. Activity present when screened against SA and EC Activity against MT. No activity when screened against SA and EC No signicant activity Gottshall et al., 1949 Wang, 1950

Actaea spicata (Ranunculaceae) Adhatoda vasica (Acanthaceae)

India

Leaf

BCG MT

Ailanthus altissima (Simaroubaceae)

MT

Allium sativum (Liliaceae)

Bulb

MT

Bulb

MT and 16 other species of mycobacteria

ANTIMYCOBACTERIAL NATURAL PRODUCTS

Allium sativum (Liliaceae)

Bulb

Leaf

1 MT 2 MAC 3 MK

MT

Alnus rubra Bong. (Betulaceae)

Br C

Bark Catkin

MT MA

Aloe chinensis (Liliaceae)

Leaf

MT

Leaf

MT

Aloe succotrina (Liliaceae) Alpinia Kumatake Mak. (Zingiberaceae)

China

MT

305

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306

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Amyris elemifera L. (Rutaceae)

Guad

MT MA MK

Taxalin (Oxazole)

Texalin signicantly active

Rastogi et al., 1998

Ethanol extract Inhibitory dilution (No growth or less than 5 colonies) = 1:160 Too toxic in high doses to be clinically useful (Wren, 1988)

Calyx

Ethanol extract

MIC (inhibited more than 99% of the bacterial population) (mg/mL) Method 1 (BACTEC) = 25, 25, 25 Method 2 (agar dilution) = 25, 50, 50 For MT, MA, MK respectively depending on the method used MIC (MT) = 1:80 dilution Signicant activity Used in homeopathic practice for relief of pain and inammation (Wren, 1988) No signicant activity towards the mycobacterium and very low antibacterial activity. Growth controls using Lowenstein-Jensen medium were carried out Signicant activity/moderate toxicity

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Lucas et al., 1951 Grange and Davey, 1990 Fabry et al., 1998 Methanol extract Petroleum ether extract AC = Mulinane Diterpenoid Concentration of 2 mg/mL allowed the growth of more than 10% of the inoculum of the 5 mycobacterial strains. Against the fast growing bacteria MIC = 8 mg/mL MIC (complete growth inhibition) of AC = 20 mg/mL IC50 vs Vero cells was 184 mg/mL Wachter et al., 1998 Ethanol extract MIC (no growth or less than 5 colonies) = 1:640 dilution. Completely inhibited growth of MT at 50 mg extract/disc MIC (MT) = 1:80 dilution Active against MT at 100 mg/mL No activity against MAC, MSi and SLM at 1000 mg/mL MIC 1 8 mg/mL 2 8 mg/mL 3 128 mg/mL Antitubercular activity also demonstrated in extracts obtained from seven other Azorella plants (A. compacta, A. monanthos, A. patagonica, A. lamentosa, A. trifurcata, A. crassipes, A. cryptantha) High activity Grange and Davey, 1990 Signicant activity No inhibition of MA with 50 mg extract/disc. Positive control - isoniazid Signicantly active McCutcheon et al., 1997 Methanol extract Ethanol extract Ethanol extract Lucas et al., 1951 Van Puyvelde et al., 1994 IC50 vs. Vero cells = 1 71.8 mg/mL 2 39.8 mg/mL 3 103.6 mg/mL Weak activity. Used in Rwandese traditional medicine (Van Puyvelde et al., 1975, 1977, 1982) Controls using conventional antituberculosis drugs Signicant activity of both compounds 1 and 2 Highest level of anti-mycobacterial activity found in ower extract Cantrell et al., 1996 1 (24R)- 24,25epoxycycloartane 2 (3aH, 24R)-24,25epoxycycloartane 3 (23R)-3-oxolanosta-8,24dien-23-ol All isolated from dichloromethane extract Dichloromethane extract 100% inhibition against MT at 0.1 mg/mL. 99% inhibition against MA at 1 mg/mL. Signicant activity Positive controls include rifampicin, clarithromycin Cantrell et al., 1998a

Antirrhinum majus (Scrophulariaceae) Arnica montana (Asteraceae)

MT and other bacterial species MT

Azadirachta indica A. Juss. (Meliaceae)

East Africa (Kenya)

Stem/ Bark/ Leaf

MT MA MC MI and bacterial strains

Azorella madreporica Clos (Apiaceae)

Chile

Aerial

MT

S. M. NEWTON ET AL.

MT

Br C

Root

MT MA

Flower

Myroxylon balsamum var. pereirae (Leguminoseae) Balsoamorhiza sagittata Pursh Nutt. (Asteraceae) Barbarea vulgaris (Cruciferae) Bidens pilosa L. (Asteraceae)

Rwanda

Leaf

MT and other bacterial species MT MAC MSi SLM

Borrichia frutescens L. (Sea Daisy) (Asteraceae)

USA

Flower Leaf Stem

MT

Phytother. Res. 14, 303322 (2000)

Borrichia frutescens L. (Asteraceae)

USA

Flower

MT MA

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Brucea antidysenterica (Simaroubaceae)

MT

Inhibition at 12.5 mg/mL 1 15% 2 9% 3 8% 4 3% 5 1% 7% inhibition of MT at 12.5 mg/mL

Activities of quassinoids were very low. 019% inhibition at 12.5 mg/mL Rifampicin was used as the positive control drug

Rahman et al., 1997

Brucea javanica (Simaroubaceae)

MT

1 Dehydrobruceantin 2 Bruceantin 3 Dehydrobruceantarin 4 Bruceanol-F 5 Dehydrobruceantinol (Quassinoids) Bruceoside-D (Quassinoid)

Rahman et al., 1997 Frame et al., 1998

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Rastogi et al., 1998 Rastogi et al., 1998 Ghosal and Chaudhuri, 1975; Ghosal et al., 1978 Herbert et al., 1994 Ingolfsdottir et al., 1998 McCutcheon et al., 1997 Lu et al., 1998

Callistemon citrinus (Myrtaceae)

Puerto Rico

Leaf

Canella winterana (Canellaceae)

Guad

Leaf

Guad

Oil from leaf

Canscora decussata Schult (Gentianaceae)

India

ANTIMYCOBACTERIAL NATURAL PRODUCTS

Centella asiatica Linn. (Umbelliferae)

India

Cetraria islandica L. (Parmeliaceae)

Iceland

Chaenactis douglasi Hook. (Asteraceae)

Br C

Chrysoma pauciosculosa Michx. (Asteraceae)

USA

Root

Activities of quassinoids were very low. 019% inhibition at 12.5 mg/mL Rifampicin was used as the positive control drug LC50 = 168 mg (Brine Signicant activity. Positive (streptomycin) and Ethanol extract Inhibition zones for MSm MSm ranged from 27 mm (1000 mg/ MT negative (untreated) controls were included. shrimp) mL) to 8 mm (25 mg/mL). Mice No toxicity to mice Transitory inhibition of MT similar to growth MT sensitive at 100 mg/mL Artemia salina Leech when given 500 mg/ inhibitory effects for known bacteriostatic (Brine shrimp) agents 100 mL of plant extract/ mouse i.p. over 15 days No signicant activity against MT, MA, MK MIC (inhibited more than 99% of Canellal (Sesquiterpene MT bacterial population) of AC was dialdehyde) isolated from MK greater than 100 mg/mL for each chloroform extract MA test organism No signicant activity MIC (inhibited more than 99% of Essential oil made up of MT myrcene, b- farnesene linalol bacterial population) of essential MK and nerolidiol (all non-cyclic oil was greater than 100 mg/mL MA for each test organism terpenoids) MIC (concentration required for No obvious toxicity Signicant activity of total xanthones and MIC MT Xanthone 1 (Mangiferin) was comparable to that of streptomycin or side effects in growth inhibition) of total Albino rats Xanthone 2 Xanthone 1 showed only weak inhibitory albino rats with xanthones = 10 mg/mL Xanthone 3 activity total xanthones MIC of xanthone 1 alone = 200 Xanthone 4 isolated from Used in traditional Indian medicine to treat (50 mg/kg i.p.) mg/mL ethanol extract tuberculosis (Chopra et al., 1956) No signicant activity on the acid fastness and MT No reduction in the number of viability of MT in vitro. acid-fast bacilli with various Asiaticoside (glycoside) has been isolated concentrations of the active previously from this plant and has been used to extract treat leprosy patients from very early times (Boiteau et al., 1949; Kakkar, 1988) MAu Proto-lichesterinic acid MIC = 250 mg/mL Reputed to be effective in treatment of pulmonary tuberculosis (Vartia, 1973). Controls included rifampicin, streptomycin, isoniazid Signicantly active MT Methanol extract Complete inhibition of MT at MA 50 mg extract/disc. Small zone of clearing of MA at 50 mg extract/disc. Signicantly active MIC (MT)= MT Positive controls are rifampicin against MT and 1 12.5 mg/mL MA 1 (4Z,8Z)-Matricaria ester clarithromycin against MA 2 25 mg/mL 2 (2Z,8Z)-Matricaria ester 3 25 mg/mL 3 (2Z,8-dehydro)-Matricaria MIC (MA)= ester and other related 1 50 mg/mL compounds 2 25 mg/mL 3 25 mg/mL

307

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308

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Ethanol extract Ethanol extract Water extract Usnic acid (dibenzofuran derivative) isolated from diethyl ether extract Water extract Water extract Berberine bisulphate (alkaloid) Methanol extract Methanol extract MIC of active constituent was 32 mg/mL MIC (complete growth inhibition) = 1:640 dilution MIC (no growth or less than 5 colonies) = 1:1280 dilution MIC = 1:80 dilution Signicant activity

MT

Alcoholic extract

MIC = 1:50 dilution

Signicant activity

Wang, 1950 Lucas et al., 1951 Grange and Davey, 1990 Fitzpatrick, 1954 Ingolfsdottir et al., 1998

Leaf Calyx High activity Likely to be too toxic for clinical use (Wren, 1988). Signicantly active

MT and other bacterial species MT

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Activity but lower than the MIC for control drugs, rifampicin (2 mg/mL), streptomycin (0.25 mg/mL) and isoniazid (0.03 mg/mL) Effective in the treatment of pulmonary tuberculosis (Vartia, 1973) Signicant activity Signicant activity Signicant activity but highly toxic MIC (complete inhibition) = 1:640 dilution MIC (complete inhibition) = 1:640 dilution MIC = 1:800 dilution Fitzpatrick, 1954 Fitzpatrick, 1954 Wang, 1950 Berberine highly toxic by parenteral injection (Chang, 1948) Complete inhibition of MT and MA at 50 mg extract/disc Concentration of 2 mg/mL allowed the growth of more than 10% of the inoculum of the ve mycobacterial species Signicant activity. Positive control-isoniazid No signicant activity against the mycobacteria. MIC 50% (mg/mL) ranged from 0.54 depending on bacteria McCutcheon et al., 1997 Fabry et al., 1998 Matricaria lactones Signicant activity of one particular compound (4Z, 8Z-matricaria ester) which gave a MIC of 12.5 mg/mL against MT MIC (MT) 1 825 mg/mL 2 825 mg/mL 3 b25 mg/mL MIC (MSm) 2 0.78 mg/mL Lu et al., 1998 Dichloromethane extract Ethanol extract MIC (mg/mL), for both organisms, ranged from 12.5 b100 for each of the compounds tested 100% inhibition against MT and MA at 0.1 mg/mL Active (concentration not stated) Signicant activity Positive controlsrifampicin, clarithromycin Activity Also active against SA Cantrell et al., 1998a Gottshall et al., 1949 Mitscher and Baker, 1998a 1 Phaseollidin 2 Erythrabyssin II 3 Erygibiso-avone (All avonoids)

Leaf

MT

Chrysanthum sinense China Sab. (Asteraceae) Chrysanthemum segetum (Asteraceae) Cinnamomum camphora (Lauraceae) Cinnamomum zeylanicum (Lauraceae) Iceland Cladonia arbuscula Wallr. Rabenh. (Cladoniaceae)

MAu

Leaf

MT

Leaf

MT

Clematis integrifolia (Ranunculaceae) Clematis virginiana (Ranunculaceae) Coptis coinensia French (Ranunculaceae)

China

Root

MT

S. M. NEWTON ET AL.

Br C

Stem

Empetrum nigrum L. (Empetraceae) Entada abyssinica A. Rich (Leguminosae)

East Africa (Kenya)

Stem

Erigeron philadelphicus L. (Asteraceae)

MT MA MT MA MC MI MTe and 5 bacterial species MT MA

USA

Roots

Leaf

MT MA MT

Erigeron strigosus Muhl. (Asteraceae) Eriodictyon glutinosum (Hydrophyllaceae) Erythrina gibbosa (Papilionaceae)

Phytother. Res. 14, 303322 (2000)

Panama

MT MSm

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Brazil Essential oil Very weak activity

Leaf

Essential oil

Very weak activity

Eucalyptus botryoides Smith (Myrtaceae) Eucalyptus camadulensis Dehn (Myrtaceae)

Brazil

Leaf

MDR MT MT and 8 other species MDR MT MT and 8 other species

Moreira et al., 1997; Leite et al., 1998 Moreira et al., 1997; Leite et al., 1998

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Essential oil Only the slow growing mycobacteria were sensitive using 10mg/mL of essential oil Only the slow growing mycobacteria were sensitive using 10 mg/mL of essential oil. MT, MA and MDR MT were sensitive at 5 mg/mL of essential oil Only slow growing were sensitive at 5mg/mL except MK Very weak activity Moreira et al., 1997; Leite et al., 1998 Essential oil Moreira et al., Very weak activity against MA and MS. No activity against all other species of mycobacteria 1997; Leite et al., 1998 Very weak activity Moreira et al., 1997; Leite et al., 1998 Essential oil MA and MSc were sensitive at 10 mg/mL of essential oil. All other fast and slow growing ones were resistant Only MDR, MT, MT, MSc, MA were sensitive at 10 mg/mL Active (concentration not stated) Only active against all slow growing mycobacteria, except MA and MG, at 5 mg/mL Only active against all the slow growing mycobacteria at 5 mg/ mL, except MK Only active against all the slow growing bacteria at 5 mg/mL, except MK and MM 100% inhibition of MT and MA at 1 mg/mL MIC (mg/mL) of AC = 1.25 for each of the mycobacterial species Very weak activity Very weak activity Ethanol extract Essential oil Gottshall et al., 1949 Moreira et al., 1997; Leite et al., 1998 Very weak activity Moreira et al., 1997; Leite et al., 1998 Very weak activity Moreira et al., 1997; Leite et al., 1998 Activity Cantrell et al., 1998a Al-Yahya et al., 1998 Essential oil Essential oil Dichloromethane extract Ferulenol (Coumarinosesquiterpene) Methanol extract Small zone of clearing of MT at 50 mg extract/disc. No activity against MA at 50 mg extract/disc MIC (no growth or less than 5 colonies) = 1:320 dilution Signicant activity. Ferchromone was another compound identied which had an MIC of 50 mg/mL for each of the mycobacterial species and an MIC of 12.5 mg/mL for each of the bacterial species Used in traditional medicine for skin infections, fever and dysentery Positive controlisoniazid McCutcheon et al., 1997 Ethanol extract High activity Used to treat diarrhoea and fevers Preparation listed in the British pharmaceutical codex, 1934. Grange and Davey, 1990

Eucalyptus citriodora Hook (Myrtaceae)

Brazil

Leaf

Eucalyptus deglupta Smith (Myrtaceae)

Brazil

Leaf

Eucalyptus globulus Labil (Myrtaceae)

Brazil

Leaf

Leaf

MDR MT MT and 8 other species of mycobacteria MDR MT MT and 8 other species of mycobacteria MDR MT MT and 8 other species of mycobacteria MT

Eucalyptus grandis Smith (Myrtaceae)

Brazil

Leaf

Eucalyptus maculata Hook (Myrtaceae)

Brazil

Leaf

Eucalyptus tereticornis Smith (Myrtaceae)

Brazil

Leaf

ANTIMYCOBACTERIAL NATURAL PRODUCTS

USA

Aerial

MDR MT MT and 8 other species of mycobacteria MDR MT MT and 8 other species of mycobacteria MDR MT MT and 8 other species of mycobacteria MT MA

Euthamia leptocephala Greene (Asteraceae) Ferula communis (Umbelliferae)

Saudi Arabia

Rhizome

MI MX MC MSm and other bacterial species

Br C

Leaf

MT MA

Fragaria vesca L. var. bracteata Heller Davis (Rosaceae) Galipea ofcinalis (Angustura vera) (Rutaceae)

MT

309

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Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Galipea ofcinalis (also known as Cusparia febrifuga Humb) (Rutaceae) (Angostura bark)

Bark

MT

Ethanol extract (Alkaloids) 1 Cusparine 2 Galipine 3 4-methoxy-2-npentylquinoleine 4 N-Methyl-2-quinolone

MIC (mg/mL) Ranged from 6.25629 depending on the fraction and the strain of MT

Houghton et al., 1999

Copyright # 2000 John Wiley & Sons, Ltd.

Methanol extract Active, however, none of the compounds showed activity as great as the two positive controls usedisoniazid and rifampicin Traditionally used as a bitter tonic and febrifuge Similar alkaloids from other Galipea species have shown activity against Leishmania, Trypanosoma, Plasmodium species (Spath and Pikl, 1930; Fournet et al., 1993, 1994, 1996) and snails (Vieira and Kubo, 1990) Signicant activity Positive controlisoniazid Signicant activity Positive controlisoniazid McCutcheon et al., 1997 McCutcheon et al., 1997 Methanol extract Complete inhibition of MT at 50 mg extract/disc. No inhibition of MA at 50 mg extract/disc Complete inhibition of MT and MA at 50 mg extract/disc Licoiso-avone (Flavonoid) Ethanol extract Methanol extract Less active against MSm (50 mg/ mL) than MT (25 mg/mL) MIC (no growth or less than 5 Likely to be toxic for High activity colonies) = 1:160 dilution clinical use No signicant activity against Against the fast growing bacteria, the extract the mycobacteria showed MICs and MBCs of !8 mg/mL Mitscher and Baker, 1998a,b Grange and Davey, 1990 Fabry et al., 1998 Methanol extract Ethanol extract Ethanol extract Berberine (alkaloid) MIC of AC MSm = 25 mg/mL BCG = 200 mg/mL MAC = 50 mg/mL Signicant activity. Positive controlisoniazid McCutcheon et al., 1997 Complete inhibition of MT and MA at 50 mg and 10 mg extract/ disc Active (concentration not stated) Gottshall et al., 1949 Gentry et al., 1998 Berberine (alkaloid) Ethanol extract Methanol extract Signicant activity Activity also towards SA and EC Active. Berberine has been demonstrated to reduce the infectivity of bacteria, fungi and protozoa in animals and humans by inhibiting the adherence of microorganisms to the host cells (Amin et al., 1969; Preininger, 1975; Subbaiah and Amin, 1967; Sun et al., 1988) The anti-TB effect was probably due to the large amount of berberine contained in the plant Signicant activity Signicant activity at 10 mg extract/disc towards MA Positive controlisoniazid Signicant activity Positive controlsrifampicin, clarithromycin Mitscher and Baker, 1998a,b More active in vitro against MSm (25 mg/mL) than against MT Active. (concentration not stated) Gottshall et al., 1949 McCutcheon et al., 1997 Cantrell et al., 1998a Small zone of clearing of MT at 50 mg extract/disc. Greatly inhibited growth of MA at 10 mg extract/disc Dichloromethane and hexane 100% inhibition against MT at extracts 0.1 mg/mL

Br C

MT MA

Br C

Root

MT MA

China

MT MSm MT

Geum macrophyllum Willd.var. macrophyllum (Rosaceae) Glehnia littoris F. Schmidt ssp Leiocarpa (Mathias) Hult. (Umbellifereae) Glycyrrhiza glabra L. (Leguminoseae) Guaiacum ofcinale (Zygophyllaceae) Harrisonia abyssinica Oliv. (Simaroubaceae)

East Africa (Kenya)

Root

S. M. NEWTON ET AL.

Heracleum maximum Bartr. (Umbelliferae)

Br C

Root

MT MA MC MTe MI and 6 bacterial species MT MA

Flower

MT

Humulus lupulus (Cannabinaceae) Hydrastis canadensis (Golden Seal) (Ranunculaceae)

Eastern North America

Root

MSm MAC BCG and various bacterial species

North America

Leaf

MSm MT MA MT

Hypericum calycinum (Hypericaceae) Hypericum perforatum L. (Hypericaceae)

Br C

MT MA

Phytother. Res. 14, 303322 (2000)

Inula helenium L. (Asteraceae)

USA

Root

MT

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Root

MT

1 alantolactone 2 iosalantolactone 3 11,13-dihydroisoalantolactone (Eudesmanolides) isolated from hexane, dichloromethane and methanol extracts. Ethanol extract Has purgative properties Signicant activity. Positive control-isoniazid Signicant activity, particularly of compound 1, against all the mycobacterium species and compound 2 against MSm. Controls included streptomycin and isoniazid (MIC = 10 mg/mL) Signicantly active, but may be unsuitable due to purgative property

Signicant activity Rifampicin was used as the positive control Used in traditional medicine for treatment of lung disorders and against TB (Moerman, 1986) Eudesmanolides are also found in Montanoa speciosa and Rudbeckia subtomentosa

Cantrell et al., 1996b

Copyright # 2000 John Wiley & Sons, Ltd.

100 mg/mL crude hexane and chloroform extracts exhibited 100% inhibition of MT. 100 mg/mL methanol extract exhibited 83% inhibition MIC of AC (mg/mL) = 1 32 2 32 3 b128 MIC (no growth or less than 5 colonies) = 1:160. Partial inhibition = 1:320 dilution Grange and Davey, 1990 McCutcheon et al., 1997 Muhammad et al., 1992 Methanol extract Greatly inhibited growth of MT at 50 mg extract/disc. Small zone of clearing of MA at 50 mg extract/disc MIC(mg/mL)= 1 5 mg/mL against each species 2 32 mg/mL (only tested on MSm) 3 Inactive (only tested on MSm) 4 Not tested MIC of compound 1 = 1.25mg/mL for each species of mycobacteria More potent than its abietane derivatives. (MIC = 5.0 mg/mL and 2.5 mg/mL for ()-ferruginol and ()-totarol respectively) Also active against the genera of bacteria Muhammad et Signicantly active but less potent than the al., 1996 positive control, amikacin sulphate (MIC = 0.25 mg/mL against each species.) However, MIC values for streptomycin and isoniazid were higher at 10 mg/mL. Compounds 2 and 3 were not tested against the mycobacteria but compound 2 was weakly active against the bacteria species and compound 3 was inactive Mitscher et al., 1985 Karwinaphthol A inhibited growth of MSm at 12.5 mg/mL and Karwinaphthol B at 50 mg/mL Diameter of the zones of inhibition (in mm) ranged from 2035 depending on the year the samples were harvested and the species tested against. Signicant activity. Seeds are poisonous but fruit pulp is edible. Used locally in Mexico to treat convulsions (Usher, 1974) Signicant antimycobacterial activity on all the species Extracts also were comparable to the standards Standards included Amikacin and Kanamycin Gabbrielli et al., 1988 Ethanol extract followed by partition between n-hexane and acetonitrile Ethanol extraction yielded the compounds, 1 Ferruginol 2 Sandaracopimeric acid 3 Hinokinol 4 3b-hydroxy-sandaracopimeric acid Ethanol extract, which was further partitioned between chloroform and aqueous MeCN. Chloroform fraction yielded diterpenes, 1 7b-Hydroxyabieta-8,13-dien-11,12dione Other abietane derivatives of compound 1 which were tested included ()-ferruginol and ()-totarol 2 Cryptotrienolic acid 3 Isocupressic acid Dichloromethane and ethanol extract. Karwinaphthol A Karwinaphthol B Essential lavandino oils

Ipomoea purga (Jalapa) (Convolvulaceae)

MT

Juniperus communis L. (Cupressaceae)

Br C

MT MA

Juniperus excelsa M.Bieb. (Cupressaceae)

Saudia Arabia

Leaf

MSm MI MX MC

ANTIMYCOBACTERIAL NATURAL PRODUCTS

Juniperus procera Hochst (Cupressaceae)

Saudi Arabia

Bark

MI MX MC MSm and bacteria genera

Karwinskia humboldtiana (Rhamnaceae)

USA

Root

MSm and 6 other types of bacteria

Lavandula angustifolia Mill. (lavender) (Labiatae)

France

Flower

MC MF MK MM MSc

311

Phytother. Res. 14, 303322 (2000)

312

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Leucas volkensii Gurke (Labiatae)

Kenya

Aerial

MT

Signicant activity from all the compounds. (E)- Rajab et al., 1998 phytol, phytanol, (Z)-phytol and the mixture of (E)- and (Z)-phytol were the most active and their activities were in same range as ethambutol (0.953.8 mg/mL)

Br C Signicant activity Signicant activity

Root

China

MT MA MT

Signicant activity. Positive controlisoniazid Signicant activity

McCutcheon et al., 1997 Wang, 1950

Copyright # 2000 John Wiley & Sons, Ltd.

MIC Methanol extract 1 2 mg/mL 1 (E)-phytol 2 2 mg/mL 2 Phytanol 3 2 mg/mL 3 (Z)- phytol 4 Mixture of E)-and (Z)-phytol 4 2 mg/mL 5 64 mg/mL 5 Geraniol 6 8 mg/mL 6 Farnesol Methanol extract Complete inhibition of MT and MA at 50 mg extract/disc MIC (complete inhibition) = 1:50 dilution Ethanol extract MIC = 1:80 dilution Ethanol extract Dichloromethane extract MIC = 1:80 dilution Lucas et al., 1951 Lucas et al., 1951 1 Costunolide 2 Parthenolide 3 1(10)-epoxycostunolide (Germacranolides) Dichloromethane extract 100% inhibition against MT and MA at 0.1 mg/mL MIC (mg/mL) vs MT and MA respectively are 1 32, 128 2 16, 64 3 64, 128 Signicantly active. Positive controlsrifampin, clarithromycin Signicant activity Parthenolide was the most active germacranolide against both MT and MA Other a-methylene-g-lactone-bearing sesquiterpene lactones are moderately active against MT with MICs of 64 mg/mL and below Positive controls include rifampicin and clarithromycin Weak activity Cantrell et al., 1998a Fischer et al., 1998 Cantrell et al., 1998a Signicantly active Bactericidal inhibitory pattern on MT growth comparable to that of streptomycin (used as a positive control) Frame et al., 1998 100% inhibition of MT at 1000 mg/mL. 84%92% inhibition of MA depending on part of plant used 10 mm inhibition zone at 25 mg/ disc for MSm. Activity towards MT at 50 mg LC50 = 224.6 mg/mL (Brine shrimp). No toxicity to mice at 500 mg/100 mL of plant extract mouse given i.p. over 15 days 12 mm inhibiton zone at 1000 mg/ LC50=1000 mg/mL disc for MSm. (Brine shrimp) Active towards MT at 250 mg 7 mm inhibition zone at 50 mg/ disc for MSm. Active towards MT at 100 mg Frame et al., 1998 Frame et al., 1998 Methanol extract 112b hydroxykulactone 2 6b hydroxykulactone 3 Kulonate MIC(mg/mL) 1 16 2 4 3 16 Active. Low degree of toxicity at 500 mg/100 mL of plant extract/mouse given i.p. over for 15 days Streptomycin used as positive control Bacteriostatic activity towards MT at 250 mg/mL LC50 = 1000 mg/mL Signicantly active. Low toxicity. Transitory inhibition of MT growth (Brine shrimp) No toxicity in mice similar to the growth inhibitory effects for known at 500 mg/100 mL of bacteriostatic effects. plant extract mouse Streptomycin used as negative control given i.p. over 15 days Signicantly active. However it is poisonous at higher Used in folk medicine to alleviate pain - tea prepared from bark. dose levels Kulonate previously isolated from Melia (Kokwaro, 1976) azedarach (Chiang and Chang, 1973; Ochi et al., 1977) Cantrell et al., 1999a

Root

Lomatium dissectum Nutt (Umbelliferae) Lonicera japonica Th. (Caprifoliaceae) Lupinus hirsutus (Leguminosae) Lupinus polyphyllus (Leguminosae)

MT and other bacterial species MT and other bacterial species

USA

Leaf Stem Root Bark

Magnolia acuminata (Magnoliaceae) Magnolia grandiora and Magnolia virginiana (Magnoliaceae)

MT MA MT MA

S. M. NEWTON ET AL.

Magnolia grandiora L. (Magnoliaceae)

USA

Flower Fruit Leaf

MT MA

Mammea americana (Guttiferaceae)

Puerto Rico

Leaf

Ethanol extract MSm MT Mice Artemia salina Leach (Brine shrimp)

Mangifera indica (Anacardiaceae)

Puerto Rico

Leaf

Marchantia polymorpha (Marchantiaceae)

Puerto Rico

Leaf

Ethanol extract MSm Mice Artemia salina Leach (Brine shrimp) Ethanol extract MSm MT Mice Artemia salina Leach (Brine shrimp)

Phytother. Res. 14, 303322 (2000)

Melia volkensii Gurke (Meliaceae)

Kenya

Seeds

MT

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Momordica charantia L. (Cucurbitaceae)

Puerto Rico

Leaf

Ethanol extract MSm MT Mice Artemia salina Leach (Brine shrimp) Signicant activity. Positive control-isoniazid Signicant activity

Signicantly active

Frame et al., 1998

Copyright # 2000 John Wiley & Sons, Ltd.

McCutcheon et al., 1997 Gottshall et al., 1949 McCutcheon et al., 1997 Reddi et al., 1986 Signicant activity Positive control-isoniazid All compounds signicantly active against MT Used by rst nations people in traditional medicine for a variety of ailments such as diabetes, rheumatism, tuberculosis, colds, headaches and lung ailments (Turner, 1982; Turner et al., 1990) McCutcheon et al., 1997 Kobaisy et al., 1997 Signicant activity at 50 mg/mL Positive control-isoniazid Signicant activity. Controls included streptomycin, isoniazid and ethambutol 30 mm inhibition zone at 500 mg/ LC50 = 33 mg/mL disc for MSm. Active against MT (shrimp). No at 500 mg toxicity in mice at 500 mg/100 mL of plant extract/mouse given i.p. over 15 days Methanol extract Complete inhibition of MT and MA at 50 mg and 10 mg extract/ disc Ethanol extract, Boiling water Active against MT with both extract extracts (concentration not stated). No activity against EC and SA Methanol extract Complete inhibition of MT at 50 mg extract/disc. Small zone of clearing of MA with 50 mg extract/disc Aqueous Complete inhibition of all strains with a 1:1 dilution of the extract Complete inhibition of MT and MA with 10 mg extract/disc Methanol extract 1 Falcarindol 2 Falcarinol 3 Oplopandiol 4 Active oil 1 (C20H28O4) 5 Active oil 2 (C20H30O4) All polyynes isolated from extraction with methanol followed by dichloromethane Methanol extract, ethanol extract, ether extract Salazinic acid Ethanol extract Ginseng is used in traditional medicine Chang et al., 1979 Showed activity against MT and MSi at 1000 mg/mL No activity against all other species MIC = 1:80 dilution Ingolfsdottir et al., 1998 Van Puyvelde et al., 1994 All active constituents methanol and chloroform extracts were active at a concentration of 10 mg/disc against MT and isoniazid resistant MA Falcarinol and active oil 2 (C20H30O4) completely inhibited growth of MT and isoniazid resistant MA at 20 mg/disc Mycobacterial growth inhibited by 100 mg/mL ether extract and by 500 mg/mL of ethanol and methanol extracts MIC = 250 mg/mL Ethanol extract Weak activity Controls included rifampicin, streptomycin, isoniazid Weak activity. Used in Rwandese traditional medicine for the treatment of pulmonary diseases (Van Puyvelde et al., 1975, 1977, 1982) Controls using conventional anti-tuberculosis drugs Signicant activity Lucas et al., 1951

Moneses uniora L. (Ericaceae)

Br C

Aerial

MT MA

Myrica aspleniora (Myricaceae)

Leaf

MT EC SA

Nuphar lutea L. (Nymphaceae)

Br C

Rhizome

MT MA

Leaf

Br C

Inner bark

MT (two resistant strains) MT MA

ANTIMYCOBACTERIAL NATURAL PRODUCTS

Ocimum sanctum Linn. Mant (Unknown) Oplopanax horridus Smith Miq. (Araliaceae) Oplopanax horridus (Devil's club) (Araliaceae)

North America

Inner bark

MT Isoniazid resistant-MA Other bacterial and fungal species

Panax ginseng (Araliaceae)

Root

MT

Parmelia saxatilis L. (Parmeliaceae)

MAu

Pentas longiora Oliv. (Rubiaceae)

Rwanda

Root

MT MAC MSi SLM

Petasites japonicas (Compositae)

Root

MT

313

Phytother. Res. 14, 303322 (2000)

314

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Picrasma ailanthoides (Simaroubaceae)

MT

Activities of quassinoids were very low ranging from 119% inhibition at 12.5 mg/mL Positive control-rifampicin

Rahman et al., 1997

Copyright # 2000 John Wiley & Sons, Ltd.

1 Nigakihemiacetal 2 Nigakilactone-L 3 Neoquassin 4 Nigakihemiacetal-A 5 Quassin 6 Nigakilactone-H 7 Nigakilactone-E 8 Picrasin-A (Quassinoids) Water extract Signicant activity No signicant activity against MT, MA, MK Signicant activity at 50 mg extract/disc for MT Signicant activity Water extract Pilocarpine (alkaloid) Methanol extract Inhibitions at 12.5 mg/mL are 1 12% 2 9% 3 8% 4 8% 5 7% 6 5% 7 5% 8 4% MIC (complete inhibition) = 1:640 dilution MIC (complete inhibition) = 1:640 dilution MIC (mg/mL) was greater than 100 for each test organism Fitzpatrick, 1954 Fitzpatrick, 1954 Rastogi et al., 1998 McCutcheon et al., 1997 Ethanol extract Methanol extract Methanol extract Small zone of clearing of MT and MA with 50 mg extract/disc Complete inhibition of MT with 50 mg extract/disc. Small zone of clearing of MA with 50 mg extract/disc MIC (no growth or less than 5 colonies) = 1:320 dilution Grange and High specic activity. Used to treat pulmonary and urinary infections and is antibacterial in vitro Davey, 1990 (Wren, 1988) Positive control-isoniazid McCutcheon et al., 1997 Signicant activity of 50 mg extract/disc against MT. Positive control-isoniazid McCutcheon et al., 1997 Grange and Davey, 1990 Ethanol extract Complete inhibition of MT with 50 mg extract/disc. Small zone of clearing of MA with 50 mg extract/disc MIC (no growth or less than 5 colonies) = 1:320 dilution. Partial inhibition of MT at 1:640 MIC (mg/mL) of crude extract = 128 mg/mL for both species MIC (mg/mL) of compounds = 1 64 mg/mL (MT and MA) 2 64 mg/mL (MT) and 128 mg/mL (MA) 3 64 mg/mL (MT) and 128 mg/mL (MA) MIC = 1:80 dilution Appears to be non- High activity toxic. Has been given by inhalation to treat pulmonary infections (Nikulin et al., 1979) Signicant activity of all compounds against MT and compound 1 against MA Controls included rifampicin and clarithromycin Valcic et al., 1999 Methanol extract partitioned between dichloromethane and water Organic phase yielded 17 compounds. Active ones included, 1 Viscidone-14-acetate 2 Coniferyl aldehyde 3 Dihydrobenzofuran ligan aldehyde Ethanol extract Signicantly active Lucas et al., 1951

Leaf

MT

Leaf

MT

Pieris oribunda (Ericaceae) Pieris japonica (Ericaceae) Pilocarpus racemosus (Rutaceae)

Guad

Pinus contorta (Pinaceae)

Br C

MT MK MA MT MA

Piper cubeba (Piperaceae)

Indonesia

MT

Br C

MT MA

S. M. NEWTON ET AL.

Polystitchum munitum Kaulf (Polypodiaceae) Populus tremuloides (Salicaceae)

Br C

MT MA

MT

Propolis

Chile

Resin

MT MA

Phytother. Res. 14, 303322 (2000)

Primula malacoides (Primulaceae)

Flower Peduncle

MT

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Prunus mume (Rosaceae)

USA Hong Kong

Berry

MT

Citric acid Malic acid and various other acids

Citric acid was shown to have specic and signicant activity against MT Used in traditional Chinese medicine (Li ShihChen, 1930; Kariyone and Kimura, 1962)

Roper and Ma., 1968; Ma and Roper, 1968

China Ethanol extract Lochnerin (Indole alkaloid) Ethanol extract Ethanol extract Ethanol extract Methanol extract 1 Allolantolactone 3 2 oxoalloalantolactone isolated from dichloromethane extract Ethanol extract Hypargenin F isolated from acetone extract MIC 1 5.6 mg/mL 2 0.46 mg/mL 3 2.0 mg/mL 4 1.2 mg/mL 5 1.2 mg/mL 6 0.89 mg/mL 7 7.3 mg/mL Active Active MIC (complete inhibition) = 1:50 dilution Small zone of clearing of MT and MA with 50 mg extract/disc MIC (no growth or less than 5 colonies) = 1:160 dilution MIC = 1:80 dilution Has a purgative property Active (concentration not stated) Signicant activity MIC was 100 mg/mL for each test organism Signicant activity

MT

Alcohol extract

Signicant activity

Wang, 1950

Leaf

Citric acid extract inhibited growth of MT on agar, giving a 7.1 mm mean radial width of inhibition. This was highest of all acids tested Bioactivity is rather low when compared with drugs such as INH MIC (complete inhibition) = 1:50 dilution MIC = 1:80 dilution No signicant activity against MT, MA, MK

Copyright # 2000 John Wiley & Sons, Ltd.

Lucas et al., 1951 Rastogi et al., 1998 High specic activity but purgative property Signicant activity Positive controlisoniazid Signicant activity Rifampicin was used as the positive control Compounds previously isolated from Eupatorium quadrangularae (Okunade and Wiemer, 1985) Signicant activity Gottshall et al., 1949 Grange and Davey, 1990 Lucas et al., 1951 McCutcheon et al., 1997 Cantrell et al., 1999b 100 mg/mL dichloromethane extract of root gave a 99% inhibition. MIC (mg/mL) = 1 32 2 128 MIC = 1:80 dilution Activity against MT at 250 mg/mL Lucas et al., 1951 Ulubelen et al., 1988 Active against MT Active also against various genera of bacteria with MIC values (mg/mL) ranging from 62.5125 All signicantly active but Norabietanes 2, 4, 5, and Abietane 2 are most potent Test compounds 17 gave comparable values to standard anti-tubercular agents i.e. rifampicin Salvia species used in traditional medicine and have been associated with antibacterial, antituberculous, and antiphlogistic activities (Lin et al., 1989) Ulubelen et al., 1997 1 Norabietane 1 2 Norabietane 2 3 Norabietane 3 4 Norabietane 4 5 Abietane 1 6 Abietane 2 7 Primarane All isolated from an acetone soluble extract Ethanol extract Ethanol extract Alcohol extract Signicant activity. Also active against SA Signicant activity. Also active against SA Signicant activity Gottshall et al., 1949 Gottshall et al., 1949 Wang, 1950

Guad

Root

MT and other bacterial species MT MA MK MT

MT

Punica granatum L. (Punicaceae) Pyrus atrosanguinea (Rosaceae) Rauwola biauriculata (Apocynaceae) Rheum ofcinale (Polygonaceae) Rhamnus cathartica (Rhamnaceae) Rosa canina (Rosaceae)

MT and other bacterial species

Br C

Leaf Stem Flower

MT MA

Rosa nutkana Presl var. nutkana (Rosaceae) Rudbeckia submentosa (Asteraceae)

Root Leaf Stem Flower

MT

Flower

ANTIMYCOBACTERIAL NATURAL PRODUCTS

Eastern Turkey

Root

MT and other bacterial species MT and bacterial strains

Salix caprea (Salicaceae) Salvia Hypargeia Fisch. Et Mey. (Labiatae) Salvia multicaulis Vahl. (Labiatae)

Turkey

Root

MT

Leaf

MT

Root

MT

Phytother. Res. 14, 303322 (2000)

Salvia ofcinalis (Labiatae) Sanguinaria canadensis (Papaveraceae) Sanguisorba ofcinalis L. (Rosaceae)

China

MT

315

316

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Dehydrocostus lactone Citric acid, Malic acid and various other acids Signicant activity

Root

MT and other bacterial species

Ethanol extract

MIC = 1:80 dilution

Signicant activity

Lucas et al., 1951 Cantrell et al., 1998b Roper and Ma, 1968 Ma and Roper, 1968

Santolina chamaecyparissus (Asteraceae) Saussurea lappa (Asteraceae)

MT MA

Copyright # 2000 John Wiley & Sons, Ltd.

Citric acid was shown to have low activity against MT However the hydrazides (from other acids) produce even greater activity than the parent acids Used in traditional Chinese medicine (Li ShihChen, 1930; Kariyone and Kimura, 1962) Solsodomine A Solsodomine B (Pyrrole alkaloids) MIC (mg/mL = 2 (MT) 16 (MA) Citric acid inhibited growth of MT on agar giving a mean radial width of 7.1 mm. This was the highest of all acids tested. Bio-activity is rather low when compared with drugs such as INH Solsodomine A MIC = 10 mg/mL No cytotoxic activity Solsodomine B had no activity towards Vero cells (dose not stated) on MI Signicant activity of Solsodomine A towards El Sayed et al., 1998 MI. Solsodomines A and B did not show in vitro antimalarial, antifungal or cytotoxic activity (concentrations not stated). Signicant activity Lu et al., 1998 All show no signicant anti-mycobacterial activity Compound 2 also present in Hardwickia pinnata Compound 4 also present in Juniperus pseudosabina Lu et al., 1995 MIC values (mg/mL) for MT and MA ranged from 12.5 to b100 mg/ml MIC b 100 mg/mL against MT and MA for all 6 compounds tested C-10-O-acylated matricaria esters and a dehydromatricaria ester (Diterpenes) 1 Kolavenol 2 Hardwickiic acid 3 Hydroxymanool 4 Monoacetate 5 Diacetate 6 Entabietic acid isolated from the dichloromethane extract Dichloromethane extract Methanol extract 100% inhibition against MT at 0.1 mg/mL. 80% inhibition against MA at 0.1 mg/mL Concentration of 2 mg/mL allowed the growth of more than 10% of the inoculum of the 5 mycobacterial strains MIC = 1 250 mg/mL 2 125 mg/mL Signicantly active Positive controls include rifampicin and clarithromycin No signicant activity against the mycobacteria Cantrell et al., 1998a Fabry et al., 1998 1 Atranorin 2 Lobaric acid Controls included rifampicin, streptomycin, isoniazid Ingolfsdottir et al., 1998

Schizandra chinensis (Schisandraceae)

China

Berry

MT

Solanum sodomaeum L. (Solanaceae)

Libya

Berry

MI

Solidago canadensis L. (Asteraceae)

MT MA

S. M. NEWTON ET AL.

Solidago rugosa Mill. (Asteraceae)

North America

Root Aerial

MT MA

Solidago arguta Ait. (Asteraceae)

USA

Root

MT MA

Spilanthes mauritiana A. Rich. DC. (Asteraceae)

East Africa (Kenya)

Root Flower

Phytother. Res. 14, 303322 (2000)

Stereocaulon alpinum (Stereocaulonaceae)

Iceland

MT MA MC MTe MI and bacterial strains MAu

Table 1. Continued.
Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Strobilanthus cusia (Acanthaceae)

China/ Taiwan

1 MT 2 MSm 3 MAC

Tryptanthrin (indolequinazolinone alkaloid)

MIC 1 1 mg/mL 2 4 mg/mL 3 2 mg/mL (1 and 3 using BACTEC method and 2 using agar dilution method) More potent against MT (1 mg/ mL) and MA (4 mg/mL) than against MSm (6 mg/mL) MIC = 1:80 dilution

Mitscher and Baker, 1998a

Copyright # 2000 John Wiley & Sons, Ltd.

Tryptanthrin (indolequinazolinone alkaloid) Mitscher and Baker, 1998b Ethanol extract Potency of compound in same range as that of antitubercular drugs already in use, i.e. streptomycin and ethambutol Positive controls included various standard antitubercular drugs Active against MDR MT Chinese/Taiwanese medicinal plant. Has folkloric reputation for the topical treatment of athletes foot High signicant activity. Also active against drug sensitive and drug resistant MT and MAC. Compound also isolated from Polygonum tinctorium and Isatis tinctoria Signicant activity Lucas et al., 1951 Frame et al., 1998 Rastogi et al., 1998 1 Ibogaine 2 Voacangine (Indole alkaloids) Ethanol extract Diterpenediol in ethanol extract 30 mm inhibition zone at 500 mg/ LC50 = 93mg/ml Signicant activity disc for MSm. (brine shrimp). No Active against MT at 500 mg toxicity to mice at 500mg/100ml of plant extract/mouse given i.p. over 15 days. Signicant activity by both compounds MIC (mg/mL) 1 50, 100, 50, 2 50, 50, 100 for MT, MA, MK respectively MIC = 1:80 dilution Signicant activity Lucas et al., 1951 Van Puyvelde et al., 1994 Methanol extract Signicant activity of AC against MT, ranging between 25100 mg/ mL, depending on the strain No activity from AE against MAC and SLM at 1000 mg/mL. Concentration of 2 mg/mL allowed the growth of more than 10% of the inoculum of the 5 mycobacterial strains MIC = 1:80 dilution MIC was !100 mg/mL for each test organism and active constituent Used in Rwandese traditional medicine in the treatment of pulmonary diseases (Van Puyvelde et al., 1975, 1977, 1982) Controls using conventional antituberculosis drugs No signicant activity against mycobacteria Fabry et al., 1998 Ethanol extract Isomeranzin Heraclenol (Coumarins) isolated from chloroform extract Signicantly active No signicant activity Lucas et al., 1951 Rastogi et al., 1998

China/ Taiwan

1 MT 2 MA 3 MSm

Syringa vulgaris (Oleaceae) Syzgium jambos (Myrtaceae)

Puerto Rico

Leaf Flower Leaf

MT

Ethanol extract MSm MT Mice Artemia salina Leach (Brine shrimp)

Tabernaemontana citrifolla (Apocynaceae)

Guad

MT MA MK

Leaf

MT

ANTIMYCOBACTERIAL NATURAL PRODUCTS

Taxus canadensis (Taxaceae) Tetradenia riparia Hochst. Codd. (Lamiaceae)

Rwanda

Leaf

MT MAC MSi SLM

Terminalia spinosa Engl. (Combretaceae)

East Africa (Kenya)

Stem

Whole plant

MT MA MC MTe MI and bacterial strains MT

Teucrium chamaedrys (Labiateae) Triphasia trifolia (Rutaceae)

Guad

Leaf

MT MA MK

317

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Copyright # 2000 John Wiley & Sons, Ltd.

Active Constituent(s)/extract(s) Activity Side effects/ toxicity Remarks Reference

Table 1. Continued.
Methanol extract Extract concentrations of 2 mg/ mL allowed the growth of more than 10% of each of the 5 mycobacterial strains MIC (mg/L) 1 25 (MA) 50 (MT) 2 50 (MA) 50 (MT) 3 b100 (MA) b100 (MT) No signicant activity against mycobacteria Fabry et al., 1998

Plant/Natural product

Origin

Part used

Model used/ Route of administration

Ximenia caffra Sond. (Olacaceae)

East Africa (Kenya)

Root

S. M. NEWTON ET AL.

Zingiber ofcinale (Ginger) (Zingiberaceae) 1 10-gingerol 2 8-gingerol 3 6-gingerol isolated from dichloromethane extract

Rhizome

MT MA MC MTe MI and bacterial strains MT MA

10-Gingerol was the most active compound for the inhibition of MA. However this compound was less active against MT

Hiserodt et al., 1998

BCG, Bacillus Calmette Guerin; Br C, British Columbia; DMSO, dimethyl sulphoxide; EC, Escherichia coli; Guad, Guadeloupe; IC50 50% inhibitory concentration (concentration required to inhibit growth of the bacterial population by 50%); INH, isonicotinic acid hydrazide; LC50, 50% lethal concentration (concentration required to kill 50% of the population of organisms); MA, Mycobacterium avium; MAu, Mycobacterium aurum; MAC, Mycobacterium avium complex; MBC, minimum bactericidal concentration; MC, Mycobacterium chelonae/chelonei; MDR, multi-drug resistant; MF, Mycobacterium fortuitum; MG, Mycobacterium gordonae; MI, Mycobacterium intracellulare; MIC, minimum inhibitory concentration; MK, Mycobacterium kansaii; MM, Mycobacterium marinum; MSc, Mycobacterium scrofulaceum; MSi, Mycobacterium simiae; MSm, Mycobacterium smegmatis; MT, Mycobacterium tuberculosis; MTe, Mycobacterium terrae; MX, Mycobacterium xenopi; SA, Staphylococcus aureus; SLM, Simiae-like mycobacterium; TB, tuberculosis.

Phytother. Res. 14, 303322 (2000)

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319

compounds to inhibit M. tuberculosis within cultured human macrophages. This may be carried out using the methodology of Crowle and May (1990).

more reliable than those from studies where controls were not included.

PLANT SPECIES INVESTIGATED FOR ANTIMYCOBACTERIAL ACTIVITY The crude extracts of many plant species, especially those with ethnomedical uses have been assessed for in vitro antimycobacterial properties but relatively few active compounds have been isolated. In many cases, work will have been discontinued because the extracts exhibited little or no activity at the highest concentrations tested. For the purpose of this review, we have selected mostly those species which have been tested for activity against one or more species of mycobacteria and which have been further investigated to determine the nature of the constituents likely to be responsible for the activity. This information, summarized in Table 1, has been compiled mainly from the literature of the last 2030 years. An extensive literature search was carried out using the Science Citation Index of BIDS (Bath Information Data Services), 1981 to date and PubMed (Medline), 1966 to date. The keywords used (in various combinations) in the search were: Plants; Natural; Remedies; Chinese medicine; Traditional; Herbal; Indian medicine; Tuberculosis; Mycobacteria; Antimycobacterial; Antituberculosis; Tubercle bacilli. Furthermore, detailed searches through journals such as Reviews of Aromatic and Medicinal Plants, for papers and articles which are not included in the two databases (BIDS and Medline), were also included. Note that in some of the studies cited such as those reported by Cantrell et al., 1998a; Fitzpatrick, 1954; Grange and Davey, 1990; Lucas et al., 1951, large numbers of plant species were tested but only those that appear to have the most potent activities are included. In addition, a recent article (prepared at the same time as this present review article) by Lall and Meyer (1999) has also reported signicant antimycobacterial activity of some plants which are not incorporated into Table 1. An attempt has been made to summarize all the relevant information available so that species can be assessed for their potential as leads to anti-TB agents. In studies where standard anti-TB drugs have been used as positive controls, this is indicated in the remarks column of the table as the results may be

PLANT SPECIES AS A SOURCE OF NEW ANTIMYCOBACTERIAL AGENTS Compared with microorganisms, plant species have so far proved disappointing as a source of potent antibacterial agents. However, as is well illustrated in Table 1, a number of plant extracts and compounds do have potent antimycobacterial properties. Examples of the species which appear to be among the most active include Allium sativum, Borrichia frutescens, Ferula communis, Heracleum maximum, Karwinskia humboldtiana, Leucas volkensii, Moneses uniora, Oplopanax horridus, Salvia multicaulis and Strobilanthus cusia. In some cases, compounds have been isolated which have antimycobacterial activities comparable to standard anti-TB drugs, for example (E)- and (Z)-phytol and phytanol which were isolated from Leucas volkensii (Rajab et al., 1998). It is hoped that natural products such as the latter may prove to be useful agents for TB treatment or may be lead compounds from which new drugs may be developed. Bromhexine is a semi-synthetic derivative of the alkaloid vasicine, which is found in Adhatoda vasica, an Indian shrub that has long been used in India for the treatment of TB. Although bromhexine and its metabolite ambroxol have been widely used as mucolytics, these compounds exert a pH dependent inhibitory effect against M. tuberculosis in vitro and they are also concentrated in macrophages. It is suggested that they may be useful as adjucts in TB therapy (Grange and Snell, 1996).

CONCLUSION The data compiled in Table 1 reveal that extracts of plant species from a wide range of families have been shown to have signicant in vitro antimycobacterial activities and that a number of active plant-derived compounds belonging to various chemical classes have been isolated. These ndings should stimulate the search for novel natural product leads towards new anti-TB agents.

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