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Unan; S,S'•• 01"
I.I- -lul"n
Shakir Jamil*, Shoaib Ahmad**, Jamal Akhtar and Khursheed Alam
Dept. of Moalijat, Facultyof Unani Medicine, lamia Hamdard, New Delhi-l10062, India
* Correspondent author, E-mail: [email protected]
**1437, Sector 39 - B, Chandigarh-160036, India
Introduction
Diseases caused by protozoa are
responsible for a considerable morbidity
and mortality,especially in the developing
world. Amoebic dysentery or amoebiasis
is one of such protozoal diseases. It is
caused by Entamoeba histolytica. If
untreated, it may give rise to serious
complications including liver abscesses.
There are some 42 million cases annuallyand an estimated 75,000 deaths across the
world due to this disease. In allopathic
systemofmedicine the disease is generallytreated with metronidazole, although this
drug is poorly tolerated by some patients.
There are a number of antiamoebic drugsherbal origin mentioned in the classical
texts in Unani System of Medicine
(Table 1). Some of these drugs havebeneficial effects in treating the disease inthe experimental animals (Table 2) andhumans in the clinical situations
(Table 3). Both in vivo and in vitro
methods have been developed to assay the
antiamoebic activity of the herbal drugs(single botanicals as well as multicomponent formulations).
In in vivo antiamoebic assays
rats are used for determination of activity
against intestinal infectionswhile hamsters
are used for evaluating activity against
hepatic infections. E. histolytica isintroduced into the caecum via rectum
and the. intestine is examined for the
presence of amoebae and ulceration. Liver
infections are initiated by injection ofamoebae into the 10bes.However,in vivo
tests are difficult to perform, timeconsuming and are unpleasant for theanimals.
For in vitro antiamoebic assaysthe development of axenic cultures of
Entamoeba histolytica has enabled the
development of in vitro assays.Before the
development of axenic media, it was only
possible to grow the amoebae in presenceof bacteria (polyxenic culture) and it
made interpretation of the results
extremely difficult. Now-a-days, E.histolytica is grown in 96-well
microculture plates in the presence of
serial dilution of herbal drug extracts orisolated compounds. Atthe end of the test
time, the growth of amoebae may beevaluated by visual observation with a
microscope. Alternatively,a colorimetricmethod may be used which involves the
removal of culture medium thus, leavinghealthy amoebae attached to the bottom
of wells while the dead amoebae are
washed away. A measure of the number
of amoebae remaining in the well is
obtained by fixing and staining the
parasites. The quantity of stain taken up isproportional to the number of amoebae
and can be determined byspectrophotometric method.
A relatively simpler method i.e.,
micro dilution technique for the
assessment of in vitro activity againstEntamoeba histolytica has been
developed and validated withmetronidazole. This test has been used to
detect the antiamoebic activitiesofextracts
of Brucea javanica (Linn.) Merrillfruits and Quassia amara Linn. stems.
The activity was found to be associated
with quassinoid-containing fractions. The
50% inhibitory concentrations for somequassinoids against amoebae were
determined by using this microdilution
method. These concentrations rangedfrom 0.019 j.lglmlfor bruceantin, the most
active quassinoid, to greater than 5 j.lgl
ml for glaucarubol, the least active
compound tested. The micro dilution
technique, being more accurate and
precise will be quite useful in searchingthe novel antiamoebic drugs.
Natural Product Radiance Vol 2(1) January-February 2003
Artid• 0-~ ~ -_ e .. ,_ _- _> .,.>--".)1"--, .. -_".7-_"./",-r_"./
Table 1: Antiamoebic Drugs in Unani System of Medicine
Botanical name
Acacia arabica
Acacia catechu
Aegle marmelosAlstonia scholaris
Balanites aegyptiacaBauhinia racemosa
Boswellia glabraCalotropis proceraCannabis sativa
Cassia fistulaCinnamomum camphoraCinnamomum zeylanicumCochlospermum gossypiumCordia latifoliaCydonia oblongaEmblica officinalisFicus bengalensisFicus carica
Ficus glome rataHelicteres isora
Holarrhena antidysenterica
Hyoscyamus nigerMalva rotundifoliaMalva sylvestrisMangifera indicaMentha arvensis
Mimosa pudicaMimusops elengiMyrtus communisPapaver somniferumPlantago ovataPlantago majorPunica granatumQuercus infectoriaRosa damascena
Rubia cordifoliaRUinex vasicarius
Syzygium cuminii
Family
Mimosaceae
Mimosaceae
Rutaceae
Apocynaceae
Zygophyllaceae
CaesalpiniaceaeBurseraceae
AsclepiadaceaeCannabinaceae
CaesalpiniaceaeLauraceae
Lauraceae
Cochlospermaceae
BoraginaceaeRosaceae
EuphorbiaceaeMoraceae
Moraceae
Moraceae
Sterculiaceae
Apocynaceae
Solanaceae
Malvaceae
Malvaceae
Anacardiaceae
Lamiaceae
Mimosaceae
Sapotaceae
Myrtaceae
Papaveraceae
Plantaginaceae
PlantaginaceaePunicaceae
FagaceaeRosaceae
Rubiaceae
polygonaceae
Myrtaceae
UnaniName
Gondkikar
Kattha
BelgiriSatoona
HingotKachnar
Lohban
PostAkh
BhangAmaltas
Kafoor
Da1chini
Katira
SapistanBihi
Amla
Bargad
Roghan anjeerGular
Maror phaliKurchi
Ajwain khurasaniKhubbazi
Resha khutmi
Khasta-e-Amba
Podina
LajwantiSamar mulsari
Habbulas
Post Khaskhas
Isapghol
BartangPostAnar
Mazu
GuIqand
MajeethChuka
Jamun
Natural Product Radiance Vol 2(1) January-February 2003 '" .1'. 11
Table 2: Pharmacologically proven antiamoebic Unani Drugs
Botanical name Family Unani name Part used
---------.-------------.-- ..----_-!_.----.-.-.-.--.------Aconitum heterophyllum
Aegle marmelos
Ailanthus glandulosa
Allium sativum
Alstonia angustifolia
Artemisia absinthium
Calotropis procera
Cassia fistula
Cinchona ledgeriana
Commiphora wightii
Cyperus scariosus
Euphorbia hirta
Gossypium herbaceum
Holarrhena antidysenterica
Helicteres isora
Momordica dioica
Myristica fragrans
Piper longum
Pistacia intergerrima
Plantago major
Psidium guajava
Punica granatum
Tabernaemontana spp.
Tylophora indica
Ranunculaceae
Rutaceae
Meliaceae
Liliaceae
Apocynaceae
Asteraceae
Asclepiadaceae
Caesalpiniaceae
Rubiaceae
Burseraceae
Cyperaceae
Euphorbiaceae
Malvaceae
Apocynaceae
Sterculiaceae
Cucurbitaceae
Myristicaceae
Piperaceae
Anacardiaceae
Plantaginaceae
Myrtaceae
Punicaceae
Apocynaceae
Asclepiadaceae
Atis Root
Bel
Fruit
Bakayan
Gall
Lehsun
Cloves
Satoona
Root
Afsanteen
Whole plant
Madar
Root, bark
Amaltas
Whole seed
Quinine
Bark
Muqil
Gum resin
Nagarmotha
Fruit
Doodhi kalan
Whole plant
Banola
Seed
Indrajau talkh
Fruit
Marore Phali
Pod
Jungli Karela
Fruit
Jaiphal
Seed
Filfil Draz
Fruit
Kakra singhi
Gall
Bartang
Seed
"Amrood
Fruit
Rum man
Flower, Rind
Gulchandi
Root
Antamul
Root
Table 3: Clinically proven antiamoebic Unani Drugs
Botanical name FamilyUnani namePart used
---_ •....-._--~._.•._-----_._._._._ .._._. __ ._.-.._""-"'~.•_-'""Artemisia absinthium
AsteraceaeAfsanteenWhole Plant
Calotropis procera
AsclepiadaceaeMadarRoot bark
Helicteres isora
SterculiaceaeMarore PhaliPod
Natural Product Radiance Vol 2(1) January-February 2003
Natural Product Radiance Vol 2(1) January-February 2003
Cephaelis ipecacuanha
Holarrhena antidysenterica
Marketed antiamoebic Unani
preparations
A number of drug houses are
engaged in preparation of Unani
medicines in India. Prominent amongthem are Hamdard Dawakhana (Delhi),
Sadar Dawakhana (Delhi) , Shama
~:::"~-r.",.~ ....."",..... =:.,~-", -",T-",officinale Rose. was tested against
E. histolytica in vitro and in vivo. In
the traditional systemof medicine in India,
the formulation is been prescribed forintestinal disorders. The formulation had
a Minimum Inhibitory Concentration
(MIC) of 1000 ~g1ml as compared with
10 ~g1ml for metronidazole.
~--_1t'Ftllle""",_m?~_-",F' -'"
active against E. histolytica in vitro.Quassinoids possessing an unsaturated A
ring, a lactone ring and a methylene
oxygen bridge in the C-ring such as inbrusatol, a constituent of Brucea
javanica (Linn. ) Merrill , have very
potent action against E. histolytica.Unfortunately,these compounds also have
very high toxicity to mammalian cells andattempts to improve their selectivity by
making structural changes have not so far
been very fruitful. Again, the antiamoebic
activityis due to the inhibition of proteinsynthesis and as these processes appearto be similar in amoebae and mammalian
cells, it is likely to be quite difficult toimprove selectivity. However, it is of
interest to note that one quassinoid
glaucarubinone found in Simaroubaglauca, was formerly used in France for
the treatment of amoebic dysentery.The antiamoebic effectof a crude
drug formulation which comprisedextracts of five well known Unani
medicinal herbs, namely, Boerhaaviadiffusa Linn., Berberis aristata DC.,
Tinospora cordifolia (Willd.) Miersex Hook. f. & Thoms., Terminalia
chebula Retz. and Zingiber
Antiamoebic compounds fromUnani Drugs
Although a large number of
natural products have been shown to be
able to inhibit the growth of amoebae, very
fewhave been shown to be selectivelytoxic
to the parasite.
Berberine, a benzylisoquinolinealkaloid common in members of the
Menispermaceae, has been clinicallyusedin the treatment of leishmeniasis.
Berberine has been reported to be
effectiveagainst E. histolytica in vitro.The plant flavonoids{ (-) -epicatechin,
(- ) -epigallocatechin and kaempferol}
have been found to higWyactive againstamoebae.
Conessine is one of the steroidal
alkaloids from the bark ofHolarrhena
antidysenterica (Linn.) Wall. and has
also shown an in vitro activityagainst E.histolytica. Emetine from Cephaelisipecacuanha (Brot.) A. Rich.(Rubiaceae) has been found to be higWyactive in the treatment of both hepatic andintestinal amoebiasis but has some toxic
effects especially on the heart. It inhibits
protein synthesis which is probably
responsible for its antiamoebic action andthe toxic effects seen in man. Several
alkaloids from Strychnos spp. have alsobeen found to be active against amoebae
but in contrast to emetine, they were lesstoxic to cells. Usambarensine,
usambarine, and 18,19
dihydrousambarine from Strychnosusambarensis have been found to be
higWy active against E. histolytica invitro. Nb-Methylusambarensine was
comparatively less activeagainst amoebae
than was usambarensine. Gossypol, a
polyphenol from cotton seed oil, is also
Conclusion
rate of efficacy in terms of clinical 4.
improvement.
Casinovi C.G., Fardella G. andGandolini G. (1981). PharmEducation & Science, 36(2),116-22.
Chopra R.N.,Chopra I. C.,Handa KL.
and KapurL.D.,Indigenous Systemof Medicine, U.N. Dhur and Sons
Pvt. Ltd., Calcutta, 1958, 2nd edn,
522,603.
Evans W.C., Trease and Evans'
Pharmacognosy, W. B. Saunders &
Company, London, 2002, 15th edn,407-413.
Anturlikar S.D., Gopumadhavan S.,
Mitra S.K., Chauhan, B. 1. and
Kulkarni, R. D., Indian Drugs,
1993,30(11),582-85.
11. Ghosal S., Prasad B.N. and Lakshmi
v., 1Ethnopharmacology, 1996,50,167-70.
12. Jain P.K,VermaR., KumarN.andAnil
Kumar,1Res Ayurveda & Siddha,1985,6 (1,3,4),88-91.
13. Jamil S., Alam K, Ahmad S., AkhtarS. and All A., 1Sci Pharm, 2002,3(3),99-101.
14. Kuhnt M., Probstle A., Bauer R. and
Heinrich M.,Planta Medica, 1993,59 (Suppl), A-665.
Galvez]., Zarzuelo A., Crespo M.E.,Lorente M. D., Ocete M. A. and
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10. Ghosh T.K, Sen T.,Das A.,DuttaA. S.
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Ansari A.A.,Rafiquddin M. and Aziz
A.,AClinical Studyof Post-e-Bekh,Madar & Marore phali in ZaheerAmoebai. In Dandia P.C. and S.B.
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References
Natural products will continue to
provide novel compounds which may
ultimately lead to new antiamoebic drugs. 6It is important to evaluate locally used .
traditional systems of medicines to
determine their efficacy and safety inclinical situations. There are a number of
drugs of plant-origin in Unani system of 7.
Medicine which have not yet been tried
clinically.It is suggested that after reievent
pharmacognostical and phytochemical
studies (if not done previously), they may
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animals and human subjects. So, that the
patients may benefit from the traditional
wisdom. It is hoped that further research
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on pure compounds for the treatment ofamoebiasis.
Berberis aristata
Boerhaavia diffusa
Laboratories (Delhi), Dawakhana Ajmal
KhanTibbiyyaCollege(Aligarh),etc.These
drug houses also prepare and market the
antiamoebic Unanidrugs. These marketed
preparations are being used by the Unaniphysicians in India and abroad in theclinical situations. These formulations are
based on one or more of the herbal drugs
(mentioned in the foregoing text and
tables) and include Dawa-e-Siyah 3.Pechish, Habb-e-Pechish, Safoof-eMuqliyasa, Sharbat-e-Habbul-As,lawarish Tabashir, lawarish Amla,
lawarish. Mastagi, lawarishSafarjali Qabiz, Majoon SangdanaMurg. These formulations have a high
Natural Product Radiance Vol 2(1) January-February 2003
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Polyherbal Un ani formulation,
Zaght-al-Dam Qavi for hypertensionAclinical evaluation of the efficacyof the polyherbal Unani formulation, Zaght-al-Dam Qavi in hyperten
sion has been done at lamia Hamdard, Hamdard University,NewDelhi. This Unani formulation consists of Tukhm-e-kahu(Lactuca scariola Linn.), Gul-e-Neelofar (Nymphaea alba Linn. ),Asrol (Rauwolfia serpentina Linn.) andKishneez (Coriandrum sativum Linn.). One dose offormulation containing 109 Kishneez, Tukhm-E-Kahu,lOg,
Gul-e-Neelofar, 5 g and Asrol (root) 19was given orally twice a day for the period of 2 months. The formulation wasfound to be effective in the reduction of clinical symptomps of hypertension in the patients i.e., headache, palpitation,
dizziness, fatigue, nervousness, insomania, etc. The blood pressure was normalized after two months of the treatment
[Siddiqui etal, J Sci Pharmacy, 2002, 3(4),154-157].
Natural Product Radiance Vol 2(1) January-February 2003