African Journal of Pharmacy and Pharmacology Vol. 5(20), pp. 2292-2305, 29 November, 2011 Available online at http://www.academicjournals.org/AJPP DOI: 10.5897/AJPP11.593 ISSN 1996-0816 ©2011 Academic Journals

Full Length Research Paper

Enlisting the scientifically unnoticed medicinal plants of Pakistan as a source of novel therapeutic agents showing anti-venom activity

Muhammad Hassham Hassan Bin Asad1, Ghulam Murtaza1, Sami Siraj2, Shujaat Ali Khan1*, Saira Azhar1, Muhammad Sikander Hussain3, Tariq Ismail1, Muhammad Shahzad Hussain4 and Izhar Hussain1

1Department of Pharmacy, COMSATS Institute of Information Technology, Abbottabad 22060, Pakistan. 2Institute of Basic Medical Sciences, Khyber Medical University, Peshawar, Pakistan. 3Department of Chemistry, Bahauddin-Zakariya-University, Multan, Pakistan. 4Department of Physiology, Faculty of Biosciences, University of Veterinary and Animal Sciences, Lahore, Pakistan.

Accepted 23 September, 2011

Snake bite envenoming is a global occupational hazard and most of the people of the world trust in traditional medicine for snake poisoning. The present review elaborates scientifically un-investigated/ ignored medicinal plants of Pakistan showing chemical constituents of natural origin with possible mechanisms showing anti-venom activity. This review enlists 35 plants with their families, distribution in Pakistan, parts used traditionally for snake bite treatment and various active principles present in them. Compositae is the most excessive family, with 3 species, followed by Amaranthaceae, Apocynaceae, Asclepiadaceae, Caesalpinaceae, Labiatae, Pinaceae, Polygonaceae and Verbinaceae having 2 species of medicinal plants. While, one plant species belongs to each, Aizoaceae, Araceae, Boraginaceae, Chenopodiaceae, Cucurbitaceae, Euphorbiaceae, Flacourtiaceae, Gentianaceae, Malvaceae, Menispermaceae, Mimosaceae, Oxalidaceae, Papilionaceae, Plantaginaceae, Salvadoraceae and Solanaceae. As an antidote to snake poisoning, the traditional use of leaves (35%) is higher than roots (25%), whole plant (21%), flower (7%), wood (5%), fruit (5%) and milky juice (2%). Among life forms of plants, herbs (55%) are more excessively used as snake bite remedy than shrubs (31%) and trees (14%). This article may help the researchers to bring novelty in the field of natural products for the treatment of snake bite. However, chemical and pharmacological studies are necessary to confirm the anti-venom claims about these medicinal plants of Pakistan.

Key words: Venom, anti-venom, herbal remedy, scientific characterization.

INTRODUCTION

History of natural products is as old as human civiliza- in order to prevent, mitigate and cure a disease or to alter tions. From ancient time, products from plants were the physiological and pathological processes are known as successful remedies due to their enhanced acceptability medicinal plants. Medicinal plants have an impact in the in human societies, better compatibility with the body and society of developing countries, not only as a source of to treat different ailments due to their synergistic and/or income but also for improving health, because 80% of side effects neutralizing combinations (Rates, 2001). human population rely on herbal remedies (Shah et al., Plants that are source of some drugs or their precussor 2009). Modern pharmacopoeia consists of at least 25% of the drugs that come from plant origin, 121 of such active compounds are being in current use or synthetic analogue are obtained from natural precursors. Hence, *Corresponding author. E-mail: [email protected]. Tel: +92- importance of medicinal plants cannot be under estima- 992-383591-5. Fax: + 92-992383441. ted (Shinwari, 2010).

Asad et al. 2293

Figure 1. Map of Pakistan representing distribution of snakes from genus Naja (A guide to the Onakoo of Pakistan, 2002).

Pakistan has an area 80,943 km2 and it lies between 60° venom induced nephrotoxicity, haemorrhage, local tissue 55‟ to 75° 30‟ E longitude and 23° 45‟ to 36° 50‟ N lati- damage and necrosis. Moreover, antisera development is tude. Moreover, it has an altitude ranges from 0 to 8611 precious, time consuming, tedious and requires strict m, with mixture of climatic zones and distinctive biodi- storage conditions (Razi et al., 2010). Due to these versity of medicinal plants. Pakistan has 6,000 species of drawbacks in antiserum therapy, medicinal use of plants higher plants, 600 to 700 of which are used for medicinal against snake bite has long been recognized because purposes. Majority of Pakistan‟s population depends on traditional healers are first line defense for snake bite folklore remedies for minor and major diseases. Medici- victims. nal plants of Pakistan have potentials to treat any sort of According to the literature survey, various plants have disease from headache to stomachic to cut and wound been reported previously to be active against snake bite. (Shinwari, 2010). The following are the few examples: Acacia catechu Snake bite envenomation is one of the challenging Willd, Belamcanda chinensis DC, Caesalpinia bonduc threats in Pakistan, because 20,000 deaths occur Roxb, Daphne mezereum L, Echinacea angustifolia DC, annually due to this reason and numerous cases of chro- Fagopyrum cymosum Meissn, Gentiana lutea L, nic physical handicaps are reported (Gutiérrez et al., Harpalyce brasiliana Benth, Impatiens balsamina L, 2006). Snakes belonging to genus Naja are Naja naja ( Liatris squarrosa Willd, Macfadyena unguis-cati Gent, ) and Naja oxiana (o) are deadly poisonous and are Nerium oleander L, Ocimum basilicum L, Pentaclethra distributed throughout Pakistan (Figure 1). Cytotoxicity, macroloba (Willd.) Ktze, Ruta graveolens L, Serenoa myotoxicity, oedema, pain, necrosis, haemorrhage, fang repens Small, Taraxacum officinale Weber and marks in the skin, diarrhea, burning, convulsions, dizzi- Verbascum thapsus L (Mors et al., 2000). However, to ness, weakness, blurred vision, fever, excessive the best of our knowledge and according to the literature sweating, tingling and high pulse rate are the major com- search, majority of medicinal plants widespread in plications of snake bite (Razi et al., 2010; Das, 2009). Pakistan have not been scientifically evaluated for their Administration of antisera is the most appropriate anti-venom potentials. Selection of medicinal plants for remedy for snake bite. However, antisera have some side anti-venom activity is very important and crucial step. effects, that is, anaphylactic shock, serum sickness, Selection can be done by several ways including tradi- pyrogenic reactions and it is not useful in protecting tionnal use, chemical content, toxicity, randomized

2294 Afr. J. Pharm. Pharmacol.

selection or amalgamation of different criteria (Rates, plant 21%, flower 7%, Bark (wood) 5%, fruit 5% and milky 2001). In the present article, an effort has been made to juice showed 2% helpfulness in the management of enlist scientifically ignored Pakistani medicinal plants snake poisoning (Figure 3). It was noticed that different having folklore (ethno-botanical) evidences as anti- medicinal plants were found with different life forms venoms. It is very essential, primarily because scientific (habits). Among them, herbs (55%), shrubs (31%) and (chemical and biological) studies of such medicinal plants trees (14%) were conspicuous (Figure 4). It seemed that would be helpful for the discovery of novel therapeutic herbs are more effective in snake envenoming than agents, beneficial in snake poisoning in future, and shrubs and trees. secondly to attract the attention of researchers in order to rationalize their uses as a remedy for snake bite in traditional system of medicine. DISCUSSION

Plants efficacy to neutralize snake venom may be due to DATA COLLECTION certain chemical compounds possessing anti-lethal effects (Mors et al., 2000). Among them steroids, The key words used for the literature search of this article were triterpenes, acids (benzoic acid, cinnamic acid, “Pakistani medicinal plants, anti-venom, ethnobotanical evidences chlorogenic acid and aristolochic acid derivatives), in snake bite and natural products”. Data collection was carried out phenylpropanoid derivatives, curcuminoids, coumarins, through internet search on Science Direct, Google and PubMed flavonoids, polyphenols and polysaccharides have been using biological and chemical abstracts. Selection of plants was isolated and tested as anti-venom previously (Table 2). In focused on their potentials as anti-venom in folklore remedies and their references were studied in details. Search was also made to this article, authors found that folklore reported medicinal enlist already reported anti-venom constituents with possible plants have potentials of anti-venom due to the presence mechanism to support the anti-venom characteristics of medicinal of copious forms of flavonoids, steroids, tannins, triter- plants of Pakistan. The outcome of the results were rechecked and penoides and coumarins, while a single substance does compared with literature. not reproduce the full activity of an extract rather more than one active constituent present in the plants acting synergistically on various target structures (receptors and RESULTS enzymes). Commonly, these chemical constituents are considered „multifunctional‟, in the sense that they exhibit Humans have been using plants as medicines since the more than one pharmacological or biochemical property start of civilization. Thus, the plants have important role in (Table 2). Therefore, multi-func-tionality has built an human life on this planet. The ascendancy of folklore illustration in the capacity of such chemical substances to remedies of medicinal plants as anti-venom has spurred stick to proteins, hence obstruct the functions of many to develop an inventory of such plant species. In this macromolecules. We suggest that these substances are article, 35 medicinal plants occurring in Pakistan and their responsible for observed protection but the time has crucial information have been enumerated in alphabetical come to validate pharmacologically traditional evidences order of plant scientific name, family, place (distribution), of Pakistani medicinal plants as antidote for snake part used, major constituents and references (Table 1). victims. Those species that are distributed in 25 families of which Search was also made to enlist already reported anti- Compositae stood out with maximum representation with venom constituents with possible mechanism to support 3 plants species exhibiting anti-snake venom potentials. the anti-venom characteristics of medicinal plants of Amaranthaceae, Apocynaceae, Asclepiadaceae, Pakistan (Afzal et al., 2009). Caesalpinaceae, Labiatae, Pinaceae, Polygonaceae and Verbinaceae are the families which hold 2 species showing snake envenoming remedy, while, Aizoaceae, Conclusion Araceae, Boraginaceae, Chenopodiaceae, Cucurbitaceae, Euphorbiaceae, Flacourtiaceae, In the present article, folklore use of enlisted 35 medicinal Gentianaceae, Malvaceae, Menispermaceae, plants as anti-snake venom spread in Pakistan are Mimosaceae, Oxalidaceae, Papilionaceae, recorded and documented. Nowadays, importance of Plantaginaceae, Salvadoraceae and Solanaceae were such medicinal plants has been realized because many found with single medicinal plant of such potential (Figure of them proved to be effective as antidote for snake 2). All species are distributed throughout Pakistan in poisoning. However, there is a need for detailed scientific different places like Cholistan, Ther and Interior Sindh verification especially chemical investigations. (Table 1). Literature search revealed that each part of the Furthermore, studies with new active constituents are plant has been used in the treatment of snake biting. essential for the understanding of physiology, pathology Leaves represent 35% anti-snake venom features. and pharmacology of snake bite as well as other related Among the rest parts of the plant, root form 25%, whole disorders.

Asad et al. 2295

Table 1. Scientifically ignored medicinal plants of Pakistan possessing ethno-botanical evidences as antidote in snake poisoning.

Botanical names of S/N Family Occurrence in Pakistan Part used in snake bite Chemical constituents (if any) References plant Leaves and (Ahmad et al., Ageratum inflorescence juice is Precocene-II, precocene-I and β- 1 Compositae District Attock 2006; Kurade et houstonianum Mill used as antidote to caryophyllene. al., 2010) snakebite.

(Baquar, 1989; All parts are Alkaloid, tannins, carbohydrate, Albizia lebbeck (L.) Karachi, Sind, Punjab, Kutch, Chulet et al., 2 Mimosaceae recommended in flavanoids, proteins, echinocystic Benth Indus dalta 2010; Ahmad, snakebite. acid and amino acids. 2007)

(Husain et al., Morgah biodiversity park, Leaves and roots are Fatty oil, butyric acid, phytosterin, 3 Althaea officinalis Linn Malvaceae 2008; Chopra et Rawalpindi used for snake bite. mucilage and starch. al., 1956)

Rutin, 5, 7, 3‟, 4‟, tetrahydroxy (Baquar, 1989; flavonol -3-rhamnoglucoside and Leaves are effective in Ashok et al., 2009; 4 Amaranthus viridus Amaranthaceae Common throughout Pakistan 5, 7, 3‟, 4‟, - tetrahydroxy snake bite. Ahmad and flavonol. Proteins, fat and Javed, 2007) carbohydrates

Alkaloid, flavonoid, saponin, tannins, phenols, hydrocyanic acid, phytic acid, vitamin A, vitamin B , (Shah et al., 2006; Amaranthus hybridus Leaves are used as an 1 5 Amaranthaceae Peshawar vitamin B , vitamin B , vitamin B , Akubugwo et al., L. antidote to snake bite 2 3 6 vitamin C, vitamin E, minerals (Na, 2007) K, Ca, Mg, Fe, Zn, P) and amino acids

(Baquar, 1989; Essential oil, crystalline lactone Jhomi et al., 2005; Artemisia scoparia Punjab plains to 2000 m and Whole plant is used as scoparin, 01-phenyl-penta-2,4- 6 Compositae Chopra, 1956; Walds and Kit Baluchistan antidote to snakebite diyne, beta-pinene, limonene and Ahmad et al., (E)-beta-osimene 2006)

Margala hills National Park Root is used as an (Baquar, 1989; 7 Bauhinia variegate L. Caesalpinaceae Islamabad, sub Himalayan Gum, tannins and fatty oil antidote to snake poison Ahmad, 2009) tracts from Indus eastward

2296 Afr. J. Pharm. Pharmacol.

Table 1. Contd.

(Khan, 2002; Milky latex of plant is Schmid and Calotropin, calotropagenin, 8 Calotropis procera Greater Cholistan desert, Karak applied on snakebite Bencze, 1953; Asclepiadaceae uscharine, calotoxin, calactin and (Wild.) R.Br district area to neutralize Mahran et al., voruscharine poison. 1973; Chopra, 1956)

Mostly found in Lahore- Emodine, oxymethyl- Cassia occidentalis Islamabad Motorway (M-2), Roots are used for (Ahmad, 2007; 9 Caesalpinaceae anthraquinones, toxalbumin, tannic Linn waste places in rawalpindi district snakebite. Baquar, 1989) acid, mucilage and fatty oil. and Baluchistan

Chitral, Swat, Astor, Hazara (Hussain et al., Cedrus deodara G. Wood (bark) is used for Gum, essential oil, cholesterin and 10 Pinaceae division, Murree hills and in 2006, Baquar, Don snakebite. ascorbic acid in needle. Kashmir. 1989)

Chenopodium 11 Chenopodiaceae album Linn Margala hills National Park, Fruit and roots are used Essential oil, carotene and vitamin (Shinwari, 2000; Islamabad. as antidote to snake bite. C. Baquar, 1989)

Cissampelos Sind, Punjab plains up to 2000 Leaves are used for Sepeerine, bebeerine, (Shinwari, 2000; 12 Menispermaceae paperia Linn m. snake bite. cissampelline, hyatin and hyatinin. Baquar, 1989)

(Farrukh et al., Coccinia grandis Lasbela, Karachi and Indus Plant is effective against Triterpenoides, carotenoides, 2008; Baquar, 13 Cucurbitaceae Linn delta. snake biting. flavonoids, alkaloids and fatty acids. 1989; Singh et al., 2007)

Flavonoids with glycosides (Baquar, 1989; (, genkwanin, isovitexin, Enicostemma Fresh plant is grinded Ghosal and Karachi, Sind, Indus delta, Kutch swertisin, saponarin, 5-O- 14 hyssopifolium Gentianaceae and applied on area of Jaiswal, 2006; and Jhellum district. glucosylswertisin, 5-O- (Willd.) Verdoorn snakebite. Chopra, 1956; glucosylisoswertisin), alkaloid Khan, 2009) (erythrocentaurin) and ophelic acid.

Triterpenes are found in latex (Baquar, 1989; Euphorbia Karachi, Sind, Baluchistan and Milky juice of leaves is 15 Euphorbiaceae (euphol, tirucallol, cycloartenol and Afza et al., 1989; caducifolia Haines Indus delta. effective in snakebite. cyclocaducinol). Khan, 2009)

Asad et al. 2297

Table 1. Contd.

Phytosterol, β-sitosterol-β- D-glucopyranoside, Sub-Himalayan zone 600 - Leaves are crushed and (Baquar, 1989; Flacourtia indica ramontoside, butyrolactone, 16 Flacourtiaceae 1000 m, Rawalpindi district, applied topically on the Gnanaprakash et al., (Burm.) Merr. lignan disaccharide, Poonch, Punjab and Swat point of snakebite 2010; Khan, 2002) flacourtin and coumarins (scoparone and aesculetin)

Margala hills National Park Islamabad, Trans-Indus, Leucas capitata Desf. Labiatae Whole plant is used for (Shinwari, 2000; 17 Swat, Rawalpindi, Poonch, Essential oil and alkaloids snake bite Baquar, 1989) Dir, Chitral and Lower Hazara

Ziarat juniper forest Marrubium vulgare Baluchistan, Khuram, Swat, Roots are remedy for the Bitter substances, essential (Flora of Zairat, 2008; 18 Labiatae Linn Thal, Chitral, Fort bite of rattlesnakes oil and marrubin Baquar, 1989) Sandeman

Leaves, roots and Cardiotoxins, neriodorin, (Naqvi et al., 1994; 19 Nerium indicum Mill Apocynaceae Throughout Pakistan flowers are used against nerioderin, odorin and Baquar, 1989) snake bite karabin

(Mizokami et al., 2008; In shady places in Sind, Acid potassium oxalate, Leaves are used in Chopra, 1956; Baquar, 20 Oxalis corniculata Linn Oxalidaceae NWFP, Chitral, Hunza and isoorientin, isovitexin and snakebite 1989; Qureshi et al., Hazara swertisin 2009)

Pinus roxburghii Murree hills, Swat, Dir, Wood and oleoresin is A-carene, B-carene, A- 21 Pinaceae (Baquar, 1989) Sargent Hazara (600 - 1500 m) used in snakebite pinene and B-pinene

Throughout Pakistan, Plantago lanceolata except Sind, Western Roots are remedy for the Plant contains glucoside (Baquar, 1989; Flora 22 Plantaginaceae Linn Himalaya, Salt Range, bite of rattlesnakes aucubin of Zairat., 2008) Waziristan

2298 Afr. J. Pharm. Pharmacol.

Table 1. Contd.

Flavonoids (rhazianosides A and (Baquar, 1989, rhazianosides B), glycosides Treimer and Zenk, (isorhamnetin-3-7-rhamnoside, 1979; Kaneko and Plains between Indus and 3-7-rhamnoside and roblnin), Namba, 1967; Jhellum, Karachi, Sind, Fruit and leaves are triterpenes (Beto-sitosterol, Mg 23 Rhazya stricta Dcne. Apocynaceae Andersen et al., Lasbella,Khyber, Malakand, antidote to snakebite quinate and urosoic acid), 1987; Kaneko and Dargai, Salt Range and Attock. enzymes (strictosidine synthase Namba, 1967; and NADPH dependent Panhwar and tetrahydroaistonine) and Abro, 2007) alkaloids

Rumex hastatus Juice of leaves is used 24 Polygonaceae Hazara division. - (Jan et al., 2008) D.Don for snakebite

Sind, Baluchistan Coasts, South Minerals (Ca, Cu, Fe, Mg, K, Na, (Baquar, 1989; Waziristan, Peshawar, Salt Rumex vesicarius Leaves are used for Zn), proteins, lipids, ascorbic Chopra et al., 25 Polygonaceae Range, Abbottabad, Hazara, Linn snakebite acid, tocopherol, citric acid, malic 1956; Alfawaz, Hasan Abdal, Margalla, up to acid and oxalic acid 2006) 1200 m

Carbohydrates, alkaloid (Almas, 2001; It is found abundantly all over the Leaves and flowers are (salvadorine), steroids, Kamil et.al., 1999, 26 Salvadora persica Salvadoraceae Pakistan used for snake biting terpenoids, saponins, flavonoids a, b, c; et al., (quercetin and ) 2000a, b)

Roots are powdered and (Khan, 2002; Sarcostemma Beta-amyrin, friedelin and 27 Asclepiadaceae Greater Cholistan desert poultice is applied on the Torrance and viminale Linn. viminalol area of snakebite Marais, 2006)

(Nasir , 1978; Baquar, 1989; Smith and Lectins, Dimethyl sulphides, p- Meeuse, 1966; Sauromatum Swat, Hazara, Chikar, Stimulating poultice in caryophyiiene, indole, skatole, 28 Araceae Borg- Karlson et guttatum Schott. Rawalpindi, Murree and Kashmir snake bite ammonia, trimethylamine and al., 1994; Chen primary amines and Meeuse, 1971; Shinwari and Khan, 2000)

Asad et al. 2299

Table 1. Contd.

Solanum Common in waste places from Solanocarpine, xanthocarpum Schard Root is used for (Baquar, 1989; 29 Solanaceae plains to 1500 m; throughout solanocarpidine, solanines and & Wendl snakebite Shah et al., 2006) Pakistan carpesteral.

Taraxacin, taraxacerin, tataxerine, taraxerol, taraxasterol, homotaraxasterol, (Baquar, 1989; Common throughout Punjab Taraxacum officinale Leaves and roots are 3,4 dioxycinnamic acid, Stuart, 1979; 30 Compositae and Baluchistan from 350 - Weber antidote to snakebite flavoxanthin, inulin, citric acid, Chopra, 1956; 6000 m pheyloxyacetic acid, riboflavin, Ahmad et al., 2006) sitosterol, sitosterin, stigmasterol and coumestro.

Trichodesma indicum Plains to 1400 m throughout Leaves are antidote to Fatty acids and non-steroidal (Baquar, 1989; 31 Boraginaceae (Linn) R.Br Pakistan snakebite compounds. Panhwar and Abro, 2007; Hassan et al., 1982)

Mostly distributed in Potwar Plateau, moist mountains and arid/semi-arid mountains Verbena officinalis It is recommended for Glucoside, verbenalin and (Khan et al., 2006; 32 Verbinaceae including Northern Linn snake bite verbenin. Baquar, 1989) Balochistan, Sulemania range, Waziristan, Kurram agency, Gilgit and Chitral

Kallar Kahar, Rawalpindi, Whole plant is used for Hydrocynic acid, vicianine and (Ahmad, 2008; 33 Vicia sativa L. Papilionaceae Hasan Abdal, Abbottabad snake poison glucosides Baquar, 1989)

(Baquar, 1989; Charkotli hills, Batkhela Leaves are used for Alkaloid, essential oil and 34 Vitex negundo Linn Verbenaceae Barkatullah and district, Malakand snake bite nishindine. Hussain, 2009)

Zaleya pentandara (L.) Mostly in the coastal areas of It is recommended for 35 Aizoaceae - (Khan et al., 2006) Jeffery Pakistan snake bite

2300 Afr. J. Pharm. Pharmacol.

Figure 2. Various families with their number of plants having anti-venom potentials.

Figure 3. Percentage of different parts of plants used in the treatment of snake poisoning.

Asad et al. 2301

Figure 4. Different life forms of medicinal plants effective in snake bite.

Table 2. Reported chemical substances possessing anti-venom activity found in various plants.

S/N Class Chemical constituent Possible mechanism / anti lethal effect References Anti-inflammatory Sitosterol (70% protection) Anti-inflammatory Sitosterolin Anti-inflammatory Stigmasterol (analogue of sitosterol) By complex formation Dehydrocholic acid (80% protection) Cholesterol combines with lysolecithin thus 1 Steroids Cholesterol (60% protection) produces antihemolytic effect. (Mors et al., 2000) Corticosteroids Protective mechanism comparable to antivenom Tigogenin (40% protection) steroids due to chemical analogy. Hecogenin (20% protection) Anti-inflammatory α-spinasterol Anti-inflammatory Anti-inflammatory

2302 Afr. J. Pharm. Pharmacol.

Table 2. Contd.

Oleanolic acid, Lupeol (20% protection) Ursolic acid, Taraxerol, Taraxasterol, α-Amyrin, β-Amyrin, Friedelin (40% protection), All possess anti-snake venom and anti- Epifriedelinol, Alnusenone, Betulinic acid (40% protection) inflammatory properties due to five-ring structure 2 Triterpenes Betulinol (40% protection), of a triterpenes and related conformation are (Mors et al., 2000) Bredemeyeroside (80% protection), important for achievement of liable Echinocystic acid, Cycloartenol, Quinovic acid, pharmacophore. Presenegenin, Alnusenone, Gymnemagenin Gypsogenin,

Primveroside 2-hydroxy-4-methoxybenzaldehyde Anti-inflammatory activity 2,4-dihydroxybenzoic acid (83% protection) Tyrosinase inhibitor Protocatechuic acid (80% protection) Rest all of all produces effects 4-O-methyl ether of 2,4-dihydroxybenzoic acid (1) By attaching proteins, occupying critical binding sites. Hydroxy benzoic 4-O-methyl ester of 2,4-dihydroxybenzoic acid (Mors et al., 2000; 3 (2) By conjugation with compounds of primary acid derivatives 4-hydroxy-3-methoxy-benzaldehyde Mason, 1955) metabolites. Veratrum aldehyde (3) By chelation (seizing of metal atoms). Gentisic acid (2,5-dihydroxy- benzoic acid) (4) By phenolases oxidation of phenols to 3,4-dihydroxybenzaldehyde quinones, which ultimately condense with proteins Monomethyl ether of 2,6-dihydroxybenzoic acid (40% resulting in copolymerization. protection)

Caffeic acid and caffeic acid derivatives (verbascosides, Caffeic acid is a strong lipoxygenase inhibitor and Cinnamic acid rosmarinic acid, Myricerone caffeate and oligomers of caffeic antihepatoxic activity. 4 (Mors et al., 2000) derivatives acid). Myricerone caffeate is selective endothelin Ferulic and isoferulic acid receptors antagonist.

3-0-caffeyl-D-quinic acid (caffeic acid esterified with quinic 5 Chlorogenic acid Lipoxygenase inhibitor and anti-hepatoxic activity. (Mors et al., 2000) acid) 60% protection.

Phenylpropanoid 6 Cynarin (1,5-dicaffeoylquinic acid) 20% protection Choleretic and anti-hepatoxic activity. (Mors et al., 2000) derivatives

Asad et al. 2303

Table 2. Contd.

Curcumin (Kiso et al., 1983; Demethoxycurcumin Anti-inflammatory, anti-hepatoxin, anti-mutagenic, Amman and Wahl, 7 Curcuminoids anti-carcinogenic, lipoxygenase and 1990; Ferreira et Bis demethoxycurcumin prostaglandin-endoperoxide synthase inhibition. al.,1992; Mors et al., Ar-turmerone 2000)

Coumarin, Herniarin (7-methoxy-coumarin), Ayapin (6,7- methylenedioxy-coumarin), 7,8-dihydroxycoumarin (7-O-gluco-side of daphnetin), Not clearly known however, inhibitions of various (Brown, 1977; Mors et 8 Coumarins Suberenone, Marmin, enzymes are reported. al., 2000) Bergapten (20% protection) Monoterpenoid dorstenin Fernolin

Primetin (5,8-dihydoxyflavone), Free quercetin (40 - 80% protection), Rutin (quercetin 3-0-rutoside, 20 - 80% protection), Hesperetin, Pinostrobin, , Apigenin (40% Anti-venom activity is due to inhibition of protection), phospholipases A2 and some flavonoids by lipoxygenase inhibition. (Lindahl and Tagesson, 9 Flavonoids Kaempferol, luteolin, diosmetin, isorhamnetin, morin, myricetin, quercitrin and isoquercitrin, isoflavonoids Other reported activities are anti-inflammatory, 1997; Mors et al., 2000) (, 7-O-glucoside of tectorigenin, iridin, 7-O- anti-hepatoxin, anti-hypertensive, anti-arrhytmic, glucoside of irigenin, derricidin produces 70% protection) hypocholesterolemic and anti-allergic in general. Pterocarpans (cabenegrins A-I 22, A-II and edunol), (wedelolactone, produces 40% protection, demethylwedelolactone).

It forms a 1:1 complex with phospholipase A and 2 (Vishwanath et al., 10 Aristolochic acids Aristolochic acid causes a change in the secondary structures of 1987) the proteins.

Persimmon tannin, Vegetable tannins It is due to protein-tannin interaction (enzyme (Haslam, 1989; Mors et 11 Ellagitannins, (poly phenols) inhibiting activity of tannins) al., 2000) Eufhorbins A and B

(Barbi, 1992; Varlen et Mixture of polysaccharides, especially heteroglycan and Antiinflammatory and immunomodulating property 12 Polysaccharides al., 1989; Mors et al., others produce more than 30% protection. can be extended to anti-venom activity. 2000)

2304 Afr. J. Pharm. Pharmacol.

REFERENCES Cambridge University Press, Cambridge, 230; f35; ISBN 0-521- 32189-1 Afza N, Khan AQ, Malik A, Badar Y (1989). Cyclocaducinol, a Hassan M, Ahmad S, Mahmood K (1982): Chemical investigation of cycloartane type triterpene from Euphorbia caducifolia. Trichodesma indicum leaves –I. Non steroidal constituents of Phytochemistry, 28(7): 1982-1984. petroleum ether extract. J. Chem. Soc. Pak., 4: 281-283. Afzal S, Afzal N, Awan MR, Khan TS, Gilani A, Khanum AG, Tariq S Husain SZ, Malik RN, Javaid M, Bibi S (2008). Ethnobotanical (2009). Ethno-botanical studies from Northern Pakistan. J. Ayub. properties and uses of medicinal plants of Morgah biodiversity park, Med. Coll. Abbottabad, 21(1): 52-57. Rawalpindi. Pak. J. Bot., 40(5): 1897-1911. Ahmad M, Khan MA, Zafar M, Sultana S (2006). Ethnomedicinal Hussain M, Shah GM, Khan MA (2006). Traditional medicinal and demography and ecological diversification of some important weeds economic uses of Gymnosperms of Kaghan valley, Pakistan. from district Attack- Pakistan. Pak. J. Weed Sci. Res., 12(1-2): 37-46. Ethnobotanical Leaflets, 10: 72-81. Ahmad SS (2007). Medicinal wild plants from Lahore-Islamabad Jan S, Khan MA, Siraj-ud-din, Murad W, Hussain M, Ghani A (2008). Motorway (M-2). Pak. J. Bot., 39(2): 355-375. Herbal rmidies used for gastrointestinal disorders in Kaghan valley, Ahmad SS, Javed S (2007). Exploring the economic value of NWFP, Pakistan, Pak. J. Weed Sci. Res., 14(3-4): 169-200. underutilized plant species in Ayubia Nnational Park. Pak. J. Bot., Kamil M, Jayaraj AF, Ahmad F, Gunasekhar C, Samuel S, Habibullah 39(5): 1435-1442. M, Chan K (1999). A Rare Natural Product from Salvadora Persica Akubugwo IE, Obasi NA, Chinyere GC, Ugbogu AE (2007). Nutritional Leaves. 2000 Years of Natural Products Research Past, Present and and chemical value of Amaranthus hybridus leaves from Afikpo, Future. Amsterdam, p. 562. Nigeria. Afr. J. Biotechnol., 6(24): 2833-2839. Kamil M, Jayaraj AF, Ahmad F, Gunasekhar C, Samuel S, Habibulla M, Alfawaz MA (2006). Chemical composition of hummayad (Rumex Chan K (2000). Isolation and Identification of a Flavonol Glycoside vesicarius) grown in Saudi Arabia. J. Food Compose Anal., 19: 552- using high speed counters current chromatographic technique from 555. the leaves of Salvadora persica. Pakistan. J. Sci. Ind. Res., 43(4): Almas K (2001). The antimicrobial effects of seven different types of 225-227. Asian chewing sticks. Odonto-Stomatologie Tropicale, 24: 17-20. Kamil M, Jayaraj AF, Ahmad F, Gunasekhar C, Samuel S, Habibullah M Amman HPT, Wahl MA (1990): Pharmacology of Curcuma longa. Kamil M, Jayaraj AF, Ahmed F, Gunasekhar C, Samuel S, Habibullah Planta. Med., 57: 1-7. M, Chan K (2000). Use of High-speed counter current Andersen WK, Omar AA, Christensen SB (1987). Isorhamnetin 3-(2,6 chromatography for separation of flavonol glycoside from the leaves dirhamnopyranosylgalactoside)-7-rhamnoside and 3-(6- of Salvadora persica. The Mona symposium 2000- Natural Products rhamnopyranosylgalatoside)-7-rhamnoside from Rhazya stricta. and Medicinal Chemistry, University of West Indies, Mona Kingston, Phytochemistry, 26: 291-294. Jamaica, pp. 6-11 Ashok KBS, Lakshman K, Jayaveera KN, Nandeesh R, Mani Tripathi Kamil M, Jayaraj AF, Ahmed F, Gunasekhar C, Samuel S, Habibullah SN, Vamshi Krishna N, Manjunath M, Suresh MV (2009). Estimation M, Chan K (1999): Pharmacognostic and Phytochemical studies on of Rutin and Quercetin in Amaranthus viridis L by High Performance Salvadora persica L. J. Pharm. Pharmacol., 51: 227. Layer Chromatograph (HPLC). Ethnobotanical Leaflets, 13: 437-442. Kaneko H, Namba K (1967). Studies on non-alkaloidal constituents of Baquar SR (1989). Medicinal and poisonous plants of Pakistan. 1st Ed. Indian medicinal plant, Rhazya stricta Decaisne. Yakugaku Zasshi, Printas Karachi Pakistan, pp. 118-121. 87(4): 430-433. Barbi NS (1992). ContribuicË aÄo aÁ pesquisa de plantas antiofõÂ Khan FM (2009). Ethno-veterinary medicinal usage of flora of greater dicas: estudo de polissacarõÂ deos presentes na casca de Casearia Cholistan desert Pakistan. Pak. Vet. J., 29(2): 75-80. sylvestris Sw. M.S. thesis, Federal University of Rio de Janeiro. pp. Khan MA, Barth H, Kust GC, Boer B (2006). Sabkha Ecosystems: 32-34. Volume II: The South and Central Asian Countries. Chapter 11: Barkatullah IM, Hussain F (2009). Ethnobotanical studies of plants of Halophytes of Pakistan: Characteristics, distribution and potential Charkotli hills, Batkhela district, Malakand, Pakistan. Front Biol. economic usages. Springer, Netherlands, pp. 129-153. China, 4(4): 539-548. Khan MS (2002). A guide to the Snakes of Pakistan, 11th Ed, Chimaira Borg-Karlson AK, Englund FO, Unelius CR (1994). Dimethyl Publishers, Frankfurt Germany. pp. 76-77. oligosulphides, major volatiles released from Sauromatum guttatum Kiso Y, Suzuki Y, Watanabe N, Oshima Y, Hikino H (1983). and Phallus impudicus. Phytochemistry, 35: 321-323. Antihepatotoxic principles of Curcuma longa rhizomes. Plata. Med., Chen J, Meeuse BJD (1971). Production of free indole by some aroids. 49: 185-187. Acta. Bot. Neerl., 20: 627-635. Kurade NP, Jaitak V, Kaul VK, Sharma OP (2010). Chemical Chopra RN, Nayar SL, Chopra IC (1956). Glossary of Indian Medicinal composition and antibacterial activity of essential oils of Lantana Plants, CSIR Publications, New Delhi, India, pp. 379-381. camara, Ageratum houstonianum and Eupatorium adenophorum. Chulet R, Joseph L, George M, Pradhan P (2010). Pharmacognostic Pharm. Biol., 48(5): 539-544. standardization and phytochemical screening of Albizzia lebbeck. J. Lindahl M, Tagesson C (1997). Flavonoids as phospholipase A2 Chem. Pharm. Res., 2(1): 432-443. inhibitors: importance of their structure for selective inhibition of group Das K (2009). Medicinal plants for snake bite treatment- future focus. II phospholipase A2. Inflammation, 21: 347-356. Ethnobotanical Leaflets, 13: 508-521. Mahran GH, Abdul-Wahab SM, Ahmad MS (1973). Bull Fac Pharm. Farrukh U, Shareef H, Mahmud S, Ali SA, Rizwani GH (2008). Cairo Univ., 12: 151. Antibacterial activities of Coccinia grandis L. Pak. J. Bot., 40(3): Mason HS (1955): Comparative biochemistry of the phenolase complex. 1259-1262. Adv. Enzymol., 16: 105-184. Ferreira LAF, Henriques OB, Andreoni AAS, Vital GRF, Campos MMC, Mizokami H, Tomita-Yokotani K, Yoshitama K (2008). Flavonoids in the Habermehl GG, de Moraes VLG (1992). Antivenom and biological leaves of Oxalis corniculata and sequestration of the flavonoids in the effects of ar-turmerone isolated from Curcuma longa (Zingiberaceae). wing scales of the pale grass blue butterfly, Pseudozizeeria maha. J. Toxicon, 30: 1211-1218. Plant Res., 121: 133-136. Flora of Ziarat: Ethnobotanic and Medicinal Importance, Hertia Mors WB, Nascimento MC, Pereira BMR, Pereira NA (2000). Plant intermedia (Boiss) O.Ktze (2008). UNDP, Pakistan, 30: 37. natural products active against snake bite-the molecular approach. Ghosal S, Jaiswal DK (2006). Chemical constituents of gentianaceae Phytochemistry, 55(6): 627-642. XXVIII: Flavonoids of Enicostemma hyssopifolium (Willd.) Verd. J. Naqvi BS, Shaikh MR, Maleka FA, Shaikh D (1994). Studies of Pharm. Sci., 69(1): 53-56. antibacterial activity of ethanolic extracts from Nericum indicum ans Gnanaprakash K, Madhusudhana CC, Ramkanth S, Alagusundaram Habiscus rosasinensis. J. Islamic Acad. Sci., 7(3): 167-168. M,Gutiérrez JM, Theakston RDG, Warrell DA (2006). Confronting the Nasir, YJ (1978). Araceae. 1-17. In: E. Nasir & S.I. Ali (eds.), Flora of neglected problem of snake bite envenoming: The need for a global West Pakistan. No. 120. National Herbarium, Islamabad. partnership. PLoS Med., 3: 727-731. Panhwar AQ, Abro H (2007). Ethnobotanical studies of Mahal Kohistan. Haslam E (1989). Plant Polyphenols - vegetabele Tannins Revisited (Khirthar National Park). Pak. J. Bot., 39(7): 2301-2315.

Asad et al. 2305

Qureshi R, Waheed A, Arshad M, Umbreen T (2009). Medico- Smith BN, Meeuse BJD (1966). Production of volatile amines and ethnobotanical inventory of tehsil Chakwal, Pakistan. Pak. J. Bot., skatole at anthesis in some Arum lily species. Plant Physiol., 41: 343- 41(2): 529-538. 347. Rates SMK (2001). Plants as source of drugs. Toxicon, 39: 603-613. Stuart M (1979). The Encyclopedia of herbs and herbalism, Grosset and Razi MT, Asad MHHB, Khan T, Chaudhary MZ, Ansari MT, Arshad MA, Dunlap, New York, NY, p. 304. Najam-us-Saqib Q (2010): Antihaemorrhagic potentials of Fagonia Torrance JD, Maráis JLC (2006). Triterpene constituents of cretica against Naja naja karachiensis (black Pakistan Cobra) venom. Sarcostemma viminale R. Br. J. Pharm. Sci., 52(5): 439-441. Nat. Pro. Res. doi: 10.1080/14786419.2010.490785. Treimer JF, Zenk MH (1979). Purification and properties of strictosidine Schmid H, Bencze W (1953). Helv Chim. Acta., pp. 36: 205 synthase, the key enzyme in indole alkaloid formation. Eur. J. Shah GM, Khan MA, Ahmad M, Zafar M, Khan AA (2009). Observations Biochem., 101(1): 225-233. on antifertility and abortifacient herbal drugs. Afr. J. Biotechnol., 8(9): Treimer JF, Zenk MH (1979). Purification and properties of strictosidine 1959-1964. synthase. FEBS Lett., 97(1): 159-162. Shah SRU, Hassan G, Rehman A, Ahmed I (2006). Ethnobotanical Varlen J, Liptak A, Wagner H (1989). Structural analysis of a studies of the flora of district Musakhel and Barkhan in Balochistan, rhamnoarabinogalactan and arabinogalactans with Pakistan. Pak. J. Weed Sci. Res., 12(3): 199-211. immunostimmulating activity from Calendula officinalis. Shah SRU, Qasim M, Khan IA, Shah SAU (2006): Study of medicinal Phytochemistry, 28: 2379-2383. plants among weeds of Wheat and Maize in Peshawar region. Pak. J. Vishwanath BS, Rao AGA, Gowda TV (1987). Interaction of Weed Sci. Res., 12(3): 191-197. phospholipase A2 from Vipera russelli venom with aristolochic acid: a Shinwari MI, Khan MA (2000). Folk use of medicinal herbs of Margalla circular dichroism study. Toxicon, 25(9): 939-946. Hills National Park, Islamabad. J. Ethnopharmacol., 69(1): 45-56. Shinwari ZK (2010): Medicinal plants research in Pakistan. J. Med. Plants Res., 4(3): 161-176. Singh G, Gupta P, Rawat P, Puri A, Bhatia G, Maurya R (2007). Antidyslipidemic activity of polyprenol from Coccinia grandis in high- fat diet-fed hamster model. Phytomedicine, 14(12): 792-798.