Review CRYPTOLEPIS SANGUINOLENTA (LINDL.) SCHLECHTER (PERIPLOCACEAE / ASCLEPIADACEAE): A RESOURCEFUL MEDICINAL PLANT M. J. Adisa1*, T. B. Adebayo2 and M. A. Muhammad1 1Department of Chemistry, IBB University, Lapai, Niger State 2Department of Chemistry, Kwara State University, Malete jadisa@rocketmail.com ABSTRACT A plethora of scholarly articles on the therapeutic efficacy of Cryptolepis sanguinolenta reveals its diuretic, anti-malarial, anti-diabetic, anti-aphrodisiac, anti-microbial, anti-thrombotic, anti-inflammatory and anti-cancer activities. The plant has long been employed by Ghanaian traditional healers in the treatment of various fevers including malaria. Several active compounds and their respective salts have been isolated from various parts of the plant. Besides the major secondary metabolite (cryptolepine), all other minor compounds have remarkable and numerous pharmacological activities. They were proven to have a broad spectrum of antipathogenic activity. C. sanguinolenta has potentials in in-vitro antiplasmodial activity against both chloroquine-sensitive and chloroquine-resistance Plasmodium falciparum and moderately cytotoxic. The rationale behind this write-up is to harmonize the rich but scattered information available on this promising medicinal plant of tropical African origin and to stimulate conservative interest in its therapeutic potentials. Keywords: Cryptolepis sanguinolenta, major secondary metabolite (cryptolepine), therapeutic potentials, cytotoxic, tropical African origin. INTRODUCTION The genus Cryptolepis R. Br. is native to tropical Africa and southeast Asia [1]. Cryptolepis belong to the order Apocyales, Family Periplocaceae or Asclepiadaceae. The genus comprises of about 20 species among which are C. sinensis Lour Merr, C. bucanani Roem. Shult, C. Obtusa N. E. Br.,C. sanguinolenta, C. brazzei, C. deciduas, C. oblongifolia, C. triangularis, C. nigritana etc. The first two species mentioned above grow in some parts of southeast Asia and they are used in the treatment of snake bite and scabies, and treatment of chill and edema respectively. The third mentioned is used in Mozambique to relieve abdominal pain [1]. The specie that shows a promising medicinal potential, C. sanguinolenta (Lindl.) Schltr. grows across the tropical Africa, West Tropical Africa: Mali, Senegal, Guniea, Guinea-Bissau, Sierra Leone, Liberia, Cote D’Ivoire, Ghana, Togo, Benin and Nigeria [2]; West-Central Tropical Africa: Cameroon, Central African Republic, Congo, Democratic Republic of Congo (formerly Zaire) [3]; South Tropical Africa: Angola [4] and East Tropical Africa: Uganda, Tanzania [5]. C. sanguinolenta, also known as Pergularia sanguinolenta [6] is a shrub indigenous to West Africa. In Nigeria C. sanguinolenta has been found in Womirere, Iresi, Osun State [7] and Ojurin Akobo- Olorunda road, Oyo, Oyo state [8], In Ghana, the plant is commonly found in Aburi Hills in the Akwapim areas and Ejura distric of the Ashanti region. The plant is given various names in different parts of the country. It is called Gangamau in Hausa, Kadze in Ewe and Nibima in Twi. In Guinea Bissau, a root decoction of this plant has been used for the treatment of various fevers, including hepatitis, and the leaves have been used for the treatment of malaria or powdered as a cicatrizant of wounds [9]. In Ghana, the roots have been used in the clinical therapy of malaria and of urinary and upper respiratory tract infections for at least two decades [10]. Traditionally, extracts of cryptolepis is being used in Guinea-Bissau, Democratic Republic of Congo and Senegal for the treatment of jaundice and hepatitis [11]; stomach and intestinal disorders [12] and amoebiasis [13] respectively. Reports showed that extracts from the root bark of C. sanguinolenta possessed a promosing antiplasmodial activity against the P. falciparum chloroquine sensitive strain D-6 [14]. The in-vitro screening of cryptolepine and its hydrochloride, 11-hydroxycryptolepine and Neocryptolepine against Plasmodium falciparum D-6 (Chloroquine-sensitive strain), K-1, and W-2 (Chloroquine resistant strains) produced a strong antiplasmodial activity against P. falciparum chloroquine resistant strains. Although, the other isolated compound (quindoline) was found to be less active. Cryptolepine has been found to possess other several interesting pharmacological activities as hypotensive [15], antipyretic [16], anti-inflammatory [17], in- vitro antibacterial [14, 18, 19], and antimalarial effects[20]. **Activity of cryptolepine was not limited to only antiplasmodial effects. The major alkaloid in C. sanguinolenta roots, the indoloquinoline alkaloid (cryptolepine) was isolated in 1931 (Delvaux, 1931), and has since been reported present in plant material from Ghana and Nigeria ( Dwuma-Badu et al. 1978). The NMR assignments of its structure have been reported recently (Ablordeppey, 1990). The extensive investigation of the pharmacological activity of cryptolepine has shown hypotensive (Noamesi, 1980), antipyretic (Raymond-Hamet, 1937), anti-inflammatory (Bamgbose and B.K. Noamesi, 1981), in-vitro antibacterial (12, Boakye-Yiadom and S.M. Heman Ackah , 1979), and antimalarial (Noamesi 1991 ) effects. Isolation of another indoloquinoline alkaloid, quindoline from the roots of C. sanguinolenta (6), was reported in 1978 by Dwuma-Badu et al. while the isolation of a third alkaloid from this species, a unique spiro-nonacyclic alkaloid, (cryptospirolepine) came into lime light in 1993(16, A.N. Tackie et al.), while the report of the isolation and structural elucidation of three other indole alkaloids, hydroxycryptolepine, cryptoheptine and cryptoquindoline, was most recent(Alexandra Paulo). The new alkaloid cryptoheptine {4] has revealed antibacterial activity against Gram negative and Gram-positive bacteria (17).** MORPHO-HISTOLOGY Cryptolepis sanguinolenta is a slender, thin-stemmed scrambling shrub with orange-coloured juice in the cut stem [25]. It is a climber which grows better in the rainforest areas than on the plains [26]. The root which is the main source of its anti-malarial activity is bright yellow when it is cut and the root varies from 0.4-6.6 cm in length and 0.31-1.4 cm wide charaterised with a bitter taste. The seeds are oblong in shape, small and pinkish, embedded in long silky hairs. The dried leaves, stems and roots of cryptolepis have a sweet fragrance. The morphological and histological characters to identify the C. sanguinolenta showed the highest leaf index variations of 6.0–9.8cm in length and 3.0–5.3cm in width while the shortest range of 5.0–7.2cm in length and 2.4–3.5cm in width, the petiole longest lenght 1.0–1.7cm and the shortest between 0.6 – 1.1cm. The longest pod length was between 14.8–22.5cm and shortest 10.5–21.3cm; the maximum width 0.5–9.5cm and the minimum 0.4 – 0.9cm. The longest seed length was reported to be 0.6 – 1.0cm and the shortest was 0.6 – 1.3cm [27]. C. sanguinolenta grows wild along the west coast of Africa. Intra-specific variation in some of the edaphic factors supporting the growth of C. sanguinolenta confirmed that the top soil characteristics including soil pH were significantly different from location to location. Report showed that the textural class of soils on which the plant species grows varies from sandy to clay to loam, with sandy between 57.0% - 71.6%, clay 18.78% - 37.12% and loam 5.88% -10.76% [28]. The plant grows on soils that are acidic and near neutral in nature. Figure 1: Cryptolepis sanguinolenta Cryptolepine The major alkaloid of the roots, cryptolepine [22, 28] is reported to possess a multiplicity of biological effects including antimicrobial, antimuscarinic, vasodilating, noradrenergic, antithrombotic, antiinflammatory, and hypoglycemic activities [29]. Some of these effects have been demonstrated in the crude extracts as well as its fractions [26]. The aqueous extracts of the roots have been earlier reported to exhibit antimicrobial activity against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa and Candida albicans [30, 31] as well as antimalarial properties. Of recent is the report of hot water extract and ethanolic extract which were found to show broad spectrum inhibitory activity against both Gram-positive and Gram-negative organisms as well as C. albicans [32]. However, the fact that the hot water extract had comparable zones of inhibition to those of the ethanolic extract attracted the conclusion that the active constituent, cryptolepine may possibly be thermostable. Figure 2: Structure of 1-Cryptolepine C. sanguinolenta has reasonable antimycobacterial activity and is relatively safe. This therefore gives credence for their use in the treatment of tuberculosis by traditional herbal practitioners. A study had nevertheless, proved that C.sanguinolenta is advantageous over rifampicin by being active on the rifampicin-resistant strain of M.tuberculosis which makes it a potential source of a lead compound that could be developed into a drug to tackle the problems of multi-drug resistance [33]. Fatty Acids Components The amount of oil present in Cryptolepis is very small compared to the alkaloidal extract [34]. The biological activity and the medicinal properties of the roots of the plant is therefore chiefly attributed to the alkaloids. The other minor chemical constituents in the roots of C. sanguinolenta, including the fatty acids, might at least play a synergistic role in the biological activities of the plant. The most abundant fatty acids are from the essential dietary fatty acid linoleic acid. Linoleic acids, linolenic acid and arachidonic acid are unsaturated fatty acids that must be present in the human diet and that are used for the synthesis of prostaglandins in the body [35]. This underscores the health and nutritional benefits of the plant besides its medicinal properties. Cytotoxicity A study revealled that C. sanguinolenta could synergize with hypno-sedatives or other CNS depressants and therefore caution needs to be taken in the concomitant administration of the plant with other CNS depressants[36]. Ansah et al. [37] also agrees that the extract is safe for use at doses less than 500 mg/kg. Furthermore, in vitro studies have reported cytotoxicity at the molecular level although may not be reflected in vivo [38]. Ulcerogenic Potentials A study has reported the effect of Cryptolepis sanguinolenta on food consumption and body weight gain [8]. It was observed that Cryptolepis sanguinolenta causes significant increase in food intake and body weight gain, which suggested that the botanical may stimulate the appetite center, causing increase in the food intake. It was also inferred that Cryptolepis sanguinolenta has anabolic effect, which may prevent tissue loss. The study which revealed magnitude and consequence of basal and histamine-stimulated gastric acid secretions of Cryptolepis sanguinolenta, the gastric secretagogue activity of Cryptolepis sanguinolenta was hitherto attributed to the Cryptolepine, and other minor alkaloids present in the botanical,. Alkaloids have been reported to stimulate gastric acid secretion [39]. The increase in gastric acid secretion seen in Cryptolepis sanguinolenta-treatment is attributed to its hypertrophic and hyperplastic effects on gastric parietal cells. Ovulation Inhibitory in Rabbits It has been stablished clearly that cryptolepis treatment prior to ovulation leads to ovulatory failure and post ovulatory treatment leads to reduced number of implants whilst increasing pre-implantation losses [40]. A study has revealled that Cryptolepis and cryptolepine possess potent anti-inflammatory activity mediated by COX-2 inhibition [17, 41]. Cryptolepine also directly obstructs the activity of prostaglandin E2 [42] which is specifically believed to cause ovum release. The mechanism involved was suggested to be similar to that of diclofenac due to the comparable effects of the two in thier anti-inflammatory action and interference with prostaglandin activity. This study was in futherance of the previous inhibitory effects observed with cryptolepis on reproduction in mice [43]. The results which call for caution in the use of cryptolepis in females of child-bearing age. Future Prospects The root decoction of Cryptolepis, has been traditionally used in Ghana for the treatment of malaria. The anti-malarial effect of the decoction has been studied by the Faculty of Pharmacy, Kwame Nkrumah University of Science and Technology in Kumasi and the Centre for Scientific Research into Plant Medicine. Of recent is the manufacture of the tea bag formulation “PHYTO-LARIA” by a company “Phyto-Riker ”. This was, after extensive studies on the safety and toxicity of the tea bag formulation was carried out by The Noguchi Memorial Institute for Medical Research and The faculty of Pharmacy of KNUST has standardized the tea bag formulation.Clinical trial conducted on the teabag formulation [44], showed that Phyto-Laria® completely eliminated parasitaemia within 3 to 5 days in over 90% of patients, with no observable side effects. Fever was reduced within the first 24 hours. Another indicated that efficacy of Cryptolepis is comparable to that of chloroquine and that there is no need for a separate anti-pyretic medication. PHYTO-LARIA® has been shown to be highly effective in the treatment of acute uncomplicated malaria. Because of the bitter taste of the tea, an encapsulated extract that overcome the bitterness was developed. The Ghanaian Food and Drugs Board has been granted approval- in a project that has received funding from the WHO Africa Regional Office- to enable the Noguchi Institute to conduct dose-finding as well as efficacy studies on the capsule formulation. PHYTOSEARCH A public–private partnership is on course seeking financial assistance to carry out further clinical trials and other relevant studies that will result in a scientifically developed indigenous herbal medicine for the treatment of malaria that will be internationally recognized. Cultivation of the plant on a large scale as a raw material source that will provide employment for Ghanaian farmers is a laudable project. [ref] The dose finding study, funded by WHO Africa, is currently underway at the Noguchi Institute, using the capsule formulation, to determine the optimal dose. Additional studies to determine the best technique to yield the highest quality extract, and development of improved assays for standardizing the extract may be a necessary step. Toxicology studies may be done to confirm the safety of the newly formulated product, as well as bioavailability and pharmacokinetic studies. Cost effectiveness will be a primary outcome measure of the trials, after which the Investigator’s Brochure will be completed and presented to WHO, along with an appeal to WHO to recognize and recommend the product for use in the African continent. Farming networks that will out-grow the present production may soon begin in Ghana to expand the cultivation of the plant. Cryptolepis is a plant indigenous to West Africa and is a climber which grows better in the rainforest areas than on the plains. Growing in different areas may also be compared, to ensure uniformity in quality of the final product. It will be important to develop an alternative formulation for infants and young children, whether it is syrup, a suppository or a sub-lingual spray. It may then be important to conduct a dose finding study and a clinical trial to confirm its efficacy in infants and young children. Finally these products may be launched into the market, with marketing studies and an effective promotional and advertising campaign, including post marketing surveillance. It is assuring that Cryptolepis sanguinolenta used in this formulation can clear chloroquine resistant strains of falciparum parasitaemia. This will make malaria treatment affordable and accessible in Ghana. It also establishes scientific basis for the claim of efficacy of Cryptolepis sanguinolenta against malaria at the prescribed dosage. Diane Winn (2012). Phyto-laria® a promising herbal antimalarial from ghana. The herbal division of phyto-riker pharmaceuticals CONCLUSION The World Health Organization (WHO) acknowledges the important role plant medicine- with proven effectiveness and safety- could play in the formal health system and is encouraging research into plant medicine discovery to treat microbial, cardiovascular and other forms of diseases. The motive towards this review is not for Nigerians to highjack this project from Ghanaians, but to partner with Ghana in the cultivation and the supply of the raw plant material. We are hereby appealling to Nigerian government, the pharmaceutical companies and the researchers to partner with Ghana so as to make the products available to the communities that are most vulnerable to malaria in Nigeria. The positive results yielded by C. sanguinolenta against chloroquine resistant strains of falciparum parasitaemia is a welcome develoment, as WHO [45] reported the emergence of malaria parasites resistants to artemisinin in Asia. 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