Catharanthus roseus (PROSEA)

Plant Resources of South-East Asia Introduction

List of species

1, flowering twig; 2, flower; 3, base and top of corolla tube in longitudinal section; 4, fruit; 5, seed (Achmad Satiri Nurhaman)

Catharanthus roseus (L.) G. Don

Protologue: Gen. hist. 4(1): 95 (1837).

Family: Apocynaceae

Chromosome number: 2n= 16

• Vinca rosea L. (1759),
• Lochnera rosea (L.) Reichenb. ex Endl. (1838).

• Madagascar periwinkle (En)
• Indonesia: bunga serdadu, kembang tembaga, tapak dara (general)
• Malaysia: kemunting china, rumput jalang, tahi ayam (Peninsular)
• Philippines: chichirica (Sp), kantotai, amnias (Tagalog)
• Thailand: nom in (Surat Thani), phakpot bok (northern), phaengphuai bok (Bangkok)
• Vietnam: cây bông dừa, dừa cạn, hoa hải dằng.

C. roseus belongs to a small genus of 8 species, all originating from Madagascar except for C. pusillus (Murr.) G. Don, which is restricted to India and Sri Lanka. For centuries, Madagascar periwinkle has been cultivated as an ornamental throughout the tropics and occasionally in the subtropics; it has become naturalized in many regions. It was brought into cultivation in the first half of the 18th Century in Paris, from seeds collected in Madagascar, and was later distributed from European botanical gardens to the tropics as an ornamental.

In traditional medicine, a decoction of all parts of the plant is used to treat malaria, diarrhoea, diabetes, cancer and skin diseases. Madagascar periwinkle is well known as an oral hypoglycaemic agent. Extracts prepared from the leaves have been used as an antiseptic agent for the healing of wounds, against haemorrhage and as a mouthwash to treat toothache. Madagascar periwinkle is also considered to be a diaphoretic and diuretic and is used to relieve indigestion, dyspepsia, dysentery, toothache and wasp stings, and as a vomitive, purgative, vermifuge, depurative and haemostatic.

The aerial parts of the plant are used for alkaloid extraction (vincristine, vinblastine, vindesine, vinorelbine). The alkaloids are prescribed in anti-cancer chemotherapy, usually as part of complex chemotherapy protocols. They are administered intravenously, via injection or infusion. Vincristine (sulphate) is indicated in the treatment of acute leukaemia, Hodgkin's disease, non-Hodgkin's lymphoma, small-cell bronchial cancer, neuro and nephroblastomas, metastasized breast cancer and various sarcomas (especially rhabdomyosarcoma). A normal dose for adults is 1-1.4 mg/m² of body surface area, usually at a frequency of once a week, or once monthly in combination chemotherapy. The indications for vinblastine (sulphate) are Hodgkin's disease, non-Hodgkin's lymphoma, advanced testicular cancer, Kaposi's sarcoma, and sometimes choriocarcinomas and some cases of histiocytosis (especially the Lettere-Siwe syndrome). A normal dose for adults is 4-6 mg/m² of body surface area weekly. Vindesine, a semisynthetic derivative of vinblastine, is indicated in the treatment of acute lymphatic leukaemia (especially in children) and refractory lymphomas and melanomas. When used alone, a normal dose of vindesine is 3 mg/m² of body surface area every 7-10 days for one month, and then at intervals of 15 days. Combination chemotherapy protocols often allow lower doses. The semisynthetized vinorelbine has breast cancer and bronchial cancer as current indications, and is often administered to adults in a dose of 25-30 mg/m² of body surface area in monotherapy, and in lower doses in combination chemotherapy protocols. The major toxic side effect of vinblastine is myelosuppression, especially leukopenia. Neurotoxicity is the dose-limiting toxicity of vincristine. Both neurotoxicity and myelosuppression are observed after administration of vindesine, but they are less severe than with vincristine and vinblastine. All four agents may induce thrombocytopaenia and alopecia.

The dried root is an industrial source of ajmalicine (raubasine), which increases the blood flow in the brain and peripheral parts of the body. Preparations of it are used to treat the psychological and behavioural problems of senility, sensory problems (dizziness, tinnitus), cerebrovascular accidents, cranial traumas and their neurological sequelae.

Madagascar periwinkle is a popular garden ornamental, grown as a perennial in tropical regions and as an annual in temperate regions. It is valued for its bushy habit and large flowers carried above dark green foliage.

The dimeric alkaloids extracted from the aerial parts of Madagascar periwinkle are marketed as a lyophilisate or a solution of a salt designed for the sole intravenous route (direct intravenous or through infusion tubing). Vindesine and vinorelbine, which are semisynthetized derivatives of vinblastine, are marketed as a sulphate and a bitartrate, respectively, for injectable solutions. All these drugs are prescription drugs that pharmacists in western countries cannot issue without the direct authorization of a physician.

The price of vincristine was reported to be over US$ 200 000/kg in 1993; the world market consumes 250-300 kg annually with a value of US$ 50-80 million.

Madagascar periwinkle has been found to contain as many as about 100 constituents with an indole or dihydroindole structure. The principal constituent is vindoline (up to 0.5%); other major compounds are serpentine, catharanthine, ajmalicine (raubasine), akuammine, lochnerine and tetrahydroalstonine. Ajmalicine and serpentine are essentially present in the roots, whereas catharanthine and vindoline accumulate in aerial parts. The aerial parts contain 0.2-1% alkaloids.

The substances of pharmacological interest are the dimeric alkaloids which show a coupling of an indole and a dihydroindole. Of the separate parts, the indole/dihydroindole moiety is derived from the amino acid tryptophan, which is coupled to a monoterpene residue. Several of these dimeric alkaloids have cytostatic properties, but they occur in very small amounts: vincristine (= leurocristine) in up to 3 g/t of dried drug, and vinblastine (= vincaleucoblastine) in a slightly larger amount. Other active compounds are leurosidine (= 20'-epivinblastine) and leurosine (= 15',20'-epoxyvinblastine).

Vincristine and vinblastine are antimitotics. They bind to tubulin and prevent the formation of microtubules that assist in the formation of the mitotic spindle; in this way, they block mitosis in the metaphase. These compounds have a non-trivial toxicity; they both have neurotoxic activity (especially vincristine) because the microtubule assembly also plays a role in neurotransmission. Their peripheral neurotoxic effects are neuralgia, myalgia, paresthesia, loss of the tendon reflexes, depression and headache, and their central neurotoxic effects are convulsive episodes and respiratory difficulties. Other side-effects are multiple and include alopecia, gastro-intestinal distress including constipation, buccal ulcerations, amenorrhoea and azoospermia. As vinblastine is highly leucopenic, its dosage must be carefully controlled. The alkaloids are very irritating; if extravasation accidentally occurs there is a risk of tissue necrosis. It is possible to limit the side-effects by carefully guiding the dose and administration, and intensively monitoring the treatment. In common with all teratogenic chemotherapeutics, pregnancy and breast-feeding are strictly contra-indicated. Semisynthetic derivatives whose structure is closely related to that of the naturally occurring dimeric alkaloids are also used as anti-cancer drugs. Vindesine can be prepared from vinblastine, and it is also a potent antimitotic. Its side-effects include a transient granulocytopaenia and effects comparable to those caused by vincristine and vinblastine, although the neurological symptoms are less obvious. Vinorelbine (= noranhydrovinblastine) is obtained from anhydrovinblastine. It acts preferentially on mitotic microtubules and not so much on neuronal microtubules, and consequently its neurological toxicity is limited. However, its haematotoxic actovity is substantial, so its dosage must be carefully controlled.

Some of the alkaloids (e.g. leurosine and vindoline) exhibit a moderate hypoglycaemic action. However, most experiments to confirm the reputed positive effect on diabetes have had disappointing results. Vinblastine markedly inhibited in vitro the reproduction of Trypanosoma cruzi, the organism responsible for Chagas' disease, which is a major health problem in Central and South America.

Roots to be used in pharmacy must contain at least 0.4% ajmalicine and serpentine (the quaternary base corresponding to ajmalicine; ajmalicine can be derived from it). These compounds can easily be characterized and quantified by two thin-layer chromatography analyses. Ajmalicine is an α-adrenergic blocking spasmolytic, which at high doses reverses the effects of adrenaline and moderates the activity of the vasomotor centres, especially in the brain stem. It temporarily increases the blood flow to the brain.

Antiviral activity has been reported in vitro for some Catharanthus alkaloids. Extracts showed fungicidal activity (e.g. against Fusarium solani that causes wilt in aubergine and Sclerotium rolfsii that causes damping-off disease in tomato) and nematicidal activity (e.g. against Meloidogyne incognita and M. javanica).

Ajmalicine and derivatives are also found in other Apocynaceae, such as Rauvolfia spp.

• An erect or decumbent, deciduous undershrub up to 100(-200) cm tall, usually with white latex; roots up to 70 cm long; stems often woody at base.
• Leaves decussate, simple, elliptical to obovate or narrowly obovate, (3-)4-9 cm × (1-)1.5-3.5 cm, herbaceous to thinly leathery, cuneate and sometimes oblique at base, obtuse or acute at apex with a mucronate tip, entire, glossy green above and pale green below, laxly pubescent to glabrous on both sides, secondary veins 7-11 on both sides of midrib and more or less conspicuous, tertiary venation inconspicuous; petiole (0.1-)0.3-1 cm long, with a fringe of colleters in the axil; true stipules absent.
• Inflorescence terminal, but apparently lateral because of alternating development of one of the axillary buds of the apical leaf-pair, 1-2-flowered.
• Flowers actinomorphic, bisexual, 5-merous, subsessile; sepals slightly connate at base, (2-)3-5 mm × 1-1.5 mm, green; corolla salver-shaped, pink, rose-purple or white with a purple, red, pink, pale yellow or white centre, tube 2-3 cm long and widening near the top, laxly puberulous to glabrescent outside, with a densely strigose ring of hairs in the throat and with a sericeous ring of hairs lower down the tube, lobes broadly obovate, 1-2(-3) cm long, mucronate at apex, glabrous, spreading, in bud overlapping to the left; stamens included in the corolla tube, inserted just below the corolla throat, filaments very short, anthers free, introrse; ovary superior, consisting of 2 very narrowly oblong carpels coherent at base, style filiform, with a cylindrical pistil head provided at base with a reflexed hyaline frill ("petticoat") and with rings of woolly hairs at base and apex, stigma glabrous; disk composed of 2 glands, often longer than ovary.
• Fruit composed of 2 cylindrical and acute follicles 1-4 cm long, striate, laxly puberulous to glabrous, green, dehiscent at adaxial side, many-seeded.
• Seeds oblong, 1-2 mm long, with rugose testa and lateral hilum, black; cotyledons flat, slightly shorter than radicle; endosperm scanty.

Madagascar periwinkle is self-incompatible. The seeds usually fall close to the mother plant, but are sometimes transported by ants.

Catharanthus is very closely related to Vinca but differs in general appearance (Vinca produces trailing or floppy, long-lived stems with persistent, leathery leaves) and in flower characteristics (flowers in Vinca solitary, corolla infundibular, stamens with long filaments, glands at base of ovary small, pistil head without "petticoat").

The flower types present in Madagascar periwinkle differ in whether the corolla is pink, rose-purple, white, or white but red-eyed. Usually, no qualitative differences have been found in alkaloid composition between these flower types, but tests in the Philippines showed that plants with rose-purple flowers had a higher alkaloid production than white-flowered plants. Likewise, tests in Thailand revealed the white red-eyed plants to produce the highest yields of alkaloids and vinblastine. Nowadays, these types are usually classified as cultivars, e.g. cv. Albus.

C. lanceus (Bojer ex A.DC.) Pichon is used medicinally in Madagascar and South Africa; the leaves are used as astringent and emetic, the aerial parts as galactagogue and vomitive, and the roots as purgative. The alkaloid fractions of C. lanceus have shown hypotensive activity (mainly caused by yohimbine, a potent α-adrenergic blocker). A lyophilized aqueous extract was found to show anti-tumour activity (leurosine is most potent), whereas some alkaloids have a hypoglycaemic effect.

Madagascar periwinkle often occurs in sandy locations along the coast, but also inland on river banks, in savanna vegetation and in dry waste places and roadsides, sometimes in open forest or scrub, usually on sandy soils, but sometimes also on rocky soils. It is highly salt-tolerant, and is mostly found near sea-level, but occasionally up to 1500 m altitude. It can stand drought well, but not severe heat. Under severe water stress the alkaloid content of mature leaves was found to double, but it did not change in stems and immature leaves and it decreased in roots.

Madagascar periwinkle is usually propagated by seed. Seed may remain dormant for several weeks after maturity. The optimum temperature for germination is about 25 °C. The germination rate of fresh seed is reported as 40%, but pretreatment with chemical stimulants like a potassium nitrate solution may enhance it to 90%. Madagascar periwinkle can also be propagated vegetatively by greenwood or semi-ripe cuttings rooted in a closed container with bottom heat.

Callus tissue of Madagascar periwinkle can be cultured on supplemented White's medium or supplemented Heller's medium, usually solidified with agar. Tests in Thailand showed that the best callus initiation and greatest weight of callus from stem explants was obtained in a Murashige and Skoog basal medium supplemented with 0.5 mg/l 2,4-D and 1 mg/l kinetin. In the Philippines, successful trials have been carried out using floral explants.

Cell suspension cultures are grown in Gamborg's B5 medium or in LS medium containing 2 mg/l naphthalene acetic acid, 0.2 mg/l kinetin and 30 g/l sucrose, under constant light at 25 °C and constant shaking. Cell cultures of Madagascar periwinkle produce a variety of monoterpenoid alkaloids. The alkaloid spectra of root and shoot cultures are similar to those of roots and aerial parts, respectively, of whole plants. Ajmalicine, serpentine and catharanthine are usually the major constituents. Much higher yields of serpentine and ajmalicine, the hypotensive agents, can be produced in cell cultures than in whole plants: up to 2% on dry weight basis versus 0.3% in whole plants. The dimeric anti-cancer alkaloids vinblastine and vincristine are almost undetectable in cultured cells, so attention has turned to the production of catharanthine and vindoline, which can be used as precursors for their synthesis. However, in experiments the production of vindoline appeared not to be stable in cell culture. The lack of vindoline accumulation in cell suspension cultures has been correlated with the lack of expression of the enzymes which catalyse the last steps of vindoline biosynthesis. These enzymes are considered to express only at later development stages and occur in above-ground plant parts; the last steps in biosynthesis only occur in seedlings grown in light.

Multiple shoot cultures induced from seedlings produce vindoline and catharanthine in rather higher levels. Murashige and Skoog medium supplemented with 7 mg/l benzyladenine and 1 mg/l α-naphthalene acetic acid strongly stimulates the formation of shoots, whereas medium supplemented with 2,4-dichlorophenoxyacetic acid suppresses the formation of shoots.

Another possible method of vindoline production is by cultures of selected hairy roots. These can be established by infecting seedlings with Agrobacterium rhizogenes. Some clones not only showed levels of ajmalicine, serpentine and catharanthine comparable to those of cell suspension cultures, but also about 3 times more vindoline than usually found in cell cultures.

In South-East Asia, Madagascar periwinkle is usually cultivated as an ornamental; there are no records of large-scale cultivation for medicinal purposes. It responds well to N fertilizers, but can also grow and persist on poor soils.

In India, Madagascar periwinkle is largely cultivated as a 200-day crop for its leaves (for the extraction of vinblastine and vincristine) and its roots (for the extraction of ajmalicine). The crop needs little irrigation and fertilizer.

In Malaysia, Madagascar periwinkle has been reported to be infected with so-called Malaysian periwinkle yellow. Symptoms include excessive yellowing of foliage, virescence, phyllody, bunchy top and stunted flowers and leaves, suggesting infection by a mycoplasm-like organism. Similar diseases have been reported from China, Taiwan, North America and Europe. Mycoplasm-like organisms can be transferred to Madagascar periwinkle by parasitic plants of the genus Cuscuta, and perhaps also by leafhoppers. Container-grown plants in the United States cultivated as ornamentals have been reported susceptible to Phytophthora parasitica that causes root and stem rot.

The alkaloids in Madagascar periwinkle used in cancer chemotherapy occur in very small amounts: vincristine in an amount of up to 3 g/t of dried drug, and vinblastine in a slightly larger amount. These dimeric alkaloids are almost undetectable in cultured cells. The serpentine and ajmalicine production can be much higher in cell cultures than in whole plants: 1.3% on a dry weight basis versus 0.3% in whole plants.

The aerial parts of Madagascar periwinkle are separated from the roots; both are dried at low temperatures, then packed for shipment. Potted plants for use as ornamentals are usually traded in sealed packages. They are marketable in this condition for 18 days, and do not require watering during this period.

Although Madagascar periwinkle probably originated from a limited area in south-eastern Madagascar, it is now widely planted and naturalized in all tropical areas, and is certainly not endangered. However, protection of the wild populations in Madagascar is desirable to ensure the conservation of the genetic diversity, which might be of interest for breeding purposes in the future.

Tetraploid plants, induced with colchicine, have been found to have a much higher alkaloid content than diploid plants, but the doubling of chromosomes was found to result in reduced pollen fertility and poor seed set.

C. roseus has been successfully crossed with C. trichophyllus (Baker) Pichon, with the F₁ having a high seed set and good viability when C. trichophyllus was the female parent. The alkaloid profiles of the two species are different, and alkaloid production seems to be higher in hybrids than in the parent species. A possible strategy for improving alkaloid production in Catharanthus could be to breed for hybrids with a high alkaloid content.

The possibility of accessing active dimeric alkaloids by biomimetic synthesis has recently attracted much attention. It is now conceivable that vinblastine could be obtained from starting materials such as catharanthine and vindoline that are neither rare nor too expensive. These latter two compounds can be produced in sufficient amounts in in vitro cultures of Madagascar periwinkle. Studies on analogues of the well-known alkaloids suggest good prospects for new developments vis-à-vis Catharanthus alkaloids.

Madagascar periwinkle may have some prospects as a protectant of stored grain, since tests in the Philippines have demonstrated that its use results in improved germination of treated maize kernels and in vigorous seedlings. Corn weevil and flour beetle infestation were also reduced. However, possible toxicity and teratogenicity of residues could be a serious drawback.

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• H. Sutarno & Rudjiman