Karla M. Santos Ocasio Katia Rodriguez Claudia Ospina, PhD Mayra Pagan, PhD
Outline
Croton discolor
General Description:
Family:
Euphorbiacae Distribution: Native of the Antilles Common name: lechecillo Traditional Uses: Use as tea for coughs Oils use as treatment for rheumatism and leukemia Use as pesticide in crops
Croton discolor
HO H3CO NH
HO H3CO NCH3
O crotonosine (1)
Figure 2. Alkaloids Isolated from Croton discolor
HO
discolorine (2)
Study Aims
To expand to the phytomedicinal knowledge of native and endemic plants of Puerto Rico and to their chemotaxonomy. To determine cytotoxic activity of Croton discolor using Brine Shrimp Lethality Test.
General Methods
Chemical Analysis
Biological Test
Plant Collection
Gunica
Croton discolor
February 23, 2008
Figure 3. Plant Collection Map
Extractions
Plant Drying and Maceration with a mixture of CH2Cl2/MeOH (1:1) Crude Extract Suspended in Water and Extracted with solvents of different polarity
Hexane Chloroform
Extractions
Table 1. Extraction Procedure Data of Leaves and Bark Plant Extract Solvent Used 4000 ml 600 ml 600 ml 600 ml Weight
Column Chromatography
Thin
solvent reservoir
layer chromatography (TLC) was performed with different solvents Hexane and ethyl acetate 9:1better solvent detected to separate compounds
angel hair
Biological Test
Table 2. Brine Shrimp Lethality Data of Croton discolor Plant Previous work with C.discolor: Plant Extract
LC 50 value in g/ml
112 132 >200 >200 83 141 174
Cytotoxic ?*
Chemical Analysis
Figure 6. 1H NMR Spectrum (400 MHz) of Hexane Extract (bark) in CDCl3
Alyphatic Alyllic
Chemical Analysis
Figure 7. 1H NMR Spectrum (400 MHz) of Chloroform Extract (bark) in CDCl3
Conclusion
The extracts of the leaves and bark C. discolor, exhibited LC50 values below 200 g/mL. The most promising activity of the leaves was displayed by crude extract, 112 g/mL and hexane extract 132 g/mL. The hexane and crude extracts were active against two breast cancer cells (MCF-7, T47D), showing a percent of growth inhibition > 80. The chloroform and hexane spectra are charaterized by the presence of alyphatic, alyllic and vinyllic protons.
Future Projects
Subsequent isolation and identification of the active constituents is needed. Testing against specific breast cancer cell lines.
References
Ospina, C. A.; Pagn, M.; Carvajal, A.; Claudio, K; Rivera, J.; Ortiz, I.; Hernndez, J. In Cytotoxic Screening of Tropical Plants Using Brine Shrimp Lethality Test.; Montes, E. L.; Eds.; Cuadernos de Investigacin Number 7; Instituto de Investigaciones Interdisciplinarias: Cayey, 2009; 1-20.
Meyer, B. N.; Ferrigni, N. R.; Putnam, J. E.; Jacobsen, L. B.; Nichols, D. E.; McLaughlin J. L. Brine Shrimp: A Convenient General Bioassay for Active Plant Constituents Planta Mdica 1982, 45, 31-34.
Sam, T. W. Toxicity Testing Using the Brine Shrimp: Artemia Salina. Colegate, S. M. and Molyneux, R. J. Eds. Bioactive Natural Products Detection, Isolation, and Structural Determination. CRC Press, Boca Ratn, FL. 1993, 442-456. Newman, D. J.; Crag, G. M. Natural Products as Sources of New Drugs over the Last 25 Years J. Nat. Prod., 2007, 70, 461-477.
Melndez, P. A.; Capriles, V. A. "Molluscicidal Activity of Plants from Puerto Rico" Ann. Trop. Med. Parasitol., 2002, 96, 209-218
Acknowledgements
PR LSAMP Interdisciplinary Investigation Institute of UPR- Cayey RISE Program at UPR-Cayey Dean of Academic Affairs UPR-Cayey Chemistry and Biology Departments and technicians Melvin De Jesus- technician in Department of Chemistry of UPR- Humacao All members of the Ospina-Pagn Research Group Augusto Carvajal , M.S UPR - Cayey