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Cytotoxicity and antileishmanial activity of Annona muricata pericarp
Fitoterapia 71 Ž2000. 183]186
Short report
Cytotoxicity and antileishmanial activity of Annona muricata pericarp M.C. Jaramillo a , G.J. Arango a,U , M.C. Gonzalez ´ b, S.M. Robledo c , I.D. Velez c a
Facultad de Quımica Farmaceutica, Uni¨ ersidad de Antioquia, A. A. 1226 Medellın, ´ ´ ´ Colombia b Department of Farmacology, Uni¨ ersidad de Valencia, 46100 Burjasot, Spain c Facultad de Medicina (PECET), Uni¨ ersidad de Antioquia, A. A. 1226 Medellın, ´ Colombia
Received 16 March 1999; accepted in revised form 18 September 1999
Abstract Hexane, ethyl acetate and methanol extracts of Annona muricata pericarp were tested in vitro against Leishmania braziliensis and L. panamensis promastigotes, and against cell line U 937. The ethyl acetate extract was more active than the other extracts and even of Glucantime W used as reference substance. Its fractionation led to the isolation of three acetogenins } annonacin, annonacin A and annomuricin A. Q 2000 Elsevier Science B.V. All rights reserved. Keywords: Annona muricata; Acetogenins; Antileishmanial activity; Cytoxicity
Plant. Annona muricata L. ŽAnnonaceae., dried pericarp, collected in Medellın, ´ Colombia, in August 1996 and identified by A. Cogollo ŽHUA.. A voucher specimen ŽNo. AC 013259. is kept at the Herbarium of University of Antioquia. Uses in traditional medicine. Rural inhabitants use the pericarp to treat chronic ulcers w1]4x. U
Corresponding author. E-mail address: [email protected] ŽG.J. Arango A..
0367-326Xr00r$ - see front matter Q 2000 Elsevier Science B.V. All rights reserved. PII: S 0 3 6 7 - 3 2 6 X Ž 9 9 . 0 0 1 3 8 - 0
184
M.C. Jaramillo F. et al. r Fitoterapia 71 (2000) 183]186
Previously isolated classes of constituents. No report. New-isolated constituents. Annonacin Ž0.0032%., annonacin A Ž0.0021. and annomuricin A Ž0.0021. w5]10x from the EtOAc extract.
Annonacin, annonacin A Annomuricin A
RsH R s OH
Annomuricin A. IR bands ŽCHCl 3 .: 1750 cmy1 ŽC s O. and 3400 ŽOH stretch .; H-NMR Ž250 MHz, CDCl 3 .: d 3.8 Ž1H, m, H-4., 3.4 Ž1H, m, H-10., 3.4 Ž1H, m, H-11., 3.35 Ž1H, m, H-15., 3.8 Ž1H, m, H-16., 1.58 Ž2H, m, H-17., 1.97 Ž2H, m, H-18., 3.8 Ž1H, m, H-19., 3.89 Ž1H, m, H-20., 7.19 Ž1H, d, H-33., 5.0 Ž1H, m, H-34., 1.58 Ž3H, d, H-35.; 13 C-NMR: 174.8 ŽC-1., 69.65 ŽC-4., 74.30 ŽC-10., 74.3 ŽC-11., 74.39 ŽC-15., 82.35 ŽC-16., 28.75 ŽC-17., 28.75 ŽC-18., 82.28 ŽC-19., 71.15 ŽC-20., 152.15 ŽC-33., 78.12 ŽC-34., 19.0 ŽC-35.; FABMS MHq 613; loss of H 2 O at m r z 595, 577, 559, 541, and 523 indicates the existence of five OH groups. The positions of the OH groups are suggested by the EIMS ions at m r z 199, 269, 271, 241 and 141. The stereochemistry of the mono THF ring is threo-trans-erythro.
1
Annonacin. 1 H-NMR: d 3.5 Ž1H, m, H-10., 1.5 Ž2H, m, H-11., 3.4 Ž1H, m, H-15., 1.61 Ž2H, m, H-17., 3.41 Ž1H, m, H-19., 3.85 Ž1H, m, H-20., 1.42 Ž3H, d, H-35.; 13 C-NMR: 174.0 ŽC-1., 131.0 ŽC-2., 69.83 ŽC-4., 71.51 ŽC-10., 38.0 ŽC-11., 74.02 ŽC-15., 82.48 ŽC-16., 28.37 ŽC-17., 28. 37 ŽC-18., 82.48 ŽC-19., 74.02 ŽC-20., 151.81 ŽC-33., 77.94 ŽC-34.. MHq FABMS 597; loss of H 2 O at m r z 309, 291, 379, 361. The stereochemistry is threo-trans-threo. Annonacin A. 1 H-NMR: d 1.2 Ž2H, m, H-11., 3.41 Ž1H, m, H-15., 1.63 Ž2H, m, H-17., 3.82 Ž1H, m, H-20., 7.17 Ž1H, d, H-33., 1.4 Ž3H, d, H-35.; 13 C-NMR: 69.85 ŽC-4., 71.69 ŽC-10., 74.49 ŽC-15., 82.16 ŽC-16., 83.19 ŽC-19., 74.26 ŽC-20., 151.89 ŽC-33., 79.29 ŽC-34.. The stereochemistry is threo-trans-erythro. Tested material. Hexane Ž0.89%., EtOAc Ž0.58. and MeOH Ž12.2. extracts. Used microorganisms. Cell line U-937 was cultured in RPMI 1640 medium supplemented with 10% bovine foetal serum and incubated at 378C and 5% CO 2 atmosphere w11x. Leishmania (¨ iannia) braziliensis ŽMHOMrCOr88rUA-301. and
M.C. Jaramillo F. et al. r Fitoterapia 71 (2000) 183]186
185
Table 1 Antileishmanial activity and cytotoxicity of the extracts from Annona muricata pericarp a Tested material
MeOH extract Hexane extract EtOAc extract Glucantime W a
MEC Žmgrml. L. (¨ ) braziliensis
L. (¨ ) panamensis
Cell line U-937
) 1.0 ) 1.0 0.1 4.1
1.0 1.0 1.0 3.7
) 1.0 1.0 0.1 0.46
MEC, minimum effective concentration.
L. (¨ ) panamensis ŽMHOMrCOr87rUA-140. were isolated from patients from the north-west of Colombia with cutaneous injuries, conserved in liquid nitrogen, and cultured in liquid medium ŽSchneider’s insect medium, supplemented with 10% bovine foetal serum. at 268C. Studied activity. Antileishmanial activity and cytotoxicity w11x. Tubes were seeded with either liquid medium with 10 6 promastigotesrml or medium containing 10 6 cell U-937rml, respectively. The minimum effective concentration ŽMEC. was determined after 48 h of incubation at 378C, using Glucantime W Ž N-methylglucamine antimonate. as a reference substance w3,4x. Results. Reported in Table 1. Conclusions. The crude ethyl acetate extract of Annona muricata pericarp was more active than Glucantime W against both tested strains of Leishmania promastigotes and cell line U-937. These results, showing in particular an interesting antileishmanial activity, confirm the importance of the investigation on the therapeutic potential of plants used by rural communities.
Acknowledgements Colciencias, an organization whose objective is to promote scientific and technological development Colombia, financially supported this work. We wish to thank Dr Diego Cortes of the Faculty of Pharmacy, Universidad de Valencia ŽSpain..
References w1x Correa QJE, Bernal HY. Especies vegetales promisorias del convenio andres ´ bello, vol. I, VI. Bogota: ´ Guadalupe Ltda ŽEd., 1989, 1990. w2x Gupta RMP. 270 plantas medicinales Iberoamericanas. CYTED. Bogota: ´ Presencia Ltda ŽEd., 1995.
186
M.C. Jaramillo F. et al. r Fitoterapia 71 (2000) 183]186
w3x Velez BID. Distribution des especies de Leishmania en Colombie. VII Congres ` International de Parasitologie, International Congress of Parasitology. Paris: Bolletin De la Societe ´ ´ Franc¸aise de Parasitologie. Tome 1990;8ŽSuppl 2.:742. w4x Velez BID. El impacto de la leishmaniosis y la enfermedad de chagas en Iberoamerica. Madrid: ´ Jornadas Iberoamericanas de Ciencias Farmaceuticas, Real Academia de Farmacia, 1996:5]30. w5x Rupprecht JK, Hui Y, McLaughlin J. J Nat Prod 1990;53:237. w6x Wu F, Gu Z, Zeng L, Zhao G, Zhang Y, McLaghlin J. J Nat Prod 1995;58:830. w7x Rieser MJ, Gu Z, Fang X, Zeng L, Wood KV, McLaughlin J. J Nat Prod 1996;59:100. w8x Bories C, Loiseau P, Cortes D et al. Planta Med 1991;57:434. w9x Sahpaz S, Bories Ch, Loiseau PM et al. Planta Med 1994;60:538. w10x Cave ´ A. Acetogenins from Annonaceae. . Phytochemistry of plants used in traditional medicine . Oxford: Clarendon Press, 1995:228]248. w11x Sundstrom C, Nilsson K. Intern Cancer 1976;17:565.
Short report
Cytotoxicity and antileishmanial activity of Annona muricata pericarp M.C. Jaramillo a , G.J. Arango a,U , M.C. Gonzalez ´ b, S.M. Robledo c , I.D. Velez c a
Facultad de Quımica Farmaceutica, Uni¨ ersidad de Antioquia, A. A. 1226 Medellın, ´ ´ ´ Colombia b Department of Farmacology, Uni¨ ersidad de Valencia, 46100 Burjasot, Spain c Facultad de Medicina (PECET), Uni¨ ersidad de Antioquia, A. A. 1226 Medellın, ´ Colombia
Received 16 March 1999; accepted in revised form 18 September 1999
Abstract Hexane, ethyl acetate and methanol extracts of Annona muricata pericarp were tested in vitro against Leishmania braziliensis and L. panamensis promastigotes, and against cell line U 937. The ethyl acetate extract was more active than the other extracts and even of Glucantime W used as reference substance. Its fractionation led to the isolation of three acetogenins } annonacin, annonacin A and annomuricin A. Q 2000 Elsevier Science B.V. All rights reserved. Keywords: Annona muricata; Acetogenins; Antileishmanial activity; Cytoxicity
Plant. Annona muricata L. ŽAnnonaceae., dried pericarp, collected in Medellın, ´ Colombia, in August 1996 and identified by A. Cogollo ŽHUA.. A voucher specimen ŽNo. AC 013259. is kept at the Herbarium of University of Antioquia. Uses in traditional medicine. Rural inhabitants use the pericarp to treat chronic ulcers w1]4x. U
Corresponding author. E-mail address: [email protected] ŽG.J. Arango A..
0367-326Xr00r$ - see front matter Q 2000 Elsevier Science B.V. All rights reserved. PII: S 0 3 6 7 - 3 2 6 X Ž 9 9 . 0 0 1 3 8 - 0
184
M.C. Jaramillo F. et al. r Fitoterapia 71 (2000) 183]186
Previously isolated classes of constituents. No report. New-isolated constituents. Annonacin Ž0.0032%., annonacin A Ž0.0021. and annomuricin A Ž0.0021. w5]10x from the EtOAc extract.
Annonacin, annonacin A Annomuricin A
RsH R s OH
Annomuricin A. IR bands ŽCHCl 3 .: 1750 cmy1 ŽC s O. and 3400 ŽOH stretch .; H-NMR Ž250 MHz, CDCl 3 .: d 3.8 Ž1H, m, H-4., 3.4 Ž1H, m, H-10., 3.4 Ž1H, m, H-11., 3.35 Ž1H, m, H-15., 3.8 Ž1H, m, H-16., 1.58 Ž2H, m, H-17., 1.97 Ž2H, m, H-18., 3.8 Ž1H, m, H-19., 3.89 Ž1H, m, H-20., 7.19 Ž1H, d, H-33., 5.0 Ž1H, m, H-34., 1.58 Ž3H, d, H-35.; 13 C-NMR: 174.8 ŽC-1., 69.65 ŽC-4., 74.30 ŽC-10., 74.3 ŽC-11., 74.39 ŽC-15., 82.35 ŽC-16., 28.75 ŽC-17., 28.75 ŽC-18., 82.28 ŽC-19., 71.15 ŽC-20., 152.15 ŽC-33., 78.12 ŽC-34., 19.0 ŽC-35.; FABMS MHq 613; loss of H 2 O at m r z 595, 577, 559, 541, and 523 indicates the existence of five OH groups. The positions of the OH groups are suggested by the EIMS ions at m r z 199, 269, 271, 241 and 141. The stereochemistry of the mono THF ring is threo-trans-erythro.
1
Annonacin. 1 H-NMR: d 3.5 Ž1H, m, H-10., 1.5 Ž2H, m, H-11., 3.4 Ž1H, m, H-15., 1.61 Ž2H, m, H-17., 3.41 Ž1H, m, H-19., 3.85 Ž1H, m, H-20., 1.42 Ž3H, d, H-35.; 13 C-NMR: 174.0 ŽC-1., 131.0 ŽC-2., 69.83 ŽC-4., 71.51 ŽC-10., 38.0 ŽC-11., 74.02 ŽC-15., 82.48 ŽC-16., 28.37 ŽC-17., 28. 37 ŽC-18., 82.48 ŽC-19., 74.02 ŽC-20., 151.81 ŽC-33., 77.94 ŽC-34.. MHq FABMS 597; loss of H 2 O at m r z 309, 291, 379, 361. The stereochemistry is threo-trans-threo. Annonacin A. 1 H-NMR: d 1.2 Ž2H, m, H-11., 3.41 Ž1H, m, H-15., 1.63 Ž2H, m, H-17., 3.82 Ž1H, m, H-20., 7.17 Ž1H, d, H-33., 1.4 Ž3H, d, H-35.; 13 C-NMR: 69.85 ŽC-4., 71.69 ŽC-10., 74.49 ŽC-15., 82.16 ŽC-16., 83.19 ŽC-19., 74.26 ŽC-20., 151.89 ŽC-33., 79.29 ŽC-34.. The stereochemistry is threo-trans-erythro. Tested material. Hexane Ž0.89%., EtOAc Ž0.58. and MeOH Ž12.2. extracts. Used microorganisms. Cell line U-937 was cultured in RPMI 1640 medium supplemented with 10% bovine foetal serum and incubated at 378C and 5% CO 2 atmosphere w11x. Leishmania (¨ iannia) braziliensis ŽMHOMrCOr88rUA-301. and
M.C. Jaramillo F. et al. r Fitoterapia 71 (2000) 183]186
185
Table 1 Antileishmanial activity and cytotoxicity of the extracts from Annona muricata pericarp a Tested material
MeOH extract Hexane extract EtOAc extract Glucantime W a
MEC Žmgrml. L. (¨ ) braziliensis
L. (¨ ) panamensis
Cell line U-937
) 1.0 ) 1.0 0.1 4.1
1.0 1.0 1.0 3.7
) 1.0 1.0 0.1 0.46
MEC, minimum effective concentration.
L. (¨ ) panamensis ŽMHOMrCOr87rUA-140. were isolated from patients from the north-west of Colombia with cutaneous injuries, conserved in liquid nitrogen, and cultured in liquid medium ŽSchneider’s insect medium, supplemented with 10% bovine foetal serum. at 268C. Studied activity. Antileishmanial activity and cytotoxicity w11x. Tubes were seeded with either liquid medium with 10 6 promastigotesrml or medium containing 10 6 cell U-937rml, respectively. The minimum effective concentration ŽMEC. was determined after 48 h of incubation at 378C, using Glucantime W Ž N-methylglucamine antimonate. as a reference substance w3,4x. Results. Reported in Table 1. Conclusions. The crude ethyl acetate extract of Annona muricata pericarp was more active than Glucantime W against both tested strains of Leishmania promastigotes and cell line U-937. These results, showing in particular an interesting antileishmanial activity, confirm the importance of the investigation on the therapeutic potential of plants used by rural communities.
Acknowledgements Colciencias, an organization whose objective is to promote scientific and technological development Colombia, financially supported this work. We wish to thank Dr Diego Cortes of the Faculty of Pharmacy, Universidad de Valencia ŽSpain..
References w1x Correa QJE, Bernal HY. Especies vegetales promisorias del convenio andres ´ bello, vol. I, VI. Bogota: ´ Guadalupe Ltda ŽEd., 1989, 1990. w2x Gupta RMP. 270 plantas medicinales Iberoamericanas. CYTED. Bogota: ´ Presencia Ltda ŽEd., 1995.
186
M.C. Jaramillo F. et al. r Fitoterapia 71 (2000) 183]186
w3x Velez BID. Distribution des especies de Leishmania en Colombie. VII Congres ` International de Parasitologie, International Congress of Parasitology. Paris: Bolletin De la Societe ´ ´ Franc¸aise de Parasitologie. Tome 1990;8ŽSuppl 2.:742. w4x Velez BID. El impacto de la leishmaniosis y la enfermedad de chagas en Iberoamerica. Madrid: ´ Jornadas Iberoamericanas de Ciencias Farmaceuticas, Real Academia de Farmacia, 1996:5]30. w5x Rupprecht JK, Hui Y, McLaughlin J. J Nat Prod 1990;53:237. w6x Wu F, Gu Z, Zeng L, Zhao G, Zhang Y, McLaghlin J. J Nat Prod 1995;58:830. w7x Rieser MJ, Gu Z, Fang X, Zeng L, Wood KV, McLaughlin J. J Nat Prod 1996;59:100. w8x Bories C, Loiseau P, Cortes D et al. Planta Med 1991;57:434. w9x Sahpaz S, Bories Ch, Loiseau PM et al. Planta Med 1994;60:538. w10x Cave ´ A. Acetogenins from Annonaceae. . Phytochemistry of plants used in traditional medicine . Oxford: Clarendon Press, 1995:228]248. w11x Sundstrom C, Nilsson K. Intern Cancer 1976;17:565.