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Antimicrobial and cytotoxic constituents from the seeds of Annona squamosa
Fitoterapia 76 (2005) 484 – 489 www.elsevier.com/locate/fitote
Short report
Antimicrobial and cytotoxic constituents from the seeds of Annona squamosa M. Mukhlesur Rahman a,*, Shahnaj Parvin a, M. Ekramul Haque a, M. Ekramul Islam a, Mohammad A. Mosaddik b a
Phytochemistry Research Laboratory, Department of Pharmacy, University of Rajshahi, Rajshahi-6205, Bangladesh b Centre for Phytochemistry, Southern Cross University, Lismore, NSW-2480, Australia Received 22 July 2003; accepted 28 April 2005
Abstract Annotemoyin-1, Annotemoyin-2, squamocin and cholesteryl glucopyranoside were isolated from the seeds of Annona squamosa. These compounds and plant extracts showed remarkable antimicrobial and cytotoxic activities. D 2005 Elsevier B.V. All rights reserved. Keywords: Annona squamosa; Antimicrobial activity; Cytotoxicity
1. Plant Annona squamosa L. (Annonaceae) [1–4] seeds were collected from Chapai Nawabganj district of Bangladesh in September 2000 and identified by Prof. A.T.M. Naderuzzaman, Department of Botany, University of Rajshahi, Bangladesh where a voucher specimen has been deposited.
* Corresponding author. Fax: +880 721 750064. E-mail address: [email protected] (M. Mukhlesur Rahman). 0367-326X/$ - see front matter D 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.fitote.2005.04.002
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489
485
2. Uses in traditional medicines Seeds, fruits and leaves are effective as insecticide, fish poison, powerful irritant of the conjunctiva and abortifacient. The root are effective as a drastic purgative and in acute dysentery [1–4].
Fig. 1. Compounds 1–4.
486
Table 1 Antibacterial activities of the A. squamosa extracts PE, CE and EE and compounds 1–4 Diameter of the zone of inhibition (mm) PE
Gram (+) B. subtilis B. cereus B. megaterium Staphylococcus aureus S. b-haemolytica Sarcina lutea Gram ( ) E. coli S. dysenteriae S. shiga S. flexneriae S sonnei Salmonella typhi P. aeruginosa Klebsiella spp.
CE
EE
1+2
3
Kanamycin
4
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
30 Ag/disc
18 20 11 12 30 18
28 21 15 14 34 22
16 15 17 12 20 22
18 17 19 14 23 26
18 16 12 8 17 18
20 19 15 11 19 22
8 10 12 13 15 10
10 11 13 15 18 12
12 13 14 15 16 13
13 15 17 18 18 16
8 8 10 10 12 10
10 10 13 13 15 13
28 30 21 40 40 30
21 12 14 13 15 18 24 23
23 15 15 15 17 20 26 25
16 10 10 12 13 13 20 18
19 12 12 13 14 16 22 21
12 12 10 11 12 14 14 14
14 14 12 11 12 16 18 18
12 14 15 11 8 13 14 18
14 15 17 13 10 16 16 21
15 12 11 13 14 17 18 21
18 13 13 16 17 19 20 22
12 15 14 8 12 15 17 14
15 18 16 11 14 17 19 16
30 15 20 18 22 25 29 25
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489
Bacteria
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489
487
3. Previous isolated class of constituents Acetogenins [5–18], terpenoids [19,20], alkaloids [21], cyclopeptides [22–25], steroids [26].
4. Tested materials Petroleum ether extract (PE), CHCl3 extract (CE), EtOH extract (EE), annotemoyin1 (1) [27], annotemoyin-2 (2) [27], squamocin (3) [28] and cholesteryl glucopyranoside (4) (Fig. 1) [29].
5. Studied activity Antibacterial and antifungal activities by disc diffusion method [30–32] and cytotoxicity by Brine shrimp bioassay [33].
6. Used microorganisms Listed in Tables 1 and 2.
7. Results All compounds as well as crude extracts showed antibacterial activities against all test organisms. Petroleum ether extract and compounds 3 and 4 did not show any antifungal activities, chloroform and ethanol extracts and compounds (1 and 2) showed reasonable activities against all test fungi except Candida albicans. The minimum inhibitory concentrations (MIC) of the compounds 1–4 were observed by serial dilution technique. The MIC value of compounds 1 and 2 was found to be 64 Ag/ml against Bacillus subtilis and Shigella shiga. The MIC values of compound 3 was 64 Ag/ml against
Table 2 Antifungal activity of the A. squamosa extracts PE, CE and EE and compounds 1–4 Fungi
Aspergillus flavus A. niger A. fumigatus C. albicans
Diameter of zone of inhibition (mm) PE
CE
EE
1+2
3
4
Nystatine
200 Ag/disc
200 Ag/disc
200 Ag/disc
200 Ag/disc
200 Ag/disc
200 Ag/disc
200 Ag/disc
– – – –
6 7 6 –
7 5 6 –
13 18 10 –
– – – –
– – – –
8 28 25 –
488
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489
Streptococcus-h-haemolyticus and Pseudomonas aeruginosa. In case of compound 4, the MIC values were 64 Ag/ml against Escherichia coli and 128 Ag/ml against Bacillus subtilis. The cytotoxicity of both extracts and compounds was studied by brine shrimp lethality bioassay. The LC50 values of petroleum ether and chloroform extracts were 0.81 and 0.83 Ag/ml, respectively. In case of compounds, the LC50 values were found to be 2.34 Ag/ml mixture of 1 and 2, 2.39 Ag/ml of 3 and 2.81 Ag/ml of 4.
Acknowledgments The authors are grateful to Prof. Naderuzzaman, Department of Botany, University of Rajshahi, Bangladesh for the identification of the plant.
References [1] Hasan MA. Amader Banaushadi Sompad. Bangladesh7 University of Dhaka; 1993. [2] Kirtikar KR, Basu BD. Indian medicinal plants. 2nd ed. India7 Lalit Mohan Basu; 1993. p. 60. [3] Yusuf M, Chowdhury JU, Wahab MA, Begum J. Medicinal plants of Bangladesh. Chittagong, Bangladesh7 BCSIR Laboratories; 1994. [4] Prain D. Bengal plants. Bot Surv India; 1963. p. 128. [5] Araya H, Sahai M, Singh S, Singh AK, Yoshida M, Hara N, et al. Phytochemistry 2002;61:999. [6] Hopp DC, Alali FQ, Gu ZM, McLaughlin JL. Bioorg Med Chem 1998;6:569. [7] Hopp DC, Zeng L, Gu ZM, Kozlowski JF, McLaughlin JL. J Nat Prod 1997;60:581. [8] Hopp DC, Alali FQ, Gu ZM, McLaughlin JL. Phytochemistry 1998;47:803. [9] Araya H, Hara N, Fujimoto Y, Sahai M. Biosci Biotechnol Biochem 1994;58:1146. [10] Hopp DC, Zeng L, Gu ZM, McLaughlin JL. J Nat Prod 1996;59:97. [11] Araya H, Hara N, Fujimoto Y, Srivastava A, Sahai M. Chem Pharm Bull 1994;42:388. [12] Fujimoto Y, Murasaki C, Shimada H, Nishioka S, Kakinuma K, Singh S, et al. Chem Pharm Bull 1994;42:1175. [13] Hirayama K, Akashi S, Yuji R, Niitsu U, Fujimoto Y. Org Mass Spectrom 1993;28:1516. [14] Fang XP, Anderson JE, Smith DL, Wood KY, Mclaughlin JL. Heterocycles 1992;34:1075. [15] Nonfon M, Lieb F, Moeschler H, Wendisch D. Phytochemistry 1990;29:1951. [16] Li XH, Hui YH, Rupprecht JK, Liu YM, Wood KV, Smith DL, et al. J Nat Prod 1990;53:81. [17] Fujimoto Y, Eguchi T, Kakinuma K, Ikekawa N, Sahai M, Gupta YK. Chem Pharm Bull 1988; 36:4806. [18] Fujimoto Y, Murasaki C, Kakinuma K, Eguchi T, Ikekawa N, Furuya M, et al. Tetrahedron Lett 1990;31:535. [19] Yang YL, Chang FR, Wu CC, Wang WY, Wu YC. J Nat Prod 2002;65:1462. [20] Wu YC, Hung YC, Chang FR, Cosentino M, Wang HK, Lee KH. J Nat Prod 1996;59:635. [21] Morita H, Sato Y, Chan KL, Choo CY, Itokawa H, Takeya K, et al. J Nat Prod 2000;63:1707. [22] Shi JX, Wu HM, He FH, Inoue K, Min ZD. Chin Chem Lett 1999;10:299. [23] Morita H, Sato Y, Kobayashi J. Tetrahedron 1999;55:7509. [24] Li CM. Phytochemistry 1998;47:1668. [25] Li CM, Tan NH, Mu Q, Zheng HL, Hao XJ, Wu Y, et al. Phytochemistry 1997;45:521. [26] Behari M, Sharma RK. J Indian Chem Soc 1986;63:255. [27] Duret P, Waechter AI, Hocquemiller R, Cave A, Batten D. Nat Prod Lett 1996;8:89. [28] Sahai M, Singh S, Singh M, Gupta YK, Akashi S, Yuji R, et al. Chem Pharm Bull 1994;42:1163. [29] Tori K, Seo S, Yoshimura Y, Arita H, Tomita Y. Tetrahedron Lett 1977;18:179.
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489 [30] [31] [32] [33]
489
Bauer AW, Kibry WMM, Sheris JC, Truck M. Am J Sci 1951; 103. Barry AL. Principle and practice of microbiology. Philadelphia7 Lea and Fabager; 1976. Reiner R. Antibiotics. An introduction. Basle-Switzerland7 Hoffman La Roche and Co; 1982. p. 70. Meyer BN, Ferrigni NR, Putnam JE, Lacobsen LB, Nichols DE, Mclaughlin JL. Planta Med 1982;45:31.
Short report
Antimicrobial and cytotoxic constituents from the seeds of Annona squamosa M. Mukhlesur Rahman a,*, Shahnaj Parvin a, M. Ekramul Haque a, M. Ekramul Islam a, Mohammad A. Mosaddik b a
Phytochemistry Research Laboratory, Department of Pharmacy, University of Rajshahi, Rajshahi-6205, Bangladesh b Centre for Phytochemistry, Southern Cross University, Lismore, NSW-2480, Australia Received 22 July 2003; accepted 28 April 2005
Abstract Annotemoyin-1, Annotemoyin-2, squamocin and cholesteryl glucopyranoside were isolated from the seeds of Annona squamosa. These compounds and plant extracts showed remarkable antimicrobial and cytotoxic activities. D 2005 Elsevier B.V. All rights reserved. Keywords: Annona squamosa; Antimicrobial activity; Cytotoxicity
1. Plant Annona squamosa L. (Annonaceae) [1–4] seeds were collected from Chapai Nawabganj district of Bangladesh in September 2000 and identified by Prof. A.T.M. Naderuzzaman, Department of Botany, University of Rajshahi, Bangladesh where a voucher specimen has been deposited.
* Corresponding author. Fax: +880 721 750064. E-mail address: [email protected] (M. Mukhlesur Rahman). 0367-326X/$ - see front matter D 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.fitote.2005.04.002
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489
485
2. Uses in traditional medicines Seeds, fruits and leaves are effective as insecticide, fish poison, powerful irritant of the conjunctiva and abortifacient. The root are effective as a drastic purgative and in acute dysentery [1–4].
Fig. 1. Compounds 1–4.
486
Table 1 Antibacterial activities of the A. squamosa extracts PE, CE and EE and compounds 1–4 Diameter of the zone of inhibition (mm) PE
Gram (+) B. subtilis B. cereus B. megaterium Staphylococcus aureus S. b-haemolytica Sarcina lutea Gram ( ) E. coli S. dysenteriae S. shiga S. flexneriae S sonnei Salmonella typhi P. aeruginosa Klebsiella spp.
CE
EE
1+2
3
Kanamycin
4
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
200 Ag/disc
400 Ag/disc
30 Ag/disc
18 20 11 12 30 18
28 21 15 14 34 22
16 15 17 12 20 22
18 17 19 14 23 26
18 16 12 8 17 18
20 19 15 11 19 22
8 10 12 13 15 10
10 11 13 15 18 12
12 13 14 15 16 13
13 15 17 18 18 16
8 8 10 10 12 10
10 10 13 13 15 13
28 30 21 40 40 30
21 12 14 13 15 18 24 23
23 15 15 15 17 20 26 25
16 10 10 12 13 13 20 18
19 12 12 13 14 16 22 21
12 12 10 11 12 14 14 14
14 14 12 11 12 16 18 18
12 14 15 11 8 13 14 18
14 15 17 13 10 16 16 21
15 12 11 13 14 17 18 21
18 13 13 16 17 19 20 22
12 15 14 8 12 15 17 14
15 18 16 11 14 17 19 16
30 15 20 18 22 25 29 25
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489
Bacteria
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489
487
3. Previous isolated class of constituents Acetogenins [5–18], terpenoids [19,20], alkaloids [21], cyclopeptides [22–25], steroids [26].
4. Tested materials Petroleum ether extract (PE), CHCl3 extract (CE), EtOH extract (EE), annotemoyin1 (1) [27], annotemoyin-2 (2) [27], squamocin (3) [28] and cholesteryl glucopyranoside (4) (Fig. 1) [29].
5. Studied activity Antibacterial and antifungal activities by disc diffusion method [30–32] and cytotoxicity by Brine shrimp bioassay [33].
6. Used microorganisms Listed in Tables 1 and 2.
7. Results All compounds as well as crude extracts showed antibacterial activities against all test organisms. Petroleum ether extract and compounds 3 and 4 did not show any antifungal activities, chloroform and ethanol extracts and compounds (1 and 2) showed reasonable activities against all test fungi except Candida albicans. The minimum inhibitory concentrations (MIC) of the compounds 1–4 were observed by serial dilution technique. The MIC value of compounds 1 and 2 was found to be 64 Ag/ml against Bacillus subtilis and Shigella shiga. The MIC values of compound 3 was 64 Ag/ml against
Table 2 Antifungal activity of the A. squamosa extracts PE, CE and EE and compounds 1–4 Fungi
Aspergillus flavus A. niger A. fumigatus C. albicans
Diameter of zone of inhibition (mm) PE
CE
EE
1+2
3
4
Nystatine
200 Ag/disc
200 Ag/disc
200 Ag/disc
200 Ag/disc
200 Ag/disc
200 Ag/disc
200 Ag/disc
– – – –
6 7 6 –
7 5 6 –
13 18 10 –
– – – –
– – – –
8 28 25 –
488
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489
Streptococcus-h-haemolyticus and Pseudomonas aeruginosa. In case of compound 4, the MIC values were 64 Ag/ml against Escherichia coli and 128 Ag/ml against Bacillus subtilis. The cytotoxicity of both extracts and compounds was studied by brine shrimp lethality bioassay. The LC50 values of petroleum ether and chloroform extracts were 0.81 and 0.83 Ag/ml, respectively. In case of compounds, the LC50 values were found to be 2.34 Ag/ml mixture of 1 and 2, 2.39 Ag/ml of 3 and 2.81 Ag/ml of 4.
Acknowledgments The authors are grateful to Prof. Naderuzzaman, Department of Botany, University of Rajshahi, Bangladesh for the identification of the plant.
References [1] Hasan MA. Amader Banaushadi Sompad. Bangladesh7 University of Dhaka; 1993. [2] Kirtikar KR, Basu BD. Indian medicinal plants. 2nd ed. India7 Lalit Mohan Basu; 1993. p. 60. [3] Yusuf M, Chowdhury JU, Wahab MA, Begum J. Medicinal plants of Bangladesh. Chittagong, Bangladesh7 BCSIR Laboratories; 1994. [4] Prain D. Bengal plants. Bot Surv India; 1963. p. 128. [5] Araya H, Sahai M, Singh S, Singh AK, Yoshida M, Hara N, et al. Phytochemistry 2002;61:999. [6] Hopp DC, Alali FQ, Gu ZM, McLaughlin JL. Bioorg Med Chem 1998;6:569. [7] Hopp DC, Zeng L, Gu ZM, Kozlowski JF, McLaughlin JL. J Nat Prod 1997;60:581. [8] Hopp DC, Alali FQ, Gu ZM, McLaughlin JL. Phytochemistry 1998;47:803. [9] Araya H, Hara N, Fujimoto Y, Sahai M. Biosci Biotechnol Biochem 1994;58:1146. [10] Hopp DC, Zeng L, Gu ZM, McLaughlin JL. J Nat Prod 1996;59:97. [11] Araya H, Hara N, Fujimoto Y, Srivastava A, Sahai M. Chem Pharm Bull 1994;42:388. [12] Fujimoto Y, Murasaki C, Shimada H, Nishioka S, Kakinuma K, Singh S, et al. Chem Pharm Bull 1994;42:1175. [13] Hirayama K, Akashi S, Yuji R, Niitsu U, Fujimoto Y. Org Mass Spectrom 1993;28:1516. [14] Fang XP, Anderson JE, Smith DL, Wood KY, Mclaughlin JL. Heterocycles 1992;34:1075. [15] Nonfon M, Lieb F, Moeschler H, Wendisch D. Phytochemistry 1990;29:1951. [16] Li XH, Hui YH, Rupprecht JK, Liu YM, Wood KV, Smith DL, et al. J Nat Prod 1990;53:81. [17] Fujimoto Y, Eguchi T, Kakinuma K, Ikekawa N, Sahai M, Gupta YK. Chem Pharm Bull 1988; 36:4806. [18] Fujimoto Y, Murasaki C, Kakinuma K, Eguchi T, Ikekawa N, Furuya M, et al. Tetrahedron Lett 1990;31:535. [19] Yang YL, Chang FR, Wu CC, Wang WY, Wu YC. J Nat Prod 2002;65:1462. [20] Wu YC, Hung YC, Chang FR, Cosentino M, Wang HK, Lee KH. J Nat Prod 1996;59:635. [21] Morita H, Sato Y, Chan KL, Choo CY, Itokawa H, Takeya K, et al. J Nat Prod 2000;63:1707. [22] Shi JX, Wu HM, He FH, Inoue K, Min ZD. Chin Chem Lett 1999;10:299. [23] Morita H, Sato Y, Kobayashi J. Tetrahedron 1999;55:7509. [24] Li CM. Phytochemistry 1998;47:1668. [25] Li CM, Tan NH, Mu Q, Zheng HL, Hao XJ, Wu Y, et al. Phytochemistry 1997;45:521. [26] Behari M, Sharma RK. J Indian Chem Soc 1986;63:255. [27] Duret P, Waechter AI, Hocquemiller R, Cave A, Batten D. Nat Prod Lett 1996;8:89. [28] Sahai M, Singh S, Singh M, Gupta YK, Akashi S, Yuji R, et al. Chem Pharm Bull 1994;42:1163. [29] Tori K, Seo S, Yoshimura Y, Arita H, Tomita Y. Tetrahedron Lett 1977;18:179.
M. Mukhlesur Rahman et al. / Fitoterapia 76 (2005) 484–489 [30] [31] [32] [33]
489
Bauer AW, Kibry WMM, Sheris JC, Truck M. Am J Sci 1951; 103. Barry AL. Principle and practice of microbiology. Philadelphia7 Lea and Fabager; 1976. Reiner R. Antibiotics. An introduction. Basle-Switzerland7 Hoffman La Roche and Co; 1982. p. 70. Meyer BN, Ferrigni NR, Putnam JE, Lacobsen LB, Nichols DE, Mclaughlin JL. Planta Med 1982;45:31.