CHEMICAL CONSTITUENTS FROM THE STEM BARK OF <i>Annona pickelii</i> (Annonaceae)

Costa, Emmanoel Vilaça; Sampaio, Marília Fernanda Chaves; Salvador, Marcos José; Nepel, Angelita; Barison, Andersson

doi:10.5935/0100-4042.20150069

Emmanoel Vilaça Costa ^**e-mail: emmanoelvc@gmail.com

Departamento de Química, Universidade Federal de Sergipe, Avenida Vereador Olímpio Grande, S/N, Campus Prof. Alberto Carvalho, 49500-000 Itabaiana - SE, Brasil

Departamento de Química, Universidade Federal do Amazonas, Avenida General Rodrigo Otavio Jordão Ramos, 6200, Campus Sen. Arthur Virgílio Filho, 69077-000 Manaus - AM, Brasil

Marília Fernanda Chaves Sampaio

Departamento de Química, Universidade Federal de Sergipe, Av. Marechal Rondon, S/N, Cidade Universitária Prof. José Aloísio de Campos, 49100-000 São Cristóvão - SE, Brasil

Marcos José Salvador

Departamento de Biologia Vegetal, Instituto de Biologia, Universidade Estadual de Campinas, CP 6109, 13083-970 Campinas - SP, Brasil

Angelita Nepel

Departamento de Química, Universidade Federal do Paraná, Centro Politécnico, CP 19081, 81531-990 Curitiba - PR, Brasil

Andersson Barison

Departamento de Química, Universidade Federal do Paraná, Centro Politécnico, CP 19081, 81531-990 Curitiba - PR, Brasil

*e-mail: emmanoelvc@gmail.com

Position	Anonaine (11)		Asimilobine (12)		Analobine (13)
Position	δ_C mult.^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 ,^c c Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	δ_H mult. (J in Hz)^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3	δ_C mult.^b b or CDCl3 with some drops of CD3OD. ,^c c Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	Sh mult. (J in Hz)^b b or CDCl3 with some drops of CD3OD.	δ_C mult.^b b or CDCl3 with some drops of CD3OD. ,^c c Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	δ_H mult. (J in Hz)^b b or CDCl3 with some drops of CD3OD.
1	142.5, C		143.5, C		143.0, C
1a	116.1, C		125.9, C		115.7, C
2	146.8, C		148.9, C		148.3, C
3	107.9, CH	6.57 s	115.1, CH	6.68 s	107.6, CH	6.50 s
3a	126.4, C		129.2, C		127.2, C
3b	128.1, C		127.4, C		127.3, C
4_pseudoeq	29.0, CH₂	3.01 m	28.3, CH₂	3.01 m	29.4, CH₂	2.67 m
4_pseudoax		2.67 m		2.70 m		2.30 m
5_pseudoeq	43.1, CH₂	3.41 m	42.8, CH₂	3.35 m	43.9, CH₂	3.33 m
5_pseudoax		3.02 m		3.02 m		2.96 m
6a	53.3, CH	3.99 dd (14.2; 4.9)	53.4, CH	3.81 dd (13.8; 4.6)	54.5, CH	3.89 m
7_pseudoeq	36.8, CH₂	2.96 dd (14.2; 4.9)	36.9, CH₂	2.86 dd (13.8; 4.6)	37.4, CH₂	2.84 m
7_pseudoax		2.83 dd (14.2; 14.2)		2.76 brd (13.8)		2.74 m
7a	134.9, C		135.7, C		137.6, C
8	128.1, CH	7.25 m	127.3, CH	7.31 ddd (6.7, 2.0, 0.6)	115.8, CH	6.70 d (2.7)
9	127.5, CH	7.22 m	127.6, CH	7.22 m	158.0, C
10	127.00 CH	7.31 m	128.0, CH	7.22 m	114.9, CH	6.71 dd (8.3, 2.7)
11	127.03, CH	8.07 brd (7.7)	127.6, CH	8.32 dd (8.1, 0.6)	129.5, CH	7.91 d (8.3)
11a	131.1, C		132.9, C		123.9, C
1-OCH₂O-2	100.6, CH₂	5.94 d (1.4)			101.7, CH₂	5.90 d (1.2)
		6.09 d (1.4)				6.07 d (1.2)
H₃CO-1			60.2, CH₃	3.60 s

Position	Discretamine (14)			Stepholidine (15)
Position	δ_C mult.^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD. ,^b b Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	δ_H mult. (J in Hz)^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD.	NOE	δ_C mult.^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD. ,^b b Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	δ_H mult. (J in Hz)^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD.	NOE
1	108.3, CH	6.71 s	H₃CO-2; H- 13_pgeudoeq	112.0, CH	6.77 s
2	145.O, C			144.6, C
3	144.4, C			146.3, C
4	114.7, CH	6.64 s	H-5_pseudoeq	111.4, CH	6.61 s	H₃CO-3; H-5_pseudoeq
4a	127.0, C			125.5, C
5_pseudoeq	28.4, CH₂	3.08 ddd (17.0; 12.0;5.0)		28.7, CH₂	3.11 m
5_pseudoax		2.67 m			2.69 m
6_pseudoeq	51.6, CH₂	3.19 ddd (10.9; 5.0;1.6)		52.0, CH₂	3.21 m
6_pseudoax		2.63 m (11.9; 10.8; 2.9)			2.67 m
8_pseudoeq	53.9, CH₂	4.20 d (15.5)	H-8_pseudoax	54.1, CH₂	4.20 d (15.6)	H-8_pseudoax
8_pseudoax		3.56 d (15.5)			3.55 d (15.6)
8a	127.8, C			127.9, C
9	143.6, C			143.8, C
10	147.2, C			147.5, C
11	115.1, CH	6.77 d (8.2)		115.4, CH	6.76 d (8.3)
12	124.7, CH	6.81 d (8.2)		124.7, CH	6.80 d (8.3)
12a	126.5, C			126.4, C
13_pseudoeq	35.9, CH₂	3.28 dd (16.0; 3.8)		35.8, CH₂	3.28 dd (16.0; 3.9)
13_pseudoax		2.82 dd (16.0; 11.4)			2.78 dd (16.0; 11.4)
13a	59.6, CH	3.59 dd (11.4; 3.8)		59.6, CH	3.57 dd (11.4; 3.9)
13b	128.7, C			129.9, C
H₃CO-2	56.2, CH₃	3.88 s	H-1
H₃CO-3				56.0, CH₃	3.86 s	H-4
H₃CO-9	60.3, CH₃	3.82 s	H-8_pseudoeq	60.2, CH₃	3.83 s	H-8_pseudoeq

Position	Coclaurine (16)		Orientaline (17)		Juziphine (18)
Position	δ_C mult.^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD. ,^b b Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	δ_H mult. (J in Hz)^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD.	δ_>C mult.^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD. ,^b b Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	δ_H mult. (J in Hz)^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD.	δ_C mult.^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD. ,^b b Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	δ_H mult. (J in Hz)^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3 + drops of CD3OD.
1	57.8, CH	4.13 dd (9.4; 4.4)	65.8, CH	3.82 m	61.9, CH	4.39 dd (8.9; 2.8)
3_pseudoeq	41.2, CH₂	3.22 m	47.4, CH₂	3.22 m	46.0, CH₂	3.31 m
3_pseudoax		2.93 m		2.84 m		2.82 m
4_pseudoeq	28.8, CH₂	2.78 m	25.5, CH₂	2.88 m	24.2, CH₂	2.91 m
4_pseudoax		2.78 m		2.73 m		2.63 m
4a	126.1, C		124.6, C		126.5, C
5	112.7, CH	6.65 s	112.4, CH	6.64 s	120.1, CH	6.61 d (8.4)
6	148.0, C		148.0, C		111.4, CH	6.83 d (8.4)
7	145.7, C		145.2, C		146.6, C
8	114.0, CH	6.69 s	115.5, CH	6.14 s	144.2, C
8a	129.6, C		129.3, C		124.0, C
9	41.7, CH₂	3.17 dd (14.2, 4.4)	40.9, CH₂	3.07 m	38.8, CH₂	3.08 m
		2.84 dd (14.2, 9.4)		2.77 m		2.93 m
10	128.8, C		132.9, C		132.9, C
11	131.3, CH	7.07 d (8.5)	121.8, CH	6.54 dd (8.2; 2.1)	131.2, CH	7.11 d (8.6)
12	116.6, CH	6.77 d (8.5)	112.7, CH	6.80 d (8.2)	116.0, CH	6.70 d (8.6)
13	157.2, C		147.3, C		156.6, C
14	116.6, CH	6.77 d (8.5)	147.5, C		116.0, CH	6.70 d (8.6)
15	131.3, CH	7.07 d (8.5)	117.4, CH	6.63 d (2.1)	131.2, CH	7.11 d (8.6)
H₃CO-6	56.4, CH₃	3.83 s	56.3, CH₃	3.81 s
H₃CO-7					56.6, CH₃	3.86 s
H₃CO-14			56.4, CH₃	3.83 s
h₃c-n			42.3, CH₃	2.54 s	42.7, CH₃	2.43 s

Position		Stepharine (19)
	δ_C mult.^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3. ,^b b Multiplicities determined by DEPT 135, HSQC e HMBC experiments.	δ_H mult. (J in Hz)^a a The experiments were acquired at 293 K with TMS as internal reference (0.00 ppm) in CDCl3.
1	144.3, C
2	153.4, C
3	112.7, CH	6.66 s
3a	128.8, C
4_pseudoeq	26.1, CH₂	2.79 m
4_pseudoax		2.79 m
5_pseudoeq	44.8, CH₂	3.44 ddd (12.7; 6.6; 2.3)
5_pseudoax		3.14 ddd (12.7; 10.3; 6.6)
6a	57.9, CH	4.30 dd (10.8; 6.3)
6b	134.7, C
7_pseudoeq	48.2, CH₂	2.41 dd (12.1; 6.3)
7_pseudoax		2.21 dd (12.1; 10.8)
8	150.8, CH	7.06 dd (10.0; 2.9 )
9	127.6, CH	6.30 dd (10.0; 1.9)
10	186.7, C
11	128.4, CH	6.42 dd (10.0; 1.9)
12	154.3, CH	6.93 dd (10.0; 2.9)
12a	51.4, C
12b	137.0, C
H₃CO-1	61.4, CH₃	3.60 s
H₃CO-2	56.6, CH₃	3.81 s

Microorganisms	Antifungal and antibacterial activities expressed as MIC^a a MIC (minimum inhibitory concentration with 100% of microorganism inhibition) in μg mL-1; (μg mL^-1)
Microorganisms	Hexane extract	Methanolic extract	Alkaloidal fraction	Neutral fraction	Eudesmin (5)	Magnolin (6)	Yangambin (7)	Discretamine (14)	Stepholidine (15)	Positive controls^b b positive controls (chloramphenicol for bacteria strains and ketoconazole for yeast strains);
Bacillus subtilis (Bs)^d d field strain.	250	250	500	500	-	-	-	-	-	50
Candida albicans (ATCC 10231)^c c standard strain; dfield strain.	5000	5000	500	1000	125	125	125	>500	>500	12.5
Candida albicans (ATCC 1023)^c c standard strain; dfield strain.	5000	5000	1000	1000	125	125	125	>500	-	12.5
Candida parapsilosis (ATCC 22019)^c c standard strain; dfield strain.	5000	500	500	500	-	-	-	125	-	12.5
Candida tropicalis (ATCC 157)^c c standard strain; dfield strain.	5000	5000	5000	1000	250	500	-	>500	-	12.5
Candida tropicalis (CT)^d d field strain.	5000	5000	5000	1000	250	-	-	>500	-	12.5
Candida glabrata (ATCC 30070)^c c standard strain; dfield strain.	5000	5000	250	1000	-	-	-	-	-	12.5
Candida dubliniensis (ATCC 778157)^c c standard strain; dfield strain.	5000	5000	1000	1000	250	250	-	125	-	12.5
Enterobacter aerogenes (Ea)^d d field strain.	500	500	250	500	-	-	-	-	-	50
Escherichia coli (ATCC 10538)^c c standard strain; dfield strain.	> 5000	> 5000	1000	> 5000	-	-	-	-	-	50
Escherichia coli (ATCC 10799)^c c standard strain; dfield strain.	1000	1000	1000	1000	-	-	-	-	-	50
Proteus vulgaris (Pv)^d d field strain.	500	1000	500	500	500	-	-	>500	-	50
Pseudomonas aeruginosa (ATCC 27853)^c c standard strain; dfield strain.	> 5000	1000	1000	1000	-	-	-	-	-	850
Staphylococcus aureus (ATCC 14458)^c c standard strain; dfield strain.	500	500	500	500	500	500	500	-	-	25
Staphylococcus aureus (ATCC 6538)^c c standard strain; dfield strain.	> 5000	> 5000	500	> 5000	500	500	500	-	-	25
Staphylococcus epidermidis (ATCC 1228)^c c standard strain; dfield strain.	1000	500	1000	500	-	-	500	>500	-	50
Staphylococcus epidermidis (6epi)^d d field strain.	1000	500	1000	500	-	-	500	>500	-	50
Antioxidant activity (ORAC-FL)
	352.73	1925.82	4545.04	862.60	-	-	-	2.10 (1.35)^f f ORAC data expressed as Trolox equivalent relative, mean (%RSD, relative standard deviation) of triplicate assays;	0.78 (1.57)^f f ORAC data expressed as Trolox equivalent relative, mean (%RSD, relative standard deviation) of triplicate assays;	^* * Positive controls (Caffeic acid: 2.85 (1.15)f, quercetin: 5.60 (1.20)f and isoquercitrin: 5.10 (1.40)f).
	(3.60)^e e ORAC data expressed as mmol of TE g-1 of extract or fraction, mean (%RSD, relative standard deviation) of triplicate assays; μ	(1.37)^e e ORAC data expressed as mmol of TE g-1 of extract or fraction, mean (%RSD, relative standard deviation) of triplicate assays;	(2.46)^e e ORAC data expressed as mmol of TE g-1 of extract or fraction, mean (%RSD, relative standard deviation) of triplicate assays;	(7.01)^e e ORAC data expressed as mmol of TE g-1 of extract or fraction, mean (%RSD, relative standard deviation) of triplicate assays;

[1] *e-mail: emmanoelvc@gmail.com

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CHEMICAL CONSTITUENTS FROM THE STEM BARK OF Annona pickelii (Annonaceae)