Supporting Information for
Amethysione and Amethysamide, New Metabolites from Streptosporangium
amethystogenes BCC 27081
Chollaratt Boonlarppradab, Chanwit Suriyachadkun, Sumalee Supothina and Pattiyaa
Laksanacharoen
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science
and Technology Development Agency (NSTDA), 113 Thailand Science Park, Thanon
Phahonyothin, Tambon Khlong Neung, Amphoe Khlong Luang, Pathum Thani 12120,
Thailand
Table of Contents
S1A-S1B. Table of Contents
S2. 1H NMR spectrum of amethysione (1) in acetone-d6.
S3. 13C NMR spectrum of amethysione (1) in acetone-d6.
S4. 1H-1H COSY NMR spectrum of amethysione (1) in acetone-d6.
S5. HMQC NMR spectrum of amethysione (1) in acetone-d6.
S6. HMBC NMR spectrum of amethysione (1) in acetone-d6.
S7. NOESY NMR spectrum of amethysione (1) in acetone-d6.
S8. DEPT NMR spectrum of amethysione (1) in acetone-d6.
S9. 1H NMR spectrum of amethysamide (2) in acetone-d6.
S10. 13C NMR spectrum of amethysamide (2) in acetone-d6.
S11. 1H-1H COSY NMR spectrum of amethysamide (2) in acetone-d6.
S12. HMQC NMR spectrum of amethysamide (2) in acetone-d6.
S13. HMBC NMR spectrum of amethysamide (2) in acetone-d6.
S14. NOESY NMR spectrum of amethysamide (2) in acetone-d6.
S15. DEPT NMR spectrum of amethysamide (2) in acetone-d6.
S16. 1H NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3) in acetone-d6.
S17. 13C NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3) in acetone-d6.
S18. 1H-1H COSY NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3) in
acetone-d6.
S19. HMQC NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3) in acetone-d6.
S20. HMBC NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3) in acetone-d6.
S21. NOESY NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3) in acetone-d6.
S22. DEPT NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3) in acetone-d6.
S23. 1H NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4) in
acetone-d6.
S24. 13C NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4) in
acetone-d6.
S25. 1H-1H COSY NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester
(4) in acetone-d6.
S26. HMQC NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4) in
acetone-d6.
S27. HMBC NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4) in
acetone-d6.
S28. NOESY NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4) in
acetone-d6.
S29. DEPT NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4) in
acetone-d6.
S30A-S30B. 1H NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
in acetone-d6.
S31. 13C NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5) in
acetone-d6.
S32. 1H-1H COSY NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide
(5) in acetone-d6.
S33. HMQC NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5) in
acetone-d6.
S34. HMBC NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5) in
acetone-d6.
S35. NOESY NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5) in
acetone-d6.
S36. DEPT NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5) in
acetone-d6.
S37A-S37C. Biological assays and their references
-S21
H NMR spectrum of amethysione (1) (400MHz, acetone-d6)
-S313
C NMR spectrum of amethysione (1) (400MHz, acetone-d6)
-S41
H-1H COSY NMR spectrum of amethysione (1) (400MHz, acetone-d6)
-S5HMQC NMR spectrum of amethysione (1) (400MHz, acetone-d6)
-S6HMBC NMR spectrum of amethysione (1) (400MHz, acetone-d6)
-S7NOESY NMR spectrum of amethysione (1) (400MHz, acetone-d6)
-S8DEPT NMR spectrum of amethysione (1) (400MHz, acetone-d6)
-S91
H NMR spectrum of amethysamide (2) (400MHz, acetone-d6)
-S1013
C NMR spectrum of amethysamide (2) (400MHz, acetone-d6)
-S111
H-1H COSY NMR spectrum of amethysamide (2) (400MHz, acetone-d6)
-S12HMQC NMR spectrum of amethysamide (2) (400MHz, acetone-d6)
-S13HMBC NMR spectrum of amethysamide (2) (400MHz, acetone-d6)
-S14NOESY NMR spectrum of amethysamide (2) (400MHz, acetone-d6)
-S15DEPT NMR spectrum of amethysamide (2) (400MHz, acetone-d6)
-S161
H NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3)
(500MHz, acetone-d6)
-S1713
C NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3)
(500MHz, acetone-d6)
-S181
H-1H COSY NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3)
(500MHz, acetone-d6)
-S19HMQC NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3)
(500MHz, acetone-d6)
-S20HMBC NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3)
(500MHz, acetone-d6)
-S21NOESY NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3)
(500MHz, acetone-d6)
-S22DEPT NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid (3)
(500MHz, acetone-d6)
-S231
H NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4)
(500MHz, acetone-d6)
-S2413
C NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4)
(500MHz, acetone-d6)
-S251
H-1H COSY NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4)
(500MHz, acetone-d6)
-S26HMQC NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4)
(500MHz, acetone-d6)
-S27HMBC NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4)
(500MHz, acetone-d6)
-S28NOESY NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4)
(500MHz, acetone-d6)
-S29DEPT NMR spectrum of 2-(3-phenethyl-ureido)-benzoic acid methyl ester (4)
(500MHz, acetone-d6)
-S30A1
H NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
(500MHz, acetone-d6)
-S30B1
H NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
(400MHz, acetone-d6)
-S3113
C NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
(500MHz, acetone-d6)
-S321
H-1H COSY NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
(500MHz, acetone-d6)
-S33HMQC NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
(500MHz, acetone-d6)
-S34HMBC NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
(500MHz, acetone-d6)
-S35NOESY NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
(500MHz, acetone-d6)
-S36DEPT NMR spectrum of N-phenethyl-2-(3-phenethyl-ureido)-benzamide (5)
(500MHz, acetone-d6)
Biological Assays
Cytotoxic activity against Vero cell (African green monkey kidney fibroblasts) was
evaluated by the green fluorescent protein (GFP)-based assay.1 Ellipticine was used as a
standard control for cyctotoxic assay against vero cell lines and displayed IC 50 value of
0.764 g/ml.
Antifungal activity against Candida albicans and cytotoxic screening
against cancer cell lines, including KB (human epidermoid carcinoma), MCF-7 (human
breast cancer), and NCI-H187 (human small cell lung cancer) were performed using the
resazurin microplate assay (REMA).2 The standard antifungal drug, amphotericin B,
exhibited antifungal activity with IC50 value of 0.107 g/ml, while Doxorubicin was
chosen as positive controls for cytotoxic tests against KB, MCF-7 and NCI-H187 with
respective IC50 values of 0.507 g/ml, 7.91 g/ml, and 0.071 g/ml.
Antimalarial
activity against Plasmodium falciparum K1 was derived by the microculture radioisotope
technique.3 The IC50 value of a standard antimalarial agent, dihydroartemisinin, was
0.751 ng/ml. Growth inhibition against Mycobacterium tuberculosis (H37Ra strain) was
determined by a green fluorescent protein microplate assay (GFPMA)4 and the MIC
value of a standard drug, isoniazid, was 0.0469 g/ml. Antimicrobial assay against
Enterococcus faecium, Acinetobacter baumannii and Pseudomonas aeruginosa was
performed using the optical density microplate assay (OD).5 Rifampicin, used as a
standard antibacterial drug against A. baumannii and E. faecium, exhibited MIC values at
3.13 g/ml, and 6.25-12.50 g/ml respectively. For P. aeruginosa, Erythromycin was
used as a standard control with MIC values at 8.00 g/ml. Anti-phytopathogenic fungal
activity against Magnaporthe grisea was performed using fluorometric method6 with
amphotericin B as antifungal agent, exhibited MIC value of 1.56-3.13 g/ml.
-S37A-
Measurement of activities against plant pathogens Colletotrichum capsici and
Colletotrichum gloeosporioides was carried out by fluorometric method.7 The MIC value
of a positive control amphotericin B was 1.56-3.13 g/ml. MIC and IC50 values more
than 50 g/ml were reported as inactive.
References
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(2007).
-S37B-
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Chutrakul, C., Boonruangprapa, T., Suvannakad, R., Isaka, M., Sirithunya, P.,
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Appl. Microbiol. 107, 1624–1631 (2009).
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Chutrakul, C., Khaokhajorn, P., Auncharoen, P., Boonruengprapa, T. &
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-S37C-