supporting information - wiley-vch · figure s2.the 60ms tocsy spectrum of jadomycin n (11). a) a...

Supporting Information

© Copyright Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, 2007

© Copyright Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, 2007

Supporting Information

for

On the Stereochemical Integrity of Oxazolone-Ring-Containing

Jadomycins

Charles N. Borissow, Cathy L. Graham, Ray T. Syvitski, Taryn R. Reid,

Jonathan Blay, and David L. Jakeman*

On the stereochemical integrity of oxazolone ring containing jadomycins................................. 1 Figure S1. 1H NMR data of jadomycin DM (4) and jadomycin M (9)......................................... 2 Figure S2. The 60ms TOCSY spectrum of jadomycin N (11) ................................................... 3 Methods .................................................................................................................................. 4 Cytotoxicity testing................................................................................................................... 9 Spectroscopic data................................................................................................................ 10 Table S1. Analytical mass spectroscopic data for jadomycin analogues ................................ 10 Table S2. Jadomycin DV (2) (3aS 30%)................................................................................. 11 Table S3. Jadomycin DV (2) (3aR 70%) ................................................................................ 12 Table S4. Jadomycin DT (3) (3aS 100%)............................................................................... 13 Table S5. Jadomycin DM (4) (3aS 46%) ................................................................................ 14 Table S6. Jadomycin DM (4) (3aR 54%)................................................................................ 15 Table S7. Jadomycin R-phe (5) (3aS 40%)............................................................................ 16 Table S8. Jadomycin R-phe (5) (3aR 60%)............................................................................ 17 Table S9. Jadomycin M (8) (3aS 63%)................................................................................... 18 Table S10. Jadomycin M (8) (3aR 37%) ................................................................................ 19 Table S11. Jadomycin S-phe (9) (3aR 40%).......................................................................... 20 Table S12. Jadomycin S-phe (9) (3aS 60%) .......................................................................... 21 Table S13. Jadomycin G (11) (3aX 78%) ............................................................................... 22 Table S14. Jadomycin G (11) (3aY 22%)............................................................................... 23 Table S15. Jadomycin Y (13) (3aS 65%) ............................................................................... 24 Table S16. Jadomycin Y (13) (3aR 35%)............................................................................... 25 Table S17. Jadomycin H (4) (3aR 100%)............................................................................... 26 Table S18. Jadomycin W (5) (3aS 54%) ................................................................................ 27 Table S19. Jadomycin W (5) (3aR 46%) ................................................................................ 28 Table S20. Jadomycin N (16) (3aR 100 %)............................................................................ 29 Table S21. Jadomycin bala (17) (3aX 55%) ........................................................................... 30 Table S22. Jadomycin bala (17) (3aY 45%)........................................................................... 31 References............................................................................................................................ 32

Figure S1. 1H NMR data of jadomycin DM (4) and jadomycin M (9). A) Growth using a mix-ture of D- and L-methionine (25 % and 75 % respectively) shows the same update and incor-poration rates; B) Selected areas of jadomycin DM (4); C) Selected areas of jadomycin M (9)

Figure S2. The 60ms TOCSY spectrum of jadomycin N (11). A) A unique cross peak be-

tween 3a and a resonance which does not have a cross-peak in the 13C-1H HSQC spectrum

(B) (red dashed line) indicating that the resonance at 7.89 ppm (1H) is from either an OH or

NH proton.

NH-3a

A)

B)

NH 3a 1''

Methods

All cultures of Streptomyces venezuelae ISP5320 strain VS1099 were grown as

described previously[1, 2] and by substituting the required amino acid for L-isoleucine.

Cultures were monitored at o.d.600 for cell growth and at λ526 for production of jado-

mycins. Cells were removed by vacuum filtration (Whatman #3 & 5 followed by Milli-

pore 0.45 µm type HA filter disks), and the clarified broth was passed through a 50 g

ISOLUTE Flash C18 column; the column was washed with water and adsorbed

colored components were eluted with a step-wise methanol gradient buffered with

phosphate (50 mM pH 4.0). Fractions were analyzed by TLC and relevant fractions

were combined to give crude material. Further purification was achieved using a Bio-

tage SP-1 high performance flash chromatography system (HPFC) using a Biotage

KP-C18-HS reversed-phase column and elution with a 0.1% acetic acid-acetonitrile

gradient (see supporting information online). The relevant fractions, as determined by

TLC, were concentrated and further purified using a Biotage KP-SIL normal phase

column, and eluting with dichloromethane-0.1% acetic acid in methanol. If necessary

an additional normal-phase column was performed to remove additional impurities. It

should be noted that following each purification, timely removal of the acidic solvent

(co-evaporating with n-hexanes) was critical to prevent decomposition of the jado-

mycin analogues. Purification was achieved using a Biotage SP-1 high performance

flash chromatography system (HPFC) and both Biotage KP-C18-HS reversed-phase

(RP) and Biotage KP-SIL normal phase (NP) columns using the following gradients:

Purification of jadomycin B (1): RP column, column volume (CV) = 48 mL; flow rate

= 25 mL/min; solvent A = 0.1% acetic acid in H2O; solvent B = acetonitrile; 70% A

30% B to 100% B (linear gradient) over 40 CV.

NP column, CV = 48 mL; flow rate = 25 mL/min; solvent A = dichloromethane;

solvent B = 0.1% acetic acid in methanol; 100% A to 70% A 30% B (linear gradient)

over 40 CV.


solvent B = 0.1% acetic acid in methanol; 92% A 8% B (isocratic) over 10 CV; 92% A


Purification of jadomycin DV (2): RP column, column volume (CV) = 12 mL; flow

rate = 12 mL/min; solvent A = 0.1% acetic acid in H2O; solvent B = acetonitrile; 70%

A 30% B to 100% B (linear gradient) over 40 CV.


solvent B = 0.1% acetic acid in methanol; 95% A 5% B to 80% A 20% B (linear

gradient) over 12 CV.

RP column, column volume (CV) = 12 mL; flow rate = 12 mL/min; solvent A = 0.1%

acetic acid in H2O; solvent B = acetonitrile; 70% A 30% B to 100% B (linear gradient)

over 13 CV.

Purification of jadomycin DT (3): RP column, column volume (CV) = 48 mL; flow


A 30% (isocratic) over 5 CV to 65% A 35% B (linear gradient) over 5 CV to 30% A

70% B (linear gradient) over 15 CV .


solvent B = 0.1% acetic acid in methanol; 97% A 3% B (isocratic) over 2 CV to 96%

A 4% B (linear gradient) over 3 CV; 95% A 5% B to 90% A 10% B (linear gradient)

over 12 CV.


solvent B = 0.1% acetic acid in methanol; 97% A 3% B (isocratic) over 5 CV to 91%

A 9% B (linear gradient) over 3 CV to 91% A 9% B (isocratic) for 3 CV to 60% A 40%

B (linear gradient) over 10 CV.

Purification of jadomycin DM (4): NP column, CV = 48 mL; flow rate = 25 mL/min;

solvent A = dichloromethane; solvent B = 0.1% acetic acid in methanol; 99% A 1% B

(isocratic) over 3 CV to 90% A 10% B (linear gradient) over 6 CV to 90% A 10% B

(isocratic) over 2 CV to 80% A 20% B (linear gradient) over 2 CV to 50% A 50% B

(linear gradient) over 5 CV.


acetic acid in H2O; solvent B = acetonitrile; 100% A (isocratic) over 1 CV to 100% B

(linear gradient) over 13 CV to100% A (isocratic) over 2 CV.

Purification of jadomycin R-Phe (5): RP column, column volume (CV) = 48 mL;

flow rate = 25 mL/min; solvent A = 0.1% acetic acid in H2O; solvent B = acetonitrile;

70% A 30% B to 100% B (linear gradient) over 40 CV.

NP column, CV = 48 mL; flow rate = 25 mL/min; solvent A = dichloromethane; sol-

vent B = 0.1% acetic acid in methanol; 100% A to 80% A 20% B (linear gradient)

over 40 CV.


vent B = 0.1% acetic acid in methanol; 92% A 8% B (isocratic) over 10 CV; 92% A


Purification of jadomycin V (6): RP column, column volume (CV) = 48 mL; flow rate

= 25 mL/min; solvent A = 0.1% acetic acid in H2O; solvent B = acetonitrile; 100% A to

100% B (linear gradient) over 20 CV.



over 15 CV.


acetic acid in H2O; solvent B = acetonitrile; 100% A to 40% A 60% B (linear gradient)

over 10 CV; 40% A 60% B (isocratic) over 2 CV; 40% A 60% B to 100% B (linear

gradient) over 10 CV.


vent B = 0.1% acetic acid in methanol; 90% A 10% B to 80% A 20% B (linear gradi-

ent) over 20 CV.

Purification of jadomycin T (7): RP column, column volume (CV) = 48 mL; flow rate

= 25 mL/min; solvent A = 0.1% acetic acid in H2O; solvent B = acetonitrile; 100% A to

100% B (linear gradient) over 13 CV.


vent B = 0.1% acetic acid in methanol; 98% A 2% B to 80% A 20% B (linear gradient)

over 16 CV.


acetic acid in H2O; solvent B = acetonitrile; 100% A to 100% B (linear gradient) over

10 CV.



over 10 CV.

Purification of jadomycin M (8): RP column, column volume (CV) = 12 mL; flow


A to 100% B (linear gradient) over 15 CV.



over 15 CV.



over 15 CV.

Purification of jadomycin S-Phe (9): RP column, column volume (CV) = 48 mL;

flow rate = 25 mL/min; solvent A = 0.1% acetic acid in H2O; solvent B = acetonitrile;

70% A 30% B to 100% B (linear gradient) over 40 CV.



over 40 CV.




Purification of jadomycin G (11): Initially partitioned between H2O (125 mL) and ex-

tracted with ethyl acetate (4 x 125 mL). Aqueous phase was concentrated and sub-

jected to the following purifications.

RP column, column volume (CV) = 120 mL; flow rate = 40 mL/min; solvent A = H2O;

solvent B = acetonitrile; 100% A (isocratic) over 2 CV to 70% A 30% B (linear gradi-

ent) over 8 CV.

NP column, CV = 120 mL; flow rate = 40 mL/min; solvent A = acetone; solvent B =

methanol; 95% A 5% B (isocratic) over 2 CV to 100% B (linear gradient) over 8 CV to

100 % B (isocratic) over 2 CV. The resulting fractions containing jadomycin G were

partitioned between H2O (75 mL) and extracted with ethyl acetate (6 x 75 mL) to give

the title compound.

Purification of jadomycin F (12): RP column, column volume (CV) = 48 mL; flow


A 20% B to 100% B (linear gradient) over 50 CV.


vent B = 0.1% acetic acid in methanol; 100% A to 92% A % B (linear gradient) over

40 CV. All NMR and spectroscopic details were in agreement with prior literature [3, 4]

Purification of jadomycin Y (13): RP column, column volume (CV) = 48 mL; flow


A 10% B (isocratic) 5 CV; 90% A 10% B to 80% A 20% B (linear gradient) over 4 CV;

65% A 35% B (isocratic) over 5 CV; 65% A 35% B to 50% A 50% B (linear gradient)

over 10CV.



over 10 CV.



over 40 CV.



over 40 CV.




Purification of jadomycin H (14): RP column, column volume (CV) = 48 mL; flow





over 25 CV.

Purification of jadomycin W (15): RP column, column volume (CV) = 12 mL; flow




vent B = 0.1% acetic acid in methanol; 99% A1% B to 80% A 20% B (linear gradient)

over 12 CV.

Purification of jadomyin N (16): RP column, column volume (CV) = 48 mL; flow rate =

25 mL/min; solvent A = 0.1% acetic acid in H2O; solvent B = acetonitrile; 80% A 20%

B to 100% B (linear gradient) over 50 CV.


vent B = 0.1% acetic acid in methanol; 80% A 20% B to 70% A 30% B (linear gradi-

ent) over 15 CV, 70% A 30% B to 50% A 50% B (linear gradient) over 15 CV, 50% A



vent B = 15% methanol in dichloromethane with 0.1% acetic acid; 77% A 33% B to

100% B (linear gradient) over 20 CV, then B = 0.1% acetic acid in methanol; 85% A

15% B to 100 % B (linear gradient) over 5 CV.

Purification of jadomycin β ala (17): RP column, column volume (CV) = 48 mL; flow


A 10% B (isocratic) over 1 CV to 100% B (linear gradient) over 10 CV to 100% A (iso-

cratic) over 2 CV.


vent B = 0.1% acetic acid in methanol; 100% A (isocratic) over 1 CV to 90% A 10% B

(linear gradient) over 8 CV to 90% A 10% B (isocratic) over 2 CV to 80% A 20% B

(linear gradient) over 5 CV to 80% A 20% B (isocratic) over 2 CV.


acetic acid in H2O; solvent B = 0.1% acetic acid in methanol; 100% A (isocratic) over

2 CV to 100% B (linear gradient) over 10 CV to 100% A (isocratic) over 2 CV.


acetic acid in H2O; solvent B = 0.1% acetic acid in methanol; 100% A (isocratic) over

2 CV to 20% A 80% B (linear gradient) over 10 CV to 20% A 80% B (isocratic) over 4

CV to 100% A (linear gradient) over 2 CV to 100% A (isocratic) over 5 CV.

Cytotoxicity testing: Human breast ductal carcinoma cell lines T-47D and MDA-MB-

435 (American Type Culture Collection, Manassas, VA) were cultured in the absence

of antibiotics in Dulbecco’s modification of Eagle’s medium containing 5% v/v (T-47D)

or 10% v/v (MDA-MB-435) heat-inactivated newborn calf serum. Cells (seeded at

50 000 cells/well in 96-well, flat-bottomed, polystyrene plates [Nunc]) were allowed to

adapt to culture for 48 h, and drugs were then added when the cells were at 30-45%

of confluent density. The jadomycin B analogues were dissolved in dimethyl sulfoxide

(DMSO) at 20 mM for use; after dilution all wells contained DMSO at a final concen-

tration of 0.5% v/v, which had no effect on cell viability. Following 48 h exposure to

the jadomycin B analogues, the viability of the breast carcinoma cells was evaluated

by measuring conversion of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bro-

mide to its corresponding formazan dye.[5]

Spectroscopic data

Table S1. Analytical mass spectroscopic data for jadomycin analogues.

Compound

(yield mg/L)

Growth

time (h)

Ratio of

3aR:3aS

Calc’d Mass

[M+Na]+

Expt’l

HRMS

MS/MS data

[M + H]+

[M+Na]+ Parent Ion

Aglycone Phenathro-viridin

Jadomycin B (1) (32.5)

24-48 35:65 a

Jadomycin DV (2) (1.1)

140 70:30 558.1740 558.1741 536.4 406.1 306.1

Jadomycin DT (3) (32.8)

24-48 <5 : >95 560.1533 560.1530 538.2 408.0 306.0

Jadomycin DM (4) (12.0)

24-48 55:45 590.1461 590.1465 568.1 438.0 306.0

Jadomycin R-Phe (5)

(2.5)

24-48 60:40 592.1584 592.1583 570.0 440.0 306.1

Jadomycin V (6) (10.0)

24-48 34:66 a

Jadomycin T (7) (25.0)

24-48 >95 : <5 a

Jadomycin M (8) (9.4)

24-48 37:63 590.1461 590.1463 568.1 438.1 306.1

Jadomycin S-Phe (9)

(7.2)

24-48 40:60 592.1584 592.1586 570.0 440.0 306.1

Jadomycin S (10) (7.5)

24-48 >95 : <5 a

Jadomycin G (11) (4.3)

24-48 78:22* 516.1271 516.1269 494.5 364.1 306.1

Jadomycin F(12) (12.6) a

72 45:55 a

Jadomycin Y (13) (0.9)

72 35:65 622.1689 622.1691 b

Jadomycin H (14) (1.3)

24-48 >95 : <5 618.1464 [M-H+2Na]+

618.1461 b

Jadomycin W (15) (1.3)

24-48 56:44 667.1668 667.1667 b

Jadomycin Ν (16) (6.3)

24-48 > 95 : < 5 573.1485 [M+H]+

573.1484 551.4 421.1 306.1

Jadomycin βala (17) (4.3)

24-48 55:45* 508.1608 [M+H]+

508.1605 508.2 378.1 306.1

[a] Analytical data consistent with that presented elsewhere.[3] [b] Mass spectrometry data presented previously.[6] * Due to lack of stereochemistry at C-1 of glycine and β-alanine, 3aR and 3aS were not assigned for 11 or 17.

Table S2. Jadomycin DV (2) (3aS 30%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

1'

2'

3'

UV (methanol): λmax (ε) = 211 (18532), 316 (9677), 386 (2197), 454 (1048), 516 (1445); m/z (ESI) = 557.7 [(M+Na)+, 100%], 573.7 [(M+K)+, 20%]; mass calcd for C29H29NNaO9 (M+Na)+, 558.1740; found 558.1741.

Position δ 1H [ppm] Multiplicity (J[Hz])

δ 13C [ppm]

HMBC COSY

1 5.33 d (3.5) 63.99 2´-CH3 1´ 2 170.15 3a 6.77 s 87.40 2 4 3b 4 6.77 s 114.45 3a, 6, 7a 3a, 5-CH3, 6 5 143.12

5-CH3 2.35 s 21.39 4, 5, 6 4, 6 6 6.81 s 121.05 4, 7, 7a 4, 5-CH3 7 154.12

7-OH 10.74 s 6, 7 7a 110.85 7b 8 8a 135.58 9 7.88 dd (1.5, 7.5) 122.07 11, 12a 10 10 7.84 t (8.0) 136.74 8a, 12 9, 11 11 7.63 dd (1.0, 8.5) 119.89 9, 12a 10 12 156.01

12a 117.78 13

13a 1´ 2.55 m 34.04 1, 2´-CH3, 3´-CH3

2´-CH3 1.10 d (6.5) 16.22 1, 1´, 3´-CH3 1´ 3´-CH3 1.33 d (7.0) 19.45 1, 1´, 2´-CH3 1´

1´´ 6.16 d (3.0) 94.75 3´´ 2´áx, 2´éq 2´áx 2.31 m 35.19 1´´, 2´éq, 3´´ 2´éq 2.46 ddd (1.0, 3.0, 15.0) 35.19 3´´, 4´´ 1´´, 2´áx, 3´´ 3´´ 4.02 m 65.84 2´áx, 2´éq, 4´´, 3´´-OH

3´´-OH 4.53 d (10.0) 3´´ 4´´ 3.24 dd (3.5, 9.5) 72.39 3´´, 4´´-OH, 5´´

4´´-OH 3.07 d (9.0) 4´´ 5´´ 3.77 m 66.39 4´´, 5´´-CH3

5´´-CH3 1.17 d (6.5) 17.99 4´´, 5´´ 5´´

Table S3. Jadomycin DV (2) (3aR 70%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

1'

2'

3'

Position δ 1H [ppm]

Multiplicity (J[Hz])

δ 13C [ppm]

HMBC COSY NOESY

1 5.19 d (2.5) 63.99 2´-CH3 1´ 3a 2 170.15 3a 6.27 s 87.40 2 4 1 3b 4 6.77 s 114.45 3a, 6, 7a 3a, 5-CH3, 6 5 143.12

5-CH3 2.37 s 21.39 4, 5, 6 4, 6 6 6.83 d (1.0) 121.05 4, 7, 7a 4, 5-CH3 7 154.12

7-OH 10.43 s 6, 7 7a 110.85 7b 8 8a 135.58 9 7.95 dd (1.0, 7.5) 122.07 11, 12a 10 10 7.87 t (7.5) 136.74 8a, 12 9, 11 11 7.68 dd (1.0, 8.5) 119.89 9, 12a 10 12 156.01

12a 117.78 13

13a 1´ 2.24 m 34.04 1, 2´-CH3, 3´-CH3

2´-CH3 0.51 d (7.0) 16.22 1, 1´, 3´-CH3

1´

3´-CH3 1.25 d (7.0) 19.45 1, 1´, 2´-CH3

1´

1´´ 6.12 d (3.0) 94.75 3´´ 2´áx, 2´éq 2´áx 2.31 m 35.19 1´´, 2´éq, 3´´ 2´éq 2.46 ddd (1.0, 3.0,

15.0) 35.19 3´´, 4´´ 1´´, 2´áx, 3´´

3´´ 4.02 m 65.84 2´áx, 2´éq, 4´´, 3´´-OH

3´´-OH 4.81 d (10.5) 3´´ 4´´ 3.21 dd (3.5, 10.0) 72.39 3´´, 4´´-OH, 5´´

4´´-OH 3.13 d (9.5) 4´´ 5´´ 3.77 m 66.39 4´´, 5´´-CH3

5´´-CH3 1.13 d (6.5) 17.99 4´´, 5´´ 5´´

Table S4. Jadomycin DT (3) (3aS 100%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

HO1'

UV (methanol): λmax (ε) = 212 (33895), 316 (17784), 381 (4102), 539 (2410); m/z (ESI) = 538.2 [(M+H)+, 70%], 560.0 [(M+Na)+, 100%]; MS/MS = 408.0 [aglycone, (M+H)+], 306.0 [phenanthroviridin, (M+H)+]; mass calcd for C28H27NNaO10 (M+Na)+, 560.1533; found 560.1530. Position δ 1H [ppm] Multiplicity

(J[Hz]) δ 13C [ppm]

HMBC COSY NOESY

1 4.91 d (7.5) 66.52 1´, 2, 1´-CH3

1´´, 3a 1´-CH3

2 171.24 3a 5.87 s 88.28 4 1 1´ 3b 111.97 4 6.75 s 115.00 3a, 6, 5-

CH3, 7a 5-CH3 5-CH3

5 140.54 5-CH3 2.81 s 20.14 4, 5, 6 4, 6 4, 6

6 6.71 s 119.66 4, 5-CH3, 7, 7a

5-CH3 5-CH3

7 154.04 7-OH 11.07 s 6, 7a

7a 111.48 7b 149.23 8 180.67 8a 135.73 9 7.80 d (7.5) 120.39 8, 11, 12a 10 10 7.72 t (8.0) 136.11 8a, 12 9, 11 11 11 7.48 d (8.5) 119.49 9, 12a 10 1´´, 10 12 155.20

12a 118.18 13 182.60

13a 129.52 1´ 4.54 dq (6.0) 78.50 2 1, 1´-CH3 2´áx, 3a, 1´-

CH3 1´-CH3 1.63 d (6.0) 18.96 1, 1´ 1´ 1, 1´ 1´-OH

1´´ 5.85 d (2.5) 95.53 3´´, 5´´, 12 2´áx, 2´éq 2´áx, 2´éq, 11 2´áx 2.16 dt (3.5, 15.0) 34.95 3´´, 4´´ 1´´, 2´éq, 3´´ 1´´, 3´´, 4´ 2´éq 2.32 m* 34.95 3´´, 4´´ 1´´, 2´áx, 3´´ 1´´ 3´´ 3.98 br s 66.31 1´´ 2´áx, 2´éq, 4´ 2´áx, 2´éq, 4´´,

5´´-CH3 3´´-OH

4´´ 3.15 dd (3.5, 10.5) 72.46 4´´, 5´´-CH3 3´´, 5´´ 2´áx, 5´´-CH3 4´´-OH

5´´ 3.83 dq (6.0, 10.5) 65.55 4´´, 5´´-CH3 4´´, 5´´-CH3 5´´-CH3 1.15 d (6.0) 17.34 4´´, 5´´ 5´´

*peak obscured by other signals

Table S5. Jadomycin DM (4) (3aS 46%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

S

1'

2'

UV (methanol): λmax (ε) = 206 (31877), 315 (7335), 515 (1219); m/z (ESI) = 568.1 [(M+H)+, 100%], 690.2 [(M+Na)+, 75%]; MS/MS = 438.0 [aglycone, (M+H)+], 306.0 [phenanthroviridin, (M+H)+]; mass calcd for C29H29NNaO9S (M+Na)+ 590.1461; found 590.1465.


δ 13C [ppm]

HMBC COSY NOESY

1 5.22 dd (3.5, 10.0) 58.54 2´ 1á, 1´b 1á, 1´b, 2á, 2´b 2 170.60 3a 6.53 s 87.39 1á, 1´b, 2á, 2´b,

4 3b 120.30 4 6.86 s 116.16 3a, 5-CH3, 6,

7a, 5-CH3 3a, 5-CH3

5 142.77 5-CH3 2.36 s 21.30 4, 5, 6 4, 6 4, 6

6 6.90 s 121.51 4, 5-CH3, 7, 7a

5-CH3 5-CH3, 7-OH

7 154.33 7-OH 10.57 s 6, 7 6, 9

7a 110.71 7b 143.68 8 182.44 8a 135.46 9 7.94 d (7.5) 121.68 8, 11, 12a 10 7-OH, 10 10 7.73 dd (7.5, 8.5) 136.68 8a, 12 9, 11 9, 11 11 7.45 d (8.5) 119.58 9, 12a 10 1´´, 10 12 155.49

12a 117.40 13 180.58

13a 127.28 1á 2.29 m 32.89 1, 1´b, 2á, 2´b 1´b 2.29 m 32.89 1, 1á, 2á, 2´b 2á 2.89 m 29.78 1áb, 2´b 1, 1á, 1´b 2´b 2.82 m 29.78 1áb, 2á 1, 1á, 1´b

S-CH3 2.15 s 15.96 2´ 1´´ 5.95 d (3.0) 94.47 3´´ 2´áx, 2´éq 2´áx, 2´éq, 11 2´áx 2.26 m 35.23 1´´, 2´éq, 3´´ 1´´, 3´´ 2´éq 2.54 dd (2.0, 15.0) 35.23 3´´, 4´´ 1´´, 2´áx, 3´´ 1´´, 3´´ 3´´ 4.14 m 65.92 1´´ 2´áx, 2´éq, 3´´-

OH, 4´´ 2´áx, 2´éq, 3´´-

OH, 4´´ 3´´-OH 4.69 d (9.5) 3´´ 2´áx, 2´éq, 3´´,

5´´ 4´´ 3.25 m 72.81 5´´-CH3 3´´, 4´´-OH, 5´´ 5´´, 5´´-CH3

4´´-OH 2.78 d (11.0) 4´´ 5´´ 5´´ 3.64 dq (6.0, 10.0) 66.48 4´´, 5´´-CH3 3´´-OH, 4´´, 4´´-

OH, 5´´-CH3 5´´-CH3 1.26 d (6.0) 17.97 4´´, 5´´ 5´´ 4´´, 5´´

Table S6. Jadomycin DM (4) (3aR 54%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

S

1'

2'



δ 13C [ppm]

HMBC COSY NOESY

1 5.26 dd (3.0, 6.5) 58.56 2´ 1á, 1´b 1á, 1´b, 3a 2 171.44 3a 6.15 s 87.50 1, 4 3b 120.30 4 6.84 s 114.50 3a, 5-CH3,

6, 7a, 5-CH3 3a, 5-CH3

5 143.30 5-CH3 2.37 s 21.41 4, 5, 6 4, 6 4, 6

6 6.90 s 121.08 4, 5-CH3, 7, 7a

5-CH3 5-CH3, 7-OH

7 154.34 7-OH 10.21 s 6, 7 6, 9

7a 110.80 7b 143.83 8 183.67 8a 135.54 9 7.95 d (7.5) 122.07 8, 11, 12a 10 7-OH, 10 10 7.75 dd (7.5, 8.5) 136.81 8a, 12 9, 11 9, 11 11 7.51 d (8.5) 120.14 9, 12a 10 1´´, 10 12 155.87

12a 117.66 13 181.36

13a 130.02 1á 2.19 m 30.70 1, 1´b, 2á, 2´b 1´b 2.19 m 30.70 1, 1á, 2á, 2´b 2á 2.27 m 28.81 1áb, 2´b 2´b 2.40 m 28.81 1áb, 2á

S-CH3 1.91 s 15.49 2´ 1´´ 5.91 d (3.0) 94.81 3´´ 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.20 m 35.23 1´´, 2´éq, 3´´ 2´éq 2.50 dd (2.0,

15.0) 35.23 3´´, 4´´ 1´´, 2´áx, 3´´

3´´ 4.14 m 65.97 1´´ 2´áx, 2´éq, 3´´-OH, 4´

2´áx, 2´éq, 3´´-OH, 4´´

3´´-OH 4.91 d (10.0) 3´´ 2´áx, 2´éq, 3´´, 5´´

4´´ 3.25 m 72.48 5´´-CH3 3´´, 4´´-OH, 5´´ 5´´, 5´´-CH3 4´´-OH 3.18 d (11.0) 4´´ 4´´ 5´´

5´´ 3.69 dq (6.0, 10.0)

66.40 4´´, 5´´-CH3 3´´-OH, 4´´, 4´´-OH, 5´´-

CH3 5´´-CH3 1.25 d (6.0) 17.91 4´´, 5´´ 5´´ 4´´, 5´´

Table S7. Jadomycin R-phe (5) (3aS 40%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

1'

2'

3'4'

5'

6'

UV (methanol): λmax (ε) = 207(18290), 303 (6171), 395 (1945), 484 (813), 509 (603); m/z (ESI) = 607.9 [(M+K)+, 100%]; mass calcd for C32H27NNaO9 (M+Na)+, 592.1584; found 592.1583 Position δ 1H [ppm] Multiplicity


HMBC COSY NOESY

1 6.36 s 61.92 2, 13a, 2´, 6 2´, 6´, 3´ -OH 2 169.19 3a 6.70 s 87.95 2´, 6´ 3b 4 6.91 s 115.74 5-CH3, 6, 7a 5-CH3 5-CH3 5 142.79

5-CH3 2.23 s 21.12 4, 5, 6 4, 6 4, 6 6 6.95 s 121.19 4, 5-CH3 5-CH3 5-CH3 7 154.37

7-OH 10.58 s 6, 7 6 7a 110.66 7b 8 8a 135.29 9 7.98 d (8.0) 121.86 11, 12a 10 10 10 7.72 t (8.0) 136.37 8a, 12 9, 11 9, 11 11 7.35 d (8.5) 119.03 9, 12a 10 1´´, 10 12 155.43

12a 116.68 13

13a 132.63 1´ 136.04 2´ 7.41 m 125.49 3´ 3a, 4´ -OH 3´ 7.23 m 128.94 1´ 2´, 4´ 4´´-OH 4´ 7.05 m 127.29 3´, 5´ 1 5´ 7.23 m 128.94 1´ 4´, 6´ 4´´-OH 6´ 7.41 m 125.49 5´ 3a, 4´ -OH 1´´ 5.83 d (3.5) 93.55 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.08 dt (3.5, 15.0) 34.56 1´´, 2´éq, 3´´ 1´´, 3´´ 2´éq 2.35 m* 34.56 1´´, 2´áx, 3´´ 1´´, 3´´, 3´ -OH 3´´ 3.87 m 65.24 2´áx, 2´éq,

3´´-OH, 4´´ 2´áx, 2´éq, 4´

3´´-OH 4.02 d (10.5) 3´´ 1, 5´´, 2´ eq 4´´ 3.01 dt(3.5, 10.5) 71.86 3´´, 4´´-OH,

5´´ 3´´, 4´´-OH

4´´-OH 1.90 d (10.5) 4´´ 2´, 3´, 5´, 6´, 5´´ 2.85 dq (6.0, 10.0) 65.66 4´´, 5´´-CH3 3´´-OH, 4´´-OH

5´´-CH3 1.08 d (6.0) 17.54 4´´, 5´´ 5´´ 4´´, 5´´ *peak obscured by other signals

Table S8. Jadomycin R-phe (5) (3aR 60%)

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

1'

2'

3'4'

5'

6'



δ 13C [ppm]

HMBC COSY NOESY

1 6.22 s 63.44 2, 13a, 3a 3a, 2´, 4´, 6´, 3´´-OH

2 169.61 3a 6.42 s 87.44 1, 4 3b 4 6.95 s 114.95 5-CH3, 6,

7a 5-CH3 3a, 5-CH3

5 142.40 5-CH3 2.42 s 21.26 4, 5, 6 4, 6 4, 6

6 6.98 s 121.19 4, 5-CH3 5-CH3 5-CH3 7 154.21

7-OH 10.24 s 6, 7 6, 9 7a 110.66 7b 8

8a 135.29 9 7.96 d (7.5) 121.86 11, 12a 10 10

10 7.68 t (8.0) 136.37 8a, 12 9, 11 9, 11 11 7.40 d* 119.86 9, 12a 10 1´´, 10 12 155.43 12a 117.25 13 13a 135.97 1´ 136.04 2´ 7.41 m 129.23 3´ 3a, 4´ -OH 3´ 7.23 m 128.94 1´ 2´, 4´ 4´´-OH 4´ 7.05 m 127.29 2´, 6´ 3´, 5´ 1, 3´´-OH 5´ 7.23 m 128.94 1´ 4´, 6´ 4´´-OH 6´ 7.41 m 129.23 5´ 3a, 4´ -OH 1´´ 5.87 d (3.0) 94.14 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.20 dt (3.5, 15.0) 34.93 1´´, 2´éq, 3´´ 1´´, 3´´, 4´´, 4´´-OH

2´éq 2.48 m* 34.93 1´´, 2´áx, 3´´ 1´´, 3´´, 3´ -OH 3´´ 4.09 m 65.51 2´áx, 2´éq, 3´´-

OH, 4´´ 2´áx, 2´éq, 3´´-

OH, 4´ 3´´-OH 4.73 d (10.5) 3´´ 1, 4´, 2´éq, 5´

4´´ 3.17 dt(3.0, 10.0) 71.97 3´´, 4´´-OH, 5´´ 2´áx, 3´´, 4´ -OH 4´´-OH 2.49 d (10.0) 4´´ 3´´-OH, 4´´, 5´´

5´´ 3.24 dq (6.0, 10.0)

65.93 4´´, 5´´-CH3 3´´-OH, 4´´-OH

5´´-CH3 1.08 d (6.0) 17.54 4´´, 5´´ 5´´ 4´´, 5´´ *peak obscured by other signals

Table S9. Jadomycin M (8) (3aS 63%)

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

S

1'

2'

UV (methanol): λmax (ε) = 210 (22655), 239 (11116), 317 (13600), 523 (671);m/z (ESI) = 589.9 [(M+Na)+, 100%], 643.9 [(M-H+2K)+, 65%]; mass calcd for C29H29NNaO9S (M+Na)+, 590.1461; found 590.1463.


δ 13C [ppm]

HMBC COSY

1 5.41 dd (2.5, 6.0) 58.52 2´ 1á, 1´b 2 171.39 3a 6.20 s 87.51 4 3b 120.71 4 6.84 br s 114.84 5-CH3, 6, 7a 3a, 5-CH3 5 143.40

5-CH3 2.37 s 21.41 4, 6, 7 4, 6 6 6.90 br s 121.16 4, 5-CH3, 7a 5-CH3 7 154.29

7-OH 10.04 s 6,7 7a 110.81 7b 142.58 8 183.79 8a 135.39 9 7.98 d (8.0) 121.92 8, 11, 12a 10 10 7.75 d (8.5) 136.66 8a, 12 9, 11 11 7.53 d (8.5) 120.07 9, 12a 10 12 156.00

12a 117.51 13 180.59

13a 130.04 1á 2.19 m 28.59 1, 1´b, 2á, 2´b 1´b 2.13 m 28.59 1, 1á, 2á, 2´b 2á 2.74 m 29.66 1á, 1´b, 2´b 2´b 2.24 m 29.66 1á, 1´b, 2á

S-CH3 1.89 s 15.42 1´, 2´ 1´´ 5.89 d (3.0) 94.87 3´´ 2´áx, 2´éq

2´áx 2.22 m 35.24 3´´, 4´´ 1´´, 2´éq 2´éq 2.50 dd (2.0, 15) 35.24 3´´, 4´´ 1´´, 2´áx 3´´ 4.14 m 66.04 1´´ 2´áx, 2´éq, 3´´-OH, 4´´

3´´-OH 5.01 d (10.5) 3´´ 3´´ 4´´ 3.27 d (7.5) 72.45 3´´, 4´´-OH, 5´´

4´´-OH 2.74 m 4´´ 5´´ 3.69 m 66.25 4´´, 5´´-CH3

5´´-CH3 1.27 d (6.0) 17.89 4´´, 5´´ 5´´

Table S10. Jadomycin M (8) (3aR 37%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

S

1'

2'


δ 13C [ppm]

HMBC COSY

1 5.33 dd (3.5, 8.0) 57.77 1á, 1´b 2 170.69 3a 6.50 s 87.39 4 3b 120.71 4 6.86 br s 115.69 5-CH3, 6, 7, 7a 3a, 5-CH3 5 142.84

5-CH3 2.35 s 21.30 4, 6, 7 4, 6 6 6.89 br s 121.32 4, 5-CH3, 7a 5-CH3 7 154.27

7-OH 10.68 s 6, 7 7a 110.60 7b 145.24 8 182.15 8a 135.39 9 7.92 d (8.0) 121.62 8, 11, 12a 10 10 7.71 d (8.5) 136.64 8a, 12 9, 11 11 7.47 d (8.5) 119.41 9, 12a 10 12 155.07

12a 118.22 13 181.78

13a 127.22 1´ 2.24-2.11 m 32.11 1, 2´ 2´ 2.24-2.11 m 30.17 1´

S-CH3 2.11 s 15.65 1´, 2´ 1´´ 5.85 d (3.0) 95.07 3´´ 2´áx, 2´éq

2´áx 2.22 m 35.44 3´´, 4´´ 1´´, 2´éq 2´éq 2.42 dd (2.0, 15.0) 35.44 3´´, 4´´ 1´´, 2´áx 3´´ 4.14 m 66.19 1´´ 2´áx, 2´éq, 3´´-OH, 4´´

3´´-OH 4.83 d (10.5) 3´´ 3´´ 4´´ 3.27 d (7.5) 72.42 3´´, 4´´-OH, 5´´

4´´-OH 2.74 m 4´´ 5´´ 3.79 m 66.29 4´´, 5´´-CH3

5´´-CH3 1.27 d (6.0) 17.92 4´´, 5´´ 5´´

Table S11. Jadomycin S-phe (9) (3aR 40%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b88a

910

11 12 13 13a12a

1''

2''3''4''

5 ''

H

1'

2'

3 '

4 '

5'

6 '

UV (methanol): λmax (ε) = 210 (36206), 311 (16911), 375 (4066), 516 (2620); m/z (ESI) = 607.9 [(M+K)+, 100%]; mass calcd for C32H27NNaO9 (M+Na)+, 592.1584; found 592.1586 Position δ 1H

[ppm] Multiplicity


HMBC COSY NOESY

1 6.23 s 61.50 2 3a 6.72 s 88.19 3b 4 6.92 s 115.35 7a 5-CH3 5 143.22 4, 5, 6

5-CH3 2.39 s 21.09 4, 6 6 6.95 s 121.18 5-CH3 7

7-OH 10.80 s 7a 110.47 7b 8 8a 135.08 9 7.91 dd (1.0, 8.0) 121.66 10 10 10 7.65 dd (8.0, 8.5) 136.00 8a, 12 9, 11 9, 11 11 7.42 d (8.5)* 119.61 10 10, 1´ 12 155.70

12a 117.45 13

13a 1´ 2´ 7.38 m 126.93 3´ 3´ 7.24 m 129.17 2´, 4´ 4´ 7.07 m 127.74 3´, 5´ 1 5´ 7.24 m 129.17 4´, 6´ 6´ 7.38 m 126.93 5´ 1´´ 5.70 d (2.5) 95.06 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.00 m 34.69 1´´, 2´éq, 3´´ 1´´, 2´éq, 2´éq 1.84 m 34.69 1´´, 2´áx, 3´´ 1´´, 2´áx, 3´´ 4.00 m 65.46 2´áx, 2´éq, 3´´-OH,

4´´ 4´´

3´´-OH 4.35 d (10.0) 3´´ 5´´ 4´´ 3.18 m 72.06 3´´, 4´´-OH, 5´´ 3´´, 5´´, 5´ -CH3

4´´-OH 2.67 d (10.0) 4´´ 5´´ 3.65 dq (6.0,

12.5) 65.83 4´´, 5´´-CH3 3´´-OH, 4´´, 5´´-

CH3 5´´-CH3 1.25 m* 17.69 5´´ 4´´, 5´´

*peak obscured by signals from other diastereomer

Table S12. Jadomycin S-phe (9) (3aS 60%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b88a

910

11 12 13 13a12a

1''

2''3''4''

5 ''

H

1'

2'

3 '

4 '

5'

6 '

Position δ 1H

[ppm] Multiplicity


HMBC COSY NOESY

1 6.27 s 63.50 2 3a 6.50 s 87.54 3b 4 6.97 s 115.52 7a 5-CH3 5-CH3 5 143.22 4, 5, 6

5-CH3 2.42 s 21.29 4, 6 4, 6 6 6.98 s 121.32 5-CH3 5-CH3 7

7-OH 10.15 s 7a 110.47 7b 8 8a 135.08 9 7.91 dd (1.0, 8.0) 121.66 11,

12a 10 10

10 7.65 dd (8.0, 8.5) 136.00 8a, 12 9, 11 9, 11 11 7.42 d (8.5)* 119.61 9, 12a 10 10, 1´ 12 155.70

12a 117.45 13

13a 1´ 2´ 7.38 m 129.60 3´ 1 3´ 7.24 m 129.17 2´, 4´ 4´ 7.07 m 129.05 3´, 5´ 1 5´ 7.24 m 129.17 4´, 6´ 6´ 7.38 m 129.60 5´ 1 1´´ 5.54 d (2.5) 94.67 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.14 dt (3.5, 15.5) 34.82 1´´, 2´éq, 3´´ 1´´, 2´éq, 3´´ 2´éq 2.27 m 34.82 1´´, 2´áx, 3´´ 1´´, 2´áx, 3´´, 3´ -OH 3´´ 4.15 m 65.76 2´áx, 2´éq, 3´´-

OH, 4´´ 2´áx, 2´éq, 3´´-OH

3´´-OH 4.88 d (10.5) 3´´ 2´éq, 3´´, 4´´, 5´´ 4´´ 3.25 m 72.32 3´´, 4´´-OH, 5´´ 3´´-OH, 5´´, 5´´-CH3

4´´-OH 2.77 d (10.0) 4´´ 5´´ 5´´ 3.69 dq (6.0,

12.5) 65.98 4´´, 5´´-CH3 3´´-OH, 4´´, 4´´-OH,

5´´-CH3 5´´-CH3 1.25 m* 17.69 5´´ 4´´, 5´´

*peak obscured by signals from other diastereomer.

Table S13. Jadomycin G (11) (3aX 78%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

UV (methanol): λmax (ε) = 211 (27519), 324 (15003), 532 (2277); m/z (ESI) = 515.9 [(M+Na)+, 90%]; MS/MS = 364.1 [aglycone, (M+H)+], 306.1 [phenanthroviridin, (M+H)+]; mass calcd for C26H23NNaO9 (M+Na)+ 516.1271; Found 516.1269. Position δ 1H [ppm] Multiplicity


HMBC COSY NOESY

1a 5.05 d (16.5) 57.25 3a 1b 1b 1b 4.25 d (16.5) 57.25 1a 1a, 3a 2 158.26 3a 5.18 s 63.15 1, 2, 4, 7a,

13a 1b, 4

3b 119.85 4 6.58 s 117.56 3a, 5-CH3,

6, 7a 5-CH3 3a, 5-CH3

5 139.40 5-CH3 2.29 s 19.66 4, 5, 6 4, 5, 6

6 6.68 s 117.99 4, 5-CH3, 7, 7a,

5-CH3

7 153.03 7-OH 11.56e s

7a 112.73 7b 139.02 8 179.16 8a 135.94 9 7.78 d (7.5) 119.52 8, 11, 12a 10 10 7.66 dd (8.0, 8.5) 134.69 8a, 12 9, 11 11 7.46 d (8.5) 118.84 9, 12a 10 1´´ 12 154.53

12a 118.72 13 184.40

13a 132.77 1´´ 5.79 s 95.92 3´´ 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.12 brd (15.0) 34.88 1´´, 2´éq, 3´´ 1´´, 2´éq, 3´´, 4´´

2´éq 2.43 brd (15.0) 34.88 3´´, 4´´ 1´´, 2´áx, 3´´ 1´´, 2´áx, 3´´ 3´´ 4.04 brs 66.97 1´´, 4´´ 2´áx, 2´éq, 4´ 2´áx, 2´éq, 4´ 4´´ 3.25 dd (3.0, 10.0) 72.73 5´´, 5´´-CH3 3´´, 5´´ 2´áx, 3´´, 5´´,

5´´-CH3 5´´ 3.97 dq (6.0, 10.0) 64.94 4´´, 5´´-CH3 4´´, 5´´-CH3

5´´-CH3 1.22 d (6.0) 16.81 4´´, 5´´ 5´´ 4´´, 5´´ *peak obscured by other signals. [e] exchangable.

Table S14. Jadomycin G (11) (3aY 22%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H


1a 5.17 m* 1b 4.18 d (16.5) 2 3a 5.16 s 3b 4 6.58 s 5

5-CH3 2.29 s* 6 6.68 s 7

7-OH 11.52e s 7a 7b 8 8a 9 7.82 d (7.5) 10 7.76 m* 11 7.45 m* 12

12a 13

13a 1´´ 5.93 s*

2´´ax 2.19 m* 2´´eq 2.39 m* 3´´ 4.04 m* 4´´ 3.25 m* 5´´ 3.97 m*

5´´-CH3 1.22 m* *peak obscured by signals from major diastereo-mer. [e] exchangable. 13C and 2D data are not reported as they are indistinguishable from that of 3aX due to overlapping signals.

Table S15. Jadomycin Y (13) (3aS 65%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b

88a

910

11 12 13 13a12a

1''

2''3''4 ''

5''

H

1 '

OH

3'7'

UV (methanol): λmax (ε) = 210 (4306), 283 (1400), 314 (1606), 383 (361), 516 (481); m/z (ESI) = 637.9 [(M+K)+, 100%]; mass calcd for C33H29NNaO10 (M+Na)+, 622.1689; found 622.1691.


δ 13C [ppm]

HMBC COSY NOESY

1 5.61 dd (2.5, 5.0) 61.19 1á, 1´b 1á, 1´b, 3a 2 171.05 3a 5.92 s 87.24 1, 4 3b 4 6.19 s 113.92 5-CH3, 6, 7a 5-CH3, 6 3a, 5-CH3 5 143.14

5-CH3 2.26 s 21.03 4, 5, 6 4, 6 4, 6 6 6.77 s 119.55 4, 5-CH3, 7a 4, 5-CH3 5-CH3 7 153.36

7-OH 9.95 s 6, 7 7a 110.45 7b 8 183.38 8a 135.28 9 8.01 d (7.5) 121.72 8, 11, 12a 10 10 10 7.77 t (8.0) 136.58 8a, 12 9, 11 9, 11 11 7.54 d (8.0) 119.74 9, 12a 10 1´´, 10 12 155.85

12a 117.14 13

13a 1á 3.29 m 37.89 2, 3´, 7´ 1, 1´b 1, 1´b, 3´, 7´ 1´b 3.12 dd (5.0, 14.5) 37.89 2, 3´, 7´ 1, 1á 1, 1á, 3´, 7´ 2´ 124.81 3´ 6.51 d (8.0) 130.74 1´, 5´, 7´ 4´, 6´ 1á, 1´b, 4´ 4´ 6.23 d (8.0) 114.99 2´, 6´ 3´, 7´ 3´, 4´´-OH 5´ 154.89

5´-OH 5.30 s 6´ 6.23 d (8.0) 114.99 2´, 4´ 3´, 7´ 4´´-OH, 7´ 7´ 6.51 d (8.0) 130.74 1´, 3´, 5´ 4´, 6´ 1á, 1´b, 6´ 1´´ 5.95 d (3.0) 94.75 3´´ 2´áx, 2´éq 2´áx, 2´éq, 11 2´áx 2.26 m 35.13 1´´, 2´éq, 3´´ 1´´, 2´éq, 3´´, 4´´ 2´éq 2.58 dd (2.5, 15.0) 35.13 4´´ 1´´, 2´áx, 3´´ 1´´, 2´áx, 3´´, 3´´-

OH 3´´ 4.15 m 65.97 1´´ 2´áx, 2´éq,

3´´-OH, 4´´ 2´áx, 2´éq, 3´´-OH,

4´´ 3´´-OH 5.14 d (10.0) 3´´ 2´éq, 3´´, 4´´, 5´´

4´´ 3.30 m 72.29 3´´, 4´´-OH, 5´´

2´áx, 3´´, 4´´-OH, 5´´, 5´´-CH3

4´´-OH 2.75 d (10.5) 4´´ 4´´ 5´´ 3.71 m 66.15 4´´, 5´´-CH3 3´´-OH, 4´´, 5´´-

CH3 5´´-CH3 1.26 d (6.5) 17.70 4´´, 5´´ 5´´ 4´´, 5´´

Table S16. Jadomycin Y (13) (3aR 35%).

O

O

N O

O

HOO

OOH

HO

1 2

3a

3 b4

56

7

7a7b8

8a9

10

11 12 13 13 a12a

1 ''

2 ''3''4' '

5''

H

1'

OH

3 '7'


δ 13C [ppm]

HMBC COSY NOESY

1 5.54 dd (3.0, 6.0) 60.34 1á, 1´b 1á, 1´b 2 171.05 3a 5.25 s 87.84 3´, 4, 7´ 3b 4 6.67 s 115.25 6 5-CH3, 6 3a, 5-CH3 5 142.34

5-CH3 2.29 s 21.03 4, 5, 6 4, 6 4, 6 6 6.84 s 120.95 4 4, 5-CH3 5-CH3 7 154.04

7-OH 10.58 s 6, 7 7a 110.45 7b 8 181.76 8a 135.28 9 7.97 d (7.5) 121.17 8, 11, 12a 10 10 10 7.76 t (8.0) 136.58 8a, 12 9, 11 9, 11 11 7.53 d (8.0) 119.08 9, 12a 10 1´´, 10 12 155.85

12a 117.79 13

13a 1á 3.38 dd (3.0, 14.5) 35.49 1, 1´b 1, 1´b, 3´, 7 1´b 3.25 m 35.49 1, 1á 1, 1á, 3´, 7 2´ 126.16 3´ 6.99 d (8.5) 130.74 1´, 5´, 7´ 4´, 6´ 1á, 1´b, 3a, 4´ 4´ 6.68 d (8.5) 116.26 2´ 3´, 7´ 3´, 4´´-OH 5´ 155.79

5´-OH 5.30 s 6´ 6.68 d (8.5) 116.26 2´ 3´, 7´ 4´´-OH, 7´ 7´ 6.99 d (8.5) 130.74 1´, 3´, 5´ 4´, 6´ 1á, 1´b, 3a, 6´ 1´´ 5.94 d (3.0) 94.75 3´´ 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.24 m 35.13 1´´, 2´éq, 3´´ 1´´, 2´éq, 3´´, 4´´ 2´éq 2.48 dd (2.5, 15.0) 35.13 3´´, 4´´ 1´´, 2´áx, 3´´ 1´´, 2´áx, 3´´,

3´´-OH 3´´ 4.15 m 65.97 1´´ 2´áx, 2´éq,

3´´-OH, 4´´ 2´áx, 2´éq, 3´´-

OH, 4´ 3´´-OH 4.96 d (10.5) 3´´ 2´éq, 3´´, 5´´

4´´ 3.30 m 72.29 3´´, 4´´-OH, 5´´

2´áx, 3´´, 4´ -OH, 5´´, 5´´-CH3

4´´-OH 2.75 d (10.5) 4´´ 4´´ 5´´ 3.80 m 66.15 4´´, 5´´-CH3 3´´-OH, 4´´, 5´´-

CH3 5´´-CH3 1.26 d (6.5) 17.70 4´´, 5´´ 5´´ 4´´, 5´´

Table S17. Jadomycin H (4) (3aR 100%).

O

O

N O

O

HOO

OOH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

N

NH

1'2'

6'

UV (methanol): λmax (ε) = 211 (40842), 312 (12284), 378 (3719), 469 (1697), 520 (1771); m/z (ESI) = 618.1 [(M-H+2Na)+, 100%]; mass calcd for C30H27N3Na2O9 (M-H+2Na)+, 618.1464; found 618.1461.


δ 13C [ppm]

HMBC COSY NOESY

1 6.12 d (5.0) 58.35 1á, 1´b, 3´ 2 3a 5.21 s 57.83 4 3b 4 6.74 s 117.70 5-CH3, 6 3a, 5-CH3, 6 5-CH3 5

5-CH3 2.25 s 19.85 4 4, 6 6 6.46 s 115.74 5-CH3, 6 5-CH3 7

7-OH e 7a 7b 8 181.12 8a 135.77 9 7.80 d (7.5) 119.45 8, 11, 12a 10 10 10 7.69 t (8.5) 134.57 8a, 12 9, 11 9, 11 11 7.47 d (8.5) 118.68 9, 12a 11 1´´, 10 12 153.91

12a 119.90 13

13a 1á 3.64 d (15.5) 22.50 1, 1´b, 3a 1´b 3.02 ddd (1.0, 5.5, 15.5) 22.50 1, 1á, 3´ 2´ 3´ 7.74 s 134.77 5´ 1, 1´b 4´ e 5´ note 129.42 1´´ 5.99 d (2.5) 94.32 3´´, 5´´ 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.19 dt (3.5, 15.0) 34.48 1´´, 2´éq, 3´´ 1´´, 2´éq, 2´éq 2.41 dd (1.5, 15.0) 34.48 1´´, 2´áx, 3´´ 1´´, 2´áx 3´´ 4.06 m 67.03 2´áx, 2´éq, 4´ 4´´ 3.21 dd (3.0, 10.0) 72.74 3´´, 5´´ 5´´ 4.00 dq (6.0, 10.0) 64.55 4´´, 5´´-CH3

5´´-CH3 1.19 d (6.0) 16.94 4´´, 5´´ 5´´ [e] proton not seen due to deuterium exchange

Table S18. Jadomycin W (5) (3aS 54%).

O

O

N O

O

HOO

OOH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

1'2'

NH

2'a3'

6'6'a

8'

UV (methanol): λmax (ε) = 210 (41345), 291 (14997), 319 (16852), 452 (1361), 515 (2039); m/z (ESI) = 667.1 [(M-H+2Na)+, 100%], 699.0 [(M-H+2K)+, 68%]; mass calcd for C35H29N2Na2O9 (M-H+2Na)+, 667.1668; found 667.1667.


COSY

1 5.68 dd (3.0, 5.5) 1á, 1´b 2 3a 5.96 s 3b 4 6.05 s 5-CH3, 6 5

5-CH3 2.16 s 4, 6 6 6.65 s 4, 5-CH3 7

7-OH 9.93 s 7a 7b 8 8a 9 8.02 dd (1.0, 7.5) 10 10 7.79 dd (8.0, 8.5) 9, 11 11 7.56 d (8.5) 10 12

12a 13

13a 1á 3.70 dd (5.5, 15.5) 1, 1´b 1´b 3.64 dd (2.5, 15.5) 1, 1á 2´ 2á 3´ 7.03 m 4´ 4´ 7.28* m 3´, 5´ 5´ 6.92 dd (3.5, 8.0) 4´, 6´ 6´ 7.03 m 5´ 6á 7´ 7.50 s 8´ 8´ 6.49 d (2.5) 7´ 1´´ 6.00 t (4.0) 2´áx, 2´éq 2´áx 2.32 m 1´´, 2´éq, 3´´ 2´éq 2.63 ddd (1.0, 2.0, 15.0) 1´´, 2´áx, 3´´ 3´´ 4.21 m 2´áx, 2´éq, 3´´-OH, 4´´

3´´-OH 5.17 d (10.0) 3´´ 4´´ 3.34 m 3´´, 5´´ 5´´ 3.75 m 4´´, 5´´-CH3

5´´-CH3 1.31 d (6.0) 5´´

Table S19. Jadomycin W (5) (3aR 46%).

O

O

N O

O

HOO

O

OH

HO

1 2

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

1'2'

NH

2'a3'

6'6'a

8'


COSY

1 5.66 dd (3.0, 5.5) 1á, 1´b 2 3a 5.11 s 3b 4 6.67 s 5-CH3, 6 5

5-CH3 2.30 s 4, 6 6 6.84 s 4, 5-CH3 7

7-OH 10.62 s 7a 7b 8 8a 9 8.00 dd (1.0, 7.5) 10 10 7.82 dd (8.0, 8.5) 9, 11 11 7.60 d (8.5) 10 12

12a 13

13a 1á 3.60 dd (2.5, 15.5) 1, 1´b 1´b 3.46 dd (5.5, 15.5) 1, 1á 2´ 2á 3´ 7.08 m 4´ 4´ 7.31* m 3´, 5´ 5´ 6.78 t (8.0) 4´, 6´ 6´ 7.08 m 5´ 6á 7´ 8.14 s 8´ 8´ 7.11 d (2.5) 7´ 1´´ 6.00 t (4.0) 2´áx, 2´éq

2´áx 2.27 m 1´´, 2´éq, 3´´ 2´éq 2.55 ddd (1.0, 2.0, 15.0) 1´´, 2´áx, 3´´ 3´´ 4.21 m 2´áx, 2´éq, 3´´-OH, 4´

3´´-OH 5.05 d (10.0) 3´´ 4´´ 3.34 m 3´´, 5´´ 5´´ 3.85 m 4´´, 5´´-CH3

5´´-CH3 1.33 d (6.0) 5´´

Table S20. Jadomycin N (16) (3aR 100 %).

O

O

N

HOO

OOH

HO

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

NH

OHOOC

12

3

1'

UV (methanol): λmax (ε) = 210 (18029), 315 (9255), 531 (1545); m/z (ESI) = 551.4 [(M+H)+, 100%]; MS/MS = 421.1 [aglycone, (M+H)+], 306.1 [phenanthroviridin, (M+H)+]; mass calcd for C28H26N2NaO10 (M+Na)+, 573.1485; found 573.1484. Position δ 1H

[ppm] Multiplicity


HMBC COSY NOESY

1 5.44 t (7.0) 57.91 3 2a, 2b 2a, 2b 2a 3.07 dd (7.0,

16.0) 35.47 1´ 1, 2b 1, 2b, 3a

2b 2.82 dd (7.0, 16.0)

35.47 1´ 1, 2a 1, 2a

3 166.79 NH 7.89 br s 3a 3a, 4 3a 6.05 br s 66.95 NH NH, 2a, 4 3b 4 6.74 s 117.46 3a, 5-CH3, 6,

7a 5-CH3 NH, 3a, 5-CH3

5 140.75 5-CH3 2.31 s 20.14 4, 5, 6 4, 6 4, 6

6 6.79 s 120.01 4, 5-CH3, 7a 5-CH3 5-CH3 7

7-OH 7.88 s 7a 113.60 7b 8 181.82 8a 135.09 9 7.83 d (7.5) 120.26 8, 11, 12a 10 10 7.76 dd (7.5, 8.5) 135.64 8a, 12 9, 11 11 11 7.55 d (8.5) 119.83 9, 12a 10 1´´, 11 12 154.80

12a 119.00 13 182.29

13a 1´ 167.68 1´´ 5.97 br s 95.08 3´´ 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.20 dt (3.5, 15.5) 35.02 4´´ 1´´, 2´éq, 3´´ 1´´, 2´éq, 3´´, 4´´ 2´éq 2.35 br d (15.5) 35.02 3´´, 4´´ 1´´, 2´áx, 3´´ 1´´, 2´áx, 3´´ 3´´ 3.99 m 66.31 2´áx, 2´éq,

4´´ 2´áx, 2´éq, 4´

4´´ 3.17 dd (2.5, 10.0)

72.52 5´´, 5´´-CH3 3´´, 5´´ 2´áx, 3´´, 5´ -CH3

5´´ 3.78 dq (6.0, 10.0)

65.49 4´´, 5´´-CH3 5´´-CH3

5´´-CH3 1.14 d (6.0) 17.40 4´´, 5´´ 4´´ 4´´, 5´´

Table S21. Jadomycin βala (17) (3aX 55%).

O

O

N

HOO

O

OH

HO

12

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

O

O

UV (methanol): λmax (ε) = 211 (26332), 314 (13239), 389 (2835), 526 (1868); m/z (ESI) = 530.1 [(M+Na)+, 100%]; MS/MS = 378.1 [aglycone, (M+H)+], 306.1 [phenanthroviridin, (M+H)+]; mass calcd for C27H26NO9 (M+H)+ 508.1608; found 508.1605. Position δ 1H [ppm] Multiplicity


HMBC COSY NOESY

1a 4.30 m 49.84 1b, 2a, 2b 3a 1b 4.09 m 49.84 1a, 2a, 2b 2a 3.04 m 35.31 1a, 1b, 2b 2b 2.98 m 35.31 1a, 1b, 2a 3 150.62 3a 5.53 s 92.16 1, 3, 4, 7a 1a, 4 3b 4 6.63 s 119.27 3a, 5-CH3,

6, 7a 5-CH3 3a, 5-CH3

5 140.63 5-CH3 2.36 s 21.04 4, 5, 6 4, 6 4, 6

6 6.92 s 120.46 4, 5-CH3, 7a

5-CH3 5-CH3

7 154.12 7-OH 10.90 s 6, 7

7a 112.89 7b 8 182.60 8a 135.43 9 7.90 d (7.5) 121.14 8, 11, 12a 10 10 7.67 dd (8.0, 8.5) 135.76 8a, 12 9, 11 11 7.43 d (8.5) 119.23 9, 12a 10 1´´ 12 154.12

12a 118.97 13 184.11

13a 1´´ 5.88 br s 94.53 3´´ 2´áx, 2´éq 2´áx, 2´éq, 11

2´áx 2.21 dt (3.5, 15.0) 35.02 4´´ 1´´, 2´éq, 3´´ 2´éq, 3´ 2´éq 2.48 br d (15.0) 35.02 1´´, 2´áx, 3´´ 2´áx, 3´´ 3´´ 4.23 m 66.86 1´´ 2´áx, 2´éq, 4´ 2´áx, 2´éq, 4´ 4´´ 3.29 dq (3.5, 10.0) 72.41 5´´-CH3 3´´, 5´´ 3´´, 5´´-CH3 5´´ 3.82 dq (6.0, 10.0) 65.96 4´´, 5´´-CH3 5´´-CH3

5´´-CH3 1.28 d (6.0) 17.78 4´´, 5´´ 5´´ 4´´, 5´´

Table S22. Jadomycin βala (17) (3aY 45%).

O

O

N

HOO

O

OH

HO

12

3a

3b4

56

7

7a7b8

8a9

10

11 12 13 13a12a

1''

2''3''4''

5''

H

O

O

Position δ 1H [ppm] Multiplicity


HMBC COSY NOESY

1a 4.15 m* 49.84 1b, 2a, 2b 1b 3.97 m 49.84 1a, 2a, 2b 2a 2.87 m 35.31 1a, 1b, 2b 2b 2.84 m 35.31 1a, 1b, 2a 3 149.28 3a 5.42 s 91.59 1, 3, 4, 7a 4 3b 4 6.66 s 118.48 3a, 5-CH3,

6, 7a 5-CH3 3a,

5 140.97 5-CH3 2.39 s 21.07 4, 5, 6 4, 6

6 6.95 s 120.56 4, 5-CH3, 7a

5-CH3

7 154.15 7-OH 10.67 s 6

7a 112.79 7b 8 182.46 8a 135.45 9 7.88 d (7.5) 121.33 8 10 10 7.66 t (8.0) 135.70 8a, 12 9, 11 11 7.41 d (8.5) 118.87 9, 12a 10 12 154.11

12a 118.95 13 183.82

13a 1´´ 5.88 br s 94.31 3´´ 2´áx, 2´éq

2´áx 2.12 dt (3.5, 15.0) 34.78 1´´, 2´ eq, 3´´ 2´éq 2.33 br d (15.0) 34.78 4´´ 1´´, 2´áx, 3´´ 3´´ 4.16 m 66.49 1´´ 2´áx, 2´éq, 4´ 4´´ 3.29 dq (3.5, 10.0) 72.36 5´´-CH3 3´´, 5´´ 5´´ 3.72 dq (6.0, 10.0) 66.07 4´´, 5´´-CH3

5´´-CH3 1.26 d (6.0) 17.80 4´´, 5´´ 5´´ 5´´ *peak obscured by signals from major diastereomer

References

[1] D. L. Jakeman, C. L. Graham, W. Young, L. C. Vining J. Ind. Microbiol. Biotechnol. 2006,

33, 767-772.

[2] R. T. Syvitski, C. N. Borissow, C. L. Graham, D. L. Jakeman Org. Lett. 2006, 8, 697-700.

[3] U. Rix, J. Zheng, L. L. Remsing Rix, L. Greenwell, K. Yang, J. Rohr J. Am. Chem. Soc.

2004, 126, 4496-4497.

[4] J. L. Doull, S. W. Ayer, A. K. Singh, P. Thibault J. Antibiot. 1993, 46, 869-871.

[5] J. Blay, W. M. MacKenzie, D. W. Hoskin In Vitro Cell. Dev. Biol. 1997, 33, 731-733.

[6] D. L. Jakeman, S. Farrell, W. Young, R. J. Doucet, S. C. Timmons Bioorg. Med. Chem.

Lett. 2005, 15, 1447-1449.

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