b , Seiya Tsujimura A two-step synthesis of 7,8-dichloro ...

1

Supp

ortin

g in

form

atio

n

A tw

o-st

ep sy

nthe

sis o

f 7,8

-dic

hlor

o-rib

ofla

vin

with

hig

h yi

eld

O

livie

r Cou

rjean

a , Ann

e H

oche

deza , W

ilfrid

Ner

ia , Fré

déric

Lou

ërat

a , Em

ilie

Trem

eya , S

ébas

tien

Gou

nela , S

eiya

Tsu

jimur

ab , Nic

olas

Man

oa*

a U

nive

rsité

de

Bor

deau

x, C

entre

de

Rec

herc

he P

aul P

asca

l, C

NR

S U

PR 8

641,

115

ave

nue

Schw

eitz

er, 3

3600

Pes

sac,

Fra

nce

b Div

isio

n of

Mat

eria

ls S

cien

ce, F

acul

ty o

f Pur

e an

d A

pplie

d Sc

ienc

es, U

nive

rsity

of T

suku

ba1-

1-1

Tenn

odai

, Tsu

kuba

, Iba

raki

, 305

-857

3, Ja

pan

man

o@cr

pp-b

orde

aux.

cnrs

.fr

Exp

erim

enta

l Sec

tion.

Synt

hesi

s of t

he d

ichl

oro-

ribo

flavi

n us

ing

Boc

pro

tect

ion

(Sch

eme

1).

(1a)

: D

ichl

oro-

1,2p

heny

lene

diam

ine

(3.0

0g,

16.9

mm

ol)

was

dis

solv

ed i

n eq

uim

olar

pro

porti

on w

ith s

odiu

m b

icar

bona

te a

nd d

i-ter

t-but

yl

dica

rbon

ate

in 7

5mL

diox

ane

plus

75m

L w

ater

. The

rea

ctio

n w

as c

arrie

d ou

t for

24

hour

s at

roo

m te

mpe

ratu

re a

nd th

e re

actio

n m

ixtu

re w

as

dilu

ted

in 1

50m

L w

ater

. The

n 1a

was

ext

ract

ed tw

ice

with

50m

L di

chlo

rom

etha

ne. T

he o

rgan

ic p

hase

was

firs

t was

hed

with

50m

L sa

tura

ted

sodi

um b

icar

bona

te s

olut

ion,

then

with

100

mL

brin

e. T

he o

rgan

ic p

hase

was

drie

d us

ing

mag

nesi

um s

ulph

ate

and

evap

orat

ed u

nder

vac

uum

.

Solu

tion

(9m

L of

cyc

lohe

xan

and

3mL

of e

thyl

ace

tate

) was

add

ed to

the

brow

n so

lid a

nd fi

ltrat

ed to

giv

e th

e pu

re p

rodu

ct. Y

ield

: 2.7

4g (5

8%).

Electronic Supplementary Material (ESI) for RSC AdvancesThis journal is © The Royal Society of Chemistry 2012

2 M

ass

spec

trom

etry

ana

lysi

s (2

76.6

g m

ol-1

). 1 H

-NM

R (4

00 M

Hz,

DM

SO D

6): δ

= 8

.47

(s, 1

H),

7.53

(s, 1

H),

6.87

(s, 1

H),

5.33

(s, 2

H),

1.46

(s,

9H).

13C

-NM

R (1

00 M

Hz,

DM

SO D

6): δ

= 1

53.2

, 140

.8, 1

25.7

, 123

.9, 1

16.3

, 115

.5, 7

9.5,

66.

4, 2

8.1.

(1b)

: 1

a (1

.10g

, 4m

mol

es),

D-r

ibos

e (3

.58g

, 24

mm

oles

) an

d so

dium

cya

nobo

rohy

drid

e (0

.50g

, 16

mm

oles

) w

ere

diss

olve

d in

100

mL

drie

d

met

hano

l. Th

e re

actio

n w

as p

erfo

rmed

at 6

5°C

, for

48

hour

s. Lo

wer

am

ount

s of D

-rib

ose

and

sodi

um c

yano

boro

hydr

ide

led

to lo

wer

yie

ld o

f 1b.

Afte

r eva

pora

tion

unde

r vac

uum

, 30m

L co

ncen

trate

d aq

ueou

s H

Cl w

as u

sed

to re

act w

ith th

e re

mai

ning

sod

ium

cya

nobo

rohy

drid

e. T

he s

olut

ion

was

neu

traliz

ed w

ith sa

tura

ted

sodi

um b

icar

bona

te so

lutio

n. 1

b w

as e

xtra

cted

twic

e w

ith 5

0mL

ethy

l ace

tate

. The

org

anic

pha

se w

as w

ashe

d w

ith

brin

e, d

ried

usin

g m

agne

sium

sul

phat

e, th

en th

e so

lven

t was

rem

oved

by

evap

orat

ion

unde

r va

cuum

. Pur

ifica

tion

was

mad

e w

ith p

repa

rativ

e

chro

mat

ogra

phy

(sili

ca g

el) u

sing

AcO

Et. Y

ield

: 0.4

6g (2

8%).

Mas

s spe

ctro

met

ry a

naly

sis (

410.

8g m

ol-1

). 1 H

-NM

R (4

00 M

Hz,

DM

SO D

6) : δ

=

8.59

(s, 1

H),7

.42

(s, 1

H),

6.76

(s, 1

H),

5.32

(t, J

=4.8

1Hz,

1H

), 4.

81 (d

, J=5

.50H

z, 1

H),

4.77

(d, J

=5.5

0Hz,

1H

), 4.

68 (d

, J=5

.04H

z, 1

H),

4.42

(t,

J=5.

50H

z, 1

H),

3.78

(m, 1

H),

3.57

(m, 2

H),

3.43

(m, 2

H),

3.27

(m, 1

H),

3.02

(m, 1

H),

1.45

(s, 9

H).

13C

-NM

R (1

00 M

Hz,

DM

SO D

6): δ

= 1

53.6

,

142.

2, 1

27.1

, 125

.2, 1

24.1

, 115

.7, 1

11.2

, 79.

5, 7

3.2,

72.

8, 6

9.9,

63.

3, 4

5.6,

28.

1.

(2) :

1b

(1.1

0g, 2

.7m

mol

es) w

as d

isso

lved

in 5

3mL

of 4

M H

Cl i

n di

oxan

e. T

he re

actio

n w

as p

erfo

rmed

at r

oom

tem

pera

ture

for 5

hou

rs, u

nder

cons

tant

stir

ring.

The

sol

vent

was

rem

oved

by

evap

orat

ion

unde

r va

cuum

and

the

prod

uct w

as d

isso

lved

in w

ater

(10

0mL)

and

was

hed

thre

e

times

usi

ng 2

0mL

ethe

r. A

queo

us s

olut

ion

cont

aini

ng 2

was

free

ze, f

iltra

te a

nd d

ried

with

out f

urth

er p

urifi

catio

n. Y

ield

: m =

0.6

7g (8

0%).

Mas

s

spec

trom

etry

(ES

I) 3

10.9

g m

ol-1

. 1H

-NM

R (

400

MH

z, E

tOD

D6)

: δ

= 6.

75 (

s, 1H

), 6.

60 (

s, 1H

), 5.

42 (

s, 2H

), 3.

95 (

m, 1

H),

3.74

(m

, 2H

),


3 3.

64(m

, 2H

), 3.

51 (b

, 5H

), 3.

34 (m

, 1H

), 3.

14 (m

, 1H

). 13

C-N

MR

(100

MH

z, E

tOD

D6)

: δ =

137

.2, 1

35.4

, 121

.0, 1

19.6

, 115

.9, 1

11.8

, 73.

1, 7

0.9,

63.3

, 53.

5, 4

6.4.

(3)

: Allo

xan

(0.3

1g, 2

.2 m

mol

es)

and

boric

aci

d (0

.58g

, 9.3

mm

oles

) w

ere

diss

olve

d in

boi

ling

pure

ace

tic a

cid

(25m

L) f

or 1

hou

r, th

en th

e

solu

tion

was

coo

led

to r

oom

tem

pera

ture

. Afte

r ad

ding

0.4

2g o

f 2,

the

reac

tion

was

per

form

ed a

t 80°

C f

or 3

hou

rs a

nd, t

hen

was

left

at r

oom

tem

pera

ture

ove

rnig

ht. W

ater

(50m

L) w

as a

dded

bef

ore

evap

orat

ion

unde

r vac

uum

. Abo

ut 8

0mL

was

eva

pora

ted

and

extra

ctio

n w

as m

ade

twic

e

by 5

0mL

of c

yclo

hexa

n an

d tw

ice

by 5

0mL

by e

thyl

e ac

etat

. The

aqu

eous

pha

se w

as c

oole

d an

d a

prec

ipite

d ap

pear

. Filt

ratio

n ga

ve th

e go

od

prod

uct.

Yie

ld :

m =

0.4

3g (7

7%).

HR

MS

(ESI

) 439

.1 g

mol

-1. 1 H

-NM

R (4

00 M

Hz,

DM

SO D

6) : δ

= 11

.48

(b, 1

H),

8.36

(s, 1

H),

8.30

(s, 1

H),

5.06

(s,

1H),

4.73

(m

, 2H

), 4.

41 (

s, 1H

), 4.

13 (

s, 1H

), 3.

56 (

s, 2H

), 2.

43 (

s, 4H

). 13

C-N

MR

(10

0 M

Hz,

DM

SO D

6) : δ

= 15

9.8,

155

.8, 1

51.5

,

140.

2, 1

37.2

, 134

.5, 1

34.4

, 132

.2, 1

28.5

, 120

.1, 7

3.9,

73.

1, 6

9.1,

63.

8, 4

8.2.

Dir

ect s

ynth

esis

of d

ichl

oro-

ribo

flavi

n (S

chem

e 2)

.

D-r

ibos

e (2

.20g

, 15m

oles

), so

dium

cya

nobo

rohy

drid

e (2

.00g

, 32m

mol

es) a

nd 1

(1.7

7g, 1

0mm

oles

) wer

e di

ssol

ved

in d

ried

met

hano

l (10

0mL)

.

The

reac

tion

was

per

form

ed a

t 65°

C, f

or 4

8 ho

urs.

Low

er a

mou

nts

of D

-rib

ose

and

sodi

um c

yano

boro

hydr

ide

led

to lo

wer

yie

ld o

f 2.

Afte

r

evap

orat

ion

unde

r vac

uum

, 30m

L co

ncen

trate

d aq

ueou

s H

Cl w

as u

sed

to re

act w

ith th

e re

mai

ning

sod

ium

cya

nobo

rohy

drid

e. T

he s

olut

ion

was

neut

raliz

ed w

ith s

atur

ated

sod

ium

bic

arbo

nate

sol

utio

n. 2

was

ext

ract

ed t

wic

e w

ith 5

0mL

ethy

l ac

etat

e. T

he o

rgan

ic p

hase

was

drie

d us

ing

mag

nesi

um s

ulph

ate,

then

the

solv

ent w

as re

mov

ed b

y ev

apor

atio

n un

der v

acuu

m. T

he c

rude

pro

duct

was

par

tially

dis

solv

ed in

dic

hlor

omet

hane


4 an

d fil

tere

d on

a n

°5 fr

it. U

nsol

uble

pro

duct

was

par

tially

dis

solv

ed a

gain

with

war

m d

ichl

orom

etha

ne, c

oole

d to

room

tem

pera

ture

and

filte

red.

Yie

ld :

m =

1.0

1g (3

3%).

RM

N fo

r com

poun

d 2

as d

escr

ibed

abo

ve.

Allo

xan

(0.3

2g, 2

.2 m

mol

es)

and

boric

aci

d (0

.80g

, 12m

mol

es)

wer

e di

ssol

ved

in b

oilin

g pu

re a

cetic

aci

d (2

5mL)

for

1 h

our

and

then

cool

ed to

50°

C. A

fter a

ddin

g 0.

40g

of 2

, the

reac

tion

was

per

form

ed a

t 80°

C fo

r 3 h

ours

and

then

left

at ro

om te

mpe

ratu

re o

vern

ight

. Ext

ract

ion

and

purif

icat

ion

of 3

was

per

form

ed a

s des

crib

ed a

bove

. Yie

ld :

m =

0.4

1g (7

7%).

RM

N fo

r com

poun

d 3

as d

escr

ibed

abo

ve.

Spec

troe

lect

roch

emic

al e

xper

imen

ts

A th

in la

yer q

uartz

gla

ss s

pect

roel

ectro

chem

ical

cel

l with

a li

ght p

ass

leng

th o

f 1 m

m (A

LS) w

as u

sed

as a

n el

ectro

lysi

s ce

ll. A

Pt m

esh

(80

mes

h, 6

mm

× 7

mm

) was

use

d as

the

wor

king

ele

ctro

de. A

Pt w

ire a

nd A

n A

g|A

gCl|s

at.K

Cl (

ALS

) wer

e us

ed a

s th

e au

xilia

ry e

lect

rode

and

the

refe

renc

e el

ectro

de, r

espe

ctiv

ely.

The

se e

lect

rode

s w

ere

fixed

with

a T

eflo

n ca

p on

the

top

of th

e ce

ll. E

lect

roly

te s

olut

ion

was

0.0

67 M

phos

phat

e bu

ffer

of p

H 7

.0 c

onta

inin

g 0.

67 M

KC

l, an

d th

e to

tal v

olum

e w

as 3

00 μ

L. T

he c

once

ntra

tion

of 3

in th

e el

ectro

lyte

sol

utio

n w

as 0

.33

mg

mL-1

. The

dis

solv

ed o

xyge

n ga

s in

ele

ctro

lyte

sol

utio

n w

as re

mov

ed w

ith A

r gas

for 5

min

prio

r to

the

spec

troel

ectro

chem

ical

mea

sure

men

t

to m

inim

ize

oxyg

en le

vel.

A b

lank

et o

f A

r ga

s w

as m

aint

aine

d ov

er th

e el

ectro

lyte

sol

utio

n du

ring

the

entir

e el

ectro

chem

ical

mea

sure

men

ts.

Arg

on g

as w

as m

aint

aine

d ov

er th

e el

ectro

lyte

sol

utio

n du

ring

elec

troly

sis.

Elec

troly

sis

was

car

ried

out o

n a

pote

ntio

stat

(CH

I 660

) the

rmos

tate

d

at 1

5 ˚C

. Spe

ctru

m c

hang

e of

3 a

t eac

h po

tent

ial (

-0.6

0, -0

.50,

-0.4

5, -0

.40,

-0.3

5, -0

.30,

-0.2

5, -0

.20

and

-0.1

5 V

) was

sim

ulta

neou

sly

mon

itore

d

on a

CA

RY

100

spec

troph

otom

eter

. Th

e po

tent

ial d

epen

denc

e of

the

spec

tral c

hang

e w

as a

naly

zed

at 4

50 n

m a

ccor

ding

to fo

llow

ing

equa

tion.


5

−−

+=

O

R'

)(

)(

lnA

EA

EA

AnFRT

EE

equa

tion

(1)

whe

re n

, E, E

º’ ar

e th

e nu

mbe

r of e

lect

rons

, ele

ctro

de p

oten

tial,

and

the

redo

x po

tent

ial o

f 3, r

espe

ctiv

ely.

AR a

nd A

O a

re th

e ab

sorb

ance

at E

= –

0.6

V a

nd –

0.15

V, w

here

3 w

ould

be

fully

redu

ced

and

oxid

ized

, res

pect

ivel

y.


6 Pr

oton

NM

R sp

ectr

a of

1

Cl

Cl

NH2

NH2

H2O

DM

SO


7 abundance

01.02.03.04.05.06.07.08.09.010.011.012.013.0

X :

parts

per

Mill

ion

: Pro

ton

12.0

11.0

10.0

9.0

8.0

7.0

6.0

5.0

4.0

3.0

2.0

1.0

0-1

.0-2

.0

8.478

7.529

6.865

5.335

1.464

DM

SO d

6

9.24

2.00

1.00

0.97

0.92

Cl

Cl

NH

NH2

OO

1a

H2O


8

abundance00.010.020.030.040.050.060.070.080.090.1

X :

parts

per

Mill

ion

: Car

bon1

3

220.

021

0.0

200.

019

0.0

180.

017

0.0

160.

015

0.0

140.

013

0.0

120.

011

0.0

100.

090

.080

.070

.060

.050

.040

.030

.020

.010

.00

-10.

0-2

0.0

153.229

140.853

125.741 124.044

116.302 115.539

79.479

66.360

28.079

AcO

EtA

cOEt

AcO

Et

Cl

Cl

NH

NH2

OO

1a

DM

SO


9

abundance01.02.03.04.05.06.0

X :

parts

per

Mill

ion

: Pro

ton

12.0

11.0

10.0

9.0

8.0

7.0

6.0

5.0

4.0

3.0

2.0

1.0

0-1

.0-2

.0

8.636

7.459

6.804

5.365 4.865 4.851 4.779 4.765 4.725 4.713 4.474 4.461 4.447 3.837 3.824 3.655 3.592 3.524 3.449 3.330 3.284 3.091 3.026

9.47

2.74

2.58 2.15

1.04

0.94

0.93

0.90

0.85

0.84

0.83

0.81

Cl

Cl

NH

NH

OH

OH

OH

OH O

O1b

DM

SO

H2O


10

(thousandths)01.02.03.04.05.0

X :

parts

per

Mill

ion

: Car

bon1

3

220.

021

0.0

200.

019

0.0

180.

017

0.0

160.

015

0.0

140.

013

0.0

120.

011

0.0

100.

090

.080

.070

.060

.050

.040

.030

.020

.010

.00

-10.

0-2

0.0

159.791 155.873 151.534

140.160 137.185 134.487 134.096 132.179 128.547 120.070

73.875 73.132 69.070 63.769

48.189

Cl

Cl

NH

NH

OH

OH

OH

OH O

O1b

DM

SO


11

EtO

D

EtO

D

EtO

D


12

EtO

D

EtO

D


13

abundance01.02.03.0

X :

parts

per

Mill

ion

: Pro

ton

11.0

10.0

9.0

8.0

7.0

6.0

5.0

4.0

3.0

2.0

1.0

0-1

.0-2

.0-3

.0

10.668

7.548 7.487

4.257 4.024 3.919 3.846 3.812 3.605 3.332 2.750

1.6173.07

2.57

2.13

1.94

1.02

1.00

0.95

0.91

Cl

Cl

N N

N

NH

O

O

OH

OH

OH

OH

3

DM

SO

DM

SO


14

(thousandths)01.02.03.04.05.0

X :

parts

per

Mill

ion

: Car

bon1

3

220.

021

0.0

200.

019

0.0

180.

017

0.0

160.

015

0.0

140.

013

0.0

120.

011

0.0

100.

090

.080

.070

.060

.050

.040

.030

.020

.010

.00

-10.

0-2

0.0

159.791 155.873 151.534

140.160 137.185 134.487 134.096 132.179 128.547 120.070

73.875 73.132 69.070 63.769

48.189

Cl

Cl

N N

N

NH

O

O

OH

OH

OH

OH

3


b , Seiya Tsujimura A two-step synthesis of 7,8-dichloro ...

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