The total synthesis of myo-inositol polyphosphates

Tetrahedron Vol. 45, No. Printed in Great Britain.

17. pp.

5679

to 5702,

1989 0

COW4@20/89 S3.00+ .@I 1989 Pergamon Press plc

THE TOTAL SYNTHESIS OF myo_INOSlTOL POLYPHOSPHATES

Jo-h

P. Veuxa,*’ S. Jsne deSolms,l Joel R. Huff,’ DewldC. Bllllngton,*2 Raymond Saker,2

Janusz J. KU~OQOWM 2 and lan M. Mawer 2

‘Msrokshalp&Dohme-

westPow

PsfNlsylwls 194e6,2Msrckshalp Centm,TedIn@ park. EasNck Road,

Laboratories,

&oohme--,NeWWWICeHadow,EssexCM20 MR. U.K.

(Received in UK 19 April 1989)

Abstract TomI syntheaIs oftha MM&al enantkmen, ofnlyo-lnosllolcphoaphats(g, rnyo-lnosltol 1&blspho@ste (2) and myrArMtoll,4,5-thpho~phate (lJ, tog6thw with syof rwemk myo-lnosltof l,3J+sphorphate &I and myo+o8tlol2,4,5-mphate (5J am reported. The s~eofeaRnetheuM,otcemphenkaddeotsn,for~lutlonotprotectsdI~~ols,endtheuse of teWNlKyIpyrophoapIlate aa ml eftkIenl phot3ptWyIaung egentfor polyllydmy akQhol5.

Recently hydrolysis

it has been

of

found

D-myo-inositol-1,4,5_trisphosphate directly

mediates

the action

agonist

[1,4,5-IP3,

the release

of 1,4,5-IP3

5-phosphatase

that

stimulation

phosphatidylinositol-4,5_bisphosphate of calcium

is believed

present

in the

further

degraded

inositol

which is recycled

by other

1,4,5-IP3 degradation

first

recycled

adequate

supply of the natural inositol

phosphates

which

A key problem

encountered

suitably

protected

myo-inositol

inositol

will deal

ketals with

to the

natural

subsequent products

@I,

[2,4,5-IP3,

related which

us to develop of inositol

A popular

1, 5, and 9 in large

5679

been

amount quantities

inositol

is the synthesis problem

of acid,

as a by-

pathway, available

of ketals

for

and resolution all three

I).

an from

method

involves

giving

lob (Scheme

groups

occurring

calcium.2

are not readily

to this

for

group of

metabolized

insolated

and efficient in preliminary form. 899

of protecting

4 2, 2, 2 and g.

3-hvdroxyl

is

free

pathway

to the naturally

to the

phosphates

(?)I,

and ultimately

a general

solution

and a catalytic

manipulation

[1,4-IP2,

An alternate

to release

processes

and

for terminating

formed,

has

and

group by a specific

of the

which

has also been shown

in the synthesis

of myo_inositol the

The major pathway

In addition

in the

messenger

to 1,3,4-IP3 (3) which is further

we have communicated

derivatives.

as a second

phosphorylation

IP pathway.

led

results

diacylglycerol’

of the 5-phosphate

as well as derivatives

need

(5) with ethyoxycyclohexeneloa

biscyclohexene

conversion

was

involves

biochemical

products, This

sources

synthesizing

paper

the fundamental

needed.

natural

of

to study

acts

more (Ptd)Ins(4,5)P2.4Y5

in the

phosphates, D-my2,4,5_trisphosphate product 277 in the preparation of 1,4,5-IP3,

give

The 1,4-bisphosphate

(21. This is dephosphorylated

are then

inositol

In order

stores.2

of receptors

to

to give I- and 4-monophosphates

which

identified5

1,4,5-IP3

removal

membranes.3

phosphatases

has been

that

plasma

through

in the brain to provide

1,4,5-IP3 to give 1,3,4,5-IP4 to bisphosphates

Q) Fig I].**~

from intracellular

to occur

of a number

[(Ptd)Ins(4,5)P21

treating of the

The following

-7 and S and their

J. P. VACCA et al.

5680

FIGURE 1

OH H2OsPO

H202”O 0*2H2

OH

OPO2H2

0

@I

0

OH OH

OH OH 0

OPO&?

OH

@I

0

SCHEME 1 OEt

,

1

( p-TsOH,DMF,100°C,2h.

,

OH

7 38%

898%

9 19%

5681

myo-Inositol polyphosphates

SCHEME 2

Ester =

b

10

Ester

R=Bn

H PO(OPh)2

Q Q 0

0

RO

OH

O&l

e”

9

o

D

OH

-6 140

Scheme

(a)

P = PD,Bn,

15D

4-IP

2

NM,

PhCH2Br,

25’~; (c) KOH, EtOH, 2.5’~; (f) 1096 W/C,

25’~;

PhMe,

reflux;

(b) S(-)-camphanic

(d) @hO)2POCI,

EtOH-HZ0

(80:20),

CH2CL2,

H2, 50 psig,

(Et$N,

25”~.

acid chloride, DMAP,

25’~;

CH2CL2,

(E$N,

(e) PhCH20H,

DMAP,

NaH, THF,

5682

J. P. VACCA et

al.

RCSUJt.5 D- and L-~inositol (8) was alkylated with

4-Jhosphate: (Scheme

selectively

St-l-camphanic

quantitative followed from

yield.

HPLC

myo-inositol

to give

polar

The resultant

diastereomeric 1:2).

Silica 60).

The use of camphanic

acid

alcohol

camphanate

was obtained

(EtOAc/petrol,

by MPLC (Lichoprep

derivatives

1,2:4,5-di-O-cyclohexylidene

in 60% yield. the

diastereomer

recrystallizations

liquors

and ‘H NMR.

resolving

chloride

The more

by two more

the mother

in the 3-position

acid

2) Racemic

provides

than that of mono-glycosides

-JJD and -11L in from the mixture

diastereomer

was recovered

showed >99% purity

Both diastereomers

esters

g was acylated

esters

by crystallization

The second

myo-inositol

a more

convenient

which was first pioneered

by

method

for by Stepanov. 11

alcohols -12D ([a], = -26.1’) and 12L ([aID = 25.99. The alcohol 12D was phosphorylated in the normal manner to yield the diphenyl protected inositol phosphate, !3&. This was transesterified using the anion of benzyl alcohol in

Basic hydrolysis

of the camphanate

tetrahydrofuran

(THF)

deprotection inositol

$-phosphate

isomeric

to

give

of the benzyl

inositol

employing

the

afforded

dibenzyl

groups followed

15D, which phosphates

the same

esters

methodology

evident yielded

synthesized

from

resolved

diastereomers.’ we found chloride

ester

biscamphanate esters

dials -17D ([aID = 15.70) and & with diphenylphosphoryl

groups

D-

of each

protected

as well as cleavage

and L-~inositol

to develop

the

to report

first

gJ3 .

0zakiJ4

has

major

expeditiously product

a syntheses

method

= 1.1’).

by tedious

a similar

in four

ketalization

These were separated

([a],

separation

of the

resulted

in deprotection

ketals ([al,

(Scheme

synthesis

of myo_inositol

monoesters

afforded

yield. of the

esters

g.

protecting

1,4-bisphosphate”

-0.12’).

4) In order

to synthesize

a protected

1,4,5-IP3,

trio1 such as 2.

(+)-1,2:4,5-bis-cyclohexylidene

of enantiomericallv the requisite 1).

pure

enantiomeric

1,2:5,6di-O-cyclohexylidene (Scheme

the two

Phosphorylation

phenyl

to yield (+)-w-inositol q

trio1 20 from

from

of the esters

to prepare

gave a 90% yield of the bisphosphate

We found that steps

using MPLC (lichroprep

= 16.0’) in near quantitative

for phosphorylating

of racemic

published

obtained

of the

(la],

No

-12L and

1,4_bisphosphate (2) has been 12 (s) by Shvets. The resolution

followed

Basic hydrolysis

chloride

I,4-bisphosphate

resolution of an inositol intermediate. most

fi

= -1.39.

alcohol

Attempts -16D and -16L in 60% overall yield. of mono and bisesters, and the diastereomeric

bisphosphate

1,4,5-trisphosphates.

an efficient also

esters 5.

of the cyclohexylidene

= 0.120, and (-1-2m o-inositol

necessary

D-winositol myo-inositol

by flash chromatography.

of each enantiomer

([al,

3)

of the J-alcohol

gave a mixture

separable

Hydrogeneolysis

$-phosphate,

([aID with

to give the resolved dial. However, -llD could be fully deprotected convenient to treat diol g with two equivalents of S-(-)-camphanic acid

Silica 60) to yield the two camphanate

enantiomeric

(Scheme

salt,

Starting

ester

it was more

were not readily

or 1H NMR.

(+I-myo-inositol

an orthoacetate

to form diastereomeric

the mono-camphanate

Hydrogepolysis of 14D resulted in -14D. cleavage of the ketals, to yield (-l-w-

cyclohexylammonium

1,2:4,5di-O-cyclohexylidene

forming In theory,

that

as its by HPLC

Jl- and L+npy@xbsitoJ l,C-biqhosphates of diol 8 involved

phosphate

by concomitant

was isolated

were

the enantiomeric

Benzvlation

it was Gigg was

mvo-inositol involved a

20 that 1,4,5-trio1

myo-inositol of 1 following

could be

lo (I),

the

Garegg’s

myo-Inositol polyphosphates

SCHEME 3

OH

OH

2D

--c a

Scheme

16D or L

R=

17D or L

R=H

16D or L

R=

Ester

1.4~IP2

Ib 1

PO(OPh),

-AC

3

(4 S-)-camph~ic acid ~l~ide, CH2CL2, (Et13N,DMAP, 25’~; 03) KOH, EtoH, (mO)2PCQ CH2CL2, (Et&N, DMAP, 25Oc;(d) Pt02, EtOH, 1 atm, 25Oc.

250c;fc)

5684

J. P.

VACCA

ef

al,

SCHEME 4

OBtl

OBn Rd.

13.14

BllO

BnO

OP +f

21

P -

P(O)(NHPhX

b

22

R,=H

R*=Bn

30%

23

R&n

R,-H

12%

24

R,=&=Bn

25D

9%

OH

c

260

R-Eater

27D

R-H

25

P-PO$n,

1

1.4.5~IP,

Scheme 4

(a) PhCH2Br,

Bu,+NHSO$, CH2CL2,

CH2CL2,

(Et)3N,

DME/H20

(I:& 25’c,

25’~.

3 h then

DMAP,

25’~;

(c) Acetyl

5% NaOH, chloride,

2 h; (e) NaH, ((BnO)2P(0))20,

AcOH/H20,

25Oc, 18 h.

reflux,

6 h; (b) S(-kamphanic

MeOH/CH2CL2

(I:5 v/v),

25’c,

acid

chloride,

3 h; (d) LiOH,

DME, 0’~; (f) H2, 50 psig, 10% Pd/C,

95% EtOH,

myo-Inositol polyphosphates

procedurelob

provided

the

the 3-benzyl+hydroxyl three

compounds

were

readily

a chemoselective

alkylation

tion

hydroxyl

of the free

3-hydroxyl-4-benzyl

and bis-benzyl

derivative

5685

22 in 39% yield

along

with

formation

of

derivatives

All -23 and -24 in 19% and 9% yields, respectively. by chromatography. Recently Fraser-Reid” has developed

separated

of diol 5 which gives exclusively

the f-hydroxyl

compound

2.

Esterifica-

of -22 with S(-)-camphanic acid chloride yielded a mixture of diastereomers which were chromatographically separated to give -25D and -25L. The diasteroeomeric purity of each compound exceeded 98% as determined both by HPLC and ‘H NMR. All three methyl groups

in the ester

hydrolysis ester

were singlets,

of the trans

group

afforded

With

ketal

phosphate_

the

to

be

chloride,

of bisphosphates

groups

readily

which

to phosphorylate

use of this reagent

nonequivalent

in the diastereomers.

of z

yield.

to facilitate

was successfully

the vicinal

and 4,5-cyclized

in high

removed

inositol

phosphates

It was

in the synthesis

derivatives

due to the lability

after

tedious

these

poor

ion exchange results

by Bartlett.” alkoxide

salt

with

chromatography

to give a dibenzyl-phosphate

alcohols.

Trio1 27D was treated

then

was increased

obtained TBPP.

the

TBPP

realized

the

protected

by simply at

Hydrogenation

L-(-)-myo_inositol Since for

($-mminositol

ambient

the

of cyclized

of 28D rapidly

1,4,5-trisphosphate

our preliminary

the benzyl

1,3,4-t&phosphate for the

preparation

vicinal

reporting

dials

(Scheme

using

The question

vicinally

disposed

in THF (60°C, yield.

at elevated

30 min)

The yield

temperatures

trisphosphate yields

of and

28D was

(>80%) of phosphate

and mild

acid hydrolysis

2oT21 (g).

of

cleaved

In a similar

manner

using trio1 a.

the synthesis either

using similar

of any trio1 from

in high yield.

an

at O°C in DMF in the presence

groups

5) Manipulation

of myo_inositol

with

hydride

1,4,5-trisphosphate

(&) was synthesized

communication

phosphorylating

reported

in high yield

conditions,

salt of 27D was formed removed

was recently

reacted

Even higher

products.

We experienced

problem

salt

these

group. diols is

with trio1 -20 in 44% yield fed to low yield of 1,4,5-IP3

28D in low (25-40%)

tri-alkoxide Under

temperature.

protecting

with vicinal

to phosphorylating

of potassium

trisphosphate

group to give D-(+)-myo_inositol

groups on the various hydroxyls et al allows

equivalents

We found that

and by-products.

deprotected

would be applicable

preforming

when the tri-sodium

the cyclohexylidene

methods

with three

in 65% yield with no evidence

28D were

isomers

which was easily

or not this method

by TBPP to yield

adding

reacted groups

trio1 -27. An answer to the phosphorylation that tetrabenzylpyrophosphate (TBPP)”

was whether

phosphate

myo_inositols

of the blocking

to separate

the

products.

of 4-IP and 1,4-IP2,

of the phosphate

He showed

remaining

followed

Removal

that

final

as well as trio1 27D gave a mixture

which has also found modest success in phosphorylating 16,17 dianilidophosphoryl chloride. Ozaki found that this reagent 2. l4

important

of the

8 and 2 in low yields I2

A reagent

trisphosphate

also

purification

employed

diols of ketals

with more functionalized

to give hexaanilido

Selective

gave diols -26D and -26L. Basic hydrolysis of the pure triols -27D and -27L. 1,4,5-trio1 in hand, it was critical to develop a procedure for

hydroxyl

groups

Diphenylphosphoryl

were

of each diastereomer

route

three

protecting shown

two of which

enantiomerically

an efficient

phosphorylating

has been

substituent

of 4 other

TBPd5122

groups

or phosphite

have disclosed

triesters.14’23-26

of the ketal,

benzyl

and alkyl protecting

methodology

to that

described

any of the

three

biscyclohexylidene

by Gigg13 ketals.

In

J. P. VACCA et al.

5686

SCHEME 5 OBn Ref

(f)B

13

-

OBn

OBn

32

29

R, =R,=H

30

R, =All Rp=H

31

R, =All R2=Bn

J

a b

I

OBn

OH e

OBn

OH

4

33

1,3,4-IP3

Scheme 5

(a) AllylBr, (d) NaH,

NaOH,

((BnO),P(O)),)o,

F’hH, 80’~; DMF,

0’~;

(b) NaH, k)

PhCH2Br,

DMF;

H2, 50 psig, 10% W/C,

(c) 10%

W/C,

95% EtOH,

P-TsOH, 25’~,

3 h.

M&H,

H20;

5688

J. P. VACCA et al.

the present

case,

2,5,6-tri-O-cycle-hexylidene-myo-inositol

hexylidene-pinositol removal

(!I.

Exhaustive

of the trans ketal, followed

,6-di-O-benzyl-myo-inositol The equatorial

(2)

alcohol

diol

1,3,4-tri-O-ally-I-5,6-di-0-benzyl-myo-inositol axial alkylation

is well precedented

gave the fully protected using palladium

was selectively

alkylated

(30) in 88% yield. in the myo-inositol

myo_inositol

on carbon,

l,Z:S,fi-di-O-cycl& inositol.

with

ally1 bromide

This selectivity

literature.

27

trisodium

salt

Removal of the ally1 groups was accomplished acid in methanol 28 to give 2,5,6-tri-O-benzyl-pinositol

trio1 32 was formed in the 33 was obtained in 70% yield.

myo_inositol-l,3,4-trisph~phate2y

(Scheme

for

2,4,5-trisphosphate

synthesis

myo-inositol improve remove gave

sequence,

in the

racemic

were

(x).13

removed

(2) isolated

6)

to prepare

ketal,

In our previous

As in the was formed

by catalytic

as its pentasodium

Although

both

should be possible

the

deprotection

led

to

racemic

communication,

was derived

manipulation

sequence.

the trans

alkoxide

of

33. When in DMF at -0
of TBPP

Subsequent

similar 37 from

This fully

8 the trio1 -37 needed 1,2:4,5-di-O-cyclohexylidene-

from

to that

described

above.

In an effort

3,4,-di-O-cyclohexylidene-myo_inositol

protected

inositol

was treated

with

to

24, a mild

acid to

and then diallylated

to give -34. Acid hydrolysis of the cis-ketal a sample prepared from compound & Selective

diol -35 which was identical with of the equatorial alcohol provided 36 and subsequent

*inositol trisodium

group

we elected

1,4,5-IP3

selectively

benzylation

2

of myo-inositol

@I by protecting

this

byproduct

presence

alkoxide

(2).

&mminoaitoI-2,4,5_trisphosphate the

over

of the axial alcohol

(2) in 93% yield.

p-toluenesulfonic

of the

tris-dibenzylphosphate

to give

for equatorial

Benzylation

In contrast to the 1,4,5-IP3 phosphorylation, treatment of the tripotassium -32 (65%). trio1 12. with TBPP in THF at room temperature gave poor yields of tris-dibenzylphosphate the

Selective

and removal of the cis ketai provided 1,4-di-0-aByl-5 by Gigg 13 et al through the isopropylidene ketals.

prepared

racemic

from

of 8 gave the fully protected

by benzylation

previously

of this

(32) was prepared

allylation

1,3,4-IP3

case,

good

phosphorylation

at
diallylation yields

of TBPP (64%).

in 75% to give racemic

gave 1,3,4-tri-O-benzylwere

obtained

The nine benzyl

if the

groups

of

form,

it

pinositol-2,4,5-trisphosphate

salt.

1,3,4-IP3

and 2,4,5-IP3

to use the resolved

trisphosphates

were

dials -12D or 12L to prepare

either

obtained compound

in racemic

in enantiomerically

pure form. Conclusion

We have

described

of our investigations, for efficiently preparation common

methodology a new procedure

phosphorylating of myo-inositol

orthoacetal

vicinal

for preparing for resolving dials

a number inositol

was developed.

of inositol derivative

phosphates. was found.

This method

In the course Also, a method

has also found

use in the

1,3,4,5_tetrakisphosphate and other inositol phosphates starting Yb intermediate, the details of which will be reported elsewhere.

from

a

myo-Inositol polyphosphates

section

Experimental

Ail reactions the sodium ketyl hydride.

were carried

out under a nitrogen

of benzophenone.

Triethylamine

N,Ndimethylformamide

to the fraction Melting

boiling points

between

60°C-80°C

unless

shifts

are reported

were

obtained with

precoated

in a solution

AlJ combined before

organic

refers

polarimeter.

silica

plates.

extracts

were

Visualization EtOH/5% dried

on a Buchi rotary

to flash

Thin-layer

evaporator

chromatography.

Mass spectra

were

over

2o

recorded

(+)-1,2:4,5_Dicyclohexylide~~~tol(8>, 4-dicyclohexylidene-pinositol hexeneloa

(500 ml) was heated aqueous

was dissolved induce

(CDC13) 1.39-L83

to afford

Anal. Calcd.

The mother

liquors

500 (Porasil

cartridge,

were

for

evaporated

iodine

sulphate

out on E.

or a by dipping on a hot plate.

or sodium

sulphate

of 50°C or below. on a Perkin

>inositol

ChromaElmer

241

was cooled

5% aqueous

(1 g, 5.2 mmol)

to room temperature,

NaHC03

was dried

to which

(7) and @-1,2:3,

(33 g, 180 mmol), l-ethoxycydo-

(500 ml),

and evaporated

was added

petroleum

was filtered

in dry DMF diluted

with

H20 (500 ml) and to an oil. ether

The oil

(40-69’C)

and recrystallized

8, (15.5 g, 25%) mp 171-173°C (lit”

to

from

174’C); ‘H NMR

(m, IH), 4.07 (t, IH, J = 5.2 Hz), 4.47 (t, IH, J = 4.8 Hz); C18H2*06:

C, 63.51, H, 8.29.

to an oil and subjected

Found:

C, 63.48, H, 8.32.

to chromatography

to yield the remaining

on a Waters

two bis-ketals.

7 (21.8 g, 35%) mp 134-135’C from

acetone/petroleum

ether

‘H NMR (CDC13) 1.30-1.77 (m, 20H), 2.59 (d, lH, J = 4.9 Hz), 2.71 (s, IH), 3.39 (t, IH,

J = 8.9 Hz), 3.85 (q, IH, J = 4.6 Hz), 3.94 (t, IH, 3 = 8.1 Hz), 4.03-4.07 6.4 Hz), 4.46-4.49 Found:

x 3Ocm)

by charring

recorded

acid monohydrate

The solid obtained

30% EtOAc/CH2C12)

(+)-1,2:5,6_Dicyd~~li~~i~toi, i33’C);

were

of myo-inositol

phase

of acetone

(m, IH), 3.98-4.04

MS (EI), m/e 340 (M+).

(lit”

with

compound

HPLC

(3.9mm

(m, 20H), 2.58 (d, IH, = 9 Hz), 2.92 (s, IH), 3.30 (t, lH, J = 9.5 Hz), 3.81 (t, IH,

J = 9.9 Hz), 3.86-3.89

Prep

amount

indicated.

a VG70-250.

The organic

of the 1,2:4,5-bis-ketal. ether

magnesium

A mixture

sequentially

NaCL (500 ml).

in the minimum

acetone/petroIeum

anhydrous

The solution

at 100°C for 2 h.

crystallization

acid followed

rotations

Chemical

(TLC) was carried

was done with UV light,

and ptoluenesulphonic

CH2C12 (750 ml) and washed saturated

chromatography

(+)-1,2:5,6dicyclohexylid~

(9)~“~

(60 g, 430 mmol)

unless otherwise

with bath temperature

with

refers

‘H NMR and 3’P

x 3Ocm) or Bondapak-NH2

sulfuric

Optical

Petrol

NT-360 spectrometer.

standard

(3.9mm

from

calcium

indicated.

AM360 or a Nicolet

Porasil

from

sieves.

tubes and are uncorrected.

to Me4Si as internal

of 5% vanillin/90%

evaporation

togrpahy

relative

HPLC using

UV (A254) or RI detection.

Merck 60F-254 plates

as values

on a Waters

was distilled

and distilling

over 4A molecular

otherwise

in open capillary

were determined

Tetrahydrofuran

atmosphere.

was dried by refluxing

(DMF) was dried by storing

NMR were recorded on a Varian XL-300, Brucker

columns

5689

(m, IH); MS (EI), m/e

C, 63.43, H, 8.29.

340 (M+).

Anal. Calcd.

(m, IH), 4.32 (t, lH, J =

for C,8H2806:

C, 63.51, H, 8.29.

5690

J. P. VACCA et .al.

(+)-1,2:3,4_Dicydohexylide~~inositol, (lit”

157’C); ‘H NMR (CDCl3):

2 (13 g, 21%) mp 157-158’C from

1.30-1.78 (m, 20H), 2.88 (s, ZH), 3.68-3.79

9.5 HZ), 4.19 (t, IH, J= 5.5 Hz), 4.62-4.64 C, 63.51, H, 8.29.

Found:

C, 63.39,

(m, 1H); MS (EI), m/e 340 (M+).

acetone/petroleum

Anal. Calcd.

(+~~nyl-~2:4,Micyd~hexyli~~~i~tol

o@h To a solution

of g (12 g, 35.4 mmol) in toluene

by benzyl

The reaction

at room temperature

reaction

mixture

organic gel,

at reflux

was washed

for 6 h followed

with

H20

phase was dried and evaporated

25% EtOAc/petroleum

running

3,6-b&-benzylated

NMR (CDC13):

ether)

for C181i2806:

H, 8.27.

(200 mL) was added NaH (80% in oil, 1.17 g, 39 mmol) followed was heated

ether

(m, 3H), 3.97 (t, IH, J =

by stirring

(2 x 150 mL) and saturated

to an oil.

using

and 6-benzylated

aqueous

This was subjected

a slow flow

rate

compounds

which

yielding

bromide

(4.6 mL, 39 mmol). overnight.

The

NaCl (150 mL).

The

to flash chromatography enabled

separation

(silica

of the

faster

pure lo (9 g, 60%) as a glass.

‘H

6 1.30-1.70 (m, 20H), 2.64 (d, IH, J = 2.8 Hz), 3.26 (t, IH, J = 9.9 Hz), 3.77 (dd, IH,

J = 4.2, 10.1 Hz), 3.88 (dt, IH, J = 2.8, 8.6 Hz), 3.93 (t, IH, J = 5.6 Hz), 4.03 (t, IH, J = 9.7 Hz), 4.35 (t, IH, J (uPorasi1

q

4.5 Hz), 4.82 (d, lH, J = 12.5 Hz), 4.90 (d, lH, J = 12.5 Hz), 7.25-7.43

25% EtOAc/hexane,

2 mL/min).

MS (CI) m/e

calcd.

for C25H3406(M+):

(m, 5H); HPLC 430.2355;

Found

430.2315. Resolutionof (+&“Zyl-l,2:4,Micydohoxylide~~i~itol: in CH2C12 (250 mL) was added (7.7 mL, 55 mmol) stirred

at room

and saturated

temperature

of the diastereomeric yielded

and (-)-camphanic

NaCl solution

To a solution

4-dimethylaminopyridine acid chloride

camphanate

The organic

esters.

the more polar diastereomer

(4.9 g, 22.3 mmol)

The reaction

for 24 h. (100 mL).

Three

of g (8 g, 18.6 mmol)

(DMAP) (250 mg, 2 mmol), mixture

was washed

with

phase was dried and evaporated recrystallizations

!Q (4 g, 36%).

mp 231-233’C;

triethylamine

and the reaction

from petroleum

mixture

H20

was

(2 x 100 mL)

to give a mixture ether

EtOAc

(1:2)

‘H NMR (CDC13) 6 1.00 (s, 3H),

1.05 (s, 3H), 1.11 (s, 3H), 1.26 - 1.76 (m, 2OH), 1.80 - 1.84 (m, IH), 1.86 - 1.98 (m, IH), 2.04 - 2.14 (m, IH), 2.42 - 2.50 (m, IH), 3.38 (t, IH, J = 10.2 Hz), 3.77 (dd, IH, J = 4.1, 10.2 Hz), 4.03 (t, IH, 4.6 Hz), 4.15 (t, lH, J = 9.7 Hz), 4.33 (t, lH, 3 = 4.4 Hz), 4.80 (d, IH, J = 12.5 Hz), 4.90 (d, IH, J = 2.5 Hz), 5.38 (dd, IH, J = 7.1, 11.2 Hz), 7.25 - 7.43 (m, SH); MS (FAB) m/e 25% EtOAc/hexane,

2 mL/min);

C, 68.83, H, 7.59.

Found:

MPLC of the mother 5 mL/min]

yielded

[a]:

-30’

+ 0.6’ (C = 0.45, CHC13).

610 (M+); HPLC (uPorasi1,

Anal

. Calcd.

for C35H460q:

C, 69.09, H, 7.63. liquors

on silica

gel [Lichroprep

69 (Merck),

25% EtOAc/petroleum

ether,

the less polar camphanate

‘H NMR (CDC13) 6 -1lL (3.5 g, 31%). mp 228-23O’C; 1.00 (s, 3H), 1.04 (s, 3H), 1.11 (s, 3H), 1.26 - 1.76 (m, 2OH), 1.79 -1.84 (m, lH), 1.86 - 1.97 (m, IH), 2.04 -2.12 (m, IH), 2.44 - 2.53 (m, IH), 3.38 (t, IH, J = 9.4 Hz), 3.77 (dd, IH, J = 4.1, 10.1 HZ), 4.09 - 4.18 (m, 2H), 4.35 (t, IH, J = 4.4 Hz), 4.81 (d, IH, J = 12.5 Hz), 4.89 (d, IH, J = 12.5 HZ), 5.36 (dd, IH, J = 7.0, 11.2 HZ),

7.43 (m, 5H); MS (FAB) m/e

(M +

(u Porasil,

25%

5691

myo-Inositol polyphosphates C_)_3-Benzyl-l,2~,fi~~~li~~~i~tol (250 mL) was added stirred

the more

organic

(75 mL).

extracts

and evaporated

polar camphanate

The solvent

overnight.

and Et20

(II):

The aqueous

were washed

ester

was evaporated phase

(i&)

and the

was further

with saturated

of KOH (1.4 g, 25 mmol) in EtOH

To a solution

(l.5 g, 2.45 mmol)

residue

extracted

NaCl solution

partitioned

with Et20

(75 mL).

and the reaction

between

water

was

(50 mL)

(75 mL) and the combined

The ethereal

solution

was dried

to afford

‘H NMR (CDC13) 6 1.30 - 1.80 (m, 20H), -12D as a foam (1.05 g, 98%). 2.42 (s, IH), 3.26 (t, IH, 9.9 Hz), 3.77 (dd, IH, J = 4.2, 10.1 Hz), 3.86 - 3.95 (m, 2H), 4.03 (t, IH, J = 9.7 Hz), 4.35 (t, IH, 3 = 4.5 Hz), 4.82 (d, IH, J = 12.5 Hz), 4.90 (d, IH, 12.5 Hz), 7.26 - 7.44 (m, 5H); HPLC (u Porasil,

25% EtOAc/hexane,

Calcd.

(M-H)+ 429.2277;

for c25~3306

2 mL/min); Found:

429.2255.

(+)-~Benzvl-l,2~,5i~~~e~~e~~i~itol less polar camphanate

ester

[aID-26.1 ‘+_ 0.1’ (C = 1.1, CHC13), MS (CI) m/e

(ll): llL., yielded

(m, 2OH), 2.42 (s, IH), 3.26 (TIH,

Repetition

12L as a foam

3 = 9fiz),

of the above procedure,

(90% yield).

using the

1H NMR (CDC13) 6 1.30 - 1.80

3.77 (dd, IH, J = 4.2, 10.1 Hz), 3.86 - 3.94 (m, 2H),

4.03 (t, IH, J = 9.7 Hz), 4.35 (t, IH, J = 4.5 Hz, 4.82 (d, IH, J = 12.5 Hz), 4.90 (d, IH, J = 12.9 Hz), 7.26 - 7.44 (m, 5H). CHC13); MS (CI) m/e

HPLC (uPorasi1, Calcd.

25% EtOAc/hexane,

for C25H3306

of the resolved

mmol),

triethylamine

solution

stirred

mL), followed solutions

alcohol

temperature

by extraction

were

washed

was chromatographed

with [silica

an oil (1.15 g. 75%).

and diphenyl

overnight.

of the aqueous saturated gel,

chlorophosphate

The reaction

phase

NaCl solution

(75 mL) dried ether

= 1.1,

(II):

TO a solu-

DMAP (25 mg, 0.25

(0.8 mL, 3.5 mmol),

mixture

was washed

with CH2C12 (2 x 50 mL).

EtOAc/petroleum

0.1’ (C

25.9’~

429.2235.

2 (1 g, 2.3 mmol) in CH2C12 (50 mL) was added

(I mL, 6.9 mmol)

at room

Found:

4-(6bdiphenylphosphate

(-)-l-(3)-Benzyl-2,3:5,6-(l,2:4,5)-dicycIohexylidene-~inositol tion

[al,

2 mL/min);

(M - H)+ 429.2277;

the title

and the H20 (50

The combined

and evaporated

(1:2)] to yield

with

organic

to an oil. compound

This

E,

as

‘H NMR CDC13 6 1.24 - 1.82 (m, 20H), 3.40 (t, IH, J = 10.1 Hz), 3.75 (dd, IH,

J = 4.1 Hz, 10.2 Hz), 4.05 - 4.13 (m, 2H), 4.33 (t, IH, J = 4.4 Hz), 4.76 - 4.81 (m, 2H), 4.89 (d, IH, J = 12 Hz), 7.16 - 7.42 (m, 15H); HPLC (u Porasil,

25% EtOAc/hexane,

2 mL/min);

[al,

-24.9’

(C

q

0.69, CHC13). (+)-L(3)-kuyl-2,3:5,6-(l,2:4,5)-dicyclohexylidene-mminositol

tion of the above

procedure

using resolved

alcohol

4_(6kliphenylphosphate

E afforded

the title

compound

(II):

(s)

Repeti(75% yield)

‘H NMR (CDC13) 6 1.24 - 1.84 (m, 20H), 3.40 (t, IH, J = 10.1 Hz), 3.75 (dd, IH, J = 4.1,

as an oil.

10.2 Hz), 4.05 - 4.13 (m, 2H), 4.33 (t, IH, J = 4.4 Hz), 4.75 - 4.91 (m, 3H), 7.11 - 7.42 (m, 15H); HPLC (uPorasi1,

25% EtOAc/hexane,

2 mL/min);

(-)_l-Iknzyl-2,3:5,~i~~e~~den~~~itol

[a],.

+ 25.0’

+ 0.2’ (C = 0.73, CHC13).

4-dibenzylphosphate

(II):

To a solution

of the di-

phenylphosphate followed

(13D) (1 g, 1.5 mmol) in THF (25 mL) was added NaH (80% in oil, 99 mg, 3.3 mmol) by benzyl alcohol (0.35 mL, 3.3 mmol). The reaction was stirred at room temperature for

3 h before

being

quenched

with

10% NH4CI solution

(2 mL).

The solvent

was evaporated

and the

5692

J. P. VACCA et al.

residue

partioned

extracted NaCl

with

between

(20

mL) and CH2C12 (20 mL).

CH2C12 (2 x 20 mL).

soluticm

(40 mL),

EtOAc/petroleum the desired

water

ether

dried

The combined

and evaporated

(1:2)] to yield first

dibenzylphosphate

ester

organic

solutions

to an oil.

the monophenyl

(14D) (500 mg,

The aqueous were

This was monobenzyl

50%) as an oil.

phase

washed

was further

with saturated

chromatographed ester

[sifica

gel,

(250 mg, 25%) and then

‘H NMR (CDCl3)

6 1.24 - 1.80

(m, 20H), 3.39 (t, IH, J = 10.1 Hz), 3.75 (dd, IH, J = 4.1, 10.2 Hz), 4.05 - 4.13 (m, 2H), 4.33 (t, IH, J = 4.4 Hz), 4.65 - 4.72 (m, IH), 4.80 (d, IH, J = 12.5 Hz), 4.89 (d, IH, J = 12 Hz), 5.10 -5.20 (m, 4H), 7.26 - 7.43 mL/mln), Found:

[al,

(m, 15H); MS (FAB) m/e

C, 67.77,

599 (M -Bn)+;

HPLC

(uPorasi1,

+ O.l” (C = 1, CHC13). Anal. Calcd. for

= 27.0’

25% EtOAc/hexane,

C39H4709P:

2

C, 67.81, H, 6.86;

H, 6.99.

(+)-1-eenzyl-~3:5,6i~~e~~~~~i~itol

klibenzylphosphate (II):

Repetition of the above

procedure using the diphenylphosphate

-13L yielded the title compound -14L (55% yield) as an oil. ‘H NMR (CDC13) 6 1.24 - 1.80 (m, 2OH), 3.39 (t, IH, J = 9.5 Hz), 3.76 (dd, IH, J = 4.1, 10.2 Hz), 4.05

- 4.14 (m, 2H), 4.33 (t, IH, J = 4.4 Hz), 4.64 - 4.72 (m, JH), 4.80 (d, lH, J = 12.5 Hz), 4.89 (d, IH, J = 12.5 Hz), 5.09 - 5.19 (m, 4H), 7.26 - 7.43 (m, 15H), MS (FAB) m/e 5.99 (M - Bn)+; HPLC (u Porasil, 25% EtOAc/hexane, 67.81, H, 6.86; Found:

mmol) in 10% aqueous

to which excess

was added

was removed

was extracted

(-)-biscyclohexylammonium

180 - 200°C

of the fully protected

with dec.;

by filtration

The solvent

cyclohexylamine

cyclohexylamine

to yield mp.

A solution

(100mL).

EtOH

(C = I, CHC13).

EtOH (100 mL) was hydrogenolyzed

The catalyst

50% aqueous

_+ 0.3’

+26.5’

Calcd.

for C39H4709P:

C,

C, 67.80, H, 6.80.

(-)_lnositol Cphosphate (II): overnight.

[al,

2 mL/min);

into

a pad of Celite, and the residue

The solution

Et20

phosphate

over 10% Pd on carbon

through

was evaporated

(5 mL).

inositol

washing

the Celite

dissolved

was stirred

( 2 x 15 mL).

14D (300 mg, 0.4 (150 mg) at 50 psi

for

The aqueous

with

in H20 (20 mL)

4 h, after phase

which

the

was lyophilized

myo_inositol

‘H NMR (D20)

4-phosphate (15D) (100 mg, 50%) as a tan powder. d 1.10 - 1.40 (m, IOH), 1.60 - 2.00 (m, IOH), 3.02 - 3.12

(m, 2H), 3.42 (t, B-1, J = 9.2 Hz), 3.55 (dd, IH, J = 2.9, 10 Hz), 3.63 (dd, IH, 3 = 2.8, 9.7 Hz), 3.71 (t, IH, J = 9.7 Hz), 4.05 - 4.14 (m, 2H); [alD-l.30 75 mM ammonium C6H1306P,

2

x

formate

C6H13N, 1.5 H20:

(+Mnositol 4-phosphate (II): phosphate - 200°C

&

pH 4, 1 mL/min);

yielded

with dec.;

C, 44.53,

Repetition

(C = 5, H20);

above

procedure

myo_inositol

HPLC (u

Bondapak

360 (M + CHA + I).+

H, 8.72 N, 5.77; Found:

of the

(+)-biscyclohexylammonium

‘H NMR (D20):

_+ 0.2’

MS (FAB) m/e

C, 44.53,

Anal. Calcd.

fi

for

H, 8.31, N, 5.65.

using the fully protected

4-phosphate

NH2,

(60% yield).

inositol mp. 180

6 1.10 - 1.40 (m, IOH), 1.60 - 2.00 (m, IOH), 3.02 - 3.12 (m, 2H),

3.42 (t, IH, J = 9.2 Hz), 3.56 (dd, IH, J = 2.9, 10 Hz), 3.63 (dd, IH, J = 2.8, 9.7 Hz), 3.71 (t, IH, J = 9.7 Hz), 4.05 - 4.13 (m, 2H); [aID 1.1’ _+ 0.3O (C = 5, H20). ammonium

formate

2 x C6H13N, H20:

pH 4, 1 mL/min)

HPLC: (u Bondapak

MS (FAB) m/e 360 (M + CHA + I).+

C, 45.37, H, 8.67, N, 5.88; Found:

Anal. Calcd.

C, 45.20, H, 8.47, N, 5.70.

NH2, 75 mM for C6H1306P,

5693

myo-Inositol polyphosphates Resolution of (~~~2~,Mici~~~itd:

To a solution of & (6 g, 17.6 mmol) in CH2C12

(200 mL) was added DMAP (250 mgs, 2 mmol), triethylamine acid chloride (19.1 g, 88.2 mmol), and the reaction washed beige

with water (2 x I50 mL) and saturated solid (II g, 90%).

5% Et20/CH2C12,

applying

was stirred for 24 h.

The reaction

NaCl solution (200 mL), dried

The two diastereomers

5 mL/min)

(35 mL, 250 mmol) and (-)-camphanic

were

separated

3 g of the above

mixture

and evaporated

using MPLC (Lichoprep

solid in 500 mg batches,

was to a

60 (Merck),

to yield the pure

esters. 2,3:5,6-&2:4,5)_Dicyclohexylid 30%). mp. 250 - 330’

ene-mminositol

sub, 338 - 339’C;

1,4_(3,6)_biiamphanate ‘H NMR (CDC13):

(II,

more

Polar isomer,

Ig,

1.00 (s, 3H), 1.02 (s, 3H), 1.06 (s, 3H),

1.09 (s, 3H), 1.13 (s, 3H), 1.37 - 1.74 (m, 20H), 1.77 - 1.80 (m, 2H), 1.91 - 1.99 (m, 2H), 2.07 - 2.13 (m, 2H), 2.43 - 2.53 (m, 2H), 3.52 (t, IH, J

9.4 Hz), 3.50 - 3.55 (m, 2H), 4.62 (t, IH, J = 4.5 Hz), 5.23

q

(dd, IH, J = 4.3, 10.5 Hz), 5.39 (dd, IH, J = 7.0, 11.2 Hz); MS (FAB), m/e 723 (M = Na);+ [al, + 0.5’

(C = 0.2, CHC13); Anal. Calcd.

for C38H52012:

2,3:5,6-&2:4,5)-Dicyclohexylidene-einositol mp. 280 - 320’ sub, 328 - 329’C;

C, 65.13, H, 7.48; Found:

l,4-(3,6Uiicamphanate

‘H NMR (CDC13):

C, 64.90,

(LJ&, less polar isomer,

9.0’

H, 7.45. Ig, 30%).

0.99 (s, 3H), 1.00 (s, 3H), 1.06 (s, 3H), 1.12 (s,

3H), ‘1.13 (s, 3H), 1.14 (s, 3H) 1.38 - 1.80 (m, 20H), 1.92 - 1.97 (m, 2H), 2.04 - 2.16 (m, 2H), 2.46 -2.53 (m, 2H), 3.52 (t, IH, J - 9.4 Hz), 4.14 (t, IH, J = 9.9 Hz), 4.26 (t, IH, J = 4.9 Hz), 4.67 (t, IH, J = 4.6 HZ), 5.20 (dd, IH, J = 4.3, 10.5 HZ), 5.38 (dd, IH, J = 7, 11.2 Hz); MS (FAB), m/e 723 (M = Na);+ [aID -31.0’

+ 0.5’

(C = 0.2, CHC13); Anal.

Calcd.

for C38H520,2:

C, 65.13, H, 7.48; Found:

C,

65.10, H, 7.35. (+)_1,2:4,5-Dicyclohexylident+mminositol ethanol solvent

(200 mL) was was evaporated

aqueous

phase

(II):

To a solution of KOH (I.1 g, 20 mmol) in absolute

added -16D (750 mg, 1.1 mmol) and the mixture was stirred overnight. and the residue partitioned between water (50 mL) and EtOAc (50 mL).

was further

extracted

with

EtOAc

(2 x 50 mL) and the combined

organic

The The

solutions

‘H were dried and evaporated to yield the title compound -17D (315 mgs, 86%) as a white foam. NMR (CDC13) 1.40 - 1.86 (m, 20H), 2.32 - 2.56 (m, 2H), 3.31 (t, IH, J = 10 Hz), 3.81 (t, IH, J = 9.7 Hz), 3.85 - 3.90 (m, IH), 3.98 - 4.04 (m, IH), 4.07 (t, IH, J = 5.3 Hz), 4.47 (t, IH, J = 4.8 Hz); MS (EI), m/e

340 (M+); [a],

15.7’ of. 0.1’ (C = 3.65, CHC13).

(-r_1,2~,~Dicydohexylid~~i~tol

(II):

Repetition

of the above procedure, but starting

with

the less polar diastereomer as a white

foam.

-16L (750 mgs, 1.1 mmol) afforded the title compound -17L (365 mgs, 100%) ‘H NMR (CDC13) 1.40 - 1.86 (m, 20H), 2.32 - 2.56 (m, 2H), 3.31 (t, IH, J = 10

Hz), 3.81 (t, IH, J = 9.7 Hz),

3.85 -3.90 (m, IH), 3.98 - 4.04 (m, lH), 4.07 (t, IH, J = 5.3 Hz), 4.47

(t, JH, J = 4.8 Hz); MS (EI), m/e (+)_2,3:5,6_U,2~,5)i~~~tol

340 (M+); [a],

-16.0’ + 0.2’

(C = 3.15, CHC13).

1,4-(3,6&bis(diphenylphosphate)

(lZ&&

of -17D (2.5 g, 7.35 mmol) in CH2C12 (100 mL) was added (DMAP, (90 mgs, 0.73 mmol),

To a solution triethylamine

5694

J. P.

(6 ml, 44 mmol) and diphenyl The reaction

mixture

with CH2C12 (3 x 50 mL). to afford

organic

to an oil.

18 (5.25 g, 89%).

(4.6 mL, 22 mmol)

and the solution

H20 (100mL), followed

with

The combined

(100 mL), dried and evaporated ether)

chlorophosphate

was washed

VACCA et al.

solutions

by extraction

were washed

This was chromatographed

stirred

overnight.

of the aqueous

with saturated

phase

NaCl solution

(silica gel, 40% EtOAc/petroleum

mp 181 - 182OC.; ‘H NMR (CDC13) 1.30 - 1.76 (m, 20H), 3.49 (t,

IH, J = 10 Hz), 4.12 -4.17 (m, 2H), 4.54 (t, IH, J = 4.5 Hz), 4.79 -4.83 (m, IH), 4.87 - 4.92 (m, IH), 7.10 -7.33 Calcd.

(m, 20H).

MS (FAB), m/e

for C42H46012P2:

806 (M = 2H)+; [a],

C, 62.68, H, 5.76; Found:

5.6’

+ 0.2’

(C = 0.52, CHC13); Anal.

C, 62.44, H, 5.82.

(-)-2,3:5,6-&2:4,5)-Dicyclohexylidene-~inositol

1,4-(3,6&bis(diphenylphosphate) &3L): Repetition of with -17L afforded the title compound -18L. mp 181-182°C; ‘H NMR (CDC13) 1.30 - 1.76 (m, 20H), 3.49 (t, IH, J = 10 Hz), 4.12 - 4.17 (m, 2H), 4.54 (t, IH, J = 4.5 Hz), the above procedure

but starting

4.79 - 4.83 (m, IH), 4.87 - 4.92 (m, IH), 7.10 - 7.33 (m, 2OH). -5.7’ + 0.1’ (C = 0.52, CHC13); Anal. Calcd.

m/e 806 ( M = 2H);+ [al,

MS (FAB),

for C42H46012P2:

C, 62.68, H, 5.76; Found:

C, 62.45,

H. 5.80.

(+~~Inositol

1,4-bisphosphate

(s)r

The fully

suspended

in EtOH

pressure.

When hydrogen

uptake

of Celite

and the

pad was washed

evaporated After

Celite

and the residue

stirring

had stopped

dissolved

at room temperature

x 15 mL) and the aqueous

-194’C with dec.;

for

over

with

inositol

platinum

the catalyst

in water

!S& (500 mgs, oxide

was removed

50% aqueous

ethanol

to a white

myo_inositol

powder.

at

by filtration (100 mL).

cyclohexylamine

through

a pad

The filtrate

was extracted 2D

was

atmospheric

cyclohexylamine

Recrystallization

I,$-bisphosphate,

0.6 mmol)

(250 mgs)

(20 mL) to which was added

4 h the excess

phase lyophilized

(+)-tetracyclohexylammonium

afforded

protected

(250 mL) and hydrogenolysed

from

(300 mgs,

was

(5 mL).

into

Et20

(2

water/acetone

66%).

mp 184

‘H NMR (D20) 1.15 - 1.98 (m, 40H), 3.11 -3.13 (m, 4H), 3.46 (t, IH, J = 10 Hz), 3.66

(dd, IH, J = 3, 10 Hz), 3.82 (t, IH, J = 9.5 Hz), 3.90 (dt, IH, J = 3, 10 Hz), 4.12 (q, IH, J = 3, 8 Hz), 4.22 (t, IH, J = 2.8 Hz); MS (FAB), m/e 339 (M-H);+ [al, Calcd.

0.12’

+ 0.05’ (C = 4, H20).

Anal.

for C6H14012P2,4 x C6H13N: C, 48.9,H, 9.03,N, 7.6. Found: C, 48.63,H, 8.95,N, 7.55.

(-)-FInositol

I,4-bisphosphate

(&):

Repetition

of the above procedure

with !&

afforded

compound

-2L. mp 184 - 194’C with dec.; ‘H NMR (D20) 1.15 1.98 (m, 40H), 3.11 - 3.13 (m, 4H), 3.46 (t, IH, J = 10 Hz), 3.66 (dd, IH, J = 3, 10 Hz), 3.82 (t, IH, J = 9.5 Hz), 3.90 (dt, IH, J = 3, 10 Hz), 4.12 (q,

IH, J = 3, 8 Hz), 4.22 (t, lo, H20); Anal. Calcd.

J = 2.8 HZ); MS (FAB), m/e

for CgHl4012P2,

4 x C6H13N:

339 (M-H);+

[al,

C, 48.9, H, 9.03, N, 7.6.

-0.12’

+ 0.05’

(C = 4,

Found: C, 48.6, H, 8.8,

N, 7.51. (&(3)-O-Benzyl-2,3:4,Mi-O-cyclohexylidene-~inositol hexylide~~inositol Bii-ketal

7 (5.0 g, 15 mmol)

tetrabutylammonium

(g),

(+)-4-0-Benzyl-1,2:5,6-di-O-cycL*

(22) and (+)-1(3),6(4Mi-O-Benzyl-2,3:4,5-di-O-cyclohexylidene-~inositol

hydrogen

was heated sulfate

at reflux

with

benzyl

(5.0 g, 15 mmol) in a mixture

bromide

(2.78 mL,

of CH2C12

(2). 24 mmol)

and

(250 mL) and 596 aq

5696

J. P. VACCA et al.

(LbCamphanate Ester Diol (xl.

In the

same

manner

as described

was dissolved

in 30 mL of CHC13 and 30 mL of MeOH and treated

After

23’ the

1 h at

residue

partitioned

and dried. la],

reaction

was quenched

with

CHZCIZ and H20.

between

Chromatography’

(SiO2, EtOAc:

= 26’ (CHC13 C = 1 mg/mL)

saturated

The organic

hexane,

above,

NaHC03 layer

1:3) provided

w

(1.0 g. 1.64 mmol)

with 0.10 mL of acetyl soln,

chloride.

concentrated,

was separated,

and the

washed

with brine

0.66 g of 26L (76%), mp 145 - 9’C.

‘H NMR (CDC13) LO6 (3H, s), 1.15 (3H, s), 1.3 - 1.8 IllH, m), 1.9

- 2.0 (2H, m) 2.44 - 2.56 OH, m), 2.78 (2H, br s), 3.44 (JH, dd, J = 9, 2 Hz), 3.76 (IH, dd, J = 9, 2 Hz), 3.88 (IH, t, J = 9 Hz), 4.08 OH, dd, J = 5, 6 Hz), 4.58 (lH, t, J = 5 Hz), 4.87 (2H, abq, J = 15, 11 Hz), 5.19 (IH, dd, J = 6, 9 Hz), 7.3 - 7.4 (5H, m); MS (FAB) m/e Anal. Cakd.

for C29H3809

C, 65.64; H, 7.22.

Found:

~D)-4-o-benzyl-2,3-~~cydohexylide~~i~itol in a mixture

of H20

g, 6.75 mmol). rf starting

material

trio1 27D

The analytical 1 mg/mL);

was stirred

= 0.8, rf product

with CH2C12 and washed give

with

mp 135-S’;

(DME) (l/l, 10 mL each)

at 23’C until TLC indicated

= 0.7).

The reaction

H20 (2 x 50 mL).

(0.197 g, 88%). sample

(27D). Ester 26D (0.34 g, 0.64 mmol) was dissolved

and 1,2-dimethoxyethane

The reaction

The compound

was obtained

531 (M+l), 553 (M+Na);

C, 65.36; H, 7.48.

was concentrated,

The organic

layer

was sufficiently

by recrystallization

and treated

complete

from

with LiOH (0.16

reaction

(2h) (EtOAc,

the residue

was diluted

was dried

and concentrated

pure for use in the pet.

ether.

[al,

next

to

reaction.

= +21°, (CHC13, C =

’ H NMR 1.4 -1.5 (2H, m), 1.55 - 1.8 (SH, m), 2.54 (IH, d, J = 6 Hz), 2.7 (lH,

brs), 2.9 (IH, brs), 3.44 (IH, ddd, J = 12, 9, 3 Hz), 3.69 (IH, t, J = 7.5 Hz), 3.84 (IH, ddd, J = 12, 9, 3 Hz), 3.97 (IH, m), 4.07 (IH, dd, J = 6 Hz), 4.45 (IH, dd, J = 8, 6 Hz), 4.87 (2H, abq, J = 10 Hz), 7.3 - 7.5 (5H, m).

Anal.

Calcd.

for

C19H2606

(L~4-o-benzyl-2,~-o-cyclohexylidene-m_Y ester

26D (0.66 g, 1.25 mmol)

mg/mLTH

gave

C, 65.12; H, 7.48. (s):

In the same

0.42 g (95%) of z,

Found: manner

mp 138-9’C;

C, 64.82; as described

[al,

= -22’

H, 7.39. for 270,

(CHC13, C = 1

NMR (CDC13) 1.4 - 1.5 (2H, m), 1.55 - 1.8 (8H, m), 2.56 (IH, d, J = 5 Hz), 2.83 (IH, br

s), 2.95 (IH, br s), 3.44 (IH, ddd, J = 12, 9, 3 Hz).

3.67 (IH, t, J = 7.5 Hz), 3.84 (IH, ddd, J = 12,

9, 3 Hz), 3.95 (IH, m), 4.05 (IH, dd, J = 6 Hz), 4.44 (IH, dd, J = 8, 6 Hz), 4.85 (2H, abq, J = 10 Hz), 7.2 - 7.4 (5H, m); MS (FAB) m/e H, 7.48. Found:

351 (M+l), 373 (M+Na); Anal. Calcd.

(D)_~~nzyl-2,3-0-cyclohexylide~~~itol 0.5 g of KH (25% in oil) was washed followed

for C19H2606

C, 65.12;

C, 65.19; H, 7.88. 1,4,5-(hexabenzyl)trisphosphate(m). free

of oil with hexane

and to this was added

Method A: 25 mL of THF

The reaction mixture was heated to 60°C for 15 of trio1 (27D). to 23OC and then tetrabenzyl pyrophosphate (TBPP) (0.165 g, 3.08 mmol) was added in

by 0.18 g (0.514 mmol)

min, cooled one portion.

After

3 h, TLC (60% EtOAc/hexane,

The reaction

was complete. dibenzylphosphate,

the

gel (25% EtOAc/hexane) = 1 mg/mL);

was filtered

filtrate

rf sm = 0.2, rf product

through

was evaporated

a pad of celite

and chromatographed

to give 0.378 g (65%) of trisphosphate

MS (FAB) m/e

= 0.35) showed

to remove

on siliCAR-CC4

as an oil. [a]25

the reaction

precipitated

sodium

neutral

= -4.2’,

silica

(CHC13, C

1131 (M+H), 1153 (M+Na); ‘H NMR 1.2 -1.9 (IOH, m), 4.17 (IH, dd, J

q

6,

5691

myo-Inositol polyphosphates

6 Hz), 4.27 (IH, t, J = 6.5 Hz), 4.6 - 4.68 (2H, m), 4.75 - 5..25 (16H, m), 7.1 - 7.35 (35H, m); 3’P NMR (121.4 MHz, CDC13, (MeO13P0 as external

reference, IH decoupled)

-2.01 (s), -2.218 (s), -2.405

(s). Method

BZ The trio1 27D (0.197 g, 0.56 mmol)

2.8 mmol)

precipitate

was added. was observed

reaction

mixture

20 min the suspension EtOAc/hexane)

after

approximately

was concentrated

30 min and the reaction

The filtrate

to give the product

g (68%)

under

an atmosphere

ketal

remained.

still

the

for 18 h.

remaining

that

The residue

The reaction

residue

all of the

was dissolved

was added.

The solution

was diluted

[a],

(H20,

= -27’

(H20,

and chromatographed

with

This was shaken were

in 3 mL of acetic

MeOH to precipitate

pH = 9); ‘H NMR (D20,

(s, 7.47 (s); (1H coupled)

MS (FAB) m/e Found :

triturated

and

with CH2C12, and

6-7 eq of 10% NaOH soln

the product

pH = 10 (cyclohexylamine

as a hexa-sodium

added

to insure

salt

basicity)

pH = 9) 3.64 (lH, dd, J = 10, 3 Hz), 3.83 (3H, brm), pH = 9, IH decoupled)

5.64 (d, J = 3.5 Hz), 7.29 (d, 3 = 7 Hz), 7.39 (d, J = 7 Hz);

421 (M+H), 443 (M+Na).

Anal.

Calcd.

for

C6HgO15Na6P3

3H20

l&+,5-trisphosphate6~):

described

above

[al D + 35’ (H20, C6HgO15Na6P3

Trisphosphate -28L (0.4 g, 0.35 mmol) for 28D to give 0.14 g of & isolated as a hexa-sodium

C = I mg/mL,

5H20

pH = IO), MS (FAB) m/e

C, 11.22; H, 2.98.

Found:

(+I-1,3,4-tri-O-allyl-5,6-di-O-benzyl-pinositol (0.216 ml, 2.5 mmol) and powdered for

‘H NMR as well

but the cyclohexylidene

C, 11.89; H, 2.50.

C, 12.28; H, 2.74.

(LI-pinositol manner

in 50

apparatus

acid and 3 mL of water

the residue

of H20 to which

amount

C = 1 mg/mL,

A crude

removed

4.08 (lH, q, 3 = 9 Hz), 4.26 (IH, brd, J = 1.5 Hz); 3’P NMR (121.7 MHz, D20, 5.67 (s), 7.32

(Si02, 70%

on a mini-Parr

and concentrated. groups

to dryness,

in a minimum

tri-hydrate. Lit7 [al,

benzyl

was concentrated

dissolved

= -30’

The

= -4.5’).

28D (0.49 g, 0.434 mmol) was dissolved

was added.

of H2 (50 psig) for 3 hrs, filtered indicated

1 h.

the L-trio1 -27L (0.25 g, 0.714 mmol) gave 0.545 MS (FAB) m/e 1131 (M+l), 1153 (M+Na).

Trisphosphate

and 0.5 g of 10% W/C

spectrum

stirred

of D-trio&

I&5-trisphosphate (ll):

as a mass

within

in CH2C12 and after

+6.5 (C = 2.8 mg/ml CHC13).

of 28L as an oil, [a],

(Dhnminositol

(80%, [aID

was complete was stirred

1,4,5-bexabenzyl)trisphosphate(28J_): Using Method

above for the phosphorylation

mL of 90% EtOH

The residue

was concentrated

28D as an oil.

(L)_4-o-benzyl-2,~~~e~lide~~i~itol A described

A

to O°C and NaH (0.27 g, 50% in oil) was added.

under high-vacuum.

was filtered.

in 25 mL of DMF and TBPP (1.51 g,

was dissolved

The reaction was cooled

48 h.

The residue

H20 and brine,

421 (M+I), 443 (M+Na).

(33)~ A mixture

then dried and concentrated

between

salt

in the

penta-hydrate.

Anal. Calcd.

for

C, 10.87; H, 2.70.

of 2

(0.93 g, 2.1 mmol), ally1 bromide

NaOH (0.12 g, 3 mmol) in benzene

was partitioned

was treated

benzene

to dryness

and H20,

(50 mL) was heated the

organic

to give 0.89 g (88%) of 30.

layer

at reflux

washed

with

‘H NMR (CDCl,)

3.25 - 3.35 (2H, m), 3.39 (IH, t, J = 9.5 Hz), 3.81 (IH, t, J = 9.5 Hz), 3.91 (IH, t, J = 9.5 Hz), 4.1

5698

J. P. VACCA et al.

- 4.25 (4H, m), 4.25 - 4.4 (2H, m), 4.8 - 4.9 (4H, m), 5.1 - 5.4 (6H, m), 5.9 - 6.05 (3H, m), 7.2 -7.4 (IOH, m), MS (FAB) m/e 481 (M+l).

Anal. Calcd.

for C29H3606

C, 72.48; H, 7.55.

Found:

C, 71.74;

H, 7.62. (+)-1,3,~~i~a5,~~i~~~l-~~itol solution After

of @)

bromide

mixture

washed

was

(1.50 mL, 12.5 mmol)

added

with H20 and brine,

(hexane:EtOAc,

to crushed then dried

6:l) provided

ice

g, 12.5 mmol)

temperature

and stirring

and extracted

with

2.22 g (93%) of 31 as a colorless

for C32H4206

p-toluenesulfonic

The reaction

concentrated.

Found:

(33): A mixture

acid monohydrate mixture

through

was partitioned

with brine, dried and concentrated.

layer

Flash chromatography

oil.

‘H NMR (CDCI,)

The was

on Si02

3.2 - 3.3 (2H,

MS (FAB) m/e

571 (M+I).

C, 75.10; H, 7.51.

of 2 (0.5 g, 0.88 mmol), 10% Pd on C (0.5 g) and

(0.5 g) in MeOH @30 mL)-H20

was filtered

The mixture

stirring.

for 18 h.

The organic

to a

9.5 Hz), 4.0 - 4.1 (6H, m), 4.3 - 4.4 (2H, m), 4.8 -4.9

q

C, 75.75; H, 7.42.

(~~~4,~tri_0_benzyl-~i~tol

added

with magnetic

CH2C12

to dryness.

was

was continued

5.1 - 5.45 (6H, m), 5.85 - 6.05 (3H, m), 7.2 - 7.45 (15H, m).

Anal. Calcd.

72 h.

was added

and concentrated

ml, 3.39 (IH, t, J = 9.5 Hz), 3.88 (IH, t, J (6H, ml,

NaH (0.598

(2.0 g, 4.16 mmol) in DMF (50 mL) at ambient

1 h benzyl

reaction

(31):

filter

between

(20 mL) was heated

ccl, washed

with MeOH

CH2C12 and H20.

Flash chromatography

provided

at reflux

and the filtrate

The organic

layer

for was

was washed

0.31 g (78%) of )2, mp 126 -128’C.

‘H NMR (CDC13) 2.36 (IH, d, J = 7 Hz), 2.33 (IH, d, J = 5.5 Hz), 2.45 (IH, d, J = 2 Hz, 3.35 (IH, t, J = 9 Hz), 3.4 - 3.5 (IH, m), 3.6 - 3.65 (IH, m), 3.79 OH, t, J = 9.5 Hz), 3.85 (IH, td, J = 2, 9 Hz), 4.03 (IH, 5, J = 3 Hz), 4.75 - 4.95 (6H, m), 7.3 - 7.4 (15H, m). 71.98; 6.71.

Found:

C, 72.35;

&mna-benzyl-1,3,4-~inositol-t&phosphate

(2):

Compound

lated

(Si02,

EtOAc:

using method

‘H NMR (CDCI,) (20H,

m),

B.

Flash chromatography m).

(45H,

(&1,3,4-~imsitol-trisphosphate

The reaction residue

precipitate

mixture

(4):

was filtered in H20,

The residue

A mixture

.85, 5.42.

through

treated

was dissolved

filter

apparatus

3’P NMR Calcd.

and 10% Pd on C (0.22

for 6 h at ambient

ccl and concentrated

in a minimum salt.

amount

to dryness

for C6H,0015P3

5Na 2H20

of H20 then

‘H NMR (D,O)

(121.4 MHz, D20, 83% H3P04

1232 (M+l).

-1.89 MS (FAB) m/e

with 6 eq of IN NaOH (1.07 mL), filtered,

0.065 g (64%) of 4 as its penta-Na

Anal.

1:l) gave 0.338 g (80%) of 2.

of 21 (0.22 g, 0.18 mmol)

on a Parr

Hz), 3.83 (IH, t, J = 9.7 Hz), 3.97 (2H, m), 4.18 (IH, dt, 2.44, 2.52 Hz).

C,

32 (150 mg, 0.34 mmol) was phosphoryhexane,

3’P NMR (CDC13) -1.31, -1.45,

(15 mL) was hydrogenated

was dissolved

to dryness.

for C27H3006

3.47 (IH, t, J = 9.5 Hz), 4.05 (lH, t, J = 9.5 Hz), 4.2 - 4.35 (2H, ml, 4.6 - 5.0

7.0 - 7.4

g) in 95% EtOH

Anal. Calcd.

H, 6.90.

temperature.

at <30°C. then

treated

The

concentrated with

MeOH to

(360 MHz) 3.55 (IH, t, J = 9

J = 7.62, 9.21, 9.31 Hz), 4.46 (IH, t, J = as external

reference,

C, 12.73; H, 2.49.

Found:

IH decoupled)

3.87,

C, 12.66; H, 2.69.

myo-Inositol polyphosphates O_5,~i-o-allyl-I,rMi-o-benzyl-~~~l

(34).

Step 1.

CHC13 (50 mL) was added to MeOH (50 mL) containing room

temperature.

concentrated

After

to dryness,

was separated,

washed

1 h the

reaction

and the residue with brine,

mixture

partitioned

dried

and then

0-benzyl-1,2-cyclohexylidene-mzinositol,

5699 (2.21 g, 4.24 mmol) in

A solution of E

3 drops was

of acetyl

neutralized

between

chloride

with

aq.

with stirring NaHC03

CH2C12 and H20.

concentrated

mp 114-16’C after

at

solution,

The organic

layer

to give 1.27 g (68%) of (+) 3,4-di-l trituration with pet ether. H NMR

(CDC13) 1.4 - 1.8 (lOH, m); 2.62 (2H, dd, J = 2.2, 6 Hz); 3.30 (IH, t, J = 9 Hz); 3.7 - 3.8 (3H, m); 3.92 (IH, dd, J = 7.5, 5 Hz); 4.31 (IH, t, J = 4.5 Hz); 4.76 (2H, s); 4.84 (2H, abq); 7.25 - 7.4 (IOH, m).

MS (FAB) m/e

441 (M+l); 463 (M+Na).

Anal. calcd.

for C, 70.89;

H, 7.32.

Found:

C, 70.58;

H, 7.47. Step 2.

NaH (0.835 g, 17.4 mmol)

hexylidene-einositol 1.5 h ally1 bromide was left

to stir

was added

to a solution

added

to ice:water

and extracted

EtOAc:hexane

1:3).

After

at room temperature.

(1.5 mL, 17.4 mmol) in DMF (5 mL) was added dropwise.

for 18 h, then

was washed with H20, brine, dried and concentrated (Si02,

of (2) 3,4-di-O-benzyl-l,2-cyclo-

(1.27 g, 2.9 mmol) in DMF (50 mL) with stirring

The reaction

with CH2C12,

to give 1.2 g (79%) of 34 after

mixture

The organic

layer

flash chromatography

‘H NMR (CDC13) 1.3 - 1.8 (lOH, m); 3.21 (IH, t, J = 9.4 Hz); 3.6 - 3.7

(2H, m); 3.86 (lH, t, J = 8.5 Hz); 3.96 (lH, dd, 3 = 5.4, 7.2 Hz); 4.2 - 4.4 (5H, m); 4.7 - 4.85 (4H, m); 5.1 - 5.35 (4H, m); 5.9 - 6.05 (2H, m); 7.25 -7.4 (IOH, m).

MS (FAB) m/e

~t)-3,edi-O-benzyl-5,6di-o-allyl-~i~itol

(1.2 g, 2.3 mmol),

mL) and H20 ice:water

(15 mL) were

to precipitate

(35):

heated

at reflux

0.817 g (81%) of 2,

Compound

z

for 1 h, cooled

mp 88-90°C

to room

(hexane).

521 (M+l).

glacial

temperature,

then

HOAc (15 added

to

‘H NMR (CDC13) 2.5 - 2.55 (2H,

m); 3.28 (lH, t, J = 9.5 Hz); 3.41 (2H, dd, J = 3, 6.5 Hz); 3.67 (lH, t, J = 9.5 Hz); 3.88 (lH, t, J = 9.5 Hz); 4.21 (IH, t, J = 3 Hz); 4.25 - 4.5 (4H, m); 4.72 (2H, abq); 4.85 (2H, abq); 5.15 - 5.35 (4H, m); 5.9 - 6.05 (2H, m); 7.25 - 7.4 (lOH, m). for C26H3206

C, 70.89; H, 7.32.

Found:

C, 70.?2;

(+)_5,6-di-O-allyl-l,3,4-tri4Lbenzyl-~inositol

inositol

(2)

(0.77 g, 1.7 mmol),

4 mmol) in benzene between

benzene

to provide

and water

and the

441 ((M+l); 463 (M+Na).

A mixture

of 5,6di-O-allyl-1,4di-O-benzyl-myo-

NaOH (0.176 g, 4.4 mmol)

at reflux organic

0.80 g (89%) of 36 as a pale

with stirring

layer

yellow

Anal. Calcd.

H, 7.51.

06):

powdered

(50 mL) was heated

MS (FAB) m/e

for 18 h.

was washed

oil.

with

and benzyl

chloride

The mixture brine,

dried

(0.46 mL,

was partitioned

and concentration

‘H NMR (CDC13) 2.43 (lH, s), 3.2 - 3.5 (3H,

m), 3.79 (lH, t, J = 9.5 Hz), 3.90 (lH, t, J = 9.5 Hz), 4.18 (IH, d, J = 2 Hz), 4.3 - 4.85 (lOH, m), 5.1 - 5.35 (4H, m), 5.9 - 6.1, (2H, m), 7.2 - 7.4 (15 H, m). 0_1,3,4-tri-0-benzyl-~inositol

toluenesulfonic h, then

filtered

acid monohydrate through

filter

(27):

A mixture

MS (FAB) m/e

of 36 (0.8 g), 10% Pd on C (0.08 g and p-

(80 mg) in MeOH (200 mL)-H20 ccl, washed

with

531 (M+l), 553 (M+Na).

(40 mL) was heated

MeOH and the filtrate

concentrated.

at reflux

72

The mixture

5700

J. P. VACCA et al.

was partitioned solution,

between

H20

and Et20

and the organic

brine, and dried and concentrated

with Et20-hexane,

mp 102-104°C.

layer

(R-I, td, J = 2.3, 9.5 Hz), 4.26 (lH, t, J = 2.6 Hz), 4.6-5.0 for C27H3006

+ona-O-benzyl-2,4,~~in06itol

upon tritUratiOn

C, 71.98; H, 6.71.

trlsph~ate

03):

(2H, m), 3.84 (IH, t, J = 9.5 Hz), 3.98

(6H, m), 7.3-7.5 Found:

(ISH, m).

MS (FAB) m/e

C, 72.23; H, 6.76.

NaH (0.102 g, 2.1 mmol)

was

added

to a

of -37 (0.16 g, 0.36 mmol) and TBPP (1.15 g, 2.1 mmol) in DMF (10 mL) with stirring at -2O’C. stirring at -2O’C for 4 h the reaction mixture was treated with 3 drops of 6N HCl and

mixture After

concentrated off.

NaHC03

with saturated

‘H NMR (CDC13) 2.33 (IH, d, J = 5.5 Hz), 2.36 (IH, d, J = 7 Hz),

2.47 (IH, s), 2.6 (IH, s), 3.25 (lH, dd, J = 2.7, 9.2 Hz), 3.4-3.5 473 (M+Na); Anal. Calcd.

was washed

to provide 22, 0.33 g (49%), which solidified

at

<30°C.

The filtrate

The residue

was concentrated

was triturated

with

and chromatographed

CH2C12, and the (Flash,

SO2

resulting

EtOAc:

solid

hexane,

filtered

1:l) to give

0.283 g (64%) of -38. ‘H NMR (CDC13) 3.51 (2H, br t, J = 5Hz), 3.90 (IH, t, 3 = 5Hz), 4.55 (lH, t, J = 5Hz), 4.6-5.15 (lYH, m), 5.37 (IH, br d, J = 5Hz), 7.0-7.5 (45H, m). 3’P NMR (CDC13 -1.526, -1.572, -1.761.

MS (FAB) m/e

1231 (M+l).

Anal. Calcd.

for C6yH6y015P3

C, 67.31; H, 5.65.

Found:

C, 67.63; H, 6.17.

(+)_2,4,~~inoJitol_triphoaphate g) in 95% EtOH was hydrogenated reaction

was filtered

dissolved

in distilled

concentrated

through H20,

to dryness.

A mixture

(I):

in a mini-Parr

filter-ccl treated

Trituration

of 38 (0.19 g, 0.15 mmol) app:atus

and concentrated

with with

0.926

and 10% Pd on C (0.19

for 5 h at ambient to dryness

mL of 1N NaOH soln

temperature.

at C30°C. (0.924

The

The residue

mmol),

filtered

MeOH gave 0.032 g (34%) of 2 as its hexasodium

was and salt.

‘H NMR (D20) (360 MHz) 3.48 (lH, dd, J = 2.1, 9.9 Hz), 3.72 (lH, dt, J = 2.5, 2.5, 9.7 Hz), 3.84 (lH, q, J = 7.5, 9.9 Hz), 3.95 (lH, t, J = 9, 10 Hz), 4.23 (IH, q, J = 7.5, 9.5 Hz), 4.45 (lH, dt, 2.5, 7.2 Hz). 2 Hz; 5.23, H, 2.50.

31P NMR (121.4 MHz) (D20) (85% H3P04 P5, 7 Hz.

Found:

MS (FAB) m/3

C, 11.87; H, 2.17.

417 (M+l).

ext.

ref.)

Anal. Calcd.

J = 2,

5.38, p-2, 7 Hz; 5.50, P-4, 7 and for C6HyO15P3.6Na3H20

C, 11.89;

Wo-Inositol polyphosphates

5701

References (1) (2) (3)

Nishizuka,

Y.; Nature

Berridge,

lY@, 308, 693.

M. J.; Irvine,

R. F. Nature(London)

lY84, 312, 315-321.

and references

therein.

Rev. 1986, 3

227-272

The structures

shown in Figure 1 are drawn as their absolute

are

given

the

are depicted

D- prefix in their

cited

regardless

absolute

of optical

configuration

Abdel-Latif,

configuration.

All of the Compounds

All compounds

rotation.

but actually

are racemic

A. A. Pharmacol.

throughout

mixtures

the

unless

paper

otherwise

indicated. (4)

Storey,

(5)

Berridge,

D.

(6)

Batty,

J.;

Shears,

L R.; Nahorski,

A. J.; Laoder, (7)

Crado, Vacca,

Billington, a)

Michell,

R. F. Biochem.

C. P. Biochem.

S. J.; Huff,

J. P.; Huff,

R.

H. NaturefLondon)

lY84, 312,

374.

J. 1985, 232, 2ll-215; Irvine,

R. F.; Letcher,

J., lY84, 223, 237-243.

l%l,

236, 54-60; Irvine,

R.; Kulogowski,

Billington,

D. C.; Baker,

A.; Akunian,

P. J.;

Iverson,

E

Stepanov,

(12)

Krylova,

(13)

Gigg, J.; Cigg, R.; Payne,

Lett.

J. J.; Mawer,

R.;

R. F.; Brown, K. D.; Berridge,

Etat.

lY87, 109, 3478-3479.

deSolms,

4505-4506. L M. J. Chem. Chem.

Sot., R. S

Lindberg,

B. A.; Shvets,

A. E.; Klyashchitskii,

Sx.

1987, 3

R. J. Chem.

Bull. Univ.

T.; Johansson,

(II)

Chem.

J. R. J. Amer.

J. R. Tetrahedron

D. C.; Baker,

Johanissian,

Garegg,

3;

J. 1984, 223, 269-272.

J. P.; deSolms,

1987, 314-316. (10)

S. R.; Irvine,

C.; Ballou, C. E., J. Biol. Chem.

S. J.; Vacca, (9)

C.

J. 19114, 220, 345.

D. J.; Dowries,

M. J. B&hem. (8)

S. B.; Kirk,

M. J. Biochem.

S. Armenie

B. Carbohydr.

V. I.; Evstigneeva,

Sot.,

Commun.

Chem.

Comm.

19117, 1011.

1930, 5, 245-249;

Res.

h)

lY84, 130, 322-326.

R. P. Bioorg.

Khim.

1976, 2,

1627. N. I.; Oleinik,

V. N.; Kobel’kova,

G. F.; Shvets,

V. I. J. Org. Chem.

USSR 1980, 4

59.

(14)

Payne,

S.; Conant,

Ozaki,

S.; Watanabe,

Tetrahedron

Lett.

S.; Conant,

R., J. Chem. Y.;

Ogasawara,

Yu, K., Fraser-Reid, Synthetic

Comin.

198% 5

(16)

Cremlyn,

R.

W.; Dewhurst,

(17)

Krylova, 277. B

B. Tetrahedron

V. N.; Gornaeva,

Krylova,

Perkin

Trans.

T.; Kondo,

Res. 1985, 140, Cls3;

Gigg, J.; Cigg,

R.;

1. lY87, 423-431. Y.;

Shiotani,

N.;

Nishii,

H.; Matsuki,

I

T.

1986, 26, 3156-3160.

(15)

J.

R. Carbchydr.

Sot.

Lett.

1988, 26, 979-982.

Yu, K.; Ko, K.; Fraser-Reid,

8.

465-468.

V. N.; Lyutik,

D. H. J. Chem.

B. B.; Wakeford,

N. P.; Oleinik,

G. F.; Shvets,

A. I.; Gornaeva,

SodC)

V. I. J. Org. Chem.

N. P.; Shvets,

1971,

300-303.

USSR ISSO, If?t

V. I. J. Gen. Chem.

USSR 19111,

183.

(18)

Chouinard,

(19)

Khorana, H. G.; Todd, A. R. J. Chem.

P. M.; Bartlett,

P. A. J. Org. Chem.

(20)

The compound

was identical

to have reported

to that

by: Cerdan,

reported

S.; Hansen,

J. Biol. Chem.

1986, 2,

14676-14680.

J. M. Biochem.

J. 1986, 233, 275-277.

1986, II,

75-78.

Sot. 1953, 2257-2260. in ref.

7a.

Also, the ‘H NMR of 10 was identical

C. A.; Johanson,

Lindon,

R.; Inubushi,

J. C.; Baker,

T.; Williamson,

D. J.; Fat-rant,

J. R.

R. A.; Williams,

5702

J. P. VACCA

biologically 3. P.; deSolms, (22)

Watanabe, Kondo,

Y.; Nakahira,

Tetrahedron (23)

Lett.

1988 263, 11922-11927. Lett.

Y. Tetrahedron

M.; Watanabe,

Lett.

1987, 28, 4179.

Lett.

Y. Chemistry

Lett.

Boehm,

1988, 29, 5305-5308.

1987, 3

Otaki,

4691.

1988, 77-80.

M. F.;

Ley,

Prestwich,

S.;

Ozaki, S.

G. D.

1988, 29, 5217-5220.

C. F.; van der Marel,

Dreef,

S. Tetrahedron

S.; Watanabe,

H.; Bunya,

F. Tetrahedron

M. A.; Bencen, G. H.; Vacca,

1 see Polokoff,

J. R. J. BioL Chem.

H.; Bunya, M.; Ozaki,

H.; Yomooka,

M.; Nakahira,

V.; Stemfeld,

to natural

S. D.; Huff,

S. J.; Young,

Y.; Nakahira,

S; Kohno,

equivalent

al.

C.

A.;

van Boom,

J. H.; Reel.

Trav.

Chim.

Pays-Bas

1987, 106,

161. (24)

Reese,

C. B.; Ward, J. G.; Tetrahedron

(25)

Cooke,

A. M.; Potter,

J.

L.;

1987, 28, 2309.

B. V. L.; Tetrahedron

V. L.; Gigg, R. J. Chem. V. L. J. Chem.

Lett.

Sot. Chem.

Sac., Chem.

(26)

Meek,

Davidson,

(27)

Angyal,

S. 3.; Tate,

kc.(C)

1969, 2367-2371.

Commun

F.;

M. E.

1987, 28, 2305.

1987, 626.

Cooke,

Jr.,

F.

W. J.

Amer.

Sot. 1965, 6949-6955.

(29)

The’H NMR and 31P NMR was identical

to that

Williams,

J. 1987, 244, 591-595.

J. M.; Irvine,

Chem.

Chem.

Fbss,

R. Angew.

Hamblin,

B. B.

R. F. Biochem.

Intl.

!kc.

1988,

Gigg, R.; Warren,

(28)

R.; Scheffold,

M. R.; Potter,

A. M.; Gigg, R.; Potter,

1987, 1525-6.

Hobbs,

J. Chem.

Lett.

Commun.

ll0,

2317-2318.

C. D. J. Chem.

Ed. Engl. 1976, 151 558-559. recently

reported;

Lindon,

J. C.; Baker,

See also ref.

2Oa.

D. J.;