Tetrahedron
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USE OF
AS A PHOSPHORUS
VI A
GROUP I N OLI GONUCLEOTI DE SYNTHESI S
A
PHOSPHI TE TRI ESTER APPROACH
C.
C. E.
J . E.
G. A. va n de r
a nd J . H. va n Boom
Labor at o-
Depar t ment of Chemi s t r y, Uni ver s i t y of Ni j megen, 6525 ED Ni j megen and
P. O. Box 9502, 2300 RA Lei den, The Net her l ands
r i es , St at e Uni ver s i t y of Lei den,
ABSTRACT:
di chl or ophos phi ne has been conver t ed i nt o t he mono- N- mor pho-
l i no der i vat i ve and appl i ed f or t he pr epar at i on of
d- nuc l e os i de - 3' - phos phor -
a mi di t e s . The l a t t e r i nt e r me di a t e s c oul d be us e d i n t he pr e s e nc e of
f or t he f or mat i on of
l i nkages . The MSE pr ot ect i ng gr oup can be r emoved
s el ect i vel y and f as t under mi l d bas i c condi t i ons .
The i nt r oduc t i on of t he phos phi t e
me t hod by
s t i mul a t e d s e ve r a l gr oups
a nd, e s -
t o s t udy i n de t a i l t he a ppl i c a t i on of di f f e r e nt
peci al l y s o, met hyl
s t udi es i ndi cat ed t hat met hyl phos phor odi chl or i di t e ( i . e.
vi t y, be us ed f or t he s ynt hes i s of DNA on s ol i d s uppor t . I n or der t o decr eas e t he r eact i vi t y
c onve r t e d i nt o t he r e l a t i ve l y mor e s t a bl e c hl or o
phos phor a mi di t e s ( e . g.
pr ove d t o be s ui t a bl e f or t he s ynt he s i s of
t owar ds t he s ynt hes i s of DNA. The r es ul t s of t hes e
coul d, des pi t e i t s hi gh r eact i -
a
of a ge nt
me t hyl
a nd a l s o
The l a t t e r a ge nt s , a nd pa r t i -
d- nuc l e os i de
c ul a r l y s o
phor ami di t es ( e. g.
X=N- mor phol i no;
whi ch s er ved as key i nt er medi at es f or t he pr epa-
r a t i on of i mmobi l i z e d DNA f r a gme nt s ( e . g.
R=Me ;
The r e mova l of
mi xt ur e of
t he met hyl P- pr ot ect i ng gr oups f r om
or
can be ef f ect ed by t r eat ment wi t h
a
We now wi s h t o r e por t t ha t : ( i ) t he e a s i l y a c c e s s i bl e a ge nt
l i no phos phor omonochl or i di t e ( I d) has t he s ame phos phi t yl at i ng pr oper t i es as t he met hyl
gue ( i i ) t he ( MSE) gr oup f or pr ot e c t i ng P- O c a n be r e move d s e l e c t i -
vel y under mi l d bas i c condi t i ons .
The f avour abl e pr oper t i es of
as
a
pr ot ect i ng gr oup has been demons t r at ed bef or e i n
t i de
chemi s t r y, we f i r s t l y exami ned t he f eas i bi l i t y of pr epar i ng agent
coul d be obt ai ned as s t abl e cr ys t al l i ne s ol i d by t he f ol l owi ng pr ocedur e. To
mol ar exces s of phos phor us t r i chl or i de i n acet oni t r i l e was added dr opwi s e, dur i ng 20 mi n at
I n or der t o f i nd out i f t hes e pr oper t i es coul d be expl oi t ed i n nucl ei c aci d
I t t ur ned out t hat agent
s evenf ol d
a
a
i n acet on- i t r i l e. Af t er
s mal l vol ume and t he r es i due was di s t i l l ed t o af f or d pur e
0. 05 mm Hg;
t he N- mor phol i no der i vat i ve
at t he r eact i on mi xt ur e was concent r at ed t o af f or d cr ude
t hus obt ai ned by
t yl a t i on of t he
e t
4
h
at
t he s ol ut i on was concent r a-
t ed t o
a
[ yi el d 90%; b. p.
178. 71 whi ch cr ys t al l i zed on s t andi ng. Agent
by t r eat ment
was now conver t ed i nt o
wi t h N- t r i met hyl s i l yl mor phol i ne
as
i n THF. Af t er
1
h
a
gum. Anal ys i s of
s howed t he pr es ence of mai nl y one r es onance ( 6 167. 5) .
d- nuc l e os i de s ( 1 mmol ) , a c c or di ng t o t he me t hod of
i n di chl or omet hane af f or ded, af t er wor k- up and pur i f i cat i on by
wi t h
( 1. 7
col umn- chr omat ogr aphy,
s ol i ds ( yi el ds
t he
phos phor ami di t es
( X=N- mor phol i no;
as homogeneous
bas ed on 2a) . The appl i cabi l i t y of
was demons t r at ed by t he s ynt hes i s
1307
Claesen, C.
