CA1235079A - Process for the preparation of nucleoside alkyl-, aralkyl- and aryl-phosphonites and -phosphonates - Google Patents

Process for the preparation of nucleoside alkyl-, aralkyl- and aryl-phosphonites and -phosphonates

Info

Publication number
CA1235079A
CA1235079A CA000462469A CA462469A CA1235079A CA 1235079 A CA1235079 A CA 1235079A CA 000462469 A CA000462469 A CA 000462469A CA 462469 A CA462469 A CA 462469A CA 1235079 A CA1235079 A CA 1235079A
Authority
CA
Canada
Prior art keywords
general formula
group
compounds
formula
alkyl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired
Application number
CA000462469A
Other languages
French (fr)
Inventor
Joachim Engels
Alfred Jager
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hoechst AG
Original Assignee
Hoechst AG
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hoechst AG filed Critical Hoechst AG
Application granted granted Critical
Publication of CA1235079A publication Critical patent/CA1235079A/en
Expired legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/48—Phosphonous acids [RP(OH)2] including [RHP(=O)(OH)]; Thiophosphonous acids including [RP(SH)2], [RHP(=S)(SH)]; Derivatives thereof
    • C07F9/4883—Amides or esteramides thereof, e.g. RP(NR'2)2 or RP(XR')(NR''2) (X = O, S)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Saccharide Compounds (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)
  • Silicon Polymers (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)

Abstract

Abstract of the disclosure Deoxyribonucleoside phosphonates, thiophosphonates and selenophosphonates are obtained by condensation of a difunctional phosphonylating reagent of the formula R-PXY, in which R is an inert non-cytotoxic organic radical, X
is chlorine or Y and Y is a secondary amino group, with a deoxyribonucleoside of which the 5-hydroxyl group and any exo-amino group present in the base radical are protected, and further condensation with a nucleoside of which the 3-hydroxyl group and any exo-amino group present in the base radical are protected, and then oxidation. The thio-phosphonates and selenophosphonates and the intermediates of the first condensation stage are new.

Description

HOE 83/~ 178 ~35~79 Non-ionic analogs of deoxyribonucleic acids tuna) are important for the investigation o, Deb and EDNA
protein interaction Of particular interest are phase phonic acid esters of deoxyri~onucleotides as a result of their cheInic"l sublet and on the basis of their capably-my to erlt~r Jo cells and their high resistance Jo cell nucleasesn Hitherto, four different strategies have been descr;bcd for the synthesis of methylphospi)onate analogs of nucleotides:
1. Ogilvie et at. MY Newer and OK Ogilvie, Twitter-heron Let. 21~ Page l~149 (1980)) prepared a completely protected uridyl-3',5'-uridine rnethylphosphonate by Michaelis~Arbu~ov rearrangement of the corresponding faceplate intermediates. Tl1is reaction (methyl iodide, 20 hours at 50C3 might not be generally applicable as a result of its drastic conditions, because, for example, methylation of the Purina bases is to be expected.
I Sue et at. (P.S. Miller, J. Yank E. Yank C. Carol, K. German and PUP. Sue, Biochemistry 18, 5134 on ~197~) Prick Nf~tl~ Aged. Skye USA I 1537 (1~81);
US. Miller, N. Drown SUM Pilfered and K.B. McFarland, Jay Blot. Siam. 22~r 9~59 (19~0)~ developed a synthesis strategy wtlIch is analogous to the phosphotr;ester method in ollgonucleotide synthesis. ire, a protected nucleated ~'-O-methylphosphoIl;c acid ~-cyanoethyl ester is used as the most im~crt2nt inter~ediaten Thus method has the , I.

:~35~
! 3 known, advantages and disadvantages of the phosphotriester method, the lo reactivity of the phosphorus TV) compound being mentioned in particular as a disadvantage.
3. Agarwal et at. (CLUE. Agar~lal and F. Rift;na, Null.
Acid Rest 6, 3009 t1979)) used methylphosphonic acid dip chloride as a difunct;onal phosphonylating agent. In the secorld stew the chloride has to be activated by means of tetrazole. The crook product obtained can only be purified by efficient chromatography.
1~1 4. Joy angels and A. Jagger, Anger. Chum. Supply 19~2, 2010 and Do Sweeney, V. Grossbruchhaus and H. Castro, Twitter-heron Lotte I 7 t1983) used methylclichlorophospl1ane as the starting material. The latter authors synthesized the nucleated methylphosphonates on a polymeric support.
The products obtained are yet to be caricatured Whereas the reactivity of the second halogen of methylphosphonic acid d;chlor;de is generally too low and additional activation is necessary the activity in the case of phosphinic acid dichlorides us if anything too on high. Thus handling difficulties arise extremely an-hydrous medium) and, on add ton the symmetrical pros-pharisee acid ester us unavoidably formed.
By contrast, the invention relates to a process for the prepara,;orl of deoxyr;bonucleos;de phosphonates of the general formula I

~35~79 T -I y 1 t I ) o owe G-O
on which T denotes a protecting group for a pruner hydroxyl group, preferably tr;phenylrnethyl I= Try), pencil-diphenyLMethyl or do anisoyl)phenylrnethyl, B denotes a nucleoside base radical on which any ego-amino group present is protected, preferably 1-thyminylr ON ~benzoylcytosinyl), 9-(N-6-benzoyl-adeninyl~ or 9-(N-2-isobutyroylguaninyl)~
G denotes a protecting group for a secondary hydroxyl group, Z denotes oxygen, sulfur or selenium and R denotes alkyd having up to 8 C attorneys cyclohexyl, benzyl,or furl optionally substituted by fluorine, choline bror,1ine~ lower alkyd, lower alkoxy or in-fluoromethyl~ and preferably denotes methyl, ethyl phenol or bouncily especially methyl, he eon a difunctional phosphonylating reagent of the general formula II
R - P y warily X denotes chlorine or Y and Y denotes a group of the formula _ 5 _ ~35~7~
~,~
-of I
R1 and R2 representing identical or different ~lkyl or cycloalkyl radicals having up to 8 C atoms, or phenol ridiculous or R1 and r~2, together with the nitrogen, S representing a saturated or unsaturated heterocyclic ring which can contain further heteroatorns, is reacted with a nucleoside of the general formula JOY

B
T Ox IT

whereon T and B have the rneanincls given above preferably at -80 to +100C, in particular at ~20 to DO the resulting compound of the general forr,lul3 It Jo y (IV) R -- P

is reacted with a compound of tile general formula V

y V ) I; -0 wherein B and G have the rmearl;nçls coven above preferably at -Z0 to ~1nnC~ in particular at room temperature, and the resulting compound of the general formula VI

B lZ35079 I ' . , - o R - P B (VI) by G- O
wherein T, I B and G have the meanings given above, us ox datively converted to compounds of the general formula i 1, preferably at -80 to ~100C, in particular at -20C to room temperature.
The compounds of the general formula I in which Z
denotes sulfur or selenium, and the intermediates of the general formula IV, are new and also form a subject of the ;nvent;on.
In principle, the radical R in the difunctional phosphonyla~ing reagent of the general formula II can be any non-cytotoxic organic radical which is inert towards the compounds of the general formulae II to VI and which does not hinder the reactions.
Examples of possible groups of the general formula -NR1R2 are: d;methylam;no~ d;ethylam;no, diisopropyl-amino, methylethylamino~ methylpropylam;no, methylhexyl-amino methylcyclohexylamino~ methylbenzylam;no, morpholinoO
pyrrolidino, piper;dino, methylanilino, diphenylam;no, I 1midazolo, triazolo, benzotriazolo and tetrazoloO
The starting materials of the general formula II
on which X denotes chlorine can be obtained by reacting the corresponding dichlorophosphane~ preferably methyldi-chlorophosphane, loath a secondary amine of the general formula Eli 7 - ~L23~9 INURE R2 (VII 3 on which R1 and R2 have the meanings given above Correspondingly, compounds of the general formula II in Shea X denotes a group of the formula Y are accessible by further reaction loath the same secondary amine or a different secondary amine of the general formula VII. The compounds of the formula II can be purified by vacuum d;stillat;on.
The reaction of the phosphonylat;ng reagent of the I general formula II with a suitably protected nucleoside of the general forr,lula Ill is carried out in a moderately polar solvent preferably chloroform, with the exclusion of moisture. Tertiary amine, preferably ethyldiisopropyl-amine Hunks Boyce can be used as auxiliary bases for this reaction. Work;ng-up is carried out by aqueous ox-traction and precipitation of the products of the general formula IV with a non-polar solvent such as petroleuln ether or pontoon. The phosphorous acid ester~am;des of the general formula IV obtained on this way precipitate as colorless powders and can be characterized by spectroscopic data such as OH NOR, 3~P-NMR or US and elementary analysis. Furthermore, they can also be converted, by direct oxidation, to the phosphoric acid ester amicles of the general formula VIII

B
T - O

I VOW) Z Jo p _ pi ~35~

T, B, Z, R and Y having the meanings given above, which can then be isolated and characterized.
Remarkab1yr no symmetrical dinucleoside 3',3' phosphonite us formed within the limit of detection.
As shown by 31P-I~MR, the compounds of the general formula I are stable for at least 1 month in poller form, when stored dry and at a maximum of -20C~ This great stability of the phosphorous acid ester-amides is astonisl1-in and emphasizes the value of this method. Its universal applicability in the synthesis of phosphoric acid divesters of nucleosides is shown by the reaction with suitably protected nucleosides:
In this reaction, the protected nucleoside phosphonites of the general formula IV are dissolved in a moderately polar solvent, preferably acetonitrile~ sheller-form or tetrahydrofuran, and mixed with the nucleos,de of the general formula V (protected in the 3l~positionin Suitable protecting groups G in the compounds of the general formula V are azalea groups such as bouncily, acutely, pivaloyl or levulonyl, or sill groups such as t-butyl-d;methyls;lyl. The reaction us catalyzed by an acid, preferably an azalea or amine hydrochloride. Benzotr;azole us particularly suitable. It us remarkable that H~LC
analysts of the product shows no symmetrical Rosemary and only traces of the 3'~3'-isomer;c phosphonate.
he labile ;ntermed;ate, namely the phosphorous acid treater of the general formula VI, is oxidized directly to the phosphonate of the general formula It In addition to the oxid;z;ny agents usually employed for thus _ 9 purpose, such as dinitrogen tetroxide or iodine, peroxide, on particular an hydrous t-butyl hydroperoxide, have proved valuable. The reaction is proofer carried out in a moderately polar solvent, particular preference being S afforded to acetonitrile or chloroform. Particular con siderat;on should be given to the known acid-catalyzed transesterification of the diacylalkylphosphonites {WOW
Hoffman, I Roth and TO Simmons Jo Amer. Chum. Sock 80, 5~37 - It (1958) ) .
The compounds (some of which are already known) are characterized by means of 3~P-NMR and 1H-NMR and also chromatographic comparisons with authentic maternal.
The compounds of the general formula I on which denotes sulfur or selenium are prepared by direct reaction of the compounds of the genera formula VI with elemental sulfur or selenium. Stirring with toe stoichiometric quantity of sulfur or selenium in a polar solvent such as tetrahydro~uran, leads to good yields of the corresponding thiophosphonates or selenophosphonates of the gene at formula I. Characterization is carried out by means of 31P-NMR and 1H-NMR as well as elementary analysis Because of the presence of a center of asymmetry on the nucleoside moiety and the production of another on the phosphorus, the phosphates of the general formula I
exist as mixtures of diastereomers (see Table 6, isomers 1 and Z).
The isomer ratio, ~Jh;ch us close to the statistical ratio of 1:1, is only very slightly influerlced by a variation on the parameters such as the solvent the them-I

portray and the sequence of addition.
The ox am pies which follow describe the invention in greater detail:
Example 1: Starting material H3C-P~N-(CH3)2~
In a 1000 ml three-necked flask fitted with a dropping funnel and a mechanical stirrer, 125 rnl ~1.9 Noel) of dimeti)ylamine are introduced into no ml of an hydrous deathly ether and reacted, over a pyre of 60 minutes with a solution of 60 ml (0.40 molt of methyld-ichlorophosphane on 200 rnl of an hydrous ether, whole cooling with ice. After Sterno for 2 hours at room temperature and for 1 hour at 50C, the precipitate us filtered off under a protective gas and rinsed twice wow 100 ml of ether and the filtrate is concentrated at ablate aye bar. The remaining residue us rapidly distilled over at 0.5 ark. Precision distillation with a Vigreux column ~50 con) at 64-65C/65 mar gives 36.6 9 t66% o-f theory) of a colorless liquid.
Analysis Clue ) < Q~2~
I 31p_NMR thief) = 87 ppm Do = 1.Z3 ppm (do 7Hz, P Clue) S 2.66 ppm id, 7Hz, N(CH3)2) Examples:
The 5'~tritylnucleosides III to Molly) are dissolved in 6 ml of an hydrous chloroform under an inert nitrogen atmosphere and ~l3CP~N(CH3)2~2 (2 Molly) is added. The reaction is complete after 12 hours at room temperature (stirring) or after only 2 hours if catalytic quantities tn.1 Molly) of colliding hydrochloride are added.

.

35i~7~3 The solution is then transferred with 100 ml of ethylene chloride to a 250 ml separating funnel and extracted lice by shaking with 50 Al of saturated sodium chloride solution (containing 0.1 ml of triethylamine).
The organic phase is dried over an hydrous sodium sulfate and concentrated to a foam. This us stirred for 2 hours with 50 ml of pontoon. The residue is filtered off and dissolved in 2 ml of deathly ether and the solution is slowly added drops to 50 ml of thoroughly stirred pen-lane The fine precipitate is filtered off and dried to give an 85-95X yield of the compound of the general formula IV tables 2 and 3).
The compounds can be identified directly by 31p nuclear magnetic resonance spec~roscopy or, after oxidation with t-butyl hydroperoxide, as phosphoric acid ester-amides of the general formula VIII tables 4 and I
In the 31P-NMR spectrum, these substances show up to 3X of hydrolyzed product (nucleos;de methylphos~
fount), but no detectable quantity of symmetrical do nucleos;de 3',3'-phosphoniten Thus demonstrates the superiority of the method compared it former methods, which always yielded about 5-10% of these products. When stored as dry powders at -2~C, no decomposition can be observed within a Month.
The following reagents wore also employed analogously:

~235079 1~3C Pi ( 2H5 H3C -- P f 3 ~-72 C
No 5 3 ( C 6 H 5 ) 2 I Of "
Example 3:
The S'~tritylnucleoside III (1.0n mrnol) and 1.71 ml (10 Molly) of NUN ethyldiisopropy~amine are introduced unto 6 rnl of THY, and 2.00 Molly of phosphonyla-to agent II are then slickly added drops. After stirring at room temperature overnight, the reaction solution is added clropw-ise to suckled water ~50 ml, saturated with Nail). after extraction with twice Z0 ml of ethylene chloride, the organic phase is dried with sodium sulfate and the solvent is removed in vacua Further purification is carried out by precipitation as above tables 2 and I
Example 4-I BenzoylthyMidine ~0.20 Molly) and buoyancy-triazole ~0.80 rnmol) are dried in a round-bottomed flask and then dissolved in 1.0 ml of dry acetonitrile. The reaction is complete within one minute, a very air labile and acid-labile phosphonite I being formed; this is converted directly to the phosphonates I, with 80-90%
yield, by oxidation with an hydrous t~butyl hydroperoxide ~0.25 Molly) according to H. Langhals, En Fritz end J.
Mergelsber~ Churn. Ben. 113~ 3662 (1980)) dissolved in I

acetonitr;le or tetrahydrofuran.
Alternatively, 30 my (0~95 Molly of sulfur are added to 0.7 Molly of VI at ~20C and the mixture is stirred overnight at room temperature. The reaction is generally already coTnplete after a few hours. 20 ml of chloroform are then added and the organic phase is extract ted three times by shaking with water. After drying over sodium sulfate and removal of the solvent, a crude product us obtained which is purified by silica gel chromatography to give the compound I on 80-90X yield table 6).
Alternatively, 118 my (1.5 Molly) of black selenium are added to 0.7 Molly of VI and the mixture us stirred overnight. After working-up (as above), the compound I
is obtained in 60% yield table 6).
I HPLC analysis of the reaction mixture (in the case where Z - 0 by comparison with the authentic reference Puff'. Two et at., Biochemistry 18, 5134 t1979~) showed clout 1% of the 3',3'~phosphonates and no 5',5' isomer.

~f~3~

c I
O I T
L I v Jo ., rJ
(L) E
Q , rJ O
Q O I I I I
Coo L N to I_ " O` V, TV TV V, TV V
to . O
I Us JO
l Us J Jo E o Ivy I It :-.
I- .. ' Jo ., X N
Q I
to I I-- I
I C . I N N N
I_ Jo r.
I I- ox --I O I 11 Ox 11 0 11 ~-( I I o -I
. . , -a aye . aye owe aye -a CO
O
ox ox ox o o . L I O JO
IIJQ D I . u . .
. I' I O .
I on Lo to l I- Q Q
l_ I
O` I

~,~ O D
I , , Jo Z I- v v ~'~
I- Cal I- r- co l l To To " I I
l l TV c to a I
X _ J I I (I) Jo V (_~ t; I TV

, I
'a Al .
. ,~` o I
I N I.) I O I I
I t-- (1) O I
Jo 11t~ --:~
N Z Jo O 2 I I Jo _ No-- N
Z N I_,_ N
r- -c I Jo I
-;~ N O I O l CO
. I I I. . .
N . O I I N o N .
ox) -c I 1 Jo 11 I N ED ill 11 N it to o I-- o E Al o , Jo Z I--N N
I`! I
'.'' I ~"'~ O O` N N
L-l I I I r- Jo or o I
I- I-- I) o r J ' JO Irk I CO
CLUE I- I-- . I- I_ ~~) JO 11~0 11 CO 11 ~11 I
-- -I to I- I-_.
N I N
Jo I
o Jo I ox N . N

Jo I I I
o o I- Ill Al r- Al o Al I
Jo I it rJ -Ox I_ I- I

I N N N
or- Jo ox Jo o -c It I
u- u us us J Jo N N N N
I Jo or_ . . . I, at:
I o us 11 11 it ox 11 us Lo I
I _ I so _ ox Cal r l . N r owe Jo I o I Jo AL N O` r-- Al o ox r I I l o o l l N l Jo No r ._~ I DUO I mu _ _ I, a Jo it -, 1 ' - 16 51~9 I N

N --N
N
`_ 00 r- if ~:~
Jo I
..
I I, M 7 Lo I Ox r JO
: T (Jo J _ O``- S S
ox a) C ,. __ I:
Q U
E . s s I--r~7 lo _ Jo L . 1'~1 I_ .. - C
I . 00 I_ Jo a `' --E I _ C
Q O . Jo E
I_ Lo C -- C O
~_~ _ CJ ill lo L
I co S I L JO Q. O
Jo I_ I C O :~:
I IO Jo a S JO
. E I I I o O
N r; ox ox m my L L. N Ill It It 01 N _ J
. -n tic L L S I
S Jo n I -o I

to N O
N I N l_ I O
Roy to I . .
It-- O It JO
_~~ If Jo O
Jo 1 1 7 0 I-- . r-l C
I- N 11 I I I . JO I It I: O` It IVY') E Jo U 11 N O I) . In Jo -a I. . Jo . Go . . --' I-- E
ox It J rJ
JO --I O` --' CO O O I (_> I-- I' . Z . LO 0 N O It I_ I --I I `-- Z t_ 0~~_1 Jo O JO arc Jo Q Al I- e- I_ I
L lo j ") I I It It I IO N N N N N N
-- O to \ :~: r I -I:: 3::
I I-- N U) Us OX CO i--L Jo two I JO 1 E
z I) I- I us If If Ill if If If a L '1: I-_ I- I- -I 'I
-O -O Jo L .

I ' O
V) ZOO I I _ o I US Al Us 1~1 O I I Al I if n.
r owe r . us I I us co o . .~_ I . TV
Lo It I_ It It I_ L_ I_ I Jo I I_ O E
Lo Ox O Ox I O C (n _. 1 It I I; I) I . Jo SUE -- --' `_ `-- E
I_ X E E E E C c -O
to to O O
E Jo Jo JO Jo to JO
r~J Al to Al r~J Al no foe O . D
Q L. a O Irk Us Us L L. I!
s o' I ox I Al o 3 i,_ I-, l I or n. or) o I l i l I IJoO

I
Al 1;1 . -I
I us us -1: Ill '. L
-- I I I Jo I JO _ Jo I (_~ ) I) to to - 18 - ~L235079 N
I I
a r-it N
JO
Jo O`
1. _ .
E
Q
Irk O
to It_ I
it I coy I I_ O ox En on .
r-CO
H
O`
In o I
I
O Jo E E
O ox O
I

" I C
a Q O
. Jo lo I I .
o .. ..
a I I_ N
I I N m CC
I

I g , .

N N N N
I: I I :1:
JO
US Us O
Us CO O I-- O`
n Al Al Al Al En Q

-- N l_ Q Q D, :1: Q
to Tao) (I) . OWE '' fly JO I' Jr/7 it I_> I-- --N Jo
2 Q Q C2. I Q
~17~ I L ox u) o co Jo Jo 00 I' I _, ED I_ L I_ I_ . .
a (IT O
V) 3 Q 3 0 'O O Q O Lo O
1~1 Jo J So J
O

x Jo E
. . ' Us JO
I
Of I E E to I_ Okay J
-- Us Jo I::
I) N

Jo I O
JO I
I
D. I r- in Q I) I J I
O Ill Q O O
I_ Us TV OF. .
I_ Ox (I I "I
. O O
l l l l l I) O_ Jo a Jo ._ J J
a X D

Claims (10)

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE PROPERTY
OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A Process for the preparation of deoxyribonucleoside phosphonates of the general formula I

(I) wherein T represents a protecting group for a primary hydroxyl group, B represents a nucleoside base radical in which any exoamino group present is protected, G represents a protecting group for a secondary hydroxyl group, Z represents oxygen, sulfur or selenium and R represents alkyl having up to 8 C atoms, cyclohexyl, benzyl or phenyl which may be substituted by fluorine, chlorine, bromine, lower alkyl, lower alkoxy or trifluoromethyl, wherein a difunctional phosphonylating reagent of the general formula II

(II) wherein X represents chlorine or Y and Y represents a group of the formula R1 and R2 representing identical or different alkyl or cycloalkyl radicals having up to 8 C atoms, or phenyl radicals, or R1 and R2, together with the nitrogen, representing a saturated or unsaturated heterocyclic ring which can contain further heteroatoms, is reacted with a nucleoside of the general formula III

(III) wherein T and B are as defined above, the resulting compound of the general formula IV

(IV) wherein T, B, R and Y are as defined above, is reacted with a com-pound of the general formula V

(V) wherein B and G are as defined above, and the resulting compounds of the general formula VI

(VI) wherein T, R, B and G are as defined above, are oxidatively con-verted to compounds of the general formula I.
2. A process as claimed in claim 1, wherein compounds of the formulae II to IV are used wherein T represents triphenylmethyl, p-anisoyldiphenylmethyl or di(p-anisoyl)phenylmethyl, B represents 1-thyminyl, 1-(N-4-benzoylcytosinyl), 9-(N-6-benzoyladeninyl) or 9-(N-2-isobutyroylguaninyl) and R represents methyl, ethyl, phenyl or benzyl.
3. A process as claimed in claim 1 wherein the compounds of the formulae II and III are reacted at -80 to +100°C.
4. A process as claimed in claim 3, wherein the reaction takes place at -20 to 0°C.
5. A process as claimed in claim 1, wherein the compounds of the formulae IV and V are reacted at -20 to +100°C.
6. A process as claimed in claim 5, wherein the reaction takes place at room temperature.
7. A process as claimed in claim 1, wherein the compound of the formula VI is oxidatively converted to compounds of the formu-la I at -80 to +100°C.
8. A process as claimed in claim 7, wherein the oxidation takes place at -20°C to room temperature.
9. A compound of the formula I

(I) wherein T represents a protecting group for a primary hydroxyl group, B represents a nucleoside base radical in which one exoamino group present is protected, G represents a protectings group for a secondary hydroxyl group, R represents alkyl having up to 8 C atoms, cyclohexyl, benzyl or phenyl which may be substituted by fluorine, chlorine, bromine, lower alkyl, lower alkoxy or trifluoromethyl, and Z represents sulfur or selenium.
10. A compound of the formula IV

(IV) wherein T represents a protecting group for a primary hydroxyl group, B represents a nucleoside base radical in which one exoamino group present is protected, R represents alkyl having up to 8 C atoms, cyclohexyl, benzyl or phenyl which may be substituted by fluorine, chlorine, bromine, lower alkyl, lower alkoxy or trifluoromethyl, and Y represents a group of the formula R1 and R2 representing identical or different alkyl or cycloalkyl radicals having up to 8 C atoms, or phenyl radicals, or R1 and R2, together with the nitrogen, representing a saturated or unsaturated heterocyclic ring which can contain further heteroatoms.
CA000462469A 1983-09-06 1984-09-05 Process for the preparation of nucleoside alkyl-, aralkyl- and aryl-phosphonites and -phosphonates Expired CA1235079A (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DEP3332068.3 1983-09-06
DE19833332068 DE3332068A1 (en) 1983-09-06 1983-09-06 METHOD FOR PRODUCING NUCLEOSIDALKYL, ARALKYL AND ARYLPHOSPHONITES AND PHOSPHONATES

Publications (1)

Publication Number Publication Date
CA1235079A true CA1235079A (en) 1988-04-12

Family

ID=6208339

Family Applications (1)

Application Number Title Priority Date Filing Date
CA000462469A Expired CA1235079A (en) 1983-09-06 1984-09-05 Process for the preparation of nucleoside alkyl-, aralkyl- and aryl-phosphonites and -phosphonates

Country Status (11)

Country Link
EP (1) EP0136543B1 (en)
JP (2) JPS6072899A (en)
AT (1) ATE38839T1 (en)
AU (2) AU570266B2 (en)
CA (1) CA1235079A (en)
DE (2) DE3332068A1 (en)
DK (3) DK162895C (en)
ES (1) ES8505384A1 (en)
GR (1) GR80287B (en)
IE (1) IE57904B1 (en)
PT (1) PT79172B (en)

Families Citing this family (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4959463A (en) * 1985-10-15 1990-09-25 Genentech, Inc. Intermediates
DE3916871A1 (en) * 1989-05-24 1990-11-29 Boehringer Mannheim Gmbh MODIFIED PHOSPHORAMIDITE PROCESS FOR THE PREPARATION OF MODIFIED NUCLEIC ACIDS
ATE151076T1 (en) * 1990-07-02 1997-04-15 Hoechst Ag OLIGONUCLEOTIDE ANALOGUES WITH TERMINALS 3'-3' OR 5'-5' INTERNUCLEOTIDE LINKAGES
US5512668A (en) * 1991-03-06 1996-04-30 Polish Academy Of Sciences Solid phase oligonucleotide synthesis using phospholane intermediates
AU678769B2 (en) * 1992-07-27 1997-06-12 Hybridon, Inc. Oligonucleotide alkylphosphonothioates
DE69402177T2 (en) * 1993-01-08 1997-06-26 Hybridon Inc SYNTHESIS OF DIMER BLOCKS AND THEIR USE IN THE COMPOSITION OF OLIGONUCLEOTIDES
US6087491A (en) 1993-01-08 2000-07-11 Hybridon, Inc. Extremely high purity oligonucleotides and methods of synthesizing them using dimer blocks
ES2086997T3 (en) * 1993-01-25 1996-07-01 Hybridon Inc OLIGONUCLEOTIDE-ALKYLPHOSPHONATES AND -ALKYLPHOSPHONOTIOATES.
US6417138B1 (en) * 1994-07-26 2002-07-09 Sony Corporation Method for transcribing an image and a support for transcription and ink ribbon employed therefor
EP0828749B1 (en) * 1995-05-26 2003-07-16 Genta Incorporated Methods for the synthesis of organophosphorus derivatives
CA2226111C (en) * 1996-05-03 2005-09-20 Hybridon, Inc. In situ preparation of nucleoside phosphoramidites and their use in synthesis of oligonucleotides
DE102004049339A1 (en) * 2004-10-08 2006-04-13 Basf Ag Process for the purification of phosphorus-containing chelate ligands
DE102007038930B4 (en) * 2007-08-13 2013-12-05 Universität Leipzig New chemical compound and its use in medicine, in particular for use in tumor therapy

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IL62834A0 (en) * 1980-05-14 1981-07-31 Ens Bio Logicals Inc Polynucleotide synthesis
US4415732A (en) * 1981-03-27 1983-11-15 University Patents, Inc. Phosphoramidite compounds and processes
EP0090789A1 (en) * 1982-03-26 1983-10-05 Monsanto Company Chemical DNA synthesis
DE3239888A1 (en) * 1982-10-28 1984-05-03 Hubert Prof. Dr. 2000 Hamburg Köster METHOD FOR PRODUCING OLIGONUCLEOSIDE PHOSPHONATES

Also Published As

Publication number Publication date
PT79172B (en) 1986-09-10
DK162895C (en) 1992-05-11
DK167359B1 (en) 1993-10-18
JPH0531560B2 (en) 1993-05-12
DK166585B (en) 1993-06-14
IE842268L (en) 1985-03-06
AU3275684A (en) 1985-03-14
IE57904B1 (en) 1993-05-05
DK131691A (en) 1991-07-05
EP0136543A3 (en) 1986-07-16
ES535627A0 (en) 1985-05-16
AU570266B2 (en) 1988-03-10
JPH0662662B2 (en) 1994-08-17
PT79172A (en) 1984-10-01
AU601257B2 (en) 1990-09-06
GR80287B (en) 1985-01-07
DK131591A (en) 1991-07-05
ATE38839T1 (en) 1988-12-15
DK131591D0 (en) 1991-07-05
JPS6072899A (en) 1985-04-24
ES8505384A1 (en) 1985-05-16
JPH05262787A (en) 1993-10-12
EP0136543A2 (en) 1985-04-10
DE3475307D1 (en) 1988-12-29
DK131691D0 (en) 1991-07-05
AU1138488A (en) 1988-07-14
DE3332068A1 (en) 1985-03-21
DK424784D0 (en) 1984-09-05
DK166585C (en) 1993-10-25
EP0136543B1 (en) 1988-11-23
DK162895B (en) 1991-12-23
DK424784A (en) 1985-03-07

Similar Documents

Publication Publication Date Title
CA1235079A (en) Process for the preparation of nucleoside alkyl-, aralkyl- and aryl-phosphonites and -phosphonates
US5552538A (en) Oligonucleotides with cleavable sites
US5367066A (en) Oligonucleotides with selectably cleavable and/or abasic sites
US5258506A (en) Photolabile reagents for incorporation into oligonucleotide chains
Dörper et al. Improvements in the phosphoramidite procedure for the synthesis of oligodeoxyribonucleotides
US5623068A (en) Synthesis of DNA using substituted phenylacetyl-protected nucleotides
EP0195277B1 (en) Phosphorylation process and agent therefor
WO1990008153A1 (en) Reagents for the preparation of 5&#39;-biotinylated oligonucleotides
EP0305201B1 (en) Protected biotin derivatives
US5166330A (en) Process for the preparation of nucleoside alkyl-aralkyl- and aryl-phosphonites and -phosphonates
DE69401136T2 (en) Modified oligodeoxyribonucleotides, their preparation and their therapeutic use
KR20030081303A (en) Synthons for oligonucleotide synthesis
Sawabe et al. Use of 2-trimethylsilylethyl as a protecting group in phosphate monoester synthesis
EP0144386B1 (en) Novel preparation of nucleoside phosphoramidite intermediates
CA2226111C (en) In situ preparation of nucleoside phosphoramidites and their use in synthesis of oligonucleotides
EP0295576B1 (en) Process for the production of the derivatives of 1,3,2-oxazaphosphorinane
US3005001A (en) O, o&#39;-(2-butenylene) and (2-butynylene) bis-(o-alkyl phosphoramidothioates)
JPH0588240B2 (en)
Cramer et al. Nucleotides, Part LX, Synthesis and Characterization of New 2′‐O‐Methylriboside 3′‐O‐Phosphoramidites Useful for the Solid‐Phase Synthesis of 2′‐O‐Methyloligoribonucleotides
CA1268173A (en) Preparation of nucleoside phosphoramidite intermediates
Dabkowski et al. Synthesis of phosphorofluoridates and phosphorofluoridothioates via the phosphoramidite approach
CA2088257A1 (en) Incorporation of selectably clevable and/or abasic sites into oligonucleotide chains and reagents therefor

Legal Events

Date Code Title Description
MKEX Expiry