JPS60260586A - Preparation of phosphoric acid triester compound - Google Patents

Abstract

PURPOSE:To obtain the titled compound useful in the fields of phospholipid and nucleic acid chemistry, at a high rate of reaction, by reacting a diorganophosphoro-triazolide compound with an organotin alkoxide compound in the presence of a specific amine. CONSTITUTION:The objective compound of formula II can be produced by reacting (A) diorganophosphoro-1,2,4-triazolide compound of formula I (R<1> is alkyl or aryl; R<2> is alkyl, aryl, or N,5'-protected deoxynucleoside) with (B) organotin alkoxide compound of formula R<3>3SnOR<4> (R<3> is same as R<1>, R<4> is R<2> or N,3'- protected deoxynucleotide) in the presence of (C) an amine of formula HNR<5>R<6> (R<5> and R<6> are alkyl or R<5> and R<6> together with adjacent N form piperidino which may have alkyl-substituent group) (e.g. 2,2,6,6-tetramethylpiperidine, etc.).

Description

DETAILED DESCRIPTION OF THE INVENTION This invention relates to an improved method for producing phosphotriester compounds which are particularly useful in the fields of pesticide, phospholipid and nucleic acid chemistry.

BACKGROUND OF THE INVENTION First of all, a diorganophosphoro-1,2,4-)')azoli)' compound represented by the general formula [I], an organotin alkoxide compound represented by the general formula (10), and an organotin alkoxide compound represented by the general formula (10) It was discovered that the following reaction formula 411 K Yotsu [reacted to give a phosphoric acid triester compound [[ ]
-233485 and the specification of Japanese Patent Application No. 1982 filed on June 7, 1982).

1 (R' 0) (R20) P (OR' ) (1) [
1) (in the formula) R is an alkyl group or an aryl group, R2 is an alkyl group, an aryl group, or N, 5'-protected dinucleoside'b R is an alkyl group or an aryl group,
R is an alkyl group, aryl g, N, 3'-ffl-protected dinucleoside, or N, 3'-protected dinucleotide. ) This reaction is different from the conventional method for producing phosphoric acid triester compounds, in which an organic phosphorus compound is activated with a catalyst, etc., and this active species is reacted with an alcohol. It is activated and reacted with an organic phosphorus compound to produce a phosphoric acid triester compound.

According to this method, it is generally possible to produce a phosphoric acid triester compound [degree] with a high degree of stability, but at this time, the organic group R4 of the compound [[) has a sterically bulky structure such as a t-butyl group. In the case of such a group, there was a problem that the reaction rate was not large, such as t. Considering this situation, the present inventors conducted various studies with the aim of finding a factor that promotes reaction (1). . As a result, they discovered that the objective could be achieved when the reaction was carried out in the presence of an amine, and completed the present invention. In the present invention, a diorganophosphoro-1,2,4-') riazolide compound represented by the general formula CI] and an organotin alkoxide compound represented by the general formula [II) are reacted to form a compound of the general formula [III]- 1: When producing gold from the phosphotriester compound shown, the reaction is carried out according to the general formula [
■] I Blood RfL [IV) (In the formula, R5 and R6 each represent an alkyl group, or both together together with the adjacent silicon atom represent 1
A phosphoric acid triester compound represented by the general formula [III], which is characterized in that the reaction is carried out in the presence of an amine represented by (including a piperidino group optionally having one or more alkyl substituents) The present invention provides a method for manufacturing.

In the method of the present invention, 2 represented by the above general formula [IV]
grade amine is used to promote the reaction (11),
Generally, it has been recognized that the larger the acid dissociation constant (pKa), that is, the more basic the amine is, the more effective it is in promoting the reaction (1). Specific examples of such amines include:
Di-n-propylamine, di-l--/ropylamine, di-n-butylamine.

Di-l-butylamine, di-t-butylamine.

Piperidine, 2,6-dimethylpisridine.

2.2.6.6-titramethylpiperidine and the like can be mentioned.

These amines can be produced by known methods or commercially available, but it is preferable to use amines that have been sufficiently dehydrated on a drying side appropriate for the music to be used.

The organic groups R and R of the diorganophosphoro-1+14-)IJDing zolide compound represented by the general formula CI), which is the starting compound in the present invention, may be either an alkyl group or an aryl group, but in some cases, R may be It may also be an N,5'-protected dinucleoside containing an amide group or a carbonyl group, as described below. Compound CL) is the reaction of a diorganophosphorochloride compound represented by general formula [V] with 41(-1,2,4-triazole) in the presence of a base such as triethylamine. It can be produced by the reaction formula (2)) or the reaction of the organophospho-a-di-1,2,4-triazolide represented by the general formula [■] with an alcohol (reaction formula (3)).

(VI) (In the formula, R and R have the same meanings as above.

) The compound of the general formula [■] can also be produced by the reaction of the diorganophosphorochloride compound [v] and the triorganostannyl-1+2+4-t'J asoru represented by the general formula [■] (Reaction Formula (4)) %Formula% (In the formula, at and R2 have the same meanings as above.

R represents an alkyl group0) Which of the above three methods to use for producing compound [I] depends on the properties of the organic groups R and R2 or the starting materials of compounds [v] and [VI). Depending on availability etc., reactions (2) and (4) are generally carried out when R and R are an alkyl group or an allinyl group, and when Ft+ is an alkyl group or a 7-aryl group and R2 is N
, in the case of 5'-secured dexoxynucleosides, the reaction (
It is convenient to use 3).

Next, we will explain about the alkoxytriorganostannane represented by the general formula C [ ), which is another important raw material of the present invention. The substituent R may be an alkyl group or an aryl group, but the compound [■] When considering the case of isolation and purification, lower alkyl groups such as methyl group, ethyl group, or n-butyl group are preferred because they are easy to purify.Also, organic groups &4
Examples include alkyl groups such as methyl, 11-propyl, and t-butyl, as well as N, 3'-protected deoxynucleosides containing an amide group and a carbonyl group, as will be explained later. As already known, methods for producing compound [II] include a method in which a triorganotin chloride compound and a corresponding metal alkoxide compound (MOFL') are reacted, and a method in which a triorganotin chloride compound is reacted with a corresponding metal alkoxide compound (MOFL');
) as shown in the triorganostinamide compound [II]
There is a method by reacting with the corresponding alcohol ()IOR').

R8nNRR+HOR4R8nOR+ HNR'R'
(513 [III [II) (wherein R3 and R4 have the same meanings as above, and Re and R9 have an alkyl group 'kl0) According to reaction (5), compound [III and alcohol ( I(O
By simply reacting R') in an organic solvent such as methylene chloride, chloroform, or ether at room temperature for about 5 to 10 minutes, the compound [''] can be obtained quantitatively, and the amine compound (ENFL'R'') produced as a by-product can be obtained quantitatively. Since this does not interfere with the next reaction (reaction (1)), compound (III) can be used in the form of methylene chloride, chloroform, or ether acid 1 in a so-called "in situ" manner, which simplifies the production reaction operation. There are advantages.

As mentioned above, according to the present invention, by adding an amine of general formula CW3 to reaction (1), the reaction rate can be significantly increased. For example, compound (I) is 5
'-0-dimethoxytrityl-thymidine-3'-0-
(2-chlorophenyl)phosphoro-IT 21 4-
to! j azolide υ, and the compound [[] is 5'-0-)ethylstannyl-3'-0-t-butyldimethylsilyl-thymidine, the reaction completion time is about 30 minutes, but this When diimprobilamine is added to the system, the reaction is completed in about 5 minutes. Compound (I) is dimethylphosphoal.

2.4-)! jγzolide, and compound (II) is i
-propoxytriethylstannane, the reaction time is about 5 minutes, while in the presence of 2.2°6.6-titramethylpiveridine, the reaction time is about 1 minute.
shortened to minutes. Furthermore, the compound WdsCI] is diphenylphosphoH-1,2,'4-triazolide,
When the compound [■] is t-butomycintriethylstannane, the time required for reaction (1) to complete is about 20 hours, but when 2,6-dimethylpi is added to this reaction system, it reacts. The time is reduced to about 3 hours. In this way, amines [
By adding [IV], the reaction completion time can be shortened to 175 to 1/10 compared to a system without additives. is the reaction (2), (3) or (
4) or organophospho C! -1°2.4
-) an organic tin alkoxide compound represented by the general formula [II] in the lyazolide compound [■], and then an organic tin alkoxide compound represented by the general formula (IV)
It can be produced in a short time by adding the amine shown below. It is preferable to carry out this reaction in an organic solvent such as methylene chloride, chloroform, or benzene.It is better to thoroughly dehydrate these solvents with a desiccant suitable for the song being used. Although the reaction can be carried out at 0 to 35'C, it is convenient to carry out the reaction at room temperature, which does not require operations such as cooling and heating. Compound [1) and compound (II)
The molar ratio of the organic groups R'+ may vary somewhat depending on the properties of R2 and R4, but in general it may be 1:I.

Further, the molar ratio of compound [II) and amine [■] is usually 1:
1 may be used, but in the case where the organic group B in the compound (10) is an N, 3'-protected dinucleoside or an N, 3'-protected dinucleoside or cleotide, it is preferably from 1:5 to 2011 degrees.

Since the reaction completion time differs depending on the properties of the organic groups R, R, and B, it is best to check the progress of the reaction by gas-chromatography, thin-layer chromatography, or HNMR, and then perform KWk treatment. preferable. Isolation and purification of the phosphoric acid triester compound - [■] can be carried out by distillation, thin layer chromatography or column chromatography, but generally the compound (I [I
Dinucleoside in which at least one of the above organic groups is N=5'-protected dinucleoside, N, 3'
In the case of -protected deoxynucleotides or N,3'-protected thionucleotides, thin-layer cymography or column chromatography may be used.

The structural formulas of the N,5'-protected deoxynucleoside, N,3'-ff1M deoxynucleoside P and N,3'-protected deoxynucleotide described herein are as follows.

Thymine residue N-hendiylcytosine residue N6 -Benzoyladenine residue N2-Imbutyrylguanine residue DMTr: Dimethoxytrityl group 00CH3CH1 Dinocetyl group, benzoyl group, t-butyldimethylsilyl group phenyl group, 2-chlorophene 4.4-chlorophenyl, group n: positive integer.

EXAMPLES The present invention will be fully explained in detail with reference to Examples below, but the present invention is not limited to these Examples.

IJ! Example 1 1 (' -P rO) P (OMe) 2 [■'] Dimethyl chlorophosphate and 41 (-1,2゜) obtained by a known method from 4-trichozole and triethylamine or methyl phosphoro-1,2 ,4-triazolide [■
'] in ether solution (0.8M.

6.25 at, 5 mmol) in 1-propo citriethylstannane CL3 (1,325 f, 5 mmol)
)-ek)vi- and immediately add 2.2.6.6-titramethylpiveridine clear'] (0.706 r, 044 m,
5 mmol) was added. After 1 minute, acetone (20d)
was added, and after separating the produced white solid fP, the p liquid was distilled to obtain a distillate with a boiling point of 73-74°/9 planes old t of 0.67
32 Obtained Kokomono Co. Dimethyl-1-probylphosph
E-to Cm'l (U, Fe1eh mr M,
R@gitz, kngew.

chem, +ε8.377 r1976)), and the yield was 80-. ')INMa (CDC/
3. TMS) δ: 1.34 (a.

6H, J = 7. OT(Z, (C!!3)2CHO)
, 3.68 (d, 6H.

Jz12. OHz, CH30), 4,30-4.80(
m, IHl(CH3)2CQO) ppm.

1 (t-BuO>P(OPh)2 [■'] diphenylchlorophosphate and 4) (-1,2゜4-
To a deuterated chloroform solution (IM, 2 m, 2 mmol) of diphenylphosphoro 1゜2.4-) riazolide [I'] prepared by a known method using triester and triethylamine, t-ptocytriethylstannane (II) was added. '
l (0,558f, 2 mmol) and 2,6-dimethylbilysine (iV') (0-2269.

0.27 ml (1.2 mmol)'t- was added. 3 hours later
't(NMR spectrum of this solution is diphenyl-t-
Butyl phosphate [IJf] (G, Goanovs
ky, starvation I (.

Zaet, W-Scbmltt+ J-Org-Cbe
m, 35, 336 (1970)) was produced in a yield of 95%. Add this solution to silica gel (
Column croftography of 5t) was carried out and developed with 2 solvents of aroform-methanol (+o:+) to obtain 0.55% of diphenyl-t-butyl phosphate [■'].
151 (yield 9Q%) was obtained. 'r(NMR(CbC1
3, TMS) δ: 1.54 (IJ, 9H2rcu,
), 0), 6+85'-' L35 (rn, IO'F
I.

Ph) ppm.

Example 3) 1'N (CI' to 1-P) [['] (where DMTr is a dimethoxytrityl group, T is a thymine residue, TBDMS is a t-butyldimethylsilyl group, and EL is an ethyl group, i-r'r means i-propyl group) 5'-〇-dimethoxytritylthymidine and 2-cI
is phenylphosphorology 1, 2, 4-)
5' produced by a known method from IJ Ryozolito °
-0-dimethoxytrityl-thymidine-3'-0-(2
-Riocyfuconinyl)phosphoro-1,2,4-to!
methylene chloride solution (o, gM, Q
,3m+0.24 mmol)'k,(diethylamino)triethylaminenan(0,058y,o,os
mjt, 0.21 mmol) and 3'-0-t-butyldimethylsilyl-thymidine (0,071?, 0.2 mmol) were stirred in mef'I/n chloride (0-2 ml) for 15 minutes. Produced 95'-〇-triethylstannyl-3'
-o-t-butyldimethylsilyl-thymidine diethylamine salt [''] was added at 20'C. Then immediately diimpropylamine [iv') (o, to + t,
0. After confirming the completion of the reaction by thin layer chromatography (hereinafter abbreviated as TLC), 50 tipyridine aqueous solution (0.5d) was added. This was washed with a 0.5M aqueous potassium phosphate solution (low/) and 0.1M triethylamine bicarbonate (PH7.5, 10TWt), and then distilled off under reduced pressure in a methylene chloride chamber.
The residue was subjected to Lof column chromatography and eluted with chloroform-methanol (40:1). The eluate was concentrated, the residue was dissolved in a small amount of methylene chloride, and the solution was added dropwise to vigorously stirring n-hegisan, and the resulting white crystals were separated from P and dried to give 5'-〇-dimethoxytrityl-thymidyl. (3'→5')-3'-0
-t-butyldimethylsilyl-thymidine-3'-0-(
0.204 ri of Pl-2-chlorophenyl)phosphate) [[').

Yield: 95% (vs. 5'-0-)ethylstannyl-3
'-0-t-butyldimethylsilyl-thymidine), and its physical properties were as follows.

Thin layer chromatography (TI, C), R-0,6
7 (CHC7!3:MoOH=9:I). 'H
NMR (CDL) 7!3 r TM8) δ: 0
．． 0B (s, 6T(,(C)+3)28I), 0
．． 90 (s, 9T(, (CI(3)3C8I),
1.14 (fi, 3H,5-CI5).

1.85 (s, 3H25-C!!3), 2.0-2.8
0 (m, 4H92' and 2'), 3-20 3.
60 (m, 2H95').

3.80 (s, 6H2C!!30 and C horse 0), 3
．． 90-4・70 (m, 5H, 3', 4',
4' and 5'), 5.30-550 (m, IH
, 3'), 6-29 (t, It(, J = 7,0f
(Zl l')l 6,50 (t, IHI J =7
．． Of (z, I') 16-80 7.00 (nl+
4H, Ph ), 7.10 7.80 (m.

+5H, 6, 6 and ph), 9°72 (g, 2T
(, NU and NH) ppm.

3'-0-phospho0-1.2.4-) of deoxynucleosides listed in Table 1; 5'-0- of tothioxynucleosides or dideoxynucleotides;
) The same operation as in Example 3 was carried out using the amine salt of the liorganostannyl derivative.
The meanings of the abbreviations in Table 1 of il90 are as follows.

pJ: x thyl group, n-Bu; n-butyl group, 1-pry
s-propyl group + DMTr * dimethoxytrityl group, Bz; benzoyl group, TBDM8; t-butyldimethylsilyl group, T; thymine residue, ClN4-benzoylcytosine residue Mr A''; N6-pensoyl adenine residue cib; N2 - Imptirylgaryinyl residue.

87

Claims (1)

[Claims] Diorganophosphoro-1゜2. represented by the general formula CI).
4-) Reacting the lyasiliP compound and the organotin alkoxycyto compound represented by the general formula [■] to form the general formula [11[)
In producing a phosphoric triester compound represented by the general formula [IID], the skeleton reaction is carried out in the presence of an amine represented by the general formula [■]. Method 〇〇 (In the formula, R' is an alkyl group or an aryl group, R is an alkyl group, an aryl group, or N, and 51-protected thio dinucleoside.) R8nOR (In the formula) R is R4 is an alkyl group or an aryl group, R4 is an alkyl group, an aryl group, N, 3'-protected deoxynucleoside or f-JN, 3'-protected deoxynucleotide 1e, 0) 1 (a o) (a o ) p (oa) (m) (s in the formula
R' * R' and R4 have the same meanings as the sergeant. ) HNRR [ff) (wherein R and R each represent an alkyl group, or both together with the adjacent nitrogen atom may carry one or more alkyl substituents. Hail.)