JPS62212395A - Production of phosphorus amide compound - Google Patents

Abstract

PURPOSE:To efficiently obtain the titled bisamino type compound useful as a raw material for DNA synthetic reagents, by reacting a trihalogenophosphine with a secondary aminating agent and further reacting the resultant bisaminomonohalogenophosphine with an alcohol. CONSTITUTION:A trihalogenophosphine, preferably trichlorophosphine is reacted with a secondary aminating agent, e.g. diisopropylamine, etc., to synthesize a bisaminomonohalogenophosphine expressed by formula I (R1 and R2 are secondary or tertiary alkyl or both link to represent residue forming a heterocyclic amine; X is halogen), which is then reacted with an alcohol expressed by the formula R3-OH (R3 is OH-protecting group in phosphoric acid triester) to afford the aimed compound expressed by formula II.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to a method for producing a phosphorus amide compound, and more particularly to a method for efficiently synthesizing a phosphorus amide compound containing two secondary amino groups in the molecule.

(Prior art) With the recent development of genetic engineering, research has been actively conducted on methods for chemically synthesizing polynucleotides such as DNA (deoxyliponucleic acid) and RNA (liponucleic acid), which are the essential materials for 11L. It is.

Conventionally, methods such as the phosphodiester method, phosphotriester method, and phosphite method are known as chemical synthesis methods for polynucleotides, but among these, the phosphite method is attracting attention due to its reactivity. In particular, the amidite method using phosphoramidites is attracting attention.

In the case of the amidite method, chloro-N,N-dialkylaminomethoxyphosphine, chloro-N,N-dialkylaminocyanoethoxyphosphine, etc. are generally used as reagents for synthesizing nucleotide monomers (
For example, JP-A-57-176998, JP-A-60-50
2102, etc.), and previous studies have shown that the stability of nucleotide monomers produced differs depending on the type of substituent of the secondary amino group, and it has been found that the stability of nucleotide monomers produced differs depending on the type of substituent of the secondary amino group. It is known that those having a heterocyclic amino group are more stable than those having a chain substituent such as a dimethylamino group or a diethylamino group (for example, Tetrahedron Letters, VOL 24, A3, 245-248
Page, 1983).

Among these phosphorus amide compounds, bis-amino phosphorus amide compounds that do not contain halogens do not contain halogen atoms in their molecules, so when they are reacted with nucleosides to synthesize nucleoside phosphites, hydrogen halide is generated. It does not produce by-products and is convenient for operation.

(Problems to be Solved by the Invention) Therefore, the present inventors conducted intensive studies to develop an efficient method for producing a bisamino-type phosphorus amide compound useful as a raw material for a DNA synthesis reagent, and found that an inexpensive trihalogenophosphin The present inventors have discovered that a method using bisaminomonohalogenophosphine as a raw material is superior in terms of reactivity, and has completed the present invention.

(Means for Solving the Problems) Thus, according to the present invention, trihalogenophosphine and
After synthesizing bisaminomonohalogenophosphine represented by the following general formula (1) by reacting a class aminating agent,
There is provided a method for producing a phosphorus amide compound represented by the following general formula (111), which is characterized by reacting an alcohol represented by the following general formula [■].

(1) (u) (ll[
'1 (in the formula, R, and R2 each represent a secondary or tertiary alkyl group or a residue that forms a heterocyclic amine by combining the two, X represents a halogen atom, and R3Fil
It represents a protecting group for the hydroxyl group in the J-acid triester.

) In the present invention, first, a bisaminomonohalogenophosphine represented by the above general formula (1) is synthesized from a trihalophosphine and a secondary aminating agent.

Specific examples of trihalogenophosphines used include:
For example, trichlorophosphine, tribromophosphine, etc. are exemplified, and among them, it refers to secondary amines having the residue (R2 for economic efficiency is the same as above), gold amide, etc., and specific examples thereof include diisopropylamine, di- t-
Examples include butylamine, morpholine, thiomorpholine, pyrrolidine, piperidine, 2,6-dimethylpyrrolidine, piperazine, trimethylsilyldiisoglobylamine, trimethylsilyldi-t-ditylamine, lithium amide, sodium amide, aluminum amide, etc. Ru.

The reaction between trihalogenophosphine and a secondary aminating agent is
It is usually carried out in the presence of a solvent, and specific examples thereof include diethyl ether, tetrahydrofuran, benzene, toluene, and xylenato.

In the reaction, it is usually sufficient to use about 2 mol of the aminating agent per 1 mol of trihalor-nophosphine.
An aminating agent such as Yoshitado may be present for the purpose of removing hydrogen halide produced as a by-product during the reaction.

The reaction is usually carried out at a temperature of θ to 50° C. for about 2 to 30 hours, but is not necessarily limited thereto.

After the reaction is completed, bisaminomonohalogenophosphine is separated from the reaction solution. The separation means is not particularly limited, but it is usually carried out by distillation.

In addition, if a solid halide salt is produced as a by-product in the reaction solution, the solid content is separated in advance. However, in some cases, the next reaction may be carried out without separating the bisaminomonohalogenophosphine from the reaction solution.

In the present invention, the desired phosphorus amide compound represented by the above general formula (lit) is synthesized by reacting the thus obtained bisaminomonohalogenophosphine with the alcohol represented by the above general formula (II).

R3 in the above general formula (ill) may be any group as long as it acts as a protecting group for a hydroxyl group when a phosphotriester is formed in DNA synthesis. Specific examples thereof include a methyl group, an ethyl group, and a propyl group. β-cleavage of alkyl groups such as butyl groups, β-cyanoethyl groups, β-halogenoethyl groups, β-nitroethyl groups, β-thiocyanoethyl groups, β-methylsulfonylethyl groups, β-phenylsulfonylethyl groups, etc. Protecting groups that are removed by, allyl group, crotyl group, prenyl group, Grani/14
, cinnamyl group, p·-chlorocinnamyl group, and other allyl-type protecting groups.

For other specific examples of these protecting groups and methods of deprotection, see JP-A No. 57-176998 and Japanese Patent Application Publication No. 6
This method is disclosed in detail in Japanese Patent Application No. 0-502102, Japanese Patent Application No. 60-211240, etc.

Among these protecting groups, from the viewpoint of ease of deprotection and prevention of side reactions, protecting groups that are eliminated by β-cleavage and allyl-type film-retaining groups are preferred, and in particular β-cyanoethyl groups, carbon atoms of 6 or less The allyl-protecting group is recognized.

The reaction of the bisaminomonohalodanephosphine with the alcohol is carried out in a suitable solvent. An unusual example of a solvent is
Examples include diethyl ether, tetrahydrofuran, benzene, toluene, xylene, etc. When the bisaminomonohalogenophosphine is reacted without being isolated, it is desirable to use a common solvent.

In addition, other reaction conditions may be selected as appropriate; usually, 1 mole of the former is charged with 1-bil of the latter, and 2
This is done by reacting for ~30 hours. Further, it is preferable to coexist a base such as an amine in order to remove hydrogen halide produced as a by-product during the reaction.

Isolation and purification of the product from the reaction solution can be carried out by appropriately selecting or combining known methods such as distillation, adsorption chromatography, ion exchange chromatography, or distribution using organic solvents, which are common methods for organic synthesis reactions. It is possible to implement it by

(Effects of the Invention) Thus, according to the present invention, the desired phosphorus amide compound can be efficiently obtained using readily available raw materials such as phosphorus trihalodanide, a secondary aminating agent, and alcohol.

On the other hand, with the other possible method of reacting alcohol first, it is difficult to obtain monoalkoxydihalogenophosphine in good yield, and it is difficult to simultaneously react two secondary amino groups with bulky substituents. It is also difficult to introduce, and therefore the target product cannot be obtained efficiently.

(Example) The present invention will be described in more detail with reference to Examples below.

Example 1 After 28.6 mmol of phosphorus trichloride was dissolved in 3 QmJ of ether, 114.4 mmol of diisozolobylamine was added, and the mixture was stirred at room temperature for 20 hours. The reaction wave and the generated diisoderobyl ammonium chlorite were separated and distilled to obtain oily bis(N,N-diisozorobylamine)chlorophosphine in a yield of 70%. Furthermore, this screw (N, N
-diisozolobylamine) After dissolving 20 mmol of chlorophosphine in 30 m of ether, 2 mmol of triethylamine
0 mmol and 20 mmol of allyl alcohol were added, and the mixture was stirred at room temperature for 15 hours. After the reaction, the triethylammonium chloride produced was separated and the ether was distilled off, followed by distillation to obtain oily allyloxybis(N,N-diisopropylamine)phosphine. The total yield was 47%.

・Boiling 130-133℃/6■Hg ・HNMR (C6D6) 1.17 (dd, J-7,8,1,8Hz, 24H.

4 NCH(CH3)2 ), 3.53(d,
5ept, J-10, 8°7.8Hz, 41(
,4NCH), 4.10(dat, J = 1
0.5゜2 Hz + 2 Ht C=CCH2) +
5-07 (m, I H* c 1sCI(-
CHCH2'), 5.33 (m, IH, trans
CHCH2) -5,93 (ddt lJ” 1
8-10-5Hz lI H, CH2=CdanCH2
) ・”P NMR(Cb D b −Cb Hb , 1
: 4) 123-58 ppm Example 2 The reaction was carried out in the same manner as in Example 1 except that morpholine was used in place of diisopropylamine to obtain allyloxybis(modanphorino)phosphine. The total yield was 48 moles.

・Boiling point 145-155℃/0.6 wHg・HNM
R(C6D6) 2,90 (qlike, I, -, = JH-, =
4.5Hz, 8H.

4NCH2), 3.47(t, J=4.5Hz
, 8H, 400H2).

4.10 (m, 2H, C=CCH2), 5.07 (m,
IH,cisCdan=CHCH2)*5.3 0
(m e I Hr trans CH-CH
CH2), 5.88 (dat, p 1 shift 10
．． 5Hz.

IH, CH2-CHCH2)-”P NMR (C6D6-C6H6,1:4)
130.45 ppm Example 3 The reaction was carried out in the same manner as in Example 1 except that β-cyanoethanol was used instead of allyl alcohol to obtain β-cyanoethoxybis(N,N-diimpropylamino)phosphine. . The total yield was 42 moles.

・Boiling point 123-130℃/0.01 mHg-P-N
MR (C6D6-041 (801: 2) 123.
85 ppm-H-NMR (CD, Ct) 1.35 (d, 24H4NCI ((CMp2)
3,82 (m, 4 H4NCH(CH3)2)4
．． 03.4.25 (2t, 2H-0□2CH2CmiN)
2.80 (t, 2H-0CH2%CmiN) Comparative Example 1 After dissolving 30 mmol of phosphorus trichloride in 30 mKiJ of ether, 30 mmol of β-cyanethanol and 30 mmol of pyridine were added and stirred at room temperature for 20 hours. After the reaction, the produced → lysinium chloride was separated and distilled to obtain oily β-cyanoethoxydichlorophosphine. The yield H was about 151.

Without this β-cyanoethoxydichlorophosphine 20
After dissolving 30 mmol of ether, 80 mmol of twinzolobylamine was added and the mixture was stirred at room temperature for 20 hours. However, the desired β-cyanoethoxybis(diimpropylamino)phosphine was hardly obtained.

Claims (1)

[Scope of Claims] 1. After synthesizing bisaminomonohalogenophosphine represented by the following general formula [I] by reacting trihalogenophosphine with a secondary aminating agent, an alcohol represented by the following general formula [II] is synthesized. A method for producing a phosphorus amide compound represented by the following general formula [III], which comprises reacting. ▲There are mathematical formulas, chemical formulas, tables, etc.▼[I]▲There are mathematical formulas, chemical formulas, tables, etc.▼[II]▲There are mathematical formulas, chemical formulas, tables, etc.▼[III] (In the formula, R_1 and R_2 are respectively secondary or a tertiary alkyl group or a residue in which both are bonded to form a heterocyclic amine, X represents a halogen atom, and R_3 represents a protecting group for the hydroxyl group in the phosphoric triester.)