JPS61205296A - Anhydrophosphononucleoside derivative and production thereof - Google Patents

Abstract

NEW MATERIAL:The compound of formula I (R<1> and R<2> are H or lower alkyl). EXAMPLE:2, 3'-O-anhydro-1'-diethylphosphono-4', 6'-O-(tetraisopropyl-disiloxane-1, 3-diyl)-BETA-D-fructofuranosyluracil. USE:An agent for suppressing the biosynthesis of nucleic acid. It is useful also as a carcinostatic agent and antiviral agent, etc. PREPARATION:The compound of formula II (R<3> is OH-protecting group; R<4> is lower alkyl) [e.g. 2, 3'-O-anhydro-1'-methoxy-4', 6'-O-(tetraisopro phldisiloxane-1, 3-diyl)-beta-D-fructofuranosyluracil] is made to react with the compound of formula HP(O)(OR)2 (R is lower alkyl) (e.g. diethyl phosphite) in the presence of a base (e.g. triethylamine), preferably in an ether solvent (e.g. THF) at 0-55 deg.C, and the product is deprotected to obtain the objective compound.

Description

Detailed Description of the Invention [Industrial Application Field] The present invention relates to a novel anhydrophosphono compound having the general formula (1) (wherein R and R are the same or different and represent a hydrogen atom or a lower alkyl group). Concerning nucleosides and their production methods.

More specifically, it relates to the anhydrophosphononucleoside derivative represented by the general formula (1), which is a nucleic acid biosynthesis inhibitor and is therefore expected to develop in the future as an anticancer agent and an antiviral agent. be.

[Conventional technology]

Many nucleic acid biosynthesis inhibitors have been synthesized, and 6-merkatoprine is well known as a sybrin-based biosynthesis inhibitor, and 5-fluorouracil is well-known as a pyrimidine-based biosynthesis inhibitor.

Among phosphononucleosides, a large number of 5'-phosphonate compounds have been synthesized, including 5'-deoxy-5'-phosphonouracil (
A, Hol'y T@trahsdron Le
tt@rs.

881 pages 1967). However, there is no description of a nucleoside having a phosphono group or a phosphonomethyl group at the 1' position.

[Problems that the invention attempts to solve]

Many efforts have been made to discover nucleic acid biosynthesis inhibitors and apply them to anticancer and antiviral drugs. However, among the pyrimidine biosynthetic pathway, orotate phosphorigesyl transferase (0rotate phosphorigesyl transferase)
The nucleobase and phosphoryl by holboayl Transferase) are converted into sylpyrophosphate (Pho
sporlbosyl pyrophosphate
) Studies on competitive inhibitors corresponding to the transition state in the condensation step have not yet been conducted.

The present inventors planned to synthesize a transition state-like compound as a competitive inhibitor in which phosphoryl replaces the oxygen of the carbon-oxygen-phosphorus bond at the 1st position of silpyrophosphate with carbon and introduces a nucleobase at the 1st position.

These biosynthesis inhibitors are expected to be used not only as anticancer agents and antivirals, but also as central nervous system agents and agricultural chemicals, such as insecticides.

[Means for solving problems]

The present invention discloses a novel compound in which orotate phosphorylate is a competitive inhibitor of the transition state in the condensation step of sylpyrophos 7ate with a nucleobase formed by syltransferase, an anhydrophosphonolytic compound having the following general formula (1). The anhydrophosphononucleoside derivative of the present invention, which is a nucleoside derivative and its production method, is prepared by the following method. Can be synthesized.

That is, the known substance 2,3'-anhydro-β-D-7 lactofuranosyluracil (A, Holy et al. Nuclsic Aci
d Res a, vol. 1, p. 289, 1974)
was used as the starting material.

In this compound (3), the hydroxyl groups at the 4' and 6' positions of the sugar are selectively protected to form a compound (4) (wherein R represents a protecting group). The protecting group used here may be any one that selectively protects the 4' and 6' positions of the sugar, but includes silyl-based protecting agents,
In particular, dichlortetraisofurovir disiloxane is preferably used.

By oxidizing this protected compound (4), an aldehyde isoside having the general formula (5) (wherein 85 represents the same meaning as above and R4 represents a lower alkyl group) is obtained.

The oxidizing agent used here is preferably dimethyl sulfoxide, such as dimethyl sulfoxide and trifluoroacetic acid/pyridine/dicyclohexylcal diimide, dimethyl sulfoxide and phosphoric acid/dicyclohexylcardiimide, and dimethyl sulfoxide and acetic anhydride. Further, the reaction temperature is preferably room temperature to 50°C.

In this oxidation reaction, the aldehyde product is easily converted into a hydrate form, and reacts with the solvent used in the purification process, especially alcohol, to obtain the aldehyde equivalent represented by the general formula (5). Then, by using silica gel column chromatography and eluting with methylene chloride/methanol, the group R becomes a methyl group.

This aldehyde equivalent (5) and general formula (6)% formula% (6
) (wherein R represents an alkyl group) is reacted in the presence of a base to obtain a phosphononucleoside having the general formula (2) (wherein R and R represent the same meanings as above). It will be done.

Here, the base is a trialkylamine, such as triethylamine or diimpropylethylamine.

Examples include triisopropylamine and metal alkoxides such as sodium methoxide, sodium ethoxide, potassium ethoxide, and potassium t-butoxide. Also suitable is sodium hydride.

For example, dibenzyl phosphite and diphenyl phosphite (11) are substituted for dialkyl phosphite, and each group R
A compound in which is a benzyl group or a phenyl group is obtained.

In this reaction, the reaction temperature is preferably 00 to 55°C, and the best solvent is ether or alcohol.

A general deprotection reaction is used to remove the protecting group of the compound represented by formula (2) to obtain the compound of the present invention. For example, in the case of a silyl-based protecting group, deprotection can be performed using a general tetraalkylammonium 70lide.

The reaction solvent may be any solvent that does not participate in this reaction, and the resulting compound having the general formula (1a) (wherein R has the same meaning as above) can be processed by column chromatography, preparative thin layer chromatography, and reprocessing. It can be purified using methods such as crystallization.

In addition, when the group R in this compound (1a) is a methyl group, t-
Treatment with butylamine yields an anhydrophosphononucleoside derivative having formula (1b).

In this reaction, the reaction temperature may be 40 to 80°C, and if one of the groups is a methyl group, only that methyl group is hydrolyzed.

When the anhydrophosphononucleoside derivative having the general formula (1a) is treated with trimethylsilyl iodine in a chlorinated solvent, it becomes a compound having the general formula (IC); when it is treated with an acylating agent, it becomes a compound having the general formula (1d) ( It is obvious to those skilled in the art that a compound having the following formula (wherein R has the same meaning as defined above and A represents an acyl group) can be obtained.

Next, the present invention will be explained in more detail with reference to Examples, but it goes without saying that the present invention is not limited to these Examples.

Example 1 2.3'-0-anhydro-1'-diethylphosphono-
4',6'-0-(tetraisopropyldisiloxane-
1,3-diyl)-β-D-fructofuranosyluracil Compound 264 of Reference Example 2 was dissolved in 2114 of tetrahydrofuran, and 276rn9 of diethyl phosphite (
4 equivalents) and 0.281117 triethylamine (4 equivalents) were added and stirred for 1 day and night. The reaction solution was concentrated and the residue was subjected to column chromatography on silica gel (50P) and eluted with methylene chloride/methanol (95:5).
The title compound I#9 (94% yield) was obtained.

This compound had two types of isomers regarding hydroxyl group and phosphorus, and the ratio was 65:35.

Physical properties of main product UV (ethanol, 2ml LX s nm
): 247 (8950), 226 (10300) IR
Suctor (KBr, on): 1660,15
50,148ONMR d Tor (CDC2,, δp
pm): 0.8-1.1 (my281), 1
．． 37 (t, 6H, J=6.9Hz), 3.9-4.0
(m.

2H), 4.15-4.45 (m, 6H), 5.08 (
dd, IH, J-12, 5, 7, 9Hz), 5.75 (
d, IH, Jx5.6Hz), 5.88 (ct, I
H, J-7, 11'h), 6.86 (brdd,
IH, J = +14.4, 7.9Hz), 7.70 (d,
IH, Jm7. IHz) Physical properties of the other isomer NMR spectrum # (CDC1, *δppm): 0.8
~1.1 (m.

28H), 1.38 (t, 6H, J-6°9Hz), 1
．． 75 (br.

IH), 3.90-4.05 (m*2H)s
4.15-4.45 (m.

6H), 5.18 (d, IH, J-12, 7Hz), 5
．． 81 (d.

IH, J-5, 6Hz), 5.91 (d, IH, Jx7
．． IHz).

7.38 (d, IH, J = 7.1 Hz) Example 2 2.3'-0-a/hydro-1'-dimethylphosphono-
4',6'-0-(tetraisopropyldisiloxane-
1,3-diyl)-β-D-fructofuranosyluracil Compound 400rn9 of Reference Example 2 was dissolved in 4 Qiu of tetrahydro7 run to give dimethyl phosphite (4
equivalent) and 0.43111 triethylamine (4 equivalents)
was added, and the reaction was carried out in the same manner as in Example 1. Purification was carried out using silica gel column chromatography and elution with methylene chloride/methanol (9:1) to obtain the title compound 4181Q (yield 89Q).

This compound had two types of isomers regarding hydroxyl group and phosphorus, and the ratio was 3:2.

The physical properties of the main product are shown below.

Properties: Amorphous crystal Uv spectrum (ethanol, λmaxtnm): 24
7 (8670), 226 (10100).

IR spectrum, v (KBr, m): 1660, 15
40,147ONMR soukt/l/ (CDC1,,
629m): 0.8-1.1 (m.

28H), 3.7-4.0 (m, 8H), 4.3-4.
4 (m, 2H).

5.17 (dd, IH, J-8, 2, 13, 2Hz),
5.67 (d.

I Hy J-4-OHz ) * 5-84 (d
s I H-J-7,8Hz) +6.97(dd,
IH, J”8.2, 15.2t(z), 7.68(d.

IH, J-7,8Hz) Mass spectrum (m/z): 563 (M-43) Physical properties of the other isomer: Amorphous crystal NMRx dct A/ (CDC1,, δppm):
5.71 (d, IH.

J 4-OHz) -5,85(d*IH
s J 7-8 Hz) s 6-97 (dd, I
H, J = 8.2, 15.2 Hz), 7.43 (d, IH
, J-7,8Hz) Example 3 2.3'-0-anhydro-1'-dimethylphosphono-
β-D-7 Lactofuranosyluracil Compound 185Iv of Example 2 was dissolved in 1 lau of tetrahydrofuran, and a solution of tetra-n-butylammonium 70 lide in tetrahydrofuran (1 molar solution, 9
．． 51 m/) was added and stirred for 30 minutes.

After adding 3 drops of water to the reaction solution and decomposing it and concentrating it, the residue was subjected to column chromatography using silica gel (20P) and eluted with methylene chloride-methanol (80:20).
8 parts of the title compound were obtained (yield: 97 parts).

This compound had two types of isomers regarding the hydroxyl group at the 1' position of the sugar, and the ratio was 3:2.

Physical properties of main product: Properties: Amorphous crystal [α) knee 33.1° (methanol) UV spectrum (ethanol, λm□@n!n)
:248(#-8080), 225(#tomi9180)I
R spectrum (KBr, am): 3500,166
0°1540.148O NMR spectrum (CD, OD, δppm): 3.
61(d, 2H.

J wI3.4 Hz ) * 3.88 (d -3
H, J ■11-OHz) -3,92(d, 3H
, J-10.3Hz), 4.39 (dt, IH, J-2
,6Hz),4.56(dd,11.J-1,
9,2,6Hz), 4.77.

(d, IH, J-11, 5Hz), 5.58 (d, IH
, J-1,9Hz)6.12(d,IH,J=7.2H
m), 7.98 (d, IH, J-7,2Hz) Mass spectrum (m/z): 281 (M”-83) Physical properties of the other isomer: NMR spectrum/I/ (CD, OD, δp pm
): 3-57 (d e 2 HsJ -3,8H
z ) s 3.72 (d * 3 Ha J ; 1
0-8 Hz) s 3.86 (d, 3H, J=10
．． 1Hz), 4.32 (dt, IH, J=3.8°3.
6Hz), 4.52(dd, IH, J-1,
9, 3, 6Hz), 4.68 (d, IH, Jxll
, IHz), 5.42 (d, IH, Jxl, 9Hz).

6.03 (d, IH, J-7, 4Hz), 8.02 (d
, IH, J-7°4Hz) Example 4 2.3'-0-7hydro 11-methylphosphono-β
-D-Fructofuranosyluracil H The main product 78.6~ of Example 3 was dissolved in 3.51 t-testeramine and heated under reflux for 8 hours. The reaction solution was concentrated, the residue was dissolved in water, and subjected to ion exchange chromatography (DOW).
We visited @X50WX4). By eluting with water and concentrating the eluate, the title compound with fi74.3# (yield 98%) was obtained. there were.

Physical properties of main product: Properties: Amorphous crystal UV spectrum (ethanol tλmax*nm): 26
2 (#-5510) r Rs 4 r) le (KBr, all-'): 3400,
1690°148O NMR quictor (CD30D, δPP!n):
3.5-3.7 (m.

2H), 3.76 (d, 3H, J-10,9Hz), 4
．． 36-4.41 (m, IH), 4.52-4.59 (
m, 2H), 5.58 (d, IH.

J=1.6Hz), 6.13(d, IH, J=s7.5
Hz), 8.10 (d.

IH, J=7.5Hz) Mass spectrum (m/z): 225 CM"-1
25) Physical properties of the other isomer: NMR spectrum (CD, OD, δP Pm):
3-5 to 3.7 (m t2H), 3.81 (d, 3H
, J=10.7Hz), 4.41-4.44(m, I
H) *4.52 to 4.59 (m −2H),
5.67 (d, IH.

J=*1.4Hz), 6.17(d, IH, J-7,5
Hz), 8.03 (d.

IH,, J=7.5Hz) Example 5 2.3'-〇-Anhydro 1'-diethylphosphono-
β-D-Fructofuranosyluracil 2001 ffp of the compound of Example 1 was dissolved in 10 m of tetrahydrofuran, and under water cooling, 0.631 m of tetra-n
- A tetrahydro-7 run solution (1 molar solution) of butylammonium 70 lido was added and stirred for 15 minutes.

After adding 3 drops of water to the reaction solution and concentrating, the residue was subjected to silica gel column chromatography and eluted with methylene chloride/methanol (85:15 to 80:20) to give 79.711.
The title compound of 9C (yield: 65 J) was obtained.

Physical properties of the main product: Properties: Amorphous crystals [α]. --18,8° (methanol) UV, C vector (ethanol, λmax s nm): 247 (
g=7360), 225 ($=8910) x nsu-e
ri) Le (KBr t tyn): 1660
* 1540 * 148 ONMR Spect A, CD
MSO-d6. δppm): 1.14 (t*3H, J=
6.9Hz'), 1.22 (t, 3H, J=7.IH2
).

3.93 (q, 2H, J=6.9Hz), 4.00-4
．． 15 (m, 3H) -4,30~4.35 (m, IH)
, 4.55-4.65 (m, IH).

4.94 (t, IH, J = 5.2Hz), 5.28 (d
, IH, J=2.3Hz), 5.83(d,4H,J=
7.4Hz), 5.92(d, IH.

J=x4.6Hz), 6.82-6.95 (m, IH)
, 8.00 (d, IH.

J=7.4H1) Mass spectrum k (m/z)", 368 (M"
-24) Physical properties of the other isomer: [α)D--37,1° (methanol) Reference Example 1 2.3'-0-anhydro-4',6'-0-(1
, 1,3,3-tetraisofurovir disiloxane-1,
3-diyl)-β-D-fructofuranosyluracil 2
．． 3'-〇-Anhydro β-D-7lacto7lanosyluracil 9091f9 (3.55 mmol) was dissolved in 50-Da dimethylformamide and 990 Da imidazole (14.56 mmol) was added.

Cool this solution to -20°C and 1,3-dichloro-1,1
,3,3-tetrainofurovir disiloxane (1,29
1j, 3.75 mmol) was added. After stirring at -20°C for 30 minutes and at room temperature for 3 hours, the solvent was distilled off. The residue was diluted with chloroform, and the chloroform layer was washed with water and dried over anhydrous magnesium sulfate to obtain a crude product of 1,609-.

This crude product was subjected to silica column chromatography and eluted with methanol/methylene chloride (5:95) to obtain the title compound 928 da (yield 53%).

Properties: Colorless oily substance IR spectrum (film p"'-'): 32
50 t2950.2860,1630,1530,1
475,1405NMR spectrum # (CDC15, δ
ppm): 0.9-1.15 (m.

28H), 3.82-4.01 (m, 4H), 4.16
(dd, IH.

J=13.5, 6.0Hz), 4.47(dd, IH,
J = 5.2, 8.4 Hz), 5.41 (d, IH, J
=5.2Hz), 5.92(d, IH.

J = 8-I Hz) + 5.98 (d d
t I Ht J =9.2 t 6. (OHz).

7.33 (d, IH, J = 8.1 Hz) Reference example 2 2.3'-0-anhydro-1'-methoxy-4',6
1-〇-(tetraisofurovirdisiloxane-1,3-
Soil)-β-D-7lacto72nosyluracil Compound 2.12 of Reference Example 1? Dissolve 42-d in anhydrous dimethyl sulfoxide to obtain anhydrous benzene 42d.

0.3417 (1 equivalent) of anhydrous pyridine, 0.16 JE/ (0.5 equivalent) of trifluoroacetic acid, and 2.63 i (3 equivalent) of dicyclohexylcardiimide were sequentially added and allowed to stand overnight at room temperature.

The insoluble matter was separated from each other, ethyl acetate was added to the filtrate, the mixture was washed twice with water, dried (anhydrous magnesium sulfate), and concentrated.

The main product has the following physical properties.

IRx vector (KBr, m): 1660,154
0゜47O NMR spectral (CDCl2 pδppm): 0
．． 8-1.2 (m.

28H), 3.50 (a, 3H), 3.8-4.2 (
m, 4H), 4.3"'4.6 (m, IH), 5.06
(brs, IH), 5.33 (d, IH.

J = 6-OHz) 15.90 (d t I
H* J =7-8 Hz), 7-60 (d*
IH, J=7.8Hz) Physical property NMR spectrum k of the other isomer (CDC1,, δppm):
0.8~1.2 (m t28 H) t 3.50
(s t 3 H) -3,8~4.2 (m -4H
), 4.3"'4.6 (m, IH), 4.98 (br
a, IH), 5.43 (d, IH.

Jx6.0Hz), 5.94 (d, LH, J-7, 8H
$),? , 38 (d.

IH, J=7.5 Hz) [Effects of the Invention] The anhydrophosphononucleoside of the compound of the present invention has activity against murine leukemia P388. For example, the compound of Example 4 exhibits significant antitumor activity at a dose of 100 In9/kg. In view of this, the compound of the present invention is expected to be developed in the future as a new antitumor and antiviral drug.

Claims (4)

[Claims] (1) Anhydrophosphononucleoside derivatives having the general formula ▲ Numerical formulas, chemical formulas, tables, etc.▼ (in the formula, R^1 and R^2 are the same or different and represent a hydrogen atom or a lower alkyl group). (2) The compound according to claim 1, wherein the lower alkyl group is a methyl group, ethyl group, propyl group or butyl group. (3) General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (In the formula, R^3 represents a hydroxyl group protecting group, and R^4 represents a lower alkyl group). , chemical formulas, tables, etc. ▼ (In the formula, R represents a lower alkyl group) is reacted with a dialkyl phosphite in the presence of a base, followed by deprotection, or reacted in the presence of a base, deprotected, and further deprotected. Anhydrophosphonosine having the general formula ▲mathematical formula, chemical formula, table, etc.▼ (in the formula, R^1 and R^2 are the same or different and represent a hydrogen atom or a lower alkyl group) characterized by alkylation. Method for producing nucleoside derivatives. (4) The method according to claim 3, wherein the base is trialkylamine, sodium alkoxide, potassium alkoxide, or sodium hydride.