JP7373598B2 - Method for producing dinucleoside polyphosphate compounds - Google Patents

Description

The present invention relates to a method for producing a dinucleoside polyphosphoric acid compound, and more particularly, to a method for producing a dinucleoside polyphosphoric acid compound, a salt thereof, or a hydrate thereof from a nucleoside phosphoric acid compound in the presence of a metal halide.

Nucleosides are glycoside compounds in which a nucleobase and a pentose are linked through an N-glycosidic bond. When a phosphate group is attached to a nucleoside by a phosphorylating enzyme, it becomes a nucleotide, which is the basic component of a DNA chain. .

Dinucleoside polyphosphoric acid or its salt, which is a type of nucleotide, is stable in the body similar to substances in the body, and is known to be effective in treating diseases accordingly.

In particular, P ¹ , P ⁴ -di(uridine 5'-)tetraphosphate (hereinafter referred to as "UP ₄ U") represented by the following chemical formula 1a or a salt thereof may be used to treat xerophthalmia or decreased tear secretion. It is used as a therapeutic agent for corneal and conjunctival epithelial disorders associated with the disease, and is a compound that is expected to be developed as an expectorant or pneumonia treatment agent because it has a sputum excretion-inducing effect.

As a conventional method for synthesizing UP ₄ U, Non-Patent Document 1 describes the synthesis of uridine 5'-cyclic triphosphate and uridine produced by a dehydration condensation reaction of uridine 5'-triphosphate (hereinafter referred to as UTP). A conventional production method of reacting with 5'-monophosphoric acid (hereinafter referred to as UMP) has been reported, and Patent Document 1 reports on an improved production method.

Patent Document 2 discloses that uridine, UMP, UDP, or UTP and a salt thereof and a uridine nucleotide compound are dissolved in a polar, aprotic organic solvent and a hydrophobic amine, and a monophosphorylating agent or a diphosphorylating agent is used as a phosphorylating agent. A method for producing UP ₄ U using carbodiimide, active carbonyl, or active phosphorus as an activator has been reported.

Further, Patent Document 3 describes that uridine 5'-diphosphate (UDP), UMP or pyrophosphoric acid, and a group consisting of imidazole, benzimidazole or 1,2,4-triazole which may have a substituent. A phosphoric acid active compound synthesized by condensation with a selected compound and a phosphoric acid compound or a salt thereof selected from the group consisting of UMP, UDP, UTP and pyrophosphoric acid are combined in water in the presence of metal ions. Alternatively, a method for producing UP ₄ U by reaction in a hydrophilic organic solvent has been reported.

However, the conventional method for producing UP ₄ U requires a large number of complicated processes such as metal salt exchange reactions, resulting in a problem in that the synthesis efficiency and purity of the final compound are reduced.

Specifically, conventional methods for producing _UP4U such as Patent Document 2 and Patent Document 1 require that a metal salt of a phosphoric acid compound be reacted in the form of an amine salt such as tributylamine or triethylamine immediately before the reaction. No. As a result, the cumbersome process generally involves converting a metal salt, such as the sodium salt of a uridine phosphate compound, into a phosphate compound in the free acid form by ion-exchange resin column chromatography, followed by salt formation with an amine. is necessary. Performing such steps may result in a decrease in the synthesis efficiency and purity of the final compound. In particular, in the case of the above synthetic reaction using UTP, which is known to be a very unstable substance, the purity tends to decrease, and organic salts such as UTP amine salts, which are sensitive to moisture, have a high hygroscopicity. The problem is that it is very difficult to store and maintain quality.

In addition, the production method described in Patent Document 3 can synthesize UP ₄ U with a high yield of 45 to 94%, but the salt exchange step reaction for preparing the phosphoric acid compound organic salt mentioned above is This requires a corresponding dehydration process, and after synthesizing a phosphoric acid active compound in an organic solvent, the solvent must be removed by a vacuum concentration process, and the pH must be adjusted in the presence of water to avoid the hassle. Therefore, there is a problem that the purity of the compound is reduced.

Therefore, for the reasons mentioned above, it is desirable to produce dinucleoside polyphosphate compounds with high purity and high yield under environmentally friendly reaction conditions without any troublesome process steps, and to produce dinucleoside polyphosphate compounds suitable for industrial mass production. A manufacturing method is required.

International Publication No. 2008/012949 International Publication No. 1999/05155 International Publication No. 2014/103704

Bioorganic & Medicinal Chemistry Letters, 11, (2001), 157-160

An object of the present invention is to provide a method for producing dinucleoside polyphosphoric acid, its salts and hydrates, which is suitable and efficient for mass production, harmless to the environment, and capable of high purity and mass production. be.

The present invention provides a method for producing a dinucleoside polyphosphoric acid represented by the following chemical formula 1, a salt thereof, and a hydrate thereof.

前記化学式において、
Ｒ_１およびＲ_２は、互いに同一であるかまたは異なり、各々独立してピリミジン塩基であり；
Ｒ_３は、ピリミジン塩基であり；
ｎは、２～６の整数であり；
ｍは、１～３の整数である。 In one embodiment of the invention,
(S-1) a nucleoside phosphoric acid compound represented by the following chemical formula 2, a salt thereof, or a hydrate thereof;
Provided is a method for producing a dinucleoside polyphosphoric acid represented by the following chemical formula 1, a salt thereof, or a hydrate thereof, which includes a step of reacting a condensing agent of carbodiimides; and a metal ion in the presence of a solvent.

In the chemical formula,
R ₁ and R ₂ are the same or different from each other and are each independently a pyrimidine base;
R ₃ is a pyrimidine base;
n is an integer from 2 to 6;
m is an integer from 1 to 3.

In the present invention, the pyrimidine base can be selected from the group consisting of cytosine, uracil, or thymine.

In one embodiment of the invention said base is uracil.

In one embodiment of the invention, said n is 4.

In one embodiment of the present invention, the nucleoside polyphosphoric acid represented by the chemical formula 1 is represented by the following chemical formula 1a.

According to the production method of the present invention, by using the chemical formula 2, its salt, or its hydrate, which can be purchased in the market, without a separate conversion process, the equipment required for production can be reduced, and the equipment used in the conversion process can be reduced. time, effort, and cost can be saved, and the generation of impurities can be correspondingly minimized.

In one embodiment of the present invention, the nucleoside phosphoric acid compound represented by Chemical Formula 2 is represented by any one of the following Chemical Formulas 2a to 2c.

In one embodiment of the invention, the salt of the nucleoside phosphoric acid compound is a metal salt or an amine salt.

The metal salt of the nucleoside phosphate compound may be selected from the group consisting of lithium, sodium, potassium, magnesium, calcium, barium, cerium, iron, nickel, copper, zinc and boron, preferably lithium, sodium or Alkali metal salts of potassium and alkaline earth metal salts of calcium or magnesium may also be used.

In one embodiment, the amine salt may be a tertiary amine, specifically an alkyl amine such as trimethylamine, triethylamine, tributylamine, tripentylamine, trihexylamine, triethanolamine, or pyridine. The chain may be selected from the group consisting of C ₁ -C ₆ trialkylamines and cyclic trialkylamines.

In the present invention, a condensing agent refers to a compound added as a reaction aid in a condensation reaction.

前記化学式３において、
Ｒ_４およびＲ_５は、互いに同一であるかまたは異なり、各々独立して炭素数１～６の直鎖、分枝鎖もしくは環状鎖のアルキル基であり、前記アルキル基はアルキルアミン基で選択的に置換されてもよい。 In one embodiment of the present invention, the carbodiimide condensing agent is represented by Formula 3 below.

In the chemical formula 3,
R ₄ and R ₅ are the same or different and each independently represents a linear, branched or cyclic alkyl group having 1 to 6 carbon atoms, and the alkyl group is selectively an alkylamine group. may be replaced with

Specifically, R ₄ and R ₅ are the same or different and each independently represents ethyl, isopropyl, cyclohexyl or dimethylaminopropyl.

The condensing agents for the carbimides include 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) or a salt thereof, N,N'-diisopropylcarbodiimide (DIC), and N,N'-dicyclohexylcarbodiimide (DCC). selected from the group consisting of.

In another embodiment of the present invention, step (S-1) may further include a triazole additive. The triazole additive may specifically be hydroxybenzotriazole (HOBt) or 1-hydroxy-7-azabenzotriazole (HOAt). The triazole additive has the effect of increasing the efficiency of the carbodiimide condensing agent.

In one embodiment of the invention, said metal ions are derived from the group consisting of metal chlorides, bromides, nitoxides, sulfates and acetates.

前記化学式４において、
ａは、Ｍのモル数であり、
ｂは、Ｍのイオン価数（Ｉｏｎｉｃ ｖａｌｅｎｃｙ）であり、
Ｍは、カルシウム、マグネシウム、セリウム、鉄、リチウム、アルミニウム、チタニウムまたはナトリウムであり、
ｃは、Ｘのモル数であり、
ｄは、Ｘのイオン価数であり、
Ｘは、ハロゲン、カーボネート、アセテート、ニトレート、トリフレート、サルフェート、カルボキシレートまたはこれらの誘導体であり、
ａとｂの積の値と、ｃとｄの積の値とは同一である。 In one embodiment of the invention, the metal ion may be an ion of a metal selected from the group consisting of calcium, magnesium, cerium, iron, lithium, aluminum, titanium, and sodium.
In one embodiment of the present invention, the metal ion may be derived from a metal salt catalyst represented by Formula 4 below.

In the chemical formula 4,
a is the number of moles of M,
b is the ionic valency of M,
M is calcium, magnesium, cerium, iron, lithium, aluminum, titanium or sodium;
c is the number of moles of X,
d is the ionic valence of X,
X is a halogen, carbonate, acetate, nitrate, triflate, sulfate, carboxylate or a derivative thereof,
The value of the product of a and b is the same as the value of the product of c and d.

In one embodiment of the invention, the metal ions include calcium salts such as calcium chloride, calcium acetate, calcium carbonate, calcium sulfate, calcium phosphate, magnesium salts such as magnesium chloride, magnesium carbonate, magnesium sulfate, magnesium bromide, cerium chloride, etc. , metal ions derived from cerium salts such as cerium fluoride, cerium iodide, cerium nitrate, and cerium trifluoroacetate, specifically, calcium chloride, cerium chloride, lithium iodide, magnesium chloride, and iron chloride. The metal ion may be derived from, but is not limited to, and may include all forms of anhydride or hydrate.

In the production method according to the present invention, by using catalysts that are inexpensive and harmless to the human body, it is possible to dramatically shorten the reaction time and provide a high yield of dinucleoside polyphosphoric acid. It can be applied very suitably to

In one embodiment of the present invention, the solvent is water, an organic solvent, or a mixed solvent of water and an organic solvent.

In one embodiment of the present invention, water can be used alone as the solvent. Since the production method according to the present invention can use water alone, when it is applied to industrial mass production under environment-friendly reaction conditions, there are problems of environmental pollution, costs incurred due to the use of organic solvents, and waste organic matter. It is economical and environmentally friendly because the cost of processing the solvent can be minimized.

In one embodiment, the solvent may be a mixed solvent of water and an organic solvent.

In yet another embodiment, the organic solvent is selected from the group consisting of C ₁ -C ₈ alcohols, C ₃ -C ₁₀ ketones, 1,4-dioxane, acetonitrile, N,N-dimethylformamide, and dimethyl sulfoxide. be done.

In one embodiment of the present invention, in step (S-1), the equivalent of the condensing agent and the equivalent of the metal ion are the same or different, each of which is 1 mol of the nucleoside phosphoric acid compound represented by the chemical formula 2. The reaction is carried out in an amount of 0.1 to 30.0 molar equivalents. In particular, the equivalent of the condensing agent and the equivalent of the metal ion are the same or different, and each is 1.0 to 10.0 molar equivalent per 1 molar equivalent of the nucleoside phosphoric acid compound represented by the chemical formula 2. react.

In one embodiment of the present invention, the reaction temperature in step (S-1) can be reasonably carried out within a reaction temperature range that allows the synthesis of general pharmaceuticals, and specifically, The reaction temperature in step 1) may be 0°C to 50°C, more specifically, the reaction is carried out at 5°C to 35°C.

In one embodiment of the present invention, the method further includes the step of (S-2) solidifying and separating and purifying the product obtained from step (S-1).

The step (S-2) is a step in which the reaction solution obtained in the step (S-1) is solidified and then purified using various methods so that it can be used as a medicine.

Specifically, the step (S-2) is a first step of generating a crude solid by injecting or back-injecting an organic solvent after completing the step (S-1) and filtering it. including.

In one embodiment of the present invention, the organic solvent in step (S-2) may be a hydrophilic organic solvent. The hydrophilic organic solvent is selected from the group consisting of C ₁ -C ₈ alcohols, C ₃ -C ₁₀ ketones, 1,4-dioxane, acetonitrile, N,N-dimethylformamide and dimethylsulfoxide. Specifically, it can be selected from the group consisting of methanol, ethanol, propanol, isopropanol, etc., acetone, 1,4-dioxane, N,N-dimethylformamide, and dimethyl sulfoxide.

In one embodiment of the present invention, the step (S-2) includes dissolving the generated crude solid in water after the first step, adsorbing it to an anion exchange resin and eluting the target compound, and then removing sodium. It may further include a second step of converting the form to obtain the final compound.

In the present invention, the anion exchange resin includes weakly basic anion exchange resins (Amberlite IRA67, Diaion SA-11A, etc.), strong basic anion exchange resins (Amberlite IRA402, Diaion PA-312, etc.). ), a weakly acidic cation exchange resin (such as Diaion WK-30) or a strongly acidic cation exchange resin (such as Diaion PK-216, DOWEX 50WX2) can be used. In particular, the target compound can be eluted with higher purity through the weakly basic anion exchange resin.

In one embodiment of the present invention, a nucleoside phosphoric acid compound represented by the chemical formula 2, a salt thereof, or a hydrate thereof; a condensing agent represented by the chemical formula 3; and a metal salt represented by the chemical formula 4. P ¹ , P ⁴ -di(uridine 5'-)tetraphosphoric acid represented by the chemical formula 1a, a salt thereof, or a hydrate thereof, including a step of reacting with a metal ion derived from a catalyst in the presence of a solvent. A manufacturing method can be provided.

What has been mentioned above may equally apply insofar as they do not contradict each other.

As in the production method according to the present invention, the nucleoside phosphoric acid compound represented by the chemical formula 2 is reacted with water, which is an environmentally friendly solvent, together with a condensing agent and metal ions, resulting in a relatively simple process. Accordingly, the dinucleoside polyphosphoric acid represented by the chemical formula 1, its salt, or its hydrate can be produced with high efficiency and high purity.

According to the present invention, dinucleoside polyphosphoric acid can be converted into dinucleoside polyphosphoric acid at a high reaction conversion rate without a complicated and troublesome conversion step of converting a commercially available form of a nucleoside phosphoric acid compound or its metal salt into a free acid or organic salt form of the nucleoside phosphoric acid compound. can be synthesized. As a result, since almost no side reaction products are produced, purification is very easy, and a high-quality compound for use as a raw material for pharmaceuticals can be obtained. In addition, by using water as a solvent alone or in combination with an organic solvent, it is a very efficient and environmentally friendly synthesis method that can minimize environmental pollution during industrial mass production. be. Furthermore, it is widely applicable to the synthesis of substituted phosphoric acid compound derivatives with various structures.

These are the results of nuclear magnetic resonance analysis ( ¹ H NMR) of UP ₄ U (diquafosol) synthesized in Example 1. 1 is HPLC data of primary UP ₄ U (diquafosol) obtained after completion of the reaction in Example 1. 1 is HPLC data of UP ₄ U (diquafosol) finally obtained after synthesis and purification in Example 1.

Examples are presented below to aid in understanding the invention. However, the following examples are merely provided for easier understanding of the present invention, and the content of the present invention is not limited by the following examples.

In addition, the reagents and solvents mentioned below were purchased from Chinese raw material manufacturers Sigma-Aldrich Korea and TCI, HPLC was measured using Agilent Technologies' 1200 Series, and ¹ H NMR was: Measurements were made using a Bruker 400 ultraShield NMR Spectrometer. Purity was determined by HPLC area %.

The HPLC conditions used in the present invention were as follows, and the purity of _UP4U (diquafosol) was measured after the reaction or in the reaction mixture.
Detector: Ultraviolet absorption photometer (measurement wavelength: 260 nm)
Column: YMC-Pack ODS-AQ (4.6mm x 250mm, 5μm)
Mobile phase: 0.4% potassium dihydrogen phosphate aqueous solution Flow rate: 0.5 mL/min
Sample: _UP4U (diquafosol) 10mg/mobile phase 10mL
Injection volume: 10μL

Example 1. Synthesis of UP4U _from UDP・2Na UDP・2Na salt (1000 g, 2.23 mol) was dissolved in 3.0 L of purified water and stirred at 10°C, then EDC・HCl (428 g, 2.23 mol) and CaCl ₂ - 2H ₂ O (328 g, 2.23 mmol) was sequentially added dropwise and stirred at 10° C. for 4 hours and 30 minutes. The reaction was monitored by HPLC.

After the reaction was completed, the obtained compound was solidified with 3.0 L of purified water and 12.0 L of EtOH, and stirred at room temperature for about 1 hour. The produced solid was filtered and dried to obtain 1,038 g (purity 94.1%) of the target UP ₄ U compound.

The obtained solid was dissolved in deionized water and adsorbed on a weakly basic anion exchange resin (Amberlite IRA series), and then sequentially eluted with deionized water, a low concentration hydrochloric acid solution, and a sodium chloride solution, and concentrated under reduced pressure. Afterwards, a solid was precipitated with ethanol. The solid was filtered and dried to obtain the target UP ₄ U·4Na (780 g, yield 80%, purity 99.95%).

Experimental example 1. Synthesis of _UP4U from UDP・2Na: Effect on metal salts UDP・2Na salt (500 mg, 1.12 mmol) was dissolved in 2 mL of purified water, and DIC (259 uL, 1.67 mmol) and various metal salts (1.67 mmol) were dissolved in 2 mL of purified water. ) were added dropwise in order and allowed to react at room temperature.

The reaction solution was analyzed by HPLC to determine the conversion rate to the target UP ₄ U and the purity of UP ₄ U in the reaction solution in comparison to UDP.2Na.

From the results in Table 1 above, in the synthesis of UP ₄ U from UDP 2Na salt, the conversion rate is 2% in the absence of metal salts, while it is 79% in the case of magnesium salts and 25% in the case of iron salts. % conversion rate, and when calcium was selected as the metal salt, a high conversion rate of 97% to 98% was shown, and the purity of the reaction solution was 83% to 84%, confirming a remarkable increasing effect.

That is, from the above results, when the nucleoside phosphoric acid compound represented by the chemical formula 2 of the present invention, its salt or its hydrate; the condensing agent; and the metal ion are reacted, the reaction rate is higher than when there is no metal ion. It was confirmed that UP ₄ U could be obtained in high yield and purity.

Experimental example 2. Synthesis of _UP4U from UDP・2Na: Effect on condensing agent UDP・2Na salt (500 mg, 1.12 mmol) was dissolved in 2 mL of purified water, and various condensing agents (1.67 mmol) and metal salt CaCl ₂・2H ₂ O (247 mg, 1.67 mmol) was sequentially added dropwise and reacted at room temperature.

The reaction solution was analyzed by HPLC to determine the conversion rate to the target UP ₄ U and the purity of UP ₄ U compared to UDP.

From the results in Table 2 above, when DIC, EDC/HCl, or DIC and HOBt is used as a condensing agent in the synthesis of UP ₄ U from UDP/2Na salt, the conversion rate to the target UP ₄ U is 98% or more. It was confirmed that the purity of UP ₄ U was 84% or more. In particular, it was confirmed that when EDC/HCl was used as a condensing agent, the reaction time was significantly reduced, the conversion rate to UP ₄ U was high, and the target compound UP ₄ U could be synthesized with high purity.

Experimental example 3. Synthesis of _UP4U from UDP・2Na: Effect of reaction temperature, equivalents of condensing agent and metal salt UDP・2Na salt (500 mg, 1.12 mmol) was dissolved in 2 mL of purified water, and the corresponding equivalents of condensing agent and metal were dissolved. Salt CaCl ₂ .2H ₂ O was added dropwise in order to react.

The reaction solution was analyzed by HPLC to determine the conversion rate to the target UP ₄ U and the purity of UP ₄ U compared to UDP.

From the results in Table 3 above, in the synthesis of UP ₄ U from UDP 2Na salt, at a high reaction temperature of 40°C or higher, the starting material and target compound are decomposed and the purity in the reaction solution is 45% to 72%. It got lower.

Experimental example 4. Synthesis of _UP4U from UDP/organic salt: Effect on metal salts UDP/2TBA salt (1 g, 1.30 mmol) was dissolved in 10 mL of DMF, and DIC (240 uL, 1.55 mmol) and various metal salts (1.55 mmol) were dissolved. ) were added dropwise in order and allowed to react at room temperature. The reaction solution was analyzed by HPLC to determine the conversion rate and purity of the target UP ₄ U compared to UDP.

UDP·2TEA salt (500 mg, 0.82 mmol) was dissolved in 5 mL of DMF, and DIC and various metal salts were added dropwise in the equivalent amounts shown below and reacted at room temperature. The reaction solution was analyzed by HPLC to determine the conversion rate and purity of the target UP ₄ U compared to UDP.

From the results in Table 4 above, in the synthesis of UP ₄ U from UDP organic salts (TBA salts or TEA salts), a high conversion rate of 83% to 99% can be achieved in the presence of calcium, iron, magnesium, lithium, or cerium salts. confirmed.

It was confirmed that UP ₄ U could be obtained with a high purity of 71% to 83% when the reaction was carried out in the presence of a calcium salt or a magnesium salt in the high conversion reaction solution.

That is, from the above results, when the nucleoside phosphoric acid compound represented by the chemical formula 2 of the present invention, a salt thereof or a hydrate thereof; a condensing agent; and a metal ion are reacted, high yield and high purity UP ₄ U can be obtained. It was confirmed that it is possible to obtain

Experimental example 5. Synthesis of UP ₄ U from cUTP: Effect on metal salts Dissolve UTP/TBA salt (5 g, 9.10 mmol) in 50 mL of DMF, make a cUTP solution with DIC (1.69 mL, 10.92 mmol), and quantify by HPLC. 1 ml (0.11 mmol) of the cUTP solution was taken, and a UMP/TBA salt solution (0.13 mmol) and various metal salts (0.132 mmol) were added dropwise in this order to react at room temperature.

The reaction solution was analyzed by HPLC to determine the conversion rate to the target UP ₄ U and the purity of UP ₄ U compared to UTP.

From the results in Table 5 above, in the synthesis of UP ₄ U from UTP/TBA salt, a high conversion rate of 83% to 99% was shown in the presence of calcium, iron, magnesium, lithium, or cerium salt; It can be confirmed that the conversion rate without it is significantly low at 41%.

From the above results, when the nucleoside phosphoric acid compound represented by the chemical formula 2 of the present invention, its salt or its hydrate; the condensing agent; and metal ions are reacted, the yield is higher than when there is no metal ion. It was confirmed that UP ₄ U of high purity could be obtained.

Comparative example 1. Synthesis of UP ₄ U from UDP: International Publication No. 2014/103704 UDP 2Na salt (50 g, 0.112 mmol) was dissolved in 400 mL of purified water and adsorbed on Dowex 50w x 2-100 strongly acidic cation exchange resin. After that, 1200 mL of purified water is passed through at a rate of 30 mL/min to elute the UDP solution from which Na salts have been removed. Inject tributylamine (80 mL, 0.336 mmol) into the eluate to neutralize it to pH 7 or higher, concentrate under reduced pressure at 60°C, and then azeotropically concentrate under reduced pressure with 1,4-dioxane several times to remove water. . The resulting residue was vacuum dried at room temperature for 12 hours to obtain 75 g of UDP/2TBA salt (yield: 87%, moisture: 1.2%).

UDP-2TBA (14.0 g, 18.1 mmol), in which Na salt was replaced with TBA salt in the above step, was dissolved in 46 mL of propionitrile, carbonyldiimidazole (8.8 g, 54.3 mmol) was added, and the mixture was heated at room temperature. The mixture was stirred for about 30 minutes and concentrated under reduced pressure to remove the solvent. After pouring 7 mL of purified water into the obtained residue and cooling it to about 5° C., UDP·2Na (4.1 g, 9.1 mmol) was added. After titrating the reaction solution to pH 3.9 using a 6N aqueous hydrochloric acid solution, a 60% aqueous ferric chloride solution (75 μL, 0.36 mmol) was added, and the mixture was stirred at 10° C. for 27 hours. The reaction solution was titrated to pH 10 with a 7.5N aqueous sodium hydroxide solution, and stirred at 5°C for 1 hour. At the same temperature, 90 mL of ethanol was injected, and the mixture was allowed to stand at the same temperature for 12 hours and filtered to obtain 13.8 g of UP ₄ U compound (purity: 86.9%) in solid form.

Based on the results of Experimental Examples 1 to 5, as in the production method according to the present invention confirmed in Example 1, a nucleoside phosphoric acid compound represented by the chemical formula 2, a salt thereof, or a hydrate thereof; a condensing agent ; and metal ions, it was confirmed that dinucleoside polyphosphoric acid can be synthesized in high yield and purity at a high reaction conversion rate without any troublesome conversion steps. Even when compared with Comparative Example 1, which is the most effective experimental method among the prior art, this method requires separate processing of the starting material in the form of a metal salt, such as a commercially available sodium salt, without the troublesome UDP salt replacement process. It was confirmed that the work time was dramatically improved due to the extremely simplified reaction process.

From the above results, it can be confirmed that the method for producing dinucleoside polyphosphoric acid, a salt thereof, or a hydrate thereof according to the present invention is industrially compatible and can be usefully applied to mass production.

Claims (17)

(S-1) a sodium salt of a nucleoside phosphoric acid compound represented by the following chemical formula 2;
a step of reacting a condensing agent of carbodiimides; and a metal ion; in the presence of a solvent; and (S-2) (i) obtaining a solid crude product after completing step (S-1); (ii ) The resulting crude product solid is dissolved in water, the resulting solution is adsorbed on an anion exchange resin, and the dinucleoside polyphosphoric acid represented by the following chemical formula 1 is eluted, and the dinucleoside polyphosphoric acid represented by the following chemical formula 1 is eluted. A method for producing a sodium salt of a dinucleoside polyphosphoric acid or a hydrate thereof represented by the following chemical formula 1, comprising a step of converting the dinucleoside polyphosphoric acid into a sodium form, the method comprising:
A method, wherein the metal ion is an ion of calcium:
In the chemical formula,
R ₁ and R ₂ are the same or different from each other and are each independently a pyrimidine base;
R ₃ is a pyrimidine base;
n is an integer of 4;
m is an integer from 1 to 3. 2. The method of claim 1, wherein the pyrimidine base is uracil. The method according to claim 1, wherein the nucleoside polyphosphoric acid represented by Chemical Formula 1 is represented by the following Chemical Formula 1a.
The method according to claim 1, wherein the nucleoside phosphoric acid compound represented by Chemical Formula 2 is represented by any one of the following Chemical Formulas 2a to 2c:
The method according to claim 1, wherein the carbodiimide condensing agent is represented by the following chemical formula 3:
In the chemical formula 3,
R ₄ and R ₅ are the same or different and each independently represents a linear, branched or cyclic alkyl group having 1 to 6 carbon atoms, and the alkyl group is selectively an alkylamine group. may be replaced with 6. The method of claim 5, wherein _R4 and _R5 are the same or different and are each independently ethyl, isopropyl, cyclohexyl or dimethylaminopropyl. The condensing agent for the carbodiimides is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) or a salt thereof, N,N'-diisopropylcarbodiimide (DIC), and N,N'-dicyclohexylcarbodiimide (DCC). 2. The method of claim 1, wherein the method is selected from the group consisting of: 2. The method of claim 1, wherein the metal ions are from the group consisting of metal chlorides, bromides, nitoxides, sulfates, and acetates. The method according to claim 1, wherein the metal ion is derived from a metal salt catalyst represented by the following chemical formula 4:
In the chemical formula 4,
a is the number of moles of M,
b is the ionic valence of M,
M is calcium;
c is the number of moles of X,
d is the ionic valence of X,
X is a halogen, carbonate, acetate, nitrate, triflate, sulfate, carboxylate or a derivative thereof,
The value of the product of a and b is the same as the value of the product of c and d. The method according to claim 1, wherein the solvent in step (S-1) is water or a mixed solvent of water and an organic solvent. 11. The organic solvent according to claim 10, wherein the organic solvent is selected from the group consisting of C ₁ -C ₈ alcohols, C ₃ -C ₁₀ ketones, 1,4-dioxane, acetonitrile, N,N-dimethylformamide and dimethyl sulfoxide. Method described. The equivalent of the condensing agent and the equivalent of the metal ion are the same or different, and each reacts in an amount of 0.1 mol to 30.0 mol equivalent per 1 mol equivalent of the nucleoside phosphoric acid compound represented by the chemical formula 2. 2. The method according to claim 1. The method according to claim 1, wherein the reaction temperature is 0°C to 50°C. 2. The method of claim 1, wherein the step of obtaining a crude product solid is producing a crude product solid by injecting or back-injecting an organic solvent and filtering the crude product solid. 15. The method according to claim 14 , wherein the organic solvent is a hydrophilic organic solvent. 15. The hydrophilic organic solvent is selected from the group consisting of C ₁ -C ₈ alcohols, C ₃ -C ₁₀ ketones, 1,4-dioxane, acetonitrile, N,N-dimethylformamide and dimethyl sulfoxide. The method described in. 2. The method of claim 1, wherein the anion exchange resin is a weakly basic anion exchange resin or a strongly basic anion exchange resin.

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