Method for producing ribose phosphate compound and method for preventing solute deposition in supersaturated reaction solution

Abstract

Provided is a method for producing a ribose phosphate compound, the method comprising hydrolyzing a compound represented by general formula (3) in the presence of an acidic compound by a pressurized flow reaction, thereby producing a ribose phosphate compound represented by general formula (1). R1 represents a hydrogen atom, a hydroxy group, or OR11, and R11 represents a group represented by general formula (2). R2 and R3 each represent a hydrogen atom or a group represented by general formula (2). The two R12 each represent a hydrogen atom or an alkali metal ion. * indicates a bonding part with an oxygen atom. R5 represents an adenyl group, a guanyl group, a cytosyl group, a uracil group, or a thyminyl group.　The compounds represented by general formulae (1) and (3) each have at least one group represented by general formula (2) per molecule.

Description

Method for producing ribose phosphate compound and method for preventing solute precipitation in supersaturated reaction solution

[0001] The present invention relates to a method for producing a ribose phosphate compound and a method for preventing solute precipitation in a supersaturated reaction solution.

[0002] Ribose phosphate compounds, such as ribose 5-phosphate and deoxyribose 5-phosphate, are used as raw materials for antibiotics, antiviral drugs, antisense medicines, etc. Ribose phosphate compounds are also used as substrates (synthetic raw materials) for nucleic acid synthesis.

[0003] It is known that ribose phosphate compounds can be obtained by hydrolyzing nucleotides using a solid acid catalyst or hydrochloric acid as a catalyst (Non-Patent Documents 1 and 2). Another technique for hydrolyzing nucleotides is the hydrolysis of pyrimidine nucleotides using sulfuric acid as a catalyst (Patent Document 1). Both of these hydrolysis reactions are carried out in a batch system.

[0004] Chem. Commun., 2017, Vol. 53, pp. 4919-4921 Bioorganic & Medicinal Chemistry Letters, 2021, Vol. 44, pp. 128105

[0005] Special Publication No. 48-30637

[0006] The hydrolysis of nucleotides in the presence of a catalyst is more efficient at high temperatures. Because this hydrolysis reaction is carried out using an aqueous solvent (water or a solvent mainly composed of water), the reaction temperature can be set near the boiling point of water (approximately 100°C). Nucleotides can be sufficiently hydrolyzed by a hydrolysis reaction at approximately 100°C for several hours. However, the ribose phosphate compounds produced by hydrolysis are heat-labile, and if nucleotide hydrolysis is carried out at approximately 100°C for several hours, much of the ribose phosphate compounds produced will be thermally decomposed. In other words, in the nucleotide hydrolysis reactions described in the above-mentioned Non-Patent Documents 1 and 2 and Patent Document 1, there is a so-called trade-off between sufficient hydrolysis and suppression of decomposition of the ribose phosphate compounds produced, which limits the improvement in the yield of ribose phosphate compounds.

[0007] An object of the present invention is to provide a method for producing a ribose phosphate compound, which enables the compound to be obtained in higher yield from a nucleotide or an analogue thereof by hydrolysis reaction.

[0008]

[0005] In view of the above problems, the present inventors have conducted extensive research and found that, when obtaining ribose phosphate compounds by the hydrolysis of nucleotides, applying a pressurized flow reaction using an acidic compound as a catalyst increases the boiling point of the solvent, enabling a liquid-phase reaction at a higher temperature, thereby enabling sufficient hydrolysis of nucleotides in a reaction time significantly shorter than conventional methods, thereby achieving both a shortened reaction time and an improved yield of ribose phosphate compounds at a high level. The present invention was completed based on these findings and through further research.

[0009] The above-mentioned object of the present invention is achieved by the following means: [1] A method for producing a ribose phosphate compound, which comprises subjecting a compound represented by the following general formula (3) to a hydrolysis reaction in the presence of an acidic compound by a pressurized flow reaction to produce a ribose phosphate compound represented by the following general formula (1): In general formula (1), R １ is a hydrogen atom, a hydroxy group, or OR １１ indicates R １１ represents a group represented by the following general formula (2): ２ and R ３ each independently represents a hydrogen atom or a group represented by the following general formula (2): In the general formula (2), two R １２ Each independently represents a hydrogen atom or an alkali metal ion. * represents a link to an oxygen atom. In general formula (3), R １ , R ２ , R ３ and R １１ are R in the above general formula (1), respectively. １ , R ２ , R ３ and R １１ It is synonymous with R ５represents an adenyl group, a guanyl group, a cytosyl group, a uracil group, or a thyminyl group. However, the compounds represented by the general formulas (1) and (3) each have at least one group represented by the general formula (2) in the molecule. [2] The method for producing a ribose phosphate compound according to [1], wherein the solvent for the pressure flow reaction contains water. [3] The method for producing a ribose phosphate compound according to [1] or [2], wherein the acidic compound has a pH of 2.0 or less when made into an aqueous solution at a concentration of 1 mol / L. [4] The method for producing a ribose phosphate compound according to any of [1] to [3], wherein the acidic compound has a sulfo group. [5] The method for producing a ribose phosphate compound according to any of [1] to [4], wherein the pressure flow reaction comprises heating a solution containing the compound represented by the general formula (3) flowing through a flow channel to cause the hydrolysis reaction, cooling the reaction solution in which the hydrolysis reaction has occurred, and then warming the cooled reaction solution. [6] The method for producing a ribose phosphate compound according to [5], wherein the temperature of the solution is increased to a range of 105 to 160°C by heating, the temperature of the reaction solution is cooled to a range of 0 to 40°C by cooling, and the temperature of the reaction solution is increased to a range of 10 to 60°C by warming. [7] The method for producing a ribose phosphate compound according to [5] or [6], wherein the time for subjecting the solution to the heating is 30 to 300 seconds and the time for subjecting the solution to the cooling is 1 to 30 seconds. [8] The method for producing a ribose phosphate compound according to any of [1] to [7], wherein the pressure of the pressurized flow reaction is 0.3 to 20.0 MPa. [9] The method for producing a ribose phosphate compound according to any of [1] to [8], wherein the pressure of the pressurized flow reaction is controlled by a back pressure valve.

[10] １ is a hydrogen atom or a hydroxy group, ２ is a hydrogen atom, and the above R ３is a group represented by the general formula (2).

[11] A method for preventing solute precipitation in a supersaturated reaction solution, comprising, when a supersaturated reaction solution flowing through a channel of a pressurized flow-type reaction channel passes through an outlet of the pressurized flow-type reaction channel and is released from pressure, adjusting the temperature of the supersaturated reaction solution upstream of the outlet to prevent solute precipitation downstream of the outlet.

[12] The method for preventing solute precipitation in a supersaturated reaction solution according to

[11] , wherein the pressure of the pressurized flow-type reaction channel is controlled by a back-pressure valve.

[13] The method for preventing solute precipitation in a supersaturated reaction solution according to

[11] or

[12] , wherein the temperature adjustment is performed by raising the temperature of the supersaturated reaction solution to a range of 10 to 60°C.

[14] The method for preventing solute precipitation in a supersaturated reaction solution according to any one of

[11] to

[13] , wherein the temperature of the supersaturated reaction solution before the temperature adjustment is in the range of 0 to 40°C.

[0010] In the present invention, a numerical range expressed using "to" means a range that includes the numerical values ​​before and after "to" as the lower and upper limits.

[0011] According to the method for producing a ribose phosphate compound of the present invention, it is possible to obtain a ribose phosphate compound in a higher yield from a nucleotide or an analog thereof by a hydrolysis reaction.

[0012] 1 is a diagram showing one embodiment of a flow reaction system suitable for carrying out the method for producing a ribose phosphate compound of the present invention.

[0013] [Method for Producing Ribose Phosphate Compounds] The method for producing a ribose phosphate compound of the present invention (hereinafter referred to as the "production method of the present invention") is a method for obtaining a ribose phosphate compound by hydrolyzing a nucleotide or an analog thereof in the presence of an acidic compound through a pressurized flow reaction. The ribose phosphate compound obtained by the production method of the present invention may be neutralized after the hydrolysis to form a salt.

[0014] <Nucleotide or Analog Thereof> In the present invention, a "nucleotide or analog thereof" is a reaction raw material that causes a hydrolysis reaction, and is a compound represented by the following general formula (3): In the present invention, the compound represented by general formula (3) represents the state before being mixed with an acidic compound.

[0015]

[0016] In general formula (3), R １ is a hydrogen atom, a hydroxy group, or OR １１ indicates R １１ represents a group represented by the following general formula (2): ２ and R ３ R each independently represents a hydrogen atom or a group represented by the following general formula (2): ５ represents an adenyl group, a guanyl group, a cytosyl group, a uracil group, or a thyminyl group.

[0017]

[0018] In the general formula (2), two R １２ Each independently represents a hydrogen atom or an alkali metal ion. * represents a link to an oxygen atom. R １２ Examples of alkali metal ions that can be used as R include lithium ions, sodium ions, potassium ions, and cesium ions. １２ is preferably a hydrogen atom, a sodium ion or a potassium ion, more preferably a hydrogen atom.

[0019] However, the compound represented by the general formula (3) has at least one group represented by the general formula (2) in the molecule. ３ is preferably a group represented by the general formula (2). The compound represented by the general formula (3) preferably has one or two groups represented by the general formula (2) in the molecule, and more preferably has one group. The compound represented by the general formula (3) is more preferably a compound represented by the general formula (3) ３ is a group represented by the general formula (2), and R １ is a hydrogen atom or a hydroxy group, and R ２ is a hydrogen atom. In this case, R １is a hydrogen atom, the compound represented by the above general formula (3) is a deoxyribonucleotide, and R １ is a hydroxy group, the compound represented by the above general formula (3) is a ribonucleotide.

[0020] <Acidic Compound> In the present invention, an "acidic compound" acts as a reaction catalyst (acid catalyst) for a hydrolysis reaction. In the present invention, an "acidic compound" means a compound that, when prepared as an aqueous solution with a concentration of 1 mol / L (1 M) (a solution in which the compound is dissolved in water to a concentration of 1 mol / L), causes the pH of this aqueous solution to be 5.0 or less. This pH is measured at 25°C. The pH of the aqueous solution is preferably 4.0 or less, more preferably 3.0 or less, even more preferably 2.5 or less, even more preferably 2.0 or less, and even more preferably 1.5 or less. The pH of the aqueous solution is usually 0.0 or more.

[0021] A preferred example of the acidic compound is a sulfo group (—SO ３ Among them, sulfuric acid (H ２ SO ４ ) is suitable as the acidic compound.

[0022] <Pressurized Flow Reaction> In the present invention, a "pressurized flow reaction" refers to a reaction in which a flow channel is pressurized and a liquid containing reaction raw materials is circulated through the pressurized flow channel. Furthermore, a "pressurized flow reaction channel" refers to a reaction channel portion within a reaction system in which a pressurized flow reaction is carried out, the portion of the reaction channel being pressurized. For example, when the pressure in the reaction channel in a pressurized flow reaction is adjusted using a back-pressure valve located downstream of the reaction channel, the back-pressure valve and its upstream and downstream sides together constitute the pressurized flow reaction system, and the pressurized flow reaction channel extends from the channel inlet (inlet) through which a liquid is introduced into the pressurized reaction channel to the back-pressure valve (excluding the portion downstream of the back-pressure valve). Therefore, the back-pressure valve corresponds to the "outlet" of the pressurized flow reaction channel. In the following description, the pressurized flow reaction used in the production method of the present invention will also be referred to as the "pressurized flow reaction of the present invention." Furthermore, the "pressurized flow reaction channel" used in the "pressurized flow reaction of the present invention" will also be referred to as the "pressurized flow reaction channel of the present invention."

[0023] "Pressurizing" in the pressurized flow reaction of the present invention means that the pressure in the pressurized flow reaction channel is 0.2 MPa or higher. By heating the liquid in the channel under such pressurized conditions, the aqueous solvent can be heated to a temperature above 100°C, thereby further increasing the efficiency of the hydrolysis reaction. The pressure in the pressurized flow reaction channel is more preferably 0.3 to 20.0 MPa, even more preferably 0.4 to 15.0 MPa, even more preferably 0.5 to 10.0 MPa, even more preferably 0.6 to 8.0 MPa, even more preferably 0.8 to 7.0 MPa, and even more preferably 1.0 to 5.0 MPa.

[0024] In the pressurized flow reaction of the present invention, the manner in which the reactant solution containing the compound represented by the general formula (3) flows through the pressurized flow reaction channel is not particularly limited. For example, the reactant solution containing the compound represented by the general formula (3) can be introduced into the pressurized flow reaction channel. This reactant solution contains an acidic compound as a catalyst. In the reactant solution introduced into the pressurized flow reaction channel, the acidic compound is usually dissolved in the reactant solution. Alternatively, a solution containing the compound represented by the general formula (3) and a solution containing the acidic compound may be introduced into separate pressurized flow reaction channels and merged downstream to produce a reactant solution containing the compound represented by the general formula (3) and the acidic compound, and this reactant solution may be sent further downstream to the heating zone (H1) described below.

[0025] The content of the compound represented by the general formula (3) in the reaction raw material solution is preferably 10.0 to 300.0 mM, more preferably 20.0 to 280.0 mM, even more preferably 50.0 to 250.0 mM, and still more preferably 100.0 to 200.0 mM.

[0026] The content of the acidic compound in the reaction raw material solution is preferably 0.1 to 10.0M, more preferably 0.2 to 8.0M, even more preferably 0.4 to 6.0M, still more preferably 0.6 to 4.0M, and even more preferably 0.7 to 3.0M.

[0027] The solvent constituting the reaction raw material solution usually contains water. The proportion of water in the solvent constituting the reaction raw material solution is preferably 30% by mass or more, more preferably 50% by mass or more, even more preferably 70% by mass or more, still more preferably 80% by mass or more, and preferably 90% by mass or more, and it is also preferable that the solvent is entirely water. When the above solvent contains a solvent other than water, the solvent other than water is preferably a solvent miscible with water. Examples include alcohol solvents and nitrile solvents.

[0028] The size of the pressurized flow reaction channel of the present invention can be appropriately set depending on the reaction scale, etc. For example, the equivalent diameter can be 0.1 to 50.0 mm, 0.3 to 20.0 mm, 0.4 to 10.0 mm, or 0.5 to 5.0 mm. "Equivalent diameter" is also called equivalent (straight) diameter and is a term used in the field of mechanical engineering. When an equivalent circular pipe is assumed for a pipe or flow channel with an arbitrary internal cross-sectional shape, the diameter of the internal cross-section of that equivalent circular pipe is called the equivalent diameter. The equivalent diameter (deq) is defined as deq = 4A / p, where A is the internal cross-sectional area of ​​the pipe and p is the wetted edge length (inner circumference) of the pipe. When applied to a circular pipe, this equivalent diameter corresponds to the diameter of the internal cross-section of the circular pipe. The equivalent diameter is used to estimate the flow or heat transfer characteristics of the pipe based on data on the equivalent circular pipe and represents the spatial scale (representative length) of the phenomenon. The equivalent diameter is deq = 4a for a square pipe with a side a of the pipe cross section. ２ / 4a = a, and for an equilateral triangle tube with one side a, deq = a / 3 １／２ (See, for example, "Mechanical Engineering Dictionary," edited by the Japan Society of Mechanical Engineers, 1997, Maruzen Co., Ltd.)

[0029] The length of the pressurized flow reaction channel of the present invention can be appropriately set depending on the purpose and taking into consideration the desired reaction time, cooling time, etc. The material of the pressurized flow reaction channel of the present invention is not particularly limited, but a material that is resistant to corrosion under acidic conditions is preferred. For example, a resin tube can be used. Examples of resin tubes include tubes made of perfluoroalkoxyalkane (PFA), Teflon (registered trademark), aromatic polyether ketone resins, etc. Furthermore, stainless steel tubes or columns can also be used, as described below.

[0030] The compound represented by the general formula (3) is hydrolyzed to form R ５ is eliminated, and if necessary, neutralization treatment (quenching treatment) is carried out to produce a ribose phosphate compound represented by the following general formula (1).

[0031]

[0032] R １ , R ２ , R ３ and R １１ are R in the above general formula (3), respectively. １ , R ２ , R ３ and R １１ Immediately after the hydrolysis reaction carried out in the presence of an acidic compound, the group represented by general formula (2) of the ribose phosphate compound represented by general formula (1) is R １２ is a hydrogen atom. After the hydrolysis reaction, for example, by subjecting the compound to a neutralization treatment using an alkali metal hydroxide solution, R １２ A part or all of the above becomes alkali metal ions.

[0033] In the pressurized flow type reaction of the present invention, a preferred embodiment is to heat a solution containing the compound represented by general formula (3) and an acidic compound flowing through a pressurized flow type reaction channel to cause the hydrolysis reaction, then cool the reaction solution in which the hydrolysis reaction has occurred, and then warm the cooled reaction solution.

[0034] The heating step can promote hydrolysis, allowing the hydrolysis reaction to proceed sufficiently in a short period of time. From the viewpoints of improving the efficiency of the hydrolysis reaction and suppressing decomposition of the ribose phosphate compound produced, the heating temperature is preferably 105 to 160°C, more preferably 110 to 155°C, even more preferably 115 to 150°C, even more preferably 120 to 150°C, even more preferably 125 to 145°C, and even more preferably 130 to 140°C. The heating time (the hydrolysis reaction time, corresponding to the time spent flowing through the heating zone (H1) described below) is preferably 5 to 500 seconds, more preferably 10 to 450 seconds, even more preferably 15 to 400 seconds, even more preferably 20 to 350 seconds, even more preferably 25 to 300 seconds, even more preferably 30 to 250 seconds, even more preferably 35 to 200 seconds, even more preferably 40 to 180 seconds, even more preferably 50 to 120 seconds, and even more preferably 55 to 80 seconds.

[0035] The cooling step after the heating is a treatment to suppress thermal decomposition of the ribose phosphate compound produced by the hydrolysis reaction. This cooling is preferably performed at a temperature that does not freeze the solution. For example, cooling to a range of 0 to 40°C is preferred, more preferably to a range of 0 to 35°C, even more preferably to a range of 1 to 25°C, even more preferably to a range of 2 to 20°C, even more preferably to a range of 3 to 15°C, and even more preferably to a range of 6 to 15°C. Because the present invention employs a flow reaction, the temperature of the liquid flowing through the channel can be quickly controlled to the desired temperature. Therefore, the cooling step makes it possible to rapidly cool the reaction solution heated to a high temperature range in the heating step. Depending on the size of the channel, the reaction solution can be cooled to the desired temperature in a cooling time of, for example, 1 to 30 seconds. The cooling time is preferably 2 to 25 seconds, more preferably 3 to 20 seconds, and even more preferably 3 to 15 seconds, from the viewpoint of more reliably and quickly cooling to the desired temperature.

[0036] The heating step after the cooling is a treatment to prevent the precipitation of solutes in the reaction solution after passing through the outlet of the pressurized flow type reaction channel. ５ The bases (such as adenine, guanine, cytosine, uracil, or thymine) may become supersaturated in the reaction solution. For example, if the concentration of the compound represented by the general formula (3) in the raw solution is increased to, for example, 50 mM or more in order to increase the production efficiency of ribose phosphate compounds, the supersaturated state occurs after the hydrolysis reaction or the subsequent cooling step. A supersaturated reaction solution is unlikely to precipitate in a pressurized flow-type reaction channel; however, the release of pressure when passing through the outlet of the channel acts as an external stimulus, causing solute precipitation in the downstream piping. After extensive research to suppress this precipitation, the inventors discovered that by heating a supersaturated reaction solution subjected to a cooling step after heating just before the outlet of the pressurized flow-type reaction channel, the precipitation of bases when the pressure is released downstream of the outlet can be suppressed. From the viewpoint of obtaining a supersaturated reaction solution, the concentration of the compound represented by the general formula (3) in the raw solution is preferably 50 to 300 mM, more preferably 100 to 200 mM. By the heating, the temperature of the cooled reaction solution is preferably raised to a range of 10 to 60°C, more preferably to a range of 15 to 50°C, even more preferably to a range of 18 to 40°C, and even more preferably to a range of 20 to 30°C. The temperature in the cooling step is set lower than the temperature in the heating step. The difference between the temperature in the cooling step and the temperature in the heating step (the value obtained by subtracting the cooling temperature from the heating temperature) is preferably 5 to 50°C, more preferably 7 to 40°C, even more preferably 10 to 30°C, even more preferably 12 to 25°C, and even more preferably 14 to 20°C. The heating time is not particularly limited as long as it can lead to a temperature rise to the desired temperature range. The heating time can be, for example, 1 to 30 seconds, preferably 2 to 25 seconds, more preferably 2 to 20 seconds, and even more preferably 3 to 15 seconds.

[0037] Such heating, cooling and warming can be carried out, for example, by arranging a thermostatic bath for heating, a thermostatic bath for cooling and a thermostatic bath for warming from the upstream side to the downstream side of the pressure flow type reaction channel.

[0038] A preferred embodiment of the pressurized flow reaction system for carrying out the pressurized flow reaction of the present invention will be described with reference to FIG. 1. The pressurized flow reaction system shown in FIG. 1 has a pressurized flow reaction flow path having a raw material supply flow path (L1), a heating column (C1) arranged in the heating zone (H1), a cooling flow path (L2) arranged in the cooling zone (C), a heating flow path (L3) arranged in the heating zone (H2), and a back-pressure valve (B1) arranged downstream thereof. A pipe (L4) for withdrawing the reaction solution is connected downstream of the back-pressure valve (B1). The heating column (C1) can be, for example, a stainless steel column (flow path) that has excellent thermal conductivity and heat resistance. The inner surface of the stainless steel heating column is preferably subjected to a deactivation treatment such as Inert Mask to prevent corrosion by acid. A solution containing the compound represented by general formula (3) and an acidic compound introduced from the end of the raw material supply flow path (L1) is heated while flowing through the heating section column (C1) arranged in the heating zone (H1), causing a hydrolysis reaction. The reaction solution resulting from the hydrolysis reaction is cooled to a desired temperature while flowing through the cooling flow path (L2) arranged in the cooling section (C), thereby suppressing thermal decomposition of the ribose phosphate compound produced by the hydrolysis reaction. The cooled reaction solution is heated to a desired temperature while flowing through the heating flow path (L3) arranged in the heating zone (H2). The heated reaction solution is less likely to precipitate when it passes through the backpressure valve (B1) and the pressure is released, even if the base produced by the hydrolysis reaction is in a supersaturated state. As a result, clogging of the piping (L4) is less likely to occur, making it possible to continuously obtain the target ribose phosphate compound for a long period of time. The reaction solution taken out from the pipe (L4) can be adjusted to a neutral pH by adding an alkali if necessary, and the ribose phosphate compound can be purified by a conventional method such as column chromatography.

[0039] In a pressurized flow reaction, the flow rate of the liquid flowing through the pressurized flow reaction channel can be adjusted appropriately, taking into consideration the reaction time, etc., and also the channel length, etc. For example, the flow rate can be set to 0.1 to 100 mL / min, or alternatively, 0.2 to 50 mL / min, 0.5 to 20 mL / min, or 1.0 to 10 mL / min.

[0040] [Method for preventing solute precipitation in a supersaturated reaction solution] In relation to the technology relating to the manufacturing method of the present invention described above, the present invention provides the following method for preventing solute precipitation in a supersaturated reaction solution (hereinafter also referred to as the "precipitation prevention method of the present invention").

[0041] A method for preventing solute precipitation in a supersaturated reaction solution, comprising: when a supersaturated reaction solution flowing through a pressurized flow type reaction channel passes through an outlet of the pressurized flow type reaction channel and is released from pressure, adjusting the temperature of the supersaturated reaction solution upstream of the outlet to prevent solute precipitation downstream of the outlet.

[0042] The pressure of the pressurized flow-type reaction channel can be controlled, for example, by a back pressure valve. The temperature adjustment is preferably performed by increasing the temperature of the supersaturated reaction solution to a range of 10 to 60°C. That is, the heating step described above is preferred. Therefore, the temperature adjustment is preferably performed by increasing the temperature of the supersaturated reaction solution to a range of 15 to 50°C, more preferably to a range of 18 to 40°C, and even more preferably to a range of 20 to 30°C. The temperature of the supersaturated reaction solution before the temperature adjustment is preferably in the range of 0 to 40°C, more preferably in the range of 0 to 35°C, even more preferably in the range of 1 to 25°C, even more preferably in the range of 2 to 20°C, even more preferably in the range of 3 to 15°C, and even more preferably in the range of 6 to 15°C. Other preferred embodiments of the pressurized flow-type reaction channel can be those described in the production method of the present invention. Furthermore, the solvent in the supersaturated reaction solution can also be the solvent in the raw material solution described in the production method of the present invention. Examples of chemical reactions that produce the above-mentioned supersaturated reaction solution include hydrolysis, dephosphorylation, dehydration, neutralization, and oxidation of the compound represented by the above-mentioned general formula (3).

[0043] The present invention will be described in more detail based on examples, but the present invention should not be construed as being limited to these examples except as defined in the present invention.

[0044] Example 1 A ribose phosphate compound was obtained by carrying out the production method of the present invention using a pressurized flow reaction system shown in Figure 1. Details are shown below.

[0045] Liquid delivery pump (not shown): UI-22-110P manufactured by From Co., Ltd. was used, and a pressure sensor (PTCX-M6-PK2-50-SC) manufactured by Surpass Industrial Co., Ltd., a flow meter (NTFZ-1 / 4UNF-5-PK) manufactured by Surpass Industrial Co., Ltd., and a back pressure regulator (6010-57003) manufactured by IDEX were sequentially installed on the flow outlet side.

[0046] Heating zone (H1): An oil bath (O-1) manufactured by Yazawa Scientific Co., Ltd. was used and set to 135°C.

[0047] Raw material supply path (L1): A polyether ether ketone (PEEK) tube having an outer diameter of 1 / 16 inch, an inner diameter of 0.5 mm, and a length of 50 cm was used.

[0048] Heated column (C1): An InertMask-treated stainless steel column (inner diameter 4.6 mm x length 150 mm) was used.

[0049] Cooling zone (C): A small tabletop low-temperature water bath CBi-270A manufactured by AS ONE was used, and the temperature was set to 10°C.

[0050] Cooling flow path (L2): An InertMask-treated SUS316 tube having an outer diameter of 1 / 16 inch, an inner diameter of 0.8 mm and a length of 100 cm was used.

[0051] Heating zone (H2): A small tabletop low-temperature water bath CBi-270A manufactured by AS ONE was used and set at 25°C.

[0052] Heating flow path (L3): A PEEK tube with an outer diameter of 1 / 16 inch, an inner diameter of 0.5 mm, and a length of 100 cm was used.

[0053] Back pressure valve (B1): A back pressure regulator (6010-57005) manufactured by IDEX was used.

[0054] Piping (L4): A polytetrafluoroethylene (PTFE) tube having an outer diameter of 1 / 8 inch, an inner diameter of 1.58 mm, and a length of 100 cm was used.

[0055] Reaction raw material solution: 193 mL of 1 mol / L sulfuric acid (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and 10.5 g of 5'-adenylic acid (AMP, manufactured by Tokyo Chemical Industry Co., Ltd.) as a raw material were added to a 200 mL recovery flask and dissolved to prepare a reaction raw material solution.

[0056] Liquid delivery conditions: 2.00 mL / min, pressure in the pressurized flow reaction channel: 2.5 MPa

[0057] Flow time through the flow path: Heating column (C1): 75 seconds Cooling flow path (L2): 8 seconds Heating flow path (L3): 4 seconds

[0058] Withdrawal: 170 mL of a solution (reaction solution) containing ribose 5-phosphate was collected from the outlet of the pipe (L4) (95 minutes), and 24 mL of a 50% aqueous sodium hydroxide solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was added to adjust the pH to 7.0. The resulting aqueous solution was purified by column chromatography (packing material: Sephadex LH-20 (manufactured by Pharmacia), developing solvent: water / methanol), and then freeze-dried to obtain 6.29 g (85% yield) of ribose 5-phosphate disodium salt as the product. The yield was calculated using the following formula: Yield (%) = (mass of product) / (molar concentration of reaction raw material solution × molecular weight of product × volume of collected reaction liquid) × 100

[0059] Examples 2 to 6 Ribose 5-phosphate disodium salt was obtained in the same manner as in Example 1, except that the reaction conditions were changed as shown in the table below.

[0060] [Comparative Example 1] 0.435 g of AMP and 50 mL of 1 mol / L sulfuric acid were added to a 100 mL recovery flask and stirred at 100°C for 1 hour. HPLC confirmed that the AMP peak had disappeared. After cooling the recovery flask to 5°C, 56 mL of a 1 mol / L aqueous sodium hydroxide solution was added to adjust the pH to 7.0. After vacuum concentration, １ H-NMR analysis revealed that the desired ribose-5-phosphate disodium salt was not obtained.

[0061] The results of Examples 1 to 6 and Comparative Example 1 are shown in the table below. In the table, "s" stands for seconds.

[0062]

[0063] As shown in the table above, when the reaction raw material solution was hydrolyzed by a batch-type reaction under atmospheric pressure, the reaction temperature could not be raised above 100°C. In a reaction at 100°C under atmospheric pressure, hydrolysis of the raw material for, for example, a few minutes did not proceed sufficiently, resulting in a significantly low yield. Furthermore, when the reaction was allowed to proceed for 60 minutes as shown in the table, the resulting ribose 5-phosphate decomposed, making it impossible to obtain a ribose 5-phosphate compound (Comparative Example 1). In contrast, when the hydrolysis reaction was carried out in a pressurized flow reaction system, the desired ribose 5-phosphate compound could be obtained in high yield. In Example 6, a continuous reaction caused clogging of the piping after 60 minutes, but ribose 5-phosphate disodium salt could be obtained at a yield of approximately 85% until the clogging occurred.

[0064] While the present invention has been described in connection with embodiments thereof, we do not intend to limit our invention to any of the details of the description unless otherwise specified, and believe that the claims should be construed broadly without departing from the spirit and scope of the invention as set forth in the appended claims.

[0065] This application claims priority based on Japanese Patent Application No. 2024-098744, filed on June 19, 2024, the contents of which are incorporated herein by reference as part of the present specification.

Claims

1. A method for producing a ribose phosphate compound, comprising hydrolyzing a compound represented by the following general formula (3) through a pressurized flow reaction in the presence of an acidic compound to produce a ribose phosphate compound represented by the following general formula (1): In general formula (1), R １ is a hydrogen atom, a hydroxy group, or OR １１ indicates R １１ represents a group represented by the following general formula (2): ２ and R ３ each independently represents a hydrogen atom or a group represented by the following general formula (2): In the general formula (2), two R １２ Each independently represents a hydrogen atom or an alkali metal ion. * represents a link to an oxygen atom. In general formula (3), R １ , R ２ , R ３ and R １１ are R in the general formula (1). １ , R ２ , R ３ and R １１ It is synonymous with R ５ represents an adenyl group, a guanyl group, a cytosyl group, a uracil group, or a thyminyl group, provided that the compounds represented by the general formulas (1) and (3) have at least one group represented by the general formula (2) in the molecule.

2. The method for producing a ribose phosphate compound according to claim 1, wherein the solvent for the pressurized flow reaction comprises water.

3. The method for producing a ribose phosphate compound according to claim 2, wherein the acidic compound has a pH of 2.0 or less when made into an aqueous solution at a concentration of 1 mol / L.

4. The method for producing a ribose phosphate compound according to claim 3, wherein the acidic compound has a sulfo group.

5. The method for producing a ribose phosphate compound according to claim 2, wherein the pressurized flow reaction involves heating a solution containing the compound represented by general formula (3) flowing through a flow channel to cause the hydrolysis reaction, then cooling the reaction solution in which the hydrolysis reaction has occurred, and then warming the cooled reaction solution.

6. The method for producing a ribose phosphate compound according to claim 5, wherein the temperature of the solution is increased to a range of 105 to 160°C by heating, the temperature of the reaction solution is decreased to a range of 0 to 40°C by cooling, and the temperature of the reaction solution is increased to a range of 10 to 60°C by heating.

7. The method for producing a ribose phosphate compound according to claim 6, wherein the solution is heated for 30 to 300 seconds, and cooled for 1 to 30 seconds.

8. The method for producing a ribose phosphate compound according to claim 7, wherein the pressure of the pressurized flow reaction is 0.3 to 20.0 MPa.

9. The method for producing a ribose phosphate compound according to claim 8, wherein the pressure in the pressurized flow reaction is controlled by a back pressure valve.

10. The above R １ is a hydrogen atom or a hydroxy group, ２ is a hydrogen atom, ３ The method for producing a ribose phosphate compound according to claim 8, wherein: is a group represented by the general formula (2).

11. A method for preventing solute precipitation in a supersaturated reaction solution, comprising: when a supersaturated reaction solution flowing through a pressurized flow-type reaction channel passes through an outlet of the pressurized flow-type reaction channel and is released from pressure, adjusting the temperature of the supersaturated reaction solution upstream of the outlet to prevent solute precipitation downstream of the outlet.

12. The method for preventing solute precipitation in a supersaturated reaction solution according to claim 11, wherein the pressure in the pressurized flow type reaction channel is controlled by a back pressure valve.

13. The method for preventing solute precipitation in a supersaturated reaction solution according to claim 12, wherein the temperature adjustment is performed by increasing the temperature of the supersaturated reaction solution to a range of 10 to 60°C.

14. The method for preventing solute precipitation in a supersaturated reaction solution according to claim 13, wherein the temperature of the supersaturated reaction solution before the temperature adjustment is in the range of 0 to 40°C.

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