Process for the bulk production of sodium taurodeoxycholate

Abstract

The present application relates to a bulk production method of sodium taurodeoxycholate, which uses isopropyl alcohol to commercialize the production of sodium taurodeoxycholate, and separates and purifies by using a batch washing method, so that the process is simple, by-products are few, and impurities are few.

Description

Technical Field

[0001] This invention relates to a method for the mass production of sodium taurodeoxycholate, and more specifically, to a method for the mass production of sodium taurodeoxycholate, comprising: step 1), synthesizing crude sodium taurodeoxycholate; step 2), washing and filtering the crude sodium taurodeoxycholate synthesized in step 1) with an organic solvent to obtain a filter cake; and step 3), mixing the filter cake obtained in step 2) with a solution containing isopropyl alcohol, i) dissolving it by heating and stirring, ii) recrystallizing it by cooling and stirring, and iii) washing and filtering with isopropyl alcohol. Background Technology

[0002] Bile acids, or bile salts, are substances secreted by the liver and gallbladder, including glycocholic acid and taurocholic acid, which are analogous to substances with mechanisms of action similar to cholesterol. There are many types of bile acids, including cholic acid, chenodeoxycholic acid, taurocholic acid, and lithocholic acid. Studies have shown that these substances are helpful in the treatment and prevention of diseases.

[0003] Taurine dioxocholic acid is a bile acid, and almost the only naturally occurring sulfonic acid in its sulfonic form. In the case of sodium tauride deoxycholate, a salt form of taurine dioxocholic acid, it acts as a GPCR19 agonist, and can be used as an atopic prophylactic or therapeutic composition by reducing blood IgE levels, decreasing TH2 cytokine levels, and increasing TH1 cytokine levels (Korean Patent No. 10-1998402). It also has inhibitory or therapeutic effects on Alzheimer's disease and dementia by improving cognitive and behavioral disorders, inhibiting brain tissue apoptosis, enhancing immunity, and reducing the formation of β-amyloid emboli (Korean Patent No. 10-1743960).

[0004] However, for sodium tauride-deoxycholate to be practically developed into a pharmaceutical product, mass production is essential. In the mass production of sodium tauride-deoxycholate, difficulties have consistently arisen due to various unavoidable byproducts, unreacted raw materials, and the lack of significant differences in water solubility and dissolution during synthesis, hindering effective separation and purification. Furthermore, even when purification is carried out using well-known methods, issues such as recovery, reuse, yield, and inconsistent separation processes make commercially viable reproducibility difficult.

[0005] Relatedly, Korean Patent No. 10-0396113 discloses a method for purifying tauroursodeoxycholic acid using a two-layer extraction method with an organic solvent and an acidic or alkaline aqueous solution. Korean Patent No. 10-2068381 discloses a method for preparing bile acid derivatives using bile acids, but in this case, RuCl3, NaIO4, and an acid are used. However, methods for synthesizing sodium tauroursodeoxycholate, purifying tauroursodeoxycholic acid, and achieving commercially reproducible mass production methods remain unclear.

[0006] Therefore, the inventors have proposed a new method for the mass production of sodium taurine dioxocholate. When isopropanol is used for separation and purification by batch washing, the process is simple, the purity is high, and it can be mass-produced, thus completing the present invention. Summary of the Invention

[0007] Technical issues

[0008] To address the challenges of large-scale production of sodium tauride deoxycholate and the complexity of the process, this invention provides a method for the large-scale production of sodium tauride deoxycholate. This method uses isopropanol for separation and purification via batch washing, resulting in a simple process with fewer byproducts and higher purity.

[0009] Problem-solving methods

[0010] To address the aforementioned problems, this invention provides a method for the mass production of sodium taurodeoxycholate, comprising: step 1), synthesizing crude sodium taurodeoxycholate; step 2), washing and filtering the crude sodium taurodeoxycholate synthesized in step 1) using an organic solvent to obtain a filter cake; and step 3), mixing the filter cake obtained in step 2) with a solution containing isopropyl alcohol, i) dissolving it by heating and stirring, ii) recrystallizing it by cooling and stirring, and iii) washing and filtering it with isopropyl alcohol.

[0011] Furthermore, a sodium taurine deoxycholate prepared by the above-described mass production method is provided.

[0012] In this invention, the batch production method of sodium tauride deoxycholate can produce more than 1 kg of sodium tauride deoxycholate each time.

[0013] In one embodiment of the present invention, the crude sodium tauride deoxycholate in step 1) above is synthesized by placing a solution containing sodium tauride, deoxycholic acid and N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) into a stirrer and stirring while adjusting the temperature.

[0014] In one embodiment of the present invention, the organic solvent in step 2) is selected from one or more of the group consisting of ethanol, acetone, pyridine, hexafluoroisopropanol, propanol, butanol, cyclohexane, toluene, dichloromethane, diethyl ether, ethyl acetate, methyl acetate, and mixtures of two or more of these solvents.

[0015] In one specific embodiment of the present invention, the organic solvent in step 2) above includes ethanol and acetone, and the organic solvent is a mixed solvent of ethanol and acetone in a volume ratio of 1:0.5 to 2.

[0016] In one embodiment of the present invention, step 3) is repeated more than twice. In a specific embodiment of the present invention, step 3) is repeated two to three times.

[0017] In one embodiment of the present invention, the isopropyl alcohol-containing solution in step 3) above comprises water and isopropyl alcohol, wherein the volume ratio of water to isopropyl alcohol is 1:1 to 10.

[0018] In one embodiment of the present invention, the heating in step 3)i) above is to heat to 20°C to 100°C.

[0019] Furthermore, in one embodiment of the present invention, the cooling in step ii) of step 3) above is cooling to 0°C to 50°C.

[0020] Furthermore, in one embodiment of the present invention, the stirring in step ii) of step 3) above is carried out for 8 to 30 hours.

[0021] Furthermore, in one embodiment of the present invention, the isopropyl alcohol in the solution containing isopropyl alcohol in step 3) is 5 to 20 times the weight of the filter cake in step 2).

[0022] In one embodiment of the invention, step 4) is further included, in which sodium taurine deoxycholate obtained in step 3) is purified using a mixed solution containing acetone.

[0023] In one specific embodiment of the present invention, step 4) above may include: step A), dissolving the sodium tauride deoxycholate filtered in step 3) in a mixed solution containing acetone; step B), adding acetone dropwise to the solution in step A), cooling and stirring to recrystallize; and step C), washing the recrystallized sodium tauride deoxycholate with acetone, filtering and drying.

[0024] The effects of the invention

[0025] This invention relates to a method for the mass production of sodium tauride deoxycholate, which uses isopropanol for purification through batch washing. Therefore, the process is simple, can be mass-produced, and has high purity, thus having the advantage of enabling the industrial production of sodium tauride deoxycholate. Attached Figure Description

[0026] Figure 1 This is a simplified diagram of the mass production method of the present invention.

[0027] Figure 2 A diagram illustrating the purification process in the mass production method of the present invention.

[0028] Figure 3 A diagram illustrating the scheme of the mass production method of the present invention.

[0029] Figure 4 The figure shows a chromatographic representation to confirm whether the sodium taurine deoxycholate prepared according to the production method of the present invention is synthesized.

[0030] Figure 5 The figure shows the purity of sodium taurine deoxycholate prepared according to the production method of the present invention, confirmed by HPLC. Detailed Implementation

[0031] Best way to carry out the invention

[0032] In the preparation of sodium taurodeoxycholate, it is produced in batches using a solution containing isopropyl alcohol.

[0033] Methods of implementing the present invention

[0034] The present invention will now be described in detail.

[0035] Throughout the entire specification of this invention, unless otherwise stated, when a portion “includes” a structural element, it means that other structural elements may also be included, rather than excluding other structural elements.

[0036] Unless otherwise defined in this specification, a compound generally refers to a recognized compound.

[0037] This invention relates to a method for the mass production of sodium tauride-deoxycholate, comprising: step 1), synthesizing crude sodium tauride-deoxycholate; step 2), washing and filtering the crude sodium tauride-deoxycholate synthesized in step 1) with an organic solvent to obtain a filter cake; and step 3), mixing the filter cake obtained in step 2) with a solution containing isopropyl alcohol, i) dissolving it by heating and stirring, ii) recrystallizing it by cooling and stirring, and iii) washing and filtering with isopropyl alcohol.

[0038] Furthermore, this relates to sodium taurine deoxycholate prepared by the aforementioned mass production method.

[0039] In this invention, the reactants used to prepare crude tauride deoxycholate sodium can be, for example, unprocessed deoxycholic acid, taurine, sodium taurate, tauride dioxycholic acid derivatives, etc. Furthermore, the reactants can be commercially available products, synthesized using methods known in the art, or obtained by processing samples collected from nature. This is illustrative and not limited to the aforementioned substances or methods.

[0040] Specifically, in one embodiment of the present invention, the crude sodium tauride deoxycholate obtained in step 1) above can be synthesized by placing a solution containing sodium tauride, deoxycholic acid and N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) into a stirrer and stirring while adjusting the temperature.

[0041] More specifically, synthetic sodium taurate and deoxycholic acid can be used. Sodium taurate can be prepared by mixing taurine and its sodium salt, and deoxycholic acid can be obtained by crystallizing natural deoxycholic acid, but is not limited thereto, including all sodium taurate and deoxycholic acid prepared by known methods or obtained commercially. Furthermore, in this invention, appropriate amounts of the above reactants can be used to mass-produce crude sodium taurodeoxycholate.

[0042] Furthermore, in this invention, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) is a compound represented by the following chemical formula, which can be represented by the IUPAC names "2-ethoxy-2H-quinoline-1-carboxylic acid ethyl ester" and "1,2-dihydro-2-ethoxyquinoline-1-carboxylate".

[0043]

[0044] Specifically, when sodium taurine, deoxycholic acid, and EEDQ are used in this invention, the following chemical reaction can be carried out.

[0045]

[0046] In this invention, step 1) above can be used to synthesize sodium tauride-deoxycholate by synthesizing reactants. According to a specific synthesis method of one embodiment of the present invention, deoxycholic acid and EEDQ (N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline) react to form an intermediate with quinoline as a byproduct. This intermediate then reacts with sodium taurate to form sodium tauride-deoxycholate. Furthermore, ethanol can be used as the solvent in this process.

[0047] More specifically, in this invention, when using sodium taurine, deoxycholic acid, and EEDQ for batch synthesis, in the first step (1) described above... st In step (2), it exhibits high reactivity, low side reactions, and high stability during the reaction at 40℃. Furthermore, in the aforementioned second step (2)... nd In step (1), not only does it possess the advantages of high reactivity, low side reactions, and stability during the reaction process, but the resulting solid sodium tauride deoxycholate can also be easily removed by solvent removal of free active groups. Furthermore, through the above purification process, sodium tauride deoxycholate with a single impurity of less than 0.1% can be obtained in high yield and with high purity.

[0048] In step 1) above, the amount of reactants, temperature, and stirring can be controlled at appropriate levels based on the amount of sodium taurine deoxycholate synthesized. Furthermore, to determine these appropriate levels, analysis of substances produced in step 1), unreacted substances, etc., can be used.

[0049] Specifically, step 1) above may include heating and stirring as well as cooling and stirring to synthesize sodium taurine deoxycholate, but is not limited thereto.

[0050] In the above synthesis steps, the heating temperature can range from 30°C to 100°C, and more specifically, it can be heated to 30°C to 80°C, 31°C to 70°C, 32°C to 60°C, 33°C to 55°C, 34°C to 50°C, or 35°C to 45°C while stirring. However, when the heating temperature exceeds 100°C, the amount of impurities generated can increase significantly, making purification impractical in terms of time and cost. Furthermore, when the heating temperature is around 40°C, side reactions are low and stability is high.

[0051] Furthermore, in the above synthesis steps, the heating and stirring step can be stirring for 10 to 30 hours, more specifically, for 12 to 28 hours or 14 to 26 hours. Also, specifically, crystals can be produced 0.5 to 5 hours after stirring begins, and the process may also include a step of releasing the produced crystals to prevent agglomeration and thus achieve a high yield in the synthesis.

[0052] Furthermore, in the above synthesis steps, the cooling temperature can be from 0°C to 30°C, more specifically, from 10°C to 30°C or from 15°C to 25°C. When the cooling temperature drops below 0°C, impurities increase; therefore, the temperature should be kept above 0°C. A cooling temperature of around 20°C is conducive to crystallization.

[0053] Furthermore, in the above synthesis steps, the cooling and stirring steps can be stirred for 0.1 hours to 5 hours, more specifically, the stirring time can be 0.2 hours to 3 hours, 0.3 hours to 2 hours, 0.4 hours to 2 hours, or 0.5 hours to 2 hours.

[0054] In this invention, step 2) involves washing and filtering the substance produced in step 1).

[0055] In one embodiment of the present invention, the organic solvent in step 2) is selected from one or more solvents chosen from the group consisting of ethanol, acetone, pyridine, hexafluoroisopropanol, propanol, butanol, cyclohexane, toluene, dichloromethane, diethyl ether, ethyl acetate, methyl acetate, and mixtures of two or more thereof. One or more of the above organic solvents may be selected as a single solvent, or a mixture of two or more solvents may be included.

[0056] In one specific embodiment of the present invention, the organic solvent in step 2) above comprises ethanol and acetone, wherein the organic solvent is a mixed solvent in which the volume ratio of ethanol to acetone is 1:0.5 to 2. More specifically, the organic solvent may be a mixed solvent in which the volume ratio of ethanol to acetone is 1:0.6 to 1.4, 0.75 to 1.25, or 0.8 to 1.2.

[0057] In this invention, step 3) involves purifying the substances obtained in steps 1) and 2). In this invention, step 3) can be purified by batch washing, so the substances obtained in steps 1) and 2) are purified without drying. Since this is not the existing column method, the process is simple, has fewer impurities, and can be carried out in batches, thus enabling the mass production of sodium taurine deoxycholate.

[0058] In one embodiment of the present invention, step 3) above may be repeated more than twice. In a specific embodiment of the present invention, step 3) above may be repeated two to three times. The number of repetitions of step 3) above can be selected considering purity and yield. Since there is no drying process between each repetition, purification can be achieved through a simple process.

[0059] The amount of isopropanol, temperature, and stirring time used in step 3) above can be adjusted appropriately according to the amount of sodium taurine deoxycholate synthesized.

[0060] In one specific embodiment of the present invention, the isopropyl alcohol-containing solution in step 3) above comprises water and isopropyl alcohol, and the mixing volume ratio of water to isopropyl alcohol can be from 1:1 to 10. In a more specific embodiment of the present invention, the mixing volume ratio can be from 1:5 to 10 or from 1:6 to 9.

[0061] In one specific embodiment of the invention, the heating in step 3)i) above is to 20°C to 100°C. Specifically, it can be heated to 30°C to 90°C, 35°C to 85°C, 40°C to 80°C, 45°C to 75°C, 46°C to 74°C, 47°C to 73°C, 48°C to 72°C, 49°C to 71°C, or 50°C to 70°C while stirring. The heating in step 3)i) can be achieved by completely dissolving the substance in a mixed solution containing isopropanol while stirring, and then stirring for another 10 to 30 minutes after confirming dissolution. Reactions at around 60°C show excellent purification time and stability, but are not limited to this.

[0062] In one specific embodiment of the present invention, the cooling in step ii) of step 3) above is cooling to 0°C to 50°C. Specific cooling temperatures can be 10°C to 40°C, 15°C to 35°C, 15°C to 34°C, 15°C to 33°C, 15°C to 32°C, 15°C to 31°C, or 15°C to 30°C. Cooling can be performed to form crystals. Rapid cooling prevents crystallization from proceeding smoothly, therefore slow cooling is preferable.

[0063] In one specific embodiment of the invention, the stirring in step ii) of step 3) above is carried out for 8 to 30 hours. More specifically, the stirring time can be 8 to 25 hours, 8 to 20 hours, or 9 to 15 hours. The stirring time is adjusted appropriately taking into account the amount of reactants and the temperature.

[0064] In one embodiment of the invention, the isopropyl alcohol in the solution containing isopropyl alcohol in step 3) above can be 5 to 20 times the weight of the filter cake in step 2) above. Specifically, the isopropyl alcohol used includes the total amount of isopropyl alcohol used in step 3), which, when mixed with water, refers to the weight of isopropyl alcohol excluding water. More specifically, the isopropyl alcohol weight ratio can be 5 to 15 times, 6 to 14 times, or 7 to 12 times the weight of the filter cake, within which the degree of recrystallization is most excellent.

[0065] In one embodiment of the present invention, step 4) may be included after step 3), which may be a step of purifying sodium taurine deoxycholate obtained in step 3) using a mixed solution containing acetone.

[0066] In one specific embodiment of the present invention, step 4) above may include: step A), dissolving the sodium tauride deoxycholate filtered in step 3) in a mixed solution containing acetone; step B), adding acetone dropwise to the solution in step A), cooling and stirring to recrystallize; and step C), washing the recrystallized sodium tauride deoxycholate with acetone, filtering and drying.

[0067] The present invention may also include step 4), therefore this step is not a necessary structural element of the present invention. When step 4) is further included, a purity of more than 99.5% can be achieved.

[0068] Furthermore, in one specific embodiment of the present invention, the acetone-containing mixed solution in step A) of step 4) above comprises water and acetone, and the mixing volume ratio of water to acetone is 1:1 to 10. In a more specific embodiment of the present invention, specifically, in the mixed solvent of step A) above, the mixing volume ratio of water to acetone can be 1:5 to 10, or 1:6 to 9.

[0069] Furthermore, in a specific embodiment of the present invention, in step 4) above, step B) of adding acetone dropwise to the solution in step A) and then cooling and stirring to recrystallize includes not only the steps of adding acetone dropwise to the solution and then stirring, adding acetone dropwise after stirring, or adding acetone dropwise while stirring, but also the steps of stopping the addition of acetone and stirring when crystals are generated by adding acetone dropwise and stirring, thus not being limited to the order of adding, cooling and stirring.

[0070] Furthermore, in a specific embodiment of the present invention, step C) of step 4) above can be performed at 10°C to 40°C, more specifically, at 15°C to 35°C, 15°C to 34°C, 15°C to 33°C, 15°C to 32°C, 15°C to 31°C, or 15°C to 30°C, with the purpose and effect of cooling as described above.

[0071] Furthermore, in one embodiment of the present invention, the acetone in step 4) can be 5 to 20 times the weight of the filter cake filtered in step 3), more specifically, it can be 5 to 15 times, 5.5 to 12 times, or 6 to 12 times. It also includes the amount of all acetone used in steps A) to C) of step 4), and when used in combination with water, it refers to the weight of acetone excluding water.

[0072] In one embodiment of the present invention, the batch production method of sodium tauride deoxycholate can produce more than 1 kg of sodium tauride deoxycholate each time.

[0073] Furthermore, in one embodiment of the present invention, the yield of sodium taurodeoxycholate can reach 25% or more. More specifically, the yield can be 25% or more, 26% or more, 27% or more, 28% or more, 29% or more, or 30% or more.

[0074] Furthermore, in one embodiment of the present invention, sodium tauride deoxycholate with a purity of 99% can be prepared by a mass production method for sodium tauride deoxycholate. More specifically, the purity can be 99.1% or higher, 99.15% or higher, 99.2% or higher, 99.25% or higher, 99.3% or higher, 99.35% or higher, 99.4% or higher, 99.45% or higher, or 99.5% or higher.

[0075] In this invention, the sodium tauride produced by the batch production method of sodium tauride has a yield of more than 25%, a purity of more than 99%, and can produce more than 1 kg each time, thereby effectively preparing sodium tauride.

[0076] The present invention will now be described in detail through examples and experimental cases.

[0077] However, the following embodiments and experimental examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following embodiments and experimental examples.

[0078] Example 1. Batch synthesis of sodium taurate

[0079] 2.04 kg of sodium hydroxide and 44 L of ethanol were added to the reactor and stirring was started. During stirring, the external temperature was set to 55°C to raise the internal temperature to 55°C. After confirming complete dissolution of the sodium hydroxide, 5.5 kg of taurine was added. Stirring was continued for at least 2 hours after the addition of taurine. After stirring, the external temperature was set to 30°C to cool the internal temperature to 30°C, and stirring was continued for 1 hour. After stirring, the mixture was washed / filtered using a Nutsche filter. Washing / filtration was first performed with 27.5 L of ethanol, followed by further washing / filtration with 16.5 L of acetone. The filtered filter cake (wet cake) was placed in a tray and placed in a dryer, then vacuum dried at 35°C for at least 12 hours to obtain 5.28 kg of sodium taurine.

[0080] Example 2. Batch synthesis of sodium taurodeoxycholate

[0081] 1.96 kg of sodium taurate (prepared in Example 1), 5.5 kg of deoxycholic acid, 6.93 kg of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), and 55 L of ethanol were added to the reactor, and stirring was initiated. During stirring, the external temperature was set to 40°C to raise the internal temperature to 40°C, and stirring was continued for at least 15 hours. After stirring, the external temperature was set to 20°C to cool the internal temperature to 20°C, and stirring was continued for 1 hour after cooling. After stirring, the mixture was washed / filtered using a Nutsche filter. The washing / filtration was performed twice using a mixture of 55 L of ethanol and acetone (mixed volume ratio 1:1), followed by washing / filtration with 16.5 L of acetone to obtain 7.515 kg of the first (1 st Filter cake.

[0082] Example 3-1. Batch purification of sodium taurodeoxycholate (1)

[0083] 7.515 kg of the first filter cake prepared in Example 2, 76.1 L of isopropyl alcohol, and 10.1 L of purified water were placed in the reactor and stirred. While stirring, the external temperature was set to 60°C and heated until complete dissolution was confirmed, then stirred for another 20 minutes. Afterward, the mixture was slowly cooled to 20°C over 3 hours, and then stirred for another 12 hours or more. After stirring, the mixture was washed / filtered using a Nutsche filter, and then washed / filtered with 15.6 L of isopropyl alcohol to obtain 13.13 kg of the second (2) filter cake. nd Filter cake.

[0084] Next, 13.13 kg of the second filter cake, 56.8 L of isopropanol, and 7.6 L of purified water were added to the reactor and stirred. While stirring, the external temperature was set to 60°C and heated until complete dissolution was confirmed, then stirred for another 20 minutes. The mixture was then slowly cooled to 20°C over 3 hours, and stirred for at least 12 hours. After stirring, the mixture was washed / filtered using a Nutsche filter, followed by washing / filtering with 11.4 L of isopropanol to obtain 7.88 kg of the third filter cake. rd Filter cake.

[0085] Next, 7.88 kg of the third filter cake, 42.6 L of isopropanol, and 5.7 L of purified water were added back to the reactor and stirred. While stirring, the external temperature was set to 60°C and heated until complete dissolution was confirmed, then stirred for another 20 minutes. The mixture was then slowly cooled to 20°C over 3 hours, followed by stirring for at least 12 hours. After stirring, the mixture was washed / filtered using a Nutsche filter, this time with 8.5 L of isopropanol, yielding a wet solid. The wet solid was weighed and measured to be 4.73 kg. The wet solid was then placed in a tray and placed in a desiccator, where it was vacuum dried at 35°C for at least 6 hours to obtain 2.84 kg of sodium tauride deoxycholate (purity 99.0%).

[0086] Example 3-2. Batch purification of sodium taurodeoxycholate (1)

[0087] Except for the difference in the weight of the first filter cake, sodium tauride was purified in batches using the same process as in Example 3-1 above. 6.91 kg of the first filter cake was added, followed by 11.34 kg of the second filter cake, 6.81 kg of the third filter cake, and 4.08 kg of the filtered solid (wet solid) to obtain 2.45 kg of sodium tauride (purity 99.9%).

[0088] Example 4. Batch purification of sodium taurodeoxycholate (2)

[0089] 2.84 kg and 2.45 kg of sodium taurodeoxycholate purified in Examples 3-1 and 3-2, 64.2 L of isopropyl alcohol, and 8.6 L of purified water were added to the reactor and stirred. While stirring, the external temperature was set to 60°C and heated until complete dissolution was confirmed, then stirred for another 20 minutes. Next, the mixture was slowly cooled to 20°C over 3 hours, and then stirred for at least 12 hours. After stirring, the mixture was washed / filtered using a Nutsche filter, this time with 12.8 L of isopropyl alcohol, yielding 6.89 kg of filter cake (wet cake).

[0090] Next, 6.89 kg of the filtered cake, 47.6 L of isopropanol, and 6.4 L of purified water were added to the reactor and stirred. While stirring, the external temperature was set to 60°C and heated until complete dissolution was confirmed, then stirred for another 20 minutes. The mixture was then slowly cooled to 20°C over 3 hours, followed by stirring for at least 12 hours. After stirring, the mixture was washed / filtered using a Nutsche filter with 9.7 L of isopropanol to obtain a wet solid. The weight of the wet solid was measured to be 5.17 kg. The wet solid was then placed in a tray and placed in a dryer, where it was vacuum dried at 35°C for at least 6 hours to obtain 3.10 kg of sodium tauride deoxycholate.

[0091] Example 5. Batch purification of sodium taurodeoxycholate (3)

[0092] 3.10 kg of sodium taurodeoxycholate obtained in Example 4, 2.9 L of acetone, and 2.9 L of purified water were added to the reactor and stirred. During stirring, the external temperature was set to 30°C to control the internal temperature at 30°C, ensuring complete dissolution. The resulting solution was then filtered through a 0.45 μm cartridge filter and transferred to a crystallizer.

[0093] Next, 8.3 L of acetone was added dropwise to the crystallizer. The addition was stopped when crystals formed, and the mixture was stirred for 1 hour. After stirring, 10 L of acetone was added dropwise while stirring. The mixture was then cooled to 20°C and stirred for another 2 hours. After stirring, the mixture was washed / filtered using a Nutsche filter with 7.2 L of acetone, yielding 5.92 kg of filtered filter cake (wet filter cake). The filtered filter cake was placed in a tray and then placed in a dryer. It was then vacuum dried at 70°C for at least 12 hours to obtain 2.61 kg of crystallized sodium taurine deoxycholate. (Yield: 25 ± 2%, purity ≥ 99.9%)

[0094] Example 6. Freeze-drying of sodium taurodeoxycholate

[0095] 2.61 kg of the crystallized sodium tauride obtained in Example 5 was dissolved in 25.0 L of distilled water to prepare a solution, which was then placed in a tray and put into a freeze dryer. Next, the freeze dryer was set up as shown in Table 1, and freeze drying was performed. 2.57 kg of crystals were obtained by freeze drying. The freeze-dried crystals were then pulverized for 90 minutes in a pulverizer with a mesh size of 0.99 mm and a rotation speed of 3000 rpm to obtain crystallized sodium tauride.

[0096] Table 1

[0097]

[0098] Experimental Example 1. Confirmation of the synthesis of sodium taurine deoxycholate.

[0099] The sodium tauride-deoxycholate synthesized in Example 2 above was sampled and analyzed by HPLC. HPLC analysis was performed under the conditions shown in Table 2. The mobile phase was prepared by mixing 4.0 g of sodium dihydrogen phosphate monohydrate, 0.606 g of sodium dodecyl sulfate, 1000 ml of water, and 515 ml of acetonitrile, and adjusting the pH to 2.1 with phosphoric acid. 50 mg was added to a 10 ml flask, dissolved in 3.0 ml of methanol, and then filled to the mark with the mobile phase. The sample was filtered through a 0.45 μm membrane syringe filter and placed in an HPLC vial. The analytical results confirmed the synthesis of sodium tauride-deoxycholate.

[0100] Table 2

[0101] object condition Detector 220nm Column Cl8, 5μm, 4.6mm(ID)*250mm(length) Temperature 40℃ Flow rate 1.0 ml / min Injection Volume 30μl Run time 105min

[0102] Experimental Example 2. Confirmation of Unreacted Substances and Detection of Sodium Taurodeoxycholate

[0103] In Example 2 above, the ethanol solution in the supernatant after crystal precipitation was sampled, and the presence of reactants was confirmed by TLC. The TLC developing solvents were: 1) CHCl3:MeOH = 9:1, PMA as the staining solvent for deoxycholic acid; 2) IPA:H2O = 8:2, ninhydrin as the staining solvent for taurine; and 3) CH3Cl:MeOH = 7.5:2.5 as the staining solvent for sodium taurodeoxycholate. Furthermore, EEDQ and quinoline as a byproduct were visually confirmed under UV 254 nm. The analytical results are as follows: Figure 4 As shown, it was confirmed that deoxycholic acid, which is the starting material, was not present.

[0104] Experimental Example 3. Confirmation of the purity of sodium taurine deoxycholate based on the number of purification (recrystallization) cycles. Experimental Example 3-1. Impurity detection test.

[0105] To confirm the purity of the sodium taurine deoxycholate prepared according to the above examples, it was necessary to confirm the detection of sodium deoxycholate, related substances, heavy metals, taurine, residual solvents, and drying loss.

[0106] During testing, a sample solution of sodium taurodeoxycholate dissolved in a solvent and a standard solution of sodium taurodeoxycholate dissolved in a solvent, prepared according to the examples, were prepared and measured using a UV spectrophotometer. Furthermore, the loss on drying was determined according to the loss on drying test method in the general test methods of the Korean Pharmacopoeia, and heavy metals were determined according to the first method of heavy metal test method in the general test methods of the Korean Pharmacopoeia.

[0107] The analysis results are shown in Tables 3 to 8 below.

[0108] Table 3

[0109]

[0110]

[0111] Table 4

[0112]

[0113] Table 5

[0114]

[0115]

[0116] Table 6

[0117]

[0118] Table 7

[0119]

[0120] Table 8

[0121]

[0122]

[0123] As shown in Tables 3 to 8, it was confirmed that the sodium taurine deoxycholate synthesized in batches by the purification method of the present invention has very high purity due to the very low content of sodium deoxycholate, related substances, heavy metals, etc.

[0124] Experiment Example 3-2. Purity Experiment

[0125] The purified sodium taurine deoxycholate was sampled at various stages according to Examples 2 to 5 above, and its purity was analyzed using the method described above. The analytical results are shown in Table 9.

[0126] Table 9

[0127]

[0128] As shown in Table 9, it was confirmed that sodium tauride-deoxycholate was produced in batches using the purification (recrystallization) method of the present invention, and the purity was very high. Specifically, Examples 3-1, 3-2, 4, and 5 above each correspond to one purification (recrystallization). When the recrystallization was performed two or more times, the purity was equivalent to more than 99%, and the yield was also as high as more than 25%. Therefore, it was confirmed that sodium tauride-deoxycholate can be produced in batches.

[0129] Comparative example: Synthesis of sodium taurine deoxycholate according to other synthetic methods.

[0130] 1-1. Synthesis using isobutyl chloroformate

[0131] The synthesis was carried out using the same process as in Example 2 above. Sodium taurate, deoxycholic acid (DCA), and isobutyl chloroformate were placed in a reactor, and sodium taurate was synthesized by changing the temperature, solvent, and base. The synthesis process was carried out through the following chemical reactions.

[0132]

[0133] The above synthesis method suffers from poor reaction in the first step. By changing the temperature and / or solvent, the reaction conditions for quantitative conversion of DCA are found. Then, in the second step, due to the low reaction rate, the reaction is carried out by changing the solvent, temperature, and base, but the yield is significantly lower.

[0134] 1-2. Synthesis using Pivaloyl chloride (trimethylacetyl chloride)

[0135] The synthesis was carried out using the same process as in Example 2 above, with sodium taurate, deoxycholic acid (DCA), and pivaloyl chloride placed in a reactor, and sodium taurate was synthesized by changing the temperature and / or solvent. The synthesis process was carried out through the following chemical reactions.

[0136]

[0137] It was confirmed that the DCA used as the starting material in the first step of the above synthesis method was not completely converted, and the conversion rate to TDC in the second step was less than 50%. Although the reaction solvent, reaction temperature, and base were changed, the reaction rate was still low.

[0138] l-3. Synthesis using TPP / DTBT

[0139] The synthesis was carried out using the same process as in Example 2 above. Sodium taurate, deoxycholic acid, triphenylphosphine (TPP), and DTBT were placed in a reactor, and sodium taurate was synthesized by varying the temperature, solvent, and alkali. The synthesis process involved the following chemical reactions.

[0140]

[0141] It was confirmed that the above synthetic method resulted in a rapid reaction for preparing the activating group in the first step, but the subsequent reaction with DCA was incomplete. Furthermore, residual starting materials DCA and TPP / DTFT salts with similar solubility were difficult to remove completely after the reaction, leading to a reaction rate of only 20-30% in the second step and a decreased yield.

[0142] 1-4. Synthesis using DCC / HOBt / MMP

[0143] The synthesis was carried out using the same process as in Example 2 above. Sodium taurate, deoxycholic acid, N,N'-dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole (HOBt), and MMP were placed in a reactor, and sodium taurate was synthesized by varying the temperature and solvent. The synthesis process involved the following chemical reactions.

[0144]

[0145] It was confirmed that the above synthetic method completely converts the active ester into a complete product in the first step, and the active ester used as the starting material also completely disappears in the second step reaction. Although the reactivity is high, it generates a large amount of impurities. Furthermore, the work-up process is complex, and impurity removal is not smooth, resulting in a yield and purity of 65-70% and 70-75%, respectively, which is poor. The specific purity is shown in Table 10 below.

[0146] Table 10

[0147] RT Imurity Area (%) Remove recrystallization solvent 3.2min 17.75％ Cannot be removed EA wash (11 times) 4.8min 2-3％ Can remove <![CDATA[IPA: H2O recrystallization (4 times)]]>

[0148] 1-5. Synthesis using DSC

[0149] The synthesis was carried out using the same process as in Example 2 above. Sodium taurate, deoxycholic acid, and N,N'-disuccinimudyl carbonate (DSC) were placed in a reactor, and sodium taurate was synthesized by varying the temperature and solvent. The synthesis process involved the following chemical reactions.

[0150]

[0151] It was confirmed that the above synthetic method converts the product into a complete active ester in the first step of the TLC stage, but generates a large number of impurities in the second step. Furthermore, while the reaction proceeds easily at low temperatures, it is difficult to remove new impurities, resulting in a low yield and purity of 55–60% and 0.2%, respectively. Specific purity values ​​are shown in Table 11 below.

[0152] Table 11

[0153]

[0154] 1-6. Synthesis using HATU

[0155] The synthesis was carried out using the same process as in Example 2 above. Sodium taurate, deoxycholic acid, and HATU (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-Oxide Hexafluorophosphate) were placed in a reactor, and sodium taurate was synthesized by varying the temperature, solvent, and alkali. The synthesis process was carried out through the following chemical reactions.

[0156]

[0157] It has been confirmed that the above synthetic method converts the active ester into a complete product in the first step, and the active ester is also completely converted in the second step, thus exhibiting excellent reactivity. However, new impurities are also formed along with TDC in the related substances, and the byproducts are relatively difficult to remove. Furthermore, it is necessary to dissolve and remove the impurities and byproducts with H2O, but TDC also dissolves with H2O, thus presenting a problem of difficulty in separating them.

[0158] Industrial availability

[0159] This invention demonstrates that sodium taurine deoxycholate can be mass-produced and can be effectively utilized in chemical, pharmaceutical, and other industrial fields.

Claims

1. A method for mass production of sodium taurine deoxycholate, characterized in that, include: Step 1) A solution containing sodium taurine, deoxycholic acid and N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline is placed in a mixer and stirred while adjusting the temperature to synthesize crude sodium taurine. Step 2) The crude sodium taurine synthesized in step 1) is washed and filtered using an organic solvent to obtain a filter cake. Step 3), after mixing the filter cake obtained in step 2) with a solution containing isopropanol, i) dissolve it by heating and stirring, ii) recrystallize it by cooling and stirring, iii) wash it with isopropanol and filter it; as well as Step 4) Dissolve the sodium tauride deoxycholate filtered in step 3) in a mixed solution containing acetone; i) Add acetone dropwise to the solution, cool and stir to recrystallize; ii) Wash the recrystallized sodium tauride deoxycholate with acetone, filter and dry to purify.

2. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, The organic solvent in step 2) above is selected from one or more of the following groups: ethanol, acetone, pyridine, hexafluoroisopropanol, propanol, butanol, cyclohexane, toluene, dichloromethane, diethyl ether, ethyl acetate, methyl acetate, and mixtures of two or more of these solvents.

3. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, The organic solvent in step 2) above includes ethanol and acetone, and the organic solvent is a mixed solvent in which the volume ratio of ethanol to acetone is 1:0.5 to 2.

4. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, Repeat step 3) above at least twice.

5. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, Repeat step 3) above two to three times.

6. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, The isopropanol-containing solution in step 3) above contains water and isopropanol, with a mixing volume ratio of water to isopropanol of 1:1 to 10.

7. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, The heating in step 3)i) above is to heat to 20°C to 100°C.

8. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, The cooling in step ii) of step 3) above is to cool to 0°C to 50°C.

9. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, The stirring in step 3) ii) above shall be carried out for 8 to 30 hours.

10. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, The isopropanol in the solution containing isopropanol in step 3) above is 5 to 20 times the weight of the filter cake in step 2) above.

11. The method for mass production of sodium taurine deoxycholate according to claim 1, characterized in that, The above-described method for mass production of sodium taurine deoxycholate can produce more than 1 kg of sodium taurine deoxycholate per batch.

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