Novel processes for preparing diaminopyrimidine derivative or acid addition salt thereof
A novel method for preparing diaminopyrimidine derivatives and their acid addition salts addresses the instability and impurity issues of previous methods by using a substituted phenylguanidine derivative and (S)-3-acetamidopyrrolidine, achieving high yield, purity, and stability suitable for industrial production.
Patent Information
- Application Number
- JP2025148068
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2018-05-18
- Filing Date
- 2025-09-08
- Publication Date
- 2025-11-26
AI Technical Summary
Existing methods for preparing diaminopyrimidine derivatives and their acid addition salts, such as those described in WO2012/115480, suffer from issues like the use of unstable intermediates, low yields, structural isomer impurities, and poor stability, making them unsuitable for industrial mass production and leading to the formation of unwanted impurities.
A novel method involving the formation of a pyrimidine ring from a substituted phenylguanidine derivative or its salt, followed by activation and reaction with (S)-3-acetamidopyrrolidine, avoids the use of unstable intermediates and includes the production of a novel crystalline form of the hydrochloride salt through controlled conditions, enhancing stability and suitability for industrial production.
The new method achieves high yield and purity of the diaminopyrimidine derivative or its acid addition salt, suitable for industrial mass production, and the novel crystalline forms exhibit superior stability compared to amorphous forms, ensuring minimal impurities and improved product integrity.
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Abstract
Description
[Technical Field]
[0001] The present invention relates to a novel method for preparing a diaminopyrimidine derivative or an acid addition salt thereof having activity as a 5-HT4 receptor agonist, and also to a novel crystalline form of the hydrochloride salt of a diaminopyrimidine derivative and a method for preparing the same. [Background technology]
[0002] The diaminopyrimidine derivative of the following chemical formula 1 has the chemical name (S)—N-(1-(2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl)pyrrolidin-3-yl)acetamide. The diaminopyrimidine derivative of the following chemical formula 1 or a pharmaceutically acceptable salt (e.g., hydrochloride) thereof acts as a 5-HT4 receptor agonist and can therefore be usefully applied in the prevention or treatment of gastrointestinal motility disorders, which are gastrointestinal diseases, such as gastroesophageal reflux disease (GERD), constipation, irritable bowel syndrome (IBS), dyspepsia, postoperative ileus, delayed gastric emptying, gastroparesis, intestinal pseudo-obstruction, drug-induced delayed transit, or diabetic gastroparesis (WO2012 / 115480). [ka]
[0003] A method for preparing the compound of Chemical Formula 1 is disclosed in WO2012 / 115480. Specifically, as shown in the following Reaction Scheme 1, the method for preparing the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof includes the steps of reacting a compound of Chemical Formula (I) with POCl to prepare a compound of Chemical Formula (II), reacting the compound of Chemical Formula (II) with a compound of Chemical Formula (III) in the presence of a base to prepare a compound of Chemical Formula (IV), reacting the compound of Chemical Formula (IV) with a compound of Chemical Formula (V) to prepare a compound of Chemical Formula 1, and optionally converting the compound of Chemical Formula 1 into a pharmaceutically acceptable salt thereof. [ka]
[0004] In the preparation method of Reaction Scheme 1, the compound of formula (II) is prepared by reacting with an excess amount of POCl3 at a high temperature of 100°C or higher, resulting in the generation of a large amount of impurities. Therefore, to remove the impurities, the reaction requires a purification step using column chromatography, which is not suitable for industrial mass production. In addition, the compound of formula (II) is obtained in the form of an oil and is easily decomposed at room temperature, resulting in a stability problem, i.e., poor stability.
[0005] In particular, because the compound of formula (II) has two reactive sites (i.e., positions 2 and 4 of the pyrimidine ring), a large amount of a structural isomer impurity in which the compound of formula (III) is bonded to position 2 of the pyrimidine ring is produced. Therefore, the reaction of the compound of formula (II) with the compound of formula (III) shows a very low yield of about 55-60%, which poses the problem of the structural isomer impurity remaining in the final product (i.e., the compound of formula 1).
[0006] Furthermore, in the reaction between the compound of formula (IV) and the compound of formula (V), the compound of formula (V) is known to be a genotoxic substance. Therefore, in order to avoid the compound of formula (V) remaining in the final product (i.e., the compound of formula 1), a purification step using column chromatography, which is not suitable for industrial mass production, is required.
[0007] WO2012 / 115480 also discloses a process for converting a compound of Chemical Formula 1 into its hydrochloride salt. The conversion process includes reacting the compound of Chemical Formula 1 with hydrochloric acid in ethyl acetate, stirring at room temperature for 1 hour, and then filtering the product. The inventors discovered that the hydrochloride salt of the compound of Chemical Formula 1 obtained by this process is amorphous and has the problem of producing a large amount of deacetylated impurities when stored at room temperature. Summary of the Invention [Problem to be solved by the invention]
[0008] The present inventors have conducted extensive research to develop a novel method for preparing the compound of Formula 1 or its acid addition salt, which can solve the problems of the prior art methods for preparing the compound of Formula 1 or its acid addition salt. In particular, the present inventors have conducted extensive research to develop a novel method for avoiding the use of the compound of Formula (II), i.e., an intermediate that is unstable, difficult to handle, and forms structural isomer impurities. As a result, the present inventors have developed a novel method comprising the steps of forming a pyrimidine ring from a substituted phenylguanidine derivative or its salt, activating the pyrimidine ring to prepare a novel intermediate, and reacting the novel intermediate with (S)-3-acetamidopyrrolidine. The method has been found to solve the problems of the prior art methods and to be suitable for industrial mass production.
[0009] The present inventors have also isolated a novel crystalline form of the hydrochloride salt of the compound of Formula 1 and found that said crystalline form exhibits significantly superior stability compared to the amorphous form of the prior art.
[0010] It is therefore an object of the present invention to provide a process for preparing a diaminopyrimidine derivative or an acid addition salt thereof, which process involves the use of a novel intermediate.
[0011] Another object of the present invention is to provide the novel intermediate and a method for producing the same.
[0012] Yet another object of the present invention is to provide a crystalline form of the hydrochloride salt of the compound of Formula 1 and a method for preparing the same. [Means for solving the problem]
[0013] According to one aspect of the present invention, a compound of Formula 1: [ka] or an acid addition salt thereof, comprising: (a) Chemical formula 2: [ka] (In the formula, R 1 is a C1-C3 alkyl group; or a phenyl group optionally substituted with one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, and cyano). or a salt thereof with (S)-3-acetamidopyrrolidine to produce a compound of formula 1; and Optionally, (b) reacting the compound of Formula 1 with an acid to prepare an acid addition salt of the compound of Formula 1. A method is provided that includes:
[0014] According to another aspect of the present invention, a compound of Formula 2: [ka] (In the formula, R 1 is the same as defined above) or a salt thereof.
[0015] According to yet another aspect of the present invention, there is provided a method for producing a compound of formula 2 or a salt thereof, comprising the steps of: (i) Chemical formula 5: [ka] or a salt thereof with methyl 3-oxohexanoate to form a compound of formula 4: [ka] a process for producing a compound of formula (I); (ii) the compound of formula 4 and R 1 with a sulfonyl halide to produce a compound of formula 2; and Optionally, (iii) converting the compound of formula 2 into a salt thereof. A method is provided that includes:
[0016] According to yet another aspect of the present invention, there is provided a compound of formula 1: [ka] Crystalline Form A of the hydrochloride salt of the compound of formula (I) is provided.
[0017] According to yet another aspect of the present invention, there is provided a method for preparing crystalline form A of the hydrochloride salt of compound of formula 1, comprising the steps of: (p) adding hydrochloric acid to the compound of formula 1 in an organic solvent and refluxing the resulting reaction mixture; (q) cooling the reaction mixture obtained in step (p) to form a precipitate; and (r) isolating the precipitate from step (q). A method is provided which includes:
[0018] According to yet another aspect of the present invention, there is provided a compound of formula 1: [ka] Crystalline Form B of the hydrochloride salt of the compound of formula (I) is provided.
[0019] According to yet another aspect of the present invention, there is provided a method for preparing crystalline form B of the hydrochloride salt of compound of formula 1, comprising the steps of: A method is provided that includes the step of exposing crystalline Form A of the hydrochloride salt of the compound of Formula 1 to a condition of 40% or greater relative humidity to adjust the moisture content to about 3.5% or greater. [Effects of the Invention]
[0020] The method of the present invention is carried out using a novel intermediate, i.e., the compound of formula 2 or its salt, obtained from a substituted phenylguanidine derivative or its salt. Therefore, the method of the present invention can avoid the use of the compound of formula (II), an intermediate that is unstable, difficult to handle, and forms structural isomer impurities. Furthermore, the method of the present invention can provide the compound of formula 1 or its acid addition salt in high yield and purity, making it suitable for industrial mass production. Furthermore, the novel crystalline forms of the hydrochloride salt of the compound of formula 1, i.e., crystalline form A and crystalline form B, exhibit significantly superior stability compared to the amorphous form of the prior art. [Brief explanation of the drawings]
[0021] [Figure 1] 1 shows the XRPD spectrum of crystalline form A of the hydrochloride salt of the compound of formula 1. [Figure 2] 1 shows the XRPD spectrum of crystalline form B of the hydrochloride salt of the compound of formula 1. DETAILED DESCRIPTION OF THE INVENTION
[0022] The present invention relates to a compound of formula 1: [ka] or an acid addition salt thereof, comprising: (a) Chemical formula 2: [ka] (In the formula, R 1 is a C1-C3 alkyl group; or a phenyl group optionally substituted with one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, and cyano). or a salt thereof with (S)-3-acetamidopyrrolidine to produce a compound of formula 1; and Optionally, (b) reacting the compound of Formula 1 with an acid to prepare an acid addition salt of the compound of Formula 1. The present invention provides a method comprising:
[0023] In the production method of the present invention, one of the reactants, (S)-3-acetamidopyrrolidine, is a known compound and commercially available. Alternatively, (S)-3-acetamidopyrrolidine may be formed by deprotecting the amino-protecting group from a commercially available compound of the following chemical formula 3: [ka] (In the formula, R 2 is an amino protecting group such as t-butoxycarbonyl.
[0024] The deprotection may be carried out using one or more acids selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, formic acid, sulfonic acid, p-toluenesulfonic acid, and methanesulfonic acid, preferably p-toluenesulfonic acid or hydrochloric acid. The deprotection may be carried out in one or more solvents selected from the group consisting of dichloromethane, dichloroethane, N,N-dimethylformamide, dimethylacetamide, dimethyl sulfoxide, tetrahydrofuran, ethyl acetate, and toluene, preferably toluene. The deprotection may be carried out at a temperature ranging from 0°C to 40°C, preferably from 10°C to 30°C.
[0025] In one embodiment, the deprotection and the reaction of the resulting (S)-3-acetamidopyrrolidine with the compound of Chemical Formula 2 or a salt thereof may be carried out in a one-pot reaction. That is, in the production method of the present invention, step (a) may include: (a1) deprotecting the compound of Chemical Formula 3 to form (S)-3-acetamidopyrrolidine; and (a2) adding the compound of Chemical Formula 2 or a salt thereof to the reaction mixture of step (a1) to carry out the reaction. The reaction of the compound of Chemical Formula 2 or a salt thereof with (S)-3-acetamidopyrrolidine may be carried out in the presence of a base. The base may be one or more selected from the group consisting of potassium tert-butoxide, sodium hydroxide, potassium hydroxide, sodium hydride, sodium carbonate, potassium carbonate, potassium phosphate (including monopotassium phosphate, dipotassium phosphate, and tripotassium phosphate), sodium phosphate (including monosodium phosphate, disodium phosphate, and trisodium phosphate), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), 1,4-diazabicyclo[2.2.2]octane (DABCO), 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), pyridine, triethylamine, diisopropylamine, and diisopropylethylamine. Preferably, the base may be triethylamine or diisopropylethylamine. More preferably, the base may be diisopropylethylamine. The reaction of the compound of formula 2 or a salt thereof with (S)-3-acetamidopyrrolidine may be carried out at a temperature ranging from 40°C to 130°C, preferably from 80°C to 90°C.
[0026] In the production method of the present invention, the acid used in step (b) may be a conventional inorganic or organic acid suitable for forming the desired acid addition salt. Preferably, the acid used in step (b) may be hydrochloric acid. When hydrochloric acid is used in step (b), the product is obtained in the form of the hydrochloride salt of the compound of Formula 1.
[0027] In the production method of the present invention, the compound of Chemical Formula 2 or a salt thereof is (i) Chemical formula 5: [ka] or a salt thereof with methyl 3-oxohexanoate to form a compound of formula 4: [ka] a process for producing a compound of formula (I); (ii) the compound of formula 4 and R 1 with a sulfonyl halide to produce a compound of formula 2; and Optionally, (iii) converting the compound of formula 2 into a salt thereof. The film may be manufactured or formed by a manufacturing method including:
[0028] In the reaction of step (i), the compound of Chemical Formula 5 may be prepared by reacting 4-amino-2-nitroaniline with cyanamide under acidic conditions according to known methods, such as those described in U.S. Pat. No. 6,562,854 B2. The salt of the compound of Chemical Formula 5 may be in the form of 1-(4-amino-3-nitrophenyl)guanidine hemicarbonate. The reaction of step (i), i.e., the cyclization reaction, may be carried out in a C1-C5 alcohol, preferably n-butanol. The cyclization reaction may be carried out at a temperature ranging from 50°C to 130°C, preferably from 90°C to 120°C.
[0029] The reaction of step (ii), i.e., the reaction of the compound of formula 4 with R 1The reaction with the -sulfonyl halide may be carried out in the presence of a base. The base may be one or more selected from the group consisting of potassium tert-butoxide, sodium hydroxide, potassium hydroxide, sodium hydride, sodium carbonate, potassium carbonate, potassium phosphate (including potassium monophosphate, potassium diphosphate, and potassium triphosphate), sodium phosphate (including sodium monophosphate, sodium diphosphate, and sodium triphosphate), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), 1,4-diazabicyclo[2.2.2]octane (DABCO), 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), pyridine, triethylamine, diisopropylamine, and diisopropylethylamine. Preferably, the base may be triethylamine. The compound of Formula 4 and R 1 The reaction with the -sulfonyl halide may be carried out in the presence or absence of a solvent. When the reaction is carried out in the presence of a solvent, the solvent may be one or more selected from the group consisting of acetone, methyl ethyl ketone, dichloromethane, dichloroethane, N,N-dimethylformamide, dimethylacetamide, dimethyl sulfoxide, tetrahydrofuran, C1-C5 alcohols, ethyl acetate, and toluene. In addition, the reaction of the compound of formula 4 with R 1 The reaction with the -sulfonyl halide may be carried out at a temperature ranging from 0° C. to 60° C. If desired, the compound of formula 4 may be reacted with R 1 The compound of formula 2 obtained from the reaction with the -sulfonyl halide may be further converted into a salt form (e.g., an acid addition salt form) by reacting with an acid such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, formic acid, or sulfonic acid. The resulting compound of formula 2 or a salt thereof may be isolated. Alternatively, the resulting compound of formula 2 or a salt thereof may be directly used in the next reaction, i.e., the reaction with (S)-3-acetamidopyrrolidine, without isolation. In one embodiment, the reaction mixture of step (ii) may be directly used in the reaction of step (a) without isolating the compound of formula 2 or a salt thereof.
[0030] An example of the overall reaction process of the present invention is shown in Reaction Scheme 2 below. [ka] (In Reaction Scheme 2, R 1 is the same as defined above.)
[0031] The present invention also provides a compound of formula 2, or a salt thereof, useful for preparing a compound of formula 1, or an acid addition salt thereof. [ka] (In the formula, R 1 is the same as defined above.)
[0032] The present invention also provides a method for producing a compound of Chemical Formula 2 or a salt thereof, namely, a method for producing a compound of Chemical Formula 2 or a salt thereof, comprising: (i) Chemical formula 5: [ka] or a salt thereof with methyl 3-oxohexanoate to form a compound of formula 4: [ka] a process for producing a compound of formula (I); (ii) the compound of formula 4 and R 1 with a sulfonyl halide to produce a compound of formula 2; and Optionally, (iii) converting the compound of formula 2 into a salt thereof. The present invention provides a method comprising: The reactions in steps (i) and (ii) are as described above.
[0033] The present invention also provides a novel crystalline form of the hydrochloride salt of the compound of Formula 1, which has excellent stability.
[0034] In one aspect, the present invention provides a compound of formula 1: [ka] The present invention provides crystalline form A of the hydrochloride salt of the compound of formula (I).
[0035] Preferably, crystalline Form A of the hydrochloride salt of the compound of Chemical Formula 1 may have the XRPD pattern of Figure 1. Also, crystalline Form A of the hydrochloride salt of the compound of Chemical Formula 1 may have a differential scanning calorimetry (DSC) thermogram showing an endothermic peak at 229°C to 235°C, preferably at 232±2°C. The onset temperature may be 230±2°C.
[0036] The present invention also encompasses within its scope a method for preparing crystalline Form A of the hydrochloride salt of the compound of Chemical Formula 1. Specifically, the present invention provides a method for preparing crystalline Form A of the hydrochloride salt of the compound of Chemical Formula 1, comprising the steps of: (p) adding hydrochloric acid to the compound of Chemical Formula 1 in an organic solvent and refluxing the resulting reaction mixture; (q) cooling the reaction mixture obtained in step (p) (usually to room temperature) to form a precipitate; and (r) isolating the precipitate obtained in step (q). The organic solvent used in step (p) may be one or more selected from the group consisting of acetone, methyl ethyl ketone, ethyl acetate, acetonitrile, tetrahydrofuran, heptane, and C1-C5 alcohols. For example, a mixed solvent of acetone and isopropyl alcohol may be used as the organic solvent. The volume ratio of acetone to isopropyl alcohol in the mixed solvent may be in the range of 200-280:100, preferably about 250:100. The refluxing may be carried out at a temperature in the range of 50°C to 120°C, preferably 55°C to 70°C.
[0037] In another aspect, the present invention provides a compound of formula 1: [ka] The present invention provides crystalline form B of the hydrochloride salt of the compound of formula (I).
[0038] Preferably, crystalline form B of the hydrochloride salt of the compound of Chemical Formula 1 may have the XRPD pattern of Figure 2. Also, crystalline form B of the hydrochloride salt of the compound of Chemical Formula 1 may have a differential scanning calorimetry (DSC) thermogram showing an endothermic peak at 229°C to 235°C, preferably at 232±2°C. The onset temperature may be 230±2°C.
[0039] The present invention also includes within its scope a method for preparing crystalline Form B of the hydrochloride salt of compound of Chemical Formula 1. Specifically, the present invention provides a method for preparing crystalline Form B of the hydrochloride salt of compound of Chemical Formula 1, which comprises exposing crystalline Form A of the hydrochloride salt of compound of Chemical Formula 1 to a relative humidity of 40% or higher to adjust the water content to about 3.5% or higher.
[0040] The present invention will be described in more detail below with reference to examples. However, these examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention. [Example]
[0041] Preparation Example 1: 1-(4-amino-3-nitrophenyl)guanidine hemicarbonate A mixture of 4-amino-2-nitroaniline (100.0 g, 0.653 mol), purified water (200 ml), concentrated hydrochloric acid (61.2 g, 0.588 mol), and cyanamide (35.7 g, 0.849 mol) was stirred at 80-90°C for 3 hours. A solution of sodium carbonate (41.5 g, 0.392 mol) in purified water (500 ml) was slowly added to the reaction mixture at approximately 65°C. The reaction mixture was stirred for 2 hours, cooled to approximately 10°C, and then filtered. The resulting solid was dried under vacuum to give 112.5 g of the title compound (yield: 76.2%). 1 H-NMR(600MHz, DMSO) δ 9.86(br, 1H), 7.80(s, 1H), 7.66(s, 2H), 7.56(s, 2H), 7.29(s, 1H), 7.15(s, 1H)
[0042] Example 1: 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-ol A mixture of 1-(4-amino-3-nitrophenyl)guanidine hemicarbonate (110.0 g, 0.486 mol), methyl 3-oxohexanoate (91.1 g, 0.632 mol), and n-butanol (330 ml) was stirred at 95° C. for 15 hours. The reaction mixture was cooled to room temperature and then filtered. The resulting solid was dried under reduced pressure to give 111.3 g of the title compound (yield: 79.1%, HPLC purity: 98.2%). 1 H-NMR(400MHz, DMSO) δ 10.82(br, 1H), 8.98(br, 1H), 8.57(s, 1H), 7.49(d, 1H), 7.30(s, 2H), 7.02(d, 1H), 5.62(s, 1H), 2.35(t, 2H), 1.72-1.63(m, 2H), 0.91(t, 3H)
[0043] Example 2: 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl 4-methylbenzenesulfonate To a solution of 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-ol (106.5 g, 0.368 mol) and p-toluenesulfonyl chloride (64.27 g, 0.337 mol) in acetone (700 ml) was added triethylamine (37.2 g, 0.368 mol). The reaction mixture was stirred at room temperature for 3 hours and then filtered. Toluene (1,065 ml) was added to the obtained filtrate, which was then concentrated to 430 ml to remove acetone. The obtained solution was directly used in the preparation of the compound of formula 1. To identify the obtained product, a portion of the solution was concentrated under reduced pressure. 1 The H-NMR spectrum data is as follows: 1 H-NMR(400MHz, DMSO) δ 9.73(s, 1H), 8.33(br, 1H), 7.90(d, 2H), 7.56(d, 1H), 7.43(d, 2H), 7.32(s, 2H), 6.96(d, 1H), 6.44(s, 1H), 2.59(t, 2H), 2.40(s, 3H), 1.74-1.65(m, 2H), 1.18(t, 3H)
[0044] Example 3: 4-Methylbenzenesulfonate salt of 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl 4-methylbenzenesulfonate To a solution of 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-ol (20.0 g, 0.069 mol) and p-toluenesulfonyl chloride (14.5 g, 0.076 mol) in acetone (170 ml) was added triethylamine (8.4 g, 0.083 mol). The reaction mixture was stirred at room temperature for 3 hours and then filtered. To the obtained filtrate was added p-toluenesulfonic acid (14.5 g, 0.076 mol). The mixture was stirred at room temperature for 2 hours and then filtered. The obtained solid was dried under reduced pressure to give 37.9 g of the title compound (yield: 89.1%). 1H-NMR(400MHz, CDCl3) δ 11.70(s, 1H), 8.31(s, 1H), 7.81(d, 2H), 7.66(d, 2H), 7.56(d, 1H), 7.43(d, 2H), 7.32(s, 2H), 6.96(d, 1H), 6.44(s, 1H), 2.59(t, 2H), 2.40(s, 3H), 1.74-1.65(m, 2H), 1.18(t, 3H)
[0045] Example 4: 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl methanesulfonate To a solution of 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-ol (18.0 g, 0.062 mol) and methanesulfonyl chloride (7.8 g, 0.068 mol) in N,N-dimethylformamide (54 ml) was added triethylamine (7.6 g, 0.075 mol). The reaction mixture was stirred at room temperature for 3 hours. Purified water (180 ml) and toluene (54 ml) were added to the reaction mixture, which was then filtered. The resulting solid was dried under reduced pressure to give 18.0 g of the title compound (yield: 79.0%). 1 H-NMR(400MHz, DMSO) δ 10.62(br, 1H), 8.76(br, 1H), 8.54(s, 1H), 7.48(d, 1H), 7.31(s, 2H), 6.99(d, 1H), 5.66(s, 1H), 3.72(s, 3H), 2.34(t, 2H), 1.70-1.64(m, 2H), 0.91(t, 3H)
[0046] Example 5: 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl 4-bromobenzenesulfonate To a solution of 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-ol (10.0 g, 0.035 mol) and 4-bromobenzenesulfonyl chloride (9.7 g, 0.038 mol) in N,N-dimethylformamide (30 ml) was added triethylamine (4.2 g, 0.041 mol). The reaction mixture was stirred at room temperature for 3 hours. Purified water (180 ml) and toluene (54 ml) were added to the reaction mixture. The extracted organic layer was used directly for the preparation of the compound of formula 1. To identify the obtained product, a portion of the organic layer was concentrated under reduced pressure. 1 The H-NMR spectrum data is as follows: 1 H-NMR(400MHz, DMSO) δ 9.72(s, 1H), 7.96(br, 2H), 7.83(d, 2H), 7.56(d, 1H), 7.27(s, 2H), 6.98(d, 1H), 6.47(s, 1H), 2.59(t, 2H), 1.74-1.65(m, 2H), 0.90(t, 3H)
[0047] Example 6: (S)—N-(1-(2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl)pyrrolidin-3-yl)acetamide To a mixture of tert-butyl (S)-3-acetamidopyrrolidine-1-carboxylate (70.0 g, 0.307 mol) and toluene (700 ml), p-toluenesulfonic acid monohydrate (116.7 g, 0.613 mol) was added. The reaction mixture was stirred at room temperature for 6 hours. To the reaction mixture, a toluene solution of 2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl 4-methylbenzenesulfonate obtained in Example 2 and diisopropylethylamine (198.1 g, 1.533 mol) were added. The reaction mixture was stirred at 80-90°C for 5 hours and then concentrated under reduced pressure. To the resulting residue, methanol (210 ml) and 50% sodium bicarbonate solution (70 ml) were added. The resulting mixture was stirred at room temperature for 2 hours, and then distilled water (1,050 ml) was added dropwise. The reaction mixture was filtered, and the resulting solid was dried under reduced pressure to give 105.5 g of the title compound (yield: 86.0%, HPLC purity: 99.1%). 1H-NMR(400MHz, DMSO) δ 8.99(s, 2H), 8.17(s, 1H), 7.58(d, 1H), 7.16(s, 2H), 6.93(d, 1H), 5.79(s, 1H), 4.35(m, 1H), 3.70-3.35(m, 6H), 2.40(t, 2H), 1.82(s, 3H), 1.71-1.65(m, 2H), 0.92(t, 3H)
[0048] Example 7: (S)—N-(1-(2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl)pyrrolidin-3-yl)acetamide hydrochloride (crystalline form A) To a mixture of (S)—N-(1-(2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl)pyrrolidin-3-yl)acetamide (10.0 g, 0.025 mol), acetone (250 ml), and isopropyl alcohol (100 ml) was added concentrated hydrochloric acid (2.6 g, 0.025 mol) at 50° C. The reaction mixture was refluxed and stirred for 20 hours, cooled to room temperature, and then filtered. The resulting solid was dried under reduced pressure at approximately 60° C. to give 9.8 g of the title compound (yield: 90.0%, HPLC purity: 99.9%). 1 H-NMR(600MHz) δ 8.56(d, 1H), 7.37-7.34(m, 1H), 6.96-6.93(m, 1H), 6.09(d, 1H), 4.53-4.47(d, 1H), 3.91-3.43(m, 4H), 2.62-2.58(m, 2H), 2.36-2.28(m, 1H), 2.12-2.05(m, 1H), 1.98(s, 3H), 1.78-1.74(m, 2H), 1.06-1.03(t, 3H)
[0049] X-ray powder diffraction (XRPD) analysis of the resulting product was performed. X-ray powder diffraction (XRPD) patterns were collected on a Bruker D8 Advance diffractometer operated at 40 kV and 40 mA using Cu-Kα radiation (λ = 1.5406 Å). XRPD patterns were collected by scanning from 2θ = 4° to 40° in 0.02° increments at a scan rate of 0.1 s / step (divergence slit: 0.3 and anti-scatter slit: 0.3). The resulting XRPD spectrum is shown in Figure 1. Figure 1 shows characteristic peaks at 2θ = 7.4°, 9.1°, 12.0°, 12.5°, 13.5°, 14.1°, 15.9°, 16.8°, 18.3°, 19.1°, 24.6°, 25.3°, and 26.8° ± 0.2°.
[0050] The resulting product was analyzed by differential scanning calorimetry (DSC). Differential scanning calorimetry (DSC) was performed using a Mettler-Toledo DSC 1 STAR differential scanning calorimeter. The analytical conditions were as follows: the sample was placed in an aluminum pan; the pan was covered with an aluminum lid (with a pinhole); the pan was purged with 99% nitrogen gas (gas flow rate: 50 mL / min); the starting temperature was 30°C; the ending temperature was 300°C; and the heating rate was 10°C / min. An endothermic peak was observed between 229°C and 235°C. The onset temperature was 230.4°C, and the maximum peak was observed at 233.0°C.
[0051] Example 8: (S)—N-(1-(2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl)pyrrolidin-3-yl)acetamide hydrochloride (crystalline form B) Crystalline Form A of (S)—N-(1-(2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl)pyrrolidin-3-yl)acetamide hydrochloride prepared in Example 7 was exposed to 40% relative humidity in a humid chamber for at least 5 hours to give the title compound. 1H-NMR(600MHz) δ 8.56(d, 1H), 7.37-7.34(m, 1H), 6.96-6.93(m, 1H), 6.09(d, 1H), 4.53-4.47(d, 1H), 3.91-3.43(m, 4H), 2.62-2.58(m, 2H), 2.36-2.28(m, 1H), 2.12-2.05(m, 1H), 1.98(s, 3H), 1.78-1.74(m, 2H), 1.06-1.03(t, 3H)
[0052] The resulting product was analyzed by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC) using the same methods as in Example 7. The resulting XRPD spectrum is shown in Figure 2. Figure 2 shows characteristic peaks at 2θ = 7.3°, 8.8°, 11.8°, 12.5°, 13.2°, 13.6°, 14.5°, 17.7°, 18.6°, 19.6°, 22.0°, 24.9°, 25.2°, and 25.9° ± 0.2°. Differential scanning calorimetry (DSC) analysis revealed an endothermic peak between 229°C and 235°C. The onset temperature was 229.4°C, and the maximum peak was observed at 232.2°C.
[0053] Test Example 1: Stability test (S)—N-(1-(2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl)pyrrolidin-3-yl)acetamide hydrochloride (amorphous, test compound 1) prepared according to WO2012 / 115480 and crystalline form A of (S)—N-(1-(2-((4-amino-3-nitrophenyl)amino)-6-propylpyrimidin-4-yl)pyrrolidin-3-yl)acetamide hydrochloride (test compound 2) prepared in Example 7 were stored under conditions of 60° C. and 75% RH for 2 weeks. The amounts of each test compound and deacetylated impurity were analyzed by HPLC to determine their HPLC purity. The results are shown in Table 1 below.
[0054] [Table 1]
[0055] As shown in Table 1 above, crystalline form A according to the present invention not only exhibited high purity from the beginning, but also showed no significant changes in HPLC purity and the amount of deacetylated impurity under conditions of 60°C and 75% RH. In contrast, the amorphous form according to the prior art not only exhibited a large amount of deacetylated impurity from the beginning, but also showed a significant decrease in HPLC purity and a significant increase in the amount of deacetylated impurity under conditions of 60°C and 75% RH. Therefore, the above results confirm that the crystalline form according to the present invention has excellent stability.
Claims
1. 1, having an XRPD pattern with peaks in degree of two-theta at 7.4, 9.1, 12.0, 12.5, 13.5, 14.1, 15.9, 16.8, 18.3, 19.1, 24.6, 25.3, and 26.8°±0.2°; 【Chemistry 1】 Crystal A of the hydrochloride salt of the compound of formula (I).
2. 2. Crystalline hydrochloride salt A of the compound of formula 1 according to claim 1, which has a differential scanning calorimetry (DSC) thermogram showing an endothermic peak at 229°C to 235°C.
3. A method for preparing Crystal A of the hydrochloride salt of the compound of formula 1 according to claim 1 or 2, comprising the steps of: (p) adding hydrochloric acid to the compound of Formula 1 in an organic solvent and refluxing the resulting reaction mixture; (q) cooling the reaction mixture obtained in step (p) to form a precipitate; and (r) isolating the precipitate of step (q). A method comprising:
4. 2. Compound (I) having an XRPD pattern with peaks in degree of two-theta at 7.3, 8.8, 11.8, 12.5, 13.2, 13.6, 14.5, 17.7, 18.6, 19.6, 22.0, 24.9, 25.2, and 25.9°±0.2°, chemical formula 1: 【Chemistry 2】 Crystal B of the hydrochloride salt of the compound of formula (I).
5. 5. Crystalline hydrochloride salt B of the compound of formula 1 according to claim 4, having a differential scanning calorimetry (DSC) thermogram showing an endothermic peak at 229°C to 235°C.
6. A method for producing crystalline B of the hydrochloride salt of the compound of chemical formula 1 according to claim 4 or 5, comprising the step of exposing crystalline A of the hydrochloride salt of the compound of chemical formula 1 according to claim 1 to a condition of a relative humidity of 40% or more to adjust the water content to 3.5% or more.