Novel intermediate and method for preparing dimer using same

Abstract

The present disclosure relates to a method for preparing high-purity H-AEEA-AEEA-OH (17-amino-10-oxo -3,6,12,15-tetraoxa-9-azaheptadecanoic acid) and a novel intermediate used in the method.

Description

A novel intermediate and a method for producing a dimer using the same

[0001] The present invention relates to a method for producing high-purity H-AEEA-AEEA-OH (17-Amino-10-oxo-3,6,12,15-tetraoxa-9-azaheptadecanoic acid) and a novel intermediate used in the method.

[0002] Semaglutide, approved by the Food and Drug Administration (FDA) in 2017, is a glucagon-like peptide-1 (GLP-1) drug known to lower blood sugar levels in patients with type 2 diabetes by restoring insulin production. Furthermore, semaglutide is known to promote insulin secretion while suppressing glucagon secretion, thereby increasing satiety and reducing appetite, thereby aiding weight loss in patients taking the drug.

[0003] Additionally, tirzepatide, approved by the FDA in 2022, is known to act as a dual agonist at the glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptors. Specifically, tirzepatide exhibits the effects of both insulinotropic peptides, allowing it to simultaneously activate the dual action mechanisms of GIP and GLP-1.

[0004] Semaglutide and terzepatide, described above, are synthesized to include side chains to extend their half-life and provide prolonged effects in the body. Among these side chains, the most commonly used is H-AEEA-AEEA-OH, represented by the following chemical formula 4.

[0005] [Chemical Formula 4]

[0006]

[0007] The above H-AEEA-AEEA-OH has the effect of easily dissolving the bound drug, thereby increasing the mobility of the drug. In addition, the H-AEEA-AEEA-OH has the effect of inhibiting the filtration of the bound drug in the kidney and protecting the drug from the body's immune system.

[0008] The conventional method for manufacturing H-AEEA-AEEA-OH has been to reduce the reactivity of the amine group by introducing an amine group protecting group into H-AEEA-OH (2-(2-(2-Aminoethoxy)ethoxy)acetic acid), increase the reactivity by ester bonding the carboxyl group with an activator such as hydrosuccinimide, hydroxybenzotriazole, or imidazole, and then manufacture H-AEEA-AEEA-OH through a polymerization reaction. However, the conventional manufacturing method had a disadvantage in that the activator was easily decomposed during the polymerization reaction. Due to this, a large amount of impurities that did not participate in the polymerization reaction were generated, which resulted in a problem in that the purity of the product H-AEEA-AEEA-OH was low.

[0009] At this time, the purity of H-AEEA-AEEA-OH used as a side chain in the drug has a significant impact on the purity of the entire drug containing it, which may reduce the economic feasibility of the entire drug manufacturing process. Therefore, there is a need to develop a method for manufacturing H-AEEA-AEEA-OH capable of producing high-purity H-AEEA-AEEA-OH.

[0010] [Prior Art Literature]

[0011] (Patent Document 1) Chinese Publication Patent No. 119462413.

[0012] The present invention provides a method for producing high-purity H-AEEA-AEEA-OH (17-Amino-10-oxo-3,6,12,15-tetraoxa-9-azaheptadecanoic acid) and a novel intermediate used in the method.

[0013] However, the problems that the present invention seeks to solve are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0014] A first aspect of the present invention provides a compound represented by the following chemical formula 1:

[0015] [Chemical Formula 1]

[0016] ;

[0017] In the above chemical formula 1, Cbz is a benzyloxycarbonyl group, and the R 1 Inland R 4 are, each independently, hydrogen, halogen, linear or branched C 1-5 Alkyl group, and linear or branched C 1-5 It is selected from alkoxy groups.

[0018] The second aspect of the present invention provides a method for preparing a compound according to the first aspect, comprising the step of reacting H-AEEA-OH(2-(2-(2-Aminoethoxy)ethoxy)acetic acid) and benzyl chloroformate.

[0019] The third aspect of the present invention provides a method for preparing H-AEEA-AEEA-OH, comprising the steps of: (a) reacting a compound represented by the following chemical formula 1 with H-AEEA-OH (2-(2-(2-Aminoethoxy)ethoxy)acetic acid) to obtain a first reaction intermediate represented by the following chemical formula 2; and (b) removing a Cbz protecting group from the first reaction intermediate to obtain H-AEEA-AEEA-OH.

[0020] [Chemical Formula 1]

[0021] ;

[0022] [Chemical Formula 2]

[0023] ;

[0024] In the above chemical formulas 1 and 2, Cbz is a benzyloxycarbonyl group, and R 1 Inland R 4 are, each independently, hydrogen, halogen, linear or branched C 1-5 Alkyl group, and linear or branched C 1-5 It is selected from alkoxy groups.

[0025] The compound according to the embodiments of the present invention, represented by Chemical Formula 1, exists in a stable state and has the characteristic of generating less impurities (Cbz-AEEA-OH) during the reaction. In addition, the compound may exist in a solid state, making it easy to measure and store.

[0026] According to the method for producing H-AEEA-AEEA-OH according to the embodiments of the present disclosure, H-AEEA-AEEA-OH can be produced in a high yield. The method for producing H-AEEA-AEEA-OH according to the embodiments of the present disclosure can produce H-AEEA-AEEA-OH under mild conditions.

[0027] Figure 1 is a reaction scheme of a method for producing H-AEEA-AEEA-OH (17-Amino-10-oxo-3,6,12,15-tetraoxa-9-azaheptadecanoic acid) according to one embodiment of the present invention.

[0028] Figure 2 shows the results of HPLC (High-performance liquid chromatography) analysis of Cbz-AEEA-OH synthesized according to Example 1 of the present invention.

[0029] Figure 3 is an NMR (Nuclear Magnetic Resonance Spectroscopy) spectrum of Cbz-AEEA-Bt synthesized according to Example 1 of the present invention.

[0030] Figure 4 shows the HPLC analysis results of Cbz-AEEA-AEEA-OH synthesized according to Example 2 of the present invention.

[0031] Figure 5 shows the HLPC analysis results of H-AEEA-AEEA-OH synthesized according to Example 4 of the present invention.

[0032] Figure 6 shows the HPLC analysis results of Cbz-AEEA-OH synthesized according to Comparative Example 1 of the present invention.

[0033] Figure 7 shows the HPLC analysis results of Cbz-AEEA-AEEA-OH synthesized according to Comparative Example 2 of the present invention.

[0034] Hereinafter, with reference to the attached drawings, implementation examples and embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. However, the present invention may be implemented in various different forms and is not limited to the implementation examples and embodiments described herein. In addition, in the drawings, parts irrelevant to the description have been omitted to clearly explain the present invention, and similar parts have been designated with similar drawing reference numerals throughout the specification.

[0035] Throughout this specification, when a part is said to be "connected" to another part, this includes not only cases where it is "directly connected" but also cases where it is "electrically connected" with another element in between.

[0036] Throughout this specification, when it is said that an element is "on" another element, this includes not only cases where the element is in contact with the other element, but also cases where another element exists between the two elements.

[0037] Throughout this specification, whenever a part is said to "include" a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise specifically stated.

[0038] The terms "about," "substantially," and the like used in this specification are used in a meaning that is at or close to the numerical value when manufacturing and material tolerances inherent in the meanings mentioned are presented, and are used to prevent unscrupulous infringers from unfairly exploiting the disclosure in which exact or absolute values ​​are mentioned to aid understanding of the present application.

[0039] The terms “step of ~” or “step of ~” as used throughout this specification do not mean “step for ~.”

[0040] Throughout this specification, the term "combination(s) thereof" included in the expressions in the Makushi format means one or more mixtures or combinations selected from the group consisting of the components described in the expressions in the Makushi format, and means including one or more selected from the group consisting of said components.

[0041] Throughout this specification, references to “A and / or B” mean “A or B, or A and B.”

[0042] As used throughout this specification, the term "alkyl" or "alkyl group" includes linear or branched alkyl groups having 1 to 8 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms and all possible isomers thereof. For example, the alkyl or alkyl group includes a methyl group (Me), an ethyl group (Et), an n-propyl group ( n Pr), iso-profiler ( i Pr), n-butyl group ( n Bu), iso-butyl group( i Bu), tert-butyl group (tert-Bu, t Bu), sec-butyl group (sec-Bu, sec Bu), n-pentyl group ( n Pe), iso-pentyl group ( iso Pe), sec-pentyl group ( sec Pe), tert-pentyl group (t Pe), neo-pentyl group ( neo Pe), 3-pentyl group, n-hexyl group, iso-hexyl group, heptyl group, 4,4-dimethylpentyl group, octyl group, 2,2,4-trimethylpentyl group, and isomers thereof, but may not be limited thereto.

[0043] As used throughout this specification, the term "alkoxy group" includes linear or branched alkoxy groups having 1 to 5 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms, and all possible isomers thereof. For example, the alkoxy group may include, but is not limited to, a methoxy group, an ethoxy group, an n-propoxy group, an n-butoxy group, an iso-butoxy group, a sec-butoxy group, a tert-butoxy group, a cyclobutoxy group, an n-pentyloxy group, an isopentyloxy group, a sec-pentyloxy group, a tert-pentyloxy group, a neopentyloxy group, a 2-methylbutoxy group, a 1,2-dimethylpropoxy group, a 1-ethylpropoxy group, a cyclopentyloxy group, and isomers thereof.

[0044] Throughout this specification, the term “halogen group” includes fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0045] Below, the implementation examples of the present invention are described in detail, but the present invention may not be limited thereto.

[0046] A first aspect of the present invention provides a compound represented by the following chemical formula 1:

[0047] [Chemical Formula 1]

[0048] ;

[0049] In the above chemical formula 1, Cbz is a benzyloxycarbonyl group, and the R 1 Inland R 4 are, each independently, hydrogen, halogen, linear or branched C 1-5 Alkyl group, and linear or branched C1-5 It is selected from an alkoxy group. In one embodiment of the present invention, in the chemical formula 1, R 1 Inland R 4 are, each independently, hydrogen, halogen, linear or branched C 1-3 Alkyl group, and linear or branched C 1-3 It may be selected from an alkoxy group, but may not be limited thereto. In one embodiment of the present invention, in the chemical formula 1, R 1 Inland R 4 may be all hydrogen, but may not be limited thereto.

[0050] In one embodiment of the present invention, the compound may exist in a solid state at room temperature, but may not be limited thereto. In one embodiment of the present invention, the compound has a terminal portion including a 1,2,3-benzotriazole group connected to the remaining compound portion by an amide bond, and the terminal portion including the 1,2,3-benzotriazole serves as an activator to activate a carboxyl group. At this time, the amide bond is a relatively stronger bond than an ester bond. Therefore, the compound can be obtained in a stable solid state and has the advantage of being easy to store and measure.

[0051] The second aspect of the present invention provides a method for preparing a compound according to the first aspect, comprising the step of reacting H-AEEA-OH(2-(2-(2-Aminoethoxy)ethoxy)acetic acid) and benzyl chloroformate.

[0052] Detailed descriptions of parts that overlap with the first aspect of the present application have been omitted, but the contents described in the first aspect of the present application may be equally applied even if the description is omitted in the second aspect of the present application.

[0053] In one embodiment of the present invention, the reaction is a reaction of H-AEEA-OH (2-(2-(2-Aminoethoxy)ethoxy)acetic acid) and benzyl chloroformate, and may be performed under basic conditions, but may not be limited thereto. The benzyl chloroformate (Cbz-Cl) acts as a protecting group that protects an amine group. Generally, benzyl N-succinimidyl carbonate (Cbz-OSu) is used instead of benzyl chloroformate as an amine protecting group, but the use of Cbz-Osu is not suitable for the manufacturing method according to the present invention. This is because when Cbz-Osu is used instead of Cbz-Cl, HOSu is generated as a by-product in the reaction. Since HOSu has the characteristic of being soluble in water and organic solvents, it is difficult to remove, which may lower the yield of the H-AEEA-AEEA-OH manufacturing process. In addition, unlike Cbz-Cl, Cbz-Osu has the disadvantage of generating additional beta-alanine-derived impurities, thereby lowering the purity of the final product.

[0054] In one embodiment of the present invention, the manufactured Cbz-AEEA-OH is 1,2,3-benzotriazole (1,2,3-benzotriazole; wherein each of the four carbon atoms of the phenyl group is independently hydrogen or a halogen group, linear or branched C 1-5 Alkyl group, and linear or branched C 1-5The compound of the above formula 1 can be obtained through a polymerization reaction with an alkoxy group (which may be substituted with a group selected from the group consisting of alkoxy). Here, a coupling agent can be used, and as a non-limiting example, at least one selected from EDCI (N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, Cas No. 25952-53-8), DIC (N,N′-Diisopropylcarbodiimide; Cas No. 693-13 0), and DCC (Cas No. 538-75-0), T3P (Propylphoshonic anhydride; Cas No. 68957-94-8) can be used.

[0055] In one embodiment of the present invention, the 1,2,3-benzotriazole may be used in an amount of about 1.05 to about 1.2 equivalents relative to Cbz-AEEA-OH, and the coupling agent may be used in an amount of about 1.2 to about 1.7 equivalents relative to Cbz-AEEA-OH.

[0056] In one embodiment of the present invention, the terminal portion containing the 1,2,3-benzotriazole group functions as an activator to activate the carboxyl group. Here, the 1,2,3-benzotriazole (HBt) used in the coupling reaction has a strong bond, so that the generation of AEEA monomer due to decomposition is very small, and therefore, purification is easy and the yield is high. In addition, since the molecule is crystalline, purification through crystallization is possible, and the solid obtained through this crystallization is stable, easy to store, and easy to handle, such as by splitting, making it suitable for mass production.

[0057] Meanwhile, existing substances that have been used to activate carboxyl groups include -OSu from hydroxysuccinimide (HOSu), -OBt from hydroxy benzotriazole (HOBt), -OBn from benzyl alcohol, or -imidazole generated when using 1,1'-carbonyl diimidazole (CDI). However, since these form relatively weak bonds (ester bonds), they are usually polymerized with H-AEEA-OH or HClH-AEEA-OH in a basic solution without additional separation or purification after the reaction. In this case, some of them may be decomposed during the reaction due to the weak bond, resulting in the remaining Cbz-AEEA-OH that does not participate in the reaction, which may increase the monomer content. This may make purification difficult and reduce the yield. Furthermore, the use of HOSu can generate beta-alanine-derived impurities, which can cause problems in purification and purity. Furthermore, while structurally similar to 1,2,3-benzotriazole (HBt), the bonds formed as a result of the polymerization reaction with Cbz-AEEA-OH are different: an ester bond (reacting with HOBt) and an amide bond (reacting with HBt), respectively. The amide bond is generally known to be stronger than the ester bond.

[0058] The third aspect of the present invention provides a method for preparing H-AEEA-AEEA-OH, comprising the steps of: (a) reacting a compound represented by the following chemical formula 1 with H-AEEA-OH (2-(2-(2-Aminoethoxy)ethoxy)acetic acid) to obtain a first reaction intermediate represented by the following chemical formula 2; and (b) removing a Cbz protecting group from the first reaction intermediate to obtain H-AEEA-AEEA-OH.

[0059] [Chemical Formula 1]

[0060] ;

[0061] [Chemical Formula 2]

[0062] ;

[0063] In the above chemical formulas 1 and 2, Cbz is a benzyloxycarbonyl group, and R 1 Inland R 4 are, each independently, hydrogen, halogen, linear or branched C 1-5 Alkyl group, and linear or branched C 1-5 It is selected from an alkoxy group. In one embodiment of the present invention, the R 1 Inland R 4 are, each independently, hydrogen, halogen, linear or branched C 1-3 Alkyl group, and linear or branched C 1-3 It may be selected from an alkoxy group, but may not be limited thereto. In one embodiment of the present invention, in the chemical formula 1, R 1 Inland R 4 may be all hydrogen, but may not be limited thereto.

[0064] Detailed descriptions of parts overlapping with the first or second aspect of the present application have been omitted, but the contents described for the first or second aspect of the present application may be equally applied even if the description is omitted in the third aspect of the present application.

[0065] In one embodiment of the present invention, step (a) may be performed under basic conditions, but may not be limited thereto. Specifically, step (a) is preferably performed by adding the NaOH aqueous solution in two, three, or five installments.

[0066] In one embodiment of the present invention, the step (a) may be performed by adding the compound according to the above chemical formula 1 in 2 or more, 3 or more, or 5 times. Accordingly, the method for producing H-AEEA-AEEA-OH according to the present invention may generate less impurities (e.g., H-AEEA-OH and H-AEEA-AEEA-AEEA-OH) due to the residual active form of the reaction solution, so that the final product may have high purity. That is, when the compound is added in portions, the amount of impurities that may occur during the reaction process can be reduced. Specifically, in the existing process, the active form of Cbz-AEEA-OH is produced, and the reaction is performed by adding a base to the reaction solution and adding H-AEEA-OH or HClH-AEEA-OH without further purifying the reaction solution due to stability issues. Here, the active form of Cbz-AEEA-OH, which remains in the reaction solution for a relatively long time, is likely to be decomposed into monomers in the basic solution. On the other hand, according to one embodiment of the present invention, by dividing Cbz AEEA-Bt into a basic solution of H-AEEA-OH and adding it in portions, the possibility of decomposition is reduced, and by also dividing the added NaOH aqueous solution, the production of Cbz AEEA-OH monomer is significantly reduced (for example, to less than 1% or 0.21%).

[0067] In one embodiment of the present invention, the first reaction intermediate may be obtained in a liquid state, but may not be limited thereto.

[0068] In one embodiment of the present invention, step (a) may further include, but is not limited to, a step of purifying the synthesized first reaction intermediate before removing the Cbz protecting group. Here, the Cbz protecting group may be an amine protecting group that protects the amine of the first reaction intermediate.

[0069] In one embodiment of the present invention, the step of purifying the first reaction intermediate may include, but is not limited to, esterifying the obtained first reaction intermediate into a second reaction intermediate represented by the following chemical formula 3 and then purifying it:

[0070] [Chemical Formula 3]

[0071] ;

[0072] In the above chemical formula 3, Cbz is a benzyloxycarbonyl group, and R is C 1-8 It may be a linear or branched alkyl group, but may not be limited thereto. Preferably, R may be a methyl group.

[0073] In one embodiment of the present invention, the step of purifying the first reaction intermediate may further include washing the second reaction intermediate with an aqueous basic solution after esterifying it. The first reaction intermediate has a disadvantage in that it is difficult to separate from the main impurities (e.g., Cbz-AEEA-OH and Cbz-AEEA-AEEA-OH) generated in step (a) due to its polarity caused by the carboxyl group (COOH) at the terminal. On the other hand, the second reaction intermediate does not contain a carboxyl group at the terminal, so it has the advantage of being less likely to be tailed and adsorbed due to interaction with silica gel, thereby reducing purification interference and loss during the purification process, and thus being able to be easily purified from the main impurities. (For example, Cbz-AEEA-OH and Cbz-AEEA-AEEA-AEEA-OH can be purified to about 0.1% or less.) In addition, the second reaction intermediate has an advantage in that the byproduct (1,2,3-benzotriazole) generated in the step (a) other than the main impurities can be easily removed simply by washing with an aqueous basic solution.

[0074] In one embodiment of the present invention, step (b) may be performed through a hydrolysis reaction and a reduction reaction, but may not be limited thereto. In one embodiment of the present invention, the reduction reaction may use a Pd / C catalyst, but may not be limited thereto.

[0075] In one embodiment of the present invention, an additional crystallization purification step may be included after step (b).

[0076] In one embodiment of the present invention, in the method for producing H-AEEA-AEEA-OH, the yield of the product H-AEEA-AEEA-OH may be about 75% or more based on the compound represented by the chemical formula 1.

[0077] Hereinafter, the present invention will be described in more detail using examples. However, the following examples are provided only to help understand the present invention, and the contents of the present invention are not limited to the following examples.

[0078] [Example]

[0079] Figure 1 is a reaction scheme showing a schematic synthesis process of H-AEEA-AEEA-OH (17-Amino-10-oxo-3,6,12,15-tetraoxa-9-azaheptadecanoic acid) according to the present invention.

[0080] 1. Preparation of compound Cbz-AEEA-Bt

[0081]

[0082] Reaction solution 1 was prepared by adding 80 g (0.490 mol) of H-AEEA-OH (2-(2-(2-Aminoethoxy)ethoxy)acetic acid) to 640 mL of purified water. The reaction solution 1 was cooled to a temperature of 0°C to 10°C. After adding a 50% NaOH aqueous solution to the cooled reaction solution 1, the pH of the reaction solution 1 was adjusted to 10.5 to 11.0. After adding 101 g (0.592 mol) of benzyl chloroformate (Cbz-Cl, where Cbz is a benzyloxycarbonyl group) to the reaction solution 1, the pH of the reaction solution 1 was adjusted to 10 to 11 to prevent the pH from lowering due to HCl generated during the reaction. At this time, the Cbz-Cl was added to introduce Cbz as an amine protecting group. When the reaction of the reaction solution 1 was completed, it was washed with methyl tert-butyl ether (MTBE), and the pH of the reaction solution 1 in which the reaction was completed was adjusted to 2.2 to 2.5 by adding an aqueous hydrochloric acid solution. Thereafter, the reaction solution 1 was extracted with dichloromethane (DCM) and dried with Na2SO4 (see Fig. 2). The dried reaction solution 1 was filtered and concentrated under reduced pressure to obtain 141 g (0.474 mol, yield: 96.7%) of Cbz-AEEA-OH. The obtained Cbz-AEEA-OH was analyzed by HPLC (High-performance liquid chromatography), and the result graph is shown in Fig. 2, and the graph information of Fig. 2 is shown in Table 1 below.

[0083] NumberRetention Time (unit: min)TypeWidth(min)Area(mAU * s)Height(mAU)Area Ratio(%)118.504Mixed0.09295569.99072999.7134499.2439224.492Mixed0.080411.757142.437830.2095324.659Mixed0.077530.680136.594070.5466

[0084]

[0085] To 9 L of DCM, 600 g (2.02 mol) of Cbz-AEEA-OH obtained from the above reaction, 289 g (2.43 mol) of 1,2,3-benzotriazole (Bt), and 479 g (6.28 mol) of pyridine were added to prepare a reaction solution 2. Thereafter, the reaction solution 2 was cooled to 5 to 10°C. 2.19 kg (3.43 mol) of a 50% T3P (Propylphosphonic Anhydride) solution was added to the cooled reaction solution 2, and the mixture was stirred at room temperature for 1 hour. When the reaction of the reaction solution 2 was completed, the reaction solution 2 was slowly added to a mixture of DCM and purified water cooled to 5 to 10°C, stirred, and the organic layer was separated. The separated organic layer was washed with aqueous NaHSO4 solution, aqueous Na2CO3 solution, and aqueous NaCl solution, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue concentrated under reduced pressure was crystallized with DCM and MTBE to obtain the compound (Cbz-AEEA-Bt) (777 g, 1.95 mol, yield: 96.5%) (see Figure 3).

[0086] The NMR values ​​of the compound Cbz-AEEA-Bt identified in Figure 3 were as follows: 1H NMR (DMSO-d6, 400 MHz) δ 8.29 (dd,J= 11.2, 8.4 Hz, 2H), 7.83 (t,J= 7.6 Hz, 1H), 7.66 (t,J= 7.4 Hz, 1H), 7.41-7.30 (m, 5H), 5.28 (s, 2H), 5.04 (s, 2H), 3.84 (t,J= 4.0 Hz, 2H), 3.67 (t,J= 4.6 Hz, 2H) 3.50 (t,J= 6 Hz, 2H), 3.19 (q,J= 6.0 Hz, 2H).

[0087] 2. Preparation of the first reaction intermediate (Cbz-AEEA-AEEA-OH)

[0088]

[0089] To H-AEEA-OH (345 g, 2.11 mol) was added 1.5 L of acetonitrile and 350 mL of DCM to prepare a reaction solution 3. After cooling the reaction solution 3 to a temperature of 5 to 10°C, a 20% NaOH aqueous solution (317 g, 1.59 mol) was added. Thereafter, 420 g (1.05 mol) of Cbz-AEEA-Bt obtained in Example 1 was added to the reaction solution 3 in three portions at 30-minute intervals, and the temperature of the reaction solution 3 was maintained and stirred for 90 minutes. To the stirred reaction solution 3, 106 g (0.530 mol) of 20% NaOH aqueous solution was added, and 280 g (0.703 mol) of Cbz-AEEA-Bt obtained in Example 1 was added in two portions at 30-minute intervals, and the temperature of the reaction solution 3 was maintained and stirred for 2 hours. When the reaction of the reaction solution 3 was completed, the pH of the reaction solution 3 was adjusted to 2.2 to 2.5 by adding an aqueous hydrochloric acid solution, and then the mixture was concentrated under reduced pressure to remove acetonitrile. The residue obtained by concentration under reduced pressure was diluted with DCM, washed three times with a 10% aqueous NaCl solution, and the organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the first reaction intermediate (Cbz-AEEA-AEEA-OH). Additionally, the content of Cbz-AEEA-OH, which did not participate in the reaction and existed in a monomeric state, was approximately 0.21% (see peak 2 in Table 2 below). The obtained Cbz-AEEA-AEEA-OH was analyzed by HPLC, and the resulting graph is shown in Figure 4. The graph information in Figure 4 is summarized and shown in Table 2 below.

[0090] NumberRetention Time (unit: min)TypeWidth(min)Area(mAU * s)Height(mAU)Area Ratio(%)117.612Mixed0.09146291.510251146.7583099.7901218.264Mixed0.076313.235262.890580.2099

[0091] 3. Preparation of the second reaction intermediate (Cbz-AEEA-AEEA-OMe)

[0092]

[0093] In Example 2, 7 L of methanol was added to the synthesized Cbz-AEEA-AEEA-OH, and the mixture was cooled to 5 to 10°C to prepare a reaction solution 4. 382 g (3.52 mol) of trimethylsilyl chloride was slowly added to the reaction solution 4, and the mixture was stirred at room temperature for 2 hours. When the reaction of the above reaction scheme 4 was completed, the reaction solution 4 was concentrated under reduced pressure, and the concentrated residue was diluted with 7 L of DCM. The diluted organic layer was washed four times with a 10% aqueous solution of Na2CO3 and dried over Na2SO4. Thereafter, the dried organic layer was filtered and concentrated under reduced pressure again. The residue concentrated under reduced pressure twice was purified using a SiO2 column to obtain 683 g (1.50 mol) of the second reaction intermediate (Cbz-AEEA-AEEA-OMe). The yield of the second reaction intermediate was 85.2%, calculated based on the total first reaction intermediate used in the reaction.

[0094] 4. Preparation of H-AEEA-AEEA-OH

[0095]

[0096] In Example 3, 175 g (0.383 mol) of the second reaction intermediate (Cbz-AEEA-AEEA-OMe) synthesized was added 175 mL of tetrahydrofuran (THF) and 350 mL of purified water, and the mixture was cooled to 5 to 15°C to prepare a reaction solution 5. To the cooled reaction solution 5, 32.3 g (0.404 mol) of a 50% NaOH aqueous solution was added in five portions. When the reaction of the reaction solution 5 was completed, the pH of the reaction solution 5 was adjusted to 12 with an aqueous hydrochloric acid solution, and then concentrated under reduced pressure to remove THF. The concentrated residue under reduced pressure was washed twice with MTBE, and the pH of the residue was adjusted to 2.2 to 2.4 with an aqueous hydrochloric acid solution. The pH-adjusted residue was extracted three times with DCM, and the extracted organic layer was dried over Na2SO4. The dried organic layer was filtered and concentrated under reduced pressure to obtain 165 g (0.373 mol, 97.4%) of Cbz-AEEA-AEEA-OH.

[0097]

[0098] To 165 g (0.373 mol) of Cbz-AEEA-AEEA-OH obtained from the above reaction, 1 L of methanol, 0.5 L of purified water, and 3.3 g of Pd / C were added, and the mixture was stirred under 0.4 MPa of hydrogen for more than 6 hours to prepare a reaction solution 6. Upon completion of the reaction of reaction solution 6, methanol was first removed by concentration under reduced pressure, and then Pd / C was removed by filtering with diatomaceous earth (celite). The filtrate from which Pd / C was removed was concentrated under reduced pressure, and then purified by recrystallization to obtain 107 g (0.347 mol, yield: 93.0% based on 165 g of the Cbz-AEEA-AEEA-OH) of H-AEEA-AEEA-OH (see Fig. 5).

[0099] The obtained H-AEEA-AEEA-OH was analyzed by HPLC, and the result graph is shown in Fig. 5. In addition, the graph information of Fig. 5 is summarized in Table 3 below. According to Fig. 5 and Table 3, no beta-alanine-derived impurities (e.g., Beta-Ala-AEEA-OH, Beta-Ala-AEEA-AEEA-OH) were detected in the obtained H-AEEA-AEEA-OH. In addition, it was confirmed that the obtained H-AEEA-AEEA-OH contained 0.008% and 0.013% of the impurities H-AEEA-OH and H-AEEA-AEEA-AEEA-OH, respectively, indicating that H-AEEA-AEEA-OH was synthesized with high purity through the process of the above example.

[0100] NumberPeak NameRetention Time (unit: min)Area (pA * s)Height (pA)Area Ratio (%)Resolution (USP)12.3180.0050.0780.02019.3324.3950.0010.0100.00321.6436.7970.0020.0220.0074.4847.2280.0010.0160.0035.685H-AEEA-OH7.8250.0020.0340.0080.7667.9150.0070.0990.02615.3679.6220.0030.0 410.0118.61Not detectedBeta-Ala-AEEA-----8H-AEEA-AEEA-OH11.62325.302133.77599.7772.34912.1750.0200.2430.0771 7.86Not detectedBeta-Ala-AEEA-AEEA-----10H-AEEA-AEEA-AEEA-OH14.1530.0030.0520.0131.831114.3550.0140.1780.053-

[0101] [Comparative example]

[0102] In order to confirm the effectiveness of the H-AEEA-AEEA-OH manufacturing method according to the example and FIG. 1, N-(Benzyloxycarbonyloxy)succinimide (Cbz-OSu) was used instead of benzyl chloroformate (Cbz-Cl) of Example 1.

[0103] 1. Preparation of Cbz-AEEA-OH using Cbz-Osu

[0104]

[0105] Solution 1 was prepared by adding 50 mL of THF, 10 mL of purified water, and 6.50 g (61.3 mmol) of Na2CO3 to H-AEEA-OH (10 g, 61.3 mmol), and then cooled to 15°C. 16.8 g (67.4 mmol) of Cbz-OSu was added to the solution, and the mixture was stirred for 6 hours while maintaining the temperature at 15°C. Upon completion of the reaction of Solution 1, the pH of Solution 1 was adjusted to 2.2 to 2.5 with aqueous hydrochloric acid, and then Solution 1 was extracted with DCM (50 mL × 3). The extracted organic layer was washed with water (20 mL × 2) and dried over Na2SO4. The dried organic layer was then filtered and concentrated under reduced pressure to obtain 20.3 g (68.3 mmol, 111%) of Cbz-AEEA-OH. The composition above was analyzed by HPLC (High-performance liquid chromatography) and is shown in Fig. 6. In addition, the graph information of Fig. 6 is summarized in Table 4 below. As a result of comparing with the HLPC analysis result of Cbz-AEEA-OH prepared according to Example 1 (Fig. 2 and Table 1), additional peaks indicating impurities were confirmed in Cbz-AEEA-OH prepared according to the comparative example (Fig. 6). In addition, while the area of ​​the peak indicating Cbz-AEEA-OH in Fig. 2 was about 99.2%, the area of ​​the peak indicating Cbz-AEEA-OH in Fig. 6 was confirmed to be about 77.6%, indicating that the purity of Cbz-AEEA-OH of the comparative example is lower than that of Example 1.

[0106] Number Retention Time (unit: min) Type Width (min) Area (mAU *) s)Height (mAU)Area ratio (%)13.000Mixed0.0586562.84692159.955227.6457214.783Mixed0.1123105.7351615.689311.4363317.312Mixed0.136342.207675.162340.5734417.799Mixed0.098237.896146.431360.5148518.031Mixed0.085712.834452.495860.1743618.376Mixed0.09095712.503421046.957 4077.5988718.709Mixed0.106666.9248610.460620.9091820.049Mixed0.088215.493752.928550.2105920.508Mixed0.123912.344131.660140.16771021.448Mixed0.08245.529421.117840.07511122.594Mixed0.0905751.40265138.4092110.20711227.306Mixed0.114835.865195.208020.4872

[0107] 2. Preparation of Cbz-AEEA-AEEA-OH using Cbz-AEEA-OH

[0108]

[0109] Solution 2 was prepared by adding 6.96 g (60.5 mmol) of HOSu (Hydroxysuccinimide) and 85 mL of DCM to 19.0 g (crude, 57.6 mmol) of Cbz-AEEA-OH obtained in Comparative Example 1. Solution 2 was cooled to 5°C, EDCI (13.3 g, 69.4 mmol) was added, and solution 2 was stirred at 5°C for 2 hours. When the reaction was complete, 7.44 g (57.6 mmol) of DIPEA (N,N-Diisopropylethylamine) and 14.1 g (86.4 mmol) of H-AEEA-OH were added to solution 2, and the mixture was stirred again at 5°C for 3 hours. When the reaction was complete, the pH of solution 2 was adjusted to 2.2 to 2.5 with aqueous hydrochloric acid, and extracted with 80 mL of DCM. The extracted organic layer was washed with water (35 mL × 2) and dried over Na2SO4. The dried organic layer was filtered and concentrated under reduced pressure to obtain 25.0 g (56.5 mmol, 98.1%) of Cbz-AEEA-AEEA-OH. The obtained Cbz-AEEA-AEEA-OH was analyzed by HPLC as shown in FIG. 7, and the graph information of FIG. 7 is shown in Table 5 below. As a result of comparing FIG. 7 with the HLPC analysis result (FIG. 4 and Table 2) of Cbz-AEEA-AEEA-OH prepared according to Example 2, the area of ​​the peak indicating Cbz-AEEA-AEEA-OH in FIG. 4 was confirmed to be about 99.8%, whereas the area of ​​the peak indicating Cbz-AEEA-AEEA-OH in FIG. 7 was confirmed to be about 64.0%. In addition, in Fig. 7, the content of Cbz-AEEA-OH is confirmed to be about 25.0%, indicating that the purity of Cbz-AEEA-AEEA-OH of the comparative example is very low (see No. 6 in Table 5).

[0110] NumberRetention Time (unit: min)TypeWidth(min)Area(mAU * s)Height(mAU)Area Ratio(%)13.001Mixed0.054576.8866023.515180.9829214.789Mixed0.094541.391187.302600.5291317.273Mixed0.091050.295199.212390.643041 7.472 Mixed 0.0778405.2770186.852835.1811517.803 Mixed 0.08675004.76611962.3137236.9813618.385 Mixed 0.08391962.75378389.8171425.0920718.717 Mixed 0.089737.581926.985860.4805818.840Mixed 0.072618.990674.362100.2428919.752Mixed 0.085099.3084319.482521.26961020.085Mixed 0.090835.8049 16.569830.45771120.527Mixed0.097229.362975.034170.37541221.177Mixed0.084843.248388.501680.55291321.460Mixed0.082116.563723.364510.2118

[0111] The above description of the present invention is for illustrative purposes only, and those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single entity may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined manner.

[0112] The scope of the present invention is indicated by the claims described below rather than the detailed description above, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. A compound represented by the following chemical formula 1: [Chemical Formula 1] ; In the above chemical formula 1, Cbz is a benzyloxycarbonyl group, The above R 1 Inland R 4 are, each independently, hydrogen, halogen, linear or branched C 1-5 Alkyl group, and linear or branched C 1-5 It is selected from alkoxy groups.

2. In paragraph 1, In the above chemical formula 1, the R 1 Inland R 4 A compound in which all of them are hydrogen.

3. In paragraph 1, A compound that exists in a solid state at room temperature.

4. Step of reacting H-AEEA-OH (2-(2-(2-Aminoethoxy)ethoxy)acetic acid) and benzyl chloroformate A method for producing a compound according to claim 1, comprising:

5. In paragraph 4, A method for producing a compound, wherein the above reaction is performed under basic conditions. 6.(a) a step of reacting a compound represented by the following chemical formula 1 with H-AEEA-OH(2-(2-(2-Aminoethoxy)ethoxy)acetic acid) to obtain a first reaction intermediate represented by the following chemical formula 2; and (b) a step of removing the Cbz protecting group from the first reaction intermediate to obtain H-AEEA-AEEA-OH; A method for producing H-AEEA-AEEA-OH, comprising: [Chemical Formula 1] ; [Chemical Formula 2] ; In the above chemical formulas 1 and 2, Cbz is a benzyloxycarbonyl group, The above R 1 Inland R 4 are, each independently, hydrogen, halogen, linear or branched C 1-5 Alkyl group, and linear or branched C 1-5 It is selected from alkoxy groups.

7. In paragraph 6, In the above chemical formula 1, the R 1 Inland R 4 A manufacturing method in which all of them are hydrogen.

8. In paragraph 6, A manufacturing method wherein the above step (a) is performed under basic conditions.

9. In paragraph 6, A manufacturing method wherein the compound represented by the above chemical formula 1 is divided into two or more times and mixed with H-AEEA-OH to react.

10. In paragraph 6, A manufacturing method wherein the first reaction intermediate is obtained in a liquid state.

11. In paragraph 6, A manufacturing method, wherein the step (a) further comprises a step of purifying the obtained first reaction intermediate.

12. In paragraph 11, The step of purifying the first reaction intermediate is: A manufacturing method comprising esterifying the obtained first reaction intermediate into a second reaction intermediate represented by the following chemical formula 3 and then purifying it: [Chemical Formula 3] ; In the above chemical formula 3, Cbz is a benzyloxycarbonyl group, and R is C 1-8 It is a linear or branched alkyl group.

13. In paragraph 12, A manufacturing method wherein the above R is a methyl group.

14. In paragraph 6, A manufacturing method wherein the above step (b) is performed through a hydrolysis reaction and a reduction reaction.

15. In paragraph 14, A manufacturing method wherein the above reduction reaction uses a Pd / C catalyst.

16. In paragraph 6, A manufacturing method wherein the yield of H-AEEA-AEEA-OH, which is a product of the manufacturing method, is 75% or more based on the compound represented by the above chemical formula 1.

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