New acid addition salts and crystalline forms of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol

Abstract

The present invention relates to the novel (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride, (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol oxalate ... The present invention provides crystalline forms of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol maleate, (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol palmitate and (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride. These novel acid addition salts and crystalline forms have low hygroscopicity and excellent stability under high temperature and high humidity conditions, and can be used pharma- ceuticals.

Description

[Technical field]

[0001] The present invention relates to pharma- ceutically acceptable acid addition salts and crystalline forms of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol. [Background technology]

[0002] The selection of pharma- ceutically acceptable salts and their polymorphs is a critical step in the research and development process of new drugs, because a particular salt form and its polymorphs can have a significant impact on the development of the bulk drug product manufacturing process, the finished drug product design and dosage form.

[0003] Specifically, the type of salt and its polymorphism can affect the recrystallization yield, process speed, and purity in the final stage of raw drug production, i.e., the recrystallization and purification process. In particular, the speed of the crystallization process can vary depending on the size and shape of the crystals, so the selection of the salt and polymorphism is important in terms of the productivity and production costs of the raw drug.

[0004] In addition, the addition of salts to certain drugs can be a way to change the physicochemical and biological properties of the drug without changing its chemical structure. In pharmaceutical terms, physicochemical properties such as hygroscopicity, stability, solubility, particle flowability, and dissolution rate are affected by the salt form and its polymorphic form. Therefore, the salt form and its polymorphic form are factors that determine the production process, production and storage conditions, and expiration date of the finished drug product.

[0005] Therefore, there is a need for continued research into the selection of salts and crystalline polymorphs that can exhibit better stability and better bioavailability in producing formulations of specific drugs.

[0006] Meanwhile, the present inventors have identified (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol as a novel active pharmaceutical substance that can inhibit prolyl-tRNA synthetase (PRS) enzyme, which is a member of the aminoacyl-tRNA synthetase (ARS) family of enzymes and plays a role in activating amino acids for protein synthesis.

[0007] Such PRS exists in the form of EPRS (Glutamyl-Prolyl-tRNA Synthetase) in the form of a multisynthetase complex (MSC). Among various MSCs, EPRS functions as a translational silencer that suppresses the production of VEGF A (vascular endothelial growth factor A), a core factor in angiogenesis, and has been reported to be closely related to various solid cancers (Nat. Rev. Cancer, 2011, 11, 708-718). The (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol active pharmaceutical substance is useful as a PRS (prolyl-tRNA synthetase) anti-fibrotic agent, anti-inflammatory agent, autoimmune therapeutic agent, or next-generation anti-cancer agent that can be used alone or in combination with existing targeted anti-cancer agents.

[0008] Therefore, the present inventors have conducted research into the selection of a pharma- ceutically acceptable salt and its crystalline polymorphism that can improve the bioavailability and stability of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol. As a result, it has been found that when a specific acid addition salt form is prepared, the hygroscopicity can be reduced while the stability under harsh conditions can be improved, and thus the present invention has been completed. Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention provides a pharma- ceutically acceptable acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol which has low hygroscopicity and excellent stability.

[0010] The present invention also provides a crystalline form of a pharma- ceutically acceptable acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, which has low hygroscopicity and excellent stability. [Means for solving the problem]

[0011] In order to solve the above problems, the present invention provides Provided is the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

[0012] The present invention also provides a method for producing a method for manufacturing a semiconductor device comprising the steps of: Provided is an acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, wherein the acid addition salt is an oxalate, maleate, or palmitate salt.

[0013] The present invention also provides a method for producing a method for manufacturing a semiconductor device comprising the steps of: Provided is a crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

[0014] Meanwhile, the novel active pharmaceutical substance (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol is represented by the following chemical formula 1: <Chemical formula 1> JPEG0007644873000001.jpg2464 The (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol and its pharma- ceutically acceptable salts can inhibit PRS enzyme activity while exhibiting reduced toxicity, and can therefore be used to prevent or treat diseases caused by abnormal PRS activity. Thus, the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol and its pharma- ceutically acceptable salts can be usefully applied to the prevention or treatment of cancer, inflammatory diseases, autoimmune diseases, or fibrosis caused by abnormal PRS activity.

[0015] Here, the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol and its pharma- ceutically acceptable salts are acid addition salts of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol. Such acid addition salts can be prepared by reacting the free base of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol and the acid to be added in a specific ratio.

[0016] In addition, the acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol may exhibit a crystalline form. The crystalline form of the acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol may be prepared by various crystallization methods, such as evaporation crystallization, solution crystallization, reaction crystallization, solvent-mediated polymorphic transformation, and solid-state polymorphic transformation, selected according to the thermodynamic and kinetic properties of the salt.

[0017] In this case, the crystal form of the acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol can be confirmed by X-ray powder diffraction analysis. Specifically, the crystal form is classified by the diffraction angle (2θ) showing a characteristic peak in the X-ray powder diffraction pattern and the intensity of the peak corresponding to each diffraction angle (2θ). Here, the diffraction angle (2θ) can vary within ±0.2°, or preferably ±0.1°, depending on various factors such as the manufacturing technique of the measurement sample, the fixing procedure of the measurement sample, and the measurement device.

[0018] The acid addition salts of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol and their crystal forms are also classified by the endothermic onset temperature and the endothermic temperature showing the maximum endothermic peak in differential scanning calorimetry. Here, the temperature can vary within ±3°C, preferably ±2°C, or more preferably ±1°C depending on various factors such as the manufacturing technique of the measurement sample, the measurement equipment, and the rate of temperature change.

[0019] The present invention will be described in detail below.

[0020] (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride, oxalate, maleate and palmitate salts On the other hand, it is known that the free base form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol is not stable. Therefore, in order to improve the stability of such a compound, the dihydrochloride of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol as an acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol has been studied.

[0021] However, (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride is highly hygroscopic and therefore unsuitable for use in pharmaceutical preparations. As a result of extensive research by the present inventors into acid addition salts that have low hygroscopicity but can improve the stability of a compound, the inventors were able to confirm that when the compound is combined with a specific type of acid at a specific equivalent ratio, the hygroscopicity can be reduced while the stability under harsh conditions can be improved.

[0022] Specifically, a preferred acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol is the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

[0023] The monohydrochloride of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol has fewer ionizable hydrogen ions and chloride ions than the dihydrochloride, and thus reduces binding with moisture in the air. Therefore, it can exhibit lower hygroscopicity and superior stability under harsh conditions compared to (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride.

[0024] The (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride salt can exhibit an endothermic onset temperature of 279.68±3°C and a maximum endothermic peak at an endothermic temperature of 281.04±3°C in differential scanning calorimeter analysis. In contrast, the dihydrochloride of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol shows two endothermic peaks in differential scanning calorimetry (a first peak with an endothermic onset temperature of 233.09°C and an endothermic temperature of the maximum endothermic peak of the peak of 242.09°C, and a second peak with an endothermic onset temperature of 271.95°C and an endothermic peak with an endothermic temperature of 275.07°C), which can be confirmed from Test Example 2 described later. Therefore, it can be seen that the monohydrochloride of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, which shows a higher endothermic onset temperature, is more stable to heat than the dihydrochloride.

[0025] The (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride may be in a crystalline form. The monohydrochloride being in a crystalline form is advantageous in terms of improved stability and improved hygroscopicity compared to an amorphous form or a mixture of a crystalline form and an amorphous form.

[0026] Further preferred acid addition salts of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol are as follows: (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol oxalate; (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol maleate; and (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol palmitate.

[0027] In this case, in terms of improving hygroscopicity, the acid addition salt is more preferably a dioxalate, a hemi-oxalate, a hemi-maleate, or a monopalmitate.

[0028] In one embodiment, the acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol is a dioxalate salt. The dioxalate salt is produced by a production method to produce dioxalates I and II.

[0029] Specifically, the dioxalate salt, in other words, dioxalate salt I, can exhibit an endothermic onset temperature of 210.42±3° C. and a maximum endothermic peak at an endothermic temperature of 213.66±3° C. in differential scanning calorimetry.

[0030] Moreover, the dioxalate, in other words, the dioxalate II, can exhibit an endothermic onset temperature of 193.84±3° C. and a maximum endothermic peak at an endothermic temperature of 203.92±3° C. in differential scanning calorimetry.

[0031] In another embodiment, the acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol is the hemi-oxalate salt.

[0032] The hemi-oxalate salt can exhibit an endothermic onset temperature of 253.72±3° C. and a maximum endothermic peak at an endothermic temperature of 255.62±3° C. in differential scanning calorimetry.

[0033] In yet other embodiments, the acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol is the hemi-maleate salt.

[0034] The hemi-maleate salt can exhibit an endothermic onset temperature of 90.69±3° C. and a maximum endothermic peak at an endothermic temperature of 110.16±3° C. in differential scanning calorimetry.

[0035] In yet another embodiment, the palmitate salt can exhibit an endothermic onset temperature of 92.32±3° C. and a maximum endothermic peak at an endothermic temperature of 98.59±3° C. in differential scanning calorimetry.

[0036] On the other hand, the acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol can be prepared by a method including the following steps: 1) mixing the dihydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol with a base to produce the free base of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol; and 2) mixing and stirring the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol free base and acid in the presence of an organic solvent.

[0037] Step 1) is a step of mixing the dihydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, the preparation of which is known, with a base to prepare a free form from which hydrochloric acid has been removed, and the prepared free form is in a viscous liquid state.

[0038] In this case, the base that can be used is at least one strong base selected from the group consisting of sodium hydroxide, lithium hydroxide, and potassium hydroxide.

[0039] In addition, the pH in step 1) is preferably about 9 to about 12 in order to remove hydrochloric acid.

[0040] Furthermore, step 2) is a step of mixing and stirring the free base of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol and an acid in the presence of an organic solvent to prepare a salt in which the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol and the acid are chemically bonded.

[0041] In step 2), the acid can be used in an amount adjusted according to the bond ratio to be finally produced relative to 1 equivalent of the free base of the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol. For example, when preparing a monovalent acid addition salt in which the free base is bonded with a monovalent acid, the acid can be used in an amount of 0.8 to 1.2 equivalents, preferably 1 equivalent, relative to 1 equivalent of the free base. In addition, when preparing a divalent acid addition salt in which the free base is bonded with a divalent acid, the acid can be used in an amount of 1.8 to 2.2 equivalents, preferably 2 equivalents, relative to 1 equivalent of the free base. In addition, when preparing a hemi-acid addition salt in which a divalent free base is bonded with a monovalent acid, the acid can be used in an amount of 0.4 to 0.6 equivalents, preferably 0.5 equivalents, relative to 1 equivalent of the free base.

[0042] Here, the organic solvent may be one or more selected from the group consisting of methanol, ethanol, acetonitrile, isopropanol, butanol, tetrahydrofuran, acetone, dimethylformamide, and methyl sulfoxide. Such an organic solvent may be used in an amount (mL / g) of 5 to 60 times the volume of the free base of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol. Preferably, the organic solvent may be used in an amount (mL / g) of 20 to 50 times the volume of the free base of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

[0043] Next, the step of stirring the mixed solution of the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol free base, acid and organic solvent is carried out at a temperature of 20°C to 30°C for 30 minutes to 30 hours. At this time, the stirring speed may be within a range of 50 to 300 rpm. Within this range, a salt can be produced in high yield and purity.

[0044] The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol prepared by the above preparation method is recovered from the solution by a vacuum filtration process. If necessary, the recovered acid addition salt can be washed and vacuum dried to obtain a high purity acid addition salt. In addition, the reaction conditions such as the ratio of solvents, temperature range, process time, etc. described in the above preparation method can be adjusted depending on the solvent selected.

[0045] Alternatively, the acid addition salts of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, excluding the hydrochloride salt, can also be prepared by a process comprising the following steps: 1) mixing the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol with a base to produce the free base of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol; and 2) mixing and stirring the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol free base and an acid other than hydrochloric acid in the presence of an organic solvent.

[0046] Crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol Meanwhile, the crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol may have peaks at diffraction angles (2θ±0.2°) of 15.0°, 17.2°, 19.8°, 23.6° and 27.8° in an X-ray powder diffraction pattern.

[0047] Specifically, the crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol can have peaks at diffraction angles (2θ±0.2°) of 15.0°, 17.2°, 19.8°, 20.9°, 23.6°, 25.1°, 27.8°, 28.9° and 31.3° in an X-ray powder diffraction pattern.

[0048] More specifically, the crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol can have peaks at diffraction angles (2θ±0.2°) of 7.5°, 12.5°, 15.0°, 16.5°, 17.2°, 18.9°, 19.8°, 20.9°, 22.3°, 23.6°, 24.8°, 25.1°, 26.1°, 27.8°, 28.9°, 30.1°, 30.9°, 31.3° and 32.4° in an X-ray powder diffraction pattern.

[0049] In addition, the crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol can exhibit an endothermic onset temperature of 279.68±3°C and a maximum endothermic peak at an endothermic temperature of 281.04±3°C in differential scanning calorimetry.

[0050] The crystalline form of the monohydrochloride of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol has a different crystalline structure from the crystalline form of the dihydrochloride of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, and has fewer ionizable hydrogen ions and chloride ions than the dihydrochloride, which reduces the binding with moisture present in the air. Therefore, the crystalline form of the monohydrochloride has a higher endothermic onset temperature than the dihydrochloride, and can exhibit high thermal stability up to relatively high temperatures.

[0051] Such a crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol can be prepared using evaporative crystallization or solution crystallization.

[0052] In one embodiment, the crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol is prepared by an evaporative crystallization method comprising the steps of: 1) (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride was reacted with C 1-8 dissolving or suspending in one or more solvents selected from the group consisting of aliphatic alcohols, pentane, hexane, heptane, cyclohexane, benzene, toluene, methyl acetate, ethyl acetate, acetonitrile, acetone, tetrahydrofuran, dichloromethane, ethyl ether, ethylene glycol, propylene glycol, and butylene glycol to produce a solution or suspension; and 2) evaporating the solvent from the solution or suspension to crystallize the monohydrochloride salt.

[0053] The step 1) is a step of dissolving the monohydrochloride using a good solvent or a suspension solvent capable of completely dissolving the monohydrochloride, and is carried out at 20° C. to 30° C., preferably at room temperature. The solvent can be used in an amount (mL / g) that is 5 to 200 times the weight of the monohydrochloride.

[0054] At this time, the above C 1-8 The aliphatic alcohol may be methanol, ethanol, propanol, isopropanol, n-butanol, or n-octanol.

[0055] Step 2) is a step of crystallizing the monohydrochloride salt by evaporating the solvent from the solution or suspension prepared in step 1) to make the solution or suspension supersaturated, and is carried out at a temperature of 20°C to 30°C, preferably at room temperature for 1 to 4 days.

[0056] Alternatively, the crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol can be prepared by a solution crystallization method comprising the steps of: 1) (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride was reacted with C 1-8 dissolving in an aliphatic alcohol solvent to produce a solution; and 2) Adding C to the solution 1-8 and crystallizing the hydrochloride salt by adding and stirring one or more crystallization solvents selected from the group consisting of aliphatic alcohols, pentane, hexane, heptane, cyclohexane, benzene, toluene, methyl acetate, ethyl acetate, acetonitrile, acetone, tetrahydrofuran, dichloromethane, ethyl ether, ethylene glycol, propylene glycol, and butylene glycol.

[0057] The step 1) is a step of dissolving the monohydrochloride using a good solvent capable of completely dissolving the monohydrochloride, which is carried out at room temperature, and the solvent can be used in an amount (mL / g) of 5 to 200 times the weight of the monohydrochloride.

[0058] The step 2) is a step of crystallizing the hydrochloride by adding an anti-solvent to the solution prepared in the step 1) to change the solubility, and the step is carried out at a temperature of 20°C to 30°C for 1 to 4 days.

[0059] The crystallization solvent can be used in an amount (mL / g) that is 5 to 200 times, or preferably 5 to 150 times, the volume of the monohydrochloride salt, and the volume ratio of the solvent in step 1) to the crystallization solvent in step 2) can be 1:1 to 1:2. Within this range, high-yield, high-purity crystals can be produced without economic losses due to increased crystallization time and excessive use of solvent.

[0060] The crystals produced by the evaporation crystallization method or the solution crystallization method are recovered from the solution by a vacuum filtration process. If necessary, the recovered crystals can be washed and vacuum dried to obtain a crystalline form of hydrochloride with high purity. In addition, the reaction conditions such as the ratio of the solvent, the temperature range, and the process time described in the preparation method can be adjusted depending on the solvent selected.

[0061] On the other hand, the present invention relates to the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, the oxalate salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, and the oxalate salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol. The present invention provides a pharmaceutical composition comprising one or more crystalline forms selected from the group consisting of maleate salt of piperidin-3-ol, palmitate salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol and monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

[0062] More specifically, the present invention relates to the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, the oxalate salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, the monomer salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, The present invention provides a pharmaceutical composition for preventing or treating cancer, an inflammatory disease, an autoimmune disease, or a fibrotic disease, comprising one or more crystalline forms selected from the group consisting of a crystalline salt of 2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, a palmitate salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, and a monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

[0063] Such pharmaceutical compositions may contain commonly used pharma- ceutically acceptable carriers. The carriers are those commonly used in formulations, including, but not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above ingredients, the pharmaceutical compositions may additionally contain lubricants, wetting agents, sweeteners, flavorings, emulsifiers, suspending agents, preservatives, and the like.

[0064] The pharmaceutical compositions may be administered orally or parenterally, including intravenous, intramuscular, intraperitoneal, subcutaneous and transdermal administration.

[0065] In this case, the pharmaceutical composition can be administered in a therapeutically effective amount, for example, about 0.001 mg / kg to about 100 mg / kg per day. The dosage can be changed depending on the formulation method, administration method, age, weight, sex, condition, food, administration time, administration route, excretion rate, or reaction sensitivity of the patient.

[0066] The pharmaceutical composition may be prepared in a unit dose form or in a multi-dose container by formulating it with a pharma- ceutically acceptable carrier and / or excipient in a manner that can be easily carried out by a person skilled in the art to which the present invention pertains. In this case, the dosage form may be any suitable form for pharmaceutical preparation, including oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations such as ointments and creams, suppositories, and sterile injection solutions, and may further include a dispersant or stabilizer. Effect of the Invention

[0067] The novel acid addition salts and crystalline forms of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to the present invention have low hygroscopicity and excellent stability under high temperature and humidity conditions, making them suitable for pharmaceutical use. [Brief description of the drawings]

[0068] [Figure 1] FIG. 1 shows the results of differential scanning calorimetry and thermogravimetry of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1. [Diagram 2] FIG. 1 shows the differential scanning calorimetry and thermogravimetry results of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dioxalate I prepared in Example 1-2. [Diagram 3]FIG. 1 shows the differential scanning calorimetry and thermogravimetry results of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dioxalate II prepared in Example 1-3. [Figure 4] FIG. 1 shows the differential scanning calorimetry and thermogravimetric analysis results of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol hemi-oxalate prepared in Example 1-4. [Diagram 5] FIG. 1 shows the differential scanning calorimetry and thermogravimetry results of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol hemi-maleate prepared in Example 1-5. [Figure 6] FIG. 1 shows the results of differential scanning calorimetry and thermogravimetry of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol palmitate prepared in Example 1-6. [Figure 7] FIG. 2 shows the results of differential scanning calorimetry and thermogravimetry of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 1. [Figure 8] FIG. 1 shows the results of differential scanning calorimetry after one week of stress testing against temperature for (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1. [Figure 9] FIG. 1 shows the results of differential scanning calorimetry of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 after 8 weeks of stress testing against temperature. [Figure 10]FIG. 1 shows the results of differential scanning calorimetry of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 1 after one week of stress testing against temperature. [Figure 11] FIG. 1 shows the results of differential scanning calorimetry of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 1 after 8 weeks of stress testing against temperature. [Figure 12] FIG. 2 shows the X-ray powder diffraction pattern for the crystalline form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 2-1. [Figure 13] FIG. 1 shows the X-ray powder diffraction pattern for the crystalline form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 2. [Figure 14] FIG. 2 shows the results of differential scanning calorimetry and thermogravimetry for the crystalline form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 2-1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0069] Hereinafter, preferred examples will be presented for understanding the present invention. However, the following examples are merely provided for easier understanding of the present invention, and the detailed description of the present invention is not limited to the following examples.

[0070] Preparation Example: Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) JPEG0007644873000002.jpg53154

[0071] Step 1-1) Preparation of compound 1-3 (tert-butyl (2R,3S)-3-((tert-butyldimethylsilyl)oxy)-2-(3-((3,4-dichloro-2-nitrophenyl)amino)propyl)piperidine-1-carboxylate) Compound 1-1, tert-butyl (2R,3S)-2-(3-aminopropyl)-3-((tert-butyldimethylsilyl)oxy)piperidine-1-carboxylate (1.0 g, 2.7 mmol) was dissolved in N,N-dimethylformamide (25 mL, 0.67 M), and then compound 1-2, 1,2-dichloro-4-fluoro-3-nitrobenzene (629.97 mg, 3.0 mmol) and N,N-diisopropylethylamine (0.93 mL, 5.4 mmol) were added and heated and stirred at 60 ° C for 2 hours. When the reaction was completed, it was diluted with ethyl acetate and washed with saturated sodium chloride solution. The organic layer was collected, dried over sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (hexane: ethyl acetate = 5: 1) to obtain compound 1-3 (1.2 g, 81% yield).

[0072] 1 H NMR (500 MHz, MeOD): δ 7.43 (d, 1H), 6.89 (d, 1H), 4.08 (s, 1H), 3.94 (s, 1H), 3.76 (s, 1H), 3.24 (m, 2H), 2.76 (s, 1H), 1.86 (m, 1H), 1.74 (m, 2H), 1.58 (m, 3H), 1.48 (s, 10H), 1.45 (s, 1H), 0.90 (s, 9H), 0.07 (d, 6H)

[0073] Step 1-2) Preparation of compound 1-4 (tert-butyl (2R,3S)-2-(3-((2-amino-3,4-dichlorophenyl)amino)propyl)-3-((tert-butyldimethylsilyl)oxy)piperidine-1-carboxylate) Compound 1-3 (2.6 mg, 4.9 mmol) obtained in step 1-1 was dissolved in methanol (25 mL, 0.2 M), and then an appropriate amount of Raney nickel was added, and a hydrogen balloon was connected, followed by stirring at room temperature for 1 hour. Upon completion of the reaction, the reaction solution was filtered through Celite and concentrated under reduced pressure to obtain compound 1-4. The obtained compound was used in the next step without purification.

[0074] Step 1-3) Preparation of compound 1-5 (tert-butyl (2R,3S)-3-((tert-butyldimethylsilyl)oxy)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidine-1-carboxylate) Compound 1-4 obtained in step 1-2 was dissolved in toluene (30 mL, 0.16 M), and then trimethyl orthoformate (1.6 mL, 14.6 mmol) and paratoluenesulfonic acid (168 mg, 0.98 mmol) were added and heated and stirred at 50° C. for 6 hours. When the reaction was completed, the solvent was removed, diluted with ethyl acetate, and washed with saturated sodium chloride solution. The organic layer was collected, dried over magnesium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (hexane:ethyl acetate=1:1) to obtain compound 1-5 (1.8 g, yield 74%).

[0075] 1 H NMR (500 MHz, MeOD): δ 8.30 (s, 1H), 7.56 (d, 1H), 7.43 (d, 1H), 4.30 (m, 2H), 4.17 (s, 1H), 4.05 (s, 1H), 3.91 (d, 1H), 3.73 (s, 1H), 2.68 (s, 1H), 1.87 (s, 3H), 1.70 (t, 2H), 1.55 (d, 1H), 1.45 (m, 10H), 1.42 (s, 1H), 0.90 (s, 9H), 0.07 (d, 6H)

[0076] Step 1-4) Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride Compound 1-5 (1.8 g, 3.6 mmol) obtained in step 1-3 was dissolved in a small amount of tetrahydrofuran, and then 4N hydrogen chloride dioxane solution (30 mL, 0.12 M) was added and stirred at room temperature for 12 hours. When the reaction was completed, the reaction solution was concentrated under reduced pressure to remove the solvent, and a small amount of methanol was added to dissolve the compound, followed by crystallization with diethyl ether to obtain compound 1-6, (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol 2HCl (26 mg, yield 81%).

[0077] 1 H NMR (500 MHz, MeOD): δ 9.67 (s, 1H). 8.02 (d, 1H), 7.82 (d, 1H), 4.62 (m, 2H), 3.60 (m, 1H), 3.28 (m, 1H), 2.99 (m, 2H), 2.25 (m, 2H), 2.08 (m, 2H), 1.99 (m, 1H), 1.78 (m, 2H), 1.54 (m, 1H)

[0078] Step 1-5) Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) 200 mg of compound 1-6 obtained in step 1-4 was placed in a beaker and dissolved in 2 mL of water. 1 mL of 1N NaOH was added to the beaker and mixed well, then the mixture was placed in a separatory funnel, at which point the pH was about 9.5. The mixture was extracted with methylene chloride (MC) and concentrated by rotary evaporation, after which the remaining solvent was removed under vacuum to obtain a viscous liquid (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (150 mg, 92% yield).

[0079] Molecular weight 328.24 1H-NMR (500 MHz, DMSO): δ 8.39 (s, 1H). 7.65 (d, 1H), 7.46 (d, 1H), 4.47 (d, 1H), 4.25 (t, 2H), 2.85 (m, 1H), 2.76 (d, 1H), 2.28 (m, 1H), 2.07 (m, 1H), 1.95 (m, 1H), 1.82 (m, 1H), 1.73 (m, 2H), 1.50 (m, 1H), 1.27 (m, 1H), 1.12 (m, 2H)

[0080] In the following examples, (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) prepared in the above Preparation Example was used.

[0081] Example 1-1: Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride JPEG0007644873000003.jpg2479 4 mL of ethanol was added to 100 mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) prepared in the above Preparation Example, and 32 mg (1 eq.) of 35% HCl solution was added and stirred at room temperature for 10 minutes. The resulting white solid was filtered and washed with ethyl acetate to obtain the title compound (103 mg, yield: 93%).

[0082] Molecular weight 364.70 1H-NMR (500 MHz, DMSO): δ 8.45 (s, 1H). 7.71 (d, 1H), 7.47 (d, 1H), 5.40 (d, 1H), 4.32 (m, 2H), 3.41 (m, 1H), 3.08 (d, 1H), 2.75 (m, 2H), 2.08 (m, 1H), 1.97 (m, 1H), 1.85 (m, 2H), 1.75 (m, 1H), 1.65 (m, 1H), 1.52 (m, 1H(, 1.35 (m, 1H)

[0083] Example 1-2: Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dioxalate I JPEG0007644873000004.jpg30107 5 mL of methanol was added to 100 mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) prepared in the above Preparation Example, and 76.9 mg (2 eq.) of oxalic acid dehydrate was added and stirred at room temperature for 1 day. After leaving at room temperature for 1 day until a solid was formed, the solid was washed to obtain the title compound (108 mg, 70% yield).

[0084] Molecular weight 508.31 1 H-NMR (500 MHz, DMSO): δ 8.40 (s, 1H). 7.69 (d, 1H), 7.48 (d, 1H), 4.31 (m, 2H), 3.45 (m, 1H), 3.15 (m, 1H), 2.77 (m, 2H), 2.01 (m, 1H), 1.90 (m, 3H), 1.78 (m, 1H), 1.58 (m, 1H), 1.45 (m, 1H), 1.35 (m, 1H)

[0085] Example 1-3: Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dioxalate II JPEG0007644873000005.jpg30107 5mL of acetonitrile was added to 100mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) prepared in the above Preparation Example, and 76.9mg (2eq.) of oxalic acid dihydrate was added and stirred at room temperature for 1 day. After leaving at room temperature for 1 day until a solid was formed, the solid was washed to obtain the title compound (140mg, yield 90%).

[0086] Molecular weight 508.31 1 H-NMR (500 MHz, DMSO): δ 8.40 (s, 1H). 7.69 (d, 1H), 7.48 (d, 1H), 4.32 (m, 2H), 3.36 (m, 1H), 3.11 (d, 1H), 2.77 (m, 2H), 1.92 (m, 1H), 1.89 (m, 3H), 1.77 (m, 1H), 1.58 (m, 1H), 1.47 (m, 1H), 1.35 (m, 1H)

[0087] Example 1-4: Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol hemi-oxalate JPEG0007644873000006.jpg30107 102.2 mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) prepared in the above Preparation Example was placed in a round-bottom flask and completely dissolved in 5 mL of methanol. 19.6 mg (0.5 eq.) of oxalic acid dihydrate was added to the flask in which the free base was dissolved and stirred for 2 hours. The mixture was left to dry at room temperature, and the title compound (84 mg, yield 74%) was obtained as a solid.

[0088] Molecular weight 373.26 1 H-NMR (500 MHz, DMSO): δ 8.40 (s, 1H). 7.69 (d, 1H), 7.47 (d, 1H), 4.30 (m, 2H), 3.22 (m, 1H), 3.03 (d, 1H), 2.60 (m, 2H), 2.02 (m, 1H), 1.88 (m, 2H), 1.80 (m, 1H), 1.69 (m, 1H), 1.50 (m, 1H), 1.38 (m, 1H), 1.29 (m, 1H)

[0089] Examples 1-5: Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol hemi-maleate JPEG0007644873000007.jpg30107 100 mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) prepared in the above Preparation Example was placed in a vial and completely dissolved in 2.5 mL of methanol. 20.4 mg (0.5 eq.) of maleic acid was added to the vial in which the free base was dissolved and stirred for 2 hours. After this, the methanol was blown off, acetonitrile was added, and the mixture was stirred for 4 hours, and then the mixture was left to dry at room temperature in an open container to obtain the title compound (88 mg, yield 74.5%) as a solid.

[0090] Molecular weight 386.28 1 H-NMR (500 MHz, DMSO): δ 8.40 (s, 1H). 7.68 (d, 1H), 7.49 (d, 1H), 6.01 (s, 1H), 5.07 (s, 1H), 4.30 (m, 2H), 3.17 (s, 1H), 2.98 (d, 1H), 2.62 (m, 2H), 1.98 (m, 1H), 1.80 (m, 3H), 1.69 (m, 1H), 1.45 (m, 1H), 1.29 (m, 2H)

[0091] Example 1-6: Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol palmitate JPEG0007644873000008.jpg2498100mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol (free base) prepared in the above Preparation Example was placed in a vial and completely dissolved in 4.8mL of ethanol. 78.7mg (1eq.) of palmitic acid was added to the vial in which the free base was dissolved and stirred for 2 hours. After this, the ethanol was blown off, ethyl acetate was added, and the mixture was stirred for 4 hours, and then dried at room temperature in an open container to obtain the title compound (154mg, yield 86.5%) as a solid.

[0092] Molecular weight 584.67 1 H-NMR (500 MHz, DMSO): δ 8.39 (s, 1H). 7.66 (d, 1H), 7.46 (d, 1H), 4.26 (m, 2H), 2.90 (m, 1H), 2.79 (d, 1H), 2.31 (m, 1H), 2.15 (m, 3H), 1.98 (m, 1H), 1.78 (m, 3H), 1.52 (m, 1H), 1.46 (m, 2H), 1.28 (m, 1H), 1.23 (m, 24H), 1.13 (m, 2H), 0.85 (m, 3H)

[0093] Comparative Example 1: Preparation of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride JPEG0007644873000009.jpg2479 Steps 1-1 to 1-4 were carried out in the same manner as in Preparation Example 1 to obtain the title compound (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol diHCl (26 mg, yield 81%).

[0094] Molecular weight 401.15 1 H NMR (500 MHz, MeOD): δ 9.67 (s, 1H). 8.02 (d, 1H), 7.82 (d, 1H), 4.62 (m, 2H), 3.60 (m, 1H), 3.28 (m, 1H), 2.99 (m, 2H), 2.25 (m, 2H), 2.08 (m, 2H), 1.99 (m, 1H), 1.78 (m, 2H), 1.54 (m, 1H)

[0095] Test Example 1: PRS enzyme activity inhibition experiment The IC value of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 for inhibiting PRS enzyme activity 50 The values ​​were measured as follows:

[0096] Specifically, the portion of the EPRS cDNA corresponding to the PRS was subcloned, and the resulting highly purified PRS protein was purified and used in the experiment. The reaction buffer (20 mM KPO4 (pH 7.4), 6 mM MgAc, 5 mM ATP, 400 mg / mL tRNA, 0.5 mM DTT, 20 mCi[ 3 The compound (1 μM) prepared in the above Example was added to [H]proline (1 mCi / mL) and reacted at 37°C for 5 to 10 minutes. The reaction was terminated by adding 5% TCA and using pre-dried 3M paper. Radioactivity was measured using a liquid scintillation counter. % inhibition and IC 50 Calculation of values ​​and analysis of results were performed using Microsoft Excel or Sigma Plot 8.0, and the result was that (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride showed a value of 74 nM.

[0097] Therefore, it can be seen that (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride inhibits PRS enzyme activity and can be used in a pharmaceutical composition for preventing or treating diseases caused by abnormal PRS activity.

[0098] Test Example 2: Differential scanning calorimetry and thermogravimetry of acid addition salts Differential scanning calorimetry and thermogravimetry were carried out on the acid addition salts prepared in Examples 1-1 to 1-6 and Comparative Example 1, and the results are shown in Figures 1 to 7, respectively. Differential scanning calorimetry was carried out using a differential scanning calorimeter (DSC3, manufactured by Mettler-Toledo) in a sealed fan under nitrogen gas while heating from 20°C to 350°C at a scan rate of 10°C / min. Thermogravimetric analysis (TGA) was carried out using a thermogravimetric analyzer (Q50, manufactured by TA Instrument USA) while heating from 20°C to 350°C at a scan rate of 10°C / min under nitrogen gas.

[0099] 1, it can be seen that the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 exhibits an endothermic onset temperature of 279.68° C. and a maximum endothermic peak at an endothermic temperature of 281.04° C. in differential scanning calorimetry. In addition, no residual solvent or moisture was found in thermogravimetric analysis.

[0100] Referring to FIG. 2, it can be seen that (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dioxalate I prepared in Example 1-2 exhibits an endothermic onset temperature of 210.42° C. and a maximum endothermic peak at an endothermic temperature of 213.66° C. in differential scanning calorimetry.

[0101] 3, it can be seen that (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dioxalate II prepared in Example 1-3 shows an endothermic onset temperature of 193.84° C. and a maximum endothermic peak at an endothermic temperature of 203.92° C. in differential scanning calorimetry. However, when the peak shown at 64° C. in FIG. 3 was observed using a hot stage microscope (HSM, BA310Pol, manufactured by Motic), it was determined that this peak was due to a residual solvent and not moisture.

[0102] 4, (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol hemi-oxalate prepared in Example 1-4 exhibits an endothermic onset temperature of 253.72° C. and a maximum endothermic peak at an endothermic temperature of 255.62° C. in differential scanning calorimetry. In addition, thermogravimetric analysis shows that the salt melts and decomposes as the temperature increases.

[0103] 5, it can be seen that the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol hemi-maleate prepared in Example 1-5 exhibits an endothermic onset temperature of 90.69° C. and a maximum endothermic peak at an endothermic temperature of 110.16° C. in differential scanning calorimetry. In addition, the thermogravimetric analysis shows that the salt melts and decomposes as the temperature increases.

[0104] 6, it can be seen that the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol palmitate prepared in Example 1-6 exhibits an endothermic onset temperature of 92.32° C. and a maximum endothermic peak at an endothermic temperature of 98.59° C. in differential scanning calorimetry. In addition, the thermogravimetric analysis shows that the compound melts and decomposes as the temperature increases.

[0105] 7, it can be seen that the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 1 shows two endothermic peaks (a first peak having an endothermic onset temperature of 233.09° C. and an endothermic temperature of the maximum endothermic peak of the peaks of 242.09° C., and a second peak having an endothermic onset temperature of 271.95° C. and an endothermic temperature of the maximum endothermic peak of 275.07° C.). In addition, no residual solvent or moisture was found in the thermogravimetric analysis.

[0106] Test Example 3: Hygroscopicity test The acid addition salts prepared in Examples 1-1 to 1-6 and Comparative Example 1 were subjected to a hygroscopicity test.

[0107] 40 mg of each acid addition salt sample was placed in a 10 mL beaker and then sealed in a glass desiccator containing a saturated aqueous solution of various salts at room temperature and under a constant relative humidity condition as shown in Table 1 below. After measuring the weight change after one day, the sample was transferred to another desiccator and the above experiment was repeated. For sorption analysis, the sample was transferred from a desiccator with low humidity to a desiccator with high humidity and the weight change was measured, and for desorption analysis, the sample was transferred from a desiccator with high humidity to a desiccator with low humidity and the weight change was measured. The results are shown in Table 2 below.

[0108] [Table 1]

[0109] [Table 2]

[0110] Referring to Table 2, the acid addition salts of the Examples not only show almost no weight change due to moisture even when the relative humidity increases, but also show significantly lower hygroscopicity at 93% RH compared to the dihydrochloride salt of Comparative Example 1.

[0111] Test Example 4: Stability Confirmation Test The acid addition salts prepared in some of the Examples and Comparative Examples were subjected to a stability test under high temperature and high humidity.

[0112] Specifically, the temperature stress test was performed by placing 50 mg of a sample in a vial, capping it, and storing it in a chamber at 60°C and a relative humidity of less than 10%, and measuring the impurity content using HPLC (Azura, KNAUER, P6.11L, DAD6.1L) after 1 week, 2 weeks, 4 weeks, and 8 weeks.

[0113] The moisture stress test was performed by placing 50 mg of sample in a 10 mL beaker, storing it in a 93% RH desiccator without a lid so that the sample could come into sufficient contact with moisture in the air, and measuring the impurity content using HPLC (Azura, KNAUER, P6.11L, DAD6.1L) after 1 week, 2 weeks, 4 weeks, and 8 weeks.

[0114] (1) Calculation of peak areas for main components using HPLC At designated time points after the start of the test, one vial or beaker was taken out for each time point (number of samples per test, n=1) and impurities were analyzed using HPLC under the conditions shown in Table 3 below. After that, the peak areas of all detected peaks were obtained by integration, and the relative peak areas (%) for the main component were calculated and shown as average values, and the results are shown in Table 4 below.

[0115] [Table 3]

[0116] [Table 4] (*However, the dihydrochloride salt will deliquesce within one week at 93% RH.)

[0117] As shown in Tables 3 and 4, the acid addition salts prepared in the examples do not show a significant decrease in the peak area of ​​the main component under high temperature and high humidity exposure conditions. Therefore, it can be confirmed that the crystal forms prepared in the examples exhibit excellent chemical stability by suppressing the increase of impurities regardless of the influence of temperature and humidity under harsh conditions.

[0118] (2) Differential scanning calorimetry For (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 and (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 1, each beaker was taken out from the desiccator in which the temperature stress test was being performed 1 week and 8 weeks after the start of the test, and differential scanning calorimetry was performed, and the results are shown in Figures 8 to 11, respectively. At this time, the differential scanning calorimetry was performed using a differential scanning calorimeter (DSC Q20, manufactured by TA Instruments) in a sealed fan under nitrogen purification while increasing the temperature from 20°C to 300°C at a scan rate of 10°C / min.

[0119] Specifically, Figures 8 and 9 show the results of differential scanning calorimetry of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 after 1 week and 8 weeks of stress testing against temperature, respectively.

[0120] 10 and 11 are diagrams showing the results of differential scanning calorimetry (DSC) analysis of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 1 after one week and eight weeks of stress testing against temperature, respectively.

[0121] 8 to 11, the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 1 exhibited a change in differential scanning calorimetry pattern with the first peak at the front becoming smaller over time under high temperature conditions, whereas the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 showed no change in differential scanning calorimetry pattern even after 8 weeks under high temperature conditions. This confirms that (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride has excellent chemical stability.

[0122] Example 2-1: Preparation of crystalline form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride using acetonitrile solvent 10 mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 was suspended in 1 mL of acetonitrile solvent and stirred at room temperature for 3 hours. The suspension was then left at room temperature for at least one day to evaporate the solvent. After crystallization, the crystals were separated by filtration under reduced pressure to obtain 8 mg of crystalline form I of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride.

[0123] Examples 2-2 to 2-9 Crystals of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride were obtained in the same manner as in Example 2-1, except that the solvents shown in Table 5 below were used instead of acetonitrile solvent in Example 2-1. The differential scanning calorimetry of the crystals thus prepared showed that they had the same crystalline form (crystal form I) as in Example 2-1.

[0124] Example 2-10 10 mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 was dissolved in 1 mL of methanol solvent and stirred at room temperature for 3 hours. The solution was then left at room temperature for at least one day to evaporate the solvent. After crystallization, crystals were obtained by filtration under reduced pressure. Differential scanning calorimetry of the crystals thus prepared showed that they had the same crystalline form (crystal form I) as that of Example 2-1.

[0125] Example 2-11 10 mg of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1 was dissolved in 0.8 mL of methanol solvent to prepare a solution. Then, 1 mL of acetonitrile solvent was added as an anti-solvent to the prepared solution, and the solution was allowed to stand at room temperature for one day or more. After the crystals were generated, the crystals were obtained by filtration under reduced pressure. The differential scanning calorimetry of the prepared crystals showed that they had the same crystalline form (crystal form I) as that of Example 2-1.

[0126] Examples 2-12 to 2-19 Crystals of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride were obtained in the same manner as in Example 2-11, except that the solvents shown in Table 5 below were used instead of acetonitrile as the non-solvent in Example 2-11. The differential scanning calorimetry of the crystals thus prepared showed that they had the same crystalline form (crystal form I) as in Example 2-1.

[0127] [Table 5]

[0128] Referring to Table 5, it can be seen that the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol produces only one crystal form even when the crystallization method and the solvent used are different. In particular, it can be seen that the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol produced under the methanol / acetone solvent condition of Example 2-12 is a single crystal.

[0129] Comparative Example 2: Preparation of a crystalline form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride using acetonitrile solvent Since the (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 1 was in a crystalline form, it was used as Comparative Example 2.

[0130] Test Example 5: X-ray powder diffraction analysis X-ray powder diffraction analysis was performed on the crystal forms prepared in Example 2-1 and Comparative Example 2, and the results are shown in Figures 1 and 2, respectively. At this time, the X-ray powder diffraction analysis was performed using an X-ray powder diffraction spectrometer (D8 Advance, manufactured by Bruker) at a voltage of 45 kV, a current of 40 mA, a divergence and scattering slit of 1°, a receiving slit of 0.2 mm, and a scanning speed of 3° / min (0.4 sec / 0.02° intervals) using a CuKα target in the diffraction angle (2θ) range of 5° to 35°.

[0131] Referring to FIG. 12, it can be seen that the crystalline form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 2-1 has peaks at diffraction angles (2θ) of 7.5°, 12.5°, 15.0°, 16.5°, 17.2°, 18.9°, 19.8°, 20.9°, 22.3°, 23.6°, 24.8°, 25.1°, 26.1°, 27.8°, 28.9°, 30.1°, 30.9°, 31.3° and 32.4° in the X-ray powder diffraction pattern.

[0132] Referring to FIG. 13, it can be seen that the crystalline form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 2 has peaks at diffraction angles (2θ) of 9.6°, 10.9°, 12.2°, 13.9°, 15.8°, 16.9°, 17.9°, 19.2°, 20.2°, 20.8°, 22.2°, 23.6°, 24.5°, 24.8°, 25.2°, 25.6°, 27.6°, 27.9°, 28.6°, 29.2°, 30.2° and 31.5° in the X-ray powder diffraction pattern.

[0133] In addition, referring to Figures 12 and 13, the crystal form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 2-1 and the crystal form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol dihydrochloride prepared in Comparative Example 2 are different from each other.

[0134] Test Example 6: Differential scanning calorimetry of crystal form The crystalline form prepared in Example 2-1 was subjected to differential scanning calorimetry, and the results are shown in Figure 14. The differential scanning calorimetry was carried out using a differential scanning calorimeter (DSC Q20, manufactured by TA Instruments) in a sealed fan under nitrogen purification while increasing the temperature from 20°C to 300°C at a scan rate of 10°C / min.

[0135] 14, the crystal form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 2-1 shows an endothermic onset temperature of 279.68±3° C. and a maximum endothermic peak at an endothermic temperature of 281.04±3° C. in differential scanning calorimetry. Therefore, the crystal form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 2-1 not only has a higher endothermic onset temperature than the crystal form of the dihydrochloride, but also has a higher endothermic temperature at which the maximum endothermic peak occurs, and thus has improved specific heat stability compared to the crystal form of the dihydrochloride.

[0136] The differential scanning calorimetry analysis result of the crystalline form of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 2-1 was the same as the differential scanning calorimetry analysis result of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride prepared in Example 1-1. Therefore, it can be seen that the monohydrochloride prepared in Example 1-1 is also a crystalline form, and its crystalline form is the same as the crystalline form (crystalline form I) of the monohydrochloride prepared in Example 2-1.

Claims

1. (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol monohydrochloride.

2. In differential scanning calorimetry (DSC) analysis, the endothermic onset temperature is 279.68±3°C and the maximum endothermic peak is at an endothermic temperature of 281.04±3°C. The monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 1.

3. An acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, The acid addition salt is an oxalate, a maleate, or a palmitate. Acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

4. The acid addition salt is a dioxalate salt. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 3.

5. The dioxalate salt has an endothermic onset temperature of 210.42±3° C. and a maximum endothermic peak at an endothermic temperature of 213.66±3° C. in differential scanning calorimetry. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 4.

6. The dioxalate salt has an endothermic onset temperature of 193.84±3° C. and a maximum endothermic peak at an endothermic temperature of 203.92±3° C. in differential scanning calorimetry. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 4.

7. The acid addition salt is a hemi-oxalate salt. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 3.

8. The hemi-oxalate salt exhibits an endothermic onset temperature of 253.72±3° C. and a maximum endothermic peak at an endothermic temperature of 255.62±3° C. in differential scanning calorimetry. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 7.

9. The acid addition salt is a hemi-maleate salt. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 3.

10. The hemi-maleate salt exhibits an endothermic onset temperature of 90.69±3° C. and a maximum endothermic peak at an endothermic temperature of 110.16±3° C. in differential scanning calorimetry.

10. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 9.

11. The acid addition salt is a palmitate salt. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 3.

12. The palmitate salt has an endothermic onset temperature of 92.32±3° C. and a maximum endothermic peak at an endothermic temperature of 98.59±3° C. in differential scanning calorimetry. The acid addition salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 11.

13. having peaks at diffraction angles (2θ±0.2°) of 15.0°, 17.2°, 19.8°, 23.6° and 27.8° in an X-ray powder diffraction pattern; A crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

14. In differential scanning calorimetry, the endothermic onset temperature is 279.68±3°C and the maximum endothermic peak is at an endothermic temperature of 281.04±3°C. The crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol according to claim 13.

15. Monohydrochloride of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, oxalate of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, maleate of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol 2. A pharmaceutical composition for preventing or treating cancer, an inflammatory disease, an autoimmune disease, or a fibrotic disease, comprising one or more selected from the group consisting of a crystalline form of a salt, a palmitate salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol, and a crystalline form of the monohydrochloride salt of (2R,3S)-2-(3-(4,5-dichloro-1H-benzo[d]imidazol-1-yl)propyl)piperidin-3-ol.

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