Novel crystal of n-[4-(4-carbamoyl benzamide)benzene-1-sulfonyl]-d-γ-glutamyl-(4S)-4-amino-l-prolyl-l-leucyl-n-(5-amino-5-oxopentyl)-n2-methyl-l-α-glutamine, and method for producing the same

Abstract

To provide a novel crystal of a compound for inhibiting matrix metalloprotease 2, and a method for producing the crystal.SOLUTION: The problem described above is solved by providing a crystal of a DMF solvate of a compound represented by the following formula [I], as well as a method for producing the crystal.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a compound comprising N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N 2 The present invention relates to a crystal of α-methyl-L-α-glutamine DMF solvate and a related method. [Background technology]

[0002] Matrix metalloproteinases (hereinafter referred to as "MMPs") are endopeptidases with zinc at their active center, and 24 genes are known to encode them. MMPs degrade extracellular matrices such as collagen and gelatin, and are therefore involved not only in physiological phenomena such as bone remodeling and wound healing, but also in pathological processes such as inflammation and cancer progression (see Non-Patent Document 1).

[0003] To date, clinical trials of several MMP inhibitors have been conducted, focusing on the anti-cancer effects of MMP inhibition. However, these have been abandoned due to side effects such as skeletal muscle pain, which are thought to be caused by the relatively non-selective inhibitory effect on various MMP subtypes, and the possibility of promoting cancer metastasis (see Non-Patent Documents 2 and 3).

[0004] Activation of matrix metalloproteinase 2 (hereinafter referred to as "MMP2") has been reported to play an important role in cancer cell invasion and metastasis. Cancer cell invasion and metastasis are important factors affecting the prognosis of malignant tumors, and inhibiting MMP2 activity could be an effective therapeutic approach for cancer control. It has been reported that cancer growth is suppressed in MMP2 gene-deficient animals, indicating that MMP2 plays an important role in cancer growth (see Non-Patent Document 4). Furthermore, MMP2 has been reported to be associated with the progression of various cancers, including breast cancer, pancreatic cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, brain tumor, gastric cancer, hepatocellular carcinoma, head and neck cancer, melanoma, uterine cancer, esophageal cancer, renal cell carcinoma, lung cancer, and glioma (see Non-Patent Documents 5 and 6). On the other hand, MMP2 has also been reported to be involved in the pathogenesis of non-neoplastic diseases.

[0005] It has been reported that in chronic kidney disease, MMP2 induces epithelial-mesenchymal transition of the tubules by changing the structure of the tubular basement membrane, leading to tubular atrophy, fibrosis, and impaired renal function (see Non-Patent Document 7). Furthermore, elevated MMP2 concentrations have been observed in the blood of patients with chronic kidney disease (see Non-Patent Documents 8 and 9). It has also been reported that renal fibrosis induced by unilateral ureteral ligation is suppressed in MMP2 gene-deficient animals (see Non-Patent Documents 10 and 11). Therefore, inhibiting MMP2 activity may be an effective therapeutic approach for controlling the progression of chronic kidney disease.

[0006] In idiopathic pulmonary fibrosis, elevated MMP2 expression has been observed in alveolar epithelial cells, fibroblasts, and macrophages, and in particular, elevated MMP2 expression has been observed in alveolar lavage fluid in patients with rapidly progressing idiopathic pulmonary fibrosis (see Non-Patent Documents 12 and 13). It has also been reported that non-selective MMP inhibitors reduce collagen content in the lungs of mice with bleomycin-induced pulmonary fibrosis (see Non-Patent Document 14) and suppress TGFβ-induced transformation of pulmonary parenchymal fibroblasts (see Non-Patent Document 15). Therefore, inhibiting MMP2 activity may be an effective therapeutic approach for controlling the progression of idiopathic pulmonary fibrosis.

[0007] Furthermore, a relationship between MMP2 and non-tumor diseases such as multiple sclerosis, cerebral infarction, arteriosclerosis, abdominal aortic aneurysm, peritoneal sclerosis, myocardial infarction, acute kidney injury, diabetic nephropathy, nephrosclerosis, glomerulonephritis, polycystic kidney disease, polycystic liver, alcoholic liver disease, non-alcoholic fatty liver disease, cholestatic liver injury, chronic obstructive pulmonary disease, interstitial pneumonia, diabetic retinopathy, age-related macular degeneration, Sjögren's syndrome, meningitis, muscular dystrophy, scleroderma, inflammatory bowel disease, and tuberculosis has been suggested (see Non-Patent Document 16).

[0008] Based on these findings, finding a means for selectively inhibiting MMP2 is a highly feasible approach to establishing effective treatments for diseases in which MMP2 is involved.

[0009] There have been reports of low molecular weight compounds that have MMP2 inhibitory activity, including compounds incorporating hydroxamic acid or carboxylic acid as zinc chelators. However, none of these low molecular weight compounds are known to exhibit selective MMP2 inhibitory activity (see, for example, Non-Patent Documents 17 and 18).

[0010] Here, examples of compounds exhibiting selective MMP2 inhibitory activity include the peptide compound "β-amyloid precursor protein (APP-IP, IIe-Ser-Tyr-Gly-Asn-Asp-Ala-Leu-Met-Pro)" (see Non-Patent Document 19) and the peptide compound substituted with a substituted phenylsulfonamide, "N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N ... 2 -methyl-L-α-glutamine" (hereinafter, also referred to as compound [I]) has been reported (see Patent Documents 1 and 2).

[0011] [ka]

[0012] It has been reported that compound [I] is also useful in the prevention or treatment of pulmonary inflammation and fibrosis (see Patent Document 2).

[0013] In the manufacture of pharmaceuticals, it is generally desired that the physical properties of the compound be such that it is easy to handle on an industrial scale and has excellent storage stability. However, there has been no report yet on the above-mentioned compound [I] in a form that is easy to handle and has excellent storage stability, namely, a crystal form. [Prior art documents] [Patent documents]

[0014] [Patent Document 1] WO2021 / 090959 [Patent Document 2] WO2023 / 204170 [Non-patent literature]

[0015] [Non-Patent Document 1] HJ Ra and WC Parks Matrix Biol., 2007, 26(8), 587-596. [Non-licensed document 2] AH Drummond et al. Ann NY Acad Sci., 1999, 878, 228-235. [Non-licensed document 3] AD Baxter et al. Bioorg Med Chem Lett., 2001, 11, 1465-1468.

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

Non-licensed literature 9

Non-licensed literature 10

Non-licensed Document 11

Non-licensed Document 12

[0016] The crystals of the substance are easily purified when separated from the slurry, and can also be purified by a simple method called recrystallization. In this regard, a white powder of compound [I] can be obtained by the method described in Example 1 of Patent Document 1, but the white powder obtained using this method was amorphous. Therefore, it was difficult to purify compound [I] by crystallization or recrystallization.

[0017] An object of the present invention is to provide a method for the preparation of a compound represented by the formula [I], which has an excellent MMP2 inhibitory activity, and which is characterized by the following: 2 The present invention aims to provide a novel crystal of α-methyl-L-α-glutamine (compound [I]). [Means for solving the problem]

[0018] The present inventors have conducted extensive research to achieve the above object, and have surprisingly discovered that by selecting and using DMF from among the vast number of solvents available, it is possible to obtain crystals of the DMF solvate of compound [I].

[0019] Furthermore, the inventors have discovered that compound [I] can be easily produced with the purity required for pharmaceuticals by converting the crystals into the free form of compound [I] using a conventional method.

[0020] The present invention will be described in detail below. (1) One aspect of the present invention is This is a crystal of a DMF solvate of the compound represented by the following formula [I].

[0021] [ka]

[0022] (2) Another aspect of the present invention is The crystal described in (1) has the following physical properties (a) to (b): (a) In powder X-ray diffraction (Cu-Kα), it has peaks at 2θ = 4.0 degrees, 8.0 degrees, 15.6 degrees, 17.8 degrees, and 21.6 degrees; and (b) In differential thermal analysis (DTA), it has an endothermic peak at 150 to 180°C. (3) Another aspect of the present invention is The following formula [I]:

[0023] [ka]

[0024] and crystallizing a DMF solvate of the compound represented by formula [I] from the mixture obtained in the step. (4) Another aspect of the present invention is The following formula [I]:

[0025] [ka]

[0026] and removing the solvent. (5) Another aspect of the present invention is The crystal according to (2) further has the following physical property (c): (c) Infrared absorption spectrum (KBr method) shows a characteristic absorption band at 3358 cm -1 , 1741cm -1 , 1683cm -1 , 1527cm -1 , 1172cm -1 , and 1010 cm -1 is located. (6) Another aspect of the present invention is The method for producing a crystal according to (3), wherein the crystal further has the following physical property (c): (c) Infrared absorption spectrum (KBr method) shows a characteristic absorption band at 3358 cm -1 , 1741cm -1 , 1683cm -1 , 1527cm -1 , 1172cm -1 , and 1010 cm -1 is located. (7) Another aspect of the present invention is The pharmaceutical composition contains the compound represented by the formula [I] obtained by the production method described in (4) above. (8) Another aspect of the present invention is The MMP2 inhibitor contains the compound represented by formula [I] obtained by the production method described in (4) above. (9) Another aspect of the present invention is The present invention provides a preventive or therapeutic agent for cancer disease or organ fibrosis, or a symptom associated with cancer disease or organ fibrosis, which contains the compound represented by formula [I] obtained by the production method described in (4) above. (10) Another aspect of the present invention is The present invention provides a preventive or therapeutic drug for pulmonary inflammation and fibrosis, which contains the compound represented by formula [I] obtained by the production method described in (4) above. (11) Another aspect of the present invention is The preventive or therapeutic drug according to (10), wherein the pulmonary inflammation and fibrosis is one or more diseases selected from the group consisting of acute respiratory distress syndrome, idiopathic pulmonary fibrosis, interstitial pneumonia, bacterial pneumonia, and viral pneumonia. (12) Another aspect of the present invention is The preventive or therapeutic drug according to (10) or (11), wherein the pulmonary inflammation and fibrosis is acute respiratory distress syndrome. (13) Another aspect of the present invention is The preventive or therapeutic drug according to (10) or (11), wherein the pulmonary inflammation and fibrosis is idiopathic pulmonary fibrosis. [Effects of the Invention]

[0027] The present invention provides a crystalline DMF solvate of compound [I], which has excellent MMP2 inhibitory activity. The crystalline DMF solvate is stable at temperatures around room temperature and has excellent storage stability.

[0028] Furthermore, since many impurities remain in the liquid phase of the slurry during crystallization, a certain degree of purification effect is expected when the crystals are separated from the slurry. The DMF solvate can also be recrystallized. Furthermore, by treating the crystals obtained by crystallization or recrystallization with a solvent, purified compound [I] (preferably in amorphous form) can be easily obtained. In this way, a novel method for producing compound [I] that stably obtains compound [I] with consistent quality has been provided, thereby enabling the convenient production of highly pure compound [I].

[0029] Furthermore, it would be completely unpredictable for a person skilled in the art that DMF, among the vast number of solvents available, would produce crystals of compound [I]. Furthermore, as far as the applicant knows, it is extremely rare for DMF to produce a solvate. [Brief explanation of the drawings]

[0030] [Figure 1] 1 shows the powder X-ray diffraction pattern of a crystal of the DMF solvate of N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N2-methyl-L-α-glutamine (compound [I]). [Figure 2] 1 shows the thermogravimetry / differential thermal analysis curves of a crystal of a DMF solvate of N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N2-methyl-L-α-glutamine (compound [I]). [Figure 3] 1 shows an infrared absorption spectrum (KBr method) of a crystal of a DMF solvate of N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N2-methyl-L-α-glutamine (compound [I]). [Figure 4]1H nuclear magnetic resonance analysis chart of a crystal of the DMF solvate of N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N2-methyl-L-α-glutamine (compound [I]). [Figure 5] 1H nuclear magnetic resonance analysis chart of amorphous N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N2-methyl-L-α-glutamine (compound [I]). [Figure 6] 1 shows the amorphous powder X-ray diffraction pattern of N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N2-methyl-L-α-glutamine (compound [I]). DETAILED DESCRIPTION OF THE INVENTION

[0031] Hereinafter, embodiments for carrying out the present invention will be specifically described. The DMF solvate of compound [I] of the present invention (hereinafter sometimes referred to as "the compound of the present invention") has the chemical structure shown in the following formula [I-1], in which DMF is attracted to and bound to the compound [I] to form a solvate.

[0032] [ka]

[0033] The crystals of the DMF solvate (hereinafter, sometimes referred to as "the crystals of the present invention") can be obtained with good reproducibility as single crystals having a certain quality as described above, can be stably supplied as crystals of drug substances used in the production of pharmaceuticals, and have excellent storage stability.

[0034] In the DMF solvate, the molar ratio of compound [I] to DMF is not limited. In one embodiment, the crystal of the present invention has the following physical properties (a) to (b): (a) In powder X-ray diffraction (Cu-Kα), it has peaks at 2θ=4.0 degrees, 8.0 degrees, 15.6 degrees, 17.8 degrees, and 21.6 degrees; and (b) In differential thermal analysis (DTA), it has an endothermic peak at 150 to 180°C.

[0035] In one embodiment, the crystal of the present invention further has the following physical property (c): (c) Infrared absorption spectrum (KBr method) shows a characteristic absorption band at 3358 cm -1 , 1741cm -1 , 1683cm -1 , 1527cm -1 , 1172cm -1 , and 1010 cm -1 is located.

[0036] The powder X-ray diffraction pattern of the crystal of the present invention is shown in FIG. 1, the thermogravimetry / differential thermal analysis curve is shown in FIG. 2, and the infrared absorption spectrum (KBr method) is shown in FIG. Here, in differential thermal analysis (DTA), an endothermic peak was observed in the range of 150 to 180°C, as shown in the chart in Figure 2. In addition, a broad endothermic peak appeared in the range of 100 to 150°C, and an endothermic peak was also observed around 180°C.

[0037] Furthermore, thermogravimetry (TG) analysis revealed that weight loss occurred in the ranges of 100 to 150°C and 150 to 180°C. Note that characteristic peaks in powder X-ray diffraction, thermogravimetry / differential thermal analysis (TG / DTA), and infrared absorption spectrum may vary depending on the measurement conditions, and therefore, these peaks of the compound of the present invention may be inaccurate or unclear.

[0038] As can be seen from Figures 1 to 3, the crystals of the DMF solvate of compound [I] produced by the production method of the present invention can basically have high purity. The purity of the crystals is desirably high, and preferably they are substantially free of other crystalline forms.

[0039] Furthermore, as will be shown in the Examples below, the crystals of the DMF solvate of compound [I] produced by the production method of the present invention can be reproducibly obtained as single crystals having a consistent quality, and can be stably supplied as crystals used in the production of pharmaceuticals, and have excellent physicochemical properties including excellent storage stability.

[0040] Next, a method for producing the crystal of the present invention will be described. The crystal of the present invention can be produced, for example, by the following method. A method for producing the crystal of the present invention, comprising the steps of mixing compound [I] with DMF and crystallizing the compound from the mixture obtained in the mixing step.

[0041] In one embodiment, the mixing step comprises mixing compound [I] with a first solvent containing DMF. The mixing step may comprise dissolving compound [I] in the first solvent. The first solvent may be heated for dissolution. The crystallization step may comprise cooling the mixture or solution obtained in the mixing step and / or mixing the mixture or solution with a predetermined second solvent. Here, the second solvent may also contain DMF.

[0042] In another embodiment, the mixing step comprises mixing compound [I] with a first solvent not containing DMF. The mixing step may comprise dissolving compound [I] in the first solvent. The first solvent may be heated for dissolution. The mixing step then comprises mixing a mixture or solution containing the first solvent with a second solvent containing DMF. The crystallization step is then carried out. In the crystallization step, the mixture or solution obtained in the mixing step may be cooled.

[0043] In the above mixing, either of the two components to be mixed may be added to the other. Compound [I], which is the starting material for the method for producing the crystal of the present invention, may be obtained from its pharmaceutically acceptable salt. In this case, the method of the present invention further comprises a step of treating the salt with an acid or base to remove counterions (desalting) to produce compound [I]. This step may be carried out in the first solvent, in which case the desalting step and the mixing step are carried out substantially simultaneously.

[0044] The crystals of the present invention obtained in the crystallization step can be collected by filtration, separated from the solvent by distillation under reduced pressure or centrifugation, etc., and then dried as necessary to obtain the crystals of the present invention. The above crystallization step may be repeated not only once but twice or more times, but is usually carried out only once.

[0045] The crystallization step can be carried out using only the first solvent, or a mixed solvent of the first solvent and the second solvent. The first and second solvents are not limited, but may be, for example, alcohols such as methanol, ethanol, and 2-propanol, DMF, acetonitrile, or a mixture of two or more thereof, and preferably DMF, acetonitrile, or a mixture thereof. The mixing ratio of these solvents may be appropriately changed.

[0046] Furthermore, the obtained crystal of the present invention can be recrystallized. Examples of solvents used for recrystallization are the same as those exemplified above for the first or second solvent. The amount of solvent used for crystallization or recrystallization is not particularly limited, but is preferably 1 to 50 times the amount of compound [I].

[0047] The temperature at which compound [I] or the crystals of the present invention are dissolved in a solvent used for crystallization or recrystallization is not particularly limited, but is usually from room temperature to 100°C, preferably from room temperature to 40°C. The temperature at which the crystals of the present invention are precipitated in crystallization or recrystallization is not particularly limited, but is usually from ice-cold temperature to 50° C. Preferably, it is from ice-cold temperature to room temperature, more preferably room temperature.

[0048] The time for precipitating the crystals of the present invention is not particularly limited, but is usually 1 hour to 7 days, preferably 1 hour to 3 days, and more preferably 24 hours. The cooling time for the crystallization step or recrystallization is not particularly limited as long as it is 10 seconds or more, but is usually 10 minutes to 24 hours, preferably 30 minutes to 5 hours.

[0049] Seed crystals can be used for crystallization or recrystallization. Seed crystals can be obtained by methods well known to those skilled in the art, such as scraping the wall of a vessel containing a solution for crystallization with a spatula, or by a solvent evaporation method in which crystals are obtained by concentrating a compound solution by slowly evaporating a solvent.

[0050] In the above-mentioned production method, compound [I] or a pharmaceutically acceptable salt thereof is amorphous or crystalline before being dissolved in the first solvent.

[0051] The temperature at which the crystals of the present invention are dried is not particularly limited, but is usually from ice-cold temperature to 100°C, preferably from room temperature to 50°C, and more preferably 40°C. The time for drying the crystals of the present invention is not particularly limited, but is usually 1 hour to 7 days, preferably 1 hour to 3 days, and more preferably 4 hours.

[0052] The compound of the present invention, namely, compound [I] (N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N 2 -methyl-L-α-glutamine) is described below.

[0053] The compounds of the present invention may be in amorphous form. The amorphous powder X-ray diffraction pattern of compound [I] is shown in FIG. Next, a method for producing compound [I] from the crystal of the present invention will be described.

[0054] Compound [I] can be produced by known methods, for example, by the following method. A method for producing compound [I], comprising mixing the present crystal with a solvent and removing the solvent.

[0055] In one embodiment, the mixing step involves mixing the crystals of the present invention with a predetermined first solvent. The crystals of the present invention may be dissolved in the first solvent. To this end, the mixture may be heated if necessary. The solution may then be freeze-dried to remove the solvent. In some cases, the solvent may be partially evaporated before freeze-drying.

[0056] In another embodiment, in the mixing step, the crystal of the present invention is dissolved in a predetermined first solvent, if necessary by heating, and then compound [I] is precipitated. For precipitation, a second solvent may or may not be mixed with the mixed solution. The liquid obtained by precipitating compound [I] is subjected to the removal step. In the removal step, the precipitate is collected by filtration, and if necessary, separated from the solvent by distillation under reduced pressure, centrifugation, or the like, and then dried to produce compound [I].

[0057] Furthermore, the compound [I] obtained by the above production method may be in an amorphous form. In the mixing step of the above production method, the first solvent may be added to the compound [I], or the compound [I] may be added to the first solvent.

[0058] In the above manufacturing method, in the step of mixing the second solvent, the second solvent may be added to a mixed solution containing the first solution, or a mixed solution containing the first solution may be added to the second solvent.

[0059] Specific examples of the first solvent include ethanol and / or water, and preferably ethanol and water. These solvents may be used in combination.

[0060] A specific example of the second solvent is ethanol. Furthermore, the mixing ratio between the first solvents and between the first solvent and the second solvent can be changed as appropriate.

[0061] The amount of the first solvent is not particularly limited, but is preferably 1 to 10 times the amount of the crystal of the present invention. The temperature at which the crystals of the present invention are dissolved in the first solvent is not particularly limited, but is usually from ice-cold temperature to 50° C. Preferably, it is from ice-cold temperature to room temperature, more preferably room temperature.

[0062] The amount of the second solvent is not particularly limited, but is preferably 1 to 100 times the amount of the crystal of the present invention. The temperature at which the second solvent is added and compound [I] is precipitated is not particularly limited, but is usually from ice-cold temperature to 50° C. Preferably, the temperature is from ice-cold temperature to room temperature, more preferably room temperature.

[0063] The time for adding the second solvent and precipitating compound [I] is not particularly limited, but is usually 1 hour to 7 days, preferably 1 hour to 3 days, one more preferred time being 2 hours, and another more preferred time being 2 days.

[0064] The temperature at which compound [I] is dried is not particularly limited, but is usually from ice-cold temperature to 50° C. Preferably, it is from room temperature to 50° C., more preferably room temperature. The time for drying compound [I] is not particularly limited, but is usually 1 hour to 7 days, preferably 1 hour to 3 days, and more preferably 1 hour to 24 hours.

[0065] Compound [I] of the present invention may form various solvates (excluding DMF solvates), including hydrates, and these solvates are also included within the scope of compound [I] of the present invention. The "solvate" as used herein is not particularly limited as long as it forms a solvate with compound [I], and examples thereof include hydrates and alcohol solvates such as ethanol solvates.

[0066] The compound [I] of the present invention is a single compound having a specific configuration due to an asymmetric center, and it exists as various optical isomers. The various optical isomers can be separated from the compound [I] by methods well known to those skilled in the art, such as fractional crystallization, or the compound [I] can be obtained by organic chemistry techniques well known for this purpose.

[0067] Compound [I] of the present invention may absorb moisture upon exposure to the atmosphere or upon recrystallization, and may become adsorbed water or form a hydrate. Compound [I] of the present invention also includes such hydrates.

[0068] "Matrix metalloproteinase 2 (MMP2)" is a type of endopeptidase that has zinc at its active center. As mentioned above, MMP2 degrades extracellular matrices such as collagen and gelatin, and is therefore involved in cell invasion, migration, metastasis, and other conditions, such as cancer and organ fibrosis.

[0069] As mentioned above, inhibition of MMP2 is also useful in the prevention or treatment of pulmonary inflammation and fibrosis. Therefore, by inhibiting MMP2, it is possible to prevent or treat cancer diseases and organ fibrosis, symptoms associated with cancer diseases and organ fibrosis, and lung inflammation and fibrosis.

[0070] The compound [I] of the present invention has an inhibitory effect on MMP2, and therefore can be used as an MMP2 inhibitor or an active ingredient in a preventive or therapeutic drug for cancer diseases and organ fibrosis.

[0071] Compound [I] can also be used as an active ingredient in drugs for preventing or treating symptoms associated with cancer diseases and organ fibrosis. Furthermore, compound [I] can also be used as an active ingredient in drugs for the prevention or treatment of pulmonary inflammation and fibrosis.

[0072] As used herein, "cancer diseases" include breast cancer, pancreatic cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, brain tumor, gastric cancer, hepatocellular carcinoma, head and neck cancer, melanoma, uterine cancer, esophageal cancer, renal cell carcinoma, lung cancer, glioma, etc. Furthermore, "symptoms associated with cancer diseases" include pain, weight loss, and paraneoplastic syndromes associated with an increase in neoplastic cells and tumor growth.

[0073] As used herein, "organ fibrosis" includes chronic kidney disease, interstitial pneumonia, idiopathic pulmonary fibrosis, etc. Furthermore, "symptoms associated with organ fibrosis" include proteinuria and renal dysfunction in chronic kidney disease, and exertional dyspnea and dry cough in interstitial pneumonia and idiopathic pulmonary fibrosis.

[0074] As used herein, the term "a prophylactic or therapeutic agent for pulmonary inflammation and fibrosis" refers to a pharmaceutical agent that prevents or treats pulmonary inflammation and fibrosis. As used herein, "pulmonary inflammation and fibrosis" includes idiopathic interstitial pneumonia, idiopathic pulmonary fibrosis, pneumoconiosis, acute lung injury, acute respiratory distress syndrome, pulmonary edema, bacterial pneumonia, viral pneumonia, atypical pneumonia, asthma, eosinophilic pneumonia, hypersensitivity pneumonitis, sarcoidosis, ANCA-associated pulmonary disease, sarcoidosis, chronic obstructive pulmonary disease (COPD), and the like.

[0075] As used herein, "pulmonary inflammation" refers to acute inflammation of the alveolar cavity, alveolar epithelium, and alveolar septa caused by bacterial infection, viral infection, or the like. As used herein, "pulmonary fibrosis" refers to chronic fibrotic lesions caused by alveolar damage due to various causes, followed by abnormal repair and extracellular matrix deposition, and is a disease observed in the final stage of pulmonary inflammatory responses.

[0076] As used herein, "acute respiratory distress syndrome" refers to a pulmonary disease that rapidly develops hyperpermeability pulmonary edema due to direct or indirect injury, resulting in severe hypoxemia. As used herein, "idiopathic pulmonary fibrosis" refers to a lung disease characterized by chronic and progressive fibrosis and honeycombing, which is the most frequent type of idiopathic interstitial pneumonia.

[0077] As used herein, "idiopathic interstitial pneumonia" includes idiopathic pulmonary fibrosis, nonspecific interstitial pneumonia, desquamative interstitial pneumonia, interstitial lung disease associated with respiratory bronchiolitis, idiopathic organizing pneumonia, acute interstitial pneumonia, lymphocytic interstitial pneumonia, and the like.

[0078] As used herein, "bacterial pneumonia" includes pneumonia caused by gram-positive cocci such as Streptococcus pneumoniae and Staphylococcus aureus, and pneumonia caused by gram-negative bacilli such as Pseudomonas aeruginosa and Klebsiella pneumoniae.

[0079] As used herein, "viral pneumonia" includes pneumonia caused by respiratory viruses such as influenza virus, RS virus, and adenovirus, and pneumonia caused by systemic viruses such as measles virus and rubella virus.

[0080] "Prevention" means administering the medicament of the present invention to a patient at risk of developing a disease or disorder such as pulmonary inflammation and fibrosis before the disease develops. "Treatment" refers to administering the medicament of the present invention to a patient who has already developed a disease or disorder such as pulmonary inflammation and fibrosis. Such treatment includes symptomatic treatment to relieve symptoms resulting from the disease. It also includes treatment to reverse or partially reverse the disease, or treatment to halt or slow the progression of the disease.

[0081] The inhibitory activity of the compound [I] of the present invention against MMP2 can be measured by known methods.

[0082] Therefore, a person skilled in the art can measure the inhibitory activity of compound [I] against MMP2 by using the above-mentioned known measurement method. Similarly, the inhibitory activity of compound [I] against various MMP subtypes can also be measured.

[0083] In one aspect, the present invention is a pharmaceutical comprising compound [I]. The pharmaceutical of the present invention may be administered with compound [I] alone or together with a pharmaceutically or pharmaceutically acceptable excipient.

[0084] The pharmaceutical agent of the present invention can be administered orally or parenterally. The preferred method of administration is oral administration. Parenteral administration includes intravenous administration, nasal administration, transdermal administration, subcutaneous administration, intramuscular administration, and sublingual administration, with intravenous administration, subcutaneous administration, and transdermal administration being preferred.

[0085] The pharmaceutical of the present invention may be in any form of a solid composition, a liquid composition, or other composition, and the most suitable one can be selected according to the need.

[0086] The medicine of the present invention can be prepared by appropriately using the above-mentioned compound [I] and pharmaceutically acceptable additives, such as known carriers and diluents, and formulating them into an appropriate pharmaceutical composition form by a conventional method.

[0087] Specifically, the pharmaceutical can be used as an oral preparation such as a tablet, powder, dispersing agent, fine granules, granules, liquid preparation, coated tablet, capsule, syrup, jelly, troche, inhalant, etc., and as a parenteral preparation such as an injection, infusion, drip infusion, implant, transdermal preparation, transmucosal preparation, nasal preparation, enteral preparation, suppository, patch, etc. The parenteral preparation may be a microsphere preparation.

[0088] Preferred oral dosage forms include tablets, powders, fine granules, granules, coated tablets, capsules, syrups, lozenges, inhalants, etc. Preferred parenteral dosage forms include injections, infusions, etc.

[0089] The pharmaceutical of the present invention can also be formulated by forming an inclusion compound between compound [1] and α-, β- or γ-cyclodextrin, methylated cyclodextrin, or the like.

[0090] The pharmaceutical of the present invention can be a single preparation (combined preparation) of compounds that can be used in combination with compound [I], or two or more preparations (combined preparations) obtained by separately formulating the compounds. When these compounds are formulated separately to form two or more formulations, the individual formulations can be administered simultaneously or at a fixed time interval. In this case, it does not matter which formulation is administered first. The two or more formulations can also be administered at different times per day. Furthermore, the two or more formulations can also be administered by different routes.

[0091] When these compounds are formulated separately to form two different preparations, they may be administered simultaneously or at a very short interval. For example, it is preferable to state in documents such as package inserts and sales pamphlets of commercially available pharmaceuticals that they are to be used in combination.

[0092] It is also preferable to formulate these active ingredients separately into a kit consisting of two different formulations.

[0093] When compound [I] of the present invention is used as an MMP2 inhibitor or the like, the administration route is not particularly limited, and compound [I] can be administered orally or parenterally as it is. Alternatively, compound [I] may be administered orally or parenterally as a formulation containing compound [I] as an active ingredient.

[0094] In addition, when compound [I] is used as a drug for preventing or treating cancer diseases and organ fibrosis, symptoms associated with cancer diseases and organ fibrosis, and lung inflammation and fibrosis, compound [I] can be administered orally or parenterally as it is, or as a formulation containing compound [I] as an active ingredient.

[0095] When the pharmaceutical of the present invention is in the form of an oral preparation, other known additives, such as vitamins, amino acids, herbal medicines, natural products, excipients, pH adjusters, cooling agents, suspending agents, thickening agents, solubilizing agents, disintegrants, binders, lubricants, antioxidants, coating agents, colorants, flavoring agents, surfactants, plasticizers, fragrances, stabilizers, etc., can be mixed as needed within qualitative and quantitative ranges that do not impair the effects of the invention.

[0096] Examples of excipients include lactose, starch, crystalline cellulose, mannitol, maltose, calcium hydrogen phosphate, light anhydrous silicic acid, and calcium carbonate; examples of disintegrants include starch and calcium carboxymethylcellulose; examples of binders include starch, polyvinylpyrrolidone, hydroxypropyl cellulose, ethyl cellulose, carboxymethyl cellulose, and gum arabic; examples of lubricants include magnesium stearate, talc, and hydrogenated oil; and examples of stabilizers include lactose, mannitol, maltose, polysorbates, macrogols, and polyoxyethylene hydrogenated castor oil.

[0097] Furthermore, when the pharmaceutical of the present invention is in the form of a parenteral preparation, other known additives, such as injection solutions (including aqueous solutions and oily solutions), stabilizers, solubilizing agents, buffers, soothing agents, suspending agents, emulsifiers, preservatives, etc., can be mixed as needed, specifically within qualitative and quantitative ranges that do not impair the effects of the invention.

[0098] Examples of aqueous solutions for injection include water for injection, physiological saline, Ringer's solution, isotonic solutions containing glucose and other adjuvants, and the like. Examples of oily liquids for injection include sesame oil and soybean oil.

[0099] Stabilizers include, for example, polyethylene glycol; antioxidants and chelating agents such as sodium bisulfite, sodium metabisulfite, and ascorbic acid; and filler gases such as nitrogen and carbon dioxide.

[0100] Examples of solubilizing agents include alcohols such as ethanol; polyalcohols such as propylene glycol and polyethylene glycol; nonionic surfactants such as polysorbate 80HCO-50; benzyl benzoate; benzyl alcohol; meglumine; ethylenediamine; and nicotinamide.

[0101] Examples of buffering agents include citrates, acetates, and phosphates. Examples of soothing agents include procaine hydrochloride, lidocaine hydrochloride, benzalkonium chloride, chlorobutanol, and benzyl alcohol.

[0102] Examples of suspending agents and emulsifying agents include water-soluble polymeric substances such as carmellose sodium and sodium alginate, aluminum monostearate, fat emulsions, and lecithin.

[0103] Examples of preservatives include parahydroxybenzoic acid esters, phenol, cresol, chlorobutanol, and benzyl alcohol.

[0104] The dosage of the pharmaceutical of the present invention varies depending on the subject, administration route, target disease, symptoms, etc., but for example, when administered orally to an adult patient, the single dose is usually 0.1 to 1000 mg, preferably 1 to 200 mg, of the active ingredient, and this amount is desirably administered 1 to 3 times a day, preferably before or after meals.When administered parenterally, the single dose is usually 0.01 to 100 mg, preferably 0.1 to 20 mg, of the active ingredient, and this amount is preferably administered 1 to 3 times a day.

[0105] The pharmaceutical agent of the present invention can also be administered over a period of time. An example of the preparation of a formulation containing the amorphous compound [I] obtained by the present invention is shown below.

[0106] Formulation Example 1 Granules containing the following ingredients are prepared: (Prescription) Ingredients Compound [I] 10mg Lactose 700mg Cornstarch 274mg HPC-L 16mg 1000mg (Manufacturing method) Compound [I] and lactose are passed through a sieve. Cornstarch is passed through a sieve. These are mixed in a mixer. An aqueous solution of HPC-L is added to the mixed powder, kneaded, granulated (extrusion granulation), and then dried. The resulting dried granules are passed through a vibrating sieve to obtain granules.

[0107] Formulation Example 2 A capsule filling powder containing the following ingredients is prepared: Ingredients Compound [I] 10mg Lactose 79mg Cornstarch 10mg Magnesium stearate 1mg 100mg (Manufacturing method) Compound [I] and lactose are passed through a sieve. Cornstarch is passed through a sieve. These and magnesium stearate are mixed in a mixer to obtain a powder. The obtained powder can be filled into capsules.

[0108] Formulation Example 3 Granules for capsule filling are prepared containing the following ingredients: Ingredients Compound [I] 15mg Lactose 90mg Cornstarch 42mg HPC-L 3mg 150mg (Manufacturing method) Compound [I] and lactose are passed through a sieve. Cornstarch is passed through a sieve. These are mixed in a mixer. An aqueous solution of HPC-L is added to the mixed powder, kneaded, granulated, and then dried. The resulting dried granules are passed through a vibrating sieve and sized to obtain granules. The resulting granules can be filled into capsules.

[0109] Formulation Example 4 Tablets containing the following ingredients are prepared: Ingredients Compound [I] 10mg Lactose 90mg Microcrystalline cellulose 30mg Magnesium stearate 5mg CMC-Na 15mg 150mg (Manufacturing method) Compound [I], lactose, microcrystalline cellulose, and CMC-Na are sieved and mixed. Magnesium stearate is added to the mixed powder to obtain a mixed powder for formulation. This mixed powder is directly compressed to obtain tablets.

[0110] Formulation Example 5 An injection containing the following ingredients is prepared. In 1 ampoule (5 mL) Ingredients Compound [I] 5mg Glucose 30mg Water for injection 5mL [Example]

[0111] Next, the present invention will be explained in more detail by the following examples and test examples, but the present invention is not limited to these examples and may be modified within the scope of the present invention.

[0112] The abbreviations used in this specification have the following meanings: DMF: N,N-dimethylformamide Powder X-ray diffraction measurements were performed using SmartLab (Rigaku Corporation).

[0113] Thermogravimetry / differential thermal analysis (TG / DTA) was measured using a Thermo Plus Evo TG8120 (Rigaku Corporation).

[0114] 1 H nuclear magnetic resonance analysis (NMR) (400 MHz) was measured using an AVANCE III HD 400 (BRUKER).

[0115] Compound names were determined using ACD / Name (ACD / Name2020.1.2, Advanced Chemistry Development, Inc.). According to ChemDraw (ChemDraw Professional version 19.1, PerkinElmer Informatics, Inc.), the compound name of compound [I] is "(S)-4-((S)-2-((2S,4S)-4-amino-1-((R)-4-((4-(4-carbamoylbenzamido)phenyl)sulfonamido)-4-carboxybutanoyl)pyrrolidine-2-carboxamido)-N,4-dimethylpentanamido)-5-((5-amino-5-oxopentyl)amino)-5-oxopentanoic acid".

[0116] The CAS number of compound [I] is "2642181-22-2." For asymmetric carbons in the compounds described herein, the stereochemistry depicted indicates the absolute configuration.

[0117] A compound in which the absolute configuration of an asymmetric carbon is indicated is an optically active compound. In this specification, "room temperature" refers to 20 to 30°C unless otherwise specified.

[0118] Unless otherwise specified, "ice-cold temperature" refers to 0 to 5°C. Furthermore, "ice-cold temperature" with added salt (sodium chloride) refers to -20 to 0°C, unless otherwise specified.

[0119] The "salt" of compound [I] in this specification is not particularly limited as long as it is a pharmaceutically acceptable salt, and examples thereof include mineral acid salts such as hydrochloride, hydrobromide, hydroiodide, phosphate, sulfate, and nitrate; sulfonates such as methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and trifluoromethanesulfonate; salts with organic acids such as oxalate, tartrate, citrate, maleate, succinate, acetate, benzoate, mandelate, ascorbate, lactate, gluconate, malate, fumarate, and monosebacate; amino acid salts such as glycine salt, lysine salt, arginine salt, ornithine salt, glutamate, and aspartate; inorganic salts such as lithium salt, sodium salt, potassium salt, calcium salt, and magnesium salt; and salts with organic bases such as ammonium salt, triethylamine salt, diisopropylamine salt, and cyclohexylamine salt. The salts also include hydrated salts. The amorphous compound [I] can be produced by a known method, for example, by the method described in WO2021 / 090959.

[0120] (Example 1-1) N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N 2 Crystal of α-methyl-L-α-glutamine DMF solvate

[0121] [ka]

[0122] Amorphous N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N 2 To a solution of 1.0 g of α-methyl-L-α-glutamine trifluoroacetate in 10 mL of DMF, N,N-diisopropylethylamine (0.38 mL) and acetonitrile (5 mL) were added. After confirming complete dissolution, seed crystals (1 mg) were added and the mixture was stirred at room temperature for 24 hours. The precipitated solid was collected by filtration using a Kiriyama funnel and washed with a 2:1 DMF-acetonitrile mixture (15 mL) and acetonitrile (5 mL). The solid was dried under reduced pressure at 40°C for 4 hours to obtain a colorless solid (0.969 g).

[0123] The powder X-ray diffraction pattern of the above solid, 1 H nuclear magnetic resonance spectroscopy (NMR), infrared absorption spectroscopy, and thermogravimetry / differential thermal analysis (TG / DTA) were measured.

[0124] As shown in FIG. 1, the powder X-ray diffraction pattern showed peaks at 2θ=4.0 degrees, 8.0 degrees, 15.6 degrees, 17.8 degrees, and 21.6 degrees. In the differential thermal analysis (DTA), an endothermic peak was observed in the range of 150 to 180°C, as shown in Figure 2. In addition, a broad endothermic peak was observed in the range of 100 to 150°C.

[0125] Furthermore, thermogravimetry (TG) revealed weight loss in the ranges of 100 to 150°C and 150 to 180°C. The infrared absorption spectrum (KBr method) is shown in Figure 3, with a peak at 3358 cm -1 , 1741cm -1 , 1683cm -1 , 1527cm -1 , 1172cm -1 , and 1010 cm -1 A characteristic absorption band was observed.

[0126] 1In the H-NMR, a singlet peak was observed at 2.73 ppm, as shown in Figure 4. This peak was thought to be derived from the methyl protons of DMF. For reference, the amorphous form of compound [I] 1 The 1 H nuclear magnetic resonance analysis (NMR) is shown in Figure 5. No peak attributable to DMF was observed near 2.73 ppm.

[0127] The solvent used in the crystallization 1 Based on the characteristic peaks in the 1 H NMR chart and the information from TG / DTA, the crystals obtained above were identified as a DMF solvate. Although crystallization was attempted in the same manner as above using many solvents other than DMF, only DMF gave crystals.

[0128] Example 2-1 N-[4-(4-carbamoylbenzamido)benzene-1-sulfonyl]-D-γ-glutamyl-(4S)-4-amino-L-prolyl-L-leucyl-N-(5-amino-5-oxopentyl)-N 2 -Methyl-L-α-glutamine amorphous The crystals (150 mg) obtained in Example 1-1 were added in 10 portions to a mixed solution of ethanol (240 μL) and water (100 μL) with stirring to dissolve. This solution was added dropwise to ethanol (3.0 mL) with stirring. The residue was washed with a mixed solution of ethanol (75 μL) and water (25 μL) and added. Upon addition, a solid dispersed and precipitated. The precipitated solid was collected by filtration using a Kiriyama funnel under a nitrogen atmosphere. The obtained solid was dried under reduced pressure at room temperature for 2 days to obtain a colorless amorphous substance (97 mg).

[0129] The powder X-ray diffraction pattern of the amorphous compound [I] is shown in FIG. The MMP2 inhibitory activity of compound [I] was evaluated by the method shown in Test Example 1 below.

[0130] (Test Example 1) Inhibitory effect of compound [I] on human MMP2 The inhibitory activity of compound [I] on human MMP2 was measured by an enzyme assay using MOCAc-Pro-Leu-Gly-Leu-A2pr(Dnp)-Ala-Arg-NH2 as the substrate. 100 μg / mL of recombinant human MMP2 enzyme and 1 mmol / L 4-aminophenylmercuric acetate were mixed in a reaction solution [50 mmol / L Tris-HCl (pH 7.5), 150 mmol / L NaCl, 10 mmol / L CaCl2, 0.05% Brij L23] and incubated at 37°C for 60 minutes. The activated human MMP2 was then poured into a 96-well microplate at a final concentration of 0.7 or 7 ng / mL. Compound [I] was then diluted to various concentrations and incubated at room temperature for 15 minutes. Next, MOCAc-Pro-Leu-Gly-Leu-A2pr(Dnp)-Ala-Arg-NH2 was added to a final concentration of 5 or 16 μmol / L to initiate the enzyme reaction. After 2 hours of reaction at room temperature, the fluorescence intensity (Ex 320 nm / Em 400 nm) was measured using a microplate reader. The enzyme inhibition rate (%) was calculated using the measured fluorescence value according to the following formula, and the 50% inhibitory concentration (IC) of compound [I] was determined. 50 value) was calculated.

[0131] Enzyme inhibition rate (%) = [1-(AB) / (CB)]*100 A: Fluorescence value when compound is added B: Fluorescence value without adding compound or enzyme C: Fluorescence value without compound addition

[0132] [Table 1]

[0133] In addition, the inhibitory activity of compound [I] against various MMP subtypes can be evaluated, for example, by the method described in WO2021 / 090959.

[0134] Furthermore, the usefulness of compound [I] in improving pulmonary inflammation and fibrosis can be evaluated, for example, by the method described in WO2023 / 204170. [Industrial Applicability]

[0135] The provision of a crystalline DMF solvate of compound [I] of the present invention has made it possible to easily produce highly pure compound [I] having excellent MMP2 inhibitory activity. The present invention is expected to contribute to the development of the pharmaceutical industry by providing a pharmaceutical effective for the prevention or treatment of cancer disease or organ fibrosis, symptoms associated with cancer disease or organ fibrosis, or lung inflammation and fibrosis.

Claims

1. A crystal of a DMF solvate of the compound represented by the following formula [I]: 【Chemistry 1】

2. The crystal according to claim 1, having the following physical properties (a) to (b): (a) In powder X-ray diffraction (Cu-Kα), it has peaks at 2θ=4.0 degrees, 8.0 degrees, 15.6 degrees, 17.8 degrees, and 21.6 degrees; and (b) It has an endothermic peak at 150 to 180°C in differential thermal analysis (DTA).

3. The following formula [I]: 【Chemistry 2】 and crystallizing a DMF solvate of the compound represented by formula [I] from the mixture obtained in the step.

4. The following formula [I]: 【Transformation 3】 a solvent; and removing the solvent.

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