Process for producing uracil derivatives

Abstract

To provide a method for producing a novel uracil derivative. Formula (I): TIFF0007692546000071.tif2542 (In the formula, R 1 is halogen or the like, and Z is CH or the like.) A compound represented by the formula or a salt thereof, and formula (II): TIFF0007692546000072.tif1547 (In the formula, R 2 are each independently halogen or the like, and n is an integer of 1 to 5.) A compound represented by the formula or a salt thereof is reacted in the presence or absence of a base and in the presence of a condensing agent selected from the group consisting of T3P (registered trademark) or the like, and a method for producing a compound represented by formula (III): TIFF0007692546000073.tif3063 (The symbols in the formula have the same meanings as described above.) A method for producing a compound represented by the formula or a salt thereof.

Description

Technical Field

[0001] The present invention relates to a novel compound showing coronavirus 3CL protease inhibitory activity, or a novel synthetic intermediate thereof, or salts thereof, and a method for producing the same.

Background Art

[0002] Coronaviruses belonging to the subfamily Orthocoronavirinae of the family Coronaviridae, order Nidovirales, have a genome size of about 30 kilobases and are the largest single-stranded + strand RNA viruses among known RNA viruses. Coronaviruses are classified into four genera: Alphacoronavirus, Betacoronavirus, Gammacoronavirus, and Deltacoronavirus. As coronaviruses that infect humans, two types in the genus Alphacoronavirus (HCoV-229E, HCoV-NL63) and five types in the genus Betacoronavirus (HCoV-HKU1, HCoV-OC43, SARS-CoV, MERS-CoV, SARS-CoV-2) are known. Among these, four types (HCoV-229E, HCoV-NL63, HCoV-HKU1, HCoV-OC43) are pathogens of the common cold, while the remaining three types are severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV), Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV), and novel coronavirus (SARS-CoV-2), which cause severe pneumonia.

[0003] The novel coronavirus disease (COVID-19) that emerged in December 2019 rapidly spread across the international community and was declared a pandemic by the WHO on March 11, 2020. Droplet infection, contact infection, and aerosol infection have been reported as the main routes of SARS-CoV-2 infection, and it has been confirmed that SARS-CoV-2 can remain suspended in the air with aerosols for about 3 hours and maintain its infectivity (Non-Patent Document 1). The incubation period is about 2 to 14 days, and common cold-like symptoms such as fever (87.9%), dry cough (67.7%), fatigue (38.1%), and sputum (33.4%) are typical (Non-Patent Document 2). In severe cases, respiratory failure due to acute respiratory distress syndrome, acute lung injury, interstitial pneumonia, etc. occurs. In addition, multiple organ failure such as renal failure and liver failure has also been reported.

[0004] In Japan, from the drug repositioning of existing drugs, remdesivir, an antiviral drug, dexamethasone, an anti-inflammatory drug, and baricitinib, a rheumatism drug, have been approved as therapeutic drugs for COVID-19, and tocilizumab, an anti-IL-6 receptor antibody, was additionally approved in January 2022. In addition, in July 2021, ronapreve (casirivimab / imdevimab), an antibody cocktail therapy, was approved under special circumstances, sotrovimab was approved under special circumstances in September 2021, and molnupiravir was approved under special circumstances in December 2021. Sufficient evidence has not been obtained regarding the effectiveness and safety of these drugs. Therefore, the development of therapeutic drugs for COVID-19 is an urgent task.

[0005] When the coronavirus infects a cell, it synthesizes two polyproteins. These two polyproteins contain a replication complex that makes the viral genome and two proteases. Proteases cleave the polyproteins synthesized from the virus and play an essential role in enabling each protein to function. Among the two proteases, 3CL protease (main protease) is responsible for most of the cleavage of the polyprotein (Non-Patent Document 3). As a COVID-19 therapeutic targeting 3CL protease, in June 2021, the completion of the Phase 1b trial of Lufotrelvir (PF-07304814), a prodrug of PF-00835231 by Pfizer, was posted on ClinicalTrials.gov (NCT04535167). Also in March 2021, Pfizer announced the initiation of a Phase 1 trial of PF-07321332, a therapeutic for coronavirus disease 2019. The structural formulas of PF-00835231, Lufotrelvir, and PF-07321332 are as shown below, and they have different chemical structures from the compounds produced by the manufacturing method according to the present invention (Non-Patent Documents 4, 8, and 9, and Patent Documents 1 and 2). PF-00835231:

Chem.

Chem.

Chem.

[0006] Also, as a COVID-19 therapeutic targeting 3CL protease, in August 2021, the initiation of the Phase 1 trial of PBI-0451 by Pardes Biosciences was posted on ClinicalTrials.gov (NCT05011812). The structural formula of PBI-0451 is as shown below, and it has a different chemical structure from the compounds produced by the manufacturing method according to the present invention (Non-Patent Document 11).

Chem.

[0007] Furthermore, as a COVID-19 therapeutic targeting 3CL protease, Zocova (registered trademark) was emergently approved in Japan on November 22, 2022 (Non-Patent Document 16). The active ingredient of Zocova is ensitrelvir fumarate, and its structural formula is as shown below, and it has a different chemical structure from the compound produced by the production method according to the present invention (Patent Documents 10 and 11).

Chemical formula

[0008] On the other hand, sufficient evidence has not been obtained regarding resistance mutations to COVID-19 therapeutics targeting 3CL protease.

[0009] Compounds having 3CL protease inhibitory activity are disclosed in Non-Patent Documents 4 to 7 and 12 to 15, but neither the compound produced by the production method according to the present invention is described nor suggested in any of these documents. Also, the compound according to the present invention is described in Patent Documents 12 to 13, but the production method of the present invention is neither described nor suggested. Ρ2X 3 and / or Ρ2X 2 / 3 Compounds having P2X receptor inhibitory action are disclosed in Patent Documents 3 to 9, but neither the 3CL protease inhibitory activity nor the antiviral effect is described or suggested in any of these documents. Compounds having HIV-1 reverse transcriptase inhibitory action are described in Non-Patent Document 10, but neither the 3CL protease inhibitory activity nor the anti-coronavirus effect is described or suggested.

Prior Art Documents

Patent Documents

[0010]

Patent Document 1

Patent Document 2

[0011] [Non-Patent Document 1] The NEW ENGLAND JOURNAL of MEDICINE (2020), Vol. 382, pp. 1564 - 1567 [Non-Patent Document 2] "Report of the WHO-China Joint Mission on Coronavirus Disease 2019 (COVID-19)", [online], February 28, 2020, WHO, [searched on March 16, 2023], Internet <URL:https: / / www.who.int / docs / default-source / coronaviruse / who-china-joint-mission-on-covid-19-final-report.pdf> [Non-Patent Document 3] Science (2003), vol. 300, pp. 1763 - 1767 [Non-Patent Document 4] "A comparative analysis of SARS-CoV-2 antivirals characterizes 3CLpro inhibitor PF-00835231 as a potential new treatment for COVID-19", Journal of Virology, 2021 Mar 10;95(7), e01819 - 20 [Non-Patent Document 5] Cell Research (2020), vol. 30, pp. 678 - 692 [Non-Patent Document 6] Science (2020), vol. 368, pp. 409 - 412 [Non-Patent Document 7] ACS Central Science (2021), vol. 7, no. 3, pp. 467 - 475 [Non-Patent Document 8] 261st Am Chem Soc (ACS) Natl Meet · 2021 - 04 - 05 / 2021 - 04 - 16 · Virtual, N / A · Abst 243 [Non-Patent Document 9] Science (2021), vol. 374, pp. 1586 - 1593 [Non-Patent Document 10] Synthetic Communications (2006), vol. 36, no. 19, pp. 2913 - 2920

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 13

Non-Patent Document 14

Non-Patent Document 15

Non-Patent Document 16

Summary of the Invention

Problems to be Solved by the Invention

[0012] An object of the present invention is to provide a novel compound having coronavirus 3CL protease inhibitory activity, or a novel synthetic intermediate thereof, or a salt thereof, and a method for producing the same.

Means for Solving the Problems

[0013] The present invention relates to the following. (1) Formula (I):

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[0014] The compound represented by formula (IX) produced by the production method according to the present invention has inhibitory activity against coronavirus 3CL protease and is useful as a therapeutic and / or prophylactic agent for coronavirus infections. In addition, the compound represented by formula (IX) produced by the production method according to the present invention is useful as a pharmaceutical ingredient. Furthermore, a pharmaceutical composition containing the compound represented by formula (IX) produced by the production method according to the present invention is very useful as a therapeutic and / or prophylactic agent for coronavirus disease 2019 (COVID-19). The production method according to the present invention is a method capable of producing the compound represented by formula (IX) in a good yield. [Brief Description of the Drawings]

[0015]

Figure 1

Figure 2

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Figure 9

Figure 10

Mode for Carrying Out the Invention

[0016] The meaning of each term used in this specification will be described below. Unless otherwise specified, each term is used with the same meaning whether used alone or in combination with other terms. The term "consisting of" means having only the constituent elements. The terms "comprising" and "including" mean not being limited to the constituent elements and not excluding elements not described. Hereinafter, the present invention will be described while showing embodiments. Throughout this specification, it should be understood that the singular forms also include the plural concept thereof unless otherwise specified. Therefore, the singular articles (e.g., "a", "an", "the" in English, etc.) should be understood to include the plural concept thereof unless otherwise specified. Also, the terms used in this specification should be understood to be used in the ordinary meaning usually used in the above field unless otherwise specified. Therefore, unless otherwise defined, all technical terms and scientific and technical terms used in this specification have the same meaning as generally understood by those skilled in the art to which the present invention pertains. In case of contradiction, this specification (including the definitions) shall prevail.

[0017] "Halogen" includes a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. Particularly, a fluorine atom and a chlorine atom are preferred.

[0018] "Alkyl" includes a linear or branched hydrocarbon group having 1 to 15 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, and still more preferably 1 to 4 carbon atoms. For example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-heptyl, isoheptyl, n-octyl, isooctyl, n-nonyl, n-decyl, etc. can be mentioned. Preferred embodiments of "alkyl" include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and n-pentyl. More preferred embodiments include methyl, ethyl, n-propyl, isopropyl, and tert-butyl. "C1-C4 alkyl" includes a linear or branched hydrocarbon group having 1 to 4 carbon atoms. For example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, etc. can be mentioned. Preferred embodiments of "C1-C4 alkyl" include methyl, ethyl, n-propyl, and isopropyl. "Halo C1-C4 alkyl" means a group in which one or more of the above "halogens" are bonded to the above "C1-C4 alkyl". When substituted with two or more halogens, the halogens may be the same or different. For example, monofluoromethyl, difluoromethyl, 2-monofluoroethyl, 3-monofluoropropyl, 3,3,3-trifluoropropyl, 2,2,3,3,3-pentafluoropropyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, 2,2-difluoroethyl, 1,1,1-trifluoropropan-2-yl, 4,4,4-trifluorobutyl, 3,3,4,4,4-pentafluorobutyl, etc. may be mentioned. Preferred embodiments of "halo C1-C4 alkyl" include trifluoromethyl, 2,2,2-trifluoroethyl, and 3,3,3-trifluoropropyl.

[0019] The compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) are not limited to specific isomers, but include all possible isomers (for example, keto-enol isomers, imine-enamine isomers, diastereoisomers, optical isomers, rotational isomers, etc.), racemates or mixtures thereof. For example, the compound represented by formula (V) includes the following tautomers.

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[0020] One or more hydrogens, carbons, and / or other atoms of the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) may each be substituted with isotopes of hydrogen, carbon, and / or other atoms. Examples of such isotopes include, respectively 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, 123 I and 36 Cl, including hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine and chlorine. The compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) also include compounds substituted with such isotopes. The compounds substituted with such isotopes are also useful as pharmaceuticals. The compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) include all radiolabels substituted with radioisotopes contained in such isotopes. Also included in the present invention is a "radiolabeling method" for producing the "radiolabel", and the "radiolabel" is useful as a tool for metabolic pharmacokinetic studies, studies in binding assays and / or diagnosis.

[0021] Radioactive labels of the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) can be prepared by methods well-known in the art. For example, tritium-labeled compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) can be prepared by introducing tritium into specific compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) by a catalytic dehalogenation reaction using tritium. This method involves reacting a suitably halogen-substituted precursor of the compound represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) with tritium gas in the presence or absence of a base, in the presence of a suitable catalyst such as Pd / C. Other suitable methods for preparing tritium-labeled compounds can be referred to "Isotopes in the Physical and Biomedical Sciences, Vol. 1, Labeled Compounds (Part A), Chapter 6 (1987)". 14 C-labeled compounds can be 14 prepared by using starting materials having C carbon.

[0022] The compounds represented by formula (IX) used in the present specification may form prodrugs. A prodrug is a derivative of a compound produced by the production method according to the present invention having a group that can be chemically or metabolically decomposed, and is a pharmaceutically active compound by solvolysis or in vivo under physiological conditions. Prodrugs include compounds that are enzymatically oxidized, reduced, hydrolyzed, etc. under physiological conditions in vivo to be converted into the compound represented by formula (IX), compounds that are hydrolyzed by gastric acid, etc. to be converted into the compound represented by formula (IX), and the like. Methods for selecting and producing appropriate prodrug derivatives are described, for example, in "Design of Prodrugs, Elsevier, Amsterdam, 1985". Prodrugs may themselves have activity.

[0023] The "compound represented by formula (IX)" used in the present specification may form salts, co-crystals, or solvates thereof. The "compound represented by formula (IX), or a salt thereof" used in the present specification also includes such various salts, co-crystals, and solvates thereof.

[0024] As used herein, the "salt" means, for example, that the counter molecule is regularly arranged within the same crystal lattice as the compound represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX), and formula (X), and may contain any number of counter molecules. It refers to those via ionic bonds by proton transfer between the compound and the counter molecule in the crystal lattice.

[0025] Examples of salts of the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) include salts of the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) with alkali metals (such as lithium, sodium, potassium, etc.), alkaline earth metals (such as calcium, barium, etc.), magnesium, transition metals (such as zinc, iron, etc.), ammonia, organic bases (such as trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine, triethanolamine, meglumine, ethylenediamine, pyridine, picoline, quinoline, etc.) and amino acids, or inorganic acids (such as hydrochloric acid, sulfuric acid, nitric acid, carbonic acid, hydrobromic acid, phosphoric acid, hydroiodic acid, etc.), and organic acids (such as formic acid, acetic acid, propionic acid, trifluoroacetic acid, citric acid, lactic acid, tartaric acid, oxalic acid, maleic acid, fumaric acid, succinic acid, mandelic acid, glutaric acid, malic acid, benzoic acid, phthalic acid, ascorbic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, trifluoroacetic acid, etc.). These salts can be formed by conventional methods.

[0026] The compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) of the present invention or salts thereof may form solvates (e.g., hydrates, etc.), co-crystals and / or crystal polymorphs, and the present invention also encompasses such various solvates, co-crystals and crystal polymorphs. A "solvate" may be coordinated with any number of solvent molecules (e.g., water molecules, etc.) with respect to the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X). When the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) or salts thereof or their solvates are left in the air, they may absorb moisture and adsorbed water may adhere, or hydrates may be formed. Also, crystal polymorphs may be formed by recrystallizing the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) or salts thereof or their solvates. A "co-crystal" means that the compound represented by formula (I) or a salt and a co-former molecule are present in the same crystal lattice and may contain any number of co-former molecules.

[0027] As used herein, "co-crystal" means that co-former molecules are regularly arranged in the same crystal lattice and may contain any number of co-former molecules. Also, a co-crystal refers to a case where the intermolecular interaction between a compound and a co-former molecule is mediated by non-covalent and non-ionic chemical interactions such as hydrogen bonds and van der Waals forces.

[0028] Generally, it is considered that proton transfer occurs between a compound and a counter molecule in a salt, but it is also known that in some cases, the proton transfer may not be complete. Since this state is not a true salt, it may be called a co-crystal. It is also known that proton transfer may vary continuously with temperature. Therefore, the "salt of the compound represented by formula (IX)" as used herein includes co-crystals and refers to the salt or co-crystal of the compound represented by formula (IX).

[0029] One aspect herein is a pharmaceutically acceptable salt or co-crystal of the compound represented by formula (IX) with hydrofluoric acid, hydrochloric acid, hydrobromic acid, orthophosphoric acid, hydroiodic acid, nitric acid, phosphoric acid, boric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, trifluoromethylbenzenesulfonic acid, chlorobenzenesulfonic acid, methoxybenzenesulfonic acid, acetic acid, propionic acid, lactic acid, citric acid, fumaric acid, malonic acid, malic acid, succinic acid, salicylic acid, maleic acid, glycerophosphoric acid, tartaric acid, benzoic acid, glutamic acid, aspartic acid, 2-naphthalenesulfonic acid, hexanoic acid, acetylsalicylic acid, etc.

[0030] The study of salt formation and co-crystal formation provides a means to change the physicochemical characteristics and the resulting biological characteristics of a drug without changing its chemical structure. Salt formation and co-crystal formation can have a dramatic impact on the properties of a drug. In the selection of an appropriate salt or co-crystal, hygroscopicity, stability, solubility, and processing characteristics are also important aspects. The solubility of a salt or co-crystal can affect its suitability for use as a drug. If the water solubility is low, the dissolution rate in in vivo administration is rate-limiting in the absorption process, which may result in low bioavailability. Also, due to low water solubility, administration by injection may become difficult, which may limit the choice of an appropriate administration route.

[0031] The complex containing the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) of the present invention broadly includes salts, co-crystals and inclusion compounds, or solvates thereof.

[0032] The "compound represented by formula (IX)" can form a solvate with water (i.e., hydrate) or a solvate with a general organic solvent. The "salt of the compound represented by formula (IX)" can form a solvate with water (i.e., hydrate) or a solvate with a general organic solvent.

[0033] As used herein, "solvate" refers to, for example, a compound represented by formula (I), formula (II), formula (III), formula (III-1), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X), which is regularly arranged with any number of solvent molecules. Examples of the solvent molecule include ethyl acetate, water, ethanol, acetone, 1,1 - diethoxypropane, 1,1 - dimethoxymethane, 2,2 - dimethoxypropane, isooctane, isopropyl ether, methyl isopropyl ketone, methyl tetrahydrofuran, petroleum ether, trichloroacetic acid, trifluoroacetic acid, acetic acid, anisole, 1 - butanol, 2 - butanol, n - butyl acetate, t - butyl methyl ether, cumene, dimethyl sulfoxide, diethyl ether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3 - methyl - 1 - butanol, methyl ethyl ketone, methyl isobutyl ketone, 2 - methyl - 1 - propanol, pentane, 1 - pentanol, 1 - propanol, 2 - propanol, propyl acetate, tetrahydrofuran, acetonitrile, chlorobenzene, chloroform, cyclohexane, 1,2 - dichloroethene, dichloromethane, 1,2 - dimethoxyethane, N,N - dimethylacetamide, N,N - dimethylformamide, 1,4 - dioxane, 2 - ethoxyethanol, ethylene glycol, formamide, hexane, methanol, 2 - methoxyethanol, methyl butyl ketone, methyl cyclohexane, N - methylpyrrolidone, nitromethane, pyridine, sulfolane, tetralin, toluene, 1,1,2 - trichloroethene, xylene, and t - butanol. Preferably, ethyl acetate, water, ethanol, acetone, 1,1 - diethoxypropane, 1,1 - dimethoxymethane, 2,2 - dimethoxypropane, isooctane, isopropyl ether, methyl isopropyl ketone, methyl tetrahydrofuran, petroleum ether, trichloroacetic acid, trifluoroacetic acid, acetic acid, anisole, 1 - butanol, 2 - butanol, n - butyl acetate, t - butyl methyl ether, cumene, dimethyl sulfoxide, diethyl ether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3 - methyl - 1 - butanol, methyl ethyl ketone, methyl isobutyl ketone, 2 - methyl - 1 - propanol, pentane, 1 - pentanol, 1 - propanol, 2 - propanol, propyl acetate, toluene, and tetrahydrofuran are included. More preferably, ethyl acetate, water, ethanol, acetone, 1,1 - diethoxypropane, 1,1 - dimethoxymethane, 2,2 - dimethoxypropane, isooctane, isopropyl ether, methyl isopropyl ketone, methyl tetrahydrofuran, petroleum ether, toluene, trichloroacetic acid, trifluoroacetic acid, etc. may be mentioned. As the solvate of the compound represented by the formula (VI), a toluene solvate is preferable. In addition, when the compounds represented by the formula (I), formula (II), formula (III), formula (III - 1), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) are left in the air, they may absorb moisture and adsorbed water may adhere, or a hydrate may be formed.

[0034] In addition, the "crystal" used in this specification means a solid in which atoms, ions, molecules, etc. that make up it are arranged regularly three - dimensionally, and is distinguished from an amorphous solid that does not have such a regular internal structure. The crystals used in this specification may be single crystals, twins, polycrystals, etc. Furthermore, there may be "crystal polymorphs" in which the composition is the same but the arrangement in the crystal is different in the "crystal", and these are collectively referred to as "crystal forms". The "compound represented by the formula (IX), or a salt thereof" includes their crystal polymorphs. The crystals used in this specification may be deuterium - converted forms. The crystals used in this specification may be labeled with isotopes (e.g., 3 H, 14 C, 35 S, 125 I, etc.). The crystal form and / or crystallinity can be confirmed by spectroscopic methods such as X - ray diffraction, Raman spectroscopy, infrared absorption spectroscopy, solid - state NMR, etc. Also, the physical properties of the crystal can be confirmed by many techniques such as differential scanning calorimetry, moisture adsorption - desorption measurement, dissolution properties, etc.

[0035] One aspect in this specification is an anhydrous crystal of the compound represented by the formula (IX). As used herein, "anhydride" is synonymous with "non-solvate", "unsolvated", "anhydrate" and "non-hydrate". The anhydrous crystal of the compound represented by formula (IX) has a theoretical water content of 0% by weight. However, in the analysis of the water content and / or solvent content, it may take a value higher than the theoretical water content of the crystal due to the influence of the adsorbed water and / or adsorbed solvent adhering to the crystal surface.

[0036] One embodiment herein is an anhydrous crystal of the compound represented by formula (IX) having characteristic peaks at diffraction angles (2θ): 6.5° ± 0.2°, 15.6° ± 0.2°, 17.4° ± 0.2°, 19.9° ± 0.2° and 20.3° ± 0.2° in a powder X-ray diffraction pattern (CuKα ray, λ = 1.5418 Å).

[0037] One embodiment herein is when a single crystal diffraction experiment is carried out at 298 K (25 °C) with a CuKα ray (λ = 1.5418 Å). The following crystallographic data: Space group: Pbca a = 14.67 Å ± 0.05 Å b = 11.83 Å ± 0.05 Å c = 27.10 Å ± 0.05 Å α = 90° β = 90° γ = 90° An anhydrous crystal of the compound represented by formula (IX) characterized by the above.

[0038] One embodiment herein is an anhydrous crystal of the compound represented by formula (IX) having a melting point of 261.3 °C ± 2 °C in differential scanning calorimetry (DSC).

[0039] One embodiment herein is an anhydrous crystal of the compound represented by formula (IX) having a melting point of 265.6 °C ± 2 °C in simultaneous differential thermal-thermogravimetric measurement (TG / DTA).

[0040] One embodiment herein is in the Raman spectrum, 415.2 cm -1 ± 2 cm-1 、 502.7 cm -1 ± 2 cm -1 、 1431.4 cm -1 ± 2 cm -1 、 1714.8 cm -1 ± 2 cm -1 and 3065.4 cm -1 ± 2 cm -1 and having characteristic peaks at ± 2 cm, is an anhydrous crystal of the compound represented by formula (IX).

[0041] (Powder X-ray diffraction (XRPD)) Powder X-ray diffraction (XRPD) is one of the most sensitive analytical methods for measuring the crystalline form and crystallinity of solids. When X-rays irradiate a crystal, they are reflected by the crystal lattice planes and interfere with each other, showing diffraction lines corresponding to the periodicity of the structure. On the other hand, for an amorphous solid, since it usually does not have an ordered repeating period in its structure, the diffraction phenomenon does not occur, and it shows a broad XRPD pattern without features (also called a halo pattern).

[0042] The crystalline forms of the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) can be identified by powder X-ray diffraction patterns and characteristic diffraction peaks. The crystalline forms of the compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX) and formula (X) can be distinguished from other crystalline forms by the presence of characteristic diffraction peaks. The characteristic diffraction peaks used in this specification are peaks selected from the observed diffraction patterns. The characteristic diffraction peaks are preferably selected from about 10 peaks, more preferably about 5 peaks, and even more preferably about 3 peaks in the diffraction pattern. In differentiating multiple crystals, peaks that are identified in a particular crystal and not in other crystals are preferable characteristic peaks for identifying that crystal, rather than peak intensities. Even one or two such characteristic peaks can characterize the crystal. If the measured charts are compared and these characteristic peaks match, it can be said that the powder X-ray diffraction spectra substantially match.

[0043] Generally, since errors can occur in the diffraction angle (2θ) in powder X-ray diffraction within a range of ±0.2°, the values of the diffraction angles in powder X-ray diffraction should be understood to include numerical values within a range of about ±0.2°. Therefore, the present invention includes not only crystals in which the diffraction angles of the peaks completely match, but also crystals in which the diffraction angles of the peaks match with an error of about ±0.2°.

[0044] The peak intensities shown in the following tables and figures generally can vary due to many factors, such as the effect of the preferred orientation of the crystal with respect to the X-ray beam, the influence of coarse particles, the purity of the substance being analyzed, or the crystallinity of the sample. Also, the peak positions can shift based on variations in the sample height. Furthermore, different shifts are obtained according to the Bragg equation (nλ = 2dsinθ) when measuring using different wavelengths, and compounds shown in different XRPD patterns obtained by using such different wavelengths are also included in the scope of the present invention.

[0045] (Single crystal structure analysis) One of the methods for identifying crystals is to obtain crystallographic parameters, atomic coordinates (values indicating the spatial positional relationship of each atom), and a three-dimensional structure model in the crystal. Refer to "Guide to X-ray Structure Analysis" written by Toshio Sakurai, published by Shoeka Publishing Co., Ltd. (1983), "X-Ray Structure Determination: A Practical Guide" written by Stout & Jensen, Macmillan Co., New York (1968), etc. Single crystal structure analysis is useful when identifying the structures of crystals of the complex, salt, optical isomer, tautomer, and geometric isomer according to the present invention.

[0046] (Raman spectroscopy) The Raman spectrum shows the characteristics of the vibrations of a molecule or a complex system. Its origin lies in the inelastic collision between the molecule and the photon, which is a particle of light including the light beam. The collision between the molecule and the photon results in an energy exchange, and as a result, the energy changes, causing the wavelength of the photon to change. That is, since the Raman spectrum is a very narrow spectral line emitted when a photon is incident on the target molecule, a laser or the like is used as the light source. The wavelength of each Raman line is indicated by the wavenumber shift from the incident light, which is the difference between the reciprocal of the wavelength of the Raman line and that of the incident light. The Raman spectrum measures the vibrational state of a molecule, which is determined by its molecular structure. Generally, the Raman spectrum peak (cm -1 ) may have an error within the range of ±2 cm -1 . Therefore, it is necessary to understand that the value of the above Raman spectrum peak also includes numerical values within the range of about ±2 cm -1 . Thus, not only crystals with completely matching Raman spectrum peaks in the Raman spectrum, but also crystals with Raman spectrum peaks matching with an error of about ±2 cm -1 are included in the present invention.

[0047] (Differential scanning calorimetry (DSC)) DSC is one of the main measurement methods of thermal analysis and is a method for measuring the thermal properties of a substance as an aggregate of atoms and molecules. By DSC, the change in heat quantity related to the temperature or time of the pharmaceutical active ingredient is measured, and a differential scanning calorimetry curve is obtained by plotting the resulting data against temperature or time. From the differential scanning calorimetry curve, information regarding the onset temperature when the pharmaceutical active ingredient melts, the maximum value of the endothermic peak curve associated with melting, and the enthalpy can be obtained. Regarding DSC, it is known that the observed temperature can depend on the rate of temperature change as well as the sample preparation technique and specific apparatus used. Therefore, the "melting point" in DSC refers to the onset temperature that is less susceptible to the influence of the sample preparation technique. The error range at the onset temperature obtained from the differential scanning calorimetry curve is approximately ±2 °C. In the identification of crystal identity, not only the melting point but also the overall pattern is important, and it can vary somewhat depending on the measurement conditions and measuring instrument.

[0048] (Differential Thermal - Thermogravimetric Simultaneous Measurement Method (TG / DTA)) TG / DTA is one of the main measurement methods in thermal analysis and is a method for measuring the weight and thermal properties of substances as aggregates of atoms and molecules. TG / DTA is a method for measuring the changes in weight and heat quantity related to the temperature or time of the pharmaceutical active ingredient, and by plotting the resulting data against temperature or time, TG (thermogravimetric) and DTA (differential thermal) curves are obtained. From the TG / DTA curve, information on the weight and heat quantity changes related to the decomposition, dehydration, oxidation, reduction, sublimation, and evaporation of the pharmaceutical active ingredient can be obtained. Regarding TG / DTA, it is known that the observed temperature and weight change can depend on the rate of temperature change as well as the sample preparation technique and specific apparatus used. Therefore, the "melting point" in TG / DTA refers to the onset temperature that is less susceptible to the influence of the sample preparation technique. In the identification of crystal identity, not only the melting point but also the overall pattern is important, and it can vary somewhat depending on the measurement conditions and measuring instrument.

[0049] The compound represented by formula (IX) produced by the production method according to the present invention has coronavirus 3CL protease inhibitory activity, and thus is useful as a therapeutic agent and / or prophylactic agent for diseases involving coronavirus 3CL protease. Examples of diseases involving coronavirus 3CL protease include viral infections, preferably coronavirus infections. In one aspect, examples of coronaviruses include human-infecting coronaviruses. Examples of human-infecting coronaviruses include HCoV-229E, HCoV-NL63, HCoV-HKU1, HCoV-OC43, SARS-CoV, MERS-CoV, and / or SARS-CoV-2. In one aspect, examples of coronaviruses include alpha coronaviruses and / or beta coronaviruses, more preferably beta coronaviruses, and even more preferably sarbecoviruses. In one aspect, examples of alpha coronaviruses include HCoV-229E and HCoV-NL63. Particularly preferably, HCoV-229E is included. In one aspect, examples of beta coronaviruses include HCoV-HKU1, HCoV-OC43, SARS-CoV, MERS-CoV, and / or SARS-CoV-2. Preferably, HCoV-OC43 or SARS-CoV-2, and particularly preferably SARS-CoV-2 is included. In one aspect, examples of beta coronaviruses include beta-coronavirus lineage A, beta-coronavirus lineage B, and beta-coronavirus lineage C. More preferably, beta-coronavirus lineage A and beta-coronavirus lineage B, and particularly preferably beta-coronavirus lineage B is included. Examples of β-coronavirus lineage A include, for example, HCoV-HKU1 and HCoV-OC43, preferably HCoV-OC43. Examples of β-coronavirus lineage B include, for example, SARS-CoV and SARS-CoV-2, preferably SARS-CoV-2. Examples of β-coronavirus lineage C preferably include MERS-CoV. In one aspect, examples of coronaviruses include HCoV-229E, HCoV-OC43, and / or SARS-CoV-2, particularly preferably SARS-CoV-2. It is generally known that viruses mutate during repeated growth and infection. The above-mentioned coronaviruses include not only mutant strains known in the art but also mutant strains that will emerge in the future, as long as the compound represented by formula (IX) produced by the production method according to the present invention can exhibit coronavirus 3CL protease inhibitory activity. Examples of known mutant strains of SARS-CoV-2 include, for example, the mutant strains used in the examples of this specification. Examples of coronavirus infections include infections caused by HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKU1, SARS-CoV, MERS-CoV, and / or SARS-CoV-2. Preferably, infections caused by HCoV-229E, HCoV-OC43, and / or SARS-CoV-2, particularly preferably infections caused by SARS-CoV-2. Examples of coronavirus infections particularly preferably include coronavirus disease 2019 (COVID-19).

[0050] The production method according to the present invention will be described below. Step A: Method for producing the compound represented by formula (III)

Chemical formula

[0051] The compound represented by formula (IX) produced by the production method according to the present invention has coronavirus 3CL protease inhibitory activity, and is therefore useful as a therapeutic and / or prophylactic agent for viral infectious diseases. Furthermore, the compound represented by formula (IX) produced by the production method according to the present invention has utility as a medicine and preferably has any one or a plurality of the following excellent characteristics. a) It has a weak inhibitory effect on CYP enzymes (for example, CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4, etc.). b) It shows good pharmacokinetics such as high bioavailability and moderate clearance. c) It has high metabolic stability. d) It does not show an irreversible inhibitory effect on CYP enzymes (for example, CYP3A4) within the concentration range of the measurement conditions described herein. e) It is not mutagenic. f) It has a low cardiovascular risk. g) It shows high solubility. h) It has a high protein unbound fraction (fu value). i) It has high selectivity for coronavirus 3CL protease. j) It has high coronavirus growth inhibitory activity. For example, it has high coronavirus growth inhibitory activity in the presence of human serum (HS) or human serum albumin (HSA). k) It also has high growth inhibitory activity against 3CL protease inhibitor-resistant viruses. Examples of the coronavirus growth inhibitor include, for example, in the CPE suppression effect confirmation test (SARS-CoV-2) described below, for example, EC 50 is 10 μM or less, preferably 1 μM or less, more preferably 100 nM or less.

[0052] The pharmaceutical composition containing the compound represented by formula (IX) produced by the production method according to the present invention can be administered by any method, either orally or parenterally. Examples of parenteral administration methods include transdermal, subcutaneous, intravenous, intraarterial, intramuscular, intraperitoneal, transmucosal, inhalation, intranasal, ophthalmic, otic, intravaginal administration, etc.

[0053] In the case of oral administration, it may be prepared into any of the commonly used dosage forms such as internal solid preparations (e.g., tablets, powders, granules, capsules, pills, films, etc.), internal liquid preparations (e.g., suspensions, emulsions, elixirs, syrups, lemonades, spirits, aromatic waters, extracts, decoctions, tinctures, etc.) according to conventional methods and then administered. Tablets may be sugar-coated tablets, film-coated tablets, enteric-coated tablets, sustained-release tablets, troches, sublingual tablets, buccal tablets, chewable tablets or orally disintegrating tablets. Powders and granules may be dry syrups. Capsules may be soft capsules, microcapsules or sustained-release capsules.

[0054] In the case of parenteral administration, it can be preferably administered in any of the commonly used dosage forms such as injections, drip infusions, external preparations (e.g., eye drops, nasal drops, ear drops, aerosols, inhalants, lotions, injections, coatings, gargles, enemas, ointments, plasters, jellies, creams, patches, poultices, external powders, suppositories, etc.). Injections may be emulsions such as O / W, W / O, O / W / O, W / O / W type, etc.

[0055] An effective amount of the compound represented by formula (IX) produced by the production method according to the present invention can be mixed with various pharmaceutical additives such as excipients, binders, disintegrants, lubricants, etc. suitable for the dosage form as needed to form a pharmaceutical composition. Furthermore, the pharmaceutical composition can be made into a pharmaceutical composition for pediatric use, elderly use, critically ill patients or surgical use by appropriately changing the effective amount of the compound represented by formula (IX) produced by the production method according to the present invention, the dosage form and / or various pharmaceutical additives. For example, a pharmaceutical composition for pediatric use can be administered to patients who are neonates (less than 4 weeks after birth), infants (4 weeks to less than 1 year after birth), toddlers (1 year or older and less than 7 years old), children (7 years or older and less than 15 years old) or 15 to 18 years old. For example, a pharmaceutical composition for elderly use can be administered to patients 65 years old or older.

[0056] The dosage of the pharmaceutical composition containing the compound represented by formula (IX) produced by the production method according to the present invention is preferably set in consideration of the patient's age, body weight, type and degree of disease, administration route, etc. When administered orally, it is usually 0.01 to 100 mg / kg / day, preferably in the range of 0.05 to 50 mg / kg / day. In the case of parenteral administration, it varies greatly depending on the administration route, but is usually 0.005 to 200 mg / kg / day, preferably in the range of 0.01 to 100 mg / kg / day. It may be administered once a day or divided into several times a day.

[0057] The compound represented by formula (IX) produced by the production method according to the present invention may be used in combination with, for example, a therapeutic agent for other novel coronavirus infections (COVID-19) (including approved drugs and drugs under development or to be developed in the future) (hereinafter referred to as a combined drug) for the purpose of enhancing the action of the compound or reducing the dosage of the compound. In this case, the administration time of the compound represented by formula (IX) produced by the production method according to the present invention and the combined drug is not limited, and they may be administered simultaneously or at different times to the administration subject. Furthermore, the compound represented by formula (IX) produced by the production method according to the present invention and the combined drug may be administered as two or more types of preparations containing their respective active ingredients, or may be administered as a single preparation containing their active ingredients.

[0058] The dosage of the combined drug can be appropriately selected based on the clinically used dosage. Also, the mixing ratio of the compound represented by formula (IX) produced by the production method according to the present invention and the combined drug can be appropriately selected depending on the administration subject, administration route, target disease, symptoms, combination, etc. For example, when the administration subject is a human, 0.01 to 100 parts by weight of the combined drug may be used per 1 part by weight of the compound represented by formula (IX) produced by the production method according to the present invention.

Examples

[0059] The present invention will be described in more detail below with reference to Examples, Reference Examples, and Test Examples, but the present invention is not limited thereto.

[0060] In addition, the abbreviations used in this specification have the following meanings. Boc: tert-Butoxycarbonyl CDI: N,N'-Carbonyldiimidazole ClP(O)(OPh) 2 : Diphenyl chlorophosphate cps: counts per seconds deg: ° DBU: 1,8-Diazabicyclo[5.4.0]-7-undecene DMA: N,N-Dimethylacetamide DME: 1,2-Dimethoxyethane DMF: N,N-Dimethylformamide DMI: 1,3-Dimethyl-2-imidazolidinone DMSO: Dimethyl sulfoxide DTT: Dithiothreitol EDC: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide EDT: 1,2-Ethanedithiol EDTA: Ethylenediaminetetraacetic acid Et 3 N: Triethylamine FBS: Fetal bovine serum H 2 O: Water HATU: O-(7-Azabenzotriazol-1-yl)-N,N,N’,N’,-tetramethyluronium hexafluorophosphate HCl: Hydrogen chloride HOBT: 1-Hydroxybenzotriazole LDA: Lithium diisopropylamide MEM: Eagle's minimum essential medium MsCl: Methanesulfonyl chloride PhOPOCl 2 : Phenyl dichlorophosphate PhPOCl2 : Dichlorophenylphosphonic acid POCl 3 : Phosphorus oxychloride PyBOP (registered trademark): (Benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate T3P (registered trademark): Propylphosphonic anhydride THF: Tetrahydrofuran n-Bu 4 NCl: Tetrabutylammonium chloride n-BuOH: n-Butanol t-BuOK: Potassium tert-butoxide TFA: Trifluoroacetic acid WSCD (also known as EDC): N-(3-Dimethylaminopropyl)-N’-ethylcarbodiimide WSCD·HCl: N-(3-Dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride mM: mmol / L μM: μmol / L nM: nmol / L

[0061] (Method for identifying compounds) The NMR analysis obtained in each example and reference example was performed at 400 MHz using DMSO-d 6 , CDCl 3 . Also, when showing NMR data, there are cases where not all measured peaks are described. In the specification, “RT” or “retention time” refers to the retention time in LC / MS: Liquid chromatography / mass spectrometry or high performance liquid chromatography (HPLC), measured under the following conditions. The unit “min” of “RT” means “minute”. (Measurement condition 1) Column: XBridge C18 (3.5 μm i.d. 4.6 x 150 mm) Column temperature: Constant temperature around 40 °C UV detection wavelength: 254 nm Mobile phase: [A] is an aqueous solution containing 0.1% formic acid, [B] is acetonitrile for liquid chromatography Gradient: Maintain 5% solvent [B] for 2 minutes, perform a linear gradient of 5% - 95% solvent [B] over 18 minutes, and then maintain 95% solvent [B] for 5 minutes. Flow rate: 1.0 mL / min Injection volume: 10 μL (Measurement condition 2) Column: ACQUITY UPLC (registered trademark) BEH C18 (1.7 μm i.d. 2.1x100 mm) (Waters) Column temperature: Constant temperature around 30 °C Flow rate: 0.4 mL / min UV detection wavelength: 247 nm Injection volume: 3 μL Mobile phase: [A] is an aqueous solution containing 0.1% formic acid, [B] is an acetonitrile solution containing 0.1% formic acid Gradient: Perform a linear gradient of 15% - 35% solvent [B] over 5 minutes, then maintain 35% solvent [B] for 20 minutes, then perform a linear gradient of 35% - 80% solvent [B] over 5 minutes, and then maintain 80% solvent [B] for 3 minutes. (Measurement condition 3) Column: ACQUITY UPLC (registered trademark) BEH C18 (1.7 μm i.d. 2.1x100 mm) (Waters) Flow rate: 0.4 mL / min UV detection wavelength: 247 nm Mobile phase: [A] is a 0.1% formic acid aqueous solution, [B] is an acetonitrile solution containing 0.05% formic acid Gradient: Perform a linear gradient of 15% - 35% solvent [B] over 10 minutes, then maintain 35% solvent [B] for 25 minutes, then perform a linear gradient of 35% - 80% solvent [B] over 5 minutes, and then maintain 80% solvent [B] for 3 minutes. (Measurement condition 4) Column: Titan C18 (1.9 μm i.d. 2.1x50 mm) Column temperature: Constant temperature around 40 °C UV detection wavelength: 254 nm Flow rate: 0.6 mL / min Injection volume: 1 μL Mobile phase: [A] is an aqueous solution containing 0.1% formic acid, [B] is acetonitrile for liquid chromatography Gradient: A linear gradient of 5% - 95% solvent [B] was carried out over 8 minutes, and then 95% solvent [B] was maintained for 0.5 minutes. (Measurement condition 5) Column: Titan C18 (1.9μm i.d. 2.1x50mm) Column temperature: A constant temperature near 40°C UV detection wavelength: 254nm Flow rate: 0.6 mL / min Injection volume: 1 μL Mobile phase: [A] is an aqueous solution containing 0.1% formic acid, [B] is acetonitrile for liquid chromatography Gradient: A linear gradient of 5% - 95% solvent [B] was carried out over 3 minutes, and then 95% solvent [B] was maintained for 0.5 minutes.

[0062] Powder X-ray diffraction experiment (XRPD) According to the powder X-ray diffraction measurement method described in the general test method of the Japanese Pharmacopoeia, powder X-ray diffraction measurement of the solid form (crystalline and amorphous) obtained in the examples was carried out. The measurement conditions are shown below. Measurement condition 1: Powder X-ray diffractometer: MinFlex600 manufactured by Rigaku Measurement method: Reflection method Wavelength used: CuKα ray Tube current: 15 mA Tube voltage: 40 kV Sample plate: Aluminum X-ray incident angle (θ): 4 - 40° Sampling width: 0.02° Measurement condition 2: Powder X-ray diffractometer: SmartLab manufactured by Rigaku Measurement method: Reflection method Wavelength used: CuKα ray (λ = 1.5418 Å) Tube current: 200 mA Tube voltage: 45 kV Sample plate: Aluminum X-ray incident angle: 2.5° Sampling width: 0.02° Detector: HyPix-3000 (two-dimensional detection mode)

[0063] Measurement and analysis method for single crystal structure analysis Single crystal structure analysis of the crystals obtained in the examples was performed. The measurement conditions and analysis methods are shown below. (Equipment) XtaLAB P200 MM007 manufactured by Rigaku (Measurement conditions) Measurement temperature: 25 °C Temperature controller: Low-temperature sample spraying device manufactured by Rigaku Wavelength used: CuKα ray (λ = 1.5418 Å) Software: CrysAlisPro 1.171.39.46e (Rigaku Oxford Diffraction, 2018) (Data processing) Software: CrysAlisPro 1.171.39.46e (Rigaku Oxford Diffraction, 2018) The data was subjected to Lorentz and polarization corrections and absorption correction. (Crystal structure analysis) Phase determination was carried out using the direct method program ShelXT (Sheldrick, G.M., 2015), and refinement was performed using ShelXL (Sheldrick, G.M., 2015) with the full-matrix least-squares method. The temperature factors of non-hydrogen atoms were all refined anisotropically. Hydrogen atoms were introduced by calculation using the default parameters of ShelXL unless otherwise specified and were treated as riding atoms. Also, hydrogen atoms were refined with isotropic parameters. For the drawing of the following structural diagrams, PLATON (Spek, 1991) / ORTEP (Johnson, 1976) was used (30% PROBABILITY level).

[0064] Measurement of Raman spectrum The measurement conditions for the measurement of the Raman spectrum of the crystals obtained in the examples and the baseline correction are shown below. Measurement condition 1 Measurement method: Microscopic laser Raman spectroscopy Laser wavelength: 671 nm Number of integrations: 1 time Exposure time: 1 second

[0065] Differential scanning calorimetry (DSC) The DSC of the crystal obtained in the example was measured. The sample was weighed into an aluminum pan, simply sealed, and measured. The measurement conditions are shown below. Note that the measurement by differential scanning calorimetry (DSC) may have an error within the range of ±2°C. Apparatus: Discovery DSC / TA Instrument Measurement temperature range: -10°C - 270°C Heating rate: 10°C / min Atmosphere: N 2 50 mL / min

[0066] Simultaneous measurement of differential thermal - thermogravimetry (TG / DTA) The simultaneous measurement of differential thermal - thermogravimetry (TG / DTA) of the solid form (crystal) obtained in the example was carried out. The sample obtained in the example was weighed, filled into an aluminum pan, and measured in an open system. The measurement conditions are as follows. Apparatus: Hitachi High - Technologies TG / DTA STA7200RV Measurement temperature range: Room temperature - 350°C Heating rate: 10°C / min

Example

[0067] Synthesis of the anhydrous crystal of the compound represented by formula (IX)

Chemical formula

[0068] Step 1: Synthesis of Compound 3 Compound 1 (25.8 kg, 200.7 mol), acetonitrile (155 L), pyridine (23.8 kg, 300.9 mol) and Compound 2 (34.5 kg, 220.3 mol) were mixed and stirred at 25 °C for 90 minutes. This reaction solution was added to 25% aqueous ammonia (129.7 kg, 1904.0 mol) and stirred at 25 °C for 270 minutes. The solid was collected by filtration, washed with 50% aqueous acetonitrile, and dried to obtain Compound 3 (30.57 kg, 178.2 mol, yield: 90.2%). HPLC (UV = 254 nm): RT = 7.0 min, HPLC measurement condition 1 1 1H-NMR (DMSO-d6) δ: 6.17 (2H, s), 8.14 - 8.16 (2H, m), 8.38 (1H, d, J = 2.1 Hz), 8.99 (1H, s).

[0069] Step 2: Synthesis of Compound 5 Compound 3 (30.6 kg, 178.3 mol), Compound 4 (40.3 kg, 213.7 mol), N,N-dimethylacetamide (245 L), triethylamine (21.6 kg, 213.5 mol) and ethyl acetate solution of propylphosphonic anhydride (165.6 kg, 267.2 mol) were mixed and stirred at 50 °C for 420 minutes. The reaction solution was cooled to 25 °C, water (92 L) was added, and it was stirred for 150 minutes. The solid was collected by filtration, washed with acetonitrile, and dried to obtain Compound 5 (56.05 kg, 163.8 mol, yield: 92.7%). HPLC (UV = 254 nm): RT = 15.1 min, HPLC measurement condition 1 1 1H-NMR (DMSO-d6) δ: 3.79 (2H, s), 7.31 - 7.35 (1H, m), 7.38 (1H, q, J = 8.7 Hz), 7.56 (1H, dd, J = 7.3, 2.0 Hz), 8.23 (1H, t, J = 2.2 Hz), 8.33 (1H, d, J = 2.1 Hz), 8.64 (1H, d, J = 2.1 Hz), 10.6 (1H, s), 11.1 (1H, s).

[0070] Step 3: Synthesis of Compound 6 Compound 5 (56.1 kg, 164.0 mol), N,N'-carbonyldiimidazole (39.8 kg, 245.5 mol), tetrahydrofuran (561 L), and 1,8-diazabicyclo[5.4.0]-7-undecene (32.4 kg, 212.8 mol) were mixed and stirred at 25 °C for 60 minutes. 7% hydrochloric acid (512.4 kg) was added to this reaction solution, and it was separated into an organic layer and an aqueous layer. Toluene (56 L) and tetrahydrofuran (561 L) were added to the obtained aqueous layer for extraction. The combined organic layers were concentrated under reduced pressure until they reached 56.2 kg, toluene (729 L) and seed crystals of Compound 6 (0.54 kg, 1.3 mol) were added, and the mixture was stirred at 25 °C for 1 day. The slurry was cooled to 5 °C, the solid was collected by filtration, and washed with a mixed solution of cooled tetrahydrofuran (67 L) and toluene (269 L). By drying, Compound 6 (57.22 kg, 138.1 mol, yield: 85.2%) was obtained. HPLC (UV = 254 nm): RT = 9.9 min, HPLC measurement condition 1 1 1H-NMR (DMSO-d6) δ: 2.28 (1.5H, s), 7.12 - 7.18 (1.5H, m), 7.23 - 7.27 (1H, m), 7.31 (1H, br), 7.50 (1H, br), 7.69 (1H, d, J = 7.0 Hz), 8.01 (1H, t, J = 2.1 Hz), 8.50 (1H, br), 8.64 (1H, br), 11.1 (1H, s). As a result of NMR measurement of the obtained solid, 0.5 molecule equivalent of toluene was included, and under reduced pressure drying within the normal operating range, the toluene was not removed. It was confirmed that Compound 6 is a toluene adduct.

[0071] Step 4: Synthesis of Compound 7 Compound 6 (23.6 kg, 57.0 mol), phenyl dichlorophosphate (48.1 kg, 228.2 mol) and sulfolane (104.7 L) were mixed and stirred at 115 °C for 540 minutes. The reaction solution was cooled to 25 °C and added to a solution of potassium acetate (67.0 kg, 684.5 mol), water (203.0 L) and sulfolane (146.7 L) at 50 °C. Sulfolane (20.6 L) and water (105.0 L) were added to the resulting slurry. The slurry was cooled to 25 °C, and the solid was collected by filtration and washed with an aqueous 76% 2-propanol solution. The solid collected by filtration was dissolved in N,N-dimethylacetamide (139.8 L), and 7% hydrochloric acid (21.6 kg) and an aqueous 44% 2-propanol solution (189.0 kg) were added at 50 °C. The slurry was cooled to 25 °C, and the solid was collected by filtration, washed with an aqueous 65% N,N-dimethylacetamide solution and 2-propanol, and then dried to obtain Compound 7 (16.9 kg, 43.6 mol, yield: 76.5%). HPLC (UV = 247 nm): RT = 12.6 min, HPLC measurement condition 2 1 1H-NMR (DMSO-d6) δ: 7.35 - 7.39 (1H, m), 7.48 (1H, m), 7.55 (1H, m), 8.07 (1H, t, J = 4.0 Hz), 8.55 (1H, d, J = 4.0 Hz), 8.71 (1H, d, J = 4.0 Hz), 12.96 (1H, br).

[0072] Step 5: Synthesis of Compound 9 Compound 7 (16.5 kg, 42.7 mol), N,N-dimethylacetamide (115.5 L), N,N-diisopropylethylamine (8.3 kg, 64.0 mol) and Compound 8 (6.1 kg, 51.2 mol) were mixed and stirred at 63 °C for 3 hours. The reaction solution was cooled to 25 °C, acetone (82.5 L) was added, and then 5% hydrochloric acid (55.2 L) was added dropwise over 30 minutes. Water (49.5 L) was added to the resulting slurry, the solid was collected by filtration and washed with 50% aqueous acetone. Ethyl acetate (181.5 L) was added to the obtained solid to dissolve it, and the solution was concentrated to 104.0 kg. Ethyl acetate (82.5 L) was added to the concentrated solution, and the solution was concentrated again to 82.5 kg. The resulting slurry was heated to 55 °C to dissolve it and then cooled to 35 °C. The resulting slurry was stirred for 1 hour, and then heptane (247.5 L) was added. The mixture was cooled to 25 °C, the solid was collected by filtration, washed with a mixed solution of ethyl acetate and heptane, and then dried to obtain Compound 9 (14.98 kg, 35.2 mol, yield: 82.4%). HPLC (UV = 247 nm): RT = 17.2 min, HPLC measurement condition 1 1 1H-NMR (CDCl 3 ) δ: 5.13 (2H, s), 7.23 - 7.24 (2H, m), 7.42 (1H, d, J = 7.3 Hz), 7.67 (1H, t, J = 2.1 Hz), 8.45 (1H, d, J = 2.3 Hz), 8.67 (1H, d, J = 2.3 Hz).

[0073] Step 6: Synthesis of anhydrous crystals of the compound represented by formula (IX) Compound 9 (14.0 kg, 32.9 mol), N,N-dimethylacetamide (77 L), N,N-diisopropylethylamine (10.6 kg, 82.2 mol) and Compound 10 (9.3 kg, 37.8 mol) were mixed and stirred at 50 °C for 2 hours. The reaction solution was cooled to 25 °C, and purified water (28 L) was added. A mixed solution of purified water (42 L), synthetic hydrochloric acid (1.4 kg, 13.2 mol) and 2-propanol (14 L) was added to the resulting slurry, the solid was collected by filtration, washed with 50% aqueous 2-propanol and 2-propanol, and then dried to obtain the undried crystals of the compound represented by formula (IX). Acetone (112 L) was added to the obtained undried crystals and dissolved at 25 °C. The resulting solution was treated with activated carbon, and after the activated carbon was washed with acetone (28 L), purified water (57 L) was added to the treated solution. Purified water (84 L) was added to the resulting slurry, the solid was collected by filtration, washed with 50% 2-propanol water, and dried to obtain anhydrous crystals (15.7 kg, 30.0 mol, yield: 91.3%) of the compound represented by formula (IX). HPLC (UV = 247 nm): RT = 26.8 min, HPLC measurement condition 3 1 H-NMR (CDCl 3 ) δ: 2.75 (4H, t, J = 12.0 Hz), 4.02 (4H, s), 4.74 (2H, s), 7.16 - 7.18 (2H, m), 7.32 - 7.35 (1H, m), 7.65 (1H, t, J = 2.1 Hz), 8.43 (1H, d, J = 2.3 Hz), 8.61 (1H, d, J = 2.3 Hz).

[0074] (Grinding of the anhydrous crystals of the compound represented by formula (IX)) The anhydrous crystals of the compound represented by formula (IX) were sieved through a 1000 μM mesh and then ground under the following conditions. Apparatus: A-O Jet Mill (Seishin Enterprise Co., Ltd.) Feeding method: Feeder Feeding rate: 20 g / hour Grinding pressure: 0.30 MPa Feeding pressure: 0.40 MPa

[0075] (Powder X-ray diffraction experiment of the anhydrous crystals of the compound represented by formula (IX)) The anhydrous crystals of the compound represented by formula (IX) after grinding were subjected to a powder X-ray diffraction experiment under the measurement conditions 2 described above. The powder X-ray diffraction pattern is shown in Figure 3, and the peak list of the powder X-ray diffraction pattern is shown in Figure 4. Hereinafter, in the table of the peak list of the powder X-ray diffraction pattern, Position represents 2θ (°) and Intensity represents the intensity. In the powder X-ray diffraction pattern, peaks were observed at diffraction angles (2θ): 6.5° ± 0.2°, 10.1° ± 0.2°, 13.0° ± 0.2°, 14.1° ± 0.2°, 15.3 ± 0.2°, 15.6° ± 0.2°, 16.2° ± 0.2°, 17.4° ± 0.2°, 18.9° ± 0.2°, 19.9° ± 0.2°, 20.3° ± 0.2°, 21.7° ± 0.2°, 23.0° ± 0.2°, 23.8° ± 0.2°, 25.8° ± 0.2°, 28.8° ± 0.2°, and 30.6° ± 0.2°. The anhydrous crystal of the compound represented by formula (IX) showed characteristic peaks at diffraction angles (2θ): 6.5° ± 0.2°, 15.6° ± 0.2°, 17.4° ± 0.2°, 19.9° ± 0.2°, and 20.3° ± 0.2° in the powder X-ray diffraction pattern.

[0076] (Single crystal structure analysis of the anhydrous crystal of the compound represented by formula (IX)) (Method for preparing single crystal) To 1 mg of the crystal of the compound represented by formula (IX), 400 μL of methanol was added and heated to 50 °C to dissolve it. The solution was dispensed into a 1.5 mL HPLC vial, the lid of the HPLC vial was put on, a syringe needle was pierced into the lid, and it was allowed to stand at room temperature. Single crystals were prepared by the solvent evaporation method. (Single crystal structure analysis) Single crystal diffraction experiments and analyses were carried out by the method described above. Since Cl1 and Cl7C, and H5CA and H6CA are in a disorder relationship, they were analyzed with occupancy ratios of Cl1:Cl7C = 0.75:0.25 and H5CA:H6CA = 0.25:0.75.

[0077] The results of the single crystal structure analysis are shown below. R1 (I > 2.00s(I)) was 0.0555, and it was confirmed that there were no deficiencies or misplacements of electron density from the final difference Fourier.

[0078] The crystallographic data are shown in Table 1.

Table 1

[0079] The fractional atomic coordinates x, y, z (Å×10 4 ) and the equivalent isotropic temperature factor U(eq) (Equivalent Isotropic Displacement Parameters, Å 2 ×10 3 ) of non-hydrogen atoms are shown in Table 2. Here, U(eq) is defined as one-third of the trace of the orthogonalized U ij tensor. Note that the numbers of non-hydrogen atoms in Table 2 correspond to the numbers described in Fig. 5 respectively.

Table 2

[0080] Next, the atomic coordinates x, y, z (Å×10 4 ) and the isotropic temperature factor U(eq) (Isotropic Displacement Parameters, Å 2 ×10 3 ) of hydrogen atoms are shown in Table 3.

Table 3

[0081] The structure in the asymmetric unit of the crystal structure is shown in Fig. 5. Note that the label numbers of non-hydrogen atoms described in Fig. 5 correspond to the numbers of non-hydrogen atoms in Table 2.

[0082] Since only one molecule of the compound represented by formula (IX) was present in the asymmetric unit of the crystal structure, it was identified as an anhydrous crystal of the compound represented by formula (IX).

[0083] The powder X-ray diffraction pattern (λ = 1.5418 Å) calculated from the crystal structure using Mercury (The Cambridge Crystallographic Data Centre, Ver. 4.0.0) was confirmed to be generally consistent with the above powder X-ray diffraction pattern (Figure 3).

[0084] (Differential Scanning Calorimetry of the Anhydrous Crystal of the Compound Represented by Formula (IX)) Approximately 2 mg of the anhydrous crystal of the compound represented by Formula (IX) after pulverization was weighed into an aluminum pan and measured by the method described above. The results are shown in Figure 6. An endothermic peak with an onset temperature of about 261.3 °C was shown.

[0085] (Simultaneous Differential Thermal - Thermogravimetric Measurement of the Anhydrous Crystal of the Compound Represented by Formula (IX)) The anhydrous crystal of the compound represented by Formula (IX) after pulverization was measured by the method described above. The results are shown in Figure 7. An endothermic peak with an onset temperature of about 265.6 °C was shown. Also, no weight loss was confirmed.

[0086] (Raman Spectrum Measurement of the Anhydrous Crystal of the Compound Represented by Formula (IX)) The anhydrous crystal of the compound represented by Formula (IX) after pulverization was subjected to Raman spectrum measurement under the above-described Measurement Condition 1. The results are shown in Figure 8. Also, the main Raman peaks are shown below.

Table 4

[0087] [Example 1A] Synthesis of anhydrous crystals of the compound represented by formula (IX) [Chemical formula] Step 1: Synthesis of Compound 3 Compound 1 (25.00 g, 194.5 mmol), acetonitrile (150.0 mL), pyridine (23.07 g, 291.7 mmol) and Compound 2 (31.97 g, 204.2 mmol) were mixed and stirred at 25 °C for 80 minutes. This reaction solution was added to 28% aqueous ammonia (112.36 g, 1847.4 mmol) and stirred at 25 °C for 5 hours. It was cooled to 5 °C over 20 minutes, the solid was collected by filtration, washed with 50% aqueous acetonitrile, and dried to obtain Compound 3 (32.7 g, 190.6 mmol, yield: 98.0%).

[0088] Step 2: Synthesis of Compound 5 Compound 3 (20.00 g, 116.6 mmol), Compound 4 (24.18 g, 128.2 mmol), N,N-dimethylacetamide (160.0 mL), triethylamine (14.16 g, 139.9 mmol) and 50% ethyl acetate solution of propylphosphonic anhydride (104.59 g, 164.4 mmol) were mixed and stirred at 50 °C for 24 hours. The reaction solution was cooled to 10 °C over 1 hour, water (60.0 mL) was added, and it was stirred for 1 hour. The solid was collected by filtration, washed with acetonitrile, and dried to obtain Compound 5 (36.60 g, 107.0 mmol, yield: 91.8%).

[0089] Step 3: Synthesis of Compound 6 Compound 5 (30.0 g, 87.7 mmol), N,N'-carbodiimidazole (21.3 g, 131.5 mmol), tetrahydrofuran (300.0 mL) and 1,8-diazabicyclo[5,4,0]-7-undecene (17.4 g, 114.0 mmol) were mixed and stirred at 25 °C for 1 hour. To this reaction solution, tetrahydrofuran (255.0 mL), toluene (37.5 mL) and 11% hydrochloric acid (164.8 g, 526.1 mmol) were added for extraction. Water (150.0 mL) was added to the obtained organic layer for further extraction. Sulfolane (90.0 mL) was added to the obtained organic layer and concentrated under reduced pressure until it reached 208.3 g. Toluene (210.0 mL) was added to the concentrated solution and further concentrated under reduced pressure until it reached 206.9 g. Toluene (180 mL) was added to the concentrated solution and further concentrated under reduced pressure until it reached 207.6 g. Toluene (3.0 mL) and seed crystal A of Compound 6 (29.64 mg) were added and stirred at 20 - 30 °C for 19 hours. Toluene (450.0 mL) was added, the slurry was cooled to 5 °C, the crystals were collected by filtration, washed with toluene, and dried to obtain Compound 6 (33.2 g, 80.2 mmol, yield: 91.4%). As a result of NMR measurement of the obtained solid, 0.5 molecule equivalent of toluene was contained and the toluene was not removed under reduced pressure drying within the normal operation range. It was confirmed that Compound 6 is a toluene adduct. 1 1H-NMR (DMSO-d6) δ: 2.30 (1.5H, s), 7.09 - 7.27 (2.5H, m), 7.35 (1H, br), 7.53 (1H, br), 7.72 (1H, d, J = 7.0 Hz), 8.00 (1H, t, J = 2.1 Hz), 8.50 (1H, br), 8.65 (1H, br), 11.1 (1H, s).

[0090] Step 4: Method for synthesizing Compound 7 Compound 6 (55.00 kg, 132.8 mol), phenyl dichlorophosphate (112.0 kg, 530.9 mol) and sulfolane (308.2 kg) were mixed and stirred at 115 °C for 9 hours. The reaction solution was cooled to 50 °C, and after adding sulfolane (123.3 kg), it was added to a solution of potassium acetate (156.4 kg, 1593 mol), water (469 L) and sulfolane (369.8 kg) at 50 °C, and water (244 L) was added to the resulting slurry. The slurry was cooled to 10 °C, the solid was collected by filtration and washed with a 76% aqueous 2-propanol solution. The solid collected by filtration was dissolved in N,N-dimethylacetamide (459.5 kg), and 8% hydrochloric acid (51 kg) and a 44% aqueous 2-propanol solution (436 kg) were added at 50 °C. The resulting slurry was cooled to 5 °C, the solid was collected by filtration, washed with a 76% aqueous 2-propanol solution and 2-propanol, and then dried to obtain Compound 7 (42.65 kg, 110.3 mol, yield: 83.1%).

[0091] Step 5: Method for synthesizing Compound 9 Compound 7 (42.05 kg, 108.8 mol), N,N-dimethylacetamide (276.8 kg), N,N-diisopropylethylamine (21.1 kg, 163 mol) and Compound 8 (15.66 kg, 130.6 mol) were mixed and stirred at 60 °C for 5 hours. The reaction solution was cooled to 25 °C, acetone (165.7 kg) was added, and then 5% hydrochloric acid (143 kg) was added dropwise over about 30 minutes and stirred at 25 °C. Water (126 L) was added to the resulting slurry, the solid was collected by filtration and washed with a 44% acetone-water solution and 2-propanol. Ethyl acetate (151.7 kg) was added to the obtained solid and dissolved at about 50 °C, heptane (57.5 kg) and a seed crystal of Compound 9 (21.0 g, synthesized according to Example 1) were added, and the mixture was stirred at 45 °C. Heptane (402.7 kg) was added and cooled to 0 °C, the solid was collected by filtration, washed with a mixed solution of ethyl acetate and heptane, and then dried to obtain Compound 9 (41.69 kg, 97.95 mol, yield: 90.0%).

[0092] (Alternative method) Synthetic method using cyanomethyl p-toluenesulfonate Compound 7 (4.00 g, 10.35 mmol), N,N-dimethylacetamide (28.0 mL), N,N-diisopropylethylamine (1.68 g, 12.9 mmol) and cyanomethyl p-toluenesulfonate (3.06 g, 14.5 mmol) were mixed and stirred at 60 °C for 6 hours. The reaction solution was cooled to 25 °C, acetone (20.0 mL) was added, and then 2 mol / L hydrochloric acid (9.30 mL, 18.6 mmol) and water (6.0 mL) were added dropwise over about 30 minutes and stirred at 25 °C. Water (6.0 mL) was added to the resulting slurry, the solid was collected by filtration, and washed with 44% aqueous acetone and 2-propanol. Isopropyl acetate (24.0 mL) was added to the obtained solid and dissolved at about 50 °C, heptane (24.0 mL) was added, and stirred at 35 °C. Heptane (56.0 mL) was added, the solid was collected by filtration, washed with a mixed solution of isopropyl acetate and heptane, and dried to obtain Compound 9 (3.32 g, 7.80 mol, yield: 75.4%).

[0093] Step 6: Synthesis of anhydrous crystals of the compound represented by formula (IX) Compound 9 (20.00 g, 47.0 mmol), N,N-dimethylacetamide (100.0 mL), N,N-diisopropylethylamine (15.18 g, 117.5 mmol) and Compound 10 (12.20 g, 49.3 mmol) were mixed and stirred at 60 °C for 4 hours. The reaction solution was cooled to 25 °C over 30 minutes, and purified water (40.0 mL) was added. A mixed solution of purified water (60.0 mL), synthetic hydrochloric acid (1.96 g, 18.8 mmol) and 2-propanol (20.0 mL) was added to the resulting slurry, the solid was collected by filtration, washed with 50% 2-propanol water and 2-propanol, and dried to obtain undried crystals of the compound represented by formula (IX). Acetone (160.0 mL) was added to the obtained undried crystals and dissolved at 25 °C. The resulting solution was treated with activated carbon, the activated carbon was washed with acetone (40.0 mL), purified water (200.0 mL) was added to the treated solution, the solid was collected by filtration, washed with 50% 2-propanol water, and dried to obtain anhydrous crystals of the compound represented by formula (IX) (22.90 g, 43.8 mmol, yield: 93.3%).

Example

[0094] Synthesis of Compound 12

Chem.

[0095] Step 1: Synthesis of Compound 12 Compound 5 (38.6 kg, 112.8 mol), N,N'-carbonyldiimidazole (27.4 kg, 169.0 mol), tetrahydrofuran (375 L) and DBU (22.3 kg, 146.5 mol) were mixed and stirred at 25 °C for 60 minutes. 7% hydrochloric acid (352.5 kg) was added to this reaction solution, and it was separated into an organic layer and an aqueous layer. Toluene (39 L) and tetrahydrofuran (386 L) were added to the obtained aqueous layer for extraction. The combined organic layers were concentrated under reduced pressure until they reached 178.5 kg, toluene (579 L) was added, the slurry was cooled to 5 °C, the solid was collected by filtration, and it was washed with a mixed solution of cooled tetrahydrofuran (46 L) and toluene (186 L). By drying, Compound 12 (37.12 kg, 100.8 mol, yield: 89.4%) was obtained. HPLC (UV = 254 nm): RT = 9.9 min, HPLC measurement condition 1 1 1H-NMR (DMSO-d6) δ: 7.10 (1H, t, J = 9.2 Hz), 7.90 - 7.92 (2H, m), 8.09 (2H, d, J = 7.3 Hz), 8.37 (1H, s), 8.55 (1H, s), 9.90 (1H, s).

Example

[0096] Synthesis of the seed crystal of Compound 6

Chem.

[0097] Step 1: Synthesis of the seed crystal of Compound 6 Compound 12 (1.0 kg, 2.72 mol), toluene (9.0 L) and acetonitrile (1.5 L) were mixed and stirred at 25 °C for 17 hours. The solid was collected by filtration and washed with toluene (5.0 L). By drying, seed crystals of Compound 6 (0.98 kg, 2.37 mol, yield: 87.1%) were obtained. HPLC (UV = 254 nm): RT = 9.9 min, HPLC measurement condition 1 [Example 3A] Synthesis of seed crystal A of Compound 6

Chemical formula

Example

[0098] Synthesis of Compound 7 (alternative method)

Chemical formula

Example

[0099] Synthesis of Compound 5 from Compound 3 using a condensing agent other than T3P (registered trademark)

Chemical formula

Table 5

[0100]

Chemical formula

Table 6

Example

[0101] Synthesis of Compound 12 from Compound 5 Using Bases Other than DBU

Chemical formula

Table 7

Example

[0102] PhPOCl 2 Synthesis of Compound 7 from Compound 6 Using PhPOCl as a Chlorinating Agent

Chemical formula

Table 8

Example

[0103] Synthesis of Compound 7 from Compound 12 using various additives and solvents

Chemical Formula

Table 9

Example

[0104] Synthesis of Compound 7 from Compound 6 using anisole as a solvent in the presence or absence of various additives

Chemical Formula

Table 10

Example

[0105] Regarding Compound 6 obtained by the production method described in Example 1, a powder X-ray diffraction experiment was carried out under the measurement conditions 1 described above. The powder X-ray diffraction pattern is shown in Figure 1, and the peak list of the powder X-ray diffraction pattern is shown in Figure 2. Hereinafter, in the table of the peak list of the powder X-ray diffraction pattern, the height indicates the intensity. In the powder X-ray diffraction pattern, characteristic peaks were observed at diffraction angles (2θ): 9.5 ± 0.2°, 11.8 ± 0.2°, 13.1 ± 0.2°, 14.4 ± 0.2°, 18.4 ± 0.2°, 22.9 ± 0.2°, 23.2 ± 0.2°, 23.7 ± 0.2°, 24.6 ± 0.2°, 26.2 ± 0.2°, and 29.2 ± 0.2°. In the powder X-ray diffraction pattern, characteristic peaks were observed at diffraction angles (2θ): 9.5 ± 0.2°, 11.8 ± 0.2°, 18.4 ± 0.2°, 22.9 ± 0.2°, and 23.2 ± 0.2°.

[0106] Regarding Compound 6 obtained by the production method described in Example 1A, a powder X-ray diffraction experiment was carried out under the measurement conditions 1 described above. The powder X-ray diffraction pattern is shown in Figure 9, and the peak list of the powder X-ray diffraction pattern is shown in Figure 10. Hereinafter, in the table of the peak list of the powder X-ray diffraction pattern, the height indicates the intensity. In the powder X-ray diffraction pattern, characteristic peaks were observed at diffraction angles (2θ): 8.9 ± 0.2°, 18.8 ± 0.2°, 22.3 ± 0.2°, 22.7 ± 0.2°, 23.4 ± 0.2°, 23.9 ± 0.2, 25.1 ± 0.2, 26.1 ± 0.2, 26.4 ± 0.2, 30.3 ± 0.2, 31.3 ± 0.2, and 36.8 ± 0.2°.

[0107] (Reference Example 1) [Chemical formula] Step 1: Synthesis method of Compound 14 Compound 13 (5.50 g, 26.0 mmol), dichloromethane (110.0 mL), diethylamino(difluoro)sulfonium tetrafluoroborate (11.92 g, 52.1 mmol), and triethylamine hydrogen fluoride salt (8.49 mL, 52.1 mmol) were mixed and stirred at 25°C for 21.5 hours. The reaction solution was filtered through Celite (registered trademark), and Celite (registered trademark) was washed with dichloromethane (11.0 mL). The resulting solution was added to a 2 mol / L aqueous sodium hydroxide solution (110.0 mL) at 6°C or lower. The mixture was filtered through Celite (registered trademark), and Celite (registered trademark) was washed with dichloromethane (11.0 mL). The organic layer was separated and concentrated, methanol (16.5 mL) was added to the resulting concentrate, and the mixture was filtered through activated carbon (2.25 g), and the activated carbon was washed with methanol (5.5 mL). Purified water (44.0 mL) was added to the resulting solution, the solid was collected by filtration, washed with 20% methanol-water, and dried to obtain Compound 14 (5.22 g, 22.4 mmol, 86.2%).

[0108] Step 2: Synthesis method of Compound 10 Compound 14 (2.00 g, 8.57 mmol), dichloromethane (10.0 mL) and trifluoroacetic acid (4.89 g, 42.9 mmol) were mixed and stirred at 25 °C for 4 hours. After adding diisopropyl ether (20.0 mL) to the reaction solution, diisopropyl ether (70.0 mL) was added to the resulting slurry and stirred for 2 hours. The solid was collected by filtration, washed with diisopropyl ether, and dried to obtain Compound 10 (1.90 g, 7.69 mmol, 89.7%).

Example

[0109] The following describes a biological test example of the compound represented by formula (IX) produced by the production method according to the present invention. The compound represented by formula (IX) produced by the production method according to the present invention has an inhibitory effect on coronavirus 3CL protease, and any compound that inhibits coronavirus 3CL protease may be used. Specifically, in the evaluation method described below, IC 50 is preferably 50 μM or less, more preferably 1 μM or less, and even more preferably 100 nM or less. EC 50 is preferably 10 μM or less, more preferably 1 μM or less, and even more preferably 100 nM or less.

[0110] Test Example 1: Confirmation test of cytopathic effect (CPE) inhibitory effect using human TMPRSS2 and ACE2-expressing HEK293T cells (HEK293T / ACE2-TMPRSS2 cells) <Operation procedure> · Dilution and dispensing of test samples The test sample was previously diluted to an appropriate concentration with DMSO, a 2- to 5-fold serial dilution series was prepared, and then dispensed into a 384-well plate. · Dilution and dispensing of cells and SARS-CoV-2 HEK293T / ACE2-TMPRSS2 cells (GCP-SL222, 5×10 3 cells / well) and SARS-CoV-2 (200-600 TCID 50 / well) was mixed with the medium (MEM, 2% FBS, penicillin-streptomycin) and dispensed into the wells containing the test samples, and then placed in a CO 2 incubator and cultured for 3 days. · Dispensing of CellTiter-Glo® 2.0 and measurement of luminescence signal After returning the plates cultured for 3 days to room temperature, CellTiter-Glo® 2.0 was dispensed into each well and mixed with a plate mixer. After standing for a certain period of time, the luminescence signal (Lum) was measured with a plate reader. <Calculation of each measurement item value> · 50% SARS-CoV-2 infection cell death inhibition concentration (EC 50 ) calculation When x is the logarithmic value of the compound concentration and y is %Efficacy, the inhibition curve was approximated by the following Logistic regression equation, and the value of x when y = 50 (%) was substituted was calculated as EC 50 as. y = min + (max - min) / {1 + (X50 / x) ^Hill} %Efficacy = {(Sample - virus control) / (cell control - virus control)} * 100% cell control: the average of Lum of cell control wells virus control: the average of Lum of virus control wells min: lower limit value of the y-axis, max: upper limit value of the y-axis, X50: x-coordinate of the inflection point, Hill: slope of the curve at the midpoint between min and max

[0111] The compound according to the present invention was tested essentially as described above. EC 50 values are shown below. (Results) Compound represented by formula (IX): 1.66 nM

[0112] Test Example 2: Inhibitory Activity Test against SARS-CoV-2 3CL Protease <Materials> · Commercially available Recombinant SARS-CoV-2 3CL Protease · Commercially available substrate peptide Dabcyl-Lys-Thr-Ser-Ala-Val-Leu-Gln-Ser-Gly-Phe-Arg-Lys-Met-Glu(Edans)-NH 2 (SEQ ID NO: 1) · Internal Standard peptide Dabcyl-Lys-Thr-Ser-Ala-Val-Leu( 13 C 6 , 15 N)-Gln (SEQ ID NO: 2) Dabcyl-Lys-Thr-Ser-Ala-Val-Leu( 13 C 6 , 15 N)-Gln can be synthesized with reference to the literature (Atherton, E.; Sheppard, R. C., “In Solid Phase Peptide Synthesis, A Practical Approach”, IRL Press at Oxford University Press, 1989 and Bioorg. Med. Chem., Vol. 5, No. 9, 1997, pp. 1883-1891, etc.). An example is shown below. Using Rink amide resin, by Fmoc solid-phase synthesis, H-Lys-Thr-Ser-Ala-Val-Leu( 13 C 6 , 15Synthesize N)-Glu(resin)-OαOtBu (the Lys side chain is protected by Boc, the Thr side chain is protected by a tert-butyl group, the Ser side chain is protected by a tert-butyl group, the C-terminal OH of Glu is protected by a tert-butyl group, and the carboxylic acid of the Glu side chain is condensed with the resin). The modification of the N-terminal Dabcyl group is carried out by condensing 4-dimethylaminoazobenzene-4'-carboxylic acid (Dabcyl-OH) on the resin using EDC / HOBT. Final deprotection and cleavage from the resin are performed by treatment with TFA / EDT = 95:5. Then, it is purified by reverse-phase HPLC. ·RapidFire Cartridge C4 typeA <Operation procedure> ·Preparation of assay buffer In this test, an assay buffer consisting of 20 mM Tris-HCl, 1 mM EDTA, 10 mM DTT, and 0.01% BSA is used. ·Dilution and dispensing of test samples Dilute the test sample appropriately with DMSO in advance, prepare a 2 - 5-fold serial dilution series, and then dispense it into a 384-well plate. ·Addition of enzyme and substrate, enzyme reaction Add 8 μM of the substrate and 6 nM or 0.6 nM of the enzyme solution to the prepared compound plate, and incubate at room temperature for 3 - 5 hours. Then, add the reaction stop solution (0.067 μM Internal Standard, 0.1% formic acid, 10% or 25% acetonitrile) to stop the enzyme reaction. ·Measurement of reaction products The completed reaction plate is measured using a RapidFire System 360 and a mass spectrometer (Agilent, 6550 iFunnel Q-TOF), or a Rapid Fire System 365 and a mass spectrometer (Agilent, 6495C Triple Quadrupole). As the mobile phase during measurement, use solution A (75% isopropanol, 15% acetonitrile, 5 mM ammonium formate) and solution B (0.01% trifluoroacetic acid, 0.09% formic acid). The reaction products detected by the mass analyzer are calculated using RapidFire Integrator or a program capable of equivalent analysis to obtain the Product area value. At the same time, the Internal Standard detected simultaneously is also calculated to obtain the Internal Standard area value. <Calculation of each measurement item value> ·Calculation of P / IS Calculate P / IS by using the area value obtained in the previous item according to the following formula. P / IS = Product area value / Internal Standard area value ·50% SARS-CoV-2 3CL protease inhibitory concentration (IC 50 ) calculation When x is the logarithmic value of the compound concentration and y is %Inhibition, approximate the inhibition curve with the following Logistic regression equation, and calculate the value of x when y = 50 (%) as IC 50 and calculate it as. y = min + (max - min) / {1 + (X50 / x) ^Hill} %Inhibition = {1-(Sample - Control(-)) / Control(+)-Control(-))} * 100 Control(-): the average of P / IS of enzyme inhibited condition wells Control(+): the average of P / IS of DMSO control wells min: lower limit value of the y-axis, max: upper limit value of the y-axis, X50: x-coordinate of the inflection point, Hill: slope of the curve at the midpoint between min and max

[0113] The compound represented by formula (IX) produced by the production method according to the present invention was tested essentially as described above. IC 50 values are shown below. (Result) The compound represented by formula (IX): 0.00036 μM

[0114] The following formulation examples are merely illustrative and are not intended to limit the scope of the invention in any way. The compound represented by formula (IX) produced by the production method according to the present invention can be administered as a pharmaceutical composition by any conventional route, particularly enterally, for example, orally, in the form of tablets or capsules, or parenterally, for example, in the form of injection solutions or suspensions, topically, for example, in the form of lotions, gels, ointments or creams, or nasally or in suppository form. A pharmaceutical composition containing the compound represented by formula (IX) produced by the production method according to the present invention in free form or in the form of a pharmaceutically acceptable salt, together with at least one pharmaceutically acceptable carrier or diluent, can be produced by conventional methods, by mixing, granulating or coating methods. For example, oral compositions can be made into tablets, granules, capsules containing excipients, disintegrants, binders, lubricants, etc. and the active ingredient, etc. Also, injection compositions can be made into solutions or suspensions, may be sterilized, and may contain preservatives, stabilizers, buffering agents, etc.

Industrial Applicability

[0115] The compound represented by formula (IX) produced by the production method according to the present invention has an inhibitory effect on coronavirus 3CL protease and is considered useful as a therapeutic agent and / or prophylactic agent for diseases or conditions involving coronavirus 3CL protease. The novel synthetic intermediates according to the present invention or their salts, and the production method according to the present invention are useful in the manufacture of pharmaceuticals.

Claims

1. Formula (I): 【Chemistry 1】 (In the formula, R １ is halogen, C1-C4 alkyl, haloC1-C4 alkyl or cyano, and Z is CH or N.) or a salt thereof with a compound represented by formula (II): 【Chemistry 2】 (In the formula, R ２ are each independently halogen, haloC1-C4 alkyl or cyano, and n is an integer of 1 to 5.) or a salt thereof is reacted with T3P (registered trademark), ethylphosphonic anhydride, n-butylphosphonic anhydride, PyBOP (registered trademark), HATU, WSCD, WSCD.HCl, MsCl and ClP(O)(OPh) in the presence or absence of a base. ２ The compound of formula (III): 【Chemistry 3】 (wherein the symbols are as defined above) or a salt thereof.

2. The process according to claim 1, wherein the condensing agent is T3P (registered trademark).

3. The process according to claim 1 or 2, which is carried out in the presence of triethylamine.

4. The compound represented by formula (III) may be a compound represented by formula (IV): 【Chemistry 4】 The method according to claim 3, wherein the compound is represented by the formula:

5. According to the method of claim 4, a compound represented by the formula (IV): 【Chemistry 5】 or a salt thereof, comprising the step of obtaining a compound represented by formula (IX): 【Chemistry 6】 A method for producing a compound represented by the formula:

6. According to the method of claim 4, a compound represented by formula (IV): 【Chemistry 7】 or a salt thereof; Formula (IV): 【Chemistry 8】 or a salt thereof with N,N'-carbonyldiimidazole in the presence of a base to obtain a compound represented by the formula (VI): 【Chemistry 9】 or a salt thereof, or a solvate thereof; and formula (VI): 【Chemistry 10】 or a salt thereof, or a solvate thereof, is reacted with a chlorinating agent selected from the group consisting of phosphorus oxychloride, phenyl dichlorophosphate, and phenylphosphonic acid dichloride, in the presence of water and sulfolane, and in the presence or absence of a solvent, to obtain a compound represented by the formula (VIII): 【Chemistry 11】 or a salt thereof, comprising the steps of: 【Chemistry 12】 A method for producing a compound represented by the formula:

7. The method according to claim 6, wherein the base used in the step of producing compound (VI) is DBU.

8. The method according to claim 6, wherein the chlorinating agent used in the process for producing compound (VIII) is phosphorus oxychloride or phenyl dichlorophosphate.

9. Formula (IV): 【Chemistry 13】 or a salt thereof.

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