WO2023136277A1 - Compound and use of same - Google Patents

Abstract

According to the present invention, a protease inhibitor contains, as an active ingredient, a compound represented by general formula (I). In general formula (I), R11 and R12 each independently represent a hydrogen atom or a group represented by general formula (II), and when either one of R11 and R12 is a hydrogen atom, the other is a group represented by general formula (II). R13 represents a 1H-imidazol-4-yl group or a pyrrolidin-2-on-3-yl group, R14 represents a formyl group, -CH=CHCO2R', -C(=O)-R", or a cyano group, R' represents a substituted or unsubstituted alkyl group having 1-4 carbon atoms, or a benzyl group, and R" represents a substituted or unsubstituted alkyl group having 1-4 carbon atoms.

Description

Compounds and uses thereof

This disclosure relates to compounds and uses thereof. Specifically, the present disclosure relates to novel compounds and protease inhibitors and antiviral pharmaceutical compositions comprising said novel compounds. The present disclosure claims priority based on Japanese Patent Application No. 2022-002869 filed in Japan on January 12, 2022, the content of which is incorporated herein.

Novel coronavirus infection (COVID-19; Coronavirus disease 2019) is caused by severe acute respiratory syndrome (Severe acute respiratory syndrome) coronavirus 2 (SARS-CoV-2) (hereinafter referred to as "SARS coronavirus-2") It is an infection caused by Drugs that block the replication of SARS-coronavirus-2 and halt the progression of viral infection in patients are being sought as therapeutics for COVID-19. So far, the main protease (M ^pro ) of SARS-coronavirus-2 has been identified as a potential target for therapeutic agents. M ^pro is a cysteine protease and an enzyme involved in viral replication. 3C-like protease, also called C30 endopeptidase.

Rupintrivir and the like are known as inhibitory compounds against M ^pro , and their effectiveness against SARS-coronavirus-2 is being investigated. However, most inhibitory compounds against M ^pro are peptide-like compounds in which a structure in which α-amino acids are polymerized is bound to the reactive site (warhead) (see, for example, Non-Patent Document 1, etc.).

As described above, the known inhibitory compounds against M ^pro are peptide-like compounds, so from the viewpoint of pharmacokinetics and future resistance mutant strains, compounds that exhibit inhibitory activity that is necessarily satisfactory in vivo. I can't say. Thus, there is a need for non-peptidomimetic compounds with inhibitory activity against M ^pro .

Pillaiyar T et al., “An Overview of Severe Acute Respiratory Syndrome-Coronavirus (SARS-CoV) 3CL Protease Inhibitors: Peptidomimetics and Small Molecule Chemotherapy.”, Journal of Medicinal Chemistry, Vol. 59, Issue 14, pp. 659 5-6628 , 2016.

The present disclosure has been made in view of the above circumstances, a novel compound having inhibitory activity against protease and having a non-peptide-like structure, and a protease inhibitor and an antiviral drug containing the novel compound A composition is provided.

As a result of extensive studies to achieve the above object, the inventors have found that a benzoic acid amide compound having a warhead portion with a specific structure exhibits excellent inhibitory activity against proteases, and have completed the present disclosure. reached.

That is, the present disclosure includes the following aspects.
(1) A protease inhibitor containing a compound represented by the following general formula (I) as an active ingredient.

In general formula (I), R ¹¹ and R ¹² are each independently a hydrogen atom or a group represented by general formula (II) below, and when either one of R ¹¹ and R ¹² is a hydrogen atom, The other is a group represented by the following general formula (II). R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group, R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, —C(═O)—R″, or a cyano group, R′ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms or a benzyl group, and R″ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms. .

In general formula (II), an asterisk represents a binding site to a benzene ring, R ²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted or a group represented by the following general formula (III).

In general formula (III), an asterisk represents a binding site with an amide bond, Y ³¹ is a methylene group or a methylmethylene group, Q ³¹ is -NH- or -NH-C(=O)- and R ³¹ is a substituted or unsubstituted cyclohexyl group or a substituted or unsubstituted phenyl group.

(2) The protease inhibitor according to (1), which contains a compound represented by the following general formula (I-1) or a compound represented by the following general formula (I-2) as an active ingredient.

In general formula (I-1), R ¹¹¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyrrolidinyl group, or the general formula (III) is a group represented by

In general formula (I-2), R ¹²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyrrolidinyl group, or the general formula (III) is a group represented by

(3) The protease inhibitor according to claim 1 or 2, which contains a compound represented by the following formula (I-1-1) as an active ingredient.

(4) The protease inhibitor according to any one of (1) to (3), wherein the protease is a cysteine protease.
(5) The protease inhibitor according to any one of (1) to (4), wherein the protease is 3C protease or 3C-like protease.
(6) The protease inhibitor according to any one of (1) to (5), wherein the protease is a 3C-like protease of SARS-coronavirus-2.
(7) An antiviral pharmaceutical composition comprising the protease inhibitor according to any one of (1) to (6) and a pharmaceutically acceptable carrier.
(8) A compound represented by the following general formula (I).

In general formula (I), R ¹¹ and R ¹² are each independently a hydrogen atom or a group represented by general formula (II) below, and when either one of R ¹¹ and R ¹² is a hydrogen atom, The other is a group represented by the following general formula (II). R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group, R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, —C(═O)—R″, or a cyano group, R′ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms or a benzyl group, and R″ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms. .

In general formula (II), an asterisk represents a binding site with an amide bond, R ²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted or a group represented by the following general formula (III).

In general formula (III), an asterisk represents a binding site with an amide bond, Y ³¹ is a methylene group or a methylmethylene group, Q ³¹ is -NH- or -NH-C(=O)- and R ³¹ is a substituted or unsubstituted cyclohexyl group or a substituted or unsubstituted phenyl group.

≪Protease inhibitor≫
The protease inhibitor of this embodiment contains a compound represented by the following general formula (I) (hereinafter sometimes referred to as "compound (I)") as an active ingredient.

In general formula (I), R ¹¹ and R ¹² are each independently a hydrogen atom or a group represented by general formula (II) below, and when either one of R ¹¹ and R ¹² is a hydrogen atom, The other is a group represented by the following general formula (II). R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group, R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, —C(═O)—R″, or a cyano group, R′ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms or a benzyl group, and R″ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms. .

In general formula (II), an asterisk represents a binding site to a benzene ring, R ²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted or a group represented by the following general formula (III).

In general formula (III), an asterisk represents a bonding site with a benzene ring, Y ³¹ is a methylene group or a methylmethylene group, Q ³¹ is -NH- or -NH-C(=O)- and R ³¹ is a substituted or unsubstituted cyclohexyl group or a substituted or unsubstituted phenyl group.

The protease inhibitor of the present embodiment can effectively inhibit proteases by containing the compound (I).
In addition, "containing as an active ingredient" means containing an effective amount of compound (I) for inhibition of protease.

The protease to be inhibited by the protease inhibitor of the present embodiment is not particularly limited, but examples include serine protease, cysteine protease, threonine protease, aspartic protease, glutamic protease, metalloprotease, asparagine protease, and the like. Among them, cysteine protease is preferred.

Cysteine proteases, also called thiol proteases and SH proteases, are proteases that use the thiol group of cysteine as a nucleophilic group at the catalytic site. Specific examples of cysteine proteases include papain, ficin, bromelain, etc. for plants, cathepsin B, cathepsin H, cathepsin L, calcium-dependent protease, etc. for animals, and 3C protease for viruses. , 3C-like (3CL) proteases, and the like.

Among them, the cysteine protease is preferably a 3C protease or a 3C-like (3CL) protease, more preferably a 3C-like (3CL) protease. 3C protease and 3CL protease are cysteine proteases possessed by viruses that cause various infectious diseases, and are known as proteases involved in virus replication. Therefore, inhibitors of 3C protease or 3CL protease can suppress viral replication and proliferation, and are useful as therapeutic agents for infectious diseases caused by the virus. As shown in the examples below, compound (I) has a particularly excellent inhibitory activity against the 3CL protease (M ^pro ) of SARS-coronavirus-2. As a protease of interest, the 3CL protease of SARS-coronavirus-2 (M ^pro ) is particularly preferred.

Next, the structure of compound (I), which is an active ingredient, and its production method will be explained in detail.

<Compound (I)>
Compound (I) is a compound represented by the following general formula (I). In compound (I), R ¹³ and R ¹⁴ correspond to the warhead portion, in particular R ¹³ interacts with histidine residue at position 163 in 3CL protease of SARS-coronavirus-2, and R ¹⁴ is SARS -It can specifically bind to 3CL protease of coronavirus-2 by interacting with the 145th cysteine residue in 3CL protease of coronavirus-2. In addition, since the ortho- or meta-substituted aminobenzoic acid moiety in compound (I) is not an amino acid structure constituting a protein, it is highly resistant to degradation in vivo and can exhibit stable pharmacokinetics.

The relative arrangement of the histidine and cysteine residues, which are essential for activity expression in SARS-coronavirus-2 3CL protease, is also preserved in cysteine proteases other than SARS-coronavirus-2 3CL protease. ing. Therefore, compound (I) can also specifically bind to cysteine proteases other than the 3CL protease of SARS-coronavirus-2 exemplified above.

In general formula (I), R ¹¹ and R ¹² are each independently a hydrogen atom or a group represented by general formula (II) below, and when either one of R ¹¹ and R ¹² is a hydrogen atom, The other is a group represented by the following general formula (II). R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group, R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, —C(═O)—R″, or a cyano group, R′ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms or a benzyl group, and R″ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms. .

In general formula (II), an asterisk represents a binding site to a benzene ring, R ²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted or a group represented by the following general formula (III).

In general formula (III), an asterisk represents a bonding site with a benzene ring, Y ³¹ is a methylene group or a methylmethylene group, Q ³¹ is -NH- or -NH-C(=O)- and R ³¹ is a substituted or unsubstituted cyclohexyl group or a substituted or unsubstituted phenyl group.

[R ¹¹ and R ¹² ]
In general formula (I), R ¹¹ and R ¹² are each independently a hydrogen atom or a group represented by general formula (II) below (hereinafter sometimes referred to as “group (II)”). When either one of R ¹¹ and R ¹² is a hydrogen atom, the other is group (II).

(Group (II))
Group (II) is represented by the following general formula (II). In general formula (II), an asterisk represents a binding site with a benzene ring.

(1) ^R21
R ²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyrrolidinyl group, or a group represented by the following general formula (III) (hereinafter, " (sometimes referred to as "group (III)").

In general formula (III), an asterisk represents a bonding site with a benzene ring, Y ³¹ is a methylene group or a methylmethylene group, Q ³¹ is -NH- or -NH-C(=O)- and R ³¹ is a substituted or unsubstituted cyclohexyl group or a substituted or unsubstituted phenyl group.

Examples of the alkyl group having 1 to 3 carbon atoms for R ²¹ include an ethyl group, a methyl group, a propyl group and an isopropyl group.

Examples of substituents of the alkyl group for R ²¹ include a halogen atom, an aryl group having 6 to 12 carbon atoms, a halogenated aryl group having 6 to 12 carbon atoms, a heteroaryl group having 2 to 12 carbon atoms, a carbon A halogenated heteroaryl group having a number of 2 or more and 12 or less and the like can be mentioned.

The halogen atom includes, for example, fluorine atom, chlorine atom, bromine atom, iodine atom and the like.

Examples of the aryl group having 6 to 12 carbon atoms include a phenyl group, 2-methylphenyl group, 3-methylphenyl group, 4-methylphenyl group, 2-naphthyl group and biphenyl group.

Examples of the halogenated aryl group having 6 or more and 12 or less carbon atoms include a 4-chlorophenyl group and the like.

Examples of the heteroaryl group having 2 or more and 12 or less carbon atoms include a 5-indolyl group and a 2-pyridyl group.

Examples of the halogenated heteroaryl group having 2 to 12 carbon atoms include 2-chlorothiazol-4-yl group and the like.

The alkyl group for R ²¹ may be substituted with one substituent, or may be substituted with a combination of two or more substituents.

Examples of substituents of the phenyl group and pyrrolidinyl group in R ²¹ include a halogen atom, an alkyl group having 1 to 3 carbon atoms, an aryl group having 6 to 12 carbon atoms, a heteroaryl group having 2 to 12 carbon atoms, Aralkyl groups having 7 to 12 carbon atoms, heteroaralkyl groups having 4 to 12 carbon atoms, piperazin-1-ylsulfonyl groups, alkylpiperazin-1-ylsulfonyl groups and the like can be mentioned.

Examples of the halogen atom, the aryl group having 6 to 12 carbon atoms, and the heteroaryl group having 2 to 12 carbon atoms are the same as those exemplified for the alkyl group substituent for R ²¹ above.

Examples of the alkyl group having 1 to 3 carbon atoms are the same as those exemplified as the alkyl group for R ²¹ above.

The aralkyl group having 7 or more and 12 or less carbon atoms includes, for example, a benzyl group.

Examples of the heteroaralkyl group having 4 to 12 carbon atoms include 5-indolylmethyl group and the like.

Examples of alkylpiperazin-1-ylsulfonyl groups include 4-methylpiperazin-1-ylsulfonyl groups.

The phenyl group and pyrrolidinyl group in R ²¹ may each be substituted with one type of substituent, or may be substituted with a combination of two or more types of substituents.

Among them, R ²¹ is an alkyl group having 1 to 3 carbon atoms substituted with an aryl group having 6 to 12 carbon atoms, a halogen atom, a piperazin-1-ylsulfonyl group or an alkylpiperazin-1-ylsulfonyl group. A substituted phenyl group, a pyrrolidinyl group substituted with an aralkyl group having 7 to 12 carbon atoms or a heteroaralkyl group having 4 to 12 carbon atoms, or group (III) is preferred, and a methyl group substituted with a biphenyl group ( that is, a biphenylmethyl group), a phenyl group substituted with a fluorine atom or a 4-methylpiperazin-1-ylsulfonyl group, a pyrrolidinyl group substituted with a benzyl group or a 5-indolylmethyl group, or the group (III) is more preferable.

(1-1) group (III)
In general formula (III), Y ³¹ is a methylene group or a methylmethylene group.

In general formula (III), Q ³¹ is -NH- or -NH-C(=O)-.

In general formula (III), R ³¹ is a substituted or unsubstituted cyclohexyl group or a substituted or unsubstituted phenyl group.

Substituents of the cyclohexyl group in R ³¹ include a halogen atom, an alkyl group having 1 to 3 carbon atoms, an aryl group having 6 to 12 carbon atoms, a piperazin-1-ylsulfonyl group, and an alkylpiperazin-1-ylsulfonyl group. etc. Examples of the halogen atom and the aryl group having 6 to 12 carbon atoms are the same as those exemplified for the substituent of the alkyl group for R ²¹ above. Examples of the alkyl group having 1 to 3 carbon atoms are the same as those exemplified as the alkyl group for R ²¹ above.

Examples of the substituent of the phenyl group for R ³¹ include the same as those exemplified as the substituent of the phenyl group for R ²¹ above.

The cyclohexyl group and phenyl group for R ³¹ may each be substituted with one type of substituent, or may be substituted by a combination of two or more types of substituents.

Among them, R ³¹ is preferably an unsubstituted cyclohexyl group or a substituted phenyl group, more preferably an unsubstituted cyclohexyl group or a phenyl group substituted with a halogen atom, and an unsubstituted cyclohexyl group or a fluorine atom-substituted Further preferred are phenyl groups.

A preferable combination of Y ³¹ , Q ³¹ and R ³¹ in group (III) is that Y ³¹ is a methylene group, Q ³¹ is -NH- or -NH-C(=O)-, and a combination in which R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, Q ³¹ is -NH-C(=O)-, and R ³¹ is a phenyl group substituted with a halogen atom; A certain combination etc. are mentioned. These combinations are merely examples of preferred combinations of Y ³¹ , Q ³¹ and R ³¹ , and preferred combinations of Y ³¹ , Q ³¹ and R ³¹ are not limited to these.

[R ¹³ ]
In general formula (I), R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group.

[ ^R14 ]
In general formula (I), R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, —C(═O)—R″, or a cyano group, and R′ is a substituted or unsubstituted carbon number of 1 or more. 4 or less alkyl group or benzyl group, and R″ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms.

(R' and R'')
Examples of the alkyl group having 1 to 4 carbon atoms in R′ and R″ include methyl group, ethyl group, propyl group, isopropyl group, butyl group, sec-butyl group, isobutyl group and tert-butyl group. be done.

Examples of substituents of the alkyl groups in R′ and R″ include halogen atoms, aryl groups having 6 to 12 carbon atoms, and the like. Halogen atoms and aryl groups having 6 to 12 carbon atoms include the above Examples thereof are the same as those exemplified for the substituent of the alkyl group for R ²¹ .

The alkyl groups in R' and R'' may be substituted with one type of substituent, or may be substituted with a combination of two or more types of substituents.

Among them, R' is preferably a methyl group, an ethyl group, or a benzyl group, and more preferably an ethyl group or a benzyl group.

In addition, R" is preferably a methyl group or an ethyl group, more preferably an ethyl group.

Preferred compounds (I) include, for example,
R ¹¹ is group (II), and R ²¹ in group (II) is an alkyl group having 1 to 3 carbon atoms substituted with an aryl group having 6 to 12 carbon atoms, a halogen atom, piperazine-1 - A phenyl group substituted with an ylsulfonyl group or an alkylpiperazin-1-ylsulfonyl group, a pyrrolidinyl group substituted with an aralkyl group having 7 to 12 carbon atoms or a heteroaralkyl group having 4 to 12 carbon atoms, or a group ( III) is;
R ¹² is a hydrogen atom;
R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group; and
R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, or a cyano group;
etc.

Alternatively, preferred compounds (I) include, for example,
R ¹¹ is a hydrogen atom;
R ¹² is group (II), and R ²¹ in group (II) is an alkyl group having 1 to 3 carbon atoms substituted with an aryl group having 6 to 12 carbon atoms, a halogen atom, piperazine-1 - A phenyl group substituted with an ylsulfonyl group or an alkylpiperazin-1-ylsulfonyl group, a pyrrolidinyl group substituted with an aralkyl group having 7 to 12 carbon atoms or a heteroaralkyl group having 4 to 12 carbon atoms, or a group ( III) is;
R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group; and
R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, or a cyano group;
etc.

More preferred compounds (I) include, for example,
R ¹¹ is a group (II), and R ²¹ in the group (II) is a methyl group substituted with a biphenyl group (that is, a biphenylmethyl group), a fluorine atom, or a 4-methylpiperazin-1-ylsulfonyl group a phenyl group substituted with, a benzyl group or a pyrrolidinyl group substituted with a 5-indolylmethyl group, or a group (III) (in group (III), Y ³¹ is a methylene group, Q ³¹ is —NH—, or a combination of -NH-C(=O)- and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group and Q ³¹ is -NH-C(=O)- and R ³¹ is a phenyl group substituted with a halogen atom);
R ¹² is a hydrogen atom;
R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group; and
R ¹⁴ is a formyl group, -CH=CHCO ₂ R'(R' is an ethyl group or a benzyl group), or a cyano group;
etc.

Alternatively, more preferred compounds (I) include, for example,
R ¹¹ is a hydrogen atom;
R ¹² is a group (II), and R ²¹ in the group (II) is a methyl group substituted with a biphenyl group (that is, a biphenylmethyl group), a fluorine atom, or a 4-methylpiperazin-1-ylsulfonyl group a phenyl group substituted with, a benzyl group or a pyrrolidinyl group substituted with a 5-indolylmethyl group, or a group (III) (in group (III), Y ³¹ is a methylene group, Q ³¹ is —NH—, or a combination of -NH-C(=O)- and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group and Q ³¹ is -NH-C(=O)- and R ³¹ is a phenyl group substituted with a halogen atom);
R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group; and
R ¹⁴ is a formyl group, -CH=CHCO ₂ R'(R' is an ethyl group or a benzyl group), or a cyano group;
etc.

More specifically, preferred compounds (I) include, for example, compounds represented by the following general formula (I-1) (hereinafter sometimes referred to as "compound (I-1)"), the following general formula (I -2) (hereinafter sometimes referred to as "compound (I-2)"), a compound represented by the following general formula (1-3a) (hereinafter, "compound (I-3a)" may be referred to as), a compound represented by the following general formula (1-3b) (hereinafter sometimes referred to as "compound (I-3b)"), represented by the following general formula (1-4a) a compound (hereinafter sometimes referred to as "compound (I-4a)"), a compound represented by the following general formula (1-4b) (hereinafter sometimes referred to as "compound (I-4b)"), A compound represented by the following general formula (1-5a) (hereinafter sometimes referred to as "compound (I-5a)"), a compound represented by the following general formula (1-5b) (hereinafter referred to as "compound ( I-5b)”), the compound represented by the following general formula (1-6a) (hereinafter sometimes referred to as “compound (I-6a)”), the following general formula (1-6b ) (hereinafter sometimes referred to as “compound (I-6b)”) and the like. These compounds are merely examples of preferable compound (I), and preferable compound (I) is not limited to these.

In general formula (I-1), R ¹¹¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyrrolidinyl group, or the general formula (III) is a group represented by

In general formula (I-2), R ¹²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyrrolidinyl group, or the general formula (III) is a group represented by

In general formulas (I-3a) to (I-3b), R ¹³¹ and R ¹³² are each independently a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or It is an unsubstituted pyrrolidinyl group or a group represented by the general formula (III).

In general formulas (I-4a) to (I-4b), R ¹⁴¹ and R ¹⁴² are each independently a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or It is an unsubstituted pyrrolidinyl group or a group represented by the general formula (III).

In general formulas (I-5a) to (I-5b), R ¹⁵¹ and R ¹⁵² are each independently a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or It is an unsubstituted pyrrolidinyl group or a group represented by the general formula (III).

In general formulas (I-6a) to (I-6b), R ¹⁶¹ and R ¹⁶² are each independently a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or It is an unsubstituted pyrrolidinyl group or a group represented by the general formula (III).

[R ¹¹¹ , R ¹²¹ , R ¹³¹ , R ¹³² , R ¹⁴¹ , R ¹⁴² , R ¹⁵¹ , R ¹⁵² , R ¹⁶¹ and R ¹⁶² ]
R ¹¹¹ , R ¹²¹ , R ¹³¹ , R ¹³² , R ¹⁴¹ , R ¹⁴² , R ¹⁵¹ , R ¹⁵² , R ¹⁶¹ and R ¹⁶² are each independently a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, substituted Alternatively, it is an unsubstituted phenyl group, a substituted or unsubstituted pyrrolidinyl group, or group (III). The group (III) here includes the same groups as those exemplified for the above "R ²¹ ".

Among them, R ¹¹¹ , R ¹²¹ , R ¹³¹ , R ¹³² , R ¹⁴¹ , R ¹⁴² , R ¹⁵¹ , R ¹⁵² , R ¹⁶¹ and R ¹⁶² are each independently substituted with an aryl group having 6 to 12 carbon atoms. an alkyl group having 1 to 3 carbon atoms, a halogen atom, a phenyl group substituted with a piperazin-1-ylsulfonyl group or an alkylpiperazin-1-ylsulfonyl group, an aralkyl group having 7 to 12 carbon atoms, or 4 carbon atoms A pyrrolidinyl group substituted with a heteroaralkyl group of 12 or less, or group (III) is preferable, and a methyl group substituted with a biphenyl group (that is, a biphenylmethyl group), a fluorine atom or 4-methylpiperazin-1-ylsulfonyl A phenyl group substituted with a group, a pyrrolidinyl group substituted with a benzyl group or a 5-indolylmethyl group, or group (III) is more preferred.

Preferred compounds (I-1) include, for example, R ¹¹¹ is a methyl group substituted with a biphenyl group (that is, a biphenylmethyl group), a fluorine atom or a phenyl substituted with a 4-methylpiperazin-1-ylsulfonyl group a pyrrolidinyl group substituted with a benzyl group or a 5-indolylmethyl group, or the group (III) (wherein Y ³¹ is a methylene group and Q ³¹ is —NH— or —NH—C (=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, Q ³¹ is -NH-C(=O)-, and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-1) include, for example, compounds represented by the following formulas (I-1-1) to (I-1-6) (hereinafter, “compound (I-1-1)” etc.) and the like. These compounds are merely examples of preferable compound (I-1), and preferable compound (I-1) is not limited to these.

Preferred compounds (I-2) include, for example, a phenyl group in which R ¹²¹ is substituted with a fluorine atom or a 4-methylpiperazin-1-ylsulfonyl group, or group (III) (in group (III), Y ³¹ is a methylene group, Q ³¹ is -NH- or -NH-C(=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, and Q ³¹ is -NH-C(=O)- and R ³¹ is a phenyl group substituted with a halogen atom).

Specific examples of preferred compounds (I-2) include compounds represented by the following formulas (I-2-1) to (I-2-3) (hereinafter, “compound (I-2-1)” etc.) and the like. These compounds are merely examples of preferable compound (I-2), and preferable compound (I-2) is not limited to these.

Preferred compounds (I-3a) include, for example, a methyl group substituted with a ^biphenyl group (that is, a biphenylmethyl group), a fluorine atom or a phenyl group substituted with a 4-methylpiperazin-1-ylsulfonyl group. a pyrrolidinyl group substituted with a benzyl group or a 5-indolylmethyl group, or the group (III) (wherein Y ³¹ is a methylene group and Q ³¹ is —NH— or —NH—C (=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, Q ³¹ is -NH-C(=O)-, and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-3a) include, for example, compounds represented by the following formulas (I-3a-1) to (I-3a-5) (hereinafter, “compound (I-3a-1)” etc.) and the like. These compounds are merely examples of preferred compounds (I-3a), and preferred compounds (I-3a) are not limited to these.

Preferred compounds (I-3b) include, for example, a phenyl group in which R ¹³² is substituted with a fluorine atom or a 4-methylpiperazin-1-ylsulfonyl group, or group (III) (in group (III), Y ³¹ is a methylene group, Q ³¹ is -NH- or -NH-C(=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, and Q ³¹ is -NH-C(=O)- and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-3b) include, for example, compounds represented by the following formulas (I-3b-1) to (I-3b-3) (hereinafter, “compound (I-3b-1)” etc.) and the like. These compounds are merely examples of preferred compounds (I-3b), and preferred compounds (I-3b) are not limited to these.

Preferred compounds (I-4a) include, for example, a methyl group substituted with a ^biphenyl group (that is, a biphenylmethyl group), a fluorine atom or a phenyl group substituted with a 4-methylpiperazin-1-ylsulfonyl group. a pyrrolidinyl group substituted with a benzyl group or a 5-indolylmethyl group, or the group (III) (wherein Y ³¹ is a methylene group and Q ³¹ is —NH— or —NH—C (=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, Q ³¹ is -NH-C(=O)-, and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-4a) include, for example, compounds represented by the following formulas (I-4a-1) to (I-4a-5) (hereinafter, “compound (I-4a-1)” etc.) and the like. These compounds are merely examples of preferred compounds (I-4a), and preferred compounds (I-4a) are not limited to these.

Preferred compounds (I-4b) include, for example, a phenyl group in which R ¹⁴² is substituted with a fluorine atom or a 4-methylpiperazin-1-ylsulfonyl group, or group (III) (in group (III), Y ³¹ is a methylene group, Q ³¹ is -NH- or -NH-C(=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, and Q ³¹ is -NH-C(=O)- and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-4b) include, for example, compounds represented by the following formulas (I-4b-1) to (I-4b-3) (hereinafter, “compound (I-4b-1)” etc.) and the like. These compounds are merely examples of preferred compounds (I-4b), and preferred compounds (I-4b) are not limited to these.

Preferred compounds (I-5a) include, for example, a methyl group substituted with a biphenyl group (that is, a biphenylmethyl group), a fluorine atom or a phenyl substituted with a 4-methylpiperazin-1-ylsulfonyl group at R ¹⁵¹ a pyrrolidinyl group substituted with a benzyl group or a 5-indolylmethyl group, or the group (III) (wherein Y ³¹ is a methylene group and Q ³¹ is —NH— or —NH—C (=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, Q ³¹ is -NH-C(=O)-, and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-5a) include, for example, compounds represented by the following formulas (I-5a-1) to (I-5a-5) (hereinafter, “compound (I-5a-1)” etc.) and the like. These compounds are merely examples of preferable compound (I-5a), and preferable compound (I-5a) is not limited to these.

Preferred compounds (I-5b) include, for example, a phenyl group in which R ¹⁵² is substituted with a fluorine atom or a 4-methylpiperazin-1-ylsulfonyl group, or group (III) (in group (III), Y ³¹ is a methylene group, Q ³¹ is -NH- or -NH-C(=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, and Q ³¹ is -NH-C(=O)- and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-5b) include, for example, compounds represented by the following formulas (I-5b-1) to (I-5b-3) (hereinafter, “compound (I-5b-1)” etc.) and the like. These compounds are merely examples of preferred compounds (I-5b), and preferred compounds (I-5b) are not limited to these.

Preferred compounds (I-6a) include, for example, a methyl group substituted with a biphenyl group (that is, a biphenylmethyl group), a fluorine atom or a phenyl substituted with a 4-methylpiperazin-1-ylsulfonyl group in R ¹⁶¹ a pyrrolidinyl group substituted with a benzyl group or a 5-indolylmethyl group, or the group (III) (wherein Y ³¹ is a methylene group and Q ³¹ is —NH— or —NH—C (=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, Q ³¹ is -NH-C(=O)-, and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-6a) include, for example, compounds represented by the following formulas (I-6a-1) to (I-6a-5) (hereinafter, “compound (I-6a-1)” etc.) and the like. These compounds are merely examples of preferred compounds (I-6a), and preferred compounds (I-6a) are not limited to these.

Preferred compounds (I-6b) include, for example, a phenyl group in which R ¹⁶² is substituted with a fluorine atom or a 4-methylpiperazin-1-ylsulfonyl group, or group (III) (in group (III), Y ³¹ is a methylene group, Q ³¹ is -NH- or -NH-C(=O)-, and R ³¹ is an unsubstituted cyclohexyl group; Y ³¹ is a methylmethylene group, and Q ³¹ is -NH-C(=O)- and R ³¹ is a phenyl group substituted with a halogen atom).

Specifically, preferred compounds (I-6b) include, for example, compounds represented by the following formulas (I-6b-1) to (I-6b-3) (hereinafter, “compound (I-6b-1)” etc.) and the like. These compounds are merely examples of preferred compounds (I-6b), and preferred compounds (I-6b) are not limited to these.

Among the compounds (I) described above, compound (I-1-1) or compound (I-1-2) is preferable because it has particularly excellent inhibitory activity against 3CL protease, as shown in the examples below. (I-1-1) is more preferred.

The protease inhibitor of this embodiment may contain a pharmaceutically acceptable salt of compound (I) instead of compound (I). As used herein, the term "pharmaceutically acceptable" generally means to the extent that side effects are not caused when appropriately administered to a subject animal.

The salt is preferably a pharmaceutically acceptable acid addition salt or basic salt.

Acid addition salts include salts with inorganic acids such as hydrochloric acid, phosphoric acid, hydrobromic acid and sulfuric acid; acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, citric acid, malic acid, benzoic acid Acids, salts with organic acids such as methanesulfonic acid and benzenesulfonic acid are included.

Basic salts include salts with inorganic bases such as sodium hydroxide, potassium hydroxide, ammonium hydroxide and magnesium hydroxide; salts with organic bases such as caffeine, piperidine, trimethylamine and pyridine.

<<Method for producing compound (I)>>
Compound (I) can be prepared, for example, according to the type of R ¹¹ , R ¹² , R ¹³ and R ¹⁴ by known reactions to give 1-amino-2-carboxybenzene or 1-amino-3-carboxybenzene can be produced by introducing the functional group described above into. Specifically, it is as follows.

<Method for producing compound (I-1)>
Among compounds (I), compound (I-1) is, for example,
A compound represented by the following general formula (I-1a) (hereinafter sometimes abbreviated as "compound (I-1a)") is reacted with 1-amino-2-carboxybenzene to obtain the following general formula: A step of obtaining a compound represented by formula (I-1b) (hereinafter sometimes abbreviated as "compound (I-1b)") (hereinafter sometimes abbreviated as "compound (I-1b) production step") there is); and
Compound (I-1b) and a compound represented by the following general formula (I-1c) (hereinafter sometimes abbreviated as "compound (I-1c)") are reacted to give compound (I- a step of obtaining 1) (hereinafter sometimes abbreviated as “compound (I-1) production step A”);
(hereinafter sometimes referred to as “method A for producing compound (I-1)”).

In the reaction formula above, R ¹¹¹ is the same as above.

Alternatively, compound (I-1) is, for example,
Compound (I-1c) and 1-amino-2-methoxycarbonylbenzene are reacted to obtain a compound represented by the following general formula (I-1d) (hereinafter abbreviated as "compound (I-1d)" (hereinafter sometimes abbreviated as “compound (I-1d) manufacturing step”); and
a step of hydrolyzing the ester bond of compound (I-1d) to obtain compound (I-1e) (hereinafter sometimes abbreviated as “compound (I-1e) production step”);
A step of reacting compound (I-1e) with compound (I-1a) to obtain compound (I-1) (hereinafter sometimes abbreviated as “compound (I-1) production step B”) );
(hereinafter sometimes referred to as “method B for producing compound (I-1)”).

In the reaction formula above, R ¹¹¹ is the same as above.

Each step will be explained in detail below.

[Method A for producing compound (I-1)]
(Compound (I-1b) manufacturing process)
In the compound (I-1b) production step, compound (I-1a) is reacted with 1-amino-2-carboxybenzene to obtain compound (I-1b). The process for producing compound (I-1b) is a known amidation reaction.

(1) Compound (I-1a)
Compound (I-1a) (1H-Imidazole-4-methylcarboxamide) is a known compound, and commercially available products can be obtained and used.

In addition, instead of compound (I-1a), a compound represented by the following general formula (I-1a') (hereinafter sometimes abbreviated as "compound (I-1a')") can be used. . Compound (I-1a') is obtained by substituting the formyl group of compound (I-1a) with a methoxycarbonyl group, and replacing the active hydrogen group of the 1H-imidazol-4-yl group with a triphenylmethyl group (trityl group). protected.

Compound (I-1a') is a known compound, and commercially available products can be obtained and used.

When compound (I-1a') is used, the following reaction is carried out before or after compound (I-1) production step A described below, preferably after compound (I-1) production step A. By doing so, the trityl group can be deprotected and the methoxycarbonyl group can be converted to a formyl group.
1) a reaction in which a methoxycarbonyl group is reduced with a known reducing agent and substituted with a methylhydroxyl group;
2) reaction to activate the active hydrogen group of the 1H-imidazol-4-yl group by deprotecting the trityl group under acidic conditions;
3) A reaction in which a methyl hydroxyl group is oxidized with a known oxidizing agent to form a formyl group.

The above reactions 1) to 3) are usually carried out in this order.

Examples of the reducing agent used in the above reaction 1) include sodium borohydride (NaBH ₄ ) and the like. These compounds may be used alone or in combination of two or more. When using 2 or more types together, those combinations and ratios can be selected arbitrarily.

The amount of the reducing agent used can be 0.5 to 20 times the molar amount of the compound to be reduced.

In the reduction reaction in 1) above, the reaction temperature can be -20°C or higher and 50°C or lower.

Examples of acids used in the above reaction 2) include Bronsted acids such as p-toluenesulfonic acid, formic acid, acetic acid, trifluoroacetic acid and hydrochloric acid; SnCl ₂ , BiCl ₃ , CeCl ₃ , FeCl ₃ , BF ₃ Acids such as _ZnBr2 . These compounds may be used alone or in combination of two or more. When using 2 or more types together, those combinations and ratios can be selected arbitrarily.

The amount of acid used can be 0.001 to 20 times the molar amount of the trityl group in the compound.

In the above reaction 2), the reaction temperature is preferably -20°C or higher and 50°C or lower, more preferably -4°C or higher and 25°C or lower.

In the above reaction 2), the reaction time is preferably 1 minute or more and 30 hours or less, more preferably 1 minute or more and 20 hours or less.

Examples of the oxidizing agent used in the above reaction 3) include Dess-Martin periodinane (DMP).

The amount of the oxidizing agent used is preferably 0.5 to 2.0 times the molar amount of the compound to be oxidized, and 0.75 to 1.5 times the molar amount of the compound to be oxidized. and more preferably 0.8-fold molar amount or more and 1.2-fold molar amount or less.

In the oxidation reaction in 3) above, the reaction temperature is preferably -20°C or higher and 50°C or lower, more preferably -4°C or higher and 25°C or lower.

In the reaction of 3) above, the reaction time can be 10 minutes or more and 30 hours or less.

(2) Compound (I-1b)
Compound (I-1b) is a novel compound obtained in this step.

(3) Reaction Conditions The compound (I-1b) production process is a known amidation reaction.

In the compound (I-1b) production step, the reaction is preferably carried out using an amide condensing agent.

Examples of amide condensing agents include N,N'-dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDCl), 1-hydroxybenzotriazole (HOBT), 1-hydroxy -7-azabenzotriazole (HOAT), diphenyl phosphate azide (DPPA), O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) , 1-[bis(dimethylamino)methylene]-1H-benzotriazolium 3-oxide tetrafluoroborate (TBTU), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo( 4,5-b) pyridinium 3-oxide hexafluorophosphate (HATU), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo(4,5-b) pyridinium 3-oxide tetra fluoroborate (TATU) and the like. These compounds may be used alone or in combination of two or more. When using 2 or more types together, those combinations and ratios can be selected arbitrarily.

The amount of the amide condensing agent used can be, for example, 1.0 to 10.0 times the molar amount of 1-amino-2-carboxybenzene.

In the compound (I-1b) manufacturing process, it is preferable to carry out the reaction in the presence of a base.

Examples of the base include trialkylamines such as triethylamine and N,N-diisopropylethylamine (N,N-diisopropylethylamine; DIEA). These compounds may be used alone or in combination of two or more. When using 2 or more types together, those combinations and ratios can be selected arbitrarily.

The amount of base used can be, for example, 0.1 to 10 times the molar amount of 1-amino-2-carboxybenzene.

An aprotic solvent is preferably used as a reaction solvent in the compound (I-1b) manufacturing process.

The aprotic solvent is not particularly limited, but examples include pentane, hexane, cyclohexane, methylcyclohexane, decahydronaphthalene, toluene, triethylamine, tert-butyl methyl ether, chloroform, ethyl acetate, 1,2-dimethoxyethane, 2- Methoxyethyl ether, 1,2-dimethoxyethane, tetrahydrofuran, pyridine, 2-butanone, acetone, N-methylpyrrolidinone, nitromethane, acetonitrile, sulfolane, dimethylsulfoxide, diisopropylethylamine, isopropyl acetate, N,N-dimethylformamide, propylene carbonate etc.

The aprotic solvent may be used alone, or two or more of them may be used in combination. When two or more of them are used in combination, their combination and ratio can be arbitrarily selected.

The amount of the solvent used can be 10 to 1000 times the molar amount of 1-amino-2-carboxybenzene.

In the process for producing compound (I-1b), the amount of compound (I-1a) used is 0.5-fold molar amount or more and 2.0-fold molar amount or less than the amount of 1-amino-2-carboxybenzene used. is preferred, more preferably 0.75-fold molar amount or more and 1.5-fold molar amount or less, and even more preferably 0.8-fold molar amount or more and 1.2-fold molar amount or less.

In the manufacturing process of compound (I-1b), the reaction may be carried out under an inert gas atmosphere.

The inert gas is not particularly limited, but examples include nitrogen and argon.

The inert gas may be used alone, or two or more of them may be used in combination. When two or more of them are used, their combination and ratio can be arbitrarily selected.

In the compound (I-1b) manufacturing process, the reaction temperature can be -20°C or higher and 50°C or lower.

In the compound (I-1b) manufacturing process, the reaction time can be 10 minutes or more and 30 hours or less.

In the compound (I-1b) production process, after the reaction is completed, post-treatment may be performed as necessary by a known method to extract compound (I-1b). That is, post-treatment operations such as filtration, washing, extraction, pH adjustment, dehydration, concentration, etc., are carried out either singly or in combination of two or more as appropriate, and concentration, crystallization, reprecipitation, and column chromatography are performed. The compound (I-1b) can be taken out by, for example. Further, the extracted compound (I-1b) may be further subjected to any one of operations such as crystallization, reprecipitation, column chromatography, extraction, stirring and washing of crystals with a solvent, or a combination of two or more of them. You may refine|purify by carrying out one or more times.

In the step of producing compound (I-1b), after completion of the reaction, compound (I-1b) may be used in the next step without being taken out, but the yield of the target compound (I-1) is improved. Therefore, it is preferable to isolate compound (I-1b) by the method described above.

(Compound (I-1) production step A)
In compound (I-1) production step A, compound (I-1b) and compound (I-1c) are reacted to obtain compound (I-1). Compound (I-1) production step A is a known amidation reaction.

(1) Compound (I-1c)
R ¹¹¹ in compound (I-1c) is the same as R ¹¹¹ in compound (I-1) above.

Compound (I-1c) can also be obtained by oxidizing a primary alcohol or aldehyde having R ¹¹¹ with potassium chromate or the like.

(2) Reaction Conditions Various reaction conditions in the compound (I-1) production process A are the same as those in the compound (I-1b) production process.

In compound (I-1) production step A, the amount of compound (I-1c) used is 0.5-fold molar amount or more and 2.0-fold molar amount or less than the amount of compound (I-1b) used. It is preferably 0.75-fold molar amount or more and 1.5-fold molar amount or less, and further preferably 0.8-fold molar amount or more and 1.2-fold molar amount or less.

In compound (I-1) production step A, after completion of the reaction, compound (I-1) can be isolated in the same manner as in compound (I-1b) production step. 1) may be further purified by a similar method.

Each compound such as compound (I-1), compound (I-1a), compound (I-1b), and compound (I-1c) can be obtained by, for example, nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS) , infrared spectroscopy (IR), etc., can be used to confirm the structure.

[Method B for producing compound (I-1)]
In the production method B of compound (I-1), 1-amino-2-methoxycarbonylbenzene is used instead of 1-amino-2-carboxybenzene to hydrolyze the ester bond of compound (I-1d). , Production of the above compound (I-1) except that the compound (I-1e) converted to a carboxy group was obtained, and the order of reacting the compound (I-1a) and the compound (I-1c) was changed. A method similar to method A is used to obtain compound (I-1).

Specifically, in the method A for producing compound (I-1), the carboxy group of 1-amino-2-carboxybenzene is reacted with compound (I-1a) having R ¹³ and R ¹⁴ to obtain compound (I After obtaining -1b), the amino group of compound (I-1b) is reacted with compound (I-1c) having R ²¹ to obtain compound (I-1).

On the other hand, in the production method B of compound (I-1), the amino group of 1-amino-2-methoxycarbonylbenzene is reacted with compound (I-1c) having R ²¹ to obtain compound (I-1d). , After hydrolyzing the ester bond of compound (I-1d) to obtain compound (I-1e) converted to a carboxy group, a compound having R ¹³ and R ¹⁴ in the carboxy group of compound (I-1e) (I-1a) is reacted to obtain compound (I-1).

In the production process of compound (I-1d), by using 1-amino-2-methoxycarbonylbenzene instead of 1-amino-2-carboxybenzene, the carboxy group in 1-amino-2-carboxybenzene The yield of compound (I-1d) can be improved by suppressing the reaction between them or between the carboxy group of compound (I-1c) and 1-amino-2-carboxybenzene.

From the above, the compound (I-1d) manufacturing process has the same reaction conditions as the compound (I-1) manufacturing process A, and the compound (I-1) manufacturing process B is the same as the compound (I-1) manufacturing process. 1b) Since the manufacturing process and reaction conditions are the same, detailed description thereof is omitted.

(Compound (I-1e) manufacturing process)
In the compound (I-1e) production step, the ester bond of compound (I-1d) is hydrolyzed to obtain compound (I-1e). The compound (I-1e) production step is a known ester hydrolysis reaction.

(1) Compound (I-1d)
Compound (I-1d) is a novel compound obtained by an amidation reaction of 1-amino-2-methoxycarbonylbenzene and compound (I-1c). In compound (I-1d), R ¹¹¹ is the same as R ¹¹¹ in compound (I-1) above.

(2) Compound (I-1e)
Compound (I-1e) is a novel compound obtained in this step.

(3) Reaction Conditions In the step of producing compound (I-1e), the reaction is preferably carried out in the presence of a base.
Examples of the base include lithium hydroxide and the like.

The amount of the base used can be 1.0 to 10.0 times the molar amount of the compound (I-1d).

In the compound (I-1e) manufacturing process, the reaction temperature can be -20°C or higher and 50°C or lower.

In the compound (I-1e) manufacturing process, the reaction time can be 30 minutes or more and 10 hours or less.

<Method for producing compound (I-2)>
Among compounds (I), compound (I-2) is, for example,
The compound (I-1a) is reacted with 1-amino-3-carboxybenzene to obtain a compound represented by the following general formula (I-2a) (hereinafter abbreviated as "compound (I-2a)" (hereinafter sometimes abbreviated as “compound (I-2a) manufacturing step”); and
A step of reacting compound (I-2a) with compound (I-2b) to obtain compound (I-2) (hereinafter sometimes abbreviated as “compound (I-2) production step A” );
(hereinafter sometimes referred to as “method A for producing compound (I-2)”).

In the reaction formula above, R ¹²¹ is the same as above.

Alternatively, compound (I-2) is, for example,
Compound (I-2b) and 1-amino-3-methoxycarbonylbenzene are reacted to obtain a compound represented by the following general formula (I-2c) (hereinafter abbreviated as "compound (I-2c)" (hereinafter sometimes abbreviated as “compound (I-2c) manufacturing step”); and
A step of hydrolyzing the ester bond of compound (I-2c) to obtain a compound represented by the following general formula (I-2d) (hereinafter sometimes abbreviated as "compound (I-2d)") ( Hereinafter, it may be abbreviated as “compound (I-2d) manufacturing step”);
A step of reacting compound (I-2d) with compound (I-1a) to obtain compound (I-2) (hereinafter sometimes abbreviated as “compound (I-2) production step B”) );
It can be produced by a production method comprising

In the reaction formula above, R ¹²¹ is the same as above.

That is, as shown in the above reaction scheme, in the method A for producing compound (I-2), 1-amino-3-carboxybenzene is used instead of 1-amino-2-carboxybenzene for compound (I-1c). Instead of using compound (I-1b), in production method B of compound (I-2), 1-amino-3-methoxycarbonylbenzene is used instead of 1-amino-2-methoxycarbonylbenzene, compound (I- 1c) was replaced with compound (I-1b), using the same raw materials and the same method as in method A for producing compound (I-1) and method B for producing compound (I-1), respectively, to produce a compound (I-2) can be produced.

(Compound (I-2b))
Compound (I-2b) is a known carboxylic acid, and R ¹²¹ is the same as R ¹²¹ in compound (I-2) above.

Compound (I-2b) can also be obtained by oxidizing a primary alcohol or aldehyde having R ¹²¹ with potassium chromate or the like.

In compound (I-2) production method A and compound (I-2) production method B, after completion of the reaction, compound (I-2) is taken out in the same manner as in the compound (I-1b) production step. The compound (I-2) thus isolated may be further purified by a similar method.

Each compound such as compound (I-2), compound (I-2a), compound (I-2b), compound (I-2c), and compound (I-2d) can be obtained by, for example, nuclear magnetic resonance (NMR) spectroscopy , mass spectroscopy (MS), infrared spectroscopy (IR), and the like can be used to confirm the structure.

<Method for producing compound (1-3a) to compound (1-6b)>
Compound (1-3a) and compound (1-3b) described above, compound (1-4a) and compound (1-4b), compound (1-5a) and compound (1-5b), and compound (1- 6a) and compound (1-6b) are respectively a compound represented by the following formula (I-3c) (hereinafter sometimes abbreviated as "compound (I-3c)"), the following formula (I-4c ) (hereinafter sometimes abbreviated as "compound (I-4c)"), a compound represented by the following formula (I-5c) (hereinafter abbreviated as "compound (I-5c)" ), and a compound represented by the following formula (I-6c) (hereinafter sometimes abbreviated as “compound (I-6c)”) is used instead of the above compound (1-1a). can be manufactured by the manufacturing method described above. Compound (I-3c) can be produced by the method described in International Publication No. 2001-010894. Compound (I-4c) can be produced by the method described in WO2021-226546. Compound (I-5c) can be produced by the method described in WO2005-066123. Compound (I-6c) can be produced by the method described in WO2001-040189.

<<Antiviral pharmaceutical composition>>
The antiviral pharmaceutical composition of this embodiment contains the aforementioned protease inhibitor and a pharmaceutically acceptable carrier.

According to the antiviral pharmaceutical composition of the present embodiment, viral replication and proliferation can be effectively suppressed, and a therapeutic effect on viral infections is expected.

In addition, the antiviral pharmaceutical composition of the present embodiment can be preferably used particularly against infections caused by viruses having 3CL protease. Viruses that cause such infections include, for example, SARS-coronavirus-2, SARS-coronavirus, rhinovirus, Middle East respiratory syndrome coronavirus, and the like.

Next, the components contained in the antiviral pharmaceutical composition of this embodiment will be described in detail below. As the protease inhibitor, those exemplified in the above "protease inhibitor" can be used.

<Pharmaceutically acceptable carrier>
As the pharmaceutically acceptable carrier, those that are usually used for formulation of pharmaceutical compositions can be used without particular limitation. More specifically, for example, binders such as gelatin, corn starch, tragacanth gum, and gum arabic; excipients such as starch and crystalline cellulose; swelling agents such as alginic acid; solvents for injections such as water, ethanol, and glycerin; Adhesives such as rubber-based adhesives and silicone-based adhesives are included.

<Additive>
The antiviral pharmaceutical composition of this embodiment may contain additives. Additives include lubricants such as calcium stearate and magnesium stearate; sweeteners such as sucrose, lactose, saccharin, and maltitol; flavoring agents such as peppermint and red oil; stabilizers such as benzyl alcohol and phenol; buffers such as salts and sodium acetate; solubilizers such as benzyl benzoate and benzyl alcohol; antioxidants; preservatives and the like.

The antiviral pharmaceutical composition of the present embodiment is prepared by appropriately combining the protease inhibitor, the pharmaceutically acceptable carrier, and, if necessary, additives, and the generally accepted unit required for pharmaceutical practice. It can be formulated by blending in dosage forms.

The antiviral pharmaceutical composition of the present embodiment may be used in combination with at least one selected from the group consisting of therapeutic agents having antiviral activity other than the protease inhibitors and therapeutic agents for other diseases. . The protease inhibitor and other drug may be formulated in the same formulation or in separate formulations. In addition, each formulation may be administered via the same administration route or via separate administration routes. Furthermore, each formulation may be administered simultaneously, sequentially, or separately with a certain time or period of time between them. In one embodiment, the protease inhibitor and other drug may be provided as a kit containing them.

<Dosage form>
The antiviral pharmaceutical composition of this embodiment may be in a dosage form for oral use or may be in a dosage form for parenteral use. Dosage forms for oral use include, for example, tablets, capsules, elixirs, microcapsules and the like. Dosage forms for parenteral use include, for example, injections, ointments, patches and the like.

<Administration method>
Subjects to be administered include, but are not limited to, humans, monkeys, dogs, cows, horses, sheep, pigs, rabbits, mice, rats, guinea pigs, hamsters, and cells thereof. Among them, mammals or mammalian cells are preferred, and humans or human cells are particularly preferred.

Administration routes include, for example, intraarterial injection, intravenous injection, and subcutaneous injection; intranasal, transbronchial, intramuscular, transdermal, and oral administration; can do

The dosage of the antiviral pharmaceutical composition of this embodiment varies depending on the type of compound (I), the symptoms of the subject of administration, the site of administration, the method of administration, and the like. A person skilled in the art can appropriately select an appropriate dosage.

The administration of the antiviral pharmaceutical composition of this embodiment may be a single administration or multiple administrations. In the case of multiple administration, for example, every 2 hours or more and 12 hours or less, every day, or once every 2 days, 5 days, 1 week, 1.5 weeks, several weeks, 1 month or several months etc. can be administered.

<<Other Embodiments>>
In one embodiment, the present disclosure provides a method for preventing, treating, or suppressing progression of viral infections, comprising administering an effective amount of compound (I) to a patient or patient in need of treatment. do. Here, as the compound (I), those similar to those described above can be mentioned. In addition, the causative viruses of viral infections include those mentioned above.

In one embodiment, the present disclosure provides compound (I) for prevention, treatment, or inhibition of progression of viral infections. Here, as the compound (I), those similar to those described above can be mentioned. In addition, the causative viruses of viral infections include those mentioned above.

In one embodiment, the present disclosure provides use of compound (I) for manufacturing a protease inhibitor or antiviral pharmaceutical composition. Here, as the compound (I), those similar to those described above can be mentioned. In addition, the causative viruses of viral infections include those mentioned above.

Although the present disclosure will be described below with reference to examples, the present disclosure is not limited to the following examples.

In the following, for example, "compound represented by general formula (I)" is referred to as "compound (I)". .

[Production Example 1-1]
(Production of compound (I-1-5))
1. Production of Compound (13) Compound (13) was produced through the route shown below.

DIEA (28.3 g, 219 mmol, 38.0 mL) was added to a solution of compound (12) (30.0 g, 73.0 mmol) and compound (2) (10.0 g, 73.0 mmol) dissolved in DMF (100 mL) in advance. ), EDCI (21.0 g, 109 mmol) and HOBt (15 g, 109 mmol) were added. The mixture was stirred at 25° C. for 16 hours. The reaction mixture was diluted with EtOAc (400 mL), washed with water (100 mL x 3), dried over sodium sulfate, filtered, concentrated and concentrated under reduced pressure to give a residue. This was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=1/0→10/1 (volume ratio)) to obtain compound (13) (17.0 g, yield 43% by mass). Obtained as a yellow solid.

The measurement results of ¹ H-NMR and LCMS of compound (13) are shown below.
¹ H-NMR: (400 MHz, CDCl3) δ 8.07-8.13 (m, 1H), 7.48-7.53 (m, 1H), 7.39-7.42 (m, 1H), 7.30-7.34 (m, 9H), 7.16- 7.22 (m, 1H), 7.08-7.13 (m, 6H), 6.57-6.67 (m, 3H), 5.60 (br s, 1H), 5.27-5.31 (m, 1H), 4.87-4.96 (m, 1H) , 3.64 (s, 3H), 3.05-3.17 (m, 2H).

Then, compound (I-1-5) was produced from compound (13) by the route shown below.

2. Preparation of compound (29) HATU (1.43 g, 3 .77 mmol) and DIEA (730 mg, 5.65 mmol) were added. The mixture was stirred at 25° C. for 10 hours. The reaction mixture was extracted with EtOAc (50 mL), washed with water (30 mL x 3), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. This was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=1/0→10/1) to obtain compound (29) (1.03 g, yield 77% by mass).

The results of ¹ H-NMR measurement of compound (29) are shown below.
¹ H-NMR: (400 MHz, CD3OD) δ 8.56 (d, J = 8.4 Hz, 1H), 7.59-7.65 (m, 1H), 7.48-7.55 (m, 1H), 7.37-7.41 (m, 1H) , 7.24-7.31 (m, 9H), 7.05-7.14 (m, 7H), 6.72-6.76 (m, 1H), 4.89-4.90 (m, 1H), 3.91 (d, J = 1.2 Hz, 2H), 3.71 (s, 3H), 3.16-3.24 (m, 1H), 3.01-3.10 (m, 1H), 2.29-2.38 (m, 1H), 2.04 (s, 1H), 1.91-1.97 (m, 2H), 1.65 -1.79 (m, 3H), 1.21-1.44 (m, 5H).

3. Production of compound (I-1-5) Using the above compound (29) (101 mg, 0.14 mmol), "6. Production of compound (10) (alcoholization step)", "7 Production of compound (11) (detritylation step)” and “8. After converting the group to a hydroxymethyl group and deprotecting the trityl group, the hydroxymethyl group was converted to an aldehyde group to give compound (I-1-5) (12.9 mg, 0.030 mmol, yield 30% by mass) was obtained.

The results of ¹ H-NMR measurement of compound (I-1-5) are shown below.
¹ H-NMR (CD3OD, MeOH, δ (ppm)): 1.20-1.41 (3H, m), 1.41-1.51 (2H, m), 1.66-1.77 (1H, m), 1.77-1.88 (2H, m) , 1.95-2.04 (2H, m), 2.37 (1H, J = 11.5, 3.4 Hz, tt), 2.84-2.97 (1H, m), 3.05 (1H, J = 15.1, 9.6, 4.2 Hz, ddd), 3.94 (2H, s), 4.30-4.43 (1H, m), 4.66 (1H, J = 11.9, 4.2 Hz, dd), 6.84 (1H, J = 5.2 Hz, d), 7.09-7.19 (1H, m), 7.40-7.50 (1H, m), 7.56 (1H, s), 7.60 (1H, J = 7.8 Hz, d), 8.33-8.41 (1H, m).

[Production Example 1-2]
(Production of compound (I-1-1))
Compound (I-1-1) was produced as compound (I) by the route shown below. In the formulas below, Trt represents a trityl group.

1. Preparation of compound (3) 1-[bis(dimethylamino)methylene]-1H-1,2,3- triazolo(4,5-b)pyridinium 3-oxide hexafluorophosphate (HATU) (16.9 g, 44.4 mmol), N,N-diisopropylethylamine (DIEA) (12.0 g, 92.5 mmol), and compound (2) (1-amino-2-methoxycarbonylbenzene) (5.59 g, 37.0 mmol) was added and the mixture was stirred at 40° C. for 40 hours. The product was confirmed by thin layer chromatography (TLC) (petroleum/ethyl acetate (EtOAc) = 3/1 (v/v), Rf = 0.6, UV). The mixture was diluted with EtOAc (500 mL), washed with water (400 mL x 3) and brine (400 mL), dried over sodium sulfate, filtered and concentrated to give a yellow oil. This was purified by column chromatography (SiO ₂ , petroleum/EtOAc=1/0→3/1 (volume ratio)) to obtain compound (3) (8.80 g, yield 73% by mass).

The measurement results of liquid chromatography-mass spectrometry (LCMS) of compound (3) are shown below.
LCMS: m/z=267.0 (M-56+H) ⁺

2. Preparation of compound (4) To a pre-dissolved solution of compound (3) (5.00 g, 15.5 mmol) in CH ₂ Cl ₂ (25 mL) was added HCl/dioxane (4 M, 25 mL), then the mixture was heated to 20°C. and stirred for 1 hour. The mixture gradually turned into a white suspension. TLC (petroleum/ethyl acetate (EtOAc) = 3/1 (v/v), Rf = 0.05, UV) confirmed the completion of the reaction. The suspension was filtered and the filter cake was dried in vacuum to give compound (4) (4.00 g, 99% yield, HCl).

3. Preparation of compound (6) Compound (4) (4.00 g, 15.5 mmol, HCl) and compound (5) (2.17 g, 15.5 mmol) were previously dissolved in DMF (60 mL). Benzotriazole (HOBt) (3.13 g, 23.2 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI) (4.45 g, 23.2 mmol) and DIEA (7.99 g, 61.2 mmol). 8 mmol) was added. The mixture was then stirred at 20° C. for 16 hours. LCMS (EB3308-3-P1A) confirmed the MS signal of the product, m/z=345.0 (M+H) ⁺ . The mixture was diluted with EtOAc (300 mL), washed with water (200 mL x 3) and brine (200 mL), dried over sodium sulfate, filtered and concentrated to give a yellow solid. The solid was purified by trituration with petroleum/EtOAc (5/1 v/v, 50 mL) to give compound (6) (3.32 g, 62 wt% yield).

The results of ¹ H-NMR measurement of compound (6) are shown below.
¹ H-NMR: (400 MHz, CDCl ₃ ) δ 11.55 (br s, 1H), 8.68 (dd, J = 8.4 Hz, 0.8 Hz, 1H), 8.04 (dd, J = 8.0 Hz, 1.6 Hz, 1H) , 7.60-7.67 (m, 2H), 7.53-7.58 (m, 1H), 7.38-7.46 (m, 1H), 7.18-7.24 (m, 1H), 7.10-7.16 (m, 1H), 6.97 (br d , J = 6.8 Hz, 1H), 4.84-4.93 (m, 1H), 3.91 (s, 3H), 1.64 (d, J = 7.2 Hz, 3H).

4. Preparation of compound (7) Lithium hydroxide monohydrate (1.21 g, 28.9 mmol) and Water (20 mL) was added and the mixture was stirred at 20° C. for 1 hour. LCMS (EB3308-3-P1A) confirmed the MS signal of the product, m/z=331.0 (M+H) ⁺ . The mixture was concentrated to remove THF and the residue was adjusted to pH 4 with 1N HCl. Then extracted with EtOAc (50 mL x 2). After combining the two EtOAc phases, it was dried over sodium sulfate, filtered and concentrated to give compound (7) (3.18 g, 100 wt% yield).

The LCMS measurement results of compound (7) are shown below.
LCMS: m/z=331.0 (M+H) ⁺

5. Preparation of compound (9) HATU (5 .50 g, 14.5 mmol) and DIEA (3.74 g, 28.9 mmol) were added. The mixture was then stirred at 20° C. for 16 hours. It was confirmed by TLC (petroleum/EtOAc=1/1 (volume ratio), Rf=0.2, UV) that the raw material was consumed and the reaction product was formed. The mixture was diluted with EtOAc (300 mL), washed with water (200 mL x 3) and brine (200 mL), dried over sodium sulfate, filtered and concentrated to give a yellow solid. The solid was purified by column chromatography (SiO ₂ , petroleum/EtOAc=3/1→0/1 (v/v)) to give the product as a white solid. This white solid was subjected to prep-SFC (column: REGIS (S, S) WHELK-O1 (250 mm × 25 mm, 10 μm); mobile phase: [ethanol containing 0.1% by mass ammonium hydroxide]; B%: 50%-50 %, min) to give compound (9) (1.25 g, 17 wt % yield) as a white solid.

The results of ¹ H-NMR measurement of compound (9) are shown below.
¹ H-NMR: EB3308-5-P1HN (400 MHz, CDCl ₃ ) δ 8.55 (d, J = 8.4 Hz, 1H), 7.75 (d, J = 7.6 Hz, 1H), 7.69 (dt, J = 10.0 Hz , 2.0 Hz, 1H), 7.58 (dd, J = 8.0 Hz, 1.2 Hz, 1H), 7.49-7.56 (m, 2H), 7.42-7.48 (m, 1H), 7.23-7.32 (m, 10H), 7.10 -7.16 (m, 1H), 7.03-7.09 (m, 6H), 6.75 (s, 1H), 4.68-4.75 (m, 1H), 4.57 (q, J = 7.2Hz, 1H), 3.58 (s, 3H) ), 3.11 (dd, J = 14.8 Hz, 4.8 Hz, 1H), 2.95 (dd, J = 14.8 Hz, 9.6 Hz, 1H), 1.53 (d, J = 7.2 Hz, 3H).

6. Preparation of compound (I-1-1) Compound (I-1-1) was obtained from compound (9) in the same manner as in the preparation of compound (I-1-5) from compound (29).

The results of ¹ H-NMR measurement of compound (I-1-1) are shown below.
1H-NMR (CD3OD, MeOH, δ (ppm)): 1.56 (3H, J = 7.3 Hz, d), 2.75-3.05 (2H, m), 4.17-4.33 (1H, m), 4.51-4.71 (2H, m), 6.81 (1H, J = 6.5 Hz, 1H), 7.12 (1H, J = 7.6, 1.0 Hz, td), 7.24-7.36 (1H, m), 7.40-7.64 (4H, m), 7.70-7.86 (2H, m), 8.32-8.46 (1H, m).

[Production Example 1-3]
(Production of compound (I-1-2))
Compound (I-1-2) was produced as compound (I) by the route shown below.

1. Preparation of compound (15) HATU (2.87 g) was dissolved in DMF (15 mL) in advance with compound (13) (2.00 g, 3.77 mmol) and compound (14) (1.60 g, 7.54 mmol). , 7.54 mmol) and DIEA (1.46 g, 11.3 mmol) were added. The mixture was stirred at 25° C. for 10 hours. The reaction mixture was diluted with EtOAc (75 mL), washed with water (30 mL x 3), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. This was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=10/0→10/1 (volume ratio)) to give compound (15) (2.07 g, yield 75.0% by mass). got

The results of ¹ H-NMR measurement of compound (15) are shown below.
¹ H-NMR: (400 MHz, CD ₃ OD) δ 8.34 (d, J = 8.4 Hz, 1H), 7.54 (dd, J = 8.0 Hz, 1.2 Hz, 1H), 7.45-7.50 (m, 1H), 7.38 (d, J = 1.2 Hz, 1H), 7.23-7.31 (m, 17H), 7.17-7.20 (m, 1H), 7.08-7.12 (m, 1H), 7.03-7.07 (m, 6H), 6.68 (s, 1H), 4.77 (dd, J = 9.6Hz, 4.8Hz, 1H), 3.70 (s, 3H), 3.62 (s, 2H), 3.11-3.18 (m, 1H), 3.00 (dd, J = 14.8Hz, 9.6Hz, 1H).

3. Preparation of compound (I-1-2) Compound (I-1-2) was obtained from compound (15) in the same manner as in the preparation of compound (I-1-5) from compound (29).

The ¹ H-NMR measurement results of compound (I-1-2) are shown below.
¹ H-NMR (CD3OD, MeOH, δ (ppm)): 2.78-2.91 (1H, m), 3.00 (1H, J = 14.8, 11.2, 4.4 Hz, ddd), 3.67 (2H, s), 4.20-4.32 (1H, m), 4.55-4.62 (1H, m), 6.81 (1H, J = 5.7 Hz, d), 7.05-7.14 (1H, m), 7.20-7.32 (5H, m), 7.32-7.42 (5H , m), 7.46-7.54 (2H, m), 8.14 (1H, J = 18.7, 8.4 Hz, dd).

[Production Example 1-4]
(Production of compound (I-1-3))
Compound (I-1-3) was produced as compound (I) by the route shown below.

1. Preparation of compound (19) HATU (2.87 g) was dissolved in DMF (10 mL) in advance with compound (13) (2.00 g, 3.77 mmol) and compound (18) (1.55 g, 7.54 mmol). , 7.54 mmol) and DIEA (1.46 g, 11.3 mmol) were added. The mixture was stirred at 25° C. for 10 hours. The reaction mixture was diluted with EtOAc (75 mL), washed with water (30 mL x 3), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. This was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=10/0→10/1 (volume ratio)) to obtain compound (19) (1.90 g, 2.51 mmol).

The ¹ H-NMR measurement results of compound (19) are shown below.
¹ H-NMR: EB2953-14-P1Z2 (400 MHz, CD3OD) δ 8.38 (d, J = 7.6 Hz, 1H), 7.56 (dd, J = 7.6 Hz, 1.2 Hz, 1H), 7.43-7.48 (m, 1H), 7.37-7.41 (m, 3H), 7.22-7.29 (m, 9H), 7.09-7.18 (m, 4H), 7.03-7.06 (m, 6H), 6.75 (s, 1H), 4.89-4.92 ( m, 1H), 3.81 (d, J = 12.8 Hz, 1H), 3.69 (s, 3H), 3.57 (d, J = 12.8 Hz, 1H), 3.17-3.23 (m, 2H), 3.05-3.12 (m , 2H), 2.37 (td, J = 9.2 Hz, 6.8 Hz, 1H), 2.14-2.25 (m, 1H), 1.71-1.84 (m, 3H).

2. Preparation of compound (I-1-3) Compound (I-1-3) was obtained from compound (19) in the same manner as in the preparation of compound (I-1-5) from compound (29).

The results of ¹ H-NMR measurement of compound (I-1-3) are shown below.
¹ H-NMR (CD3OD, MeOH, δ (ppm)): 1.71-1.95 (3H, m), 2.17-2.30 (1H, m), 2.39-2.51 (1H, m), 2.85-2.99 (1H, m) , 3.01-3.12 (1H, m), 3.15-3.25 (2H, m), 3.63-3.74 (1H, m), 3.76-3.87 (1H, m), 4.37-4.51 (1H, m), 4.62-4.68 ( 1H, m), 6.81-6.88 (1H, m), 7.05-7.31 (4H, m), 7.34-7.48 (3H, m), 7.48-7.58 (2H, m), 8.12-8.22 (1H, m).

[Production Example 1-5]
(Production of compound (I-1-4))
Compound (I-1-4) was produced as compound (I) by the route shown below.

1. Preparation of compound (23) Sodium hydrogen carbonate (317 mg, 3.77 mmol) and compound (22) ( 1.28 g, 11.3 mmol) was added. The mixture was then stirred at 25° C. for 1 hour. TLC (CH ₂ Cl ₂ /MeOH=10/1 (v/v), Rf=0.6) confirmed the completion of the reaction. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The EtOAc phases were combined, then dried over sodium sulfate, filtered and concentrated under reduced pressure to give compound (23) (2.19 g, 91 wt% yield).

The results of ¹ H-NMR measurement of compound (23) are shown below.
¹ H-NMR: (400 MHz, CDCl ₃ ) δ 11.99-12.03 (m, 1H), 8.54-8.61 (m, 2H), 7.69-7.74 (m, 1H), 7.40-7.47 (m, 2H), 7.26 -7.31 (m, 9H), 7.07-7.12 (m, 1H), 7.01-7.06 (m, 6H), 6.56 (s, 1H), 4.91-4.96 (m, 1H), 4.09-4.11 (m, 2H) , 3.57-3.60 (m, 3H), 3.04-3.18 (m, 2H).

2. Preparation of compound (25) Potassium carbonate (1.50 g, 10.82 mmol) and compound (24) (358 mg) were added to a pre-dissolved solution of compound (23) (2.19 g, 3.61 mmol) in acetonitrile (20 mL). , 3.61 mmol) was added. The mixture was stirred at 80° C. for 2 hours. It was confirmed by TLC (CH ₂ Cl ₂ /MeOH=10/1 (volume ratio), Rf=0.53) that the raw materials were completely consumed and the reaction product was produced. The mixture was diluted with water (20 mL) and extracted with 30 mL of EtOAc (10 mL x 3). After combining the EtOAc phases, they were dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. This was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=100/0→30/1 (volume ratio)) to obtain compound (25) (1.31 g, yield 52% by mass). rice field.

The results of ¹ H-NMR measurement of compound (25) are shown below.
¹ H-NMR: (400 MHz, CD ₃ OD) δ12.01 (br s, 1H), 8.54 (br d, J = 8.4 Hz, 1H), 7.71 (dd, J = 8.0 Hz, 1.2 Hz, 1H) , 7.44-7.50 (m, 1H), 7.42 (d, J = 1.2 Hz, 1H), 7.29-7.36 (m, 9H), 7.03-7.16 (m, 8H), 6.59 (s, 1H), 4.94 (dt , J = 7.6 Hz, 4.8 Hz, 1H), 3.69 (br d, J = 6.8 Hz, 2H), 3.63 (s, 3H), 3.06-3.20 (m, 2H), 2.63-2.81 (m, 1H), 1.98-2.11 (m, 2H), 1.55-1.87 (m, 6H), 1.20-1.45 (m, 5H).

3. Preparation of compound (I-1-4) Compound (I-1-4) was obtained from compound (25) in the same manner as in the preparation of compound (I-1-5) from compound (29).

The results of ¹ H-NMR measurement of compound (I-1-4) are shown below.
¹ H-NMR (CD3OD, MeOH, δ (ppm)): 1.10-1.35 (5H, m), 1.56-1.65 (1H, m), 1.70-1.79 (2H, m), 1.87-1.98 (2H, m) , 2.37-2.47 (1H, m), 2.85-2.96 (1H, m), 3.06 (1H, J = 15.1, 12.0, 4.5 Hz, ddd), 3.33-3.43 (2H, m), 4.33-4.43 (1H, m), 4.66 (1H, J = 9.6, 4.0 Hz, dd), 6.86 (1H, J = 3.5 Hz, d), 7.08-7.17 (1H, m), 7.40-7.47 (1H, m), 7.48-7.54 (1H, m), 7.56 (1H, J = 0.7 Hz, d), 8.32 (1H, J = 17.6, 8.1 Hz, dd).

[Production Example 1-6]
(Production of compound (I-1-6))
Compound (I-1-6) was produced as compound (I) by the route shown below.

1. Preparation of compound (33) EDCI (16.3 g , 84.8 mmol) was added. The mixture was stirred at 25° C. for 3 hours. The reaction mixture was diluted with EtOAc (150 mL), washed with water (300 mL x 2), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. This was purified by high-performance liquid column chromatography (C18, water (0.05 w/v% ammonium hydroxide + 10 mM ammonium carbonate)/acetonitrile = 45/55 → 95/5 (volume ratio)) to give compound (33) ( 1.15 g, 1.47 mmol).

The results of ¹ H-NMR measurement of compound (33) are shown below.
¹ H-NMR: δ 8.27-8.32 (m, 1H), 7.54-7.60 (m, 1H), 7.41-7.49 (m, 2H), 7.32-7.35 (m, 1H), 7.24-7.30 (m, 3H) , 7.16-7.23 (m, 7H), 7.08-7.15 (m, 3H), 6.97-7.03 (m, 6H), 6.71-6.75 (m, 1H), 6.31-6.35 (m, 1H), 4.96 (dd, J = 10.0 Hz, 4.4 Hz, 1H), 3.80-3.95 (m, 1H), 3.71 (s, 3H), 3.55-3.66 (m, 1H), 3.18-3.25 (m, 2H), 3.03-3.15 (m , 2H), 2.39-2.51 (m, 1H), 2.10-2.25 (m, 1H), 1.66-1.88 (m, 3H).

2. Preparation of compound (I-1-6) Compound (I-1-6) was obtained from compound (33) in the same manner as in the preparation of compound (I-1-5) from compound (29).

The results of ¹ H-NMR measurement of compound (I-1-6) are shown below.
¹ H-NMR (CD3OD, MeOH, δ (ppm)): 1.71-1.93 (3H, m), 2.13-2.31 (1H, m), 2.45-2.60 (1H, m), 2.88-3.00 (1H, m) , 3.00-3.12 (1H, m), 3.13-3.28 (2H, m), 3.73-4.00 (2H, m), 4.37-4.52 (1H, m), 4.60-4.68 (1H, m), 6.28-6.39 ( 1H, m), 6.78-6.90 (1H, m), 7.02-7.39 (4H, m), 7.42-7.59 (3H, m), 7.87-8.08 (1H, m), 8.08-8.25 (1H, m).

[Production Example 2-1]
(Production of compound (I-2-1))
Compound (I-2-1) was produced as compound (I) by the route shown below.

1. Preparation of compound (37) Compound (5) (5.00 g, 35.7 mmol) and compound (26) (1-amino-3-methoxycarbonylbenzene) (5.39 g, 35.7 mmol) were added to DMF (30 mL) in advance. was dissolved, HOBt (7.23 g, 53.5 mmol), EDCI (10.3 g, 53.5 mmol), DIEA (13.8 g, 107 mmol, 18.6 mL) were added at once at 25°C. The mixture was stirred at 25° C. for 4 hours. TLC (petroleum ether/EtOAc=10/1 (v/v), UV) confirmed complete consumption of starting material. The residue was poured into water (50 mL) and extracted with EtOAc (50 mL x 3). The EtOAc phase was combined then washed with brine (50 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give crude product. This was purified by column chromatography (SiO ₂ , petroleum ether/EtOAc=1/0→1/1) to give compound (37) (8.20 g, yield 84% by mass).

The results of ¹ H-NMR measurement of compound (37) are shown below.
¹ H-NMR: (400 MHz, CDCl ₃ ) δ 8.15-8.16 (m, 1H), 8.04-8.06 (m, 1H), 7.96 (s, 1H), 7.84-7.86 (m, 1H), 7.63-7.66 (m, 2H), 7.46-7.50 (m, 2H), 7.23-7.27 (m, 1H), 3.93 (s, 3H).

2. Preparation of compound (38) Lithium hydroxide monohydrate (3 .78 g, 90.0 mmol) was added. The mixture was stirred at 25° C. for 14 hours. TLC (petroleum ether/EtOAc=1/1 (v/v), UV) confirmed complete consumption of the starting compound (37). The reaction mixture was adjusted to pH 3 with HCl (1N), then extracted with ethyl acetate (150 mL x 3). The ethyl acetate phases were combined, then washed with brine (200 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to give compound (38) (6.90 g, 89 wt% yield). .

The results of ¹ H-NMR measurement of compound (38) are shown below.
¹ H-NMR: (400 MHz, DMSO-d ₆ ) δ 12.92-13.09 (m, 1H), 10.49 (s, 1H), 8.41 (s, 1H), 8.03-8.05 (m, 1H), 7.78-7.85 (m, 2H), 7.68-7.70 (m, 1H), 7.57-7.63 (m, 1H), 7.47-7.51 (m, 2H).

3. Preparation of compound (39) Compound (38) (3.00 g, 11.6 mmol), HATU (6.60 g, 17.4 mmol), and DIEA (4.49 g, 34.7 mmol, 6.49 g, 34.7 mmol) were prepared in advance in DMF (30 mL). 05 mL) was dissolved, compound (8) (5.18 g, 11.6 mmol, HCl) was added in one portion at 25° C. under nitrogen. The mixture was stirred at 25° C. for 4 hours. TLC (CH ₂ Cl ₂ /MeOH=10/1 (v/v), UV) confirmed the completion of the reaction. The residue was poured into water (50 mL) and extracted with ethyl acetate (50 mL x 3). The ethyl acetate phases were combined then washed with brine (50 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give a residue. The residue was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=1/0→10/1) to give compound (39) (1.02 g, yield 13% by weight) as a white solid. rice field.

The results of ¹ H-NMR measurement of compound (39) are shown below.
¹ H-NMR: (400 MHz, MeOD) δ 8.11-8.12 (m, 1H), 7.88-7.90 (m, 2H), 7.77-7.79 (m, 1H), 7.66-7.70 (m, 1H), 7.55- 7.57 (m, 2H), 7.47-7.50 (m, 1H), 7.42 (s, 1H), 7.35-7.39 (m, 1H), 7.24-7.32 (m, 10H), 7.08-7.10 (m, 6H), 6.80 (s, 1H), 4.92- 4.94 (m, 1H), 3.74 (s, 3H), 3.20-3.25 (m, 1H), 3.04-3.11 (m, 1H).

4. Preparation of compound (I-2-1) Compound (I-2-1) was obtained from compound (39) in the same manner as in the preparation of compound (I-1-5) from compound (29).

The results of ¹ H-NMR measurement of compound (I-2-1) are shown below.
¹ H-NMR (CD3OD, MeOH, δ (ppm)): 2.87-2.97 (1H, m), 3.07 (1H, J = 15.0, 10.1, 4.1 Hz, ddd), 4.34-4.44 (1H, m), 4.66 (1H, J =7.5, 4.1 Hz, dd), 6.83-6.90 (1H, m), 7.31-7.38 (1H, m), 7.44 (1H, J = 8.0 Hz, t), 7.50-7.59 (3H, m ), 7.67-7.73 (1H, m), 7.79 (1H, J = 7.8 Hz, d), 7.81-7.88 (1H, br m), 8.04 (1H, s).

[Production Example 2-2]
(Production of compound (I-2-2))
Compound (I-2-2) was produced as compound (I) by the route shown below.

1. Preparation of compound (42) Compound (1) (2.50 g, 13.2 mmol), HATU (7.55 g, 19.9 mmol), and DIEA (5.13 g, 39.7 mmol, 6.5 g, 19.9 mmol) were added in advance to DMF (20 mL). 91 mL) was dissolved, Compound (26) (1-amino-3-methoxycarbonylbenzene) (2.00 g, 13.2 mmol) was added in one portion at 25°C. The mixture was stirred at 25° C. for 4 hours. It was confirmed by TLC (CH ₂ Cl ₂ /MeOH=20/1 (volume ratio), UV) that the starting compound (1) was completely consumed. The residue was poured into water (30 mL) and extracted with EtOAc (50 mL x 3). The extracted EtOAc phases were combined then washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give crude product. This was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=1/0→10/1) to obtain compound (42) (3.30 g, yield 76% by mass).

The results of ¹ H-NMR measurement of compound (42) are shown below.
¹ H-NMR: (400 MHz, CD ₃ OD) δ 8.25 (s, 1H), 7.81 (d, J = 7.6 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.42 (t, J = 8.0 Hz, 1H), 4.18-4.23 (m, 1H), 3.90 (s, 3H), 1.45 (s, 9H), 1.38 (d, J = 7.2 Hz, 3H).

2. Preparation of compound (43) Compound (42) (3.30 g, 10.2 mmol) and HCl/ _dioxane (4 M, 15 mL) were mixed in _CH2Cl2 (15 mL) at 25°C. The mixture was stirred at 25° C. for 1 hour. LCMS (EB2954-19-P1A) confirmed complete consumption of compound (42). The reaction mixture was concentrated in vacuo to give compound (43) (2.2 g, crude).

The LCMS measurement results of compound (31) are shown below.
LCMS: m/z=223.2 (M+H) ⁺

3. Preparation of compound (44) Compound (5) (1.39 g, 9.90 mmol), HOBt (2.01 g, 14.8 mmol), DIEA (3.84 g, 29.7 mmol, 5.17 mL) was prepared in advance in DMF (30 mL). ), and EDCI (2.85 g, 14.8 mmol), compound (43) (2.20 g, 9.90 mmol) was added in one portion at 25° C. under nitrogen. The mixture was stirred at 25° C. for 2 hours. TLC (CH ₂ Cl ₂ /MeOH=20/1 (v/v), UV) confirmed the completion of the reaction. The residue was poured into water (30 mL) and extracted with EtOAc (20 mL x 3). The extracted EtOAc phases were combined then washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give crude product. The crude product was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=1/0→10/1) to give compound (44) (3.20 g, yield 93% by weight).

The LCMS measurement results of compound (44) are shown below.
LCMS: m/z=345.1(M+H) ⁺ , R _t =0.519 min

4. Preparation of compound (45) To a solution of compound (44) (3.20 g, 9.29 mmol) previously dissolved in THF (21 mL) and water (7 mL) was added lithium hydroxide monohydrate (1.17 g, 27.5 mmol). 9 mmol) was added in one portion at 25° C. under nitrogen. The mixture was stirred at 25° C. for 12 hours. TLC (petroleum ether/EtOAc=1/1 (v/v), UV) confirmed the completion of the reaction. The reaction mixture was adjusted to pH 3 with HCl (1M) and extracted with EtOAc (10 mL x 3). The extracted EtOAc phases were combined then washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give compound (45) (2.20 g, 72 wt% yield). rice field.

The measurement results of ¹ H-NMR and LCMS of compound (45) are shown below.
¹ H-NMR: (400 MHz, CD ₃ OD) δ 8.25-8.26 (m, 1H), 7.85-7.87 (m, 1H), 7.72-7.78 (m, 2H), 7.63-7.66 (m, 1H), 7.41-7.53 (m, 2H), 7.28-7.32 (m, 1H), 4.66-4.71 (m, 1H), 1.56-1.57 (t, J = 4.0 Hz, 3H).

5. Preparation of compound (46) HATU (1.73 g, 4.54 mmol), DIEA (1.17 g, 9.80 mmol, 1.58 mL) and compound (45) (1.0 g, 3.03 mmol) were prepared in DMF (10 mL). ) was dissolved, compound (8) (1.25 g, 3.03 mmol) was added in one portion at 25°C under nitrogen. The mixture was stirred at 25° C. for 4 hours. TLC (petroleum ether/EtOAc=0/1 (v/v), UV) confirmed the completion of the reaction. The residue was poured into water (10 mL) and extracted with EtOAc (20 mL x 3). The extracted EtOAc phases were combined then washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether/EtOAc=1/0→0/1 (volume ratio)) to give compound (46) (1.00 g, yield 45% by mass). rice field.

The results of ¹ H-NMR measurement of compound (46) are shown below.
¹ H-NMR: (400 MHz, CD ₃ OD) δ 8.05-8.08 (m, 2H), 7.73-7.78 (m, 2H), 7.64-7.66 (m, 1H), 7.43-7.54 (m, 3H), 7.28-7.36 (m, 10H), 7.08-7.10 (m, 6H), 6.97 (s, 1H), 4.98-5.01 (m, 1H), 4.68-4.73 (m, 1H), 3.76 (s, 3H), 3.29-3.31 (m, 1H), 3.09-3.15 (m, 1H), 1.58 (t, J = 7.2 Hz, 3H).

6. Preparation of compound (I-2-2) Compound (I-2-2) was obtained from compound (46) in the same manner as in the preparation of compound (I-1-5) from compound (29).

The ¹ H-NMR measurement results of compound (I-2-2) are shown below.
¹ H-NMR (CD3OD, MeOH, δ (ppm)): 1.56 (3H, J = 7.1 Hz, d), 2.81-2.97 (1H, m), 2.97-3.14 (1H, m), 4.28-4.45 (1H , m), 4.57-4.74 (2H, m), 6.84 (1H, s), 7.23-7.42 (2H, m), 7.42-7.58 (3H, m), 7.58-7.80 (3H, m), 7.93 (1H , s).

[Production Example 2-3]
(Production of compound (I-2-3))
Compound (I-2-3) was produced as compound (I) by the route shown below.

1. Preparation of compound (51) To a mixture of compound (50) (3.09 g, 30.8 mmol) and triethylamine (TEA) (4.29 mL) in CH ₂ Cl ₂ (60 mL) was added in CH ₂ Cl ₂ (60 mL). A pre-dissolved solution of compound (49) (6.80 g, 30.8 mmol) was added dropwise at 0°C. After the addition, the yellow mixture was stirred at 25° C. for 1 hour under a nitrogen atmosphere. TLC (CH ₂ Cl ₂ /MeOH=10/1 (v/v), I ₂ ) confirmed the complete consumption of compound (49) with one major new spot of greater polarity as the product. Detected. The reaction mixture was concentrated to give a solid. The solid was triturated with CH ₂ Cl ₂ /MeOH (10/1 (v/v), 25 mL) at 25° C. for 10 min to give compound (51) (7.10 g, crude).

The results of ¹ H-NMR measurement of compound (51) are shown below.
¹ H-NMR: (400 MHz, DMSO-d ₆ ) δ 8.16 (d, J = 8.4 Hz, 2H), 7.84 (d, J = 8.4 Hz, 2H), 2.92 (br s, 4H), 2.33-2.37 (m, 4H), 2.14 (s, 3H).

2. Preparation of compound (52) HOBt (5.06 g, 37.4 mmol), EDCI (7.18 g, 37.9 mmol) and EDCI (7.18 g, 37.9 mmol) were added to a solution of compound (51) (7.10 g, 24.9 mmol) in DMF (7 mL). 4 mmol), DIEA (9.68 g, 74.9 mmol) and compound (36) (3.77 g, 24.99 mmol) were added. The yellow mixture was stirred at 25° C. for 4 hours under a nitrogen atmosphere. TLC (CH ₂ Cl ₂ /MeOH=20/1 (v/v), I ₂ ) confirmed the complete consumption of compound (51) with one major new spot of greater polarity as the product. Detected. The reaction mixture was diluted with water (200 mL) and then extracted with EtOAc (150 mL x 4). After combining the extracted EtOAc phases, they were dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. This was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=100/1→20/1) to give compound (52) (4.79 g, crude product) as a yellow oil.

The LCMS measurement results of compound (52) are shown below.
LCMS: m/z=418.3(M+H) ⁺ , R _t =0.433 min

3. Preparation of compound (53) Lithium hydroxide monohydrate (1.44 g, 34.4 mmol) and Water (1.6 mL) was added. The yellow mixture was stirred at 25° C. for 7 hours under a nitrogen atmosphere. TLC (CH ₂ Cl ₂ /MeOH=20/1 (v/v), I ₂ ) confirmed the complete consumption of compound (52) with one major new spot of greater polarity as the product. Detected. The reaction mixture was adjusted to p2 with 1N HCl aqueous solution and some precipitate formed. The precipitate was collected by filtration and dried in vacuo to give compound (53) (2.97 g, crude) as a yellow solid.

The measurement results of ¹ H-NMR and LCMS of compound (53) are shown below.
¹ H NMR: (400 MHz, CD ₃ OD) δ 8.24 (s, 1H), 8.07 (d, J = 8.4 Hz, 2H), 7.90 (d, J = 8.8 Hz, 1H), 7.82 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.61 (s, 1H), 7.38 (t, J = 7.6 Hz, 1H), 3.07 (br s, 4H), 2.62 (br s, 4H), 2.33 (s, 3H).

4. Preparation of compound (54) DIEA (2.54 g, 19.6 mmol, 3.42 mL) and HATU (3. 73 g, 9.82 mmol) was added. The reaction mixture was stirred for 30 minutes, then compound (8) (2.69 g, 6.54 mmol) was added. The mixture was stirred at 25° C. for 3 hours under a nitrogen atmosphere. TLC (CH ₂ Cl ₂ /MeOH=10/1 (v/v), I ₂ ) confirmed the complete consumption of compound (53) with one major new spot of greater polarity as the product. Detected. The reaction mixture was diluted with water (50 mL) and then extracted with EtOAc (50 mL x 3). The extracted EtOAc phases were combined, then washed with brine (50 mL x 2), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. This was purified by column chromatography (SiO ₂ , CH ₂ Cl ₂ /MeOH=100/1→20/1) to obtain compound (54) (1.05 g, yield 18.3 mass %).

The measurement results of ¹ H-NMR, LCMS, HPLC, and SFC of compound (54) are shown below.
¹ H-NMR: (400 MHz, CD ₃ OD) δ 8.14-8.16 (m, 3H), 7.95-7.97 (m, 2H), 7.88-7.90 (m, 1H), 7.61 (s, 1H), 7.55- 7.57 (m, 1H), 7.48-7.51 (m, 1H), 7.25-7.32 (m, 9H), 7.08 -7.09 (m, 6H), 6.84 (s, 1H), 4.93-4.95 (m, 1H), 3.73 (s, 3H), 3.30-3.32 (m, 1H), 3.22-3.27 (m, 4H), 3.08-3.12 (m, 1H), 3.04-3.05 (m, 4H), 2.63-2.65 (s, 3H) ).

5. Preparation of compound (I-2-3) Compound (I-2-3) was obtained from compound (54) in the same manner as in the preparation of compound (I-1-5) from compound (29).

The results of ¹ H-NMR measurement of compound (I-2-3) are shown below.
¹ H-NMR (CD3OD, MeOH, δ (ppm)): 2.27 (3H, s), 2.52 (4H, J = 4.8 Hz, t), 2.92 (1H, J = 14.9, 9.9, 5.4 Hz, ddd), 2.99-3.16 (5H, m), 4.40 (1H, J = 9.3, 9.3, 4.2 Hz, ddd), 4.61-4.70 (1H, m), 6.87 (1H, s), 7.40-7.50 (1H, m), 7.50-7.54 (1H, m), 7.54-7.58 (1H, s), 7.82-7.89 (1H, m), 7.89-7.92 (1H, m), 7.92-7.96 (1H, m), 8.05-8.11 (1H , m), 8.13-8.16 (1H, m), 8.16-8.20 (1H, m).

[Example 1]
(Inhibitory activity confirmation test for 3CL protease of SARS-CoV-2)
1. Preparation of SARS-CoV-2 3CL protease Expression of the SARS-CoV-2 3CL protease (R188I mutant) using E. coli has resulted in SARS-CoV-2 that is resistant to degradation and expresses stable catalytic activity. 3CL protease, a mutant in which the arginine residue at position 188 was replaced with an isoleucine residue was constructed and used by inverse PCR.

Specifically, glutathione-S-transferase (GST), His tag (His6), Flag tag, and SARS-CoV-2 3CL protease (R188I mutant, amino acid sequence shown in SEQ ID NO: 1) bind in this order. A pGEX-6P-1-based plasmid pGEX-6P-1/SARS-3CL-COV2 into which a base sequence (SEQ ID NO: 2) encoding the fusion protein was introduced was constructed and transfected into Escherichia coli DHα strain.

4 μg of the prepared expression vector pGEX-6P-1/SARS-3CL-COV2 was dissolved in 20 μL of sterile water (concentration: 200 ng/μL). Next, while cooling with ice, 1 μL (200 ng) of the dissolved expression vector was added to a 1.5 mL Eppendorf tube, and 50 μL of competent cells BL21Gold (DE3) were added, slowly stirred, and allowed to stand for 10 minutes. Thereafter, transformation was performed by a heat shock method using a water bath (42°C: 1 minute, once: 5 minutes). Further, 150 μL of SOC medium was added, and the mixture was slowly stirred and then incubated in a clean oven at 37° C. for 30 minutes. After incubation, 50 μL of the above solution was dropped onto the surface of an agar medium (LB-Ampicillin, Ampicillin concentration: 50 μg/mL) and uniformly coated with a Conlarge stick. After that, incubation was performed overnight at 37° C. in a constant temperature bath. After incubation, colonies grown on the agar medium were used to confirm the success or failure of introduction of the target gene by colony PCR. At this time, PGEX-5 and PGEX-3 were used as the forward and reverse sequencing primers, respectively. PCR products prepared from multiple colonies were subjected to gel electrophoresis and their bands were localized. A 15 v/v % glycerol solution was prepared from the LB-Ampicillin culture solution of the colonies in which introduction of the target gene was confirmed as described above, and stored at -80°C.

The aforementioned 15v/v% glycerol solution (prepared from the culture solution of colonies in which introduction of the target gene was confirmed) was inoculated on an agar medium (LB-Ampicillin, Ampicillin concentration is 50 µg/mL) and placed in a constant temperature bath. Incubation was carried out at 37°C overnight. After that, add 100 mL of LB medium (LB-Ampicillin, Ampicillin concentration is 50 μg/mL) to a 300 mL sterilized Erlenmeyer flask, and inoculate one colony grown on the agar medium to inoculate the LB in the Erlenmeyer flask. added to the medium. This culture solution was placed in a shaking incubator and cultured at 37° C. and a shaking speed of 180 rpm for 18 hours (preculture). Next, 1 L each of LB-Ampicillin medium (Ampicillin concentration: 50 μg/mL) was prepared in six 3-L Erlenmeyer flasks. After that, 10 mL each of the pre-culture solution was added to six 3-L Erlenmeyer flasks to carry out main culture. The culture temperature during the main culture was 37° C., and the shaking speed was 180 rpm. After 2.5 hours from the start of the main culture, when the OD600 (absorbance at a wavelength of 600 nm) reached 0.8, the culture was interrupted and the temperature was set to 28° C. and allowed to stand for 30 minutes. Thereafter, 0.5 mL of 1 M IPTG was added to each culture solution to make the final IPTG concentration 0.5 mM, and the culture was restarted at 28° C. and continued for 5 hours. After the main culture was completed, the culture solution was taken out and the cells were harvested using a centrifuge. The collected cells were washed twice with 1×PBS, frozen in liquid nitrogen and stored at -80°C. The yield of cells after culture was about 5 g per 1 L of LB-Ampicillin medium.

The cryopreserved E. coli cells were thawed on ice, and then 10 mL of Lysis Buffer and a reversible protease inhibitor per 1 g of the cells were added and suspended by adding 1/1000 volume of the Lysis Buffer. The composition of Lysis Buffer is 50 mM Tris-HCl pH 7.8, 250 mM NaCl, 1 mM DTT, and 10 mM Imidazole. The suspended cells were crushed by an ultrasonicator, and then centrifuged by an ultracentrifuge at 45,000 rpm for 45 minutes at 4°C to collect the supernatant fraction. Furthermore, the supernatant fraction was filtered with a disk filter with a pore size of 0.22 μm, and subjected to subsequent purification.

A Ni affinity column (CV = 5 mL) was equilibrated with a Ni affinity column equilibration buffer. The composition of the Equilibration Buffer is identical to that of the Lysis Buffer. After that, the supernatant (50 mL) after crushing was applied to the Ni column at a flow rate of 1 mL/min to collect a flow through (FT). The column was then washed with the same equilibration buffer (100 mL) (Wash fraction). After washing the Ni column, elution was performed with a Ni affinity column elution buffer. The composition of the elution buffer was 50 mM Tris-HCl pH 7.8, 250 mM NaCl, 1 mM DTT, and 500 mM Imidazole, and elution was performed in Linear Gradient mode with Imidazole concentrations ranging from 10 mM to 500 mM, and fractions were collected by 1 mL each. After the target protein was eluted, the column was washed with 50 mL of the same elution buffer. The collected eluted fractions were confirmed for bands by electrophoresis (SDS-PAGE), and a total volume of about 80 mL was collected as fractions containing the target protein. This collected fraction was concentrated to about 10 mL under centrifugation conditions of 3500 rpm and 4° C. using a centrifugal separation type membrane device.

The elution fraction concentrated to about 10 mL was cleaved with AcTEV Protease in order to cleave and remove GST and 6xHis-Tag. 50 μL of commercially available AcTEV Protease was added to the concentrated elution fraction, put into a dialysis tube, and cleaved with 1 L of Cleavage Buffer for 15 hours while dialysis was performed at 4°C. The composition of the Cleavage Buffer is 50 mM Tris-HCl pH 7.5, 200 mM NaCl, and 2 mM DTT. The solution after cutting was centrifuged (10,000 rpm, 60 minutes, 4° C.) and collected as a supernatant fraction.

Passive Ni affinity column chromatography was performed in order to completely remove uncleaved protein and GST and 6×His-Tag fragments after cleavage from the supernatant sample after removal of GST and 6×His-Tag. . First, a Ni affinity column (CV=5 mL) was equilibrated with Ni affinity column equilibration buffer. Next, the supernatant from which GST and 6×His-Tag had been cleaved and removed was applied to the Ni column at a flow rate of 1 mL/min to collect the FT fraction. Thereafter, the same Ni affinity column equilibrated buffer was applied to the column at a flow rate of 1 mL/min, and a total of 20 wash fractions (40 mL) were collected at a volume of 2 mL per Fraction. Finally, the Ni affinity column was washed with 50 mL of elution buffer containing 500 mM Imidazole. Each fraction was subjected to electrophoresis (SDS-PAGE) to confirm the presence or absence of the target protein and its purity.

In order to improve the purity of the collected target protein solution, anion exchange column chromatography was performed. First, dialysis was performed to reduce the NaCl concentration contained in the buffer of the protein solution. The composition of dialysis buffer is 20 mM Tris-HCl pH 8.0, 20 mM NaCl, and 1 mM DTT. The protein solution was placed in a dialysis tube, and dialysis was performed at a buffer volume of 1 L for 15 hours at 4°C. The protein solution after buffer replacement was purified using an anion exchange column (Hi TrapQ FF, CV=5 mL). First, the anion exchange column was equilibrated with an equilibration buffer. The composition of the Equilibration Buffer is the same as the Dialysis Buffer. After equilibrating the anion exchange column, the protein solution after buffer substitution was applied to the column at a flow rate of 1 mL/min to collect the FT fraction. After that, elution was sequentially performed with each elution buffer containing 100 mM NaCl, 200 mM NaCl, and 400 mM NaCl at a flow rate of 1 mL/min. The volume of each Fraction was 2 mL, and the total volume of elution was 30 mL. Each eluted fraction was confirmed by electrophoresis (SDS-PAGE). Furthermore, only the eluted fraction containing the target protein was collected, and using a centrifugation-type membrane device, concentration and buffer replacement were simultaneously performed under centrifugation conditions of 3500 rpm and 4°C. The buffer composition (final composition) for buffer replacement in this case was 20 mM Tris-HCl pH 7.8, 150 mM NaCl, 1 mM EDTA, and 1 mM DTT. The mass of the finally obtained protein was 5.5 mg. The yield was 1.1 mg per 1 g of cells.

2. Preparation of substrate peptide The substrate peptide was obtained from Reference 1 (Akaji K et al., “Evaluation of peptide-aldehyde inhibitors using R188I mutant of SARS 3CL protease as a proteolysis-resistant mutant.”, Bioorganic & Medicinal Chemistry, Vol. 16, Issue 21, pp. 9400-9408, 2008.) was used as the substrate peptide. Specifically, an undecapeptide substrate containing the P1/P2 cleavage sites of 3CL protease represented by the following amino acid sequence.

Substrate peptide: H-Thr-Ser-Ala-Val-Leu-Gln-Ser-Gly-Phe-Arg-Lys- _NH2 (SEQ ID NO: 3)

3. Inhibitory activity confirmation test Water (13.8 μL), 0.5 M Tris buffer (pH 7.5, 1 μL), 0.1 M dithiothreitol aqueous solution (1.7 μL), substrate peptide (0.8387 mM, 6.0 μL), SARS -In a mixture of CoV-2 3CL protease (R188I mutant) (14.7 μM, 1.5 μL), a compound prepared in each production example as an inhibitor (4 specimens of 25 μM or more and 1600 μM or less each, 1 μL each) Or DMSO (1 μL) was added and incubated at 37° C. for 1 hour. After adding 75 μL of 0.1% by mass trifluoroacetic acid aqueous solution to this reaction solution to make 100 μL, 85 μL of it was subjected to high performance liquid chromatography (elution conditions: acetonitrile containing 0.1% by mass trifluoroacetic acid and 0.1% by mass trifluoroacetic acid Gradient of acetic acid aqueous solution, acetonitrile concentration 10% by mass→20% by mass, 30 minutes). The cleavage rate at each inhibitor concentration calculated by comparing the area of the substrate peptide peak (elution time around 19.7 minutes) in the presence of DMSO with the substrate peptide area in the presence of the inhibitor as 100% cleavage rate. , the inhibition rate was determined. The 50% inhibition rate IC50 of the inhibitor was obtained from the sigmoid curve obtained by plotting the inhibition rate at each concentration. A similar test was performed using Rupintrivir (hereinafter sometimes referred to as "control compound (X)") as a control compound.

Compounds (I-1-1) to (I-1-6) and (I-2-1) to (I-2-3) have an IC50 of 1600 μM or less, especially compound (I-1-1) and (I-1-2) had a low IC50 of less than 500 μM, confirming particularly excellent inhibitory activity.

On the other hand, the control compound (X) could not satisfy the inhibition rate of 50% even at 1600 μM.

[Example 2]
(Docking simulation test for 3CL protease of SARS-CoV-2)
Co-crystal files of SARS-CoV-2 3CL protease and ligand were obtained from the Protein Data Bank, and a PDBQT file was created using AutoDock Tools, leaving those protein portions and removing the atoms of CYS145SG. Among the compounds shown below, each compound (I-3a-1) ~ (I-3a-5), (I-4a-1) ~ (I-4a-5), (I-5a-1), ( I-5a-2), (I-5a-5), (1-6a-1), (1-6a-5), (I-3b-1) ~ (I-3b-3), (I- 4b-1) to (I-4b-3), (1-5b-3), and (I-6b-1) protonated the aminic nitrogen atoms and their conformations were determined using the EmbedMolecule method of the RDKit. generated.

A docking simulation between the obtained PDBQT file and each compound was performed using QVina2. Among poses in which a carbon atom exists and an N atom of a pyrrolidone group or an imidazole group exists within 5 Å of HIS163NE, the pose with the highest score was selected as the docking score. Based on the ligand, the more negative the numerical value, the smaller the binding free energy and the more excellent the inhibitory activity. On the other hand, if no such docking pose was found, the docking score was set to 0. The docking pose was similarly searched for the control compound (X) used in Example 1, and the docking score was calculated.

Compounds (I-3a-1) ~ (I-3a-5), (I-4a-1) ~ (I-4a-5), (I-5a-1), (I-5a-4), ( I-5a-5), (1-6a-1), (1-6a-5), (I-3b-1) ~ (I-3b-3), (I-4b-1) ~ (I- 4b-3), (1-5b-3), and (I-6b-1) had a docking score of -7 or less. On the other hand, the docking score of the control compound (X) was -6 or higher.

[Example 3]
(Docking simulation test for 3CL protease of SARS-CoV-1)
Then, instead of the co-crystal file of SARS-CoV-2 3CL protease and ligand, the same method as in Example 2 using the co-crystal file of SARS-CoV-1 3CL protease and ligand Among the compounds shown above, each compound (I-3a-1) to (I-3a-5), (I-4a-1) to (I-4a-5), (I-5a-4) , (I-6a-2) to (I-6a-4), (I-3b-1) to (I-3b-3), and (I-4b-1) to (I-4b-3) A docking simulation was performed.

The docking pose was similarly searched for the control compound (X) used in Example 1, and the docking score was calculated.

Compounds (I-3a-1) ~ (I-3a-5), (I-4a-1) ~ (I-4a-5), (I-5a-4), (I-6a-2) ~ ( In I-6a-4), (I-3b-1) to (I-3b-3), and (I-4b-1) to (I-4b-3), the docking score was -7 or less. . On the other hand, the docking score of the control compound (X) was -6 or higher.

[Example 4]
(Docking simulation test for 3CL protease of human rhinovirus)
Then, instead of the SARS-CoV-2 3CL protease and ligand co-crystal file, the human rhinovirus 3CL protease and ligand co-crystal file was used to replace CYS145SG with CYS147SG and HIS163NE. Using HIS161NE as a target, by the same method as in Example 2, among the compounds shown above, each compound (I-3a-1), (I-3a-2), (I-4a-1), (I -4a-5), (I-5a-1) ~ (I-5a-3), (I-6a-2) ~ (I-6a-5), (I-3b-1), (I-4b -2), (I-4b-3), (I-5b-2), (I-5b-23), and (I-6b-1) to (I-6b-2) docking simulations were performed .

Compounds (I-3a-1), (I-3a-2), (I-4a-1), (I-4a-5), (I-5a-1) ~ (I-5a-3), ( I-6a-2) ~ (I-6a-5), (I-3b-1), (I-4b-2), (I-4b-3), (I-5b-2), (I- 5b-3) and (I-6b-1) to (I-6b-2) had a docking score of -7 or less. On the other hand, the docking score of the control compound (X) was -6 or higher.

According to the compound of the present embodiment, it is possible to provide a novel compound that has inhibitory activity against protease and has a non-peptide-like structure. The protease inhibitor of this embodiment contains the above compound and can effectively inhibit proteases. The antiviral pharmaceutical composition of this embodiment contains the protease inhibitor and can effectively suppress viral replication and proliferation.

Claims (8)

A protease inhibitor containing a compound represented by the following general formula (I) as an active ingredient.

In general formula (I), R ¹¹ and R ¹² are each independently a hydrogen atom or a group represented by general formula (II) below, and when either one of R ¹¹ and R ¹² is a hydrogen atom, The other is a group represented by the following general formula (II). R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group, R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, —C(═O)—R″, or a cyano group, R′ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms or a benzyl group, and R″ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms. .

In general formula (II), an asterisk represents a binding site to a benzene ring, R ²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted or a group represented by the following general formula (III).

In general formula (III), an asterisk represents a binding site with an amide bond, Y ³¹ is a methylene group or a methylmethylene group, Q ³¹ is -NH- or -NH-C(=O)- and R ³¹ is a substituted or unsubstituted cyclohexyl group or a substituted or unsubstituted phenyl group. 2. The protease inhibitor according to claim 1, which contains a compound represented by the following general formula (I-1) or a compound represented by the following general formula (I-2) as an active ingredient.

In general formula (I-1), R ¹¹¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyrrolidinyl group, or the general formula (III) is a group represented by

In general formula (I-2), R ¹²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted pyrrolidinyl group, or the general formula (III) is a group represented by 3. The protease inhibitor according to claim 2, which contains a compound represented by the following formula (I-1-1) as an active ingredient.

The protease inhibitor according to any one of claims 1 to 3, wherein the protease is a cysteine protease. The protease inhibitor according to claim 4, wherein the protease is 3C protease or 3C-like protease. The protease inhibitor according to claim 5, wherein the protease is SARS-coronavirus-2 3C-like protease. An antiviral pharmaceutical composition containing the protease inhibitor according to any one of claims 1 to 3 and a pharmaceutically acceptable carrier. A compound represented by the following general formula (I).

In general formula (I), R ¹¹ and R ¹² are each independently a hydrogen atom or a group represented by general formula (II) below, and when either one of R ¹¹ and R ¹² is a hydrogen atom, The other is a group represented by the following general formula (II). R ¹³ is a 1H-imidazol-4-yl group or a pyrrolidin-2-one-3-yl group, R ¹⁴ is a formyl group, —CH═CHCO ₂ R′, —C(═O)—R″, or a cyano group, R′ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms or a benzyl group, and R″ is a substituted or unsubstituted alkyl group having 1 to 4 carbon atoms. .

In general formula (II), an asterisk represents a binding site to a benzene ring, R ²¹ is a substituted or unsubstituted alkyl group having 1 to 3 carbon atoms, a substituted or unsubstituted phenyl group, a substituted or unsubstituted or a group represented by the following general formula (III).

In general formula (III), an asterisk represents a binding site with an amide bond, Y ³¹ is a methylene group or a methylmethylene group, Q ³¹ is -NH- or -NH-C(=O)- and R ³¹ is a substituted or unsubstituted cyclohexyl group or a substituted or unsubstituted phenyl group.