JP2014080420A - Amino acid derivative with antifungal properties - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new antifungal compound and an amino acid derivative with antifungal properties.SOLUTION: This invention relates to a compound represented by NHR-CHR-C(O)-R(I) or salt thereof, and an antifungal agent including the compound or salt thereof.

Description

  The present invention relates to an antifungal amino acid derivative, and an antifungal agent, a food and drink, a cosmetic and a pharmaceutical composition containing the antifungal amino acid derivative.

Substances having antifungal properties are used as antifungal agents and antifungal treatments approved by the Food Sanitation Law. Known compounds having antifungal properties include imidazole compounds, diphenyl compounds, triazole compounds, candin compounds, etc. Among them, imidazole compounds and diphenyl compounds are used as food additives as antifungal agents. It is used. In addition, imidazole compounds, triazole compounds, and candin compounds are used as therapeutic agents for fungal infections.
Some antifungal amino acids have been found so far (Non-Patent Documents 1 to 5).

Molecular mode of action of the antifungal beta-amino acid BAY 10-8888, K. Ziegelbauer et al., Antimicrob. Agents Chemother., 42 (1998) 2197-2205. Novel antifungal beta-amino acids: synthesis and activity against Candida albicans, J. Mittendorf et al., Bioorg. Med. Chem. Lett. 13 (2003) 433-436. Alicyclic beta-amino acids in Medicinal Chemistry, A. Kuhl et al., Amino Acids, 29 (2005) 89-100. Antimicrobial activities of amino acid derivatives of monascus pigments, C. Kim et al., FEMS Microbiol Lett., 264 (2006) 117-124. Synthesis and antifungal activity of some 2-benzothiazolythioacetyl amino acid and peptide derivatives, A. Aboelmagd et al., In Proceedings of the 14th Int. Electron. Conf. Synth. Org. Chem., 1-30 November 2010; Sciforum Electornic Conferences Series , 2010, b019: 1-9.

  However, these conventional antifungal substances may lead to consumer anxiety due to restrictions on the amount and target of use, and there are those that are relatively difficult to synthesize and extract from natural products, In addition, there was a point to be improved, such as the need for synthesis with a controlled three-dimensional structure.

  Under such circumstances, there has been a demand for the development of an antifungal agent having characteristics such as no anxiety about safety or relatively easy synthesis.

  As a result of intensive studies to solve the above problems, the present inventors have found that an amino acid derivative containing a novel substance has antifungal activity. Since amino acids are naturally derived components, their derivatives are considered to have limited restrictions on the amount and target of use, and the synthesis of amino acid derivatives is relatively easy. It was thought that it could be an antifungal substance having characteristics that are not present. Based on these findings, the present inventor clarified that an antifungal amino acid derivative can be used as an antifungal agent and completed the present invention.

That is, the present invention relates to the following.
[1] General formula (I)
NHR ¹ —CHR ² —C (O) —R ³ (I)
[Where
R ¹ represents a hydrogen atom;
R ² represents an aryl C _1-6 alkyl group or a 5- to 10-membered heteroaryl C _1-6 alkyl group (wherein the aryl of the aryl C _1-6 alkyl group is a substituent represented by the formula (II)) The heteroaryl of the 5-10 membered heteroaryl C _1-6 alkyl group may have a substituent represented by the formula (II)), or R ¹ and R ² are combined A 5-membered non-aromatic heterocyclic group having a substituent of formula (II) may be formed;
-O-R ^2a (II)
(In the formula, R ^2a has an aryl C _1-6 alkyl group which may have a substituent, an aryl C _1-6 alkyl-OCO- which may have a substituent, and a substituent. An optionally substituted 5-10 membered heteroaryl C _1-6 alkyl group or an optionally substituted 5-10 membered heteroaryl C _1-6 alkyl-OCO-);
R ³ represents —NR ⁴ R ⁵ or —OR ⁶ ;
R ⁴ and R ⁵ are the same or different and each represents a hydrogen atom or a C _1-6 alkyl group (provided that one of R ⁴ and R ⁵ represents a group other than hydrogen);
R ⁶ represents a C _1-11 alkyl group. ]
Or a pharmaceutically acceptable salt thereof, an antifungal agent.
[2] General formula (Ib)
NHR ¹ —CHR ² —C (O) —R ^3b (Ib)
[Where
R ¹ represents a hydrogen atom;
R ² represents an aryl C _1-6 alkyl group or a 5- to 10-membered heteroaryl C _1-6 alkyl group (wherein the aryl of the aryl C _1-6 alkyl group is a substituent represented by the formula (II)) The heteroaryl of the 5-10 membered heteroaryl C _1-6 alkyl group may have a substituent represented by the formula (II)), or R ¹ and R ² are combined A 5-membered non-aromatic heterocyclic group having a substituent of formula (II) may be formed;
-O-R ^2a (II)
(In the formula, R ^2a has an aryl C _1-6 alkyl group which may have a substituent, an aryl C _1-6 alkyl-OCO- which may have a substituent, and a substituent. An optionally substituted 5-10 membered heteroaryl C _1-6 alkyl group or an optionally substituted 5-10 membered heteroaryl C _1-6 alkyl-OCO-);
R ^3b represents -NR ^4b R ^5b ;
R ^4b and R ^5b are the same or different and each represents a hydrogen atom or — (CH ₂ ) _m COOR ⁷ (provided that one of R ^4b and R ^5b represents a group other than hydrogen).
(In the formula, m represents an integer of 0 to 12, and R ⁷ represents a hydrogen atom or a C _1-6 alkyl group optionally having a substituent). ]
Or a pharmaceutically acceptable salt thereof, an antifungal agent.
[3] R ¹ is a hydrogen atom, R ² is indolyl-CH _2- , or a benzyl group having a substituent represented by the formula (II),
In Formula (II), R ^2a may have a substituent, a benzyl group, an optionally substituted benzyl-OCO—, or an optionally substituted pyridyl-CH ₂ —. or have a substituent is a pyridyl -CH ₂ -OCO-, antifungal agent according to [1] or [2].
[4] R ¹ and R ² together form a pyrrolidinyl group having a substituent represented by the formula (II),
In Formula (II), R ^2a may have a substituent, a benzyl group, an optionally substituted benzyl-OCO—, or an optionally substituted pyridyl-CH ₂ —. or have a substituent is a pyridyl -CH ₂ -OCO-, antifungal agent according to [1] or [2].
[5] The following compound groups:

An antifungal agent comprising at least one compound selected from: or a pharmaceutically acceptable salt thereof.
[6] The following compound groups:

At least one compound selected from: or a pharmaceutically acceptable salt thereof.
[7] General formula (IIIb)

[Wherein, -AA ^b -OH represents -NH- (CH ₂ ) _p -COOH, and p represents an integer of 2 to 5. ]
Or a pharmaceutically acceptable salt thereof.
[8] The compound according to [6] or [7] or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is a hydrochloride.
[9] A food or drink comprising the compound defined in any one of [1] to [5], the compound according to claim 6 or 7, or a pharmaceutically acceptable salt thereof.
[10] A cosmetic comprising the compound according to any one of [1] to [5], the compound according to claim 6 or 7, or a pharmaceutically acceptable salt thereof.
[11] A pharmaceutical composition comprising the compound defined in any one of [1] to [5], the compound according to claim 6 or 7, or a pharmaceutically acceptable salt thereof.

  The present invention provides new antifungal compounds. The antifungal compound of the present invention can be synthesized relatively easily. Moreover, since the antifungal compound of this invention is an amino acid derivative, compared with the existing antifungal substance, there is little possibility that the usage-amount and use object will be restrict | limited. Therefore, the antifungal compound of the present invention is useful as an antifungal agent, food and drink, cosmetics, and antifungal pharmaceutical composition.

The spectrum of ¹ H-NMR of Boc-Tyr (Z) -OH is shown. ¹ shows the ¹ H-NMR spectrum of H-Tyr (Pym) -OEt. ¹ shows the ¹ H-NMR spectrum of Boc-Tyr-OH. It is a photograph which shows the mode of the petri dish at the time of antifungal activity measurement. The HPLC chart (A) and ¹ H-NMR spectrum (B) of H-Tyr (Bzl) -OPr (C ₃ ) are shown. It is a photograph which shows the mode of the petri dish at the time of antifungal activity measurement. It is a measurement result of the antifungal activity about carbon number. It is a figure which shows the structure of an amino acid derivative. The HPLC chart and ¹ H-NMR spectrum of H-Tyr (Bzl) -Gly-OH are shown. The HPLC chart and ¹ H-NMR spectrum of H-Tyr (Bzl) -β-Ala-OH are shown. The HPLC chart and ¹ H-NMR spectrum of H-Tyr (Bzl) -GABA-OH are shown. The HPLC chart and ¹ H-NMR spectrum of H-Tyr (Bzl) -Ava-OH are shown. The HPLC chart and ¹ H-NMR spectrum of H-Tyr (Bzl) -Aca-OH are shown. ¹ H-NMR spectrum of ^{H-Tyr (Bzl) -OBu i} . ¹ H-NMR spectrum of ^{H-Tyr (Bzl) -OBu s} . ¹ H-NMR spectrum of ^{H-Tyr (Bzl) -OBu t} . It is a photograph which shows the mode of the petri dish at the time of antifungal activity measurement.

Hereinafter, the present invention will be described in detail.
It should be noted that all publications cited in the present specification, for example, prior art documents, publications, patent publications and other patent documents are incorporated herein by reference.

1. Summary of the Invention The present inventors have found that amino acid derivatives, particularly compounds synthesized from tyrosine, tryptophan or hydroxyproline, have antifungal activity and have completed the present invention.

  Since the amino acid derivative of the present invention is a compound derived from an amino acid, there is less concern about side effects compared to conventional antifungal agents, and it can be used in a wide range and may be used in a large amount. Furthermore, the amino acid derivative of the present invention can be chemically synthesized relatively easily.

  Therefore, the amino acid derivative of the present invention can be used as an active ingredient of an antifungal agent or an antifungal pharmaceutical composition. Moreover, the amino acid derivative of the present invention can be used in foods and drinks and cosmetics. When used in foods and drinks and cosmetics, it can also be used as an additive having an antifungal action.

2. Amino acid derivative In the present specification, an amino acid derivative is not particularly limited as long as it is a compound derived from an amino acid and has an antifungal action. The amino acid may be any of amino acids such as α-, β-, and γ-, but α-amino acids are preferred. The amino acid derivative of the present invention is, for example, a compound having tyrosine, tryptophan or hydroxyproline and having an antifungal action (see below).

  In addition, the amino acid derivative of the present invention may be a molecule in which two or more amino acids or amino acid derivatives are linked by peptide bonds, such as a dipeptide, tripeptide, tetrapeptide and the like.

  As the amino acid derivative in the present invention, a compound represented by the following general formula (I) (hereinafter also referred to as “the compound of the present invention”) or a pharmaceutically acceptable salt thereof can be used.

Formula (I)
NHR ¹ —CHR ² —C (O) —R ³ (I)

In general formula (I),
R ¹ represents a hydrogen atom;
R ² represents an aryl C _1-6 alkyl group or a 5- to 10-membered heteroaryl C _1-6 alkyl group (wherein the aryl of the aryl C _1-6 alkyl group is a substituent represented by the formula (II)) Or a heteroaryl of a 5 to 10-membered heteroaryl C _1-6 alkyl group may have a substituent represented by the formula (II)), or R ¹ and R ² are integrated To form a 5-membered non-aromatic heterocyclic group having a substituent of formula (II);
-O-R ^2a (II)
(In the formula, R ^2a has an aryl C _1-6 alkyl group which may have a substituent, an aryl C _1-6 alkyl-OCO- which may have a substituent, and a substituent. An optionally substituted 5-10 membered heteroaryl C _1-6 alkyl group or an optionally substituted 5-10 membered heteroaryl C _1-6 alkyl-OCO-);
R ³ represents —NR ⁴ R ⁵ or —OR ⁶ ;
R ⁴ and R ⁵ are the same or different and each represents a hydrogen atom or a C _1-6 alkyl group (provided that one of R ⁴ and R ⁵ represents a group other than hydrogen);
R ⁶ represents a C _1-11 alkyl group.

  As an amino acid derivative in another embodiment of the present invention, a compound represented by the following general formula (Ib) or a pharmaceutically acceptable salt thereof can be used. In the present specification, the compound represented by the general formula (Ib) is also a “compound of the present invention”.

Formula (Ib)
NHR ¹ —CHR ² —C (O) —R ^3b (Ib)

In general formula (Ib),
R ¹ and R ² are as previously defined herein;
R ^3b represents -NR ^4b R ^5b ;
R ^4b and R ^5b are the same or different and each represents a hydrogen atom or — (CH ₂ ) _m COOR ⁷ (provided that one of R ^4b and R ^5b represents a group other than hydrogen); 12 represents an integer, and R ⁷ represents a hydrogen atom or a C _1-6 alkyl group which may have a substituent.

In the present invention, a C _1-6 alkyl group, an aryl C _1-6 alkyl group, an aryl C _1-6 alkyl-OCO—, a 5-10 membered heteroaryl C _1-6 alkyl group or a 5-10 membered heteroaryl C _{1 Although the} substituent which -6 alkyl-OCO- may have is not necessarily limited, For example, 1 or more selected from a halogen atom (a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, etc.) is mentioned. be able to. Moreover, it is preferable to have a substituent in an aryl part or 5-10 membered heteroaryl.

  In the present specification, the “aryl group” means an aromatic hydrocarbon cyclic group, preferably an aryl group having 6 to 10 carbon atoms, such as a phenyl group, 1-naphthyl group, 2 -A naphthyl group, an indenyl group, an azulenyl group, etc. are mention | raise | lifted. A phenyl group is preferred.

  In the present specification, the “heteroatom” means a nitrogen atom, an oxygen atom or a sulfur atom. Preferably, it is a nitrogen atom.

  In the present specification, the “5- to 10-membered heteroaryl group” means a fragrance having 5 to 10 atoms constituting a ring and containing 1 to 5 heteroatoms in the atoms constituting the ring. Meaning cyclic group, and specific examples include furyl, thienyl, pyrrolyl, imidazolyl, triazolyl, tetrazolyl, thiazolyl, pyrazolyl, oxazolyl, isoxazolyl, isothiazolyl, furazanyl , Thiadiazolyl, oxadiazolyl, pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl, triazinyl, purinyl, pteridinyl, quinolyl, isoquinolyl, naphthyridinyl, quinoxalinyl, cinnolinyl, quinazolinyl, imidazolinyl, Pyridyl group, imidazothiazolyl group, imidazo Xazolyl group, benzothiazolyl group, benzoxazolyl group, benzimidazolyl group, indolyl group, isoindolyl group, indazolyl group, pyrrolopyridyl group, thienopyridyl group, furopyridyl group, benzothiadiazolyl group, benzooxadiazolyl group, pyridopyrimidinyl group Benzofuryl group, benzothienyl group, thienofuryl group and the like. Preferred 5-10 membered heteroaryl groups are pyridyl groups or indolyl groups.

In the present specification, the “C _1-6 alkyl group” represents a linear or branched alkyl group having 1 to 6 carbon atoms, and suitable groups include, for example, a methyl group, an ethyl group, an n-propyl group, iso-propyl group, n-butyl group, iso-butyl group, sec-butyl group, tert-butyl group, n-pentyl group, 1,1-dimethylpropyl group, 1,2-dimethylpropyl group, 2,2- Dimethylpropyl group, 1-ethylpropyl group, 2-ethylpropyl group, 1-methylbutyl group, 2-methylbutyl group, n-hexyl group, 1-methyl-2-ethylpropyl group, 1-ethyl-2-methylpropyl group 1,1,2-trimethylpropyl group, 1-propylpropyl group, 1,1-dimethylbutyl group, 1,2-dimethylbutyl group, 2,2-dimethylbutyl group, 1,3-dimethylbuty Group, 2,3-dimethylbutyl group, 2-ethylbutyl group, 2-methylpentyl group, 3-methylpentyl group and the like, more preferably methyl group, ethyl group, n-propyl group, iso-propyl group N-butyl group, iso-butyl group, sec-butyl group, tert-butyl group, n-pentyl group and the like.

In the present specification, the “aryl C _1-6 alkyl group” means a C _1-6 alkyl group substituted with an aryl group.

In the present specification, the “5- to 10-membered heteroaryl C _1-6 alkyl group” means a C _1-6 alkyl group substituted with a 5- to 10-membered heteroaryl group.

  In the present specification, the “5-membered non-aromatic heterocyclic group” contains 1 to 2 heteroatoms in the atoms constituting the ring, and may contain one double bond in the ring. Means a monocyclic non-aromatic cyclic group. Examples of the 5-membered non-aromatic heterocyclic group include a pyrrolidinyl group, an oxazolinyl group, an imidazolidinyl group, an oxazolidinyl group, a tetrahydrofuryl group, a dioxolanyl group, a tetrahydrothienyl group, a thiazolidinyl group, and a pyrrolidinyl group is preferable.

  In the present specification, “n-” means normal type or primary substituent, “sec-” means secondary substituent, and “t- (tert-)” Means a tertiary substituent, and “i- (iso-)” means an isotype substituent.

In the present invention, the compound of the present invention in which R ² is an aryl C _1-6 alkyl group preferably has a structure derived from tyrosine. Moreover, it is preferable that the compound of this invention whose R < ² > is a 5-10 membered heteroaryl _C1-6 alkyl group has a structure induced | guided | derived from tryptophan. In addition, the compound of the present invention in which R ¹ and R ² are combined to form a 5-membered non-aromatic heterocyclic group having a substituent represented by the formula (II) has a structure derived from hydroxyproline. It is preferable to have.

In one embodiment of the present invention, preferably, R ¹ is a hydrogen atom, and R ² is indolyl-CH ₂ — or a benzyl group having a substituent represented by the formula (II).

In another embodiment of the present invention, R ¹ and R ² are preferably combined to form a pyrrolidinyl group having a substituent represented by the formula (II). In this case, the nitrogen atom to which R ¹ is bonded corresponds to the nitrogen atom of the pyrrolidinyl group.

In R ² , the aryl in the aryl C _1-6 alkyl group or the 5-10 membered heteroaryl in the 5-10 membered heteroaryl C _1-6 alkyl group has a substituent represented by the formula (II) with respect to the alkyl group. And preferably in the para position. In R ¹ and R ² , the 5-membered non-aromatic heterocyclic group may have a substituent represented by the formula (II) at the 3-position counting from the terminal nitrogen atom of the formula (I). preferable.

In the formula (II), preferably, aryl is a phenyl group (Ph), and 5- to 10-membered heteroaryl is a pyridyl group. ^{That, R 2a} is preferably, Ph-CH ₂ - (benzyl _group), Ph-CH 2 _-OCO-, pyridyl -CH ₂ - or pyridyl _-CH 2 -OCO-. In formula (II), the aryl or pyridyl group may be substituted with one or more substituents. In the present invention, the preferred substituent of aryl or 5- to 10-membered heteroaryl in formula (II) is selected from a halogen atom, preferably a chlorine atom.

In the present invention, R ³ is preferably —NHCH ₃ or —OC _1-6 alkyl group. More preferably, R ³ is —OC ₄ H ₉ .

In the present invention, R ^3b is preferably —NH (CH ₂ ) _n —COOH (n represents an integer of 1 to 5).

In one embodiment of the present invention, the compound of the present invention is preferably a compound wherein R ¹ is a hydrogen atom and R ² is indolyl-CH ₂ —. R ³ is preferably —NHCH ₃ or —OC _1-6 alkyl group. The indolyl group preferably has a bond at the 3-position.

In another embodiment of the present invention, the compound of the present invention is preferably a compound wherein R ¹ is a hydrogen atom and R ² is a benzyl group having a substituent represented by the formula (II). In formula (II), aryl is a phenyl group, and 5- to 10-membered heteroaryl is a pyridyl group. The pyridyl group preferably has a bond at the 4-position. The phenyl or pyridyl group of formula (II) may be substituted with one or more halogen atoms. R ³ is preferably —NHCH ₃ or —OC _1-6 alkyl group.

In another embodiment of the present invention, a preferred compound of the general formula (I) is a compound in which R ¹ and R ² are combined to form a pyrrolidinyl group having a substituent represented by the formula (II). is there. In formula (II), aryl is a phenyl group, and 5- to 10-membered heteroaryl is a pyridyl group. The phenyl or pyridyl group of formula (II) may be substituted with one or more halogen atoms. R ³ is preferably —NHCH ₃ or —OC _1-6 alkyl group.

In another embodiment of the present invention, a preferred compound of the general formula (Ib) is preferably a compound wherein R ¹ is a hydrogen atom and R ² is a benzyl group having a substituent represented by the formula (II). In formula (II), aryl is a phenyl group, and 5- to 10-membered heteroaryl is a pyridyl group. The pyridyl group preferably has a bond at the 4-position. The phenyl or pyridyl group of formula (II) may be substituted with one or more halogen atoms. R ^3b is preferably a group represented by —NH (CH ₂ ) _n —COOH (n represents an integer of 1 to 5).

  The compound of the present invention or a pharmaceutically acceptable salt thereof is preferably a compound represented by the following or a pharmaceutically acceptable salt thereof.

The aromatic ring in R ^a may have one or more substituents (for example, a halogen atom). “Bzl” represents a benzyl group, “Z” represents a benzyloxy-carbonyl group, “Pym” represents a pyridin-4-yl-methyl group, and “Pym-OCO” represents pyridine- It represents a 4-yl-methoxycarbonyl group.

The aromatic ring in R ^a may have one or more substituents (for example, a halogen atom). “Bzl” represents a benzyl group, “Z” represents a benzyloxy-carbonyl group, “Pym” represents a pyridin-4-yl-methyl group, and “Pym-OCO” represents pyridine- It represents a 4-yl-methoxycarbonyl group.

In this specification, —C ₃ H ₇ is an n-propyl group or an iso-propyl group, and —C ₄ H ₉ is an n-butyl group, an iso-butyl group, a sec-butyl group, or a tert-butyl group. It is. The same applies to other groups represented by —C _n H _{2n + 1} , and may mean one or more of a plurality of isomers that may exist.

The compound of the present invention or a pharmaceutically acceptable salt thereof is more preferably a compound selected from the following compound group or a salt thereof.

Further, the compound of the present invention or a pharmaceutically acceptable salt thereof is more preferably a compound represented by the following formula (III) or a salt thereof (for example, hydrochloride).

Wherein, -AA-OH represents _{-NH- (CH 2) n -COOH,} n is an integer of 1 to 5. ]

  The compound represented by the formula (III) can be prepared by a conventional method using an amino acid and a tyrosine derivative Boc-Tyr (Bzl) -OH as raw materials. For example, when glycine is used as the amino acid, a compound with m = 1 is prepared; when β-alanine is used, a compound with m = 2 is prepared; when GABA (γ-aminobutyric acid) is used. M is 3; when Ava (5-aminovaleric acid) is used, m is 4; when Aca (ε-aminocaproic acid) is used, m is 5 A compound is made. In the present specification, compounds represented by the formula (III) in which m is 1 to 5 are respectively represented by H-Tyr (Bzl) -Gly-OH, H-Tyr (Bzl) -β-Ala-OH, H-Tyr ( Bzl) -GABA-OH, H-Tyr (Bzl) -Ava-OH, and H-Tyr (Bzl) -Aca-OH.

In another embodiment of the present invention, the compound of the present invention or a pharmaceutically acceptable salt thereof is at least one compound selected from the following compound group or a salt thereof (for example, hydrochloride). The present invention also includes these compounds or pharmaceutically acceptable salts thereof.

In the above compounds, _-C 3 _{H 7} preferably is n- propyl group, _-C 4 _{H 9} is preferably n- butyl or sec- butyl _group, -C _{5 H 11} is n- pentyl group.

The present invention also provides a compound of the general formula (IIIb)

[Wherein, -AA ^b -OH represents -NH- (CH ₂ ) _p -COOH, and p represents an integer of 2 to 5. ]
Or a pharmaceutically acceptable salt thereof.

  In the present invention, the general formula (I) or (Ib) includes the above-mentioned compounds, and the amino acid derivatives of the present invention are combined with one or several of these compounds or a pharmaceutically acceptable salt thereof. Can be used as

  Those skilled in the art can produce the amino acid derivative used in the present invention by a known method. For example, “Basics and Experiments of Peptide Synthesis (Nobuo Izumiya, Tetsuo Kato, Tomohiko Aoyagi, Michinori Waki) Maruzen Co., Ltd.”, “Biochemistry Experiment Course Volume 1 (Japan Biochemical Society) Tokyo Chemistry Doujin”, “Experiment It can be produced by the method described in “Chemical Course 16 (Edited by Chemical Society of Japan) Maruzen Co., Ltd.”.

Moreover, the amino acid derivative of this invention can be manufactured by the method as described in an Example as described in the following literature, for example. For example,
Boc-Tyr (Z) -OH: D. Yamashiro et al., J. Org. Chem., 38,591 (1973);
Boc-Tyr (Pym) -OH: Y. Nishiyama et al., Chem. Pharm. Bull., 49, 233 (2001);
Synthesis method of ester: Cs method-SS Wang et al., J. Org. Chem., 42, 1286 (1977).
NaHCO ₃ method-V. Bocchi et al., Synthesis, 1979, 961 .; and de-Boc: K. Sato et al., Bull. Chem. Soc. Jpn., 50, 1999 (1977).

  The amino acid derivative of the present invention may form a salt with an acid or a base. The amino acid derivatives in the present invention also include these salts. In the present specification, the salt is a pharmaceutically acceptable salt and is not particularly limited as long as it forms a pharmaceutically acceptable salt with an amino acid derivative. For example, hydrochloride, hydrobromide, Inorganic acid addition salts such as sulfate or phosphate, organic acid salts such as acetate, propionate, citrate, tartrate, malate, or oxalate, or sodium, potassium, or calcium And salts such as salts. For those skilled in the art, salts of the amino acid derivatives of the present invention are suitable acids such as hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, oxalic acid, fumaric acid, methanesulfonic acid, or sodium hydroxide, potassium hydroxide, ammonia, etc. Can be prepared using any suitable base.

  In the present invention, the amino acid derivative has an asymmetric carbon depending on the type of the substituent, and optical isomers may exist. These optical isomers are also included in the amino acid derivative of the present invention.

3. Antifungal Agent The antifungal agent of the present invention contains the amino acid derivative of the present invention.

  In the present specification, the fungus is a eukaryote classified as a fungus (Eumycota or Eumytes), and is not particularly limited. In general, mushrooms, molds, yeasts and the like are known. In the present invention, the fungus is, for example, apple rot (Valsa ceratosperma Maire), white rot (Rosellinia necatrix Prill.), Gray mold (Botrytis cinerea), rice blight (Fusarium solani, Rhizopus oryzae, Trichoderma viride) ), Penicillium sp., NeurosporAcrassa, Monascus purpureus, and Aspergillus fumigatus, but are not limited thereto.

  Examples of the antifungal agent of the present invention include, but are not limited to, apple rot (Valsa ceratosperma Maire), white rot (Rosellinia necatrix Prill.), Gray mold (Botrytis cinerea), and rice wilt (Fusarium solani, Rhizopus oryzae, Trichoderma viride), blue mold (Penicillium sp.), Red spider mold (NeurosporAcrassa), white mold fungus (Monascus purpureus), and at least one fungus selected from the group consisting of Aspergillus fumigatus (Aspergillus fumigatus), etc. Antifungal activity.

  The antifungal action is an action that suppresses the growth of the fungus, and stops the growth of the fungus, reduces the growth rate of the fungus, delays the start of the growth of the fungus, prevents or treats the disease derived from the fungus (reduction of symptoms, stop, Including delay of onset, prevention of recurrence, etc.).

  Since the amino acid derivative of the present invention is a derivative from an amino acid, it is less likely to limit the amount used or the target of use compared to existing imidazole compounds and diphenyl antifungal substances. Therefore, the antifungal agent of the present invention has less fear of side effects compared to existing antifungal agents, and can be used in a wide range, and can also be used in various doses.

  The antifungal agent of the present invention can be used for inhibiting fungal growth. In one embodiment of the present invention, the antifungal of the present invention is applied to seeds, flowers, leaves, stems or roots of plants in order to prevent or treat diseases derived from fungi of plants (eg, wheat, barley, or rose). Apply the agent. In another aspect of the present invention, in order to prevent or treat diseases derived from fungi in animals (for example, mammals such as humans, rats, rabbits, sheep, pigs, cattle, cats, dogs, monkeys, etc.) The antifungal agent of the present invention is applied to animals orally. In one aspect of the invention, the animal is a mammal other than a human. In yet another aspect of the present invention, the present invention may be applied to the surface or interior of a substrate to inhibit fungal growth on the substrate (eg, paint, paper, wood, leather, concrete, plastic, or metal). Apply antifungal agent. In yet another aspect of the present invention, the antifungal agent of the present invention is applied to a cell culture system in order to suppress fungal growth in the cell culture system.

  “Application” includes administration, spraying, application, contact, infusion, standing, and the like.

  Further, the antifungal agent of the present invention may contain a carrier or an additive in addition to the amino acid derivative of the present invention, depending on the purpose of use. Such carriers and additives include water, acetic acid, organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin , Methylcellulose, ethylcellulose, xanthan gum, gum arabic, casein, agar, polyethylene glycol, diglycerin, glycerin, propylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, surfactant, etc. Can be mentioned.

  The amount of the amino acid derivative of the present invention used in the antifungal agent of the present invention varies depending on the purpose of use. A person skilled in the art can appropriately select the amount of the amino acid derivative of the present invention to be used according to the purpose of use.

  For example, when the antifungal agent of the present invention is used to prevent or treat a disease derived from a fungus of a plant, the amount of the amino acid derivative of the present invention used is, for example, 0 per day for a 100 g plant. 0.001 mg to 1000 mg.

  When the antifungal agent of the present invention is used for preventing or treating diseases derived from fungi of animals, the amount of the amino acid derivative of the present invention used is, for example, 0.01 kg per day for 60 kg animals. -1000 mg, or 0.05-500 mg.

  When the antifungal agent of the present invention is used to suppress fungal growth on the substrate, the amount of the amino acid derivative of the present invention used is, for example, 0.001 μg to 1000 mg in the case of a 50 g substrate. .

  When the antifungal agent of the present invention is used to suppress fungal growth in a cell culture system, the amino acid derivative of the present invention is used in an amount of 50 g of medium (for example, the weight excluding cultured cells). For example, it is 0.001 microgram-1000 mg.

  The present invention also provides amino acid derivatives for antifungal agents or amino acid derivatives for inhibiting fungal growth. The fungi whose growth can be inhibited by the amino acid derivative of the present invention, and the usage and dosage of the amino acid derivative of the present invention are the same as those described for the antifungal agent.

  The present invention also provides a method for inhibiting fungal growth using the amino acid derivative of the present invention. Here, "using the amino acid derivative of the present invention" means, for example, the amino acid derivative of the present invention on seeds, flowers, leaves, stems or roots of plants in order to prevent or treat diseases derived from fungi of plants. Application of the amino acid derivative of the present invention to animals orally or parenterally to prevent or treat fungal-derived diseases in animals (for example, excluding humans), growth of fungi on a substrate In order to suppress the growth, the amino acid derivative of the present invention is applied to the surface or inside of the substrate, or the amino acid derivative of the present invention is applied to the cell culture system in order to suppress fungal growth in the cell culture system. Including, but not limited to. The fungi capable of inhibiting the growth by the method of the present invention, and the usage and dosage of the amino acid derivative of the present invention are the same as described for the antifungal agent.

  Furthermore, the present invention provides the use of the amino acid derivative of the present invention for the production of an antifungal agent. The fungi capable of inhibiting the growth by use of the amino acid derivative of the present invention, and the usage and dosage of the amino acid derivative of the present invention are the same as those described for the antifungal agent.

4). Food / Beverage The present invention provides a food / beverage containing the amino acid derivative of the present invention. By adding the amino acid derivative of the present invention, fungal growth in food and drink can be suppressed. The food and drink of the present invention include, for example, beverages (coffee, cocoa, oolong tea, black tea, green tea, fruit beverages, vegetable juice, milk, etc.), alcoholic beverages (beer, sake, shochu, whiskey, vodka, gin, brandy, rum , Liqueur, wine, laochu or plum wine), seasonings (such as miso, sauce, sugar, salt, soy sauce, vinegar or spices), confectionery (candy, ice cream, caramel, cookies, cakes, pie, jelly, chocolate or Waffles, etc.), cooked rice, buckwheat, buckwheat noodles, udon, bread, pizza, cream, cheese, butter, jam, edible oils and fats (such as olive oil or corn oil) or other processed foods and beverages. Add the amino acid derivative.

  The addition amount of the amino acid derivative of the present invention with respect to the food and drink of the present invention is, for example, about 0.001 wt% to 99 wt% with respect to the total weight of the food and drink of the present invention.

  Moreover, other additives, such as a gelatinizer or an emulsifier, can also be added to the food / beverage of this invention as needed.

  The present invention provides a method for inhibiting fungal growth in food and drink using the amino acid derivative of the present invention. Here, “using the amino acid derivative of the present invention” means adding the amino acid derivative of the present invention to a food or drink, for example, mixing the amino acid derivative of the present invention with a food or drink, or the surface of a food or drink And the like (for example, coating, spraying) and the like. The food and drink to which the amino acid derivative of the present invention is added and the amount added are the same as those described in the above food and drink.

5. Cosmetics The present invention provides cosmetics containing the amino acid derivative of the present invention. By adding the amino acid derivative of the present invention, fungal growth in cosmetics can be suppressed. The cosmetics of the present invention include, for example, cosmetic soaps, shampoos, facial cleansers, rinses, eye creams, eye shadows, creams, emulsions, skin lotions, perfumes, funnier, cosmetic oils, cosmetics for hair, hair dyes, perfumes, powders. , Pack, shaving cream, shaving lotion, suntan oil, sunscreen oil, suntan lotion, sunscreen lotion, sunscreen cream, sunscreen cream, foundation, powdered perfume, blusher, mascara, eyebrow, nail cream, beauty nail enamel , Beauty nail enamel remover, hair wash, bath cosmetic, lipstick, lip balm, eyeliner, toothpaste, deodorant, eau de cologne, hair nourishing agent, hair restorer, etc., and the amino acid derivative of the present invention is added to these cosmetics .

  The amount of the amino acid derivative of the present invention added to the cosmetic of the present invention is, for example, about 0.001% to 99% by weight with respect to the total weight of the cosmetic of the present invention.

  Various ingredients other than the amino acid derivative of the present invention can be further added to the cosmetic of the present invention depending on the purpose of use. For example, improvement of emollient effect, improvement of feeling of use, reduction of bulk after use, improvement of solubility, improvement of emulsification, improvement of emulsification stability, improvement of compatibility with oil components, reduction of feeling of tension after use, improvement of skin familiarity, skin Improvement of spread, reduction of stickiness, prevention of rough skin, improvement of skin effect, improvement of skin protection effect, improvement of keratin, normalization of skin keratinization (preventing keratinization due to increased skin turnover, prevention of skin thickening, epidermal lipid metabolism Abnormal Suppression), reduction of dry skin such as senile xerosis, improvement of dry skin such as cracks and desquamation, suppression of wrinkle generation, disappearance of wrinkles, wound treatment, prevention and improvement of pigmentation, prevention of aging, reduction of dandruff and itching Suitable ingredients for the purpose of reducing hair loss, preventing and treating scalp disease, improving storage stability, improving flexibility, improving elasticity, imparting gloss, suppressing melanin pigment production, or preventing sunburn It can be pressurized.

  As components that can be added to the cosmetic of the present invention, for example, oil and fat components, phospholipids, UV absorbers, IR absorbers, emulsifiers, surfactants, preservatives, antifungal agents, antioxidants, whitening agents, vitamins, Amino acids, hormones, peptides, bioactive plant extracts, fluorescent materials, pigments, dyes, fragrances, scrub agents, sequestering agents, binders, extenders, thickeners, sugars, nutrients, pH regulators, chelating agents, Bactericides, keratin improvers, keratolytic agents, antibiotics, skin permeation enhancers, blood circulation enhancers, anti-inflammatory agents, cell activators, anti-inflammatory agents, analgesics, emollients, emollients, wound treatment agents, or Metabolism promoters and the like can be appropriately blended depending on the purpose of use.

  The present invention provides a method for inhibiting fungal growth in cosmetics using the amino acid derivative of the present invention. Here, “using the amino acid derivative of the present invention” means adding the amino acid derivative of the present invention to cosmetics, for example, mixing the amino acid derivative of the present invention into cosmetics, or Application of the amino acid derivative of the invention (application, spraying, etc.) is included. The cosmetic to which the amino acid derivative of the present invention is added and the amount added are the same as those described for the cosmetic.

6). Pharmaceutical Composition The present invention provides a pharmaceutical composition containing the amino acid derivative of the present invention. As described above, the salt in the amino acid derivative of the present invention is a pharmaceutically acceptable salt. Since the amino acid derivative of the present invention has antifungal activity, administration of the pharmaceutical composition of the present invention containing the amino acid derivative to a subject causes diseases such as mycosis caused by the body surface of the subject or a fungus in the body. Can be treated. Mycosis includes superficial mycosis or deep mycosis. More specifically, athlete's foot or candidiasis (such as oral candidiasis, esophageal candidiasis, or gastrointestinal candidiasis) can be mentioned.

  The subject to which the pharmaceutical composition of the present invention is administered is a mammal such as a human, rat, rabbit, sheep, pig, cow, cat, dog, monkey or the like, preferably a human or a non-human mammal.

  The pharmaceutical composition of the present invention can employ both oral and parenteral dosage forms.

  The dosage form may be the amino acid derivative of the present invention used as it is, or a pharmaceutical preparation containing a pharmaceutically acceptable carrier or additive in addition to the amino acid derivative of the present invention according to a conventional method. May be. Such carriers and additives include water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water-soluble dextran, carboxymethyl. Sodium starch, pectin, methylcellulose, ethylcellulose, xanthan gum, gum arabic, casein, agar, polyethylene glycol, diglycerin, glycerin, propylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, pharmaceutical Examples of acceptable surfactants include additives.

  The above additives are selected from the above alone or in appropriate combination depending on the dosage form of the pharmaceutical composition of the present invention. As for the dosage form, in the case of oral administration, it can be administered as a tablet, capsule, fine granule, powder, granule, liquid, syrup or the like, or in an appropriate dosage form. In the case of parenteral administration, injection dosage forms and the like can be mentioned. In the case of an injection dosage form, it can be administered systemically or locally by intravenous injection such as infusion.

  For example, when used as an injectable preparation, the pharmaceutical composition of the present invention is dissolved in a solvent (for example, physiological saline, buffer solution, glucose solution, etc.), and an appropriate additive (human serum albumin, etc.) is added thereto. Things can be used. Alternatively, it may be freeze-dried to obtain a dosage form that dissolves before use. As the freeze-drying excipient, for example, sugar alcohols and saccharides such as mannitol and glucose can be used.

  The dosage of the pharmaceutical composition of the present invention or the amino acid derivative of the present invention varies depending on age, sex, symptom, administration route, administration frequency, and dosage form. The administration method is appropriately selected depending on the age and symptoms of the patient. For example, in the case of an adult (60 kg), the dose is 0.01 to 1000 mg, or 0.05 to 500 mg per day. The administration method is appropriately selected depending on the age and symptoms of the patient. Administration may be divided, for example, once or 2-4 times per day.

  The present invention also provides a method for treating mycosis comprising administering the amino acid derivative of the present invention to a patient. Furthermore, the present invention provides the use of the amino acid derivative of the present invention for producing the pharmaceutical composition for treating mycosis of the present invention. The method for administering the amino acid derivative of the present invention in the method for treating mycosis is the same as that described in the above pharmaceutical composition. In addition, the method for producing a pharmaceutical composition in the use of the amino acid derivative of the present invention is the same as that described in the above pharmaceutical composition.

  Furthermore, the present invention provides a method for inhibiting fungal growth in a pharmaceutical composition using the amino acid derivative of the present invention. Here, “using the amino acid derivative of the present invention” means adding the amino acid derivative of the present invention to a pharmaceutical composition, for example, mixing the amino acid derivative of the present invention with a pharmaceutical composition, or a pharmaceutical composition Application of the amino acid derivative of the present invention to the surface of an object is included. The addition amount of the amino acid derivative of the present invention is, for example, about 0.01 wt% to 99 wt% with respect to the total weight of the pharmaceutical composition of the present invention.

Abbreviations used in this specification are as follows.
AcOEt: Ethyl acetate
Boc-: t-Butyloxycabonyl
Bzl-: Benzyl
Bzl-Cl ₂ : 2,6-Dichlorobenzyl
DCC: N, N'-Dicyclohexylcarbodiimide
DCUrea: N, N'-Dicyclohexylurea
DMAP: N, N-dimethyl-4-aminopyridine
DMF: N, N-dimethylformamide
DMSO: N, N-dimethyl sulfoxide
DOX: 1,4-Dioxane
HPLC: High performance liquid chromatography
IBCF: Isobutyl chloroformate
MA: Mixed anhydride
MeOH: Methenol
-NH ₂ : Amide
-NHCH ₃ : Methyl amide
NMM: N-Methylmorpholine
¹ H-NMR: Proton nuclear magnetic resonance
-OBzl: Benzyl ester
-OEt: Ethyl ester
-OMe: Methyl ester
-OPr: Propyl ester
-OBu: Butyl ester
-OPen: Pentyl ester
-OHex: Hexyl ester
-OLau: Lauryl ester
-OOct: Octyl ester
-OSu: Succinimide ester
PDA: Potato dextrose agar
TFA: trifluoroacetic acid
TLC: Thin-layer chromatography

  Hereinafter, the present invention will be specifically described by way of examples.

1. Synthesis of amino acid derivatives
H-Tyr (Z) -OEt, H-Tyr (Pym) -OEt, H-Try-OEt, and H-Hyp (Bzl) -OEt were synthesized by the following procedure (Schemes 1 and 2).

(1) Synthesis of Boc-Tyr (Z) -OH and Boc-Tyr (Pym) -OH (1-1) Boc-Tyr (Z) -OH
Boc-Tyr-OH 2 mmol (0.5142 g) (Watanabe Chemical Co., Ltd. (Hiroshima)) was dissolved in 5 mL of acetone, and 3.6 mL of 1 mol / L NaOH was added with stirring at 0 to 5 ° C. To this solution, 0.3945 g of an acetone solution of Z-OSu was added dropwise, then 0.0247 g of DMAP was added, and the mixture was further stirred overnight at room temperature. The pH of the reaction solution was adjusted to 2 with 10% citric acid and then concentrated under reduced pressure, and the product was extracted with AcOEt. The aqueous layer was further extracted twice with AcOEt. The AcOEt layers were combined, washed with water, and dehydrated with sodium sulfate. The next day, sodium sulfate was filtered off with AcOEt, concentrated under reduced pressure, and dried under reduced pressure overnight to obtain Boc-Tyr (Z) -OH.
(1-2) Boc-Tyr (Pym) -OH
Boc-Tyr-OH was dissolved in DMF, and sodium hydride was added to this solution at 0 ° C. with stirring. After the evolution of hydrogen gas ceased, 4- (Bromomethyl) pyridine Hydrobromide was added. The reaction was stirred at room temperature overnight. The progress of the reaction was confirmed by TLC. Water and AcOEt were put into the separatory funnel and shaken well to separate the aqueous layer and AcOEt. The aqueous layer was adjusted to pH 4 with citric acid, and AcOEt was added and shaken well to leave AcOEt. AcOEt was washed with saturated brine and dehydrated with sodium sulfate overnight. The next day, sodium sulfate was removed by filtration, AcOEt was concentrated under reduced pressure, and the oil was dried under reduced pressure to obtain Boc-Tyr (Pym) -OH.

(2) Synthesis of ethyl ester (2-1) Boc-Tyr (Z) -OEt
Boc-Tyr (Z) -OH synthesized in the above (1-1) was reacted with iodoethane to carry out ethylation of Boc-Tyr (Z) -OH. Boc-Tyr (Z) -OH 1.7 mmol (0.755 g) was dissolved in MeOH (3.4 mL), and H ₂ O (1.122 mL) and CsCO ₃ 0.85 mmol (0.2791 g) were added to synthesize an amino acid cesium salt. After concentrating the solution under reduced pressure, DMF (3.4 mL) and iodoethane (0.136 mL) were added to the residue, and the mixture was stirred overnight at low temperature and under light shielding. The completion of the reaction was confirmed by TLC, and precipitated CsBr was removed by filtration. After the product was extracted with AcOEt, the AcOEt layer was washed with water, NaHCO ₃ aqueous solution and water, and dehydrated with sodium sulfate overnight. After dehydration, sodium sulfate was removed by filtration, AcOEt was concentrated under reduced pressure, and dried under reduced pressure overnight to obtain Boc-Tyr (Z) -OEt.
(2-2) Boc-Tyr (Pym) -OEt
Boc-Tyr (Pym) -OH synthesized in (1-2) and iodoethane were reacted in the same manner as described in (2-1) to obtain Boc-Tyr (Pym) -OEt. .
(2-3) Boc-Trp-OEt
Boc-Trp-OH 1 mmol (0.3044 g) and NaHCO ₃ 2 mmol (0.1680 g) are suspended in 5 ml of DMF, a solution of 5 ml of iodoethane (0.3999 ml) in 5 ml of DMF is added, and the mixture is stirred at room temperature for 24 hours. It was. Water was added and extracted with AcOEt. The AcOEt layer was washed with water and dehydrated with sodium sulfate overnight. The next day, sodium sulfate was collected by filtration, concentrated under reduced pressure, and dried under reduced pressure to obtain Boc-Trp-OEt.
(2-4) Boc-Hyp (Bzl) -OEt
Boc-Hyp (Bzl) -OH and iodoethane were reacted in the same manner as described in (2-3) above to obtain Boc-Hyp (Bzl) -OEt.

(3) Removal of Boc group by HCl / dioxane (3-1) H-Tyr (Z) -OEt
Boc-Tyr (Z) -OEt obtained in (2-1) was dissolved in HCl / dioxane (2.125 mL) and placed at room temperature for 1 hour. N ₂ gas was introduced and solidified with diethyl ether to obtain H-Tyr (Z) -OEt hydrochloride.
(3-2) H-Tyr (Pym) -OEt
The Boc-Tyr (Pym) -OEt obtained in (2-2) is deprotected by the same method as described in (3-1) above, and H-Tyr (Pym) -OEt hydrochloride is obtained. Obtained.
(3-3) H-Trp-OEt
The Boc-Trp-OEt obtained in (2-3) was deprotected by the same method as described in (3-1) to obtain H-Trp-OEt hydrochloride.
(3-4) H-Hyp (Bzl) -OEt
Boc-Hyp (Bzl) -OEt obtained in (2-4) is deprotected by the same method as described in (3-1) above, and H-Hyp (Bzl) -OEt hydrochloride is obtained. Obtained.

Table 4 shows the yield and yield of each amino acid derivative synthesized as described above.

(4) Thin layer chromatography (TLC)
The progress of the reactions (1) to (3) was confirmed by TLC spots.
As TLC samples, reaction raw materials only, reaction raw materials + post-reaction samples, and post-reaction samples only are spotted with capillaries, developed using a developing solvent (chloroform: methanol = 19: 1), dried, and then a ninhydrin reagent. Was sprayed and heated.

(5) High performance liquid chromatography (HPLC)
The column used for HPLC was 5C ₁₈ -PAQ size 4.6 × 150 mm (Nacalai Tesque), the detection wavelength was 215 nm, the detection sensitivity was 1024 mV FS, and the chart speed was 5 mm / mim. Elution was performed using a water-acetonitrile-0.1% TFA system, and the acetonitrile content was linearly changed from 10 to 90% in 0 to 30 minutes. The flow rate was 1 ml / min. The temperature was 35 ° C. As a measurement sample, 1 mg of an amino acid derivative dissolved in 1000 μl of MeOH was used.

  The product obtained in the above (1) and (3) was analyzed by HPLC to confirm that the target amino acid derivative was obtained.

(6) Nuclear magnetic resonance ( ¹ H-NMR)
In the reactions (1) to (3), ¹ H-NMR was measured. ^It was confirmed from the ¹ H-NMR spectrum whether the desired product was obtained.
A sample for measurement was prepared by dissolving 7 to 10 mg of an amino acid derivative in CDCl ₃ 750 to 1000 μl. TMS (Tetramethyl silane) was used as an internal standard.

  The spectra of Boc-Tyr (Z) -OH, H-Tyr (Pym) -OEt, and Boc-Tyr-OH as a raw material are shown in FIGS. 1, 2, and 3, respectively.

1 and 3, it was confirmed that a Z group (carbobenzoxy group) could be introduced into Boc-Tyr-OH, and Boc-Tyr (Z) -OH could be synthesized.
Moreover, in FIG. 2, since the peak specific to pyridine of (i) was confirmed, it was confirmed that H-Tyr (Pym) OEt could be synthesized. The large peak of (ii) is a peak derived from water dissolved in DMSO.

Therefore, (5) high performance liquid chromatography (HPLC) and (6) nuclear magnetic resonance ( ¹ H-NMR) confirmed that the target product was synthesized.

  Further, H-Tyr (Bzl) -OEt and H-Tyr (Bzl) -OMe were synthesized by the same method as described above. H-Tyr (Bzl) -OEt was synthesized from Boc-Tyr (Bzl) -OH (commercially available product) with the ethyl ester Boc-Tyr (Bzl) -OEt, followed by removal of the Boc group and H-Tyr (Bzl) -OEt · HCl was obtained. H-Tyr (Bzl) -OMe was synthesized from Boc-Tyr (Bzl) -OH (commercially available product) with the methyl ester Boc-Tyr (Bzl) -OMe, followed by removal of the Boc group and H-Tyr (Bzl) -OMe · HCl was obtained.

2. Antifungal activity measurement
The antifungal activity of amino acid derivatives and standard samples against Valsa Ceratosperma Maire was measured.
First, potato dextrose agar (PDA) medium was heated and dissolved, cooled to about 50 ° C., and then spores of Valsa Ceratosperma Maire were suspended. 20 ml of this medium was spread in a petri dish and allowed to solidify at room temperature. Samples dissolved in DMSO to a concentration of 25 mg / ml were used as measurement samples, and 50 μl each was spotted on a paper disk in the medium. After culturing at 25 ° C. for 3 days, antifungal activity was observed. In addition, a standard sample dissolved in 5 mg / ml was used as a measurement sample. Antifungal activity is represented by the diameter of the blocking circle against Valsa Ceratosperma Maire.

The following samples were used for the measurement.
<Standard sample>
1. Biphenyl Hinokitiol 3. 3. Miconazole nitrate Benzyl phenyl ether 5. L-thyroxine <amino acid derivative>
6). H-Tyr (Bzl) -OEt
7). H-Tyr (Z) -OEt
8). H-Tyr (Pym) -OEt
9. H-Trp-OEt
10. H-Hyp (Bzl) -OEt

  Hinokitiol and miconazole nitrate are known compounds as antifungal agents. Antifungal activity was also measured for benzylphenyl ether having a structure similar to that of the Tyr derivative, biphenyl having two ring structures, and L-thyroxine.

The measurement results are shown in Table 5 and Table 6. Moreover, the photograph which shows the mode of the petri dish at the time of a measurement is shown in FIG.

  In Tables 5 and 6, the diameter of the inhibition circle of the standard sample prepared at a concentration of 5 mg / mL was corrected to a value at a concentration of 25 mg / mL. In the table, the diameter of the blocking circle is the diameter of the blocking circle including the paper disk, “−” is ˜10 mm, “+” is 10 to 20 mm, “++” is 20 to 30 mm, and “++++” is 30 mm ~ is shown.

  As a result, it was confirmed that hinokitiol and miconazole nitrate have strong antifungal activity, while biphenyl and L-thyroxine have no antifungal activity. Benzyl phenyl ether was also confirmed to have antifungal activity.

  Furthermore, H-Tyr (Bzl) -OEt, H-Tyr (Z) -OEt, H-Trp-OEt and H-Hyp (Bzl) -OEt were shown to have antifungal activity. H-Tyr (Pym) -OEt showed weak antifungal activity. H-Tyr (Bzl) -OMe was also shown to have antifungal activity.

The following can be considered from the measurement results of antifungal activity.
First, a compound containing a carboxyl group (-C (= O) O-) between the Bzl group and tyrosine of the ethyl ester of tyrosine (H-Tyr (Bzl) -OEt) whose side chain is protected with a benzyl group (H -Tyr (Z) -OEt) exhibits antifungal activity, indicating that the effect does not change even if another group is inserted between the aromatic ring and the aromatic ring.

It was shown that the Tyr derivative (H-Tyr (Pym) -OEt) in which the protecting group of the Tyr derivative also has antifungal activity. At first glance, the antifungal activity of H-Tyr (Pym) -OEt is lower than that of other derivatives. This is because the sample produced in this example has a low purity according to the HPLC peak, and thus the antifungal activity is low. The effect seems to have weakened.
Moreover, it was shown from the peak position of HPLC that hydrophilicity raises by N entering into the ring structure of a protecting group. Based on this finding, it is possible to provide a more hydrophilic antifungal substance.

Furthermore, this example showed that amino acid derivatives other than tyrosine derivatives also showed antifungal activity.
Ethyl ester of Trp (H-Trp-OEt) and ethyl ester of Hyp with side chain protected with Bzl (H-Hyp (Bzl) -OEt) also showed antifungal activity. From this, the presence or absence of antifungal activity may involve the flexibility of the side chain portion of the amino acid. In this example, biphenyl, which did not exhibit antifungal activity, has a low flexibility and therefore does not overlap with the aromatic ring. However, active compounds (benzylphenyl ether, H-Tyr (Z) -OEt, H-Tyr (Pym ) -OEt, H-Trp-OEt, and H-Hyp (Bzl) -OEt) are flexible, and when they have a plurality of aromatic rings, they can have a structure in which the aromatic rings overlap.

H-Tyr (Bzl) -OEt hydrochloride whose side chain protecting group is Bzl, H-Tyr (Bzl) -OMe hydrochloride, and BzlCl ₂ (2,6-dichloro-benzyl-) side chain protecting group Antifungal activity of H-Tyr (BzlCl ₂ ) -OEt hydrochloride and H-Tyr (BzlCl ₂ ) -OMe hydrochloride was measured by the same method as in Example 1.
As a result, the diameters of the inhibition circles against Valsa Ceratosperma Maire were not significantly different from each other, indicating that the antifungal activities were almost the same (H-Tyr (Bzl) -OMe hydrochloride had (+) , Otherwise (++)). Therefore, in the present invention, it has been clarified that the side chain protecting group of Tyr may not be substituted or may be substituted with a substituent such as halogen.

In this example, the relationship between the alkyl chain length of the C-terminal ester of the tyrosine derivative and the antifungal activity was examined. The antifungal activity was also examined when the C-terminal ester was changed to an alkylamide group such as —NHCH ₃ .

The tyrosine derivatives studied in this example are shown below.
・ H-Tyr (Bzl) -OMe (C ₁ )
・ H-Tyr (Bzl) -OEt (C ₂ )
・ H-Tyr (Bzl) -OPr (C ₃ )
・ H-Tyr (Bzl) -OBu (C ₄ )
・ H-Tyr (Bzl) -OPen (C ₅ )
・ H-Tyr (Bzl) -OHex (C ₆ )
・ H-Tyr (Bzl) -OLau (C ₁₂ )
・ H-Tyr (Bzl) -OOct (C ₁₈ )
・ H-Tyr (Bzl) -NH ₂ (NH ₂ )
・ H-Tyr (Bzl) -NHCH ₃ (NHCH ₃ )

1. Synthesis of amino acid derivative by Boc liquid phase method (1) Synthesis of amino acid benzyl ester A tyrosine derivative was synthesized by the following procedure (Scheme 3).

Boc tyrosine (2 mmol) was dissolved in MeOH (4 ml), H ₂ O (1.32 ml) and Cs ₂ CO ₃ (1 mmol) were added to synthesize an amino acid cesium salt. The solution was concentrated under reduced pressure, DMF (4 ml) and lauryl bromide (0.48 ml) were added to the residue, and the mixture was stirred at a low temperature. After reacting overnight, the completion of the reaction was confirmed by TLC, and precipitated CsBr was removed by filtration. After extraction with ethyl acetate, it was dehydrated overnight with sodium sulfate. After dehydration, sodium sulfate was removed by filtration, and ethyl acetate was concentrated under reduced pressure to obtain Boc amino acid lauryl ester. Before using the liquid phase method, de-Boc was performed with dioxane hydrochloride to obtain an ester hydrochloride.

  Similarly, H-Tyr (Bzl) -OPr (using 1-bromopropane), H-Tyr (Bzl) -OBu (using 1-bromobutane), H-Tyr (Bzl) -OPen (using 1-bromopentane) Use), H-Tyr (Bzl) -OHex (using 1-bromohexane), and H-Tyr (Bzl) -OOct (using 1-bromooctadecane).

(2) Mixed acid anhydride method (MA method)
Synthesis of tyrosine derivatives using the MA method followed Scheme 4.

  Boc amino acid was dissolved in DMF, cooled to −15 ° C. or lower with stirring under ice cooling, and NMM and IBCF were added. After stirring for 10 minutes, a mixed solution of amino acid ester or amide and NMM dissolved in DMF was added, and the mixture was stirred at −15 ° C. or lower for 30 minutes. Then, it stirred at room temperature for 60 minutes and confirmed completion | finish of reaction using TLC. Thereafter, extraction was performed with AcOEt, acid / base washing was performed to leave an AcOEt layer, and sodium sulfate was added for drying. Sodium sulfate was removed by filtration and then solidified by concentration under reduced pressure to obtain the desired product.

2. Peptide deprotection
4N HCl / DOX was added to the Boc amino acid or peptide and allowed to react at room temperature for 1-2 hours. Thereafter, N ₂ gas was added to drive off excess HCl and solidified to obtain the desired product.

Table 7 shows the yield and yield of each tyrosine derivative synthesized according to 1-2 above.

Each tyrosine derivative shown in Table 7 was confirmed to be the target product by the following TLC, HPLC and ¹ H-NMR.

3. Thin layer chromatography (TLC)
A TLC plate manufactured by Merck (Silica gel 60) was used.
The developing solvent used was C (Chloroform): M (Methanol) = 19: 1 (v / v) or 9: 1 (v / v).

4). High performance liquid chromatography (HPLC)
The column used was 5C ₁₈ -PAQ size 4.6 × 150 mm (Nacalai Tesque). Elution was a water-acetonitrile-0.05% TFA system, and the acetonitrile content was linearly changed from 10 to 90% in 30 to 40 minutes. The flow rate was 1.0 ml / min, and the absorbance at 215 nm was used for detection.
As an example of the measurement result, H-Tyr (Bzl) -OPr (C ₃ ) is shown in FIG. 5A.

5. Nuclear Magnetic Resonance ⁽¹ H-NMR)
A sample for measurement was prepared by dissolving about 7 to 10 mg of the tyrosine derivative synthesized above in about 750 to 1000 μl of CDCl ₃ or DMSO-d6. TMS (Tetramethyl silane) was used as an internal standard.
It was confirmed from the ¹ H-NMR spectrum that the desired product was obtained.
As an example of the measurement result, the spectrum of H-Tyr (Bzl) -OPr (C ₃ ) is shown in FIG. 5B.

6). Measurement of antifungal activity The antifungal activity of each of the synthesized tyrosine derivatives against Valsa ceratosperma Maire was measured.

  1.5% potato dextrose agar (PDA) medium was heated and dissolved, cooled to about 50 ° C., and then spores of apple rot fungus (Valsa ceratosperma Maire) were suspended. The spore suspension was spread in a petri dish and allowed to solidify at room temperature. Samples dissolved in DMSO to 25 mg / ml were used as measurement samples, and 50 μl each was spotted on a paper disk placed on the medium. After culturing at 25 ° C. for 3 days, antifungal activity was observed.

The results are shown in Table 8 and FIG.
The diameter of the blocking circle is the diameter of the blocking circle including the paper disk, "-" is ~ 10 mm, "+" is 10-20 mm, "++" is 20-30 mm, "++" is 30 mm Indicate

As a result, C _{1 to} C ₆ showed clear antifungal activity, but C ₁₂ and C ₁₈ showed no activity. The results of measuring the antifungal activity of C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₁₂ and C ₁₈ are summarized in FIG. As shown in FIG. 7, C ₄ has the highest activity among C _{1 to} C ₆ , and the antifungal activity increases every time the carbon number increases from C _1, and every time the carbon number increases from C ₄ as a boundary. Activity decreased.
In addition, —NH ₂ showed no activity, but —NHCH ₃ showed activity.

The antifungal activity of the tyrosine derivatives examined in this example is highest for C ₄ , and C ₁₂ and C ₁₈ do not show antifungal activity, so that the antifungal activity disappears if the alkyl chain of the ester becomes too long. Is clear. Therefore, the alkyl chain of the C-terminal esters of tyrosine derivatives have a length suitable to exert antifungal activity, to take a stable structure tyrosine derivatives of C ₄ alkyl chain by stacking, activity most It may have increased (FIG. 8). Further, reduction in antifungal activity in C ₁₂ and C ₁₈ is likely to be associated with reduction in water-soluble.

In addition, although —NH ₂ did not show activity, —NHCH ₃ showed activity, and thus the hydrophobicity on the amino acid C-terminal side and antifungal activity may be related.

  As described above, the amino acid derivatives of some embodiments of the present invention are shown to have antifungal activity.

In this example, the following five tyrosine derivatives were synthesized.

The carboxyl group of tyrosine and the amino group of AA (amino acid) form a peptide bond.

1. Synthesis of tyrosine derivatives
H-Tyr (Bzl) -Gly-OH, H-Tyr (Bzl) -β-Ala-OH, H-Tyr (Bzl) -GABA-OH, H-Tyr (Bzl) -Ava-OH, and H-Tyr (Bzl) -Aca-OH was synthesized by the following procedure.

(1) Synthesis of Boc-Tyr (Bzl) -OSu
Boc-Tyr (Bzl) -OH (2.60 g, 7 mmol) and HOSu (0.866 g) were dissolved in DMF (10 m), and the dehydrating agent DCC (1.59 g) was added under ice-cooling. Stir. After completion of the reaction, by-product DCUrea produced from DCC was filtered off using ethyl acetate. The filtrate was concentrated under reduced pressure, solidified with petroleum ether, and collected by filtration.

(2) Synthesis of Boc-Tyr (Bzl) -AA-OH
As an amino acid to be bound to Boc-Tyr (Bzl) -OSu, one of Gly, β-Ala, GABA, Ava and Aca is used, and Boc-Tyr (Bzl) -Gly-OH and Boc-Tyr (Bzl) are used. -β-Ala-OH, Boc-Tyr (Bzl) -GABA-OH, Boc-Tyr (Bzl) -Ava-OH, and Boc-Tyr (Bzl) -Aca-OH were synthesized. Step (2) was all performed on a 1 mmol scale.

  NMM (0.11 mL) and Boc-Tyr (Bzl) -OSu (1 mmol) obtained in (1) were added to a water-dioxane solution of the amino acid to be coupled (1.2 equivalents) under ice-cooling. Stir overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure. A 10% aqueous citric acid solution was added to the residue to adjust the pH to 3, and the target product was extracted with ethyl acetate. The organic layer was washed and dried over sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure, then solidified with petroleum ether, and the N-terminal protected dipeptide was collected by filtration.

(3) Dipeptide de-Boc
The N-terminal protected dipeptide synthesized in (2) was dissolved in 4N HCl / DOX and left at room temperature for 1-2 hours. After completion of the reaction, excess HCl was blown off with nitrogen gas. Dipeptide hydrochloride with Boc group removed after solidification with diethyl ether, namely H-Tyr (Bzl) -Gly-OH · HCl, H-Tyr (Bzl) -β-Ala-OH · HCl, H-Tyr (Bzl) -GABA-OH.HCl, H-Tyr (Bzl) -Ava-OH.HCl, and H-Tyr (Bzl) -Aca-OH.HCl were obtained.

(4) ¹ H-NMR and HPLC chart For each compound obtained in (3), in the same manner as in Example 1, ¹ H-NMR (TMS (trimethylsilane) as internal standard) measurement and HPLC Analyzed with 9 to 13 show the ¹ H-NMR spectrum and HPLC chart of each compound.

In this example, the following three tyrosine derivatives were synthesized.

(1) Synthesis of isobutyl and s-butyl esters of Tyr (Bzl): general synthesis method by cesium salt method In this example, tyrosine was used as an amino acid. Boc amino acid (1 mmol) was dissolved in MeOH (2 ml), water (0.66 ml) and CsCO ₃ (0.5 mmol) were added and stirred. The solution was concentrated under reduced pressure and further dried with a vacuum pump to obtain an amino acid cesium salt. The residue was dissolved in DMF (3 ml), alkyl halide (1 mmol) was added, and the mixture was stirred overnight while protected from light with aluminum foil. After completion of the reaction, precipitated CsBr was removed by filtration, and the target product was extracted from the filtrate with ethyl acetate. The organic layer was washed with water, 4% NaHCO ₃ , and further with water, and then dried over sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain a Boc amino acid ester.
Boc amino acid ester was dissolved in 4N HCl / DOX and allowed to stand at room temperature for 1-2 hours. After completion of the reaction, excess HCl was blown off with nitrogen gas and solidified with diethyl ether to obtain the desired ester hydrochloride.

According to the above general method, using 1-bromo-2-methylpropane and 2-bromobutane as alkyl halides respectively, isobutyl ester of Tyr (Bzl) (H-Tyr (Bzl) -OBu ⁱ · HCl or H-Tyr) (Bzl) isobutyl ester) and was synthesized Tyr (Bzl) in the s- butyl ester (H-Tyr (Bzl) -OBu s · HCl or H-Tyr (Bzl) sec- butyl ester).

(2) Synthesis of t-butyl ester of Tyr (Bzl)
By transesterification using Boc-Tyr (Bzl) -OH, perchloric acid and t-butyl acetate, the t-butyl ester of Tyr (Bzl) (H-Tyr (Bzl) -OBu ^t · HCl or H-Tyr (Bzl) tert-butyl ester) was synthesized. The experimental procedure followed Taschner et al.'S report (Annalen der Chemie., 646, 127 (1961).). In this method, the Boc group is removed during the treatment with perchloric acid.

(3) ¹ H-NMR chart For each compound obtained in (1) and (2), ¹ H-NMR (TMS (trimethylsilane) as internal standard) was measured in the same manner as in Example 1. did. 14 to 16 show ¹ H-NMR spectra of the respective compounds.

  Antifungal activity was measured for the five dipeptides synthesized in Example 4 and the three esters synthesized in Example 5 by the same method as in Example 1. Samples dissolved in DMSO to a concentration of 25 mg / ml were used as measurement samples, and 50 μl each was spotted on a paper disk in the medium.

The photograph which shows the mode of the petri dish at the time of a measurement is shown in FIG. In FIG. 17, “Gly”, “β-Ala”, “GABA”, “Ava” and “Aca” are H-Tyr (Bzl) -Gly-OH, H-Tyr (Bzl) -β-Ala, respectively. -OH, H-Tyr (Bzl) -GABA-OH, H-Tyr (Bzl) -Ava-OH, and H-Tyr (Bzl) -Aca-OH, meaning "isobutyl", "s-butyl" and "t- butyl" is independently an ^{H-Tyr (Bzl) -OBu i} , H-Tyr (Bzl) -OBu s and ^{H-Tyr (Bzl) -OBu t} . H-Tyr (Bzl) -OBu ^t shows data 10 mg / ml and 25 mg / ml.

As shown in FIG. 17, it was confirmed that all eight measured compounds have antifungal activity. In particular, the ester form of Tyr (Bzl), a tyrosine derivative, showed a relatively strong antifungal activity.

Claims (11)

Formula (I)
NHR ¹ —CHR ² —C (O) —R ³ (I)
[Where
R ¹ represents a hydrogen atom;
R ² represents an aryl C _1-6 alkyl group or a 5- to 10-membered heteroaryl C _1-6 alkyl group (wherein the aryl of the aryl C _1-6 alkyl group is a substituent represented by the formula (II)) The heteroaryl of the 5-10 membered heteroaryl C _1-6 alkyl group may have a substituent represented by the formula (II)), or R ¹ and R ² are combined A 5-membered non-aromatic heterocyclic group having a substituent of formula (II) may be formed;
-O-R ^2a (II)
(In the formula, R ^2a has an aryl C _1-6 alkyl group which may have a substituent, an aryl C _1-6 alkyl-OCO- which may have a substituent, and a substituent. An optionally substituted 5-10 membered heteroaryl C _1-6 alkyl group or an optionally substituted 5-10 membered heteroaryl C _1-6 alkyl-OCO-);
R ³ represents —NR ⁴ R ⁵ or —OR ⁶ ;
R ⁴ and R ⁵ are the same or different and each represents a hydrogen atom or a C _1-6 alkyl group (provided that one of R ⁴ and R ⁵ represents a group other than hydrogen);
R ⁶ represents a C _1-11 alkyl group. ]
Or a pharmaceutically acceptable salt thereof, an antifungal agent. Formula (Ib)
NHR ¹ —CHR ² —C (O) —R ^3b (Ib)
[Where
R ¹ represents a hydrogen atom;
R ² represents an aryl C _1-6 alkyl group or a 5- to 10-membered heteroaryl C _1-6 alkyl group (wherein the aryl of the aryl C _1-6 alkyl group is a substituent represented by the formula (II)) The heteroaryl of the 5-10 membered heteroaryl C _1-6 alkyl group may have a substituent represented by the formula (II)), or R ¹ and R ² are combined A 5-membered non-aromatic heterocyclic group having a substituent of formula (II) may be formed;
-O-R ^2a (II)
(In the formula, R ^2a has an aryl C _1-6 alkyl group which may have a substituent, an aryl C _1-6 alkyl-OCO- which may have a substituent, and a substituent. An optionally substituted 5-10 membered heteroaryl C _1-6 alkyl group or an optionally substituted 5-10 membered heteroaryl C _1-6 alkyl-OCO-);
R ^3b represents -NR ^4b R ^5b ;
R ^4b and R ^5b are the same or different and each represents a hydrogen atom or — (CH ₂ ) _m COOR ⁷ (provided that one of R ^4b and R ^5b represents a group other than hydrogen).
(In the formula, m represents an integer of 0 to 12, and R ⁷ represents a hydrogen atom or a C _1-6 alkyl group optionally having a substituent). ]
Or a pharmaceutically acceptable salt thereof, an antifungal agent. R ¹ is a hydrogen atom, R ² is indolyl —CH ₂ —, or a benzyl group having a substituent represented by the formula (II),
In Formula (II), R ^2a may have a substituent, a benzyl group, an optionally substituted benzyl-OCO—, or an optionally substituted pyridyl-CH ₂ —. or have a substituent is a pyridyl -CH ₂ -OCO-, antifungal agent according to claim 1 or 2. R ¹ and R ² are combined to form a pyrrolidinyl group having a substituent represented by the formula (II),
In Formula (II), R ^2a may have a substituent, a benzyl group, an optionally substituted benzyl-OCO—, or an optionally substituted pyridyl-CH ₂ —. or have a substituent is a pyridyl -CH ₂ -OCO-, antifungal agent according to claim 1 or 2. The following compound groups:

An antifungal agent comprising at least one compound selected from: or a pharmaceutically acceptable salt thereof. The following compound groups:

At least one compound selected from: or a pharmaceutically acceptable salt thereof. Formula (IIIb)

[Wherein, -AA ^b -OH represents -NH- (CH ₂ ) _p -COOH, and p represents an integer of 2 to 5. ]
Or a pharmaceutically acceptable salt thereof.   The compound according to claim 6 or 7, or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is a hydrochloride.   A food or drink comprising the compound defined in any one of claims 1 to 5, the compound according to claim 6 or 7, or a pharmaceutically acceptable salt thereof.   A cosmetic comprising the compound according to any one of claims 1 to 5, the compound according to claim 6 or 7, or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising the compound as defined in any one of claims 1 to 5, the compound according to claim 6 or 7, or a pharmaceutically acceptable salt thereof.