JP7181550B2 - Peptides, pharmaceutically acceptable salts thereof, or prodrugs thereof, and uses thereof - Google Patents

Description

One aspect of the present invention relates to peptides or pharmaceutically acceptable salts thereof, or prodrugs thereof, that are selective agonists of the type 1 neuromedin U receptor (NMUR1).

Another aspect of the present invention relates to an activator of type 1 neuromedin U receptor comprising the peptide or a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient.

Another aspect of the invention relates to a method of activating a type 1 neuromedin U receptor comprising contacting a subject with the peptide or a pharmaceutically acceptable salt thereof, or a prodrug thereof.

Another aspect of the present invention is metabolic syndrome, obesity, diabetes, dyslipidemia, cardiovascular disease, or circadian disease, comprising the peptide, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient. The present invention relates to prophylactic and/or therapeutic agents for rhythm disorders.

Another aspect of the present invention is to treat metabolic syndrome, obesity, diabetes, dyslipidemia, circulation, including administering to a patient an effective amount of the peptide or a pharmaceutically acceptable salt thereof, or a prodrug thereof. The present invention relates to methods for preventing and/or treating organ diseases or circadian rhythm disorders.

Neuromedin U (NMU) is a bioactive peptide that has been pointed out to be related to physiological phenomena such as food intake suppression, energy metabolism enhancement, body temperature rise, weight loss, cardiovascular disease, and circadian rhythm, and is an anti-obesity drug. It is expected to be applied as a medicine such as a therapeutic drug for diabetes and the like (Patent Documents 1 and 2).

The seven amidated C-terminal residues of NMU (Phe ⁰ -Leu ¹ -Phe ² -Arg ³ -Pro ⁴ -Arg ⁵ -Asn ⁶ -NH ₂ ) identified in each animal are completely identical in mammals. It is a sequence and is believed to be an important site for activation of type 1 NMU receptors (NMUR1) and type 2 NMU receptors (NMUR2). It is known that NMUR1 is mainly highly expressed in peripheral tissues (mainly intestinal tract, lung, etc.) and NMUR2 is highly expressed in the central (mainly hypothalamic paraventricular nucleus). Development of agonists for these NMUR1 and NMUR2 is being studied (Non-Patent Document 1).

For example, Non-Patent Document 2 describes (2-thienylacetyl)-Trp ¹ -(4-fluorophenylalanine) ² -Arg ³ -Pro ⁴ -Arg ⁵ -Asn ⁶ -(NH ₂ ) (hexa) as an agonist for NMUR1. Peptide 5d) is disclosed.

International Publication No. 2006/068326 (corresponding to US Patent Application Publication No. 2008/0124335) WO2007/109135

K. Takayama et al. , "Discovery of Selective Hexapeptide Agonists to Human Neuromedin U Receptors Types 1 and 2", Journal of Medicinal Chemistry (2014) 57, 6583-6593 K. Takayama et al. , "Discovery of Potent Hexapeptide Agonists to Human Neuromedin U Receptor 1 and Identification of Their Serum Metabolites", ACS Medicinal Chemistry 6, 305-7 Letters (2031)

However, conventional NMUR1 agonists still have room for improvement from the viewpoint of NMUR1 selectivity (property showing low agonist activity against NMUR2 and high agonist activity against NMUR1). In particular, conventional NMUR1 agonists are said to exhibit high agonist activity not only for NMUR1 but also for NMUR2 at high concentrations (for example, about 10 nM to 100 nM in the method for evaluating NMUR agonist activity described in Examples). I had a problem.

Accordingly, the present invention has been made in view of the above circumstances, and an object of the present invention is to provide an NMUR1 agonist having excellent NMUR1 selectivity, particularly NMUR1 selectivity at high concentrations.

The present inventors have conducted extensive research to solve the above problems. As a result, the inventors have found that the above problems can be solved by a peptide represented by the following formula (1), a pharmaceutically acceptable salt thereof, or a prodrug thereof, thereby completing the present invention.

In the above formula (1),
X ⁰ is a substituted or unsubstituted Ala residue whose side chain is substituted with a substituent selected from the group consisting of an alicyclic group, an aromatic hydrocarbon group, an aralkyl group and a heterocyclic group, or is absent; can be;
X ¹ is a substituted or unsubstituted Ala residue or an α- is a methylalanine residue;
X ² is an Ala residue whose side chain is substituted with a substituent S ^A represented by the following formula (A) or a substituent S ^B represented by the following formula (B), represented by the following formula (A) α-methylalanine residue, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid residue whose side chain is substituted with a substituent S ^A or a substituent S ^B represented by the following formula (B) , and 1,2,3,4-tetrahydronorharman-3-carboxylic acid residues,

In the above formula (A), Y is a condensed 5-membered ring structure or a condensed 6-membered ring structure which may contain 1 to 3 heteroatoms selected from the group consisting of nitrogen atoms, sulfur atoms and oxygen atoms as ring members. Yes; p is an integer of 0 to 2; each R ^A is independently an alkyl group having 1 to 3 carbon atoms;

In formula (B) above, ^q is an integer of 1 to 5; A group selected from the group consisting of a nitro group, a halogenated alkyl group having 1 to 6 carbon atoms, an aromatic hydrocarbon group having 6 to 24 carbon atoms, a heterocyclic group, and an acyl group; indicate the binding site with the side chain;
X ³ is a basic amino acid residue;
^X4 is a Pro residue or a homoproline residue;
Z ₁ represents a hydrogen atom, a hydrocarbon group, a heterocyclic group, an acyl group, a sulfo group, a carboxyl group, a glyoxyl group, a polyalkylene glycol group, or a carrier; and Z ₂ represents an amino group, a hydrogen atom, a hydroxy group , an alkoxy group, a hydrocarbon group, a polyalkylene glycol group, or a carrier.

ADVANTAGE OF THE INVENTION According to this invention, the NMUR1 agonist excellent in NMUR1 selectivity, especially NMUR1 selectivity in a high concentration can be provided.

FIG. 1A shows the results of evaluating the concentration-dependent agonist activity of the peptide obtained in Synthesis Example 22 and human NMU for NMUR1. FIG. 1B shows the results of evaluating the concentration-dependent agonist activity of the peptide obtained in Synthesis Example 22 and human NMU for NMUR2. FIG. 2 shows the evaluation results of the stability in human serum of the peptides obtained in Synthesis Example 1 (Comparative Example) and Synthesis Example 22. FIG. FIG. 3 shows the evaluation results of the stability of the peptides obtained in Synthesis Examples 22 and 36 in human serum. FIG. 4 shows the evaluation results of the stability of the peptides obtained in Synthesis Examples 16 and 34 in human serum. FIG. 5 shows the results of evaluating the stability of the peptides obtained in Synthesis Examples 22 and 36 in rat serum. FIG. 6 shows the evaluation results of the stability of the peptides obtained in Synthesis Examples 16 and 34 in rat serum. FIG. 7 shows the analysis results of blood kinetics in rats of the peptides obtained in Synthesis Example 22 and Synthesis Example 36. FIG. FIG. 8 shows the evaluation results of the weight gain-suppressing activity of the peptide obtained in Synthesis Example 22 upon subcutaneous administration to mice.

The peptide represented by formula (1) above, a pharmaceutically acceptable salt thereof, or a prodrug thereof can function as an NMUR1 agonist with excellent NMUR1 selectivity, particularly NMUR1 selectivity at high concentrations.

Embodiments of the present invention will be described below. In addition, the present invention is not limited only to the following embodiments.

In this specification, "X to Y" indicating a range means "X or more and Y or less". In addition, unless otherwise specified, operations and measurements of physical properties are performed under the conditions of room temperature (20 to 25° C.)/relative humidity of 40 to 50% RH.

In the present specification, "the peptide represented by formula (1) or a pharmaceutically acceptable salt thereof" is also simply referred to as "the peptide according to the present invention". In addition, "type 1 neuromedin U receptor" is also simply referred to as "NMUR1". "Type 2 neuromedin U receptor" is also simply referred to as "NMUR2."

As used herein, "NMUR1 selective agonist" or "selectively activates NMUR1" means that when the agonist activity of each peptide for NMUR1 and NMUR2 is evaluated by the method described in the Examples, the following requirements Means satisfying requirements (1) and requirement (2) at the same time:
Requirement (1) a relative agonist activity to NMUR1 of 30% or more at a concentration of 10 nM, and requirement (2) a relative agonist activity to NMUR2 of 50% or less at a concentration of 100 nM.

From the viewpoint of more strongly activating NMUR1, the relative agonist activity to NMUR1 of the peptide is preferably 50% or more at a concentration of 10 nM, more preferably 30% or more at a concentration of 1 nM. More preferably, it is 50% or more at a concentration and 30% or more at a concentration of 1 nM.

From the viewpoint of even better NMUR1 selectivity, the relative agonist activity of the peptide to NMUR2 is preferably 30% or less at a concentration of 10 nM, and more preferably 30% or less at a concentration of 100 nM.

From the viewpoint of even better NMUR1 selectivity, in one embodiment, "relative agonist activity to NMUR2 at a concentration of 100 nM" relative to "relative agonist activity to NMUR1 at a concentration of 10 nM" (hereinafter, "NMUR1 at a concentration of 10 nM "relative agonist activity to NMUR2 at a concentration of 100 nM" relative to "relative agonist activity to" is also referred to as "NMUR2 at 100 nM/NMUR1 at 10 nM" or "R2/R1 activity") is 0.6 or less A peptide is provided. In another embodiment, peptides are provided wherein the R2/R1 activity value is 0.5 or less. In a further embodiment, peptides are provided wherein the R2/R1 activity value is 0.2 or less. In yet another embodiment, peptides are provided wherein the R2/R1 activity value is 0.1 or less.

"Amino acid residue" in the present invention means a portion corresponding to one unit of amino acids constituting a peptide or protein on a peptide or protein molecule. More specifically, it means a divalent group derived from an α-amino acid represented by the following formula (2) or formula (2'):

However, the above R' is a side chain of an amino acid, for example, a hydrogen atom in the case of a Gly residue, and a methyl group in the case of an Ala residue. The above R″ is a hydrogen atom or a methyl group (that is, when R″ is a methyl group, it becomes an α-methyl amino acid residue).

However, the above Z represents a nitrogen-containing heterocyclic structure, for example, a structure derived from pyrrolidine for a Pro residue, a structure derived from piperidine for a homoproline residue, 1,2,3,4-tetrahydroisoquinoline-3 - structure derived from 1,2,3,4-tetrahydroisoquinoline for carboxylic acid residues, 1,2,3 for 1,2,3,4-tetrahydronorharman-3-carboxylic acid residues, It is a structure derived from 4-tetrahydronorharman.

As used herein, the term "Ala residue with a substituted side chain" refers to an Ala residue in which at least one hydrogen atom of the methyl group represented by R' in the above formula (2) is substituted with a substituent. For example, an Ala residue in which one hydrogen atom of a methyl group represented by R′ in the above formula (2) is replaced with an indolyl group (1H-indol-3-yl group) is synonymous with a Trp residue, and the hydrogen 1 An Ala residue in which an atom is substituted with a phenyl group is synonymous with a Phe residue. Similarly, the term “α-methylalanine residue having a substituted side chain” refers to an α - refers to methylalanine residues. For example, an α-methylalanine residue in which one hydrogen atom of a methyl group represented by R′ in the above formula (2) is substituted with an indolyl group (1H-indol-3-yl group) is synonymous with α-methyltryptophan residue. and the α-methylalanine residue in which one hydrogen atom is substituted with a phenyl group is synonymous with the α-methylphenylalanine residue.

"Amino acid" is derived from natural or unnatural α-amino acid, and if it can have an optically active form, it may be either L-form or D-form, but L-form is preferred. More specifically, "amino acid" refers to Arg, Lys, Asp, Asn, Glu, Gln, His, Pro, Tyr, Trp, Ser, Thr, Gly, Ala, Met, Cys, Phe, Leu, Val, and Ile and their analogues can be exemplified. Examples of the above-mentioned analogues may be derivatives in which the side chains of the above-mentioned 20 amino acids are substituted with arbitrary substituents. alanine, 4-bromophenylalanine), 2-aminobutyric acid, norleucine, norvaline, isovaline, 2-aminobutanoic acid, 2-aminoisobutyric acid (α-methylalanine), homophenylalanine, 2,3-diaminopropionic acid, 2,4- Diaminobutanoic acid, ornithine, 2-hydroxyglycine, homoserine, hydroxylysine, hydroxyproline, 3,4-didehydroproline, homoproline, homocysteine, homoarginine, homomethionine, 1-methyltryptophan, 5-methyltryptophan, α- Methyltryptophan, aspartic acid esters (e.g., aspartic acid-methyl ester, aspartic acid-ethyl ester, aspartic acid-propyl ester, aspartic acid-cyclohexyl ester, aspartic acid-benzyl ester, etc.), glutamic acid esters (glutamic acid-cyclohexyl ester, glutamic acid -ethyl ester, glutamic acid-propyl ester, glutamic acid-methyl ester, glutamic acid-benzyl ester, etc.), formyltryptophan, 2-cyclopentylglycine, 2-cyclohexylglycine, 2-phenylglycine, 2-pyridylglycine, 3-cyclopentylalanine, 3 - cyclohexylalanine, 3-pyridyl-alanine, 3-pyrazolyl-alanine, 3-furanyl-alanine, 3-thienyl-alanine, methoxyphenylalanine, 3-naphthyl-alanine, 3,3-diphenylalanine, benzoylphenylalanine, biphenylalanine ( For example, amino acids such as 4-phenylphenylalanine), 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid and 1,2,3,4-tetrahydronorharman-3-carboxylic acid can be exemplified. Not restricted. In addition, for those having diastereomers having an asymmetric carbon in the side chain, such as Ile and Thr, natural types (e.g., (2R ^* ,3R ^* )-2-amino-3-methylpentanoic acid, and (2R ^* ,3S ^* )-2-amino-3-hydroxybutanoic acid) and unnatural forms (e.g., (2R ^* ,3S ^* )-2-amino-3-methylpentanoic acid, and (2R ^* ,3R ^* )-2-amino-3-hydroxybutanoic acid) can be used without particular distinction. That is, "Ile" is used as a meaning including both (2R ^* ,3R ^* )-2-amino-3-methylpentanoic acid and (2R ^* ,3S ^* )-2-amino-3-methylpentanoic acid, "Thr" is used to include both (2R ^* ,3S ^* )-2-amino-3-hydroxybutanoic acid and (2R ^* ,3R ^* )-2-amino-3-hydroxybutanoic acid. Preferably, natural diastereomers (that is, (2R ^* ,3R ^* )-2-amino-3-methylpentanoic acid for Ile, (2R ^* ,3S ^* )-2-amino-3 for Thr -hydroxybutanoic acid) is used.

Unless otherwise specified, the amino acid sequences described herein are written in the direction from the N-terminus (amino terminus) to the C-terminus (carboxyl terminus) according to convention.

It is known in the art that each amino acid residue can be replaced with an amino acid residue having similar properties based on differences in its side chains (conservative substitutions). For example, the basic amino acids Arg, Lys, His, 2,3-diaminopropionic acid (Dpr), 2,4-diaminobutanoic acid (Dbu), norarginine, homoarginine, 2,4-diaminopentanoic acid, and Ornithine (Orn) can be substituted for each other. Pro and homoPro can be substituted for each other.

As used herein, "pharmaceutically acceptable salt" means a metal salt, ammonium salt, organic acid salt, inorganic acid salt, or organic salt that does not produce undesired physiological effects after administration to a patient or subject. It is a salt with a base or an inorganic base. More specifically, sodium salts, potassium salts, calcium salts, magnesium salts, barium salts, aluminum salts, zinc salts, ammonium salts, methylamine salts, ethylamine salts, aniline salts, dimethylamine salts, diethylamine salts, pyrrolidine salts, Piperidine Salt, Morpholine Salt, Piperazine Salt, Trimethylamine Salt, Triethylamine Salt, Ethanolamine Salt, Diethanolamine Salt, Triethanolamine Salt, Hydrochloride, Hydrobromide, Nitrate, Sulfate, Phosphate, Formate, Acetate , trifluoroacetate, phthalate, fumarate, oxalate, tartrate, maleate, citrate, succinate, malate, methanesulfonate, benzenesulfonate, and p- Examples include, but are not limited to, toluenesulfonate.

A prodrug of the peptide according to the present invention (hereinafter, "the prodrug of the peptide according to the present invention" is also simply referred to as "prodrug") can be oxidized or reduced by gastric acid or enzymes under physiological conditions in vivo. , refers to a peptide that undergoes hydrolysis or the like to change into a peptide according to the present invention. These peptides are described, for example, in Bundgard, H.; , Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985, etc., from the peptide of the present invention.

As a prodrug, when the side chain of the amino acid residue of the peptide according to the present invention has a carboxyl group, an ester derivative obtained by reacting the carboxyl group with an alcohol, or a combination of the carboxyl group and an amine. Amide derivatives obtained by the reaction can be exemplified. More specifically, for example, the carboxyl group of the peptide side chain is an ester represented by -COOR (R is an alkyl group having 1 to 20 carbon atoms), or -CONHR or -CONRR' (R and R' are Peptides derivatized to an amide group represented by (C 1-20 alkyl group) are exemplified.

As a prodrug, when the side chain of the amino acid residue of the peptide according to the present invention has a hydroxyl group, an acyloxy derivative obtained by reacting the hydroxyl group with an acid anhydride or the like to acylate can be exemplified. More specific examples include peptides in which the hydroxy group of the peptide side chain is derivatized to an acyloxy group represented by —OCOR (R is an alkyl group having 1 to 20 carbon atoms).

Examples of prodrugs include derivatives obtained by acylating, N-oxidizing, alkylating, or phosphorylating the amino group when the side chain of the amino acid residue of the peptide according to the present invention has an amino group. . More specifically, for example, peptides in which the side chain amino group is derivatized to an amide group represented by —NHCOR (R is an alkyl group having 1 to 20 carbon atoms) or —NHCOCH(NH ₂ )CH ₃ mentioned.

As used herein, a "substituent selected from the group consisting of an alicyclic group, an aromatic hydrocarbon group, an aralkyl group and a heterocyclic group" for an amino acid or amino acid residue refers to the α carbon of the amino acid or amino acid residue. is a side chain substituent attached to the The number of substituents is usually 0-3, preferably 0-2, more preferably 0-1 per amino acid.

The alicyclic group used herein includes a cyclopropyl group, a cyclopropylalkyl group, a cyclobutyl group, a cyclobutylalkyl group, a cyclopentyl group, a cyclopentylalkyl group, a cyclohexyl group, a cyclohexylalkyl group, a cycloheptyl group, a cyclooctyl group, Alicyclic groups having 3 to 12 carbon atoms such as cyclononyl, cyclodecyl, norbornyl, isobornyl and adamantyl groups can be exemplified, preferably cyclohexyl.

The aromatic hydrocarbon group in the present specification is a group derived from an aromatic hydrocarbon having a monocyclic or condensed ring structure, more specifically, a phenyl group, a naphthyl group, a tolyl group, a phenanthryl group, and the like. , and aromatic hydrocarbon groups having 6 to 20 carbon atoms, preferably phenyl or naphthyl.

The aralkyl group in the present specification is a group in which an alkyl group having 1 to 4 carbon atoms is substituted with the above aromatic hydrocarbon group having 6 to 20 carbon atoms, and more specifically, a benzyl group and a phenethyl group. , a benzhydryl group, and other aralkyl groups having 7 to 24 carbon atoms.

The heterocyclic group in the present specification is a substituent having a monocyclic, condensed bicyclic or condensed tricyclic structure containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur atoms in the ring. More specifically, pyrrolidinyl group, pyrrolyl group, piperidyl group, pyridyl group, imidazolyl group, pyrazolyl group, oxazolyl group, thiazolyl group, morpholyl group, indolyl group, isoindolyl group, benzimidazolyl group, quinolyl group, carbazolyl group, oxetanyl group, thietanyl group, tetrahydrofuranyl group, tetrahydrothienyl group, furanyl group, thienyl group, tetrahydropyranyl group, tetrahydrothiopyranyl group, benzofuranyl group, benzothiophenyl group, isobenzofuranyl group, isobenzothio Examples include a phenyl group and the like, preferably an indolyl group, a thienyl group, or a benzothiophenyl group.

Substituents for these amino acids or amino acid residues are linear or branched alkyl groups having 1 to 6 carbon atoms, linear or branched alkoxy groups having 1 to 6 carbon atoms, hydroxy groups, amino groups, carboxyl groups. may be substituted by further substituents such as groups, ester groups, carbamoyl groups, amide groups, acyl groups, nitro groups, sulfo groups, sulfonamide groups and/or halogen atoms.

The amino acids or amino acid residues substituted with the above substituents are not particularly limited, but examples include 2-cyclopentylglycine, 2-cyclohexylglycine, 2-phenylglycine, 3-pyridylalanine, 3 - cyclopentylalanine, 3-cyclohexylalanine, 3-pyridyl-alanine, 3-pyrazolyl-alanine, 3-furanyl-alanine, 3-thienyl-alanine (e.g. 3-(2-thienyl)alanine), fluorophenylalanine, chlorophenylalanine , bromophenylalanine, methoxyphenylalanine, ethoxyphenylalanine, 3-biphenylalanine, 3,3-diphenylalanine, 3-(3-benzothienyl)-alanine, 3-indolyl-alanine (ie, Trp), 3-(1-methyl indolyl)alanine (i.e., 1-methyltryptophan), 3-imidazolyl-alanine (i.e., His), 3-(1-methylimidazolyl)alanine (i.e., 1-methylhistidine), 3-naphthyl-alanine (i.e., 3-(1-naphthyl)-alanine, 3-(2-naphthyl)-alanine), 3-benzothiophenyl-alanine, 4-acetyl-phenylalanine, 4-benzoyl-phenylalanine and the like.

<Peptide>
According to one aspect of the present invention, there is provided a peptide represented by the following formula (1) or a pharmaceutically acceptable salt thereof, or a prodrug thereof:

In the above formula (1), X ⁰ has a side chain substituted with a substituent selected from the group consisting of a substituted or unsubstituted alicyclic group, an aromatic hydrocarbon group, an aralkyl group and a heterocyclic group. Ala residue, or missing.

In one embodiment, X ⁰ is a substituted or unsubstituted cyclohexyl, phenyl, naphthyl, pyrrolidinyl, pyrrolyl, piperidyl, pyridyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, morpholyl group , an indolyl group, an isoindolyl group, a benzimidazolyl group, a quinolyl group, a carbazolyl group, an oxetanyl group, a thietanyl group, a tetrahydrofuranyl group, a tetrahydrothienyl group, a furanyl group, a thienyl group, a tetrahydropyranyl group, a tetrahydrothiopyranyl group, a benzofuranyl group, An Ala residue whose side chain is substituted with a substituent selected from the group consisting of a benzothiophenyl group, an isobenzofuranyl group, and an isobenzothiophenyl group, or a defect.

In another embodiment, X ⁰ is a Phe residue, a Trp residue, a His residue, a Tyr residue, a 3-cyclohexylalanine residue, a 3-naphthyl-alanine residue, a 3-pyridyl-alanine residue, a 3 -furanyl-alanine, 3-thienyl-alanine, or lacking.

In another preferred embodiment, X ⁰ is a Phe residue, a 3-thienyl-alanine residue, or absent, in terms of NMUR1 selectivity and agonistic activity against NMUR1.

In another more preferred embodiment in terms of NMUR1 selectivity and agonistic activity against NMUR1, X ⁰ is a 3-thienyl-alanine residue, or absent.

In another more preferred embodiment in terms of NMUR1 selectivity and agonistic activity towards NMUR1, X ⁰ is absent. The length of the peptides according to the invention is 6 or 7 amino acid residues, or 6 amino acid residues if ^X0 is missing.

In the above formula (1), X ¹ has a side chain substituted with a substituent selected from the group consisting of a substituted or unsubstituted alicyclic group, an aromatic hydrocarbon group, an aralkyl group and a heterocyclic group. Ala residue; or an α-methylalanine residue whose side chain is substituted with a substituent selected from the group consisting of a substituted or unsubstituted alicyclic group, an aromatic hydrocarbon group, an aralkyl group and a heterocyclic group is the base. Specifically, for example, X ¹ is a substituted or unsubstituted cyclohexyl group, phenyl group, naphthyl group, pyrrolidinyl group, pyrrolyl group, piperidyl group, pyridyl group, imidazolyl group, pyrazolyl group, oxazolyl group, thiazolyl group, morpholyl group, indolyl group, isoindolyl group, benzimidazolyl group, quinolyl group, carbazolyl group, oxetanyl group, thietanyl group, tetrahydrofuranyl group, tetrahydrothienyl group, furanyl group, thienyl group, tetrahydropyranyl group, tetrahydrothiopyranyl group, benzofuranyl An Ala residue or an α-methylalanine residue, the side chain of which is substituted with a substituent selected from the group consisting of a group consisting of a benzothiophenyl group, an isobenzofuranyl group, and an isobenzothiophenyl group.

In one embodiment, X ¹ is a Trp residue, a Phe residue, a His residue, a Tyr residue, a 3-cyclohexylalanine residue, a 3-pyridyl-alanine residue, a 3-benzothiophenyl-alanine residue, 3-benzofuranyl-alanine, 3,3-diphenylalanine residues, 3-naphthyl-alanine residues, 1-methylhistidine residues, 1-methyltryptophan residues, α-methyltryptophan residues, and 4-benzoyl-phenylalanine residues selected from the group consisting of residues;

In another preferred embodiment, in terms of NMUR1 selectivity and agonistic activity towards NMUR1, X ¹ is a Trp residue, a Phe residue, a His residue, a Tyr residue, a 3-cyclohexylalanine residue, a 3-pyridyl- selected from the group consisting of alanine residues, 3-benzothiophenyl-alanine residues, 3-benzofuranyl-alanine, 3,3-diphenylalanine residues, 3-naphthyl-alanine residues, and 1-methyltryptophan residues; be.

In another more preferred embodiment in terms of NMUR1 selectivity and agonistic activity towards NMUR1, X ¹ is selected from the group consisting of Trp residues, 3-benzothiophenyl-alanine residues, and 1-methyltryptophan residues. be done.

In another more preferred embodiment, in terms of NMUR1 selectivity and agonistic activity towards NMUR1, X ¹ is selected from the group consisting of Trp residues and 1-methyltryptophan residues.

In the above formula (1), X ² is an Ala residue whose side chain is substituted with a substituent S ^A represented by the following formula (A) or a substituent S ^B represented by the following formula (B), α-methylalanine residue, 1,2,3,4-tetrahydroisoquinoline, the side chain of which is substituted with a substituent S ^{A represented by formula (A} ) or a substituent S ^B represented by formula (B) below -3-carboxylic acid residues, and 1,2,3,4-tetrahydronorharman-3-carboxylic acid residues. The present inventors have made intensive studies to obtain an NMUR1 agonist with ^excellent NMUR1 selectivity. found to improve performance. In particular, the adoption of the above-described residue, which is more bulky than 4-fluoro-phenylalanine (eg, CPN-170, which is a comparative example), at the ^X2 position provides excellent NMUR1 selectivity at high concentrations.

In the above formula (A), Y is a condensed 5-membered ring structure or a condensed 6-membered ring structure which may contain 1 to 3 heteroatoms selected from the group consisting of nitrogen atoms, sulfur atoms and oxygen atoms as ring members. Yes; p is an integer of 0 to 2; each R ^A is independently an alkyl group having 1 to 3 carbon atoms; * indicates a binding site to the side chain of an amino acid residue. In one embodiment, in the above formula (A), Y is an aromatic ring or a condensed 5-membered ring structure containing one heteroatom as a ring member selected from the group consisting of a nitrogen atom, a sulfur atom and an oxygen atom; p is 0 or 1; each ^RA is independently an alkyl group having 1 to 3 carbon atoms; * indicates a binding site to the side chain of an amino acid residue.

More specifically, the substituent S ^A represented by formula (A) includes a naphthyl group, an indalenyl group, an indenyl group, an indolyl group, an isoindolyl group, a benzimidazolyl group, a quinolyl group, an isoquinolyl group, a quinoxalinyl group, and a quinazolinyl group. , a cinnolinyl group, a benzofuranyl group, a benzothiophenyl group, an isobenzofuranyl group, an isobenzothiophenyl group, a chromenyl group, a thiochromenyl group, an isochromenyl group, an isothiochromenyl group, and alkyl groups containing these groups having 1 to 3 carbon atoms. A group further substituted with a group, and the like can be exemplified.

In one embodiment, substituent S ^A is selected from the group consisting of S ^A -1 through S ^A -12 below. In the following S ^A -1 to S ^A -12, * indicates the binding site with the side chain of the amino acid residue.

In formula (B) above, ^q is an integer of 1 to 5; A group selected from the group consisting of a nitro group, a halogenated alkyl group having 1 to 6 carbon atoms, an aromatic hydrocarbon group having 6 to 24 carbon atoms, a heterocyclic group, and an acyl group; The binding site with the side chain is indicated. In one embodiment, in the above formula (B), q is an integer of 1 to 5; each R ^B is independently an aromatic hydrocarbon group having 6 to 24 carbon atoms and an aromatic acyl group (eg , a benzoyl group, or a naphthoyl group); * indicates a binding site to the side chain of an amino acid residue. By substituting the side chain of an amino acid residue with a substituent S ^B having a substituent containing an aromatic ring such as a phenyl group or an aromatic acyl group as R ^B , the blood stability is remarkably improved. Become.

From the viewpoint of NMUR1 selectivity and agonist activity against NMUR1, it is preferable that ^RB is at least 4-position in formula (B) above, more preferably q=1 and ^RB is 4-position.

More specifically, the substituent S ^B represented by formula (B) includes a bromophenyl group, an iodophenyl group, a tolyl group, a xylyl group, a mesityl group, an ethylphenyl group, a diethylphenyl group, a propylphenyl group, butylphenyl group, cyclohexylphenyl group, methoxyphenyl group, ethoxyphenyl group, propoxyphenyl group, nitrophenyl group, trifluoromethylphenyl group, trifluoroethylphenyl group, biphenyl group, acetylphenyl group, propionylphenyl group, butyrylphenyl group group, benzoylphenyl group, naphthoylphenyl group, pyridylphenyl group, and the like.

In one embodiment, substituent S ^B is selected from the group consisting of S ^B -1 through S ^B -19 below. In S ^B -1 to S ^B -19 below, * indicates a binding site to the side chain of an amino acid residue.

The peptide according to the present invention exhibits excellent stability in blood. In particular, by using an amino acid residue having a methyl group on the α carbon of ^X2 (that is, an α-methylalanine residue whose side chain is substituted with a substituent S ^A or a substituent S ^B ), stability in blood becomes even better.

From the viewpoint of NMUR1 selectivity and agonistic activity against NMUR1, in a preferred embodiment ^, X ² is an Ala residue whose side chain is substituted with a substituent S ^C , an α- amino acids selected from the group consisting of methylalanine residues, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid residues, and 1,2,3,4-tetrahydronorharman-3-carboxylic acid residues wherein the substituent S ^C is an alkyl group having 1 to 3 carbon atoms, an alkoxy group having 1 to 3 carbon atoms, a nitro group, a halogenated alkyl group having 1 to 3 carbon atoms, a phenyl group and A phenyl group substituted with a substituent selected from the group consisting of a benzoyl group, or a naphthyl group, an indolyl group, or a 1-methylindolyl group. The Ala residue with the side chain substituted with an indolyl group corresponds to the Trp residue, the α-methylalanine residue with the side chain substituted with the indolyl group corresponds to the α-methyltryptophan residue, and 1- Ala residues substituted in the side chain with methylindolyl groups correspond to 1-methyltryptophan residues.

In another more preferred embodiment in terms of NMUR1 selectivity and agonistic activity against NMUR1, X ² is an Ala residue with a side chain substituted with a substituent S ^C , a side chain with a substituent S ^C selected from the group consisting of α-methylalanine residues, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid residues, and 1,2,3,4-tetrahydronorharman-3-carboxylic acid residues wherein said substituent S ^C is the group consisting of tolyl, methoxyphenyl, nitrophenyl, trifluoromethylphenyl, biphenyl, benzoylphenyl, naphthyl and indolyl is selected from

In another more preferred embodiment in terms of NMUR1 selectivity and agonistic activity towards NMUR1, X ² is a Trp residue, a 3-(2-naphthyl)alanine residue, a 4-phenyl-phenylalanine residue, a 4-benzoyl - phenylalanine residues, α-methyltryptophan residues, and 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid residues; X ¹ is an amino acid residue other than the Trp residue ^X2 is a Trp residue or an α-methyltryptophan residue.

In formula (1) above, ^X3 is a basic amino acid residue. Specific examples of basic amino acid residues include Arg residues, Lys residues, His residues, 2,3-diaminopropionic acid residues, 2,4-diaminobutanoic acid residues, norarginine residues ( 2-amino-4-guanidinobutanoic acid residue), homoarginine residue (2-amino-6-guanidinohexanoic acid residue), 2,4-diaminopentanoic acid residue, and ornithine residue.

In one embodiment, ^X3 is an Arg residue, a Lys residue, a His residue, a 2,3-diaminopropionic acid residue, a 2,4-diaminobutanoic acid residue, a norarginine residue, a homoarginine residue , 2,4-diaminopentanoic acid, and ornithine residues.

In another embodiment, ^X3 is selected from the group consisting of Arg, norarginine, and homoarginine residues.

From the point of view of NMUR1 selectivity and agonistic activity against NMUR1, in one preferred embodiment ^X3 is an Arg residue.

In formula (1) above, ^X4 is a Pro residue or a homoproline residue. Preferably ^X4 is a Pro residue residue.

In the above formula (1), _Z1 is the N-terminal structure of the peptide according to the present invention. Z ₁ represents a hydrogen atom (ie, unmodified), hydrocarbon group, heterocyclic group, acyl group, sulfo group, carboxyl group, glyoxyl group, polyalkylene glycol group, or carrier.

Hydrocarbon groups for Z ₁ include methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, t-butyl group, pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, Chain hydrocarbon groups having 1 to 18 carbon atoms such as undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, and octadecyl; cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; , a cycloheptyl group, a cyclooctyl group, a cyclononyl group, a cyclodecyl group, a norbornyl group, an isobornyl group, and an adamantyl group. More specifically, aromatic hydrocarbon groups having 6 to 20 carbon atoms such as phenyl, naphthyl, tolyl, and phenanthryl groups can be exemplified. These hydrocarbon groups include linear or branched alkyl groups having 1 to 6 carbon atoms, linear or branched alkoxy groups having 1 to 6 carbon atoms, amino groups, carboxyl groups, ester groups, carbamoyl groups, amide may be substituted by further substituents such as groups, nitro groups, sulfo groups, sulfonamide groups and/or halogen atoms.

The heterocyclic group for Z ₁ is the above-mentioned substituted monocyclic, condensed bicyclic or condensed tricyclic structure containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur atoms in the ring. group can be exemplified. The heterocyclic group is a linear or branched alkyl group having 1 to 6 carbon atoms, a linear or branched alkoxy group having 1 to 6 carbon atoms, an amino group, a carboxyl group, an ester group, a carbamoyl group, an amide group, It may be substituted by further substituents such as nitro groups, sulfo groups, sulfonamide groups and/or halogens.

The acyl group for Z ₁ includes, for example, aliphatic acyl groups having 1 to 12 carbon atoms (e.g., formyl group, acetyl group, propionyl group, butyryl group, isobutyryl group, valeryl group, pivaloyl group, hexanoyl group, heptanoyl group, octanoyl group, nonanoyl group, decanoyl group, lauroyl group, etc.), vitamins having an acyl group, nucleic acid bases having an acyl group, or groups represented by the following formula (3).

However, in the above formula (3), R ₁ is an alicyclic group having 3 to 12 carbon atoms (e.g., cyclopropyl group, cyclobutyl group, cyclopentyl group, cyclohexyl group, cycloheptyl group, cyclooctyl group, cyclononyl group, cyclodecyl group, norbornyl group, isobornyl group, and adamantyl group), aromatic hydrocarbon groups having 6 to 20 carbon atoms (e.g., phenyl group, naphthyl group, phenanthryl group, etc.), and heterocyclic groups (e.g., pyrrolidinyl group, pyrrolyl group, piperidyl group, pyridyl group, imidazolyl group, imidazolinyl group, pyrazolyl group, oxazolyl group, thiazolyl group, morpholyl group, indolyl group, benzimidazolyl group, quinolyl group, carbazolyl group, oxetanyl group, thietanyl group, tetrahydrofuranyl group, tetrahydro thienyl group, furanyl group, thienyl group, tetrahydropyranyl group, tetrahydrothiopyranyl group, etc.), and the cyclic group may further have a substituent. . R ₂ is a single bond, an alkylene group having 1 to 11 carbon atoms, an oxyalkylene group having 1 to 10 carbon atoms, or an alkyleneoxy group having 1 to 10 carbon atoms. Substituents for R ₁ include linear or branched alkyl groups having 1 to 6 carbon atoms, amino groups, carboxyl groups, ester groups, carbamoyl groups, amide groups, nitro groups, sulfo groups, sulfonamide groups, and halogens. Atoms (eg, fluorine atom, chlorine atom, bromine atom, iodine atom) and the like can be exemplified.

Examples of vitamins having an acyl group include nicotinic acid, pantothenic acid, biotin, pteroylglutamic acid (folic acid), orotic acid, fluoroorotic acid, α-lipoic acid, pyridoxic acid, biocytin, pteroic acid, and 10-formylpteroin. acid, 7,8-dihydrofolic acid, homopteroic acid, pterin-6-carboxylic acid, dihydrolipoic acid, hydroorotic acid and the like.

A nucleic acid base derivative having an acyl group refers to a base component constituting a nucleotide and a derivative thereof, and preferably includes pyrimidine derivatives such as 5-carboxymethyluracil, 5-carboxythiouracil and the like.

The polyalkylene glycol group of Z ₁ is an ester bond, an amine (--NH--), an acyl group (e.g., an acyl group having 1 to 12 carbon atoms), etc., or a combination thereof, to polyethylene glycol, polypropylene glycol, etc. It is a structure in which polyalkylene glycols are linked. Preferably, the polyalkylene glycol group is a polyethylene glycol group. The number of carbon atoms in the polyethylene glycol group is, for example, 2 to 20 (that is, it has a structural unit represented by -(C ₂ H ₄ O) _n -, where n=1 to 10), preferably 4 to 16. (That is, it has a structural unit represented by -(C ₂ H ₄ O) _n -, where n=2 to 8). The end of the polyalkylene glycol group opposite to the side linked to the N-terminus of the peptide is an alkyl group having 1 to 6 carbon atoms (e.g., methyl group, ethyl group, propyl group, isopropyl group, cyclopropyl group, butyl group, isobutyl group, sec-butyl group, tert-butyl group, amyl group, isoamyl group, tert-amyl group, hexyl group). may

As carriers for _Z1 , molecules commonly used as carriers for peptides can be used, such as streptavidin, albumin, ovalbumin, dextran, pullulan, ficoll, polyvinyl alcohol, styrene-maleic anhydride copolymer. , amylose, amylopectin, cyclodextrin and the like.

In a preferred embodiment, Z ₁ is a hydrogen atom, an aliphatic acyl group having 1 to 12 carbon atoms, or a group represented by the above formula (3); provided that in formula (3), R ₁ is a substituted or unsubstituted thienyl, furanyl, pyrrolidinyl, pyrrolyl, imidazolyl, imidazolinyl, pyrazolyl, oxazolyl, thiazolyl, cyclopentyl, cyclohexyl, phenyl, or pyridyl groups (for example, 2 -pyridyl group, 3-pyridyl group, or 4-pyridyl group); R ₂ is a single bond, an alkylene group having 1 to 11 carbon atoms, an oxyalkylene group having 1 to 10 carbon atoms, or 1 to 10 carbon atoms is an alkyleneoxy group of

From the viewpoint of NMUR1 selectivity and agonist activity against NMUR1, in a more preferred embodiment, Z ₁ is a hydrogen atom, an aliphatic acyl group having 1 to 6 carbon atoms, or a group represented by the above formula (3). provided that in formula (3), R ₁ is a substituted or unsubstituted thienyl group, furanyl group, pyrrolidinyl group, pyrrolyl group, imidazolyl group, imidazolinyl group, pyrazolyl group, oxazolyl group, thiazolyl group, cyclopentyl group, cyclo a hexyl group, a phenyl group, a 3-pyridyl group, or a 4-pyridyl group; R ₂ is a single bond, an alkylene group having 1 to 5 carbon atoms, an oxyalkylene group having 1 to 4 carbon atoms, or 1 carbon atom; ∼4 alkyleneoxy groups.

From the viewpoint of NMUR1 selectivity and agonist activity against NMUR1, in a more preferred embodiment, Z ₁ is a hydrogen atom, an aliphatic acyl group having 2 to 6 carbon atoms, or a group represented by the above formula (3). with the proviso that in formula (3), R ₁ is a substituted or unsubstituted thienyl group, furanyl group, pyrrolyl group, imidazolyl group, 3-pyridyl group, or 4-pyridyl group; R ₂ is a single bond; , or an alkylene group having 1 to 5 carbon atoms.

In formula (1) above, Z ₂ represents an amino group, a hydrogen atom (ie, unmodified), a hydroxy group, an alkoxy group, a hydrocarbon group, a polyalkylene glycol group, or a carrier.

Specifically, the alkoxy group, hydrocarbon group, polyalkylene glycol group, and carrier of Z ₂ can be those mentioned above for Z ₁ .

Z ₂ is preferably an amino group or a hydrogen atom, more preferably an amino group, from the viewpoint of NMUR1 agonist _activity .

In one embodiment, in the above formula (1),
X ⁰ is a Phe residue, Trp residue, His residue, Tyr residue, 3-cyclohexylalanine residue, 3-naphthyl-alanine residue, 3-pyridyl-alanine residue, 3-furanyl-alanine, 3 - thienyl-alanine, or is deficient;
X ¹ is Trp residue, Phe residue, His residue, Tyr residue, 3-cyclohexylalanine residue, 3-pyridylalanine residue, 3-benzothiophenylalanine residue, 3-benzofuranylalanine, 3 , 3-diphenylalanine residues, 3-naphthylalanine residues, 1-methylhistidine residues, 1-methyltryptophan residues, α-methyltryptophan residues, and 4-benzoyl-phenylalanine residues. ;
X ² is an Ala residue whose side chain is substituted with a substituent S ^A represented by the above formula (A) or a substituent S ^B represented by the above formula (B), represented by the above formula (A) α-methylalanine residue, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid residue whose side chain is substituted with a substituent S ^A or a substituent S ^B represented by the above formula (B) , and 1,2,3,4-tetrahydronorharman-3-carboxylic acid residues; wherein said substituent S ^A is a naphthyl group, an indalenyl group, an indenyl group, indolyl group, isoindolyl group, benzimidazolyl group, quinolyl group, isoquinolyl group, quinoxalinyl group, quinazolinyl group, cinnolinyl group, benzofuranyl group, benzothiophenyl group, isobenzofuranyl group, isobenzothiophenyl group, chromenyl group, thiochromenyl group group, isochromenyl group, isothiochromenyl group, and groups in which these functional groups are further substituted with alkyl groups having 1 to 3 carbon atoms; the substituent S ^B is bromophenyl group, iodophenyl group, tolyl group, xylyl group, mesityl group, ethylphenyl group, diethylphenyl group, propylphenyl group, butylphenyl group, cyclohexylphenyl group, methoxyphenyl group, ethoxyphenyl group, propoxyphenyl group, nitrophenyl group, trifluoromethyl selected from the group consisting of a phenyl group, a trifluoroethylphenyl group, a biphenyl group, an acetylphenyl group, a propionylphenyl group, a butyrylphenyl group, a benzoylphenyl group, a naphthoylphenyl group, and a pyridylphenyl group;
X ³ is an Arg residue, Lys residue, His residue, 2,3-diaminopropionic acid residue, 2,4-diaminobutanoic acid residue, norarginine residue, homoarginine residue, 2,4- selected from the group consisting of diaminopentanoic acid, and ornithine residues;
^X4 is a Pro residue or a homoproline residue;
Z ₁ is a hydrogen atom, a hydrocarbon group, a heterocyclic group, an acyl group, a sulfo group, a carboxyl group, a glyoxyl group, a polyalkylene glycol group, or a carrier;
_Z2 is an amino group, a hydrogen atom, a hydroxy group, an alkoxy group, a hydrocarbon group, a polyalkylene glycol group, or a carrier.

In another preferred embodiment, in the above formula (1),
X ⁰ is a Phe residue, a 3-thienyl-alanine residue, or missing;
X ¹ is selected from the group consisting of Trp residues, 3-benzothiophenyl-alanine residues, and 1-methyltryptophan residues;
X ² is an Ala residue ^whose side chain is substituted with a substituent selected from the group consisting of S ^A -1 to S ^A -12 and S ^B -1 to S ^B -19; ~ S ^A -12 and an α-methylalanine residue, 1,2,3,4-tetrahydroisoquinoline-, the side chain of which is substituted with a substituent selected from the group consisting of S ^B -1 to S ^B -19 above. X 3 is an amino acid residue selected from the group consisting of 3-carboxylic acid residues and 1,2,3,4-tetrahydronorharman-3-carboxylic acid residues, and X ³ is an Arg residue, a norarginine residue , and homoarginine residues;
^X4 is a Pro residue or a homoproline residue;
Z ₁ is a hydrogen atom, an aliphatic acyl group having 1 to 12 carbon atoms, or a group represented by the above formula (3); provided that in formula (3), R ₁ is a substituted or unsubstituted thienyl, furanyl, pyrrolidinyl, pyrrolyl, imidazolyl, imidazolinyl, pyrazolyl, oxazolyl, thiazolyl, cyclopentyl, cyclohexyl, phenyl, or pyridyl; _R2 is a single bond; , an alkylene group having 1 to 11 carbon atoms, an oxyalkylene group having 1 to 10 carbon atoms, or an alkyleneoxy group having 1 to 10 carbon atoms;
_Z2 is an amino group or a hydrogen atom.

In another particularly preferred embodiment, in the above formula (1),
X ⁰ is missing;
X ¹ is selected from the group consisting of a Trp residue and a 1-methyltryptophan residue;
^X2 is a Trp residue, a 3-(2-naphthyl)alanine residue, a 4-phenyl-phenylalanine residue, a 4-benzoyl-phenylalanine residue, an α-methyltryptophan residue, and a 1,2,3,4 - tetrahydroisoquinoline-3-carboxylic acid residues; provided that when X ¹ is a 1-methyltryptophan residue, X ² is a Trp residue or an α-methyltryptophan residue;
X ³ is an Arg residue;
X ⁴ is a Pro residue;
Z ₁ is a hydrogen atom, an aliphatic acyl group having 2 to 6 carbon atoms, or a group represented by the above formula (3); provided that in formula (3), R ₁ is a substituted or unsubstituted a thienyl group, a furanyl group, a pyrrolyl group, an imidazolyl group, a 3-pyridyl group, or a 4-pyridyl group;
R ₂ is a single bond or an alkylene group having 1 to 5 carbon atoms;
_Z2 is an amino group.

From the viewpoint of NMUR1 selectivity and agonist activity against NMUR1, in a preferred embodiment, the peptide represented by formula (1) is a peptide satisfying either (i) or (ii) below or a pharmaceutically acceptable Provided are salts, or prodrugs thereof, of:
(i) X ⁰ -X ¹ -X ² -X ³ -X ⁴ -Arg-Asn in the above formula (1) consists of an amino acid sequence selected from the group consisting of SEQ ID NOS: 1 to 21; (ii) SEQ ID NOS: In the amino acid sequence selected from the group consisting of 1 to 21, any one of the amino acid residues at positions corresponding to X ⁰ to X ⁴ are the amino acid residues defined herein for X ⁰ to X ⁴ respectively and selectively activates NMUR1.

The meanings of the abbreviations used in this specification are shown below.
Ala (2-thienyl): 3-(2-thienyl)alanine Ala (1-Naph): 3-(1-naphthyl)alanine Ala (2-Naph): 3-(2-naphthyl)alanine Ala (3-Bzt ): 3-benzothiophenyl-alanine (i.e. 3-(1-benzothiophen-3-yl)-alanine)
Dph: 3,3-diphenylalanine Phe (4-benzoyl): 4-benzoyl-phenylalanine Phe (4-Br): 4-bromo-phenylalanine Phe (4-CF ₃ ): 4-trifluoromethyl-phenylalanine Phe (4 -NO ₂ ): 4-nitro-phenylalanine Phe (4-OMe): 4-methoxy-phenylalanine Phe (4-Ph): 4-phenyl-phenylalanine Trp (1-Me): 1-methyltryptophan (α-Me) Trp: α-methyltryptophan Tic: 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid Tpi: 1,2,3,4-tetrahydronorharman-3-carboxylic acid.

In the peptide of (ii) above, “substituted by amino acid residues defined for X ⁰ to X ⁴ respectively” means that any one of X ⁰ to X ⁴ in the amino acid sequences of SEQ ID NOs: 1 to 21 It means that the amino acid residues corresponding to one position are replaced by the amino acid residues described above for each of X ⁰ to X ⁴ .

In one embodiment, the amino acid sequence in the peptide of (ii) above is represented by any one of SEQ ID NOS:26-38.

From the viewpoint of NMUR1 selectivity and agonist activity against NMUR1, in one preferred embodiment, the amino acid sequence in the peptide of (ii) above is represented by any one of the group consisting of SEQ ID NOs: 26-30 and 35-38. .

In one embodiment, the peptide according to the present invention is represented by any one of formulas (1-1) to (1-185) below.

In a more preferred embodiment, the peptide according to the present invention is represented by the above formulas (1-1), (1-3), (1-8) to (1-10), (1-13), (1-15) ), (1-17), (1-20), (1-22) ~ (1-25), (1-29), (1-30), (1-32) ~ (1-35), (1-37) ~ (1-58), (1-60) ~ (1-73), (1-75) ~ (1-96), (1-98) ~ (1-118), (1 -120) to (1-141), (1-143) to (1-164), and (1-165) to (1-184).

In a further preferred embodiment, the peptide of the present invention is represented by the above formulas (1-8), (1-20), (1-22) to (1-25), (1-29), (1-30) ), (1-44), (1-47), (1-49), (1-50), (1-53), (1-68), (1-82), (1-85), (1-87), (1-88), (1-91), (1-105), (1-108), (1-110), (1-111), (1-114), (1 −150), (1-153), (1-155), (1-156), or (1-159).

<Method for producing peptide>
The peptide according to the present invention can be produced by conventionally known methods including chemical synthesis methods and recombinant techniques. In order to prepare the peptide according to the present invention by chemical synthesis, each amino acid is prepared by a method commonly used in peptide chemistry, for example, "The Peptides" Vol. 1 [by Schroder and Luhke, Academic Press, New York, U.S.A. S. A. (1966)], "Fundamentals and Experiments of Peptide Synthesis" (Nobuo Izumiya et al., Maruzen Co., Ltd., 1985). can be produced by any of Furthermore, either a column method or a batch method can be used.

Peptides according to the present invention may also be produced by recombinant technology using animal cells, insect cells, microorganisms, or the like, for example, by methods described in Current Protocols in Molecular Biology, Chapter 16 below. Peptides can be purified by conventionally known methods after being produced by cultured cells or microorganisms. Peptide purification and isolation methods are known to those skilled in the art, and can be performed, for example, by the techniques described in Current Protocols in Molecular Biology, Chapter 16 (Ausubel et al., John Wiley and Sons, 2006).

Condensation methods for forming peptide bonds include the azide method, acid halide method, acid anhydride method, carbodiimide method, carbodiimide-additive method, active ester method, carbonylimidazole method, redox method, enzymatic method, and Woodward reagent K. , a HATU reagent, a method using a Bop reagent, and the like. Among the methods described above, the solid-phase condensation reaction is mainly performed by the acid anhydride method, the carbodiimide method, and the active ester method.

Furthermore, when the peptide chain is extended by the solid-phase method, the C-terminal amino acid is bound to a support such as a resin that is insoluble in the organic solvent used. As such a resin, a resin in which a functional group is introduced for the purpose of binding the amino acid to the resin, a resin in which a spacer is inserted between the resin and the functional group, or the like can be used depending on the purpose. More specific examples include halomethyl resins such as chloromethyl resins, oxymethyl resins, 4-(oxymethyl)-phenylacetamidomethyl resins, 4-(oxymethyl)-phenoxymethyl resins, Rink amide resins, and the like. can be done. Prior to carrying out these condensation reactions, protective means such as carboxyl groups, amino groups, hydroxyl groups, amidino groups and the like, which do not participate in the condensation reactions, can be applied by a generally known means. Conversely, carboxyl groups and amino groups that directly participate in the condensation reaction can also be activated.

Protective groups used for protecting functional groups that do not participate in the condensation reaction of each unit include protective groups commonly used in organic chemistry, such as "Protective Groups in Organic Synthesis" by Greene, John Wiley & Sons, Inc. ( 1981)), etc. More specifically, protective groups for carboxyl and amino groups include, for example, various methyl esters, ethyl esters, and benzyl esters. , p-nitrobenzyl ester, t-butyl ester, cyclohexyl ester, trityl group (Trt group), benzyloxycarbonyl group, t-butoxycarbonyl group (Boc group), isobornyloxycarbonyl group, 9-fluorenylmethoxy Carbonyl group (Fmoc group), 2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-5-yl-sulfonyl group (Pbf group) and the like can be mentioned.

Activated carboxyl groups include, for example, acid anhydrides corresponding to the carboxyl groups; azide; pentafluorophenol, 2,4-dinitrophenol, cyanomethyl alcohol, p-nitrophenol, N-hydroxysuccinic acid active esters with imide, N-hydroxy-5-norbornene-2,3-dicarboximide, N-hydroxyphthalimide, 1-hydroxybenzotriazole, and the like; Phosphate amide corresponding to the amino group can be mentioned as the activated amino group.

Condensation reaction during peptide synthesis is usually carried out in a solvent. Examples of the solvent include chloroform, dichloromethane, ethyl acetate, N,N-dimethylformamide, dimethylsulfoxide, pyridine, dioxane, tetrahydrofuran, N-methylpyrrolidone, water, methanol, and mixtures thereof. . In addition, the condensation reaction can be carried out at a temperature in the range of -30°C to 50°C, as in the usual case.

Furthermore, the type of protective group elimination reaction in the peptide production process can be selected according to the type of protective group used as long as the protective group can be eliminated without affecting the peptide bond. . For example, acid treatment with hydrogen chloride, hydrogen bromide, anhydrous hydrogen fluoride, methanesulfonic acid, trifluoromethanesulfonic acid, trifluoroacetic acid, or mixtures thereof, sodium hydroxide, potassium hydroxide, hydrazine, diethylamine, piperidine, etc. alkali treatment with , sodium treatment in liquid ammonia, reduction with palladium carbon, and silylation treatment with trimethylsilyl triflate, trimethylsilyl bromide, and the like. In addition, in the deprotection reaction by treatment with an acid or a silylating agent, the addition of a cation scavenger such as anisole, phenol, cresol, thioanisole, ethanedithiol is effective in deprotection reaction. From this point of view, it is preferable.

The method for cleaving the peptide of the present invention synthesized by the solid phase method from the solid phase also follows a generally known method. For example, treatment with the acid or silylating agent described above can be mentioned as the cleavage method. The peptide of the present invention thus produced can be subjected to commonly known separation and purification means after completion of the series of reactions described above. For example, extraction, distribution, reprecipitation, recrystallization, solid-phase extraction, column chromatography, etc. can be used to obtain the peptide of the present invention in a higher purity.

The peptide according to the present invention can be modified at its N-terminus and C-terminus by conventionally known methods. For N-terminal modification, when peptides are synthesized by the solid-phase method, after deprotection of the last amino acid residue, thienylacetic acid, 3-cyclohexylpropionic acid, or the like is used to modify the N-terminal with the above-mentioned _Z1 . It can be obtained by installing In the modification of the C-terminus, for example, an amide of a peptide can be obtained by solid-phase synthesis using a Rink amide resin, which is a resin for synthesizing an amide.

Peptides according to the invention may be isolated or purified. “Isolated or purified” means that an operation is performed to remove components other than the target component. The purity of the isolated or purified peptide according to the present invention is usually 50% or more (e.g., 70% or more, 80% or more, 90% or more, 95% or more, 98% or more, 99% or more, 100%). be.

<Application of peptide>
One embodiment of the present invention relates to an activator of type 1 neuromedin U receptor (NMUR1 activator), comprising the peptide of the present invention or a prodrug thereof as an active ingredient. Another embodiment of the invention relates to a method of activating a type 1 neuromedin U receptor comprising contacting a subject with a peptide of the invention or a prodrug thereof. The method can be practiced to selectively activate the type 1 neuromedin U receptor (and its signaling pathway) in vivo or in vitro. The "subject" includes humans and non-human animals, including fish, and various cells and tissues derived from these individuals, wherein these individuals, cells and tissues are genetically engineered with type 1 neuromedin U receptor, for example. It may be one that is artificially expressed by a method or the like. Preferably, the "subject" is mammals such as humans, dogs, cats, mice, rats, hamsters, guinea pigs, horses (including racehorses), cows, pigs, rabbits and sheep, chickens, quails and turkeys. poultry such as poultry, and cells and tissues derived from these individuals, more preferably human or human-derived cells.

Another embodiment of the present invention is the prevention and/or prevention of metabolic syndrome, obesity, diabetes, dyslipidemia, cardiovascular disease, or circadian rhythm disorder, comprising the peptide of the present invention or a prodrug thereof as an active ingredient. or a therapeutic agent (hereinafter referred to as "an activator of type 1 neuromedin U receptor comprising a peptide according to the present invention or a prodrug thereof as an active ingredient" and "an agent comprising a peptide according to the present invention or a prodrug thereof as an active ingredient , metabolic syndrome, obesity, diabetes, dyslipidemia, cardiovascular diseases, or circadian rhythm disorders" are also collectively referred to as "agents according to the present invention"). Another embodiment of the present invention relates to metabolic syndrome, obesity, diabetes, dyslipidemia, cardiovascular disease, comprising administering an effective amount of a peptide of the present invention or a prodrug thereof to a patient in need thereof. It relates to methods of prevention and/or treatment of diseases or circadian rhythm disorders.

As used herein, "metabolic syndrome" is a condition in which risk factors for arteriosclerotic disease and type II diabetes, such as visceral obesity, insulin resistance/hyperglycemia, dyslipidemia, and hypertension, accumulate.

As used herein, "obesity" means the condition of being obese or having an excess amount of body fat or adipose tissue compared to lean mass, including obesity based on simple obesity, symptomatic obesity, Pathological conditions or diseases associated with obesity can be exemplified. Symptomatic obesity includes endocrine obesity (Cushing syndrome, hypothyroidism, insulinoma, obesity type II diabetes, pseudohypoparathyroidism, hypogonadism, etc.), central obesity (hypothalamic obesity, frontal lobe syndrome, Kleine-Levin syndrome, etc.), hereditary obesity (Prader-Willi syndrome, Laurence-Moon-Biedl syndrome, etc.), drug-induced obesity (steroids, phenothiazines, insulin, sulfonylurea (SU) agents, β-blocker-induced obesity, etc.) ) can be exemplified. Pathological conditions or diseases associated with obesity include impaired glucose tolerance, diabetes, dyslipidemia, hypertension, heart failure, hyperuricemia/gout, fatty liver (including non-alchoholic steato-hepatitis), coronary artery disease (myocardial infarction, stenosis). heart disease, etc.), cerebral infarction (cerebral thrombosis, transient ischemic attack, etc.), bone/joint disease (osteoarthritis of the knee, osteoarthritis of the hip, spondylosis osteoarthritis, low back pain, etc.), sleeplessness Respiratory syndrome/Pickwick syndrome, menstrual abnormalities (abnormal menstrual cycle, abnormal menstrual volume and cycle, amenorrhea, abnormal paramenstrual symptoms, etc.) can be exemplified.

As used herein, "diabetes" refers to a syndrome characterized by abnormal glucose metabolism and hyperglycemia due to insufficient blood insulin levels or decreased insulin sensitivity, more specifically type II diabetes, obesity Type diabetes can be exemplified. Diabetes also includes complications of diabetes. The complications include neuropathy, nephropathy, retinopathy, diabetic cardiomyopathy, cataract, macroangiopathy, osteopenia, diabetic hyperosmolar coma, infections (respiratory infections, urinary tract infections, gastrointestinal infections, skin and soft tissue infections, lower extremity infections, etc.), diabetic gangrene, xerostomia, deafness, cerebrovascular disorders, peripheral blood circulation disorders, and the like.

As used herein, “dyslipidemia” refers to blood lipid abnormalities including hypertriglyceridemia, hypercholesterolemia, low HDL cholesterolemia, postprandial hyperlipidemia, and mixed dyslipidemia.

As used herein, "cardiovascular disease" includes arteriosclerosis (e.g., atherosclerosis), hypertension, transient ischemic attack, systolic dysfunction, diastolic dysfunction, aneurysm, aortic dissection, myocardium Ischemia, myocardial infarction, angina pectoris, angina, heart failure, dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, cor pulmonale, arrhythmia, valvular heart disease, endocarditis, pulmonary embolism, venous embolism Heart and vascular disorders, including peripheral vascular disease, and peripheral arterial disease, can be exemplified.

As used herein, the term "circadian rhythm disorder" refers to external factors such as shift work and jet lag, or non-24-hour sleep-wake disorders, delayed sleep phase syndrome and advanced sleep phase syndrome. refers to sleep disorders.

By administering the drug according to the present invention to a subject, effects such as suppression of food intake, acceleration of energy metabolism, elevation of body temperature, weight loss, improvement of glucose tolerance, improvement of lipid metabolism, and normalization of body clock can be achieved. The drug according to the present invention may consist of one or more peptides according to the present invention, or one or more prodrugs thereof, or mixtures thereof. The agent of the present invention is generally a pharmaceutical composition containing one or more selected from the peptides of the present invention and prodrugs thereof, and a pharmaceutically acceptable carrier.

The dosage of the peptide or prodrug thereof according to the present invention can be appropriately selected depending on the subject of administration, administration route, target disease, symptoms, and the like. The dose is generally 5 μg to 100 mg/body weight, preferably 500 μg to 50 mg/body weight, more preferably 1 to 10 mg/body weight per dose. In addition, it is possible to reduce the dosage by devising formulation. The number of administrations per day is also not particularly limited, and can be administered, for example, 1 to 5 times per day. The route of administration is also not particularly limited, for example, oral administration, injection (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, intravertebral, etc.), transdermal administration, mucosal administration (e.g., intranasal, inhalation, enteral , vaginal, sublingual) or other means known in the art can be used.

The peptide according to the present invention has NMUR1-selective agonist activity. In addition, the peptide according to the present invention is chemically stable under physiological conditions, such as in blood, and has good blood kinetics. Therefore, based on favorable blood kinetics, the drug according to the present invention can suppress food intake, enhance energy metabolism, and improve body temperature by administration into the body, specifically, local administration such as intravenous, intranasal, and subcutaneous administration. Because it can exert actions such as weight gain, weight loss, improvement of glucose tolerance, improvement of lipid metabolism, and normalization of the body clock, it is effective for metabolic syndrome, obesity, diabetes, dyslipidemia, cardiovascular disease, or circadian rhythm disorder. A prophylactic and/or therapeutic effect can be achieved.

As used herein, a therapeutically effective amount is an amount effective to produce some desired therapeutic effect, commensurate with a reasonable benefit/risk ratio. Similarly, as used herein, "comprising as an active ingredient" means comprising an amount effective to produce some desired therapeutic effect, commensurate with a reasonable benefit/risk ratio.

As used herein, the term "patient" includes non-human animals including humans and fish, preferably humans, dogs, cats, mice, rats, hamsters, guinea pigs, horses (including racehorses), and bovines. , pigs, rabbits and sheep, and poultry such as chickens, quails and turkeys, more preferably humans.

The above pharmaceutically acceptable carriers are not particularly limited, but excipients such as lactose, sucrose, mannitol, starch, corn starch, crystalline cellulose, light silicic anhydride; silica, talc, calcium stearate, stearic acid Lubricants such as magnesium; Binders such as hydroxypropyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, methyl cellulose, polyvinylpyrrolidone, crystalline cellulose, dextrin, gelatin; Ascorbic acid, sodium sulfite, sodium hydrogen sulfite, tocopherol, etc. chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffering agents such as borates, bicarbonates, Tris-HCl, citrates, phosphates, other organic acids; water for injection, physiological saline Solvents such as water, ethanol, propanol, ethylene glycol, propylene glycol, macrogol, olive oil, corn oil; Surfactants or wetting agents such as ruconium, benzethonium chloride, glyceryl monostearate; tonicity agents such as sodium chloride, potassium chloride, glycerin, glucose, sorbitol, mannitol; benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben. antibacterial agents; coloring agents; flavoring agents and diluents; emulsifiers; salt-forming counterions such as sodium; s Pharmaceutical Sciences, 18th ed., edited by AR Gennaro, Mack Publishing Company, 1990).

The content of the peptide according to the present invention or a prodrug thereof in the drug may be 0.01-100% by weight based on the total drug.

<Embodiment>
Embodiments of the present invention are illustrated below, but the present invention is not limited to the following forms.
(1) A peptide represented by the following formula (1) or a pharmaceutically acceptable salt thereof, or a prodrug thereof:

In the above formula (1),
X ⁰ is a substituted or unsubstituted Ala residue whose side chain is substituted with a substituent selected from the group consisting of an alicyclic group, an aromatic hydrocarbon group, an aralkyl group and a heterocyclic group, or is absent; can be;
X ¹ is a substituted or unsubstituted Ala residue or an α- is a methylalanine residue;
X ² is an Ala residue whose side chain is substituted with a substituent S ^A represented by the following formula (A) or a substituent S ^B represented by the following formula (B), represented by the following formula (A) α-methylalanine residue, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid residue whose side chain is substituted with a substituent S ^A or a substituent S ^B represented by the following formula (B) , and 1,2,3,4-tetrahydronorharman-3-carboxylic acid residues,

In the above formula (A), Y is a condensed 5-membered ring structure or a condensed 6-membered ring structure which may contain 1 to 3 heteroatoms selected from the group consisting of nitrogen atoms, sulfur atoms and oxygen atoms; p is an integer of 0 to 2; each R ^A is independently an alkyl group having 1 to 3 carbon atoms; * indicates a binding site to the side chain of an amino acid residue;

In formula (B) above, ^q is an integer of 1 to 5; A group selected from the group consisting of a nitro group, a halogenated alkyl group having 1 to 6 carbon atoms, an aromatic hydrocarbon group having 6 to 24 carbon atoms, a heterocyclic group, and an acyl group; indicate the binding site with the side chain;
X ³ is a basic amino acid residue;
^X4 is a Pro residue or a homoproline residue;
Z ₁ represents a hydrogen atom, a hydrocarbon group, a heterocyclic group, an acyl group, a sulfo group, a carboxyl group, a glyoxyl group, a polyalkylene glycol group, or a carrier; and Z ₂ represents an amino group, a hydrogen atom, a hydroxy group , an alkoxy group, a hydrocarbon group, a polyalkylene glycol group, or a carrier.
(2) X ² is an Ala residue whose side chain is substituted with a substituent S ^C , an α-methylalanine residue whose side chain is substituted with a substituent S ^C , 1,2,3,4-tetrahydroisoquinoline - an amino acid residue selected from the group consisting of a 3-carboxylic acid residue and a 1,2,3,4-tetrahydronorharman-3-carboxylic acid residue;
At this time, the substituent S ^C is a group consisting of an alkyl group having 1 to 3 carbon atoms, an alkoxy group having 1 to 3 carbon atoms, a nitro group, a halogenated alkyl group having 1 to 3 carbon atoms, a phenyl group and a benzoyl group. The peptide or a pharmaceutically acceptable salt thereof according to (1), which is a phenyl group substituted with a substituent selected from, or a naphthyl group, an indolyl group, or a 1-methylindolyl group, or their prodrugs.
(3) X ² is an Ala residue whose side chain is substituted with a substituent S ^C , an α-methylalanine residue whose side chain is substituted with a substituent S ^C , 1,2,3,4-tetrahydroisoquinoline - an amino acid residue selected from the group consisting of a 3-carboxylic acid residue and a 1,2,3,4-tetrahydronorharman-3-carboxylic acid residue;
At this time, the substituent S ^C is selected from the group consisting of tolyl, methoxyphenyl, nitrophenyl, trifluoromethylphenyl, biphenyl, benzoylphenyl, naphthyl, and indolyl groups (2 ) or a pharmaceutically acceptable salt thereof, or a prodrug thereof.
(4) X ¹ is Trp residue, Phe residue, His residue, Tyr residue, 3-cyclohexylalanine residue, 3-pyridyl-alanine residue, 3-benzothiophenyl-alanine residue, 3- Any one of (1) to (3), which is selected from the group consisting of benzofuranyl-alanine, 3,3-diphenylalanine residues, 3-naphthyl-alanine residues, and 1-methyltryptophan residues. or a pharmaceutically acceptable salt thereof, or a prodrug thereof.
(5) Z ₁ is a hydrogen atom, an aliphatic acyl group having 1 to 6 carbon atoms, or a group represented by the following formula (3), according to any one of (1) to (4) peptides or pharmaceutically acceptable salts thereof, or prodrugs thereof;

However, in the above formula (3), R ₁ is a substituted or unsubstituted thienyl group, furanyl group, pyrrolidinyl group, pyrrolyl group, imidazolyl group, imidazolinyl group, pyrazolyl group, oxazolyl group, thiazolyl group, cyclopentyl group, cyclo a hexyl group, a phenyl group, a 3-pyridyl group, or a 4-pyridyl group; R ₂ is a single bond, an alkylene group having 1 to 5 carbon atoms, an oxyalkylene group having 1 to 4 carbon atoms, or 1 carbon atom; ∼4 alkyleneoxy groups.
(6) The peptide or pharmaceutically acceptable salt thereof according to any one of (1) to (5), wherein X ⁰ is a Phe residue, a 3-thienyl-alanine residue, or lacking, or them prodrug of.
(7) The peptide according to any one of (1) to (6), wherein X ³ is selected from the group consisting of an Arg residue, a norarginine residue, and a homoarginine residue, or a pharmaceutically acceptable peptide thereof salts, or prodrugs thereof.
(8) The peptide according to (1), or a pharmaceutically acceptable salt thereof, or a pro drag:
(i) X ⁰ -X ¹ -X ² -X ³ -X ⁴ -Arg-Asn in the above formula (1) consists of an amino acid sequence selected from the group consisting of SEQ ID NOS: 1 to 21; (ii) SEQ ID NOS: any one of the amino acid residues at positions corresponding to X ⁰ to X ⁴ in the amino acid sequence selected from the group consisting of 1 to 21 is the amino acid residue described herein for X ⁰ to X ⁴ respectively and selectively activates NMUR1.
(9) The peptide according to (1), wherein the peptide represented by formula (1) is represented by any one of formulas (1-1) to (1-185) above, or a pharmaceutically Acceptable salts, or prodrugs thereof.
(10) A type 1 neuromedin U receptor activator comprising the peptide according to any one of (1) to (9) or a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient .
(11) Metabolic syndrome, obesity, diabetes, dyslipidemia, containing the peptide according to any one of (1) to (9) or a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient preventive and/or therapeutic agents for diseases, cardiovascular diseases, or circadian rhythm disorders.
(12) administering an effective amount of the peptide according to any one of (1) to (9) or a pharmaceutically acceptable salt thereof, or a prodrug thereof to a patient in need thereof; A method for the prevention and/or treatment of metabolic syndrome, obesity, diabetes, dyslipidemia, cardiovascular disease, or circadian rhythm disorders, including.

The effects of the present invention will be described using the following examples and comparative examples. However, the technical scope of the present invention is not limited only to the following examples.

<Peptide Synthesis>
[Synthesis Example 1] Synthesis of CPN-171 (2-thienylacetyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-171)
CPN-171 was synthesized by the Fmoc solid-phase peptide synthesis method described below.

75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) was weighed into a reaction vessel, and the resin was allowed to swell in a dimethylformamide (DMF) solution at room temperature (25°C) for 30 minutes. After that, the protective group Fmoc (9-fluorophenylmethoxycarbonyl) group on the resin was removed by reacting in 20% (v/v) piperidine/DMF solution (2.5 mL) at room temperature (25°C) for 20 minutes. Removed. The resin was washed 10 times with DMF (2.5 mL), followed by 34 mg (0.22 mmol, 5 eq.) of 1-hydroxybenzotriazole (HOBt) and 0.034 mL (0.22 mmol) of N,N-diisopropylcarbodiimide (DIPCD). , 5 eq.) was reacted with 131 mg (0.20 mmol, 5 eq.) of Fmoc-Asn(Trt)-OH in DMF at room temperature (25° C.) for 1 hour to introduce the amino acid onto the resin. For condensation of the next amino acid, the Fmoc group on the resin was removed by reacting in 20% (v/v) piperidine/DMF solution (2.5 mL) for 20 minutes. Subsequently, Fmoc-Arg(Pbf)-OH (0.22 mmol, 5 eq.), Fmoc-Pro-OH (0.22 mmol, 5 eq.) and Fmoc-Pro-OH (0.22 mmol, 5 eq.) were prepared sequentially from the C-terminal side in the same manner as for Fmoc-Asn(Trt)-OH. ), Fmoc-Arg(Pbf)-OH (0.22 mmol, 5 eq.), Fmoc-Trp(Boc)-OH (0.22 mmol, 5 eq.), Fmoc-Trp(Boc)-OH (0.22 mmol, 5 eq.) .), 2-thienylacetic acid (0.22 mmol, 5 eq.) was introduced to extend the peptide chain. The resin was washed with DMF (2.5 mL, 6 times) and methanol (2.5 mL, 3 times), diethyl ether (2.5 mL, 3 times) and dried. Trifluoroacetic acid (TFA) in the presence of m-cresol (0.100 mL), thioanisole (0.100 mL), and 1,2-ethanedithiol (0.100 mL) for removal of various side-chain protecting groups and deresinization. ) in 4.0 mL for 2 hours. After removing the resin by filter filtration, TFA was distilled off by nitrogen spraying, and 40 mL of diethyl ether was added to precipitate the crude peptide. The crude peptide was dissolved in 1M acetic acid and purified using high performance liquid chromatography to give a white solid (21.6 mg, 79% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1037.4892, found 1037.4874.

[Synthesis Example 2] Synthesis of CPN-172 (2-thienylacetyl)-Trp-Ala(1-Naph)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-172); Ala(1-Naph) : 3-(1-naphthyl)alanine CPN-172 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.). (25.0 mg, 45% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1048.4940, found 1048.4960.

[Synthesis Example 3] Synthesis of CPN-173 (2-thienylacetyl)-Trp-Ala(2-Naph)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-173); Ala(2-Naph) : 3-(2-naphthyl)alanine CPN-173 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.). (31.3 mg, 56% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1048.4940, found 1048.4915.

[Synthesis Example 4] Synthesis of CPN-180 (2-thienylacetyl)-Trp-Phe(4-Br)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-180); Phe(4-Br) : 4-bromo-phenylalanine CPN-180 was synthesized and purified using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) in the same manner as in Synthesis Example 1 (24. 0 mg, 42% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1076.3888, found 1076.3916.

[Synthesis Example 5] Synthesis of CPN-181 (2-thienylacetyl)-Trp-Phe(4-OMe)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-181); Phe(4-OMe) : 4-Methoxy-phenylalanine CPN-181 was synthesized and purified using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) in the same manner as in Synthesis Example 1 (43. 7 mg, 79% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1028.4889, found 1028.4866.

[Synthesis Example 6] Synthesis of CPN-182 (2-thienylacetyl)-Trp-Phe(4-NO ₂ )-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-182); Phe(4-NO ₂ ): 4-nitro-phenylalanine CPN-182 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) ( 42.7 mg, 76% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1043.4634, found 1043.4620.

[Synthesis Example 7] Synthesis of CPN-183 (2-thienylacetyl)-Trp-Phe(4-CF ₃ )-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-183); Phe(4-CF ₃ ): 4-trifluoromethyl-phenylalanine CPN-183 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.). (37.7 mg, 66% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1066.4657, found 1066.4661.

[Synthesis Example 8] Synthesis of CPN-185 (2-thienylacetyl)-Trp-Phe(4-Ph)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-185); Phe(4-Ph) : 4-phenyl-phenylalanine CPN-185 was synthesized and purified using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) in the same manner as in Synthesis Example 1 (36. 1 mg, 63% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1074.5096, found 1074.5071.

[Synthesis Example 9] Synthesis of CPN-186 (2-thienylacetyl)-Trp-Phe(4-benzoyl)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-186); Phe(4-benzoyl) : 4-benzoyl-phenylalanine CPN-186 was synthesized and purified using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) in the same manner as in Synthesis Example 1 (14. 9 mg, 26% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1102.5045, found 1102.5078.

[Synthesis Example 10] Synthesis of CPN-188 (2-thienylacetyl)-Ala(3-Bzt)-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-188); Ala(3-Bzt) : 3-benzothiophenyl-alanine CPN-188 was synthesized and purified using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) in the same manner as in Synthesis Example 1 ( 43.6 mg, 65% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1054.4462, found 1054.4504.

[Synthesis Example 11] Synthesis of CPN-190 (2-thienylacetyl)-Phe(4-benzoyl)-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-190); Phe(4-benzoyl) : 4-benzoyl-phenylalanine CPN-190 was synthesized and purified using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) in the same manner as in Synthesis Example 1 (22. 8 mg, 33% yield).
HRMS (ES+) calcd for (M ⁺ + H) 1102.5063, found 1102.5045.

[Synthesis Example 12] Synthesis of CPN-191 (2-thienylacetyl)-Dph-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-191); Dph: 3,3-diphenylalanine CPN-191 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Kagaku Kogyo Co., Ltd.) (50.0 mg, yield 66%).
HRMS (ES+) calcd for (M ⁺⁺ H) 1074.5096, found 1074.5071.

[Synthesis Example 13] Synthesis of CPN-217 (2-thienylacetyl)-Trp-Tic-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-217); Tic: 1,2,3,4-tetrahydro Isoquinoline-3-carboxylic acid CPN-217 was synthesized and purified using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) in the same manner as in Synthesis Example 1 (22. 5 mg, 41% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1010.4783, found 1010.4778.

[Synthesis Example 14] Synthesis of CPN-218 (2-thienylacetyl)-Trp-Tpi-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-218); Tpi: 1,2,3,4-tetrahydro Norharman-3-carboxylic acid CPN-218 was synthesized and purified by the same method as in Synthesis Example 1 using Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) 75 mg (0.044 mmol) (11 .8 mg, 21% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 895.5154, found 895.5153.

[Synthesis Example 15] Synthesis of CPN-222 (3-pyridinepropionyl)-Ala(2-Naph)-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-222); Ala(2-Naph) : 3-(2-naphthyl)alanine CPN-222 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.). (39.2 mg, 69% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1057.5484, found 1057.5479.

[Synthesis Example 16] Synthesis of CPN-223 (3-pyridinepropionyl)-Ala(2-Naph)-Phe(4-Ph)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-223); (2-Naph): 3-(2-naphthyl) alanine, Phe (4-Ph): 4-phenyl-phenylalanine CPN-223 is Rink Amide resin (0.58 mmol / g, Watanabe Chemical Industry Co., Ltd.) 75 mg (0 .044 mmol) was synthesized and purified by the same method as in Synthesis Example 1 (22.0 mg, yield 38%).
HRMS (ES+) calcd for (M ⁺⁺ H) 1049.5688, found 1049.5693.

[Synthesis Example 17] Synthesis of CPN-259 Ala(2-thienyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-259); Ala(2-thienyl): 3-(2 -thienyl)alanine, the N-terminus of CPN-259 is unmodified (ie, a hydrogen atom).

CPN-259 was synthesized and purified by the same method as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (31.6 mg, yield 56% ).
HRMS (ES+) calcd for (M ⁺⁺ H) 1066.5157, found 1066.5162.

[Synthesis Example 18] Synthesis of CPN-261 (n-hexanoyl)-Ala(2-thienyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-261); ): 3-(2-thienyl)alanine CPN-261 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.). (35.0 mg, 57% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1164.5889, found 1164.5881.

[Synthesis Example 19] Synthesis of CPN-262 (n-decanoyl)-Ala(2-thienyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-262); ): 3-(2-thienyl)alanine CPN-262 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.). (15.2 mg, 24% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1220.6515, found 1220.6512.

[Synthesis Example 20] Synthesis of CPN-265 (2-thienylacetyl)-Trp(1-Me)-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-265); Trp(1-Me) : 1-Methyltryptophan CPN-265 was synthesized and purified by the same method as in Synthesis Example 1 using Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) 75 mg (0.044 mmol) (14.6 mg , yield 34%).
HRMS (ES+) calcd for (M ⁺⁺ H) 1051.4978, found 1051.5045.

[Synthesis Example 21] Synthesis of CPN-266 (2-Thienylacetyl)-Trp-Trp(1-Me)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-266); Trp(1-Me) : 1-Methyltryptophan CPN-266 was synthesized and purified in the same manner as in Synthesis Example 1 using Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) 75 mg (0.044 mmol) (18.4 mg , yield 40%).
HRMS (ES+) calcd for (M ⁺⁺ H) 1051.4978, found 1051.5070.

[Synthesis Example 22] Synthesis of CPN-267 (2-thienylacetyl)-Trp-(α-Me)Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-267); (α-Me)Trp : α-Methyltryptophan CPN-267 was synthesized and purified in the same manner as in Synthesis Example 1 using Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) 75 mg (0.044 mmol) (24.7 mg , yield 56%).
HRMS (ES+) calcd for (M ⁺⁺ H) 1051.5048, found 1051.5059.

[Synthesis Example 23] Synthesis of CPN-276 (2-thienylacetyl)-Trp(1-Me)-(α-Me)Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-276); Trp (1-Me): 1-methyltryptophan, (α-Me)Trp: α-methyltryptophan CPN-276 uses Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) 75 mg (0.044 mmol). (8.1 mg, 17% yield).
HRMS (ES+) calcd for (M ⁺⁺ H) 1065.5205, found 1065.5211.

[Synthesis Example 24] Synthesis of CPN-277 (2-thiophenecarbonyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-277)
CPN-277 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (13.8 mg, yield 33% ).
HRMS (ES+) calcd for (M ⁺⁺ H) 1023.4735, found 1023.4765.

[Synthesis Example 25] Synthesis of CPN-278 (2-furancarbonyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-278)
CPN-278 was synthesized and purified by the same method as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (16.9 mg, yield 42% ).
HRMS (ES+) calcd for (M ⁺ + H) 1007.4964, found 1007.4966.
[Synthesis Example 26] Synthesis of CPN-279 (Pyrrole-2-carboxyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-279)
CPN-279 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (5.4 mg, yield 13% ).
HRMS (ES+)calcd for (M ⁺⁺ H) 1006.5124, found 1006.5158.

[Synthesis Example 27] Synthesis of CPN-280 (2-Pyridylacetyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-280)
CPN-280 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (35.0 mg, yield 64% ).
HRMS (ES+)calcd for (M ⁺⁺ H) 1032.5280, found 1032.5260.
[Synthesis Example 28] Synthesis of CPN-281 (3-Pyridylacetyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-281)
CPN-281 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (32.7 mg, yield 63% ).
HRMS (ES+) calcd for (M ⁺⁺ H) 1032.5280, found 1032.5275.

[Synthesis Example 29] Synthesis of CPN-282 (4-pyridylacetyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-282)
CPN-282 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (21.4 mg, yield 39% ).
HRMS (ES+) calcd for (M ⁺⁺ H) 1032.5280, found 1032.5286.

[Synthesis Example 30] Synthesis of CPN-283 (1H-imidazoleacetyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-283)
CPN-283 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (27.7 mg, yield 67% ).
HRMS (ES+) calcd for (M ⁺⁺ H) 1021.5233, found 1021.5227.

[Synthesis Example 31] Synthesis of CPN-284 (5-bromo-2-thienylacetyl)-Trp-Trp-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-284)
CPN-284 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (28.5 mg, yield 56% ).
HRMS (ES+) calcd for (M ⁺⁺ H) 1115.3997, found 1115.4026.

[Synthesis Example 32] Synthesis of CPN-285 (3-pyridylpropionyl)-Ala(2-Naph)-(α-Me)Trp-Arg-Pro-Arg-Asn-NH ₂ (CPN-285); -Naph): 2-naphthylalanine, (α-Me) Trp: α-methyltryptophan CPN-285 is Rink Amide resin (0.56 mmol / g, Watanabe Chemical Industry Co., Ltd.) 73 mg (0.041 mmol) Synthesized and purified by the same method as in Synthesis Example 1 (28.2 mg, 53%).
HRMS (ES+) calcd for (M ⁺⁺ H) 1071.5641, found 1071.5662.

[Synthesis Example 33] Synthesis of CPN-298 (2-naphthyloxyacetyl)-Tic-Arg-Pro-Arg-Asn-NH ₂ (CPN-298); Tic: 1,2,3,4-tetrahydroisoquinoline-3 -Carboxylic acid CPN-298 was synthesized and purified by the same method as in Synthesis Example 1 using Rink Amide resin (0.56 mmol / g, Watanabe Chemical Industry Co., Ltd.) 175 mg (0.098 mmol) (51.2 mg, 47%).
HRMS (ES+) calcd for (M ⁺⁺ H) 884.4531, found 884.4526.

[Synthesis Example 34] Synthesis of CPN-124 (Comparative Example)
(3-pyridinepropionyl)-Ala(2-Naph)-Phe-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-124); Ala(2-Naph): 3-(2-naphthyl)alanine CPN -124 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (54.5 mg, yield 95%). .
HRMS (ES+) calcd for (M ⁺⁺ H) 1018.5375, found 1018.5395.

[Synthesis Example 35] Synthesis of CPN-125 (Comparative Example)
(2-Thienylacetyl)-Trp-Phe-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-125)
CPN-125 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (39.1 mg, yield 69% ).
HRMS (ES+) calcd for (M ⁺⁺ H) 998.4783, found 998.4792.

[Synthesis Example 36] Synthesis of CPN-170 (Comparative Example)
(2-Thienylacetyl)-Trp-Phe(4-F)-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-170, hexapeptide 5d); Phe(4-F): 4-fluoro-phenylalanine CPN-170 was synthesized and purified in the same manner as in Synthesis Example 1 using 75 mg (0.044 mmol) of Rink Amide resin (0.58 mmol/g, Watanabe Chemical Industry Co., Ltd.) (32.0 mg, yield 59% ).
HRMS (ES+) calcd for (M ⁺⁺ H) 1016.4689, found 1016.4673.

[Synthesis Example 37] Synthesis of CPN-270 (Comparative Example)
(3-pyridinepropionyl)-Ala(2-Naph)-(α-Me)Phe-Arg-Pro-Arg-Asn-(NH ₂ ) (CPN-270); Ala(2-Naph): 3-(2 - Naphthyl) alanine, (α-Me) Phe: α-methylphenylalanine CPN-270 is Rink Amide resin (0.58 mmol / g, Watanabe Chemical Industry Co., Ltd.) 75 mg (0.044 mmol) in the same manner as in Synthesis Example 1 (33.2 mg, yield 76%).
HRMS (ES+) calcd for (M ⁺⁺ H) 1032.5532, found 1032.5525.

[Synthesis Example 38] Synthesis of CPN-111 (Comparative Example)
(2-Naphthyloxyacetyl)-Phe-Arg-Pro-Arg-Asn-NH ₂ (CPN-111)
CPN-111 was synthesized and purified in the same manner as in Synthesis Example 1 using 80 mg (0.044 mmol) of Rink Amide resin (0.55 mmol/g, Watanabe Kagaku Kogyo Co., Ltd.) (46.6 mg, 96%).
HRMS (ES+) calcd for (M ⁺⁺ H) 872.4531, found 872.4531.

[Synthesis Example 39] Synthesis of hNMU (Comparative Example)
Phe-Arg-Val-Asp-Glu-Glu-Phe-Gln-Ser-Pro-Phe-Ala-Ser-Gln-Ser-Arg-Gly-Tyr-Phe-Leu-Phe-Arg-Pro-Arg-Asn- ( _NH2 ) (hNMU)
hNMU was synthesized and purified by the same method as in Synthesis Example 1 using 100 mg (0.055 mmol) of Rink Amide resin (0.55 mmol/g, Watanabe Chemical Industry Co., Ltd.) (92.8 mg, yield 48%).
HRMS (ES+) calcd for (M ⁺⁺ H) 3079.5291, found 3079.5210.

<NMUR agonist activity evaluation>
Agonist activity evaluation of each peptide was carried out by the following method. Table 8 shows the results (relative agonist activity (%) when the agonist activity of human neuromedin U (hNMU) at 1000 nM is taken as 100 (%)). Also, the activity evaluation results for CPN-267 and hNMU are shown in FIGS. 1A and 1B.

(1) Cell culture Chinese hamster ovary-derived CHO cells stably expressing human neuromedin U receptors (NMUR1 and NMUR2), respectively, were prepared using 10% (v/v) calf serum (FCS) containing 1 mg/mL G418. ) were cultured in a 37° C., 5% (v/v) CO ₂ incubator using α-MEM-nucleoside (GIBCO) medium.

(2) In vitro agonist activity evaluation CHO cells were added to 96 well black-walled plate with clear bottom (Iwaki) at 2.0 × 10 ⁴ cells per well (150 µL of DMEM (10% (v/v) fetal bovine serum (FBS)) and cultured for 18 hours.On the next day, assay buffer (Hanks Balanced Salt Solution (HBSS), 10 mM HEPES, 2.5 mM probenecid, 1% ( 100 μL of intracellular calcium concentration fluorescent indicator Fluo-4 AM (Promega) solution prepared by v/v) FCS, pH 7.4) was added to each well.After culturing the cells at 37° C. for 40 minutes, assay buffer After washing four times with , the plate was set in a fluorometric imaging plate reader (Molecular Devices).The peptide was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 20 mM as a stock solution and stored at 4° C. Add to the cells. Stock solutions of peptides were prepared with assay buffer supplemented with 0.05% (v/v) bovine serum albumin (BSA) and 0.001% (v/v) Triton® X-100 1 hour before. The peptide was adjusted to an arbitrary final concentration, and fluorescence due to intracellular calcium mobilization activity (agonist activity) of the peptide was measured using a fluorometric imaging plate reader.

The peptide according to the comparative example had low NMUR1 selectivity. The comparative peptide CPN-170 (Non-Patent Document 2) had low NMUR1 selectivity and did not exhibit full agonist activity even at high concentrations (that is, it was a partial agonist).

On the other hand, as shown in Table 8, the peptides according to the present invention exhibited NMUR1-selective agonist activity. Also, as shown in FIG. 1, the peptide CPN-267 according to the present invention exhibited NMUR1-selective full agonist activity. Among the peptides according to the present invention, CPN-277, CPN-278, CPN-282, and CPN-283 exhibited full agonist activity (NMUR1 relative activity: 93% or more) at 100 nM, similar to CPN-267.

<Evaluation of stability in human serum>
(CPN-171, CPN-267)
In order to verify the stability of CPN-171 (Synthesis Example 1) and CPN-267 (Synthesis Example 22) in human serum, they were evaluated by the following method.

A 20 mM peptide stock solution was diluted once to 1 mM in RPMI1640 medium. 20 μL of this peptide dilution was then added to 380 μL of human serum (Sigma) dilution (100 μL of human serum diluted by adding 280 μL of RPMI 1640 medium) to a final concentration of 50 μM ( 25% final serum concentration). After incubating at 37° C. for an arbitrary time (10 minutes), peptide fractions were collected using a Sep-pak plus C18 (Waters) solid-phase cartridge and subjected to reversed-phase HPLC (column: Chromolith (registered trademark) Performance RP-18e 100 -4.6 mm; gradient: %B 15-55, 10 min [A: H ₂ O-0.1% (v/v) TFA; B: MeCN]; flow rate: 2.5 mL/min; measurement wavelength: 220 nm) The residual rate (%) was analyzed. The residual rate (%) represents the amount of peptide in the solution after incubation when the amount of peptide added to serum before incubation is taken as 100%.

The results are shown in FIG. About 60% of the added amount of CPN-171 remained after 10 minutes of incubation. On the other hand, more than 90% of CPN-267 remained even after 10 minutes of incubation. From the above results, although both CPN-171 and CPN-267 exhibit excellent blood stability, the introduction of a methyl group to the α-carbon of ^X2 results in even better blood stability. I know it will be.

(CPN-170, CPN-267)
Stability of CPN-170 (Synthetic Example 36) and CPN-267 in human serum was measured over time according to the same method as above. The results are shown in FIG. More than 40% of CPN-267 remained after 180 minutes. On the other hand, CPN-170, which is a comparative example, had a residual rate of 25% or less after 10 minutes and about 5% after 20 minutes.

(CPN-124, CPN-223)
The stability of CPN-124 (Synthesis Example 34) and CPN-223 (Synthesis Example 16) in human serum was measured over time according to the same method as above. The results are shown in FIG. More than 80% of CPN-223 remained after 180 minutes. From this result, it was found that the stability in blood is further improved by using a substituent containing an aromatic ring such as a phenyl group as the substituent S ^B of the side chain of the amino acid residue of ^X2 . I understand. On the other hand, CPN-124, which is a comparative example, had a residual rate of about 30% after 20 minutes and about 7% after 40 minutes.

<Evaluation of stability in rat serum>
(CPN-170, CPN-267)
The stability of CPN-170 and CPN-267 in rat serum was evaluated by the same method except that rat serum (Sigma) was used instead of human serum in the above "Evaluation of stability in human serum".

The results are shown in FIG. More than 50% of CPN-267 remained after 180 minutes. On the other hand, CPN-170, which is a comparative example, had a residual rate of 45% or less after 10 minutes and about 12% after 20 minutes.

(CPN-124, CPN-223)
The stability of CPN-124 (Synthesis Example 34) and CPN-223 (Synthesis Example 16) in human serum was measured over time according to the same method as above. The results are shown in FIG. CPN-223 remained approximately 60% after 180 minutes. On the other hand, CPN-124, which is a comparative example, had a residual rate of about 32% after 20 minutes and about 8% after 40 minutes.

<Rat blood dynamics analysis>
(CPN-170, CPN-267)
In order to analyze the kinetics of CPN-170 (Synthesis Example 36) and CPN-267 (Synthesis Example 22) in rat blood, they were evaluated by the following methods.

A peptide solution (1 mg/100 μL) was administered to Wistar rats (220-250 g) via the jugular vein, and after an arbitrary period of time (1-240 minutes), blood was collected from the previously cannulated femoral artery and subjected to LC/MS. analyzed the blood kinetics of the peptide.

The results are shown in FIG. As can be seen from the results shown in FIG. 7, CPN-267 exhibited better blood kinetics in rat blood than CPN-170. On the other hand, CPN-170 disappeared from rat blood more rapidly than CPN-267.

<Evaluation of weight gain suppression activity in subcutaneous administration to mice>
(CPN-267)
To verify the effect of subcutaneous administration (s.c.) of CPN-267 (Synthesis Example 22) on weight gain in mice, evaluation was performed by the following method.

CPN-267 was subcutaneously administered to ddy male mice (28-31 g) three times (0, 24, 48 hours) at a dose of 6.8 mg/kg (physiological saline was similarly administered as a control). Mice were weighed at 0, 12, 36, 60, 84 and 108 hours after the first administration.

The results are shown in FIG. As can be seen from the results shown in FIG. 8, CPN-267 significantly suppressed body weight gain in mice when administered subcutaneously.

This application is based on Japanese Patent Application No. 2017-8302 filed on January 20, 2017, the disclosure of which is incorporated by reference in its entirety.

Claims (8)

A peptide or a pharmaceutically acceptable salt thereof, represented by the following formula (1) and having a selective agonist activity for type 1 neuromedin U receptor :

In the above formula (1),
X ⁰ is a 3-thienylalanine residue, or is absent;
X ¹ is Trp residue, 3-(benzothiophenyl)alanine residue, 4-benzoyl-phenylalanine residue, 3,3-diphenylalanine residue, 3-naphthyl-alanine residue, 1-methyltryptophan residue , or is absent;
X ² is a Trp residue, a 3-naphthyl-alanine residue, a 4-bromo-phenylalanine residue, a 4-methoxy-phenylalanine residue, a 4-nitro-phenylalanine residue, a 4-trifluoromethyl-phenylalanine residue, 4-phenyl-phenylalanine residue, 4-benzoyl-phenylalanine residue, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid residue, 1,2,3,4-tetrahydronorharman-3-carboxylic acid residue a residue, a 1-methyltryptophan residue, or an α-methyltryptophan residue;
X ³ is an Arg residue, a norarginine residue, or a homoarginine residue;
^X4 is a Pro residue or a homoproline residue;
Z ₁ represents a hydrogen atom, a hydrocarbon group, a heterocyclic group, an acyl group, a sulfo group, a carboxyl group, a glyoxyl group, a polyalkylene glycol group, or a carrier; and Z ₂ represents an amino group, a hydrogen atom, a hydroxy group , an alkoxy group, a hydrocarbon group, a polyalkylene glycol group, or a carrier. X ² is a Trp residue, 3-(2-naphthyl)alanine residue, 4-phenyl-phenylalanine residue, 4-benzoyl-phenylalanine residue, α-methyltryptophan residue, or 1,2,3,4 -tetrahydroisoquinoline-3-carboxylic acid residue and ^X2 is a Trp residue or an α-methyltryptophan residue when ^X1 is an amino acid residue other than a Trp residue. or a pharmaceutically acceptable salt thereof. 3. The peptide or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein ^X1 is a Trp residue, a 3-benzothiophenyl-alanine residue, or a 1-methyltryptophan residue. The peptide according to any one of claims 1 to 3, wherein Z ₁ is a hydrogen atom, an aliphatic acyl group having 1 to 6 carbon atoms, or a group represented by the following formula (3), or a pharmaceutical agent thereof acceptable salts for;

However, in the above formula (3), R ₁ is a substituted or unsubstituted thienyl group, furanyl group, pyrrolidinyl group, pyrrolyl group, imidazolyl group, imidazolinyl group, pyrazolyl group, oxazolyl group, thiazolyl group, cyclopentyl group, cyclo a hexyl group, a phenyl group, a 3-pyridyl group, or a 4-pyridyl group; R ₂ is a single bond, an alkylene group having 1 to 5 carbon atoms, an oxyalkylene group having 1 to 4 carbon atoms, or 1 carbon atom; ∼4 alkyleneoxy groups. The peptide or a pharmaceutically acceptable salt thereof according to claim 1, wherein the peptide represented by formula (1) satisfies the following (i):
(i) X ⁰ -X ¹ -X ² -X ³ -X ⁴ -Arg-Asn in formula (1) above consists of an amino acid sequence selected from the group consisting of SEQ ID NOS: 1-21. The peptide of claim 1, wherein the peptide represented by formula (1) is represented by any one of the following formulas (1-1) to (1-185), or a pharmaceutically acceptable peptide thereof salt:

An activator of type 1 neuromedin U receptor, comprising the peptide according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof as an active ingredient. Metabolic syndrome, obesity, diabetes, dyslipidemia, cardiovascular disease, or circadian rhythm disorder, comprising the peptide according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof as an active ingredient , prophylactic and/or therapeutic agents.

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