WO2015083816A1 - New nk3 receptor agonist - Google Patents

Abstract

　Provided is a new NK3 receptor selective ligand which is expected to have high activity and high selectivity, and to act in a sustained manner in vivo.　A compound expressed by formula (I) X1 - A2 - A3 - X4 - X5 - Met - NH₂ [the symbols in the formula are as set forth in the description] or a pharmaceutically acceptable salt thereof has a highly active and highly selective NK3 receptor agonist activity, and is effective as a drug for the treatment of disorders associated with an excess or deficiency in the secretion of tachykinins, for example as a pharmaceutical for the treatment of pain or for the improvement of sexual dysfunction accompanying excess or deficiency in the secretion of sex hormones.

Description

Novel NK3 receptor agonist

The present invention relates to a peptide having a neurokinin (NK) receptor agonistic action and use thereof. More specifically, the present invention relates to a peptide having a selective and strong binding affinity to NK3 receptor and an agonistic action, and capable of acting continuously in vivo, and uses thereof.

The kisspeptin neuron population in front of the hypothalamus is an ovulation center that regulates the surge-like secretion of gonadotropin-releasing hormone (GnRH) by the positive feedback action of estrogen, and kisspeptins are expected as new drugs that regulate ovulation induction Has been.
On the other hand, various reports on pulsatile secretion of GnRH, which are thought to be involved in follicular development, have recently been made, and kisspeptin neurons in the hypothalamic arcuate nucleus simultaneously express neurokinin B (NKB) and dynorphin ( Non-patent document 1), it has been reported that it plays a role as a center that promotes pulsed secretion of GnRH (non-patent document 2). In addition, the firing activity in this neuron was successfully recorded as a transient rise (MUA volley) of multineuron firing activity (MUA), which is completely synchronized with the luteinizing hormone (LH) pulse. (Non-Patent Documents 3 and 4).
Under these circumstances, the present inventors assume that NK3 receptor ligands (agonists) known as NKB receptors can be agents that regulate GnRH and LH pulses and control follicular development, Started to create a novel NK3 receptor ligand.
Examples of NK3 receptor selective agonists reported so far include [MePhe ⁷ ] -NKB and senktide (Non-patent Documents 5 and 6). These peptides that are already on the market as research reagents have been used for various basic studies involving tachykinins and neurokinin receptors. The present inventors have so far conducted structure-activity relationship studies for the purpose of obtaining knowledge about the structure-requiring properties of [MePhe ⁷ ] -NKB activity and receptor selectivity, and in this process, highly active against NK3 receptor. Moreover, a novel NKB derivative that acts with high selectivity has been found (Patent Document 1, Non-Patent Document 7).

International Publication No. 2014/061772

An object of the present invention is to provide a novel NK3 receptor-selective ligand that is expected to act with high activity, high selectivity, and sustained in vivo.

So far, senktide has been reported as one of NK3 receptor selective agonists. Senktide is a peptidic ligand that exhibits high agonist activity and receptor selectivity for the NK3 receptor, but its discovery was discovered by chance in the creation of an NK1 receptor selective ligand with substance P as a lead compound. Therefore, detailed structure-activity relationship studies of senktide have not been conducted.
The present inventors chemically synthesized a derivative of senktide based on the structure-activity relationship information of tachykinins reported so far, and evaluated the biological activity for three types of human neurokinin receptors in vitro. Moreover, the in vivo biological activity of the synthesized senktide derivative was evaluated by using the response change of goat MUA volley after peripheral administration of the senktide derivative as an index.
As a result, the derivatives of senktide converted from Asp ¹ to L-Glu, D-Glu, and L-Aad, and the N-terminal succinyl group to oxalyl group, etc., are selective only for the NK3 receptor at a lower concentration than senktide. It was found to show binding affinity. All of these derivatives induced an increase in intracellular Ca ²⁺ concentration in the NK3 receptor-expressing cell line, and were confirmed to be potent NK3 receptor agonists. Moreover, it was confirmed by animal experiments using goats that these derivatives show central effects by peripheral administration, similar to senktide.
Subsequently, the present inventors examined the degradation characteristics of senktide against neprilysin known as NKB-degrading enzyme, and senktide is cleaved between Gly-Leu by neprilysin, which is a degrading enzyme of neurokinin B (NKB). Was revealed. Based on this degradation property, we designed and synthesized derivatives in which peptide bonds between senktide and Gly-Leu were converted to various peptide equivalents. As a result, the inventors have found a novel derivative exhibiting high stability against neprilysin while maintaining the same biological activity as senktide, and have completed the present invention.
That is, the present invention is as follows.
[1] Formula (I)

[Wherein X1 represents R1-A1 or a dicarboxylic acid residue;
R1 is a hydrogen atom, acetyl group, oxalyl group, succinyl group, aminocarbonyl group, hydroxyaminocarbonyl group, hydrazinocarbonyl group, methoxydicarbonyl group, aminodicarbonyl group, aminosulfonyl group, N, N-dicarboxymethyl Represents aspartic acid or N-carboxymethylaspartic acid;
A1 represents an L-aspartic acid, D-aspartic acid, L-glutamic acid, D-glutamic acid, or L-α-aminoadipic acid residue;
A2 represents L-phenylalanine, L-tyrosine, L-aspartic acid, D-aspartic acid, L-glutamic acid, or D-glutamic acid residue;
A3 represents LN-methylphenylalanine, LN-methylvaline, LN-methylisoleucine, LN-methyltyrosine, or LN-methyltryptophan residue;
X4-X5 represents Gly-Leu or its dipeptide equivalent;
Gly represents a glycine residue;
Leu represents a leucine residue;
Met-NH ₂ represents methionine amide] or a physiologically acceptable salt thereof.
[2] X1 is

Wherein R ^a and R ^{a ′} are the same or different and are a succinyl group or an oxalyl group; R ^b and R ^{b ′} are the same or different and are a succinyl group or an oxalyl group; R ^c and R ^{c ′} are the same Or a succinyl group or an oxalyl group; and R ^d and R ^{d ′} are the same or different and are a succinyl group or an oxalyl group).
The compound of the above-mentioned [1] or a physiologically acceptable salt thereof.
[3] The dipeptide equivalent is

The compound according to [1] or [2] above, or a physiologically acceptable salt thereof.
[4] The compound according to the above [1], or a physiologically acceptable salt thereof, wherein the compound or the physiologically acceptable salt thereof is any one peptide or physiologically acceptable salt represented by the following sequence: Acceptable salts;

[5] The compound according to the above [1] or physiologically acceptable salt thereof, wherein the compound or physiologically acceptable salt thereof is any one peptide or physiologically acceptable salt represented by the following sequence: Acceptable salts;

[6] A medicament comprising the compound according to any one of [1] to [5] above or a physiologically acceptable salt thereof.
[7] The medicament of the above-mentioned [6], which is a neurokinin receptor agonist.
[8] The medicament according to [7] above, wherein the neurokinin receptor is an NK3 receptor.
[9] The medicament according to [6] above, wherein tachykinins and / or neurokinin receptors are therapeutic agents for diseases associated with the onset and progression thereof.
[10] The medicament according to [9] above, wherein the disease is sexual dysfunction associated with pain or excessive / deficiency of sex hormone secretion.
[11] Formula (I) for treating diseases in which tachykinins and / or neurokinin receptors are involved in the onset and progression thereof

[Wherein X1 represents R1-A1 or a dicarboxylic acid residue;
R1 is a hydrogen atom, acetyl group, oxalyl group, succinyl group, aminocarbonyl group, hydroxyaminocarbonyl group, hydrazinocarbonyl group, methoxydicarbonyl group, aminodicarbonyl group, aminosulfonyl group, N, N-dicarboxymethyl Represents aspartic acid or N-carboxymethylaspartic acid;
A1 represents an L-aspartic acid, D-aspartic acid, L-glutamic acid, D-glutamic acid, or L-α-aminoadipic acid residue;
A2 represents L-phenylalanine, L-tyrosine, L-aspartic acid, D-aspartic acid, L-glutamic acid, or D-glutamic acid residue;
A3 represents LN-methylphenylalanine, LN-methylvaline, LN-methylisoleucine, LN-methyltyrosine, or LN-methyltryptophan residue;
X4-X5 represents Gly-Leu or its dipeptide equivalent;
Gly represents a glycine residue;
Leu represents a leucine residue;
Met-NH ₂ represents methionine amide] or a physiologically acceptable salt thereof.
[12] The compound of the above-mentioned [11] or a physiologically acceptable salt thereof, wherein the disease is sexual dysfunction associated with pain or excessive / deficiency of sex hormone secretion.
[13] A tachykinin comprising administering an effective amount of the compound according to any one of [1] to [5] above or a physiologically acceptable salt thereof to a subject in need thereof, and A method for treating a disease in which a neurokinin receptor is involved in its onset or progression.
[14] The method described in [13] above, wherein the disease is sexual dysfunction associated with pain or excess / deficiency of sex hormone secretion.
[15] A reagent comprising the compound according to any one of [1] to [5] above or a physiologically acceptable salt thereof.
[16] The reagent according to [15] above, wherein tachykinins and / or neurokinin receptors are used for diagnosis and / or research of diseases related to the onset and progression thereof.

The compound of the present invention has high stability against neprilysin while maintaining a biological activity equivalent to or higher than that of senktide. Therefore, the highly active and highly selective NK3 receptor agonist obtained by the present invention is a therapeutic agent for diseases associated with excessive or deficient secretion of tachykinins, such as pain, sex function associated with excessive or deficient sex hormone secretion. It is useful as a drug for improving disorders, a reagent for basic science experiments for developing therapeutics for these diseases, and the like.

It is a figure which shows the result of having compared the stability with respect to neprilysin of Senktide and the Senktide derivative (E14) of this invention. a) Schematic diagram showing the cleavage site of Senktide by neprilysin. b) It is the figure which investigated decomposition | disassembly of Senktide by Neprilysin and the Senktide derivative (E14) of this invention with time. Senktide (●), N-terminal fragment (▲), C-terminal fragment (Δ), E14 (◯). mean ± SD (n = 3)

Hereinafter, the definition of each symbol used in this specification will be described in detail.
In the present specification, amino acids and the like are represented by abbreviations based on abbreviations by IUPAC-IUB Commission on Biochemical Nomenclature or conventional abbreviations in the field, examples of which are described below. In addition, when there are optical isomers for amino acids, “L-” indicates L-form (for example, “L-Ala” is L-form Ala), and “D-” indicates “D-”. (For example, “D-Ala” is D-form Ala), and “DL-” is D-form and L-form racemate (eg “DL-Ala” is D-form) A racemic mixture of Ala and L Ala). In the present invention, the amino acid residue may be D-form or L-form, but is preferably L-form.
Gly or G: Glycine Ala or A: Alanine Val or V: Valine Leu or L: Leucine Ile or I: Isoleucine Ser or S: Serine Thr or T: Threonine Cys or C: Cysteine Met or M: Methionine Glu or E: Glutamic acid Asp or D: Aspartic acid Lys or K: Lysine Arg or R: Arginine His or H: Histidine Phe or F: Phenylalanine Tyr or Y: Tyrosine Trp or W: Tryptophan Pro or P: Proline Asn or N: Asparagine Gln or Q: Glutamine pGlu: pyroglutamic acid MePhe: N-methylphenylalanine MeVal: N-methylvaline MeIle: N-methylisoleucine MeTyr : N-methyltyrosine MeTrp: N-methyltryptophan MeLeu: N-methylleucine MeAsp: N-methylaspartic acid Aad: 2-aminoadipic acid Succinyl: -CO-CH ₂ CH ₂ -COOH
Oxalyl: -CO-COOH
Ac: Acetyl group (-COCH ₃ )

The present invention provides a compound having an agonistic action specific to a neurokinin receptor, specifically, an NK3 receptor. Specifically, the formula (I)

[Wherein X1 represents R1-A1 or a dicarboxylic acid residue;
R1 is a hydrogen atom, acetyl group, oxalyl group, succinyl group, aminocarbonyl group, hydroxyaminocarbonyl group, hydrazinocarbonyl group, methoxydicarbonyl group, aminodicarbonyl group, aminosulfonyl group, N, N-dicarboxymethyl Represents aspartic acid or N-carboxymethylaspartic acid;
A1 represents an L-aspartic acid, D-aspartic acid, L-glutamic acid, D-glutamic acid, or L-α-aminoadipic acid residue;
A2 represents L-phenylalanine, L-tyrosine, L-aspartic acid, D-aspartic acid, L-glutamic acid, or D-glutamic acid residue;
A3 represents LN-methylphenylalanine, LN-methylvaline, LN-methylisoleucine, LN-methyltyrosine, or LN-methyltryptophan residue;
X4-X5 represents Gly-Leu or its dipeptide equivalent;
Gly represents a glycine residue;
Leu represents a leucine residue;
Met-NH ₂ represents methionine amide] (also referred to herein as compound (I)) or a physiologically acceptable salt thereof.

In the above formula (I), X1 represents R1-A1 or a dicarboxylic acid residue. Here, R1 is a hydrogen atom, acetyl group, oxalyl group, succinyl group, aminocarbonyl group, hydroxyaminocarbonyl group, hydrazinocarbonyl group, methoxydicarbonyl group, aminodicarbonyl group, aminosulfonyl group, N, N— Dicarboxymethyl aspartic acid or N-carboxymethyl aspartic acid, and A1 represents an L-aspartic acid, D-aspartic acid, L-glutamic acid, D-glutamic acid, or L-α-aminoadipic acid residue. To express. The dicarboxylic acid residue as X1 is not particularly limited as long as it has a structure having two carboxyl groups. Preferable X1 includes a group represented by the following structure.

In the above formula, R ^a and R ^{a ′} are the same or different and are a succinyl group or an oxalyl group, and preferably, R ^a and R ^{a ′} are the same and are a succinyl group or an oxalyl group. R ^b and R ^{b ′} are the same or different and are a succinyl group or an oxalyl group. Preferably, R ^b and R ^{b ′} are the same and are a succinyl group or an oxalyl group. R ^c and R ^{c ′} are the same or different and are a succinyl group or an oxalyl group. Preferably, R ^c and R ^{c ′} are the same and are a succinyl group or an oxalyl group. R ^d and R ^{d ′} are the same or different and are a succinyl group or an oxalyl group, and preferably R ^d and R ^{d ′} are the same and are a succinyl group or an oxalyl group.

In the above formula (I), A2 represents L-phenylalanine, L-tyrosine, L-aspartic acid, D-aspartic acid, L-glutamic acid, or D-glutamic acid residue.

In the above formula (I), A3 represents LN-methylphenylalanine, LN-methylvaline, LN-methylisoleucine, LN-methyltyrosine, or LN-methyltryptophan residue.

In the above formula (I), X4-X5 represents Gly-Leu or a dipeptide equivalent thereof. Gly-Leu represents a dipeptide residue in which glycine and leucine are bound by a peptide bond, and is represented by the following structure.

The dipeptide equivalent of Gly-Leu is obtained by replacing the peptide bond (-CONH-) of Gly-Leu with another bond, and the other bond includes -CH = CH-, -CH _2- CH _2- , -CH ₂ -CH =, -C (= N-OH) -NH-, -C (= NH) -NH- and the like. Such a dipeptide equivalent is sometimes referred to as GlyψLeu in this specification.

Compound (I) is preferably exemplified by the peptides listed in Tables 1 to 7 below.

40b1 and 40b2, 40d1 and 40d2, and 40g1 and 40g2 are compounds having different stereochemical structures synthesized using a mixture of cis and trans as raw materials, respectively. 112 is a compound obtained using a cis isomer as a raw material, and 113 is a compound synthesized using a mixture of cis and trans.

117 is a compound obtained using a cis isomer as a raw material, and 118 is a compound synthesized using a mixture of cis and trans.

More preferably,

Any one of the peptides represented by the sequence or a physiologically acceptable salt, more preferably

Any one of the peptides represented by the sequence or a physiologically acceptable salt.

Compound (I) may be in the form of a salt. Examples of such salts include metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. Preferable examples of the metal salt include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt, magnesium salt and barium salt; aluminum salt and the like. Preferable examples of the salt with an organic base include, for example, trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N, N′-dibenzylethylenediamine. And the like. Preferable examples of the salt with inorganic acid include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid and the like. Preferable examples of the salt with organic acid include formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, and benzenesulfone. And salts with acid, p-toluenesulfonic acid and the like. Preferable examples of salts with basic amino acids include salts with arginine, lysine, ornithine and the like, and preferable examples of salts with acidic amino acids include salts with aspartic acid and glutamic acid, for example. It is done.
Of these, pharmaceutically acceptable salts are preferred. For example, when the compound has an acidic functional group, an inorganic salt such as an alkali metal salt (eg, sodium salt, potassium salt), an alkaline earth metal salt (eg, calcium salt, magnesium salt, barium salt), When the ammonium salt has a basic functional group in the compound, for example, a salt with an inorganic acid accompanied by hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, or acetic acid, phthalic acid, fumaric acid Further, salts with organic acids such as oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid and p-toluenesulfonic acid are preferred.

Compound (I) is a peptide and can be produced according to a peptide synthesis method known per se. As a peptide synthesis method, for example, either a solid phase synthesis method or a liquid phase synthesis method may be used. That is, the target peptide can be produced by repeatedly condensing the partial peptide or amino acid capable of constituting the compound (I) and the remaining portion in accordance with the desired sequence. When the product having the desired sequence has a protecting group, the target peptide can be produced by removing the protecting group. Examples of known condensation methods and protecting group elimination methods include the methods described in the following (1) to (5).
(1) M. Bodanszky and MA Ondetti, Peptide Synthesis, Interscience Publishers, New York (1966)
(2) Schroeder and Luebke, The Peptide, Academic Press, New York (1965)
(3) Nobuo Izumiya et al., Basics and Experiments of Peptide Synthesis, Maruzen Co., Ltd. (1975)
(4) Haruaki Yajima and Shunpei Sugawara, Biochemistry Experiment Course 1, Protein Chemistry IV, 205, (1977)
(5) Supervised by Yajima Haruaki, Development of follow-up medicines Volume 14: Peptide synthesis Hirokawa Shoten Also, after the reaction, a combination of conventional purification methods such as solvent extraction, distillation, column chromatography, liquid chromatography, recrystallization, etc. The peptide of the present invention can be purified and isolated. When the peptide obtained by the above method is a free form, it can be converted into an appropriate salt by a known method. Conversely, when it is obtained as a salt, it can be converted into a free form by a known method. .
The raw material compound may be a salt, and examples of such a salt include the same as those described above as the salt of the peptide of the present invention.

Regarding the condensation of protected amino acids or peptides, various activating reagents that can be used for peptide synthesis can be used, and triphosphonium salts, tetramethyluronium salts, carbodiimides, and the like are particularly preferable. Trisphosphonium salts include benzotriazol-1-yloxytris (pyrrolidino) phosphonium hexafluorophosphate (PyBOP), bromotris (pyrrolidino) phosphonium hexafluorophosphate (PyBroP), 7-azabenzotriazole- 1-yloxytris (pyrrolidino) phosphonium hexafluorophosphate (PyAOP), tetramethyluronium salts are 2- (1H-benzotriazol-1-yl) -1,1,3,3-hexafluoro Phosphate (HBTU), 2- (7-azabenzotriazol-1-yl) -1,1,3,3-hexafluorophosphate (HATU), 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2- (5-norbornene-2,3-dicarboximide) -1,1,3,3-tetramethyluronium tetra Fluoroborate (TNTU), O -(N-succimidyl) -1,1,3,3-tetramethyluronium tetrafluoroborate (TSTU), carbodiimides include DCC, N, N'-diisopropylcarbodiimide (DIPCDI), N-ethyl-N ' -(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI · HCl) and the like. These condensations include racemization inhibitors such as N-hydroxy-5-norbornene-2,3-dicarboximide (HONB), 1-hydroxybenzotriazole (HOBt), 1-hydroxy-7-azabenzotriazole ( HOAt), 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine (HOOBt) etc.) is preferred. The solvent used for the condensation may be appropriately selected from solvents that are known to be usable for peptide condensation reactions. For example, anhydrous or hydrous acid amides such as N, N-dimethylformamide, N, N-dimethylacetamide and N-methylpyrrolidone, halogenated hydrocarbons such as methylene chloride and chloroform, alcohols such as trifluoroethanol and phenol , Sulfoxides such as dimethyl sulfoxide, tertiary amines such as pyridine, ethers such as dioxane and tetrahydrofuran, nitriles such as acetonitrile and propionitrile, esters such as methyl acetate and ethyl acetate, or appropriate mixtures thereof, etc. Is used. The reaction temperature is appropriately selected from a range known to be usable for peptide bond formation reaction, and is usually selected appropriately from a range of about −20 ° C. to 50 ° C. The activated amino acid derivative is usually used in an excess of 1.5 to 6 times. In the case of solid phase synthesis, as a result of a test using the ninhydrin reaction, if the condensation is insufficient, sufficient condensation can be performed by repeating the condensation reaction without removing the protecting group. If sufficient condensation cannot be obtained even after repeating the reaction, the unreacted amino acid can be acylated with acetic anhydride or acetylimidazole so as not to affect the subsequent reaction.

Examples of the protecting group for the amino group of the starting amino acid include benzyloxycarbonyl (Z), tert-butoxycarbonyl (Boc), tert-pentyloxycarbonyl, isobornyloxycarbonyl, 4-methoxybenzyloxycarbonyl, Cl-Z , Br-Z, adamantyloxycarbonyl, trifluoroacetyl, phthaloyl, formyl, 2-nitrophenylsulfenyl, diphenylphosphinothioyl, 9-fluorenylmethyloxycarbonyl (Fmoc), trityl (Trt), etc. .
Examples of the protecting group for the carboxyl group of the raw material amino acid include a tert-butyl (Bu ^t ) group, a benzyl (Bzl) group, a C _1-6 alkyl group, a C _3-10 cycloalkyl group, and a C _7-14 aralkyl group. Other examples include allyl, 2-adamantyl, 4-nitrobenzyl, 4-methoxybenzyl, 4-chlorobenzyl, phenacyl and benzyloxycarbonyl hydrazide, tert-butoxycarbonyl hydrazide, trityl hydrazide and the like.
The hydroxyl groups of serine and threonine can be protected, for example, by esterification or etherification. Examples of the group suitable for esterification include a lower (C _2-4 ) alkanoyl group such as an acetyl group, an aroyl group such as a benzoyl group, and a group derived from an organic acid. Examples of groups suitable for etherification include benzyl, tetrahydropyranyl, Bu ^t , Trt and the like.
Examples of groups for protecting the phenolic hydroxyl group of tyrosine include Bzl, 2,6-di-chlorobenzyl, 2-nitrobenzyl, Br-Z, Bu ^t, and the like.
Examples of the protecting group for imidazole of histidine include p-toluenesulfonyl (Tos), 4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr), 2,4-dinitrophenyl (DNP), benzyloxymethyl (Bom ), Tert-butoxymethyl (Bum), Boc, Trt, Fmoc and the like.
Examples of protecting groups for the guanidino group of arginine include Tos, Z, 4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr), p-methoxybenzenesulfonyl (MBS), 2,2,5,7,8. -Pentamethylchroman-6-sulfonyl (Pmc), mesitylene-2-sulfonyl (Mts), 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (Pbf), Boc, Z, NO ₂ etc. Is mentioned.
Examples of the protecting group for the side chain amino group of lysine include Z, Cl-Z, trifluoroacetyl, Boc, Fmoc, Trt, Mtr, 4,4-dimethyl-2,6-dioxocyclohexylideneyl (Dde ) And the like.
Examples of indolyl protecting groups for tryptophan include formyl (For), Z, Boc, Mts, Mtr and the like.
Examples of protecting groups for asparagine and glutamine include Trt, xanthyl (Xan), 4,4′-dimethoxybenzhydryl (Mbh), 2,4,6-trimethoxybenzyl (Tmob) and the like.
Examples of the activated carboxyl group of the raw material include the corresponding acid anhydride, azide, active ester [alcohol (eg, pentachlorophenol, 2,4,5-trichlorophenol, 2,4-dinitrophenol, cyano Methyl alcohol, paranitrophenol, HONB, N-hydroxysuccinimide, HOBt, ester with HOAt) and the like. Examples of the activated amino group of the raw material include a corresponding phosphite amide.

Examples of methods for removing (eliminating) protecting groups include catalytic reduction in a hydrogen stream in the presence of a catalyst such as Pd black or Pd carbon, and anhydrous hydrogen fluoride, methanesulfonic acid, trifluoromethanesulfonic acid. , Acid treatment with trifluoroacetic acid, trimesylsilane bromide (TMSBr), trimethylsilyl trifluoromethanesulfonate, tetrafluoroboric acid, tris (trifluoro) boron, boron tribromide or a mixture thereof, diisopropylethylamine, triethylamine , Base treatment with piperidine, piperazine, etc., reduction with sodium in liquid ammonia, and the like. The elimination reaction by the acid treatment is generally carried out at a temperature of −20 ° C. to 40 ° C. In the acid treatment, a cation scavenger such as anisole, phenol, thioanisole, metacresol, paracresol, dimethyl sulfide, 1 Addition of 1,4-butanedithiol, 1,2-ethanedithiol, etc. is effective. The 2,4-dinitrophenyl group used as the imidazole protecting group of histidine was removed by thiophenol treatment, and the formyl group used as the indole protecting group of tryptophan was the above 1,2-ethanedithiol and 1,4-butane. In addition to deprotection by acid treatment in the presence of dithiol or the like, it can also be removed by alkali treatment with dilute sodium hydroxide, dilute ammonia or the like.
The protection of the functional group that should not be involved in the reaction of the raw material and the protection group, the elimination of the protective group, the activation of the functional group involved in the reaction, etc. can be appropriately selected from known protective groups or known means.

Compound (I) has methionine amide at the C-terminus. Peptide amides can be obtained by solid-phase synthesis using a resin for amide synthesis or by amidating the α-carboxyl group of the carboxyl-terminal amino acid, and then connecting the peptide chain to the desired chain length on the amino group side. After the extension, a peptide in which only the N-terminal α-amino group protecting group of the peptide chain is removed and a peptide (or amino acid) in which only the C-terminal carboxyl group protecting group is removed are prepared. Condensation in a mixed solvent as described above. The details of the condensation reaction are the same as described above. After purifying the protected peptide obtained by the condensation, all the protecting groups are removed by the above-described method to obtain the desired crude polypeptide. This crude peptide can be purified using various known purification means, and the main fraction can be lyophilized to obtain an amide of the desired peptide.

When compound (I) exists as a configurational isomer (configuration isomer) such as an enantiomer or diastereomer, a conformer (conformation isomer), etc., these are also contained as compound (I), If desired, each can be isolated by means known per se, the above-described separation and purification means. In addition, when the compound (I) is a racemate, it can be separated into an S form and an R form by an ordinary optical resolution means.
When a stereoisomer exists in the compound (I), the case where this isomer is used alone or a mixture thereof is also included in the compound (I).
Further, the compound (I) may be a solvate (eg, hydrate) or a non-solvate (eg, non-hydrate).
Compound (I) may be labeled with an isotope (eg, ³ H, ¹⁴ C, ³⁵ S, ¹²⁵ I) or the like.
Further, the compound (I) may be a deuterium converter obtained by converting ¹ H into ² H (D).

The peptide in the present specification has an N-terminus (amino terminus) at the left end and a C-terminus (carboxyl terminus) at the right end according to the convention of peptide designation. When “H—” is marked in the N-terminal amino acid of the peptide, it indicates that the terminal amino group is not derivatized. When “—NH ₂ ” is marked in the C-terminal amino acid of the peptide, it indicates that the terminal carboxyl group is amidated.

Compound (I) may be a crystal, and the crystal form of the crystal may be single or plural. Crystals can be produced using a crystallization method known per se.
Compound (I) may be a pharmaceutically acceptable cocrystal or cocrystal salt. Here, co-crystals or co-crystal salts are two or more unique at room temperature, each having different physical properties (eg structure, melting point, heat of fusion, hygroscopicity, solubility and stability). It means a crystalline substance composed of a simple solid. The cocrystal or cocrystal salt can be produced according to a cocrystallization method known per se.
The crystals of compound (I) are excellent in physicochemical properties (eg, melting point, solubility, stability) and biological properties (eg, pharmacokinetics (absorbability, distribution, metabolism, excretion), drug efficacy), pharmaceuticals As extremely useful.

Compound (I) has an NK3 receptor agonistic action. From this action, compound (I) can react with mammals (eg, humans, monkeys, cats, pigs, horses, cows, mice, rats, guinea pigs, dogs, rabbits, etc.) tachykinins and / or neurokinin receptors. Is useful as a preventive or therapeutic agent for diseases and pathological conditions related to its onset and progression. Accordingly, the present invention provides an effective amount of Compound (I) to a subject in need thereof (eg, human, monkey, cat, pig, horse, cow, mouse, rat, guinea pig, dog, rabbit, etc .; particularly human) A method for treating a disease in which tachykinins and / or neurokinin receptors are involved in the onset and progression thereof.
Examples of such diseases and pathologies include sexual dysfunction / developmental failure due to excessive or insufficient sex hormone secretion, benign tumors, malignant tumors, metabolic abnormalities, and the like.

Compound (I) can be used as a medicine by formulation according to a method known per se, for example, a method described in the Japanese Pharmacopoeia, as it is or with a pharmacologically acceptable carrier.

The medicament comprising compound (I) has low toxicity, and according to means known per se generally used in the production of pharmaceutical preparations, compound (I) can be used as it is or as a pharmacologically acceptable carrier. For example, tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets), powders, granules, capsules (including soft capsules and microcapsules), liquids, troches, and syrups , Emulsion, suspension, injection (eg, subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection, etc.), external preparation (eg, nasal preparation, transdermal preparation, ointment) , Suppositories (eg, rectal suppositories, vaginal suppositories), pellets, nasal preparations, pulmonary preparations (inhalants), drops, and the like, orally or parenterally (eg, topical, rectal, Intravenous administration can be safely administered.
These preparations may be controlled-release preparations (eg, sustained-release microcapsules) such as immediate-release preparations or sustained-release preparations.
The content of compound (I) in the pharmaceutical preparation is about 0.01 to about 100% by weight of the whole preparation.
The dose of compound (I) is appropriately selected depending on the administration subject, symptoms, administration method and the like. For example, when the compound of the present invention is administered orally, the dose for humans (with a body weight of 60 kg) is about 0.1 to 100 mg, preferably about 1.0 to 50 mg, more preferably about 1.0 to 20 mg per day. It is. When Compound (I) is administered parenterally, the dose for humans (with a body weight of 60 kg) is about 0.01 to 30 mg, preferably about 0.1 to 20 mg, more preferably about 0.5, per day. ~ 10 mg. This amount can be administered one to several times a day.

Examples of the pharmacologically acceptable carrier that may be used in the production of the medicament of the present invention include various organic or inorganic carrier substances that are conventionally used as a preparation material. For example, excipients, lubricants in solid preparations, Examples include binders and disintegrants, solvents in liquid preparations, solubilizers, suspending agents, tonicity agents, buffers, and soothing agents. If necessary, additives such as conventional preservatives, antioxidants, colorants, sweeteners, adsorbents, wetting agents and the like can be used in appropriate amounts.
Examples of the excipient include lactose, sucrose, D-mannitol, starch, corn starch, crystalline cellulose, light anhydrous silicic acid and the like.
Examples of the lubricant include magnesium stearate, calcium stearate, talc, colloidal silica and the like.
Examples of the binder include crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, starch, sucrose, gelatin, methylcellulose, sodium carboxymethylcellulose and the like.
Examples of the disintegrant include starch, carboxymethyl cellulose, carboxymethyl cellulose calcium, carboxymethyl starch sodium, L-hydroxypropyl cellulose, and the like.
Examples of the solvent include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil, olive oil and the like.
Examples of the solubilizer include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like.
Examples of the suspending agent include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glyceryl monostearate; for example, polyvinyl alcohol, polyvinylpyrrolidone, carboxy Examples include hydrophilic polymers such as sodium methylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose.
Examples of the isotonic agent include glucose, D-sorbitol, sodium chloride, glycerin, D-mannitol and the like.
Examples of the buffer include buffer solutions of phosphate, acetate, carbonate, citrate and the like.
Examples of soothing agents include benzyl alcohol.
Examples of the preservative include parahydroxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like.
Examples of the antioxidant include sulfite, ascorbic acid, α-tocopherol and the like.
Examples of the colorant include water-soluble edible tar dyes (eg, edible dyes such as edible red Nos. 2 and 3, edible yellows Nos. 4 and 5, edible blue Nos. 1 and 2), water-insoluble lake dyes (eg, Examples thereof include aluminum salts of water-soluble edible tar pigments, natural pigments (eg, β-carotene, chlorophyll, bengara) and the like.
Examples of the sweetening agent include saccharin sodium, dipotassium glycyrrhizinate, aspartame, stevia and the like.
Examples of the adsorbent include porous starch, calcium silicate (trade name: Florite RE), magnesium aluminate metasilicate (trade name: Neusilin), and light anhydrous silicic acid (trade name: Cylicia).
Examples of the wetting agent include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylene lauryl ether.

When manufacturing an oral preparation, it may be coated for the purpose of taste masking, enteric solubility or sustainability, if necessary.

Examples of the coating base used for coating include sugar coating base, water-soluble film coating base, enteric film coating base and sustained-release film coating base.

As the sugar coating base, sucrose is used, and one or more selected from talc, precipitated calcium carbonate, gelatin, gum arabic, pullulan, carnauba wax and the like may be used in combination.

Examples of the water-soluble film coating base include cellulose polymers such as hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, and methylhydroxyethylcellulose; polyvinyl acetal diethylaminoacetate, aminoalkyl methacrylate copolymer E [Eudragit E (trade name) ], Synthetic polymers such as polyvinylpyrrolidone; polysaccharides such as pullulan.

Examples of enteric film coating bases include cellulose polymers such as hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, carboxymethylethylcellulose, and cellulose acetate phthalate; methacrylic acid copolymer L [Eudragit L (trade name) ] Acrylic acid polymers such as methacrylic acid copolymer LD [Eudragit L-30D55 (trade name)], methacrylic acid copolymer S [Eudragit S (trade name)]; natural products such as shellac.

Examples of the sustained-release film coating base include cellulose polymers such as ethyl cellulose; aminoalkyl methacrylate copolymer RS [Eudragit RS (trade name)], ethyl acrylate-methyl methacrylate copolymer suspension [Eudragit Acrylic polymer such as NE (trade name)].

The above-mentioned coating bases may be used by mixing two or more of them in an appropriate ratio. Moreover, you may use light-shielding agents, such as a titanium oxide, ferric oxide, etc. in the case of coating.

NK3 receptor is known to be involved in various in vivo phenomena. Therefore, the compound (I) having an NK3 receptor agonistic action can be a useful tool for elucidating various in vivo phenomena involving the NK3 receptor. Therefore, the compound of the present invention is also useful as a research reagent.

The present invention is further illustrated in detail by the following examples and test examples, which are merely implementations and are not intended to limit the present invention, and may be varied without departing from the scope of the present invention. May be.

Example 1 Construction of Protected Peptide Resin On NovaSyn® TGR resin (100 mg, 0.026 mmol) or Rink Amide resin (45.5 mg, 0.025 mmol), Fmoc protected amino acid or Fmoc protected dipeptide equivalent (0.075 mmol), HOBt A protected peptide resin was constructed by the Fmoc solid phase synthesis method using H ₂ O (11.5 mg, 0.075 mmol) and DIC (11.6 μL, 0.075 mmol). For acylation to N-methylamino acid, Fmoc protected amino acid (0.125 mmol), HATU (46.8 mg, 0.12 mmol) / (i-Pr) ₂ NEt (43.6 μL, 0.25 mmol) was used.

Example 2 Acetylation of peptide chain N terminal After constructing a protected peptide resin by the procedure of Example 1, acetic anhydride (20.2 μL, 0.25 mmol) and pyridine (20.2 μL, 0.25 mmol) were added in DMF solvent, The target modified protected peptide chain was obtained by reacting for 1.5 hours.

Example 3 Succinylation of peptide chain N-terminus After constructing a protected peptide resin by the procedure of Example 1, succinic anhydride (12.5 mg, 0.125 mmol) and (i-Pr) ₂ NEt (43.6 μL, 0.125 in DMF solvent) mmol) was added and reacted at room temperature for 1.5 hours to obtain the desired modified protected peptide chain.

Example 4 Oxalation of peptide chain N-terminus After constructing a protected peptide resin by the procedure of Example 1, tert-butyl chloro (oxo) acetate (20.6 mg, 0.125 mmol) and (i-Pr) in CH ₂ Cl ₂ solvent ) ₂ NEt (43.6 μL, 0.25 mmol) was added and reacted at room temperature for 1.5 hours to obtain the desired modified protected peptide chain.

Example 5 Carbamoylation of peptide chain N-terminus After construction of a protected peptide resin by the procedure of Example 1, chlorosulfonyl isocyanate (10.9 μL, 0.125 mmol) and pyridine (20.2 μL, 0.25 mmol) were added in CH ₂ Cl ₂ solvent. And allowed to react at room temperature for 16 hours. Thereafter, H ₂ O (50 μL) was added and reacted for 48 hours to obtain the target modified protected peptide chain.

Example 6 Hydroxycarbamoylation of N-terminal of peptide chain After constructing a protected peptide resin by the procedure of Example 1, p-nitrophenyl chloroformate (186.4 mg, 0.925 mmol) in THF / CH ₂ Cl ₂ (1: 1) solvent (i-Pr) ₂ NEt (161.4 μL, 0.925 mmol) was added and reacted at room temperature for 30 minutes. Thereafter, hydroxylamine hydrochloride (64.3 mg, 0.925 mmol) and (i-Pr) ₂ NEt (322.3 μL, 1.85 mmol) were added in DMF solvent and reacted for 15 minutes to obtain the target modified protected peptide chain. It was.

Example 7 Hydrazine Carbonylation of Peptide Chain N-Terminal After constructing a protected peptide resin by the procedure of Example 1, p-nitrophenyl chloroformate (100.8 mg, 0.5 mmol), tert-butyl carbazate (66.1 mg, 0.5 mmol) in anhydrous THF ) And N-methylmorpholine (55 μL, 0.5 mmol) and triethylamine (69.3 μL, 0.5 mmol) are added and reacted at room temperature for 42 hours to obtain the desired modified protected peptide chain (Boc protector). Obtained.

Example 8 Methyloxalylation of N-terminal of peptide chain After construction of protected peptide resin by the procedure of Example 1, methyl oxalyl chloride (11.4 μL, 0.125 mmol) and pyridine (20.1 μL, 0.25 mmol) in CH ₂ Cl ₂ solvent Was added and reacted at room temperature for 2 hours to obtain the desired modified protected peptide chain.

Example 9 Oxidation of peptide chain N-terminal After construction of a protected peptide resin by the procedure of Example 1, oxamic acid (11.1 mg, 0,125 mmol), DIC (19.4 μL, 0.125 mmol) and HOBt in CH ₂ Cl ₂ solvent _{· H 2 O (19.1 mg,} 0.125 mmol) was added, by reacting at room temperature for 2 hours to obtain a modified protected peptide chain of interest.

Example 10 Sulfonamidation of peptide chain N-terminal After constructing a protected peptide resin by the procedure of Example 1, chlorosulfonyl isocyanate (21.8 μL, 0.25 mmol) and tert-butanol (23.7 μL, 0.25 mmol) in anhydrous CH ₂ Cl ₂ solvent. mmol) and triethylamine (69.3 μL, 0.5 mmol) were added and reacted at room temperature for 3 hours to obtain the target modified protected peptide chain (Boc protector).

Example 11 Carboxymethylation of N-terminal of peptide chain After constructing a protected peptide resin by the procedure of Example 1, tert-butyl bromoacetate (18.3 μL, 0.125 mmol) and (i-Pr) ₂ NEt (43.6) in anhydrous DMF solvent μL, 0.25 mmol) was added and reacted at room temperature for 16 hours to obtain the target modified protected peptide chain (tert-Bu protector).

Example 12 N-terminal bis (carboxymethyl) peptide chain N-Fmoc-L-aspartate 4-tertbutyl (2.0 g, 4.86 mmol), triethylamine (1.0 mL, 7.29 mmol) and benzyl bromide in DMF solvent (865.8 μL, 7.29 mmol) were mixed and reacted at room temperature for 14 hours to obtain 4-tertbutyl 1-benzyl N-Fmoc-L-aspartate (1.74 g). By adding diethylamine (2.0 mL, 20.2 mmol) in DMF to the product (700 mg, 1.41 mmol) and reacting at room temperature for 30 minutes, L-aspartate 4-tert-butyl 1-benzyl ( 368.3 mg) was obtained. Then, tert-butyl bromoacetate (919.6 μL, 6.27 mmol) and potassium carbonate (1049.0 mg, 7.59 mmol) were added to the product (368.3 mg, 1.32 mmol) in anhydrous DMF solvent and reacted at room temperature for 40 hours. To give 4-tert-butyl 1-benzyl N, N-bis (tert-butyloxycarbonylmethyl) -L-aspartate (528.8 mg). By adding 10% Pd / C (14.8 mg, 50 mg / mmol) in EtOH solvent to the product (150 mg, 0.30 mmol) and reacting in a hydrogen atmosphere at room temperature for 6.5 hours, N, N-bis (Tert-Butyloxycarbonylmethyl) -L-aspartate 4-tert-butyl (113.5 mg) was obtained. This was applied to the Fmoc solid phase synthesis method of Example 1 and introduced into a protected peptide resin to construct the desired modified protected peptide resin (tert-Bu protector).

Example 13 Deprotection and Purification of Protected Peptide Resin The obtained resin was treated with ice in a reaction solution of TFA / m-cresol / thioanisole / 1,2-ethanedithiol / H ₂ O (80: 5: 5: 5: 5). The mixture was treated for 2 hours under cooling, and cut out from the resin accompanied by removal of side chain protecting groups. The resin was removed by filtration, and TFA in the filtrate was distilled off by azeotropic distillation with toluene, and then cold Et ₂ O was added to precipitate the peptide. The obtained crude peptide was washed three times and then purified by HPLC to obtain the target peptide. The physical property data of the obtained peptide is shown in Table 8 below. The structure of each peptide corresponds to the structure of each peptide number in Tables 1-7. Hereinafter, the compound (peptide) obtained by the present invention is also referred to as a senktide derivative of the present invention.

Senktide is a known NK3 receptor agonist.

Test Example 1 Evaluation of NK3 receptor binding inhibitory activity of radioactive NKB by Senktide derivative of the present invention Flp-In CHO cells forcibly expressing NK3 receptor (In order to stably express NK3 receptor in Flp-In CHO cells of Invitrogen) the receptor membrane fraction prepared from transgenic to those prepared) were used to evaluate the various peptides ^{([125 I] His 3,} MePhe 7) -NKB receptor binding inhibitory activity.
First, 50 μL of a compound (peptide) solution prepared using a buffer [50 mM HEPES (pH 7.4), 5 mM MgCl ₂ , 1 mM CaCl ₂ , 0.1% BSA] was added to each well. After adding 25 μL of a solution prepared so that the cell membrane fraction was 4 μL / well, 25 μL of a 0.2 nM ([ ¹²⁵ I] His ³ , MePhe ⁷ ) -NKB solution was added and mixed. After incubation at room temperature for 1 hour, suction filtration was performed using a GF / B filter pretreated with 0.3% polyethyleneimine (PEI) and washed with PEI washing buffer (50 mM HEPES, pH 7.4, 0.5% BSA). Subsequently, after washing with a washing buffer [50 mM HEPES (pH 7.4), 500 mM NaCl, 0.1% BSA], the filter was sufficiently dried. Thereafter, 25 μL of MicroScint O was added, and the radioactivity on the filter was measured at 1 min / well using TopCount NXT.

Test Example 2 Evaluation of NK3 Receptor Agonist Activity by Senktide Derivative of the Present Invention Changes in intracellular calcium concentration when Flp-In CHO cells forcibly expressing NK3 receptor were stimulated with various peptides were quantified.
First, NK3 receptor-expressing CHO cells were collected using trypsin and seeded in a 96-well plate at 4 × 10 ⁴ cells / well. After overnight culture at 37 ° C., 100 μL of calcium indicator was added, and further cultured for 1 hour. Subsequently, 25 μL of a peptide solution prepared by 5-fold concentration was added dropwise, and the time-dependent change in intracellular calcium concentration was observed for 60 seconds by FlexStation.

Test Example 3 Evaluation of NK1 receptor binding inhibitory activity of radioactive substance P (SP) with Sentidetide derivative of the present invention Flp-In CHO cells forcibly expressing NK1 receptor (Invitrogen's Flp-In CHO cells were treated with NK1 receptor) Using the receptor membrane fraction prepared from the gene introduced so as to stably express the peptide, the [ ¹²⁵ I] -BH-SP receptor binding inhibitory activity of various peptides was evaluated.
First, 50 μL of a compound prepared using a buffer [50 mM HEPES (pH 7.4), 5 mM MgCl ₂ , 1 mM CaCl ₂ , 0.1% BSA] was added to each well. After adding 25 μL of the solution prepared so that the cell membrane fraction was 4 μL / well, 25 μL of 0.2 nM [ ¹²⁵ I] -BH-SP solution was added and mixed. After incubation at room temperature for 1 hour, suction filtration was performed using a GF / B filter pretreated with 0.3% polyethyleneimine (PEI) and washed with PEI washing buffer [50 mM HEPES, pH 7.4, 0.5% BSA]. Subsequently, after washing with a washing buffer [50 mM HEPES (pH 7.4), 500 mM NaCl, 0.1% BSA], the filter was sufficiently dried. Thereafter, 25 μL of MicroScint O was added, and the radioactivity on the filter was measured at 1 min / well using TopCount NXT.

Test Example 4 Evaluation of NK2 Receptor Binding Inhibitory Activity of Radioactive NKA by Senktide Derivatives of the Present Invention Flp-In CHO Cells Forced to Express NK2 Receptor (In order to stably express NK2 receptor in Flp-In CHO cells of Invitrogen) Receptor membrane fractions prepared from the gene introduced into the gene were used to evaluate [ ¹²⁵ I] -NKA receptor binding inhibitory activity of various peptides.
First, 50 μL of a compound prepared using a buffer [50 mM HEPES (pH 7.4), 5 mM MgCl ₂ , 1 mM CaCl ₂ , 0.1% BSA] was added to each well. After adding 25 μL of the solution prepared so that the cell membrane fraction was 4 μL / well, 25 μL of 0.2 nM [ ¹²⁵ I] -NKA solution was added and mixed. After incubation at room temperature for 1 hour, the solution was suction filtered using a GF / B filter pretreated with 0.3% polyethyleneimine (PEI) and washed with PEI washing buffer [50 mM HEPES (pH 7.4), 0.5% BSA]. Subsequently, after washing with a washing buffer (50 mM HEPES (pH 7.4), 500 mM NaCl, 0.1% BSA), the filter was sufficiently dried. Thereafter, 25 μL of MicroScint O was added, and the radioactivity on the filter was measured at 1 min / well using TopCount NXT.

Test Example 5 Stability Evaluation of Senktide Derivatives of the Present Invention in Various Animal Serum and Porcine Hypothalamic Tissue Outer Fluid Under ice cooling, 4 μL of a peptide solution (10 mM) was added to and mixed with 196 μL of each animal tissue solution. Incubated at 37 ° C. Next, during the incubation, 30 μL of the evaluation solution was collected at each corresponding time. To the collected evaluation solution, 90 μL of MeCN was added under ice cooling to precipitate tissue protein. After shaking, the mixture was centrifuged (4 ° C., 13000 g, 10 min), and the resulting supernatant was analyzed by HPLC.

Test Example 6 Activity Evaluation of Senktide Derivatives of the Present Invention In Vivo Five ovariectomized goats were used, and the recording electrode was surgically placed in the vicinity of kisspeptin neurons in the hypothalamic arcuate nucleus. After a recovery period of about 1 month, nerve activity is measured in real time as an MUA in an awake goat, and it is confirmed that a transient increase in nerve activity (MUA volley) that lasts about 2 minutes occurs at a regular cycle. did. The MUA volley reflects the neural activity of kisspeptin neurons that secrete GnRH in pulses. In ovariectomized goats, the length of the endogenous MUA volley cycle is about 25 minutes (about ± 5 minutes), and varies somewhat depending on the individual, but within the same individual the cycle length is almost constant (within ± 2 minutes) ).
On the day of the test, the MUA measuring instrument was set and MUA was recorded continuously throughout the day. In addition, a jugular vein catheter for sample administration was placed. First, several endogenous MUA volleys occurring during the control time of about 2 hours were confirmed for each individual, and the average value (T) of the volley intervals (minutes) was calculated. Next, after the expression of an arbitrary MUA volley, the test sample solution was administered into the jugular vein at a timing of 1 / 2T. After administration, when MUA volley is expressed within 4 minutes, NK3R agonist activity is positive (◯), when it is expressed between 4 and 8 minutes, false positive (△), and when it occurs at more intervals (×) ). The MUA volley confirmed in the negative sample is endogenous.
The test sample solution was prepared as follows. On the test day, 7.5 μL of a test sample adjusted to a concentration of 10 mM in DMSO was dissolved in 4.5 mL of sterile distilled water. After confirming complete dissolution without precipitation, 0.5 mL of sterile 9% NaCl was added, and the final concentration of the solution was tested: 75 nmol / 5 mL, NaCl: 0.9%, DMSO: 0.15 %. 2 mL (30 nmol) was taken from this into a syringe and used for activity evaluation. Moreover, in principle, activity evaluation was performed using arbitrary 2 of 5 animals per test sample.

Table 9 shows the results of comparison of biological activity between Senktide and the Senktide derivative of the present invention based on Test Examples 1 and 6. As the Senktide derivative of the present invention, a tachykinin consensus sequence substituted was used.

Table 10 shows the results of comparison of biological activity between Senktide and the Senktide derivative of the present invention based on Test Examples 1 to 4 and 6. As the Senktide derivative of the present invention, the one obtained by converting MePhe, the third amino acid residue of Senktide, into a different amino acid was used.

Table 11 shows the results of comparison of biological activity between Senktide and the Senktide derivative of the present invention based on Test Examples 1 to 4 and 6. As the Senktide derivative of the present invention, one obtained by converting the N-terminus of Senktide was used.

Table 12 shows the results of comparison of biological activity between Senktide and the Senktide derivative of the present invention based on Test Examples 1 to 4 and 6. As the Senktide derivative of the present invention, the N-terminus of Senktide and one obtained by converting Asp, which is the first amino acid residue, to a different amino acid, or one having an amino acid introduced at the N-terminal side of the first amino acid residue were used. .

Table 13 shows the results of comparison of biological activity between Senktide and the Senktide derivative of the present invention based on Test Examples 1 to 4 and 6. As the Senktide derivative of the present invention, the one obtained by converting Asp, which is the N-terminal of Senktide and the first amino acid residue, into a different amino acid was used.

Tables 14 to 16 show the results of comparison of biological activity between Senktide and the Senktide derivative of the present invention based on Test Examples 1 to 4 and 6. As the Senktide derivative of the present invention, the one imitating the N-terminal structure (Succinyl-Asp) of Senktide was used.

Table 17 shows the results of comparison of biological activity between Senktide and the Senktide derivative of the present invention based on Test Examples 1 to 4. As the Senktide derivative of the present invention, a Senktide Gly-Leu converted to its peptide equivalent was used.

Based on Test Example 5, the stability of Senktide and the Senktide derivative of the present invention (E14) to neprilysin was compared. The results are shown in FIG.

This application is based on an application filed in Japan (Japanese Patent Application No. 2013-253534; filing date: December 6, 2013), the contents of which are incorporated in full herein.

[Sequence Listing Free Text]
SEQ ID NO: 1; Senktide
SEQ ID NO: 2; peptide 1
SEQ ID NO: 3; peptide 2
SEQ ID NO: 4; peptide 3
SEQ ID NO: 5; peptide 4
SEQ ID NO: 6; peptide 5
SEQ ID NO: 7; peptide 6
SEQ ID NO: 8; peptide 7
SEQ ID NO: 9; peptide 8
SEQ ID NO: 10; peptide 9
SEQ ID NO: 11; peptide 12
SEQ ID NO: 12; peptide 13
SEQ ID NO: 13; peptide 14
SEQ ID NO: 14; peptide 15
SEQ ID NO: 15; peptide 16
SEQ ID NO: 16; peptide 17
SEQ ID NO: 17; peptide 25
SEQ ID NO: 18; peptide 26
SEQ ID NO: 19; peptide 27
SEQ ID NO: 20; peptide 18
SEQ ID NO: 21; peptide 20
SEQ ID NO: 22; peptide 21
SEQ ID NO: 23; peptide 22
SEQ ID NO: 24; peptide 19
SEQ ID NO: 25; peptide 23
SEQ ID NO: 26; peptide 40b1, peptide 40b2
SEQ ID NO: 27; peptide 40c
SEQ ID NO: 28; peptide 40d1, peptide 40d2
SEQ ID NO: 29; peptide 40 g1, peptide 40 g2
SEQ ID NO: 30; peptide 111
SEQ ID NO: 31; peptide 112, peptide 113
SEQ ID NO: 32; peptide 39
SEQ ID NO: 33; peptide 114
SEQ ID NO: 34; peptide 101
SEQ ID NO: 35; peptide 102
SEQ ID NO: 36; peptide 35
SEQ ID NO: 37; peptide 36
SEQ ID NO: 38; peptide 105
SEQ ID NO: 39; peptide 31
SEQ ID NO: 40; peptide 109
SEQ ID NO: 41; peptide 28
SEQ ID NO: 42; peptide 32
SEQ ID NO: 43; peptide 110
SEQ ID NO: 44; peptide 116
SEQ ID NO: 45; peptide 117, peptide 118
SEQ ID NO: 46; peptide 119
SEQ ID NO: 47; peptide 120
SEQ ID NO: 48; peptide 121
SEQ ID NO: 49; peptide 122
SEQ ID NO: 50; peptide 123
SEQ ID NO: 51; peptide 124
SEQ ID NO: 52; peptide 125
SEQ ID NO: 53; peptide 126
SEQ ID NO: 54; peptide Y4a
SEQ ID NO: 55; peptide Y4b
SEQ ID NO: 56; peptide Y4c
SEQ ID NO: 57; peptide Y6a
SEQ ID NO: 58; peptide Y6b
SEQ ID NO: 59; peptide Y8
SEQ ID NO: 60; peptide Y10
SEQ ID NO: 61; peptide Y13
SEQ ID NO: 62; peptide E14
SEQ ID NO: 63; peptide E13
SEQ ID NO: 64; peptide E15
SEQ ID NO: 65; peptide E23
SEQ ID NO: 66; peptide E24

The highly active and highly selective NK3 receptor agonist obtained by the present invention is a therapeutic agent for diseases associated with excess or deficiency of tachykinins secretion, such as pain, sexual dysfunction associated with excess or deficiency of sex hormone secretion. It is useful as a drug for improvement and as a reagent for basic science experiments for developing therapeutics for these diseases.

Claims (11)

1. Formula (I)
   

   

   [Wherein X1 represents R1-A1 or a dicarboxylic acid residue;
   R1 is a hydrogen atom, acetyl group, oxalyl group, succinyl group, aminocarbonyl group, hydroxyaminocarbonyl group, hydrazinocarbonyl group, methoxydicarbonyl group, aminodicarbonyl group, aminosulfonyl group, N, N-dicarboxymethyl Represents aspartic acid or N-carboxymethylaspartic acid;
   A1 represents an L-aspartic acid, D-aspartic acid, L-glutamic acid, D-glutamic acid, or L-α-aminoadipic acid residue;
   A2 represents L-phenylalanine, L-tyrosine, L-aspartic acid, D-aspartic acid, L-glutamic acid, or D-glutamic acid residue;
   A3 represents LN-methylphenylalanine, LN-methylvaline, LN-methylisoleucine, LN-methyltyrosine, or LN-methyltryptophan residue;
   X4-X5 represents Gly-Leu or its dipeptide equivalent;
   Gly represents a glycine residue;
   Leu represents a leucine residue;
   Met-NH ₂ represents methionine amide] or a physiologically acceptable salt thereof.
2. X1 is
   

   

   Wherein R ^a and R ^{a ′} are the same or different and are a succinyl group or an oxalyl group; R ^b and R ^{b ′} are the same or different and are a succinyl group or an oxalyl group; R ^c and R ^{c ′} are the same Or a succinyl group or an oxalyl group; and R ^d and R ^{d ′} are the same or different and are a succinyl group or an oxalyl group).
   The compound according to claim 1 or a physiologically acceptable salt thereof.
3. The dipeptide equivalent is
   

   

   The compound according to claim 1 or 2, or a physiologically acceptable salt thereof.
4. The compound or physiologically acceptable salt thereof according to claim 1, wherein the compound or physiologically acceptable salt thereof is any one peptide represented by the following sequence or a physiologically acceptable salt: ;
   

   

   

   
5. The compound or physiologically acceptable salt thereof according to claim 1, wherein the compound or physiologically acceptable salt thereof is any one peptide represented by the following sequence or a physiologically acceptable salt: ;
   

   

   
6. A medicament comprising the compound according to claim 1 or a physiologically acceptable salt thereof.
7. The medicament according to claim 6, which is a neurokinin receptor agonist.
8. The medicament according to claim 7, wherein the neurokinin receptor is an NK3 receptor.
9. The medicament according to claim 6, wherein tachykinins and / or neurokinin receptors are therapeutic agents for diseases associated with their onset and progression.
10. The medicament according to claim 9, wherein the disease is sexual dysfunction associated with pain or excessive / deficiency of sex hormone secretion.
11. A reagent comprising the compound according to claim 1 or a physiologically acceptable salt thereof.