WO2022030580A1

WO2022030580A1 - Novel long-acting adrenomedullin derivative, method for producing same and pharmaceutical use thereof

Info

Publication number: WO2022030580A1
Application number: PCT/JP2021/029112
Authority: WO
Inventors: 和雄北村; 基生山▲崎▼
Original assignee: 国立大学法人宮崎大学
Priority date: 2020-08-06
Filing date: 2021-08-05
Publication date: 2022-02-10
Also published as: JPWO2022030580A1

Abstract

The present invention provides a novel long-acting adrenomedullin derivative that sustains the pharmacological effects of adrenomedullin, which is the parent compound thereof, and exhibits high biological stability when administered to a subject. One embodiment of the present invention pertains to a compound represented by formula (I) [in the formula: A represents a modifying group that is serum albumin; B represents a peptide moiety derived from adrenomedullin or a modified form thereof; and L represents a divalent linking group having a carboxyl group at at least one end thereof, wherein the α-amino group at the N-end of the peptide moiety B forms an amide bond together with the terminal carboxyl group of the linking group L so that the peptide moiety B is linked to the remainder], a salt thereof or a hydrate of the same. Another embodiment of the present invention pertains to: a method for producing the aforesaid compound, etc.; and a pharmaceutical and a prophylactic or therapeutic agent each comprising the compound, etc. as an active ingredient.　(I): A-L-B

Description

New long-acting adrenomedulin derivative, its manufacturing method and its pharmaceutical use

The present invention relates to a novel long-acting adrenomedullin derivative, a method for producing the same, and a pharmaceutical use thereof.

Adrenomedullin (hereinafter, also referred to as "AM") is a bioactive peptide isolated and identified from brown cell tissue in 1993 (Non-Patent Document 1). Initially discovered, AM was found to exert a strong vasodilatory antihypertensive effect. For example, Patent Document 1 describes a peptide having a blood pressure lowering effect, which comprises an amino acid sequence of human AM.

Subsequent studies have revealed that AM exerts various pharmacological actions such as cardiovascular protective action, anti-inflammatory action, angiogenic action and tissue repair promoting action. In addition, administration studies of AM to patients with various diseases have been conducted with the aim of applying the pharmacological action of AM to the treatment of diseases. In particular, the usefulness of AM as a therapeutic agent for inflammatory bowel disease, pulmonary hypertension, peripheral vascular disease or acute myocardial infarction is expected.

For example, Patent Document 2 describes an adrenomedullin or a derivative thereof having an activity of suppressing non-bacterial inflammation, or a salt thereof having an activity of suppressing non-bacterial inflammation as an active ingredient. Describes a prophylactic or therapeutic agent for non-bacterial inflammatory bowel disease contained as.

Patent Document 3 describes the method for preventing or treating an inflammatory bowel disease in a patient who requires the prevention or treatment of an inflammatory bowel disease in which the use of a steroid preparation, an immunosuppressive agent or a biological preparation is difficult or inadequate. The present invention comprises administering to the patient an effective amount of adrenomedulin, a derivative thereof having an activity of suppressing inflammation, or a salt of the adrenomedulin or the derivative thereof having an activity of suppressing inflammation. The prevention or treatment method is described.

Since AM is a peptide, it has a short half-life in vivo due to a metabolic reaction in vivo (for example, in blood). Therefore, when administering AM to a subject, it is necessary to select a continuous administration method such as continuous intravenous infusion. In addition, AM has a strong vasodilatory action in addition to a pharmacological action such as a cardiovascular protective action, an anti-inflammatory action, an angiogenesis action and a tissue repair promoting action. Therefore, when AM is administered to a subject, it may cause unwanted side effects such as excessive blood pressure reduction due to its strong vasodilatory effect. In view of these issues, long-lasting adrenomedullin derivatives have been developed that can substantially suppress unwanted side effects while maintaining the pharmacological action of adrenomedullin (Patent Documents 4 to 6).

Further, as a technique for improving pharmacokinetics, a conjugate of a drug and serum albumin is known (Patent Documents 7 and 8).

Japanese Patent No. 2774769 Japanese Patent No. 4830093 Japanese Patent No. 5954736 International Publication No. 2015/14 1919 International Publication No. 2017/047788 International Publication No. 2018/181638 Japanese Patent No. 5599822 Japanese Patent No. 5639039

As described above, various adrenomedulin derivatives have been developed in order to improve the pharmacokinetics of adrenomedullin from the viewpoint of improving sustainability in vivo. However, there was room for improvement in known adrenomedullin derivatives. Adrenomedullin has a strong vasodilatory action in addition to pharmacological actions such as cardiovascular protective action, anti-inflammatory action, angiogenic action and tissue repair promoting action. Therefore, when adrenomedullin or a derivative thereof is administered to a subject, it is not desirable such as excessive decrease in blood pressure due to a strong vasodilatory effect, tachycardia and / or increase in renin activity associated with an increase in reflex sympathetic nerve activity. May cause side effects. The occurrence of such side effects can be problematic, especially when adrenomedullin or its derivatives are used with the expectation that it will exhibit pharmacological effects other than vasodilatory effects. In order to avoid the above-mentioned problems, a drug containing adrenomedullin or a derivative thereof as an active ingredient is administered to a subject by continuous intravenous infusion at a dose that substantially does not cause undesired side effects. Needed to be done. Such a method of administration may impose a burden on the subject.

The adrenomedulin derivative, which maintains the pharmacological action of adrenomedullin and has improved persistence in vivo, exhibits the pharmacological effect of adrenomedullin even when administered once to a subject, without causing unwanted side effects. Expected to be possible. Therefore, it is an object of the present invention to provide a novel long-acting adrenomedullin derivative that exhibits high biological stability when administered to a subject while maintaining the pharmacological action of the parent compound adrenomedullin. ..

The present inventors have examined various means for solving the above-mentioned problems. By linking the N-terminal α-amino group of adrenomedullin and serum albumin via a divalent linking group, the present inventors maintain the same biological activity as the parent compound adrenomedullin, and adrenomedullin. We have found that they have significantly better pharmacokinetics, for example with respect to biological stability. The present inventors have completed the present invention based on the above findings.

That is, the present invention includes the following aspects and embodiments.
(1) Equation (I):
ALB (I)
[During the ceremony,
A is a modifying group that is serum albumin,
B is a peptide moiety derived from adrenomedulin or a variant thereof.
L is a divalent linking group having a carboxyl group at at least one end.
The peptide moiety B is linked to the rest by the N-terminal α-amino group forming an amide bond with the carboxyl group at the end of the linking group L. ]
A compound represented by, or a salt thereof, or a hydrate thereof.
(2) The compound according to the above embodiment (1), a salt thereof, or a hydrate thereof, wherein the modifying group A is human serum albumin consisting of the amino acid sequence of SEQ ID NO: 14.
(3) The linking group L is a substituted or unsubstituted divalent hydrocarbon group (the divalent hydrocarbon group is one or more complex atoms, a heterocycle, an amide group (-CO-NH-), an ester. The compound according to the above embodiment (1) or (2) or a salt thereof, which is a group (-CO-O-) or a urethane group (-O-CO-NH-) may be contained), or a salt thereof. Hydrocarbon.
(4) The linking group L has an ethylene oxide unit having a number of repetitions in the range of 2 to 24, and has one or more complex atoms, a heterocycle, an amide group (-CO-NH-), and an ester group (-CO). -O-), or a divalent hydrocarbon group which may contain a urethane group (-O-CO-NH-), the compound according to embodiment (3) or a salt thereof, or a hydrate thereof. ..
(5) Adrenomedullin or its modified product is as follows:
(I) A peptide consisting of the amino acid sequence of adrenomedulin,
(Ii) A peptide consisting of the amino acid sequence of adrenomedullin, in which two cysteine residues in the amino acid sequence form a disulfide bond.
(Iii) In the peptide of (ii), the peptide in which the disulfide bond is substituted with an ethylene group,
(Iv) A peptide in which 1 to 15 amino acid residues are deleted, substituted or added in any of the peptides of (i) to (iii).
(V) In any of the peptides (i) to (iv), the peptide in which the C-terminal is amidated, and in any of the peptides (vi), (i) to (iv), the glycine residue at the C-terminal remains. The compound according to any one of the above-described embodiments (1) to (4), a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of peptides to which a group is added.
(6) Adrenomedullin or its modified product is as follows:
(I) A peptide consisting of the amino acid sequence of adrenomedulin,
(Ii) A peptide consisting of the amino acid sequence of adrenomedullin, in which two cysteine residues in the amino acid sequence form a disulfide bond.
(V) In the peptide of (i) or (ii), the peptide in which the C-terminal is amidated, and in the peptide of (vi) (i) or (ii), a glycine residue is added to the C-terminal. The compound according to the above-mentioned embodiment (5), a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of peptides.
(7) Adrenomedullin or its modified product is as follows:
(Iv') In any of the peptides (i) to (iii), a peptide in which the amino acid residues at positions 1 to 15, 1 to 10 or 1 to 5 from the N-terminal side are deleted,
(V) Select from the group consisting of the peptide in which the C-terminal is amidated in the peptide of (iv') and the peptide in which the glycine residue is added to the C-terminal in the peptide of (vi) (iv'). The peptide according to the above embodiment (5), a salt thereof, or a hydrate thereof.
(8) Adrenomedullin or a modified product thereof is as follows:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(B) A peptide consisting of the amino acid sequence of SEQ ID NO: 4, or a peptide consisting of the amino acid sequence of SEQ ID NO: 4 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(C) A peptide consisting of the amino acid sequence of SEQ ID NO: 6 or a peptide consisting of the amino acid sequence of SEQ ID NO: 6 in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(D) A peptide consisting of the amino acid sequence of SEQ ID NO: 8 or a peptide consisting of the amino acid sequence of SEQ ID NO: 8 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(E) A peptide consisting of the amino acid sequence of SEQ ID NO: 10 or a peptide consisting of the amino acid sequence of SEQ ID NO: 10 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(F) A peptide consisting of the amino acid sequence of SEQ ID NO: 12, or a peptide consisting of the amino acid sequence of SEQ ID NO: 12 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(G) In any of the peptides (a) to (f), the peptide in which the disulfide bond is substituted with an ethylene group;
(H) A peptide in which 1 to 15 amino acid residues are deleted, substituted or added in any of the peptides (a) to (g);
(I) A peptide in which the C-terminal is amidated in any of the peptides (a) to (h); and a glycine residue at the C-terminal in any of the peptides (j) (a) to (h). Peptides with added groups;
The compound according to any one of the above-described embodiments (1) to (5), a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of.
(9) Adrenomedullin or its modified product is as follows:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(B) A peptide consisting of the amino acid sequence of SEQ ID NO: 4, or a peptide consisting of the amino acid sequence of SEQ ID NO: 4 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(C) A peptide consisting of the amino acid sequence of SEQ ID NO: 6 or a peptide consisting of the amino acid sequence of SEQ ID NO: 6 in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(D) A peptide consisting of the amino acid sequence of SEQ ID NO: 8 or a peptide consisting of the amino acid sequence of SEQ ID NO: 8 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(E) A peptide consisting of the amino acid sequence of SEQ ID NO: 10 or a peptide consisting of the amino acid sequence of SEQ ID NO: 10 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(F) A peptide consisting of the amino acid sequence of SEQ ID NO: 12, or a peptide consisting of the amino acid sequence of SEQ ID NO: 12 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(I) A peptide in which the C-terminal is amidated in any of the peptides (a) to (f); and a glycine residue at the C-terminal in any of the peptides (j) (a) to (f). Peptides with added groups;
The compound according to the above-mentioned embodiment (8), a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of.
(10) Adrenomedullin or a modified product thereof is as follows:
(H') In any of the peptides (a) to (d), the amino acid residues at positions 1 to 15, 1 to 10 or 1 to 5 from the N-terminal side are deleted, or ( In the peptide of e) or (f), the peptide in which the amino acid residues at positions 1 to 13, 1 to 8 or 1 to 5 are deleted from the N-terminal side;
(I) In the peptide of (h'), the peptide in which the C-terminal is amidated; and in the peptide of (j) (h'), the peptide in which the glycine residue is added to the C-terminal;
The compound according to the above-mentioned embodiment (8), a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of.
(11) Between the precursor of modifying group A, which is serum albumin, and the precursor of divalent linking group L, and between the precursor of peptide moiety B derived from adrenomedulin or its modification, and the divalent linking group L. The compound according to any one of the above-described embodiments (1) to (10), which comprises a linking step of linking at least one of the precursors of the above to obtain a compound represented by the formula (I). A method for producing the salt or its hydrate.
(12) A drug containing the compound according to any one of embodiments (1) to (10), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable hydrate thereof as an active ingredient.
(13) Heart failure, acute myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Bechet's disease, diabetes, diabetic The pharmaceutical according to embodiment (12) above, for use in the prevention or treatment of nephropathy, diabetic retinopathy, pulmonary fibrosis, sepsis or septic shock.
(14) Heart failure, acute, containing the compound according to any one of the above embodiments (1) to (10), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable hydrate thereof as an active ingredient. Myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Bechet's disease, diabetes, diabetic nephropathy, diabetic retina A prophylactic or therapeutic agent for diabetes, pulmonary fibrosis, arrhythmia or arrhythmic shock.
(15) The compound according to any one of the above embodiments (1) to (10), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable hydrate thereof, and one or more pharmaceutically acceptable salts. A pharmaceutical composition containing a carrier.
(16) Heart failure, acute myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Bechet's disease, diabetes, diabetic The pharmaceutical composition according to embodiment (15), wherein the pharmaceutical composition is used for the prevention or treatment of nephropathy, diabetic retinopathy, pulmonary fibrosis, sepsis or septic shock.
(17) Prevention or treatment of one or more symptoms, diseases and / or disorders selected from the group consisting of cardiovascular disease, brain / neurological disease, digestive system disease, endocrine and metabolic disease, respiratory system disease and other diseases. Administering an effective amount of the compound according to any one of the above-mentioned embodiments (1) to (10), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof to a required subject. A method for preventing or treating the above-mentioned symptoms, diseases and / or disorders.
(18) One or more symptoms, diseases and / or disorders include heart failure, acute myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcer. The method according to embodiment (17), wherein the method is sexual colitis, intestinal Bechette's disease, diabetes, diabetic nephropathy, diabetic retinopathy, pulmonary fibrosis, septicemia or septic shock.
(19) The compound according to any one of the above-described embodiments (1) to (10) or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of one or more symptoms, diseases and / or disorders. Those pharmaceutically acceptable solvates.
(20) One or more symptoms, diseases and / or disorders include heart failure, acute myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcer. The compound for use according to the embodiment (19) or pharmaceutical thereof, which is sexual colitis, intestinal Bechette's disease, diabetes, diabetic nephropathy, diabetic retinopathy, pulmonary fibrosis, sepsis or septic shock. Acceptable salts, or their pharmaceutically acceptable arrhythmias.
(21) The compound according to any one of the above embodiments (1) to (10) or a pharmaceutically acceptable salt thereof in the manufacture of a pharmaceutical agent for the prevention or treatment of one or more symptoms, diseases and / or disorders. , Or the use of their pharmaceutically acceptable solvates.
(22) The compound according to any one of the above-described embodiments (1) to (10), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof for the prevention or treatment of one or more symptoms, diseases and / or disorders. Use of pharmaceutically acceptable solvates.
(23) One or more symptoms, diseases and / or disorders include heart failure, acute myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcer. The use according to embodiment (21) or (22) above, which is sexual colitis, intestinal Bechette's disease, diabetes, diabetic nephropathy, diabetic retinopathy, pulmonary fibrosis, sepsis or septic shock.

Each aspect of the present invention makes it possible to provide a novel long-acting adrenomedullin derivative that exhibits high biological stability when administered to a subject while maintaining the pharmacological action of the parent compound adrenomedullin. ..

This specification includes the contents described in the specification and / or the drawings of Japanese Patent Application No. 2020-133777, which is the basis of the priority of the present application.

FIG. 1 shows the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting (WB) of the synthesized novel adrenomedullin derivative (hSA-AM) in Experiment I. In the figure, A shows the gel stained with Coomassie Brilliant Blue after SDS-PAGE, and B shows the membrane obtained by WB. In A and B, M represents a molecular weight marker (Precision Plus Protein Dual Xtra Standards), lane 1 represents 68 ng of hSA, and lane 2 represents 180 ng of hSA-AM. FIG. 2 shows a UV chromatogram with a wavelength of 280 nm by gel filtration chromatography (GF) of the novel adrenomedullin derivative (hSA-AM) synthesized in Experiment I. In the figure, A is a chromatogram of 30 μg hSA-AM, B is a chromatogram of 30 μg hSA, and C is a chromatogram of purified hSA-AM. In the figure, the horizontal axis is the retention time (minutes) of the reverse phase HPLC, and the vertical axis is the UV absorption intensity (mV) at a wavelength of 280 nm. FIG. 3 shows the dose-response curve of intracellular cAMP production in HEK293 cells to the added concentration of native adrenomedullin (natural AM) or novel adrenomedullin derivative (hSA-AM) in Experiment II-1. In the figure, the black circles (●) are the average values of the experiments to which natural AM was added, the white squares (□) are the average values of the experiments to which hSA-AM was added, and the error bars represent SEM. In the figure, the horizontal axis is the logarithm (M) of the added concentration of natural AM or hSA-AM, and the vertical axis is the intracellular cAMP production amount (fmol) in HEK293 cells that stably express the AM1 receptor. / Well). FIG. 4 shows the time course of the amount of natural adrenomedullin (natural AM) or novel adrenomedullin derivative (hSA-AM) in rat plasma after subcutaneous administration in Experiment II-2. In the figure, A is the result of natural AM and B is the result of hSA-AM. In the figure, the horizontal axis is the elapsed time (A: time, B: day) after subcutaneous administration, and the vertical axis is the plasma peptide concentration (pM) after administration of natural AM or hSA-AM. .. FIG. 5 shows the effect of a single subcutaneous administration of an AM derivative (hSA-AM) on spontaneously hypertensive rats (SHR) in Experiment II-3 to suppress the increase in blood pressure. In the figure, A is the time course of systolic blood pressure (sBP), and B is the time course of diastolic blood pressure (dBP). The horizontal axis is the elapsed time (days), and the vertical axis shows the difference in blood pressure (A: difference in sBP, B: difference in dBP) (mmHg) obtained by subtracting the blood pressure before administration from the blood pressure on each measurement day. .. The values in the figure are the average values of each group, and the error bars represent SEM. FIG. 6 shows changes in the disease activity index (DAI) score due to subcutaneous administration of the AM derivative (hSA-AM) to dextran sodium sulfate (DSS) -induced colitis model mice in Experiment II-4. In the figure, the horizontal axis is the elapsed time (day), and the vertical axis is the DAI score. The values in the figure are the average values of each group, and the error bars represent SEM.

<1. Adrenomedullin derivative>
Adrenomedullin (AM) is a bioactive peptide isolated and identified from human brown cell tissue in 1993 (SEQ ID NO: 1, Non-Patent Document 1). A peptide consisting of the amino acid sequence of SEQ ID NO: 1, the C-terminal is amidated, and the two cysteine residues at positions 16 and 21 in the amino acid sequence form a disulfide bond is a mature natural peptide. Corresponds to type human adrenomedulin (hereinafter, also referred to as "hAM (1-52)"). The peptide consisting of the amino acid sequence of SEQ ID NO: 1 corresponds to the pre-translational (ie immature) form of natural human adrenomedullin that has undergone post-translational modification of C-terminal amidation and disulfide of cysteine residues.
hAM (1-52) YRQSMNNFQGLRSFGCRFGTC-TVQKLAHQIYQFTDKDKDNVA-PRSKISPQGY-CONH ₂ (SEQ ID NO: 1)

At the time of discovery, AM was found to exert a strong vasodilatory antihypertensive effect. Subsequent studies have revealed that AM exerts a variety of pharmacological actions such as cardiovascular protective action, anti-inflammatory action, angiogenic action and tissue repair promoting action. In addition, administration studies of AM to patients with various diseases have been conducted with the aim of applying the pharmacological action of AM to the treatment of diseases.

Since AM is a peptide, it has a short half-life in vivo due to a metabolic reaction in vivo (for example, in blood). In addition, AM has a strong vasodilatory action in addition to a pharmacological action such as a cardiovascular protective action, an anti-inflammatory action, an angiogenesis action and a tissue repair promoting action. Therefore, when AM is administered to subjects, it causes undesired side effects such as excessive decrease in blood pressure due to strong vasodilatory action, tachycardia associated with increased reflex sympathetic nerve activity, and / or increased renin activity. there is a possibility. The occurrence of such side effects can be problematic, especially when AM or its derivatives are used with the expectation that they will exhibit pharmacological effects other than vasodilatory effects. In order to avoid the above-mentioned problems, the prior art drug containing AM or its derivative as an active ingredient is administered to the subject by continuous intravenous infusion at a dose that substantially does not cause undesired side effects. Needed to be done. Such a method of administration may impose a burden on the subject.

By linking the N-terminal α-amino group of AM and serum albumin via a divalent linking group, the present inventors maintain the same biological activity as the parent compound AM, and AM. We have found that they have significantly better pharmacokinetics, for example with respect to biological stability.

Therefore, one aspect of the present invention is the formula (I) :.
ALB (I)
The present invention relates to a compound represented by the above, a salt thereof, or a hydrate thereof. In the present specification, the compound represented by the formula (I) or a salt thereof, or a solvate thereof may be referred to as an "adrenomedullin derivative" or an "AM derivative".

In each aspect of the invention, AM is not only a human-derived peptide (SEQ ID NO: 1, Non-Patent Document 1), but also, for example, pig (SEQ ID NO: 4), dog (SEQ ID NO: 6), bovine (SEQ ID NO: 8), and the like. It may be a peptide (ortholog) derived from another non-human mammal (eg, warm-blooded animal) such as rat (SEQ ID NO: 10) or mouse (SEQ ID NO: 12). In vivo, these peptides have two cysteine residues in their amino acid sequence forming a disulfide bond and the C-terminus being amidated. In the present specification, the peptide having a disulfide bond and a C-terminal amide group may be referred to as "natural adrenomedullin" or simply "adrenomedullin". Each aspect of the present invention can be applied to any of the above peptides.

As used herein, "C-terminal amidation" means an aspect of post-translational modification of a peptide in vivo, and specifically, the main chain carboxyl group of the C-terminal amino acid residue of the peptide is an amide group. It means a reaction that is transformed into a form. Further, in the present specification, "formation of a disulfide bond of a cysteine residue" or "disulfide formation of a cysteine residue" means one aspect of post-translational modification of a peptide in vivo, and specifically, the peptide. It means a reaction in which two cysteine residues in an amino acid sequence form a disulfide bond (-SS-). Many bioactive peptides produced in vivo are initially biosynthesized as higher molecular weight precursor proteins, such as C-terminal amidation and / or cysteine residue disulfide during intracellular translocation. After translation, it undergoes a modification reaction to become a mature physiologically active peptide. C-terminal amidation usually proceeds by the action of C-terminal amyloid enzyme on precursor protein. In the case of a physiologically active peptide having a C-terminal amide group, in the precursor protein, a glycine (Gly) residue is bound to the C-terminal carboxyl group to be amidated, and the Gly residue is a C-terminal amidating enzyme. Is converted to a C-terminal amide group. In addition, the C-terminal propeptide of precursor protein contains a repeating sequence of a combination of basic amino acid residues such as Lys-Arg or Arg-Arg (Mizuno, Biochemistry Vol. 61, No. 12, No. 12). Pp. 1435-1461 (1989)). Disulfide formation of cysteine residues can proceed under oxidative conditions. In vivo, disulfide formation of cysteine residues usually proceeds by the action of protein disulfide isomerase on the precursor protein.

In formula (I), B is a peptide moiety derived from adrenomedullin or a variant thereof. Peptide moiety B preferably has adrenomedulin activity by itself. That is, the peptide moiety B is preferably an adrenomedullin or a peptide moiety derived from a modified form thereof having adrenomedulin activity. In each aspect of the invention, the "peptide moiety derived from adrenomedulin or a modification thereof" is one hydrogen atom from AM or a modification thereof (usually one hydrogen atom of an amino group, typically one hydrogen atom). It means a monovalent free group having a structure in which one hydrogen atom of the α-amino group of the amino acid residue at the N-terminal is removed. In each aspect of the invention, "modified adrenomedullin" means a peptide chemically modified from the native AM described above. Further, in the present invention, "adrenomedullin activity" means the biological activity of AM. Examples of the adrenomedullin activity include the following.

(1) Cardiovascular system: vasodilatory effect, blood pressure lowering effect, blood pressure increase inhibitory effect, heart rate increase / heart failure improving effect, pulmonary hypertension improving effect, angiogenesis effect, lymphangiogenesis effect, vascular endothelial function improving effect , Vascular permeability control, Endothelial cell adhesion control, Endothelial barrier protective action, Anti-arteriosclerotic action, Myocardial protective action (eg, Myocardial protective action in ischemia-reperfusion injury or inflammation), Remodeling inhibitory action after myocardial infarction , Cardiac hypertrophy inhibitory action, and andothetensin converting enzyme inhibitory action.
(2) Kidney / water electrolyte system: diuretic effect, natriuretic effect, antidiuretic hormone inhibitory effect, aldosterone lowering effect, renal protective effect (for example, myocardial protective effect in hypertension or ischemia-reperfusion injury), diabetic nephropathy suppression Action, C3 nephropathy suppressing action, drinking behavior suppressing action, and salt requirement suppressing action.
(3) Brain / nervous system: neuroprotective / cerebral disorder inhibitory action, anti-inflammatory action, apoptosis inhibitory action (for example, ischemia-reperfusion injury or apoptosis inhibitory action in inflammation), autoregulatory ability maintenance action, oxidative stress suppressive action, Dementia improving effect and sympathetic depressant effect.
(4) Genitourinary system: erection improving action, blood flow improving action, and implantation promoting action.
(5) Digestive system: anti-ulcer action, tissue repair action, mucosal neoplastic action, intestinal barrier protection action, blood flow improving action, anti-inflammatory action, and liver function improving action.
(6) Orthopedic system: Osteoblast stimulating action and arthritis improving action.
(7) Endocrine metabolism system: adipocyte differentiation effect, lipolysis control effect, insulin sensitivity improving effect, insulin secretion control effect, antidiuretic hormone secretion inhibitory effect, and aldosterone secretion inhibitory effect.
(8) Respiratory system: Bronchial dilation effect, lung protection effect, emphysema improving effect, pulmonary fibrosis suppression, pneumonia suppression, bronchitis suppression effect, and respiratory improvement effect.
(9) Immune system: C3b degradation promoting action.
(10) Others: Circulation improving action, anti-inflammatory action, cytokine control action, organ protection action, oxidative stress suppressing action, tissue repair action (for example, anti-decubitus action), sepsis improving action, septic shock improving action, many Suppressive action of organ failure, suppressive action of autoimmune disease, suppressive action of diabetic retinopathy, antibacterial action, hair growth action, and hair nourishing action.

The blood pressure lowering action is preferably a vasodilatory antihypertensive action. The anti-inflammatory action in the digestive system is a preventive or therapeutic action for inflammatory bowel diseases such as steroid-resistant or steroid-dependent inflammatory bowel diseases (eg, ulcerative colitis, Crohn's disease or intestinal Behcet's disease). It is preferable to have.

The adrenomedulin activity exemplified above expressed by AM is usually expressed through an increase in the concentration of intracellular cAMP. Therefore, an increase in the concentration of intracellular cAMP can be used as an index of the adrenomedullin activity of the compound of this embodiment. In each aspect of the present invention, the action of increasing the intracellular cAMP concentration is, for example, by adding a target compound to a cultured cell line (HEK293 cell line) in which AM type 1 receptor (AM1 receptor) is stably expressed, and cells are used. It can be evaluated by measuring the production amount of intracellular cAMP. The compound of this embodiment has an action of increasing the concentration of intracellular cAMP substantially equivalent to that of natural AM. Therefore, the compound of this embodiment can express biological activity (that is, adrenomedullin activity) substantially equivalent to that of natural AM through an increase in the concentration of intracellular cAMP.

Adrenomedullin or its modifications are listed below:
(I) A peptide consisting of the amino acid sequence of adrenomedulin,
(Ii) A peptide consisting of the amino acid sequence of adrenomedullin, in which two cysteine residues in the amino acid sequence form a disulfide bond.
(Iii) In the peptide of (ii), the peptide in which the disulfide bond is substituted with an ethylene group,
(Iv) A peptide in which 1 to 15 amino acid residues are deleted, substituted or added in any of the peptides of (i) to (iii).
(V) In any of the peptides (i) to (iv), the peptide in which the C-terminal is amidated, and in any of the peptides (vi), (i) to (iv), the glycine residue at the C-terminal remains. It is preferably a peptide selected from the group consisting of peptides to which a group is added.

In one embodiment, the adrenomedullin or a modification thereof is as follows:
(I) A peptide consisting of the amino acid sequence of adrenomedulin,
(Ii) A peptide consisting of the amino acid sequence of adrenomedullin, in which two cysteine residues in the amino acid sequence form a disulfide bond.
(V) In the peptide of (i) or (ii), the peptide whose C-terminal is amidated, and in the peptide of (vi) (i) or (ii), a glycine residue is added to the C-terminal. It is more preferable that the peptide is selected from the group consisting of peptides.

In the peptides (i) to (vi), the peptide consists of the amino acid sequence of AM contained in (v), the C-terminal is amidated, and the two cysteine residues in the amino acid sequence are disulfides. The peptide forming the bond corresponds to mature native AM. The peptide consisting of the amino acid sequence of AM in (i) corresponds to the pre-translational modification (ie, immature) form of native AM with C-terminal amidation and disulfide of cysteine residues. In the peptides (i) to (vi), the peptides other than the peptides described above correspond to modified versions of AM.

The peptide (ii) is formed by air-oxidizing the thiol groups of the two cysteine residues of the peptide (i) or by oxidizing them with an appropriate oxidizing agent to convert them into disulfide bonds. Can be made to. By using the peptide of (ii) above, the three-dimensional structure of the peptide portion B can be made to resemble the three-dimensional structure of the natural AM. Thereby, the adrenomedullin activity of the compound represented by the formula (I) can be made substantially equivalent to that of the natural AM.

The peptide of (iii) can be formed by converting the disulfide bond of the peptide of (ii) into an ethylene group. Substitution of disulfide bonds to ethylene groups can be performed by methods well known in the art (O. Keller et al., Helv. Chim. Acta, 1974, Vol. 57, p. 1253). By using the peptide of (iii) above, the three-dimensional structure of the peptide portion B can be stabilized. Thereby, the compound represented by the formula (I) can continuously express the adrenomedulin activity in the living body.

In the peptide (iv), the number of amino acid residues deleted, substituted or added is preferably in the range of 1 to 15, more preferably in the range of 1 to 10, and 1 to 8. The range of 1 is more preferable, the range of 1 to 5 is particularly preferable, and the range of 1 to 3 is most preferable. Suitable peptides (iv) are 1 to 15 positions, 1 to 12 positions, 1 to 10 positions, 1 to 8 positions, and 1 to 5 positions from the N-terminal side in any of the peptides (i) to (iii). A peptide in which the amino acid residue at the position or the 1st to 3rd position is deleted, and the more preferable peptide (iv) is the peptide of any of (i) to (iii), 1 to 1 to 1 from the N-terminal side. It is a peptide in which amino acid residues at

positions

15, 1 to 10 or 1 to 5 are deleted. In the preferred peptide, one or more (eg, 1-5, 1-3, or 1 or 2) amino acid residues may be further deleted, substituted or added. By using the peptide of (iv) above, the adrenomedullin activity of the compound represented by the formula (I) can be made substantially equivalent to that of natural AM. In addition, by using the peptide of (iv) above, the compound represented by the formula (I) can continuously express adrenomedulin activity in vivo.

The peptide of (vi) can be converted into the peptide of (v) by converting the C-terminal glycine residue into a C-terminal amide group by the action of the C-terminal amidating enzyme. Therefore, by administering the peptide (vi) to a subject, a C-terminal amidated peptide can be formed in the living body of the subject after a lapse of a certain period of time. Thereby, the compound represented by the formula (I) can continuously express the adrenomedulin activity in the living body.

Adrenomedullin or its modifications are listed below:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(B) A peptide consisting of the amino acid sequence of SEQ ID NO: 4, or a peptide consisting of the amino acid sequence of SEQ ID NO: 4 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(C) A peptide consisting of the amino acid sequence of SEQ ID NO: 6 or a peptide consisting of the amino acid sequence of SEQ ID NO: 6 in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(D) A peptide consisting of the amino acid sequence of SEQ ID NO: 8 or a peptide consisting of the amino acid sequence of SEQ ID NO: 8 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(E) A peptide consisting of the amino acid sequence of SEQ ID NO: 10 or a peptide consisting of the amino acid sequence of SEQ ID NO: 10 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(F) A peptide consisting of the amino acid sequence of SEQ ID NO: 12, or a peptide consisting of the amino acid sequence of SEQ ID NO: 12 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(G) In any of the peptides (a) to (f), the peptide in which the disulfide bond is substituted with an ethylene group;
(H) A peptide in which 1 to 15 amino acid residues are deleted, substituted or added in any of the peptides (a) to (g);
(I) A peptide in which the C-terminal is amidated in any of the peptides (a) to (h); and a glycine residue at the C-terminal in any of the peptides (j) (a) to (h). Peptides with added groups;
It is more preferable that the peptide is selected from the group consisting of.

Adrenomedullin or its modifications are listed below:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(B) A peptide consisting of the amino acid sequence of SEQ ID NO: 4, or a peptide consisting of the amino acid sequence of SEQ ID NO: 4 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(C) A peptide consisting of the amino acid sequence of SEQ ID NO: 6 or a peptide consisting of the amino acid sequence of SEQ ID NO: 6 in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(D) A peptide consisting of the amino acid sequence of SEQ ID NO: 8 or a peptide consisting of the amino acid sequence of SEQ ID NO: 8 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(E) A peptide consisting of the amino acid sequence of SEQ ID NO: 10 or a peptide consisting of the amino acid sequence of SEQ ID NO: 10 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(F) A peptide consisting of the amino acid sequence of SEQ ID NO: 12, or a peptide consisting of the amino acid sequence of SEQ ID NO: 12 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(I) A peptide in which the C-terminal is amidated in any of the peptides (a) to (f); and a glycine residue at the C-terminal in any of the peptides (j) (a) to (f). Peptides with added groups;
It is more preferable that the peptide is selected from the group consisting of.

In one embodiment, the adrenomedullin or a modification thereof is as follows:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(I) In the peptide of (a), the peptide in which the C-terminal is amidated; and in the peptide of (j) (a), the peptide in which the glycine residue is added to the C-terminal;
It is preferably a peptide selected from the group consisting of.

In the peptide (h), the number of amino acid residues deleted, substituted or added is preferably in the range of 1 to 12, more preferably in the range of 1 to 10, and is preferably in the range of 1 to 8. The range of 1 is more preferable, the range of 1 to 5 is particularly preferable, and the range of 1 to 3 is most preferable. Suitable peptides (h) are 1 to 15 positions, 1 to 12 positions, 1 to 10 positions, 1 to 8 positions, and 1 to 5 positions from the N-terminal side in any of the peptides (a) to (g). Peptides in which the amino acids at

positions

1 or 1 to 3 are deleted, and the more preferable peptide (h) is the peptide at positions 1 to 15 from the N-terminal side in any of the peptides (a) to (d). , 1 to 10 or 1 to 5 amino acid residues are deleted, or in the peptide of (e) or (f), 1 to 13 positions, 1 to 8 positions or 1 to 1 to N-terminal side. It is a peptide in which the amino acid residue at position 5 is deleted. In the preferred peptide, one or more amino acids (eg, 1-5, 1-3, or 1 or 2) may be further deleted, substituted or added. By using the peptide of (h), the adrenomedullin activity of the compound represented by the formula (I) can be made substantially equivalent to that of natural AM. Further, by using the peptide of the above (h), the compound represented by the formula (I) can continuously express the adrenomedullin activity in the living body.

The peptides exemplified above as adrenomedulin or a modified form thereof usually have adrenomedulin activity. In the peptides exemplified above, the peptide having the above-mentioned characteristics and adrenomedullin activity is, for example, biological as compared with AM while maintaining substantially the same pharmacological action as the parent compound AM. It can have significantly better pharmacokinetics with respect to stability.

In formula (I), A is a modifying group that is serum albumin. Serum albumin is the major protein in mammalian blood. Serum albumin is involved in the transport and metabolism of various substances in the living body. For example, serum albumin recognizes and binds to endothelial cell gp60. As a result, serum albumin is known to have excellent migration activity on endothelial cells. Therefore, the conjugate of the drug and serum albumin is known as a technique for improving pharmacokinetics (for example, Japanese Patent No. 5959822 (Patent Document 7) and Japanese Patent No. 5639039 (Patent Document 8)). Therefore, the compound represented by the formula (I) of this embodiment having the modifying group A which is serum albumin has significantly superior pharmacokinetics, for example, in terms of biological stability as compared with the native AM. be able to.

In the formula (I), the mammal from which the serum albumin used as the modifying group A is derived is a drug containing the compound represented by the formula (I) of one aspect of the present invention as an active ingredient, which is described below. It can be selected as appropriate based on the target to be applied. Modifying group A is serum albumin from human or non-human mammals (eg, warm-blooded animals such as pigs, dogs, cows, rats, mice, guinea pigs, rabbits, chickens, sheep, cats, monkeys, hamadryas baboons or chimpanzees). It is preferable that the serum albumin is derived from the same human or non-human mammal as the subject to which the medicine of one aspect of the present invention is applied. In particular, the modifying group A is preferably human serum albumin consisting of the amino acid sequence of SEQ ID NO: 14. Human serum albumin (hereinafter, also referred to as “hSA”) consisting of the amino acid sequence of SEQ ID NO: 14 is a protein consisting of 585 amino acid residues having a molecular weight of about 66,000 Da. By using serum albumin derived from the human or non-human mammal as the modifying group A, the compound represented by the formula (I) of this embodiment can maintain the pharmacological action of the natural AM while maintaining the pharmacological action of the natural AM. Can have significantly better pharmacokinetics, for example with respect to biological stability.

In formula (I), L is a divalent linking group. The linking group L is preferably a substituted or unsubstituted divalent hydrocarbon group, a substituted or unsubstituted divalent aliphatic hydrocarbon group, a substituted or unsubstituted divalent alicyclic group or a substituent. Alternatively, it is more preferably an unsubstituted divalent aromatic group, and further preferably a substituted or unsubstituted C ₁ to C ₂₀ alkylene group, a C ₂ to C ₂₀ alkenylene group or a C ₂ to C ₂₀ alkynylene group. preferable. The group may be one or more complex atoms (eg, O or S), a heterocycle, an amide group (-CO-NH-), an ester group (-CO-O-), or a urethane group (-O-CO). -NH-) is preferably included. When the group has a substituent, the substituent may include halogen (eg, F, Cl, Br or I), cyano, nitro and C ₁ to C ₅ alkoxy. The linking group L has an ethylene oxide unit having a repeat number in the range of 2 to 24, particularly 4 to 12, and one or more complex atoms (eg, O or S), a heterocycle, -CO-NH-,. It is preferably a divalent hydrocarbon group which may contain -CO-O- or -O-CO-NH-, and has an ethylene oxide unit having a repeat number in the range of 2 to 24, particularly 4 to 12. It is more preferable that it is a divalent hydrocarbon group which may contain one or more O or -CO-NH-. By having the linking group L having the above-mentioned characteristics, the compound represented by the formula (I) of this embodiment can be compared with, for example, biologically, while maintaining the pharmacological action of the native AM. It can have significantly better pharmacokinetics with respect to stability.

The linking group L has a carboxyl group at at least one end. The linking group L preferably has a functional group such as a carboxyl group, an amino group or a succinimide group at the other end. In particular, the linking group L preferably has a carboxyl group at one end and a succinimide group at the other end. In the case of the present embodiment, the linking group L forms an amide bond with the α-amino group at the N-terminal of the peptide moiety B via the carboxyl group, and the cysteine residue of the modifying group A via the succinimide group. By forming a thioether bond with the thiol group, it is linked to the modifying group A and the peptide moiety B.

In formula (I), the peptide moiety B has an N-terminal α-amino group, specifically, an α-amino group of an N-terminal amino acid residue forming an amide bond with a carboxyl group at the end of the linking group L. It is connected to the rest.

In formula (I), the modifying group A, which is serum albumin, is such that any functional group contained in the constituent amino acid residues of serum albumin forms a covalent bond with the functional group exemplified above at the terminal of the linking group L. Is linked to the rest by. In the modifying group A, examples of the functional group forming the linkage with the linking group L include an α-amino group at the N-terminal of serum albumin, a carboxyl group at the C-terminal, a thiol group (sulfhydryl group) at the cysteine residue, and a lysine residue. Examples thereof include the εamino group of the group, the imidazole ring of the histidine residue, the guanidino group of the arginine residue, the εcarboxyl group of the glutamate residue, or the γcarboxyl group of the aspartic acid residue.

The linking group L is a substituted or unsubstituted divalent hydrocarbon group having a carboxyl group at at least one end and a functional group exemplified above at the other end (the group is one or more complex atoms (the group is one or more complex atoms). For example, it is preferably O or S), a heterocycle, -CO-NH-, -CO-O- or -O-CO-NH-), with a carboxyl group at at least one end and the other. Substituent or unsubstituted divalent aliphatic hydrocarbon group having the functional group exemplified above, substituted or unsubstituted divalent alicyclic group or substituted or unsubstituted divalent aromatic group at the end of The group may be one or more complex atoms (eg, O or S), a heterocycle, -CO-NH-, -CO-O- or -O-CO-NH-). More preferably, substituted or unsubstituted C ₁ to C ₂₀ alkylene groups, C ₂ to C ₂₀ alkenylene groups or C ₂ to C ₂₀ having a carboxyl group at at least one end and the functional group exemplified above at the other end. Alquinylene group (the group may include one or more complex atoms (eg, O or S), a heterocycle, -CO-NH-, -CO-O- or -O-CO-NH-). It is more preferable to have. In particular, the linking group L has a carboxyl group at at least one end, a functional group exemplified above at the other end, and an ethylene oxide unit having a repetition number in the range of 2 to 24, particularly 4 to 12. A divalent hydrocarbon group which may contain one or more complex atoms (eg, O or S), a heterocycle, -CO-NH-, -CO-O- or -O-CO-NH-. It is preferable that the ethylene oxide unit has a carboxyl group at at least one end, the functional group exemplified above at the other end, and has a repetition number in the range of 2 to 24, particularly 4 to 12, and 1 More preferably, it is a divalent hydrocarbon group that may contain more than one O or -CO-NH-. In the case of this embodiment, the linking group L is, for example, the following formula:

(In the formula, # represents the linking position with the modifying group A, and * represents the linking position with the peptide moiety B.). By having the linking group L having the above-mentioned characteristics, the compound represented by the formula (I) of this embodiment can be compared with, for example, biologically, while maintaining the pharmacological action of the native AM. It can have significantly better pharmacokinetics with respect to stability.

A suitable compound represented by the formula (I) is
Modification group A is human serum albumin consisting of the amino acid sequence of SEQ ID NO: 14.
Peptide part B is as follows:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(B) A peptide consisting of the amino acid sequence of SEQ ID NO: 4, or a peptide consisting of the amino acid sequence of SEQ ID NO: 4 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(C) A peptide consisting of the amino acid sequence of SEQ ID NO: 6 or a peptide consisting of the amino acid sequence of SEQ ID NO: 6 in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(D) A peptide consisting of the amino acid sequence of SEQ ID NO: 8 or a peptide consisting of the amino acid sequence of SEQ ID NO: 8 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(E) A peptide consisting of the amino acid sequence of SEQ ID NO: 10 or a peptide consisting of the amino acid sequence of SEQ ID NO: 10 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(F) A peptide consisting of the amino acid sequence of SEQ ID NO: 12, or a peptide consisting of the amino acid sequence of SEQ ID NO: 12 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(G) In any of the peptides (a) to (f), the peptide in which the disulfide bond is substituted with an ethylene group;
(H) A peptide in which 1 to 15 amino acid residues are deleted, substituted or added in any of the peptides (a) to (g);
(I) A peptide in which the C-terminal is amidated in any of the peptides (a) to (h); and a glycine residue at the C-terminal in any of the peptides (j) (a) to (h). Peptides with added groups;
A peptide moiety derived from adrenomedullin or a modification thereof, which is a peptide selected from the group consisting of
The linking group L is a substituted or unsubstituted divalent hydrocarbon group having a carboxyl group at at least one end and a functional group such as a carboxyl group, an amino group or a succinimide group at the other end (the above group is 1). More than one complex atom (eg, O or S), heterocycle, -CO-NH-, -CO-O- or -O-CO-NH- may be included).
The peptide moiety B is linked to the rest by the N-terminal α-amino group forming an amide bond with the carboxyl group at the end of the linking group L.

A more suitable compound represented by the formula (I) is
Modification group A is human serum albumin consisting of the amino acid sequence of SEQ ID NO: 14.
Peptide part B is as follows:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(I) In the peptide of (a), the peptide in which the C-terminal is amidated; and in the peptide of (j) (a), the peptide in which the glycine residue is added to the C-terminal;
A peptide moiety derived from adrenomedullin or a modification thereof, which is a peptide selected from the group consisting of
The linking group L is a substituted or unsubstituted divalent hydrocarbon group having a carboxyl group at at least one end and a functional group such as a carboxyl group, an amino group or a succinimide group at the other end (the above group is 1). More than one complex atom (eg, O or S), heterocycle, -CO-NH-, -CO-O- or -O-CO-NH- may be included).
The peptide moiety B is linked to the rest by the N-terminal α-amino group forming an amide bond with the carboxyl group at the end of the linking group L.

A more suitable compound represented by the formula (I) is
Modification group A is human serum albumin consisting of the amino acid sequence of SEQ ID NO: 14.
Peptide part B is as follows:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(I) In the peptide of (a), the peptide in which the C-terminal is amidated; and in the peptide of (j) (a), the peptide in which the glycine residue is added to the C-terminal;
A peptide moiety derived from adrenomedullin or a modification thereof, which is a peptide selected from the group consisting of
The linking group L has a carboxyl group at at least one end and a functional group such as a carboxyl group, an amino group or a succinimide group at the other end, and has a repeating number in the range of 2 to 24, particularly 4 to 12. Divalent carbonization having a unit and may contain one or more complex atoms (eg, O or S), heterocycles, -CO-NH-, -CO-O- or -O-CO-NH- It is a hydrogen group,
The peptide moiety B is linked to the rest by the N-terminal α-amino group forming an amide bond with the carboxyl group at the end of the linking group L.

The compound represented by the formula (I) of this embodiment having the above-mentioned characteristics is a drug that is significantly superior in, for example, to biological stability, as compared with the natural AM, while maintaining the pharmacological action of the natural AM. Can have kinetics.

In each aspect of the present invention, the compound of this aspect includes not only the compound itself but also a salt thereof. When the compound of this embodiment is in the form of a salt, it is preferably a pharmaceutically acceptable salt. The counter ion of the salt of the compound of this embodiment is not limited, but is, for example, a cation such as a sodium ion, a potassium ion, a calcium ion, a magnesium ion, or a substituted or unsubstituted ammonium ion, or a chloride ion. , Bromide ion, iodide ion, phosphate ion, nitrate ion, sulfate ion, carbonate ion, hydrogen carbonate ion, perchlorate ion, formate ion, acetate ion, trifluoroacetate ion, propionate ion, lactate ion, maleic acid Ion, hydroxymaleate ion, methylmaleate ion, fumarate ion, adipate ion, benzoate ion, 2-acetoxybenzoate ion, p-aminobenzoate ion, nicotinate ion, silicate ion, ascorbic acid ion , Pamoate ion, succinate ion, salicylate ion, bismethylene salicylate ion, oxalate ion, tartrate ion, malate ion, citrate ion, gluconate ion, aspartate ion, stearate ion, palmitate ion, itaconic acid Ion, glycolic acid ion, glutamate ion, benzenesulfonic acid ion, cyclohexylsulfamate ion, methanesulfonic acid ion, ethanesulfonic acid ion, isethionate ion, benzenesulfonic acid ion, p-toluenesulfonic acid ion, or naphthalenesulfonic acid ion Anions such as are preferred. When the compound of this embodiment is in the form of a salt with the counterion, the adrenomedullin activity of the compound can be substantially equivalent to that of native AM.

In each aspect of the present invention, the compound of this aspect includes not only the compound itself but also a solvate of the compound or a salt thereof. When the compound of this embodiment or a salt thereof is in the form of a solvate, it is preferably a pharmaceutically acceptable solvate. The solvent that can form a solvent with the compound or a salt thereof is not limited, but is, for example, water, or methanol, ethanol, 2-propanol (isopropyl alcohol), dimethyl sulfoxide (DMSO), acetic acid, ethanol, and the like. Organic solvents such as amine, acetonitrile or ethyl acetate are preferred. When the compound of this embodiment or a salt thereof is in the form of a solvate with the above solvent, the adrenomedullin activity of the compound can be substantially equivalent to that of natural AM.

In each aspect of the present invention, the compound of this aspect includes not only the compound itself described above or below, but also a protected form thereof. As used herein, "protected form" means a form in which a protecting group is introduced into one or more functional groups (eg, side chain amino groups of lysine residues). Further, in the present specification, the "protecting group" is a group introduced into a specific functional group in order to prevent the undesired progress of the reaction, and is quantitatively removed under the specific reaction conditions, and the same. It means a group that is substantially stable under the reaction conditions other than the above, that is, the reaction is inactive. Protecting groups that can form the protected form of the compound are, but are not limited to, for example, t-butoxycarbonyl (Boc), 2-bromobenzyloxycarbonyl (BrZ), 9-fluorenylmethoxycarbonyl (Fmoc). ), P-Toluenesulfonyl (Tos), benzyl (Bzl), 4-methylbenzyl (4-MeBzl), 2-chlorobenzyloxycarbonyl (ClZ), cyclohexyl (cHex), and phenacyl (Pac); other amino groups Benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, benzhydryloxycarbonyl, 2- (p-biphenyl) Isopropyloxycarbonyl, 2- (3,5-dimethoxyphenyl) isopropyloxycarbonyl, p-phenylazobenzyloxycarbonyl, triphenylphosphonoethyloxycarbonyl, 9-fluorenylmethyloxycarbonyl, t-amyloxyoxycarbonyl, Diisopropylmethyloxycarbonyl, isopropyloxycarbonyl, ethyloxycarbonyl, allyloxycarbonyl, 2-methylsulfonylethyloxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, cyclopentyloxycarbonyl, cyclohexyloxycarbonyl, adamantyloxycarbonyl, isobol Nyloxycarbonyl, benzenesulfonyl, mesitylenceluphonyl, methoxytrimethylphenylsulfonyl, 2-nitrobenzenesulfonyl, 2-nitrobenzenesulphenyl, 4-nitrobenzenesulfonyl, and 4-nitrobenzenesulphenyl; as other protecting groups for the carboxyl group, methylester, Ethyl ester, t-butyl ester, p-methoxybenzyl ester, and p-nitrobenzyl ester; 2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-as other side-chain protecting groups of Arg 5-sulfonyl, 4-methoxy-2,3,6-trimethylbenzenesulfonyl, 2,2,5,7,8-pentamethylchroman-6-sulfonyl, and 2-methoxybenzenesulfonyl; as other protecting groups for Tyr , 2,6-dichlorobenzyl, t-butyl, and cyclohexyl; as other protecting groups for Cys, 4-methoxybenzyl, t-butyl, Trityl, acetamidemethyl, and 3-nitro-2-pyrisinsulphenyl; as other protecting groups for His, benzyloxymethyl, p-methoxybenzyloxymethyl, t-butoxymethyl, trityl, and 2,4-dinitrophenyl. In addition, as other protecting groups of Ser and Thr, t-butyl and the like can be mentioned. When the compound of this embodiment is in a protected form by the protecting group, the adrenomedullin activity of the compound may be substantially equivalent to that of native AM.

Also, in each aspect of the invention, the compound of this aspect also includes individual enantiomers and diastereomers of the compound, as well as mixtures of stereoisomers of the compound, such as racemates.

By having the above-mentioned characteristics, the compound of this embodiment is remarkable in terms of, for example, biological stability as compared with the natural AM, while maintaining substantially the same pharmacological action as the parent compound, the natural AM. Can have excellent pharmacokinetics.

<2. Pharmaceutical use of adrenomedullin derivative>
The compound of one aspect of the present invention can have significantly superior pharmacokinetics as compared with AM while maintaining a pharmacological action substantially equivalent to that of the parent compound AM. Therefore, another aspect of the present invention relates to a medicine containing the compound of one aspect of the present invention as an active ingredient.

When the compound of one aspect of the present invention is applied to pharmaceutical use, the compound may be used alone or in combination with one or more pharmaceutically acceptable ingredients. The pharmaceutical of this embodiment can be formulated into various dosage forms commonly used in the art, depending on the desired method of administration. Therefore, the pharmaceutical of this embodiment also comprises a compound of one aspect of the invention or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and one or more pharmaceutically acceptable carriers. Can also be provided in the form of a pharmaceutical composition containing. In the case of the present embodiment, the pharmaceutical composition is, in addition to the above-mentioned components, one or more pharmaceutically acceptable media (for example, a solvent such as sterile water or a solution such as physiological saline), an excipient, and the like. Binders, vehicles, solubilizers, preservatives, stabilizers, disintegrants, disintegrants, swelling agents, lubricants, surfactants, emulsifiers, oily liquids (eg vegetable oils), suspending agents, buffers, painless Additives such as agents, antioxidants, sweeteners and flavoring agents may be included.

The dosage form of the drug of this embodiment is not particularly limited and may be a preparation for use in parenteral administration, and may be transmucosal (for example, nasal, sublingual or oral cavity mucosa, etc.), transdermal, transdermal. It may be a preparation for use in administration of an anal (enema), transvaginal or the like, or a preparation for use in oral administration. In addition, the dosage form of the pharmaceutical product of this embodiment may be a unit-dose form or a plurality of dosage forms. Formulations for use in parenteral administration include, for example, injections such as sterile solutions or suspensions with water or other pharmaceutically acceptable liquids. Additives that can be mixed with the injection are, but are not limited to, isotonic containing, for example, physiological saline, glucose or other adjuvants (eg, D-sorbitol, D-mannitol or sodium chloride). Vehicles such as liquids, solubilizers such as alcohols (eg ethanol or benzyl alcohol), esters (eg benzyl benzoate), polyalcohols (eg propylene glycol or polyethylene glycol), polysorbate 80 or polyoxyethylene hydrogenated castor oil. Nonionic surfactants such as oily liquids such as sesame oil or soybean oil, buffers such as phosphate buffers or sodium acetate buffers, soothing agents such as benzalconium chloride or prokine hydrochloride, humans. Stabilizers such as serum albumin or polyethylene glycol, preservatives, antioxidants and the like can be mentioned. The prepared injection is usually filled in a suitable container (eg, vial or ampoule) and stored in a suitable environment until use.

Additives contained in the preparation for use in transmucosal administration include, for example, vehicles, emulsifiers, suspensions, antibacterial agents (eg, chlorobutanol), isotonic agents (eg, sodium chloride), pH regulators and Penetrants can be mentioned. Additives contained in the pharmaceutical product for use in transdermal administration include, for example, a vehicle, an antipruritic agent, an antifoaming agent, a palliative agent, a surfactant, an emulsifier, a thickener, a suspending agent, a buffer, and a viscosity. Examples thereof include excipients, moisturizers, antioxidants, chemical stabilizers, colorants and decolorizers. Examples of the additive contained in the preparation for use in transanal administration include a medium, an emulsifier and a solid fat base. Additives contained in the formulation for use in vaginal administration include, for example, vehicles, buffers, oily liquids, suspensions, wetting agents, surfactants, antioxidants, antibacterial agents and isotonic agents. be able to.

Examples of the preparation for use in oral administration include tablets, pills, powders, capsules, soft capsules, microcapsules, elixirs, liquids, syrups, slurrys and suspensions. .. If desired, the tablet may be formulated as a sugar-coated or soluble-coated sugar-coated tablet, gelatin-encapsulated tablet, enteric-coated tablet, orally disintegrating tablet (OD tablet) or film-coated tablet, or two. It may be formulated as a dosage form of a heavy tablet or a multi-layer tablet.

Additives that can be mixed with tablets, capsules, etc. are not limited, but are, for example, water, ethanol, propanol, simple syrup, glucose solution, carboxymethyl cellulose, cellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone. , Gelatin, corn starch, tragant gum and arabic gum; excipients such as crystalline cellulose, lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin or stearic acid; dried starch, alginic acid Disintegrants such as sodium, agar powder, laminaran powder, sodium hydrogen carbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose or polyvinylpyrrolidone; Disintegration inhibitors such as oil; swelling agents such as corn starch, gelatin or alginic acid; lubricants such as magnesium stearate; absorption enhancers such as quaternary ammonium salts or sodium lauryl sulfate; glycerin or starch. Moisturizers; Excipients such as starch, lactose, kaolin, bentonite or colloidal silicic acid; Lubricants such as purified talc, stearate (eg magnesium stearate), powder borate or polyethylene glycol; Sucrose, lactose Alternatively, a sweetener such as saccharin; and a flavoring agent such as peppermint, red mono oil or cherry can be mentioned. When the pharmaceutical product is a capsule, it may further contain a liquid carrier such as fat or oil.

The pharmaceutical product of this embodiment can also be formulated as a depot preparation. In this case, the drug of this embodiment in the dosage form of the depot preparation can be administered, for example, subcutaneously or intramuscularly, or by intramuscular injection. By applying the pharmaceutical product of this embodiment to a depot preparation, the adrenomedullin activity of the compound of one embodiment of the present invention can be continuously expressed over a long period of time.

As described above, the compound of one embodiment of the present invention is significantly superior to AM, for example, in terms of biological stability, while maintaining substantially the same pharmacological action as the parent compound AM. Has pharmacokinetics. Therefore, the pharmaceutical product of this embodiment is preferably formulated as a single-dose formulation, and more preferably formulated as a single-subcutaneous dosage form.

The drug of this embodiment can also be used in combination with one or more other drugs useful as a drug. In this case, the pharmaceutical product of this embodiment is a single compound containing the compound of one embodiment of the present invention or a pharmaceutically acceptable salt thereof, or a solvate thereof and one or more other agents. It may be provided in the form of a pharmaceutical, and the compound of one aspect of the present invention or a pharmaceutically acceptable salt thereof, or a solvate thereof and one or more other agents are separately prepared. It may be provided in the form of a pharmaceutical combination or a kit containing multiple formulations. In the case of a pharmaceutical combination or in the form of a kit, the respective formulations can be administered simultaneously or separately (eg, sequentially).

When the compound of one aspect of the present invention is applied to pharmaceutical use, the compound of one aspect of the present invention is not only the compound itself, but also a pharmaceutically acceptable salt of the compound and a pharmaceutically acceptable solvent thereof. Also includes Japanese products. The pharmaceutically acceptable salt of the compound of one aspect of the present invention and the pharmaceutically acceptable solvate thereof are not limited, but for example, the salt or solvate exemplified above is preferable. When the compound of one aspect of the present invention is in the form of the salt or solvate, the compound can be applied to a desired pharmaceutical use.

The pharmaceutical product of this embodiment containing the compound of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient has various symptoms prevented or treated by AM. Diseases and / or disorders can be prevented or treated as well. The symptoms, diseases and / or disorders include, but are not limited to, the following.

(1) Cardiovascular disease: heart failure, pulmonary hypertension, arteriosclerosis obliterans, Buerger's disease, myocardial infarction, lymphedema, Kawasaki disease, myocarditis, arrhythmia (for example, arrhythmia after catheter ablation surgery), atrial fibrillation, Aortitis, pulmonary hypertension, hypertension, organ damage due to hypertension, peripheral vascular disease, and arteriosclerosis.
(2) Kidney / water electrolyte system diseases: renal failure and nephritis.
(3) Brain and neurological diseases: cerebral infarction, dementia, cerebrovascular dementia, Alzheimer's disease, and encephalitis.
(4) Genitourinary disease: erectile dysfunction (ED).
(5) Gastrointestinal diseases: inflammatory diseases (eg, inflammatory bowel disease or Crohn's disease), ulcerative diseases (eg, ulcerative colitis), intestinal Behcet's disease, hepatitis, liver fibrosis, cirrhosis, and liver failure.
(6) Orthopedic disease: arthritis.
(7) Endocrine and metabolic disorders: diabetes and diabetic organ disorders (eg, diabetic nephropathy or diabetic retinopathy), and primary aldosteronism.
(8) Respiratory disorders: bronchial asthma, emphysema, pulmonary fibrosis, pneumonia, acute bronchitis, chronic bronchitis, and acute respiratory distress syndrome (ARDS).
(9) Immune disorders: Diseases associated with the complement system (eg, C3 nephropathy).
(10) Other diseases: sepsis, septic shock, autoimmune diseases, multiple organ failure, pressure ulcers, wound healing, and alopecia.

Cardiovascular diseases prevented or treated by the pharmaceuticals of this embodiment are, in particular, heart failure, myocardial infarction (eg, acute myocardial infarction), arrhythmia (eg, arrhythmia after catheter ablation surgery), atrial fibrillation, pulmonary hypertension or peripheral. It is a vascular disease. Brain / neurological disorders prevented or treated by the pharmaceuticals of this embodiment are, in particular, cerebral infarction or dementia. The gastrointestinal diseases prevented or treated by the pharmaceuticals of this embodiment are, in particular, inflammatory diseases (eg, inflammatory bowel disease or Crohn's disease), ulcerative diseases (eg, ulcerative colitis) or intestinal Bechet's disease. The endocrine and metabolic disorders prevented or treated by the pharmaceuticals of this embodiment are, in particular, diabetes and diabetic organ disorders (eg, diabetic nephropathy or diabetic retinopathy). Respiratory disorders that are prevented or treated by the pharmaceuticals of this embodiment are, in particular, pulmonary fibrosis. Other diseases prevented or treated by the pharmaceuticals of this embodiment are, in particular, sepsis or septic shock.

The pharmaceutical of this embodiment is preferably a pharmaceutical for use in the prevention or treatment of the symptoms, diseases and / or disorders described above (for example, cardiovascular disease, brain / neurological disease or digestive system disease), and heart failure. , Acute myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Bechet's disease, diabetes, diabetic nephropathy, diabetes More preferably, it is a drug for use in the prevention or treatment of sexual retinopathy, pulmonary fibrosis, sepsis or septic shock. By using the drug of this embodiment for the prevention or treatment of the symptoms, diseases and / or disorders described above, the pharmacokinetics is significantly superior to that of AM, and the preventive or therapeutic effect is substantially equivalent to that of AM. Can be expressed.

As used herein, "prevention" means substantially preventing the occurrence (onset or manifestation) of symptoms, diseases and / or disorders. Also, as used herein, "treatment" means suppressing (eg, suppressing progression), ameliorating, repairing and / or curing symptoms, diseases and / or disorders that have occurred (onset or manifestation).

The compound of one aspect of the present invention has a structure in which AM, which is a natural bioactive peptide, and serum albumin are linked via a linking group L. Therefore, the compound of one aspect of the present invention is safe and has low toxicity. Therefore, the pharmaceutical product of this embodiment containing the compound of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient has the above-mentioned symptoms, diseases and / or disorders. It can be applied to various subjects in need of prevention or treatment. The subject is a subject or patient of a human or non-human mammal (eg, a warm-blooded animal such as a pig, dog, cow, rat, mouse, guinea pig, rabbit, chicken, sheep, cat, monkey, hamadryas baboon or chimpanzee). It is preferably present, and more preferably a human patient. By administering the drug of this embodiment to the subject, the symptoms, diseases and / or disorders in the subject can be prevented or treated.

When the medication of this embodiment is administered to a subject, especially a human patient, the exact dosage and administration will be the age, gender, sign of the subject to be prevented or treated, the disease and / or the exact condition of the disorder (eg, severity). ), And many factors such as the route of administration, the attending physician should finally determine the therapeutically effective dosage and administration. Therefore, in the pharmaceutical of this embodiment, the compound of one aspect of the present invention which is an active ingredient, a pharmaceutically acceptable salt thereof, or a solvate thereof which is pharmaceutically acceptable thereof is used as a therapeutically effective dosage and administration ( For example, the dose, the number of doses, and the route of administration) are administered to the subject. For example, when the drug of this embodiment is administered to a human patient, the dose of the compound of one aspect of the present invention used as an active ingredient, a pharmaceutically acceptable salt thereof, or a solvate thereof is pharmaceutically acceptable. Is usually in the range of 0.01 to 1000 μg / kg body weight / day, for example, in the range of 0.5 to 200 μg / kg body weight / day.

The drug of this embodiment may be administered at any number of times and by a route of administration. As described above, the compound of one embodiment of the present invention is significantly superior to AM, for example, in terms of biological stability, while maintaining substantially the same pharmacological action as the parent compound AM. Has pharmacokinetics. Therefore, the drug of this embodiment is preferably administered in a single dose. Further, the drug of this embodiment is preferably administered by a parenteral route such as intravenous administration, enema administration, subcutaneous administration, intramuscular administration or intraperitoneal administration, and more preferably subcutaneous administration. The above-mentioned symptoms, diseases and / or disorders in a subject are prevented by using the drug of this embodiment containing AM or an adrenomedullin derivative as an active ingredient in the above-mentioned dosage and administration (for example, dose, frequency of administration and route of administration). Or it can be treated.

The compound of one aspect of the invention can similarly prevent or treat the symptoms, diseases and / or disorders described above that are prevented or treated by AM. Therefore, another aspect of the present invention has been described above, comprising the compound of one aspect of the invention or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient. Concerning preventive or therapeutic agents for symptoms, diseases and / or disorders. The prophylactic or therapeutic agent of one aspect of the present invention has the same characteristics as the pharmaceutical of the present aspect described above. In addition, the prophylactic or therapeutic agent of this embodiment can be used in the same dosage and administration for the same symptoms, diseases and / or disorders as the medicine of this embodiment described above.

The compound of one aspect of the present invention can be used for the prevention or treatment of the symptom, the disease and / or the disorder in the subject having the symptom, the disease and / or the disorder described above. Therefore, another aspect of the invention is pharmaceutically acceptable in an effective amount of the compound of one aspect of the invention or pharmaceutically acceptable thereof in a subject requiring the prevention or treatment of the symptoms, diseases and / or disorders described above. A method for preventing or treating the above-mentioned symptoms, diseases and / or disorders, which comprises administering a salt or a solvate thereof which is pharmaceutically acceptable. In the method of this aspect, the compound or the like of one aspect of the present invention can be administered to a subject at the same dosage and administration as the pharmaceutical of this aspect described above. The symptoms, diseases and / or disorders are preferably cardiovascular disease, brain / neurological disease, digestive system disease, endocrine and metabolic disease, respiratory system disease or other disease, such as heart failure, acute myocardial infarction, arrhythmia, and atriosphere. Fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Bechette's disease, diabetes, diabetic nephropathy, diabetic retinopathy, pulmonary fibrosis, More preferably, it is septicemia or septic shock. An effective amount of a compound of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a solvate thereof is provided to a subject in need of prevention or treatment of the above-mentioned symptoms, diseases and / or disorders. By administration, the symptoms, diseases and / or disorders can be prevented or treated.

Another aspect of the invention is the compound of one aspect of the invention or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of the symptoms, diseases and / or disorders described above, or pharmaceuticals thereof. It is an acceptable solvate. Yet another aspect of the invention is the compound of one aspect of the invention or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof, in the manufacture of a pharmaceutical for the prevention or treatment of the symptoms, diseases and / or disorders described above. The use of those pharmaceutically acceptable solvates. Yet another aspect of the invention is the compound of one aspect of the invention or a pharmaceutically acceptable salt thereof, or pharmaceuticals thereof, for the prevention or treatment of the symptoms, diseases and / or disorders described above. The use of acceptable solvates. In the compound of this aspect or the use thereof, the compound of one aspect of the present invention and the like can be used for administration to a subject at the same dosage and administration as the pharmaceutical of this aspect described above. The symptoms, diseases and / or disorders are preferably cardiovascular disease, brain / neurological disease, digestive system disease, endocrine and metabolic disease, respiratory system disease or other disease, such as heart failure, acute myocardial infarction, arrhythmia, and atriosphere. Fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Bechette's disease, diabetes, diabetic nephropathy, diabetic retinopathy, pulmonary fibrosis, More preferably, it is septicemia or septic shock. By using the compound of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof for the prevention or treatment of the symptoms, diseases and / or disorders described above. Symptoms, diseases and / or disorders can be prevented or treated.

<3. Manufacturing method of adrenomedullin derivative>
Yet another aspect of the present invention relates to a method for producing a compound according to one aspect of the present invention. The method of this embodiment includes a connecting step. In addition, the method of this embodiment may include a precursor preparation step.

[3-1. Precursor preparation process]
The method of this embodiment prepares at least one of a precursor of peptide moiety B derived from AM or a variant thereof, a precursor of modifying group A which is serum albumin, and a precursor of divalent linking group L. , Precursor preparation steps may be included.

In the method of this embodiment, "precursor of modifying group A which is serum albumin" means serum albumin itself, or in the linking step described below, modifying group A and linking group L are fused with each other. Means those derivatives that have been appropriately modified or activated to be linked. The precursor of modifying group A is preferably serum albumin itself or a protected form thereof.

In this step, the precursor of modifying group A may be prepared by means such as a transformation system usually used in the art, or may be prepared by purchasing pre-produced serum albumin or the like. good. In either case, it is included in the embodiment of this step.

In the method of this embodiment, "precursor of peptide moiety B derived from adrenomedullin or a variant thereof" means AM or the variant thereof itself, or in the linking step described below, peptide moiety B and It means their derivatives that have been appropriately modified or activated so that the linking groups L are linked to each other by a condensation reaction. The precursor of peptide moiety B is preferably AM or its modifications themselves, or a protected form thereof.

In this step, the precursor of peptide moiety B can be formed by means commonly used in the art. When the precursor of peptide portion B is AM or a variant thereof itself, for example, solid phase or liquid phase peptide synthesis methods may be used, and human or non-human mammalian tissues capable of producing AM. Alternatively, a method of purifying a natural peptide from cells may be used. Alternatively, a transformation system such as Escherichia coli or Saccharomyces cerevisiae using DNA encoding AM in human or non-human mammals capable of producing AM (eg, SEQ ID NO: 2, 5, 7, 9, 11 or 13). You may use the method of expressing a large amount of recombinant protein in. Alternatively, a peptide prepared in advance may be purchased and used. In either case, it is included in the embodiment of this step.

In the precursor of peptide moiety B formed by the above means, by disulfide-forming the thiol groups of the two cysteine residues in the amino acid sequence, the two cysteine residues in the amino acid sequence are disulfide-bonded. Can be obtained as a precursor forming the above. Further, in the precursor of the peptide portion B formed by the above means, the disulfide bond formed between the two cysteine residues in the amino acid sequence is replaced with an ethylene group, whereby the disulfide bond becomes ethylene. A precursor substituted with a group can be obtained. The disulfide reaction and the substitution reaction with an ethylene group can be carried out under the conditions usually used in the art. The disulfide reaction and the substitution reaction with an ethylene group may be carried out in this step or may be carried out in the linking step described below. Both cases are included in the embodiments of the method of the present invention.

In the method of this embodiment, the "precursor of the divalent linking group L" is the linking group between the modifying group A and the linking group L and / or the peptide moiety B and the linking group L in the linking step described below. It means a compound in which at least one end, particularly both ends, is activated so that the spaces are linked together by a condensation reaction. As described above, the linking group L has a carboxyl group at at least one end, and the carboxyl group forms an amide bond with the N-terminal α-amino group of the peptide moiety B. Therefore, in the precursor of the linking group L, the terminal linked to the peptide portion B is a carboxyl group or an activated form thereof. Examples of the activated form of the carboxyl group include N-hydroxysuccinimide ester and p-nitrophenyl ester. Further, the linking group L has the functional group exemplified above at the other end, and the functional group forms a covalent bond with any functional group contained in the constituent amino acid residue of the modifying group A. do. Therefore, in the precursor of the linking group L, the terminal linked to the modifying group A is the functional group exemplified above or an activated form thereof. Examples of the activated form of the functional group exemplified above include N-hydroxysuccinimide ester, p-nitrophenyl ester and maleimide.

In this step, the precursor of the linking group L may be prepared by using a chemical synthesis means usually used in the art, or may be prepared by purchasing the precursor prepared in advance. .. In either case, it is included in the embodiment of this step.

In this step, a precursor containing a modifying group A and a linking group L in which a modifying group A which is serum albumin and a divalent linking group L are linked may be prepared. In the case of the present embodiment, as the precursor containing the modifying group A and the linking group L, the precursor of the modifying group A and the precursor of the divalent linking group L are prepared by the procedure described above, respectively, and then the modifying group is prepared. It may be prepared by linking the precursor of A and the precursor of the divalent linking group L, or the precursor prepared in advance may be purchased or prepared. In either case, it is included in the embodiment of this step.

In this step, a precursor containing a peptide moiety B and a linking group L in which a precursor of the peptide moiety B derived from AM or a modified product thereof and a divalent linking group L are linked may be prepared. .. In the case of the present embodiment, for the precursor containing the peptide portion B and the linking group L, the precursor of the peptide portion B and the precursor of the divalent linking group L are prepared by the procedure described above, respectively, and the precursor of the peptide portion B is prepared. It may be prepared by linking the precursor and the precursor of the divalent linking group L, or the precursor prepared in advance may be purchased or prepared. In either case, it is included in the embodiment of this step.

If at least one of the precursor of modifying group A, the precursor of linking group L and the precursor of peptide moiety B is a protected form thereof, in this step, optionally, the precursor of modifying group A, the precursor of linking group L A protective step of introducing one or more protecting groups into at least one of the precursors of the precursor and the precursor of the peptide moiety B, and / or the precursor of the modifying group A, the precursor of the linking group L and the precursor of the peptide moiety B. A deprotection step may be performed to deprotect at least one or more protecting groups of at least one of the protective forms of the body. The protection and deprotection steps can be carried out by the protection and deprotection reactions commonly used in the art. The protection step and the deprotection step may be carried out in this step, or may be carried out in the connection step described below. Both cases are included in the embodiment of the method of this embodiment.

[3-2. Connection process]
The method of this embodiment is between the precursor of modifying group A, which is serum albumin, and the precursor of divalent linking group L, and with the precursor of peptide moiety B derived from adrenomedullin or a modification thereof. Containing a linking step, linking at least one of the precursors of the linking group L to give the compound of formula (I).

In this step, the means for linking the precursor of the modifying group A and the precursor of the linking group L and the precursor of the peptide portion B and the precursor of the linking group L are not particularly limited. Based on the terminal structure, activation form, etc. of each precursor, a bond-forming reaction usually used in the art may be appropriately applied.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the technical scope of the present invention is not limited to these examples.

<Experiment I: Preparation of new adrenomedulin derivative>
[Synthesis]
We requested Peptide Institute, Inc. (Osaka, Japan) to synthesize MAL-dPEG4-CO-NH-human AM (MAL-AM). MAL-AM has a maleimide group at one end and a carboxyl group at the other end.

(In the formula, * indicates the connection position with the remaining part.)
It consists of a group (MAL-dPEG4 group) having an ethylene oxide unit having 4 repetition numbers represented by, and an amino acid sequence of SEQ ID NO: 1, with the C-terminal amidated and a cysteine residue at position 16. The mature natural human adrenomedulin (hAM (1-52)), which is a peptide in which the cysteine residue at position 21 forms a disulfide bond, is the carboxyl group of MAL-dPEG4 and the carboxyl group of hAM (1-52). It has a structure linked by forming an amide bond with the α-amino group of the N-terminal tyrosine residue.

Human serum albumin (hSA) was purchased from Merck / Sigma-Aldrich (Germany). hSA consists of the amino acid sequence of SEQ ID NO: 14. MAL-AM (1.1 mol) and hSA (1.0 mol) were mixed in 30 mM PBS (pH 7.2) and N, N-dimethylformamide and reacted at 4 ° C. for 2 days. The reaction mixture was thoroughly filtered using Amicon Ultra® (Merck, Germany) and the buffer was replaced with purified water. By the above procedure, 0.94 mg of a novel human serum albumin conjugated adrenomedullin (hSA-AM) was obtained. The maleimide group at the end of MAL-AM can undergo a Michael addition reaction with the thiol group of the cysteine residue of hSA. By this reaction, the maleimide group is converted to a succinimide group, and a thioether bond is formed between the succinimide group and the thiol group, whereby MAL-AM and hSA are linked.

[Estimation of molecular weight]
In order to estimate the molecular weight of the novel adrenomedulin derivative (hSA-AM) synthesized in the previous section, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting (WB), and gel filtration chromatography (GF) were performed. .. Since the molecular weight of MAL-AM is about 6400 and the molecular weight of hSA is about 66,000, the molecular weight of hSA-AM is estimated to be about 72,000.

[Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)]
Dithiothreitol (3.09 mg) was added to hSA-AM (0.73 μg) and hSA (0.68 μg), respectively, and each was treated with Tricine sample buffer (Bio-Rad, USA) (20 μL) to prepare a sample. .. This sample was electrophoresed using a 10-20% polyacrylamide gradient gel (e-PAGEL®, ATTO, Japan). The gel after electrophoresis was stained with Coomassie Brilliant Blue. Then, the gel was imaged.

[Western blotting (WB)]
Electrophoresis was performed in the same procedure as SDS-PAGE. From the gel after electrophoresis, the protein was transferred to a polyvinylidene difluoride (PVDF) membrane (Clear Blot Membrane-P plus®, ATTO, Japan). The PVDF membrane after protein transcription was blocked with 5% skim milk (room temperature, 60 minutes) and immersed in a primary antibody (mouse anti-adrenomedullin C-terminal sequence antibody, Shionogi Pharmaceutical Co., Ltd., Japan) (4 ° C, overnight). .. This primary antibody binds to the C-terminal portion of AM. The treated membrane was rinsed with phosphate buffered saline and Tween-20 (PBST). Subsequently, this membrane was immersed in a solution of a secondary antibody (anti-mouse IgG antibody, Immuno-Star goat anti-mouse HRP Conjugate®; # 170-5047, 1: 2000, Bio-Rad, USA). (Room temperature, 60 minutes). The treated membrane was rinsed again with PBST. Finally, the membrane was immersed in Clarity western ECL substance® (# 1705060, Bio-Rad, USA) and allowed to stand in the dark for 5 minutes. The membrane was then imaged using Omega Lum ™ G.

Figure 1 shows the results of SDS-PAGE and WB. In the figure, A shows the gel stained with Coomassie Brilliant Blue after SDS-PAGE, and B shows the membrane obtained by WB. In A and B, M represents a molecular weight marker (Precision Plus Protein Dual Xtra Standards), lane 1 represents 68 ng of hSA, and lane 2 represents 180 ng of hSA-AM. As shown in FIG. 1A, both hSA and hSA-AM showed a single band at approximately the same position below the 75 kDa marker (lanes 1 and 2). In addition, as shown in Fig. 1B, the presence of the C-terminal part of AM was suggested in hSA-AM (lane 2).

[Gel Filtration Chromatography (GF)]
hSA-AM and hSA were subjected to gel filtration chromatography using a Superdex ™ 200 Increase 10/300 GL column (GE Healthcare UK, UK) in 20 mM citrate buffer (pH 7.0) containing 100 mM NaCl. Eluted. As a molecular weight marker, Gel Filtration Calibration Kit LMW (GE Healthcare UK, UK) was used.

Figure 2 shows the results of GF. In the figure, A is a chromatogram of 30 μg hSA-AM, B is a chromatogram of 30 μg hSA, and C is a chromatogram of purified hSA-AM. In the figure, the horizontal axis is the retention time (minutes) of the reverse phase HPLC, and the vertical axis is the UV absorption intensity (mV) at a wavelength of 280 nm. As shown in FIGS. 2A and 2B, hSA-AM and hSA showed similar peak profiles. In gel filtration chromatography of hSA-AM, 3 fractions of eluate containing peaks with retention times (RT) of 15 minutes, 18 minutes and 25 minutes were separated. The fractions were analyzed by enzyme immunoassay (EIA) using mouse anti-adrenomedulin C-terminal sequence antibody and anti-mouse IgG antibody. Fractions containing a 25-minute peak were negative (data not shown). Subsequently, the fraction containing the peak at RT 18 minutes, that is, the purified hSA-AM was subjected to gel filtration chromatography under the same conditions again. As a result, the peak seen at RT 15 minutes disappeared (Fig. 2C). The

<Experiment II: Example of using a novel adrenomedulin derivative>
[Experiment II-1: Intracellular cAMP concentration increasing effect by adrenomedulin derivative]
The physiological effects of AM are known to be expressed through an increase in the concentration of intracellular cAMP (Kitamura K et al., Biochem Biophys Res Commun, April 30, 1993, Vol. 192 (2), pp. See 553-560). Therefore, an AM derivative was added to a cultured cell line (HEK293 cell line) in which the AM type 1 receptor (AM1 receptor) was stably expressed, and the amount of intracellular cAMP produced was measured. HEK293 cells were placed in a 24-well plate (Thermo Fisher Scientific Co., Ltd.) coated with fibronectin in Dalvecco-modified Eagle's medium (10% bovine fetal serum, 100 U / mL penicillin G, 100 μg / mL streptomycin, 0.25 μg /). Cultivated in mL amhotericin B, 100 μg / ml hyglomycin B and 250 μg / ml genetesin added (37 ° C, humidified, 5% CO ₂ conditions). After culturing for 3 days, 90% confluent cells stimulated to accumulate intracellular cAMP were subjected to the experiment. The medium was replaced with a Hanks equilibrium salt solution containing 20 mM 4- (2-hydroxyethyl) -1-piperazine ethanesulfonic acid (HEPES) and 0.1% bovine serum albumin in the presence of 0.5 mM isobutylmethylxanthine (IBMX). , The cells were added with a given concentration of native AM or AM derivative (hSA-AM) and incubated for 15 minutes under 37 ° C., humidified, 5% CO ₂ conditions. The reaction was stopped by adding cytolysis buffer. Then, using an enzyme immunoassay (EIA) kit for measuring cAMP (GE Healthcare UK, UK), the amount of intracellular cAMP produced in HEK293 cells in each well was measured. The measurement was performed in 4 iterations, and the mean value and standard error (SEM) were calculated.

Figure 3 shows the dose-response curve of intracellular cAMP production in HEK293 cells with respect to the added concentration of natural AM or AM derivative (hSA-AM). In the figure, the black circles (●) are the average values of the experiments to which natural AM was added, the white squares (□) are the average values of the experiments to which hSA-AM was added, and the error bars represent SEM. In the figure, the horizontal axis is the logarithm (M) of the added concentration of the natural AM or AM derivative (hSA-AM), and the vertical axis is the intracellular cAMP in HEK293 cells that stably express the AM1 receptor. Production amount (fmol / well). As shown in FIG. 3, hSA-AM increased the intracellular cAMP concentration in a dose-dependent manner, similar to that of natural AM. From the dose-response curve, the pEC50 of natural AM was calculated to be 8.660, and the pEC50 of hSA-AM was calculated to be 7.208.

[Experiment II-2: Comparison of pharmacokinetics of AM and AM derivatives]
From the viewpoint of animal welfare, animal experiments were conducted with the approval of the Animal Experiment Committee of Miyazaki University (No. 2014-507, 2019-533). A 7-week-old male Wistar rat was purchased from Charles River Laboratories Japan, Inc. (Kanagawa Prefecture, Japan). Rats were bred on normal diet in a 12-hour light and 12-hour dark cycle and in the absence of specific pathogens. To determine plasma peptide concentration, 10 nmol / kg hSA-AM or 50 nmol / kg native human AM were subcutaneously administered to 5 rats each. Peripheral blood samples were taken for a given time (natural human AM: 0, 0.25, 1, 2 and 3 hours after administration, hSA-AM: 0, 1, 3, 6, 12, 24, 48, 72, 120 and after administration. It was collected from the tail vein into a test tube (Easy tube (registered trademark), containing heparin Na, Eiken, Tochigi Prefecture, Japan) at 168 hours. Plasma was obtained by centrifuging the test tube at 2,000 xg. Plasma was collected from the test tube and transferred to a test tube supplemented with 21 μg aprotinin and 0.3 g Na ₂ -EDTA for storage. Human AM concentration in rat plasma was measured using specific fluorescence immunoassay (Toso Corporation) using antibodies with different recognition sites. The first antibody binds to a cyclic structure in which the cysteine residue at position 16 and the cysteine residue at position 21 of hAM (1-52) form a disulfide bond, and the second antibody is the C-terminal portion of AM. Combine to. By using these two antibodies, full-length AM (tAM) of 52 amino acid residues and mature AM (mAM) capable of expressing activity can be distinguished and quantitatively analyzed (Ohta H et al., One-step direct). assay for mature-type adrenomedullin with monoclonal antibodies. Clin Chem., February 1999, Vol. 45 (2), p. 244-51; Kubo K et al., Biological properties of adrenomedullin coupled with polyethylene glycol. Peptides, 2014 7 Mon, Vol. 57, p. 118-21. Doi: 10.1016 / j.peptides. 2014.05.005. Epub 2014 May 27.). For each measurement, the mean value and standard error (SEM) of 5 individuals were calculated.

Figure 4 shows the change over time in the amount of natural AM or AM derivative (hSA-AM) in rat plasma after subcutaneous administration. In the figure, A is the result of natural AM and B is the result of AM derivative (hSA-AM). In the figure, the horizontal axis is the elapsed time (A: time, B: day) after subcutaneous administration, and the vertical axis is the plasma peptide concentration after administration of natural AM or AM derivative (hSA-AM) ( pM). Plasma peptide concentrations are shown as tAM (black circles or squares) and mAMs (white circles or squares), respectively. As shown in FIGS. 4A and 4B, the residual blood duration of hSA-AM was significantly longer than that of AM. The time to reach the maximum blood concentration (Tmax) of hSA-AM was about 12 hours. Serum albumin recognizes and binds to endothelial cell gp60. As a result, serum albumin is known to have excellent migration activity on endothelial cells. From the results of this experiment, it was clarified that hSA-AM, which is a novel AM derivative, has significantly superior pharmacokinetics in terms of biological stability in a single dose as compared with the natural form.

[Experiment II-3: Effect of AM derivative on suppressing increase in blood pressure in spontaneously hypertensive rats (SHR)]
The effect of a single subcutaneous dose of an AM derivative (hSA-AM) on SHR to suppress the increase in blood pressure was investigated by the following procedure. As a treatment group, a single subcutaneous dose of 50 nmol / kg hSA-AM physiological saline solution was administered to 7-week-old SHRs fed an 8% high-salt diet. As a control group, the same amount of saline was subcutaneously administered as a single dose (n = 6 in each group). Body weight and blood pressure were measured before and 3, 6, 9 and 12 days after dosing. Blood pressure was measured with a tail cuff. Figure 5 shows the effect of a single subcutaneous dose of hSA-AM on SHR in suppressing the increase in blood pressure. In the figure, A is the time course of systolic blood pressure (sBP), and B is the time course of diastolic blood pressure (dBP). The horizontal axis is the elapsed time (days), and the vertical axis shows the difference in blood pressure (A: difference in sBP, B: difference in dBP) (mmHg) obtained by subtracting the blood pressure before administration from the blood pressure on each measurement day. .. The values in the figure are the average values of each group, and the error bars represent SEM.

As shown in FIG. 5, it was confirmed that the increase in blood pressure was suppressed in both sBP and dBP in the hSA-AM-administered treatment group as compared with the physiological saline-administered control group.

[Experiment II-4: Pharmacological effect in dextran sodium sulfate (DSS) -induced colitis model]
The inflammation-improving effect of two subcutaneous doses of the AM derivative (hSA-AM) on a mouse DSS-induced colitis model was investigated by the following procedure. As a treatment group, a 50 nmol / kg hSA-AM physiological saline solution was subcutaneously administered to the back of the mouse (first dose, 0 days), and the same amount of hSA-AM physiology was administered 7 days after the first dose. Subcutaneous administration of saline solution (second administration). As a control group, the same amount of saline was administered subcutaneously twice. From the day of administration to 11 days after administration, 1% DSS was administered with drinking water, and from 11 days after administration to the end of the experiment, tap water was administered to start the preparation of a colitis model. Before administration (day 0) and 1, 3, 5, 7, 9, 11, 13 and 15 days after administration, body weight and stool properties were observed and evaluated by the scores shown in Table 1, and the total score was the disease. The activity index (disease activity index, DAI) was used. Figure 6 shows the changes in the DAI score due to subcutaneous administration of hSA-AM to DSS-induced colitis model mice. In the figure, the horizontal axis is the elapsed time (day), and the vertical axis is the DAI score. The values in the figure are the average values of each group, and the error bars represent SEM.

As shown in FIG. 6, the DAI score was significantly reduced in the hSA-AM-administered treatment group as compared with the control group. These results suggest that administration of hSA-AM reduced inflammation of the large intestine.

The present invention is not limited to the above-described embodiment, but includes various modifications. For example, the above-described embodiment has been described in detail in order to explain the present invention in an easy-to-understand manner, and is not necessarily limited to the one including all the configurations described. In addition, it is possible to add, delete, and / or replace a part of the configuration of each embodiment with another configuration.

All publications, patents and patent applications cited in this specification shall be incorporated herein by reference as is.

Claims

Equation (I):
ALB (I)
[During the ceremony,
A is a modifying group that is serum albumin,
B is a peptide moiety derived from adrenomedulin or a variant thereof.
L is a divalent linking group having a carboxyl group at at least one end.
The peptide moiety B is linked to the rest by the N-terminal α-amino group forming an amide bond with the carboxyl group at the end of the linking group L. ]
A compound represented by, or a salt thereof, or a hydrate thereof.
The compound according to claim 1, a salt thereof, or a hydrate thereof, wherein the modifying group A is human serum albumin consisting of the amino acid sequence of SEQ ID NO: 14.
The linking group L is a substituted or unsubstituted divalent hydrocarbon group (the divalent hydrocarbon group is one or more complex atoms, a heterocycle, an amide group (-CO-NH-), an ester group (-). The compound according to claim 1 or 2, or a salt thereof, or a hydrate thereof, which may contain a CO-O-) or a urethane group (-O-CO-NH-).
The linking group L has an ethylene oxide unit having a number of repetitions in the range of 2 to 24, and one or more complex atoms, heterocycles, amide groups (-CO-NH-), ester groups (-CO-O-). ), Or a divalent hydrocarbon group which may contain a urethane group (-O-CO-NH-), the compound according to claim 3 or a salt thereof, or a hydrate thereof.
Adrenomedullin or its modifications are as follows:
(I) A peptide consisting of the amino acid sequence of adrenomedulin,
(Ii) A peptide consisting of the amino acid sequence of adrenomedullin, in which two cysteine residues in the amino acid sequence form a disulfide bond.
(Iii) In the peptide of (ii), the peptide in which the disulfide bond is substituted with an ethylene group,
(Iv) A peptide in which 1 to 15 amino acid residues are deleted, substituted or added in any of the peptides of (i) to (iii).
(V) In any of the peptides (i) to (iv), the peptide in which the C-terminal is amidated, and in any of the peptides (vi), (i) to (iv), the glycine residue at the C-terminal remains. The compound according to any one of claims 1 to 4, a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of peptides to which a group is added.
Adrenomedullin or its modifications are as follows:
(I) A peptide consisting of the amino acid sequence of adrenomedulin,
(Ii) A peptide consisting of the amino acid sequence of adrenomedullin, in which two cysteine residues in the amino acid sequence form a disulfide bond.
(V) In the peptide of (i) or (ii), the peptide whose C-terminal is amidated, and in the peptide of (vi) (i) or (ii), a glycine residue is added to the C-terminal. The compound according to claim 5, a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of peptides.
Adrenomedullin or its modifications are as follows:
(Iv') In any of the peptides (i) to (iii), a peptide in which the amino acid residues at positions 1 to 15, 1 to 10 or 1 to 5 from the N-terminal side are deleted,
(V) Select from the group consisting of the peptide in which the C-terminal is amidated in the peptide of (iv') and the peptide in which the glycine residue is added to the C-terminal in the peptide of (vi) (iv'). The compound according to claim 5, a salt thereof, or a hydrate thereof, which is a peptide to be used.
Adrenomedullin or its modifications are as follows:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(B) A peptide consisting of the amino acid sequence of SEQ ID NO: 4, or a peptide consisting of the amino acid sequence of SEQ ID NO: 4 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(C) A peptide consisting of the amino acid sequence of SEQ ID NO: 6 or a peptide consisting of the amino acid sequence of SEQ ID NO: 6 in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(D) A peptide consisting of the amino acid sequence of SEQ ID NO: 8 or a peptide consisting of the amino acid sequence of SEQ ID NO: 8 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(E) A peptide consisting of the amino acid sequence of SEQ ID NO: 10 or a peptide consisting of the amino acid sequence of SEQ ID NO: 10 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(F) A peptide consisting of the amino acid sequence of SEQ ID NO: 12, or a peptide consisting of the amino acid sequence of SEQ ID NO: 12 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(G) In any of the peptides (a) to (f), the peptide in which the disulfide bond is substituted with an ethylene group;
(H) A peptide in which 1 to 15 amino acid residues are deleted, substituted or added in any of the peptides (a) to (g);
(I) A peptide in which the C-terminal is amidated in any of the peptides (a) to (h); and a glycine residue at the C-terminal in any of the peptides (j) (a) to (h). Peptides with added groups;
The compound according to any one of claims 1 to 5, a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of.
Adrenomedullin or its modifications are as follows:
(A) A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(B) A peptide consisting of the amino acid sequence of SEQ ID NO: 4, or a peptide consisting of the amino acid sequence of SEQ ID NO: 4 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(C) A peptide consisting of the amino acid sequence of SEQ ID NO: 6 or a peptide consisting of the amino acid sequence of SEQ ID NO: 6 in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(D) A peptide consisting of the amino acid sequence of SEQ ID NO: 8 or a peptide consisting of the amino acid sequence of SEQ ID NO: 8 and in which the cysteine residue at position 16 and the cysteine residue at position 21 form a disulfide bond;
(E) A peptide consisting of the amino acid sequence of SEQ ID NO: 10 or a peptide consisting of the amino acid sequence of SEQ ID NO: 10 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(F) A peptide consisting of the amino acid sequence of SEQ ID NO: 12, or a peptide consisting of the amino acid sequence of SEQ ID NO: 12 and in which the cysteine residue at position 14 and the cysteine residue at position 19 form a disulfide bond;
(I) A peptide in which the C-terminal is amidated in any of the peptides (a) to (f); and a glycine residue at the C-terminal in any of the peptides (j) (a) to (f). Peptides with added groups;
The compound according to claim 8, a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of.
Adrenomedullin or its modifications are as follows:
(H') In any of the peptides (a) to (d), the amino acid residues at positions 1 to 15, 1 to 10 or 1 to 5 from the N-terminal side are deleted, or ( In the peptide of e) or (f), the peptide in which the amino acid residues at positions 1 to 13, 1 to 8 or 1 to 5 are deleted from the N-terminal side;
(I) In the peptide of (h'), the peptide in which the C-terminal is amidated; and in the peptide of (j) (h'), the peptide in which the glycine residue is added to the C-terminal;
The compound according to claim 8, a salt thereof, or a hydrate thereof, which is a peptide selected from the group consisting of.
Between the precursor of modifying group A and the precursor of divalent linking group L, which is serum albumin, and the precursor of peptide moiety B and the precursor of divalent linking group L derived from adrenomedullin or a modification thereof. The compound according to any one of claims 1 to 10, or a salt thereof, or a salt thereof, which comprises a linking step of linking at least one of the compounds to obtain the compound represented by the formula (I). Method for producing hydrate.
A pharmaceutical product containing the compound according to any one of claims 1 to 10, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable hydrate thereof as an active ingredient.
Heart failure, acute myocardial infarction, arrhythmia, atrial fibrillation, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Bechet's disease, diabetes, diabetic nephropathy, The pharmaceutical according to claim 12, for use in the prevention or treatment of diabetic nephropathy, pulmonary fibrosis, sepsis or septic shock.
Heart failure, acute myocardial infarction, arrhythmia, atriosphere containing the compound according to any one of claims 1 to 10, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable hydrate thereof as an active ingredient. Arrhythmia, pulmonary hypertension, peripheral vascular disease, cerebral infarction, dementia, inflammatory bowel disease, Crohn's disease, ulcerative colitis, intestinal Bechette's disease, diabetes, diabetic nephropathy, diabetic retinopathy, pulmonary fibrosis, A prophylactic or therapeutic agent for diabetes mellitus or diabetic shock.