JP2003292456A - Use of apelin - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new use of apelin. <P>SOLUTION: This adrenocorticotropic hormone secretion-adjusting agent or oxytocin secretion-adjusting agent containing apelin, its modified compound, its amide, its ester or its salt is provided. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel use of apelin, a modified form thereof, an amide thereof, an ester thereof, or a salt thereof based on the adrenocorticotropic hormone secretion-regulating action or the oxytocin secretion-regulating action.

[0002]

2. Description of the Related Art Many hormones and neurotransmitters regulate biological functions through specific receptors present on cell membranes. Many of these receptors are coupled guanine nucleotide-binding proteins.
binding protein, sometimes abbreviated as G protein below)
Signal transduction in cells through activation of Recently,
Among such G protein-coupled receptors, the so-called orphan G protein-coupled receptor APJ (human receptor protein encoded by a gene [Gene, 136, 355]
Page (1993)]) was found in bovine stomach extracts [Biochem. Biophys. Res. Com.
mun., 251, p. 472 (1998)]. This peptide was named because it is an endogenous ligand of the APJ (e ndogenous li ga n d ) and Apelin (apelin). Apelin suppresses cytokine production from mouse spleen cells [Biochemica Biophysica Actor,
1452, p. 25 (1999)], and decrease of blood pressure and increase of drinking behavior [Journal of Neurochemistry, vol. 74, p. 34 (2000)]. WO99 / 33976 discloses apelin, which is a ligand for APJ, and this apelin has a central nervous function regulating action, a circulatory function regulating action, a cardiac function regulating action, an immune function regulating action, a digestive function regulating action, It is described that it is involved in the metabolic function controlling action or the reproductive organ function controlling action (Patent Document 1).
Also, WO00 / 18793 and WO01 / 7
In 0769, modified forms of apelin are reported (Patent Documents 2 and 3).

[0003]

[Patent Document 1] WO99 / 33976

[0004]

[Patent Document 2] WO00 / 18793

[0005]

[Patent Document 3] WO01 / 70769

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel use of apelin or its modified form.

[0007]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have unexpectedly found that apelin or a modified form thereof is WO99 / 33976, WO00 / 18793. Publication and WO01 / 7
It was found that they have an adrenocorticotropic hormone secretion-regulating action and an oxytocin secretion-regulating action which are not described in Japanese Patent No. 0769. The present inventors have succeeded in completing the present invention as a result of further studies based on these findings.

That is, the present invention provides (1) apelin,
Or its amide or its ester, or
Regulation of adrenocorticotropic hormone secretion containing the salt
Agent, (2) apelin, its modified form or its amide
Or low aldo containing its ester or its salt
Steronosis, hypocortisolemia, secondary and chronic accessory
Hyporenal cortex, Addison's disease, adrenal insufficiency, pain or
Or obesity preventive / therapeutic agent, (3) apelin, and modified forms thereof
Or its amide or its ester or its salt
Oxytocin secretion regulator comprising (4) aperi
Or its modified form or its amide or its ester
Or a labor inducer containing the salt, weak labor, relaxation
Mild bleeding, uterine retrograde failure, lactation failure, post-traumatic stress
Prophylactic / therapeutic agent for syndrome or milk stasis, or placenta
Before and after delivery, cesarean section, miscarriage or artificial abortion
Adjuvant, (5) contains DNA encoding apelin
Regulation of adrenocorticotropic hormone secretion containing DNA
Agent, (6) DN containing DNA encoding apelin
Hypoaldosteronism and low cortisol containing A
, Secondary and chronic hypoadrenocorticism, Addison
Disease / adrenal insufficiency, pain / obesity preventive / therapeutic agent,
(7) DNA containing DNA encoding apelin
Oxytocin secretion regulator comprising (8) apelin
A DNA-containing DNA containing
Labor inducement agent, weak labor, flaccid hemorrhage, uterine incompetence, milk content
Prediction of lactation failure, post-traumatic stress syndrome or milk stasis
Anti-therapeutic agents, or before and after delivery of placenta, cesarean section, miscarriage
The supplement for artificial abortion, (9) Apelin,
Modified product or its amide or its ester or its
Corticotropin containing an antibody against salt
Secretory regulator, (10) apelin, its modified form or its
Antibody against amide or its ester or its salt
Containing Cushing's disease, Cushing's syndrome, adrenal gland
Corticotropin producing pituitary tumor, CRH producing tumor,
Rudosterone producing tumor, aldosteronism, primary col
Tizole resistance, congenital adrenal deficiency, Addison's disease,
Stress, depression, anorexia nervosa, gastric / duodenal ulcer
(11) Ape, a preventive / therapeutic agent for ulcer or irritable bowel syndrome
Phosphorus, its modified form, its amide, or its ester
Oxytocin comprising an antibody against glutinol or its salt
Secretion regulator, (12) apelin, its modified form or its
Against amide or its ester or its salt
Overwork labor, tonic contraction, fetal hypothesis
Death, uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prad
er-Willi syndrome preventive and therapeutic agent, (13) Apelin
Antisense to DNA containing DNA
Regulation of adrenocorticotropic hormone secretion containing DNA
Agent, (14) D containing DNA encoding apelin
Cushion containing antisense DNA against NA
Disease, Cushing's syndrome, adrenocorticotropic hormone production
Pituitary tumor, CRH producing tumor, aldosterone producing tumor
Ulcer, aldosteronism, primary cortisol resistance, previous
Natural adrenal deficiency, Addison's disease, stress, depression,
Anorexia nervosa, gastric / duodenal ulcer or irritable enteropathy
(15) Apelin coding D
Includes antisense DNA to DNA containing NA
An Oxytocin Secretion Regulator Having (16) Apelin
Antisense to DNA containing DNA encoding
Labor pain, tonic uterine contractions,
Fetal distress, uterine rupture, cervical laceration, dysmenorrhea, premature birth or
Is a prophylactic / therapeutic agent for Prader-Willi syndrome, (17) Aperi
Or its modified form or its amide or its ester
Or adrenocortical stimulation comprising an antibody against its salt
Diagnostic agent for hormone secretion abnormality, (18) Apelin, its repair
Ornament or its amide or its ester or its
Of abnormal oxytocin secretion containing an antibody against salt
Diagnostic agent, (19) Apelin, its modified form or its amyl
Antibody to the peptide or its ester or its salt
Having hypoaldosteronism, hypocortisolemia,
Secondary and chronic hypoadrenocorticism, Addison's disease, accessory
Renal insufficiency, pain, obesity, weak labor, loose bleeding, uterine reversion
Old insufficiency, lactation insufficiency, post-traumatic stress syndrome, chyle
Stasis, Cushing's disease, Cushing's syndrome, adrenocortical stimulation
Hormone-producing pituitary tumor, CRH-producing tumor, aldoster
Ron-producing tumor, aldosteronism, primary cortisol
Resistance, congenital adrenal deficiency, stress, depression, god
Anorexia nervosa, gastric / duodenal ulcer, irritable bowel syndrome,
Hypertonic pain, ankylosing uterine contraction, fetal distress, uterine rupture, cervical canal
Diagnosis of lacerations, dysmenorrhea, preterm birth or Prader-Willi syndrome
Antidote, containing DNA encoding (20) apelin
Of abnormal adrenocorticotropic hormone secretion containing DNA
Diagnostic agent containing (21) DNA encoding apelin
Of abnormal oxytocin secretion containing DNA
Agent, (22) D containing DNA encoding apelin
Hypoaldosteronism and low cortisol containing NA
, Secondary and chronic hypoadrenocorticism, hydrangea
Disease, adrenal insufficiency, pain, obesity, weak labor, relaxation
Blood, uterine retrograde failure, lactation failure, post-traumatic stress symptoms
Herd, milk stasis, Cushing's disease, Cushing's syndrome, vice
Corticotropin-producing pituitary tumor, CRH-producing tumor,
Aldosterone producing tumor, aldosteronism, primary co
Lutisol resistance, congenital adrenal deficiency, stress,
Depression, anorexia nervosa, gastric / duodenal ulcer, hypersensitivity
Bowel syndrome, hypertonic labor, ankylosing uterine contraction, fetal distress, uterus
Rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-Willi
SEQ ID NO: 1 shows (23) apelin, a diagnostic agent for syndrome
Amino acid which is identical or substantially identical to the amino acid sequence
The above-mentioned (1) to (22), which is a peptide containing an acid sequence
The agent as described, (24) apelin is SEQ ID NO: 5, SEQ ID NO:
No .: 7, SEQ ID NO: 9 or SEQ ID NO: 11
The above is a peptide containing a partial sequence of the amino acid sequence
(1) to (22), the agent (25) apelin is (a)
Amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11
A sequence having an amino acid sequence from the 6th to the 77th in the sequence
Peptide, (b) represented by SEQ ID NO: 5, 7, 9 or 11.
40th to 77th amino acids in the amino acid sequence
Peptides having an acid sequence, (c) SEQ ID NOs: 5, 7, 9
Or the 42nd to the 42nd amino acid sequence represented by 11
Peptide having the 77th amino acid sequence, (d) sequence
Of the amino acid sequence represented by the number: 5, 7, 9 or 11
Pep having the 47th to 77th amino acid sequence
Chid, (e) represented by SEQ ID NO: 5, 7, 9 or 11
61st to 77th amino acids in the amino acid sequence
A peptide having a sequence, (f) SEQ ID NOs: 5, 7, 9 or
65th to 7th of the amino acid sequence represented by 11 or 11.
Peptide having the 7th amino acid sequence or its N-terminus
Amino acid in which the amino acid at the end (Gln) was oxidized with pyroglutamine
A peptide having a no acid sequence, (g) SEQ ID NOs: 5, 7,
The first to the second amino acid sequence represented by 9 or 11
Peptide having the 25th amino acid sequence, (h) sequence
Of the amino acid sequence represented by the number: 5, 7, 9 or 11
Pepti having the 6th to 25th amino acid sequences
(I) represented by SEQ ID NO: 5, 7, 9 or 11
The 42nd to 64th amino acid sequences in the amino acid sequence
A peptide having columns, (j) SEQ ID NOs: 5, 7, 9 or
Is the 61st to the 64th of the amino acid sequence represented by 11.
The peptide having the th amino acid sequence, (k) the sequence number
No .: of the amino acid sequence represented by 5, 7, 9 or 11
Pepti having the 43rd to 77th amino acid sequences
De, (l) represented by SEQ ID NO: 5, 7, 9 or 11
The 41st to 77th amino acid sequences in the amino acid sequence
Peptides having rows, (m) SEQ ID NOs: 5, 7, 9 or
Is the 66th to 77th amino acid sequence represented by 11
The peptide having the th amino acid sequence, (n) the sequence number
No .: of the amino acid sequence represented by 5, 7, 9 or 11
Pepti having the 67th to 77th amino acid sequence
(O) represented by SEQ ID NO: 5, 7, 9 or 11
The 64th to 77th amino acid sequence in the amino acid sequence
A peptide having a sequence, (p) SEQ ID NOs: 5, 7, 9 or
Is from the 63rd position to the 77th position in the amino acid sequence represented by 11.
A peptide having the th amino acid sequence, (q) the sequence number
No .: of the amino acid sequence represented by 5, 7, 9 or 11
Pepti having the 65th to 76th amino acid sequences
Or (r) represented by SEQ ID NO: 5, 7, 9 or 11
65th to 75th amino acids in the amino acid sequence
The above-mentioned (1) to (22), which is a peptide having an acid sequence
The listed agent, (26) apelin, has SEQ ID NO: 5, SEQ ID NO:
7, the amino acid sequence represented by SEQ ID NO: 9 or SEQ ID NO: 11
Amino acid sequence from the 65th to the 77th amino acid sequence
The peptide according to the above (1) to (22)
Agent (27) Apelin is pGlu Arg Pro Arg Leu Ser His
Contains the amino acid sequence represented by Lys Gly Pro Met Pro Phe.
The agent according to the above (1) to (22), which is a peptide having
(28) Apelin is SEQ ID NO: 5, SEQ ID NO: 7, sequence
The amino acid sequence represented by SEQ ID NO: 9 or SEQ ID NO: 11
Contains the 42nd to the 77th amino acid sequence of
The agent according to (1) to (22) above, which is a peptide, (2
9) The modified form of apelin has the formula X ¹ -Arg-Pro-Arg-X ^Two -Ser-His-X ^Three -Gly-Pro-X ^Four -X ⁵ (I) [wherein X ¹ Are hydrogen atoms or the same or different
1 to 25 amino acids whose side chains may be substituted
An amino acid residue or peptide chain consisting of
^Two Indicates a neutral amino acid residue whose side chain may be substituted.
Then X ^Three Is a neutral amino acid residue that may have a side chain substituted
Groups, aromatic amino acid residues whose side chains may be substituted
Or a basic amino acid residue whose side chain may be substituted
Shows, X ^Four May have a bond or side chain substituted
A neutral or aromatic amino acid residue, X ⁵ Is the side chain
Amino acid residue which may be substituted or its C-terminal
Carboxyl group is hydroxymethyl group or formyl group
Amino acid derivative reduced to, hydroxyl group or side chain
Amino acid residue that may be substituted and side chain are substituted
A dipeptide chain consisting of optional amino acid residues
Or its C-terminal carboxyl group is a hydroxymethyl group.
Or a peptide derivative reduced to a formyl group is represented by the formula
-Arg-Pro-Arg-, -Ser-His- in
Alternatively, the side chain of each amino acid residue in -Gly-Pro- is
May be substituted, provided that X ^Two Leu, X ^Three But
Lys, X ^Four Indicates Met, and X ⁵ Is Pro
Or Pro-Phe is shown, and -Arg-Pro in the formula is shown.
-Arg- is an unsubstituted -Arg-Pro-Arg-,
-Ser-His- is an unsubstituted -Ser-His-
And -Gly-Pro- is an unsubstituted -Gly-Pro-
The above is a peptide represented by
(1) to (4), (9) to (12) and (17) to
The agent according to (19), the modified form of (30) apelin has the formula P1-Arg-Pro-Arg-Leu-Phe-P2-P3-Gly-Pro-P4-P5 (II) [wherein P1 is a hydrogen atom] Or each the same or different
1 to 25 amino acids whose side chains may be substituted
Amino acid residue or peptide chain consisting of
2 is a neutral amino acid residue whose side chain may be substituted or
Represents a basic amino acid residue whose side chain may be substituted.
P3 is a neutral amino acid residue which may have a side chain substituted.
Groups, aromatic amino acid residues whose side chains may be substituted
Or a basic amino acid residue whose side chain may be substituted
Shown, P4 may have a bond or side chain substituted
Indicates a neutral or aromatic amino acid residue, and P5 has a side chain
Amino acid residue that may be substituted or its C-terminal residue
Ruboxyl group becomes hydroxymethyl group or formyl group
The reduced amino acid derivative, hydroxyl group or side chain
Amino acid residues that may be replaced and side chains have been replaced
Dipeptide chain consisting of amino acid residues that may
The C-terminal carboxyl group is a hydroxymethyl group
Represents a peptide derivative reduced to a formyl group, where
-Arg-Pro-Arg-Leu-Phe- or
The side chain of each amino acid residue in -Gly-Pro- is not replaced.
Which may be included in the peptide]
(1) to (4), (9) to (12) and (17) to
The agent according to (19), the modified form of (31) apelin has the formula Q1-Arg-Pro-Arg-Leu-Ser-Ala-Q2-Gly-Q5-Q3-Q4 (III) [wherein Q1 is a hydrogen atom] Or each the same or different
1 to 25 amino acids whose side chains may be substituted
Amino acid residue or peptide chain consisting of
2 is a neutral amino acid residue whose side chain may be substituted,
Aromatic amino acid residue or side of which chain may be substituted
Shows a basic amino acid residue whose chain may be substituted,
Q3 is a neutral or optionally substituted bond or side chain.
Or an aromatic amino acid residue, and Q4 has a side chain substituted
Optional amino acid residue or its C-terminal carboxyl group
Syl group is reduced to hydroxymethyl group or formyl group
Amino acid derivative, hydroxyl group or side chain
Optional amino acid residue and side chain may be substituted
A dipeptide chain consisting of two or more amino acid residues bound to each other
If the C-terminal carboxyl group is a hydroxymethyl group or
Shows a peptide derivative reduced to a mil group, and Q5 is a side chain
Represents a neutral amino acid residue which may be substituted,
-Arg-Pro-Arg-Leu-Ser-Ala
The side chain of each amino acid residue in-may be substituted
Or a peptide or ester thereof represented by
Is an amide or a salt thereof, (1) to (4),
(9) to (12) and (17) to (19) described agents,
(32) The modified form of apelin has the formula R1-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-R2-Pro-R3 (IV) [wherein R1 is a hydrogen atom or the same or respectively. Different
1 to 25 amino acids whose side chains may be substituted
An amino acid residue or peptide chain consisting of
2 is Cha which may be substituted, and may be substituted
, Met or Nle which may be substituted, R
3 is optionally substituted Phe (Cl), substituted
Phe which may be present, Nal which may be substituted
(2), optionally substituted Cha or substituted
The optional Tyr is shown, and -Arg-Pro-A in the formula is shown.
rg-Leu-Ser-His-Lys-Gly-Pr
The side chain of each amino acid residue in o or -Pro- is
The peptide represented by
The above which is steal or its amide or its salt
(1) to (4), (9) to (12) and (17) to
The agent according to (19), (33) apelin, or a modified form thereof.
Or its amide or its ester or its salt and
(Or) the amino acid sequence represented by SEQ ID NO: 3
Contains the same or substantially the same amino acid sequence as
Apelin receptor, its partial peptide or its salt
A corticotropin secretion regulator characterized by
Screening method for (34) encoding apelin
DNA containing DNA and / or sequence number
No .: the same as or substantially the same as the amino acid sequence represented by 3.
Or an apelin receptor containing an amino acid sequence identical to
The feature is that the DNA encoding the partial peptide is used.
Of the adrenocorticotropic hormone secretion regulator
Method, (35) apelin, its modified form or its
Amide or its ester or its salt and (or
Or the same as the amino acid sequence represented by SEQ ID NO: 3.
Alternatively, an apery containing substantially the same amino acid sequence
Use of a receptor or its partial peptide or its salt
, A oxytocin secretion regulator characterized by
(36) containing DNA encoding apelin
And / or SEQ ID NO: 3
Amino acid that is identical or substantially identical to the amino acid sequence
Apelin receptor containing a sequence or a partial peptide thereof
Characterized by using a DNA encoding
Screening method for syn secretion regulator, (37) substance
Is apelin, its modified form or its amide or its
Represented by SEQ ID NO: 3
Amino acid sequence identical or substantially identical to the amino acid sequence
Apelin receptors containing rows, partial peptides thereof or
The above-mentioned (33) which is a substance that changes the binding property with the salt.
To (36) the screening method, (38) the substance is
Apelin or its salt and / or SEQ ID NO:
The same or substantially the same as the amino acid sequence represented by 3.
An apelin receptor containing a single amino acid sequence or the
The above (33) to (36), which are substances that regulate the expression of salts
The screening method described in (39) apelin,
Modified product or its amide or its ester or its
And / or the amino acid represented by SEQ ID NO: 3.
Amino acid sequence identical or substantially identical to the amino acid sequence
Apelin receptor containing, its partial peptide or its
Adrenocorticotropic hormone content characterized by containing salt
Kit for screening for secretion regulator, (40)
DNA containing DNA encoding phosphorus and (and also
Is the same as the amino acid sequence represented by SEQ ID NO: 3.
Preferably containing substantially the same amino acid sequence
It contains DNA encoding the receptor or its partial peptide.
Adrenocortical stimulation characterized by containing a DNA having
A screening kit for hormone secretion regulators, (4
1) Apelin, its modified form, its amide, or
The ester or salt thereof and / or the sequence number
No .: the same as or substantially the same as the amino acid sequence represented by 3.
An apelin receptor containing an amino acid sequence identical to
Characterized by containing a partial peptide or a salt thereof
A kit for screening an oxytocin secretion regulator,
(42) DN containing DNA encoding apelin
A and / or amino represented by SEQ ID NO: 3
Contains an amino acid sequence that is identical or substantially identical to the acid sequence.
Having an apelin receptor or a partial peptide thereof
Characterized by containing a DNA containing
For screening oxytocin secretion regulators
(43) The screen according to (33) or (34) above.
Or the method described in (39) or (40) above
Adrenal cortex stimulation obtained using a screening kit
Hormone secretion regulator (44) Above (33) or (3
4) the screening method described above or (39) or
Was obtained using the screening kit described in (40).
An oxytocin secretion regulator, (45) above (43)
Corticotropin secretion containing the listed substances
Containing agent, (46) comprising the substance described in (43) above
Hypoaldosteronism, hypocortisolemia, secondary and
And chronic hypoadrenal cortex, Addison's disease, and adrenal insufficiency
Preventive / therapeutic agent for all, pain or obesity, (47) above (4
3) Cushing's disease, Koussin, containing the substances described
Guz syndrome, adrenocorticotropic hormone-producing pituitary tumor, CR
H-producing tumor, aldosterone-producing tumor, aldosterone
Disease, primary cortisol resistance, congenital adrenal enzyme deficiency
, Addison's disease, stress, depression, anorexia nervosa
, Cure of gastric / duodenal ulcer or irritable bowel syndrome
Medical agent, (48) containing the substance described in (44) above
Oxytocin secretion regulator, (49) As described in (44) above.
Labor inducer containing substances, weak labor, relaxation bleeding,
Uterine failure, lactation failure, post-traumatic stress syndrome
Or preventative or therapeutic agent for milk stasis, or placental prepartum
Later, Caesarean section, assistance for miscarriage or abortion
Agent, (50) an agent containing the substance described in (44) above.
Forced labor, ankylosing uterine contraction, fetal distress, uterine rupture, cervical cleft
Prevention of wounds, dysmenorrhea, preterm birth or Prader-Willi syndrome
-Therapeutic agents, (51) (1) apelin, modified forms thereof, or
Its amide or its ester or its salt, and a sequence
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
That alters the binding properties with partial peptides or salts thereof
Quality, or (2) apelin or its salt and (or
Is the same as the amino acid sequence represented by SEQ ID NO: 3.
Preferably containing substantially the same amino acid sequence
Containing a substance that regulates the expression of the receptor or its salt
Adrenocorticotropin secretion regulator, (52) (1)
Apelin, its modified form or its amide or its
Stell or its salt and the amino acid represented by SEQ ID NO: 3
Amino acid sequence identical or substantially identical to the amino acid sequence
Apelin receptor containing, its partial peptide or its
Substances that change the binding properties with salt, or (2) aperi
Represented by SEQ ID NO: 3 or a salt thereof and / or SEQ ID NO: 3.
Amino acid sequence identical or substantially identical to the
Expression of apelin receptor or its salt containing no acid sequence
Oxytocin secretion control comprising a substance that regulates
And (53) the amino acid sequence represented by SEQ ID NO: 3
An array containing identical or substantially identical amino acid sequences
An agonist for the perine receptor or its salt, or
(2) Apelin or its salt and / or sequence
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
Contains a substance having an activity of promoting the expression of the salt,
Hypoaldosteronism, hypocortisolemia, secondary
And chronic hypoadrenocorticism, Addison's disease, adrenal gland
Preventive / therapeutic agent for incompetence, pain or obesity, (54) sequence
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
Is an agonist for the salt, or (2) apelin
Or a salt thereof and / or represented by SEQ ID NO: 3
Amino acid identical or substantially identical to the amino acid sequence
Expression of apelin receptors containing acid sequences or salts thereof
Induction of labor, which contains a substance having a promoting activity
Agent, weak labor, laxative bleeding, uterine incompetence, non-lactation
Prevention of post-traumatic stress syndrome or milk stasis
Before or after delivery of placenta, Caesarean section, miscarriage
Is an adjuvant for artificial abortion, (55) SEQ ID NO: 3
Identical or substantially identical amino acid sequence
Pairs with apelin receptors containing mino acid sequences or salts thereof
Antagonist, or (2) apelin or its
And / or the amino acid represented by SEQ ID NO: 3.
Amino acid sequence identical or substantially identical to the amino acid sequence
Inhibits expression of contained apelin receptor or its salt
Cushing's disease, Kush, which contains an active substance
Sing syndrome, adrenocorticotropic hormone-producing pituitary tumor,
CRH producing tumor, aldosterone producing tumor, aldostere
Rhonosis, primary cortisol resistance, congenital adrenal enzyme deficiency
Deficiency, Addison's disease, stress, depression, anorexia nervosa
, Cure of gastric / duodenal ulcer or irritable bowel syndrome
Medical agent, (56) Amino acid sequence represented by SEQ ID NO: 3
Contains the same or substantially the same amino acid sequence as
An antagonist for the apelin receptor or a salt thereof,
Or (2) apelin or its salt and / or
It may be the same as the amino acid sequence represented by SEQ ID NO: 3.
Is an apelin receptor containing a substantially identical amino acid sequence
A substance having an activity of inhibiting the expression of the body or a salt thereof,
Containing excessive labor pain, ankylosing uterine contraction, fetal distress, child
Miya rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-Wil
Preventive / therapeutic agent for li syndrome, (57) for mammals,
Apelin, its modified form or its amide or its
Characterized by administering an effective amount of steal or its salt
A method for regulating adrenocorticotropic hormone secretion, (58) feeding
In addition, apelin, its modified form or its amide
Or an effective amount of its ester or its salt
Hypoaldosteronism, hypocortisolemia characterized by
, Secondary and chronic hypoadrenocorticism, Addison
Disease, adrenal insufficiency, pain / obesity prevention / treatment method,
(59) For mammals, apelin, its modified form, or
Is effective of its amide or its ester or its salt
Of oxytocin secretion characterized by administering a large amount
Method (60) For mammals, apelin, and modified forms thereof
Or its amide or its ester or its salt
Labor induction method characterized by administering effective amount, weak
Labor, laxative bleeding, uterine restoration failure, lactation failure, post-traumatic injury
How to prevent and treat stress syndrome or milk stasis,
Or before and after delivery of placenta, cesarean section, miscarriage or artificial pregnancy
Assistance methods for abortion, (61)
DNA containing DNA encoding perine or
Same as or identical to the amino acid sequence represented by SEQ ID NO: 3
Apelin receptors containing qualitatively identical amino acid sequences
Or D containing DNA encoding the partial peptide
Adrenal cortex stimulation characterized by administering an effective amount of NA
Hormone secretion control method (62) For mammals,
DNA containing DNA encoding apelin or
The same as the amino acid sequence represented by SEQ ID NO: 3 or
Apelin receptors containing substantially identical amino acid sequences
Or containing DNA encoding the partial peptide
Low aldos characterized by administering an effective amount of DNA
Teronosis, hypocortisolemia, secondary and chronic adrenal glands
Hypocortical, Addison's disease, adrenal insufficiency, pain or
Is a method for preventing and treating obesity, (63) for mammals,
DNA containing DNA encoding apelin or
It may be the same as the amino acid sequence represented by SEQ ID NO: 3.
Is an apelin receptor containing a substantially identical amino acid sequence
Contains DNA encoding the body or its partial peptides
Oxyto, characterized in that it administers an effective amount of DNA
Synthetic secretion control method, (64)
DNA or sequence containing DNA encoding phosphorus
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
Is a DN containing DNA encoding the partial peptide
A method for inducing labor, which comprises administering an effective amount of A.
Weak labor, flaccid hemorrhage, uterine retrograde failure, lactation failure, external
How to prevent and treat post-injury stress syndrome or milk retention
Method or placental delivery, cesarean section, miscarriage or person
Auxiliary methods for abortion, (65) For mammals,
Characterized by administering an effective amount of the substance according to (43) above
A method for regulating adrenocorticotropic hormone secretion, (66)
An effective amount of the substance described in (43) above is administered to dairy animals.
Hypoaldosteronism, low corti
Zoremia, secondary and chronic hypoadrenocorticism,
Prevention and treatment of Dison disease, adrenal insufficiency, pain or obesity
The method according to (43) above, wherein the method is (67) for mammals
Cushing characterized by administering an effective amount of a substance
Disease, Cushing's syndrome, adrenocorticotropic hormone ptosis
Body tumor, CRH producing tumor, aldosterone producing tumor,
Rudosteronism, primary cortisol resistance, congenital accessory
Renal enzyme deficiency, Addison's disease, stress, depression, neurogenic
Anorexia, gastroduodenal ulcer or irritable bowel syndrome
Preventive / therapeutic methods, (68) For mammals, the above (4
4) The administration of an effective amount of the described substance
Method for regulating xytocin secretion (69) For mammals,
Characterized by administering an effective amount of the substance according to (44) above
Labor induction method, weak labor, flaccid hemorrhage, uterine retrograde
Total, lactation deficiency, posttraumatic stress syndrome or milk
Prevention and treatment of stasis, or placenta before and after delivery, Caesarean section
Assistance for open surgery, miscarriage or abortion, (7
0) Effectiveness of the substance described in (44) above for mammals
Labor pain, ankylosing uterus, characterized by high dose
Shrinkage, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth
Or the prevention and treatment of Prader-Willi syndrome, (71)
For mammals, (1) apelin, its modified form or
Its amide or its ester or its salt, and a sequence
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
That alters the binding properties with partial peptides or salts thereof
Quality, or (2) apelin or its salt and (or
Is the same as the amino acid sequence represented by SEQ ID NO: 3.
Preferably containing substantially the same amino acid sequence
An effective amount of a substance that regulates the expression of the receptor or its salt is administered.
Regulation of adrenocorticotropic hormone secretion
Method, (72) For mammals, (1) Apelin,
Or its amide or its ester, or
The salt is the same as the amino acid sequence represented by SEQ ID NO: 3.
An amino acid sequence containing one or substantially the same amino acid sequence
Binding to phosphorus receptor, its partial peptide or its salt
Substance that alters, or (2) apelin or its
Salt and / or amino acid represented by SEQ ID NO: 3
Contains an amino acid sequence that is identical or substantially identical to the acid sequence.
Which regulates the expression of the apelin receptor or its salt
Oxytocin content characterized by administering an effective amount of quality
(73) For mammals, SEQ ID NO: 3
Identical or substantially identical to the amino acid sequence represented by
Aperin receptor containing the amino acid sequence of
Agonist for peptide or its salt, or (2)
Apelin or its salt and / or SEQ ID NO:
The same or substantially the same as the amino acid sequence represented by 3.
An apelin receptor containing a single amino acid sequence or the
Administer an effective amount of a substance having an activity of promoting salt expression
Hypoaldosteronism and low cortisol characterized by
, Secondary and chronic hypoadrenocorticism, hydrangea
Disease prevention, adrenal insufficiency, pain or obesity
Method (74) for mammals, (1) with SEQ ID NO: 3
Identical or substantially identical to the represented amino acid sequence
Apelin receptor containing an amino acid sequence, partial pep
Agonist for tide or its salt, or (2)
Apelin or its salt and / or SEQ ID NO: 3
Identical or substantially identical to the amino acid sequence represented by
Receptor containing the amino acid sequence of
An effective amount of a substance having the activity of promoting the expression of
Labor induction method, weak labor, flaccid bleeding,
Uterine failure, lactation failure, post-traumatic stress syndrome
How to prevent / treat milk stasis, or placenta before delivery
After that, Caesarean section, assistance for miscarriage or artificial abortion
Method (75) for mammals, (1) with SEQ ID NO: 3
Identical or substantially identical to the represented amino acid sequence
Apelin receptor containing an amino acid sequence, partial pep
An antagonist to tid or a salt thereof, or
(2) Apelin or its salt and / or sequence
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
Is an effective amount of a substance having an activity of inhibiting the expression of the salt.
Cushing's disease characterized by administration, Cushing's disease
Group, adrenocorticotropic hormone-producing pituitary tumor, CRH production
Live tumor, aldosterone-producing tumor, aldosteronism,
Primary cortisol resistance, congenital adrenal deficiency,
Dison disease, stress, depression, anorexia nervosa, stomach /
How to prevent and treat duodenal ulcer or irritable bowel syndrome,
(76) for mammals, (1) represented by SEQ ID NO: 3
Amino acid sequence identical or substantially identical to the
Apelin receptor containing a no acid sequence, its partial peptide
Or an antagonist to a salt thereof, or (2)
Apelin or its salt and / or SEQ ID NO: 3
Identical or substantially identical to the amino acid sequence represented by
Receptor containing the amino acid sequence of
An effective amount of a substance having the activity of inhibiting the expression of
Labor pain, ankylosing uterine contraction, fetal hypothesis
Death, uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prad
Prevention and treatment of er-Willi syndrome, (77) Adrenal cortex puncture
Apelin for producing a hyperhormone secretion regulator, which
Modified product or its amide or its ester or its
Salt use, (78) hypoaldosteronism, low cortisol
, Secondary and chronic hypoadrenocorticism, horse mackerel
Prophylactic / therapeutic agent for Son's disease, adrenal insufficiency, pain or obesity
For producing apelin, its modified form or its amyl
Use of a salt or its ester or its salt, (79)
Apelin for the production of oxytocin secretion regulator,
Or its amide or its ester, or
Use of the salt, (80) labor inducer, weak labor, relaxation
Blood, uterine retrograde failure, lactation failure, post-traumatic stress symptoms
Group / milk stasis preventive / therapeutic agent or placental delivery
Assistance before and after, Caesarean section, miscarriage or artificial abortion
Apelin, its modified form or its
Use of amide or its ester or its salt, (8
1) For producing a corticotropin secretion regulator
DNA containing DNA encoding apelin or
It may be the same as the amino acid sequence represented by SEQ ID NO: 3.
Is an apelin receptor containing a substantially identical amino acid sequence
Contains DNA encoding the body or its partial peptides
Use of DNA, (82) hypoaldosteronism, low col
Thisoremia, secondary and chronic hypoadrenocorticism,
Prevention and treatment of Addison's disease, adrenal insufficiency, pain or obesity
DNA encoding apelin for the manufacture of therapeutic agents
DNA contained or amino acid represented by SEQ ID NO: 3
Amino acid sequence identical or substantially identical to the amino acid sequence
The contained apelin receptor or its partial peptide
Use of DNA containing DNA to be charged, (83) oxy
Encodes apelin for producing tocin secretion regulator
DNA containing DNA or represented by SEQ ID NO: 3
Identical or substantially identical amino acid sequence
An apelin receptor containing a mino acid sequence or a partial peptide thereof
Use of DNA containing DNA encoding a peptide,
(84) Labor inducement agent, weak labor, loose bleeding, uterine retrograde
Total, lactation deficiency, posttraumatic stress syndrome or milk
Preventive or therapeutic agent for stasis, or placenta before and after delivery, cesarean section
Manufacture adjuvants for surgery, miscarriage or abortion
DNA containing the DNA encoding the apelin
Or the same as the amino acid sequence represented by SEQ ID NO: 3
Preferably containing substantially the same amino acid sequence
It contains DNA encoding the receptor or its partial peptide.
Use of DNA carried by (85) adrenocorticotropic hormone
Use of the substance according to (43) above for producing a secretion regulator.
(86) Hypoaldosteronism, Hypocortisol blood
, Secondary and chronic hypoadrenocorticism, Addison
Manufacture of prophylactic / therapeutic agents for disease, adrenal insufficiency, pain or obesity
Using the substance according to (43) above for manufacturing, (87)
Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone
Pituitary tumor, CRH-producing tumor, aldosterone production
Live tumor, aldosteronism, primary cortisol resistance
, Congenital adrenal deficiency, Addison's disease, stress, caries
Depression, anorexia nervosa, gastric / duodenal ulcer or irritation
The above (4) for producing a prophylactic / therapeutic agent for genital bowel syndrome
3) Use of the described substance, (88) Regulation of oxytocin secretion
The use of the substance according to the above (44) for producing an agent,
(89) Labor inducement agent, weak labor, relaxation bleeding, uterine retrograde
Total, lactation deficiency, posttraumatic stress syndrome or milk
Preventive or therapeutic agent for stasis, or placenta before and after delivery, cesarean section
Manufacture adjuvants for surgery, miscarriage or abortion
(44) Use of the substance described in (44) above,
Pain, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration,
Prevention and cure of dysmenorrhea, preterm birth or Prader-Willi syndrome
Use of the substance according to (44) above for producing a therapeutic agent,
(91) Production of corticotropin secretion regulator
(1) Apelin, its modified form or its amide
Represented by SEQ ID NO: 3.
Amino acid sequence identical or substantially identical to the
Apelin receptor containing a no acid sequence, its partial peptide
Or a substance that changes the binding property with its salt, or
(2) Apelin or its salt and / or sequence
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
Use of substances that regulate the expression of its salts, (92) oxy
(1) Apelin for producing a tocin secretion regulator,
Or its amide or its ester, or
The salt is the same as the amino acid sequence represented by SEQ ID NO: 3.
An amino acid sequence containing one or substantially the same amino acid sequence
Binding to phosphorus receptor, its partial peptide or its salt
Substance that alters, or (2) apelin or its
Salt and / or amino acid represented by SEQ ID NO: 3
Contains an amino acid sequence that is identical or substantially identical to the acid sequence.
Which regulates the expression of the apelin receptor or its salt
Quality use, (93) hypoaldosteronism, low cortisol
, Secondary and chronic hypoadrenocorticism, hydrangea
Disease, adrenal insufficiency, pain or obesity
(1) Amino acid represented by SEQ ID NO: 3 for production
Contains an amino acid sequence that is identical or substantially identical to the acid sequence.
Having an apelin receptor, its partial peptide or its salt
Agonist, or (2) apelin or its
And / or the amino acid represented by SEQ ID NO: 3.
Amino acid sequence identical or substantially identical to the amino acid sequence
Promotes expression of contained apelin receptor or its salt
Use of active substances, (94) labor inducer, weak
Pain, laxative bleeding, uterine restoration failure, lactation failure, post-traumatic scan
Prophylactic / therapeutic agent for Torres syndrome or milk retention,
Before and after delivery of placenta, cesarean section, miscarriage or artificial pregnancy
Table 1 with (1) SEQ ID NO: 3 for producing a time-dependent adjunct
Identical or substantially identical amino acid sequence
Apelin receptor containing a mino acid sequence, its partial pepti
Or an agonist for a salt thereof, or (2)
Perine or a salt thereof and / or SEQ ID NO: 3
Identical or substantially identical to the represented amino acid sequence
Of an apelin receptor containing an amino acid sequence or a salt thereof
Use of a substance having an activity of promoting expression, (95)
Sing's disease, Cushing's syndrome, adrenocorticotropic hormone production
Live pituitary tumor, CRH producing tumor, aldosterone producing tumor
Ulcer, aldosteronism, primary cortisol resistance, previous
Natural adrenal deficiency, Addison's disease, stress, depression,
Anorexia nervosa, gastric / duodenal ulcer or irritable enteropathy
(1) SEQ ID NO: for producing a prophylactic / therapeutic agent for symptoms
The same or substantially the same as the amino acid sequence represented by 3.
Apelin receptor containing a single amino acid sequence, part thereof
An antagonist to the peptide or its salt, or
(2) Apelin or its salt and / or sequence
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
Is the use of a substance having an activity of inhibiting the expression of the salt,
(96) Excessive labor pain, ankylosing uterine contraction, fetal distress, uterine rupture
Cleft, cervical laceration, dysmenorrhea, preterm birth or Prader-Willi disease
(1) SEQ ID NO: for producing a prophylactic / therapeutic agent for symptoms
The same or substantially the same as the amino acid sequence represented by 3.
Apelin receptor containing a single amino acid sequence, part thereof
An antagonist to the peptide or its salt, or
(2) Apelin or its salt and / or sequence
The same as or substantially the same as the amino acid sequence represented by No. 3:
Apelin receptors containing amino acid sequences that are
Is the use of a substance having an activity of inhibiting the expression of the salt,
(97) Modulating mammalian apelin receptor activity
A method for regulating adrenocorticotropic hormone secretion, characterized by
(98) Regulation of activity of mammalian apelin receptor
A method for regulating oxytocin secretion, characterized by
Characterized by activating the apelin receptor in mammals
Hypoaldosteronism, hypocortisolemia, secondary and
And chronic hypoadrenal cortex, Addison's disease, and adrenal insufficiency
Prevention, treatment of all, pain or obesity, (100) feeding
A component characterized by activating animal apelin receptors
Labor induction method, weak labor, flaccid hemorrhage, uterine retrograde failure, milk
Insufficiency, post-traumatic stress syndrome or lactation
Prophylactic / therapeutic methods, or placenta delivery, cesarean section, flow
Assistance methods for childbirth or artificial abortion, (101) Feeding
A cookie that is characterized by inhibiting the apelin receptor of animals.
Sing's disease, Cushing's syndrome, adrenocorticotropic hormone production
Live pituitary tumor, CRH producing tumor, aldosterone producing tumor
Ulcer, aldosteronism, primary cortisol resistance, previous
Natural adrenal deficiency, Addison's disease, stress, depression,
Anorexia nervosa, gastric / duodenal ulcer or irritable enteropathy
Method of preventing / treating symptom groups, and (102)
Overwork labor, characterized by inhibiting perine receptors
Direct uterine contraction, fetal distress, uterine rupture, cervical laceration, menstrual trouble
How to prevent and treat intractable disease, preterm birth or Prader-Willi syndrome
I will provide a.

Further, the present invention provides (103) X¹Is the side chain
Is an amino acid residue that may be substituted (2
9) Described agent, (104) X¹May be replaced
pGlu or Gln in which the side chain may be substituted
The agent according to (29) above, (105) X¹Is the formula Y¹−
Y^Two(In the formula, Y¹Have the same or different side chains
Consists of 1 to 17 optionally substituted amino acids
Amino acid residue or peptide chain^TwoIs that
The same or different side chains may be substituted 1.
Amino acid residue or pepti consisting of 8 amino acids
(Showing a chain), the agent according to the above (29),
6) Y^TwoIs the formula B¹-B^Two-B^Three-B^Four-B⁵-B
⁶-B⁷-B⁸(In the formula, B¹Through B⁸Are the same
Or, an amino acid residue in which the side chain may be substituted differently
Group B), formula B^Two-B^Three-B^Four-B⁵-B⁶-B
⁷-B⁸(In the formula, each symbol has the same meaning as above),
Formula B^Three-B^Four-B⁵-B⁶-B⁷-B⁸(In the formula, each note
No. has the same meaning as above), Formula B^Four-B⁵-B⁶−
B⁷-B⁸(In the formula, each symbol has the same meaning as above),
 Formula B⁵-B⁶-B⁷-B⁸(In the formula, each symbol is the same as above.
Show significance), Formula B⁶-B⁷-B⁸(Each symbol in the formula
Has the same meaning as above), Formula B⁷-B⁸(Each in the formula
Symbol has the same meaning as above) or formula B⁸(In the formula,
B⁸Is a peptide chain represented by the same meaning as above)
The agent described in (105) above, (107) B¹Is the side chain
The above-mentioned (1) which is a neutral amino acid residue which may be replaced.
06) described agent, (108) B¹May be replaced
(109) B, which is Gly.
^Two, B^ThreeAnd B^FourAre the same or different side chains
Is a basic amino acid residue which may be substituted
(106) Description agent, (110) B⁵Is an aromatic side chain
The agent according to (106) above, which is an amino acid residue having
(111) B⁶And B⁷Are the same or different
Is a basic amino acid residue which may have a side chain substituted.
(112) B as described in (106) above.⁸Is the side chain
The above-mentioned (106), which is Gln which may be replaced.
Agent, (113) Y¹Is the expression (a) A¹-A^Two-A^Three-A^Four
-A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰-A¹¹-A
¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷(formula
Medium, A¹Through A¹⁷Are on the same or different sides
The amino acid residue of which the chain may be substituted is shown), (b)
Formula A^Two-A^Three-A^Four-A⁵-A⁶-A⁷-A⁸-A⁹
-A¹⁰-A¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵−
A¹⁶-A¹⁷(In the formula, each symbol has the same meaning as above.
), (C) Formula A^Three-A^Four-A⁵-A⁶-A⁷-A⁸−
A⁹-A¹⁰-A¹¹-A¹²-A^Thirteen-A¹⁴-A
¹⁵-A¹⁶-A ¹⁷(In the formula, each symbol has the same meaning as above.
Shown), (d) Formula A^Four-A⁵-A⁶-A⁷-A⁸-A⁹
-A¹⁰-A¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵−
A¹⁶-A ¹⁷(In the formula, each symbol has the same meaning as above.
), (E) Formula A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰
-A¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶−
A¹⁷(In the formula, each symbol has the same meaning as described above), Formula (f)
 A⁶-A⁷-A⁸-A⁹-A^{1 0}-A¹¹-A¹²−
A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each note
No. has the same meaning as above), (g) Formula A⁷-A⁸-A⁹
-A¹⁰-A¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵−
A¹⁶-A¹⁷(In the formula, each symbol has the same meaning as above.
S), (h) formula A⁸-A⁹-A¹⁰-A¹¹-A¹²−
A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each note
No. has the same meaning as above), (i) Formula A⁹-A^{1 0}-A
¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as above), (j) formula
A¹⁰-A¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵-A
¹⁶-A¹⁷(In the formula, each symbol has the same meaning as described above),
(k) Formula A^{1 1}-A¹²-A^Thirteen-A¹⁴-A¹⁵-A
¹⁶-A¹⁷(In the formula, each symbol has the same meaning as described above),
(l) Formula A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as described above), (m) formula
A^Thirteen-A¹⁴-A¹⁵-A^{1 6}-A¹⁷(In the formula, each note
No. has the same meaning as above), (n) formula A¹⁴-A¹⁵−
A¹⁶-A¹⁷(In the formula, each symbol has the same meaning as above.
S), (o) formula A¹⁵-A^{1 6}-A¹⁷(Each symbol in the formula
Has the same meaning as above), (p) Formula A¹⁶-A¹⁷(formula
Where each symbol has the same meaning as above) or (q) A
¹⁷(A¹⁷Represents the same meaning as above)
The acid residue or peptide chain according to (105) above.
Agent, (114) A¹Is an amino acid residue having an aromatic side chain
The agent according to (113) above, which is a base, (115) A^TwoAnd
And A^ThreeIs a neutral amino acid residue whose side chain may be substituted
The agent according to (113) above, which is^FourIs the side chain
Neutral or basic -L-amino which may be substituted
The agent according to (113) above, which is an acid residue, (117) A.⁵
Is Pro in which the side chain may be substituted (11
3) Described agent, (118) A⁶And A⁹Are the same
One or different, basic with optionally substituted side chains
The agent according to (113) above, which is an amino acid residue, (11
9) A⁷And A¹⁰Are the same or different,
Amino acid residue or side chain having a hydroxy group in the side chain
The above (1) which is a neutral amino acid residue which may be substituted.
13) Described agent, (120) A⁸Has a hydroxy group on the side chain
The agent according to (113) above, which is an amino acid residue having
(121) A⁸Is Ser, Pro or Hyp
The agent described in (113), (122) A.¹⁰Is hydroxy
The above (113) which is an amino acid residue having a group in the side chain
Listed agent, (123) A¹¹Through A¹⁴Are the same
Or a neutral amino optionally substituted on the side chain
The agent according to (113) above, which is an acid residue, (124) A.
¹⁵Is an amino acid residue having an aromatic side chain
(113) Agent, (125) A¹⁵Is Trp
The agent described in (113) above, (126) A.¹⁶And A
¹⁷Are the same or different and the side chains are
The neutral amino acid residue which may be described in (113) above.
Agent,

(127) The agent according to (30) above, wherein P1 is a hydrogen atom, pGlu or Arg-Arg-Gln, and (128) P2 is an optionally substituted His or an optionally substituted Ala. The agent according to (30), the agent according to (30), wherein (129) P3 is Arg which may be substituted or Lys which may be substituted, (130) -P4-P5 is -Cha-Pro. −
Phe (Cl), -Cha-Pro-Phe, -Met
-Pro-Phe, -Met-Pro-Phe (C
l), the agent according to (30) above, which is -Cha-Phe or -Met-Phe, and the modified form of (131) apelin is Arg-Pro-Arg-Leu-Phe-Ala-Arg-Gly-Pro-Cha-. Pro-Phe
(Cl), Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Cha-Pro-Phe
(Cl), Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Cha-Pro-Ph
e, Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Met-Pro-Phe
(Cl), Arg-Pro-Arg-Leu-Phe-Ala-Arg-Gly-Pro-Met-Pro-Ph
e, Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Cha-Phe (C
l), pGlu-Arg-Pro-Arg-Leu-Phe-Arg-Arg-Gly-Pro-Met-Pro
-Phe, pGlu-Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Met-Pro
-Phe or the agent according to (30) above, which is represented by Arg-Pro-Arg-Leu-Phe-Ala-Arg-Gly-Pro-Met-Phe.

(132) The agent according to (31) above, wherein Q1 is a hydrogen atom, pGlu or Arg-Arg-Gln, and (133) Q2 is optionally substituted Arg or optionally substituted Lys. The agent according to (31) above, wherein (134) -Q3-Q4 is -OH, -Met,-.
Met-Pro-Phe, -Met-Pro-Phe
(Cl), -Cha-Pro-Phe, -Cha-Pr
o-Phe (Cl), -Ala-Pro-Phe (C
l), the agent according to (31) above, which is -Cha-Phe (Cl) or -Cha, (135) Q5 optionally substituted Pro, optionally substituted Gly or optionally substituted. The agent according to (31) above, which is Ala, and the modified form of (136) apelin is (i) Arg-Arg-Gln-Ar.
g-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Met-Pro-Phe (C
l), (ii) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Met-P
ro-Phe (Cl), (iii) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-
Pro-Cha-Phe (Cl), (iv) Arg-Pro-Arg-Leu-Ser-Ala-Arg-
Gly-Pro-Cha-Pro-Phe (Cl), (v) Arg-Arg-Gln-Arg-Pro-A
rg-Leu-Ser-Ala-Arg-Gly-Pro-Cha, (vi) Arg-Pro-Arg-L
eu-Ser-Ala-Arg-Gly-Pro-Cha-Pro-Phe, (vii) Arg-Arg-
Gln-Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Met-Pro-Ph
e, (viii) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-Ala-Arg-
Gly-Pro-Met, (ix) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-
Ala-Arg-Gly-Pro, (x) Arg-Pro-Arg-Leu-Ser-Ala-Arg-G
ly-Pro-Met-Pro-Phe, (xi) pGlu-Arg-Pro-Arg-Leu-Ser-
Ala-Arg-Gly-Pro-Met-Pro-Phe, (xii) Arg-Pro-Arg-Leu
-Ser-Ala-Arg-Gly-Pro-Met, (xiii) pGlu-Arg-Pro-Arg-
Leu-Ser-Ala-Lys-Gly-Pro-Met-Pro-Phe, (xiv) Arg-Pro
-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Ala-Pro-Phe (Cl), (xv)
Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Gly-Met-Pro-Phe (C
l), (xvi) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-NMe-Ala-
Met-Pro-Phe (Cl), (xvii) Arg-Pro-Arg-Leu-Ser-Ala-Ar
g-Gly-Pro-Cha-Pyn, (xviii) Arg-Pro-Arg-Leu-Ser-Ala
-Arg-Gly-Pro-Cha-Pro-Pyn, (xix) Arg-Arg-Gln-Arg-Pr
o-Arg-Leu-Ser-Ala-Arg-Gly-Gly-Cha, or (xx) Arg-
Pro-Arg-Leu-Ser-Ala-Lys (Me) ₂ -Gly-Pro-Met-Pro-Phe,
The agent according to (31) above, which is a peptide represented by: or an ester thereof, an amide thereof, or a salt thereof,

(137) The agent according to (32) above, wherein R1 is a hydrogen atom, pGlu or Arg-Arg-Gln, and the modified form of (138) apelin is Arg-Arg-Gln-Arg-Pro-Arg-Leu-. Ser-His-Lys-Gly-Pro-
Cha-Pro-Phe (Cl), Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Cha-Pro-Phe
(Cl), Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe
(Cl), pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Cha-Pro
-Phe, pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro
-Phe (Cl), Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-
Nle-Pro-Tyr, Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Nal
(2), pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro
-Nal (2), or Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Ch
The agent according to (32) above, which is a peptide represented by a, or its ester, its amide, or its salt, (1
39) The modified form of apelin is (i) Arg-Arg-Gln-Arg-Pro-
Arg-Leu-Ser-His-Lys-Gly-Pro-Met (O), (ii) Arg-Arg-L
ys (Arg-Arg) -Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Me
t-Pro-Phe, (iii) Arg-Arg-Arg-Arg-Pro-Arg-Leu-Ser-H
is-Lys-Gly-Pro-Met-Pro-Phe, (iv) Arg-Arg-Lys-Arg-P
ro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe, (v) Ar
g-Arg-Ala-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-
Pro-Phe, (vi) Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-
Met-Phe (Cl), (vii) pGlu-Arg-Pro-Arg-Leu-Ser-Arg-Ly
s-Gly-Pro-Met-Pro-Phe, (viii) Arg-Arg-Phe-Arg-Pro-
Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe, (ix) pGlu
-Arg-Pro-Arg-Leu-Ser-His-Arg-Gly-Pro-Met-Pro-Phe,
(x) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-P
he (Cl), (xi) Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-M
et-Cha, (xii) pGlu-Arg-Pro-Arg-Leu-Ser-Leu-Lys-Gly
-Pro-Met-Pro-Phe, (xiii) Arg-Pro-Arg-Leu-Ser-His-L
ys-Gly-Pro-Met-Nal (2), (xiv) pGlu-Arg-Pro-Arg-Leu-
Ser-Arg-Arg-Gly-Pro-Met-Pro-Phe, (xv) pGlu-Arg-Pro
-Arg-Leu-Ser-Phe-Lys-Gly-Pro-Met-Pro-Phe, (xvi) pG
lu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Cha, (x
vii) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-
Nal (2), (xviii) pGlu-Arg-Pro-Arg-Leu-Ser-His-Phe-G
ly-Pro-Met-Pro-Phe, (xix) pGlu-Arg-Pro-Arg-Leu-Ser
-His-Leu-Gly-Pro-Met-Pro-Phe, (xx) Arg-Arg-Gln-Arg
-Pro-Arg-Leu-Ser-Ala-Arg-Gly-NMe2, (xxi) Arg-Arg-G
ln-Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Mor, (xxii) Arg
-Pro-Arg-Leu-Ser-His-Ala-Gly-Pro-Cha-Pro-Phe (Cl),
(xxiii) Arg-Pro-Arg-Ala-Ser-His-Lys-Gly-Pro-Cha-Pr
o-Phe (Cl), (xxiv) Arg-Pro-Lys (Me) ₂ -Leu-Ser-Ala-Arg
-Gly-Pro-Met-Pro-Phe, (xxv) Arg-Pro-Arg-Leu-Ser-Da
p-Arg-Gly-Pro-Cha-Pro-Phe (Cl), (xxvi) Arg-Pro-Arg-
Leu-Ser-Dap (Ac) c-Arg-Gly-Pro-Cha-Pro-Phe (Cl), (xxv
ii) Arg-Pro-Arg-Leu-Ser-Dap (C6) -Arg-Gly-Pro-Cha-Pr
o-Phe (Cl), (xxviii) Arg-Pro-Arg-Leu-Ser-Dap (Adi) -A
rg-Gly-Pro-Cha-Pro-Phe (Cl), or (xxix) Arg-Pro-A
The agent according to (32) above, which is a peptide represented by la-Leu-Ser-His-Lys-Gly-Pro-Cha-Pro-Phe (Cl), or an ester thereof, an amide thereof, or a salt thereof.

In the present specification, "substantially the same" means protein activity, for example, ligand-receptor binding activity, physiological characteristics (eg, adrenocorticotropic hormone secretory regulatory action, oxytocin secretory regulatory action). And the like mean substantially the same. Amino acid substitutions, deletions, additions or insertions often do not result in a significant change in the physiological or chemical properties of the polypeptide, in which case the substituted, deleted, added or inserted polypeptide will , Would be substantially identical to those without such substitutions, deletions, additions or insertions. Substantially identical substitutions of amino acids in the amino acid sequence can be selected, for example, from other amino acids in the class to which the amino acid belongs. Examples of non-polar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine. The polar (neutral) amino acids include glycine, serine, threonine,
Examples include cysteine, tyrosine, asparagine, glutamine and the like. Examples of positively charged (basic) amino acids include arginine, lysine, and histidine.
Examples of negatively charged (acidic) amino acids include aspartic acid and glutamic acid.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION Apelin used in the present invention is
Specifically, a peptide containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 1 or a partial peptide thereof, the same as or substantially the same as the amino acid sequence represented by SEQ ID NO: 5. Partial precursor peptide containing the same amino acid sequence, Partial precursor peptide containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 7,
A precursor partial peptide containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 9, or the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 11. And a peptide containing a partial peptide of a precursor containing

The apelin of the present invention (hereinafter sometimes simply referred to as a peptide), its production method and use are described in more detail below. As apelin, any tissue of human or warm-blooded animal (eg, guinea pig, rat, mouse, pig, sheep, cow, monkey, etc.) (eg, pituitary, pancreas, brain, kidney, liver, gonad, thyroid, gallbladder, Peptide derived from bone marrow, adrenal gland, skin, muscle, lung, gastrointestinal tract, blood vessel, heart, etc. or cell and containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 Any peptide can be used, and specifically, a peptide containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 1 or a partial peptide thereof, SEQ ID NO: 5 A partial peptide of the precursor containing an amino acid sequence identical or substantially identical to the amino acid sequence represented by SEQ ID NO: 7. Precursor peptide containing the same or substantially the same amino acid sequence as the amino acid sequence, and the precursor partial peptide containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 9. , Or a partial peptide of the precursor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 11, and the like. For example, as the apelin of the present invention, in addition to the peptide containing the amino acid sequence represented by SEQ ID NO: 1, the amino acid sequence represented by SEQ ID NO: 1 is about 50 to 99.9% (preferably 70 to 99). 9.9%, more preferably 80-99.9.
%, More preferably 90 to 99.9%), and a peptide having an amino acid sequence having homology and having substantially the same activity as the peptide containing the amino acid sequence represented by SEQ ID NO: 1, SEQ ID NO: A peptide having substantially the same activity as the precursor partial peptide represented by 5
A peptide having substantially the same activity as the partial peptide of the precursor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 7, and the amino acid sequence represented by SEQ ID NO: 9 Contains a peptide having substantially the same activity as the partial peptide of the precursor containing the same or substantially the same amino acid sequence, or the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 11. And a peptide having substantially the same activity as the partial peptide of the precursor. Examples of substantially the same activity include receptor binding activity and signal transduction activity. "Substantially the same quality" means that the receptor binding activity and the like are qualitatively the same. Therefore, strength and weakness such as strength of receptor binding activity and quantitative factors such as molecular weight of peptide may be different.

Specifically as the apelin, SEQ ID NO:
The amino acid sequence represented by 1 or a partial sequence thereof,
Partial sequence of amino acid sequence represented by SEQ ID NO: 5, Partial sequence of amino acid sequence represented by SEQ ID NO: 7, Partial sequence of amino acid sequence represented by SEQ ID NO: 9, represented by SEQ ID NO: 11 Mouse brain, rat brain, which contains an amino acid sequence represented by a partial sequence of the amino acid sequence, etc.
Examples include peptides derived from pig brain, pig small intestine, bovine hypothalamus, bovine stomach, human hypothalamus or human lung. Furthermore, in the amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof, the partial sequence of the amino acid sequence represented by SEQ ID NO: 5, the partial sequence of the amino acid sequence represented by SEQ ID NO: 7, and the SEQ ID NO: 9 A partial sequence of the amino acid sequence represented, or a peptide containing the same or substantially the same amino acid sequence as the partial sequence of the amino acid sequence represented by SEQ ID NO: 11, or one or more of the partial peptides Peptides containing an amino acid sequence in which amino acids are substituted, deleted, added or inserted are included as peptides containing substantially the same amino acid sequence. For example, with the amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof, the partial sequence of the amino acid sequence represented by SEQ ID NO: 5, the partial sequence of the amino acid sequence represented by SEQ ID NO: 7, or the SEQ ID NO: 9. Partial sequence of the amino acid sequence represented, or 1 to 7 in the partial sequence of the amino acid sequence represented by SEQ ID NO: 11,
An amino acid sequence in which 1 or more and 5 or less, more preferably 1 or more and 3 or less amino acids are deleted, the amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof,
A partial sequence of the amino acid sequence represented by SEQ ID NO: 5,
1 to 20 in the partial sequence of the amino acid sequence represented by SEQ ID NO: 7, the partial sequence of the amino acid sequence represented by SEQ ID NO: 9, or the partial sequence of the amino acid sequence represented by SEQ ID NO: 11. Preferably 1 or more and 15 or less,
More preferably, an amino acid sequence in which 1 or more and 10 or less amino acids are added (or inserted), SEQ ID NO: 1
Of the amino acid sequence represented by or a partial sequence thereof, the partial sequence of the amino acid sequence represented by SEQ ID NO: 5, the partial sequence of the amino acid sequence represented by SEQ ID NO: 7, the amino acid sequence represented by SEQ ID NO: 9. Partial sequence or 1 in the partial sequence of the amino acid sequence represented by SEQ ID NO: 11
Peptides containing an amino acid sequence in which 1 or more and 7 or less, preferably 1 or more and 5 or less, more preferably 1 or more and 3 or less amino acids are substituted with other amino acids, and the like. Furthermore, the apelin or its partial peptide contains G
Also included are those in which the N-terminal side of ln is cleaved in vivo and the Gln is pyroglutamine-oxidized. The precursor of apelin may be any protein as long as it is a protein containing apelin as its partial sequence, and specifically, it contains the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11. And the like (hereinafter, the apelin and the precursor may be collectively referred to as apelin). Also,
The molecular weight of apelin is about 1000-10,000 daltons,
It is preferably about 1000 to about 5000 daltons, more preferably about 1000 to about 3000 daltons.

In the present specification, the peptides have the N-terminal (amino terminal) at the left end and the C-terminal at the right end according to the convention of peptide notation.
It is a terminal (carboxyl terminal). Amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof, partial sequence of amino acid sequence represented by SEQ ID NO: 5, partial sequence of amino acid sequence represented by SEQ ID NO: 7, represented by SEQ ID NO: 9 A peptide containing a partial sequence of the amino acid sequence represented by SEQ ID NO: 11 or the like has a C-terminal group usually having a carboxyl group (-COO
H) or carboxylate (-COO ^- is a), C-terminal may be an amide (-CONH ₂₎ or ester (-COOR). Examples of R of the ester include methyl, ethyl,
C _1-6 alkyl group such as n-propyl, isopropyl or n-butyl, C _3-8 cycloalkyl group such as cyclopentyl and cyclohexyl, C _6-12 aryl group such as phenyl and α-naphthyl, benzyl, phenethyl and benzhydryl. Such as phenyl-C _1-2 alkyl, or α-naphthyl-such as α-naphthylmethyl
In addition to C _7-14 aralkyl groups such as C _1-2 alkyl, pivaloyloxymethyl ester widely used as an oral ester can be mentioned. Amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof, partial sequence of amino acid sequence represented by SEQ ID NO: 5, partial sequence of amino acid sequence represented by SEQ ID NO: 7, represented by SEQ ID NO: 9 When a peptide containing a partial sequence of the amino acid sequence represented by SEQ ID NO: 11, or a partial sequence of the amino acid sequence represented by SEQ ID NO: 11 has a carboxyl group or a carboxylate other than at the C-terminal, those groups are amidated or Those which are esterified are also included in the peptides of the present invention. As the ester at this time, for example, the above-mentioned C-terminal ester or the like is used. As the salt of apelin, salts with physiologically acceptable bases (for example, alkali metals etc.) and acids (organic acids, inorganic acids) are used, and physiologically acceptable acid addition salts are particularly preferable. Examples of such salts include salts with inorganic acids (for example, hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid), or organic acids (for example, acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid, Salts such as tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid) are used. Apelin can be produced by a method for purifying peptides from human or warm-blooded animal tissues or cells, or can be produced according to the peptide synthesis method described below. It can also be produced by culturing a transformant containing a DNA encoding apelin described below. When producing from tissues or cells of humans or warm-blooded animals, homogenize tissues or cells of humans or warm-blooded animals, and then extract with acid, etc., and extract the extract by salting out, dialysis, gel filtration, reverse phase Purification and isolation can be achieved by combining chromatography such as chromatography, ion exchange chromatography, affinity chromatography and the like.

As described above, apelin can be produced by a peptide synthesis method known per se, or by cleaving an apelin-containing precursor with an appropriate peptidase. The peptide synthesis method may be either a solid phase synthesis method or a liquid phase synthesis method. That is, the target peptide can be produced by condensing a partial peptide or amino acid capable of forming apelin with the remaining portion and removing the protecting group when the product has a protecting group. Examples of known condensation methods and elimination of protective groups include the methods described in the following (1) to (3). M. Bodanszky and MA Ondetti, Peptide Synthesis, Interscience Publisher
s, New York (1966) Schroeder and Luebke, The Peptide,
Academic Press, New York (1965) Nobuo Izumiya et al., Fundamentals and experiments of peptide synthesis, Maruzen Co., Ltd.
(1975) Haruaki Yajima and Shunpei Sakakibara, Laboratory of Biochemistry 1, Protein Chemistry IV, 205, (1977) Supervised by Haruaki Yajima, Development of Pharmaceuticals Volume 14 Peptide Synthesis Hirokawa Shoten The peptide of the present invention can be purified and isolated by a combination of purification methods such as solvent extraction, distillation, column chromatography, liquid chromatography, recrystallization and the like. When the apelin obtained by the above method is a free form, it can be converted into a suitable salt by a known method, and conversely, when it is obtained with a salt, it can be converted into a free form by a known method. it can.

As the amide form of apelin, a commercially available resin for peptide synthesis suitable for amide formation can be used. Examples of such resin include chloromethyl resin, hydroxymethyl resin, benzhydrylamine resin, aminomethyl resin, 4-benzyloxybenzyl alcohol resin, 4-methylbenzhydrylamine resin, PAM resin,
4-hydroxymethylmethylphenylacetamidomethyl resin, polyacrylamide resin, 4- (2 ′, 4′-dimethoxyphenyl-hydroxymethyl) phenoxy resin,
4- (2 ', 4'-dimethoxyphenyl-Fmoc aminoethyl) phenoxy resin etc. can be mentioned. Using such a resin, an amino acid having an α-amino group and a side chain functional group appropriately protected, according to the sequence of the target peptide,
The condensation is performed on the resin according to various condensation methods known per se.
At the end of the reaction, the peptide is cleaved from the resin and at the same time various protective groups are removed, and if necessary, an intramolecular disulfide bond forming reaction is carried out in a highly diluted solution to obtain the target peptide. Regarding the above-mentioned condensation of protected amino acids, various activating reagents that can be used for peptide synthesis can be used, but carbodiimides are particularly preferable. Carbodiimides include DCC, N, N'-diisopropylcarbodiimide, N-ethyl-N '-(3-dimethylaminopropyl)
Carbodiimide etc. are mentioned. For activation by these, the protected amino acid is added directly to the resin with a racemization inhibitor additive (e.g., HOBt, HOOBt, etc.) or the amino acid previously protected as a symmetrical acid anhydride or HOBt ester or HOOBt ester is It can be added to the resin after activation. The solvent used for activation of the protected amino acid or condensation with the resin can be appropriately selected from solvents known to be usable for peptide condensation reaction. For example, N, N-dimethylformamide, N, N-dimethylacetamide, N-
Acid amides such as methylpyrrolidone, halogenated hydrocarbons such as methylene chloride and chloroform, alcohols such as trifluoroethanol, sulfoxides such as dimethyl sulfoxide, tertiary amines such as pyridine,
Ethers such as dioxane and tetrahydrofuran, nitriles such as acetonitrile and propionitrile, esters such as methyl acetate and ethyl acetate, and appropriate mixtures thereof are used. The reaction temperature is appropriately selected from the range known to be applicable to peptide bond-forming reactions, and is usually selected in the range of approximately -20 ° C to 50 ° C. The activated amino acid derivative is usually 1.
Used in 5- to 4-fold excess. As a result of the test using the ninhydrin reaction, when the condensation is insufficient, sufficient condensation can be performed by repeating the condensation reaction without removing the protecting group. When sufficient condensation cannot be obtained even after repeating the reaction, the unreacted amino acid can be acetylated with acetic anhydride or acetylimidazole so as not to affect the subsequent reaction. Examples of the protecting group for the amino group of the raw material amino acid include, for example,
Z, Boc, tertiary pentyloxycarbonyl, isobornyloxycarbonyl, 4-methoxybenzyloxycarbonyl, Cl-Z, Br-Z, adamantyloxycarbonyl, trifluoroacetyl, phthaloyl, formyl, 2-nitrophenylsulfenyl, Examples include diphenylphosphinothioyl oil and Fmoc. Examples of the protecting group for the carboxyl group include C described above as R.
_1-6 alkyl group, C _3-8 cycloalkyl group, C
_{In addition to the 7-14} aralkyl group, 2-adamantyl, 4-nitrobenzyl, 4-methoxybenzyl, 4-chlorobenzyl, phenacyl group and benzyloxycarbonyl hydrazide, tertiary butoxycarbonyl hydrazide, trityl hydrazide and the like can be mentioned. The hydroxyl groups of serine and threonine can be protected by, for example, esterification or etherification. Examples of groups suitable for this esterification include lower alkanoyl groups such as acetyl group, aroyl groups such as benzoyl group, groups derived from carbon such as benzyloxycarbonyl group and ethoxycarbonyl group. Examples of the group suitable for etherification include a benzyl group, a tetrahydropyranyl group and a tert-butyl group. Examples of the protective group for the phenolic hydroxyl group of tyrosine include, for example,
Bzl, Cl ₂ -Bzl, 2-nitrobenzyl, Br-Z, tertiary butyl and the like can be mentioned. Examples of the protecting group for imidazole of histidine include Tos, 4-methoxy-2,3,6-trimethylbenzenesulfonyl, DNP, benzyloxymethyl, Bum, Boc, Trt, Fmoc and the like. Examples of the activated carboxyl group of the raw material include corresponding acid anhydrides, azides, active esters [alcohols (for example, pentachlorophenol, 2,4,5-trichlorophenol, 2,4-dinitrophenol, cyano Methyl alcohol, para-nitrophenol, HONB, N-hydroxysuccinimide, N-hydroxyphthalimide, ester with HOBt)] and the like. The activated amino group of the raw material includes, for example, the corresponding phosphoric acid amide.

As a method of removing (eliminating) the protecting group, for example, catalytic reduction in a hydrogen stream in the presence of a catalyst such as Pd black or Pd carbon, or anhydrous hydrogen fluoride, methanesulfonic acid or trifluoride is used. Acid treatment with methanesulfonic acid, trifluoroacetic acid, or a mixture thereof, base treatment with diisopropylethylamine, triethylamine, piperidine, piperazine, etc., reduction with sodium in liquid ammonia, and the like are also included. The elimination reaction by the acid treatment is generally carried out at a temperature of -20 ° C to 40 ° C, but in the acid treatment, anisole, phenol, thioanisole, metacresol, paracresol, dimethyl sulfide, 1,4-butanedithiol, 1 , 2-
It is effective to add a cation trapping agent such as ethanedithiol. Also, the 2,4-dinitrophenyl group used as the imidazole protecting group of histidine is removed by thiophenol treatment, and the formyl group used as the indole protecting group of tryptophan is the above 1,2-ethanedithiol, 1,4-butane group. In addition to deprotection by acid treatment in the presence of dithiol or the like, it is also removed by alkali treatment with dilute sodium hydroxide or dilute ammonia. Protection of a functional group which should not be involved in the reaction of the raw material, protection of the protective group, elimination of the protective group, activation of the functional group involved in the reaction and the like can be appropriately selected from known groups or known means. As another method for obtaining the amide form of apelin, first, after amidating the α-carboxyl group of the carboxyl terminal amino acid, the peptide chain is extended to the amino group side to the desired chain length, and then the N-terminal of the peptide chain And a peptide (or amino acid) from which only the protecting group for the C-terminal carboxyl group has been removed, and both peptides are condensed in the mixed solvent as described above. . The details of the condensation reaction are the same as above. After purifying the protected peptide obtained by condensation,
All the protecting groups can be removed by the above method to obtain the desired crude peptide. This crude peptide can be purified by making full use of various known purification means, and the main fraction can be lyophilized to obtain the amide form of the desired peptide. In order to obtain an ester form of apelin, the α-carboxyl group of a carboxy terminal amino acid is condensed with a desired alcohol to form an amino acid ester, and then the desired ester form of apelin can be obtained in the same manner as the amide form of the peptide.

As apelin, the above-mentioned SEQ ID NO:
Amino acid sequence represented by 1 or a partial sequence thereof, partial sequence of amino acid sequence represented by SEQ ID NO: 5, partial sequence of amino acid sequence represented by SEQ ID NO: 7, amino acid sequence represented by SEQ ID NO: 9 Or a partial sequence of the amino acid sequence represented by SEQ ID NO: 11, or the like, which has the same or substantially the same amino acid sequence as the peptide (eg, adrenocorticotropic hormone secretory regulatory action, oxytocin secretion) Any peptide may be used as long as it has a regulatory action). Examples of such a peptide include, for example, the amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof,
A partial sequence of the amino acid sequence represented by SEQ ID NO: 5,
One or more from a peptide having a partial sequence of the amino acid sequence represented by SEQ ID NO: 7, a partial sequence of the amino acid sequence represented by SEQ ID NO: 9, or a partial sequence of the amino acid sequence represented by SEQ ID NO: 11 A peptide having an amino acid sequence in which the amino acid of is deleted.
Specifically, (1) a peptide having the first to twelfth amino acid sequences of the amino acid sequence represented by SEQ ID NO: 1, and (2) the first amino acid sequence represented by SEQ ID NO: 1.
A peptide having the thirteenth to thirteenth amino acid sequence,
(3) A peptide having the 1st to 14th amino acid sequences of the amino acid sequence represented by SEQ ID NO: 1, (4) 1st to 1st of the amino acid sequence represented by SEQ ID NO: 1
A peptide having the 5th amino acid sequence, (5) a peptide having the 1st to 16th amino acid sequences of the amino acid sequence represented by SEQ ID NO: 1, (6) SEQ ID NO:
Peptides having a partial sequence of the amino acid sequence represented by 5, 7, 9 or 11 are preferable. Of these, peptides having a partial sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 are preferable.

The peptide having the partial sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 is specifically (a) SEQ ID NO: 5, 7, 9 or 11
A peptide having the 6th to 77th amino acid sequences of the amino acid sequence represented by: (b) SEQ ID NO: 5,
A peptide having the 40th to 77th amino acid sequences of the amino acid sequence represented by 7, 9 or 11;
(C) a peptide having an amino acid sequence from the 42nd position to the 77th position in the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (d) SEQ ID NO: 5, 7, 9 or 1
A peptide having the 47th to 77th amino acid sequence of the amino acid sequence represented by 1, (e) SEQ ID NO:
The 61st of the amino acid sequence represented by 5, 7, 9 or 11
A peptide having the amino acid sequence from the 77th to the 77th,
(F) A peptide having the 65th to 77th amino acid sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 or its N-terminal amino acid (Gln)
Which was oxidized with pyroglutamine, (g) SEQ ID NO: 5,
A peptide having the 1st to 25th amino acid sequences of the amino acid sequence represented by 7, 9 or 11 (h)
A peptide having the 6th to 25th amino acid sequences of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (i) the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 A peptide having the 42nd to 64th amino acid sequence of (b), (j) SEQ ID NO: 5, 7, 9
Or a peptide having the 61st to 64th amino acid sequence of the amino acid sequence represented by 11, and (k) the 43rd to 77th amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11. A peptide having the amino acid sequence of (1), a peptide having the 41st to 77th amino acid sequence of the amino acid sequence represented by SEQ ID NOs: 5, 7, 9 or 11, (m) the SEQ ID NOs: 5, 7 66th to 7th of the amino acid sequence represented by 9 or 11
Peptide having the 7th amino acid sequence, (n) Peptide having the 67th to 77th amino acid sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (o) SEQ ID NO: 5 , A peptide having the 64th to 77th amino acid sequence of the amino acid sequence represented by 7, 9 or 11, (p) the 63rd amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 From 77th
A peptide having the amino acid sequence of the third position, (q) a peptide having the amino acid sequence of the 65th to the 76th position in the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (r) the SEQ ID NO: 5, Peptides having the 65th to 75th amino acid sequences of the amino acid sequence represented by 7, 9 or 11 are mentioned, among which the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 A peptide having the 65th to 77th amino acid sequence, or an amino acid (Gln) at the N-terminal thereof that has been pyroglutamine-oxidized or SEQ ID NO: 5, 7, 9 or 1
A peptide having the 42nd to 77th amino acid sequences of the amino acid sequence represented by 1 is preferably used. In particular, the peptide represented by the 65th to 77th amino acid sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 or the N-terminal amino acid (Gln) thereof is pyroglutamine-oxidized (pGlu Arg P
ro Arg Leu Ser His Lys Gly Pro Met Pro Phe) is preferred. In addition pGlu Arg Pro Arg Leu Ser His Lys Gly P
A peptide having a partial amino acid sequence of the amino acid sequence represented by ro Met Pro Phe is also preferably used as apelin.

The partial peptide of apelin can be further used as an antigen for preparing an anti-apelin antibody.
In addition to the above-mentioned apelin, partial peptides such as N-terminal peptide, C-terminal peptide and central peptide of the above-mentioned apelin are used as the peptide as such an antigen. As the partial peptide, a peptide containing individual domains may be used, but a partial peptide containing a plurality of domains at the same time may be used. The partial peptide in the present specification also has an amide (-CONH ₂ ) or ester (-CO at the C-terminal).
OR). Here, examples of the ester group are the same as those in the above-mentioned peptide. When the partial peptide has a carboxyl group or a carboxylate in addition to the C-terminal, those having an amidated or esterified group are also included in the partial peptide of the present invention.
As the ester at this time, for example, the above-mentioned C-terminal ester is used. Apelin or its partial peptide may be a fusion protein with a protein whose function or property is well known. As the salt of the partial peptide of apelin, the same salts as the above-mentioned salts of apelin are used. The partial peptide of apelin or its amide, ester or salt thereof can be produced according to the same synthetic method as in the case of the above-mentioned apelin or by cleaving apelin with an appropriate peptidase.

As the DNA encoding apelin, specifically, the above-mentioned amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof, partial sequence of the amino acid sequence represented by SEQ ID NO: 5, SEQ ID NO: The same as or substantially the same as the partial sequence of the amino acid sequence represented by 7, the partial sequence of the amino acid sequence represented by SEQ ID NO: 9, or the partial sequence of the amino acid sequence represented by SEQ ID NO: 11. Any DNA can be used as long as it contains a DNA encoding an apelin containing the same amino acid sequence. Also, genomic DNA, genome D
NA library, tissue- or cell-derived cDNA described above
A, the above-mentioned tissue / cell-derived cDNA library,
Any of synthetic DNA may be used. The vector used for the library may be any of bacteriophage, plasmid, cosmid, phagemid and the like. In addition, reverse Transcriptase Polymerase Chain React was directly performed using the RNA fraction prepared from the tissues and cells described above.
It can also be amplified by ion (hereinafter, abbreviated as RT-PCR method). More specifically, the amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof, the partial sequence of the amino acid sequence represented by SEQ ID NO: 5, the partial sequence of the amino acid sequence represented by SEQ ID NO: 7, a sequence Mouse whole brain, rat whole brain, bovine hypothalamus, bovine stomach derived, human hypothalamus containing the partial sequence of the amino acid sequence represented by NO: 9 or the partial sequence of SEQ ID NO: 11 or The DNA encoding a human lung-derived polypeptide includes (1) a base sequence represented by SEQ ID NO: 2 or a partial sequence thereof, a partial sequence of the base sequence represented by SEQ ID NO: 6, and a sequence represented by SEQ ID NO: 8. Containing a DNA having a partial sequence of the base sequence represented by SEQ ID NO: 10, a partial sequence of the base sequence represented by SEQ ID NO: 10, or a partial sequence of the base sequence represented by SEQ ID NO: 12.
(2) a mammalian-derived DNA that hybridizes with the sequence defined in (1) under stringent conditions, (3) the sequence defined in (1) and (2) due to the degeneracy of the genetic code A DNA that does not hybridize with, but encodes a polypeptide having the same amino acid sequence is used. Hybridization can be performed according to a method known per se or a method analogous thereto. The stringent conditions are, for example, 42 ° C., 50% formamide, 4 × SSPE (1 × SSPE = 150 mM NaCl, 10 mM
_{NaH 2 PO 4 · H 2 O} , 1mM EDTA pH7.4), 5 × Denhardt's solution and 0.1% SDS. In the base sequence represented by SEQ ID NO: 2, Y is T or C, N is T, C,
A or G, R is A or G, M is C or A,
W represents T or A, and S represents C or G. In addition, the amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof, the partial sequence of the amino acid sequence represented by SEQ ID NO: 5, the partial sequence of the amino acid sequence represented by SEQ ID NO: 7, and the SEQ ID NO: 9 For example, 6 or more and 51 or less (preferably 9 or more and 3 or less) in the DNA encoding the polypeptide containing the partial sequence of the represented amino acid sequence or the partial sequence of the amino acid sequence represented by SEQ ID NO: 11.
A DNA fragment containing a partial base sequence of 0 or less, more preferably 12 or more and 30 or less) is also preferably used as a DNA detection probe.

The DNA encoding apelin can also be produced by the following genetic engineering method. As a means for cloning a DNA that completely encodes apelin, the above DNA can be prepared by a PCR method known per se using a synthetic DNA primer having a partial base sequence of apelin.
The target DNA is amplified from a library or the like, or the DNA inserted into an appropriate vector is labeled with, for example, a DNA fragment having a partial or entire region of the polypeptide of the present invention or a synthetic DNA. It can be selected by hybridization. Hybridization methods include, for example, Molecular Cloni
ng (2nd ed .; J. Sambrook et al., Cold Spring Harbo
r Lab. Press, 1989). When a commercially available library is used, the method described in the attached instruction manual is used. Depending on the purpose, the cloned DNA encoding apelin is
Alternatively, it can be used after digestion with a restriction enzyme or addition of a linker, if desired. The DNA has ATG as a translation initiation codon at the 5'terminal side, and TAA and TG as translation termination codons at the 3'terminal side.
It may have A or TAG. These translation initiation codon and translation termination codon can be added using an appropriate synthetic DNA adaptor. An apelin expression vector can be produced, for example, by (a) cutting out a target DNA fragment from apelin-encoding DNA, and (b) ligating the DNA fragment downstream of a promoter in a suitable expression vector. it can. As the vector, a plasmid derived from E. coli (eg, pBR322, pB
R325, pUC12, pUC13), a Bacillus subtilis-derived plasmid (eg, pUB110, pTP5, pC19)
4), yeast-derived plasmid (eg, pSH19, pSH1
5), bacteriophages such as λ phage, animal viruses such as retrovirus, vaccinia virus, baculovirus, etc. are used. The promoter used in the present invention may be any promoter as long as it is suitable for the host used for gene expression.

When the host used for transformation is an animal cell, SV40-derived promoter, retrovirus promoter, metallothionein promoter, heat shock promoter, cytomegalovirus promoter, SRα promoter and the like can be used. When the host is Escherichia, the trp promoter,
T7 promoter, lac promoter, recA promoter, .lambda.P _L promoter, lpp promoter, etc. When the host is Bacillus, SPO1 promoter, SPO2 promoter, etc. When the host is yeast penP promoter, PHO5 promoter, PGK Promoter, GAP promoter, ADH
1 promoter, GAL promoter and the like are preferable.
When the host is an insect cell, polyhedrin promoter, P10 promoter and the like are preferable. In addition to the above, the expression vector may optionally include an enhancer, a splicing signal, a poly A addition signal, a selection marker,
Those containing an SV40 origin of replication (hereinafter sometimes abbreviated as SV40ori) and the like can be used. Examples of the selection marker include dihydrofolate reductase (hereinafter sometimes abbreviated as dhfr) gene [methotrexate (MTX) resistance], ampicillin resistance gene (hereinafter sometimes referred to as Amp ^r ),
Neomycin resistance gene (hereinafter sometimes referred to as Neo, G418 resistance) and the like can be mentioned. Especially CHO
When the DHFR gene is used as a selection marker using (dhfr ⁻ ) cells, selection can also be performed using a thymidine-free medium. If necessary, a signal sequence suitable for the host is added to the N-terminal side of the polypeptide or its partial peptide. When the host is Escherichia, the phoA signal sequence, OmpA signal sequence and the like, when the host is Bacillus α-amylase signal sequence, subtilisin signal sequence, etc., the host is yeast In some cases, the mating factor α (MFα) signal sequence, invertase signal sequence, etc. When the host is an animal cell, for example, insulin signal sequence, α-interferon signal sequence, antibody molecule, signal sequence, etc. Can be used respectively. A transformant can be produced using the vector containing the DNA encoding the polypeptide thus constructed.

As the host, for example, Escherichia, Bacillus, yeast, insects or insect cells, animal cells, etc. are used. Examples of Escherichia bacteria include Escherichia coli K12 and DH1.
[Proc. Natl. Acad. Sci. USA], Volume 60, 160
(1968)], JM103 [Nukuirek Acids.
Research (Nucleic Acids Research), Volume 9, 309
(1981)], JA221 [Journal of Molecular Biolog (Journal of Molecular Biolog
y)], Volume 120, 517 (1978)], HB101
[Journal of Molecular Biology, 4
1 volume, 459 (1969)], C600 [Genetics, 39 volume, 440 (1954)] and the like. Examples of Bacillus include Bacillus
Bacillus subtilis MI114 [Gene,
24, 255 (1983)], 207-21 [Journal of Biochemis
try), Vol. 95, 87 (1984)] and the like are used.

Examples of yeast include Saccaromyces cerevisiae AH22,
^{AH22R -, NA87-11A, DKD-5D} , 20
B-12 or the like is used. Examples of insects include silkworm larvae [Maeda et al., Nature (Natu
re), 315, 592 (1985)]. Examples of insect cells include Spodoptera frugiperda cell (Sf) when the virus is AcNPV.
Cells), MG1 cells derived from the midgut of Trichoplusia ni, Tri
High Five ^TM cells derived from choplusia ni eggs, Mamestrab
Cells derived from rassicae or cells derived from Estigmena acrea are used. When the virus is BmNPV, silkworm-derived cell lines (Bombyx mori N; BmN cells) and the like are used. Examples of the Sf cells include Sf9 cells (ATCC CRL1711), Sf21 cells [above, Vaughn, JL
Et al., In Vitro, Volume 13, 213-2
17 (1977)] and the like are used. Examples of animal cells include monkey COS-7 cells, Vero cells, Chinese hamster cells CHO, DHFR gene-deficient Chinese hamster cells CHO (dhfr ^- CHO).
Cells), mouse L cells, mouse 3T3 cells, mouse myeloma cells, human HEK293 cells, human FL cells, 2
93 cells, C127 cells, BALB3T3 cells, Sp-
2 / O cells and the like are used. To transform Escherichia, for example, Procedures of the
National Academy of Sciences of the USA (Proc. Natl. Acad. Sci. US
A), Volume 69, 2110 (1972) and Gene,
Vol. 17, 107 (1982) and the like. To transform Bacillus, for example, Molecular & General Genetics, 168, 11
1 (1979) and the like. To transform yeast, for example, Procedures of the National Academy of Sciences of the USA (Proc. Natl. Acad. Sci. U.
SA), Vol. 75, 1929 (1978).

To transform insect cells or insects,
For example, Bio / Technology, 6
Vol., Pp. 47-55 (1988) and the like. For transforming animal cells, for example, Virology, 52, 456 (197).
It is performed according to the method described in 3). Examples of the method for introducing an expression vector into cells include, for example, the lipofection method [Felgner, PL et al.
The National Academy of Sciences
Proceedings of the Natinal
Academy of Sciences of the United States of Americ
a), 84, 7413 (1987)], calcium phosphate method [Graham, FL and van der Eb, AJ Virology, 52, 456-467 (1).
973)], electroporation [Nuemann, E. et al. EMBO J., Vol. 1, pp. 841-845 (1982)] and the like. Thus, a transformant transformed with the expression vector containing the DNA encoding the polypeptide of the present invention can be obtained. As a method of stably expressing the polypeptide of the present invention using animal cells, there is a method of clonally selecting cells in which the expression vector introduced into the above-mentioned animal cells is integrated into the chromosome. Specifically, transformants are selected using the above selection marker as an index. Furthermore, by repeatedly performing clonal selection on the animal cells thus obtained using the selection marker, a stable animal cell line having a high expression ability of the polypeptide of the present invention or the like can be obtained. When the dhfr gene is used as a selection marker, the MTX concentration is gradually increased and the cells are cultured to select a resistant strain, so that the DNA encoding the polypeptide of the present invention or a partial peptide thereof can be introduced into cells together with the dhfr gene. It is also possible to obtain a highly expressed animal cell line by amplifying in-house. To produce the polypeptide or the like of the present invention by culturing the above transformant under conditions capable of expressing the DNA encoding the polypeptide or the like of the present invention, and producing and accumulating the polypeptide or the like of the present invention You can Host is Escherichia,
When culturing a transformant that is a bacterium of the genus Bacillus, a liquid medium is suitable as a medium used for the culture, and among them, a carbon source, a nitrogen source, an inorganic substance and the like necessary for the growth of the transformant are contained. It is included. Examples of carbon sources include glucose, dextrin, soluble starch, sucrose, etc.
As the nitrogen source, for example, ammonium salts, nitrates, corn steep liquor, peptone, casein, meat extract, soybean meal, potato extract and other inorganic or organic substances, as inorganic substances such as calcium chloride, sodium dihydrogen phosphate, Magnesium chloride etc. are mentioned. In addition, yeast, vitamins, growth promoting factors and the like may be added. The pH of the medium is preferably about 5-8.

As a medium for culturing Escherichia, for example, M9 medium containing glucose and casamino acid [Miller, Journal of Experiments in Molecular Genetics (Journa
l of Experiments in Molecular Genetics), 431-
433, Cold Spring Harbor Laboratory, New York 1
972] is preferable. If necessary, a drug such as 3β-indolylacrylic acid can be added to the promoter in order to work it efficiently. When the host is a bacterium belonging to the genus Escherichia, the culture is usually carried out at about 15 to 43 ° C. for about 3 to 24 hours, and if necessary, aeration and stirring can be added. When the host is a bacterium of the genus Bacillus, the culture is usually carried out at about 30 to 40 ° C. for about 6 to 24 hours, and aeration and stirring can be added if necessary. When culturing a transformant whose host is yeast, examples of the medium include Burkholder's minimum medium [Bostian, KL et al.
Procedures of the National Academy of Sciences of the USA (Pr
oc. Natl. Acad. Sci. USA), 77, 4505 (1)
980)] or an SD medium containing 0.5% casamino acid [Bi
tter, GA et al., The Procedures of the National Academy of Sciences of the
USA (Proc. Natl. Acad. Sci. USA), 8
1, 5330 (1984)]. P of medium
H is preferably adjusted to about 5-8. Culture is usually about 2
It is carried out at 0 ° C to 35 ° C for about 24 to 72 hours, and aeration and stirring are added if necessary.

When culturing a transformant whose host is an insect cell, Grace's Insect Medium (Grace,
TCC, Nature, 195, 788 (1962)) to which an additive such as immobilized 10% bovine serum is appropriately added may be used. The pH of the medium is preferably adjusted to about 6.2 to 6.4. Culturing is usually performed at about 27 ° C. for about 3 to 5 days, and aeration and agitation are added if necessary. When culturing a transformant whose host is an animal cell, the medium is, for example, MEM medium containing about 5 to 20% fetal bovine serum [Seience, Volume 122, 501 (1952)], D.
MEM medium [Virology, Volume 8, 396
(1959)], RPMI 1640 medium [Journal
The Jounal of the American Medical Association
199, 519 (1967)], 199 medium [Proceeding of the Society fo (Proceeding of the Society fo)
r the Biological Medicine), 73, 1 (195
0)] is used. The pH is preferably about 6-8. Culturing is usually carried out at about 30 ° C to 40 ° C for about 15 to 60 hours, and aeration and agitation are added if necessary. Especially CHO
When (dhfr ⁻ ) cells and the dhfr gene are used as selection markers, it is preferable to use DMEM medium containing dialyzed fetal bovine serum containing almost no thymidine.

The polypeptide of the present invention can be separated and purified from the above-mentioned culture by the following method, for example. When the polypeptide of the present invention is extracted from cultured bacterial cells or cells, after the culture, the bacterial cells or cells are collected by a known method, suspended in an appropriate buffer, and then subjected to ultrasonic waves, lysozyme and / or freeze-thawing. After destroying the bacterial cells or cells by, for example, a method of obtaining a crude polypeptide extract by centrifugation or filtration can be appropriately used. A protein denaturing agent such as urea or guanidine hydrochloride or Triton X-100 (registered trademark; hereinafter TM
May be omitted. ) Or the like may be included. When the polypeptide is secreted into the culture medium, after the culture is completed, the bacterial cells or cells are separated from the supernatant by a method known per se, and the supernatant is collected. Purification of the polypeptide of the present invention contained in the thus obtained culture supernatant or the extract can be carried out by appropriately combining separation / purification methods known per se. As the known separation and purification methods thereof, a method utilizing solubility such as salting out or solvent precipitation method, a dialysis method, an ultrafiltration method, a gel filtration method, and an SDS-polyacrylamide gel electrophoresis method are mainly used. The difference in charge, the difference in charge such as ion exchange chromatography, the method using specific affinity such as affinity chromatography, and the difference in hydrophobicity such as reversed-phase high performance liquid chromatography Methods such as isoelectric focusing, chromatofocusing, and other methods that utilize differences in isoelectric points are used. When the polypeptide of the present invention thus obtained is obtained in a free form, it can be converted into a salt by a method known per se or a method analogous thereto, and conversely, when it is obtained as a salt, a method known per se. Alternatively, it can be converted into a free form or another salt by a method similar thereto. In addition, the polypeptide of the present invention produced by the recombinant can be optionally modified or partially removed by acting an appropriate protein-modifying enzyme before or after purification. Examples of the protein-modifying enzyme include trypsin, chymotrypsin, arginyl endopeptidase, protein kinase, glycosidase and the like. The presence of the thus-generated polypeptide of the present invention can be measured by an enzyme immunoassay using a specific antibody.

Next, a modified form of apelin will be described.
It In the present specification, an amino acid residue means an amino acid
It loses water molecules and forms peptide bonds, which allow proteins and peptides to
Other than N-terminal or C-terminal when incorporated into peptide
The structure of a part. For example, α-amino acid residue is α-
Amino acid (H_TwoNC (R⁰) (R¹) COOH: R⁰And R¹Are each
The same or different, representing any substituent)
Lost to form peptide bonds in proteins and peptides
-N at the part other than N-terminal or C-terminal when incorporated into
HC (R⁰) (R¹) Refers to the structure of CO-. On the other hand, the N-terminal amino acid
Residue is H_TwoNC (R⁰) (R¹) CO-, C-terminal amino acid residue is -NHC
(R⁰) (R ¹) COOH. In addition, β-amino acid (H_TwoNC
(R⁰) (R¹) C (R²) (R³) COOH: R⁰, R¹, R²And R³Haso
Each is the same or different and represents an arbitrary substituent) is water
Losing molecules to form peptide bonds,
Portion other than N-terminal or C-terminal when incorporated into
-NHC (R⁰) (R¹) C (R²) (R³) Refers to the structure of CO-. On the other hand, N end
The amino acid residue at the end is H_TwoNC (R⁰) (R¹) C (R²) (R³) CO-, C end
The terminal amino acid residue is -NHC (R⁰) (R¹) C (R²) (R³) Indicated by COOH
Be done. In addition, γ-amino acid (H_TwoNC (R⁰) (R¹) C (R²) (R³) C
(R^Four) (R^Five) COOH: R⁰, R¹, R², R³, R^FourAnd R⁵Haso
Each is the same or different and represents an arbitrary substituent) is water
Losing molecules to form peptide bonds,
Portion other than N-terminal or C-terminal when incorporated into
-NHC (R⁰) (R¹) C (R²) (R³) C (R^Four) (R^Five) Refers to the structure of CO-.
On the other hand, the N-terminal amino acid residue is H _TwoNC (R⁰) (R¹) C (R²) (R³)
C (R^Four) (R^Five) CO-, C-terminal amino acid residue is -NHC (R⁰) (R¹) C
(R ²) (R³) C (R^Four) (R^Five) COOH. Furthermore, ε-ami
No acid (H_TwoNC (R⁰) (R¹) C (R²) (R³) C (R^Four) (R^Five) C (R⁶) (R⁷) C
(R⁸) (R⁹) COOH: R⁰, R¹, R², R³, R^Four, R^Five, R⁶, R⁷, R
⁸And R⁹Are the same or different and are any substitutions
Group (which represents a group) loses a water molecule to form a peptide bond.
N-terminal when incorporated into protein or peptide or
-NHC (R⁰) (R¹) C (R²) (R³) C (R^Four) (R^Five) C (R
⁶) (R⁷) C (R⁸) (R⁹) Refers to the structure of CO-. On the other hand, the N-terminal amino acid
No acid residue is H_TwoNC (R⁰) (R¹) C (R²) (R³) C (R^Four) (R^Five) C (R⁶)
(R⁷) C (R⁸) (R⁹) CO-, C-terminal amino acid residue is -NHC (R⁰) (R
¹) C (R²) (R ³) C (R^Four) (R^Five) C (R⁶) (R⁷) C (R⁸) (R⁹) Indicated by COOH
Be done.

In the present specification, an amino acid is a natural or unnatural amino acid, which is D-form or L-form.
The body may be any one of α-, β-, γ-, and ε-types. α-amino acid, β-amino acid, γ-
Examples of the group forming the side chain of amino acid and ε-amino acid include:
For example, (1) C _1-6 alkyl group (eg, methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl etc., preferably C _1-3 alkyl etc.), (2) cyano group, (3) halogen (eg fluorine, Chlorine, bromine, iodine, etc.), (4) hydroxy-C _1-6 alkyl group (eg, hydroxymethyl, hydroxyethyl, etc.), (5) C _1-6
Alkoxy group (eg, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy and the like, preferably C _1-3 alkoxy and the like), (6) C
_1-6 alkoxy-carbonyl group (eg, methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, tert-butoxycarbonyl, etc., preferably C
_1-3 alkoxy-carbonyl, etc.), (7) C _1-4 acyl group (eg, formyl such as formyl, acetyl, propionyl, butyryl and C _2-4 alkanoyl),
(8) Hydroxy group, (9) Formula: —S (O) a—R ²¹ (wherein
a is an integer of 0 to 2 and R ²¹ is a group (eg, methylthio, methanesulfinyl, methanesulfonyl, ethylthio) represented by C _1-6 alkyl (specific examples thereof are the same as those mentioned above). , Ethanesulfinyl, ethanesulfonyl, etc.), (10) benzyloxycarbonyl,
(11) tosyl group, (12) carbamoyl group, (13) mercapto group, (14) amino group, (15) sulfo group, (16) phosphono group, (1
7) phospho group, (18) carboxyl group, (19) tetrazolyl group, (20) amino-C _1-6 alkyl group (eg, aminomethyl, aminoethyl, etc.), (21) aminoallyl group, (22)
Thiazolyl group, (23) thienyl group, (24) oxazolyl group,
(25) Furyl group, (26) pyranyl group, (27) pyridyl group, (28)
Pyrazyl group, (29) pyrazinyl group, (30) pyrimidinyl group,
(31) pyridazinyl group, (32) indolyl group, (33) indodinyl group, (34) isoindolyl group, (35) pyrrolyl group, (3
6) imidazolyl group, (37) isothiazolyl group, (38) pyrazolyl group, (39) chromenyl group, (40) purinyl group, (41) quinolidinyl group, (42) quinolyl group, (43) isoquinolyl group,
(44) quinazolinyl group, (45) quinoxalinyl group, (46) cinnolinyl group, (47) morpholinyl group, (48) benzothienyl group, (49) benzofuranyl group, (50) benzimidazolyl, (51) benzimidazolyl group Group, (52) C _3-8 cycloalkyl group, (53) C substituted with the substituent described in the above (2), (3), (5) to (17), (20) to (52) _1-4 alkyl group, (54) above (2), (3), (5) to (17), (20) to (52)
Formyl substituted with the substituent described in 1., a C _1-4 acyl group such as C _2-4 alkanoyl, (55) above (1) to (52)
C _6-10 such as phenyl substituted with the substituents described in
Aryl group (mesityl, tolyl, xylyl, styrenyl, etc.), (56) C _7-15 aralkyl group such as benzyl substituted with the substituent described in the above (1) to (52) (methylbenzyl, methoxybenzyl, etc.) , (57) C _7-15 aralkyl group (benzyl, phenethyl, benzhydryl, naphthylmethyl, _etc.), (58) _{C 6- 10} aryl group (phenyl,
Naphthyl, indenyl, etc.), and (59) hydrogen atom.

A side chain forming an amino acid residue may be bonded to a nitrogen atom to form a ring (eg, proline, etc.), and two side chains may be bonded to form a ring (eg, proline). Three
-Aminonorbornanecarboxylic acid, etc.). Examples of α-amino acids include glycine, alanine, valine,
Leucine, isoleucine, serine, threonine, cysteine, methionine, aspartic acid, glutamic acid, lysine, arginine, phenylalanine, tyrosine, histidine, tryptophan, asparagine, glutamine,
Proline, pipecolic acid, norleucine, γ-methylleucine, tert-leucine, norvaline, homoarginine, homoserine, α-aminoisobutyric acid, α-aminobutyric acid, ornithine, α-aminoadipic acid, phenylglycine, thienylglycine, cyclohexylglycine, Cyclohexylalanine, thienylalanine, naphthylalanine, biphenylalanine, p-phosphonomethylphenylalanine, octahydroindole-2-carboxylic acid, O-phosphotyrosine, adamantylalanine, benzothienylalanine, pyridylalanine, piperidylalanine, pyrazylalanine, quino Lylalanine, thiazolylalanine, homocysteine, homophenylalanine, citrulline, homocitrulline, oxyproline (hydroxyproline), α, β Diaminopropionic acid,
α, γ-diaminobutyric acid, aminomalonic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, penicillamine,
Examples thereof include cycloleucine and 2-amino-4-pentenoic acid. Examples of β-amino acids include, for example, β-alanine, β-aminobutyric acid, isoasparagine, 3-aminoadipic acid, 3-aminophenylpropionic acid, 3-amino-2-
Examples thereof include hydroxy-4-phenylbutyric acid, 3-aminonorbornanecarboxylic acid, and 3-aminobicycloheptanecarboxylic acid. Examples of γ-amino acids include, for example, γ-
Aminobutyric acid, isoglutamine, statin, 4-amino-3-
Hydroxy-5-cyclohexylpentanoic acid, 4-amino-3-
Examples thereof include hydroxy-5-phenylpentanoic acid, 6-aminopenicillanic acid and 3-aminoadamantane-1-carboxylic acid. Examples of ε-amino acids include ε-aminocaproic acid, 4-aminomethyl-cyclohexanecarboxylic acid and the like.

Examples of the natural amino acids include glycine and a
Lanine, valine, leucine, isoleucine, serine, su
Leonine, cysteine, cystine, methionine, aspa
Laginic acid, Glutamic acid, Lysine, Arginine, Pheny
Lualanine, tyrosine, histidine, tryptophan,
Asparagine, glutamine, proline, ornithine, shi
Examples include toluline. As a non-natural amino acid
Is norleucine, γ-methylleucine, tert-leucine
N, norvaline, homoarginine, homoserine, amino
Isobutyric acid, aminoadipic acid (eg, α-aminoadipic acid
Acid), phenylglycine, thienylglycine, cyclo
Hexylglycine, aminobutyric acid, β-alanine, cyclo
Hexylalanine, thienylalanine, naphthylalani
, Adamantylalanine, benzothienylalanine,
Pyridyl alanine, piperidyl alanine, pyrazyl alan
Nin, quinolyl alanine, thiazolyl alanine, isoa
Sparagine, isoglutamine, homocysteine, homov
Phenylalanine, homocitrulline, oxyproline (hi
Droxyproline), diaminopropionic acid, diamino
Butyric acid, aminobenzoic acid, the aforementioned natural amino acids, non-natural amino acids
Examples thereof include N-methylated no acids. These nets
Examples of the substituent that may be substituted on the side chain of the acid residue
Is (1) C_1-6Alkyl group (eg, methyl, ethyl, n
-Propyl, isopropyl, n-butyl, isobutyl,
sec-butyl, tert-butyl, n-pentyl
But preferably C_1-3Alkyl, etc.), (2) cyano
Group, (3) halogen (eg, fluorine, chlorine, bromine, iodine, etc.
DO), (4) Hydroxy-C_1-6Alkyl group (eg, hydr
Roxymethyl, hydroxyethyl, etc.), (5) C_1-6
Alkoxy group (eg, methoxy, ethoxy, n-propoxy)
Ci, isopropoxy, n-butoxy, tert-butoxy
C, etc., preferably C_1-3(Alkoxy etc.), (6) C
_1-6Alkoxy-carbonyl group (eg, methoxycarbo group
Nyl, ethoxycarbonyl, isopropoxy carbonyl
And tert-butoxycarbonyl, etc., preferably C
_1-3(Alkoxy-carbonyl, etc.), (7) C_1-4A
Silyl groups (eg formyl, acetyl, propionyl, buty
Formyl such as ril and C_2-4Alkanoyl, etc.),
(8) Hydroxy group, (9) Formula: —S (O) a—R²¹(In the formula,
a is an integer of 0 to 2, R²¹Is C_1-6Alkyl (specific
Examples include the same as above)))
Groups (eg, methylthio, methanesulfinyl, meta
Sulfonyl, ethylthio, ethanesulfinyl, ethane
Sulfonyl, etc.), (10) benzyloxycarbonyl,
(11) Tosyl group, (12) carbamoyl group, (13) mercapto
Group, (14) amino group, (15) sulfo group, (16) phosphono group, (1
7) phospho group, (18) carboxyl group, (19) tetrazoli
Group, (20) amino-C_1-6Alkyl group (eg, amino
Tyl, aminoethyl, etc.), (21) aminoallyl group, (22)
Thiazolyl group, (23) thienyl group, (24) oxazolyl group,
(25) Furyl group, (26) pyranyl group, (27) pyridyl group, (28)
Pyrazyl group, (29) pyrazinyl group, (30) pyrimidinyl group,
(31) pyridazinyl group, (32) indolyl group, (33) indodi
Nyl group, (34) isoindolyl group, (35) pyrrolyl group, (3
6) imidazolyl group, (37) isothiazolyl group, (38) pyra
Zolyl group, (39) chromenyl group, (40) purinyl group, (41) key
Noridinyl group, (42) quinolyl group, (43) isoquinolyl group,
(44) quinazolinyl group, (45) quinoxalinyl group, (46) syn
Norinyl group, (47) morpholinyl group, (48) benzothienyl
Group, (49) benzofuranyl group, (50) benzimidazole
(51) benzimidazolyl group, (52) C_3-8Cycloa
Rualkyl group, (53) oxo group, (54) from the above (2), (3), (5)
(19), C substituted with the substituent described in (22) to (52)
_1-4Alkyl group, (55) (2), (3), (5) to (1
9), holmi substituted with the substituent described in (22) to (52)
Le, C_2-4C such as alkanoyl_1-4Acyl group, (5
6) Phenyl substituted with the substituent described in the above (1) to (52)
C such as le _6-10Aryl groups (mesityl, tolyl, chi
Silyl, styrenyl, etc.), (57) above (1) to (52)
C such as benzyl substituted with the above substituents_7-15Aralchi
Group (methylbenzyl, methoxybenzyl, etc.), (58)
C_7-15Aralkyl groups (benzyl, phenethyl, benzhi
Drill, naphthylmethyl, etc.), (59) C_6-10Ally
Group (phenyl, naphthyl, indenyl, etc.)
You can

As the neutral substituent, (1) C_1-6Al
Kill group (eg, methyl, ethyl, n-propyl, isopro
Pill, n-butyl, isobutyl, sec-butyl, te
rt-butyl, n-pentyl and the like, preferably C_1-3
Alkyl, etc.), (2) cyano group, (3) halogen (eg, fluorine
Element, chlorine, bromine, iodine, etc.), (4) hydroxy-C
_1-6Alkyl group (eg hydroxymethyl, hydroxy
Ethyl, etc.), (5) hydroxy group, (6) carbamoyl group,
(7) Mercapto group, (8) Formula: —S (O) a—R²¹(Each in the formula
The symbol has the same meaning as defined above, (9) C_6-10Ally
Group (eg phenyl, naphthyl, indenyl, chromenyl)
Etc.), (10) thienyl group, (11) oxazolyl group, (12) phenyl group
Ryl group, (13) indolyl group, (14) indolizinyl group, (1
5) Isoindolyl group, (16) C_3-8Cycloalkyl group,
(17) an oxo group, (18) the above (2), (3), (5) to (16)
C substituted with a substituent_1-6Alkyl, (19) Above (1)
Phenyl and naphthyl substituted with the substituents described in (16)
Such as C_6-10Aryl group (mesityl, tolyl, xy
Ril, styrenyl, etc.), (20) As described in (1) to (16) above
C such as benzyl substituted with the substituents of_7-15Aral
Kill group (methylbenzyl, methoxybenzyl, etc.)
Can be given. The acidic substituents are carbon
C substituted with a xyl group, a sulfo group, a tetrazolyl group, etc.
_1-4C such as alkyl group, phenyl and naphthyl
_6-10C such as aryl group or benzyl_7-15
Examples thereof include aralkyl group and carboxyl group. base
Examples of the volatile substituent include (1) amino-C_1-6Alkyl group
(Aminomethyl, aminoethyl, etc.), (2) aminoant
Group, (3) pyridyl group, (4) pyrazyl group, (5) pyrazinyl
Group, (6) pyridazinyl group, (7) imidazolyl group, (8) pyridine group
Razolyl group, (9) pyrazolyl group, (10) morpholinyl group,
(11) an amino group, (12) a substituent according to the above (3) to (10)
The substituted C_1-4Alkyl group, (13) As described in (1) to (11) above.
C such as benzyl substituted with the above substituents_7-15Ara
A rutile group, (14) substituted with the substituent described in (1) to (11) above
C such as phenyl and naphthyl_6-10Aryl group
And so on. In the present specification, an acidic amino acid and
Then, specifically, for example, in the side chain carboxyl
Having acidic groups such as groups, sulfo groups, tetrazolyl groups
Amino acids are included. As a specific example, glutami
Acid, pyroglutamic acid, aspartic acid, cysteine
Acid, homocysteic acid, 3- (5-tetrazolyl) ara
Nin, 2-amino-4- (5-tetrazolyl) butyric acid, etc.
Can be given. In the present specification, a basic amino acid
For example, histidine, arginine, ortini
, Lysine, diaminopropionic acid, diaminobutyric acid,
Examples include homoarginine. Side chain substituted salt
Specific examples of the basic amino acid include N,^α−
Acetyl Arginine, N^ε-Tosylarginine, N^ε
-Acetyllysine, N^ε-Methyllysine, N^ε-Tosyl
Examples include lysine. As used herein, neutral ami
Specific examples of the acid include, for example, alanine and varieties.
, Norvaline, leucine, isoleucine, alloisoro
Isine, norleucine, tertiary leucine, gamma
Methylleucine, proline, phenylglycine, phenyl
Rualanine, glutamine, asparagine, serine, thread
Onine, glycine, cysteine, methionine, trypto
Fan, oxyproline (hydroxyproline), shiku
Examples include amino acids such as lohexylalanine. ~ side
As the neutral amino acid whose chain has been replaced, specifically,
Examples include biphenylalanine.

In the present specification, examples of the amino acid residue having an aromatic side chain include, for example, tryptophan, phenylalanine, tyrosine, 1-naphthylalanine, 2-naphthylalanine, 2-thienylalanine, Examples thereof include histidine, pyridylalanine (2-pyridylalanine), O-methyltyrosine and the like. Specific examples of the amino acid residue having an aromatic side chain in which the side chain is substituted include, for example, 3-iodotyrosine and p-
Examples thereof include phosphonomethylphenylalanine and O-phosphotyrosine. In the present specification, examples of the amino acid residue having a hydroxy group in the side chain include serine, threonine, tyrosine, oxyproline (hydroxyproline) and the like. In the present specification, peptides and peptide chains have N at the left end according to the convention of peptide notation.
The terminal (amino terminal) and the right terminal are C terminals (carboxyl terminal). Peptides and peptide chains of the invention C-terminus is usually a carboxyl group (-COOH)) or carboxylate (-COO ^- is a) be C-terminal amide (-CONH ₂₎ or an ester (-COOR) Good. R of the ester is, for example, a C _1-6 alkyl group such as methyl, ethyl, n-propyl, isopropyl or n-butyl,
C _3-8 cycloalkyl group such as cyclopentyl and cyclohexyl, C _6-12 such as phenyl and α-naphthyl
In addition to phenyl-C _1-2 alkyl such as aryl group, benzyl, phenethyl, benzhydryl, etc., or C _7-14 aralkyl group such as α-naphthyl-C _1-2 alkyl such as α-naphthylmethyl, it is also widely used as an oral ester. And a pivaloyloxymethyl group. When the peptide of the present invention has a carboxyl group or carboxylate other than the C-terminal, those in which the group is amidated or esterified are also included in the polypeptide of the present invention. As the ester at this time, for example, the above-mentioned C-terminal ester or the like is used. Moreover, in the peptide or peptide chain of the present invention, the amino group of the amino acid residue at the N-terminal is a substituent (for example, C _2-6 alkanoyl group such as formyl and acetyl, guanidinoacetyl, thienylacrylyl, pyridylacetyl and the like). C _1-8 acyl group, methyl, ethyl, propyl,
Isopropyl, butyl, isobutyl, sec-butyl, ter
C _1-6 alkyl group such as t-butyl, pentyl and hexyl, C _6-10 aryl group such as phenyl and naphthyl, or C _7-16 aralkyl group such as benzyl and phenethyl, tosyl group, benzyloxycarbonyl group,
Formula: —S (O) a—R ²² (in the formula, a represents an integer of 0 to 2, and R ²² represents C _1-6 alkyl (specific examples thereof include the same ones as described above)). Those substituted with a group represented (eg, methylthio, methanesulfinyl, methanesulfonyl, ethylthio, ethanesulfinyl, ethanesulfonyl, etc.), t-butoxycarbonyl group, N-9-fluorenylmethoxycarbonyl group, etc., N-terminal glutamine residue produced by cleavage in vivo is oxidized by pyroglutamine, substituents on side chains of amino acids in the molecule (for example, -OH, -SH, amino group, imidazolyl group, indolyl group, guanidino group) A group such as C is a suitable protecting group (eg, formyl group, acetyl group, etc.)
Protected by a C _1-6 acyl group such as a _1-6 alkanoyl group) or a complex protein such as a so-called glycoprotein having a sugar chain bound thereto.

The modified form (I) of apelin has the formula X ¹ -Ar
^{g-Pro-Arg-X 2} -Ser-His-X 3 -G
It is represented by ly-Pro-X ⁴ -X ⁵ , and is -Arg-Pro.
-Arg-, -Ser-His- and -Gly-Pr
The side chain of each amino acid residue in o- may be substituted, and examples of the substituent include the above-mentioned substituents and the like.
In the present specification, X ¹ represents “a hydrogen atom or an amino acid residue or a peptide chain consisting of 1 to 25 amino acids which may be the same or different and whose side chains may be substituted”. Examples of the substituent in the “1 to 25 amino acids whose side chain may be substituted” include, for example,
Examples thereof include those similar to the above-mentioned “substituents which may be substituted on the side chain of an amino acid residue”. Preferred examples when X ¹ represents “an amino acid residue whose side chain may be substituted” include, for example, optionally substituted pyroglutamic acid and optionally substituted side chain glutamine. And more preferably pyroglutamic acid or glutamine. Examples of the substituent of the amino acid residue in the “side chain optionally substituted amino acid residue”, “optionally substituted pyroglutamic acid”, and “side chain optionally substituted glutamine” include The same as the above "substituent which may be substituted on the side chain of the amino acid residue" and the like can be mentioned. Moreover, a benzyloxycarbonyl group etc. are mentioned as a preferable substituent of "pyroglutamic acid which may be substituted". Specific examples of when X ¹ represents “a peptide chain consisting of 2 to 25 amino acids which may be the same or different and whose side chains may be substituted” include, for example, the formula Y
^1- Y ² (In the formula, Y ¹ is the same or different and represents an amino acid residue or a peptide chain consisting of 1 to 17 amino acids whose side chains may be substituted, and Y ² is the same or different. A peptide represented by (an amino acid residue or a peptide chain consisting of 1 to 8 amino acids whose side chain may be substituted) is shown.

Y above¹And Y^TwoAmino acid residue represented by
Groups or side chain substituents of amino acid residues in the peptide chain
For example, substituting the side chain of the amino acid residue described above
And the like.
Above Y¹As a specific example of, for example, (a) formula A
¹-A^Two-A^Three-A^Four-A⁵-A⁶-A⁷-A⁸-A⁹
-A¹⁰-A ¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵−
A¹⁶-A¹⁷(In the formula, A¹Through A ¹⁷Are the same
One or different amino acids whose side chains may be substituted
Residue), (b) Formula A^Two-A^Three-A^Four-A⁵-A⁶
-A⁷-A⁸-A⁹-A¹⁰-A¹¹-A¹²-A^Thirteen
-A¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol is
The same meaning as above), (c) Formula A^Three-A^Four-A⁵-A⁶
-A⁷-A⁸-A⁹-A¹⁰-A¹¹-A¹²-A^Thirteen
-A¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol is
The same meaning as above), (d) Formula A^Four-A⁵-A⁶-A⁷
-A⁸-A⁹-A¹⁰-A¹¹-A¹²-A^{1 Three}-A
¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol is as described above.
(Same meaning), (e) Formula A⁵-A⁶-A⁷-A⁸-A
⁹-A¹⁰-A¹¹-A¹²-A^Thirteen-A ¹⁴-A¹⁵
-A¹⁶-A¹⁷(In the formula, each symbol has the same meaning as above.
), (F) Formula A⁶-A⁷-A⁸-A⁹-A¹⁰-A
¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as described above), (g) formula
A⁷-A⁸-A⁹-A¹⁰-A¹¹-A¹²-A^Thirteen−
A¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol is as described above.
And (h) formula A⁸-A⁹-A¹⁰-A
¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵-A^{1 6}-A
¹⁷(Wherein each symbol has the same meaning as described above), (i) formula
A⁹-A¹⁰-A¹¹-A¹²-A^Thirteen-A¹⁴-A
¹⁵-A¹⁶-A ¹⁷(In the formula, each symbol has the same meaning as above.
Shown), (j) Formula A¹⁰-A¹¹-A¹²-A^Thirteen-A
¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol is as described above.
Have the same meaning), (k) Formula A¹¹-A¹²-A^Thirteen-A
¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol is as described above.
Have the same meaning), (l) Formula A¹²-A^Thirteen-A¹⁴-A
¹⁵-A¹⁶-A¹⁷(In the formula, each symbol has the same meaning as above.
Shown), (m) Formula A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as described above), (n) formula
A¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol is as described above.
The same meaning as), (0) Formula A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as above), (p) formula
A¹⁶-A¹⁷(In the formula, each symbol has the same meaning as above)
Or (q) A¹⁷(A¹⁷Represents the same meaning as above)
Amino acid residues or peptide chains
It

The above-mentioned A ¹ represents an amino acid residue whose side chain may be substituted, preferably an amino acid residue having an aromatic side chain, and more preferably L-having an aromatic side chain. Amino acid residues, more preferably L-tyrosine and the like are shown. The above A ² and A ³ are the same or different and each represents an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably , L-neutral amino acid residue whose side chain may be substituted, more preferably, A ² is L-leucine and the like, and A ³ is L-valine and the like. The above A ⁴ represents an amino acid residue whose side chain may be substituted, preferably a neutral or basic amino acid residue whose side chain may be substituted, more preferably a side chain substituted. Optionally L-neutral or L-basic amino acid residue, more preferably L-lysine or N ^ε -acetyllysine. The above A ⁵ represents an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably L-which may be substituted. Proline, more preferably L-proline and the like are shown. The above A ⁶ and A ⁹ are the same or different and each represent an amino acid residue whose side chain may be substituted, preferably a basic amino acid residue whose side chain may be substituted, and more preferably , L-basic amino acid residue whose side chain may be substituted, more preferably L-arginine. The above A ⁷ and A ¹⁰ are the same or different and each represent an amino acid residue whose side chain may be substituted, but preferably, an amino acid residue having a hydroxy group in the side chain or a side chain is substituted. It also indicates a neutral amino acid residue. As A ⁷ , glycine which may be substituted, particularly glycine, is preferably used. The above A ⁸ represents an amino acid residue whose side chain may be substituted, preferably an amino acid residue having L-proline or a hydroxy group in its side chain, more preferably L-serine, L
-Indicates proline or oxyproline (hydroxyproline) and the like. The above A ¹⁰ is preferably an amino acid residue having a hydroxy group in its side chain or a neutral amino acid residue whose side chain may be substituted, more preferably serine, threonine, asparagine and the like. A ¹¹ above
To A ¹⁴ are the same or different and each represent an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably A ¹¹ Are glycine, A ¹² is L-proline, A ¹³ is glycine, and A ¹⁴ is L-alanine, L-proline and the like. The above A ¹⁵ represents an amino acid residue whose side chain may be substituted, preferably an amino acid residue having an aromatic side chain, more preferably an L-amino acid having an aromatic side chain. Residues, more preferably L-tryptophan and the like are shown. The above A ¹⁶ represents an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably a neutral L- having a carbamoyl group. Amino acid residue, more preferably L-
Indicates glutamine. Examples of the neutral L-amino acid residue having a carbamoyl group include L-glutamine and L-asparagine. A ¹⁷ represents an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably glycine and the like.

Y above^TwoFor example,
If  Formula B¹-B^Two-B^Three-B^Four-B⁵-B⁶-B⁷-B
⁸(In the formula, B¹Through B ⁸Are the same or different
Represents an amino acid residue whose side chain may be substituted),  Formula B^Two-B^Three-B^Four-B⁵-B⁶-B⁷-B⁸(formula
, Each symbol has the same meaning as above),  Formula B^Three-B^Four-B⁵-B⁶-B⁷-B⁸(Each in the formula
The symbols have the same meaning as above),  Formula B^Four-B⁵-B⁶-B⁷-B⁸(In the formula, each symbol is
(Same meaning as above),  Formula B⁵-B⁶-B⁷-B⁸(In the formula, each symbol is as above.
Have the same meaning),  Formula B⁶-B⁷-B⁸(In the formula, each symbol has the same meaning as above.
),  Formula B⁷-B⁸(In the formula, each symbol has the same meaning as above.
Or)  Formula B⁸(B⁸Represents the same meaning as above)
Examples include mino acid residues and peptide chains. B above¹Is an amino acid residue whose side chain may be substituted
Although shown, preferably, the side chain may be substituted neutral
An amino acid residue, more preferably a side chain substituted
A neutral L-amino acid residue, more preferably a substitution
Optionally glycine, most preferably glycine
Etc. B above^TwoThrough B^FourAre the same or
Differently, the amino acid residue whose side chain may be substituted
Although shown, preferably, a base whose side chain may be substituted
Amino acid residues, more preferably side chains substituted
Optionally a basic L-amino acid residue, more preferably
B^TwoAs L-arginine, B^ThreeAs L-Argi
Nin, B^FourShows L-lysine and the like. B above⁵
Represents an amino acid residue whose side chain may be substituted,
Preferably, an amino acid residue having an aromatic side chain,
More preferably, the L-amino acid residue having an aromatic side chain
Group, more preferably L-phenylalanine
You B above⁶And B⁷Are the same or different
Indicates an amino acid residue whose side chain may be substituted
However, preferably, a basic group in which a side chain may be substituted is used.
Minoic acid residue, more preferably even if the side chain is substituted
A good basic L-amino acid residue, more preferably L-
Indicates arginine, etc. B above⁸Has side chains replaced
Amino acid residue which may be present, but preferably the side chain is
Optionally substituted glutamine, more preferably L-
Indicates glutamine, etc.

Y¹And Y^TwoAs a combination of
When expressed by (a) expression + above expression
(Ie X¹But A ¹-A^Two-A^Three-A^Four-A⁵-A⁶−
A⁷-A⁸-A⁹-A¹⁰-A¹¹-A^{1 Two}-A^Thirteen−
A¹⁴-A¹⁵-A¹⁶-A¹⁷-B¹-B^Two-B^Three−
B^Four-B⁵-B⁶-B⁷-B⁸When represented by:
It is omitted in the explanation of the combination), and is shown in (a) above.
When expressed by the above formula + the above formula, the above
(a) + the above expression
If it is expressed as (a) above + the above expression
If the above is true, the formula represented by (a) above + the formula represented by above
When expressed by, the expression expressed in (a) above + expressed in the above
When expressed by the formula, the formula expressed by (a) above + the above
When expressed by the formula, the formula expressed by (a) above +
When represented by the formula represented above, represented by (b) above
When expressed by the formula + the above formula, in (b) above
When expressed by the expression + the above expression, the above
(b) + the above expression
In this case, the formula expressed by (b) above + the formula expressed by above
If the above is true, the formula expressed in (b) above + the formula expressed above
When expressed by, the formula expressed by (b) above + expressed by the above
When expressed by the formula, the formula expressed by (b) above + the above
When expressed by the formula, the formula expressed by (b) above +
When represented by the formula represented above, represented by the above (c)
In case of the formula + the formula above, in (c) above
When expressed by the expression + the above expression, the above
(c) + (expression above)
In this case, it is represented by the formula (c) above + the formula above
If the above is true, the formula represented by (c) above + the formula represented by above
When expressed by, the formula expressed by (c) above + expressed by the above
When expressed by the formula, the formula expressed by (c) above + the above
When expressed by the formula, the formula expressed by (c) above +
When represented by the formula represented above, it is represented by (d) above.
In the case of the formula + the formula shown above, in (d) above
When expressed by the expression + the above expression, the above
(d) + the above expression
In this case, it is represented by the formula represented by (d) above + the formula represented by above
In case of the above, the formula represented by the above (d) + the formula represented by the above
When expressed by, the formula expressed by (d) above + expressed by the above
When expressed by the formula, the formula expressed by (d) above + the above
When expressed by the formula, the formula expressed by (d) above +
When represented by the formula represented above, it is represented by (e) above.
In case of the formula + the formula shown above, in (e) above
When expressed by the expression + the above expression, the above
(e) + the above expression
In this case, the formula expressed by (e) above + the formula expressed by above
If the above is true, the formula expressed in (e) above + the formula expressed above
When expressed by, the expression expressed by (e) above + expressed by the above
When expressed by the formula, the formula expressed by (e) above + the above
When expressed by the formula, the formula expressed by (e) above +
When represented by the formula represented above, it is represented by (f) above.
In the case of being expressed by the formula + the above formula, in (f) above
When expressed by the expression + the above expression, the above
(f) + the above expression
In this case, it is expressed by the formula shown in (f) above + the formula shown above
In case of the above, the formula represented by the above (f) + the formula represented by the above
When expressed by, the formula expressed by (f) above + expressed by
When expressed by the formula, the formula expressed by (f) above + the above
When expressed by the formula, the formula expressed by (f) above +
When represented by the formula represented above, represented by (g) above
Formula + the above formula, in (g) above
When expressed by the expression + the above expression, the above
(g) + the above expression
If it is expressed by the formula (g) above + the formula above
If the above is true, the formula represented by the above (g) + the formula represented by the above
When expressed by, the formula expressed by (g) above + expressed by
When expressed by the formula, the formula expressed by (g) above + the above
When expressed by the formula, the formula expressed by (g) above +
When represented by the formula represented above, it is represented by (h) above.
In the case of being expressed by the formula + the above formula, in (h) above
When expressed by the expression + the above expression, the above
(h) + the above expression
In this case, it is expressed by the formula represented by (h) + the formula represented by the above.
In case of the above, the formula represented by the above (h) + the formula represented by the above
When expressed by, the formula expressed by (h) above + expressed by the above
When expressed by the formula, the formula expressed by (h) above + the above
When expressed by the formula, the formula expressed by (h) above +
When represented by the formula represented above, represented by (i) above
In the case of the above formula + the above formula, in (i) above
When expressed by the expression + the above expression, the above
(i) + the above expression
In this case, it is represented by the formula (i) above + the formula above
If the above is true, the formula represented by (i) above + the formula represented by above
When expressed by, the expression expressed by (i) above + expressed by the above
When expressed by the formula, the formula expressed by (i) above + the above
When expressed by the formula, the formula expressed by (i) above +
When represented by the above formula, it is represented by (j) above.
Formula + the above formula, in (j) above
When expressed by the expression + the above expression, the above
(j) + (expression above)
If it is expressed as (j) above + the above expression
If the above is true, the formula expressed in (j) above + the formula expressed above
When expressed by, the expression expressed by (j) above + expressed by the above
When expressed by the formula, the formula expressed by (j) above + the above
When expressed by the formula, the formula expressed by (j) above +
When represented by the formula represented above, it is represented by (k) above.
In the case of being expressed by the formula + the above formula, in (k) above
When expressed by the expression + the above expression, the above
(k) + the above expression
If it is expressed by the above formula (k) + the above formula,
In case of the above, the formula represented by the above (k) + the formula represented by the above
When expressed by, the expression expressed by (k) above + expressed by the above
When expressed by the formula, the formula expressed by (k) above + the above
When expressed by the formula, the formula expressed by (k) above +
When represented by the formula represented above, it is represented by (l) above.
Formula + the above formula, in (l) above
When expressed by the expression + the above expression, the above
(l) + the above expression
The formula expressed by (l) above + the formula expressed by above
If the above is true, the formula represented by (l) above + the formula represented above
When expressed by, the formula expressed by (l) above + the above expressed
When expressed by the formula, the formula expressed by (l) above + the above
When expressed by the formula, the formula expressed by (l) above +
When represented by the formula represented above, it is represented by (m) above.
In the case of being expressed by the formula + the above formula, in (m) above
When expressed by the expression + the above expression, the above
(m) + the above expression
If it is expressed by the formula (m) above + the formula above
In case of the above, the formula represented by the above (m) + the formula represented by the above
When expressed by, the formula expressed by (m) above + expressed by the above
When expressed by the formula, the formula expressed by (m) above + the above
When expressed by the formula, the formula expressed by (m) above +
When represented by the formula represented above, represented by (n) above
When expressed by the formula + the formula above, in (n) above
When expressed by the expression + the above expression, the above
(n) + the above expression
In this case, it is represented by the formula represented by (n) above + the formula represented by above
If the above is true, the formula represented by (n) above + the formula represented above
Is expressed by the above formula (n) + the above
When expressed by the formula, the formula expressed by (n) above + the above
When expressed by the formula, the formula expressed by (n) above +
When represented by the formula represented above, it is represented by (o) above.
In case of the formula + the formula shown above, the above (o)
When expressed by the expression + the above expression, the above
(o) + the above expression
If it is expressed as (o) above + the above expression
If the above is true, the formula represented by (o) above + the formula represented by above
When expressed by, the expression expressed by (o) above + expressed by the above
When expressed by the formula, the formula expressed by (o) above + the above
When expressed by the formula, the formula expressed by (o) above +
When represented by the formula represented above, it is represented by (p) above.
In the case of the formula + the formula shown above, in (p) above
When expressed by the expression + the above expression, the above
(p) + the above expression
In this case, it is expressed by the formula (p) above + the formula above
In case of the above, the formula represented by the above (p) + the formula represented by the above
When expressed by, the expression expressed by (p) above + expressed by the above
When expressed by the formula, the formula expressed by (p) above + the above
When expressed by the formula, the formula expressed by (p) above +
When expressed by the above formula, it is expressed by (q) above.
In the case of the formula + the formula shown above, the above (q)
When expressed by the expression + the above expression, the above
(q) + the above expression
If it is expressed as (q) above + (expression above)
In case of the above, the formula represented by the above (q) + the formula represented by the above
When expressed by, the expression expressed by (q) above + expressed by the above
When expressed by the formula, the formula expressed by (q) above + the above
When represented by the formula represented by, and represented by (q) above
There are cases where it is expressed by the formula + the formula shown above.
However, above all, it is expressed by the formula (a) above + the above
When expressed by the formula, the formula expressed by (b) above +
When represented by the formula represented, the formula represented by (c) above +
When represented by the formula described below, it is represented by the above (d)
When expressed by the formula + the formula above, the formula (e) above
When expressed by the above formula + the above formula, the above
(f) + the above expression
If it is expressed by the formula (g) above + the formula above
In case of the above, the formula represented by the above (h) + the formula represented by the above
When expressed by, the expression expressed by (i) above + expressed by the above
When expressed by the formula, the formula expressed by (j) above + the above
When expressed by the formula, the formula expressed by (k) above +
When represented by the formula represented above, it is represented by (l) above.
In the case of being expressed by the formula + the above formula, in (m) above
When expressed by the expression + the above expression, the above
(n) + the above expression
If it is expressed as (o) above + the above expression
If the above is true, the formula expressed in (p) above + the formula expressed above
And the formula (q) above + the above
The case of being represented by the formula

Above all, the formula represented by the above (a) + the above
When represented by the formula represented by and represented by (b) above
The case of being represented by the formula + the formula shown above is more preferable.
Take as an example. Formula (a) above + above
When represented by the formula represented and the formula represented by (b) above
+ Further preferred ingredients when represented by the formulas described above
A body example is shown below. Formula shown in (b) above + Table shown above
Is represented by the formula¹Is the expression A ^Two-A^Three−
A^Four-A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰-A¹¹
-A¹²-A ^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷−
B¹-B^Two-B^Three-B^Four-B⁵-B⁶-B⁷-B⁸(formula
Medium, A²Through A¹⁷And B¹Through B⁸Is the same as above
In this case,
As a preferred specific example of^TwoAnd A^ThreeRespectively
L-, which may be the same or different and whose side chain may be substituted.
Neutral amino acid residue, preferably A^TwoAs L-Royce
A, etc.^ThreeIs L-valine or the like, and A^FourIs on the side
L-neutral or L-basic amide, whose chain may be substituted
Mino acid residue, preferably L-glutamine, L-lysine or
Or N^ε-Acetyllysine, etc., A⁵Is the side chain
Optionally substituted L-proline, preferably L-proline
Such as phosphorus, A⁶And A⁹Are the same or
Differently, the L-basic amino group may have a side chain substituted.
Acid residue, preferably L-arginine and the like,⁷
Optionally substituted glycine, preferably glycine
And so on, A⁸Is an L-proline or hydroxy group
An amino acid residue having a side chain, preferably L-serine,
L-proline or oxyproline (hydroxyproline
, Etc., and A¹⁰Has a hydroxy group in the side chain
Neutral amino acid residue or side chain may be substituted
Amino acid residue, preferably L-serine, L-threoni
A, L-asparagine, etc.¹¹Is glycine,
A¹²Is L-proline, A^ThirteenIs glycine, A¹⁴Is L-
Such as alanine or L-proline, A¹⁵Is fragrance
L-amino acid residue having an aromatic side chain, preferably L-
Tryptophan, etc., A¹⁶Is a carbamoyl group
Having a neutral L-amino acid residue, preferably L-glutamine
And A¹⁷Is a neutral amino acid residue, preferably
Shin, etc.,

B ¹ is a neutral L in which the side chain may be substituted.
-An amino acid residue, preferably glycine which may be substituted, more preferably glycine and the like, in which B ² to B ⁴ are the same or different from each other, and a basic L- whose side chain may be substituted- An amino acid residue, preferably
B ² is L-arginine, B ³ is L-arginine, B ⁴ is L-lysine, etc., and B ⁵ is an L-amino acid residue having an aromatic side chain, preferably,
L-phenylalanine, etc., B ⁶ and B ⁷ are the same or different, and a basic L-amino acid residue whose side chain may be substituted, preferably L-arginine, etc., and B ⁸ is Examples thereof include the case where the chain is glutamine which may be substituted, preferably L-glutamine.

Formula represented by the above (a) + represented by the above
When expressed by a formula, X¹Is the expression A ¹-A^Two-A^Three−
A^Four-A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰-A¹¹
-A^{1 Two}-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷−
B¹-B^Two-B^Three-B^Four-B⁵-B⁶-B⁷-B⁸(formula
Medium, A¹Through A¹⁷And B¹Through B⁸Is the same as above
In this case,
As a preferred specific example of¹Has aromatic side chains
L-amino acid residue, more preferably L-tyrosine
And A^TwoAnd A^ThreeAre the same or different
And an L-neutral amino acid residue whose side chain may be substituted
A group, preferably A^TwoAs L-leucine, A^ThreeWhen
Is L-valine, and A^FourHas side chains replaced
L-neutral or L-basic amino acid residue which may be
More preferably, L-glutamine, L-α aminoadipic acid, L-
Lysine or N^ε-Acetyllysine, etc., A⁵But
Optionally substituted L-proline, preferably L-proline
Such as Lorin, A⁶And A⁹Are the same
Are different from each other, and L-basic acid optionally substituted in the side chain.
A mino acid residue, preferably L-arginine and the like, A
⁷Optionally substituted glycine, preferably glycine
A, etc., and⁸Is L-proline or hydroxy group
An amino acid residue having a side chain, preferably L-seri
, L-proline or oxyproline (hydroxyproline
Lorin) etc., and A¹⁰Has a hydroxy group in the side chain
The amino acid residue or side chain to
Amino acid residue, preferably L-serine, L-threoni
A, L-asparagine, etc.¹¹Is glycine,
A¹²Is L-proline, A^ThirteenIs glycine, A¹⁴Is L-
Such as alanine or L-proline, A¹⁵Is fragrance
L-amino acid residue having an aromatic side chain, preferably L-
Tryptophan, etc., A¹⁶Is a carbamoyl group
Having a neutral L-amino acid residue, preferably L-glutamine
And A¹⁷Is a neutral amino acid residue, preferably
Shin, etc.,

B ¹ is a neutral L in which the side chain may be substituted.
-An amino acid residue, preferably glycine which may be substituted, more preferably glycine and the like, in which B ² to B ⁴ are the same or different from each other, and a basic L- whose side chain may be substituted- An amino acid residue, preferably
B ² is L-arginine, B ³ is L-arginine, B ⁴ is L-lysine, etc., and B ⁵ is an L-amino acid residue having an aromatic side chain, preferably,
L-phenylalanine, etc., B ⁶ and B ⁷ are the same or different, and a basic L-amino acid residue whose side chain may be substituted, preferably L-arginine, etc., and B ⁸ is Examples thereof include the case where the chain is glutamine which may be substituted, and preferably L-glutamine L-pyroglutamic acid.

As more specific and preferable examples of X ¹ , (1) hydrogen atom, (2) Leu-Val-Gln-Pro-Arg-Gly-Ser-
Arg-Asn-Gly-Pro-Gly-Pro-Trp-Gln-Gly-Gly-Arg-Arg-Ly
s-Phe-Arg-Arg-Gln, (3) pGlu, (4) Leu-Val-Adi (NH ₂ )-
Pro-Arg-Gly-Ser-Arg-Asn-Gly-Pro-Gly-Pro-Trp-Gln-Gl
y-Gly-Arg-Arg-Lys-Phe-Arg-Arg-Gln, (5) Leu-Val-Ly
s (Ac) -Pro-Arg-Thr-Ser-Arg-Thr-Gly-Pro-Gly-Ala-Trp-
Gln-Gly-Gly-Arg-Arg-Lys-Phe-Arg-Arg-Gln, (6) Tyr-
Leu-Val-Lys-Pro-Arg-Thr-Ser-Arg-Thr-Gly-Pro-Gly-Al
a-Trp-Gln-Gly-Gly-Arg-Arg-Lys-Phe-Arg-Arg-Gln, and (7) Z-pGlu.

In the present specification, X ² represents a neutral amino acid residue in which the side chain may be substituted, but preferably L-leucine in which the side chain may be substituted, or the side chain is substituted. Optionally L-norleucine, more preferably L
-Leucine, L-norleucine and the like.

In the present specification, X ³ represents a neutral amino acid residue whose side chain may be substituted or a basic amino acid residue whose side chain may be substituted. Examples of the substituent on the side chain of the basic amino acid residue include C _1-4 acyl group, tosyl group, C _1-6 alkyl group and the like.
Examples of the C _1-4 acyl group include formyl such as formyl, acetyl, propionyl and butyryl, and C.
Examples include _2-4 alkanoyl. Examples of the C _1-6 alkyl group include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl and the like. Preferred examples of X ³ include L-lysine whose side chain may be substituted, L-norleucine whose side chain may be substituted, and L-arginine whose side chain may be substituted. More preferably, the side chain is a C _1-4 acyl group (eg, formyl such as formyl, acetyl, propionyl, butyryl and C _2-4 alkanoyl), C
_1-6 alkyl group (eg, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert.
-Butyl, pentyl, hexyl, etc.) or L-lysine optionally substituted with a tosyl group, L-norleucine or L-arginine, and the like, more preferably L-lysine, L-norleucine, L-arginine,
N ^epsilon - acetyllysine, N ^epsilon - methyllysine, N ^epsilon -
Examples thereof include tosyl lysine and N ^ε -tosyl arginine.

In the present specification, X ⁴ represents a neutral or aromatic amino acid residue which may have a bond or a side chain substituted. As X ⁴ , a bond, L-norleucine whose side chain may be substituted, L-methionine whose side chain may be substituted, and L-methionine sulfoxide whose side chain may be substituted. Or L-alanine which may have a side chain substituted, and the like, more preferably a bond, L-norleucine or L-methionine,
Examples thereof include L-methionine sulfoxide and L-cyclohexylalanine.

In the present specification, X ⁵ is an amino acid residue whose side chain may be substituted or an amino acid derivative in which its C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, a hydroxyl group or a side chain is substituted. A dipeptide chain in which an amino acid residue which may be substituted and an amino acid residue whose side chain is substituted or a C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group is shown as a peptide derivative. X ⁵ is preferably a neutral amino acid residue whose side chain may be substituted or an amino acid derivative in which its C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, or a hydroxyl group or side chain may be substituted. A dipeptide chain comprising a good neutral amino acid residue and an amino acid residue having an aromatic side chain which may be substituted in the side chain or its C-terminal carboxyl group is reduced to a hydroxylmethyl group or a formyl group. And peptide derivatives. More preferably, X ⁵ is an amino acid derivative in which the side chain may be substituted, L-proline or its C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, and the side chain may be substituted 4- Chlorophenylalanine or an amino acid derivative whose C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, 2-naphthylalanine whose side chain may be substituted or its C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group Amino acid derivative, a side chain-substituted cyclohexylalanine or an amino acid derivative whose C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, a hydroxyl group or an optionally substituted L-proline and (a ) L- whose side chain may be substituted Phenylalanine, (b) side chain optionally substituted L-tyrosine, (c) side chain optionally substituted L-2-thienylalanine, (d) side chain optionally substituted L -Phenylglycine or (e) a dipeptide chain having L-2-pyridylalanine optionally substituted on its side chain, or a peptide derivative in which its C-terminal carboxyl group is reduced to a hydroxylmethyl group or a formyl group, etc. Can be given. More preferably, X ⁵ is (1) L
-Proline or an amino acid derivative whose C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, (2) 4-chlorophenylalanine or an amino acid derivative whose C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, ( 3) Amino acid derivative in which 2-naphthylalanine or its C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, (4)
Amino acid derivative in which cyclohexylalanine or its C-terminal carboxyl group is reduced to a hydroxymethyl group or formyl group, (5) hydroxyl group, (6) dipeptide chain formed by binding L-proline and L-phenylalanine or its C-terminal carboxyl group A peptide derivative in which is reduced to a hydroxylmethyl group or formyl group, (7) A peptide derivative in which L-proline and L-tyrosine are bound or a C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group , (8) L-proline and L-2-thienylalanine-bonded dipeptide chain or its C-terminal carboxyl group reduced to a hydroxylmethyl group or a formyl group, (9) L-proline and L- A dipeptide chain or its bound to phenylglycine A peptide derivative having a C-terminal carboxyl group reduced to a hydroxylmethyl group or a formyl group, (10) a dipeptide chain formed by binding L-proline and 4-chlorophenylalanine or its C-terminal carboxyl group to a hydroxylmethyl group or a formyl group. Reduced peptide derivative, (11) L-proline and 2
A dipeptide chain having naphthylalanine bound thereto or a peptide derivative having a C-terminal carboxyl group reduced to a hydroxylmethyl group or a formyl group, (12) a dipeptide chain having L-proline bound to 3-iodotyrosine or A peptide derivative in which the C-terminal carboxyl group is reduced to a hydroxylmethyl group or a formyl group, (1
3) A peptide derivative in which L-proline and O-methyltyrosine are bound or a C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group, or (14) L-proline and L-2-pyridyl Examples thereof include a dipeptide chain formed by binding alanine or a peptide derivative in which the C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group.

As "-X ⁴ -X ⁵ ", particularly preferably (1) -Nle-Pro-Phe, (2) -Nle-Pro-Tyr, (3) -Nle-P.
ro, (4) -Nle, (5) -Met-Pro-Phe, (6) -Nle-Pro-Thi,
(7) -Nle-Pro-Phg, (8) -Nle-Pro-Pya (2), (9) -Met
(O), (10) -Met-Phe (Cl), (11) -Met-Pro-Phe (Cl), (1
2) -Met-Pro-Nal (2), (13) -Met-Nal (2), (14) -Met-Ch
a, (15) -Cha-Pro-Phe, (16) -Cha, (17) -Met-Pro-Tyr
(I) and (18) -Met-Pro-Tyr (Me).

Specific examples of the modified apelin (I) include (1) Leu-Val-Gln-Pro-Arg-Gly-Ser-Arg-As.
n-Gly-Pro-Gly-Pro-Trp-Gln-Gly-Gly-Arg-Arg-Lys-Phe-
Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nl
e-Pro-Phe (SEQ ID NO: 13), (2) Leu-Val-Gln-Pro
-Arg-Gly-Ser-Arg-Asn-Gly-Pro-Gly-Pro-Trp-Gln-Gly-G
ly-Arg-Arg-Lys-Phe-Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser
-His-Lys-Gly-Pro-Nle-Pro-Tyr (SEQ ID NO: 14),
(3) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle-P
ro-Phe (SEQ ID NO: 15), (4) pGlu-Arg-Pro-Arg-L
eu-Ser-His-Lys-Gly-Pro-Nle-Pro-Tyr (SEQ ID NO: 1
6), (5) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro
-Nle-Pro (SEQ ID NO: 17), (6) pGlu-Arg-Pro-Arg
-Leu-Ser-His-Lys-Gly-Pro-Nle (SEQ ID NO: 18),
(7) Ac-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle-Pro
-Tyr (SEQ ID NO: 19), (8) Ac-Arg-Pro-Arg-Leu-S
er-His-Lys-Gly-Pro-Nle-Pro (SEQ ID NO: 20),
(9) Ac-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle
(SEQ ID NO: 21), (10) pGlu-Arg-Pro-Arg-Leu-Ser-
His-Lys (Ac) -Gly-Pro-Met-Pro-Phe (SEQ ID NO: 2
2), (11) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys (Me) -Gl
y-Pro-Met-Pro-Phe (SEQ ID NO: 23), (12) pGlu-A
rg-Pro-Arg-Leu-Ser-His-Lys (Ac) -Gly-Pro-Nle-Pro-Phe
(SEQ ID NO: 24), (13) pGlu-Arg-Pro-Arg-Leu-Se
r-His-Lys (Me) -Gly-Pro-Nle-Pro-Phe (SEQ ID NO: 2
5), (14) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys (Tos) -G
ly-Pro-Nle-Pro-Phe (SEQ ID NO: 26), (15) pGlu-
Arg-Pro-Arg-Leu-Ser-His-Arg (Tos) -Gly-Pro-Nle-Pro-P
he (SEQ ID NO: 27), (16) pGlu-Arg-Pro-Arg-Nle-
Ser-His-Lys-Gly-Pro-Nle-Pro-Phe (SEQ ID NO: 2
8), (17) pGlu-Arg-Pro-Arg-Nle-Ser-His-Lys-Gly-Pr
o-Nle-Pro-Tyr (SEQ ID NO: 29), (18) pGlu-Arg-P
ro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle-Pro-Thi (SEQ ID NO: 30), (19) pGlu-Arg-Pro-Arg-Leu-Ser-His-Ly
s-Gly-Pro-Nle-Pro-Phg (SEQ ID NO: 31), (20) pG
lu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle-Pro-Pya
(2) (SEQ ID NO: 32), (21) Arg-Pro-Arg-Leu-Ser
-His-Lys-Gly-Pro-Nle-Pro-Tyr (SEQ ID NO: 33),
(22) Leu-Val-Adi (NH ₂ ) -Pro-Arg-Gly-Ser-Arg-Asn-Gly-
Pro-Gly-Pro-Trp-Gln-Gly-Gly-Arg-Arg-Lys-Phe-Arg-Ar
g-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle-Pro-
Phe (SEQ ID NO: 34), (23) Leu-Val-Lys (Ac) -Pro-
Arg-Thr-Ser-Arg-Thr-Gly-Pro-Gly-Ala-Trp-Gln-Gly-Gl
y-Arg-Arg-Lys-Phe-Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-
His-Lys-Gly-Pro-Nle-Pro-Tyr (SEQ ID NO: 35),
(24) Tyr-Leu-Val-Lys-Pro-Arg-Thr-Ser-Arg-Thr-Gly-P
ro-Gly-Ala-Trp-Gln-Gly-Gly-Arg-Arg-Lys-Phe-Arg-Arg
-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle-Pro-P
he (SEQ ID NO: 36), (25) Z-pGlu-Arg-Pro-Arg-Le
u-Ser-His-Lys (Ac) -Gly-Pro-Nle-Pro-Phe (SEQ ID NO: 37), (26) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-H
is-Lys-Gly-Pro-Met (O) (SEQ ID NO: 38), (27) Ar
g-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle-
Pro-Tyr (SEQ ID NO: 39), (28) pGlu-Arg-Pro-Arg
-Leu-Ser-His-Lys-Gly-Pro-Met-Phe (Cl) (SEQ ID NO:
40), (29) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-
Pro-Met-Pro-Phe (Cl) (SEQ ID NO: 41), (30) Arg-
Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Nal (2)
(SEQ ID NO: 42), (31) Arg-Pro-Arg-Leu-Ser-His-L
ys-Gly-Pro-Met-Nal (2) (SEQ ID NO: 43), (32) Ar
g-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe (Cl)
(SEQ ID NO: 44), (33) Arg-Pro-Arg-Leu-Ser-His
-Lys-Gly-Pro-Met-Phe (Cl) (SEQ ID NO: 45), (34)
Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Cha (SEQ ID NO: 46), (35) pGlu-Arg-Pro-Arg-Leu-Ser-His
-Lys-Gly-Pro-Cha-Pro-Phe (SEQ ID NO: 47), (36)
Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-C
ha (SEQ ID NO: 48), (37) Arg-Arg-Gln-Arg-Pro-A
rg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe (Cl) (SEQ ID NO: 49), (38) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser
-His-Nle-Gly-Pro-Met-Pro-Phe (Cl) (SEQ ID NO: 5)
0), (39) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Nle
-Gly-Pro-Met-Pro-Tyr (I) (SEQ ID NO: 51), and (40) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Nle-Gly
-Pro-Met-Pro-Tyr (Me) (SEQ ID NO: 52) and the like.

The modified form (II) of apelin has the formula P1-Ar
g-Pro-Arg-Leu-Phe-P2-P3-G
With ly-Pro-P4-P5, a modified form of apelin (II
I) is of the formula Q1-Arg-Pro-Arg-Leu-S.
er-Ala-Q2-Gly-Q5-Q3-Q4, the modified form (IV) of apelin has the formula R1-Arg-Pro-A.
rg-Leu-Ser-His-Lys-Gly-Pr
Represented by o-R2-Pro-R3, each amino acid residue (side chain) in the formula may be substituted, and examples of the substituent include the above-mentioned substituents and the like. In the present specification, P1, Q1 and R1 represent “a hydrogen atom or an amino acid residue or a peptide chain consisting of 1 to 25 amino acids which may be the same or different and whose side chains may be substituted”. Examples of the substituent in the “1 to 25 amino acids whose side chain may be substituted” are the same as the above “Substituents which may be substituted in the side chain of an amino acid residue”. Can be given. Preferred examples when P1, Q1 and R1 represent “an amino acid residue whose side chain may be substituted” include, for example, optionally substituted pyroglutamic acid or optionally substituted side chain glutamine. And the like, and more preferably pyroglutamic acid or glutamine. The “amino acid residue whose side chain may be substituted”,
Examples of the substituent of the amino acid residue in the “optionally substituted pyroglutamic acid” and “the side chain may be substituted glutamine” include, for example, the above “the side chain of the amino acid residue may be substituted. The same as "good substituents" can be mentioned. Moreover, a benzyloxycarbonyl group etc. are mentioned as a preferable substituent of "pyroglutamic acid which may be substituted". Specific examples when P1, Q1 and R1 represent “a peptide chain consisting of 2 to 25 amino acids which may be the same or different and whose side chains may be substituted” include, for example, the formula Y ¹ -Y ² ( In the formula, Y ¹ represents an amino acid residue or a peptide chain consisting of 1 to 17 amino acids in which the side chains may be the same or different and the side chains may be substituted, and Y ² is the same or different and the side chains are substituted. A peptide represented by an amino acid residue or a peptide chain consisting of 1 to 8 amino acids which may be present. Examples of the substituent on the side chain of the amino acid residue represented by Y ¹ and Y ² or the amino acid residue in the peptide chain include the above-mentioned “substituent which may be substituted on the side chain of the amino acid residue”. And the like.

Y above¹For example,
For example, formula (a) A¹-A^Two-A^Three-A^Four-A⁵-A⁶-A
⁷-A⁸-A⁹-A¹⁰-A ¹¹-A¹²-A^Thirteen-A
¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, A¹Through A
¹⁷Have the same or different side chains
An amino acid residue which may be present), (b) Formula A^Two-A^Three−
A^Four-A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰-A¹¹
-A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as described above), (c) formula
A^Three-A^Four-A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰−
A¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as described above), (d) formula
A^Four-A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰-A¹¹
-A¹²-A^{1 Three}-A¹⁴-A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as described above), (e) formula
A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰-A¹¹-A
¹²-A^Thirteen-A ¹⁴-A¹⁵-A¹⁶-A¹⁷(formula
Where each symbol has the same meaning as above), (f) Formula A⁶-A
⁷-A⁸-A⁹-A¹⁰-A¹¹-A¹²-A^Thirteen-A
¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol is as described above.
Have the same meaning), (g) Formula A⁷-A⁸-A⁹-A¹⁰−
A¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A
¹⁷(In the formula, each symbol has the same meaning as described above), (h) formula
A⁸-A⁹-A¹⁰-A¹¹-A¹²-A^Thirteen-A¹⁴
-A¹⁵-A^{1 6}-A¹⁷(In the formula, each symbol is the same as above.
Meaning)), (i) Formula A⁹-A¹⁰-A¹¹-A¹²−
A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A ¹⁷(In the formula, each note
No. has the same meaning as above), (j) Formula A¹⁰-A¹¹−
A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷(formula
Where each symbol has the same meaning as above), (k) Formula A¹¹−
A¹²-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷(formula
Where each symbol has the same meaning as above), (l) Formula A¹²−
A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷(In the formula, each note
No. has the same meaning as above), (m) Formula A^Thirteen-A¹⁴−
A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol has the same meaning as above.
), (N) Formula A¹⁴-A¹⁵-A¹⁶-A
¹⁷(Wherein each symbol has the same meaning as described above), (0) formula
A¹⁵-A¹⁶-A¹⁷(In the formula, each symbol has the same meaning as above.
), (P) Formula A¹⁶-A¹⁷(In the formula, each symbol is
And (q) A¹⁷(A¹⁷Is as above
Amino acid residue or peptide represented by
Examples include chains.

The above A ¹ represents an amino acid residue whose side chain may be substituted, preferably an amino acid residue having an aromatic side chain, more preferably L-having an aromatic side chain. Amino acid residues, more preferably L-tyrosine and the like are shown. The above A ² and A ³ are the same or different and each represents an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably , L-neutral amino acid residue whose side chain may be substituted, more preferably, A ² is L-leucine and the like, and A ³ is L-valine and the like. The above A ⁴ represents an amino acid residue whose side chain may be substituted, preferably a neutral or basic amino acid residue whose side chain may be substituted, more preferably a side chain substituted. Optionally L-neutral or L-basic amino acid residue, more preferably L-lysine or N ^ε -acetyllysine. The above A ⁵ represents an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably L-which may be substituted. Proline, more preferably L-proline and the like are shown. The above A ⁶ and A ⁹ are the same or different and each represent an amino acid residue whose side chain may be substituted, preferably a basic amino acid residue whose side chain may be substituted, and more preferably , L-basic amino acid residue whose side chain may be substituted, more preferably L-arginine. The above A ⁷ and A ¹⁰ are the same or different and each represent an amino acid residue whose side chain may be substituted, but preferably, an amino acid residue having a hydroxy group in the side chain or a side chain is substituted. It also indicates a neutral amino acid residue. As A ⁷ , glycine which may be substituted, particularly glycine, is preferably used. The above A ⁸ represents an amino acid residue whose side chain may be substituted, preferably an amino acid residue having L-proline or a hydroxy group in its side chain, more preferably L-serine, L
-Indicates proline or oxyproline (hydroxyproline) and the like. The above A ¹⁰ is preferably an amino acid residue having a hydroxy group in its side chain or a neutral amino acid residue whose side chain may be substituted, more preferably serine, threonine, asparagine and the like. A ¹¹ above
To A ¹⁴ are the same or different and each represent an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably A ¹¹ Are glycine, A ¹² is L-proline, A ¹³ is glycine, and A ¹⁴ is L-alanine, L-proline and the like. The above A ¹⁵ represents an amino acid residue whose side chain may be substituted, preferably an amino acid residue having an aromatic side chain, more preferably an L-amino acid having an aromatic side chain. Residues, more preferably L-tryptophan and the like are shown. The above A ¹⁶ represents an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably a neutral L- having a carbamoyl group. Amino acid residue, more preferably L-
Indicates glutamine. Examples of the neutral L-amino acid residue having a carbamoyl group include L-glutamine and L-asparagine. A ¹⁷ represents an amino acid residue whose side chain may be substituted, preferably a neutral amino acid residue whose side chain may be substituted, more preferably glycine and the like.

Y above^TwoFor example,
If  Formula B¹-B^Two-B^Three-B^Four-B⁵-B⁶-B⁷-B
⁸(In the formula, B¹Through B ⁸Are the same or different
Represents an amino acid residue whose side chain may be substituted),  Formula B^Two-B^Three-B^Four-B⁵-B⁶-B⁷-B⁸(formula
, Each symbol has the same meaning as above),  Formula B^Three-B^Four-B⁵-B⁶-B⁷-B⁸(Each in the formula
The symbols have the same meaning as above),  Formula B^Four-B⁵-B⁶-B⁷-B⁸(In the formula, each symbol is
(Same meaning as above),  Formula B⁵-B⁶-B⁷-B⁸(In the formula, each symbol is as above.
Have the same meaning),  Formula B⁶-B⁷-B⁸(In the formula, each symbol has the same meaning as above.
),  Formula B⁷-B⁸(In the formula, each symbol has the same meaning as above.
Or)  Formula B⁸(B⁸Represents the same meaning as above)
Examples include mino acid residues and peptide chains. B above¹Is an amino acid residue whose side chain may be substituted
Although shown, preferably, the side chain may be substituted neutral
An amino acid residue, more preferably a side chain substituted
A neutral L-amino acid residue, more preferably a substitution
Optionally glycine, most preferably glycine
Etc. B above^TwoThrough B^FourAre the same or
Differently, the amino acid residue whose side chain may be substituted
Although shown, preferably, a base whose side chain may be substituted
Amino acid residues, more preferably side chains substituted
Optionally a basic L-amino acid residue, more preferably
B^TwoAs L-arginine, B^ThreeAs L-Argi
Nin, B^FourShows L-lysine and the like. B above⁵
Represents an amino acid residue whose side chain may be substituted,
Preferably, an amino acid residue having an aromatic side chain,
More preferably, the L-amino acid residue having an aromatic side chain
Group, more preferably L-phenylalanine
You B above⁶And B⁷Are the same or different
Indicates an amino acid residue whose side chain may be substituted
However, preferably, a basic group in which a side chain may be substituted is used.
Minoic acid residue, more preferably even if the side chain is substituted
A good basic L-amino acid residue, more preferably L-
Indicates arginine, etc. B above⁸Has side chains replaced
Amino acid residue which may be present, but preferably the side chain is
Optionally substituted glutamine, more preferably L-
Indicates glutamine, etc.

Y¹And Y^TwoAs a combination of
When expressed by (a) expression + above expression
(Ie X¹But A ¹-A^Two-A^Three-A^Four-A⁵-A⁶−
A⁷-A⁸-A⁹-A¹⁰-A¹¹-A^{1 Two}-A^Thirteen−
A¹⁴-A¹⁵-A¹⁶-A¹⁷-B¹-B^Two-B^Three−
B^Four-B⁵-B⁶-B⁷-B⁸When represented by:
It is omitted in the explanation of the combination), and is shown in (a) above.
When expressed by the above formula + the above formula, the above
(a) + the above expression
If it is expressed as (a) above + the above expression
If the above is true, the formula represented by (a) above + the formula represented by above
When expressed by, the expression expressed in (a) above + expressed in the above
When expressed by the formula, the formula expressed by (a) above + the above
When expressed by the formula, the formula expressed by (a) above +
When represented by the formula represented above, represented by (b) above
When expressed by the formula + the above formula, in (b) above
When expressed by the expression + the above expression, the above
(b) + the above expression
In this case, the formula expressed by (b) above + the formula expressed by above
If the above is true, the formula expressed in (b) above + the formula expressed above
When expressed by, the formula expressed by (b) above + expressed by the above
When expressed by the formula, the formula expressed by (b) above + the above
When expressed by the formula, the formula expressed by (b) above +
When represented by the formula represented above, represented by the above (c)
In case of the formula + the formula above, in (c) above
When expressed by the expression + the above expression, the above
(c) + (expression above)
In this case, it is represented by the formula (c) above + the formula above
If the above is true, the formula represented by (c) above + the formula represented by above
When expressed by, the formula expressed by (c) above + expressed by the above
When expressed by the formula, the formula expressed by (c) above + the above
When expressed by the formula, the formula expressed by (c) above +
When represented by the formula represented above, it is represented by (d) above.
In the case of the formula + the formula shown above, in (d) above
When expressed by the expression + the above expression, the above
(d) + the above expression
In this case, it is represented by the formula represented by (d) above + the formula represented by above
In case of the above, the formula represented by the above (d) + the formula represented by the above
When expressed by, the formula expressed by (d) above + expressed by the above
When expressed by the formula, the formula expressed by (d) above + the above
When expressed by the formula, the formula expressed by (d) above +
When represented by the formula represented above, it is represented by (e) above.
In case of the formula + the formula shown above, in (e) above
When expressed by the expression + the above expression, the above
(e) + the above expression
In this case, the formula expressed by (e) above + the formula expressed by above
If the above is true, the formula expressed in (e) above + the formula expressed above
When expressed by, the expression expressed by (e) above + expressed by the above
When expressed by the formula, the formula expressed by (e) above + the above
When expressed by the formula, the formula expressed by (e) above +
When represented by the formula represented above, it is represented by (f) above.
In the case of being expressed by the formula + the above formula, in (f) above
When expressed by the expression + the above expression, the above
(f) + the above expression
In this case, it is expressed by the formula shown in (f) above + the formula shown above
In case of the above, the formula represented by the above (f) + the formula represented by the above
When expressed by, the formula expressed by (f) above + expressed by
When expressed by the formula, the formula expressed by (f) above + the above
When expressed by the formula, the formula expressed by (f) above +
When represented by the formula represented above, represented by (g) above
Formula + the above formula, in (g) above
When expressed by the expression + the above expression, the above
(g) + the above expression
If it is expressed by the formula (g) above + the formula above
If the above is true, the formula represented by the above (g) + the formula represented by the above
When expressed by, the formula expressed by (g) above + expressed by
When expressed by the formula, the formula expressed by (g) above + the above
When expressed by the formula, the formula expressed by (g) above +
When represented by the formula represented above, it is represented by (h) above.
In the case of being expressed by the formula + the above formula, in (h) above
When expressed by the expression + the above expression, the above
(h) + the above expression
In this case, it is expressed by the formula represented by (h) + the formula represented by the above.
In case of the above, the formula represented by the above (h) + the formula represented by the above
When expressed by, the formula expressed by (h) above + expressed by the above
When expressed by the formula, the formula expressed by (h) above + the above
When expressed by the formula, the formula expressed by (h) above +
When represented by the formula represented above, represented by (i) above
In the case of the above formula + the above formula, in (i) above
When expressed by the expression + the above expression, the above
(i) + the above expression
In this case, it is represented by the formula (i) above + the formula above
If the above is true, the formula represented by (i) above + the formula represented by above
When expressed by, the expression expressed by (i) above + expressed by the above
When expressed by the formula, the formula expressed by (i) above + the above
When expressed by the formula, the formula expressed by (i) above +
When represented by the above formula, it is represented by (j) above.
Formula + the above formula, in (j) above
When expressed by the expression + the above expression, the above
(j) + (expression above)
If it is expressed as (j) above + the above expression
If the above is true, the formula expressed in (j) above + the formula expressed above
When expressed by, the expression expressed by (j) above + expressed by the above
When expressed by the formula, the formula expressed by (j) above + the above
When expressed by the formula, the formula expressed by (j) above +
When represented by the formula represented above, it is represented by (k) above.
In the case of being expressed by the formula + the above formula, in (k) above
When expressed by the expression + the above expression, the above
(k) + the above expression
If it is expressed by the above formula (k) + the above formula,
In case of the above, the formula represented by the above (k) + the formula represented by the above
When expressed by, the expression expressed by (k) above + expressed by the above
When expressed by the formula, the formula expressed by (k) above + the above
When expressed by the formula, the formula expressed by (k) above +
When represented by the formula represented above, it is represented by (l) above.
Formula + the above formula, in (l) above
When expressed by the expression + the above expression, the above
(l) + the above expression
The formula expressed by (l) above + the formula expressed by above
If the above is true, the formula represented by (l) above + the formula represented above
When expressed by, the formula expressed by (l) above + the above expressed
When expressed by the formula, the formula expressed by (l) above + the above
When expressed by the formula, the formula expressed by (l) above +
When represented by the formula represented above, it is represented by (m) above.
In the case of being expressed by the formula + the above formula, in (m) above
When expressed by the expression + the above expression, the above
(m) + the above expression
If it is expressed by the formula (m) above + the formula above
In case of the above, the formula represented by the above (m) + the formula represented by the above
When expressed by, the formula expressed by (m) above + expressed by the above
When expressed by the formula, the formula expressed by (m) above + the above
When expressed by the formula, the formula expressed by (m) above +
When represented by the formula represented above, represented by (n) above
When expressed by the formula + the formula above, in (n) above
When expressed by the expression + the above expression, the above
(n) + the above expression
In this case, it is represented by the formula represented by (n) above + the formula represented by above
If the above is true, the formula represented by (n) above + the formula represented above
Is expressed by the above formula (n) + the above
When expressed by the formula, the formula expressed by (n) above + the above
When expressed by the formula, the formula expressed by (n) above +
When represented by the formula represented above, it is represented by (o) above.
In case of the formula + the formula shown above, the above (o)
When expressed by the expression + the above expression, the above
(o) + the above expression
If it is expressed as (o) above + the above expression
If the above is true, the formula represented by (o) above + the formula represented by above
When expressed by, the expression expressed by (o) above + expressed by the above
When expressed by the formula, the formula expressed by (o) above + the above
When expressed by the formula, the formula expressed by (o) above +
When represented by the formula represented above, it is represented by (p) above.
In the case of the formula + the formula shown above, in (p) above
When expressed by the expression + the above expression, the above
(p) + the above expression
In this case, it is expressed by the formula (p) above + the formula above
In case of the above, the formula represented by the above (p) + the formula represented by the above
When expressed by, the expression expressed by (p) above + expressed by the above
When expressed by the formula, the formula expressed by (p) above + the above
When expressed by the formula, the formula expressed by (p) above +
When expressed by the above formula, it is expressed by (q) above.
In the case of the formula + the formula shown above, the above (q)
When expressed by the expression + the above expression, the above
(q) + the above expression
If it is expressed as (q) above + (expression above)
In case of the above, the formula represented by the above (q) + the formula represented by the above
When expressed by, the expression expressed by (q) above + expressed by the above
When expressed by the formula, the formula expressed by (q) above + the above
When represented by the formula represented by, and represented by (q) above
There are cases where it is expressed by the formula + the formula shown above.
But,

Above all, the formula represented by the above (a) + the above
When expressed by the formula, the formula expressed by (b) above +
When represented by the formula represented above, represented by the above (c)
In the case of the formula + the formula shown above, in (d) above
When expressed by the expression + the above expression, the above
(e) + the above expression
In this case, it is expressed by the formula shown in (f) above + the formula shown above
If the above is true, the formula represented by the above (g) + the formula represented by the above
When expressed by, the formula expressed by (h) above + expressed by the above
When expressed by the formula, the formula expressed by (i) above + the above
When expressed by the formula, the formula expressed by (j) above +
When represented by the formula represented above, it is represented by (k) above.
Formula + the above formula, in (l) above
When expressed by the expression + the above expression, the above
(m) + the above expression
In this case, it is represented by the formula represented by (n) above + the formula represented by above
If the above is true, the formula represented by (o) above + the formula represented by above
When expressed by, the expression expressed by (p) above + expressed by the above
When expressed by the formula, and the formula expressed by (q) above +
The case represented by the formula shown above is preferable. in
Is also expressed by the formula shown in (a) above + the formula shown above
And the formula shown in (b) above + the above
The case represented by the formula
It Expressed by the formula shown in (a) above + the formula shown above
And the formula shown in (b) above + the above
A more preferable specific example of the case represented by the formula
You Expressed by the formula shown in (b) above + the formula shown above
And P1, Q1 and / or R1 are of the formula A
^Two-A^Three-A^Four-A⁵-A⁶-A⁷-A⁸-A⁹-A
¹⁰-A¹¹-A¹²-A^Thirteen-A¹⁴-A¹⁵-A
¹⁶-A¹⁷-B¹-B ^Two-B^Three-B^Four-B⁵-B⁶−
B⁷-B⁸(In the formula, A²Through A¹⁷And B¹No
B⁸Has the same meaning as above).
However, a preferred specific example in this case is A^Twoand
A^ThreeAre the same or different and the side chains are
Optional L-neutral amino acid residue, preferably A^Twoage
Such as L-leucine, A^ThreeAs for L-valine etc.
Yes, A^FourIs an L-neutral optionally substituted side chain,
Is L-basic amino acid residue, preferably L-glutamine
L-lysine or N^ε-Such as acetyllysine
A⁵Is L-proline in which the side chain may be substituted,
Preferred is L-proline or the like, and A⁶And A⁹
Are the same or different and the side chains are
Good L-basic amino acid residue, preferably L-argi
Such as nin, A⁷Glycy optionally substituted
A, preferably glycine and the like, A⁸Is L-Proli
Amino acid residues having side chains or hydroxyl groups,
Preferably, L-serine, L-proline or oxypro
Such as phosphorus (hydroxyproline), A¹⁰But hi
Amino acid residue or side chain having droxy group
An optionally substituted neutral amino acid residue, preferably L-
Serine, L-threonine, L-asparagine, etc.
A¹¹Is glycine, A¹²Is L-proline, A^Thirteen
Is glycine, A¹⁴Is L-alanine or L-proline
What is A¹⁵L-amino having an aromatic side chain
Acid residue, preferably L-tryptophan and the like,
¹⁶Is a neutral L-amino acid residue having a carbamoyl group,
More preferably L-glutamine, A¹⁷Is a neutral amino acid
A residue, preferably glycine and the like, B¹Has side chains
An optionally substituted neutral L-amino acid residue, preferably
Is an optionally substituted glycine, more preferably,
Such as glycine, B^TwoThrough B^FourAre the same
Alternatively, a basic L-, in which the side chain may be substituted,
Amino acid residue, preferably B^TwoAs L-Arginii
B,^ThreeAs L-arginine, B^FourAs L-
Such as lysine, B⁵L-has an aromatic side chain
Amino acid residue, preferably L-phenylalanine etc.
And B⁶And B⁷Are the same or different
And a basic L-amino acid residue whose side chain may be substituted.
A group, preferably L-arginine and the like, B⁸Is on the side
Glutamine whose chain may be substituted, preferably L
-Examples include glutamine.

Formula represented by (a) above + represented by above
When expressed by a formula, X¹Is the expression A ¹-A^Two-A^Three−
A^Four-A⁵-A⁶-A⁷-A⁸-A⁹-A¹⁰-A¹¹
-A^{1 Two}-A^Thirteen-A¹⁴-A¹⁵-A¹⁶-A¹⁷−
B¹-B^Two-B^Three-B^Four-B⁵-B⁶-B⁷-B⁸(formula
Medium, A¹Through A¹⁷And B¹Through B⁸Is the same as above
In this case,
As a preferred specific example of¹Has aromatic side chains
L-amino acid residue, more preferably L-tyrosine
And A^TwoAnd A^ThreeAre the same or different
And an L-neutral amino acid residue whose side chain may be substituted
A group, preferably A^TwoAs L-leucine, A^ThreeWhen
Is L-valine, and A^FourHas side chains replaced
L-neutral or L-basic amino acid residue which may be
More preferably, L-glutamine, L-α aminoadipic acid, L-
Lysine or N^ε-Acetyllysine, etc., A⁵But
Optionally substituted L-proline, preferably L-proline
Such as Lorin, A⁶And A⁹Are the same
Are different from each other, and L-basic acid optionally substituted in the side chain.
A mino acid residue, preferably L-arginine and the like, A
⁷Optionally substituted glycine, preferably glycine
A, etc., and⁸Is L-proline or hydroxy group
An amino acid residue having a side chain, preferably L-seri
, L-proline or oxyproline (hydroxyproline
Lorin) etc., and A¹⁰Has a hydroxy group in the side chain
The amino acid residue or side chain to
Amino acid residue, preferably L-serine, L-threoni
A, L-asparagine, etc.¹¹Is glycine,
A¹²Is L-proline, A^ThirteenIs glycine, A¹⁴Is L-
Such as alanine or L-proline, A¹⁵Is fragrance
L-amino acid residue having an aromatic side chain, preferably L-
Tryptophan, etc., A¹⁶Is a carbamoyl group
Having a neutral L-amino acid residue, preferably L-glutamine
And A¹⁷Is a neutral amino acid residue, preferably
Shin, etc., B¹In which the side chain may be substituted
Sex L-amino acid residue, preferably optionally substituted
Glycine, more preferably glycine and the like, B
^TwoThrough B^FourAre the same or different and the side chains are
A basic L-amino acid residue which may be replaced, preferably
Is B^TwoAs L-arginine, B^ThreeAs L-a
Luginin, B^FourIs L-lysine or the like, and B⁵
Is an L-amino acid residue having an aromatic side chain, preferably
Is L-phenylalanine or the like, and B⁶And B⁷
Are the same or different and the side chains are
A basic L-amino acid residue, preferably L-alcohol
Guinin, etc., B⁸May have a side chain substituted
Glutamine, preferably L-glutamine L-pyrogluta
Examples include cases where it is a mycotic acid.

Preferred components for P1, Q1 and R1
As body examples, (1) hydrogen atom, (2) Arg-Gln, (3) Arg, (4)
Gln, (5) pGlu, or (6) Arg-Arg-Gln.
It Further, P1 is a hydrogen atom, pGlu or
Arg-Arg-Gln is more preferable, and Q1 is hydrogen atom.
Child, pGlu or Arg-Arg-Gln is more preferred, and R1
Is a hydrogen atom, pGlu or Arg-Arg-Gln (further
Is more preferably a hydrogen atom or Arg-Arg-Gln). True spirit
In the detailed text, P2 is a neutral group whose side chain may be substituted.
Basicity in which amino acid residues or side chains may be substituted
Indicates an amino acid residue, preferably substituted (side chain)
Optionally L-alanine, substituted (side chain)
Moyoi L-histidine and the like can be mentioned. In this specification
And side chains of P3 and Q2 may be substituted.
Aromatic amino acid residue, aromatic group which side chain may be substituted
Minoic acid residue or basic that side chain may be substituted
Indicates an amino acid residue. Placement of a basic amino acid residue on the side chain
Examples of the substituent include C_1-4Acyl group, tosyl group, C
_1-6Examples thereof include an alkyl group. C_1-4Acyl group
As, for example, formyl, acetyl, propionyl,
Butyryl and C_2-4Such as alkanoyl
Be done. C_1-6Examples of the alkyl group include methyl and ether.
Chill, propyl, isopropyl, butyl, isobutyl,
sec-butyl, tert-butyl, pentyl, hexyl etc.
Can be given. P3 and Q2 are preferably side chains
L-lysine optionally substituted, side chain substituted
L-norleucine which may be present, or a side chain is substituted
L-arginine, etc., which can be used, are preferred and more preferred.
The side chain is C_1-4Acyl group (eg formyl, acetyl
And propionyl, butyryl and C and_2-4Arca
Noil), C_1-6Alkyl group (eg, methyl, ethyl
Le, propyl, isopropyl, butyl, isobutyl, se
(C-butyl, tert-butyl, pentyl, hexyl etc.)
Or L-lysine optionally substituted with a tosyl group, L
-Norleucine or L-arginine and the like,
More preferably, L-lysine, L-norleucine, L
-Arginine, N^ε-Acetyllysine, N^ε-Methyl
Jin, N ^ε-Tosyl lysine, N^g-Tosyl arginine
And so on. Furthermore, as P3 and Q2,
Optionally substituted L-arginine or substituted
Optional L-lysine is preferred. In this specification,
P4 and Q3 may have a bond or side chain substituted
Aromatic amines with good neutral or optionally substituted side chains
No acid residue is shown. P4 and Q3 are preferably
L-norleucine in which a bond or side chain may be substituted
L-methionine in which side chain may be substituted, side chain
L-methionine sulfoxide which may be substituted
Or L-alanine optionally substituted in the side chain
And more preferably a bond, L-norleucine,
L-methionine, L-methionine sulfoxide or
L-cyclohexylalanine and the like can be mentioned. Furthermore
In addition, as P4, L-norleucine, L-methionine
Or L-cyclohexylalanine is particularly preferable.
Yes. Further, as Q3, a bond, L-methionine or
Is particularly preferably L-cyclohexylalanine. Book
In the specification, P5 and Q4 have side chains substituted
Optional amino acid residue or its C-terminal carboxyl
Group reduced to hydroxymethyl or formyl group
Amino acid derivative, hydroxyl group or side chain is substituted
Optional amino acid residue and side chain may be substituted
Dipeptide chain formed by binding mino acid residue or C-terminal thereof
If the end carboxyl group is a hydroxylmethyl group or
Shows a peptide derivative reduced to a phenyl group.

P5 and Q4 are preferably neutral amino acid residues whose side chains may be substituted or Cs thereof.
Amino acid derivative whose terminal carboxyl group is reduced to hydroxymethyl group or formyl group, neutral amino acid residue which may be substituted on hydroxyl group or side chain and aromatic side chain which may be substituted on side chain Examples thereof include a dipeptide chain having an amino acid residue bound thereto or a peptide derivative in which the C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group. More preferably as P5 and Q4, L-proline optionally substituted in the side chain or an amino acid derivative in which the C-terminal carboxyl group thereof is reduced to a hydroxymethyl group or a formyl group, or the side chain may be substituted 4 -Amino acid derivative in which chlorophenylalanine or its C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, 2-naphthylalanine whose side chain may be substituted or its C-terminal carboxyl group is converted to a hydroxymethyl group or a formyl group A reduced amino acid derivative, an amino acid derivative in which a C-terminal carboxyl group of cyclohexylalanine whose side chain may be substituted is reduced to a hydroxymethyl group or a formyl group,
Hydroxyl group or optionally substituted L-proline and
(a) L-phenylalanine whose side chain may be substituted, (b) L-tyrosine whose side chain may be substituted, (c)
L-2-thienylalanine whose side chain may be substituted, (d) L-phenylglycine whose side chain may be substituted, or (e) whose side chain may be substituted L-
Examples thereof include a dipeptide chain formed by binding 2-pyridylalanine, or a peptide derivative in which the C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group. More preferably as P5 and Q4,
(1) Amino acid derivative in which L-proline or its C-terminal carboxyl group is reduced to a hydroxymethyl group or formyl group, (2) 4-chlorophenylalanine or its C-terminal carboxyl group is reduced to a hydroxymethyl group or formyl group Amino acid derivative, (3) Amino acid derivative in which 2-naphthylalanine or its C-terminal carboxyl group is reduced to hydroxymethyl group or formyl group, (4) Cyclohexylalanine or its C-terminal carboxyl group is reduced to hydroxymethyl group or formyl group Amino acid derivative, (5) hydroxyl group, (6) dipeptide chain formed by binding L-proline and L-phenylalanine or a peptide derivative in which its C-terminal carboxyl group is reduced to a hydroxylmethyl group or a formyl group, (7) L
-A peptide derivative in which proline and L-tyrosine are bound or a peptide derivative in which the C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group, (8)
(9) L-proline bound to L-2-thienylalanine or a peptide derivative having a C-terminal carboxyl group reduced to a hydroxylmethyl group or a formyl group, (9) L-proline bound to L-phenylglycine Or a peptide derivative in which the C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group, (10) L-proline and 4-chlorophenylalanine bound to the dipeptide chain or its C-terminal carboxyl group Peptide derivative reduced to a hydroxylmethyl group or formyl group, (11) Peptide derivative in which L-proline and 2-naphthylalanine are bound or a C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group , (12)
A dipeptide chain consisting of L-proline and 3-iodotyrosine bound or a peptide derivative whose C-terminal carboxyl group is reduced to a hydroxylmethyl group or formyl group, (13) L-proline and O-methyltyrosine bound Peptide derivative or a peptide derivative in which the C-terminal carboxyl group thereof is reduced to a hydroxylmethyl group or a formyl group, or (14) a dipeptide chain formed by binding L-proline and L-2-pyridylalanine or its C-terminal carboxyl group Examples thereof include peptide derivatives in which is reduced to a hydroxylmethyl group or a formyl group.

"-P4-P5" and "-Q3-Q4"
As particularly preferable, (1) -Nle-Pro-Phe, (2) -Nle
-Pro-Tyr, (3) -Nle-Pro, (4) -Nle, (5) -Met-Pro-Ph
e, (6) -Nle-Pro-Thi, (7) -Nle-Pro-Phg, (8) -Nle-Pr
o-Pya (2), (9) -Met (O), (10) -Met-Phe (Cl), (11) -Me
t-Pro-Phe (Cl), (12) -Met-Pro-Nal (2), (13) -Met-Nal
(2), (14) -Met-Cha, (15) -Cha-Pro-Phe, (16) -Cha,
(17) -Met-Pro-Tyr (I), (18) -Cha-Pro-Phe (Cl), (19)
-Cha-Phe (Cl), (20) -Nle-Pro-Tyr (I), (21) -Nle-Phe
(Cl), (22) -Cha-Pro-Tyr (I), (23) -Cha-Tyr (I), (24)
-Cha-Phe, (25) -Met-Phe, (26) -Met-Pro-Tyr (Me),
(27) -OH, (28) -Met, (29) -Met-Pro-Phe, and (30)
-Ala-Pro-Phe (Cl) etc. are mentioned. Also, "-P4
-P5 "means (1) -Cha-Pro-Phe (Cl), (2) -Cha-P
ro-Phe, (3) -Met-Pro-Phe (Cl), (4) -Met-Pro-Phe (5)
-Cha-Phe, and (6) -Met-Phe are preferable, and (1) -Cha-P
ro-Phe (Cl), (2) -Cha-Pro-Phe, (3) -Met-Pro-Phe (C
l) and (4) -Met-Pro-Phe are particularly preferred. further,
"-Q3-Q4" includes (1) -Met-Pro-Phe (Cl),
(2) -Cha-Phe (Cl), (3) -Cha-Pro-Phe (Cl), (4) -Cha,
(5) -Cha-Pro-Phe, (6) -OH, (7) -Met, (8) -Met-Pro-
Phe and (9) -Ala-Pro-Phe (Cl) are especially preferred.

In the present specification, Q5 represents a neutral amino acid residue whose side chain may be substituted, preferably L-proline which may be substituted, L-glycine which may be substituted or This shows L-alanine which may be substituted. Particularly preferably as Q5, L-proline,
Indicates L-glycine or L-alanine. In the present specification, R2 represents optionally substituted L-cyclohexylalanine, preferably L-cyclohexylalanine. In the present specification, R3 is an optionally substituted L-phenylalanine, an optionally substituted L-
2-naphthylalanine, optionally substituted L-cyclohexylalanine or optionally substituted tyrosine is shown, and preferably L-4-chlorophenylalanine, L-2-naphthylalanine, L-cyclohexylalanine, L-phenylalanine. Alternatively, it represents L-tyrosine.

Specific examples of modified apelin (II)
Is, for example, (1) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-A
la-Arg-Gly-Pro-Met-Pro-Phe (Cl), (2) Arg-Arg-Gln-A
rg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Cha-Pro-Phe (C
l), (3) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Met-P
ro-Phe (Cl), (4) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-P
ro-Cha-Phe (Cl), (5) Arg-Pro-Arg-Leu-Ser-Ala-Arg-G
ly-Pro-Cha-Pro-Phe (Cl), (6) Arg-Arg-Gln-Arg-Pro-A
rg-Leu-Ser-Ala-Arg-Gly-Pro-Cha, (7) Arg-Pro-Arg-L
eu-Ser-His-Lys-Gly-Pro-Cha-Pro-Phe (Cl), (8) Arg-P
ro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe (Cl),
(9) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Cha-
Pro-Phe, (10) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gl
y-Pro-Met-Pro-Phe (Cl), (11) Arg-Pro-Arg-Leu-Ser-A
la-Arg-Gly-Pro-Cha-Pro-Phe, (12) Arg-Pro-Arg-Leu-
Phe-Ala-Arg-Gly-Pro-Cha-Pro-Phe (Cl), (13) Arg-Pro
-Arg-Leu-Phe-His-Lys-Gly-Pro-Cha-Pro-Phe (Cl), (14)
  Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-
Nle-Pro-Tyr, (15) Arg-Pro-Arg-Leu-Phe-His-Lys-Gly
-Pro-Cha-Pro-Phe, (16) Arg-Pro-Arg-Leu-Phe-His-Ly
s-Gly-Pro-Met-Pro-Phe (Cl), (17) Arg-Arg-Gln-Arg-P
ro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Met-Pro-Phe, (18)
Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Me
t, (19) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-Ala-Arg-G
ly-Pro, (20) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-
Lys-Gly-Pro-Met (O), (21) Arg-Arg-Lys (Arg-Arg) -A
rg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe, (2
2) Arg-Arg-Arg-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pr
o-Met-Pro-Phe, (23) Arg-Arg-Lys-Arg-Pro-Arg-Leu-S
er-His-Lys-Gly-Pro-Met-Pro-Phe, (24) Arg-Pro-Arg-
Leu-Ser-Ala-Arg-Gly-Pro-Met-Pro-Phe, (25) Arg-Arg
-Ala-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-P
he, (26) pGlu-Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro
-Met-Pro-Phe, (27) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gl
y-Pro-Met, (28) Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-P
ro-Met-Phe (Cl), (29) pGlu-Arg-Pro-Arg-Leu-Ser-Ala
-Lys-Gly-Pro-Met-Pro-Phe, (30) pGlu-Arg-Pro-Arg-L
eu-Ser-Arg-Lys-Gly-Pro-Met-Pro-Phe, (31) Arg-Pro-
Arg-Leu-Phe-Ala-Arg-Gly-Pro-Met-Pro-Phe, (32) Arg
-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Nal (2),
(33) Arg-Arg-Phe-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-
Pro-Met-Pro-Phe, (34) pGlu-Arg-Pro-Arg-Leu-Ser-Hi
s-Arg-Gly-Pro-Met-Pro-Phe, (35) pGlu-Arg-Pro-Arg-
Leu-Ser-His-Lys-Gly-Pro-Met-Phe (Cl), (36) Arg-Pro
-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Cha, (37) Arg-Pr
o-Arg-Leu-Phe-His-Lys-Gly-Pro-Cha-Phe (Cl), (38) p
Glu-Arg-Pro-Arg-Leu-Ser-Leu-Lys-Gly-Pro-Met-Pro-Ph
e, (39) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-
Met-Pro-Nal (2), (40) Arg-Pro-Arg-Leu-Ser-His-Lys-
Gly-Pro-Met-Nal (2), (41) pGlu-Arg-Pro-Arg-Leu-Ser
-Arg-Arg-Gly-Pro-Met-Pro-Phe, (42) pGlu-Arg-Pro-A
rg-Leu-Phe-Arg-Arg-Gly-Pro-Met-Pro-Phe, (43) pGlu
-Arg-Pro-Arg-Leu-Ser-Phe-Lys-Gly-Pro-Met-Pro-Phe,
(44) pGlu-Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Met
-Pro-Phe, (45) pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-G
ly-Pro-Met-Cha, (46) pGlu-Arg-Pro-Arg-Leu-Ser-His
-Lys-Gly-Pro-Met-Nal (2), (47) Arg-Pro-Arg-Leu-Phe
-Ala-Arg-Gly-Pro-Met-Phe, (48) pGlu-Arg-Pro-Arg-L
eu-Ser-His-Phe-Gly-Pro-Met-Pro-Phe, (49) Arg-Pro-
Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Cha, (50) pGl
u-Arg-Pro-Arg-Leu-Ser-His-Leu-Gly-Pro-Met-Pro-Ph
e, (51) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-Ala-Arg-G
ly-NMe₂, (52) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-Ala
-Arg-Gly-Mor, (53) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gl
y-Pro-Ala-Pro-Phe (Cl), (54) Arg-Pro-Arg-Leu-Ser-A
la-Arg-Gly-Gly-Met-Pro-Phe (Cl), (55) Arg-Pro-Arg-
Leu-Ser-Ala-Arg-Gly-N-MeAla-Met-Pro-Phe (Cl), (56)
  Arg-Pro-Arg-Leu-Ser-His-Ala-Gly-Pro-Cha-Pro-Phe
(Cl), (57) Arg-Pro-Arg-Ala-Ser-His-Lys-Gly-Pro-Ch
a-Pro-Phe (Cl), (58) Arg-Pro-Ala-Leu-Ser-His-Lys-G
ly-Pro-Cha-Pro-Phe (Cl), (59) Arg-Arg-Gln-Arg-Pro-
Arg-Leu-Ser-Ala-Arg-Gly-Pro-Met-Pro-Phe (Cl) -NH ₂,
(60) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Cha-Py
n, (61) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Cha-P
ro-Pyn, (62) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-Ala-
Arg-Gly-Gly-Cha, (63) Arg-Pro-Lys (Me)₂-Leu-Ser-Al
a-Arg-Gly-Pro-Met-Pro-Phe, (64) Arg-Pro-Arg-Leu-S
er-Ala-Lys (Me)₂-Gly-Pro-Met-Pro-Phe, (65) Arg-Pro
-Arg-Leu-Ser-Dap-Arg-Gly-Pro-Cha-Pro-Phe (Cl), (66)
  Arg-Pro-Arg-Leu-Ser-Dap (Ac) -Arg-Gly-Pro-Cha-Pro-
Phe (Cl), (67) Arg-Pro-Arg-Leu-Ser-Dap (C₆) -Arg-Gly
-Pro-Cha-Pro-Phe (Cl), and (68) Arg-Pro-Arg-Leu-
Ser-Dap (Adi) -Arg-Gly-Pro-Cha-Pro-Phe (Cl) etc.
To be

Specific examples of the modified form (III) of apelin include Arg-Pro-Arg-Leu-Phe-Ala-Arg-Gly-Pro-Cha-Pro-Phe.
(Cl), Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Cha-Pro-Phe
(Cl), Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Cha-Pro-Ph
e, Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Met-Pro-Phe
(Cl), Arg-Pro-Arg-Leu-Phe-Ala-Arg-Gly-Pro-Met-Pro-Ph
e, Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Cha-Phe (C
l), pGlu-Arg-Pro-Arg-Leu-Phe-Arg-Arg-Gly-Pro-Met-Pro
-Phe, pGlu-Arg-Pro-Arg-Leu-Phe-His-Lys-Gly-Pro-Met-Pro
-Phe, the peptide represented by Arg-Pro-Arg-Leu-Phe-Ala-Arg-Gly-Pro-Met-Phe, its ester, its salt, etc. are mentioned. Specific examples of modified apelin (IV) include (i) Arg-Ar
g-Gln-Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Met-Pro-
Phe (Cl), (ii) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-
Met-Pro-Phe (Cl), (iii) Arg-Pro-Arg-Leu-Ser-Ala-Arg
-Gly-Pro-Cha-Phe (Cl), (iv) Arg-Pro-Arg-Leu-Ser-Ala
-Arg-Gly-Pro-Cha-Pro-Phe (Cl), (v) Arg-Arg-Gln-Arg-
Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Cha, (vi) Arg-Pro-
Arg-Leu-Ser-Ala-Arg-Gly-Pro-Cha-Pro-Phe, (vii) Arg
-Arg-Gln-Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Met-P
ro-Phe, (viii) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-Ala
-Arg-Gly-Pro-Met, (ix) Arg-Arg-Gln-Arg-Pro-Arg-Leu
-Ser-Ala-Arg-Gly-Pro, (x) Arg-Pro-Arg-Leu-Ser-Ala-
Arg-Gly-Pro-Met-Pro-Phe, (xi) pGlu-Arg-Pro-Arg-Leu
-Ser-Ala-Arg-Gly-Pro-Met-Pro-Phe, (xii) Arg-Pro-Ar
g-Leu-Ser-Ala-Arg-Gly-Pro-Met, (xiii) pGlu-Arg-Pro
-Arg-Leu-Ser-Ala-Lys-Gly-Pro-Met-Pro-Phe, (xiv) Ar
g-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Pro-Ala-Pro-Phe (C
l), (xv) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Gly-Met-P
ro-Phe (Cl), (xvi) Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-
NMe-Ala-Met-Pro-Phe (Cl), (xvii) Arg-Pro-Arg-Leu-Se
r-Ala-Arg-Gly-Pro-Cha-Pyn, (xviii) Arg-Pro-Arg-Leu
-Ser-Ala-Arg-Gly-Pro-Cha-Pro-Pyn, (xix) Arg-Arg-Gl
n-Arg-Pro-Arg-Leu-Ser-Ala-Arg-Gly-Gly-Cha, or
(xx) Arg-Pro-Arg-Leu-Ser-Ala-Lys (Me) ₂ -Gly-Pro-Met-
Examples include the peptide represented by Pro-Phe, its ester, its salt, and the like.

Specific examples of modified apelin (IV) include Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-.
Cha-Pro-Phe (Cl), Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Cha-Pro-Phe
(Cl), Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe
(Cl), pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Cha-Pro
-Phe, pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro
-Phe (Cl), Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro
-Nle-Pro-Tyr, Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Nal
(2), pGlu-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro
-Nal (2), Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Cha
The peptide represented by or its ester, its salt, etc. are mentioned. Furthermore, (i) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-His-Ly
s-Gly-Pro-Met (O), (ii) Arg-Arg-Lys (Arg-Arg) -Arg-Pr
o-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe, (iii) A
rg-Arg-Arg-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met
-Pro-Phe, (iv) Arg-Arg-Lys-Arg-Pro-Arg-Leu-Ser-His
-Lys-Gly-Pro-Met-Pro-Phe, (v) Arg-Arg-Ala-Arg-Pro-
Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Pro-Phe, (vi) Arg-
Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Phe (Cl), (vii)
pGlu-Arg-Pro-Arg-Leu-Ser-Arg-Lys-Gly-Pro-Met-Pro-
Phe, (viii) Arg-Arg-Phe-Arg-Pro-Arg-Leu-Ser-His-Ly
s-Gly-Pro-Met-Pro-Phe, (ix) pGlu-Arg-Pro-Arg-Leu-S
er-His-Arg-Gly-Pro-Met-Pro-Phe, (x) pGlu-Arg-Pro-A
rg-Leu-Ser-His-Lys-Gly-Pro-Met-Phe (Cl), (xi) Arg-P
ro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Cha, (xii) pGlu
-Arg-Pro-Arg-Leu-Ser-Leu-Lys-Gly-Pro-Met-Pro-Phe,
(xiii) Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Nal
(2), (xiv) pGlu-Arg-Pro-Arg-Leu-Ser-Arg-Arg-Gly-Pr
o-Met-Pro-Phe, (xv) pGlu-Arg-Pro-Arg-Leu-Ser-Phe-L
ys-Gly-Pro-Met-Pro-Phe, (xvi) pGlu-Arg-Pro-Arg-Leu
-Ser-His-Lys-Gly-Pro-Met-Cha, (xvii) pGlu-Arg-Pro-
Arg-Leu-Ser-His-Lys-Gly-Pro-Met-Nal (2), (xviii) pG
lu-Arg-Pro-Arg-Leu-Ser-His-Phe-Gly-Pro-Met-Pro-Ph
e, (xix) pGlu-Arg-Pro-Arg-Leu-Ser-His-Leu-Gly-Pro-
Met-Pro-Phe, (xx) Arg-Arg-Gln-Arg-Pro-Arg-Leu-Ser-
Ala-Arg-Gly-NMe ₂ , (xxi) Arg-Arg-Gln-Arg-Pro-Arg-Le
u-Ser-Ala-Arg-Gly-Mor, (xxii) Arg-Pro-Arg-Leu-Ser-
His-Ala-Gly-Pro-Cha-Pro-Phe (Cl), (xxiii) Arg-Pro-A
rg-Ala-Ser-His-Lys-Gly-Pro-Cha-Pro-Phe (Cl), (xxiv)
Arg-Pro-Lys (Me) ₂ -Leu-Ser-Ala-Arg-Gly-Pro-Met-Pro-
Phe, (xxv) Arg-Pro-Arg-Leu-Ser-Dap-Arg-Gly-Pro-Cha
-Pro-Phe (Cl), (xxvi) Arg-Pro-Arg-Leu-Ser-Dap (Ac) c-
Arg-Gly-Pro-Cha-Pro-Phe (Cl), (xxvii) Arg-Pro-Arg-L
eu-Ser-Dap (C6) -Arg-Gly-Pro-Cha-Pro-Phe (Cl), (xxvii
i) Arg-Pro-Arg-Leu-Ser-Dap (Adi) -Arg-Gly-Pro-Cha-Pr
o-Phe (Cl), or (xxix) Arg-Pro-Ala-Leu-Ser-His-Ly
It can also be used for a peptide represented by s-Gly-Pro-Cha-Pro-Phe (Cl), an ester thereof, or a salt thereof. The modified forms of apelin including the modified forms of apelin (I) to (IV) may be an amide form or an ester form in the same manner as the above-mentioned apelin.

As salts of modified apelin (I) to (IV), salts with physiologically acceptable bases (eg alkali metals) and acids (organic acids, inorganic acids) are used.
Particularly, physiologically acceptable acid addition salts are preferable. Examples of such salts include salts with inorganic acids (for example, hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid), or organic acids (for example, acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid, Salts such as tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid) are used. Modified form of apelin (I)
Is a known compound described in WO00 / 18793, and can be produced by the method described in WO00 / 18793. Modified Apelin (II) ~
(IV) is a known compound described in WO01 / 70769, and can be produced by the method described in WO01 / 70769. Hereinafter, the modified form of apelin, the partial peptide thereof, the amide or ester thereof, or the salt thereof may be abbreviated as the modified form of apelin.

As the apelin receptor, its partial peptide or its salt, known ones described in WO00 / 18793 can be used. That is, as the apelin receptor, humans and warm-blooded animals (for example, mammal warm-blooded animals (eg, rabbit, sheep, goat, rat, mouse, guinea pig, cow, horse, pig)), birds (eg, chicken, pigeon, Duck, goose, quail, etc.) (eg, pituitary, pancreas, brain, kidney, liver, gonad, thyroid, gallbladder, bone marrow, adrenal gland, skin, muscle, lung, digestive tract, blood vessel, heart) or Any receptor derived from cells or the like may be used as long as it contains the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3. That is, as an apelin receptor, SEQ ID NO: 3
In addition to the protein containing the amino acid sequence represented by
Examples include a protein containing an amino acid sequence having 99.9% homology and having substantially the same activity as the protein consisting of the amino acid sequence represented by SEQ ID NO: 3.
Examples of the activity exhibited by these proteins include ligand binding activity and signal transduction. "Substantially the same quality" means that the ligand binding activity and the like are qualitatively the same. Therefore, strength and weakness such as strength of ligand binding activity and quantitative factors such as molecular weight of receptor protein may be different. Furthermore, the apelin receptor contains an N-terminal
Met protected with a protecting group (eg, formyl group, C _1-6 acyl group such as acetyl group, etc.), Gln
N-terminal side of which is cleaved in vivo and the Gln is pyroglutamine-oxidized, and the side chain of the amino acid in the molecule has an appropriate protecting group (for example, C _1-6 acyl group such as formyl group and acetyl group). It also includes protected proteins or complex proteins such as so-called glycoproteins to which sugar chains are bound. Examples of the salt of the apelin receptor include the same salts as the above-mentioned salts of apelin.

As a partial peptide of the apelin receptor,
For example, a portion of the apelin receptor molecule that is exposed outside the cell membrane is used. That is, it is a peptide containing a portion analyzed as an extracellular region (hydrophilic site) in the hydrophobicity plot analysis of the apelin receptor. Further, a peptide partially containing a hydrophobic (Hydrophobic) site can also be used. A peptide containing each domain individually may be used, but a peptide of a part containing a plurality of domains at the same time may be used. As the salt of the partial peptide of the apelin receptor, the same salts as the above-mentioned salts of apelin are used.

As the DNA encoding the apelin receptor, any DNA can be used as long as it contains a nucleotide sequence encoding the apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence of SEQ ID NO: 3. May be It may be any of genomic DNA, genomic DNA library, tissue / cell-derived cDNA, tissue / cell-derived cDNA library, and synthetic DNA. The vector used for the library may be any of bacteriophage, plasmid, cosmid, phagemid and the like. Alternatively, an RNA fraction prepared from tissues / cells can be used to directly amplify by an RT-PCR method known per se. Specifically, SEQ ID NO: 3
D encoding the apelin receptor consisting of the amino acid sequence of
As NA, DNA having the nucleotide sequence represented by SEQ ID NO: 4 or the like is used. Apelin receptors, partial peptides thereof or salts thereof, and DNAs encoding apelin receptors or partial peptides thereof are described in WO00 / 1.
It can be manufactured and adjusted by the method described in Japanese Patent No. 8793.

As the antibody against apelin or its modified form, those described in WO00 / 18793 can be used, and can be produced according to the description in WO00 / 18793. An antisense polynucleotide for apelin-encoding DNA is a polynucleotide containing a part of the base sequence of apelin-encoding DNA or a part of the base sequence complementary to the DNA, and encodes apelin. It is used not only to include DNA but also to include RNA.
An antisense polynucleotide (nucleic acid) capable of inhibiting the replication or expression of the apelin gene, a cloned or determined Apelin encoding D
It can be designed and synthesized based on the nucleotide sequence information of NA. Such a polynucleotide (nucleic acid) is the R of the apelin gene.
It can hybridize with NA, can inhibit the synthesis or function of the RNA, or can regulate and control the expression of the apelin gene through interaction with apelin-related RNA. Apelin-related RNA
Polynucleotides complementary to selected sequences of A. and polynucleotides capable of specifically hybridizing to apelin-related RNA are useful for regulating and controlling the expression of the apelin gene in vivo and in vitro. ,
It is also useful for treatment or diagnosis of diseases. The term "corresponding" means having homology or being complementary to a specific sequence of nucleotides, base sequences or nucleic acids including genes. “Corresponding” between a nucleotide (base sequence or nucleic acid) and a peptide (protein) usually refers to an amino acid of a peptide (protein) in a command derived from the nucleotide (nucleic acid) sequence or its complement. . 5'-end hairpin loop of apelin gene, 5'-end 6-base pair repeat, 5'-end untranslated region, peptide translation initiation codon, protein coding region, OR
The F translation stop codon, the 3'-end untranslated region, the 3'-end palindromic region, and the 3'-end hairpin loop can be selected as the preferred region of interest, but any region within the apelin gene can be selected as the subject.

The relationship between the target nucleic acid and the polynucleotide which is complementary to at least a part of the target region and can hybridize can be said to be "antisense" with the target. Antisense polynucleotides
Polydeoxyribonucleotides containing 2-deoxy-D-ribose, polyribonucleotides containing D-ribose, N- of purine or pyrimidine bases
Other types of polynucleotides that are glycosides,
Alternatively, other polymers with non-nucleotide backbones (eg, commercially available protein nucleic acids and synthetic sequence-specific nucleic acid polymers) or other polymers containing special linkages, provided that such polymers are found in DNA or RNA. Containing a nucleotide having a configuration that allows pairing of bases and attachment of bases) and the like. They are double-stranded DNA, single-stranded DNA, double-stranded RNA,
It may be a single-stranded RNA, further a DNA: RNA hybrid, and further has an unmodified polynucleotide (or an unmodified oligonucleotide) and an addition of a known modification, such as a label known in the art. , Capped, methylated, substituted with one or more natural nucleotides by analogs, modified intramolecularly with nucleotides such as uncharged bonds (eg methylphosphonate, phosphotriene) ester,
Those with phosphoramidates, carbamates, etc.,
Those having a charged bond or a sulfur-containing bond (eg, phosphorothioate, phosphorodithioate, etc.), such as a protein (nuclease, nuclease inhibitor, toxin, antibody, signal peptide, poly-
L-lysine, etc.), sugars (eg, monosaccharides, etc.) having side chain groups, intercurrent compounds (eg, acridine, psoralen, etc.), chelate compounds (eg, metals, radioactivity) Metal, a boron, an oxidative metal, etc.), an alkylating agent, or a modified bond (eg, α-anomeric nucleic acid).
Here, the term “nucleoside”, “nucleotide” and “nucleic acid” may include those having not only purine and pyrimidine bases but also other modified heterocyclic bases. Such modifications may include methylated purines and pyrimidines, acylated purines and pyrimidines, or other heterocycles. The modified nucleotide and the modified nucleotide may also have a modified sugar moiety, for example, one or more hydroxyl groups are substituted with halogen, an aliphatic group or the like, or a functional group such as ether or amine is added. It may have been converted.

Antisense Polynucleotide (Nucleic Acid)
Is RNA, DNA, or modified nucleic acid (RN
A, DNA). Specific examples of the modified nucleic acid include, but are not limited to, sulfur derivatives and thiophosphate derivatives of nucleic acids, and those resistant to degradation of polynucleoside amides and oligonucleoside amides. The antisense nucleic acid of the present invention can be preferably designed according to the following policy. That is, it makes the antisense nucleic acid more stable in the cell, increases the cell permeability of the antisense nucleic acid, has a greater affinity for the target sense strand, and if it is toxic, Make sense nucleic acid less toxic. Thus many modifications are known in the art, for example J. Kawakami et al., Pharm Tech Japan, Vol.
pp.247, 1992; Vol. 8, pp.395, 1992; ST Crooke e
t al. ed., Antisense Research and Applications, CR
It is disclosed in C Press, 1993, etc. Antisense nucleic acids may contain altered or modified sugars, bases, linkages, provided in specialized forms such as liposomes, microspheres, applied by gene therapy or added. It can be given in form.
Thus, as an additive form, a polycationic substance such as polylysine that acts to neutralize the charge of the phosphate skeleton, a lipid that enhances the interaction with the cell membrane or the uptake of nucleic acid ( Examples thereof include crude aqueous substances such as phospholipid and cholesterol). Preferred lipids to add include cholesterol and its derivatives (eg cholesteryl chloroformate, cholic acid, etc.). These things are
Can be attached to the 3'or 5'end of the nucleic acid,
It may be attached via a base, a sugar or an intramolecular nucleoside bond. Other groups include cap groups specifically arranged at the 3 ′ end or 5 ′ end of nucleic acid,
Examples thereof include those for preventing decomposition by nucleases such as exonuclease and RNase. Examples of the group for such a cap include protective groups for hydroxyl groups known in the art including glycols such as polyethylene glycol and tetraethylene glycol.
It is not limited to that. The inhibitory activity of antisense nucleic acid is obtained by transforming a transformant transformed with a recombinant vector containing DNA encoding apelin, a gene expression system of apelin in vivo or in vitro, or in vivo or in vitro of apelin. Can be examined using the translation system of. The nucleic acid can be applied to cells by various methods known per se.

The following are apelin or its modified form, DNA encoding apelin, antibody against apelin or modified form, anti-D against DNA encoding apelin.
Uses such as NA will be specifically described. (1) Apelin or a modified form thereof, a pharmaceutical drug containing apelin-encoding DNA or apelin receptor-encoding DNA apelin or a modified form thereof has an adrenocorticotropic hormone (ACTH) secretion-regulating action. Therefore,
When apelin or its modified form has an adrenocorticotropic hormone secretagogue action, apelin or its modified form,
Apelin-encoding DNA or apelin-receptor-encoding DNA is, for example, hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease (eg, fatigue, nausea, pigmentation, sexual function). Decrease, hair loss, hypotension), adrenal insufficiency, pain, obesity and the like. On the other hand, when apelin or its modified form has an adrenocorticotropic hormone secretion inhibitory action, apelin or its modified form, DNA coding for apelin or DNA coding for the apelin receptor is, for example, Cushing's disease, Cushing's syndrome (example , Central obesity, full moon face, hypertension, reddish purple striatum, hirsutism, adrenocorticotropic hormone (ACTH) producing pituitary tumor, CRH producing tumor, aldosterone producing tumor, aldosteronism (primary, secondary), It can be used as a prophylactic / therapeutic agent for primary cortisol resistance, congenital adrenal enzyme deficiency, Addison's disease, stress, depression, anorexia nervosa, gastroduodenal ulcer, irritable bowel syndrome and the like.

Furthermore, apelin or its modified form has an oxytocin secretion regulatory action. Therefore, when apelin or its modified form has an oxytocin secretion-promoting action, apelin or its modified form, DNA coding for apelin, or DN coding for the apelin receptor
A is, for example, a parturition-inducing agent, weak labor, relaxation bleeding, uterine restoration failure, lactation insufficiency, post-traumatic stress syndrome, milk stasis, etc. It can be used as an adjunct in cases such as abortion. On the other hand, when apelin or a modified form thereof has an oxytocin secretion inhibitory action, apelin or a modified form thereof, DNA encoding apelin or DNA encoding an apelin receptor is, for example, excessive labor pain, ankylosing uterine contraction, fetus Asphyxia, uterine rupture, cervical laceration,
It can be used as a prophylactic / therapeutic agent for dysmenorrhea, preterm birth, Prader-Willi syndrome and the like.

Apelin or its modified form, DNA encoding apelin or DN encoding apelin receptor
When A is used as the above-mentioned medicine, it can be carried out according to a conventional method. For example, tablets coated with sugar or an enteric coating as required, orally as a capsule, elixir, microcapsule, or aseptic solution with water or other pharmaceutically acceptable liquid,
Alternatively, it can be used parenterally in the form of injection such as suspension. For example, the compound or salt thereof is admixed with a physiologically acceptable carrier, flavoring agent, excipient, vehicle, preservative, stabilizer, binder and the like in a unit dosage form generally required for pharmaceutical practice. It can be manufactured by The amount of active ingredient in these preparations is such that a suitable amount within the indicated range can be obtained. The DNA encoding apelin is (a) the DNA encoding apelin or the D encoding an apelin receptor.
By administering NA to the patient and expressing it, or (b) inserting apelin-encoding DNA or apelin receptor-encoding DNA into a cell or the like and expressing it, and then transplanting the cell into the patient, etc. Can increase the amount of apelin or apelin receptor in the cells of the patient, and can fully exert the action of apelin. When a DNA encoding apelin or a DNA encoding an apelin receptor is used, the DNA is inserted alone or into an appropriate vector such as a retrovirus vector, an adenovirus vector, an adenovirus associated virus vector, and then according to a conventional method. It can be carried out.

Additives that can be mixed with tablets, capsules and the like include, for example, gelatin, corn starch, tragacanth gum, binders such as gum arabic, excipients such as crystalline cellulose, corn starch, gelatin, alginic acid and the like. Such as a leavening agent, a lubricant such as magnesium stearate, a sweetener such as sucrose, lactose or saccharin, and a flavoring agent such as peppermint, red oil or cherry. When the unit dosage form is a capsule, a liquid carrier such as fat may be included in addition to materials of the above type. Sterile compositions for injection can be formulated according to conventional pharmaceutical practices such as dissolving or suspending the active substance in a vehicle such as water for injection, naturally occurring vegetable oils such as sesame oil, coconut oil, etc. .. Examples of the aqueous solution for injection include physiological saline, isotonic solution containing glucose and other adjuvants (eg, D-sorbitol, D-mannitol, sodium chloride, etc.), and suitable solubilizing agents. , Eg alcohols (eg ethanol), polyalcohols (eg propylene glycol, polyethylene glycol), nonionic surfactants (eg polysorbate 80 (TM), HCO-5).
0) etc. may be used together. Examples of the oily liquid include sesame oil and soybean oil, which may be used in combination with solubilizing agents such as benzyl benzoate and benzyl alcohol. Also,
Buffer (eg, phosphate buffer, sodium acetate buffer), soothing agent (eg, benzalkonium chloride, procaine hydrochloride, etc.), stabilizer (eg, human serum albumin, polyethylene glycol, etc.), preservative ( For example,
(Benzyl alcohol, phenol, etc.), antioxidant, etc. The prepared injection solution is usually filled in a suitable ampoule. Since the preparation thus obtained is safe and has low toxicity, for example, humans and mammals (for example, mouse, rat, guinea pig, rabbit, chicken,
Sheep, pigs, cows, cats, dogs, monkeys, baboons,
Chimpanzee). The dose of apelin, apelin-encoding DNA or apelin-receptor-encoding DNA varies depending on the symptoms, etc., but oral administration generally results in adult hypoaldosteronism (weight 60 kg). About 0.1 to 100 mg per day, preferably about 1.0 to
50 mg, more preferably about 1.0-20 mg.
In the case of parenteral administration, the single dose varies depending on the administration subject, target organ, condition, administration method, etc., but for example in the form of an injection, it can be administered to an adult patient with hypoaldosteronism (weight 60 kg). In administration, about 0.01 to 30 mg per day, preferably about 0.1
It is convenient to administer about -20 mg, more preferably about 0.1-10 mg by intravenous injection. In the case of other animals, the dose converted per 60 kg can be administered.

(2) Pharmaceuticals Containing Antibodies to Apelin Antibodies having the activity of neutralizing the activity of apelin can suppress the adrenocorticotropin (ACTH) secretion promoting activity of apelin. (A
CTH) secretion inhibitors include, for example, Cushing's disease, Cushing's syndrome (eg, central obesity, full-face like face, hypertension, purple-red skin striae, hirsutism), adrenocorticotropic hormone (A
CTH) producing pituitary tumor, CRH producing tumor, aldosterone producing tumor, aldosteronism (primary and secondary), primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa , A gastric / duodenal ulcer, an irritable bowel syndrome, etc. can be used as a medicine for preventing / treating agents. Furthermore, since an antibody having an action of neutralizing the activity of apelin can suppress the oxytocin secretion promoting action of apelin, as an oxytocin secretion inhibitor, for example, excessive labor pain, ankylosing uterine contraction, fetal asphyxia, uterus Rupture, cervical laceration, dysmenorrhea,
It can be used as a medicine such as a prophylactic / therapeutic agent for preterm birth and Prader-Willi syndrome. The therapeutic / prophylactic agent for the above-mentioned diseases containing the antibody of the present invention, as a liquid formulation as it is, or as a pharmaceutical composition in an appropriate dosage form, human or mammal (eg, rat, rabbit, sheep, pig, cow, cat,
Can be orally or parenterally administered to dogs, monkeys, etc.). The dose varies depending on the administration subject, target disease, symptom, administration route, etc., but when it is used for an adult, for example, the antibody of the present invention is usually administered in a dose of about 0.01 to 20 mg / kg body weight. , Preferably 0.1
-10 mg / kg body weight, more preferably 0.1-
It is convenient to administer about 5 mg / kg body weight by intravenous injection about 1 to 5 times a day, preferably about 1 to 3 times a day. In the case of other parenteral administration and oral administration, an amount similar to this can be administered. If the symptoms are particularly severe, the dose may be increased according to the symptoms.

The antibody of the present invention can be administered per se or as an appropriate pharmaceutical composition. The pharmaceutical composition used for the above administration contains the above or a salt thereof and a pharmacologically acceptable carrier, diluent or excipient. Such a composition is provided as a dosage form suitable for oral or parenteral administration. That is, for example, as a composition for oral administration, solid or liquid dosage forms, specifically tablets (including sugar-coated tablets, film-coated tablets),
Examples include pills, granules, powders, capsules (including soft capsules), syrups, emulsions and suspensions.
Such a composition is produced by a method known per se and contains a carrier, a diluent or an excipient which is usually used in the field of formulation. For example, lactose, starch, sucrose, magnesium stearate and the like are used as carriers and excipients for tablets. As the composition for parenteral administration, for example, injections, suppositories, etc. are used, and the injections are in the form of intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, drip injection, etc. Includes. Such an injection is prepared according to a method known per se, for example, by dissolving, suspending or emulsifying the above-mentioned antibody or a salt thereof in a sterile aqueous or oily liquid usually used for injection. As the aqueous solution for injection, for example, physiological saline, isotonic solution containing glucose and other auxiliary agents, and the like,
Suitable solubilizing agents such as alcohol (eg ethanol), polyalcohol (eg propylene glycol, polyethylene glycol), nonionic surfactant [eg polysorbate 80, HCO-50 (polyoxyethylene).
(50 mol) adduct of hydrogenated castor oil)] and the like. As the oily liquid, for example, sesame oil, soybean oil and the like are used, and solubilizing agents such as benzyl benzoate and benzyl alcohol may be used in combination.
The prepared injection solution is usually filled in a suitable ampoule. The suppository used for rectal administration is prepared by mixing the above antibody or a salt thereof with an ordinary suppository base. The oral or parenteral pharmaceutical compositions described above are conveniently prepared in dosage unit form for administration of the active ingredient. Examples of the dosage form of such a dosage unit include tablets, pills, capsules, injections (ampoules), suppositories, etc., usually 5 to 500 mg per dosage unit dosage form, especially 5 to 100 mg for injections,
Other dosage forms preferably contain 10-250 mg of the antibody. Each composition described above may contain other active ingredients as long as the composition does not cause an unfavorable interaction with the antibody.

(3) Diagnostic agent containing antibody against apelin Since an antibody against apelin can specifically recognize apelin, it is used for quantification of apelin in a test solution, especially for quantification by sandwich immunoassay. can do. The method for quantifying apelin using an antibody against apelin can be carried out according to the method described in WO99 / 33976. Therefore, an antibody against apelin is abnormally expressed (overexpression, decreased expression)
Secretion of adrenocorticotropic hormone (ACTH) associated with
Abnormal secretion of oxytocin can be detected. Specifically, the antibody against apelin is the secretion of adrenocorticotropic hormone (ACTH) associated with aberrant expression of apelin in humans or mammals (eg, rat, rabbit, sheep, pig, cow, cat, dog, monkey, etc.). Abnormal aldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease (eg, fatigue, nausea, pigmentation, hypogonadism, hair loss, hypotension), adrenal insufficiency,
Pain, obesity, Cushing's disease, Cushing's syndrome (eg, central obesity, full moon face, hypertension, reddish purple striatum, hirsutism), adrenocorticotropic hormone (ACTH) -producing pituitary tumor, C
RH-producing tumor, aldosterone-producing tumor, aldosteronism (primary, secondary), primary cortisol resistance,
It can be used as a diagnostic agent for congenital adrenal enzyme deficiency, Addison's disease, stress, depression, anorexia nervosa, gastroduodenal ulcer, irritable bowel syndrome and the like. Also,
Antibodies against apelin, weak labor due to abnormal secretion of oxytocin associated with abnormal expression of apelin in humans or mammals, laxative bleeding, uterine restoration failure, lactation failure,
It can be used as a diagnostic agent for post-traumatic stress syndrome, milk stasis, excessive labor pain, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth, Prader-Willi syndrome and the like.

(4) Genetic Diagnostic Agent A DNA encoding apelin can be used, for example, as a probe to obtain human or mammalian (eg,
Rat, mouse, guinea pig, rabbit, bird, sheep,
Since it is possible to detect an abnormality (gene abnormality) of DNA or mRNA encoding apelin in pig, cow, horse, cat, dog, monkey, etc., for example, the DN
It is useful as a gene diagnostic agent for damage, mutation or decreased expression of A or mRNA, and increased or excessive expression of the DNA or mRNA. The above-mentioned gene diagnosis using a DNA encoding apelin or an antisense polynucleotide thereto can be carried out by, for example, Northern hybridization or PCR-SSC known per se.
P method (Genomics, Volume 5, 874-8
79 pages (1989), Proceedings of the
Proceedings of the National Academ
y of Sciences of the United States of America),
Vol. 86, pp. 2766-2770 (1989)) and the like. For example, when abnormal expression of apelin is detected by Northern hybridization, hypoaldosteronism, hypocortisolemia, secondary and chronic adrenocortical dysfunction due to abnormal secretion of adrenocorticotropic hormone (ACTH), Addison's disease ( Example,
Fatigue, nausea, pigmentation, hypogonadism, hair loss, hypotension),
Adrenal insufficiency, pain, obesity, Cushing's disease, Cushing's syndrome (eg, central obesity, full face, hypertension, reddish violet striatum, hirsutism), corticotropin (ACTH) -producing pituitary tumor, CRH-producing tumor , Aldosterone-producing tumors,
Aldosteronism (primary, secondary), primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa, gastroduodenal ulcer,
It can be diagnosed as being likely to have a disease such as irritable bowel syndrome or likely to be affected in the future. In addition, when abnormal expression of apelin is detected by Northern hybridization, weak labor, relaxation bleeding due to abnormal secretion of oxytocin accompanying abnormal expression of apelin,
Uterine retrograde failure, lactation failure, post-traumatic stress syndrome,
Diseases such as milk stasis, overactive labor, ankylosing contractions, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth, Prader-Willi syndrome, etc. It can be diagnosed as high.

(5) DN containing antisense DNA
Antisense DN to DNA encoding A apelin
A can suppress the adrenocorticotropic hormone (ACTH) secretagogue activity of apelin, and therefore, as an adrenocorticotropic hormone (ACTH) secretion inhibitor, for example, Cushing's disease, Cushing's syndrome (eg, central obesity, full moon) Facial appearance, hypertension, reddish purple striatum, hirsutism, adrenocorticotropic hormone (ACTH) producing pituitary tumor, CRH producing tumor, aldosterone producing tumor, aldosteronism (primary, secondary), primary cortisol resistance, It can be used as a medicine such as a prophylactic / therapeutic agent for congenital adrenal enzyme deficiency, Addison's disease, stress, depression, anorexia nervosa, gastric / duodenal ulcer, irritable bowel syndrome and the like. The antisense DNA against DNA encoding apelin can suppress the oxytocin secretion promoting action of apelin, and therefore, as an oxytocin secretion inhibitor, for example, for example, excessive labor pain, ankylosing uterine contraction, fetal distress, uterine rupture, It can be used as a medicine such as a prophylactic / therapeutic agent for cervical laceration, dysmenorrhea, preterm birth, Prader-Willi syndrome and the like. Antisense DNA against apelin-encoding DNA is apelin adrenocorticotropic hormone (ACT).
H) Since it can suppress the secretagogue activity, it can be used as an adrenocorticotropic hormone (ACTH) secretion inhibitor, for example, Cushing's disease, Cushing's syndrome (eg, central obesity, full face like face, hypertension, reddish purple skin striae). , Hirsutism), adrenocorticotropic hormone (ACTH) -producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism (primary and secondary), primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress , A depression, anorexia nervosa, gastric / duodenal ulcer, irritable bowel syndrome, etc. can be used as a medicine for preventing / treating agents.
Furthermore, since antisense DNA against DNA encoding apelin can suppress the oxytocin secretion promoting action of apelin, it can be used as an oxytocin secretion inhibitor, for example, as hypertonic labor, ankylosing uterine contraction, fetal asphyxia,
Uterine rupture, cervical laceration, dysmenorrhea, preterm birth, Prader-Willi
It can be used as a medicine such as a prophylactic / therapeutic agent for syndrome and the like. When the antisense DNA is used as the therapeutic / prophylactic agent, the antisense DNA is
It can be formulated in the same manner as the DNA encoding apelin. The preparation thus obtained has low toxicity and should be orally or parenterally administered to humans or mammals (eg, rat, rabbit, sheep, pig, cow, cat, dog, monkey, etc.) You can The antisense DNA can be administered as it is or with a physiologically acceptable carrier such as an auxiliary agent for promoting ingestion through a gene gun or a catheter such as a hydrogel catheter. The dose of the antisense DNA varies depending on the target disease, administration subject, administration route, etc., but for example, for the purpose of treating Cushing's disease, the antisense DNA is used as an organ (eg, liver, lung, heart, kidney, etc.).
When administered topically to an adult (body weight 60 kg),
It is about 0.1 to 100 mg per day.

Double-stranded RNA containing a part of RNA encoding apelin and RNA complementary thereto
(RNAi; RNA interference method), ribozymes containing a part of apelin-encoding RNA, and the like can suppress the expression of apelin-encoding DNA in the same manner as the above-mentioned antisense DNA, and apelin in vivo or The function of DNA encoding apelin can be suppressed. Therefore, since the double-stranded RNA or ribozyme can suppress the adrenocorticotropic hormone (ACTH) secretion promoting activity of apelin, it can be used as an adrenocorticotropic hormone (ACTH) secretion inhibitor, for example, Cushing's disease, Cushing's syndrome. (Eg, central obesity, full moon face, hypertension, reddish purple striatum, hirsutism), adrenocorticotropic hormone (ACTH) producing pituitary tumor, CRH producing tumor, aldosterone producing tumor, aldosteronism (primary, secondary ), Primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa, gastric / duodenal ulcer, irritable bowel syndrome, etc. You can further,
The double-stranded RNA or ribozyme can suppress the oxytocin secretion-promoting action of apelin, and therefore, as an oxytocin secretion inhibitor, for example, overwork pain, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, It can be used as a drug such as a prophylactic / therapeutic agent for dysmenorrhea, preterm birth, Prader-Willi syndrome, and the like. Double-stranded RNA is
It can be designed and manufactured based on the sequence of the DNA of the present invention according to a known method (eg, Nature, 411, 494, 2001). Ribozymes can be prepared by known methods (eg TRENDS i
n Molecular Medicine, Vol. 7, p. 221, 2001), and can be designed and manufactured based on the apelin DNA sequence. For example, it can be produced by linking a known ribozyme to a part of RNA encoding apetin. As a part of RNA encoding apelin, a part (RNA fragment) close to the cleavage site on RNA of apelin that can be cleaved by a known ribozyme can be mentioned. When the above-mentioned double-stranded RNA or ribozyme is used as the above-mentioned preventive / therapeutic agent, antisense DN
It can be formulated and administered in the same manner as in A.

(6) Screening Method The screening method of the present invention is characterized by using (6A-1) apelin or a modified form thereof and / or apelin receptor (hereinafter, also referred to as partial peptides) for adrenal cortex stimulation. Method for screening hormone secretion regulator, (6A-2) DNA containing DNA encoding apelin and / or DNA encoding apelin receptor are used for screening adrenocorticotropic hormone secretion regulator , (6B-
1) A screening method for an oxytocin secretion regulator, which comprises using apelin or a modified form thereof and / or an apelin receptor, and (6B-2)
A method for screening an oxytocin secretion regulator, which comprises using a DNA containing a DNA encoding apelin and / or a DNA encoding an apelin receptor. Corticotropin and oxytocin secretion regulators include substances that change the binding properties of apelin or its modified form to apelin receptors, or substances that regulate the expression of apelin and / or apelin receptors. Can be mentioned.

First, a method for screening a substance that changes the binding property between apelin and the apelin receptor will be described. By using an apelin receptor or constructing a recombinant apelin receptor expression system and using a receptor binding assay system using the expression system, a substance that changes the binding property between apelin and an apelin receptor can be identified. Can be screened. Such substances include substances having cell stimulating activity via apelin receptors (that is, apelin receptor agonists), substances having no such cell stimulating activity (that is, apelin receptor antagonists), and the like. The term “alter the binding property between apelin and apelin receptor” includes both the case of inhibiting the binding between apelin and apelin receptor and the case of promoting the binding between apelin and apelin receptor. is there.
Examples of the cell stimulating activity include arachidonic acid release, acetylcholine release, intracellular Ca ²⁺ release, intracellular cAM.
P production, intracellular cGMP production, inositol phosphate production, cell membrane potential fluctuation, intracellular protein phosphorylation, cf
Examples of the activity include activating os and promoting or suppressing pH decrease. Among them, intracellular cAM
P inhibitory activity is preferred.

That is, the present invention relates to (i) the case where apelin or its modified form is brought into contact with the apelin receptor, and (i)
i) A method for screening a substance that alters the binding property between apelin and an apelin receptor, which is characterized by performing a comparison with the case where the apelin receptor is contacted with apelin and a test compound. In the screening method of the present invention, for example, the binding amount of apelin to the apelin receptor in (i) the case where apelin is brought into contact with apelin and (ii) the case where apelin and the test compound are brought into contact with the apelin receptor, Cell stimulating activity etc. are measured and compared.

Specifically, the screening method of the present invention comprises contacting labeled apelin or a modified form thereof with an apelin receptor and contacting labeled apelin or a modified form thereof and a test compound with an apelin receptor. In the case of, a method for screening a substance that changes the binding property between apelin and an apelin receptor, which comprises measuring and comparing the amount of binding of the labeled apelin or its modified form to the apelin receptor, labeled apelin or When the modified product is brought into contact with a cell containing an apelin receptor or a membrane fraction of the cell, the labeled apelin or a modified product thereof and a test compound are added to a cell containing the apelin receptor or a membrane fraction of the cell. The labeled apelin or its modified form to the cell or the membrane fraction when contacted with A method for screening a substance that changes the binding property between apelin and an apelin receptor, characterized by measuring and comparing the amount of binding, a labeled apelin or a modified form thereof containing a DNA encoding an apelin receptor When a transformant is contacted with an apelin receptor expressed on the cell membrane by culturing the transformant, a labeled apelin or a modified form thereof and a test compound are cultured in a transformant containing a DNA encoding the apelin receptor. When the apelin receptor expressed on the cell membrane is brought into contact with the apelin receptor by measuring the binding amount of the labeled apelin or its modified product to the apelin receptor, and comparing the apelin and the apelin receptor. A method for screening a substance that changes the binding property,

When a compound that activates the apelin receptor (eg, apelin or a modified form thereof) is contacted with a cell containing the apelin receptor, a compound that activates the apelin receptor and a test compound are added to the apelin receptor. Of a substance which changes the binding property between apelin and an apelin receptor, which comprises measuring and comparing the cell stimulating activity mediated by the apelin receptor when brought into contact with cells containing apelin, and apelin A compound that activates the receptor (for example, apelin or a modified form thereof) that encodes the apelin receptor
When a transformant containing NA is brought into contact with an apelin receptor expressed on the cell membrane by culturing, a compound containing a DNA encoding the apelin receptor is used as a compound activating the apelin receptor and a test compound. Binding of apelin and apelin receptor characterized by measuring and comparing cell stimulating activity mediated by apelin receptor when contacted with apelin receptor expressed on cell membrane by culturing transformant For screening a substance that changes the apelin receptor, and measuring and comparing the cell stimulating activity mediated by the apelin receptor when a test compound is contacted with cells containing the apelin receptor. Screening method, transformant containing test compound containing DNA encoding apelin receptor In the case of contacting the apelin receptor expressed on the cell membrane by culturing, and measuring the cell stimulating activities mediated by the apelin receptor screening method of apelin receptor agonist and comparing the, or the like. In the screening method of the present invention, instead of using apelin as a ligand, a compound that changes the binding property between apelin and an apelin receptor (for example, a low molecular synthetic compound, preferably a low molecular synthetic agonist) or a salt thereof Can also be used. The compound or its salt that changes the binding property between apelin and the apelin receptor can be obtained, for example, by carrying out the screening method of the present invention using apelin as a ligand. Specifically, the above-described modified apelin and the like can be used.

A detailed description of the screening method of the present invention is given below. First, the apelin receptor used in the screening method of the present invention may be any as long as it contains the above-mentioned apelin receptor, but it may be a membrane fraction of an organ of human or warm-blooded animal. It is suitable.
However, since human-derived organs are extremely difficult to obtain, apelin receptors expressed in large amounts using recombinants and the like are suitable for use in screening. In the screening method of the present invention, when a cell containing an apelin receptor or a cell membrane fraction thereof is used, the preparation method described below may be followed. When a cell containing an apelin receptor is used, the cell may be fixed with glutaraldehyde, formalin or the like. The immobilization method can be performed according to a method known per se. The cell containing the apelin receptor refers to a host cell expressing the apelin receptor, and the host cell includes the above-mentioned Escherichia coli, Bacillus subtilis, yeast, insect cell, animal cell and the like. The membrane fraction refers to a fraction containing a large amount of cell membrane obtained by a method known per se after crushing cells. As a method of crushing cells, a method of crushing cells with a Potter-Elvehjem type homogenizer, crushing with a Waring blender or polytron (Kinematica), crushing with ultrasonic waves, and blasting cells from a thin nozzle while applying pressure with a French press etc. Examples include crushing by things. For the fractionation of the cell membrane, a fractionation method by centrifugal force such as a fractionation centrifugation method or a density gradient centrifugation method is mainly used. For example, the cell lysate may be fed at a low speed (500 rpm to 3
Centrifuge at 000 rpm for a short time (usually about 1 to 10 minutes), and then the supernatant is spun at a higher speed (15,000 to 30,000 rpm).
It is usually centrifuged for 30 minutes to 2 hours at (rpm) and the resulting precipitate is used as a membrane fraction. The membrane fraction is rich in expressed apelin receptors and membrane components such as cell-derived phospholipids and membrane proteins. The amount of apelin receptor in cells or membrane fraction containing the apelin receptor is 10 ³ to 10 ⁸ per cell.
It is preferably a molecule, and is preferably 10 ⁵ to 10 ⁷ molecules. It should be noted that the higher the expression level, the higher the ligand binding activity (specific activity) per membrane fraction, making it possible not only to construct a highly sensitive screening system, but also to measure a large number of samples in the same lot. .

In order to carry out the above steps (1) to (3) for screening a substance that alters the binding property between apelin and an apelin receptor, a suitable apelin receptor fraction and a labeled apelin or a modified form thereof are used. The apelin receptor fraction is preferably a natural apelin receptor fraction or a recombinant apelin receptor fraction having an activity equivalent to that. Here, the equivalent activity refers to equivalent ligand binding activity and the like. As the labeled apelin or its modified product, a labeled ligand, a labeled ligand analog compound, or the like is used. For example, [3H],
^[125 I], ^{[1 4} C], it can be utilized such as apelin or modification thereof that is labeled with a ^[35 S]. Specifically, in order to screen a substance that changes the binding property between apelin or a modified form of apelin and the apelin receptor, first, a cell containing the apelin receptor or a membrane fraction of the cell is treated with a buffer suitable for screening. A receptor preparation is prepared by suspending in. Any buffer may be used as long as it does not inhibit the binding between apelin and an apelin receptor, such as a phosphate buffer having a pH of 4 to 10 (preferably a pH of 6 to 8) and a Tris-hydrochloric acid buffer. Further, for the purpose of reducing non-specific binding, a surfactant such as CHAPS, Tween-80 ^™ (Kao-Atlas), digitonin and deoxycholate may be added to the buffer. Furthermore, PMSF, leupeptin, for the purpose of suppressing the degradation of apelin receptor, apelin or its modified product by proteases,
A protease inhibitor such as E-64 (produced by Peptide Laboratories) or pepstatin can also be added. 0.01m
1 to 10 ml of the apelin receptor solution, a fixed amount (50
It was added labeled apelin or modifications thereof of 00cpm~500000cpm), at the same time ^{10 -} 4 ^{-10 -1}
Co-exist with μM test compound. Non-specific binding (NS
To know B), prepare a reaction tube containing a large excess of unlabeled apelin or its modified product. The reaction is about 0 ℃
To about 50 ° C., preferably about 4 ° C. to about 37 ° C. for about 20 minutes
It is performed for about 24 hours, preferably for about 30 minutes to about 3 hours. After the reaction, the mixture is filtered through a glass fiber filter paper or the like, washed with an appropriate amount of the same buffer, and the radioactivity remaining on the glass fiber filter paper is measured with a liquid scintillation counter or a γ-counter. Count (B ₀ −) obtained by subtracting the amount of non-specific binding (NSB) from the count (B ₀ ) in the absence of an antagonist.
NSB) as 100%, the specific binding amount (B-NS
A test compound having B) of, for example, 50% or less can be selected as a candidate substance capable of competitive inhibition.

In order to carry out the above methods (1) to (3) for screening a substance that changes the binding property between apelin and an apelin receptor, a known method or a commercially available kit for measuring the cell stimulating activity mediated by the apelin receptor is used. Can be used to measure. Specifically, first, cells containing the apelin receptor are cultured in a multiwell plate or the like. For screening, the medium was exchanged with a fresh medium or an appropriate buffer that did not show toxicity to cells in advance, and after adding a test compound or the like and incubating for a certain period of time, cells were extracted or supernatant was collected to generate The product is quantified according to the respective method. Substances used as indicators of cell-stimulating activity (eg arachidonic acid, cA
When it is difficult to assay the production of MP, etc. by the degrading enzyme contained in the cells, an inhibitor may be added to the degrading enzyme to perform the assay. Further, the activity of suppressing cAMP production and the like can be detected as a production suppressing action on cells in which the basic production amount of cells has been increased by forskolin or the like. In order to measure the cell stimulating activity and perform screening, cells expressing an appropriate apelin receptor are required. As the cells expressing the apelin receptor of the present invention, the above-mentioned recombinant apelin receptor-expressing cell line and the like are desirable. Examples of test compounds include peptides, proteins, non-peptidic compounds, synthetic compounds, fermentation products, cell extracts, plant extracts, animal tissue extracts, and the like, and these compounds may be novel compounds. However, it may be a known compound. Also,
As the test compound, a compound designed to bind to the ligand binding pocket based on the atomic coordinates of the active site of the apelin receptor and the position of the ligand binding pocket is preferably used. The atomic coordinates of the active site of the apelin receptor and the position of the ligand binding pocket can be measured using a known method or a method similar thereto.

A kit for screening a substance that changes the binding property between apelin and an apelin receptor comprises: Is included. Examples of the screening kit of the present invention include the following. 1. To the screening reagent measurement buffer and washing buffer Hanks' Balanced Salt Solution (manufactured by Gibco), 0.
To which 05% bovine serum albumin (manufactured by Sigma) was added. Sterilize by filtration with a 0.45μm filter at 4 ℃
It may be stored at or prepared at the time of use. Apelin receptor preparation CHO cells expressing apelin receptor were subcultured on a 12-well plate at 5 × 10 ⁵ cells / well and incubated at 37 ° C., 5% C
Cultured in O ₂ and 95% air for 2 days. Apelin labeled with labeled ligand [ ³ H], [ ¹²⁵ I], [ ¹⁴ C], [ ³⁵ S], etc. Then, dilute to 1 μM with the measurement buffer before use. The ligand standard solution apelin or its modified form is dissolved in PBS containing 0.1% bovine serum albumin (manufactured by Sigma) to 1 mM and stored at -20 ° C.

2. Assay method Apelin receptor-expressing cells cultured in a 12-well tissue culture plate are washed twice with 1 ml of measurement buffer, and then 490 μl of measurement buffer is added to each well. After adding 5 μl of a test compound solution of 10 ⁻³ to 10 ⁻¹⁰ M, 5 μl of labeled apelin or its modified product is added, and the mixture is reacted at room temperature for 1 hour. In order to know the amount of non-specific binding, 5 μl of 10 ⁻³ M ligand was added in place of the test compound. Remove the reaction solution and wash 3 times with 1 ml of wash buffer. The cell-bound labeled apelin or its modified form is lysed with 0.2N NaOH-1% SDS and mixed with 4 ml of liquid scintillator A (manufactured by Wako Pure Chemical Industries). Liquid scintillation counter (Beckman)
Radioactivity was measured using a Percent Maximum Binding
(PMB) is calculated by the following equation [Equation 1]. [Number 1] PMB = [(B- NSB) / (B 0 -NSB)] × 10
0 PMB: Percent Maximum Binding B: Value when a sample is added NSB: Non-specific Binding B ₀ : Maximum binding amount

A substance obtained by using the screening method or the screening kit of the present invention is a substance that changes (inhibits or promotes) the binding between apelin and an apelin receptor, and specifically, the apelin receptor It is a substance having a cell stimulating activity through (a so-called apelin receptor agonist) or a compound having no such stimulating activity (a so-called apelin receptor antagonist). Examples of the substance include peptides, proteins, non-peptidic compounds, synthetic compounds and fermentation products selected from the above-mentioned test compounds, and these compounds may be novel compounds or known compounds. Good. The specific evaluation method for determining whether it is an apelin receptor agonist or an antagonist may follow (i) or (ii) below. (I) After performing the binding assay shown in the above-mentioned screening methods to obtain a substance that changes the binding property of apelin or its modified form to the apelin receptor (in particular, inhibits the binding), the substance is It is determined whether or not it has the cell stimulating activity mediated by the apelin receptor described above. A substance having a cell stimulating activity is an apelin receptor agonist, and a substance having no such activity is an apelin receptor antagonist. (Ii) (a) A test compound is brought into contact with cells containing an apelin receptor, and the cell-stimulating activity mediated by the apelin receptor is measured. The substance having cell stimulating activity is an apelin receptor agonist. (b) when a compound that activates the apelin receptor (for example, apelin or a modified form thereof) is contacted with cells containing the apelin receptor, and when a compound that activates the apelin receptor and a test compound are The cell-stimulating activity mediated by the apelin receptor when contacted with cells containing the body is measured and compared. The substance capable of reducing the cell stimulating activity of the compound activating the apelin receptor is an apelin receptor antagonist. As the cell stimulating activity used as an index, for example, an inhibitory activity of intracellular cAMP production is preferable.

Since the apelin receptor agonist has the same physiological activity as that of apelin, it is useful as a safe and low-toxic drug like apelin or its modified form. On the contrary, since the apelin receptor antagonist can suppress the physiological activity of apelin, it is useful as a safe and low toxic pharmaceutical that suppresses the activity of apelin or its modified form. Since apelin has an adrenocorticotropic hormone secretion regulating action (preferably an adrenocorticotropic hormone secretagogue action) or an oxytocin secretion regulating action (preferably an oxytocin secretagogue action), the above-mentioned apelin receptor agonist or apelin The receptor antagonist also has an adrenocorticotropic hormone secretion regulating action or an oxytocin secretion regulating action. More specifically, the apelin receptor agonist has an adrenocorticotropic hormone secretagogue action or an oxytocin secretagogue action, and the apelin receptor antagonist has an adrenocorticotropic hormone secretagogue action or an oxytocin secretagogue action. Therefore, a substance that changes the binding between apelin and an apelin receptor can be used as a medicine such as an adrenocorticotropic hormone secretory regulator or an oxytocin secretory regulator. Specifically, apelin receptor agonists include, as adrenocorticotropic hormone secretagogues, for example, hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease (fatigue, nausea, pigmentation). , Hypofunction, hair loss, hypotension), adrenal insufficiency, pain, obesity, etc. In addition, the apelin receptor agonist, as an oxytocin secretagogue,
For example, labor inducement agents, weak labor, flaccid bleeding, uterine retrograde failure, lactation insufficiency, posttraumatic stress syndrome, milk stasis, etc. It can be used as a medicine such as an auxiliary agent. Apelin receptor antagonists are adrenocorticotropic hormone secretion inhibitors, for example, Cushing's disease, Cushing's syndrome (central obesity, full moon face, hypertension, purple-red skin striae, hirsutism), adrenocorticotropic hormone (ACTH) production. Pituitary tumor, CRH producing tumor,
Aldosterone-producing tumor, aldosteronism (primary,
Secondary), primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa, gastroduodenal ulcer, irritable bowel syndrome, etc.
It can be used as a medicine such as a therapeutic agent. Also,
Apelin receptor antagonists are oxytocin secretion inhibitors, for example, hypertonic labor, ankylosing uterine contractions, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth, Prader.
-It can be used as a medicine such as a prophylactic / therapeutic agent for Willi syndrome.

The substance obtained by using the above-mentioned screening method or screening kit may form a salt, for example, a pharmaceutically acceptable salt is used. Specifically, a salt with an inorganic base, a salt with an organic base,
Examples thereof include salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. Preferable examples of the salt with an inorganic base include alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, and aluminum salt and ammonium salt. Preferable examples of the salt with an organic base include, for example, trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N, N '.
-Examples include salts with dibenzylethylenediamine and the like.
Preferable examples of salts with inorganic acids include salts with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid and the like. Preferable examples of salts with organic acids include, for example, formic acid, acetic acid, propionic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, benzoic acid. And salt. Suitable examples of salts with basic amino acids include, for example, salts with arginine, lysine, ortinine, and suitable examples with acidic amino acids include, for example, aspartic acid,
Examples include salts with glutamic acid. When the compound obtained by using the screening method of the present invention is used as the above-mentioned therapeutic / prophylactic agent, it can be carried out in a conventional manner. For example, tablets, capsules, elixirs, microcapsules, sterile solutions, in the same manner as the above-mentioned pharmaceutical composition containing apelin or a modified form thereof.
It can be a suspension agent or the like. Since the preparation thus obtained is safe and has low toxicity, for example, humans or warm-blooded animals (for example, mouse, rat, rabbit, sheep, pig, cow, horse, bird, cat, dog, monkey, chimpanzee, etc.) ). The dose of the compound or its salt varies depending on its action, target disease, administration subject, administration route and the like. For example, when an apelin receptor agonist is orally administered, it is generally in an adult (per 60 kg body weight). Is about 0.1-100 mg, preferably about 1.0-, of the compound per day.
50 mg, more preferably about 1.0-20 mg is administered. When administered parenterally, the single dose of the compound varies depending on the administration subject, target disease, etc., but for example, when an apelin receptor agonist is usually administered to an adult (per 60 kg) in the form of an injection About 0.01 to 30 mg of the compound per day, preferably about 0.1 to
It is convenient to administer about 20 mg, more preferably about 0.1 to 10 mg by intravenous injection. In the case of other animals, the dose converted per 60 kg can be administered.

Next, a method for screening a substance that regulates the expression of apelin and / or apelin receptor will be described. Specifically, the screening method of the present invention comprises (1) (i) the expression level of apelin or the expression level of each apelin in the case of culturing cells or tissues capable of expressing apelin in the presence and absence of a test compound. A method for screening a substance that promotes or inhibits the expression of apelin, which comprises measuring and comparing the amount of mRNA encoding apelin, (2) (i) a cell or tissue capable of expressing an apelin receptor, and a test compound A substance that promotes or inhibits the expression of an apelin receptor characterized by measuring and comparing the expression level of each apelin receptor or the amount of mRNA encoding apelin in the presence and absence of Is a screening method. As cells or tissues capable of expressing apelin or apelin receptor, human or warm-blooded animals (eg, guinea pig, rat, mouse, chicken, rabbit, pig, sheep, cow, monkey, etc.) cells (for example, nerve cells, Endocrine cells, neuroendocrine cells, glial cells, pancreatic β cells, bone marrow cells, hepatocytes, spleen cells, mesangial cells, epidermal cells,
Epithelial cells, endothelial cells, fibroblasts, fiber cells, muscle cells,
Adipocytes, immune cells (eg macrophages, T cells, B
Cells, natural killer cells, mast cells, neutrophils, basophils, eosinophils, monocytes, dendritic cells), megakaryocytes, synovial cells,
Chondrocytes, osteocytes, osteoblasts, osteoclasts, mammary gland cells, or stromal cells, or precursors of these cells, stem cells or cancer cells, etc.) or any tissue in which these cells are present, for example, the brain, Each part of the brain (eg, olfactory bulb,
Amygdala, basal sphere, hippocampus, thalamus, hypothalamus, cerebral cortex, medulla oblongata, cerebellum), spinal cord, pituitary gland, stomach, pancreas, kidney, liver, gonad, thyroid, gallbladder, bone marrow, adrenal gland, skin, muscle, lung , Digestive tract (eg, large intestine, small intestine), blood vessel, heart, thymus, spleen, salivary gland, peripheral blood, prostate, testis (testis), ovary, placenta, uterus, bone, cartilage, joint, skeletal muscle, etc. good. At that time, cell lines and primary culture system may be used.
Further, a transformant transformed with a recombinant vector containing the above-mentioned DNA encoding apelin or apelin receptor may be used. The method for culturing cells capable of expressing apelin or the apelin receptor is the same as the above-mentioned method for culturing transformants. As the test compound, a DNA library or the like can be used in addition to the above test compounds.

The expression level of apelin or an apelin receptor can be measured by a known method such as an immunochemical method using an antibody or the like, or the mR encoding apelin.
NA for Northern hybridization, RT-PC
It can also be measured by a known method using R or TaqMan PCR method. To compare the expression levels of mRNAs by a hybridization method, a known method or a method similar thereto, for example, Molecular Cloning 2nd (J. Sambrooke
al., Cold Spring Harbor Lab. Press, 1989). Specifically, the amount of mRNA encoding apelin or apelin receptor is measured by RNA extracted from cells according to a known method.
And DNA encoding apelin or apelin receptor
Or contacting with a part thereof or the antisense polynucleotide of the present invention, and measuring the amount of the DNA encoding apelin or apelin receptor or the part thereof or the mRNA bound to the antisense polynucleotide of the present invention. Done by DNA or a part thereof encoding apelin or an apelin receptor or by labeling the antisense polynucleotide of the present invention with, for example, a radioisotope, a dye or the like, Alternatively, the amount of mRNA bound to the antisense polynucleotide of the present invention can be easily measured. As the radioisotope, for example, ³² P, ³ H and the like are used, and as the dye, for example, fluorescein, FAM (PE Bi
osystems), JOE (PE Biosystems), TA
MRA (made by PE Biosystems), ROX (PE Biosystems)
Fluorescent dyes such as Cy5 (manufactured by Amersham), Cy3 (manufactured by Amersham), and Cy3 (manufactured by Amersham). In addition, the amount of mRNA is
After conversion into NA, the DNA encoding apelin or apelin receptor or a part thereof or the antisense polynucleotide of the present invention is used as a primer P
This can be done by measuring the amount of cDNA amplified by CR. Thus, a test compound which increases the amount of mRNA encoding apelin or an apelin receptor can be selected as a substance having an activity of promoting the expression of apelin or an apelin receptor, and the apelin or apelin receptor can also be selected. A test compound that decreases the amount of mRNA encoding the protein can be selected as a substance having an activity of inhibiting the expression of apelin or an apelin receptor.

The present invention further provides (ii) the presence of a test compound in a transformant transformed with a recombinant DNA in which a reporter gene is ligated downstream of a promoter region or an enhancer region of a gene encoding apelin or apelin receptor. When cultured under and without
Provided is a method for screening a substance that promotes or inhibits the promoter activity, which comprises measuring and comparing the respective reporter activities. Examples of the reporter gene include lacZ (β-galactosidase gene), chloramphenicol acetyltransferase (CAT), luciferase, growth factor, β-glucuronidase, alkaline phosphatase, Green fluorescen.
t protein (GFP), β-lactamase and the like are used.
By measuring the amount of the reporter gene product (eg, mRNA, protein) using a known method, a test compound that increases the amount of the reporter gene product can control (particularly promote) the activity of the promoter or enhancer of the peptide of the present invention. Compound having the action of, i.e., a substance having an activity of promoting the expression of apelin or an apelin receptor. Conversely, a test compound that decreases the amount of the reporter gene product has a substance that has the effect of controlling (particularly inhibiting) the activity of the promoter or enhancer of apelin or the apelin receptor, that is, the activity of inhibiting the expression of apelin or the apelin receptor. It can be selected as a substance to have. The same test compounds as described above are used. Cultivation of the transformant can be performed in the same manner as in the above transformant.
The vector construction of the reporter gene and the assay method can follow known techniques (for example, Molecular Biotechn
ology 13, 29-43, 1999).

A substance having an activity of promoting the expression of apelin or an apelin receptor has an action of promoting the physiological activity of apelin, and is therefore useful as a safe and low-toxic pharmaceutical agent like apelin. On the contrary, a substance having an activity of inhibiting the expression of apelin or an apelin receptor can suppress the physiological activity of apelin, and is therefore useful as a safe and low-toxic pharmaceutical agent that suppresses the activity of apelin. Therefore, a substance that regulates the expression of apelin or an apelin receptor can be used as a drug such as an adrenocorticotropic hormone secretory regulator or an oxytocin secretory regulator. Specifically, a substance having an activity of promoting the expression of apelin or an apelin receptor, as an adrenocorticotropic hormone secretagogue, for example,
Hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease (fatigue, nausea,
Pigmentation, hypogonadism, hair loss, hypotension), adrenal insufficiency, pain, obesity, etc. can be used as a medicine for preventing or treating. Further, a substance having an activity of promoting the expression of apelin or an apelin receptor, as an oxytocin secretagogue, for example, a labor inducer, weak labor,
Used as a prophylactic / therapeutic agent for flaccid bleeding, uterine restoration failure, lactation failure, post-traumatic stress syndrome, milk stasis, etc. can do.

Substances having an activity of inhibiting the expression of apelin or an apelin receptor are, for example, Cushing's disease, Cushing's syndrome (central obesity, full-face like face, hypertension, purplish red skin line) as an adrenocorticotropic hormone secretion inhibitor. Streak, hirsutism), adrenocorticotropic hormone (ACTH) -producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism (primary and secondary), primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress,
It can be used as a medicine such as a prophylactic / therapeutic agent for depression, anorexia nervosa, gastric / duodenal ulcer, irritable bowel syndrome and the like. Further, a substance having an activity of inhibiting the expression of apelin or an apelin receptor is, as an oxytocin secretion inhibitor, for example, excessive labor pain, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth. , Prad
It can be used as a medicine such as a prophylactic / therapeutic agent for er-Willi syndrome and the like. The substance obtained by using the screening method of the present invention is selected from, for example, peptides, proteins, non-peptidic compounds, synthetic compounds, fermentation products, cell extracts, plant extracts, animal tissue extracts, plasma and the like. It is a compound. As the salt of the compound, the same salts as the above-mentioned salts of apelin are used. When the compound obtained by using the screening method of the present invention is used as the above-mentioned therapeutic / prophylactic agent, it can be carried out in a conventional manner. For example, tablets, capsules, elixirs, microcapsules, sterile solutions, in the same manner as the above-mentioned pharmaceutical composition containing apelin or a modified form thereof.
It can be a suspension agent or the like. Since the preparation thus obtained is safe and has low toxicity, for example, humans or warm-blooded animals (for example, mouse, rat, rabbit, sheep, pig, cow, horse, bird, cat, dog, monkey, chimpanzee, etc.) ). Although the dose of the compound or its salt varies depending on its action, target disease, administration subject, administration route, etc., for example, when a compound that promotes the expression of apelin is orally administered, it is generally an adult (body weight 60 kg Per day, the compound is about 0.1 to 100 mg, preferably about 1.0 to 50 mg, more preferably about 1.0 to 20 mg per day.
Administer. When administered parenterally, the single dose of the compound varies depending on the administration subject, target disease, etc.
For example, when a compound that promotes the expression of apelin is usually administered to an adult (per 60 kg) in the form of an injection, the compound is about 0.01 to 30 mg per day, preferably about 0.1 to 20 mg per day. More preferably about 0.1-1
It is convenient to administer about 0 mg intravenously. In the case of other animals, the dose converted per 60 kg can be administered.

(7) DNA Transfer Animal The present invention provides a DNA encoding an exogenous apelin or apelin receptor (hereinafter abbreviated as exogenous DNA of the present invention) or a mutant DNA thereof (exogenous mutated DNA of the present invention).
Sometimes abbreviated). That is, the present invention provides (i) the exogenous DN of the present invention.
A non-human mammal having A or a mutant DNA thereof, (i
i) the animal of (i) wherein the non-human mammal is a rodent, (iii) the animal of (ii) wherein the rodent is a mouse or rat, and (iv) the exogenous DNA of the present invention
Alternatively, the present invention provides a recombinant vector containing the mutant DNA and capable of being expressed in mammals. A non-human mammal having the exogenous DNA of the present invention or a mutant DNA thereof (hereinafter, abbreviated as the DNA-transferred animal of the present invention) is
For unfertilized eggs, fertilized eggs, germ cells including spermatozoa and their progenitor cells, preferably at the stage of embryogenesis in development of a non-human mammal (more preferably at the stage of single cells or fertilized egg cells and generally 8 Before the cell phase), the target DNA can be transferred by the calcium phosphate method, electric pulse method, lipofection method, agglutination method, microinjection method, particle gun method, DEAE-dextran method or the like. In addition, by the DNA transfer method, the target foreign DNA of the present invention can be transferred to somatic cells, living organs, tissue cells and the like and used for cell culture, tissue culture and the like. The DNA of the present invention is obtained by fusing the above-mentioned germ cells by a cell fusion method known per se.
It is also possible to create metastatic animals. Examples of the non-human mammal include cow, pig, sheep, goat, rabbit, dog, cat, guinea pig, hamster, mouse, rat and the like. Among them, rodents that have relatively short ontogenesis and biological cycle from the viewpoint of creating a pathological animal model system and are easy to reproduce, especially mice (for example, C57BL / 6 strain and DBA2 strain as pure strains). B6C3F ₁ system, BDF ₁ system, B
6D2F ₁ system, BALB / c system, ICR system, etc.)
Alternatively, a rat (eg, Wistar, SD, etc.) and the like are preferable. Examples of the “mammal” in the recombinant vector that can be expressed in mammals include humans in addition to the above-mentioned non-human mammals.

The exogenous DNA of the present invention is not the apelin or apelin receptor originally possessed by non-human mammals (hereinafter abbreviated as the DNA of the present invention), but once isolated and extracted from the mammal. And the DNA of the present invention. The mutant DNA of the present invention is a DNA having a mutation (eg, mutation) in the base sequence of the original DNA of the present invention, specifically, addition of a base, deletion, substitution with another base, etc. The generated DNA or the like is used, and abnormal DNA is also included. The abnormal DNA means a DNA that expresses an abnormal apelin or apelin receptor, and for example, a DNA that expresses a normal apelin or a peptide that suppresses the function of the apelin receptor is used. The exogenous DNA of the present invention may be derived from mammals of the same or different species as the target animal. When transferring the DNA of the present invention to a target animal, the DNA
It is generally advantageous to use as a DNA construct linked downstream of a promoter that can be expressed in animal cells. For example, when transferring the human DNA of the present invention,
The human of the present invention is placed downstream of various promoters capable of expressing DNA derived from various mammals (for example, rabbit, dog, cat, guinea pig, hamster, rat, mouse, etc.) having the DNA of the present invention having high homology. A DNA-transferred mammal highly expressing the DNA of the present invention can be produced by microinjecting a DNA construct (for example, a vector) having DNA bound thereto into a fertilized egg of a target mammal, for example, a mouse fertilized egg.

Expression vectors for apelin or apelin receptor include Escherichia coli-derived plasmids, Bacillus subtilis-derived plasmids, yeast-derived plasmids, bacteriophages such as λ phage, retroviruses such as Moloney leukemia virus, vaccinia virus or baculovirus. Animal viruses such as are used. Above all,
A plasmid derived from Escherichia coli, a plasmid derived from Bacillus subtilis, a plasmid derived from yeast, and the like are preferably used. Examples of the promoter for controlling the DNA expression include DNA-derived promoters derived from viruses (eg, simian virus, cytomegalovirus, Moloney leukemia virus, JC virus, breast cancer virus, poliovirus, etc.), various mammals (human , Rabbit, dog, cat, guinea pig, hamster, rat, mouse, etc.), eg, albumin, insulin I
I, uroplakin II, elastase, erythropoietin, endothelin, muscle creatine kinase, glial fibrillary acidic protein, glutathione S-transferase, platelet-derived growth factor β, keratins K1, K10 and K14, collagen type I and type II, cyclic AMP dependence Protein kinase βI subunit, dystrophin, tartrate-resistant alkaline phosphatase,
Atrial natriuretic factor, endothelial receptor tyrosine kinase (generally abbreviated as Tie2), sodium potassium adenosine 3 phosphatase (Na, K-ATPas)
e), neurofilament light chain, metallothionein I
And IIA, metalloproteinase 1 tissue inhibitor, MHC class I antigen (H-2L), H-ras, renin, dopamine β-hydroxylase, thyroid peroxidase (TPO), peptide chain elongation factor 1α (EF-1
α), β actin, α and β myosin heavy chains, myosin light chains 1 and 2, myelin basic protein, thyroglobulin, Thy-1, immunoglobulin, H chain variable region (VN
P), serum amyloid P component, myoglobin, troponin C, smooth muscle α-actin, preproenkephalin A, vasopressin and other promoters are used. Among them, cytomegalovirus promoter, human peptide chain elongation factor 1α (EF-1α) promoter, human and chicken β-actin promoter, which can be highly expressed systemically, are preferable. It is preferable that the vector has a sequence (generally called a terminator) that terminates the transcription of the desired messenger RNA in a DNA-transferred mammal,
For example, DNs derived from viruses and various mammals
The sequence A can be used, and preferably the SV40 terminator of simian virus is used.

In addition, for the purpose of further highly expressing the desired foreign DNA, the splicing signal of each DNA, the enhancer region, a part of the intron of eukaryotic DNA, and the like are provided 5 ′ upstream of the promoter region, between the promoter region and the translation region. Alternatively, it may be linked to the 3'downstream of the translation region depending on the purpose. A normal apelin or apelin receptor translation region includes liver-, kidney-, thyroid-cell, fibroblast-derived DNA derived from human or various mammals (eg, rabbit, dog, cat, guinea pig, hamster, rat, mouse) -derived DNA and Various commercially available genomic DNs
It can be obtained from the A library as the whole or a part of the genomic DNA, or the complementary DNA prepared from the liver, kidney, thyroid cell, and fibroblast-derived RNA by a known method as a raw material. In addition, the foreign abnormality D
NA can produce a translation region obtained by mutating a translation region of a normal peptide obtained from the above cells or tissues by point mutagenesis. The translation region can be prepared as a DNA construct that can be expressed in a transgenic animal by a conventional DNA engineering method in which it is linked to the downstream of the promoter and, if desired, the upstream of the transcription termination site. The transfer of the foreign DNA of the present invention at the fertilized egg cell stage is ensured to be present in all germinal cells and somatic cells of the target mammal. The presence of the exogenous DNA of the present invention in the embryonic cell of the produced animal after DNA transfer means that all the progeny of the produced animal retain the exogenous DNA of the present invention in all of its embryonic cells and somatic cells. means. The offspring of this type of animal that inherits the exogenous DNA of the present invention has the exogenous DNA of the present invention in all of its germ cells and somatic cells. The non-human mammal to which the foreign normal DNA of the present invention has been transferred has been confirmed to stably retain the foreign DNA by mating, and
A-carrying animals can be subcultured in a normal breeding environment. Exogenous DNA of the present invention at the fertilized egg cell stage
Metastases are ensured to be present in excess in all germinal and somatic cells of the subject mammal. Excessive presence of the exogenous DNA of the present invention in the germinal cells of the producing animal after DNA transfer means that all the progeny of the producing animal have the exogenous DNA of the present invention in all of their germinal cells and somatic cells. Means The offspring of this type of animal that inherited the exogenous DNA of the present invention have the exogenous DNA of the present invention in excess in all of their germ cells and somatic cells. Introduced DNA
By obtaining a homozygous animal having a homologous chromosome on both sides and crossing the male and female animals, all the progeny can be passaged so as to have the DNA in excess.

The non-human mammal having the normal DNA of the present invention is highly expressed with the normal DNA of the present invention, and by promoting the function of the endogenous normal DNA, finally the apelin or apelin receptor It may develop hyperfunction and can be used as a model animal for its pathological condition. For example, using the normal DNA-transferred animal of the present invention, elucidation of the pathological mechanism of hyperfunction of apelin or apelin receptor and diseases associated with apelin or apelin receptor and examination of treatment methods for these diseases are performed. It is possible. Further, since the mammal to which the exogenous normal DNA of the present invention has been transferred has a symptom of increased free apelin or apelin receptor, it can also be used in a screening test for a therapeutic drug for a disease associated with apelin or apelin receptor. It is possible. On the other hand, the non-human mammal having the exogenous abnormal DNA of the present invention can be subcultured as a DNA-carrying animal in a normal breeding environment after confirming that the exogenous DNA is stably retained by mating. Furthermore, the foreign DNA of interest can be incorporated into the above-mentioned plasmid and used as a raw material. The DNA construct with the promoter can be prepared by a usual DNA engineering technique. The transfer of the abnormal DNA of the present invention at the fertilized egg cell stage is ensured to be present in all germinal cells and somatic cells of the target mammal. The presence of the abnormal DNA of the present invention in the germinal cells of the producing animal after DNA transfer means that all the progeny of the producing animal have the abnormal DNA of the present invention in all of the germinal cells and somatic cells.
Is meant to have. The offspring of this type of animal that inherits the exogenous DNA of the present invention has the abnormal DNA of the present invention in all of its germ cells and somatic cells. Obtain a homozygous animal having the introduced DNA on both homologous chromosomes,
By mating the male and female animals, all offspring can be bred so that they have the DNA.

The non-human mammal having the abnormal DNA of the present invention has a high expression of the abnormal DNA of the present invention, and finally inhibits the function of the endogenous normal DNA, so that the aperin or the apelin receptor is finally expressed. It may become functionally inactive refractory and can be used as a model animal for the pathological condition. For example, by using the abnormal DNA-transferred animal of the present invention, it is possible to elucidate the pathological mechanism of the functionally inactive refractory disease of the peptide of the present invention and to examine the treatment method for this disease. In addition, as a specific utility, the animal with high expression of abnormal DNA of the present invention can be used as a normal apelin or normal apelin receptor due to abnormal apelin or abnormal apelin receptor in functionally inactive refractory aoriein or apelin receptor. It serves as a model to elucidate the inhibition of function (dominant negative action). In addition, since the mammal to which the foreign abnormal DNA of the present invention has been transferred has an increased symptom of the released apelin or apelin receptor of the present invention, a therapeutic drug screening for functionally inactive refractory apelin or apelin receptor is screened. It can also be used for testing. Further, other applications of the above-mentioned two kinds of the DNA-transferred animals of the present invention include, for example, use as a cell source for tissue culture, DNA or R in tissues of the DNA-transferred animals of the present invention.
Analysis of the relationship with peptides specifically expressed or activated by apelin or apelin receptor by directly analyzing NA or peptide tissues expressed by DNA, cells of tissues having DNA Are cultured by standard tissue culture techniques, and are used to study the function of cells from tissues that are generally difficult to culture, screening for agents that enhance the function of cells by using the cells described above, and the present invention It is possible to isolate and purify the mutant peptide of and to produce its antibody.

Further, the DNA-transferred animal of the present invention can be used to examine the clinical symptoms of apelin or apelin receptor-related diseases, including inactive insensitivity to apelin or apelin receptor, and the like. To obtain more detailed pathological findings in each organ of a disease model associated with apelin or apelin receptor, contribute to the development of new therapeutic methods, and further research and treatment of secondary diseases caused by the disease. You can Further, the DN of the present invention
It is possible to take out each organ from the A-transposed animal, cut it into small pieces, and then obtain the released DNA-transferred cells, culture the cells, or systematize the cultured cells with a proteolytic enzyme such as trypsin. Furthermore, it is possible to investigate the specification of apelin or apelin receptor-producing cells, their relation to apoptosis, differentiation or proliferation, or the signal transduction mechanism in them, and to investigate their abnormalities,
It becomes an effective research material for elucidation of apelin or apelin receptor and its action. Further, using the DNA-transferred animal of the present invention, for carrying out the development of a therapeutic agent for a disease associated with apelin or an apelin receptor, including a functionally inactive refractory apelin or an apelin receptor, the above-mentioned test method. It is possible to provide an effective and rapid screening method for the drug for treating the disease by using the quantification method and the like. In addition, it is possible to study and develop a DNA therapeutic method for diseases associated with apelin or apelin receptor using the DNA-transferred animal of the present invention or the foreign DNA expression vector of the present invention.

(8) Knockout Animal The present invention provides a non-human mammal embryonic stem cell in which the DNA of the present invention is inactivated and a DNA expression-deficient non-human mammal of the present invention. That is, the present invention provides (i) a non-human mammalian embryonic stem cell in which the DNA of the present invention is inactivated, (i
i) The DNA is a reporter gene (eg β derived from E. coli)
-The galactosidase gene) is inactivated by introducing the embryonic stem cell according to item (i), (iii) the embryonic stem cell according to item (i) that is resistant to neomycin, and (iv) the non-human mammal. The embryonic stem cell according to item (i) which is a rodent, (v) the embryonic stem cell according to item (iv) in which the rodent is a mouse, (vi) the DNA expression of the inactivated DNA of the present invention Deficient non-human mammal, (vii) The DNA is inactivated by introducing a reporter gene (eg, β-galactosidase gene derived from Escherichia coli), and the reporter gene is expressed under the control of a promoter for the DNA of the present invention. A non-human mammal according to paragraph (vi),
(Viii) The non-human mammal is a rodent (vi6)
The non-human mammal according to paragraph (ix), the rodent is a mouse, the non-human mammal according to paragraph (viii8), and (x) the animal according to paragraph (vii), the test compound being administered, Provided is a method for screening a compound or its salt that promotes or inhibits the promoter activity for DNA of the present invention, which comprises detecting the expression of a reporter gene. The non-human mammal embryonic stem cell in which the DNA of the present invention is inactivated means that the expression ability of the DNA is suppressed by artificially mutating the DNA of the present invention possessed by the non-human mammal. By substantially eliminating the activity of the peptide of the present invention encoded by the DNA, the DNA has substantially no ability to express the peptide of the present invention (hereinafter, this may be referred to as the knockout DNA of the present invention. ) Non-human mammal embryonic stem cells (hereinafter abbreviated as ES cells). As the non-human mammal, the same ones as described above are used. The method of artificially mutating the DNA of the present invention can be carried out, for example, by deleting a part or all of the DNA sequence or inserting or replacing another DNA by a genetic engineering technique. Due to these mutations, the knockout DNA of the present invention may be prepared, for example, by shifting the reading frame of the codon or disrupting the function of the promoter or exon.

Specific examples of the non-human mammalian embryonic stem cell in which the DNA of the present invention is inactivated (hereinafter abbreviated as the DNA inactivated ES cell of the present invention or the knockout ES cell of the present invention) include, for example, The DNA of the present invention contained in the target non-human mammal is isolated, and its exon portion has a neomycin resistance gene, a drug resistance gene represented by a hygromycin resistance gene, or lacZ (β-galactosidase gene), cat (chloram). A DNA sequence (eg, polyA) that disrupts the function of an exon by inserting a reporter gene typified by a phenicol acetyl transferase gene) or terminates gene transcription at an intron portion between exons.
Insert additional signals, etc., and complete messenger R
By disabling NA synthesis, a DNA chain having a DNA sequence constructed so as to destroy the gene as a result (hereinafter abbreviated as targeting vector) is introduced into the chromosome of the animal by, for example, homologous recombination method. , D on the Southern hybridization analysis or targeting vector of the obtained ES cells using the DNA sequence of the present invention or a DNA sequence in the vicinity thereof as a probe
The knockout E of the present invention was analyzed by the PCR method using the NA sequence and the DNA sequence of the neighboring region other than the DNA of the present invention used for the preparation of the targeting vector as a primer.
It can be obtained by selecting S cells. Also,
As the original ES cells for inactivating the DNA of the present invention by the homologous recombination method, for example, those already established as described above may be used, or newly established according to the method of known Evans and Kaufma. You can also use it. For example, in the case of mouse ES cells, currently, 129 ES cells are generally used.
Although cells have been used, the immunological background is not clear. For the purpose of obtaining an ES cell whose immunological genetic background is clear in a pure system as an alternative, for example, C
57BL / 6 mice and those established using BDF ₁ mice (F ₁ between C57BL / 6 and DBA / 2), in which the number of eggs collected by C57BL / 6 was improved by crossing with DBA / 2, are also good. Can be used. The BDF ₁ mouse has the advantage that the number of eggs collected is large and the eggs are strong, and since it has the C57BL / 6 mouse as a background, the ES cells obtained using this can be used as a pathological model mouse. , C57BL / 6 mice can be advantageously used in that its genetic background can be changed to that of C57BL / 6 mice. In addition, when ES cells are established, blastocysts on the 3.5th day after fertilization are generally used, but other than this, 8-cell stage embryos are ovulated and cultured up to the blastocysts for efficient and large numbers. The early embryos of can be obtained. Either male or female ES cells may be used, but male ES cells are usually more convenient for producing a germ line chimera. In addition, it is desirable to distinguish males and females as soon as possible in order to reduce troublesome culturing.

As a method for determining the sex of ES cells, for example, a method of amplifying and detecting a gene in the sex determining region on the Y chromosome by the PCR method can be mentioned. Using this method, conventionally, for example G-banding method, requires about 10 ⁶ cells for karyotype analysis, since suffices ES cell number of about 1 colony (about 50), culture It is possible to perform the primary selection of ES cells in the early stage by discriminating males and females, and the early selection of male cells can significantly reduce the labor required in the initial stage of culture.
The secondary selection can be performed, for example, by confirming the number of chromosomes by the G-banding method. The number of chromosomes of ES cells obtained is 100% of the normal number
However, if it is difficult because of physical manipulations during establishment, the gene of ES cell is knocked out, and then cloned again into normal cell (for example, cell having 2n = 40 chromosomes in mouse). Is desirable. The embryonic stem cell line thus obtained usually has a very good proliferative property, but since it is easy to lose the ability to develop ontogeny, it is necessary to carefully subculture. For example, on suitable feeder cells such as STO fibroblasts, LIF (1 to
Incubate in a carbon dioxide incubator (preferably 5% carbon dioxide, 95% air or 5% oxygen, 5% carbon dioxide, 90% air) in the presence of 10,000 U / ml at about 37 ° C. However, at the time of subculture, for example, trypsin / EDTA
Solution (usually 0.001-0.5% trypsin / 0.1-5)
mM EDTA, preferably about 0.1% trypsin / 1m
(M EDTA) treatment to obtain single cells and seeding on newly prepared feeder cells. Such subculture is usually performed every 1 to 3 days, but it is desirable to observe the cells at this time and to discard the cultured cells when morphologically abnormal cells are found. ES cells are differentiated into various types of cells such as parietal muscle, visceral muscle, and myocardium by performing monolayer culture until reaching a high density or suspension culture until forming a cell clump under appropriate conditions. Is possible [MJ Evans and MH
Kaufman, Nature Volume 292, 154, 1981.
Year; GR Martin Proceedings of National Academy of Sciences USA (Proc. Natl. Acad. Sci. USA) 78, 7634, 1
981; TC Doetschman et al., Journal of Embiology and Experimental Morphology, Vol. 87, p. 27, 1985], defective expression of DNA of the present invention obtained by differentiating ES cells of the present invention. The cells are useful in in vitro cell biology studies of the peptides of the invention or the receptor proteins of the invention.

The non-human mammal deficient in DNA expression of the present invention can be distinguished from a normal animal by measuring the mRNA level of the animal using a known method and indirectly comparing the expression level. is there. As the non-human mammal,
The same as the above is used. The non-human mammal deficient in DNA expression of the present invention can be obtained by, for example, introducing the targeting vector prepared as described above into mouse embryonic stem cells or mouse egg cells, and inactivating the DNA of the present invention in the targeting vector. The DNA of the present invention can be knocked out by homologous recombination in which the sequence replaces the DNA of the present invention on the chromosome of mouse embryonic stem cells or mouse egg cells by homologous recombination. A cell in which the DNA of the present invention has been knocked out is a DNA on or near the DNA of the present invention.
A DNA sequence on a Southern hybridization analysis or targeting vector using the sequence as a probe,
It can be determined by analysis by the PCR method using a DNA sequence of a region other than the mouse-derived DNA of the present invention used as the targeting vector as a primer. When a non-human mammal embryonic stem cell is used, a cell line in which the DNA of the present invention is inactivated by gene homologous recombination is cloned, and the cell is cloned at an appropriate time, for example, at the 8-cell stage. It is injected into an animal embryo or blastocyst, and the produced chimeric embryo is transplanted into the uterus of the pseudopregnant non-human mammal. The produced animal is a chimeric animal composed of both cells having the normal DNA locus of the present invention and cells having the artificially mutated DNA locus of the present invention. When a part of the germ cells of the chimeric animal has the mutated DNA locus of the present invention, all tissues are artificially mutated from the population obtained by mating such a chimeric individual with a normal individual. It can be obtained by selecting an individual composed of the cells having the added DNA locus of the present invention, for example, by judging the coat color. The individual thus obtained is usually an individual deficient in heteroexpression of the peptide of the present invention, and heterologous individuals deficient in heteroexpression of the peptide of the present invention are bred to homozygous expression of the peptide of the present invention from their offspring. Defective individuals can be obtained.

When egg cells are used, for example, a transgenic non-human mammal having a targeting vector introduced into its chromosome can be obtained by injecting a DNA solution into the nucleus of an egg cell by the microinjection method. It can be obtained by selecting those having a mutation in the DNA locus of the present invention by gene homologous recombination as compared with a transgenic non-human mammal. In this way, an individual whose DNA of the present invention has been knocked out can be passaged in a normal breeding environment after confirming that the DNA of the animal individual obtained by mating is also knocked out. Furthermore, the acquisition and maintenance of the germ line may be carried out according to the conventional method. That is, by mating male and female animals carrying the inactivated DNA,
A homozygous animal having the inactivated DNA on both homologous chromosomes can be obtained. The homozygous animals obtained are
It can be efficiently obtained by breeding a mother animal in a state where there are one normal individual and a plurality of homozygotes. By mating male and female heterozygous animals, homozygous and heterozygous animals having the inactivated DNA are bred. DNA of the present invention
The non-human mammal embryonic stem cells inactivated by the method are very useful for producing the non-human mammal deficient in DNA expression of the present invention. In addition, the non-human mammal deficient in DNA expression of the present invention lacks various biological activities that can be induced by the peptide of the present invention, and thus is a model of diseases caused by inactivation of biological activity of the peptide of the present invention. Therefore, it is useful for investigating the causes of these diseases and examining therapeutic methods.

(8a) Method for Screening Substance Having Curative / Preventive Effect Against Disease Caused by Defect or Damage of DNA of the Present Invention The non-human mammal deficient in DNA expression of the present invention is D
It can be used for screening a substance having a therapeutic / preventive effect on a disease caused by NA deficiency or damage. That is, the present invention results from a defect or damage of the DNA of the present invention, which comprises administering a test compound to a non-human mammal deficient in DNA expression of the present invention and observing and measuring the change in the animal. Provided is a method for screening a substance having a therapeutic / preventive effect on a disease. Examples of the non-human mammal deficient in DNA expression of the present invention used in the screening method include the same ones as described above. Examples of test compounds include peptides, proteins, non-peptidic compounds, synthetic compounds, fermentation products, cell extracts, plant extracts, animal tissue extracts, plasma, etc., and these compounds are novel compounds. Or a known compound may be used. Specifically, the non-human mammal deficient in DNA expression of the present invention is treated with a test compound and compared with a non-treated control animal, and tested using changes in each organ, tissue, disease symptoms, etc. of the animal as an index. The therapeutic / prophylactic effect of a compound can be tested. As a method of treating a test animal with a test compound, for example, oral administration, intravenous injection, etc. are used, and it can be appropriately selected depending on the symptoms of the test animal, the properties of the test compound, and the like. Also,
The dose of the test compound can be appropriately selected depending on the administration method, the properties of the test compound, and the like.

In the screening method, when the test compound is administered to the test animal, the function of the test animal is about 1
When the concentration is reduced by 0% or more, preferably about 30% or more, more preferably about 50% or more, the test compound can be selected as a substance having a therapeutic / prophylactic effect against the above-mentioned diseases. Specifically, the test compound has, for example, an adrenocorticotropic hormone secretion-regulating action, and is useful as an adrenocorticotropic hormone secretagogue, preferably an adrenocorticotropic hormone secretagogue, for example, low aldosterone. , Hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease (eg, fatigue, nausea, pigmentation, hypogonadism, hair loss, hypotension), adrenal insufficiency, pain, obesity, etc. It can be used as a medicine such as a therapeutic agent. Further, the test compound has an oxytocin secretion regulatory action, and as an oxytocin secretion regulator, preferably an oxytocin secretion promoter, for example, a labor inducer, weak labor, loose bleeding, uterine retrograde failure, lactation failure, trauma. It is useful as a preventive / therapeutic agent for post-stress syndrome, milk stasis, etc., or as an adjunct for pre- and post-placental delivery, cesarean section, miscarriage, artificial abortion, etc. In addition, the test compound can be applied as a test agent for investigating the secretory function of adrenocorticotropic hormone and the secretory function of oxytocin. The substance obtained using the screening method is a substance selected from the test compounds described above,
Since it has a therapeutic / prophylactic effect against a disease caused by apelin or apelin receptor deficiency or damage, it can be used as a medicine such as a safe and low-toxic therapeutic / prophylactic agent for the disease. Furthermore, compounds derived from the substances obtained by the above screening can be used as well.

The substance obtained by the screening method may form a salt, and the salt of the substance may be a physiologically acceptable acid (eg, inorganic acid, organic acid, etc.) or base (eg, salt). A salt with an alkali metal or the like) is used, and a physiologically acceptable acid addition salt is particularly preferable. Examples of such salts include salts with inorganic acids (for example, hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, etc.), or organic acids (for example, acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, Succinic acid, tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, etc.) and the like salts are used. The drug containing the substance obtained by the screening method can be produced in the same manner as the drug containing the apelin or a modified product thereof described above. Since the preparation thus obtained is safe and has low toxicity, it can be used, for example, in humans or mammals (for example, rat, mouse, guinea pig, rabbit, sheep, pig, cow, horse, cat, dog, monkey, etc.). Can be administered. The dose of the substance varies depending on the target disease, the subject to be administered, the administration route, etc. However, for example, when the substance is orally administered, an adult patient (body weight 60
(as kg), the compound is about 0.
1-100 mg, preferably about 1.0-50 mg, more preferably about 1.0-20 mg is administered. When administered parenterally, the single dose of the substance varies depending on the administration subject, target disease, etc. For example, when the substance is usually administered in the form of an injection to an adult patient (60 kg),
About 0.01 to 30 mg of the substance per day, preferably about 0.1 to 20 mg, more preferably about 0.1.
It is convenient to administer about 10 mg by intravenous injection. In the case of other animals, the dose converted per 60 kg can be administered.

(8b) Method for Screening Compound Promoting or Inhibiting Promoter Activity for DNA of the Present Invention The present invention provides a non-human mammal deficient in expression of the DNA of the present invention,
Provided is a method for screening a substance that promotes or inhibits the activity of a promoter for DNA of the present invention, which comprises administering a test compound and detecting the expression of a reporter gene. In the above screening method, the non-human mammal deficient in DNA expression of the present invention, among the non-human mammal deficient in DNA expression of the present invention, the DNA of the present invention is inactivated by introducing a reporter gene, A reporter gene that can be expressed under the control of a promoter for the DNA of the present invention is used. Examples of the test compound include the same ones as described above. As the reporter gene, the same ones as described above are used, and the β-galactosidase gene (lacZ), the soluble alkaline phosphatase gene, the luciferase gene and the like are preferable. In a non-human mammal deficient in expression of the DNA of the present invention in which the DNA of the present invention is replaced with a reporter gene, the reporter gene exists under the control of the promoter for the DNA of the present invention, and thus the expression of the substance encoded by the reporter gene is traced By doing so, the activity of the promoter can be detected. For example, a part of the DNA region encoding the peptide of the present invention is a β-galactosidase gene (lacZ) derived from Escherichia coli.
In the tissue where the peptide of the present invention is expressed, β-galactosidase is originally expressed in place of the peptide of the present invention. Thus, for example, 5-bromo-4-
Chloro-3-indolyl-β-galactopyranoside (X
It is possible to easily observe the expression state of the peptide of the present invention in the living body of an animal by staining with a reagent such as -gal) that serves as a substrate for β-galactosidase. Specifically, the peptide-deficient mouse of the present invention or a tissue section thereof is fixed with glutaraldehyde or the like, washed with phosphate buffered saline (PBS), and then stained with X-gal at room temperature or around 37 ° C. After reacting for about 30 minutes to 1 hour, the tissue sample was treated with 1 mM EDTA / P.
The β-galactosidase reaction may be stopped by washing with a BS solution, and the coloration may be observed. In addition, mRNA encoding lacZ may be detected according to a conventional method. The substance salt obtained by using the above-mentioned screening method is a substance selected from the above-mentioned test compounds, and is a substance that promotes or inhibits the promoter activity for the DNA of the present invention. The substance obtained by the screening method may form a salt, and the salt of the substance may be a physiologically acceptable acid (eg, inorganic acid) or base (eg, organic acid) or the like. Are used, and physiologically acceptable acid addition salts are particularly preferable. For such salt,
For example, inorganic acids (for example, hydrochloric acid, phosphoric acid, hydrobromic acid,
Sulfates) or organic acids (eg acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid,
Tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, etc.) and the like are used.

The substance that promotes the promoter activity for DNA of the present invention can promote the expression of apelin or apelin receptor and can promote the function of apelin, and therefore it is useful as, for example, an adrenocorticotropic hormone secretagogue. Yes, for example, hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease (eg, fatigue, nausea, pigmentation, hypogonadism, hair loss,
It can be used as a medicine such as a prophylactic / therapeutic agent for hypotension), adrenal insufficiency, pain, obesity and the like. Further, the substance that promotes the promoter activity for DNA of the present invention is, as an oxytocin secretion promoter, for example, a labor inducer, weak labor, relaxation bleeding, uterine restoration failure, lactation failure, posttraumatic stress syndrome, milk stasis, etc. It is useful as a prophylactic / therapeutic agent, or a medicine such as an auxiliary agent before and after delivery of placenta, cesarean section, miscarriage, artificial abortion and the like. On the other hand, the substance that inhibits the promoter activity for the DNA of the present invention can inhibit the expression of apelin or an apelin receptor and inhibit the function of apelin, and thus is useful as, for example, an adrenocorticotropic hormone secretion inhibitor. , For example, Cushing's disease, Cushing's syndrome (eg, central obesity, full-face like face, hypertension, violet skin striatum, hirsutism), adrenocorticotropic hormone (ACTH) -producing pituitary tumor, C
RH-producing tumor, aldosterone-producing tumor, aldosteronism (primary, secondary), primary cortisol resistance,
It can be used as a medicine such as a prophylactic / therapeutic agent for congenital adrenal enzyme deficiency, Addison's disease, stress, depression, anorexia nervosa, gastric / duodenal ulcer, irritable bowel syndrome and the like. Further, the substance that inhibits the promoter activity for DNA of the present invention is, as an oxytocin secretion inhibitor, for example, hypertonic labor, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth, Prader- It is useful as a medicine such as a preventive and therapeutic agent for Willi syndrome.
Further, substances derived from the substances obtained by the above screening can be used as well.

The drug containing the substance obtained by the screening method can be produced in the same manner as the drug containing the above-mentioned apelin or its modified form. Since the preparation thus obtained is safe and has low toxicity, for example, humans or mammals (for example, rat, mouse,
Guinea pig, rabbit, sheep, pig, cow, horse, cat, dog, monkey, etc.).
The dose of the substance varies depending on the target disease, subject to be administered, administration route, etc., but when a substance that promotes or inhibits the promoter activity for the DNA of the present invention is orally administered, it is generally an adult patient (body weight). (As 60 kg)
In the above, the amount of the substance is about 0.1 to 100 m per day.
g, preferably about 1.0-50 mg, more preferably about 1.0-20 mg. When administered parenterally, the single dose of the substance varies depending on the administration subject, target disease, etc., but for example, a substance that promotes or inhibits the promoter activity for the DNA of the present invention is usually in the form of an injection. When administered to an adult patient (as 60 kg), the substance is about 0.01 to 30 mg per day, preferably about 0.1 to 20 mg, more preferably about 0.1 to 1 mg.
It is convenient to administer about 0 mg intravenously. In the case of other animals, the dose converted per 60 kg can be administered.

As described above, the non-human mammal deficient in expression of the DNA of the present invention is extremely useful for screening a compound or its salt that promotes or inhibits the activity of the promoter for the DNA of the present invention, and the DNA of the present invention. It can greatly contribute to the investigation of the causes of various diseases caused by the expression failure or the development of preventive / therapeutic drugs. In addition, by using DNA containing the promoter region of the peptide of the present invention, genes encoding various proteins are ligated downstream thereof, and this is injected into an egg cell of an animal to obtain a so-called transgenic animal (transgenic animal). Once created, it is possible to specifically synthesize the peptide and study its action in the living body. Further, by binding an appropriate reporter gene to the promoter portion and establishing a cell line in which it is expressed, a low-molecular compound having the action of specifically promoting or suppressing the in vivo production ability of the peptide itself of the present invention. Can be used as a search system for.

In the present specification and drawings, when an abbreviation is used for a base, amino acid, etc., IUPAC-IUB is used.
Based on the abbreviations used in Commision on Biochemical Nomenclature or conventional abbreviations in this field,
An example is given below. When amino acids may have optical isomers, the L form is shown unless otherwise specified.

[0124]   DNA: Deoxyribonucleic acid   cDNA: Complementary deoxyribonucleic acid     A: Adenine     T: thymine     G: Guanine     C: cytosine     Y: thymine or cytosine     N: thymine, cytosine, adenine or guanine     R: adenine or guanine     M: cytosine or adenine     W: Thymine or adenine     S: Cytosine or guanine   RNA: Ribonucleic acid   mRNA: messenger ribonucleic acid   dATP: Deoxyadenosine triphosphate   dTTP: Deoxythymidine triphosphate   dGTP: Deoxyguanosine triphosphate   dCTP: Deoxycytidine triphosphate   ATP: Adenosine triphosphate   EDTA: ethylenediaminetetraacetic acid   SDS: Sodium dodecyl sulfate   TFA: trifluoroacetic acid   EIA: Enzyme immunoassay

[0125]   Gly or G: Glycine   Ala or A: Alanine   Val or V: Valine   Leu or L: Leucine   Ile or I: Isoleucine   Ser or S: Serine   Thr or T: threonine   Cys or C: cysteine   Met or M: methionine   Glu or E: Glutamic acid   Asp or D: Aspartic acid   Lys or K: lysine   Arg or R: Arginine   His or H: Histidine   Phe or F: Phenylalanine   Tyr or Y: Tyrosine   Trp or W: Tryptophan   Pro or P: Proline   Asn or N: Asparagine   Gln or Q: Glutamine   pGlu: pyroglutamic acid   Me: Methyl group   Et: ethyl group   Bu: butyl group   Ph: Phenyl group   TC: thiazolidine-4 (R) -carboxamide group   Bom: benzyloxymethyl   NMP: N-methylpyrrolidone   PAM: Phenylacetamidomethyl

Further, substituents, protecting groups and reagents commonly used in the present specification are represented by the following symbols. Tos: p-toluenesulfonyl HONB: N-hydroxy-5-norbornene-2,3
-Dicarboximide Bzl: benzyl Z: benzyloxycarbonyl Br-Z: 2-bromobenzyloxycarbonyl Cl-Z: 2-chlorobenzyloxycarbonyl Boc: t-butyloxycarbonyl HOBt: 1-hydroxybenztriazole DCC: N, N'-dicyclohexylcarbodiimide TFA: trifluoroacetic acid Fmoc: N-9-fluorenylmethoxycarbonyl DNP: dinitrophenyl Bum: tertiary butoxymethyl Trt: trityl MeBzl: 4-methylbenzyl

The sequence numbers in the sequence listing in the present specification show the following sequences. [SEQ ID NO: 1] This shows the amino acid sequence of bovine apelin (17 sequences from the N terminus). [SEQ ID NO: 2] This shows the base sequence of cDNA encoding bovine apelin having the amino acid sequence represented by SEQ ID NO: 1. [SEQ ID NO: 3] This shows the entire amino acid sequence of the apelin receptor encoded by the apelin receptor cDNA. [SEQ ID NO: 4] This shows the entire base sequence of apelin receptor cDNA. [SEQ ID NO: 5] cDNA encoding mouse apelin
The amino acid sequence encoded by A is shown. [SEQ ID NO: 6] cDNA encoding mouse apelin
The base sequence of A is shown. [SEQ ID NO: 7] cDNA encoding rat apelin
The amino acid sequence encoded by A is shown. [SEQ ID NO: 8] cDNA encoding rat apelin
The base sequence of A is shown. [SEQ ID NO: 9] cDNA encoding human apelin
The amino acid sequence encoded by [SEQ ID NO: 10] cDNA encoding human apelin
The base sequence of A is shown. [SEQ ID NO: 11] cDNA encoding bovine apelin
The amino acid sequence encoded by A is shown. [SEQ ID NO: 12] cDNA encoding bovine apelin
The base sequence of A is shown. [SEQ ID NO: 13] to [SEQ ID NO: 52] This shows the amino acid sequence of a specific peptide of the modified apelin (I). [SEQ ID NO: 53] to [SEQ ID NO: 119] This shows the amino acid sequences of the specific peptides of the modified apelin (II) to (IV).

[0128]

The present invention will be described in more detail with reference to the following examples, which are not intended to limit the scope of the present invention.

Example 1 Effect on the Pituitary Hormone Concentration of Apelin The effect of the administration of apelin on the pituitary hormone level in plasma was examined. Adult Wistar male rats (body weight during surgery: 320 to 350 g) were anesthetized by intraperitoneal administration of pentobarbital 50 mg / kg, and fixed to a rat stereotaxic apparatus. The incisor bar was 3.3 mm lower than the interoral line. The skull is exposed and the guide cannula AG-8 (inner diameter 0.4 mm, outer diameter 0.5) is placed in the third ventricle.
mm, ecom) was drilled into the bone using a dental drill. In addition, anchor screws were buried in three places around it. Stainless steel guide cannula, AG-8
Was inserted so that its tip was located above the third ventricle. The stereotaxic coordinates are AP: -0.8mm, L: 0.0 according to Bregma according to Atlas of Paxinos and Watson (1998).
mm, H: -5.4 mm. The guide cannula was fixed to the skull with dental cement and anchor screws.
The guide cannula has a stainless steel dummy cannula,
AD-8 (outer diameter 0.35 mm, Acom) was inserted and fixed with a cap nut (Acom). After surgery, the rats were housed in individual cages. After implanting the guide cannula for about 1 week and waiting for post-operative recovery, remove the cap nut and dummy cannula attached to the rat's skull, and replace it with a Teflon (registered trademark) tube (length 50 cm, inner diameter 0.1 mm). , Outer diameter 0.35mm, stainless steel micro-injection cannula AMI-9 (inner diameter 0.17mm, outer diameter 0.35)
mm, E.com). The length of the microinjection cannula was adjusted so that the tip 1 mm was exposed from the guide cannula. One of the Teflon (registered trademark) tubes was connected to a microsyringe pump, and phosphate buffered saline (pH 7.2) or rat apelin (the amino acid sequence from the 42nd to the 77th amino acid sequence represented by SEQ ID NO: 7) was used. Peptide) (1
5 μL of phosphate buffered saline in which 0 nmol) was dissolved
A total of 10 μL was injected into the lateral ventricle at a minute flow rate. After waiting 5 minutes after completion of the injection, the microinjection cannula was removed, and the dummy cannula was fixed again with a cap nut. Ten minutes after the completion of the injection, the animals were decapitated and blood was collected. Blood is 300 KI containing 3 mg / ml EDTA in a 50 ml tube (Corning) to prevent coagulation.
300 μL of U / ml aprotinin solution was placed. The collected blood was centrifuged (2,500 rpm, 25 minutes) using a high-speed cooling centrifuge (HIMAC 5DL, Hitachi, Ltd.) to collect a supernatant (plasma). Plasma adrenocorticotropic hormone (ACTH), luteinizing hormone (LH), thyroid stimulating hormone (TSH), follicle stimulating hormone (FSH), prolactin, oxytocin and arginine-vasopressin (Ar)
g-Vasopressin)) is a commercially available radioimmunoassay (ACTH
And Arg-Vasopressin; Yuka Medias, LH, TS
H, FSH and prolactin; Amersham). In addition, oxytocin can be prepared by a previously reported method [Neuroscience Letters, Vol. 285, p. 234 (2000
Year)]. As shown in FIG. 1, the apelin administration group showed a significant increase only in blood ACTH and oxytocin concentrations as compared with the control group.

Example 2 Effect of Apelin on ACTH Concentration With respect to the effect of intracerebroventricular administration of apelin on ACTH concentration in plasma, changes with time after administration of apelin were examined. Adult Wistar male rats treated in the same manner as in Example 1 were implanted with a guide cannula, and were bred for about 1 week, awaited recovery after the operation, and operated for free blood collection. Rats that underwent the above operation were pentobarbital 5
Anesthesia was performed by intraperitoneal administration of 0 mg / kg. The dorsal position was fixed on the dissection pad, and the left jugular vein was exposed. Polyethylene tube SP35 (inner diameter 0.5 mm, outer diameter 0.
9mm, Natsume Seisakusho), cut into about 30cm length, 20
After filling with 0 unit / ml of heparin-containing physiological saline, it was inserted into the jugular vein for about 4.5 cm and fixed. The other end of the tube was subcutaneously exposed on the dorsal side and exposed from the neck (dorsal side).
After waiting overnight after the operation, 300 μL of blood was collected before administration of apelin using a tuberculin syringe with a dose of 1 ml and a 25 gauge needle (both are Terumo Corp.). In order to prevent blood coagulation, 10 μL of 300 KIU / ml aprotinin solution containing 3 mg · ml EDTA was previously put in the syringe.
Remove the capnite and dummy cannula attached to the rat skull, and replace it with a Teflon (registered trademark) tube (length 50 cm, inner diameter 0.1 mm, outer diameter 0.35 m).
m, Eicom) stainless steel microinjection cannula (inner diameter 0.17 mm, outer diameter 0.
35 mm, E.com) was inserted. The length of the microinjection cannula was adjusted so that the tip 1 mm was exposed from the guide cannula. One side of a Teflon (registered trademark) tube was connected to a microsyringe pump, and phosphate buffered saline (pH 7.2) or rat apelin (amino acids 42 to 77 in the amino acid sequence represented by SEQ ID NO: 7) Sequence-containing peptide) (3,10 nmol) at a flow rate of 5 μL / min for a total of 1
0 μL was injected into the third ventricle. After waiting 5 minutes after the completion of the injection, the microinjection cannula was removed, and the dummy cannula was fixed again with the capnite. 300 μL of blood was collected from the jugular vein 0, 10, 20, 30, 60 minutes after the start of intraventricular administration. The collected blood was centrifuged (13,000 rpm, 5 minutes) using a micro high speed cooling centrifuge (MR-150, Tommy Seiko), and the supernatant (plasma) was collected. ACTH contained in plasma was measured using a radioimmunoassay (Yuka Medias). As shown in FIG. 2, the apelin-administered group was higher than the apelin-n group in comparison with the control group.
A rising tendency was observed in the mol administration group, and apelin 10 nm
In the ol-administered group, a significant increase in blood ACTH concentration was observed at any blood collection point, with a peak at 10 minutes after administration.

Example 3 Effect of Apelin on Oxytocin Concentration The present inventors examined changes over time after administration of apelin in the effect of third intracerebroventricular administration of apelin on oxytocin concentration in plasma. Rat apelin (42nd amino acid sequence represented by SEQ ID NO: 7) as in Example 2
Peptide having the amino acid sequence from the 77th position to the 77th position)
After the administration, blood was collected and plasma was collected. ACTH contained in plasma was measured using a radioimmunoassay (Yuka Medias). As shown in FIG. 3, the apelin administration group showed a significant increase in blood oxytocin concentration 10 to 20 minutes after administration in the apelin 10 nmol administration group as compared with the control group.

[0132]

EFFECTS OF THE INVENTION Apelin or its modified form has an adrenocorticotropic hormone secretory regulating action and an oxytocin secretory regulating action, and is useful as an adrenocorticotropic hormone secretory regulating agent and an oxytocin secretory regulating agent. Also,
Apelin or apelin receptor is useful for screening adrenocorticotropic hormone or oxytocin secretion regulators.

[0133]

[SEQUENCE LISTING]

<110> Takeda Chemical Industries, Ltd. <120> Use of Apelin <130> B03013 <150> JP 2002-020031 <151> 2002-01-29 <160> 119 <210> 1 <211> 17 <212> PRT <213> Bovine <400> 1 Leu Val Gln Pro Arg Gly Pro Arg Ser Gly Pro Gly Pro Trp Gln Gly 1 5 10 15 Gly 17 <210> 2 <211> 51 <212> DNA <213> Bovine <400> 2 ytngtncarc cnmgnggncc nmgnwsnggn ccnggnccnt ggcarggngg n 51 <210> 3 <211> 380 <212> PRT <213><400> 3 Met Glu Glu Gly Gly Asp Phe Asp Asn Tyr Tyr Gly Ala Asp Asn Gln 1 5 10 15 Ser Glu Cys Glu Tyr Thr Asp Trp Lys Ser Ser Gly Ala Leu Ile Pro 20 25 30 Ala Ile Tyr Met Leu Val Phe Leu Leu Gly Thr Thr Gly Asn Gly Leu 35 40 45 Val Leu Trp Thr Val Phe Arg Ser Ser Arg Glu Lys Arg Arg Ser Ala 50 55 60 Asp Ile Phe Ile Ala Ser Leu Ala Val Ala Asp Leu Thr Phe Val Val 65 70 75 80 Thr Leu Pro Leu Trp Ala Thr Tyr Thr Tyr Arg Asp Tyr Asp Trp Pro 85 90 95 Phe Gly Thr Phe Phe Cys Lys Leu Ser Ser Tyr Leu Ile Phe Val Asn 100 105 110 Met Tyr Ala Ser Val Phe Cys Leu Thr Gly Leu Ser Phe Asp Arg Tyr 115 120 125 Leu Ala Ile Val Arg Pro Val Ala Asn Ala Arg Leu Arg Leu Arg Val 130 135 140 Ser Gly Ala Val Ala Thr Ala Val Leu Trp Val Leu Ala Ala Leu Leu 145 150 155 160 Ala Met Pro Val Met Val Leu Arg Thr Thr Gly Asp Leu Glu Asn Thr 165 170 175 Thr Lys Val Gln Cys Tyr Met Asp Tyr Ser Met Val Ala Thr Val Ser 180 185 190 Ser Glu Trp Ala Trp Glu Val Gly Leu Gly Val Ser Ser Thr Thr Val 195 200 205 Gly Phe Val Val Pro Phe Thr Ile Met Leu Thr Cys Tyr Phe Phe Ile 210 215 220 Ala Gln Thr Ile Ala Gly His Phe Arg Lys Glu Arg Ile Glu Gly Leu 225 230 235 240 Arg Lys Arg Arg Arg Leu Leu Ser Ile Ile Val Val Leu Val Val Thr 245 250 255 Phe Ala Leu Cys Trp Met Pro Tyr His Leu Val Lys Thr Leu Tyr Met 260 265 270 Leu Gly Ser Leu Leu His Trp Pro Cys Asp Phe Asp Leu Phe Leu Met 275 280 285 Asn Ile Phe Pro Tyr Cys Thr Cys Ile Ser Tyr Val Asn Ser Cys Leu 290 295 300 Asn Pro Phe Leu Tyr Ala Phe Phe Asp Pro Arg Phe Arg Gln Ala Cys 305 310 315 320 Thr Ser Met Leu Cys Cys Gly Gln Ser Arg Cys Ala Gly Thr Ser His 325 330 335 Ser Ser Ser Gly Glu Lys Ser Ala Ser Tyr Ser Ser Gly His Ser Gln 340 345 350 Gly Pro Gly Pro Asn Met Gly Lys Gly Gly Glu Gln Met His Glu Lys 355 360 365 Ser Ile Pro Tyr Ser Gln Glu Thr Leu Val Val Asp 370 375 380 <210> 4 <211> 1140 <212> DNA <213> Bovine <400> 4 atggaggaag gtggtgattt tgacaactac tatggggcag acaaccagtc tgagtgtgag 60 tacacagact ggaaatcctc gggggccctc atccctgcca tctacatgtt ggtcttcctc 120 ctgggcacca cgggaaacgg tctggtgctc tggaccgtgt ttcggagcag ccgggagaag 180 aggcgctcag ctgatatctt cattgctagc ctggcggtgg ctgacctgac cttcgtggtg 240 acgctgcccc tgtgggctac ctacacgtac cgggactatg actggccctt tgggaccttc 300 ttctgcaagc tcagcagcta cctcatcttc gtcaacatgt acgccagcgt cttctgcctc 360 accggcctca gcttcgaccg ctacctggcc atcgtgaggc cagtggccaa tgctcggctg 420 aggctgcggg tcagcggggc cgtggccacg gcagttcttt gggtgctggc cgccctcctg 480 gccatgcctg tcatggtgtt acgcaccacc ggggacttgg agaacaccac taaggtgcag 540 tgctacatgg actactccat ggtggccact gtgagctcag agtgggcctg ggaggtgggc 600 cttggggtct cgtccaccac cgtgggcttt gtggtgccct tc accatcat gctgacctgt 660 tacttcttca tcgcccaaac catcgctggc cacttccgca aggaacgcat cgagggcctg 720 cggaagcggc gccggctgct cagcatcatc gtggtgctgg tggtgacctt tgccctgtgc 780 tggatgccct accacctggt gaagacgctg tacatgctgg gcagcctgct gcactggccc 840 tgtgactttg acctcttcct catgaacatc ttcccctact gcacctgcat cagctacgtc 900 aacagctgcc tcaacccctt cctctatgcc tttttcgacc cccgcttccg ccaggcctgc 960 acctccatgc tctgctgtgg ccagagcagg tgcgcaggca cctcccacag cagcagtggg 1020 gagaagtcag ccagctactc ttcggggcac agccaggggc ccggccccaa catgggcaag 1080 ggtggagaac agatgcacga gaaatccatc ccctacagcc aggagaccct tgtggttgac 1140 <210> 5 <211> 77 <212> PRT <213> Mouse <400> 5 Met Asn Leu Arg Leu Cys Val Gln Ala Leu Leu Leu Leu Trp Leu Ser 1 Thr Ala Val Cys Gly Val Pro Leu Met Leu Pro Pro Asp Gly Thr 20 25 30 Gly Leu Glu Glu Gly Ser Met Arg Tyr Leu Val Lys Pro Arg Thr Ser 35 40 45 Arg Thr Gly Pro Gly Ala Trp Gln Gly Gly Arg Arg Lys Phe Arg Arg 50 55 60 Gln Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 65 70 75 <210> 6 <211 > 231 <212> DNA <213> Mouse <400> 6 atgaatctga ggctctgcgt gcaggcgctg ctgctgctct ggctctcctt gactgcagtt 60 tgtggagtgc cactgatgtt gcctccagat ggaacaggac tagaagaagg aagcatgcgc 120 tacctggtga agcccagaac ttcgaggact ggaccaggag cctggcaggg aggcaggagg 180 aaatttcgca gacagcgccc ccggctctcc cataagggcc ccatgccttt c 231 <210> 7 <211> 77 <212> PRT <213> Rat <400> 7 Met Asn Leu Ser Phe Cys Val Gln Ala Leu Leu Leu Leu Trp Leu Ser 1 5 10 15 Leu Thr Ala Val Cys Gly Val Pro Leu Met Leu Pro Pro Asp Gly Lys 20 25 30 Gly Leu Glu Glu Gly Asn Met Arg Tyr Leu Val Lys Pro Arg Thr Ser 35 40 45 Arg Thr Gly Pro Gly Ala Trp Gln Gly Gly Arg Arg Lys Phe Arg Arg 50 55 60 Gln Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 65 70 75 <210> 8 <211> 231 <212> DNA <213> Rat <400> 8 atgaatctga gtttctgcgt gcaggcgctg ctgctgctct ggctctcctt gactgccgtg 60 tgtggagtgc cactgatgct gcctccagat gggaaagggc tagaagaagg caacatgcgc 120 tacctggtga agcccagaac ttcgaggact ggaccagggg cctggcaggg aggcaggagg 180 aaatttcgca gacagcggcc ccgtctctcc cataagg gac ccatgccttt c 231 <210> 9 <211> 77 <212> PRT <213> Human <400> 9 Met Asn Leu Arg Leu Cys Val Gln Ala Leu Leu Leu Leu Trp Leu Ser 1 5 10 15 Leu Thr Ala Val Cys Gly Gly Ser Leu Met Pro Leu Pro Asp Gly Asn 20 25 30 Gly Leu Glu Asp Gly Asn Val Arg His Leu Val Gln Pro Arg Gly Ser 35 40 45 Arg Asn Gly Pro Gly Pro Trp Gln Gly Gly Arg Arg Lys Phe Arg Arg 50 55 60 Gln Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 65 70 75 <210> 10 <211> 231 <212> DNA <213> Human <400> 10 atgaatctgc ggctctgcgt gcaggcgctc ctgctgctct ggctctcctt gccgcggtg 60 tgtggggatgatgatg 120 cacctggtgc agcccagagg gtcaaggaat gggccagggc cctggcaggg aggtcggagg 180 aaattccgcc gccagcggcc ccgcctctcc cataagggac ccatgccttt c Le 231 <210> 11 <211> 77 <212> PRT <213> Bovine Asu Leu Arn Leuet 4001 Cys 1 5 10 15 Leu Ser Ala Val Cys Gly Gly Pro Leu Leu Gln Thr Ser Asp Gly Lys 20 25 30 Glu Met Glu Glu Gly Thr Ile Arg Tyr Leu Val Gln Pro Arg Gly P ro 35 40 45 Arg Ser Gly Pro Gly Pro Trp Gln Gly Gly Arg Arg Lys Phe Arg Arg 50 55 60 Gln Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 65 70 75 <210> 12 <211> 231 <212> DNA <213> Bovine <400> 12 atgaatctgc ggcgctgcgt gcaggcgctc ctgctgctct ggctctgcct gagcgcggtg 60 tgcggaggac ccctgctgca gacttctgac gggaaggaga tggaagaagg caccatccga 120 tacctggtgc agcccagggg gccgaggagc ggcccaggcc cctggcaggg aggtcggagg 180 aagttccggc gccagcggcc acgcctctcc cacaagggtc ccatgccttt c 231 <210> 13 <211> 36 <212> PRT <213> Artificial Sequence <223> Xaa means Nle. <400> 13 Leu Val Gln Pro Arg Gly Ser Arg Asn Gly Pro Gly Pro Trp Gln Gly 1 5 10 15 Gly Arg Arg Lys Phe Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly 20 25 30 Pro Xaa Pro Phe 35 <210> 14 <211> 36 <212> PRT <213> Artificial Sequence <223> Xaa means Nle. <400> 14 Leu Val Gln Pro Arg Gly Ser Arg Asn Gly Pro Gly Pro Trp Gln Gly 1 5 10 15 Gly Arg Arg Lys Phe Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly 20 25 30 Pro Xaa Pro Tyr 35 <210> 15 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Nle. <400> 15 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Phe 1 5 10 <210> 16 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Nle. <400> 16 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Tyr 1 5 10 <210> 17 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Nle. <400> 17 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro 1 5 10 <210> 18 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Nle. <400> 18 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa 1 5 10 <210> 19 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means Ac-Arg, Xaa on the 10th position mea ns Nle. <400> 19 Xaa Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Tyr 1 5 10 <210> 20 <211> 11 <212> PRT <213> Artificial S sequence <223> Xaa on the 1st position means Ac-Arg, Xaa on the 10th position mea ns Nle. <400> 20 Xaa Pro Arg Leu Ser His Lys Gly Pro Xaa Pro 1 5 10 <210> 21 <211> 10 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means Ac-Arg, Xaa on the 10th position mea ns Nle. <400> 21 Xaa Pro Arg Leu Ser His Lys Gly Pro Xaa 1 5 10 <210> 22 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 8th position means Lys (Ac). <400> 22 Xaa Arg Pro Arg Leu Ser His Xaa Gly Pro Met Pro Phe 1 5 10 <210> 23 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 8th position means Lys (Me). <400> 23 Xaa Arg Pro Arg Leu Ser His Xaa Gly Pro Met Pro Phe 1 5 10 <210> 24 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 8th position means Lys (Ac), Xaa on the 11th position means Nle. <400> 24 Xaa Arg Pro Arg Leu Ser His Xaa Gly Pro Xaa Pro Phe 1 5 10 <210> 25 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 8th position means Lys (Me), Xaa on the 11th position means Nle. <400> 25 Xaa Arg Pro Arg Leu Ser His Xaa Gly Pro Xaa Pro Phe 1 5 10 <210> 26 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 8th position means Lys (Tos), Xaa on the 11th position means Nle. <400> 26 Xaa Arg Pro Arg Leu Ser His Xaa Gly Pro Xaa Pro Phe 1 5 10 <210> 27 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 8th position means Arg (Tos), Xaa on the 11th position means Nle. <400> 27 Xaa Arg Pro Arg Leu Ser His Xaa Gly Pro Xaa Pro Phe 1 5 10 <210> 28 <211 > 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 5th position means Nle, Xaa on the 11th position means Nle. <400> 28 Xaa Arg Pro Arg Xaa Ser His Lys Gly Pro Xaa Pro Phe 1 5 10 <210> 29 <211> 13 <212> PRT <213> Artificial Sequence <223> X aa on the 1st position means pGlu, Xaa on the 5th position means Nle, Xaa on the 11th position means Nle. <400> 29 Xaa Arg Pro Arg Xaa Ser His Lys Gly Pro Xaa Pro Tyr 1 5 10 <210> 30 <211 > 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Nle, Xaa on the 13th position means Thi. <400> 30 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Xaa 1 5 10 <210> 31 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Nle, Xaa on the 13th position means Phg. <400> 31 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Xaa 1 5 10 <210> 32 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Nle, Xaa on the 13th position means Pya (2). <400> 32 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Xaa 1 5 10 <210> 33 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Nle. <400> 33 Arg Pro Ar g Leu Ser His Lys Gly Pro Nle Pro Tyr 1 5 10 <210> 34 <211> 36 <212> PRT <213> Artificial Sequence <223> Xaa on the 3rd position means Adi (NH ₂ ), Xaa on the 34th position m eans Nle. <400> 34 Leu Val Xaa Pro Arg Gly Ser Arg Asn Gly Pro Gly Pro Trp Gln Gly 1 5 10 15 Gly Arg Arg Lys Phe Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly 20 25 30 Pro Xaa Pro Phe 35 <210> 35 <211> 36 <212> PRT <213> Artificial Sequence <223> Xaa on the 3rd position means Lys (Ac), Xaa on the 34th position me ans Nle. <400> 35 Leu Val Xaa Pro Arg Thr Ser Arg Thr Gly Pro Gly Ala Trp Gln Gly 1 5 10 15 Gly Arg Arg Lys Phe Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly 20 25 30 Pro Xaa Pro Tyr 35 <210> 36 <211> 37 <212 > PRT <213> Artificial Sequence <223> Xaa on 35th position means Nle. <400> 36 Tyr Leu Val Lys Pro Arg Thr Ser Arg Thr Gly Pro Gly Ala Trp Gln 1 5 10 15 Gly Gly Arg Arg Lys Phe Arg Arg Gln Arg Pro Arg Leu Ser His Lys 20 25 30 Gly Pro Xaa Pro Phe 35 <210> 37 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means Z-pGlu, Xaa on the 8th position mean s Lys (Ac), Xaa on the 11th position means Nle. <400> 37 Xaa Arg Pro Arg Leu Ser His Xaa Gly Pro Xaa Pro Phe 1 5 10 <210> 38 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 13th position means Met (O). <400> 38 Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly Pro Xaa 1 5 10 <210> 39 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa on the 13th position means Nle. <400> 39 Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Tyr 1 5 10 15 <210> 40 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 12th position means Phe (Cl). <400> 40 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 41 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 13th position means Phe (Cl). <400> 41 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Xaa 1 5 10 <210> 42 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 12th position means Nal (2). <400> 42 Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Xaa 1 5 10 <210> 43 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa on the 11th position means Nal (2). <400> 43 Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 44 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 13th position means Phe (Cl). <400> 44 Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Xaa 1 5 10 <210> 45 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa on the 11th position means Phe (Cl). <400> 45 Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 46 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa on the 11th position means Cha. <400> 46 Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 47 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Cha. <400> 47 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Phe 1 5 10 <210> 48 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on th e 13th position means Cha. <400> 48 Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly Pro Xaa 1 5 10 <210> 49 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa on the 15th position means Phe (Cl). <400> 49 Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Xaa 1 5 10 15 <210> 50 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Nle, Xaa on the 15th position means Phe (Cl). <400> 50 Arg Arg Gln Arg Pro Arg Leu Ser His Xaa Gly Pro Met Pro Xaa 1 5 10 15 <210> 51 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Nle, Xaa on the 15th position means Tyr (I). <400> 51 Arg Arg Gln Arg Pro Arg Leu Ser His Xaa Gly Pro Met Pro Xaa 1 5 10 15 <210> 52 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Nle, Xaa on the 15th position means Tyr (Me). <400> 52 Arg Arg Gln Arg Pro Arg Leu Ser His Nle Gly Pro Met Pro Xaa 1 5 10 15 <210> 53 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa means Phe (Cl) <400 > 53 Arg Arg Gln Arg Pro Arg Leu Ser Ala Arg Gly Pro Met Pro Xaa 1 5 10 15 <210> 54 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa on the 13th position means Cha, Xaa on the 15th position means Phe (Cl) <400> 54 Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly Pro Cha Pro Xaa 1 5 10 15 <210> 55 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Phe (Cl) <400> 55 Arg Pro Arg Leu Ser Ala Arg Gly Pro Met Pro Xaa 1 5 10 <210> 56 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 11th position means Phe (Cl). <400> 56 Arg Pro Arg Leu Ser Ala Arg Gly Pro Xaa Xaa 1 5 10 <210> 57 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 12th position means Phe (Cl). <400> 57 Arg Pro Arg Leu Ser Ala Arg Gly Pro Xaa Pro Xaa 1 5 10 <210> 58 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means Cha. <400> 58 Arg Arg Gln Arg Pro Arg Leu Ser Ala Arg Gly Pro Cha 1 5 10 <210> 59 <211> 12 <212> PRT <2 13> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 12th position means Phe (Cl). <400> 59 Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Xaa 1 5 10 <210> 60 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Phe (Cl). <400> 60 Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Xaa 1 5 10 <210> 61 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 11th position means Cha. <400> 61 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Phe 1 5 10 <210> 62 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 13th position means Phe (Cl). <400> 62 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Xaa 1 5 10 <210> 63 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Cha. <400> 63 Arg Pro Arg Leu Ser Ala Arg Gly Pro Xaa Pro Phe 1 5 10 <210> 64 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 12th position means Phe (Cl). <400> 64 Arg Pro Arg Leu Phe Ala Arg Gly Pro Xaa Pro Xaa 1 5 10 <210> 65 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 12th position means Phe (Cl). <400> 65 Arg Pro Arg Leu Phe His Lys Gly Pro Xaa Pro Xaa 1 5 10 <210> 66 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa means Nle. <400> 66 Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly Pro Xaa Pro Tyr 1 5 10 15 <210> 67 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Cha. <400> 67 Arg Pro Arg Leu Phe His Lys Gly Pro Xaa Pro Phe 1 5 10 <210> 68 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Phe (Cl). <400> 68 Arg Pro Arg Leu Phe His Lys Gly Pro Met Pro Xaa 1 5 10 <210> 69 <211> 15 <212> PRT <213> Artificial Sequence <223><400> 69 Arg Arg Gln Arg Pro Arg Leu Ser Ala Arg Gly Pro Met Pro Phe 1 5 10 15 <210> 70 <211> 13 <212> PRT <213> Artificial Sequence <223><400> 70 Arg Arg Gln Arg Pro Arg Leu Ser Ala Arg Gly Pro Met 1 5 10 <210> 71 <211> 12 <212> PRT <213> Artific ial Sequence <223><400> 71 Arg Arg Gln Arg Pro Arg Leu Ser Ala Arg Gly Pro 1 5 10 <210> 72 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means Met (O) <400> 72 Arg Arg Gln Arg Pro Arg Leu Ser His Lys Gly Pro Xaa 1 5 10 <210> 73 <211> 15 <212> PRT <213> Artificial Sequence <223> Xaa means Lys (Arg-Arg) <400 > 73 Arg Arg Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 1 5 10 15 <210> 74 <211> 15 <212> PRT <213> Artificial Sequence <223><400> 74 Arg Arg Arg Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 1 5 10 15 <210> 75 <211> 15 <212> PRT <213> Artificial Sequence <223><400> 75 Arg Arg Lys Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 1 5 10 15 <210> 76 <211> 12 <212> PRT <213> Artificial Sequence <223><400> 76 Arg Pro Arg Leu Ser Ala Arg Gly Pro Met Pro Phe 1 5 10 <210> 77 <211> 15 <212> PRT <213> Artificial Sequence <223><400> 77 Arg Arg Ala Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 1 5 10 15 <210> 78 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 78 Xaa Arg Pro Arg Leu Ser Ala Arg Gly Pro Met Pro Phe 1 5 10 <210> 79 <211> 10 <212> PRT <213> Artificial Sequence <223><400> 79 Arg Pro Arg Leu Ser Ala Arg Gly Pro Met 1 5 10 <210> 80 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa means Phe (Cl) <400> 80 Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 81 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 81 Xaa Arg Pro Arg Leu Ser Ala Lys Gly Pro Met Pro Phe 1 5 10 <210> 82 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 82 Xaa Arg Pro Arg Leu Ser Arg Lys Gly Pro Met Pro Phe 1 5 10 <210> 83 <211> 12 <212> PRT <213> Artificial Sequence <223><400> 83 Arg Pro Arg Leu Phe Ala Arg Gly Pro Met Pro Phe 1 5 10 <210> 84 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Nal (2) <400> 84 Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Xaa 1 5 10 <210> 85 <211> 15 <212> PRT <213> Artificial Sequence <223><400> 85 Arg Arg Phe Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Phe 1 5 10 15 <210> 86 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 86 Xaa Arg Pro Arg Leu Ser His Arg Gly Pro Met Pro Phe 1 5 10 <210> 87 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 12th position means Phe (Cl) <400> 87 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 88 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa means Cha <400> 88 Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 89 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 11th position means Phe (Cl) <400> 89 Arg Pro Arg Leu Phe His Lys Gly Pro Xaa Xaa 1 5 10 <210> 90 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 90 Xaa Arg Pro Arg Leu Ser Leu Lys Gly Pro Met Pro Phe 1 5 10 <210> 91 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 13th position means Nal (2) <400> 91 Xaa Arg Pro Arg Leu Ser H is Lys Gly Pro Met Pro Xaa 1 5 10 <210> 92 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa means Nal (2) <400> 92 Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 93 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 93 Xaa Arg Pro Arg Leu Ser Arg Arg Gly Pro Met Pro Phe 1 5 10 <210> 94 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 94 Xaa Arg Pro Arg Leu Phe Arg Arg Gly Pro Met Pro Phe 1 5 10 <210> 95 <211> 13 <212 > PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 95 Xaa Arg Pro Arg Leu Ser Phe Lys Gly Pro Met Pro Phe 1 5 10 <210> 96 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 96 Xaa Arg Pro Arg Leu Phe His Lys Gly Pro Met Pro Phe 1 5 10 <210> 97 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 1st position means pGlu, Xaa on the 12th position means Cha <400> 97 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 98 <211> 12 <212> PRT <213> Artificial Sequence <223> Xa a on the 1st position means pGlu, Xaa on the 12th position means Nal (2) <400> 98 Xaa Arg Pro Arg Leu Ser His Lys Gly Pro Met Xaa 1 5 10 <210> 99 <211> 11 <212> PRT <213> Artificial Sequence <223><400> 99 Arg Pro Arg Leu Phe Ala Arg Gly Pro Met Phe 1 5 10 <210> 100 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400 > 100 Xaa Arg Pro Arg Leu Ser His Phe Gly Pro Met Pro Phe 1 5 10 <210> 101 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Cha <400> 101 Arg Pro Arg Leu Ser His Lys Gly Pro Met Pro Xaa 1 5 10 <210> 102 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa means pGlu <400> 102 Xaa Arg Pro Arg Leu Ser His Leu Gly Pro Met Pro Phe 1 5 10 <210> 103 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means NMe ₂ <400> 103 Arg Arg Gln Arg Pro Arg Leu Ser Ala Arg Gly Xaa 1 5 10 <210> 104 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Mor <400> 104 Arg Arg Gln Arg Pro Arg Leu Ser Ala Arg Gly Xaa 1 5 10 <210> 105 <211> 12 <212> PRT <21 3> Artificial Sequence <223> Xaa means Phe (Cl) <400> 105 Arg Pro Arg Leu Ser Ala Arg Gly Pro Ala Pro Xaa 1 5 10 <210> 106 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa means Phe (Cl) <400> 106 Arg Pro Arg Leu Ser Ala Arg Gly Gly Met Pro Xaa 1 5 10 <210> 107 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 9th position means N-MeAla, Xaa on the 12th position me ans Phe (Cl) <400> 107 Arg Pro Arg Leu Ser Ala Arg Gly Xaa Met Pro Xaa 1 5 10 <210> 108 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 12th position means Phe (Cl) <400> 108 Arg Pro Arg Leu Ser His Ala Gly Pro Xaa Pro Xaa 1 5 10 <210> 109 <211 > 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 12th position means Phe (Cl) <400> 109 Arg Pro Arg Ala Ser His Lys Gly Pro Xaa Pro Xaa 1 5 10 <210> 110 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 12th position means Phe (Cl) <400> 110 Arg Pro Ala Leu Ser His Lys Gly Pro Xaa Pro Xaa 1 5 10 <210> 111 <211> 11 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 11th position means Pyn <400> 111 Arg Pro Arg Leu Ser Ala Arg Gly Pro Xaa Xaa 1 5 10 <210> 112 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 10th position means Cha, Xaa on the 12th position means Pyn <400> 112 Arg Pro Arg Leu Ser Ala Arg Gly Pro Xaa Pro Xaa 1 5 10 <210> 113 <211> 13 <212> PRT <213> Artificial Sequence <223> Xaa on the 13th position means Cha <400> 113 Arg Arg Gln Arg Pro Arg Leu Ser Ala Arg Gly Gly Xaa 1 5 10 <210> 114 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 3rd position means Lys ( Me) ₂ <400> 114 Arg Pro Xaa Leu Ser Ala Arg Gly Pro Met Pro Phe 1 5 10 <210> 115 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 7th position means Lys ( Me) ₂ <400> 115 Arg Pro Arg Leu Ser Ala Xaa Gly Pro Met Pro Phe 1 5 10 <210> 116 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 6th position means Dap, Xaa on the 10th position means Cha, Xaa on the 12th position means Phe (Cl) <400> 116 Arg Pro Arg Leu Ser Xaa Arg Gly Pro Xaa Pro Xaa 1 5 10 <210> 117 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 6th position means Dap (Ac), Xaa on the 10th position me ans Cha, Xaa on the 12th position means Phe (Cl) <400> 117 Arg Pro Arg Leu Ser Xaa Arg Gly Pro Xaa Pro Xaa 1 5 10 <210> 118 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 6th position means Dap (C ₆ ), Xaa on the 10th position me ans Cha, Xaa on the 12th position means Phe (Cl) <400> 118 Arg Pro Arg Leu Ser Xaa Arg Gly Pro Xaa Pro Xaa 1 5 10 <210> 119 <211> 12 <212> PRT <213> Artificial Sequence <223> Xaa on the 6th position means Dap (Adi), Xaa on the 10th position m eans Cha, Xaa on the 12th position means Phe (Cl) <400> 119 Arg Pro Arg Leu Ser Xaa Arg Gly Pro Xaa Pro Xaa 1 5 10

[Brief description of drawings]

FIG. 1 shows changes in blood ACTH, LH, TSH, FSH, Prolactin, oxytocin, and Arg-Vasopressin concentrations 10 minutes after administration of apelin into the rat third ventricle.
The vehicle on the horizontal axis shows the case where phosphate buffered saline was infused, and Apelin shows the case where rat apelin was infused. The vertical axis represents the hormone concentration in plasma, and ACTH
Indicates corticotropin, LH indicates luteinizing hormone, TSH indicates thyroid stimulating hormone, FSH indicates follicle stimulating hormone, PRL indicates prolactin, AVP indicates arginine-vasopressin, and Oxytocin indicates oxytocin. Values are mean ± standard deviation (mean ± SE
M) (n = 4). * Indicates that the P value is 0.05 or less as compared with the phosphate buffered saline administration group.

FIG. 2 shows time-dependent changes in blood ACTH concentration when apelin was administered to the third ventricle of the rat. The horizontal axis represents the time (minutes) after administration. The vertical axis represents the concentration of adrenocorticotropic hormone (ACTH) in plasma. A vehicle of Δ indicates a case where phosphate buffered saline is injected. ○ Ape
Lin 3 nmol shows the case of injecting rat apelin 3 nmol. Apelin 10 nmol of ● indicates the case where 10 nmol of rat apelin was injected. Values are mean ± standard deviation (mean ± SEM) (n = 6-7)
Indicates. * Indicates P compared to phosphate-buffered saline administration group
The value is 0.05 or less. ** indicates that the P value is 0.01 or less as compared with the phosphate buffered saline administration group.

FIG. 3 shows time-course changes in blood oxytocin concentration when apelin was administered to the third ventricle of the rat. The horizontal axis represents the time (minutes) after administration. The vertical axis represents the oxytocin concentration in plasma. A vehicle of Δ indicates a case where phosphate buffered saline is injected. ○ Apelin 3nmo
1 shows the case where 3 nmol of rat apelin was injected.
● Apelin 10 nmol is rat apelin 10
The case where nmol is injected is shown. Values represent mean ± standard deviation (mean ± SEM) (n = 6-7). ** indicates that the P value is 0.01 or less as compared with the phosphate buffered saline administration group.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 48/00 A61P 1/00 4C086 A61P 1/00 1/04 4H045 1/04 3/00 3/00 3 / 04 3/04 5/06 5/06 5/10 5/10 5/38 5/38 5/40 5/40 5/44 5/44 13/12 13/12 15/00 15/00 15/04 15/04 15/06 15/06 15/08 15/08 15/14 15/14 25/18 25/18 25/24 25/24 29/00 29/00 35/00 35/00 43/00 111 43 / 00 111 C12Q 1/68 A C12Q 1/68 G01N 33/15 Z G01N 33/15 33/50 Z 33/50 33/53 D 33/53 M 33/566 33/566 C07K 14/47 // C07K 14 / 47 A61K 37/24 ZNA C12N 15/09 C12N 15/00 A (72) Inventor Shuji Hinuma 1-7 Kasuga, Tsukuba, Ibaraki Prefecture 9-9 Takeda Kasuga Heights 1402 F-term (reference) 2G045 AA34 AA35 AA40 BA11 DA12 DA13 DA14 DA36 FB02 F B03 4B024 AA01 AA11 BA63 BA80 CA02 GA11 HA08 4B063 QA18 QQ08 QQ42 QQ61 QR32 QR55 QR72 QR77 QS28 QS34 QS36 4C084 AA13 AA17 BA01 BA08 BA18 CA18 CA53 CA59 C59 DB59 DB01 DB14 NA14 ZA08 ZA12B81Z68C12 BB07 CC02 CC07 CC08 CC21 DD23 DD62 DD63 EE01 4C086 AA01 AA02 EA16 MA01 MA04 NA14 ZA08 ZA12 ZA18 ZA66 ZA68 ZA70 ZA81 ZB11 ZB26 ZC04 ZC08 ZC41 ZC42 4H045 AA10 AA30 BA09 CA40 FA74 EA20 EA20 EA20 EA20

Claims (102)

[Claims] 1. A corticotropin secretory regulator comprising apelin, a modified form thereof, an amide thereof, an ester thereof or a salt thereof. 2. Hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease, adrenal insufficiency, comprising apelin, its modified form or its amide or its ester or its salt. A prophylactic / therapeutic agent for pain or obesity. 3. An oxytocin secretion regulator comprising apelin, its modified form, its amide, its ester or its salt. 4. A labor inducer comprising apelin, its modified form or its amide or its ester or its salt, weak labor, relaxation bleeding, uterine restoration failure, lactation failure, post-traumatic stress syndrome or milk stasis. Prevention of
A therapeutic agent or an adjunct agent before and after placental delivery, cesarean section, miscarriage or artificial abortion. 5. D containing a DNA encoding apelin
An adrenocorticotropic hormone secretory regulator comprising NA. 6. D containing a DNA encoding apelin
A prophylactic / therapeutic agent for hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease, adrenal insufficiency, pain or obesity, which comprises NA. 7. D containing a DNA encoding apelin
An oxytocin secretion regulator comprising NA. 8. D containing a DNA encoding apelin
A labor inducer containing NA, weak labor, relaxation bleeding,
A preventive / therapeutic agent for uterine retrograde failure, lactation failure, post-traumatic stress syndrome or lactation, or an adjunct agent before and after placental delivery, cesarean section, miscarriage or artificial abortion. 9. An adrenocorticotropic hormone secretory regulator comprising an antibody against apelin, its modified form, its amide, its ester, or its salt. 10. Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosterone, which comprises an antibody against apelin, its modified form or its amide or its ester or its salt. Agent for the treatment of psoriasis, primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa, gastroduodenal ulcer or irritable bowel syndrome. 11. An oxytocin secretion regulator comprising an antibody against apelin, its modified form, its amide, its ester, or its salt. 12. Overcompetent labor, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth, which comprises an antibody against apelin, its modified form, its amide or its ester, or its salt. Or a prophylactic / therapeutic agent for Prader-Willi syndrome. 13. A corticotropin secretion regulator comprising antisense DNA against DNA containing apelin-encoding DNA. 14. A Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism, primary comprising antisense DNA against DNA containing apelin-encoding DNA. Cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression,
Prophylactic / therapeutic agent for anorexia nervosa, gastric / duodenal ulcer, or irritable bowel syndrome. 15. An oxytocin secretion regulator comprising antisense DNA against DNA containing apelin-encoding DNA. 16. Overcompensation, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prader- comprising antisense DNA against DNA containing DNA encoding apelin. Agent for the prevention and treatment of Willi syndrome. 17. A diagnostic agent for abnormal adrenocorticotropic hormone secretion, comprising an antibody against apelin, its modified form, its amide, its ester, or its salt. 18. A diagnostic agent for abnormal oxytocin secretion, which comprises an antibody against apelin, its modified form, its amide, its ester, or its salt. 19. Hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease, adrenal function comprising an antibody against apelin, its modified form or its amide or its ester or its salt. Dysfunction, pain, obesity, weak labor, loose bleeding, uterine restoration failure, lactation failure, post-traumatic stress syndrome, milk stasis, Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor, aldosterone production Tumor, aldosteronism, primary cortisol resistance, congenital adrenal deficiency, stress, depression, anorexia nervosa, gastroduodenal ulcer, irritable bowel syndrome, hypertonic pain, ankylosing uterine contraction, fetal distress, Diagnostic agent for uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-Willi syndrome. 20. A diagnostic agent for abnormal adrenocorticotropic hormone secretion, which comprises a DNA containing a DNA encoding apelin. 21. A diagnostic agent for abnormal oxytocin secretion, which comprises a DNA containing a DNA encoding apelin. 22. Hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticalism, Addison's disease, adrenal insufficiency, pain, obesity, weakness comprising DNA containing DNA encoding apelin Labor, laxative bleeding, uterine retrograde failure, lactation insufficiency, posttraumatic stress syndrome, milk stasis, Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor,
Aldosterone-producing tumor, aldosteronism, primary cortisol resistance, congenital adrenal deficiency, stress,
Depression, anorexia nervosa, gastroduodenal ulcer, irritable bowel syndrome, overwork pain, ankylosing uterus, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-Willi
Diagnostic agent for syndrome. 23. The agent according to claim 1, wherein apelin is a peptide containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 1. 24. Apelin has SEQ ID NO: 5 and SEQ ID NO:
7. A peptide containing a partial sequence of the amino acid sequence represented by SEQ ID NO: 9 or SEQ ID NO: 11.
22. The agent of 22. 25. Apelin is (a) SEQ ID NO: 5, 7, 9
Or the 6th to 7th of the amino acid sequence represented by 11
Peptide having the 7th amino acid sequence, (b) Peptide having the 40th to 77th amino acid sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (c) SEQ ID NO: 5 , A peptide having the 42nd to 77th amino acid sequence of the amino acid sequence represented by 7, 9 or 11, (d) the 47th position of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11. From 77th
A peptide having the amino acid sequence of position (e), a peptide having the 61st to 77th amino acid sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (f) SEQ ID NO: 5, A peptide having the 65th to 77th amino acid sequence of the amino acid sequence represented by 7, 9 or 11 or its N-terminal amino acid (Gl
(n) a peptide having an amino acid sequence that is pyroglutamine-oxidized, (g) a peptide having the 1st to 25th amino acid sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (h) 6th to 25th amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11
A peptide having the amino acid sequence of position (i), a peptide having the 42nd to 64th amino acid sequence of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (j) SEQ ID NO: 5, A peptide having the 61st to 64th amino acid sequence of the amino acid sequence represented by 7, 9 or 11, (k) from the 43rd position of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 77th
A peptide having the amino acid sequence of (1), (l) a peptide having the amino acid sequence of 41st to 77th of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (m) SEQ ID NO: 5, A peptide having the 66th to 77th amino acid sequence of the amino acid sequence represented by 7, 9 or 11, (n) from the 67th amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 77th
A peptide having the amino acid sequence of position (o), a peptide having the amino acid sequence from the 64th position to the 77th position of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11, (p) SEQ ID NO: 5, A peptide having the 63rd to 77th amino acid sequence of the amino acid sequence represented by 7, 9 or 11, (q) from the 65th amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11 76th
23. The peptide having the amino acid sequence of the third position or (r) the peptide having the amino acid sequence of the 65th to 75th positions of the amino acid sequence represented by SEQ ID NO: 5, 7, 9 or 11. 23. Agent. 26. Apelin has SEQ ID NO: 5 and SEQ ID NO:
23. The agent according to claim 1, which is a peptide containing the 65th to 77th amino acid sequences of the amino acid sequence represented by 7, SEQ ID NO: 9 or SEQ ID NO: 11. 27. Apelin is pGlu Arg Pro Arg Leu Ser H
23. The agent according to claim 1, which is a peptide containing an amino acid sequence represented by is Lys Gly Pro Met ProPhe. 28. Apelin has SEQ ID NO: 5 and SEQ ID NO:
23. The agent according to claims 1 to 22, which is a peptide containing the 42nd to 77th amino acid sequences of the amino acid sequence represented by 7, SEQ ID NO: 9 or SEQ ID NO: 11. 29. A modified form of apelin has the formula X ¹ -Arg-Pro-Arg-X ² -Ser-His-X ³ -Gly-Pro-X ⁴ -X ⁵ (I) [wherein X ¹ is hydrogen. An amino acid or an amino acid residue or a peptide chain consisting of 1 to 25 amino acids, each of which may be the same or different and whose side chain is substituted,
² represents a neutral amino acid residue whose side chain may be substituted, X ³ represents a neutral amino acid residue whose side chain may be substituted, and an aromatic amino acid residue whose side chain may be substituted. A basic amino acid residue whose group or side chain may be substituted, X ⁴ represents a neutral or aromatic amino acid residue whose bond or side chain may be substituted, and X ⁵ represents a side chain An amino acid residue which may be substituted or an amino acid derivative in which the C-terminal carboxyl group thereof is reduced to a hydroxymethyl group or a formyl group, an amino acid residue which may be substituted a hydroxyl group or a side chain and a side chain are substituted A dipeptide chain having an optionally attached amino acid residue or a C-terminal carboxyl group thereof is reduced to a hydroxymethyl group or a formyl group to represent a peptide derivative represented by the formula -Arg-Pro- Arg-, -Ser-His-
Alternatively, the side chain of each amino acid residue in -Gly-Pro- may be substituted. However, X ² represents Leu, X ³ represents Lys, X ⁴ represents Met, and X ⁵ represents Pro or Pro-Phe, and -Arg-Pro in the formula.
-Arg- is an unsubstituted -Arg-Pro-Arg-,
-Ser-His- is an unsubstituted -Ser-His-
And -Gly-Pro- is an unsubstituted -Gly-Pro-
Except when ] The agent of Claims 1-4, 9-12, and 17-19 which is a peptide represented by these. 30. A modified form of apelin has the formula P1-Arg-Pro-Arg-Leu-Phe-P2-P3-Gly-Pro-P4-P5 (II) [wherein P1 is a hydrogen atom or the same or different, respectively] An amino acid residue or a peptide chain consisting of 1 to 25 amino acids whose side chain may be substituted,
2 represents a neutral amino acid residue whose side chain may be substituted or a basic amino acid residue whose side chain may be substituted, and P3 is a neutral amino acid residue whose side chain may be substituted. , A side chain may be substituted with an aromatic amino acid residue or a side chain may be substituted with a basic amino acid residue, P4 is a bond or a neutral side chain may be substituted or An aromatic amino acid residue is shown. P5 is an amino acid residue whose side chain may be substituted or an amino acid derivative in which its C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group, or a hydroxyl group or side chain is substituted. A dipeptide chain consisting of an amino acid residue which may be substituted with an amino acid residue whose side chain may be substituted or its C-terminal carboxyl group is reduced to a hydroxymethyl group or a formyl group. Shows the peptide derivative, the side chain of -Arg-Pro-Arg-Leu-Phe- or -Gly-Pro- each amino acid residue in the middle in the formula may be substituted. ] It is a peptide represented by these.
~ 4, 9-12 and 17-19 described agents. 31. A modified form of apelin has the formula Q1-Arg-Pro-Arg-Leu-Ser-Ala-Q2-Gly-Q5-Q3-Q4 (III) [wherein Q1 is a hydrogen atom or the same or different, respectively] An amino acid residue or a peptide chain consisting of 1 to 25 amino acids whose side chain may be substituted,
2 represents a neutral amino acid residue whose side chain may be substituted, an aromatic amino acid residue whose side chain may be substituted, or a basic amino acid residue whose side chain may be substituted,
Q3 represents a neutral or aromatic amino acid residue whose bond or side chain may be substituted, Q4 represents an amino acid residue whose side chain may be substituted or its C-terminal carboxyl group is a hydroxymethyl group or A formyl group-reduced amino acid derivative, a dipeptide chain formed by binding an amino acid residue which may be substituted with a hydroxyl group or a side chain and an amino acid residue which may be substituted with a side chain, or its C-terminal carboxyl group is Shows a peptide derivative reduced to a hydroxymethyl group or a formyl group, Q5 represents a neutral amino acid residue whose side chain may be substituted, and is -Arg-Pro-Arg-Leu-Ser-Ala in the formula.
The side chain of each amino acid residue in-may be substituted. ] The peptide represented by these, its ester, its amide, or its salt, 1-4, 9-1.
2 and 17-19. 32. A modified form of apelin is represented by the formula R1-Arg-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-R2-Pro-R3 (IV) [wherein R1 is a hydrogen atom or respectively. An amino acid residue or a peptide chain consisting of 1 to 25 amino acids, which may be the same or different and whose side chains are substituted,
2 represents Cha which may be substituted, Met which may be substituted or Nle which may be substituted, and R
3 is optionally substituted Phe (Cl), optionally substituted Phe, optionally substituted Nal
(2) represents an optionally substituted Cha or an optionally substituted Tyr, and represents -Arg-Pro-A in the formula.
rg-Leu-Ser-His-Lys-Gly-Pr
The side chain of each amino acid residue in o or -Pro- may be substituted. ] It is a peptide represented by these, its ester, its amide, or its salt.
~ 4, 9-12 and 17-19 described agents. 33. Apelin, a modified product thereof, an amide thereof, an ester thereof, a salt thereof and / or
A method for screening a corticotropin secretion-regulating substance, which comprises using an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3, a partial peptide thereof or a salt thereof. 34. A DNA containing a DNA encoding apelin and / or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a partial peptide thereof is encoded. A method for screening an adrenocorticotrophic hormone secretion regulator, which comprises using DNA. 35. Apelin, a modified form thereof, an amide thereof, an ester thereof, a salt thereof and / or
A method for screening an oxytocin secretion regulator, which comprises using an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3, a partial peptide thereof or a salt thereof. 36. A DNA containing a DNA encoding apelin and / or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a partial peptide thereof is encoded. A method for screening an oxytocin secretion regulator, which comprises using DNA. 37. An apelin receptor, wherein the substance contains apelin, a modified product thereof, an amide thereof, an ester thereof or a salt thereof, and an amino acid sequence which is the same or substantially the same as the amino acid sequence represented by SEQ ID NO: 3. 37. The screening method according to claim 33, which is a substance that changes the binding property to the partial peptide or the salt thereof. 38. A substance which regulates the expression of apelin receptor or a salt thereof and / or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a salt thereof. The screening method according to any one of claims 33 to 36. 39. Apelin, a modified product thereof, an amide thereof, an ester thereof, a salt thereof and / or
For screening an adrenocorticotrophic hormone secretory regulator characterized by containing an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3, its partial peptide or its salt kit. 40. A DNA containing a DNA encoding apelin and / or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a partial peptide thereof is encoded. A kit for screening an adrenocorticotropic hormone secretion regulator, which comprises DNA containing DNA. 41. Apelin, a modified product thereof, an amide thereof, an ester thereof, a salt thereof and / or
A kit for screening an oxytocin secretion-regulating substance, which comprises an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3, a partial peptide thereof or a salt thereof. 42. A DNA containing a DNA encoding apelin and / or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a partial peptide thereof is encoded. A kit for screening an oxytocin secretion regulator, which comprises DNA containing DNA. 43. An adrenocorticotropic hormone secretory substance obtained by using the screening method according to claim 33 or 34 or the screening kit according to claim 39 or 40. 44. An oxytocin secretion regulator obtained using the screening method according to claim 33 or 34 or the screening kit according to claim 39 or 40. 45. An adrenocorticotropic hormone secretory regulator comprising the substance according to claim 43. 46. Hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease, adrenal insufficiency, comprising the substance according to claim 43.
A prophylactic / therapeutic agent for pain or obesity. 47. Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism, primary cortisol resistance, which comprises the substance according to claim 43.
A prophylactic / therapeutic agent for congenital adrenal enzyme deficiency, Addison's disease, stress, depression, anorexia nervosa, gastric / duodenal ulcer or irritable bowel syndrome. 48. An oxytocin secretion regulator comprising the substance according to claim 44. 49. A prophylactic / therapeutic agent for labor delivery inducer, weak labor, relaxation bleeding, uterine restoration failure, lactation deficiency, posttraumatic stress syndrome or milk stasis, or placenta containing the substance according to claim 44. Adjuvant before and after delivery, cesarean section, miscarriage or artificial abortion. 50. A prophylactic / therapeutic agent for hypertonic labor, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-Willi syndrome, which comprises the substance according to claim 44. .. 51. (1) An apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3, with apelin, its modified form or its amide or its ester or its salt, A substance that changes the binding property to a partial peptide or a salt thereof, or (2) apelin or a salt thereof and / or an apelin containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3. A corticotropin secretory regulator comprising a substance that regulates the expression of a receptor or a salt thereof. 52. (1) An apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 with apelin, its modified form or its amide or its ester or its salt, A substance that changes the binding property to a partial peptide or a salt thereof, or (2) apelin or a salt thereof and / or an apelin containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3. An oxytocin secretion regulator comprising a substance that regulates the expression of a receptor or a salt thereof. 53. An agonist for an apelin receptor or its salt, which contains the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3, or (2) apelin or its salt and / or SEQ ID NO: : A substance having an activity of promoting the expression of an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by 3, or a salt thereof, hypoaldosteronism, hypocortisolemia, A preventive / therapeutic agent for secondary and chronic hypoadrenocorticism, Addison's disease, adrenal insufficiency, pain or obesity. 54. An agonist for an apelin receptor or a salt thereof, which contains the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3, or (2) apelin or a salt thereof and / or SEQ ID NO: : A substance having an activity of promoting the expression of an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by 3: or a salt thereof, a parturition-inducing agent, weak labor, and relaxation bleeding , A preventive / therapeutic agent for uterine retrograde failure, lactation failure, post-traumatic stress syndrome or milk stasis, or an adjunct agent before and after placental delivery, cesarean section, miscarriage or artificial abortion. 55. An antagonist to an apelin receptor or a salt thereof, which contains the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3.
Or (2) apelin or its salt and / or
Cushing's disease, Cushing's syndrome, adrenal gland comprising a substance having an activity of inhibiting the expression of an apelin receptor or its salt containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3. Corticotropin-producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism, primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa, gastroduodenal ulcer or Agent for the prevention and treatment of irritable bowel syndrome. 56. An antagonist to an apelin receptor or a salt thereof, which contains the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3.
Or (2) apelin or its salt and / or
A hypertonic labor, ankylosing uterus comprising a substance having an activity of inhibiting the expression of an apelin receptor or a salt thereof containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3. Contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-Wil
Preventive and therapeutic agent for li syndrome. 57. A method for regulating adrenocorticotropic hormone secretion, which comprises administering an effective amount of apelin, its modified form, its amide, its ester, or its salt to a mammal. 58. Hypoaldosteronism, hypocortisolemia, secondary and chronic adrenal glands, characterized in that an effective amount of apelin, its modified form or its amide or its ester or its salt is administered to a mammal. A method for preventing / treating hypocortical dysfunction, Addison's disease, adrenal insufficiency, pain or obesity. 59. A method for regulating oxytocin secretion, which comprises administering an effective amount of apelin, its modified form, its amide, its ester, or its salt to a mammal. 60. A method for inducing parturition, weak labor, loose bleeding, uterine restoration failure, which comprises administering an effective amount of apelin, its modified form, its amide or its ester, or its salt to a mammal. Preventive and therapeutic methods for lactation insufficiency, post-traumatic stress syndrome or lactation, or assistance before and after placental delivery, cesarean section, miscarriage or artificial abortion. 61. An apelin receptor containing a DNA encoding apelin or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a partial peptide thereof for mammals. A method for regulating adrenocorticotropic hormone secretion, which comprises administering an effective amount of DNA containing DNA encoding 62. An apelin receptor or a partial peptide thereof, which comprises a DNA containing a DNA encoding apelin or an amino acid sequence which is the same or substantially the same as the amino acid sequence represented by SEQ ID NO: 3 for mammals. Hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenal cortex, characterized by administering an effective amount of DNA containing DNA encoding
A method for preventing or treating Addison's disease, adrenal insufficiency, pain or obesity. 63. An apelin receptor containing a DNA encoding apelin or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a partial peptide thereof for mammals. A method for regulating oxytocin secretion, which comprises administering an effective amount of DNA containing DNA encoding 64. An apelin receptor or a partial peptide thereof, which comprises a DNA containing a DNA encoding apelin or an amino acid sequence which is the same or substantially the same as the amino acid sequence represented by SEQ ID NO: 3 for mammals. For inducing parturition, faint labor pain, laxative bleeding, uterine restoration failure, lactation failure, posttraumatic stress syndrome or milk stasis, which comprises administering an effective amount of DNA containing DNA encoding , Or before and after placental delivery, Caesarean section, miscarriage or artificial abortion assistance. [65] A method for regulating adrenocorticotropic hormone secretion, which comprises administering an effective amount of the substance according to [43] to a mammal. 66. A hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease, which comprises administering to a mammal an effective amount of the substance according to claim 43. A method for preventing or treating adrenal insufficiency, pain or obesity. 67. A Cushing's disease, which comprises administering an effective amount of the substance according to claim 43 to a mammal,
Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism, primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa, stomach -Method for preventing and / or treating duodenal ulcer or irritable bowel syndrome. 68. A method for regulating oxytocin secretion, which comprises administering an effective amount of the substance according to claim 44 to a mammal. 69. A method for inducing labor, which comprises administering an effective amount of the substance according to claim 44 to a mammal.
A method for the prevention and treatment of weak labor, flaccid bleeding, uterine restoration failure, lactation failure, post-traumatic stress syndrome or milk stasis, or an adjunct method before and after placental delivery, cesarean section, miscarriage or artificial abortion. 70. An excessive amount of the substance according to claim 44, which is administered to a mammal, excessive labor pain, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea. How to prevent and treat preterm birth or Prader-Willi syndrome. 71. An amino acid sequence which is the same or substantially the same as the amino acid sequence represented by SEQ ID NO: 3 with (1) apelin, its modified form or its amide or its ester or its salt, with respect to mammals. A substance containing apelin receptor, a partial peptide or a salt thereof that changes the binding property, or (2) apelin or a salt thereof and / or the same or substantially the same as the amino acid sequence represented by SEQ ID NO: 3. A method for regulating adrenocorticotropic hormone secretion, which comprises administering an effective amount of a substance that regulates the expression of an apelin receptor containing an amino acid sequence or a salt thereof. 72. An amino acid sequence which is the same or substantially the same as the amino acid sequence represented by SEQ ID NO: 3 with (1) apelin, its modified form or its amide or its ester or its salt, for mammals. A substance containing apelin receptor, a partial peptide or a salt thereof that changes the binding property, or (2) apelin or a salt thereof and / or the same or substantially the same as the amino acid sequence represented by SEQ ID NO: 3. A method for regulating oxytocin secretion, which comprises administering an effective amount of a substance that regulates the expression of an apelin receptor containing an amino acid sequence or a salt thereof. 73. An apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 with respect to mammals, an agonist for the partial peptide or a salt thereof, or (2). Administering an effective amount of an apelin receptor or a salt thereof and / or a substance having an activity of promoting the expression of an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a salt thereof A method for preventing / treating hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenal cortex, Addison's disease, adrenal insufficiency, pain or obesity. 74. For a mammal, (1) SEQ ID NO: 3
An apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by, an agonist for a partial peptide or a salt thereof, or (2)
Apelin or its salt and / or SEQ ID NO:
The method for inducing parturition, which comprises administering an effective amount of a substance having an activity of promoting the expression of an apelin receptor or a salt thereof containing the same or substantially the same amino acid sequence as the amino acid sequence represented by 3. Labor pain, relaxation bleeding, uterine restoration failure, lactation failure, post-traumatic stress syndrome or milk stasis prevention or treatment method, or before and after placental delivery, cesarean section, miscarriage or assisted abortion. 75. With respect to mammals, (1) SEQ ID NO: 3
An apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by, an antagonist to a partial peptide or a salt thereof, or (2) apelin or a salt thereof and / or represented by SEQ ID NO: 3. Cushing's disease, Cushing's syndrome, adrenocortical stimulation, which comprises administering an effective amount of a substance having an activity of inhibiting the expression of an apelin receptor or its salt containing the same or substantially the same amino acid sequence as the amino acid sequence Hormone-producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism,
Primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa, stomach /
A method for preventing and / or treating duodenal ulcer or irritable bowel syndrome. 76. For a mammal, (1) SEQ ID NO: 3
An apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by, an antagonist to a partial peptide thereof or a salt thereof, or (2) apelin or a salt thereof and / or represented by SEQ ID NO: 3. Overwork labor, tonic uterine contraction, characterized by administering an effective amount of a substance having the activity of inhibiting the expression of an apelin receptor or its salt containing the same or substantially the same amino acid sequence as the amino acid sequence,
Methods for the prevention and treatment of fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-Willi syndrome. 77. Use of apelin, a modified form thereof, an amide thereof, an ester thereof, or a salt thereof for producing a corticotropin secretion regulator. 78. Apelin for producing a prophylactic / therapeutic agent for hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease, adrenal insufficiency, pain or obesity, or a modified form thereof. Use of the amide or ester thereof or salt thereof. 79. Use of apelin, its modified form, its amide or its ester, or its salt for producing an oxytocin secretion regulator. 80. A labor inducement agent, weak labor, relaxation bleeding, uterine restoration failure, lactation failure, post-traumatic stress syndrome or prevention / treatment of milk stasis, or placental delivery, cesarean section, miscarriage or artificial pregnancy. Use of apelin, its modified form or its amide or its ester or its salt for producing an auxiliary agent for abortion. 81. A D containing a DNA encoding apelin for producing a corticotropin secretion regulator.
DN encoding NA or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a partial peptide thereof
Use of DNA containing A. 82. A DNA encoding an apelin for producing a prophylactic / therapeutic agent for hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocortical function, Addison's disease, adrenal insufficiency, pain or obesity. D encoding the contained DNA or apelin receptor or its partial peptide containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3
Use of DNA containing NA. 83. A DNA containing DNA encoding apelin for producing an oxytocin secretion regulator, or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3, or Use of DNA containing DNA encoding the partial peptide. 84. A prophylactic / therapeutic agent for labor delivery, weak labor, flaccid bleeding, uterine retrograde failure, lactation insufficiency, posttraumatic stress syndrome or milk stasis, or before / after placental delivery, cesarean section, miscarriage or artificial pregnancy. D containing DNA encoding apelin for the production of abortion aids
DN encoding NA or an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 or a partial peptide thereof
Use of DNA containing A. 85. Use of the substance according to claim 43 for producing a corticotropin secretion regulator. 86. The substance according to claim 43 for producing a prophylactic / therapeutic agent for hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocortical function, Addison's disease, adrenal insufficiency, pain or obesity. Use of. 87. Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism, primary cortisol resistance, congenital adrenal deficiency, Addison's disease,
44. Use of the substance according to claim 43 for producing a prophylactic / therapeutic agent for stress, depression, anorexia nervosa, gastroduodenal ulcer or irritable bowel syndrome. 88. Use of the substance according to claim 44 for producing an oxytocin secretion regulator. 89. A delivery inducer, a weak labor, relaxation bleeding, uterine retrograde failure, lactation insufficiency, a preventive / therapeutic agent for posttraumatic stress syndrome or lactation, or before and after placental delivery, cesarean section, miscarriage or artificial pregnancy. Use of a substance according to claim 44 for the production of an abortion supplement. 90. Hypertonic labor, ankylosing uterine contraction, fetal distress,
Uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-W
45. A method for producing a prophylactic / therapeutic agent for illi syndrome.
Use of the listed substances. 91. (1) Apelin, a modified form thereof, an amide thereof, an ester thereof or a salt thereof for producing a corticotropin secretion regulator, and SEQ ID NO: 3
Apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by the following, a substance that changes the binding property to its partial peptide or its salt, or (2) apelin or its salt and / or sequence Use of a substance that regulates the expression of an apelin receptor or a salt thereof, which contains the same or substantially the same amino acid sequence as the amino acid sequence represented by No. 3. 92. The same or substantially the same as the amino acid sequence represented by SEQ ID NO: 3 with (1) apelin, its modified form or its amide or its ester or its salt for producing an oxytocin secretion regulator. Apelin receptor containing an amino acid sequence, a substance that changes the binding property to a partial peptide or a salt thereof, or (2)
Apelin or its salt and / or SEQ ID NO:
Use of a substance that regulates the expression of an apelin receptor or a salt thereof containing the same or substantially the same amino acid sequence as the amino acid sequence represented by 3. 93. (1) SEQ ID NO: for producing a prophylactic / therapeutic agent for hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocortical function, Addison's disease, adrenal insufficiency, pain or obesity. An apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by 3, an agonist for a partial peptide or a salt thereof, or (2) apelin or a salt thereof and / or represented by SEQ ID NO: 3 Use of a substance having an activity of promoting the expression of an apelin receptor or a salt thereof containing the same or substantially the same amino acid sequence as the amino acid sequence described above. 94. A delivery inducer, weak labor, relaxation bleeding, uterine restoration failure, lactation failure, post-traumatic stress syndrome or prevention / treatment of milk stasis, or placental delivery, cesarean section, miscarriage or artificial pregnancy. (1) an apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 for producing an adjuvant at abortion, an agonist for a partial peptide or a salt thereof, or ( 2) Use of a substance having an activity of promoting the expression of an apelin receptor or a salt thereof and / or a salt thereof, which contains the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3. 95. Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor, aldosterone-producing tumor, aldosteronism, primary cortisol resistance, congenital adrenal enzyme deficiency, Addison's disease,
(1) The same or substantially the same amino acid sequence represented by SEQ ID NO: 3 for producing a prophylactic / therapeutic agent for stress, depression, anorexia nervosa, gastroduodenal ulcer or irritable bowel syndrome An apelin receptor containing an amino acid sequence, an antagonist to a partial peptide or a salt thereof, or (2) an apelin or a salt thereof and / or an amino acid sequence identical or substantially identical to the amino acid sequence represented by SEQ ID NO: 3 Use of a substance having an activity of inhibiting the expression of the contained apelin receptor or a salt thereof. 96. Hypertonic labor, ankylosing uterine contraction, fetal distress,
Uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prader-W
(1) An apelin receptor containing the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3 for producing a prophylactic / therapeutic agent for illi syndrome, an antagonist to a partial peptide or a salt thereof,
Or (2) apelin or its salt and / or
Use of a substance having an activity of inhibiting the expression of an apelin receptor or a salt thereof, which contains the same or substantially the same amino acid sequence as the amino acid sequence represented by SEQ ID NO: 3. 97. A method for regulating adrenocorticotropic hormone secretion, which comprises regulating the activity of mammalian apelin receptor. 98. A method for regulating oxytocin secretion, which comprises regulating the activity of mammalian apelin receptor. 99. Hypoaldosteronism, hypocortisolemia, secondary and chronic hypoadrenocorticism, Addison's disease, adrenal insufficiency, pain or obesity characterized by activating mammalian apelin receptors Prevention and treatment method. 100. A method for inducing labor, which comprises activating an apelin receptor in mammals, weak labor, relaxation bleeding, uterine restoration failure, lactation deficiency, posttraumatic stress syndrome or milk stasis prevention / treatment. Method, or before and after placental delivery, Caesarean section, miscarriage or assisted abortion. 101. Cushing's disease, Cushing's syndrome, adrenocorticotropic hormone-producing pituitary tumor, CRH-producing tumor characterized by inhibiting mammalian apelin receptor.
A method for preventing and treating aldosterone-producing tumor, aldosteronism, primary cortisol resistance, congenital adrenal deficiency, Addison's disease, stress, depression, anorexia nervosa, gastroduodenal ulcer or irritable bowel syndrome. 102. Exaggerated labor pain, ankylosing uterine contraction, fetal distress, uterine rupture, cervical laceration, dysmenorrhea, preterm birth or Prad characterized by inhibiting mammalian apelin receptor.
How to prevent and treat er-Willi syndrome.