WO2002060472A1 - Remedes pour des etats d'hyponutrition - Google Patents

Remedes pour des etats d'hyponutrition Download PDF

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WO2002060472A1
WO2002060472A1 PCT/JP2002/000765 JP0200765W WO02060472A1 WO 2002060472 A1 WO2002060472 A1 WO 2002060472A1 JP 0200765 W JP0200765 W JP 0200765W WO 02060472 A1 WO02060472 A1 WO 02060472A1
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ghrelin
npy
administration
therapeutic agent
darelin
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PCT/JP2002/000765
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Japanese (ja)
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Akio Inui
Akihiro Asakawa
Toshihiro Kaga
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Chugai Seiyaku Kabushiki Kaisha
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Publication of WO2002060472A1 publication Critical patent/WO2002060472A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/25—Growth hormone-releasing factor [GH-RF], i.e. somatoliberin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/02—Nutrients, e.g. vitamins, minerals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/10—Drugs for disorders of the endocrine system of the posterior pituitary hormones, e.g. oxytocin, ADH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/10—Drugs for disorders of the endocrine system of the posterior pituitary hormones, e.g. oxytocin, ADH
    • A61P5/12—Drugs for disorders of the endocrine system of the posterior pituitary hormones, e.g. oxytocin, ADH for decreasing, blocking or antagonising the activity of the posterior pituitary hormones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid

Definitions

  • the present invention relates to a novel therapeutic agent for malnutrition symptoms. More specifically, the present invention relates to a therapeutic agent for a disease exhibiting a malnutrition symptom, which comprises darelin or a ghrelin analog as an active ingredient. The present invention also relates to a novel therapeutic agent for eating disorder or dyskinesia using a ghrelin agonist or antagonist. Background Technology ⁇
  • Weight control is based on the balance between food intake and energy expenditure, and both balances cause obesity and thinness. Since the discovery of lebutin, discovered in 1994, as a signal of adiposity (body fat mass accumulation), is implicated in the basis of weight regulation, it has been implicated in many new appetite regulation downstream of leptin The peptide was found. In particular, neuropeptides derived from the hypothalamus, which had previously only been considered as independent functions, function downstream of lebutin, and information is also exchanged densely among these neuropeptides. I understand that it is being done. Among these neuropeptides, substances that increase appetite include neuropeptide ⁇ (NPY), orexins, motilin, molanin, melanin-concentrating hormone (MCH), and agonite.
  • NPY neuropeptide ⁇
  • MCH melanin-concentrating hormone
  • Agouti-related protein is known.
  • Substances that suppress appetite include melanocyte-stimulating hormone (-MSH), corticotropin-releasing factor (CRF), cocaine- and amphetamine-regulated transcripts (CRF).
  • cocain- and amphetamine-regulated transcripts include c AR f) cholesystokinin (CCK). These peptides are involved in physiological mechanisms that control gastrointestinal motility and are thought to affect energy homeostasis.
  • NPY is a neurotransmitter consisting of 36 amino acids and is considered to be a feeding center Abundantly expressed in the hypothalamus.
  • NPY is produced in the arcuate nucleus of the hypothalamus (ARC) and is secreted mainly through the axon to the paraventricular nucleus (PVN), affecting feeding.
  • ARC arcuate nucleus of the hypothalamus
  • PVN paraventricular nucleus
  • central administration of NPY exerts a potent anorectic effect (Schwartz, MW et al., Am. J. Clin. Nutr., 69: 584, 1999), peripheral administration is not related to eating, Conversely, they tended to be suppressed. This phenomenon is seen in other PP family peptides as well.
  • NPY receptor Various physiological actions caused by NPY are performed through the NPY receptor.
  • NPY receptor five subtypes of the NPY receptor (Yl, Y2, Y4, Y5, y6) have been cloned, and its basic structure is a seven-transmembrane G protein-coupled receptor.
  • Y5 receptor was reported as a receptor closely related to the regulation of feeding, based on analysis of ligand binding specificity and feeding promoting activity. (Inui A., Trends Pharmacol Sic 20: 43-46, 1999, etc.).
  • growth hormone is a hormone secreted from the anterior pituitary gland, whose secretion is subtly controlled, stimulated by growth hormone-releasing hormone (GH RH) in the hypothalamus, and suppressed by somatostatin.
  • GHS growth hormone secretagogue
  • GHS operates via a different pathway than GHRH.
  • GHRH activates the GHRH receptor to increase the intracellular cAMP concentration
  • GHS activates a receptor different from the GHRH receptor and activates the intracellular cAMP via the intracellular IP3 system. Increase the Ca ion concentration.
  • the structure of GHS-R, a receptor on which GHS acts, was elucidated by the expression cloning method in 1996 (Howard A.D. et al, Science, 273: 974-977, 1996).
  • GHS-R is a typical G protein-coupled receptor that penetrates the cell membrane seven times, and is mainly present in the hypothalamus and pituitary gland.
  • GHS GHS-R-specific ligand
  • Darrelin is a peptide consisting of 28 amino acids, with the third serine residue n-octanoyl.
  • human darelin differs from rat darelin in two amino acid residues.
  • the structural formulas of rat and human ghrelin are shown below.
  • Chemically synthesized ghrelin has a nanomolar order of activity to increase intracellular Ca in CHO cells expressing GHSR-R and to release growth hormone in primary cultured pituitary cells. In addition, it also raises blood growth hormone in rats in vivo. Ghrelin mRNA is prominently expressed in the stomach, and dallelin is also present in the blood. In addition, GHSR is also present in the hypothalamus, heart, lung, kidney, small intestine and adipose tissue (Kojima et al., Supra). Darrelin has also been reported to have a feeding-promoting effect (Wren et al., Endocrinology, 141 (11): 4325-4328, 2000). Based on these findings, darelin is thought to be produced in the stomach and transported through the blood to the pituitary gland before exerting various effects in the brain and periphery, but its physiological role is still unknown. Not fully understood.
  • the present inventors have found that they have a remarkable appetite-promoting effect via Y and Y1 receptors, and completed the present invention.
  • the present invention provides a therapeutic agent for a disease exhibiting a malnutrition symptom, which comprises darrellin as an active ingredient.
  • the present invention further provides a method of administering a candidate substance to an animal in the presence or absence of darelin, the amount of food consumed, the amount of NPY mRNA expression, the amount of NPY bound to the Y1 receptor of NPY, and the amount of oxygen consumed.
  • a method for screening ghrelin agonists or engonist which comprises measuring gastric emptying rate or vagal activity.
  • the present invention further provides a therapeutic agent for anorexia or weight loss, comprising as an active ingredient a ghrelin agonist obtained by the above method.
  • the present invention further provides a prophylactic or therapeutic agent for obesity, which comprises, as an active ingredient, a ghrelin antagonist obtained by the above method.
  • A shows the amino acid sequences of human darelin and human motilin.
  • B shows the amino acid sequence of the human darelin receptor and the human motilin receptor. The same amino acids are indicated by an asterisk.
  • FIG. 2 shows the effect of ghrelin I CV administration on food intake.
  • FIG. 3 shows the effect of ghrelin mouse ICV administration compared to NPY, AGRP, orexin A, orexin B and MCH.
  • FIG. 4 shows NPY gene expression in the hypothalamus after ghrelin ICV administration.
  • the upper panel shows a Northern plot of hypothalamic NPY mRNA after ghrelin I CV administration.
  • the graph below shows the data from the Northern plot normalized to G3 PDH mRNA and expressed as a percentage of the control group.
  • FIG. 5 shows the effect of pretreatment with the NPY Y1 receptor antagonist (B IBO 3304) and the Y5 receptor antagonist (L152804) on dallerin-induced feeding.
  • FIG. 6 shows the effect of ghrelin I CV administration on oxygen consumption.
  • FIG. 7 shows the effect of ghrelin IP administration on food intake.
  • FIG. 8 shows the effect of darelin IP administration on hypothalamic NPY mRNA expression.
  • FIG. 9 shows the effect of darelin IP administration on gastric emptying rate.
  • FIG. 10 shows the effect of vagal nerve transection on the feeding-promoting effect of dallelin.
  • Figure 11 shows the effect of vagal transection on gastric vagal afferent activity upon administration of darelin.
  • FIG. 12 shows the results of Northern blot analysis of gastric expression of darelin mRNA in lean mice fasted for 48 hours.
  • FIG. 13 shows the results of Northern blot analysis of stomach ghrelin mRNA expression when IL-1 and lebutin were administered IP.
  • FIG. 14 shows the results of testing Darrelin mRNA expression in the stomach of obZob obese mice by Northern blot.
  • FIG. 15 shows the results of a Northern blot test of stomach expression of darelin mRNA after repeated administration of lebutin to ob / ob mice.
  • FIG. 16 shows the effect of co-administration of darelin and IL-11i8 on fasting lean mice on food intake and body weight.
  • FIG. 17 shows the effects of repeated administration of darelin on IL-1 1) 3-induced food intake and weight loss.
  • FIG. 18 shows the effect of ghrelin on weight gain in HHM / cahexia model mice transplanted with LC-6.
  • FIG. 19 shows the effect of ghrelin on increasing fat in LC-6-transplanted HHM / cahexia model mice.
  • Darrelin used as an active ingredient in the present invention is rat darelin represented by Formula 1 or human ghrelin or a ghrelin analog.
  • the darelin analogs include those in which 28 amino acids are deleted, substituted or added, as long as they have an appetite-promoting effect. Further, these derivatives, for example, peptides Derivatives in which constituent amino acids are substituted (including those in which a group such as alkylene is inserted between amino acids) and ester derivatives are also included.
  • Ghrelin or ghrelin analogs may be produced by any method, and include, for example, those isolated and purified from human and rat cells, synthetic products, semi-synthetic products, those obtained by genetic engineering techniques, and in particular, No restrictions.
  • Examples of deletions, substitutions, or additions of one or more of the eight amino acids include des-Glnl4-ghrelin, where the 14th Gin residue of darelin is deleted. It is a target. Rat des-Glnl4-ghrelin is caused by a difference in splicing of the ghrelin gene, and is present in the rat stomach in about one-fourth of ghrelin, and has the same growth hormone releasing activity as ghrelin.
  • J. Med. Chem. 2000, 43, 4370-4376 describes the minimum sequence of ghrelin required for the activation of human GHSR1a.
  • the amino acid has the third and fourth amino acids from the N-terminus (preferably the four amino acids at the N-terminus) and the third amino acid from the N-terminus (Ser ) Wherein the side chain is substituted and derivatives thereof, which have an appetite-promoting action.
  • side chain of the third amino acid from the N-terminus examples include glycerin side chains other than n-octanoyl, such as an acyl group and a substituted alkyl group (preferably having 6 to 18 carbon atoms).
  • Specific side chains include the following:
  • darelin analog having the third and fourth amino acids from the N-terminus and having the side chain of the third amino acid (Ser) from the N-terminus substituted is described in the 37th Peptide Discussion. Reported at the meeting (October 18-20, 2000): NH 2 — (CH 2 ) 4 -CO-Ser (octyl) — Ph e—L e u-NH— (CH 2 ) 2 — NH 2, and the like.
  • these ghrelin analogs can also be expected to have an appetite-promoting activity.
  • dallelin or dallelin analogue may be used in combination of two or more.
  • the therapeutic agent for a disease showing a malnutrition symptom of the present invention can be administered centrally (for example, intracerebroventricular administration, intrathecal injection) or peripherally. Preferably, it is used for peripheral administration.
  • centrally for example, intracerebroventricular administration, intrathecal injection
  • peripherally Preferably, it is used for peripheral administration.
  • NPY and other PP family peptides are not related to eating when administered peripherally, but the therapeutic agent of the present invention showed a significant appetite-enhancing effect even when administered peripherally. Therefore, the therapeutic agent of the present invention causes less pain for the patient associated with the administration and can be easily taken, which is much more advantageous than conventional appetite-regulating peptides.
  • Darrelin or a darelin analog can be made into ordinary preparations for oral administration and preparations for parenteral administration either alone or together with pharmacologically acceptable carriers and additives by known preparation techniques.
  • it is formulated into solution preparations (injections such as arterial, intravenous or subcutaneous injections, nasal drops, syrups, etc.), tablets, troches, capsules, powders, granules, ointments, suppositories, etc. be able to. It can also be used in drug delivery systems (eg, sustained release).
  • the dosage of the therapeutic agent for a disease exhibiting malnutrition of the present invention varies depending on the age, weight, symptoms, administration route, etc. of the patient, and is determined by a doctor.
  • ghrelin for intravenous administration, about 0.16 g to 100 mg / kg body weight, preferably about 0.01 mg / kg: more preferably 0.1 mg / kg body weight. lmg-: LO mg.
  • the dose is not limited to this.
  • the therapeutic agent of the present invention can be used for the treatment of a disease exhibiting malnutrition, especially from a weakness due to anorexia, cachexia or malignant disease, infectious disease and accompanying weight loss due to inflammatory disease. Effective for the selected disease.
  • it is useful as a therapeutic agent for anorexia or weight loss due to cachexia.
  • Cachexia refers to poor general condition with gradual weight loss, anemia, dry skin or edema, anorexia, etc., and the end of very many diseases such as infectious diseases, parasitosis, and malignant tumors. Appears as a symptom.
  • terms such as appetite enhancement, increased eating, and enhanced eating are used interchangeably as words having the same meaning.
  • the present invention further comprises administering a candidate substance to an animal in the presence or absence of darelin, ingesting food, NPY mRNA expression, binding of NPY to the Y1 receptor of NPY, oxygen consumption, stomach
  • a method for screening an agonist or angulinist of ghrelin or a ghrelin analog which comprises measuring the content excretion rate or the activity of a vagus nerve.
  • a specific measurement method can be, for example, the method described in the present specification, but is not limited thereto.
  • An agonist of ghrelin or a ghrelin analog obtained by the above screening method can be used as an active ingredient of the therapeutic agent for anorexia or weight loss of the present invention.
  • the ghrelin or ghrelin analog agonist obtained by the above screening method can be used as an active ingredient of the obesity preventive or therapeutic agent of the present invention.
  • pretreatment with a NPY Y1 receptor antagonist significantly inhibited dalelin-induced promotion of eating.
  • the administration of ghrelin or a ghrelin analog in the form of an angonist makes it possible not only to treat obesity but also to prevent it.
  • motilin or ghrelin or ghrelin analog is used as an agonist.
  • those agonists can be used, and as the antagonist of darelin or the ghrelin analog, motilin or the antagonist of the motilin analog can be used.
  • Motilin is a 22 amino acid residue peptide secreted by endocrine cells in the duodenum and upper jejunum (Itoh, Z., Peptides, 18: 593-608, 1997). Involved in contraction and secretion of enzymes from the stomach. Motilin has been reported to enhance GH secretion and has been reported to promote gastric motility using non-peptide motilinagost (Itoh, supra). As shown in FIG. 1A, human ghrelin and human motilin show 36% amino acid identity to each other (access numbers A593316 and P12872). In addition, as shown in FIG.
  • the human ghrelin receptor shows 50% amino acid identity as a whole with the human motilin receptor (access numbers Q922487, Q922848 and Q922). 4 3 1 9 3).
  • Tomasetto et al. Also isolated a novel peptide from the mouse stomach, which was identical to dallelin and was named a motilin-related peptide (Tomasetto C. et al., Gastroenterology, 119: 395- 405, 2000).
  • motilin or motilin analogs or agonists thereof can be used as ghrelin or ghrelin analog agonists.
  • motilin or a motilin analog antagonist can be used as the ghrelin analog antagonist.
  • NPY melanocyte stimulating hormone
  • MCH melanocyte stimulating hormone
  • CART cocaine and amphetamine monoregulated transcript
  • NPY a peptide consisting of 36 amino acids
  • CRF release factor
  • lebutin a peptide consisting of 36 amino acids
  • both the Y 1 receptor and the Y 5 receptor are considered to be NPY feeding receptors.
  • darelin administered ICV significantly stimulated food intake and reduced oxygen consumption, similar to NPY, all of which were blocked by Y1 receptor antagonists.
  • the presence of small amounts of ghrelin in the brain has been suggested by reverse transcription-polymerase chain reaction (RT-PCR) amplification and immunohistological analysis.
  • GHSR was localized in the arcuate nucleus (AC), where NPY was synthesized (Tannenbaus GS et al., Endocrinology, 139: 4420-4423). , 1998).
  • NPY mRNA expression was significantly increased by central administration of darelin.
  • the mechanism of action of ghrelin which produces a positive energy balance, may be related to the hypothalamic NP Y and Y 1 receptor systems.
  • some peptides have been shown to increase food intake when administered to the brain (eg, Elmquist JK et al., Nat Neurosci, 1: 445-450, 1998).
  • peptides that exert an appetite-enhancing effect when administered peripherally have not been reported so far.
  • the present invention has revealed that peripherally administered ghrelin stimulates food intake via NP Y and Y 1 receptors.
  • the appetite-stimulating effect of ghrelin is also mediated through vagus nerve and afferent activity It revealed that.
  • the effective dose of darelin when administered intravenously to rats is lower than that of CCK.
  • Various anorexigenic molecules, including bombesin, IL-1 ⁇ , lebutin and gastrin-releasing peptide (GRP) have been reported to increase the discharge rate of gastric vagal afferents. (See above, Schwartz, etc.).
  • the effects of darelin on vagal activity and on feeding are the opposite of these antifeedant molecules, supporting that anorectic activity acts via the vagus nerve.
  • the present invention has shown that ghrelin mRNA expression in the stomach is increased by starvation.
  • increased darrellin mRNA during starvation is at least partially responsible for the activation of hypothalamic NPY, which results in feeding.
  • the stomach is not only the source of levulin, a signal of satiation from the periphery to the hypothalamus, but also the source of darrelin, a feeding stimulus signal.
  • cytokines such as IL-1, IL-16 and tumor necrosis factor have important effects on energy balance (InuiA., Cancer Res 59: 4493-4501, 1999, etc.).
  • cachexia is known to cause poor general condition with the main symptoms such as weight loss.
  • darZin mRNA expression in the stomach is reduced by both IL-1 / 3 and leptin, and obZob mice (leptin-deficient mice that become binge eating and become obese ) Shows that it rises.
  • lebutin reduced ghrelin mRNA expression as well as energy uptake.
  • expression of the darelin gene in the stomach is linked to appetite control and nectar and plays a role in either the adaptive response to starvation or the development of obesity.
  • ghrelin reversed IL-11-induced anorexia and weight loss and improved cachexia.
  • Darrelin is known to potently stimulate growth hormone release from the pituitary gland (supra, Kojima et al.).
  • weight gain can be stimulated only by IP administration of darelin, and this peptide May contribute to the growth and control of adipose tissue mass.
  • the inventor has found that Darrelin is not a short-term diet-related olexigen (a counterpart of CCK or other diet-related satiation factors), but a long-term body weight.
  • leptin It may be a factor that regulates the activity of leptin, that is, leptin.
  • growth hormone has been used as a potential anabolic agent to treat muscle stress associated with surgical stress, sepsis, glucocorticoid administration, HIV infection and cancer. Growth hormone stimulates systemic and muscle protein synthesis, at least under certain conditions. Instead, darelin is effective in treating elderly people with decreased growth hormone secretion, decreased muscle mass, and often with anorexia.
  • the therapeutic agent for a malnutrition-related disease of the present invention which comprises ghrelin or a ghrelin analog as an active ingredient, promotes eating, reduces energy consumption, and stimulates growth hormone secretion, thereby producing a positive energy balance state. And induce weight gain.
  • mice 32-35 g were purchased from JAPAN SLC (Shizuoka, Japan). 10-11 week old obese (obZob) C57BL / 6 J mice (38-42 g) were purchased from Shionogi Co., Ltd. (Shiga, Japan). These were reared in an individually controlled environment (temperature 22 S 2C, humidity 55 ⁇ 10%, light cycle starting at 7:00 am with 12 hour light / dark cycle). Food and water were provided ad libitum unless otherwise noted. Mice were used only once in each experiment.
  • Rat Ghrelin, Rat N PY, Agouti-related protein 86-132 (AGP), Mouse Alexin A, Mouse Alexin B, and Mouse Melanin Concentrated Hormone (MCH) are purchased from Peptide Institute (Osaka, Japan) did.
  • Recombinant mouse leptin and recombinant mouse IL-11 were purchased from R & D Systems (Minneapolis, USA) and Upstate Biotechnology (New York, USA), respectively.
  • BI BO3304 was a gift from Boeringer-Ingelheim Pharma, Germany), and L152804 and J115814 were a gift from Banyu (Banyu Pharmaceutical Co., Ltd., Tokyo, Japan).
  • BI B03304 and J 115814 are NP.Y Y1 receptor antagonists, and L 152804 is a Y5 receptor antagonist.
  • each drug is diluted with artificial cerebrospinal fluid (ACSF) n1 and administered intravenously (intra-third corebroventricular: I CV), or diluted with saline 1001 and intraperitoneally (IP) was administered.
  • Control groups received ACSF or saline only.
  • Each engonist was administered at the same time as darelin. The results are shown as mean soil SE. Analysis of variance (AN OVA) was performed, and differences between groups were determined by Bonferroni's t-test. It was determined that there was a statistically significant difference in the case of P ⁇ 0.05.
  • mice are anesthetized with bentoparpital sodium (80-85 mg / kg IP) and placed in a stereotaxic apparatus (SR-6, Narishige, Tokyo, Japan) for 7 days before the experiment.
  • SR-6 stereotaxic apparatus
  • Each skull was drilled with a needle 0.9 mm lateral to the central suture, 0.9 mm posterior to the anterior crest.
  • a 24-gauge force neuron (Safelet-Cas, Nipro, Osaka, Japan) with one end inclined over a length of 3 mm was implanted in the third ventricle for ICV administration.
  • the forcenula was fixed to the skull with dental cement and capped with silicon.
  • a 27 gauge injection insert was attached to the microsyringe with PE-20 tubing.
  • vagus nerve transection was performed as follows. Mice were anesthetized with bento valpital sodium (80-8 SmgZkg IP). The midline of the abdominal wall was incised and the stomach was covered with sterile gauze moistened with warm saline. The lower esophagus was exposed and the anterior and posterior branches of the vagus nerve were cut. At the end of the operation, the abdominal wall was double sutured. Sham-operated mice also exposed the vagus nerve trunk, but did not cut it. Complete nutrition of mice with vagus nerve transection and sham-operated mice They were raised on a liquid diet (Oriental Yeast Co., Ltd. Tokyo, Japan).
  • mice were allowed to eat water ad libitum for 16 hours without food. The amount of food consumed was determined at 20 minutes, 1 hour, 2 hours, and 4 hours after the administration of ICV or IP by subtracting the amount of food left from the amount of food measured and given in advance.
  • Lean mice without food restriction were repeatedly administered IP with darelin (3 nmol / mouse), IL-1) 3 (5 picomoles / mouse) or saline for 5 days. Mice were injected daily at 7 and 19 o'clock. Food consumption and body weight were measured daily, and the fur condition was observed.
  • the total amount of the hybridized signal was determined by densitometry (Image Master ID Elite ver 3.0, Amersham Pharmacia Biotech AB, Uppsala, Sweden). The results were normalized to glyceraldenide triphosphate dehydrogenase (G3PDH) mRNA and expressed as a percentage of the control group.
  • G3PDH glyceraldenide triphosphate dehydrogenase
  • mice were fasted for 24 hours. During the fasting period, mice scheduled to receive ICV were given darelin (1 nmol / mouse) or ACSF every 12 hours and mice were killed by cervical dislocation 30 minutes after the third final dose. Mice to be administered IP were given darelin (3 nmol / mouse) or saline every 8 hours, and mice were killed by cervical dislocation 30 minutes after the last dose. Immediately The brain block was excised, frozen on dry ice, and stored at 180 ° C until a Northern blot was prepared.
  • mice were fasted for 48 hours. During the fasting period, fasted mice were given IP 1/3 (5 picomoles / mouse), lebutin (3 nanomoles / mouse), or saline every 12 hours, and 30 minutes after the final dose of 5th dose The mice were killed by cervical dislocation. In non-fasted ob / ob mice, leptin (3 nmole Z mice) or saline was administered repeatedly for 7 days. Mice were injected daily at 7 and 19 o'clock and 30 minutes after the last dose mice were killed by cervical dislocation. Immediately the stomach was excised, frozen on dry ice, and stored at -80 "C until Northern blots were prepared.
  • Oxygen consumption 2 2 0 2 Bruno C_ ⁇ 2 metabolic measurement system (Model MK-5000, Muromachikikai, Tokyo, Japan) was measured by using the (supra, Ueno N. et al.).
  • the chamber volume was 560 ml, and the air flow rate to the chamber was 500 ml Z min. Samples were taken every 3 minutes and standard gas samples were taken every 30 minutes. Mice were placed in the light cycle with no food, no water and no restraint in the chamber, and ghrelin (at 10 o'clock) in the presence or absence of BIBO334 (5 nanomolar mice). (3-1 nmole mouse) was administered ICV, and the oxygen consumption was measured 2 hours later.
  • the effect of ghrelin on vagal activity was measured by comparing the average number of impulses every 5 seconds over 50 seconds before and after injection. The results were expressed as mean soil SE. ANOVA and Scheffe tests were performed to evaluate differences between groups. The case of P ⁇ 0.05 was judged to be statistically significant.
  • Example 3 Effect of Darrelin I CV Administration on NPY Gene Expression
  • FIG. 1 shows a Northern plot of hypothalamic NPY mRNA after ghrelin I CV administration.
  • the graph below shows the Northern blot data normalized to glyceraldenide triphosphate dehydrogenase (G3PDH) mRNA and expressed as a percentage of the control group.
  • G3PDH glyceraldenide triphosphate dehydrogenase
  • IP-administered ghrelin exerts a similar effect of increasing food intake.
  • IP administration of darelin significantly increased feeding at 3 nmol / mouse ( Figure 7).
  • the accumulated food intake at 24 hours after IP administration of 3 nmoles / mouse darrellin was even higher (6.68 ⁇ 0.16 g vs. 6.10 ⁇ 0.17 g (control): P ⁇ 0.05) ).
  • This anorectic activity was blocked by ICV administration of the Y1 receptor antagonist gonivist BIB03304, but not by the Y5 receptor antagonist gonist L152804.
  • FIG. 9 shows the gastric emptying rate at 1 hour and 2 hours after darelin (0.3-1 nmol / mouse) was administered by ICV.
  • Ghrelin significantly increased gastric emptying rate 1 hour post-ICV or IP (30.16 ⁇ 3.70% (3 nmol) vs. 20.34 ⁇ 2.27% (control)) : P ⁇ 0. 05).
  • ghrelin like motilin, enhances gastric motility.
  • Example 8 Effect of Vagal Nerve Surgery on Dalelin's Feeding-Promoting Effect, NPY mRNA Expression, and Gastric Vagal Nerve Afferent Activity

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Abstract

L'invention concerne des remèdes pour des maladies relatives à des états d'hyponutrition, tels que l'inappétence, l'émaciation, ou des maladies malignes, et la prostration, provoquées par une perte de poids, ainsi qu'une infection ou des maladies inflammatoires. Ces remèdes contiennent, en tant que principe actif, de la ghréline ou ses analogues.
PCT/JP2002/000765 2001-01-31 2002-01-31 Remedes pour des etats d'hyponutrition WO2002060472A1 (fr)

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Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004004772A1 (fr) * 2002-07-05 2004-01-15 Chugai Seiyaku Kabushiki Kaisha Traitement antidiabetique
WO2004096260A1 (fr) * 2003-04-30 2004-11-11 Kenji Kangawa Prophylactiques ou remedes pour l'hepatopathie
WO2005110463A1 (fr) * 2004-05-11 2005-11-24 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services, National Institutes Of Health Procedes pour inhiber l'expression de cytokine pro-inflammatoire au moyen de ghreline
EP1694355A1 (fr) * 2003-10-24 2006-08-30 THE ACADEMIC HOSPITAL ROTTERDAM acting under the name Erasmus MC Utilisation de compositions de ghreline et de ghreline non acylee dans des etats pathologiques lies a l'insuline
CN1321131C (zh) * 2003-12-26 2007-06-13 李宁 猪Ghrelin衍生物及其编码基因与应用
US7666833B2 (en) 2001-12-18 2010-02-23 Alizé Pharma SAS Pharmaceutical compositions comprising unacylated ghrelin and therapeutical uses thereof
US7803768B2 (en) 2005-02-23 2010-09-28 Kyoto University Method for treatment of hyperglycemia in a subject in need thereof
JP2011511835A (ja) * 2008-02-13 2011-04-14 シージーアイ ファーマシューティカルズ，インコーポレーテッド ６−アリール−イミダゾ［１，２−ａ］ピラジン誘導体、その製造方法、及びその使用方法
WO2011087102A1 (fr) * 2010-01-15 2011-07-21 国立大学法人宮崎大学 Agent thérapeutique pour encourager la récupération d'un animal sous traitement médical
CN102552754A (zh) * 2012-02-23 2012-07-11 青岛绿曼生物工程有限公司 促进肉禽生长育肥的复方蜂胶组合物及其制备方法
US8222217B2 (en) 2007-05-31 2012-07-17 Alize Pharma Sas Unacylated ghrelin as therapeutic agent in the treatment of metabolic disorders
US8318664B2 (en) 2007-05-31 2012-11-27 Alize Pharma Sas Unacylated ghrelin fragments as therapeutic agent in the treatment of obesity
US8476408B2 (en) 2008-06-13 2013-07-02 Alize Pharma Sas Unacylated ghrelin and analogs as therapeutic agents for vascular remodeling in diabetic patients and treatment of cardiovascular disease
US9550821B2 (en) 2011-12-15 2017-01-24 Alize Pharma Sas Modulation of ghrelin levels and ghrelin/unacylated ghrelin ratio using unacylated ghrelin
JP2021107358A (ja) * 2019-12-27 2021-07-29 クラシエ製薬株式会社 グレリン受容体の活性化剤

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6456015B2 (ja) * 2013-07-19 2019-01-23 国立大学法人三重大学 閉塞性動脈硬化症モデル動物、るい痩研究用モデル動物、及び全身性アミロイドーシスモデル動物としての非ヒト哺乳動物

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001056592A1 (fr) * 2000-02-01 2001-08-09 Novo Nordisk A/S Utilisation de composes pour la regulation de l'absorption de nourriture
WO2001087335A2 (fr) * 2000-05-17 2001-11-22 Eli Lilly And Company Procede d'inhibition selective de la ghreline

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001056592A1 (fr) * 2000-02-01 2001-08-09 Novo Nordisk A/S Utilisation de composes pour la regulation de l'absorption de nourriture
WO2001087335A2 (fr) * 2000-05-17 2001-11-22 Eli Lilly And Company Procede d'inhibition selective de la ghreline

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
NAKAZATO MASAMITSU ET AL.: "A role for ghrelin in the central regulation of feeding", NATURE, vol. 409, 11 January 2001 (2001-01-11), pages 194 - 198, XP002901678 *
TSCHOP MATTHIAS ET AL.: "Ghrelin induces adiposity in rodents", NATURE, vol. 407, 2000, pages 908 - 913, XP002951587 *
WREN A.M. ET AL.: "The novel hypothalamic peptide ghrelin stimulates food intake and growth hormone secretion", ENDOCRINOLOGY, vol. 141, no. 11, 2000, pages 4325 - 4328, XP002951588 *

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US7666833B2 (en) 2001-12-18 2010-02-23 Alizé Pharma SAS Pharmaceutical compositions comprising unacylated ghrelin and therapeutical uses thereof
WO2004004772A1 (fr) * 2002-07-05 2004-01-15 Chugai Seiyaku Kabushiki Kaisha Traitement antidiabetique
WO2004096260A1 (fr) * 2003-04-30 2004-11-11 Kenji Kangawa Prophylactiques ou remedes pour l'hepatopathie
US10653637B2 (en) 2003-04-30 2020-05-19 Kenji Kangawa Preventives or remedies for hepatopathy
CN1780635B (zh) * 2003-04-30 2011-04-13 寒川贤治 肝病的预防或治疗剂
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EP1694355A1 (fr) * 2003-10-24 2006-08-30 THE ACADEMIC HOSPITAL ROTTERDAM acting under the name Erasmus MC Utilisation de compositions de ghreline et de ghreline non acylee dans des etats pathologiques lies a l'insuline
EP1694355A4 (fr) * 2003-10-24 2009-07-29 Alize Pharma Sas Utilisation de compositions de ghreline et de ghreline non acylee dans des etats pathologiques lies a l'insuline
CN1321131C (zh) * 2003-12-26 2007-06-13 李宁 猪Ghrelin衍生物及其编码基因与应用
JP2007537276A (ja) * 2004-05-11 2007-12-20 ザ ガバメント オブ ザ ユナイテッド ステイツ オブ アメリカ， アズ リプレゼンティッド バイ ザ セクレタリー， デパートメント オブ ヘルス アンド ヒューマン サービシーズ， ナショナル イ グレリンを用いて炎症誘発性のサイトカイン発現を阻害する方法
WO2005110463A1 (fr) * 2004-05-11 2005-11-24 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services, National Institutes Of Health Procedes pour inhiber l'expression de cytokine pro-inflammatoire au moyen de ghreline
US7803768B2 (en) 2005-02-23 2010-09-28 Kyoto University Method for treatment of hyperglycemia in a subject in need thereof
US8222217B2 (en) 2007-05-31 2012-07-17 Alize Pharma Sas Unacylated ghrelin as therapeutic agent in the treatment of metabolic disorders
US8318664B2 (en) 2007-05-31 2012-11-27 Alize Pharma Sas Unacylated ghrelin fragments as therapeutic agent in the treatment of obesity
JP2011511835A (ja) * 2008-02-13 2011-04-14 シージーアイ ファーマシューティカルズ，インコーポレーテッド ６−アリール−イミダゾ［１，２−ａ］ピラジン誘導体、その製造方法、及びその使用方法
US8476408B2 (en) 2008-06-13 2013-07-02 Alize Pharma Sas Unacylated ghrelin and analogs as therapeutic agents for vascular remodeling in diabetic patients and treatment of cardiovascular disease
WO2011087102A1 (fr) * 2010-01-15 2011-07-21 国立大学法人宮崎大学 Agent thérapeutique pour encourager la récupération d'un animal sous traitement médical
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US9550821B2 (en) 2011-12-15 2017-01-24 Alize Pharma Sas Modulation of ghrelin levels and ghrelin/unacylated ghrelin ratio using unacylated ghrelin
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JP2021107358A (ja) * 2019-12-27 2021-07-29 クラシエ製薬株式会社 グレリン受容体の活性化剤
JP7430442B2 (ja) 2019-12-27 2024-02-13 クラシエ株式会社 グレリン受容体の活性化剤

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